PMID- 15745892 TI - Magnesium sulfate for prevention of postoperative atrial fibrillation in patients undergoing coronary artery bypass grafting. AB - PURPOSE: The effects of i.v. magnesium sulfate on the frequency of postoperative atrial fibrillation (AF) in patients undergoing coronary artery bypass grafting (CABG) and on the frequency of AF at hospital discharge were studied. The effect of postoperative AF on hospital length of stay (LOS) was also assessed. METHODS: A retrospective chart review was performed for all patients who underwent CABG surgery by a single surgeon during 2000-2001 at a community medical center. Patients were eligible for inclusion if they had first-time CABG surgery. Patients who underwent CABG surgery in 2000 did not receive magnesium sulfate and served as controls for the study. Patients were included in the magnesium group if they received 2 g of i.v. magnesium sulfate intraoperatively and 2 g every 12 hours postoperatively for at least two consecutive days. RESULTS: A total of 262 patients underwent CABG during the study period, and 28 were excluded from the study. Of the remaining 234 patients, 99 were in the magnesium group, and 135 were in the control group. No significant differences were found between the study groups in recorded demographic characteristics. Postoperative AF occurred significantly less frequently in the magnesium group (p = 0.038). There was no significant difference between treatment groups in the number of patients discharged in AF (p = 0.307). Among all patients, those with AF were significantly more likely to have a prolonged LOS (p = 0.036). CONCLUSION: CABG patients who received intraoperative and postoperative i.v. magnesium sulfate had a significantly lower rate of AF compared with patients who did not receive the drug. The number of patients discharged with AF was not affected by magnesium administration. AF was associated with a higher likelihood of prolonged postoperative hospitalization. PMID- 15745893 TI - Epoetin alfa protocol and multidisciplinary blood-conservation program for critically ill patients. AB - PURPOSE: An evidence-based epoetin alfa protocol and a multidisciplinary blood conservation program were implemented in a medical intensive care unit (MICU) and surgical intensive care unit (SICU). SUMMARY: Baseline data were collected to evaluate the use of epoetin alfa and red blood cell (RBC) transfusions in our MICU and SICU. An evidence-based protocol for epoetin alfa use and a multidisciplinary blood-conservation program were designed, approved, and implemented. Preprotocol patients consisted of a convenience sample of 18 patients receiving epoetin alfa for various indications who were admitted to our MICU and SICU from January 1 to December 31, 2002. The postprotocol sample consisted of 40 patients who received epoetin alfa for the treatment of anemia due to critical illness who were admitted to the MICU and SICU from March 25 to May 23, 2003. Postprotocol data were collected and compared with baseline data. All patients seen in the MICU and SICU, during the postprotocol period, regardless of whether they were receiving epoetin alfa, were included in the multidisciplinary blood-conservation program. Postprotocol data showed statistically significant improvements in epoetin alfa dosing and monitoring and in the use of adjunctive therapy. Pharmacist-initiated blood-conservation strategies resulted in several blood-draw reductions and discontinuations. Statistically significant reductions in the number of RBC units transfused per patient and per intensive care unit (ICU) day were also observed. CONCLUSION: An epoetin alfa protocol and a multidisciplinary blood-conservation program contributed to rational prescribing of epoetin alfa and to a reduction in the number of RBC units transfused per patient and per ICU day. PMID- 15745894 TI - Telepharmacy in a health maintenance organization. AB - PURPOSE: A telepharmacy service in a health maintenance organization is described. SUMMARY: Kaiser Permanente Colorado Region's clinical pharmacy call center (CPCC) was established in 1996 after an audit showed that the third most common type of call to the regional call center involved questions about drug therapy. The service was developed through collaboration among the pharmacy department, the medical group, and the health plan to care for patients with medication-related questions. CPCC pharmacists work closely with the physicians, nurses, and service associates of the regional call center; primary care physicians; and other pharmacy department members. The CPCC team answers approximately 1200 telephone calls daily, including calls from new members regarding their transition into the health plan, calls about medications for patients recently discharged from outside hospitals, questions about medication related news releases, and general drug therapy questions. CPCC pharmacists collaborate with physicians to care for patients with allergic rhinitis and to complete projects that improve the quality and cost-effectiveness of drug therapy regimens. Many of CPCC's interactions are entered into an electronic medical record. CPCC has helped over 40,000 new members with their transition into the health plan since January 2000, and significant cost savings have resulted. CONCLUSION: CPCC has used telephonic, electronic, and other means of communication in an effort to reduce costs and improve the quality of care. PMID- 15745895 TI - Assessing adherence to statin therapy using patient report, pill count, and an electronic monitoring device. PMID- 15745896 TI - Effect of a computerized prescriber-order-entry system on reported medication errors. PMID- 15745897 TI - Seizures associated with memantine use. PMID- 15745898 TI - Contract price integrity for pharmaceuticals. PMID- 15745899 TI - Expanding pharmacy's reach across the care continuum. PMID- 15745900 TI - Student's death sparks concerns about compounded preparations. PMID- 15745901 TI - Clinical pharmacy demonstration program gets a review. PMID- 15745902 TI - FDA advisers vote against nonprescription lovastatin. PMID- 15745903 TI - Increasing hospital pharmacy revenue. PMID- 15745904 TI - Patients' response to revised practices for treating chemotherapy-induced nausea and vomiting. PMID- 15745905 TI - Compliance of a VA pharmacy with documentation requirements for human research. PMID- 15745906 TI - Linezolid pharmacokinetics in an obese patient. PMID- 15745907 TI - Antihyperglycemics during the perioperative period. PMID- 15745908 TI - New practitioners' role in achieving the ASHP Health-System Pharmacy 2015 Initiative. PMID- 15745909 TI - Demand clean vials. PMID- 15745910 TI - Surface contamination of chemotherapy drug vials and evaluation of new vial cleaning techniques: results of three studies. AB - PURPOSE: The results of three studies that describe the external contamination of chemotherapy drug vials are presented. New techniques for the improved decontamination of vials containing cisplatin are also described. SUMMARY: Study 1 evaluated the external contamination of drug vials with cyclophosphamide and ifosfamide in a pharmacy setting. Widespread contamination of the outside of drug vials was found with each drug. Study 2 evaluated the surface contamination of drug vials with cyclophosphamide and fluorouracil in three pharmacies. Sporadic contamination with fluorouracil was detected, while cyclophosphamide was found on most vials. In study 3, investigators compared the decontamination abilities of a standard decontamination procedure at the manufacturer level with an improved decontamination procedure and the use of sleeves to further decrease contamination. Though the methods of each study reported herein differed, the outcomes were similar. All chemotherapy drug vials studied demonstrated levels of contamination with the drug well above the limit of detection. Improved decontamination procedures, combined with the use of protective sleeves, reduced the level of platinum contamination by 90%, suggesting that standard decontamination procedures should be reconsidered. CONCLUSION: The results of these studies are consistent with several others that have reported contamination of the outside surface of drug vials for a number of chemotherapy drugs. Contamination can be reduced by using decontamination equipment and protective sleeves during the manufacturing process. PMID- 15745911 TI - Formulation and efficacy of triamcinolone acetonide mouthwash for treating oral lichen planus. AB - PURPOSE: The stability of a triamcinolone acetonide mouthwash and its efficacy in treating oral lichen planus are described. METHODS: The solubility of triamcinolone acetonide in ethanol, propylene glycol, and glycerin was determined by shaking and equilibrating an excess of triamcinolone acetonide with the solvents for 72 hours. All three solvents were used in formulating a mouthwash. A stock solution of triamcinolone acetonide standard was prepared in ethanol and diluted to yield concentrations of 2, 4, 8, 12, and 16 microg/mL. Analytical sample solutions were prepared by pipetting 0.1 mL of triamcinolone acetonide mouthwash into 10-mL volumetric flasks and diluting to volume with the mobile phase. Accelerated stability studies were conducted by storing the samples in 60 mL amber glass bottles at 45, 60, 70, and 80 degrees C and 75% relative humidity until the triamcinolone concentration decreased markedly. Efficacy was tested by 20 subjects with a clinical diagnosis of and histologically confirmed symptomatic oral lichen planus who were randomized to use the mouthwash (n = 11) or the commercially available triamcinolone acetonide paste (n = 9). RESULTS: The mouthwash had a satisfactory shelf life and was well accepted by patients. Ten of 11 patients treated with the mouthwash for four weeks reported a positive response, and a complete response in signs and symptoms occurred in 4 and 5 of 11 patients, respectively. No significant difference in clinical improvement was observed between groups. CONCLUSION: A triamcinolone acetonide mouthwash had a satisfactory shelf life and was well accepted by patients. It did not have a significantly different therapeutic efficacy from the commercial paste dosage form in the treatment of oral lichen planus. PMID- 15745912 TI - Update on the pharmacist shortage: national and state data through 2003. AB - PURPOSE: Aggregate Demand Index (ADI) survey results were used to describe the severity of the pharmacist shortage at the national and state levels and by practice site and impact on the U.S. population. METHODS: Time-series analysis was used to characterize national and state trends in the ADI from September 1999 through September 2003. The time trends for the distribution of ratings and the demand index by practice site were also examined. Historical data about retail prescriptions filled and related growth rates were compiled and compared. ADI survey results were also compared with data from other surveys. RESULTS: Over time, ADI data demonstrated a continuing national pharmacist shortage, as the ability to fill pharmacist vacancies was rated at least moderately difficult. A very slight downward trend in severity (slope = -0.008) was observed. Other survey series had similar findings. States with the most severe shortage levels tended to have large populations, while those with the lowest levels tended to have smaller populations. More states improved than worsened the severity of their shortage, with 30 states maintaining the same ADI rating. Although there was a high correlation between the retail prescription growth rate and the ADI (r = 0.84), there was a much greater decrease in prescription growth (73%) than ADI levels (6.5%). CONCLUSION: There was a sustained unmet demand for pharmacists throughout the United States from September 1999 through September 2003. More states moved toward having an adequate supply of pharmacists than toward having a more severe shortage of pharmacists, but the national ADI suggests that the system remains stressed. PMID- 15745913 TI - Hospital rules-based system: the next generation of medical informatics for patient safety. AB - PURPOSE: The hospital rules-based system (HRBS) and its subsystems at a major medical center are described. SUMMARY: The HRBS was implemented at the Mayo Clinic to rapidly identify and communicate crucial information to the clinician in order to optimize patient care. The system also enhances workload efficiency and improves documentation and communication. The system is used by the infectious-diseases division, pharmacy services, nutritional support services, infection control, and the nursing department. The six HRBS subsystems are Web based programs that share a common structural design and integrate computerized information from multiple institutional databases. The integrated data are presented in a user-friendly format that improves the efficiency of data retrieval. Information, such as monitoring notes and intervention information, can be entered for specific patients. The subsystems use rules designed to detect suboptimal therapy or monitoring and identify opportunities for cost savings in a timely manner. CONCLUSION: The HRBS enhances the identification of drug-related problems while optimizing patient care and improving communication and efficiency at a major medical center. PMID- 15745914 TI - Stability of cidofovir in extemporaneously prepared syringes. PMID- 15745915 TI - Initiating basal insulin therapy in patients with type 2 diabetes mellitus. PMID- 15745916 TI - 2004 ASHP National Residency Preceptors Conference explores generational differences, leadership development. PMID- 15745917 TI - New economics of the pharmaceutical supply chain. PMID- 15745918 TI - Changing dynamics in the pharmaceutical supply chain: a GPO perspective. PMID- 15745919 TI - Implementation of an i.v. medication safety system. PMID- 15745920 TI - Sleep attacks in patients receiving dopamine-receptor agonists. PMID- 15745921 TI - Consensus statement on the management of the GH-treated adolescent in the transition to adult care. AB - The European Society for Paediatric Endocrinology held a consensus workshop in Manchester, UK in December 2003 to discuss issues relating to the care of GH treated patients in the transition from paediatric to adult life. Clinicians experienced in the care of paediatric and adult patients on GH treatment, from a wide range of countries, as well as medical representatives from the pharmaceutical manufacturers of GH participated. PMID- 15745922 TI - Increase of interferon-gamma inducible alpha chemokine CXCL10 but not beta chemokine CCL2 serum levels in chronic autoimmune thyroiditis. AB - OBJECTIVE: To measure serum levels of CXCL10 and CCL2 prototype chemokines of the two major subclass (CXC and CC) in patients with newly diagnosed chronic autoimmune thyroiditis (AT), and relate the findings to the clinical phenotype. DESIGN AND METHODS: Serum CXCL10 and CCL2 were assayed in 70 consecutive patients with newly diagnosed chronic AT, in sex- and age-matched healthy volunteers (n = 37) and in 20 patients with non-toxic multinodular goiter, extracted from a random sample of the general population from the same geographic area. RESULTS: CXCL10 serum levels were significantly higher in patients with thyroiditis than in controls or multinodular goiter patients, while comparable CCL2 levels were found between groups. CXCL10 levels were significantly increased in hypothyroid patients and in those with an hypoechoic pattern (P = 0.0004 and P = 0.0001, respectively) while serum CCL2 levels were significantly increased in patients older than 50 years and in those with hypothyroidism (P = 0.0001 and P = 0.03, respectively). No correlation between CXCL10 and CCL2 serum levels could be demonstrated. CXCL10 and CCL2 were studied separately in relation to clinical features of AT patients. Two separate multiple linear regression models for CXCL10 and CCL2 were performed, including age, thyroid volume, thyroid stimulating hormone (TSH), FT4, anti-thyroid peroxidase (AbTPO), hypoechoic pattern, and the presence of hypervascularity, demonstrating that ln of serum CXCL10 levels was associated with TSH independently of other possible confounders levels [regression coefficient (R.C.) 0.143 confidence interval (C.I.) (0.042 0.245); P = 0.0059], while serum CCL2 were significantly associated only with age [R.C. 5.412 C.I. (3.838-6.986); P < 0.0001]. CONCLUSION: Our results, obtained in a large cohort of newly diagnosed AT patients demonstrate increased CXCL10 especially in hypothyroid patients with a more aggressive disorder, and normal CCL2 serum levels in AT. PMID- 15745923 TI - L-tri-iodothyronine is a major determinant of resting energy expenditure in underweight patients with anorexia nervosa and during weight gain. AB - OBJECTIVE: We aimed to define the effect of L-3,5,3'-tri-iodothyronine (T(3)) on metabolic adaptation in underweight patients with anorexia nervosa (AN) as well as during weight gain. METHODS: This involved clinical investigation of 28 underweight patients with AN, who were compared with 49 normal-weight controls. A subgroup of 17 patients was followed during weight gain. Resting energy expenditure was measured by indirect calorimetry. Body composition was measured by anthropometry as well as bioelectrical impedance analysis. Energy intake (EI) was assessed by a 3-day dietary record. Plasma concentrations of thyroid hormones (thyroxine (T(4)), T(3) and thyrotropin (TSH)) were analyzed by enzyme immunoassays. RESULTS: When compared with normal-weight women, underweight patients with AN had reduced fat mass (FM) (-71.3%), fat-free mass (FFM) ( 13.1%), resting energy expenditure (REE) (-21.8%), T(3)- (-33.4%) and T(4) concentrations (-19.8%) at unchanged TSH. REE remained reduced after adjustment for FFM (-24.6%). T(3) showed a close association with REE. This association remained after adjustment of REE for FFM. Treatment of underweight AN patients resulted in a mean weight gain of 8.3 kg. This was mainly explained by an increase in FM with small or no changes in FFM. REE and T(3) also increased (+9.3% and +33.3% respectively) at unchanged TSH and T(4). There was a highly significant association between weight gain-induced changes in T(3) and changes in adjusted REE (r = 0.78, P < 0.001, based on Pearson's correlation). An increase in plasma T(3) concentrations of 1.8 pmol/l could explain an increase in REE of 0.6 MJ/day (that is, a 32% increase in T(3) was associated with a 13% increase in REE). CONCLUSIONS: Our data provide evidence that the low T(3) concentrations add to metabolic adaptation in underweight patients with AN. During weight gain, increases in T(3) are associated with increases in REE, which is independent of FFM. Both results are evidence for a physiologic role of T(3) in modulation of energy expenditure in humans. PMID- 15745924 TI - Thyroid and adrenal axis in major depression: a controlled study in outpatients. AB - OBJECTIVE: Major depressive disorder has been associated with changes in the hypothalamus-pituitary-thyroid (HPT) axis and with hypercortisolism. However, the changes reported have been at variance, probably related to in- or outpatient status, the use of antidepressant medication and the heterogeneity of depression. We therefore conducted a controlled study in unipolar depressed outpatients who had been free of antidepressants for at least 3 months. DESIGN: We assessed endocrine parameters in 113 depressed outpatients and in 113 sex- and age-matched controls. METHODS: Patients were included if they had a major depression according to a Structural Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders (DSM), fourth edition (SCID-IV) and if they had a 17 item Hamilton rating scale for depression (HRSD) score of > or =16. Endocrine parameters contained serum concentrations of TSH, (free) thyroxine, tri iodothyronine, cortisol, thyroid peroxidase (TPO) antibody titre and 24-h urinary excretion of cortisol. RESULTS: The serum concentration of TSH was slightly higher in depressed patients as compared with controls (P < 0.001), independent of the presence of subclinical hypothyroidism and/or TPO antibodies (n = 28). All other HPT axis parameters were similar in both groups. The 24-h urinary cortisol excretion was similar in patients and controls. In atypical depression, serum cortisol was lower than in non-atypical depression (P = 0.01). Patients with neither melancholic depression nor severe depression (HRSD > or =23) had altered endocrine parameters. Finally, serum TSH values could not be related to cortisol values. CONCLUSION: When compared with matched control subjects, outpatients with major depression had slightly higher serum TSH, while urinary cortisol levels were similar. Furthermore, we observed lower serum cortisol in atypical depression than in non-atypical depression. PMID- 15745925 TI - Mutation screening of the thyroid peroxidase gene in a cohort of 55 Portuguese patients with congenital hypothyroidism. AB - OBJECTIVE: Defects in the human thyroid peroxidase (TPO) gene are reported to be one of the causes of congenital hypothyroidism (CH) due to a total iodide organification defect. The aim of the present study was to determine the nature and frequency of TPO gene mutations in patients with CH, characterised by elevated TSH levels and orthotopic thyroid gland, identified in the Portuguese National Neonatal Screening Programme. SUBJECTS AND METHODS: The sample comprised 55 patients, from 53 unrelated families, with follow-up in the endocrinology clinics of the treatment centres of Porto and Lisbon. Mutation screening in the TPO gene (exons 1-17) was performed by single-strand conformational analysis followed by sequencing of fragments with abnormal migration patterns. RESULTS: Eight different mutations were detected in 13 patients (seven homozygotes and six compound heterozygotes). Novel mutations included three missense mutations, namely 391T > C (S131P), 1274A > G (N425S) and 2512T > A (C838S), as well as the predictable splice mutation 2748G > A (Q916Q/spl?). The undocumented polymorphism 180-47A > C was also detected. CONCLUSION: The results are in accordance with previous observations confirming the genetic heterogeneity of TPO defects. The proportion of patients in which the aetiology was determined justifies the implementation of this molecular testing in our CH patients with dyshormonogenesis. PMID- 15745926 TI - Chronic glucocorticoid treatment alters spontaneous pulsatile parathyroid hormone secretory dynamics in human subjects. AB - OBJECTIVE: Spontaneous parathyroid hormone (PTH) secretory dynamics include tonic and pulsatile components. It is not known how glucocorticoids might alter these secretory dynamics. DESIGN: The aim of our study was to evaluate spontaneous fluctuations in serum PTH levels in six adult male patients (aged 31-64 years) receiving chronic (>6 months) therapy with glucocorticoids (daily dosage >7.5 mg of prednisone or dose equivalent of other corticosteroid) as compared with a control group of 10 age- and sex-matched normal subjects. METHODS: Peripheral venous blood sampling was performed every 3 min for 6 h from 0900 to 1500 h. Plasma PTH release profiles were subjected to deconvolution analysis, a method that resolves measured hormone concentrations into secretion and clearance components, and to an approximate entropy (ApEn) estimate, that in turn provides an integrated measure of the serial regularity or orderliness of the release process. RESULTS: In the glucocorticoid-treated group, the PTH tonic secretory rate was reduced (4.3+/-0.74 vs 8.8+/-1.4 pg/ml per min in controls, P = 0.017). There was, however, an increase in the fractional pulsatile PTH secretion (42+/ 8.2 vs 18.3+/-3.9 pg/ml per min, P = 0.006) in glucocorticoid-treated vs normal subjects. Mean overall PTH concentration, as well as mean integrated area, was similar among normal and glucocorticoid-treated subjects. CONCLUSIONS: These results demonstrate, for the first time, that chronic glucocorticoid treatment induces a redistribution of spontaneous PTH secretory dynamics by reducing the amount released in tonic fashion and increasing the amount released as pulses. PMID- 15745927 TI - Serum cytokine levels in GH-deficient children during substitutive GH therapy. AB - OBJECTIVE: The aim of the present study was to investigate the effect of exogenously administered human GH (hGH) on serum levels of interleukin (IL)-4, IL 6, IL-12 and tumour necrosis factor (TNF)-alpha in GH-deficient (GHD) children. DESIGN AND METHODS: We evaluated 13 short prepubertal GHD children, aged between 2 and 13 years, and 13 age-matched healthy subjects as controls. Circulating cytokine values were evaluated in basal conditions in all children, and 6 and 24 h following the 1st hGH injection (0.23 mg/kg per week), and then after 3 months of hGH treatment in GHD patients. Serum levels of IL-4, IL-6, IL-12 and TNF-alpha were measured by commercially available ELISAs. RESULTS: No significant differences were found between controls and GHD children in basal values of serum IL-4, IL-6, IL-12 and TNF-alpha (P > 0.05 by Mann-Whitney U test). Analysis of cytokine levels during hGH treatment showed significant changes over time in TNF alpha and IL-6 levels (P = 0.0014 and P = 0.00 024 respectively), with the more pronounced effect observed at 6 h following the first administration of hGH (i.e. increase in IL-6 (Wilcoxon matched pairs test, P = 0.0015) and TNF-alpha levels (P = 0.0015)). No significant changes over time were observed in IL-4 and IL-12 serum levels. CONCLUSIONS: In vivo release of the pro-inflammatory cytokines IL-6 and TNF-alpha can be affected by hGH treatment in GHD children, suggesting a direct effect of GH on the immune function. PMID- 15745928 TI - Effect of recombinant human GH on circulating granulocyte colony-stimulating factor and neutrophils in patients with adult GH deficiency. AB - BACKGROUND: We previously reported that short-term continuous subcutaneous infusion (CSI) of recombinant human growth hormone (rhGH) increased plasma erythropoietin levels and hemoglobin concentrations in patients with adult GH deficiency. In the present study, we investigated the effect of rhGH on plasma granulocyte colony-stimulating factor (G-CSF) levels and neutrophil counts in patients with adult GH deficiency. METHODS: rhGH was administrated for 1 year in six patients with adult GH deficiency (age range, 24-69 years; mean +/- S.E.M., 51.7 +/- 5.8 years; two males and four females) by means of CSI at a rate of 0.25 U/kg per week. Blood samples were obtained in the morning after overnight fasting every month before and after the start of rhGH administration. Plasma GH, insulin like growth factor I (IGF-I) and G-CSF levels, and neutrophil counts, were measured. RESULTS: Mean ( +/- S.E.M.) plasma GH levels increased from 0.26 +/- 0.14 to 2.28 +/- 0.20 microg/l 1 month after the start of rhGH administration. An increase of the plasma GH levels was accompanied by an increase in the plasma IGF I levels from 64.7 +/- 8.5 to 293.3 +/- 80.6 microg/l. Plasma G-CSF levels increased at 2, 3, 8, 9 and 10 months after the start of rhGH administration compared with 28.6 +/- 11.0 ng/l at time 0. The neutrophil counts increased at 2, 3, 7, 8, 9, 11 and 12 months after the start of rhGH administration compared with 2822 +/- 377 neutrophils/microl at time 0. CONCLUSION: rhGH administration increased plasma G-CSF levels and neutrophil counts. GH and/or IGF-I might stimulate neutrophil production and/or release via G-CSF. PMID- 15745929 TI - The 'bio-assay' quality of life might be a better marker of disease activity in acromegalic patients than serum total IGF-I concentrations. AB - OBJECTIVES: To investigate the quality of life (QoL) in acromegalic patients in relation to biochemical parameters. DESIGN AND METHODS: Single-center, open label study in 14 acromegalic patients (eight woman and six men, age 33-77 years), with normal serum IGF-I levels during long-term treatment with monthly injections of 20 mg of long-acting octreotide. We investigated which biochemical parameter might reflect optimal QoL, using the SF-36 questionnaire. RESULTS: We observed that six patients had a low QoL score at baseline in the same range as observed in cancer patients. The other eight patients had a normal QoL. GH, IGF-I nor free IGF-I could discriminate these two subgroups at baseline. After skipping one monthly injection, all six subjects with the low QoL escaped in their free IGF-I concentrations. Also total IGF-I concentrations escaped in four of these six. In the subjects with normal QoL, free IGF-I levels remained normal in all, while total IGF-I levels only escaped in one. CONCLUSIONS: This study tells us that the currently used biochemical criteria for disease control in acromegaly might be sufficient in assessing long-term mortality and morbidity, but they are insufficient in addressing the most important parameter from the patient's perspective--QoL. PMID- 15745930 TI - Relationship between serum adiponectin concentration, pulse wave velocity and nonalcoholic fatty liver disease. AB - OBJECTIVES: We aimed to investigate the relationship between nonalcoholic fatty liver disease (NAFLD), serum adiponectin concentration and brachial-ankle pulse wave velocity (baPWV) as a risk marker for atherosclerosis. METHODS: A total of 213 nonalcoholic subjects (67 males, 146 females) participated in this study. Division of subjects into the NAFLD group or the normal group was based on the existence of fatty liver detected by sonography. RESULTS: Serum adiponectin levels in the NAFLD group were significantly lower than those in the normal group. After adjusting for age, body-mass index (BMI) and the homeostasis model of assessment (HOMA), there was a significant negative correlation between NAFLD and serum adiponectin level only in females (r = -0.22, P = 0.008). Multiple logistic regression analysis showed a tendency of inverse correlation between NAFLD and serum adiponectin level in females (P = 0.055). After adjustment for age, BMI and HOMA value, serum adiponectin levels were inversely correlated with serum alanine aminotransferase (ALT) and gamma-glutamyltranspeptidase (GGT) levels (r = -0.199 (P = 0.004) and r = -0.282 (P < 0.001)). On the other hand, baPWV in the NAFLD group was also significantly higher than that in the normal group in females (P = 0.005). Individual levels of serum ALT, aspatate aminotransferase (AST), alkaline phosphatase (ALP) and GGT were positively correlated with baPWV after adjusting for age, sex, BMI, HOMA and systolic blood pressure (P < 0.05). CONCLUSION: Serum adiponectin level and baPWV were significantly associated with NAFLD and various liver enzymes, especially in females. PMID- 15745931 TI - The association of plasma adiponectin levels with hypertensive retinopathy. AB - OBJECTIVE: Previous studies have demonstrated that low plasma adiponectin concentrations are associated with essential hypertension. It has also recently been shown that adiponectin plays an essential role in the modulation of angiogenesis. These data led us to hypothesize that adiponectin might contribute to end-organ damage in hypertension. METHODS: In the present study we have evaluated the relationship between plasma adiponectin concentrations and hypertensive retinopathy. One hundred and ten patients newly diagnosed with essential hypertension (EHT) (mean age, 46.79+/-5.0 years; body mass index (BMI), 26.47+/-2.23 kg/m(2); male/female ratio, 58/52) and 57 healthy normotensive control subjects (NT) (mean age, 46.84+/-5.4 years; BMI, 26.66+/-2.65 kg/m(2); male/female ratio, 33/24) were enrolled. RESULTS: Plasma adiponectin levels were significantly lower in EHT than in NT (P < 0.001). In addition, adiponectin concentrations were strongly correlated with systolic and diastolic blood pressures in EHT (r = -0.757, P < 0.001; r = -0.761, P < 0.001) while there was no correlation in the NT group. Plasma adiponectin in patients with grade 0 hypertensive retinopathy (n = 52) was significantly higher than that of the patients with grade 1 (n = 30) and 2 (n = 28) hypertensive retinopathy (P < 0.001 for each). Plasma adiponectin in patients with grade 0 hypertensive retinopathy was also significantly lower than that in the NT group (P < 0.001). The estimated threshold of plasma adiponectin concentration for hypertensive retinopathy was 17 microg/ml. This critical adiponectin level served largely to separate patients with retinopathy from those without. CONCLUSION: Our results have shown that plasma adiponectin concentrations decrease progressively with higher grades of hypertensive retinopathy even after correction for other atherogenic risk factors, suggesting that a critical adiponectin level is needed for the development of retinopathy. PMID- 15745932 TI - Insulin-secretion abnormalities and clinical deterioration related to impaired glucose tolerance in cystic fibrosis. AB - OBJECTIVE: To evaluate insulin-secretion kinetics and insulin sensitivity in cystic fibrosis (CF) patients with normal glucose tolerance (CF-NGT), impaired glucose tolerance (CF-IGT) or CF-related diabetes (CFRD), and the potential effects of moderate hyperglycemia on clinical and nutritional status. DESIGN AND METHODS: Cross-sectional study including 50 outpatients with CF. Patients underwent both oral (OGGT) and intravenous (IVGTT) glucose tolerance tests in order to assess insulin secretion and peripheral insulin sensitivity. Homeostasis assessment model and OGGT were used to investigate insulin sensitivity. Forced expiratory volume in the first second (FEV(1)) and forced vital capacity (FVC) were measured to evaluate pulmonary function. Body mass index (BMI) was determined to assess nutritional status. RESULTS: Insulin secretion was significantly decreased (and delayed at OGTT) in the CFRD group (n = 9) versus the CF-IGT group (n = 10) and the CF-IGT versus the CF-NGT group (n = 31). Insulin sensitivity was significantly different in the CF-IGT and CFRD groups versus the CF-NGT group. FEV(1), FVC and BMI presented a significant linear correlation with plasma glucose value at 120 min at OGTT and were significantly lower in both CF-IGT and CFRD versus the CF-NGT group, whereas no differences were found between the CF-IGT and CFRD groups. CONCLUSIONS: CF patients with IGT present diminished insulin secretion and increased peripheral insulin resistance, correlating with a worse clinical status, undernutrition and impaired pulmonary function. These findings open the question of whether early treatment of mild alterations of glucose metabolism with insulin secretagogues or short-action insulin may lead to improvement of clinical status in CF patients. PMID- 15745933 TI - Is there a gender difference in the associations of birthweight and adult hypothalamic-pituitary-adrenal axis activity? AB - OBJECTIVE: Increased hypothalamic-pituitary-adrenal (HPA) axis activity in men of low birthweight may be an important link between early life and the adult metabolic syndrome. In animal models females are more sensitive than males to HPA axis programming, but whether gender influences susceptibility in humans is unknown. DESIGN: Birth cohort study. METHODS: We studied 106 women aged 67-78 years, from Hertfordshire, UK, in whom birthweight was recorded. Negative feedback sensitivity was assessed by an overnight low-dose (0.25 mg) dexa methasone suppression test, and adrenal sensitivity by a low-dose (1 microg) ACTH(1 - 24) stimulation test. Cortisol and its metabolites were analysed in a 24 h urine collection. Data were compared with previously published identical measurements in 205 men aged 66-77 years from the same cohort. RESULTS: In women, plasma cortisol levels after dexamethasone were lower (P < 0.0001) and peak cortisol following ACTH(1 - 24) were higher (P < 0.0001) than in men, suggesting a more responsive HPA axis. As in men, women with lower birthweight had enhanced plasma cortisol responses to ACTH(1 - 24) (P = 0.05 for trend) but no difference in plasma cortisol after dexamethasone or in urinary cortisol metabolite excretion. The strength of the association in women was not different from that in men; a 1 lb decrease in birthweight was associated with an incremental rise in cortisol of 12.6 nmol/l (95% confidence interval (CI) 1.4, 23.8) in men, P = 0.03, and 14.8 nmol/l (95% CI -0.4, 29.9) in women, P = 0.05 (P = 0.82 for birthweight x gender interaction). In a combined analysis of men and women adjusted for gender (n = 302), a 1 lb decrease in birthweight was associated with a 13.4 nmol/l (95% CI 4.5, 22.4) greater incremental rise in plasma cortisol, P = 0.003. CONCLUSIONS: Associations between lower birthweight and increased HPA axis activity are similar in men and women, supporting the hypothesis that HPA axis activation is an important mechanism underlying programming of adult disease. PMID- 15745934 TI - Novel insertion frameshift mutation of the LH receptor gene: problematic clinical distinction of Leydig cell hypoplasia from enzyme defects primarily affecting testosterone biosynthesis. AB - Leydig cell hypoplasia (LCH) is a rare autosomal recessive condition that interferes with normal development of male external genitalia in 46,XY individuals and is caused by inactivating mutations of the LH receptor gene. The clinical and biochemical diagnostic parameters of LCH are not always specific and may therefore show significant overlap with other causes of insufficient testicular steroid biosynthesis. We have studied a 46,XY newborn with completely female external genitalia and palpable testes. Due to an increased basal serum ratio of androstenedione/testosterone, 17 beta-hydroxysteroid dehydrogenase type 3 (17 beta-HSD 3) deficiency was initially suspected. DNA analysis of the corresponding HSD17B3 gene, however, showed no abnormalities in the entire coding region. In contrast, direct sequencing of the LH receptor gene revealed a novel homozygous single nucleotide insertion in exon 11 (codon A589fs) producing a frame shift in the open reading frame predicting for premature termination of translation 17 amino acids downstream. From the genetic perspective, this mutation represents the first frame shift mutation in the LH receptor gene ever reported to date. From the clinical standpoint, LCH should always be considered in the differential diagnosis as steroid profiles may not be informative. Therefore, molecular genetic analysis should be warranted for androgen biosynthesis defects in all cases. PMID- 15745935 TI - A direct approach to the estimation of the origin of oestrogens and androgens in elderly men by comparison with hormone levels in postmenopausal women. AB - OBJECTIVE: The origin of oestrogens in men is only partly understood. From infusion studies with radioactively labelled hormones, we know that oestradiol (E2) and oestrone (E1) are either directly secreted by the testes and adrenal glands or peripherally produced from testicular or adrenal androgens. DESIGN AND METHODS: We determined E2, E1, androstenedione, testosterone and dehydroepiandroster-one sulphate (DHEAS) in 292 elderly men and 367 postmenopausal women. We considered post-menopausal women as men without testes, assuming that the postmenopausal ovary is not endocrinologically active and that the testes do not contribute to circulating levels of DHEAS. Subjects were stratified by DHEAS levels to adjust for differences in DHEAS levels between sexes. For men and women separately, mean levels of E2, E1, androstenedione and testosterone were calculated per DHEAS stratum. The relative direct and indirect contributions of the testes to steroid levels in men were calculated by the formula [(C(m) - C(f))/C(m)] x 100%, in which C(m) and C(f) represent the mean concentrations of the steroid in men and women respectively. RESULTS: The relative contributions (%) of the testes to hormone levels per DHEAS stratum (<2, 2-4, 4-6 and >6 micromol/l) respectively were, for E2, 72%, 60%, 52% and 44%; for E1, 54%, 47%, 35% and 34%; for androstenedione, 14%, 4%, 12% and 0%; and, for testosterone, 88%, 88%, 87% and 83%. CONCLUSIONS: We conclude that in elderly men dependent on DHEAS levels, 44-72% of E2 and 34-54% of E1 originate directly or indirectly from the testes. PMID- 15745936 TI - Effects of metformin and ethinyl estradiol-cyproterone acetate on lipid levels in obese and non-obese women with polycystic ovary syndrome. AB - OBJECTIVE: Women with polycystic ovary syndrome (PCOS) exhibit risk factors for cardiovascular diseases such as abdominal obesity, insulin resistance and dyslipidemia. Insulin sensitizers, especially metformin, have been shown to improve these metabolic disturbances, but there are only a few studies on their effects on serum lipids in polycystic ovary syndrome. METHODS: Thirty-five women with PCOS (18 obese and 17 non-obese) were randomized to 6-month treatments with metformin or ethinyl estradiol-cyproterone acetate oral contraceptive pills. RESULTS: In the whole-study population (non-obese and obese women) serum levels of high-density lipoprotein cholesterol increased from 1.4+/-0.2 to 1.6+/-0.1 mmol/l (means +/-S.E. throughout) at 3 and 6 months (P < 0.001), the total cholesterol:high-density lipoprotein cholesterol ratio decreased significantly from 3.8+/-0.3 to 3.3+/-0.2 at 6 months (P < 0.001) and a similar trend was observed in serum triglyceride levels during metformin treatment. In the oral contraceptive group, serum levels of total cholesterol increased from 4.9+/-0.3 to 5.4+/-0.3 mmol/l (P < 0.05), high-density lipoprotein cholesterol increased from 1.2+/-0.1 to 1.5+/-0.1 mmol/l (P < 0.001), the total cholesterol:high density lipoprotein cholesterol ratio decreased from 4.6+/-0.4 to 3.7+/-0.2 (P < 0.001) and triglycerides increased from 1.3+/-0.1 to 1.9+/-0.2 mmol/l at 6 months of treatment (P < 0.001). Serum low-density lipoprotein cholesterol levels remained unchanged during both treatments. Milder but similar changes in the subgroups of obese and non-obese women were observed during both treatments. Moreover, in the whole-study population both systolic (P = 0.02) and diastolic (P = 0.05) blood pressures decreased over the 6 months of metformin treatment. CONCLUSION: In women with PCOS, metformin treatment had beneficial effects on lipid profile and blood pressure, and therefore it could be useful in the prevention of cardiovascular complications in these women. PMID- 15745937 TI - Inhibin alpha-subunit and the inhibin coreceptor betaglycan are downregulated in endometrial carcinoma. AB - OBJECTIVE: In the present study we evaluated the protein distribution and mRNA levels of inhibin alpha-subunit and its coreceptor betaglycan in endometrial adenocarcinoma. DESIGN: Two groups of postmenopausal women were studied: the first group had recently diagnosed endometrial adenocarcinoma (n = 16; age range 61-79 years), and the second group (n = 12; age range 64-78 years) had undergone hysterectomy for uterine prolapse and served as control. METHODS: Inhibin alpha subunit and betaglycan gene expression and tissue distribution were evaluated by semiquantitative RT-PCR and immunohistochemistry respectively. RESULTS: Inhibin alpha-subunit and betaglycan mRNAs were expressed by both healthy and tumoral endometria, but their expression was significantly lower in endometrial carcinoma (P < 0.001, based on Student's t test). Inhibin alpha-subunit expression was much weaker in the glands of tumours than in non-neoplastic specimens. Betaglycan protein was identified in the epithelial cells lining non-tumoral endometrium, and in endothelial cells of both normal and tumoral endometria. Well differentiated neoplastic cells had a faint and scarce betaglycan staining, and poorly differentiated cells did not express betaglycan at all. CONCLUSIONS: The lower inhibin alpha and betaglycan expression in endometrial adenocarcinoma suggests that the inhibin action may be disrupted. However, the expression of betaglycan in the endothelia of the tumour vasculature suggests that a selective vascular response to inhibin may be possible in these tumours. PMID- 15745938 TI - Testosterone does not adversely affect fibrinogen or tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) levels in 46 men with chronic stable angina. AB - OBJECTIVE: In women, sex hormones cause increased morbidity and mortality in patients with coronary heart disease (CHD) and adversely affect the coagulation profile. We have studied the effect of physiological testosterone replacement therapy in men on coagulation factor expression, to determine if there is an increased risk of thrombosis. METHODS: Double-blind, randomized, placebo controlled trial of testosterone in 46 men with chronic stable angina. Measurements of free, total and bioavailable testosterone, luteinising hormone (LH) and follicle-stimulating hormone (FSH), estradiol, plasminogen activator inhibitor-1 (PAI-1), fibrinogen, tissue plasminogen activator (tPA) and full blood count were made at 0, 6 and 14 weeks. RESULTS: Bioavailable testosterone levels were: 2.58 +/- 0.58 nmol/l at baseline, compared with 3.35 +/- 0.31 nmol/l at week 14 (P < 0.001) after treatment compared with 2.6 +/- 0.18 nmol/l and 2.44 +/- 0.18 nmol/l in the placebo group (P was not significant). There was no change in fibrinogen (3.03 +/- 0.18 g/l at baseline and 3.02 +/- 0.18 g/l at week 14, P = 0.24), tPA activity (26.77 +/- 4.9 Iu/ml and 25.67 +/- 4.4 Iu/ml, P = 0.88) or PAI-1 activity (0.49 +/- 0.85 Iu/ml and 0.36 +/- 0.06 Iu/ml, P = 0.16) with active treatment and no differences between the groups (at week 14, P value 0.98, 0.59 and 0.8 for fibrinogen, PAI-1 and tPA respectively). Haemoglobin concentration did not change over time, in the testosterone group (1.44 +/- 0.02 g/l and 1.45 +/- 0.02 g/l, P = 0.22). CONCLUSION: Physiological testosterone replacement does not adversely affect blood coagulation status. PMID- 15745939 TI - Correlation of VEGF production with IL1 alpha and IL6 secretion by human pituitary adenoma cells. AB - OBJECTIVES: Vascular endothelial growth factor (VEGF) is considered to be the most important angiogenic factor involved in the neovascularisation of solid tumours. Regulatory molecules include cytokines and growth factors. Interleukin (IL)1 and IL6 have both been shown to regulate VEGF levels in a variety of tissues. The role of cytokines in the pathogenesis of pituitary tumours remains unclear. We have examined the expression of VEGF and its relationships with IL1 and IL6 in the human pituitary tumour cell line HP75 and a series of human pituitary tumours. We have also looked at the relationship of tumour volume and invasive status to VEGF secretion. METHODS: Surgically resected tumours were routinely cultured in single-cell suspension at 200 K/well (standard unit for culture of dispersed primary pituitary adenoma cells). We measured VEGF, IL1 alpha and IL6 levels by ELISA. Tumour volume and invasion grade were assessed by preoperative magnetic resonance imaging. RESULTS: VEGF was detected in conditioned medium of HP75 cells (900+/-52 pg/ml) and in 82% of tumours tested (range 26-16 464 pg/ml). Tumour volume and secretion of VEGF were significantly associated with levels of IL6 (volume, P = 0.056; VEGF, P < 0.001 (P values based on Spearman's test)) and IL1 alpha produced (volume, P < 0.005; VEGF, P < 0.001). Invasive tumours showed a higher basal secretion of VEGF that that of the non invasive type; however, this difference was not significant. Addition of exogenous IL1 alpha, but not IL6, significantly increased VEGF production. CONCLUSIONS: The significant associations between VEGF and the levels of IL6 and IL1 alpha suggest an important role for these cytokines in the development of these tumours. PMID- 15745940 TI - Estrogen responsiveness of bone formation in vitro and altered bone phenotype in aged estrogen receptor-alpha-deficient male and female mice. AB - OBJECTIVE: Although the beneficial effects of estrogen on bone are well known, the roles of estrogen receptors (ERs) in mediating these effects are not fully understood. METHODS: To study the effects of long-term ER alpha deficiency, bone phenotype was studied in aged ER alpha knockout (ERKO) mice. In addition, ERKO osteoclasts and osteoblasts were cultured in vitro. DESIGN AND RESULTS: Histomorphometric analysis showed that the trabecular bone volume and thickness were significantly increased and the rate of bone formation enhanced in both male and female ERKO mice in comparison to the wild-type animals. In ERKO males, however, the bones were thinner and their maximal bending strengths decreased. Consistent with previous reports, the bones of knockout mice, especially of female mice, were shorter than those of wild-type mice. In addition, the growth plates were totally absent in the tibiae of aged ERKO females, whereas the growth plate cartilages were detectable in wild-type females as well as in all the males. Analysis of cultured bone marrow cells from 10- to 12-week-old mice demonstrated that 17 beta-estradiol could stimulate osteoblastic differentiation of bone marrow cells derived from ERKO mice relatively to the same extent as those derived from wild-type mice. This was demonstrated by increases in synthesis of type I collagen, activity of alkaline phosphatase and accumulation of calcium in cultures. Total protein content was, however, reduced in ERKO osteoblast cultures. CONCLUSIONS: These results show altered bone phenotype in ERKO mice and demonstrate the stimulatory effect of estrogen on osteoblasts even in the absence of full-length ER alpha. PMID- 15745941 TI - The selective prostaglandin EP4 agonist, APS-999 Na, induces follicular growth and maturation in the rat ovary. AB - OBJECTIVE: It is known that prostaglandin E2 (PGE2) plays a crucial role in the ovulation process. PGE2 mediates its actions through four subtypes of receptors designated EP1-EP4. However, the nature of the ovulation-promoting effect of PGE2 is not well understood. We have carried out this in vivo research utilizing a new selective EP4 receptor agonist (EP4A; APS-999 Na) to investigate its role in folliculogenesis aiming at more understanding of the biological mechanism of action of PGE2. DESIGN AND METHODS: Immature, 22 day old Wistar rats were used. Animals were injected once either with placebo, 20 IU pregnant mare serum gonadotrophin (PMSG), or EP4A (10, 20 or 50 microg). In other experiments, EP4A was injected in PMSG stimulated rats. Ovarian follicle growth and development was assessed through total count and size measurement of ovulatory follicles in whole ovaries of the investigated groups. Utilizing immunohistochemistry, the spatial localization of the EP4 receptor in immature rat ovary and IL-8 pattern of expression after EP4A injection was also investigated. RESULTS: Our study showed that injection of EP4A resulted in increased follicle growth and development compared with the control, in a time- and dose-dependent manner. The highest values for follicle count and size were observed 12 and 24 h after EP4A injection. EP4A induced follicle growth with morphological characteristics similar to that induced by the standard PMSG. Most dense immune staining for EP4 receptor was noticed in proliferating granulosa cells of the secondary follicles while those of the primordial and primary follicles were not stained. EP4A injection induced the expression of IL-8 in the follicles. CONCLUSIONS: Our study revealed that: (i) the localization of EP4 receptor in the ovary implies a role in follicle growth, (ii) PGE2 induced ovarian follicle growth and development is mediated at least in part through the EP4 receptor, (iii) the action of EP4A is mediated through IL-8 up regulation and (iv) the new EP4A could be a promising reagent for various systems used to induce follicle maturation in clinical or agricultural fields. This knowledge may provide useful targets for manipulation of infertility. PMID- 15745942 TI - Afferents and homotypic neighbors regulate horizontal cell morphology, connectivity, and retinal coverage. AB - Horizontal cells are inhibitory interneurons with laterally oriented dendrites that overlap one another, contacting the pedicles of cone photoreceptors. Because of their regular spacing, the network of horizontal cells provides a uniform coverage of the retinal surface. The developmental processes establishing these network properties are undefined, but cell-intrinsic instructions and interactions with other cells have each been suggested to play a role. Here, we show that the intercellular spacing of horizontal cells is essentially independent of genetic background and is predicted by local density, suggesting that horizontal cell positioning is modulated by proximity to other horizontal cells. Dendritic field area compensates for this variation in intercellular spacing, maintaining constant dendritic coverage between strains. Functional dendritic overlap is achieved anatomically at the level of the pedicles, where horizontal cells interact with one another to establish their connectivity: the number of dendritic terminals contacting a pedicle changes, reciprocally, between neighboring horizontal cells during development based on their relative proximity to each pedicle. Cellular morphology is also shown to be regulated by the afferents themselves: afferent elimination before innervation does not alter dendritic field size nor stratification but compromises dendritic branching and prevents terminal formation. Afferent and homotypic interactions therefore generate the morphology, spacing, and connectivity of horizontal cells underlying their functional coverage of the retina. PMID- 15745943 TI - Constitutional aneuploidy in the normal human brain. AB - The mouse brain contains genetically distinct cells that differ with respect to chromosome number manifested as aneuploidy (Rehen et al., 2001); however, the relevance to humans is not known. Here, using double-label fluorescence in situ hybridization for the autosome chromosome 21 (chromosome 21 point probes combined with chromosome 21 "paint" probes), along with immunocytochemistry and cell sorting, we present evidence for chromosome gain and loss in the human brain. Chromosome 21 aneuploid cells constitute approximately 4% of the estimated one trillion cells in the human brain and include non-neuronal cells and postmitotic neurons identified by the neuronspecific nuclear protein marker. In comparison, human interphase lymphocytes present chromosome 21 aneuploidy rates of 0.6%. Together, these data demonstrate that human brain cells (both neurons and non neuronal cells) can be aneuploid and that the resulting genetic mosaicism is a normal feature of the human CNS. PMID- 15745944 TI - Trafficking of a ligand-receptor complex on the growth cones as an essential step for the uptake of nerve growth factor at the distal end of the axon: a single molecule analysis. AB - The behavior of single molecules of neurotrophins on growth cones was observed by the use of the fluorescent conjugate of nerve growth factor (NGF), Cy3-NGF. After the application of 0.4 nm Cy3-NGF, chick dorsal root ganglion growth cones responded within 1 min of adding the stimulus by expanding their lamellipodia. Only 40 molecules of Cy3-NGF, which occupied <5% of the estimated total binding sites on a single growth cone, were required to initiate the motile responses. After binding to the high-affinity receptor, Cy3-NGF displayed lateral diffusion on the membrane of the growth cones with a diffusion constant of 0.3 microm2 s( 1). The behavior of Cy3-NGF was shifted to a one-directional rearward movement toward the central region of the growth cone. The one-directional movement of Cy3 NGF displayed the same rate as the rearward flow of actin, approximately 4 microm/min. This movement could be stopped by the application of the potent inhibitor of actin polymerization, latrunculin B. Molecules of Cy3-NGF were suggested to be internalized in the vicinity of the central region of the growth cone during this rearward trafficking, because Cy3-NGF remained in the growth cone after the growth cones had been exposed to an acidic surrounding medium: acidic medium causes the complete dissociation of Cy3-NGF from the receptors on the surface of growth cones. These results suggested that actin-driven trafficking of the NGF receptor complex is an essential step for the accumulation and endocytosis of NGF at the growth cone and for the retrograde transport of NGF toward the cell body. PMID- 15745945 TI - Properties of synaptically evoked astrocyte calcium signal reveal synaptic information processing by astrocytes. AB - The synaptic control of the astrocytic intracellular Ca2+ is crucial in the reciprocal astrocyte-neuron communication. Using electrophysiological and Ca2+ imaging techniques in rat hippocampal slices, we investigated the astrocytic Ca2+ signal modulation induced by synaptic terminals that use glutamate and acetylcholine. Ca2+ elevations were evoked by glutamate released from Schaffer collaterals and by acetylcholine, but not glutamate, released by alveus stimulation, indicating that astrocytes discriminate the activity of different synapses belonging to different axon pathways. The Ca2+ signal was modulated bidirectionally by simultaneous activation of both pathways, being depressed at high stimulation frequencies and enhanced at low frequencies. The Ca2+ modulation was attributable to astrocytic intrinsic properties, occurred at discrete regions of the processes, and controlled the intracellular expansion of the Ca2+ signal. In turn, astrocyte Ca2+ signal elicited NMDA receptor-mediated currents in pyramidal neurons. Therefore, because astrocytes discriminate and integrate synaptic information, we propose that they can be considered as cellular elements involved in the information processing by the nervous system. PMID- 15745946 TI - The multiple functions of cysteine-string protein analyzed at Drosophila nerve terminals. AB - The synaptic vesicle-associated cysteine-string protein (CSP) is important for synaptic transmission. Previous studies revealed multiple defects at neuromuscular junctions (NMJs) of csp null-mutant Drosophila, but whether these defects are independent of each other or mechanistically linked through J domain mediated-interactions with heat-shock cognate protein 70 (Hsc70) has not been established. To resolve this issue, we genetically dissected the individual functions of CSP by an in vivo structure/function analysis. Expression of mutant CSP lacking the J domain at csp null-mutant NMJs fully restored normal thermo tolerance of evoked transmitter release but did not completely restore evoked release at room temperature and failed to reverse the abnormal intraterminal Ca2+ levels. This suggests that J domain-mediated functions are essential for the regulation of intraterminal Ca2+ levels but only partially required for regulating evoked release and not required for protecting evoked release against thermal stress. Hence, CSP can also act as an Hsc70-independent chaperone protecting evoked release from thermal stress. Expression of mutant CSP lacking the L domain restored neurotransmission and partially reversed the abnormal intraterminal Ca2+ levels, suggesting that the L domain is important, although not essential, for the role of CSP in regulating intraterminal Ca2+ levels. We detected no effects of csp mutations on individual presynaptic Ca2+ signals triggered by action potentials, suggesting that presynaptic Ca2+ entry is not primarily impaired. Both the J and L domains were also required for the role of CSP in synaptic growth. Together, these results suggest that CSP has several independent synaptic functions, affecting synaptic growth, evoked release, thermal protection of evoked release, and intraterminal Ca2+ levels at rest and during stimulation. PMID- 15745947 TI - Glutamate receptor trafficking: endoplasmic reticulum quality control involves ligand binding and receptor function. AB - The glutamate receptor (GluR) agonist-binding site consists of amino acid residues in the extracellular S1 and S2 domains in the N-terminal and M3-M4 loop regions, respectively. In the present study, we sought to confirm that the conserved ligand-binding residues identified in the AMPA receptor S1S2 domains also participate in ligand binding of GluR6 kainate receptors. Amino acid substitutions were made in the GluR6 parent at R523, T690, and E738 to alter their potential interactions with ligand. Mutant receptors were expressed in human embryonic kidney 293 cells, confirmed by Western blot analysis, and tested by [3H]kainate binding and patch-clamp recording. Each of the binding site mutations was sufficient to reduce [3H]kainate binding to undetectable levels and eliminate functional responses to glutamate or kainate. As with our studies of other nonfunctional mutants (Fleck et al., 2003), immunocytochemical staining and cell-surface biotinylation studies showed that the mutant receptors were retained intracellularly and did not traffic to the cell surface. Endoglycosidase-H digests and colocalization with endoplasmic reticulum (ER) markers demonstrated that the mutant receptors are immaturely glycosylated and retained in the ER. Immunoprecipitation, native PAGE, and functional studies confirmed that the GluR6 binding site mutants are capable of multimeric assembly, indicating their retention in the ER does not result from a gross protein folding error. Together, these results confirm the role of R523, T690, and E738 directly in ligand binding to GluR6 and further support our previous report that nonfunctional GluRs are retained intracellularly by a functional checkpoint in ER quality control. PMID- 15745948 TI - Regulation of motoneuron excitability via motor endplate acetylcholine receptor activation. AB - Motoneuron populations possess a range of intrinsic excitability that plays an important role in establishing how motor units are recruited. The fact that this range collapses after axotomy and does not recover completely until after reinnervation occurs suggests that muscle innervation is needed to maintain or regulate adult motoneuron excitability, but the nature and identity of underlying mechanisms remain poorly understood. Here, we report the results of experiments in which we studied the effects on rat motoneuron excitability produced by manipulations of neuromuscular transmission and compared these with the effects of peripheral nerve axotomy. Inhibition of acetylcholine release from motor terminals for 5-6 d with botulinum toxin produced relatively minor changes in motoneuron excitability compared with the effect of axotomy. In contrast, the blockade of acetylcholine receptors with alpha-bungarotoxin over the same time interval produced changes in motoneuron excitability that were statistically equivalent to axotomy. Muscle fiber recordings showed that low levels of acetylcholine release persisted at motor terminals after botulinum toxin, but endplate currents were completely blocked for at least several hours after daily intramuscular injections of alpha-bungarotoxin. We conclude that the complete but transient blockade of endplate currents underlies the robust axotomy-like effects of alpha-bungarotoxin on motoneuron excitability, and the low level of acetylcholine release that remains after injections of botulinum toxin inhibits axotomy-like changes in motoneurons. The results suggest the existence of a retrograde signaling mechanism located at the motor endplate that enables expression of adult motoneuron excitability and depends on acetylcholine receptor activation for its normal operation. PMID- 15745949 TI - Compartment-resolved imaging of activity-dependent dynamics of cortical blood volume and oximetry. AB - Optical imaging, positron emission tomography, and functional magnetic resonance imaging (fMRI) all rely on vascular responses to image neuronal activity. Although these imaging techniques are used successfully for functional brain mapping, the detailed spatiotemporal dynamics of hemodynamic events in the various microvascular compartments have remained unknown. Here we used high resolution optical imaging in area 18 of anesthetized cats to selectively explore sensory-evoked cerebral blood-volume (CBV) changes in the various cortical microvascular compartments. To avoid the confounding effects of hematocrit and oximetry changes, we developed and used a new fluorescent blood plasma tracer and combined these measurements with optical imaging of intrinsic signals at a near isosbestic wavelength for hemoglobin (565 nm). The vascular response began at the arteriolar level, rapidly spreading toward capillaries and venules. Larger veins lagged behind. Capillaries exhibited clear blood-volume changes. Arterioles and arteries had the largest response, whereas the venous response was smallest. Information about compartment-specific oxygen tension dynamics was obtained in imaging sessions using 605 nm illumination, a wavelength known to reflect primarily oximetric changes, thus being more directly related to electrical activity than CBV changes. Those images were radically different: the response began at the parenchyma level, followed only later by the other microvascular compartments. These results have implications for the modeling of fMRI responses (e.g., the balloon model). Furthermore, functional maps obtained by imaging the capillary CBV response were similar but not identical to those obtained using the early oximetric signal, suggesting the presence of different regulatory mechanisms underlying these two hemodynamic processes. PMID- 15745951 TI - Ethanol-responsive brain region expression networks: implications for behavioral responses to acute ethanol in DBA/2J versus C57BL/6J mice. AB - Activation of the mesolimbic dopamine reward pathway by acute ethanol produces reinforcement and changes in gene expression that appear to be crucial to the molecular basis for adaptive behaviors and addiction. The inbred mouse strains DBA/2J and C57BL/6J exhibit contrasting acute behavioral responses to ethanol. We used oligonucleotide microarrays and bioinformatics methods to characterize patterns of gene expression in three brain regions of the mesolimbic reward pathway of these strains. Expression profiling included examination of both differences in gene expression 4 h after saline injection or acute ethanol (2 g/kg). Using a rigorous stepwise method for microarray analysis, we identified 788 genes differentially expressed in control DBA/2J versus C57BL/6J mice and 307 ethanol-regulated genes in the nucleus accumbens, prefrontal cortex, and ventral tegmental area. There were strikingly divergent patterns of ethanol-responsive gene expression in the two strains. Ethanol-responsive genes also showed clustering at discrete chromosomal regions, suggesting local chromatin effects in regulation. Ethanol-regulated genes were generally related to neuroplasticity, but regulation of discrete functional groups and pathways was brain region specific: glucocorticoid signaling, neurogenesis, and myelination in the prefrontal cortex; neuropeptide signaling and developmental genes, including factor Bdnf, in the nucleus accumbens; and retinoic acid signaling in the ventral tegmental area. Bioinformatics analysis identified several potential candidate genes for quantitative trait loci linked to ethanol behaviors, further supporting a role for expression profiling in identifying genes for complex traits. Brain region-specific changes in signaling and neuronal plasticity may be critical components in development of lasting ethanol behavioral phenotypes such as dependence, sensitization, and craving. PMID- 15745950 TI - Interactions between ephrin-B and metabotropic glutamate 1 receptors in brain tissue and cultured neurons. AB - We examined the interaction between ephrins and metabotropic glutamate (mGlu) receptors in the developing brain and cultured neurons. EphrinB2 coimmunoprecipitated with mGlu1a receptors, in all of the brain regions examined, and with mGlu5 receptors in the corpus striatum. In striatal slices, activation of ephrinB2 by a clustered form of its target receptor, EphB1, amplified the mGlu receptor-mediated stimulation of polyphosphoinositide (PI) hydrolysis. This effect was abolished in slices treated with mGlu1 or NMDA receptor antagonists but was not affected by pharmacological blockade of mGlu5 receptors. An interaction among ephrinB2, mGlu1 receptor, and NMDA was supported by the following observations: (1) the NR1 subunit of NMDA receptors coimmunoprecipitated with mGlu1a receptors and ephrinB2 in striatal lysates; (2) clustered EphB1 amplified excitatory amino acid-stimulated PI hydrolysis in cultured granule cells grown under conditions that favored the expression of mGlu1a receptors; and (3) clustered EphB1 amplified the enhancing effect of mGlu receptor agonists on NMDA toxicity in cortical cultures, and its action was sensitive to mGlu1 receptor antagonists. Finally, fluorescence resonance energy transfer and coclustering analysis in human embryonic kidney 293 cells excluded a physical interaction between ephrinB2 and mGlu1a (or mGlu5 receptors). A functional interaction between ephrinB and mGlu1 receptors, which likely involves adaptor or scaffolding proteins, might have an important role in the regulation of developmental plasticity. PMID- 15745952 TI - Insulin-like growth factor-1 inhibits adult supraoptic neurons via complementary modulation of mechanoreceptors and glycine receptors. AB - In the CNS, insulin-like growth factor-1 (IGF-1) is mainly known for its trophic effect both during development and in adulthood. Here, we show than in adult rat supraoptic nucleus (SON), IGF-1 receptor immunoreactivity is present in neurons, whereas IGF-1 immunoreactivity is found principally in astrocytes and more moderately in neurons. In vivo application of IGF-1 within the SON acutely inhibits the activity of both vasopressin and oxytocin neurons, the two populations of SON neuroendocrine cells. Recordings of acutely isolated SON neurons showed that this inhibition occurs through two rapid and reversible mechanisms, both involving the neuronal IGF-1 receptor but different intracellular messengers. IGF-1 inhibits Gd3+-sensitive and osmosensitive mechanoreceptor cation current via phosphatidylinositol-3 (PI3) kinase activation. IGF-1 also potentiates taurine-activated glycine receptor (GlyR) Cl- currents by increasing the agonist sensitivity through a extremely rapid (within a second) PI3 kinase-independent mechanism. Both mechanoreceptor channels and GlyR, which form the excitatory and inhibitory components of SON neuron osmosensitivity, are active at rest, and their respective inhibition and potentiation will both be inhibitory, leading to strong decrease in neuronal activity. It will be of interest to determine whether IGF-1 is released by neurons, thus participating in an inhibitory autocontrol, or astrocytes, then joining the growing family of glia-to-neuron transmitters that modulate neuronal and synaptic activity. Through the opposite and complementary acute regulation of mechanoreceptors and GlyR, IGF-1 appears as a new important neuromodulator in the adult CNS, participating in the complex integration of neural messages that regulates the level of neuronal excitability. PMID- 15745953 TI - Role of the primary motor and dorsal premotor cortices in the anticipation of forces during object lifting. AB - When lifting small objects, people apply forces that match the expected weight of the object. This expectation relies in part on information acquired during a previous lift and on associating a certain weight with a particular object. Our study examined the role of the primary motor and dorsal premotor cortices in predicting weight based either on information acquired during a previous lift (no cue experiment) or on arbitrary color cues associated with a particular weight (cue experiment). In the two experiments, subjects used precision grip to lift two different weights in a series of trials both before and after we applied low frequency repetitive transcranial magnetic stimulation over the primary motor and dorsal premotor cortices. In the no-cue experiment, subjects did not receive any previous information about which of two weights they would have to lift. In the cue experiment, a color cue provided information about which of the two weights subjects would have to lift. Our results demonstrate a double dissociation in the effects induced by repetitive stimulation. When applied over the primary motor cortex, repetitive stimulation disrupted the scaling of forces based on information acquired during a previous lift. In contrast, when applied over the dorsal premotor cortex, repetitive stimulation disrupted the scaling of forces based on arbitrary color cues. We conclude that the primary motor and dorsal premotor cortices have unique roles during the anticipatory scaling of forces associated with the lifting of different weights. PMID- 15745954 TI - Neurosteroid-induced plasticity of immature synapses via retrograde modulation of presynaptic NMDA receptors. AB - Neurosteroids are produced de novo in neuronal and glial cells, which begin to express steroidogenic enzymes early in development. Studies suggest that neurosteroids may play important roles in neuronal circuit maturation via autocrine and/or paracrine actions. However, the mechanism of action of these agents is not fully understood. We report here that the excitatory neurosteroid pregnenolone sulfate induces a long-lasting strengthening of AMPA receptor mediated synaptic transmission in rat hippocampal neurons during a restricted developmental period. Using the acute hippocampal slice preparation and patch clamp electrophysiological techniques, we found that pregnenolone sulfate increases the frequency of AMPA-mediated miniature excitatory postsynaptic currents in CA1 pyramidal neurons. This effect could not be observed in slices from rats older than postnatal day 5. The mechanism of action of pregnenolone sulfate involved a short-term increase in the probability of glutamate release, and this effect is likely mediated by presynaptic NMDA receptors containing the NR2D subunit, which is transiently expressed in the hippocampus. The increase in glutamate release triggered a long-term enhancement of AMPA receptor function that requires activation of postsynaptic NMDA receptors containing NR2B subunits. Importantly, synaptic strengthening could also be triggered by postsynaptic neuron depolarization, and an anti-pregnenolone sulfate antibody scavenger blocked this effect. This finding indicates that a pregnenolone sulfate-like neurosteroid is a previously unrecognized retrograde messenger that is released in an activity-dependent manner during development. PMID- 15745955 TI - ets-2 promotes the activation of a mitochondrial death pathway in Down's syndrome neurons. AB - Down's syndrome (DS) is characterized by mental retardation and development of Alzheimer's disease (AD). Oxidative stress and mitochondrial dysfunction are both related to neurodegeneration in DS. Several genes in chromosome 21 have been linked to neuronal death, including the transcription factor ets-2. Cortical cultures derived from normal and DS fetal brains were used to study the role of ets-2 in DS neuronal degeneration. ets-2 was expressed in normal human cortical neurons (HCNs) and was markedly upregulated by oxidative stress. When overexpressed in normal HCNs, ets-2 induced a stereotyped sequence of apoptotic changes leading to neuronal death. DS HCNs exhibit intracellular oxidative stress and increased apoptosis after the first week in culture (Busciglio and Yankner, 1995). ets-2 levels were increased in DS HCNs, and, between 7 and 14 d in vitro, DS HCNs showed increased bax, cytoplasmic translocation of cytochrome c and apoptosis inducing factor, and active caspases 3 and 7, consistent with activation of an apoptotic mitochondrial death pathway. Degeneration of DS neurons was reduced by dominant-negative ets-2, suggesting that increased ets-2 expression promotes DS neuronal apoptosis. In the human brain, ets-2 expression was found in neurons and astrocytes. Strong ets-2 immunoreactivity was observed in DS/AD and sporadic AD brains associated with degenerative markers such as bax, intracellular Abeta, and hyperphosphorylated tau. Thus, in DS/AD and sporadic AD brains, converging pathological mechanisms leading to chronic oxidative stress and ets-2 upregulation in susceptible neurons may result in increased vulnerability by promoting the activation of a mitochondrial-dependent proapoptotic pathway of cell death. PMID- 15745957 TI - Spike-frequency adaptation separates transient communication signals from background oscillations. AB - Spike-frequency adaptation is a prominent feature of many neurons. However, little is known about its computational role in processing behaviorally relevant natural stimuli beyond filtering out slow changes in stimulus intensity. Here, we present a more complex example in which we demonstrate how spike-frequency adaptation plays a key role in separating transient signals from slower oscillatory signals. We recorded in vivo from very rapidly adapting electroreceptor afferents of the weakly electric fish Apteronotus leptorhynchus. The firing-frequency response of electroreceptors to fast communication stimuli ("small chirps") is strongly enhanced compared with the response to slower oscillations ("beats") arising from interactions of same-sex conspecifics. We are able to accurately predict the electroreceptor afferent response to chirps and beats, using a recently proposed general model for spike-frequency adaptation. The parameters of the model are determined for each neuron individually from the responses to step stimuli. We conclude that the dynamics of the rapid spike frequency adaptation is sufficient to explain the data. Analysis of additional data from step responses demonstrates that spike-frequency adaptation acts subtractively rather than divisively as expected from depressing synapses. Therefore, the adaptation dynamics is linear and creates a high-pass filter with a cutoff frequency of 23 Hz that separates fast signals from slower changes in input. A similar critical frequency is seen in behavioral data on the probability of a fish emitting chirps as a function of beat frequency. These results demonstrate how spike-frequency adaptation in general can facilitate extraction of signals of different time scales, specifically high-frequency signals embedded in slower oscillations. PMID- 15745956 TI - NMDA receptor-dependent regulation of axonal and dendritic branching. AB - In the rodent trigeminal principal nucleus (PrV), trigeminal afferent terminals and postsynaptic cells form discrete modules ("barrelettes") that replicate the patterned array of whiskers and sinus hairs on the snout. Barrelette neurons of the PrV relay whisker-specific patterns to the contralateral thalamus and, subsequently, to the primary somatosensory barrel cortex. Genetic impairment of NMDA receptor (NMDAR) function blocks development of barrelettes in the PrV. Underlying cellular and functional defects are not known. Here, we examined morphological differentiation of whisker afferents, dendritic differentiation of barrelette cells, and their electrophysiological properties in mice with genetic perturbations of the essential subunit NR1 of NMDARs. We show that in NR1 gene knock-down (KD) and knock-out mice, whisker afferents begin their embryonic development normally but, over time, fail to segregate into patches, and instead they develop exuberant terminal arbors spanning most of the PrV. Postnatal NR1KD barrelette cells, with significantly reduced NMDA currents, retain their membrane and synaptic properties but develop longer dendrites with no orientation preference. These results indicate that NMDARs regulate growth of presynaptic terminal arbors and postsynaptic dendritic branching, thereby leading to consolidation of synapses and patterning of presynaptic and postsynaptic elements. PMID- 15745958 TI - Noradrenaline and dopamine elevations in the rat prefrontal cortex in spatial working memory. AB - The role of prefrontal cortical dopamine (DA) in the modulation of working memory functions is well documented, but substantial evidence indicates that the locus ceruleus noradrenergic system also modulates working memory via actions within the prefrontal cortex (PFC). This study shows that PFC noradrenaline (NA) and DA dialysate levels phasically increase when rats perform correctly in a delayed alternation task in a T-maze, a test of spatial working memory. However, NA levels were markedly enhanced in animals trained to alternate compared with rats that acquired the spatial information about the location of food in the maze but were untrained to make a choice to obtain the reward. In contrast, PFC DA elevations occurred independently of whether the animal had acquired the trial specific information for correct task execution. The contribution of anticipatory responses to catecholamine efflux was also evaluated by exposing rats to an environment signaling the presence of the reward in the successive alternation task. No conditioned NA efflux was observed in either group. In contrast, in both groups, DA efflux increased in the anticipatory phase of the test to the same levels of those reached during the task. These data provide the first direct evidence for a selective activation of PFC NA transmission during a spatial working memory task. We propose that, in the working memory task, DA is primarily associated with reward expectancy, whereas NA is involved in the active maintenance of the information about a goal and the rules to achieve it. PMID- 15745959 TI - Neural substrates for behaviorally conditioned immunosuppression in the rat. AB - We have previously demonstrated behaviorally conditioned immunosuppression using cyclosporin A as an unconditioned stimulus and saccharin as a conditioned stimulus. In the current study, we examined the central processing of this phenomenon generating excitotoxic lesions before and after acquisition to discriminate between learning and memory processes. Three different brain areas were analyzed: insular cortex (IC), amygdala (Am), and ventromedial nucleus of the hypothalamus (VMH). The results demonstrate that IC lesions performed before and after acquisition disrupted the behavioral component of the conditioned response (taste aversion). In contrast, Am and VMH lesions did not affect conditioned taste aversion. The behaviorally conditioned suppression of splenocyte proliferation and cytokine production (interleukin-2 and interferon gamma) was differentially affected by the excitotoxic lesions, showing that the IC is essential to acquire and evoke this conditioned response of the immune system. In contrast, the Am seems to mediate the input of visceral information necessary at the acquisition time, whereas the VMH appears to participate within the output pathway to the immune system necessary to evoke the behavioral conditioned immune response. The present data reveal relevant neural mechanisms underlying the learning and memory processes of behaviorally conditioned immunosuppression. PMID- 15745961 TI - Shal and shaker differential contribution to the K+ currents in the Drosophila mushroom body neurons. AB - Shaker, a voltage-dependent K+ channel, is enriched in the mushroom bodies (MBs), the locus of olfactory learning in Drosophila. Mutations in the shaker locus are known to alter excitability, neurotransmitter release, synaptic plasticity, and olfactory learning. However, a direct link of Shaker channels to MB intrinsic neuron (MBN) physiology has not been documented. We found that transcripts for shab, shaw, shaker, and shal, among which only Shaker and Shal have been reported to code for A-type currents, are present in the MBs. The electrophysiological data showed that the absence of functional Shaker channels modifies the distribution of half-inactivation voltages (V(i1/2)) in the MBNs, indicating a segregation of Shaker channels to only a subset (approximately 28%) of their somata. In harmony with this notion, we found that approximately one-fifth of MBNs lacking functional Shaker channels displayed dramatically slowed-down outward current inactivation times and reduced peak-current amplitudes. Furthermore, whereas all MBNs were sensitive to 4-aminopyridine, a nonspecific A type current blocker, a subset of neurons (approximately 24%) displayed little sensitivity to a Shal-specific toxin. This subset of neurons displaying toxin insensitive outward currents had more depolarized V(i1/2) values attributable to Shaker channels. Our findings provide the first direct evidence that altered Shaker channel function disrupts MBN physiology in Drosophila. To our surprise, the experimental data also indicate that Shaker channels segregate to a minor fraction of MB neuronal somata (20-30%), and that Shal channels contribute the somatic A-type current in the majority of MBNs. PMID- 15745960 TI - Ca2+-dependent regulation of rho GTPases triggers turning of nerve growth cones. AB - Cytoplasmic Ca2+ elevation and changes in Rho GTPase activity are both known to mediate axon guidance by extracellular factors, but the causal relationship between these two events has been unclear. Here we show that direct elevation of cytoplasmic Ca2+ by extracellular application of a low concentration of ryanodine, which activated Ca2+ release from intracellular stores, upregulated Cdc42/Rac, but downregulated RhoA, in cultured cerebellar granule cells and human embryonic kidney 293T cells. Chemoattractive turning of the growth cone triggered by a gradient of ryanodine was blocked by overexpression of mutant forms of Cdc42 but not of RhoA in Xenopus spinal cord neurons. Furthermore, Ca2+-induced GTPase activity correlated with activation of protein kinase C and required a basal activity of Ca2+/calmodulin-dependent protein kinase II. Thus, Rho GTPases may mediate axon guidance by linking upstream Ca2+ signals triggered by guidance factors to downstream cytoskeletal rearrangements. PMID- 15745962 TI - Impaired outcome-specific devaluation of instrumental responding in mice with a targeted deletion of the AMPA receptor glutamate receptor 1 subunit. AB - The present study evaluated the proposal that mice with a targeted deletion of the glutamate receptor 1 (GluR1) subunit of the AMPA receptor are impaired in using an instrumental or pavlovian signal to gain access to a representation of the sensory-specific motivational properties of a primary reward. In experiment 1, mice were trained to approach two goal boxes in a plus-maze; each goal box contained a different reward (sucrose solution vs food pellet). After acquisition, one of the rewards was devalued by an outcome-specific satiety procedure. Subsequent test trials performed in extinction showed an increase in the latency to enter the devalued goal arm, relative to the nondevalued goal arm in control but not GluR1-/- mice. In experiment 2, a similar outcome-specific satiety procedure was used to examine the effects of reward devaluation on an instrumental nose-poke response. During testing, control but not GluR1-/- mice decreased their rate of responding on a nose poke associated with a devalued reward. A subsequent choice test showed that GluR1-/- mice were able to discriminate between the devalued and nondevalued outcomes used in both experiments. These deficits mirror those seen after lesions of the basolateral amygdala and suggests that GluR1-mediated neurotransmission in this region contributes to encoding the relationship between sensory-specific aspects of reward and their incentive value. PMID- 15745964 TI - Calcium-stimulated adenylyl cyclases modulate ethanol-induced neurodegeneration in the neonatal brain. AB - Fetal alcohol exposure results in cognitive and neurobehavioral deficits, but the effects of modifying genetic loci on the severity of these sequelas have not been well characterized. Although the cAMP signaling pathway has been shown to be an important modulator of ethanol sensitivity in adult mice, its potential role in modulating ethanol-induced neurodegeneration has not been examined. Adenylyl cyclases (ACs) 1 and 8 produce cAMP in response to intracellular calcium elevation and modulate several aspects of neuronal function, including ethanol sensitivity. AC1 and AC8 are expressed widely throughout the brain of neonatal mice, and genetic deletion of both AC1 and AC8 in double-knock-out (DKO) mice enhances ethanol-induced neurodegeneration in the brains of neonatal mice. In addition, ethanol treatment induces significantly greater levels of caspase-3 activation in the brains of DKO mice compared with wild-type (WT) mice, reflecting higher numbers of apoptotic neurons. Administration of the NMDA receptor antagonist MK801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine hydrogen maleate] or the GABA(A) receptor potentiator phenobarbital, which mimics components of the effects of ethanol on neurons, results in significantly greater neurodegeneration in the brains of neonatal DKO mice than WT mice. Furthermore, loss of a single calcium-stimulated AC isoform potentiates neurodegeneration after administration of ethanol, MK801, or phenobarbital. In contrast, the levels of physiological cell death, death after hypoxia/ischemia, and excitotoxic cell death are not increased in the brains of DKO mice. Thus, AC1 and AC8 are critical modulators of neurodegeneration induced by activity blockade in the neonatal brain and represent genetic loci that may potentially modify the severity of fetal alcohol syndrome. PMID- 15745963 TI - Endothelial nitric oxide synthase regulates brain-derived neurotrophic factor expression and neurogenesis after stroke in mice. AB - Here, we investigate the effects of endothelial nitric oxide synthase (eNOS) on angiogenesis, neurogenesis, neurotrophic factor expression, and neurological functional outcome after stroke. Wild-type and eNOS knock-out (eNOS-/-) mice were subjected to permanent occlusion of the right middle cerebral artery. eNOS-/- mice exhibited more severe neurological functional deficit after stroke than wild type mice. Decreased subventricular zone (SVZ) progenitor cell proliferation and migration, measured using bromodeoxyuridine, Ki-67, nestin, and doublecortin immunostaining in the ischemic brain, and decreased angiogenesis, as demonstrated by reduced endothelial cell proliferation, vessel perimeter, and vascular density in the ischemic border, were evident in eNOS-/- mice compared with wild-type mice. eNOS-deficient mice also exhibited a reduced response to vascular endothelial growth factor (VEGF)-induced angiogenesis in a corneal assay. ELISAs showed that eNOS-/- mice have decreased brain-derived neurotrophic factor (BDNF) expression but not VEGF and basic fibroblast growth factor in the ischemic brain compared with wild-type mice. In addition, cultured SVZ neurosphere formation, proliferation, telomerase activity, and neurite outgrowth but not cell viability from eNOS-/- mice were significantly reduced compared with wild-type mice. BDNF treatment of SVZ cells derived from eNOS-/- mice restored the decreased neurosphere formation, proliferation, neurite outgrowth, and telomerase activity in cultured eNOS(-/-) SVZ neurospheres. SVZ explant cell migration also was significantly decreased in eNOS-/- mice compared with wild-type mice. These data indicate that eNOS is not only a downstream mediator for VEGF and angiogenesis but also regulates BDNF expression in the ischemic brain and influences progenitor cell proliferation, neuronal migration, and neurite outgrowth and affects functional recovery after stroke. PMID- 15745965 TI - Axonal transport, amyloid precursor protein, kinesin-1, and the processing apparatus: revisited. AB - The sequential enzymatic actions of beta-APP cleaving enzyme 1 (BACE1), presenilins (PS), and other proteins of the gamma-secretase complex liberate beta amyloid (Abeta) peptides from larger integral membrane proteins, termed beta amyloid precursor proteins (APPs). Relatively little is known about the normal function(s) of APP or the neuronal compartment(s) in which APP undergoes proteolytic processing. Recent studies have been interpreted as consistent with the idea that APP serves as a kinesin-1 cargo receptor and that PS and BACE1 are associated with the APP-resident membranous cargos that undergo rapid axonal transport. In this report, derived from a collaboration among several independent laboratories, we examined the potential associations of APP and kinesin-1 using glutathione S-transferase pull-down and coimmunoprecipitation assays. In addition, we assessed the trafficking of membrane proteins in the sciatic nerves of transgenic mice with heterozygous or homozygous deletions of APP. In contrast to previous reports, we were unable to find evidence for direct interactions between APP and kinesin-1. Furthermore, the transport of kinesin-1 and tyrosine kinase receptors, previously reported to require APP, was unchanged in axons of APP-deficient mice. Finally, we show that two components of the APP proteolytic machinery, i.e., PS1 and BACE1, are not cotransported with APP in the sciatic nerves of mice. These findings suggest that the hypothesis that APP serves as a kinesin-1 receptor and that the proteolytic processing machinery responsible for generating Abeta is transported in the same vesicular compartment in axons of peripheral nerves requires revision. PMID- 15745966 TI - Cytoprotective effects of geranylgeranylacetone against retinal photooxidative damage. AB - Exposure to excessive light induces retinal photoreceptor cell damage, leading to development and progression of various retinal diseases. We tested the effect of geranylgeranylacetone (GGA), an acyclic polyisoprenoid, on light-induced retinal damage in mice. Oral treatment with GGA (1.0 mg/d) for 5 d induced thioredoxin (Trx) and heat shock protein 72 (Hsp72) predominantly in the retinal pigment epithelium (RPE). After white light exposure (8000 lux for 2 h), the percentage of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive photoreceptor cells decreased significantly at 24 and 96 h, and the number of photoreceptor cell nuclei at 96 h and the electroretinographic amplitudes of the a- and b-waves at 4 and 10 d increased significantly in GGA pretreated mice compared with saline-pretreated mice. Light-induced upregulations of 8-hydroxy-2-deoxyguanosine and 4-hydroxy-2-nonenal-modified protein, markers of oxidative stress, were inhibited by GGA pretreatment. To elucidate the cytoprotective mechanism of GGA and Trx, we used human K-1034 RPE cells and mouse photoreceptor-derived 661W cells. In K-1034 cells, GGA (10 microM) induced intracellular Trx, Hsp72, and extracellular Trx but not extracellular Hsp72. Extracellular Trx (0.75 nM) attenuated H2O2 (200 microM)-induced cell damage in 661W cells. Pretreatment with GGA and overexpression of Trx in K-1034 cells counteracted H2O2 (50 microM)-induced attenuation of cellular latex bead incorporation. Protection of phagocytotic activity through induction of Trx and possibly Hsp72 in RPE cells and elimination of oxidative stress in the photoreceptor layer through release of Trx from RPE cells may be mechanisms of GGA-mediated cytoprotection. Therefore, Trx is a neurotrophic factor released from RPE cells and plays a crucial role in maintaining photoreceptor cell integrity. PMID- 15745967 TI - Reduced nerve injury-induced neuropathic pain in kinin B1 receptor knock-out mice. AB - Injury to peripheral nerves often results in a persistent neuropathic pain condition that is characterized by spontaneous pain, allodynia, and hyperalgesia. Nerve injury is accompanied by a local inflammatory reaction in which nerve associated and immune cells release several pronociceptive mediators. Kinin B1 receptors are rarely expressed in nontraumatized tissues, but they can be expressed after tissue injury. Because B1 receptors mediate chronic inflammatory painful processes, we studied their participation in neuropathic pain using receptor gene-deleted mice. In the absence of neuropathy, we found no difference in the paw-withdrawal responses to thermal or mechanical stimulation between B1 receptor knock-out mice and 129/J wild-type mice. Partial ligation of the sciatic nerve in the wild-type mouse produced a profound and long-lasting decrease in thermal and mechanical thresholds in the paw ipsilateral to nerve lesion. Threshold changed neither in the sham-operated animals nor in the paw contralateral to lesion. Ablation of the gene for the B1 receptor resulted in a significant reduction in early stages of mechanical allodynia and thermal hyperalgesia. Furthermore, systemic treatment with the B1 selective receptor antagonist des-Arg9-[Leu8]-bradykinin reduced the established mechanical allodynia observed 7-28 d after nerve lesion in wild-type mice. Partial sciatic nerve ligation induced an upregulation in B1 receptor mRNA in ipsilateral paw, sciatic nerve, and spinal cord of wild-type mice. Together, kinin B1 receptor activation seems to be essential to neuropathic pain development, suggesting that an oral-selective B1 receptor antagonist might have therapeutic potential in the management of chronic pain. PMID- 15745968 TI - Beyond memory, navigation, and inhibition: behavioral evidence for hippocampus dependent cognitive coordination in the rat. AB - Injecting tetrodotoxin (TTX) into one hippocampus impaired avoidance of a place defined by distal cues while rats were on a slowly rotating arena. The impairment could be explained by a deficit in memory, navigation, or behavioral inhibition. Here, we show that the TTX injection abolished the ability of rats to organize place-avoidance behavior specifically when distal room and local arena cues were continuously dissociated. The results provide evidence that injecting TTX into one hippocampus specifically impaired the coordination of representations that support organized behavior because of the following: (1) rats normally coordinate separate room and arena avoidance memories; (2) the TTX injection spared spatial, relational, and representational memory, navigation, and behavioral inhibition; and (3) the TTX-induced impairment of place avoidance depended on the need to coordinate representations of local and distal stimuli. PMID- 15745969 TI - Degradation of head direction cell activity during inverted locomotion. AB - Head direction (HD) cells in the rat limbic system carry information about the direction the head is pointing in the horizontal plane. Most previous studies of HD functioning have used animals locomoting in an upright position or ascending/descending a vertical wall. In the present study, we recorded HD cell activity from the anterodorsal thalamic nucleus while the animal was locomoting in an upside-down orientation. Rats performed a shuttle-box task requiring them to climb a vertical wall and locomote across the ceiling of the apparatus while inverted to reach an adjoining wall before ascending into the reward compartment. The apparatus was oriented toward the preferred direction of the recorded cell, or the 180 degrees opposite direction. When the animal was traversing the vertical walls of the apparatus, the HD cells remained directionally tuned as if the walls were an extension of the floor. When the animal was locomoting inverted on the ceiling, however, cells showed a dramatic change in activity. Nearly one half (47%) of the recorded cells exhibited no directional specificity during inverted locomotion, despite showing robust directional tuning on the walls before and after inversion. The remaining cells showed significantly degraded measures of directional tuning and random shifts of the preferred direction relative to the floor condition while the animal was inverted. It has previously been suggested that the HD system uses head angular velocity signals from the vestibular system to maintain a consistent representation of allocentric direction. These findings suggest that being in an inverted position causes a distortion of the vestibular signal controlling the HD system. PMID- 15745970 TI - Physiological changes in glucose differentially modulate the excitability of hypothalamic melanin-concentrating hormone and orexin neurons in situ. AB - The physiological signaling mechanisms that link normal variations in body energy status to the activity of arousal- and metabolism-regulating brain centers are not well understood. The melanin-concentrating hormone (MCH) and orexin/hypocretin types of neurons of the lateral hypothalamus (LH) exert opposing effects on arousal and metabolism. We examined whether shifts in brain extracellular glucose that correspond to physiological changes in blood glucose can alter the electrical output of neurochemically and biophysically defined LH cells in mouse brain slices. Here, we show that physiologically relevant concentrations of glucose dose-dependently enhance the electrical excitability of MCH neurons by inducing depolarization and increasing membrane resistance. We also demonstrate that the same physiological shifts in glucose have the opposite effects on the electrical activity of orexin neurons. We propose that these direct actions of glucose on the arousal- and metabolism-regulating LH neurons play a key role in the translation of normal variations in body energy resources into appropriate changes in arousal and metabolism. PMID- 15745972 TI - Images in clinical medicine. Tuberculosis empyema necessitatis. PMID- 15745971 TI - Coxsackievirus targets proliferating neuronal progenitor cells in the neonatal CNS. AB - Type B coxsackieviruses (CVB) frequently infect the CNS and, together with other enteroviruses, are the most common cause of viral meningitis in humans. Newborn infants are particularly vulnerable, and CVB also can infect the fetus, leading to mortality, or to neurodevelopmental defects in surviving infants. Using a mouse model of neonatal CVB infection, we previously demonstrated that coxsackievirus B3 (CVB3) could infect neuronal progenitor cells in the subventricular zone (SVZ). Here we extend these findings, and we show that CVB3 targets actively proliferating (bromodeoxyuridine+, Ki67+) cells in the SVZ, including type B and type A stem cells. However, infected cells exiting the SVZ have lost their proliferative capacity, in contrast to their uninfected companions. Despite being proliferation deficient, the infected neuronal precursors could migrate along the rostral migratory stream and radial glia, to reach their final destinations in the olfactory bulb or cerebral cortex. Furthermore, infection did not prevent cell differentiation, as determined by cellular morphology and the expression of maturation markers. These data lead us to propose a model of CVB infection of the developing CNS, which may explain the neurodevelopmental defects that result from fetal infection. PMID- 15745973 TI - Overbilling vs. downcoding--the battle between physicians and insurers. PMID- 15745974 TI - Turf battles in coronary revascularization. PMID- 15745975 TI - Personal digital educators. PMID- 15745976 TI - Genetic factors in Alzheimer's disease. PMID- 15745977 TI - U.K. Controlled trial of intrapleural streptokinase for pleural infection. AB - BACKGROUND: Intrapleural fibrinolytic agents are used in the drainage of infected pleural-fluid collections. This use is based on small trials that did not have the statistical power to evaluate accurately important clinical outcomes, including safety. We conducted a trial to clarify the therapeutic role of intrapleural streptokinase. METHODS: In this double-blind trial, 454 patients with pleural infection (defined by the presence of purulent pleural fluid or pleural fluid with a pH below 7.2 with signs of infection or by proven bacterial invasion of the pleural space) were randomly assigned to receive either intrapleural streptokinase (250,000 IU twice daily for three days) or placebo. Patients received antibiotics and underwent chest-tube drainage, surgery, and other treatment as part of routine care. The number of patients in the two groups who had died or needed surgical drainage at three months was compared (the primary end point); secondary end points were the rates of death and of surgery (analyzed separately), the radiographic outcome, and the length of the hospital stay. RESULTS: The groups were well matched at baseline. Among the 427 patients who received streptokinase or placebo, there was no significant difference between the groups in the proportion of patients who died or needed surgery (with streptokinase: 64 of 206 patients [31 percent]; with placebo: 60 of 221 [27 percent]; relative risk, 1.14 [95 percent confidence interval, 0.85 to 1.54; P=0.43), a result that excluded a clinically significant benefit of streptokinase. There was no benefit to streptokinase in terms of mortality, rate of surgery, radiographic outcomes, or length of the hospital stay. Serious adverse events (chest pain, fever, or allergy) were more common with streptokinase (7 percent, vs. 3 percent with placebo; relative risk, 2.49 [95 percent confidence interval, 0.98 to 6.36]; P=0.08). CONCLUSIONS: The intrapleural administration of streptokinase does not improve mortality, the rate of surgery, or the length of the hospital stay among patients with pleural infection. PMID- 15745978 TI - Quantitative determinants of the outcome of asymptomatic mitral regurgitation. AB - BACKGROUND: The clinical outcome of asymptomatic mitral regurgitation is poorly defined, and the treatment is uncertain. We studied the effect on the outcome of quantifying mitral regurgitation according to recent guidelines. METHODS: We prospectively enrolled 456 patients (mean [+/-SD] age, 63+/-14 years; 63 percent men; ejection fraction, 70+/-8 percent) with asymptomatic organic mitral regurgitation, quantified according to current recommendations (regurgitant volume, 66+/-40 ml per beat; effective regurgitant orifice, 40+/-27 mm2). RESULTS: The estimated five-year rates (+/-SE) of death from any cause, death from cardiac causes, and cardiac events (death from cardiac causes, heart failure, or new atrial fibrillation) with medical management were 22+/-3 percent, 14+/-3 percent, and 33+/-3 percent, respectively. Independent determinants of survival were increasing age, the presence of diabetes, and increasing effective regurgitant orifice (adjusted risk ratio per 10-mm2 increment, 1.18; 95 percent confidence interval, 1.06 to 1.30; P<0.01), the predictive power of which superseded all other qualitative and quantitative measures of regurgitation. Patients with an effective regurgitant orifice of at least 40 mm2 had a five-year survival rate that was lower than expected on the basis of U.S. Census data (58+/ 9 percent vs. 78 percent, P=0.03). As compared with patients with a regurgitant orifice of less than 20 mm2, those with an orifice of at least 40 mm2 had an increased risk of death from any cause (adjusted risk ratio, 2.90; 95 percent confidence interval, 1.33 to 6.32; P<0.01), death from cardiac causes (adjusted risk ratio, 5.21; 95 percent confidence interval, 1.98 to 14.40; P<0.01), and cardiac events (adjusted risk ratio, 5.66; 95 percent confidence interval, 3.07 to 10.56; P<0.01). Cardiac surgery was ultimately performed in 232 patients and was independently associated with improved survival (adjusted risk ratio, 0.28; 95 percent confidence interval, 0.14 to 0.55; P<0.01). CONCLUSIONS: Quantitative grading of mitral regurgitation is a powerful predictor of the clinical outcome of asymptomatic mitral regurgitation. Patients with an effective regurgitant orifice of at least 40 mm2 should promptly be considered for cardiac surgery. PMID- 15745979 TI - Family-based association between Alzheimer's disease and variants in UBQLN1. AB - BACKGROUND: Recent analyses suggest that the known Alzheimer's disease genes account for less than half the genetic variance in this disease. The gene encoding ubiquilin 1 (UBQLN1) is one of several candidate genes for Alzheimer's disease located near a well-established linkage peak on chromosome 9q22. METHODS: We evaluated 19 single-nucleotide polymorphisms in three genes within the chromosome 9q linkage region in 437 multiplex families with Alzheimer's disease from the National Institute of Mental Health (NIMH) sample (1439 subjects). We then tested the single-nucleotide polymorphisms showing a positive result in an independently identified set of 217 sibships discordant for Alzheimer's disease (Consortium on Alzheimer's Genetics [CAG] sample; 489 subjects) and assessed the functional effect of an implicated single-nucleotide polymorphism in brain tissue from 25 patients with Alzheimer's disease and 17 controls. RESULTS: In the NIMH sample, we observed a significant association between Alzheimer's disease and various single-nucleotide polymorphisms in UBQLN1. We confirmed these associations in the CAG sample. The risk-conferring haplotype in both samples was defined by a single intronic single-nucleotide polymorphism located downstream of exon 8. The risk allele was associated with a dose-dependent increase in an alternatively spliced UBQLN1 (lacking exon 8) transcript in RNA extracted from brain samples of patients with Alzheimer's disease. CONCLUSIONS: Our findings suggest that genetic variants in UBQLN1 on chromosome 9q22 substantially increase the risk of Alzheimer's disease, possibly by influencing alternative splicing of this gene in the brain. PMID- 15745980 TI - Risks and benefits of phase 1 oncology trials, 1991 through 2002. AB - BACKGROUND: Previous reviews of phase 1 oncology trials reported a rate of response to treatment of 4 to 6 percent and a toxicity-related death rate of 0.5 percent. These results may not reflect the rates in current phase 1 oncology trials. METHODS: We reviewed all nonpediatric phase 1 oncology trials sponsored by the Cancer Therapy Evaluation Program at the National Cancer Institute between 1991 and 2002. We report the rates of response to treatment, of stable disease, of grade 4 toxic events, and of treatment-related deaths. RESULTS: We analyzed 460 trials involving 11,935 participants, all of whom were assessed for toxicity and 10,402 of whom were assessed for a response to therapy. The overall response rate (i.e., for both complete and partial responses) was 10.6 percent, with considerable variation among trials. "Classic" phase 1 trials of single investigational chemotherapeutic agents represented only 20 percent of the trials and had a response rate of 4.4 percent. Studies that included at least one anticancer agent approved by the Food and Drug Administration constituted 46.3 percent of the trials and had a response rate of 17.8. An additional 34.1 percent of participants had stable disease or a less-than-partial response. The overall rate of death due to toxic events was 0.49 percent. Of 3465 participants for whom data on patient-specific grade 4 toxic events were available, 14.3 percent had had at least one episode of grade 4 toxic events. CONCLUSIONS: Overall response rates among phase 1 oncology trials are higher than previously reported, although they have not changed for classic phase 1 trials, and toxicity-related death rates have remained stable. Rates of response and toxicity vary, however, among the various types of phase 1 oncology trials. PMID- 15745981 TI - Antithyroid drugs. PMID- 15745982 TI - Images in clinical medicine. Dermopathy of Graves' disease. PMID- 15745983 TI - Clinical problem-solving. On the threshold--a diagnosis of exclusion. PMID- 15745984 TI - Multicenter trials of treatment for empyema--after all these years. PMID- 15745985 TI - Timing of surgery in asymptomatic mitral regurgitation. PMID- 15745986 TI - Risks and benefits of phase 1 oncology trials, revisited. PMID- 15745987 TI - Toward a new therapy for tuberculosis. PMID- 15745988 TI - Umbilical-cord blood for transplantation in adults. PMID- 15745989 TI - ACE inhibition in stable coronary artery disease. PMID- 15745990 TI - Premature birth and insulin resistance. PMID- 15745991 TI - Radial-artery coronary bypass grafts. PMID- 15745992 TI - Pharmacists and emergency contraception. PMID- 15745993 TI - Lymphangitis after self-administration of lipopolysaccharide. PMID- 15745994 TI - More on Phineas Gage. PMID- 15745995 TI - Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist. AB - We have developed an automated SELEX (Systematic Evolution of Ligands by EXponential Enrichment) process that allows the execution of in vitro selection cycles without any direct manual intervention steps. The automated selection protocol is designed to provide for high flexibility and versatility in terms of choice of buffers and reagents as well as stringency of selection conditions. Employing the automated SELEX process, we have identified RNA aptamers to the mirror-image configuration (d-peptide) of substance P. The peptide substance P belongs to the tachykinin family and exerts various biologically important functions, such as peripheral vasodilation, smooth muscle contraction and pain transmission. The aptamer that was identified most frequently was truncated to the 44mer SUP-A-004. The mirror-image configuration of SUP-A-004, the so-called Spiegelmer, has been shown to bind to naturally occurring l-substance P displaying a K(d) of 40 nM and to inhibit (IC50 of 45 nM) l-substance P-mediated Ca2+ release in a cell culture assay. PMID- 15745996 TI - Amino-functionalized DNA: the properties of C5-amino-alkyl substituted 2' deoxyuridines and their application in DNA triplex formation. AB - The incorporation of C5-amino-modified 2'-deoxyuridine analogues into DNA have found application in nucleic acid labelling, the stabilization of nucleic acid structures, functionalization of nucleic acid aptamers and catalysts, and the investigation of sequence-specific DNA bending. In this study, we describe the physicochemical properties of four different C5-amino-modified 2'-deoxyuridines in which the amino group is tethered to the base via a 3-carbon alkyl, Z- or E alkenyl or alkynyl linker. Conformational parameters of the nucleosides and their pK(a) values were deduced using 1H NMR. All of them display the expected anti conformation of the nucleoside with 2'-endo sugar puckers for the deoxyribose ring. A preference for the cisoid conformation for the Z-alkenyl analogue is found, while the E-alkenyl analogue exists exclusively as its transoid conformation. The pK(a) values range from 10.0 for the analogue with an aliphatic propyl linker to 8.5 for the propargylamino analogue. The analogues have been used for the synthesis of triple-helix forming oligonucleotides (TFOs) in which they replace thymidine in the natural sequence. Oligonucleotides containing the propargylamino analogue display the highest stability especially at low pH, while those containing analogues with propyl and especially Z-alkenyl linkers are destabilized to a great extent. TFOs containing the analogue with the E-alkenyl linker have stability similar to the unmodified structures. The chemical synthesis of TFOs containing the analogue, 5-(3-hydroxyprop-1-ynyl)-2' deoxyuridine that possesses a neutral but polar side chain show a remarkable stability, which is higher than that of all TFOs containing the alkylamino or alkenylamino analogues and only slightly lower than that of TFOs containing the propargylamino analogue. Both the hydroxyl and propargylamino substitutions impart enhanced triple-helix stability relative to the analogous sequences containing C5-propynyl-2'-deoxyuridine. Furthermore, a similar dependence of stability on pH is found between TFOs containing the hydroxypropynyl modifications and those containing the propargylamino side chains. This suggests that the major factor responsible for stabilizing such triple helices is due to the presence of the alkyne with an attached electronegative group. PMID- 15745997 TI - Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/helicase and a species-specific interaction between RPA and Dna2. AB - In both budding and fission yeasts, a null mutation of the DNA2 gene is lethal. In contrast, a null mutation of Caenorhabditis elegans dna2+ causes a delayed lethality, allowing survival of some mutant C.elegans adults to F2 generation. In order to understand reasons for this difference in requirement of Dna2 between these organisms, we examined the enzymatic properties of the recombinant C.elegans Dna2 (CeDna2) and its interaction with replication-protein A (RPA) from various sources. Like budding yeast Dna2, CeDna2 possesses DNA-dependent ATPase, helicase and endonuclease activities. The specific activities of both ATPase and endonuclease activities of the CeDna2 were considerably higher than the yeast Dna2 (approximately 10- and 20-fold, respectively). CeDna2 endonuclease efficiently degraded a short 5' single-stranded DNA tail (<10 nt) that was hardly cleaved by ScDna2. Both endonuclease and helicase activities of CeDna2 were stimulated by CeRPA, but not by human or yeast RPA, demonstrating a species specific interaction between Dna2 and RPA. These and other enzymatic properties of CeDna2 described in this paper may shed light on the observation that C.elegans is less stringently dependent on Dna2 for its viability than Saccharomyces cerevisiae. We propose that flaps generated by DNA polymerase delta mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability. PMID- 15745998 TI - Modular RNA architecture revealed by computational analysis of existing pseudoknots and ribosomal RNAs. AB - Modular architecture is a hallmark of RNA structures, implying structural, and possibly functional, similarity among existing RNAs. To systematically delineate the existence of smaller topologies within larger structures, we develop and apply an efficient RNA secondary structure comparison algorithm using a newly developed two-dimensional RNA graphical representation. Our survey of similarity among 14 pseudoknots and subtopologies within ribosomal RNAs (rRNAs) uncovers eight pairs of structurally related pseudoknots with non-random sequence matches and reveals modular units in rRNAs. Significantly, three structurally related pseudoknot pairs have functional similarities not previously known: one pair involves the 3' end of brome mosaic virus genomic RNA (PKB134) and the alternative hammerhead ribozyme pseudoknot (PKB173), both of which are replicase templates for viral RNA replication; the second pair involves structural elements for translation initiation and ribosome recruitment found in the viral internal ribosome entry site (PKB223) and the V4 domain of 18S rRNA (PKB205); the third pair involves 18S rRNA (PKB205) and viral tRNA-like pseudoknot (PKB134), which probably recruits ribosomes via structural mimicry and base complementarity. Additionally, we quantify the modularity of 16S and 23S rRNAs by showing that RNA motifs can be constructed from at least 210 building blocks. Interestingly, we find that the 5S rRNA and two tree modules within 16S and 23S rRNAs have similar topologies and tertiary shapes. These modules can be applied to design novel RNA motifs via build-up-like procedures for constructing sequences and folds. PMID- 15745999 TI - metaSHARK: software for automated metabolic network prediction from DNA sequence and its application to the genomes of Plasmodium falciparum and Eimeria tenella. AB - The metabolic SearcH And Reconstruction Kit (metaSHARK) is a new fully automated software package for the detection of enzyme-encoding genes within unannotated genome data and their visualization in the context of the surrounding metabolic network. The gene detection package (SHARKhunt) runs on a Linux system and requires only a set of raw DNA sequences (genomic, expressed sequence tag and/or genome survey sequence) as input. Its output may be uploaded to our web-based visualization tool (SHARKview) for exploring and comparing data from different organisms. We first demonstrate the utility of the software by comparing its results for the raw Plasmodium falciparum genome with the manual annotations available at the PlasmoDB and PlasmoCyc websites. We then apply SHARKhunt to the unannotated genome sequences of the coccidian parasite Eimeria tenella and observe that, at an E-value cut-off of 10(-20), our software makes 142 additional assertions of enzymatic function compared with a recent annotation package working with translated open reading frame sequences. The ability of the software to cope with low levels of sequence coverage is investigated by analyzing assemblies of the E.tenella genome at estimated coverages from 0.5x to 7.5x. Lastly, as an example of how metaSHARK can be used to evaluate the genomic evidence for specific metabolic pathways, we present a study of coenzyme A biosynthesis in P.falciparum and E.tenella. PMID- 15746001 TI - Functional anatomy of biological motion perception in posterior temporal cortex: an FMRI study of eye, mouth and hand movements. AB - Passive viewing of biological motion engages extensive regions of the posterior temporal-occipital cortex in humans, particularly within and nearby the superior temporal sulcus (STS). Relatively little is known about the functional specificity of this area. Some recent studies have emphasized the perceived intentionality of the motion as a potential organizing principle, while others have suggested the existence of a somatotopy based upon the limb perceived in motion. Here we conducted an event-related functional magnetic resonance imaging experiment to compare activity elicited by movement of the eyes, mouth or hand. Each motion evoked robust activation in the right posterior temporal-occipital cortex. While there was substantial overlap of the activation maps in this region, the spatial distribution of hemodynamic response amplitudes differentiated the movements. Mouth movements elicited activity along the mid posterior STS while eye movements elicited activity in more superior and posterior portions of the right posterior STS region. Hand movements activated more inferior and posterior portions of the STS region within the posterior continuing branch of the STS. Hand-evoked activity also extended into the inferior temporal, middle occipital and lingual gyri. This topography may, in part, reflect the role of particular body motions in different functional activities. PMID- 15746000 TI - Cadmium inhibits mismatch repair by blocking the ATPase activity of the MSH2-MSH6 complex. AB - Cadmium (Cd2+) is a known carcinogen that inactivates the DNA mismatch repair (MMR) pathway. In this study, we have tested the effect of Cd2+ exposure on the enzymatic activity of the mismatch binding complex MSH2-MSH6. Our results indicate that Cd2+ is highly inhibitory to the ATP binding and hydrolysis activities of MSH2-MSH6, and less inhibitory to its DNA mismatch binding activity. The inhibition of the ATPase activity appears to be dose and exposure time dependent. However, the inhibition of the ATPase activity by Cd2+ is prevented by cysteine and histidine, suggesting that these residues are essential for the ATPase activity and are targeted by Cd2+. A comparison of the mechanism of inhibition with N-ethyl maleimide, a sulfhydryl group inhibitor, indicates that this inhibition does not occur through direct inactivation of sulfhydryl groups. Zinc (Zn2+) does not overcome the direct inhibitory effect of Cd2+ on the MSH2-MSH6 ATPase activity in vitro. However, the increase in the mutator phenotype of yeast cells exposed to Cd2+ was prevented by excess Zn2+, probably by blocking the entry of Cd2+ into the cell. We conclude that the inhibition of MMR by Cd2+ is through the inactivation of the ATPase activity of the MSH2-MSH6 heterodimer, resulting in a dominant negative effect and causing a mutator phenotype. PMID- 15746002 TI - Monetary incentives enhance processing in brain regions mediating top-down control of attention. AB - To evaluate the effect of an abstract motivational incentive on top-down mechanisms of visual spatial attention, 10 subjects engaged in a target detection task and responded to targets preceded by spatially valid (predictive), invalid (misleading) or neutral central cues under three different incentive conditions: win money (WIN), lose money (LOSE), and neutral (neither gain nor lose). Activation in the posterior cingulate cortex was correlated with visual spatial expectancy, defined as the degree to which the valid cue benefited performance as evidenced by faster reaction times compared to non-directional cues. Winning and losing money enhanced this relationship via overlapping but independent limbic mechanisms. In addition, activity in the inferior parietal lobule was correlated with disengagement (the degree to which invalid cues diminished performance). This relationship was also enhanced by monetary incentives. Finally, incentive enhanced the relationship of activation in the visual cortex to visual spatial expectancy and disengagement for both types of incentive (WIN and LOSE). These results show that abstract incentives enhance neural processing within the attention network in a process- and valence-selective manner. They also show that different cognitive and motivational mechanisms may produce a common effect upon unimodal cortices in order to enhance processing to serve the current behavioral goal. PMID- 15746003 TI - Functional neuroanatomy of human vocalization: an H215O PET study. AB - Vocalization in lower animals is associated with a well-described visceromotor call system centered on the mesencephalic periacqueductal grey matter (PAG), which is itself regulated by paramedian cortical structures. To determine the role this phylogenetically older system plays in human phonation, we contrasted voiced and unvoiced speech using positron emission tomography and then evaluated functional connectivity of regions that significantly differentiated these conditions. Vocalization was associated with increased and highly correlated activity within the midline structures--PAG and paramedian cortices--described in lower mammalian species. Concurrent activation and connectivity of neocortical and subcortical motor regions--medial and lateral premotor structures and elements of basal ganglia thalamocortical circuitry--suggest a mechanism by which this system may have come under an increasing degree of voluntary control in humans. Additionally, areas in the temporal lobe and cerebellum were selectively activated during voiced but not unvoiced speech. These regions are functionally coupled to both visceromotor and neocortical motor areas during production of voiced speech, suggesting they may play a central role in self-monitoring and feedback regulation of human phonation. PMID- 15746004 TI - Impairment of gaze-centered updating of reach targets in bilateral parietal occipital damaged patients. AB - Recent studies have suggested that internal updating of visuospatial targets in humans occurs in gaze-centered coordinates and takes place in the parietal and extrastriate cortices. We explored how information for reaching is updated in two patients with bilateral lesions in these areas. Subjects performed two visuomotor tasks: (i) a fixation reaching task, which began with the appearance of one of five fixation positions (varying eye positions) followed by a central reaching target. Subjects reached to the target while fixating on the presented fixation position (relative to gaze the target was always presented in the periphery); and (ii) a saccade reaching task, in which subjects foveated on the central reaching target, then made a saccade to the presented fixation position before reaching to the central target. In both tasks, subjects reached to targets after a 500 or 5000 ms delay. Gaze-centered updating predicts similarities in reaching errors between fixation and saccade trials. Control subjects showed evidence for gaze centered updating during both 500 and 5000 ms delay conditions. In contrast, patient AT, who had extensive occipital-parietal damage, only showed signs of gaze-centered representation after 5 s. Patient IG, with a more focal lesion in the parietal cortices, showed partial updating in gaze-centered coordinates when reaching with the small memory delay but recovered a complete gaze-centered representation after the longer delay. This suggests that patients with bilateral occipital-parietal lesions may rely on non-gaze-centered frames to store immediate target locations in reaching space but, given enough time, this information may be rerouted to access other gaze-centered motor cortical mechanisms. PMID- 15746005 TI - CO inhalation at dose corresponding to tobacco smoke worsens cardiac remodeling after experimental myocardial infarction in rats. AB - We hypothesized that inhalation of carbon monoxide (CO) (500 ppm), similar to that in tobacco smoke, disturbs the cardiovascular adaptation after myocardial infarction by increasing remodeling. Four groups of rats were assessed. Two groups had myocardial infarction induced by the ligation of the left coronary artery: the first group was exposed to air (infarcted air group, n = 12), and the second was exposed to CO (infarcted CO group, n = 11). They were compared to two sham-operated groups, a control air group (n = 10), and a control CO group (n = 7) exposed (3 weeks) to CO. Aerobic endurance capacity was assessed in both the infarct CO and infarct air group (endurance capacity = 0.043 +/- 0.006 m.min( 1).g(-1) vs. 0.042 +/- 0.005 m.min(-1).g(-1), not significant). In the infarcted CO group compared to the infarcted air group, the dilatation of the left ventricle observed 3 weeks after infarction was increased, (left ventricular diastolic (LVD) diameter (D) = 9 +/- 0.4 vs. 7 +/- 0.4 mm, p < 0.05; left ventricular systolic (LVS) diameter (D) = 6 +/- 0.6 vs. 4.1 +/- 0.4, p < 0.05), and the diastolic posterior wall thickness was augmented (posterior wall diastolic thickness = 1.7 +/- 0.1 vs. 1.3 +/- 0.1 mm, p < 0.05). Hemodynamic pressure measurements in both ventricles and pulmonary artery showed elevated diastolic pressure after CO exposure compared to air exposure (LVD pressure = 32 +/- 1.6 vs. 19 +/- 2.3 mm Hg, p < 0.05; right ventricular diastolic pressure = 16 +/- 1.6 vs. 8.6 +/- 1.6 mm Hg, p < 0.05; pulmonary arterial pressure in diastole (PAD) = 27 +/- 1.6 vs. 20 +/- 2.3 mm Hg, p < 0.05). In the infarcted CO group, the infarct size increased. Echocardiography and histology showed hypertrophy of the contralateral wall similar to that observed in the noninfarcted control CO group. In conclusion, chronic CO inhalation worsens heart failure in rats with myocardial infarction by an increase in the infarct size and hypertrophy remodeling. PMID- 15746006 TI - Identification of a rapid detoxification mechanism for brevetoxin in rats. AB - We examined detoxification of brevetoxin in rats through metabolic activities and key elimination routes by analyzing samples from individual rats exposed to two brevetoxin congeners (PbTx-2 and PbTx-3). Brevetoxins were detected by radioimmunoassay in methanolic extracts of blood within 1 h post intraperitoneal (ip) administration. The toxin assay response was about three times higher in PbTx-2-treated rats versus the same dose (180 microg/kg) of PbTx-3. This difference persisted for up to 8 h postexposure. When the blood samples were reextracted with 20% methanol to enhance recovery of potential polar brevetoxin metabolites, 25-fold higher assay activity was present in the PbTx-2-treated rats. Analysis of urine from the same animals identified 7-fold more activity in the PbTx-2-treated rats that accumulated over the course of 24 h. Radioimmunoassay-guided high performance liquid chromatographic analysis of urine from PbTx-2-treated rats yielded three major peaks of activity. The first peak was attributed to the two cysteine adducts, cysteine-PbTx sulfoxide and cysteine PbTx (MH(+): m/z 1034 and 1018). The second peak was attributed to the oxidized form of PbTx-2 (MH(+): m/z 911) and its reduction product PbTx-3. The third peak remains unidentified. Brevetoxin cysteine conjugate and its sulfoxide product contributed nearly three-quarters of the brevetoxin immunoactivity. Our findings indicate the most commonly occurring PbTx-2 is rapidly transformed to a polar metabolite of a reduced biological activity that appears in blood and remains for up to 8 h, yet is cleared mostly to the urine within 24 h. PMID- 15746007 TI - Acute cardiopulmonary alterations induced by fine particulate matter of Sao Paulo, Brazil. AB - The mechanisms involved in the association between air pollution and increased cardiovascular morbidity are not fully understood. The objective of this study was to test the hypothesis that fine particulate matter (PM(2.5)) induces systemic inflammation and vasoconstriction of small arteries in the lung and heart of rats. Thirty-eight healthy Wistar rats were anesthetized, intubated, and submitted to the instillation of 1 ml of distilled water diluted in the following solution: blank filter, 100 microg and 500 microg of PM(2.5). PM(2.5) was collected in glass fiber filters with a high-volume sampler. The animals were sacrificed 24 h after instillation when blood, heart, and lung samples were collected for morphological and wet-to-dry weight ratio analysis. PM(2.5) consisted of the following elements: sulphur, arsenic, bromine, chlorine, cobalt, iron, lanthanum, manganese, antimony, scandium, and thorium. Total reticulocytes significantly increased at both PM(2.5) doses (p < 0.05) while hematocrit levels increased in the 500 microg group (p < 0.05). Quantification of segmented neutrophils and fibrinogen levels showed a significant decrease, while lymphocytes counting increased with 100 microg of PM(2.5) (p < 0.05). A significant dose-dependent decrease of intra-acinar pulmonary arteriole lumen/wall ratio (L/W) was observed in PM groups (p < 0.001). Peribronchiolar arterioles L/W showed a significant decrease in the 500 microg group (p < 0.001). A significant increase in heart wet-to-dry weight ratio was observed in the 500 microg group (p < 0.001). In conclusion, fine environment particles in the city of Sao Paulo promote pulmonary and cardiac histological alterations. Pulmonary vasculature was markedly affected by particle instillation, resulting in significant vasoconstriction in healthy rats. PMID- 15746008 TI - Effects of in utero and lactational TCDD exposure on bone development in differentially sensitive rat lines. AB - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a notorious model compound of highly toxic environmental pollutants, polychlorinated dibenzo-p-dioxins (PCDDs). Their toxic effects are mediated via cytosolic aryl hydrocarbon receptor (AHR). We studied the effects of several dose levels of TCDD on developing rat bone after maternal exposure at different times of gestation and lactation in three differentially sensitive rat lines. Rat lines A, B, and C differ in their sensitivity to TCDD due to mutated AHR (Ahr(hw)) in line A and another TCDD resistance allele (B(hw)) in line B. Line C rats have no resistance alleles. Offspring were analyzed for bone mineral density and geometry by peripheral quantitative computed tomography (pQCT) and for bone biomechanics by three-point bending at mid-diaphysis of tibia and femur and by axial loading at femoral neck. TCDD treatment resulted in bone defects, mainly in offspring of the most sensitive line C at a maternal dose of 1 microg/kg. They included decreased bone length, cross-sectional area of cortex, and bone mineral density. Mechanical testing revealed significantly reduced bending breaking force and stiffness of tibia, femur, and femoral neck. The effects were exposure time-dependent, and earlier exposure caused more severe defects. Gestational exposure alone was not sufficient, but lactational exposure was required to cause the bone defects. Most of the defects were recovered at the age of 1 year. The results indicate that dioxins affect developing bone by interfering with bone growth and mechanical strength and that the effects are mainly reversible. The dioxin-resistance alleles, Ahr(hw) and B(hw) increase the resistance to these defects. PMID- 15746009 TI - Evaluation of oral and intravenous route pharmacokinetics, plasma protein binding, and uterine tissue dose metrics of bisphenol A: a physiologically based pharmacokinetic approach. AB - Bisphenol A (BPA) is a weakly estrogenic monomer used in the production of polycarbonate plastic and epoxy resins, both of which are used in food contact and other applications. A physiologically based pharmacokinetic (PBPK) model of BPA pharmacokinetics in rats and humans was developed to provide a physiological context in which the processes controlling BPA pharmacokinetics (e.g., plasma protein binding, enterohepatic recirculation of the glucuronide [BPAG]) could be incorporated. A uterine tissue compartment was included to allow the correlation of simulated estrogen receptor (ER) binding of BPA with increases in uterine wet weight (UWW) in rats. Intravenous- and oral-route blood kinetics of BPA in rats and oral-route plasma and urinary elimination kinetics in humans were well described by the model. Simulations of rat oral-route BPAG pharmacokinetics were less exact, most likely the result of oversimplification of the GI tract compartment. Comparison of metabolic clearance rates derived from fitting rat i.v. and oral-route data implied that intestinal glucuronidation of BPA is significant. In rats, but not humans, terminal elimination rates were strongly influenced by enterohepatic recirculation. In the absence of BPA binding to plasma proteins, simulations showed high ER occupancy at doses without uterine effects. Restricting free BPA to the measured unbound amount demonstrated the importance of including plasma binding in BPA kinetic models: the modeled relationship between ER occupancy and UWW increases was consistent with expectations for a receptor-mediated response with low ER occupancy at doses with no response and increasing occupancy with larger increases in UWW. PMID- 15746011 TI - Functional divergence and horizontal transfer of type IV secretion systems. AB - The type IV secretion system (TFSSs) is a multifunctional family of translocation pathways that mediate the transfer of DNA among bacteria and deliver DNA and proteins to eukaryotic cells during bacterial infections. Horizontal transmission has dominated the evolution of the TFSS, as demonstrated here by a lack of congruence between the tree topology inferred from components of the TFSS and the presumed bacterial species divergence pattern. A parsimony analysis suggests that conjugation represents the ancestral state and that the divergence from conjugation to secretion of effector molecules has occurred independently at multiple sites in the tree. The result shows that the nodes at which functional shifts have occurred coincide with those of horizontal gene transfers among distantly related bacteria. We suggest that it is the transfer between species that paved the way for the divergence of the TFSSs and discuss the general role of horizontal gene transfers for the evolution of novel gene functions. PMID- 15746010 TI - Manganese acts centrally to stimulate luteinizing hormone secretion: a potential influence on female pubertal development. AB - Manganese (Mn), an essential element considered important for normal growth and reproduction, has been shown in adults to be detrimental to reproductive function when elevated. Because Mn can cross the blood-brain barrier and accumulate in the hypothalamus, and because it has been suggested that infants and children are potentially more sensitive to Mn than adults, we wanted to determine the effects of Mn exposure on puberty-related hormones and the onset of female puberty. We demonstrated that MnCl(2) when administered acutely into the third ventricle of the brain acts dose-dependently to stimulate luteinizing hormone (LH) release in prepubertal female rats. Incubation of hypothalami in vitro showed that this effect was due to a Mn-induced stimulation of luteinizing hormone releasing hormone (LHRH). Further demonstration that this is a hypothalamic site of action was shown by in vivo blockade of LHRH receptors and lack of a direct pituitary action of Mn to stimulate LH in vitro. To assess potential short-term effects, animals were supplemented with MnCl(2) (10 mg/kg) by gastric gavage from day 12 until day 29, or, in other animals, until vaginal opening (VO). Mn caused elevated serum levels of LH, follicle stimulating hormone, and estradiol, and it initiated a moderate but significant advancement in age at VO. Our results are the first to show that Mn can stimulate specific puberty-related hormones and suggest that it may facilitate the normal onset of puberty. They also suggest that Mn may contribute to precocious puberty if an individual is exposed to elevated levels of Mn too early in development. PMID- 15746012 TI - Likelihood, parsimony, and heterogeneous evolution. AB - Evolutionary rates vary among sites and across the phylogenetic tree (heterotachy). A recent analysis suggested that parsimony can be better than standard likelihood at recovering the true tree given heterotachy. The authors recommended that results from parsimony, which they consider to be nonparametric, be reported alongside likelihood results. They also proposed a mixture model, which was inconsistent but better than either parsimony or standard likelihood under heterotachy. We show that their main conclusion is limited to a special case for the type of model they study. Their mixture model was inconsistent because it was incorrectly implemented. A useful nonparametric model should perform well over a wide range of possible evolutionary models, but parsimony does not have this property. Likelihood-based methods are therefore the best way to deal with heterotachy. PMID- 15746013 TI - Evolution of proteins and gene expression levels are coupled in Drosophila and are independently associated with mRNA abundance, protein length, and number of protein-protein interactions. AB - Organismic evolution requires that variation at distinct hierarchical levels and attributes be coherently integrated, often in the face of disparate environmental and genetic pressures. A central part of the evolutionary analysis of biological systems remains to decipher the causal connections between organism-wide (or genome-wide) attributes (e.g., mRNA abundance, protein length, codon bias, recombination rate, genomic position, mutation rate, etc) as well as their role together with mutation, selection, and genetic drift-in shaping patterns of evolutionary variation in any of the attributes themselves. Here we combine genome-wide evolutionary analysis of protein and gene expression data to highlight fundamental relationships among genomic attributes and their associations with the evolution of both protein sequences and gene expression levels. Our results show that protein divergence is positively coupled with both gene expression polymorphism and divergence. We show moreover that although the number of protein-protein interactions in Drosophila is negatively associated with protein divergence as well as gene expression polymorphism and divergence, protein-protein interactions cannot account for the observed coupling between regulatory and structural evolution. Furthermore, we show that proteins with higher rates of amino acid substitutions tend to have larger sizes and tend to be expressed at lower mRNA abundances, whereas genes with higher levels of gene expression divergence and polymorphism tend to have shorter sizes and tend to be expressed at higher mRNA abundances. Finally, we show that protein length is negatively associated with both number of protein-protein interactions and mRNA abundance and that interacting proteins in Drosophila show similar amounts of divergence. We suggest that protein sequences and gene expression are subjected to similar evolutionary dynamics, possibly because of similarity in the fitness effect (i.e., strength of stabilizing selection) of disruptions in a gene's protein sequence or its mRNA expression. We conclude that, as more and better data accumulate, understanding the causal connections among biological traits and how they are integrated over time to constrain or promote structural and regulatory evolution may finally become possible. PMID- 15746014 TI - More genes or more taxa? The relative contribution of gene number and taxon number to phylogenetic accuracy. AB - The relative contribution of taxon number and gene number to accuracy in phylogenetic inference is a major issue in phylogenetics and of central importance to the choice of experimental strategies for the successful reconstruction of a broad sketch of the tree of life. Maximization of the number of taxa sampled is the strategy favored by most phylogeneticists, although its necessity remains the subject of debate. Vast increases in gene number are now possible due to advances in genomics, but large numbers of genes will be available for only modest numbers of taxa, raising the question of whether such genome-scale phylogenies will be robust to the addition of taxa. To examine the relative benefit of increasing taxon number or gene number to phylogenetic accuracy, we have developed an assay that utilizes the symmetric difference tree distance as a measure of phylogenetic accuracy. We have applied this assay to a genome-scale data matrix containing 106 genes from 14 yeast species. Our results show that increasing taxon number correlates with a slight decrease in phylogenetic accuracy. In contrast, increasing gene number has a significant positive effect on phylogenetic accuracy. Analyses of an additional taxon-rich data matrix from the same yeast clade show that taxon number does not have a significant effect on phylogenetic accuracy. The positive effect of gene number and the lack of effect of taxon number on phylogenetic accuracy are also corroborated by analyses of two data matrices from mammals and angiosperm plants, respectively. We conclude that, for typical data sets, the number of genes utilized may be a more important determinant of phylogenetic accuracy than taxon number. PMID- 15746015 TI - Variation in the pattern of synonymous and nonsynonymous difference between two fungal genomes. AB - The proportion of synonymous nucleotide differences per synonymous site (p(S)) and the proportion of nonsynonymous differences per nonsynonymous site (p(N)) were computed at 1,993,217 individual codons in 4,133 protein-coding genes between the two yeast species Saccharomyces cerevisiae and Saccharomyces paradoxus. When the modified Nei-Gojobori method was used, significantly more codons with p(N) > p(S) were observed than expected, based on random pairing of observed p(S) and p(N) values. However, this finding was most likely explained by the presence of a strong negative correlation between the number of synonymous differences and the number of nonsynonymous differences at codons with at least one difference. As a result of this correlation, codons with p(N) > p(S) were characterized not only by unusually high p(N) but also by unusually low p(S). On the other hand, the number of codons with p(N)>p(S) (where p(S) is the mean p(S) for all codons) was very similar to the random expectation, and the observed number of 30-codon windows with p(N) > p(S) was significantly lower than the random expectation. These results imply that the occurrence of a certain number of codons or codon windows with p(N) > p(S) is expected given the nature of nucleotide substitution and need not imply the action of positive Darwinian selection. PMID- 15746016 TI - Selection on the structural stability of a ribosomal RNA expansion segment in Daphnia obtusa. AB - The high rate of sequence divergence in nuclear ribosomal RNA (rRNA) expansion segments offers a unique opportunity to study the importance of natural selection in their evolution. To this end, we polymerase chain reaction amplified and cloned a 589-nt fragment of the 18S rRNA gene containing expansion segments 43/e1 and 43/e4 from six individual Daphnia obtusa from four populations. We screened 2,588 clones using single-stranded conformation polymorphism analysis and identified 103 unique haplotype sequences. We detected two pairs of indel sites in segment 43/e4 that complement each other when the secondary structure of the linear sequence is formed. Seven of the 12 observed combinations of length variants at these four sites (haplotypes) are shared between individuals from different populations, which may suggest that some of the length variation was present in their common ancestor. Haplotypes with uncompensated indels were only observed at low frequencies, while compensated indel haplotypes were found at a wide range of frequencies, supporting the hypothesis that the energetic stability of expansion segments is a trait under natural selection. In addition, there was strong linkage disequilibrium between the four complementary indel sites, particularly those that pair with one another in the secondary structure. Despite selection against unpaired bulges at these four indel sites, some nucleotides that form unpaired bulges are highly conserved in segment 43/e4, indicating that they are under a different selective constraint, possibly due to their role in higher level structural interactions. PMID- 15746017 TI - Tertiary endosymbiosis driven genome evolution in dinoflagellate algae. AB - Dinoflagellates are important aquatic primary producers and cause "red tides." The most widespread plastid (photosynthetic organelle) in these algae contains the unique accessory pigment peridinin. This plastid putatively originated via a red algal secondary endosymbiosis and has some remarkable features, the most notable being a genome that is reduced to 1-3 gene minicircles with about 14 genes (out of an original 130-200) remaining in the organelle and a nuclear encoded proteobacterial Form II Rubisco. The "missing" plastid genes are relocated to the nucleus via a massive transfer unequaled in other photosynthetic eukaryotes. The fate of these characters is unknown in a number of dinoflagellates that have replaced the peridinin plastid through tertiary endosymbiosis. We addressed this issue in the fucoxanthin dinoflagellates (e.g., Karenia brevis) that contain a captured haptophyte plastid. Our multiprotein phylogenetic analyses provide robust support for the haptophyte plastid replacement and are consistent with a red algal origin of the chromalveolate plastid. We then generated an expressed sequence tag (EST) database of 5,138 unique genes from K. brevis and searched for nuclear genes of plastid function. The EST data indicate the loss of the ancestral peridinin plastid characters in K. brevis including the transferred plastid genes and Form II Rubisco. These results underline the remarkable ability of dinoflagellates to remodel their genomes through endosymbiosis and the considerable impact of this process on cell evolution. PMID- 15746018 TI - Age-related differences in multiple-object tracking. AB - Multiple-object tracking is the ability to attend (keep track of) the positions of multiple target items as they move among other items. The performance of young and older adults (M = 19 and 73 years old, respectively) was compared in two versions of a tracking task in which participants were required to monitor the positions of 1-4 moving targets in a field of 10 moving items. All participants were capable of tracking more than 1 item at once, but on average older participants tracked around 3 items at once whereas young adults tracked 4. Results suggest that there are age differences in the ability to either track or report positions of moving items. All participants reported positions of 1-4 targets in a field of 10 items with 100% accuracy when items were static. PMID- 15746019 TI - Neglect of dropout underestimates effects of death in longitudinal studies. AB - Investigations of terminal declines in mental abilities have assessed cognitive performance at a single time point and retrospectively compared survivors and decedents at a single later census date. Neglect of outcomes other than death, such as dropout, causes a loss of information on the relative frailty of survivors and deceased persons before the census date and on incidence of mortality and frailty among survivors after the census date. This discards information on differences in health status between younger and older survivors. The Heim AH4-1 intelligence test was given to 4,228 people between the ages of 42 and 92 years, and both deaths and dropouts were logged during three successive census periods during the subsequent 11 years. Within and across census periods, effects of impending death and dropout did not differ, decreasing with time from initial assessment. Thus the effects of terminal decline, or indeed of any other variable affecting cognitive performance, are miscalculated if dropout is ignored. PMID- 15746020 TI - Resilient or at risk? A 4-year study of older adults who initially showed high or low distress following conjugal loss. AB - Following conjugal loss, some people show relatively little distress for the first several months, whereas others show considerable distress. In this article we examine these patterns over a 4-year period. Drawing on prior research defining grief trajectories, we conducted repeated measures analyses of variance on data from 92 bereaved elders with one preloss and three postloss assessments. Findings demonstrated that those with low initial distress continued to do well up to 4 years postloss. Differential findings among those who showed high distress initially and over time suggested that this pattern remained chronic only for those who had reported high distress preloss. Results underscore the need to refine the criteria used to identify those who are at risk for long-term problems. PMID- 15746021 TI - Education and APOE-e4 in longitudinal cognitive decline: MacArthur Studies of Successful Aging. AB - Longitudinal data from the MacArthur Study of Successful Aging were used to test for interactions between education and apolipoprotein E (APOE) genotype with respect to time trends in cognitive performance. Interactions between education, APOE-e4 status, and time were found for overall cognitive function, and for subscales measuring memory and naming: The presence of the e4 allele was associated with steeper declines in cognition for those with a greater than eighth-grade education. For those with an eighth-grade education or less, time trends did not differ by APOE genotype. A measure of cognitive impairment (i.e., scores of < or = 7 on the Short Portable Mental Status Questionnaire) yielded parallel though weaker evidence for a similar interaction with respect to risk of cognitive impairment. These findings suggest that the presence of at least one e4 allele appears to reduce the protective effects of education for those with at least a ninth-grade education or more, resulting in steeper cognitive declines with age. PMID- 15746022 TI - Managing life through personal goals: intergoal facilitation and intensity of goal pursuit in younger and older adulthood. AB - Two studies varying in design (cross-sectional and longitudinal) and methods (questionnaires, diaries, and objective information) support the notion that personal goals are among the phenomena that show positive development throughout adulthood: Older adults (M = 64 years) reported more mutual facilitation among their personal goals and were more engaged in goal pursuit than were younger adults (M = 25 years). Results were robust when age-group differences in education and disposable time were controlled for, and they also emerged in a context where younger and older participants had one goal in common, namely, to start regular physical exercise. Mediational analyses showed that the older adults' higher intensity of goal pursuit was partly mediated by their higher level of intergoal facilitation. PMID- 15746023 TI - Intrusions in episodic recall: age differences in editing of overt responses. AB - Two experiments compared episodic word-list recall of young and older adults. In Experiment 1, using standard free-recall procedures, older adults recalled significantly fewer correct items and made significantly more intrusions (recall of items that had not appeared on the target list) than younger adults. In Experiment 2, we introduced a new method, called externalized free recall, in which participants were asked to recall any items that came to mind during the recall period but to indicate with an immediate key press those items they could identify as intrusions. Both age groups generated a large number of intrusions, but older adults were significantly less likely than young adults to identify these as nonlist items. Results suggest that an editing deficit may be a contributor to age differences in episodic recall and that externalized free recall may be a useful tool for testing computationally explicit models of episodic recall. PMID- 15746024 TI - Personality change precedes clinical diagnosis of dementia of the Alzheimer type. AB - We examined personality changes reported by a collateral source on the Blessed Dementia Scale in people who were nondemented when they entered a longitudinal study. Of the 108 participants examined here, 68 received a clinical diagnosis of dementia at some point after entry into the study. The other 40 participants died and came to autopsy with a clinical diagnosis of no dementia; however, 14 received a neuropathological diagnosis of Alzheimer's disease. The results indicate that initial personality changes often occur early, even earlier than clinical diagnosis. Individuals without a clinical diagnosis who showed presence of the disease at autopsy experienced personality changes comparable with those of individuals who had received a clinical diagnosis. Personality changes may aid in the early detection of dementia of the Alzheimer type, which could facilitate early treatment. PMID- 15746025 TI - Religious attendance and mortality: an 8-year follow-up of older Mexican Americans. AB - OBJECTIVE: Studies in the area of religion and mortality are based primarily on data derived from samples of predominantly non-Hispanic Whites. Given the importance of religion in the lives of Hispanics living in the United States, particularly older Hispanics, we examine the effects of religious attendance on mortality risk among Mexican Americans aged 65 and older. METHODS: We employ data from the Hispanic Established Populations for Epidemiologic Studies of the Elderly to predict the risk of all-cause mortality over an 8-year follow-up period. RESULT: Overall, the results show that those who attend church once per week exhibit a 32% reduction in the risk of mortality as compared with those who never attend religious services. Moreover, the benefits of weekly attendance persist with controls for sociodemographic characteristics, cardiovascular health, activities of daily living, cognitive functioning, physical mobility and functioning, social support, health behaviors, mental health, and subjective health. DISCUSSION: Our findings suggest that weekly church attendance may reduce the risk of mortality among older Mexican Americans. Future research should focus on identifying other potential mediators of the relationship between religious involvement and mortality risk in the Mexican-origin population. PMID- 15746026 TI - Effect of population aging on proportionate mortality from heart disease and cancer, U.S. 2000-2050. AB - OBJECTIVE: One-fourth of all deaths in the United States occur after age 85 (646,900 of 2,392,619 total deaths in 1999). By 2050 (assuming 1999 age-specific death rates), 47.4% of all deaths will occur after age 85 (2,994,935 of 6,314,725 total deaths). As we age, heart disease becomes increasingly important relative to cancer as a cause of death. Thus, as age at death is postponed, future populations will be dying of different age-specific causes. The study objective was to project the effect of population aging on future proportionate mortality rates for heart disease and cancer. METHODS: Using Census Bureau population projections and assuming 1999 age-specific death rates, the authors calculated the number of deaths from heart disease, cancer, and all causes in 10-year intervals from 2000 to 2050. RESULT: During this period, the total number of deaths for heart disease and cancer will increase 2.8-fold and 2.3-fold, respectively. The proportionate mortality rate for heart disease will increase from 30% to 33%, and the rate for cancer will drop from 23% to 20%. DISCUSSION: Changes in age distribution between now and 2050 will cause heart disease to increase its dominance over all other causes of death, and proportionate mortality for cancer will decline. PMID- 15746027 TI - Minor children and adult care exchanges with community-dwelling frail elders in a St. Lucian village. AB - OBJECTIVE: Research on care of community-dwelling frail elders typically includes formal health service providers and adult members of the informal care system. Involvement of children and adolescents with elder care is largely undocumented. The aim of this article is to describe children's involvement in elder care. These findings are part of an ethnographic community study that examined common Western assumptions about elder care in a St. Lucian village. METHODS: Data were obtained in a four-phase, 5-year, community-based ethnographic field study that included in-depth network analysis of elder households. RESULT: One hundred eighty-eight informal caregivers assisted 14 elder networks in obtaining the things they needed to live through provision of 355 care activities. Forty-five children (ages 3(1/2) to 16) provided 111 of 355 (31%) care activities. The frail elders gave adults and children community member caregivers 196 and 94 benefits, respectively. DISCUSSION: Minor children are integrally involved in reciprocal exchanges for elder care in this village. Although they do not provide all of the same care activities as adults, they clearly assist elders, especially with running errands. Elders emphasized different motivational mechanisms for involving minor children and adults in their care networks. PMID- 15746028 TI - Casino gambling among urban elders: just another social activity? AB - OBJECTIVES: This study explored older adults' attitudes about a range of activities and determined characteristics associated with casino participation. Activity theory was used as conceptual framework to examine casino gambling as a newer social activity and to identify characteristics related to gambling behavior among elders. METHODS: The sample consisted of 1,410 randomly selected participants, aged 60 and older, who reside in the city of Detroit. Through the use of a cross-sectional analyses, selected variables tested with casino visits included health, demographics, social activities, senior optimism, opinions about casinos, transportation, and social support network. RESULTS: Bivariate associations revealed that sociodemographic variables such as income, age, education, marriage, and transportation were significantly associated with casino participation. Additional factors associated with casino visitation included positive attitudes about casinos, social support network, and enjoying a variety of other "active" activities. DISCUSSION: The results of this study supported the theoretical perspective underlying the basis of this project. Although not a favorite activity, gambling is among several types of social activities in which older adults do participate. Results suggest further investigation of how casino activities relate to mental health and other leisure activities. PMID- 15746029 TI - African American grandparents raising grandchildren: a national study using the Census 2000 American Community Survey. AB - OBJECTIVE: The objective of this work was to determine the prevalence, sociodemographic characteristics, and service utilization patterns of African American grandparents raising grandchildren compared with noncaregiving peers. METHODS: Data were obtained from the Census 2000 Supplementary Survey/American Community Survey, a nationally representative survey of 890,000 households. Analysis was based upon comparison of 2,362 African American grandparent caregivers aged 45+ with 40,148 noncaregiving peers. Gender-specific analyses also were conducted. RESULT: Over 500,000 African Americans aged 45+ were estimated to be raising grandchildren in 2000. They were disproportionately female, younger, and less educated than noncaregivers and more likely to be living in poverty and receiving public assistance. Grandmother caregivers had significantly higher rates of functional limitations and poverty than either grandfather caregivers or other African American women aged 45+. DISCUSSION: African American grandparent caregivers, particularly grandmothers, represent a highly vulnerable population economically. The finding that four-fifths of African American grandmother caregivers below the poverty line were not receiving public assistance underscores the need for improving program outreach and substance to better serve this population. The use of theories of intersectionality and multiracial feminism in future studies of grandparent caregiving should enhance our understanding of how the impact of race on gender varies by social class. PMID- 15746031 TI - Nucleotide excision repair, oxidative damage, DNA sequence polymorphisms, and cancer treatment. PMID- 15746030 TI - The impact of childhood and adult SES on physical, mental, and cognitive well being in later life. AB - OBJECTIVES: To examine the relationships between socioeconomic status (SES) and health across the life course and their variations by gender and race/ethnicity. METHODS: The sample included 19,949 respondents aged 50 or over from the 1998 Health and Retirement Study. RESULTS: Lower childhood SES was associated with worse health outcomes in later life. Part of the effect of childhood SES on adult health occurred through childhood health. The impact of childhood SES on education and income in adulthood explained an even larger share of this effect. We also found a stronger effect of adult SES for those with lower childhood SES than for those with more advantaged childhoods. Moreover, childhood SES had a similar impact on health in later life for women and men and for Whites and non Whites. However, college education seemed more important for women's later health, whereas income seemed more important for men's health. Education appeared to have a weaker effect on adult health for Blacks and Hispanics than for Whites. DISCUSSION: Both childhood and adult SES are important for health. The negative impact of low childhood SES can be partially ameliorated if people from a low SES position during childhood mobilize to higher status in adulthood. PMID- 15746032 TI - Histologic evaluation of bronchial squamous lesions: any role in lung cancer risk assessment? PMID- 15746033 TI - Thrombopoietic factors in chronic bone marrow failure states: the platelet problem revisited. AB - Thrombocytopenia is a serious clinical problem in several different clinical settings. In chronic bone marrow failure states, which include aplastic anemia, myelodysplastic syndrome, and graft failure, the prolonged nature of thrombocytopenia often leads to alloimunization after repeated platelet transfusions, the consequence of which is a platelet-refractory state and enhanced risk of bleeding. Despite the introduction of several thrombopoietic factors into clinical trials, an effective way to alleviate thrombocytopenia has been elusive, and the problem in chronic bone marrow failure states has remained poorly addressed by clinical investigations. Even so, several studies by our group and others suggest that a subset of patients suffering from chronic bone marrow failure can respond to appropriate growth factor therapy. The temporal pace of response appears, however, to be much slower than that observed after administering growth factors which act on neutrophils. On the other hand, durable responses can be secured in some patients given thrombopoietic factors for long periods of time. Herein, we provide an overview of the clinical research investigations of thrombopoietic factors in chronic bone marrow failure, and the emerging insights these studies provide for understanding the process of thrombopoiesis and its therapy in this setting. PMID- 15746034 TI - Somatic mutations of epidermal growth factor receptor in colorectal carcinoma. AB - PURPOSE: Somatic mutations of the epidermal growth factor receptor (EGFR) gene may predict the sensitivity of non-small cell lung carcinoma to gefitinib. However, no mutations have been reported for colorectal carcinoma. We therefore analyzed EGFR mutations in colorectal adenocarcinomas by the combined use of laser microdissection and sequencing of genomic DNA. EXPERIMENTAL DESIGN: We examined 11 representative colorectal adenocarcinoma cell lines and 33 clinical samples of colorectal carcinoma. In the clinical cases, we carefully dissected only carcinoma cells from frozen sections by laser microdissection. After DNA extraction and PCR, we examined EGFR mutations by sequencing genomic DNA. RESULTS: None of 11 colorectal carcinoma cell lines exhibited somatic mutations, but 4 of 33 clinical tumors (12%) exhibited mutations in the EGFR kinase domain. This may be the first report of somatic mutations in colorectal adenocarcinoma. CONCLUSIONS: Our findings suggest that a distinct minority of colorectal adenocarcinomas exhibit somatic mutations of EGFR, and these tumors may be susceptible to gefitinib treatment. PMID- 15746035 TI - Characterization of CEBPA mutations in acute myeloid leukemia: most patients with CEBPA mutations have biallelic mutations and show a distinct immunophenotype of the leukemic cells. AB - PURPOSE: The transcription factor CCAAT/enhancer binding protein alpha, encoded by the CEBPA, is crucial for the differentiation of immature granulocytes. Mutation of the CEBPA may play an important role in leukemogenesis and prognosis. We sought to characterize the CEBPA mutation in acute myeloid leukemia (AML) and to clarify if there is a distinct immunophenotype for leukemic cells with the mutation. EXPERIMENT DESIGN: One hundred and four patients with de novo AML were evaluated for the CEBPA mutation and immunophenotype of the leukemic cells. RESULTS: Twenty-two distinct mutations were identified in 16 (15%) of 104 AML patients. Fourteen patients had biallelic mutations, mostly involving both the NH(2)-terminal TAD1 region and the COOH-terminal basic leucine zipper domain (bZIP). The mutations in the bZIP region were always tandem duplications and were located at hot-spot regions for topoisomerase II sites. Sequential study of the CEBPA mutations showed that the mutations disappeared at complete remission but the same mutations reappeared at relapse. None of the patients developed novel mutations during the follow-up period. Patients with CEBPA mutations had significantly higher incidences of CD7 (73%), CD15 (100%), CD34 (93%), and HLA-DR (93%) expression on the leukemic cells. CONCLUSION: These data revealed that most AML with CEBPA mutations were associated with an immunophenotype of HLA DR(+)CD7(+)CD13(+)CD14(-)CD15(+)CD33(+)CD34(+). The close relationship of CEBPA mutations with the leukemia status of the patients and the concordance of mutation in presenting and relapse samples implicate the CEBPA mutation as a potential marker for monitoring minimal residue disease. PMID- 15746036 TI - Down-regulation of LATS1 and LATS2 mRNA expression by promoter hypermethylation and its association with biologically aggressive phenotype in human breast cancers. AB - PURPOSE: LATS1 and LATS2 are tumor suppressor genes implicated in the regulation of cell cycle. Methylation status of the promoter regions of these genes as well as its correlation with their mRNA levels were studied in human breast cancers. Correlation of LATS1 and LATS2 mRNA levels with clinicopathologic characteristics of breast tumors were also studied. EXPERIMENTAL DESIGN: Methylation status of promoter regions of LATS1 and LATS2 was studied by a methylation-specific PCR and mRNA expression levels of LATS1 and LATS2 were determined by a real-time PCR assay in 30 breast cancers. In addition, correlation of LATS1 and LATS2 mRNA levels with clinicopathologic characteristics was studied in 117 breast cancers. RESULTS: Methylation-specific PCR showed that of 30 tumors, LATS1 promoter region was hypermethylated in 17 tumors (56.7%) and LATS2 promoter region was hypermethylated in 15 (50.0%) tumors. LATS1 mRNA levels in breast tumors with hypermethylation (2.15 +/- 0.37, mean +/- SE) were significantly (P < 0.01) lower than those without hypermethylation (6.09 +/- 1.38), and LATS2 mRNA levels in breast tumors with hypermethylation (1.42 +/- 0.66) were also significantly (P < 0.01) lower than those without hypermethylation (3.10 +/- 1.00). The decreased expression of LATS1 or LATS2 mRNA was significantly associated with a large tumor size, high lymph node metastasis, and estrogen receptor and progesterone receptor negativity. Furthermore, the decreased expression of LATS1 mRNA, but not LATS2 mRNA, was significantly (P < 0.05) associated with a poor prognosis. CONCLUSIONS: Hypermethylation of the promoter regions of LATS1 and LATS2 likely plays an important role in the down-regulation of their mRNA levels in breast cancers, and breast cancers with a decreased expression of LATS1 or LATS2 mRNA levels have a biologically aggressive phenotype. PMID- 15746037 TI - Expression of activated signal transducer and activator of transcription 3 predicts expression of vascular endothelial growth factor in and angiogenic phenotype of human gastric cancer. AB - PURPOSE: Angiogenic behavior is a critical aspect of cancer biology and subject to regulation by multiple molecular pathways. Because the signal transducer and activator of transcription 3 (Stat3) transcription factor regulates multiple genes important to angiogenesis, we sought to determine whether Stat3 expression is related to vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in gastric cancer and whether these factors predict survival in gastric cancer patients. EXPERIMENTAL DESIGN: The expression of Stat3 and VEGF was determined by immunohistochemistry using archival tissues from 86 cases of resected human gastric cancer and confirmed by Western blot analysis. Angiogenic phenotype was determined by CD34 staining and microvessel counting. RESULTS: Stat3 expression correlated with VEGF expression and MVD. In univariate survival analyses, Stat3 expression (P = 0.013) and MVD (P = 0.036) were associated with inferior survival. However, when Stat3 expression, VEGF expression, MVD, stage, completeness of resection, Lauren's histologic classification, and age were entered into a Cox proportional hazards model, only strong Stat3 expression (P = 0.049) and advanced stage (P < 0.01) were independently prognostic of poor survival. Furthermore, genetically enforced alterations of activated Stat3 expression led to altered VEGF expression and angiogenic potential in human gastric cancer cells. CONCLUSION: Dysregulated Stat3 activation may play an important role in VEGF overexpression and elevated angiogenic phenotype in gastric cancer and contribute to gastric cancer development and progression. PMID- 15746038 TI - Prospective study of quantitation of plasma DNA levels in the diagnosis of malignant versus benign prostate disease. AB - PURPOSE: The aim of this study was to determine the potential of cell-free DNA levels as a diagnostic marker for prostate cancer, having first established the effect that blood sample processing has on this measurement. EXPERIMENTAL DESIGN: A total of 152 blood samples were collected prospectively from patients before their prostate biopsy and 25 from men in two distinct control groups. Blood was processed to yield three components: one-spin plasma (n = 68), two-spin plasma (n = 152), and serum (n = 56) samples. RESULTS: Having established the effect of sample preparation on the measured DNA level, the more reliable two-spin plasma sample was used to determine the relationship between DNA and the presence of prostate cancer. Those patients with cancer (n = 78) had a significantly higher level of DNA compared with the control groups (P < 0.0001 and P < 0.0001). However, DNA levels in patients with a benign biopsy (n = 74) were significantly higher than the 78 patients confirmed to have cancer (P = 0.02). CONCLUSIONS: We conclude that the sample type used in the quantitation of cell-free DNA has an effect on the level reported. Elevated levels are present in the two-spin plasma samples of patients with prostate cancer compared with healthy controls but are not of diagnostic value during the management of prostate cancer. PMID- 15746039 TI - Alteration of gene expression in macroscopically normal colonic mucosa from individuals with a family history of sporadic colon cancer. AB - PURPOSE: We have shown that the expression of several genes associated with human colon cancer is altered in the morphologically normal colonic mucosa (MNCM) of APC(min) mice and humans with colon cancers. To determine whether these alterations also occur in the MNCM of individuals who have not developed colon cancer but are at high risk of doing so, we measured gene expression in the MNCM of individuals with a family history of colon cancer. METHODS: Expression of 16 genes in the MNCM of 12 individuals with a first-degree relative with sporadic colon cancer and 16 normal controls were measured by quantitative reverse transcription-PCR. All subjects tested had normal colonoscopic examinations. Biopsy samples of MNCM were obtained from the ascending, transverse, descending, and rectosigmoid regions of the colon (2-8 biopsy samples were obtained from each region). RESULTS: Relative to normal controls, the expression of several genes, including PPAR-gamma, SAA1, and IL-8 were significantly altered in the macroscopically normal rectosigmoid mucosa from individuals with a family history of colon cancer. CONCLUSIONS: Molecular abnormalities that precede the appearance of adenomatous polyp are present in the MNCM of individuals who have a family history of colon cancer. This observation raises the possibility of screening for individuals who are at an increased risk of developing colon cancer by analysis of gene expression in rectosigmoid biopsy samples. To assess this possibility, prospective studies will be needed to determine whether or not altered gene expression is associated with the subsequent development of adenomatous polyps and/ or colonic carcinomas. PMID- 15746040 TI - Nucleotide excision repair gene polymorphisms and recurrence after treatment for superficial bladder cancer. AB - PURPOSE: Interindividual differences in DNA repair capacity not only modify individual susceptibility to carcinogenesis, but also affect individual response to cancer treatment. Nucleotide excision repair (NER) is one of the major DNA repair pathways in mammalian cells involved in the removal of a wide variety of DNA lesions. Polymorphisms in NER genes may influence DNA repair capacity and affect clinical outcome of bladder cancer treatment. EXPERIMENTAL DESIGN: To test the influence of NER gene polymorphisms on superficial bladder cancer outcome (recurrence and progression), we conducted a follow-up study of 288 patients with superficial bladder cancer. Median follow-up among patients who were recurrence free at the end of observation was 21.7 months from diagnosis. The specific polymorphic loci examined include XPA [A/G at 5' untranslated region (UTR)], XPC (poly AT, Ala(499)Val, Lys(939)Gln), XPD (Asp(312)Asn, Lys(751)Gln), XPG (His(1104)Asp), ERCC 1 (G/T at 3' UTR), and ERCC6 (Met(1097)Val, Arg(1230)Pro). RESULTS: The ERCC6 (Met(1097)Val) polymorphism had a significant impact on recurrence: carriers of at least one variant allele (Val) had a significantly higher recurrence risk than carriers of the wild-type allele (Met/Met; hazard ratio, 1.54; 95% confidence interval, 1.02-2.33). There were no overall statistically significant differences in the distributions of the other polymorphisms between patients with and without recurrence. However, when we combined these variant genotypes, there was a significant trend for an increased recurrence risk with an increasing number of putative high-risk alleles. Using individuals with five or fewer putative high-risk alleles as the reference group, individuals with six to seven risk alleles and individuals with eight or more risk alleles had higher recurrence risks, with hazard ratios of 0.92 (0.54-1.57) and 2.53 (1.48-4.30), respectively (P for trend < 0.001). There was also a significant trend for shorter recurrence-free survival time with increasing number of variant alleles (log rank test, P = 0.0007). When we stratified the patients according to intravesical Bacillus Calmette-Guerin treatment, we found a significant trend for shorter recurrence-free survival time in patients with variant alleles of XPA or ERCC6 polymorphisms who received Bacillus Calmette Guerin treatment (log rank test, P = 0.078 and 0.022, respectively). There were no significant individual or joint associations between these polymorphisms and progression. CONCLUSIONS: These data suggest that interindividual differences in DNA repair capacity may have an important impact on superficial bladder cancer recurrence. A pathway-based approach is preferred to study the effects of individual polymorphism on clinical outcomes. PMID- 15746041 TI - Risk assessment in patients with acute myeloid leukemia and a normal karyotype. AB - PURPOSE: The recognition of a number of leukemia-specific cytogenetic abnormalities and their role as independent prognostic factors have provided considerable insights into leukemia pathogenesis and have paved the way to adopt risk-adapted treatment. However, approximately 50% of newly diagnosed acute myeloid leukemia (AML) have a normal karyotype. There has therefore been much interest in identifying molecular markers that could help to improve the prognostic stratification of patients with normal-karyotype AML. EXPERIMENTAL DESIGN: Consecutive untreated AML patients (n = 67) from a single institution all with normal karyotype were analyzed for the presence of mutations in the myeloid transcription factor gene CEBPA (for CCAAT/enhancer binding protein-alpha), for internal tandem duplications (ITD) of the tyrosine kinase receptor gene FLT3 (for fms-like tyrosine kinase 3), and for expression of the BAALC gene (for brain and acute leukemia, cytoplasmic). RESULTS: 17.9% of normal-karyotype AML had mutations in the CEBPA gene, and 28.4% had FLT3-ITD; 65.7% (44 of 67) had high BAALC expression and 34.3% (23 of 67) had low BAALC expression. Patients with CEBPA mutations had a very favorable course of their disease. Median disease-free survival (DFS) and overall survival (OS) were 33.5 and 45.5 months, respectively, compared with 10 (e.g., 12 months in patients without CEBPA mutations; P = 0.0017; P = 0.0007). AML patients with FLT3-ITD had significantly shorter median DFS (P = 0.0328) and OS (P = 0.0148) than patients without FLT3-ITD. High BAALC expression predicted for a shorter DFS (P = 0.0152) and OS (P = 0.0210) compared with AML with low BAALC expression; 53.7% of normal-karyotype AML had neither FLT3-ITD nor CEBPA mutations. We found that high BAALC expression in normal karyotype AML with neither FLT3-ITD nor CEBPA mutations (18 of 67) indicates adverse prognosis for both DFS and OS (P = 0.0001; e.g., P = 0.0001) compared with the group with low BAALC expression and absent FLT3-ITD and CEBPA mutations (18 of 67). Thus, BAALC expression represents a novel prognostic marker particularly for normal-karyotype AML patients with neither FLT3-ITD nor CEBPA mutations. CONCLUSIONS: Assessment of CEBPA mutations, FLT3-ITD, and BAALC expression permits to split normal-karyotype AML into clinically distinct subgroups. PMID- 15746042 TI - Pathologic assessment of melanoma sentinel nodes: a role for molecular analysis using quantitative real-time reverse transcription-PCR for MART-1 and tyrosinase messenger RNA. AB - PURPOSE: Molecular analysis of melanoma sentinel nodes (SN) is sensitive, but poorly specific because metastases cannot be distinguished from benign nevus inclusions (BNI). We investigated whether quantitative reverse transcription-PCR (RT-PCR) detection of MART-1 and tyrosinase mRNAs could improve this specificity and contribute to SN assessment. EXPERIMENTAL DESIGN: Two hundred twenty SNs from 95 melanoma patients analyzed by extensive immunohistopathology and real-time quantitative RT-PCR. RESULTS: Using histopathology, SNs and patients were allotted to three diagnostic groups: (a) metastasis positive, (b) BNI positive, and (c) melanocyte-free. Median MART-1 and tyrosinase mRNA levels in SNs were significantly different in patients with metastasis compared with patients with BNIs (P < 0.05) and patients without melanocytic lesions (P < 0.001). However, a "gray-zone" was observed where distinction, based on mRNA levels, could not be made between the three groups. For both genes, the highest mRNA level recorded in each RT-PCR-positive patient was positively correlated with Breslow's tumor thickness. For SNs with metastases, tumor burden was significantly correlated to the mRNA level. Using the presence of a MART-1 RT-PCR signal to detect patients with metastases, a sensitivity of 100% and a negative predictive value of 100% were achieved when extensive immunohistology was used as reference. CONCLUSIONS: Quantitative RT-PCR MART-1 and tyrosinase mRNA analysis cannot be used alone for SN diagnosis because of its poor specificity for melanoma metastasis. However, in approximately one third of cases without RT-PCR evidence of MART-1 expression, extensive histopathologic SN investigation is not necessary, thus substantially reducing the cost of SN analysis. The level of melanocyte-associated mRNA is associated with both tumor thickness and tumor burden as measured histopathologically, suggesting that this may be of prognostic value. PMID- 15746043 TI - Prognostic significance of vascular endothelial growth factor immunohistochemical expression in head and neck squamous cell carcinoma: a meta-analysis. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is considered as a prime mediator of angiogenesis. Various studies examining the relationship between VEGF protein overexpression with the clinical outcome in patients with head and neck squamous cell carcinoma have yielded inconclusive results. EXPERIMENTAL DESIGN: We conducted a meta-analysis of 12 studies (n = 1.002 patients) that evaluated the correlation between VEGF (detected by immunohistochemistry) and 2-year overall survival. The relation between VEGF and lymph node involvement (11 studies, n = 722) was also examined. Data were synthesized with random effect and fixed effect risk ratios. RESULTS: The estimated risk of death in 2 years was 1.88-fold higher in the VEGF-positive patients [95% confidence interval, 1.43 2.45; P < 0.001 random effect calculations]. Between-study heterogeneity was nonsignificant (P = 0.15) but larger studies tended to provide more conservative estimates (P = 0.097). VEGF overexpression was not significantly associated with the presence of lymph node metastasis (risk ratio, 1.20; 95% confidence interval, 0.97-1.49; P = 0.087) and there was significant between-study heterogeneity (P = 0.08). CONCLUSIONS: Although some modest bias cannot be excluded, VEGF positivity seems to be associated with worse overall survival in patients with head and neck squamous cell carcinoma. PMID- 15746044 TI - Serum mannan-binding lectin-associated serine protease 2 levels in colorectal cancer: relation to recurrence and mortality. AB - PURPOSE: Mannan-binding lectin-associated serine protease 2 (MASP-2) is a plasma protein involved in inflammatory processes. MASP-2 circulates in complex with the protein mannan-binding lectin (MBL) or ficolins, and is activated to recruit the complement system when MBL binds to its targets. The level of MASP-2 is genetically determined, and the aim of the present study was to evaluate the effect of MASP-2 levels on postoperative infection, recurrence and survival. EXPERIMENTAL DESIGN: MASP-2 concentrations were determined in serum from 605 patients collected before elective resection for primary colorectal cancer. The primary end points were postoperative infection, time to any recurrence, and time to death. The median time of follow-up was 7.9 years. RESULTS: MASP-2 levels were not correlated to postoperative infections (P = 0.49). High MASP-2 levels significantly correlated with recurrent cancer disease [P = 0.03; hazard ratio (HR) = 1.4; 95% confidence interval (CI), 1.0-2.0] and with poor survival (P = 0.0005; HR = 1.4; 95% CI, 1.2-1.7). Multivariate statistical analysis, including age, gender, Dukes' stage of disease, tumor localization, and postoperative pneumonia, showed that the MASP-2 level had an independent prognostic value in the patients (P = 0.0001; HR = 1.5; 95% CI, 1.2-1.8). CONCLUSION: In the cohort of patients with colorectal cancer investigated, MASP-2 concentration in serum proved to be an independent prognostic marker with high MASP-2 levels predicting recurrence and poor survival. Postoperative infection could not be shown to be associated with MASP-2 levels. PMID- 15746045 TI - Combined evaluation of expressions of cyclin E and p53 proteins as prognostic factors for patients with gastric cancer. AB - BACKGROUND: There is a lack of consistency regarding the prognostic value of cyclin E overexpression in gastric cancer (gastric cancer). Our aim was to report on this overexpression and to analyze its correlations with the clinicopathologic variables. Another aim was to examine if aberrant expression of both cyclin E and p53 might increase the malignant potential of gastric cancer. METHODS: Specimens from 89 patients with gastric cancer treated with "curative" intent were evaluated for cyclin E and p53 expressions using immunohistochemical method. The correlations between cyclin E overexpression alone or in combination with p53 expression and the patient's clinicopathologic variables were analyzed. RESULTS: Cyclin E overexpression and p53 expression were shown in 35 (39.3%) and 46 (51.7%) tumors, respectively. The incidence of cyclin E overexpression was significantly higher in deeply invasive cancers (P < 0.0001), in cancers with lymph node metastasis (P = 0.003), and in cancers with advanced stages (P < 0.0001). There were no significant correlations with other clinicopathologic variables. Patients in whom their tumors showed cyclin E overexpression alone or in combination with p53 survived less than patients with negative cyclin E tumors. Multivariate analysis revealed that combined cyclin E overexpression and p53 expression was significantly associated with poor survival after adjusting for other variables (hazard ratio, 3.12; P = 0.009). CONCLUSIONS: Cyclin E overexpression is a common event in gastric cancer. Gastric cancer with cyclin E overexpression exhibit increased aggressiveness in the presence of aberrant p53. The combination of cyclin E overexpression with the p53 expression in gastric cancer further distinguished a subgroup of patients with poor prognosis. PMID- 15746046 TI - An 19F magnetic resonance-based in vivo assay of solid tumor methotrexate resistance: proof of principle. AB - PURPOSE: Studies in oncology have implicated multiple molecular mechanisms as contributors to intrinsic and acquired tumor resistance to antifolate therapy. Here we show the utility of an (19)F-labeled methotrexate (FMTX) with (19)F magnetic resonance to differentiate between sensitive and resistant tumors in vivo and thus predict therapeutic response. EXPERIMENTAL DESIGN: Human sarcoma xenografts in nude mice were used in this study. The sarcoma cell lines chosen for this study (HT-1080, HS-16, and M-805) are well characterized in terms of their methotrexate sensitivity and molecular mechanisms of resistance. The pharmacokinetics of tumor uptake/washout of FMTX were monitored via in vivo (19)F magnetic resonance spectroscopy (pulse/acquire with surface coil localization) following an i.v. bolus injection. Response post-therapy, following leucovorin rescue, was monitored via tumor growth. RESULTS: The three tumor models show differences in both the peak concentrations of tumor FMTX and the dynamics of uptake/retention. These differences are most pronounced for time points late in the magnetic resonance observation period (225-279 minutes post-injection). A statistically significant linear correlation between tumor tissue concentrations of FMTX at these late time points and therapeutic response in the days/weeks post treatment is shown (R = 0.81, F = 9.27, P < 0.001). Interestingly, a 400 mg/kg i.v. bolus injection of FMTX is a more potent cytotoxic agent in vivo against methotrexate-sensitive tumors than is the parent compound (P = 0.011). CONCLUSIONS: In principle, the assay method described herein could be implemented in the clinic as a diagnostic tool to make decisions regarding therapeutic protocol for the treatment of osteosarcoma on a case-by-case basis. PMID- 15746047 TI - Prognostic effect of epidermal growth factor receptor and EGFRvIII in glioblastoma multiforme patients. AB - PURPOSE: The epidermal growth factor receptor (EGFR) is overexpressed in approximately 50% to 60% of glioblastoma multiforme tumors, and the most common EGFR mutant, EGFRvIII, is expressed in 24% to 67% of cases. We sought to determine whether glioblastoma multiforme expression of either overexpressed wild type EGFR or the mutant EGFRvIII is an independent predictor of overall patient survival. EXPERIMENTAL DESIGN: Glioblastoma multiforme patients (n = 196) underwent a > or =95% volumetric tumor resection followed by conformal radiation. Their EGFR and EGFRvIII status was determined by immunohistochemistry and survival analyses were done. RESULTS: In our study of glioblastoma multiforme patients, 46% (n = 91) failed to express EGFR, 54% (n = 105) had overexpression of the wild-type EGFR, and 31% (n = 61) also expressed the EGFRvIII. Patients within groups expressing the EGFR, EGFRvIII, or lacking EGFR expression did not differ in age, sex, Karnofsky performance scale score, extent of tumor resection, or radiation. The median overall survival times for patients with tumors having EGFR expression absent, overexpressed only, or mutant (EGFRvIII) were 0.96, 0.98, and 1.07 years, respectively. However, for patients surviving > or =1 year, these values were 2.03, 2.02, and 1.21 years (P < 0.0001; log-rank test comparing EGFRvIII with all others). This effect remained significant in the multivariate analysis after adjustment for all other cofactors including age and Karnofsky performance scale score (rate ratio 4.34; 95% confidence interval, 2.21-8.51). CONCLUSIONS: Neither the overexpressed wild-type EGFR nor EGFRvIII was an independent predictor of median overall survival in this selected cohort of patients who underwent extensive tumor resection. However, in patients surviving > or =1 year, the expression of EGFRvIII was an independent negative prognostic indicator. PMID- 15746048 TI - Outcome in Hodgkin's lymphoma can be predicted from the presence of accompanying cytotoxic and regulatory T cells. AB - PURPOSE: Recent studies of Hodgkin's lymphoma (HL) have suggested that the presence of regulatory T cells in the reactive background may explain the inhibition of the antitumoral host immune response observed in these patients. This study aimed to assess the relevance of regulatory T cells and CTLs present in the background of HL samples in the prognosis of a series of classic HL (cHL) patients. EXPERIMENTAL DESIGN: Expression of granzyme B and TIA-1 (markers for CTL) and FOXP3 (a marker for regulatory T cells) were evaluated independently by immunohistochemistry in tissue microarrays of 257 cHL patients and correlated with patient outcome. RESULTS: The combined influence of the presence of FOXP3(+) and TIA-1(+) cells distinguished three risk groups of patients with 5-year overall survival of 100%, 88%, and 73%. The presence of a small number of FOXP3(+) cells and a high proportion of TIA-1(+) cells in the infiltrate represent an independent prognostic factor that negatively influenced event-free survival and disease-free survival in cHL. Compared with the features at diagnosis, relapsed samples tended to have more TIA-1(+) cells and a lower proportion of FOXP3(+) cells in the reactive background. CONCLUSIONS: These data suggest that low infiltration of FOXP3(+) cells in conjunction with high infiltration of TIA-1(+) cells in cHL may represent biological markers predicting an unfavorable outcome. Moreover, the variation of these markers over the course of the disease implies a possible role for them in the progression of HL cases. PMID- 15746049 TI - A potent immunogenic general cancer vaccine that targets survivin, an inhibitor of apoptosis proteins. AB - We reported previously a HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL), recognized by CD8(+) CTL. This peptide was derived from survivin protein, an inhibitor of apoptosis proteins, expressed in a variety of tumors, such as adenocarcinoma, squamous cell carcinoma, and malignant melanoma. In this report, we provide further evidence that survivin-2B80-88 peptide might serve as a potent immunogenic cancer vaccine for various cancer patients. Overexpression of survivin was detected in surgically resected primary tumor specimens of most breast and colorectal cancers and some gastric cancers as assessed by immunohistochemical study. HLA-A24/survivin-2B80-88 tetramer analysis revealed that there existed an increased number of CTL precursors in peripheral blood mononuclear cells (PBMC) of HLA-A24(+) cancer patients, and in vitro stimulation of PBMCs from six breast cancer patients with survivin-2B80-88 peptide could lead to increases of the CTL precursor frequency. Furthermore, CTLs specific for this peptide were successfully induced from PBMCs in all 7 (100%) patients with breast cancers, 6 of 7 (83%) patients with colorectal cancers, and 4 of 7 (57%) patients with gastric cancers. These data indicate that survivin expressed in tumor tissues is antigenic in cancer patients, and survivin-2B80-88-specific CTLs are present in PBMCs of various cancer patients. Our study raises the possibility that this peptide may be applicable as a general cancer vaccine to a large proportion of HLA-A24(+) cancer patients. PMID- 15746050 TI - In vivo radioiodide imaging and treatment of breast cancer xenografts after MUC1 driven expression of the sodium iodide symporter. AB - PURPOSE: Expression of the sodium iodide symporter (NIS) in the thyroid gland provides for effective imaging and treatment of thyroid cancer using radiolabeled iodide. Transfer of NIS into other tumors would expand the utility of this treatment to tumors of nonthyroid origin. MUC1 is a transmembrane glycoprotein that is overexpressed in many tumor types, including breast, pancreatic, and ovarian. The aim of this study was to create a construct containing NIS under the control of the MUC1 promoter to target expression specifically to MUC1-positive breast cancer cells. EXPERIMENTAL DESIGN: A replication-deficient adenoviral construct was created containing the MUC1 promoter followed by the human NIS gene. Iodide uptake assays, Western blot, and immunohistochemistry were used to confirm NIS expression and function. Breast cancer xenografts in mice were infected with Ad5/MUC1/NIS and then imaged and treated using radioiodide. RESULTS: A 58-fold increase in iodide uptake was observed in infected MUC1 positive T47D cells with no significant increase observed in MUC1-negative MDA-MB 231 cells or in cells infected with the control virus. The in vivo study yielded clear images of Ad/MUC1/NIS-infected tumor xenografts using (123)I. Administration of a therapeutic dose of (131)I resulted in an 83% reduction in tumor volume, whereas control tumors continued to increase in size (P < 0.01). CONCLUSIONS: These results show that the MUC1 promoter is capable of directing efficient and selective expression of the NIS gene in MUC1-positive breast tumor cells. This could potentially have applications for both imaging and therapy in a range of MUC1-positive tumor types. PMID- 15746051 TI - B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides. AB - Human B cells detect CpG motifs within microbial DNA via TLR9. Synthetic CpG oligodeoxynucleotides are currently being tested in clinical trials for the therapy of different types of B cell non-Hodgkin's lymphoma. However, there is only limited information on the CpG oligodeoxynucleotide sensitivity of primary malignant B cells of different non-Hodgkin's lymphoma entities. Here we found that most B-cell malignancies except plasmacytoma respond to CpG oligodeoxynucleotides by up-regulating expression of costimulatory and antigen presenting molecules, by increasing expression of CD20, and by proliferation. In an in vitro analysis of 41 individual patient-derived primary tumor samples, B cell chronic lymphocytic leukemia (B-CLL) and marginal zone lymphoma showed the strongest activation upon stimulation with CpG oligodeoxynucleotides. Small lymphocytic lymphoma, follicular lymphoma, mantle cell lymphoma, and large cell lymphoma showed an intermediate response. Consistent with CpG oligodeoxynucleotides sensitivity, TLR9 mRNA was present in B-CLL but absent in plasmacytoma. Although CpG oligodeoxynucleotides induced proliferation in all CpG oligodeoxynucleotide-sensitive types of B-cell malignancies, proliferation was weaker than in normal B cells and at least for B-CLL was followed by increased apoptosis. In conclusion, B-cell malignancies show significant differences in their responsiveness to CpG oligodeoxynucleotides. Focusing clinical studies on patients with highly CpG oligodeoxynucleotide-sensitive B-cell malignancies may improve the clinical outcome of such trials. PMID- 15746052 TI - Endocrine effects of tamoxifen plus exemestane in postmenopausal women with breast cancer. AB - PURPOSE: In some specific circumstances, combined hormonal therapies for breast cancer seem to be more effective than single maneuvers. In two laboratory mammary cancer models, the combination of the aromatase inactivator exemestane plus tamoxifen gives a higher response rate than is found with either agent alone. To evaluate the endocrine effects of the combination of exemestane and tamoxifen, we studied 33 postmenopausal women disease-free following primary treatments for breast cancer who were taking tamoxifen for at least 3 months. DESIGN: After observation for symptoms on tamoxifen for 4 weeks, blood samples were taken and patients were begun additionally on exemestane 25 mg p.o. qd. Eight weeks later, blood samples were again taken, and exemestane was discontinued. RESULTS: A decrease in alkaline phosphatase was found with exemestane treatment (P = 0.06), whereas no change in osteocalcin level was observed. A decrease in high-density lipoprotein cholesterol level was found (P = 0.0025), whereas total cholesterol, low-density lipoprotein cholesterol and triglyceride levels showed no changes with exemestane treatment. Estradiol, estrone, and estrone sulfate levels decreased to immeasurable or very low levels with exemestane treatment (all P < 0.001). No significant changes in frequencies of common drug-associated side effects, such as vasomotor symptoms or weight change, were found. CONCLUSIONS: Based on the absence of adverse endocrine effects with the addition of exemestane to tamoxifen therapy observed in this study, further clinical evaluation of the efficacy of this combination is warranted. PMID- 15746053 TI - Phase I pharmacokinetic, food effect, and pharmacogenetic study of oral irinotecan given as semisolid matrix capsules in patients with solid tumors. AB - PURPOSE: To characterize the maximum-tolerated dose, recommended dose, dose limiting toxicities (DLT), pharmacokinetic profile, and food effect of orally administered irinotecan formulated as new semisolid matrix capsules. EXPERIMENTAL DESIGN: Irinotecan was given orally in fasted patients once daily for 5 consecutive days and repeated every 3 weeks. Patients were randomly assigned to take the drug along with a high-fat, high-calorie breakfast for the administration at day 1 of the first or second cycle. Dosages tested were 70 and 80 mg/m(2)/day. RESULTS: Twenty-five patients received 101 cycles of therapy (median two cycles, range 1-15). During the first cycle, grade 3 delayed diarrhea and grade 3 fever were the DLTs at the dosage of 80 mg/m(2)/day in three out of five patients. Hematologic and nonhematologic toxicities were mild to moderate. Exposure to the active metabolite SN-38 was relatively high compared with i.v. infusion, but no relevant accumulation was observed. Food had no significant effect on irinotecan pharmacokinetics. One confirmed partial remission and 10 disease stabilizations were observed in previously treated patients. No association was found between the UGT1A1*28 genotype and the risk of severe irinotecan-induced toxicity. CONCLUSIONS: For oral irinotecan, a dose of 70 mg/m(2)/day for 5 consecutive days every 3 weeks is recommended for further studies. Delayed diarrhea was the main DLT, similar to that observed with intravenously administered irinotecan. This study confirms that oral administration of irinotecan is feasible and may have favorable pharmacokinetic characteristics. PMID- 15746054 TI - Phase I study of MetXia-P450 gene therapy and oral cyclophosphamide for patients with advanced breast cancer or melanoma. AB - PURPOSE: MetXia-P450 is a novel recombinant retroviral vector that encodes the human cytochrome P450 type 2B6 gene (CYP2B6), Escherichia coli lacZ, and neomycin resistance marker genes. Cytochrome P450 enzymes are primarily expressed in the liver and convert the prodrug cyclophosphamide to an active phosphoramide mustard and acrolein. Gene-based delivery of CYP2B6 to the tumor site leads to local prodrug activation and higher concentrations of the active metabolites at the target site. EXPERIMENTAL DESIGN: MetXia-P450 was directly injected into metastatic cutaneous tumor nodules on days 1 and 2 and nodules biopsied on day 7. Oral cyclophosphamide (100 mg/m(2)) was administered between days 8 and 22. Subsequent cycles of oral cyclophosphamide were repeated for 2 of 4 weeks. Gene transfer levels in biopsy samples were measured by histologic and quantitative PCR analyses. Safety assessments were made using PCR for vector dissemination to the blood after injection and using PCR and serologic analyses to detect replicating virus. Secondary end points included clinical response, toxicity, and evaluation of antitumor immune responses by measurement of carcinoembryonic antigen and 5T4 antibodies. RESULTS: Twelve patients with breast cancer (n = 9) and melanoma (n = 3) received three dose levels of MetXia-P450 ( approximately 8 x 10(5), approximately 8 x 10(6), and approximately 8 x 10(7) lacZ transferring units/mL). The product was safe and well tolerated. The lacZ transgene was detected in biopsy material by immunohistochemistry in 10 of 12 patients and integrated viral sequences by PCR in 3 of 6 patients. One (8%) patient with breast cancer had a partial response and received 7 months of oral cyclophosphamide. Four (33%) patients had stable disease for > or =3 months and the rest had progressive disease. Preliminary immunologic analyses were suggestive of an antitumor response in two patients (partial response in one patient and stable disease in one patient). CONCLUSION: MetXia was safe and well tolerated. Gene transfer was detected at all dose levels, and the initial suggestion of an antitumor response indicates that MetXia-P450 should undergo further clinical assessment. PMID- 15746055 TI - T cell-dependent antibody responses against aberrantly expressed cyclin B1 protein in patients with cancer and premalignant disease. AB - PURPOSE: Cyclin B1-derived peptides were shown by us to be targets of tumor specific CD8(+) T cells in patients with breast and head and neck cancer. We obtained further evidence of cyclin B1 immunogenicity and its potential to serve as a tumor-specific antigen by analyzing its ability to elicit T cell-dependent humoral immune responses in vivo in patients with different types of tumors. EXPERIMENTAL DESIGN: Recombinant cyclin B1 protein from two different sources was purified and used as antigen in ELISA assays to test sera from patients with breast, pancreatic, colon, and lung cancer for the presence of anti-cyclin B1 antibody. We also analyzed patients with benign lung disease, premalignant disease, and a known history of heavy smoking. RESULTS AND CONCLUSIONS: Cyclin B1 elicits helper T cell-dependent antibody responses in vivo. Tumors with higher level of cyclin B1 expression elicit higher anti-cyclin B1 antibody levels. Antibodies in patients with breast and colon cancer are primarily of the IgG isotype whereas patients with pancreatic and lung cancer have in addition anti cyclin B1 IgA. Cyclin B1-specific IgG was also detected in long-term smokers and in patients with preneoplastic lung disease. Immune responses to aberrantly expressed cyclin B1 in tumors and premalignant lesions should be further explored as diagnostic and prognostic markers, in addition to their immunotherapeutic potential. PMID- 15746056 TI - Phase I trial of combretastatin a-4 phosphate with carboplatin. AB - PURPOSE: Preclinical evidence of synergy led to a phase I trial employing combretastatin A-4 phosphate (CA4P), a novel tubulin-binding antivascular drug, in combination with carboplatin. EXPERIMENTAL DESIGN: Based on preclinical scheduling studies, patients were treated on day 1 of a 21-day cycle. Carboplatin was given as a 30-minute i.v. infusion and CA4P was given 60 minutes later as a 10-minute infusion. RESULTS: Sixteen patients with solid tumors received 40 cycles of therapy at CA4P doses of 27 and 36 mg/m(2) together with carboplatin at area under the concentration-time curve (AUC) values of 4 and 5 mg min/mL. The dose-limiting toxicity of thrombocytopenia halted the dose escalation phase of the study. Four patients were treated at an amended dose level of CA4P of 36 mg/m(2) and carboplatin AUC of 4 mg min/mL although grade 3 neutropenia and thrombocytopenia were still observed. Three lines of evidence are adduced to suggest that a pharmacokinetic interaction between the drugs results in greater thrombocytopenia than anticipated: the carboplatin exposure (as AUC) was greater than predicted; the platelet nadirs were lower than predicted; and the deviation of the carboplatin exposure from predicted was proportional to the AUC of CA4, the active metabolite of CA4P. Patient benefit included six patients with stable disease lasting at least four cycles. CONCLUSION: This study of CA4P and carboplatin given in combination showed dose-limiting thrombocytopenia. Pharmacokinetic/pharmacodynamic modeling permitted the inference that altered carboplatin pharmacokinetics caused the increment in platelet toxicity. PMID- 15746057 TI - Polymorphisms in ERCC1 and grade 3 or 4 toxicity in non-small cell lung cancer patients. AB - PURPOSE: ERCC1 is a lead enzyme in the nucleotide excision repair pathway of DNA repair. Polymorphisms have been identified in the ERCC1 gene, the C8092A and codon 118 polymorphisms, which may lead to an altered capacity to regenerate damaged normal tissue and greater treatment-related toxicity. EXPERIMENTAL DESIGN: Using logistic regression models, we evaluated the ERCC1 C8092A and codon 118 polymorphisms and their association with the occurrence of grade 3 or 4 toxicity in 214 stage III and IV non-small cell lung cancer patients treated first line with platinum-based chemotherapy. Adjusting covariates were performance status and type of treatment regimen. RESULTS: There was no statistically significant association between either the C8092A or codon 118 polymorphism and overall or hematologic grade 3 or 4 toxicity. However, carrying at least one variant ERCC1 C8092A allele was associated with a significantly increased risk of grade 3 or 4 gastrointestinal toxicity (adjusted odds ratio, 2.33; 95% confidence interval, 1.07-5.05; P = 0.03). CONCLUSIONS: Adjusting for performance status and type of treatment regimen, carrying at least one ERCC1 8092A allele is associated with a >2-fold increase in grade 3 or 4 gastrointestinal toxicity among platinum-treated non-small cell lung cancer patients. PMID- 15746058 TI - Promoter hypermethylation of mismatch repair gene hMLH1 predicts the clinical response of malignant astrocytomas to nitrosourea. AB - PURPOSE: In certain types of human cancers, transcriptional inactivation of hMLH1 by promoter hypermethylation plays a causal role in the loss of mismatch repair functions that modulate cytotoxic pathways in response to DNA-damaging agents. The aim of the present study was to investigate the role of promoter methylation of the hMLH1 gene in malignant astrocytomas. EXPERIMENTAL DESIGN: We examined the hMLH1 promoter methylation in a homogeneous cohort of patients with 41 malignant astrocytomas treated by 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-2(2 chloroethyl)-3-nitrosourea chemotherapy in combination with radiation and interferon therapy, and assessed the correlation of such methylation with clinical outcome. RESULTS: hMLH1 promoter methylation was found in 6 (15%) of the 41 newly diagnosed malignant astrocytomas. Hypermethylation of the hMLH1 promoter corresponded closely with a loss of immunohistochemical staining for hMLH1 protein (P = 0.0013). Patients with hMLH1-methylated tumors displayed a greater chance of responding to adjuvant therapy as compared with those with hMLH1 unmethylated tumors (P = 0.0150). The presence of hMLH1 hypermethylation was significantly associated with a longer progression-free survival on both univariate analysis (P = 0.0340) and multivariate analysis (P = 0.0161). CONCLUSIONS: The present study identified hMLH1 methylation status as a predictor of the clinical response of malignant astrocytomas to chloroethylnitrosourea based adjuvant therapy. The findings obtained suggest that determination of the methylation status of hMLH1 could provide a potential basis for designing rational chemotherapeutic strategies, as well as for predicting prognosis. PMID- 15746059 TI - HA22 (R490A) is a recombinant immunotoxin with increased antitumor activity without an increase in animal toxicity. AB - PURPOSE: RFB4 (dsFv)-PE38 (BL22) is a recombinant immunotoxin containing an anti CD22 (Fv) fused to truncated Pseudomonas exotoxin A, which induces a high complete remission rate in patients with purine analogue-resistant hairy cell leukemia. HA22 is a mutant of BL22 with mutations in heavy-chain CDR3 resulting in increased cytotoxic activity. Our goal was to improve the activity of HA22. EXPERIMENTAL DESIGN: Arg(490), which is located in the catalytic domain (III) of the immunotoxin HA22, was mutated to alanine. Purified immunotoxins were produced and tested for cytotoxic activity in cell culture and for antitumor activity and nonspecific toxicity in mice. ADP-ribosylation activity was also measured. RESULTS: HA22 (R490A) is approximately 2-fold more cytotoxic than HA22 on several CD22-positive cell lines. When injected i.v., HA22 (R490A) has more potent antitumor activity than HA22 against CA46 tumors in mice. HA22 and HA22 (R490A) have similar LD(50)s (approximately 1.3 mg/kg) and similar plasma half-lives. The R490A mutation also improved the cytotoxicity of the antimesothelin recombinant immunotoxin SS1 (dsFv)-PE38 (SS1P). In vitro ADP-ribosylation assays show that HA22 R490A has increased activity. Increased cytotoxic activity is probably related to this increase in ADP-ribosylation activity. CONCLUSION: Protein engineering can be used to increase the efficacy of recombinant immunotoxins. Because HA22 (R490A) has increased antitumor activity without increased animal toxicity, immunotoxins with this mutation are candidates for clinical development. PMID- 15746060 TI - Antitumor effects of a monoclonal antibody that binds anionic phospholipids on the surface of tumor blood vessels in mice. AB - PURPOSE: We recently reported that anionic phospholipids, principally phosphatidylserine, become exposed on the external surface of viable vascular endothelial cells in tumors, possibly in response to oxidative stresses present in the tumor microenvironment. In the present study, we tested the hypothesis that a monoclonal antibody directed against anionic phospholipids might exert antitumor effects by causing vascular damage in tumors. EXPERIMENTAL DESIGN: A new mouse immunoglobulin G3 monoclonal antibody, 3G4, was raised that binds anionic phospholipids in the presence of serum or beta2-glycoprotein I. The antibody was tested for its ability to localize to tumor vessels and exert antitumor effects in mice. RESULTS: 3G4 recognized anionic phospholipids on the external membrane of H(2)O(2)-treated endothelial cells and in vitro. It localized specifically to tumor vascular endothelium and to necrotic tumor cells after injection into severe combined immunodeficient mice bearing orthotopic MDA MB-435 tumors. Treatment with 3G4 retarded the growth of four different tumors in mice. It reduced the growth of established orthotopic MDA-MB-231 and MDA-MB-435 human breast tumors in mice by 75% and 65% respectively, large L540 human Hodgkin's tumors by 50%, and small syngeneic Meth A fibrosarcomas by 90%. Histologic examination revealed vascular damage, a reduction in vascular density, and a reduction in tumor plasma volume. Treatment with 3G4 induced the binding of monocytes to tumor endothelium and infiltration of macrophages into MDA-MB-435 and MDA-MB-231 tumors. No toxicity to the mice was observed. CONCLUSIONS: 3G4 localizes specifically to complexes of anionic phospholipids and serum proteins on the surface of vascular endothelial cells in tumors in mice. This results in damage to tumor vasculature and suppression of tumor growth. PMID- 15746061 TI - The insulin-like growth factor-I receptor kinase inhibitor, NVP-ADW742, sensitizes small cell lung cancer cell lines to the effects of chemotherapy. AB - PURPOSE: Insulin-like growth factor-I (IGF-I) is a potent growth factor for small cell lung cancer (SCLC) in both the autocrine and endocrine context. It also inhibits chemotherapy-induced apoptosis through activation of the phosphatidylinositol 3-kinase (PI3K)-Akt pathway and we have previously shown that inhibition of this signaling pathway enhances sensitivity of SCLC cell lines to chemotherapy. The purpose of this study was to determine whether the novel IGF I receptor (IGF-IR) kinase inhibitor, NVP-ADW742, sensitizes SCLC cell lines to etoposide and carboplatin, which are commonly used in the treatment of SCLC. EXPERIMENTAL DESIGN: Cell growth in the presence of various combinations of NVP ADW742, imatinib (STI571; Gleevec/Glivec), and chemotherapeutic agents was monitored using a 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and analyzed using the Chou-Talalay multiple-drug-effect equation. Induction of apoptosis was assessed using terminal deoxynucleotide transferase mediated dUTP nick end labeling (TUNEL) and Western blot analysis of procaspase 3 and poly(ADP-ribose)polymerase cleavage. IGF-I-induced vascular endothelial cell growth factor expression was monitored by Northern blot and ELISA. RESULTS: NVP ADW742 synergistically enhanced sensitivity of multiple SCLC cell lines to etoposide and carboplatin. Maximal enhancement occurred at concentrations of NVP ADW742 that eliminated basal PI3K-Akt activity in individual cell lines. In the WBA cell line, in which the c-Kit receptor tyrosine kinase is partly responsible for basal PI3K-Akt activity, the combination of NVP-ADW742 and imatinib was superior to NVP-ADW742 alone in sensitizing the cells to etoposide. Enhancement of the sensitivity of SCLC cell lines to etoposide, as determined by MTT assay, correlated closely with sensitization to the induction of apoptosis as measured by TUNEL and caspase activation assays. Treatment with NVP-ADW742 also eliminated IGF-I-mediated expression of vascular endothelial cell growth factor, suggesting that in addition to enhancing sensitivity of SCLC to chemotherapy, this kinase inhibitor could potentially inhibit angiogenesis in vivo. CONCLUSIONS: Inhibition of IGF-IR signaling synergistically enhances the sensitivity of SCLC to etoposide and carboplatin. This enhancement in sensitivity to chemotherapy tightly correlates with inhibition of PI3K-Akt activation. Future SCLC clinical trials incorporating IGF-IR inhibitors alone or in combination with other kinase inhibitors should include assessment of PI3K-Akt activity as a pharmacodynamic end-point. PMID- 15746062 TI - Enhanced sensitivity to the HER1/epidermal growth factor receptor tyrosine kinase inhibitor erlotinib hydrochloride in chemotherapy-resistant tumor cell lines. AB - PURPOSE: Erlotinib (Tarceva, OSI-774) is a potent and specific inhibitor of the HER1/epidermal growth factor receptor (EGFR) tyrosine kinase. In phase II clinical studies, oral erlotinib monotherapy has shown antitumor activity in patients with advanced non-small cell lung cancer, head and neck cancer, and ovarian cancer after the failure of standard chemotherapy. We hypothesized that some tumors treated with multiple cytotoxic therapies may become more dependent on the HER1/EGFR signaling pathways for survival. EXPERIMENTAL DESIGN: The growth inhibitory effect of erlotinib was tested on 10 pairs of chemosensitive, parental, and chemoresistant tumor cell lines. RESULTS: Enhanced sensitivity to erlotinib was observed in the doxorubicin-resistant human breast cancer cell line MCF-7, paclitaxel-resistant human ovarian carcinoma cell line A2780, and cisplatin-resistant human cervical carcinoma cell line ME180. The IC(50) values of erlotinib in the resistant cell lines were 2- to 20-fold lower than those in the corresponding parental cell lines. This enhanced sensitivity to erlotinib correlated with higher HER1/EGFR and phospho-HER1/EGFR expression when compared with the corresponding parental cell lines. Acquired resistance to cytotoxic agents was not associated with cross-resistance to erlotinib. AE-ME180/CDDP resistant xenografts showed greater sensitivity to erlotinib than parental ME180 xenografts did. CONCLUSIONS: Our findings suggest that acquired resistance to cytotoxic therapy in some tumors is associated with enhanced sensitivity to HER1/EGFR inhibitors, which correlates with increased HER1/EGFR expression. These data may explain some of the observed clinical activity of HER1/EGFR inhibitors in patients previously treated with multiple therapies. HER1/EGFR tyrosine kinase inhibitors may be more effective as second- or third-line treatment for certain patients with tumors that were previously treated with multiple chemotherapy regimens. PMID- 15746063 TI - In vitro canine distemper virus infection of canine lymphoid cells: a prelude to oncolytic therapy for lymphoma. AB - PURPOSE: Measles virus (MV) causes the regression of human lymphoma xenografts. The purpose of this study was to determine if canine lymphoid cells could be infected in vitro with MV or canine distemper virus (CDV, the canine Morbillivirus equivalent of MV) and determine if in vitro viral infection leads to apoptotic cell death. EXPERIMENTAL DESIGN: Reverse transcriptase-PCR was used to examine the expression of both signal lymphocyte activation molecule (CD150) and membrane cofactor molecule (CD46) mRNA. An attenuated CDV expressing enhanced green fluorescent protein was used to infect canine cells in vitro. Both flow cytometry and reverse transcriptase-PCR was used to document CDV infection. Cell death was examined using a propidium iodide staining assay and Annexin V binding. RESULTS: Canine lymphoid cell lines and neoplastic B and T lymphocytes collected from dogs with spontaneous lymphoma expressed the Morbillivirus receptor CD150 mRNA. In contrast, only neoplastic lymphocytes expressed detectable levels of CD46 mRNA. Although MV did not infect canine cells, CDV efficiently infected between 40% and 70% of all three canine lymphoid lines tested. More importantly, CDV infected 50% to 90% of neoplastic lymphocytes isolated from dogs with both B and T cell lymphoma. Apoptosis of CDV-infected cell lines was documented. CONCLUSIONS: Attenuated CDV may be a useful treatment for canine lymphoma. As such, dogs with lymphoma may represent a biologically relevant large animal model to investigate the feasibility, safety, and efficacy of Morbillivirus therapy in a clinical setting with findings that may have direct applicability in the treatment of human non-Hodgkin's lymphoma. PMID- 15746064 TI - Patupilone acts as radiosensitizing agent in multidrug-resistant cancer cells in vitro and in vivo. AB - Interference with microtubule function is a promising antitumoral concept. Paclitaxel is a clinically validated tubulin-targeting agent; however, treatment with paclitaxel is often limited by taxane-related toxicities and is ineffective in tumors with multidrug-resistant cells. Patupilone (EPO906, epothilone B) is a novel non-taxane-related microtubule-stabilizing natural compound that retains full activity in multidrug-resistant tumors and is clinically less toxic than paclitaxel. Here we have investigated the effect of combined treatment with ionizing radiation and patupilone or paclitaxel in the P-glycoprotein overexpressing, p53-mutated human colon adenocarcinoma cell line SW480 and in murine, genetically defined E1A/ras-transformed paclitaxel-sensitive embryo fibroblasts. Patupilone and paclitaxel alone and in combination with ionizing radiation reduced the proliferative activity of the E1A/ras-transformed cell line with similar potency in the sub and low nanomolar range. SW480 cells were only sensitive to patupilone, and combined treatment with low-dose patupilone (0.1 nmol/L) followed by clinically relevant doses of ionizing radiation (2 and 5 Gy) resulted in a supra-additive cytotoxic effect. Inhibition of the drug efflux protein P-glycoprotein with verapamil resensitized SW480 cells to treatment with low doses of paclitaxel alone and in combination with IR. In tumor xenografts derived from SW480 cells a minimal treatment regimen with patupilone and fractionated irradiation (1 x 2 mg/kg plus 4 x 3 Gy) resulted in an at least additive tumor response with extended tumor growth arrest. Analysis by flow cytometry in vitro revealed an apoptosis- and G(2)-M-independent mode of radiosensitization by patupilone. Interestingly though, a transient accumulation of cells in S phase was observed on combined treatment.Overall, patupilone might be a promising alternative in paclitaxel-resistant, P-glycoprotein-overexpressing tumors for a combined treatment regimen using ionizing radiation and a microtubule inhibitor. PMID- 15746065 TI - Analyses of recombinant vaccinia and fowlpox vaccine vectors expressing transgenes for two human tumor antigens and three human costimulatory molecules. AB - PURPOSE: The poor immunogenicity of tumor antigens and the antigenic heterogeneity of tumors call for vaccine strategies to enhance T-cell responses to multiple antigens. Two antigens expressed noncoordinately on most human carcinomas are carcinoembryonic antigen (CEA) and MUC-1. We report here the construction and characterization of two viral vector vaccines to address these issues. EXPERIMENTAL DESIGN: The two viral vectors analyzed are the replication competent recombinant vaccinia virus (rV-) and the avipox vector, fowlpox (rF-), which is replication incompetent in mammalian cells. Each vector encodes the transgenes for three human costimulatory molecules (B7-1, ICAM-1, and LFA-3, designated TRICOM) and the CEA and MUC-1 transgenes (which also contain agonist epitopes). The vectors are designated rV-CEA/MUC/TRICOM and rF-CEA/MUC/TRICOM. RESULTS: Each of the vectors is shown to be capable of faithfully expressing all five transgenes in human dendritic cells (DC). DCs infected with either vector are shown to activate both CEA- and MUC-1-specific T-cell lines to the same level as DCs infected with CEA-TRICOM or MUC-1-TRICOM vectors. Thus, no evidence of antigenic competition between CEA and MUC-1 was observed. Human DCs infected with rV-CEA/MUC/TRICOM or rF-CEA/MUC/TRICOM are also shown to be capable of generating both MUC-1- and CEA-specific T-cell lines; these T-cell lines are in turn shown to be capable of lysing targets pulsed with MUC-1 or CEA peptides as well as human tumor cells endogenously expressing MUC-1 and/or CEA. CONCLUSION: These studies provide the rationale for the clinical evaluation of these multigene vectors in patients with a range of carcinomas expressing MUC-1 and/or CEA. PMID- 15746066 TI - Formation and antitumor activity of PNU-159682, a major metabolite of nemorubicin in human liver microsomes. AB - PURPOSE: Nemorubicin (3'-deamino-3'-[2''(S)-methoxy-4''-morpholinyl]doxorubicin; MMDX) is an investigational drug currently in phase II/III clinical testing in hepatocellular carcinoma. A bioactivation product of MMDX, 3'-deamino-3'',4' anhydro-[2''(S)-methoxy-3''(R)-oxy-4''-morpholinyl]doxorubicin (PNU-159682), has been recently identified in an incubate of the drug with NADPH-supplemented rat liver microsomes. The aims of this study were to obtain information about MMDX biotransformation to PNU-159682 in humans, and to explore the antitumor activity of PNU-159682. EXPERIMENTAL DESIGN: Human liver microsomes (HLM) and microsomes from genetically engineered cell lines expressing individual human cytochrome P450s (CYP) were used to study MMDX biotransformation. We also examined the cytotoxicity and antitumor activity of PNU-159682 using a panel of in vitro cultured human tumor cell lines and tumor-bearing mice, respectively. RESULTS: HLMs converted MMDX to a major metabolite, whose retention time in liquid chromatography and ion fragmentation in tandem mass spectrometry were identical to those of synthetic PNU-159682. In a bank of HLMs from 10 donors, rates of PNU 159682 formation correlated significantly with three distinct CYP3A-mediated activities. Troleandomycin and ketoconazole, both inhibitors of CYP3A, markedly reduced PNU-159682 formation by HLMs; the reaction was also concentration dependently inhibited by a monoclonal antibody to CYP3A4/5. Of the 10 cDNA expressed CYPs examined, only CYP3A4 formed PNU-159682. In addition, PNU-159682 was remarkably more cytotoxic than MMDX and doxorubicin in vitro, and was effective in the two in vivo tumor models tested, i.e., disseminated murine L1210 leukemia and MX-1 human mammary carcinoma xenografts. CONCLUSIONS: CYP3A4, the major CYP in human liver, converts MMDX to a more cytotoxic metabolite, PNU 159682, which retains antitumor activity in vivo. PMID- 15746067 TI - Effects of nonselective cyclooxygenase inhibition with low-dose ibuprofen on tumor growth, angiogenesis, metastasis, and survival in a mouse model of colorectal cancer. AB - PURPOSE: To determine whether the nonselective and relatively inexpensive nonsteroidal anti-inflammatory drug ibuprofen would be effective in inhibiting colorectal cancer and might improve mortality in a mouse model. EXPERIMENTAL DESIGN: The effects of ibuprofen on tumor growth inhibition and animal survival have been examined in both mouse and human colorectal cancer tumor models. Angiogenesis was measured by in vitro endothelial cell tube formation and immunohistochemistry. RESULTS: Ibuprofen significantly inhibited cell proliferation in mouse (MC-26) and human (HT-29) colorectal cancer cell lines. In vitro angiogenesis assays also indicated that ibuprofen decreased both cell proliferation and tube formation. The administration of chow containing 1,360 ppm ibuprofen, which achieved an average plasma concentration of ibuprofen lower than the peak level achieved in humans at therapeutic doses, inhibited tumor growth by 40% to 82%. Fewer liver metastases were found in the ibuprofen group compared with the control group. In combination therapy with the standard antineoplastic agents, 5-fluorouracil, or irinotecan (CPT-11), tumor volumes in the groups with ibuprofen +/- CPT-11 or 5-fluorouracil were smaller than in the control group. Ibuprofen was similar to the cyclooxygenase-2 selective inhibitor rofecoxib in its ability to suppress tumor growth and improve overall survival. CONCLUSIONS: Ibuprofen, in part by modulating tumor angiogenesis, decreases both tumor growth and metastatic potential in mice. The ibuprofen doses were in the low range of therapeutic human plasma concentrations. Ibuprofen potentiates the antitumor properties of CPT-11 and improves survival of mice without increasing gastrointestinal toxicity. PMID- 15746068 TI - Identification of CD19 and CD20 peptides for induction of antigen-specific CTLs against B-cell malignancies. AB - The purpose of these studies was to develop immunogenic peptides derived from the CD19 and CD20 self-antigens for the induction of antigen-specific CTLs against B cell malignancies. A total of seven peptides were designed and examined for their HLA-A2.1 affinity and immunogenicity. Of these peptides, we identified two highly immunogenic HLA-A2.1-specific peptides, CD19(150-158) (KLMSPKLYV) and CD20(188 196) (SLFLGILSV), which were capable of inducing peptide-specific CTLs. The CTLs displayed HLA-A2.1-restricted and antigen-specific cytotoxicity against Burkitt's lymphoma, chronic B cell leukemia, and multiple myeloma cell lines. The CD19 or CD20 peptide-specific CTL cytotoxicity was confirmed using HLA-A2.1(+) T2 cells presenting the appropriate peptide. No cytotoxic activity was observed against T2 cells presenting the irrelevant MAGE-3 peptide or T2 cells alone. In addition, the CTLs displayed a significant (P < 0.05) increase in cell proliferation and IFN-gamma secretion (>830 ng/mL) following restimulation with HLA A2.1(+)/CD19(+)/CD20(+) tumor cells. The CTLs also displayed a distinct phenotype consisting of a high percentage of CD69(+)/CD45RO(+) and a low percentage of CD45RA(+)/CCR7(+) CD4(+) or CD8(+) T cells characteristic of effector memory cell population. Cyclic guanosine 3',5'-monophosphate culture conditions using serum free AIM-V medium containing human AB serum, recombinant human interleukin 2 (Proleukin) and CD3/CD28 Dynabeads were developed resulting in a 35-fold expansion of CD20 peptide-specific CTLs. The expanded CD20-CTLs retained their cytotoxic activity (28-49%) against the Burkitt's lymphoma cell line. In conclusion, we report here on the identification of novel immunogenic CD19(150 158) (KLMSPKLYV) and CD20(188-196) (SLFLGILSV) peptides that have immunotherapeutic potentials as peptide vaccines or targeted T-cell therapies for treating B-cell malignancies. PMID- 15746069 TI - Induction of T-cell apoptosis in rats by genetically engineered glioma cells expressing granulocyte-macrophage colony-stimulating factor and B7.1. AB - PURPOSE: To evaluate antitumor effects on intracerebral gliomas of genetically engineered tumor vaccines expressing granulocyte-macrophage colony-timulating factor (GM-CSF), B7.1, or both (combination). EXPERIMENTAL DESIGN: A rat glioma cell line, RT-2, was engineered with a retroviral vector to express GM-CSF, B7.1, or combination. Tumorigenicity of engineered cells and therapeutic effects of s.c. given irradiated or live tumor vaccines on parental intracerebral gliomas were studied. Immune cell infiltration induced at vaccine and tumor sites was examined by histologic and immunohistochemical staining. Apoptosis of T cells from vaccine sites was analyzed with fluorescence-activated cell sorting. RESULTS: Engineered RT-2 cells exhibited reduced s.c. tumorigenicity in rats with reduced tumor growth and prolonged animal survival time compared with control rats. Rats with intracerebral gliomas s.c. treated with irradiated or live GM-CSF expressing vaccines had 60% and 100% survival rates, respectively, significantly better than the control groups (P < 0.05). In contrast, rats treated with vaccines expressing B7.1 or the combination had no or mild therapeutic effects. Studies revealed less T-cell infiltration at both vaccine and tumor sites in rats treated with vaccines expressing B7.1 or the combination than in rats treated with a vaccine expressing GM-CSF. Cell sorting analyses revealed higher proportions of apoptotic T cells at vaccine sites of rats treated with the combination than those treated with vaccine expressing GM-CSF. CONCLUSIONS: Combination of GM-CSF- and B7.1-expressing tumor vaccines exerted no synergistic, or even worse, therapeutic effects on gliomas compared with single GM-CSF secreting tumor vaccine. The worse therapeutic effects of the GM-B7.1-expressing tumor vaccine than the GM-CSF-expressing tumor vaccine were related to the reduced T-cell amount and increased T-cell apoptosis in the former. PMID- 15746070 TI - MEN4901/T-0128, a new camptothecin derivative-carboxymethyldextran conjugate, has potent antitumor activities in a panel of human tumor xenografts in nude mice. AB - PURPOSE: The purpose of the present study was to evaluate the antitumor activity and pharmacokinetic profile of MEN4901/T-0128 in nude mice bearing human tumor xenografts in comparison with irinotecan (CPT-11) and T-2513. EXPERIMENTAL DESIGN: We have determined the antitumor activity of MEN4901/T-0128, CPT-11, and T-2513 in BALB/cA Jcl nude mice bearing human gastric (H-81), colon (H-110), lung (Mqnu-1, H-74), esophageal (H-204), liver (H-181), and pancreatic (H-48) cancer lines, which had been serially transplanted s.c. and maintained in nude mice, and characterized the pharmacokinetic profile of MEN4901/T-0128 in nude mice bearing human gastric carcinoma St-4. RESULTS: MEN4901/T-0128 administered i.v. showed a marked antitumor activity in each of these tumor models, producing tumor shrinkage in the models of H-204 and H-181 carcinomas at its maximum tolerated dose of 80 mg/kg (expressed as T-2513) weekly for 4 weeks (q7d x 4) and tumor shrinking or marked growth-inhibitory effects in the models of H-81, H-110, Mqnu 1, H-74, and H-48 carcinomas at 1/3 of its maximum tolerated dose (q7d x 4). Pharmacokinetic analysis showed that MEN4901/T-0128 had an extended plasma half life with sustained tumor levels of T-2513, which may explain the superior activity of MEN4901/T-0128 in vivo. CONCLUSIONS: Because the efficacies of some drugs in this human cancer-nude mouse panel correlated well with their clinical outcomes in patients with the same type of cancers, the findings provide direct support that MEN4901/T-0128 is more efficacious than CPT-11 and is an excellent candidate for clinical trials for the treatment of solid tumors. PMID- 15746071 TI - Photochemical targeting of epidermal growth factor receptor: a mechanistic study. AB - PURPOSE: Photoimmunotherapy may allow target-specific photodynamic destruction of malignancies and may also potentiate anticancer antibody therapies. However, clinical use of either of the two modalities is limited for different reasons. Antibody therapies suffer from being primarily cytostatic and the need for prolonged administration with consequent side effects. In the case of photoimmunotherapy, a major impediment has been the absence of well-characterized photosensitizer immunoconjugates (PIC). In this investigation, we suggest a strategy to overcome these limitations and present the successful targeting of epidermal growth factor receptor (EGFR) using a well-characterized PIC. EXPERIMENTAL DESIGN: The PIC consisted of the EGFR-recognizing chimeric monoclonal antibody, C225, conjugated with a two-branched polyethylene glycol and benzoporphyrin derivative (BPD, Verteporfin). Mechanistic studies included photophysics, phototoxicity, cellular uptake, and catabolism experiments to yield dosimetric parameters. Target cells included two EGFR-overexpressing human cancer cell lines, OVCAR-5 and A-431. Nontarget cells included an EGFR-negative fibroblast cell line, 3T3-NR6, and a monocyte-macrophage cell line, J774. RESULTS: BPD-C225 PICs targeted and photodynamically killed EGFR-overexpressing cells, whereas free BPD exhibited no specificity. On a per mole basis, PICs were less phototoxic than free BPD, but PICs were very selective for target cells, whereas free BPD was not. Phototoxicity of the PICs increased at prolonged incubations. Photodynamic dose calculations indicated that PIC photophysics, photochemistry, catabolism, and subcellular localization were important determinants of PIC phototoxic potency. CONCLUSIONS: This study shows the efficacy of EGFR targeting with PIC constructs and suggests approaches to improve PIC designs and targeting strategies for in vivo photoimmunotherapy. The approach offers the possibility of dual effects via antibody-mediated cytostasis and photoimmunotherapy-based cytotoxicity. PMID- 15746072 TI - Plasma and cerebrospinal fluid pharmacokinetics of intravenous oxaliplatin, cisplatin, and carboplatin in nonhuman primates. AB - PURPOSE: Describe and compare the central nervous system pharmacology of the platinum analogues, cisplatin, carboplatin, and oxaliplatin and develop a pharmacokinetic model to distinguish the disposition of active drug from inert platinum species. EXPERIMENTAL DESIGN: Oxaliplatin (7 or 5 mg/kg), cisplatin (2 mg/kg), or carboplatin (10 mg/kg) was given i.v. Serial plasma and cerebrospinal fluid (CSF) samples were collected over 24 hours. Plasma ultrafiltrates were prepared immediately. Platinum concentrations were measured using atomic absorption spectrometry. Areas under the concentration x time curve were derived using the linear trapezoidal method. CSF penetration was defined as the CSF AUC(0 24)/plasma ultrafiltrate AUC(0-24) ratio. A four-compartment model with first order rate constants was fit to the data to distinguish active drug from inactive metabolites. RESULTS: The mean +/- SD AUCs in plasma ultrafiltrate for oxaliplatin, cisplatin, and carboplatin were 61 +/- 22, 18 +/- 6, and 211 +/- 64 micromol/L hour, respectively. The AUCs in CSF were 1.2 +/- 0.4 micromol/L hour for oxaliplatin, 0.56 +/- 0.08 micromol/L hour for cisplatin, and 8 +/- 2.2 mumol/L hour for carboplatin, and CSF penetration was 2.0%, 3.6%, and 3.8%, respectively. For oxaliplatin, cisplatin, and carboplatin, the pharmacokinetic model estimated that active drug accounted for 29%, 79%, and 81% of platinum in plasma ultrafiltrate, respectively, and 25%, 89%, and 56% of platinum in CSF, respectively. The CSF penetration of active drug was 1.6% for oxaliplatin, 3.7% for cisplatin, and 2.6% for carboplatin. CONCLUSIONS: The CSF penetration of the platinum analogues is limited. The pharmacokinetic model distinguished between active drug and their inactive (inert) metabolites in plasma and CSF. PMID- 15746073 TI - Green tea extract modulates actin remodeling via Rho activity in an in vitro multistep carcinogenic model. AB - Alteration of actin polymerization and loss of actin filaments is a marker of cellular dedifferentiation and early malignant transformation. To study this phenomenon, an in vitro human urothelial model consisting of two cell lines, HUC PC and MC-T11, were incorporated into the study design. These two cell lines have different malignant transformation potential. The effect of green tea extract (GTE), a potential anticancer agent, on actin remodeling was investigated. Upon exposure to the carcinogen 4-aminobiphenyl (4-ABP), the untransformed HUC-PC undergoes malignant transformation whereas the transformed MC-T11 progresses from noninvasive to invasive tumor. GTE induces actin polymerization in MC-T11 cells in a dose-responsive manner, but this effect is less obvious in the untransformed, more differentiated HUC-PC cells, which natively have higher actin polymerization status. In contrast, GTE antagonizes carcinogen 4-ABP induced actin depolymerization and stress fiber disruption in HUC-PC cells. In MC-T11 cells, GTE inhibits 4-ABP induced motility by increasing cell adhesion and focal adhesion complex formation. The effect of GTE on actin remodeling seems to be mediated by the stimulation of small GTP-binding protein Rho activity, because C3 exoenzyme, a specific inhibitor for Rho, blocks GTE-mediated Rho activation and stress fiber formation in MC-T11 cells. This study shows that GTE exerts an effect on cytoskeletal actin remodeling and provides further support for the use of GTE as a chemopreventive agent. PMID- 15746074 TI - The light reactions: a guide to recent acquisitions for the picture gallery. PMID- 15746075 TI - The open reading frame VI product of Cauliflower mosaic virus is a nucleocytoplasmic protein: its N terminus mediates its nuclear export and formation of electron-dense viroplasms. AB - The Cauliflower mosaic virus (CaMV) open reading frame VI product (P6) is essential for the viral infection cycle. It controls translation reinitiation of the viral polycistronic RNAs and forms cytoplasmic inclusion bodies (viroplasms) where virus replication and assembly occur. In this study, the mechanism involved in viroplasm formation was investigated by in vitro and in vivo experiments. Far protein gel blot assays using a collection of P6 deletion mutants demonstrated that the N-terminal alpha-helix of P6 mediates interaction between P6 molecules. Transient expression in tobacco (Nicotiana tabacum) BY-2 cells of full-length P6 and P6 mutants fused to enhanced green fluorescent protein revealed that viroplasms are formed at the periphery of the nucleus and that the N-terminal domain of P6 is an important determinant in this process. Finally, this study led to the unexpected finding that P6 is a nucleocytoplasmic shuttle protein and that its nuclear export is mediated by a Leu-rich sequence that is part of the alpha helix domain implicated in viroplasm formation. The discovery that P6 can localize to the nucleus opens new prospects for understanding yet unknown roles of this viral protein in the course of the CaMV infection cycle. PMID- 15746076 TI - High level of endothelial cell-specific gene expression by a combination of the 5' flanking region and the 5' half of the first intron of the VE-cadherin gene. AB - To develop a tool to obtain a high level of gene expression specifically in endothelial cells (ECs), we assessed enhancer activity of fragments in the first intron of the VE-cadherin gene using 3 different experimental systems: luciferase assay in the F2 EC line, green fluorescent protein (GFP) expression in ECs generated in embryonic stem (ES) cell differentiation culture, and GFP expression in transgenic mice. Although the 2.5-kbp (kilobase pair) 5' flanking sequence of the VE-cadherin gene is EC specific, adding 4 kbp of the 5' half of the first intron affected an enhancement of the gene expression level in all 3 assay systems. No other fragments tested in this study could confer such effects. Compared with other gene expression units, the unit described in this study would be the most optimum one available to date for EC-specific gene expression. Because this unit can express genes in VE-cadherin(+) progenitors of hematopoietic cells but not in fully committed hematopoietic cells, it will be useful to manipulate specifically the uncommitted progenitor stage during hematopoietic cell differentiation. PMID- 15746077 TI - Emergent autoimmunity in graft-versus-host disease. AB - Donor T-cell recognition of host alloantigens presented by host antigen presenting cells (APCs) is necessary for the induction of graft-versus-host disease (GVHD), but whether direct alloreactivity is sufficient for the propagation of GVHD is unknown. In this study, we demonstrate that GVHD cannot be effectively propagated through the direct pathway of allorecognition. Rather, donor T-cell recognition of antigens through the indirect pathway is necessary for the perpetuation of GVHD. Furthermore, GVHD results in the breaking of self tolerance, resulting in the emergence of donor T cells that can cause autoimmune disease in syngeneic recipients. Notably, GVHD-induced autoreactivity is donor APC dependent, transferable into secondary hosts, and involves cells of the innate immune system. These results indicate that donor T-cell--mediated pathologic damage during GVHD becomes donor APC dependent and provide a mechanistic explanation for the long-standing observation that GVHD is associated with autoimmune clinical manifestations. PMID- 15746078 TI - T-cell generation by lymph node resident progenitor cells. AB - In the thymus, 2 types of Lin-Sca-1+ (lineage-negative stem cell antigen-1 positive) progenitors can generate T-lineage cells: c-Kit(hi) interleukin-7 receptor alpha-negative (c-Kit(hi)IL-7Ralpha-) and c-Kit(lo)IL-7Ralpha+. While c Kit(hi)IL-7Ralpha- progenitors are absent, c-Kit(lo)IL-7Ralpha+ progenitors are abundant in the lymph nodes (LNs). c-Kit(lo)IL-7Ralpha+ progenitors undergo abortive T-cell commitment in the LNs and become arrested in the G1 phase of the cell cycle because they fail both to up-regulate c-myb, c-myc, and cyclin D2 and to repress junB, p16(INK4a), and p21(Cip1/WAF). As a result, development of LN c Kit(lo)IL-7Ralpha+ progenitors is blocked at an intermediate CD44+CD25lo development stage in vivo, and LN-derived progenitors fail to generate mature T cells when cultured with OP9-DL1 stromal cells. LN stroma can provide key signals for T-cell development including IL-7, Kit ligand, and Delta-like-1 but lacks Wnt4 and Wnt7b transcripts. LN c-Kit(lo)IL-7Ralpha+ progenitors are able to generate mature T cells when cultured with stromal cells producing wingless related MMTV integration site 4 (Wnt4) or upon in vivo exposure to oncostatin M whose signaling pathway intersects with Wnt. Thus, supplying Wnt signals to c Kit(lo)IL-7Ralpha+ progenitors may be sufficient to transform the LN into a primary T-lymphoid organ. These data provide unique insights into the essence of a primary T-lymphoid organ and into how a cryptic extrathymic T-cell development pathway can be amplified. PMID- 15746079 TI - In vitro and in vivo activity of ATP-based kinase inhibitors AP23464 and AP23848 against activation-loop mutants of Kit. AB - Oncogenic mutations of the Kit receptor tyrosine kinase occur in several types of malignancy. Juxtamembrane domain mutations are common in gastrointestinal stromal tumors, whereas mutations in the kinase activation loop, most commonly D816V, are seen in systemic mastocytosis and acute myelogenous leukemia. Kit activation-loop mutants are insensitive to imatinib mesylate and have been largely resistant to targeted inhibition. We determined the sensitivities of both Kit mutant classes to the adenosine triphosphate (ATP)-based inhibitors AP23464 and AP23848. In cell lines expressing activation-loop mutants, low-nM concentrations of AP23464 inhibited phosphorylation of Kit and its downstream targets Akt and signal transducer and activator of transcription 3 (STAT3). This was associated with cell-cycle arrest and apoptosis. Wild-type Kit-and juxtamembrane-mutant expressing cell lines required considerably higher concentrations for equivalent inhibition, suggesting a therapeutic window in which cells harboring D816V Kit could be eliminated without interfering with normal cellular function. Additionally, AP23464 did not disrupt normal hematopoietic progenitor-cell growth at concentrations that inhibited activation-loop mutants of Kit. In a murine model, AP23848 inhibited activation-loop mutant Kit phosphorylation and tumor growth. Thus, AP23464 and AP23848 potently and selectively target activation-loop mutants of Kit in vitro and in vivo and could have therapeutic potential against D816V-expressing malignancies. PMID- 15746080 TI - Telomere length as a prognostic parameter in chronic lymphocytic leukemia with special reference to VH gene mutation status. AB - B-cell chronic lymphocytic leukemia (CLL) consists of 2 prognostic entities where cases with mutated immunoglobulin V(H) genes have better outcome than unmutated cases. V(H)-mutated CLLs display longer telomeres compared with unmutated cases and telomere length has been indicated to predict outcome, although the prognostic value of telomere length has not been fully established in CLL. We analyzed telomere length, V(H) gene mutation status, and clinical parameters in a large series of CLL. Telomere length was assessed by quantitative polymerase chain reaction (PCR), giving a very good correlation to telomere length estimated by Southern blotting (P < .001). The prognostic information given by mutation status (n = 282) and telomere length (n = 246) was significant (P < .001, respectively). Telomere length was a prognostic factor for stage A (P = .021) and stage B/C (P = .018) patients, whereas mutation status predicted outcome only in stage A patients (P < .001). Furthermore, mutated CLLs were subdivided by telomere length into 2 groups with different prognoses (P = .003), a subdivision not seen for unmutated cases (P = .232). Interestingly, the V(H)-mutated group with short telomeres had an overall survival close to that of the unmutated cases. Thus, by combining V(H) mutation status and telomere length, an improved subclassification of CLL was achieved identifying previously unrecognized patient groups with different outcomes. PMID- 15746082 TI - Rapamycin selectively expands CD4+CD25+FoxP3+ regulatory T cells. AB - Rapamycin is an immunosuppressive compound that is currently used to prevent acute graft rejection in humans. In addition, rapamycin has been shown to allow operational tolerance in murine models. However, a direct effect of rapamycin on T regulatory (Tr) cells, which play a key role in induction and maintenance of peripheral tolerance, has not been demonstrated so far. Here, we provide new evidence that rapamycin selectively expands the murine naturally occurring CD4(+)CD25(+)FoxP3(+) Tr cells in vitro. These expanded Tr cells suppress proliferation of syngeneic T cells in vitro and prevent allograft rejection in vivo. Interestingly, rapamycin does not block activation-induced cell death and proliferation of CD4(+) T cells in vitro. Based on this new mode of action, rapamycin can be used to expand CD4(+)CD25(+)FoxP3(+) Tr cells for ex vivo cellular therapy in T-cell-mediated diseases. PMID- 15746081 TI - IFN-gamma-mediated negative feedback regulation of NKT-cell function by CD94/NKG2. AB - Activation of invariant natural killer T (iNKT) cells with CD1d-restricted T-cell receptor (TCR) ligands is a powerful means to modulate various immune responses. However, the iNKT-cell response is of limited duration and iNKT cells appear refractory to secondary stimulation. Here we show that the CD94/NKG2A inhibitory receptor plays a critical role in down-regulating iNKT-cell responses. Both TCR and NK-cell receptors expressed by iNKT cells were rapidly down-modulated by priming with alpha-galactosylceramide (alpha-GalCer) or its analog OCH [(2S,3S,4R)-1-O-(alpha-D-galactopyranosyl)-N-tetracosanoyl-2-amino-1,3,4 nonanetriol)]. TCR and CD28 were re-expressed more rapidly than the inhibitory NK cell receptors CD94/NKG2A and Ly49, temporally rendering the primed iNKT cells hyperreactive to ligand restimulation. Of interest, alpha-GalCer was inferior to OCH in priming iNKT cells for subsequent restimulation because alpha-GalCer induced interferon gamma (IFN-gamma) up-regulated Qa-1b expression and Qa-1b in turn inhibited iNKT-cell activity via its interaction with the inhibitory CD94/NKG2A receptor. Blockade of the CD94/NKG2-Qa-1b interaction markedly augmented recall and primary responses of iNKT cells. This is the first report to show the critical role for NK-cell receptors in controlling iNKT-cell responses and provides a novel strategy to augment the therapeutic effect of iNKT cells by priming with OCH or blocking of the CD94/NKG2A inhibitory pathway in clinical applications. PMID- 15746084 TI - Angiopoietin-1 promotes lymphatic sprouting and hyperplasia. AB - Angiopoietin 1 (Ang1), a ligand for the receptor tyrosine kinase Tie2, regulates the formation and stabilization of the blood vessel network during embryogenesis. In adults, Ang1 is associated with blood vessel stabilization and recruitment of perivascular cells, whereas Ang2 acts to counter these actions. Recent results from gene-targeted mice have shown that Ang2 is also essential for the proper patterning of lymphatic vessels and that Ang1 can be substituted for this function. In order to characterize the effects of the angiopoietins on lymphatic vessels, we employed viral vectors for overexpression of Ang1 in adult mouse tissues. We found that Ang1 activated lymphatic vessel endothelial proliferation, vessel enlargement, and generation of long endothelial cell filopodia that eventually fused, leading to new sprouts and vessel development. Cutaneous lymphatic hyperplasia was also detected in transgenic mice expressing Ang1 in the basal epidermal cells. Tie2 was expressed in the lymphatic endothelial cells and Ang1 stimulation of these cells resulted in up-regulation of vascular endothelial growth factor receptor 3 (VEGFR-3). Furthermore, a soluble form of VEGFR-3 inhibited the observed lymphatic sprouting. Our results reinforce the concept that Ang1 therapy may be useful in settings of tissue edema. PMID- 15746083 TI - Therapeutic potential of a tumor-specific, MHC-unrestricted T-cell receptor expressed on effector cells of the innate and the adaptive immune system through bone marrow transduction and immune reconstitution. AB - T-cell receptor (TCR) with unique major histocompatibility complex (MHC) unrestricted antigen-binding properties was isolated from a human T-cell clone specific for the tumor antigen MUC1. This TCR binds its epitope on the MUC1 protein without the requirement of processing and presentation. A single-chain Valpha/Vbeta/Cbeta (scTCR) was fused to a CD3 zeta (zeta) chain to allow expression on the surface of cells of the innate (granulocytes, macrophages, natural killer [NK] cells) as well as the adaptive (T and B cells) immune system. To test the ability of the cells of the innate immune system to reject a tumor when provided with a tumor antigen-specific TCR, we reconstituted severe combined immunodeficiency (SCID) mice with bone marrow cells transduced with a retroviral vector encoding this receptor and challenged them with a MUC1-positive human tumor. These mice controlled the growth of the tumor significantly better than the control mice. We performed a similar experiment in immunocompetent mice transgenic for human MUC1. Expression of the TCR on large percentages of cells did not result in infiltration or destruction of tissues expressing MUC1. Reconstituted mice controlled the outgrowth of a MUC1-transfected but not the parental control tumor. scTCR expression appears lifelong, suggesting a successful transduction of the self-renewing stem cells. PMID- 15746086 TI - Down syndrome in Down House: trisomy 21, GATA1 mutations, and Charles Darwin. PMID- 15746085 TI - Activation of Th1 and Tc1 cell adenosine A2A receptors directly inhibits IL-2 secretion in vitro and IL-2-driven expansion in vivo. AB - To evaluate the direct effect of adenosine on cytokine-polarized effector T cells, murine type 1 helper T cells (Th1) and type 1 cytotoxic T lymphocytes (Tc1) and Th2/Tc2 cells were generated using an antigen-presenting cell (APC) free method. Tc1 and Tc2 cells had similar adenosine signaling, as measured by intracellular cyclic AMP (cAMP) increase upon adenosine A(2A) receptor agonism by CGS21680 (CGS). CGS greatly reduced Tc1 and Tc2 cell interleukin 2 (IL-2) and tumor necrosis factor alpha (TNF-alpha) secretion, with nominal effect on interferon gamma (IFN-gamma) secretion. Tc2 cell IL-4 and IL-5 secretion was not reduced by CGS, and IL-10 secretion was moderately reduced. Agonist-mediated inhibition of IL-2 and TNF-alpha secretion occurred via A(2A) receptors, with no involvement of A(1), A(2B), or A(3) receptors. Adenosine agonist concentrations that abrogated cytokine secretion did not inhibit Tc1 or Tc2 cell cytolytic function. Adenosine modulated effector T cells in vivo, as CGS administration reduced CD4(+)Th1 and CD8(+)Tc1 cell expansion to alloantigen and, in a separate model, reduced antigen-specific CD4(+) Th1 cell numbers. Remarkably, agonist mediated T-cell inhibition was abrogated by in vivo IL-2 therapy. Adenosine receptor activation therefore preferentially inhibits type I cytokine secretion, most notably IL-2. Modulation of adenosine receptors may thus represent a suitable target primarily for inflammatory conditions mediated by Th1 and Tc1 cells. PMID- 15746087 TI - Lessened severe graft-versus-host after "minitransplantations". PMID- 15746088 TI - Clinical significance of intrahepatic hepatitis B virus covalently closed circular DNA in chronic hepatitis B patients who received cytotoxic chemotherapy. PMID- 15746089 TI - Dendritic cells have the option to express IDO-mediated suppression or not. PMID- 15746090 TI - Glycoprotein tertiary and quaternary structures are monitored by the same quality control mechanism. AB - Folding of glycoproteins entering the secretory pathway is strictly surveyed in the endoplasmic reticulum by a quality control system. Folding intermediates and proteins irreparably misfolded are marked via glucosylation by the UDPglucose:glycoprotein glucosyltransferase, an enzyme that acts as a folding sensor by exclusively labeling glycoproteins not displaying their native structures. Here we show that this sensing mechanism also applies to the oligomerization of protein complexes, as the glucosyltransferase appeared to be able to glucosylate folded complex subunits lacking the full complement of oligomer components. PMID- 15746091 TI - Transient receptor potential vanilloid type 1 activation down-regulates voltage gated calcium channels through calcium-dependent calcineurin in sensory neurons. AB - Calcium influx through voltage-activated Ca(2+) channels (VACCs) plays a critical role in neurotransmission. Capsaicin application inhibits VACCs and desensitizes nociceptors. In this study, we determined the signaling mechanisms of the inhibitory effect of capsaicin on VACCs in primary sensory neurons. Whole-cell voltage clamp recordings were performed in acutely isolated rat dorsal root ganglion neurons. Capsaicin caused a profound decrease in the Ca(2+) current (I(Ca)) density in capsaicin-sensitive, but not -insensitive, dorsal root ganglion neurons. At 1 mum, capsaicin suppressed about 60% of N-, P/Q-, L-, and R type I(Ca) density. Pretreatment with iodoresiniferatoxin, a specific transient receptor potential vanilloid type 1 (TRPV1) antagonist, or intracellular application of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid blocked the inhibitory effect of capsaicin on I(ca). However, neither W-7, a calmodulin blocker, nor KN-93, a CaMKII inhibitor, attenuated the inhibitory effect of capsaicin on I(Ca). Furthermore, intracellular dialysis of deltamethrin or cyclosporin A, the specific calcineurin (protein phosphatase 2B) inhibitors, but not okadaic acid (a selective protein phosphatase 1/protein phosphatase 2A inhibitor), abolished the effect of capsaicin on I(Ca). Interestingly, 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, deltamethrin, cyclosporin A, and okadaic acid each alone significantly increased the I(Ca) density and caused a depolarizing shift in the voltage dependence of activation. Immunofluorescence labeling revealed that capsaicin induced a rapid internalization of Ca(V)2.2 channels on the membrane. Thus, this study provides novel information that VACCs are tonically modulated by the intracellular Ca(2+) level and endogenous phosphatases in sensory neurons. Stimulation of TRPV1 by capsaicin down-regulates VACCs by dephosphorylation through Ca(2+)-dependent activation of calcineurin. PMID- 15746092 TI - p21Cip1 and p27Kip1 induce distinct cell cycle effects and differentiation programs in myeloid leukemia cells. AB - The cyclin-dependent kinase (Cdk) inhibitors p21(Cip1) and p27(Kip1) have been proposed to exert redundant functions in cell cycle progression and differentiation programs, although nonoverlapping functions have also been described. To gain further insights into the relevant mechanisms and to detect possible functional differences between both proteins, we conditionally expressed p21(Cip1) and p27(Kip1) in K562, a multipotent human leukemia cell line. Temporal ectopic expression of either p21(Cip1) or p27(Kip1) arrested proliferation, inhibited Cdk2 and Cdk4 activities, and suppressed retinoblastoma phosphorylation. However, whereas p21(Cip1) arrested cells in both G(1) and G(2) cell cycle phases, p27(Kip1) blocked the G(1)/S-phase transition. Furthermore, although both p21(Cip1) and p27(Kip1) associated with Cdk6, only p27(Kip1) significantly inhibited its activity. Most importantly, each protein promoted differentiation along a distinct pathway; p21(Cip1) triggered megakaryocytic maturation, whereas p27(Kip1) resulted in the expression of erythroid markers. Consistently, p21(Cip1) and p27(Kip1) were rapid and transiently up-regulated when K562 cells are differentiated into megakaryocytic and erythroid lineages, respectively. These findings demonstrate distinct functions of p21(Cip1) and p27(Kip1) in cell cycle regulation and differentiation and indicate that these two highly related proteins possess unique biological activities and are not functionally interchangeable. PMID- 15746093 TI - Constitutively active homo-oligomeric angiotensin II type 2 receptor induces cell signaling independent of receptor conformation and ligand stimulation. AB - Members of the G-protein-coupled receptor superfamily (GPCRs) undergo homo- and/or hetero-oligomerization to induce cell signaling. Although some of these show constitutive activation, it is not clear how such GPCRs undergo homo oligomerization with transmembrane helix movement. We previously reported that angiotensin II (Ang II) type 2 (AT(2)) receptor, a GPCR, showed constitutive activation and induced apoptosis independent of its ligand, Ang II. In the present study, we analyzed the translocation and oligomerization of the AT(2) receptor with transmembrane movement when the receptor induces cell signaling. Constitutively active homo-oligomerization, which was due to disulfide bonding between Cys(35) in one AT(2) receptor and Cys(290) in another AT(2) receptor, was localized in the cell membrane without Ang II stimulation and induced apoptosis without changes in receptor conformation. These results provide the direct evidence that the constitutively active homo-oligomeric GPCRs by intermolecular interaction in two extracellular loops is translocated to the cell membrane and induces cell signaling independent of receptor conformation and ligand stimulation. PMID- 15746094 TI - Interference with phosphoenzyme isomerization and inhibition of the sarco endoplasmic reticulum Ca2+ ATPase by 1,3-dibromo-2,4,6-tris(methylisothiouronium) benzene. AB - ATP hydrolysis and Ca(2+) transport by the sarco-endoplasmic reticulum Ca(2+) ATPase (SERCA) are inhibited by 1,3-dibromo-2,4,6-tris(methylisothiouronium) benzene (Br(2)-TITU) in the micromolar range (Berman, M. C., and Karlish, S. J. (2003) Biochemistry 42, 3556-3566). In a study of the mechanism of inhibition, we found that Br(2)-TITU allows the enzyme to bind Ca(2+) and undergo phosphorylation by ATP. The level of ADP-sensitive phosphoenzyme (i.e. E1P 2Ca(2+)) observed in the transient state following addition of ATP is much higher in the presence than in the absence of the inhibitor. Br(2)-TITU does not interfere with enzyme phosphorylation by P(i) in the reverse direction of the cycle (i.e. E2P) and produces only a slight inhibition of its hydrolytic cleavage. The inhibitory effect of Br(2)-TITU on steady state ATPase velocity is attributed to interference with the E1P-2Ca(2+) to E2P-2Ca(2+) transition. In fact, experiments on conformation-dependent protection from proteolytic digestion suggest that, in the presence of Br(2)-TITU, the loops connecting the "A" domain to the ATPase transmembrane region undergo greater fluctuation than expected in the E2 and E2P states. Optimal stability of the gathered headpiece domains is thereby prevented. These effects are opposite to those of thapsigargin, in which the mechanism of inhibition is related to stabilization of a highly compact ATPase conformation and interference with Ca(2+) binding and phosphoenzyme formation. Our experiments with Br(2)-TITU provide the first demonstration of a kinetic limit posed by an inhibitor on the E1P-2Ca(2+) to E2P-2Ca(2+) transition in the wild-type enzyme. PMID- 15746095 TI - Selective uncoupling of G alpha 12 from Rho-mediated signaling. AB - The heterotrimeric G protein G(12) has been implicated in such cellular regulatory processes as cytoskeletal rearrangement, cell-cell adhesion, and oncogenic transformation. Although the activated alpha-subunit of G(12) has been shown to interact directly with a number of protein effectors, the roles of many of these protein-protein interactions in G(12)-mediated cell physiology are poorly understood. To begin dissecting the specific cellular pathways engaged upon G(12) activation, we produced a series of substitution mutants in the regions of Galpha(12) predicted to play a role in effector binding. Here we report the identification and characterization of an altered form of Galpha(12) that is functionally uncoupled from signaling through the monomeric G protein Rho, a protein known to propagate several Galpha(12)-mediated signals. This mutant of Galpha(12) fails to bind the Rho-specific guanine nucleotide exchange factors p115RhoGEF and LARG (leukemia-associated RhoGEF), fails to stimulate Rho dependent transcriptional activation, and fails to trigger activation of RhoA and the Rho-mediated cellular responses of cell rounding and c-jun N-terminal kinase activation. Importantly, this mutant of Galpha(12) retains coupling to the effector protein E-cadherin, as evidenced by its ability both to bind E-cadherin in vitro and to disrupt E-cadherin-mediated cell-cell adhesion. Furthermore, this mutant retains the ability to trigger beta-catenin release from the cytoplasmic domain of cadherin. This identification of a variant of Galpha(12) that is selectively uncoupled from one signaling pathway while retaining signaling capacity through a separate pathway will facilitate investigations into the mechanisms through which G(12) proteins mediate diverse biological responses. PMID- 15746096 TI - Structural rationale for the affinity of pico- and femtomolar transition state analogues of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase. AB - Immucillin and DADMe-Immucillin inhibitors are tight binding transition state mimics of purine nucleoside phosphorylases (PNP). 5'-Methylthioadenosine/S adenosylhomocysteine nucleosidase (MTAN) is proposed to form a similar transition state structure as PNP. The companion paper describes modifications of the Immucillin and DADMe-Immucillin inhibitors to better match transition state features of MTAN and have led to 5'-thio aromatic substitutions that extend the inhibition constants to the femtomolar range (Singh, V., Evans, G. B., Lenz, D. H., Mason, J., Clinch, K., Mee, S., Painter, G. F., Tyler, P. C., Furneaux, R. H., Lee, J. E., Howell, P. L., and Schramm, V. L. (2005) J. Biol. Chem. 280, 18265-18273). 5'-Methylthio-Immucillin A (MT-ImmA) and 5'-methylthio-DADMe Immucillin A (MT-DADMe-ImmA) exhibit slow-onset inhibition with K(i)(*) of 77 and 2 pm, respectively, and were selected for structural analysis as the parent compounds of each class of transition state analogue. The crystal structures of Escherichia coli MTAN complexed with MT-ImmA and MT-DADMe-ImmA were determined to 2.2 A resolution and compared with the existing MTAN inhibitor complexes. These MTAN-transition state complexes are among the tightest binding enzyme-ligand complexes ever described and analysis of their mode of binding provides extraordinary insight into the structural basis for their affinity. The MTAN-MT ImmA complex reveals the presence of a new ion pair between the 4'-iminoribitol atom and the nucleophilic water (WAT3) that captures key features of the transition state. Similarly, in the MTAN-MT-DADMe-ImmA complex a favorable hydrogen bond or ion pair interaction between the cationic 1'-pyrrolidine atom and WAT3 is crucial for tight affinity. Distance analysis of the nucleophile and leaving group show that MT-ImmA is a mimic of an early transition state, while MT DADMe-ImmA is a better mimic of the highly dissociated transition state of E. coli MTAN. PMID- 15746097 TI - Presenilin-dependent gamma-secretase processing regulates multiple ERBB4/HER4 activities. AB - Transmembrane receptors typically transmit cellular signals following growth factor stimulation by coupling to and activating downstream signaling cascades. Reports of proteolytic processing of cell surface receptors to release an intracellular domain (ICD) has raised the possibility of novel signaling mechanisms directly mediated by the receptor ICD. The receptor tyrosine kinase ERBB4/HER4 (referred to here as ERBB4) undergoes sequential processing by tumor necrosis factor-alpha converting enzyme and presenilin-dependent gamma-secretase to release the ERBB4 ICD (4ICD). Our recent data suggests that regulation of gene expression by the ERBB4 nuclear protein and the proapoptotic activity of ERBB4 involves the gamma-secretase release of 4ICD. To determine the role gamma secretase processing plays in ERBB4 signaling, we generated an ERBB4 allele with the transmembrane residue substitution V673I (ERBB4-V673I). We demonstrate that ERBB4-V673I fails to undergo processing by gamma-secretase but retains normal cell surface signaling activity. In contrast to wild-type ERBB4, however, ERBB4 V673I was excluded from the nuclei of transfected cells and failed to activate STAT5A stimulation of the beta-casein promoter. These results support the contention that gamma-secretase processing of ERBB4 is necessary to release a functional 4ICD nuclear protein which directly regulates gene expression. We also demonstrate that 4ICD failed to accumulate within mitochondria of ERBB4-V673I transfected cells and the potent proapoptotic activity of ERBB4 was completely abolished in cells expressing ERBB4-V673I. Our results provide the first formal demonstration that proteolytic processing of ERBB4 is a critical event regulating multiple receptor signaling activities. PMID- 15746098 TI - NF90 regulates cell cycle exit and terminal myogenic differentiation by direct binding to the 3'-untranslated region of MyoD and p21WAF1/CIP1 mRNAs. AB - NF90 and splice variant NF110/ILF3/NFAR are double-stranded RNA-binding proteins that regulate gene expression. Mice with targeted disruption of NF90 were engineered. NF90(-/-) mice were born small and weak and succumbed to perinatal death within 12 h because of neuromuscular respiratory failure. Lung inflation and morphology were normal in NF90(-/-) mice. The diaphragm and other skeletal muscles in NF90(-/-) mice demonstrated disorganized arrangement and paucity of myofibers, evidence of myocyte degeneration and increased apoptosis. The expression of myogenic regulators, MyoD, myogenin, and p21WAF1/CIP1, was severely decreased in NF90(-/-) mice. These myogenic transcription factors and cell cycle inhibitors are regulated in part through post-transcriptional mRNA stabilization. Northwestern blotting revealed that NF90 is the principal and specific p21WAF1/CIP1 and MyoD 3'-untranslated region RNA-binding protein in developing skeletal muscles. NF90 regulates transcription factors and a cell cycle inhibitor essential for skeletal muscle differentiation and for survival. PMID- 15746099 TI - Chronic airway infection/inflammation induces a Ca2+i-dependent hyperinflammatory response in human cystic fibrosis airway epithelia. AB - Hyperinflammatory responses to infection have been postulated as a component of cystic fibrosis (CF) lung disease. Studies have linked intracellular calcium (Ca(2+)(i)) mobilization with inflammatory responses in several systems. We have reported that the pro-inflammatory mediator bradykinin (BK) promotes larger Ca(2+)(i) signals in CF compared with normal bronchial epithelia, a response that reflects endoplasmic reticulum (ER)/Ca(2+) store expansion induced by chronic luminal airway infection/inflammation. The present study investigated whether CF airway epithelia were hyperinflammatory and, if so, whether the hyperinflammatory CF phenotype was linked to larger Ca(2+) stores in the ER. We found that DeltaF508 CF bronchial epithelia were hyperinflammatory as defined by an increased basal and mucosal BK-induced interleukin (IL)-8 secretion. However, the CF hyperinflammation expressed in short-term (6-11-day-old) primary cultures of DeltaF508 bronchial epithelia was lost in long-term (30-40-day-old) primary cultures of DeltaF508 bronchial epithelia, indicating this response was independent of mutant cystic fibrosis transmembrane conductance regulator. Exposure of 30-40-day-old cultures of normal airway epithelia to supernatant from mucopurulent material (SMM) from CF airways reproduced the increased basal and mucosal BK-stimulated IL-8 secretion of short-term CF cultures. The BK-triggered increased IL-8 secretion in SMM-treated cultures was mediated by an increased Ca(2+)(i) mobilization consequent to an ER expansion associated with increases in protein synthesis (total, cytokines, and antimicrobial factors). The increased ER dependent, Ca(2+)(i)-mediated hyperinflammatory epithelial response may represent a general beneficial airway epithelial adaptation to transient luminal infection. However, in CF airways, the Ca(2+)(i)-mediated hyperinflammation may be ineffective in promoting the eradication of infection in thickened mucus and, consequently, may have adverse effects in the lung. PMID- 15746100 TI - Progressive adipocyte hypertrophy in aquaporin-7-deficient mice: adipocyte glycerol permeability as a novel regulator of fat accumulation. AB - Aquaporin-7 (AQP7) is a water/glycerol transporting protein expressed in adipocyte plasma membranes. We report here remarkable age-dependent hypertrophy in adipocytes in AQP7-deficient mice. Wild type and AQP7 null mice had similar growth at 0-16 weeks as assessed by body weight; however, by 16 weeks AQP7 null mice had 3.7-fold increased body fat mass. Adipocytes from AQP7 null mice of age 16 weeks were greatly enlarged (diameter 118 mum) compared with wild type mice (39 mum). Adipocytes from AQP7 null mice also accumulated excess glycerol (251 versus 86 nmol/mg of protein) and triglycerides (3.4 versus 1.7 mumol/mg of protein). In contrast, at age 4 weeks, adipocyte volume and body fat mass were comparable in wild type and AQP7 null mice. To investigate the mechanism(s) responsible for the progressive adipocyte hypertrophy, glycerol permeability and fat metabolism were studied in adipocytes isolated from the younger mice. Plasma membrane glycerol permeability measured by [(14)C]glycerol uptake was 3-fold reduced in AQP7-deficient adipocytes. However, adipocyte lipolysis, measured by free fatty acid release and hormone-sensitive lipase activity, and lipogenesis, measured by [(14)C]glucose incorporation into triglycerides, were not affected by AQP7 deletion. These data suggest that adipocyte hypertrophy in AQP7 deficiency results from defective glycerol exit and consequent accumulation of glycerol and triglycerides. Increasing AQP7 expression/function in adipocytes may reduce adipocyte volume and fat mass in obesity. PMID- 15746101 TI - Crystal structures of the RNA-dependent RNA polymerase genotype 2a of hepatitis C virus reveal two conformations and suggest mechanisms of inhibition by non nucleoside inhibitors. AB - Crystal structures of the RNA-dependent RNA polymerase genotype 2a of hepatitis C virus (HCV) from two crystal forms have been determined. Similar to the three dimensional structures of HCV polymerase genotype 1b and other known polymerases, the structures of the HCV polymerase genotype 2a in both crystal forms can be depicted in the classical right-hand arrangement with fingers, palm, and thumb domains. The main structural differences between the molecules in the two crystal forms lie at the interface of the fingers and thumb domains. The relative orientation of the thumb domain with respect to the fingers and palm domains and the beta-flap region is altered. Structural analysis reveals that the NS5B polymerase in crystal form I adopts a "closed" conformation that is believed to be the active form, whereas NS5B in crystal form II adopts an "open" conformation and is thus in the inactive form. In addition, we have determined the structures of two NS5B polymerase/non-nucleoside inhibitor complexes. Both inhibitors bind at a common binding site, which is nearly 35 A away from the polymerase active site and is located in the thumb domain. The binding pocket is predominantly hydrophobic in nature, and the enzyme inhibitor complexes are stabilized by hydrogen bonding and van der Waals interactions. Inhibitors can only be soaked in crystal form I and not in form II; examination of the enzyme-inhibitor complex reveals that the enzyme has undergone a dramatic conformational change from the form I (active) complex to the form II (inactive). PMID- 15746102 TI - Structural clues to Rab GTPase functional diversity. AB - Rab GTPases are key regulators of membrane trafficking in eukaryotes. Recent structural analysis of a number of Rabs, either alone or in complex with partner proteins, has provided new insight into the importance of both conserved and non conserved features of these proteins that specify their unique functions and localizations. This review will highlight what we have learned from crystallographic analysis of this important protein family. PMID- 15746103 TI - Molecular dissection of the interaction between p27 and Kip1 ubiquitylation promoting complex, the ubiquitin ligase that regulates proteolysis of p27 in G1 phase. AB - The cyclin-dependent kinase (CDK) inhibitor p27 is degraded at the G(0)-G(1) transition of the cell cycle by the ubiquitin-proteasome pathway in a Skp2 independent manner. We recently identified a novel ubiquitin ligase, KPC (Kip1 ubiquitylation-promoting complex), consisting of KPC1 and KPC2, which regulates the ubiquitin-dependent degradation of p27 at G(1) phase. We have now investigated the structural requirements for the interactions of KPC1 with KPC2 and p27. The NH(2)-terminal region of KPC1 was found to be responsible for binding to KPC2 and to p27. KPC1 mutants that lack this region failed to mediate polyubiquitylation of p27 in vitro and expression of one such mutant delayed p27 degradation in vivo. We also generated a series of deletion mutants of p27 and found that KPC failed to polyubiquitylate a p27 mutant that lacks the CDK inhibitory domain. Interestingly, the cyclin E.CDK2 complex prevented both the interaction of KPC with p27 as well as KPC-mediated polyubiquitylation of p27. A complex of cyclin E with a kinase-negative mutant of CDK2 also exhibited these inhibitory effects, suggesting that cyclin E.CDK2 competes with KPC1 for access to the CDK inhibitory domain of p27. These results suggest that free p27 is recognized by the NH(2)-terminal region of KPC1, which also associates with KPC2, and that p27 is then polyubiquitylated by the COOH-terminal RING-finger domain of KPC1. PMID- 15746104 TI - Mg2+ and Ca2+ differentially regulate DNA binding and dimerization of DREAM. AB - DREAM (calsenilin/KChIP3) is an EF-hand calcium-binding protein that represses transcription of prodynorphin and c-fos genes. Here we present structural and binding studies on single-site mutants of DREAM designed to disable Ca(2+) binding to each of the functional EF-hands (EF-2: D150N; EF-3: E186Q; and EF-4: E234Q). Isothermal titration calorimetry (ITC) analysis of Ca(2+) binding to the various mutants revealed that, in the absence of Mg(2+), Ca(2+) binds independently and sequentially to EF-3 (DeltaH = -2.4 kcal/mol), EF-4 (DeltaH = +5.2 kcal/mol), and EF-2 (DeltaH = +1 kcal/mol). By contrast, only two Ca(2+) bind to DREAM in the presence of physiological levels of Mg(2+) for both wild type and D150N, suggesting that EF-2 binds constitutively to Mg(2+). ITC measurements demonstrate that one Mg(2+) binds enthalpically with high affinity (K(d) = 13 mum and DeltaH = -0.79 kcal/mol) and two or more Mg(2+) bind entropically in the millimolar range. Size-exclusion chromatography studies revealed that Mg(2+) stabilizes DREAM as a monomer, whereas Ca(2+) induces protein dimerization. Electrophoretic mobility shift assays indicated that Mg(2+) is essential for sequence-specific binding of DREAM to DNA response elements (DREs) in prodynorphin and c-fos genes. The EF-hand mutants bind specifically to DRE, suggesting they are functionally intact. None of the EF-hand mutants bind DRE at saturating Ca(2+) levels, suggesting that binding of a single Ca(2+) at either EF-3 or EF-4 is sufficient to drive conformational changes that abolish DNA binding. NMR structural analysis indicates that metal-free DREAM adopts a folded yet flexible molten globule-like structure. Both Ca(2+) and Mg(2+) induce distinct conformational changes, which stabilize tertiary structure of DREAM. We propose that Mg(2+) binding at EF-2 may structurally bridge DREAM to DNA targets and that Ca(2+)-induced protein dimerization disrupts DNA binding. PMID- 15746105 TI - Exosite-interactive regions in the A1 and A2 domains of factor VIII facilitate thrombin-catalyzed cleavage of heavy chain. AB - Thrombin catalyzes the proteolytic activation of factor VIII, cleaving two sites in the heavy chain and one site in the light chain of the procofactor. Evaluation of thrombin binding the reaction products from heavy chain cleavage by steady state fluorescence energy transfer using a fluorophore-labeled, active site modified thrombin as well as by solid phase binding assays using a thrombin Ser(205) --> Ala mutant indicated a high affinity site in the A1 subunit (K(d) approximately 5 nm) that was dependent upon the Na(+)-bound form of thrombin, whereas a moderate affinity site in the A2 subunit (K(d) approximately 100 nm) was observed for both Na(+)-bound and -free forms. The solid phase assay also indicated that hirudin blocked thrombin interaction with the A1 subunit and had little, if any, effect on its interaction with the A2 subunit. Conversely, heparin blocked thrombin interaction with the A2 subunit and showed a marginal effect on A1 binding. Evaluation of the A2 sequence revealed two regions rich in acidic residues that are localized close to the N and C termini of this domain. Peptides encompassing these clustered acidic regions, residues 373-395 and 719 740, blocked thrombin cleavage of the isolated heavy chain at Arg(372) and Arg(740) and inhibited A2 binding to thrombin Ser(205) --> Ala, suggesting that both A2 domain regions potentially support interaction with thrombin. A B domainless, factor VIII double mutant Asp(392) --> Ala/Asp(394) --> Ala was constructed, expressed, and purified and possessed specific activity equivalent to a severe hemophilia phenotype. This mutant was resistant to cleavage at Arg(740), whereas cleavage at Arg(372) was not affected. These data suggest the acidic region comprising residues 389-394 in factor VIII A2 domain interacts with thrombin via its heparin-binding exosite and facilitates cleavage at Arg(740) during procofactor activation. PMID- 15746107 TI - Mortality in adult cardiac surgery. PMID- 15746106 TI - Incidence and risk factors for non-alcoholic steatohepatitis: prospective study of 5408 women enrolled in Italian tamoxifen chemoprevention trial. AB - OBJECTIVE: To assess the incidence, cofactors, and excess risk of development of non-alcoholic fatty liver disease, including non-alcoholic steatohepatitis, attributable to tamoxifen in women. DESIGN: Prospective, randomised, double blind, placebo controlled trial. SETTING AND PARTICIPANTS: 5408 healthy women who had had hysterectomies, recruited into the Italian tamoxifen chemoprevention trial from 58 centres in Italy. INTERVENTION: Women were randomly assigned to receive tamoxifen (20 mg daily) or placebo for five years. MAIN OUTCOME MEASURE: Development of non-alcoholic fatty liver disease in all women with normal baseline liver function who showed at least two elevations of alanine aminotransferase (> or = 1.5 times upper limit of normal) over a six month period. RESULTS: During follow up, 64 women met the predefined criteria: 12 tested positive for hepatitis C virus, and the remaining 52 were suspected of having developed non-alcoholic fatty liver disease (34 tamoxifen, 18 placebo)- hazard ratio = 2.0 (95% confidence interval 1.1 to 3.5; P = 0.04). In all 52 women ultrasonography confirmed the presence of fatty liver. Other factors associated with the development of non-alcoholic fatty liver disease included overweight (2.4, 1.2 to 4.8), obesity (3.6, 1.7 to 7.6), hypercholesterolaemia (3.4, 1.4 to 7.8), and arterial hypertension (2.0, 1.0 to 3.8). Twenty women had liver biopsies: 15 were diagnosed as having mild to moderate steatohepatitis (12 tamoxifen, 3 placebo), and five had fatty liver alone (1 tamoxifen, 4 placebo). No clinical, biochemical, ultrasonic, or histological signs suggestive of progression to cirrhosis were observed after a median follow up of 8.7 years. CONCLUSIONS: Tamoxifen was associated with higher risk of development of non alcoholic steatohepatitis only in overweight and obese women with features of metabolic syndrome, but the disease, in both the tamoxifen and the placebo group, after 10 years of follow up seems to be indolent. PMID- 15746108 TI - Data protection and freedom of information. PMID- 15746109 TI - Bat rabies in the United Kingdom. PMID- 15746110 TI - HIV testing. PMID- 15746111 TI - Simulation based training. PMID- 15746112 TI - NICE proposes to withdraw Alzheimer's drugs from NHS. PMID- 15746113 TI - UK stocks up on antiviral drug to tackle flu outbreak. PMID- 15746114 TI - UN committee approves declaration on human cloning. PMID- 15746115 TI - Abstinence programmes do not reduce HIV prevalence in Uganda. PMID- 15746116 TI - Mother found guilty in case of fabricated illness. PMID- 15746117 TI - Pathologist stands by his findings in Sally Clark case. PMID- 15746119 TI - A fifth of elderly people in Israel are abused. PMID- 15746122 TI - Poor information hampers discussion of HIV "super strain". PMID- 15746123 TI - Bush signs act that restricts class action suits in state courts. PMID- 15746125 TI - Two class actions against tobacco industry get the go-ahead. PMID- 15746128 TI - Breast cancer detection rates increase but coverage varies. PMID- 15746129 TI - More than 40% of cancer patients not seen by specialist in two weeks. PMID- 15746131 TI - Mortality data in adult cardiac surgery for named surgeons: retrospective examination of prospectively collected data on coronary artery surgery and aortic valve replacement. AB - OBJECTIVES: To present named surgeon mortality for isolated first time coronary artery surgery and aortic valve surgery. DESIGN: Retrospective analysis of prospectively collected data. SETTING: All NHS hospitals undertaking adult cardiac surgery in north west England. PARTICIPANTS: 25 consultant surgeons carrying out coronary artery surgery and aortic valve replacement between April 2001 and March 2004. MAIN OUTCOME MEASURES: Mortality for both operations according to surgeon. EuroSCORE to stratify patients into low and high risk. RESULTS: 10,163 patients underwent surgery under 25 surgeons. The average number of patients per surgeon was 363 for coronary artery surgery and 44 for aortic valve replacement. Seventeen per cent of the patients undergoing coronary artery surgery and half of those undergoing aortic valve surgery were considered high risk. The average mortality was 1.8% (range 0-3.8%) for coronary surgery and 1.9% (0-12.5%) for aortic valve surgery. Mortality for all surgeons fell below 99% control limits of the national mean for both operations. CONCLUSIONS: The presented mortality figures for the two cardiac operations fell within accepted limits for all surgeons. The division of outcomes according to low and high risk patients is imperfect but may help to inform the public about the complexities of this type of analysis and prevent surgeons avoiding high risk patents who may benefit from an operation. PMID- 15746132 TI - Uncertainty, consultation, and the context of medical care. PMID- 15746133 TI - Vertigo. PMID- 15746134 TI - Recent developments in vitamin D deficiency and muscle weakness among elderly people. PMID- 15746135 TI - Health promotion. PMID- 15746136 TI - What can the UK and US health systems learn from each other? PMID- 15746137 TI - A global health equity agenda for the G8 summit. PMID- 15746138 TI - Gastric rupture and laryngeal mask airway: negative "hype" this case report might generate is worrying. PMID- 15746139 TI - Independent medical school is long overdue. PMID- 15746140 TI - Electronic data protection: procedures need drastic improvement. PMID- 15746141 TI - Surgical management of metastatic inguinal lymphadenopathy: the term deep inguinal nodes should be abandoned. PMID- 15746142 TI - Gastric rupture and laryngeal mask airway: laryngeal mask airway was not likely cause of gastric rupture. PMID- 15746143 TI - Assessing drug safety. PMID- 15746144 TI - Surgical management of metastatic inguinal lymphadenopathy: sentinel node biopsy has cost implications for the NHS. PMID- 15746145 TI - Funding for GMC should come from taxpayers. PMID- 15746146 TI - Even pharmaceutical companies are vulnerable to loss of faith. PMID- 15746147 TI - Europe also needs agency for postmarketing surveillance. PMID- 15746148 TI - HER1/EGFR-targeted agents: predicting the future for patients with unpredictable outcomes to therapy. AB - Therapeutic agents that target the epidermal growth factor receptor (HER1/EGFR) signal pathway, such as small-molecule tyrosine kinase inhibitors and monoclonal antibodies are now advanced in clinical development and two are already licensed for use. Complete/ongoing phase II studies with these agents clearly demonstrate that a small, but significant proportion of patients respond to HER1/EGFR inhibition. However, with our current understanding of tumour biology and genetics, we cannot explain why some patients respond well and others less so or not at all. These differences may be a result of many factors, such as patients' genotype and phenotype, pharmacological and pharmacokinetic differences between agents or the inherent molecular heterogeneity of tumours. In this article, we explore current strategies to identify patients who respond differently and ways to maximise the clinical benefit of these therapies. This includes defining optimal dose and dosing schedules, identifying appropriate combination partners and finding predictive and surrogate markers of response. The association between HER1/EGFR gene mutations in non-small cell lung cancer (NSCLC) tumours and response to HER1/EGFR-targeted agents is also discussed. This may help us to preselect responsive patients, tailor the dose according to the individual's tolerability, or monitor these agents to optimise/interrupt therapy at an early stage. PMID- 15746149 TI - A unique exonic splice enhancer mutation in a family with X-linked mental retardation and epilepsy points to a novel role of the renin receptor. AB - The renin-angiotensin system (RAS) is essential for blood pressure control and water-electrolyte balance. Until the discovery of the renin receptor, renin was believed to be mainly a circulating enzyme with a unique function, the cleavage of angiotensinogen. We report a unique mutation in the renin receptor gene (ATP6AP2) present in patients with X-linked mental retardation and epilepsy (OMIM no. 300423), but absent in 1200 control X-chromosomes. A silent mutation (c.321C>T, p.D107D) residing in a putative exonic splicing enhancer site resulted in inefficient inclusion of exon 4 in 50% of renin receptor mRNA, as demonstrated by quantitative RT-PCR. Analysis of membrane associated-receptor molecular forms showed the presence of full-length and truncated proteins in the patient. Functional analysis demonstrated that the mutated receptor could bind renin and increase renin catalytic activity, similar to the wild-type receptor, but resulted in a modest and reproducible impairment of ERK1/2 activation. Thus, our findings confirm the importance of the RAS in cognitive processes and indicate a novel specific role for the renin receptor in cognitive functions and brain development. PMID- 15746150 TI - A functional single nucleotide polymorphism in the core promoter region of CALM1 is associated with hip osteoarthritis in Japanese. AB - Osteoarthritis (OA), a common skeletal disease, is a leading cause of disability among the elderly populations. OA is characterized by gradual loss of articular cartilage, but the etiology and pathogenesis of OA are largely unknown. Epidemiological and genetic studies have demonstrated that genetic factors play an important role in OA. To identify susceptibility genes for OA, we performed a large-scale, case-control association study using gene-based single nucleotide polymorphisms (SNPs). In two independent case-control populations, we found significant association (P=9.8x10(-7)) between hip OA and a SNP (IVS3-293C>T) located in intron 3 of the calmodulin (CaM) 1 gene (CALM1). CALM1 was expressed in cultured chondrocytes and articular cartilage, and its expression was increased in OA. Subsequent linkage-disequilibrium mapping identified five SNPs showing significant association equivalent to IVS3-293C>T. One of these (-16C>T) is located in the core promoter region of CALM1. Functional analyses indicate that the susceptibility -16T allele decreases CALM1 transcription in vitro and in vivo. Inhibition of CaM in chondrogenic cells reduced the expression of the major cartilage matrix genes Col2a1 and Agc1. These results suggest that the transcriptional level of CALM1 is associated with susceptibility for hip OA through modulation of chondrogenic activity. Our findings reveal the CALM1 mediated signaling pathway in chondrocytes as a novel potential target for treatment of OA. PMID- 15746151 TI - Frequent silencing of DBC1 is by genetic or epigenetic mechanisms in non-small cell lung cancers. AB - Genome-wide screening of DNA copy number aberrations in 27 cell lines derived from non-small cell lung cancers (NSCLCs), using a custom-made comparative genomic hybridization (CGH)-array, identified a homozygous deletion of the deleted in bladder cancer 1 gene (DBC1) in one cell line. Homozygous deletion of DBC1, located at 9q33.1, was also observed in two of 53 primary NSCLC tumors examined. Moreover, 21 of the other 26 cell lines showed complete loss of DBC1 expression, although normal lung tissues express this gene, and treatment with 5 aza-2'-deoxycytidine restored expression of DBC1. Hypermethylation in part of a CpG island around the exon 1 of DBC1 has been reported in urothelial cancers, but the potential association between methylation and expression status was never clarified in that disease. In our experiments, a different part of the same CpG island showed promoter activity in vitro and was frequently methylated in our cell lines and primary tumors of NSCLC, where methylation status correlated inversely with gene expression. Among our primary NSCLC cases, methylation of the DBC1 promoter occurred more frequently in men, elderly patients and smokers than in women, younger patients and nonsmokers respectively, but it was not correlated with tumor stage or histology. Exogenous overexpression of DBC1 in NSCLC cell lines lacking its expression inhibited cell growth. Our results provide the first evidence that DBC1 is a likely tumor suppressor for NSCLC; silencing of the gene through homozygous deletion or methylation of its promoter region may be associated with progression of this disease. PMID- 15746152 TI - Loci for regulation of bone mineral density in men and women identified by genome wide linkage scan: the FAMOS study. AB - Osteoporosis is a common disease with a strong genetic component, characterized by reduced bone mass and an increased risk of fracture. Bone mineral density (BMD) is a highly heritable trait and a key determinant of osteoporotic fracture risk, but the genes responsible are incompletely defined. Here, we identified quantitative trait loci (QTL) for regulation of BMD by a genome wide scan involving 3691 individuals from 715 families, who were selected because of reduced BMD values at the lumbar spine (LS-BMD) or femoral neck (FN-BMD) in probands. Linkage analysis was conducted in the study group as a whole with correction for age, gender, weight and height. Further analyses were conducted for men and women separately to identify gender-specific QTL and for those under and over the age of 50 years to distinguish QTL for peak bone mass from those that influence bone mass in older people. No regions of suggestive or significant linkage were identified when data from all subjects were analyzed together. On subgroup analysis, however, we identified a significant QTL for FN-BMD on chromosome 10q21 (LOD score +4.42; men < or =50 years) and two suggestive QTL for LS-BMD on chromosomes 18p11 (LOD score +2.83; women >50 years) and 20q13 (LOD score +3.20; women < or =50 years). We identified five other QTL for BMD with LOD scores of greater than +2.20 on chromosomes 3q25, 4q25, 7p14, 16p13 and 16q23. This study provides evidence for gender-specific, site-specific and age-specific QTL, which regulate BMD in humans, and illustrates the importance of conducting subgroup analysis to detect these loci. PMID- 15746153 TI - Inactivation of Rai1 in mice recapitulates phenotypes observed in chromosome engineered mouse models for Smith-Magenis syndrome. AB - Retinoic acid induced 1 (RAI1) is among the 20 genes identified in the critical region of Smith-Magenis syndrome (SMS), a genomic disorder with multiple congenital anomalies associated with a 3.7 Mb heterozygous deletion of 17p11.2. Heterozygous premature termination mutations in RAI1 have been identified recently in SMS patients without detectable deletions. To investigate Rai1 function, we generated a null allele in mice by gene targeting and simultaneously inserted a lacZ reporter gene into the Rai1 locus. X-gal staining of the Rai1(+/ ) mice recapitulated the endogenous expression pattern of Rai1. The gene was predominantly expressed in the epithelial cells involved in organogenesis. Obesity and craniofacial abnormalities, which have been reported in SMS mouse models containing a heterozygous deletion of the syntenic SMS critical region, were observed in Rai1(+/-) mice. Thus, haploinsufficiency of Rai1 causes obesity and craniofacial abnormalities in mice. Interestingly, the penetrance of craniofacial anomalies is further reduced in Rai1(+/-) mice. Most homozygous mice died during gastrulation and organogenesis. The surviving Rai1(-/-) mice were growth retarded and displayed malformations in both the craniofacial and the axial skeleton. Using green fluorescence protein and GAL4 DNA binding domain fusions to Rai1, we showed that Rai1 is translocated to the nucleus and it has transactivation activity. Our data are consistent with Rai1 functioning as a transcriptional regulator, document that Rai1 haploinsufficiency is responsible for obesity and craniofacial phenotypes in mice with SMS deletions, and indicate Rai1 is important for embryonic and postnatal developments. PMID- 15746154 TI - The influence of percutaneous atrial septal defect closure on the occurrence of migraine. AB - AIMS: Percutaneous patent foramen ovale closure seems to influence migraine. We wanted to observe the effect of percutaneous atrial septal defect (ASD) closure on migraine. METHODS AND RESULTS: All patients (>16 years of age) with a percutaneous ASD closure were selected from our database (n=114). A questionnaire about headache before and after closure was sent. According to the criteria of the International Headache Society, two neurologists diagnosed migraine with and without aura (MA+ and MA-, respectively). McNemar paired chi(2) and Wilcoxon signed rank tests were used where applicable. Seventy-five patients (66%, 59 females, mean age 51+/-19 years) responded and were included in the study. An Amplatzer ASD occluder was used in all. Median follow-up time was 29 months (IQ1 and IQ3, 18 and 39 months, respectively). The prevalence of MA- and MA+ changed from 19 (14/75) and 11% (8/75), respectively, before closure to 12 (9/75) and 15% (11/75), respectively, after closure (P=0.18 and P=0.55, respectively, vs. before closure). In 12 patients who suffered from migraine before closure (n=4 and 8, MA+ and MA-, respectively), migraine disappeared. In this subgroup, the frequency of migraine attacks decreased significantly (P=0.01). New-onset migraine was noted in 10 patients (n=7 and 3, MA+ and MA-, respectively). CONCLUSION: Percutaneous ASD closure was not related to a decrease in prevalence of migraine. In a subgroup, patients who suffered from typical migraine before ASD closure, the frequency of migraine attacks decreased significantly. The reason for the new onset migraine remains unexplained. A larger study sample will be necessary to determine these findings. PMID- 15746155 TI - TONSOKU is expressed in S phase of the cell cycle and its defect delays cell cycle progression in Arabidopsis. AB - TONSOKU(TSK)/MGOUN3/BRUSHY1 of Arabidopsis thaliana encodes a nuclear leucine glycine-aspargine (LGN) domain protein implicated to be involved in genome maintenance, and mutants with defects in TSK show a fasciated stem with disorganized meristem structures. We identified a homolog of TSK from tobacco BY 2 cells (NtTSK), which showed high sequence conservation both in the LGN domain and in leucine-rich repeats with AtTSK. The NtTSK gene was expressed during S phase of the cell cycle in tobacco BY-2 cells highly synchronized for cell division. The tsk mutants of Arabidopsis contained an increased proportion of cells with 4C nuclei and cells expressing cyclin B1 compared with the wild type. These results suggest that TSK is required during the cell cycle and defects of TSK cause the arrest of cell cycle progression at G2/M phase. PMID- 15746156 TI - Isolation and expression analysis of a tobacco AINTEGUMENTA ortholog (NtANTL). AB - The Arabidopsis AINTEGUMENTA (ANT) protein is essential for proper ovule development, but functions in cell proliferation and organ growth throughout the plant. Here we report the isolation of a full-length cDNA clone from tobacco (Nicotiana tabacum L.) that encodes a protein with high similarity to ANT and is preferentially expressed in the pistil. In situ hybridization analysis on the tobacco ovary shows that the expression pattern of the corresponding gene is different from that of ANT in Arabidopsis. PMID- 15746157 TI - Tissue localization of cytokinin dehydrogenase in maize: possible involvement of quinone species generated from plant phenolics by other enzymatic systems in the catalytic reaction. AB - The degradation of cytokinins in plants is controlled by the flavoprotein cytokinin dehydrogenase (EC 1.5.99.12). Cytokinin dehydrogenase from maize showed the ability to use oxidation products of guaiacol, 4-methylcatechol, acetosyringone and several other compounds as electron acceptors. These results led us to explore the cability for indirect production of suitable electron acceptors by different quinone-generating enzymes. The results reported here revealed that the electron acceptors may be generated in vivo from plant phenolics by other enzymatic systems such as peroxidase and tyrosinase/laccase/catechol oxidase. Histochemical localization of cytokinin dehydrogenase by activity staining and immunochemistry using optical and confocal microscopy showed that cytokinin dehydrogenase is most abundant in the aleurone layer of maize kernels and in phloem cells of the seedling shoots. Cytokinin dehydrogenase was confirmed to be present in the apoplast of cells. Co-staining of enzyme activity for laccase, an enzyme poised to function on the cell wall in the apoplast, in those tissues suggests a possible cooperation of the enzymes in cytokinin degradation. Additionally, the presence of precursors for electron acceptors of cytokinin dehydrogenase was detected in phloem exudates collected from maize seedlings, suggestive of an enzymatic capacity to control cytokinin flux through the vasculature. A putative metabolic connection between cytokinin degradation and conversion of plant phenolics by oxidases was proposed. PMID- 15746158 TI - Visualization of plastid nucleoids in situ using the PEND-GFP fusion protein. AB - Plastid DNA is a circular molecule of 120-150 kbp, which is organized into a protein-DNA complex called a nucleoid. Although various plastids other than chloroplasts exist, such as etioplasts, amyloplasts and chromoplasts, it is not easy to observe plastid nucleoids within the cells of many non-green tissues. The PEND (plastid envelope DNA-binding) protein is a DNA-binding protein in the inner envelope membrane of developing chloroplasts, and a DNA-binding domain called cbZIP is present at its N-terminus. We made various PEND-green fluorescent protein (GFP) fusion proteins using the cbZIP domains from various plants, and found that they were localized in the chloroplast nucleoids in transient expression in leaf protoplasts. In stable transformants of Arabidopsis thaliana, PEND-GFP fusion proteins were also localized in the nucleoids of various plastids. We have succeeded in visualizing plastid nucleoids in various intact tissues using this stable transformant. This technique is useful in root, flower and pollen, in which it had been difficult to observe plastid nucleoids. The relative arrangement of nucleoids within a chloroplast was kept unchanged when the chloroplast moved within a cell. During the division of plastid, nucleoids formed a network structure, which made possible equal partition of nucleoids. PMID- 15746159 TI - Tumour regulation of fibroblast hyaluronan expression: a mechanism to facilitate tumour growth and invasion. AB - Hyaluronan, a high molecular weight glycosaminoglycan is associated with cellular proliferation and migration. In a number of different tumour types, there is a close correlation between tumour progression and hyaluronan production, either by the tumour cells or the surrounding stromal cells. We have examined the ability of an aggressive melanoma cell line (C8161) to stimulate the synthesis of fibroblast hyaluronan, and the association of cell-surface CD44 receptors and hyaluronan with invasion. Melanoma cell-conditioned medium (CM) prepared in low glucose medium (1 mg/ml) stimulated the synthesis of fibroblast glycosaminoglycan as measured by [3H] glucosamine incorporation, and the synthesis of hyaluronan as measured using a specific hyaluronan-binding plate assay, while tumour cell-CM prepared in high glucose medium (4.5 mg/ml) inhibited the synthesis of fibroblast glycosaminoglycan. High glucose tumour cell-CM contained large amounts of lactate that appeared to inhibit the tumour-derived factor stimulation of fibroblast glycosaminoglycan synthesis, as removal of the lactate restored the stimulating activity. Melanoma cells seeded on contracted collagen lattices and incubated at the air/liquid interface rapidly formed a multilayered cell mass on the surface, with significant invasion of the gel. Hyaluronan staining was apparent within the collagen gel, and strong staining was seen around the invading tumour cells, but not around those cell layers near the surface. CD44 expression on the tumour cells was confined to those invading cells and corresponded to cellular hyaluronan staining. Hyaluronan staining was also apparent around and between tumour cells invading fibroblast-free collagen lattices. Monolayer cultures of C8161 cells stained strongly for CD44, but few cells stained for hyaluronan, while no detectable hyaluronan was released into the medium. In summary, the C8161 melanoma cells stimulated the synthesis of fibroblast hyaluronan, and in collagen lattices, only the invasive tumour cells expressed CD44 and hyaluronan, either in the presence or absence of fibroblasts. PMID- 15746160 TI - Constitutional short telomeres are strong genetic susceptibility markers for bladder cancer. AB - Lack of functional telomeres can cause chromosomal aberrations. This type of genetic instability may promote tumorigenesis. We have investigated the association between mean telomere length in buccal cells (assessed with quantitative real-time PCR) and bladder cancer risk in a case-control study. Patients with bladder cancer displayed significantly shorter telomeres than control subjects (P = 0.001). Median telomere length ratio was 0.95 (range 0.53 3.2) for cases and 1.1 (0.51-2.4) for controls. Moreover, the adjusted odds ratio (OR) for bladder cancer was significantly increased in the quartile with the shortest telomere length OR = 4.5 [95% confidence interval (CI) 1.7-12]. It is known that oxidative stress, alkylation or UV radiation increases shortening of telomeres. Therefore, we also analyzed whether environmental and genetic factors associated with DNA damage, i.e. smoking and polymorphisms in the genes involved in the metabolism of genotoxic carcinogens (EPHX1, GSTA1, GSTM1, GSTP1, GSTT1, NAT1, NAT2 and NQO1) or DNA repair (APE1, NBS1, XPC, XPD, XRCC1, XRCC3 and XRCC4), could modify the association between telomere length and cancer risk. A clear effect of smoking and telomere length could be observed. Current smokers with short telomeres had more than six times as higher risk as non-smokers/former smokers with long telomeres (OR = 6.3, 95% CI 1.7-23). Lack of the biotransformation gene GSTM1 and short telomeres were associated with OR = 6.5 (95% CI 2.4-18), whereas homozygous carriers of 312Asn in the DNA repair gene XPD, with short telomeres, displayed an OR of 17 (95% CI 1.9-150). However, no significant interaction for cancer risk could be proven for telomere length, smoking and susceptibility genotypes of metabolizing and DNA-repairing genes. PMID- 15746161 TI - Bowman-Birk inhibitor abates proteasome function and suppresses the proliferation of MCF7 breast cancer cells through accumulation of MAP kinase phosphatase-1. AB - The Bowman-Birk inhibitor (BBI), a soybean-derived protease inhibitor with well characterized ability to inhibit trypsin and chymotrypsin activities, has been shown to be an effective suppressor of carcinogenesis and treated in human phase IIa clinical trial. However, the precise mechanisms by which BBI suppresses carcinogenesis are unknown. In this study, we demonstrated that BBI specifically and potently inhibits the proteasomal chymotrypsin-like activity in vitro and in vivo in MCF7 breast cancer cells. Proteasome inhibition by BBI is associated with accumulation of ubiquitinated proteins and the proteasome substrates, p21Cip1/WAF1 and p27Kip1, accompanied with downregulation of cyclin D1 and cyclin E which could arrest cell cycle at G1/S phase. Moreover, BBI suppressed MCF7 cell growth and had a novel effect on the decrease of phosphorylated extracellular signal-related kinases (ERK1/2). However, BBI was unable to inactivate ERK1/2 in the presence of a phosphatase inhibitor or a transcription inhibitor suggesting the involvement of a specific phosphatase. We found an induction of MAP kinase phosphatase-1 (MKP-1) in dose- and time-dependent manner correlated with dephosphorylation of ERK1/2 in BBI-treated MCF7 cells. In addition, BBI exhibited no inhibitory effects on EGF-stimulated activation of ERK1/2 and Akt. Together, we suggested that BBI abates proteasome function and results in upregulation of MKP-1, which in turn suppresses ERK1/2 activity. Our results support the notion that proteasome inhibition by BBI is a novel mechanism that contributes to prevention of cancer and further provides evidence that soybean products have the potential to advance as chemopreventive agents. PMID- 15746162 TI - Cytokines differentially regulate the synthesis of prostanoid and nitric oxide mediators in tumorigenic versus non-tumorigenic mouse lung epithelial cell lines. AB - Studies using transgenic and knockout mice have demonstrated that particular cytokines influence lung tumor growth and identified prostaglandin E2 (PGE2), prostacyclin (PGI2) and nitric oxide (NO) as critical mediators of this process. PGE2 and NO were pro-tumorigenic while PGI2 was antitumorigenic. We describe herein an in vitro experimental approach to examine interactions among cytokines, prostaglandins (PGs) and NO. PGE2, PGI2, and NO levels were assayed in culture media from non-tumorigenic mouse lung epithelial cell lines, their spontaneous transformants and mouse lung tumor-derived cell lines, before or after exposure to the cytokines TNFalpha, IFNgamma and IL1beta, alone and in combination. More PGE2 than PGI2 was produced by neoplastic cells, while the opposite was observed in non-tumorigenic lines. Cytokine exposure magnified the extent of these differential concentrations. The PGE2 to PGI2 ratio was also greater in chemically-induced mouse lung tumors than in adjacent tissue or control lungs, supporting the physiological relevance of this in vitro model. Expression of PG biosynthetic enzymes in these cell lines correlated with production of the corresponding PGs. Cytokine treatment enhanced NO production by inducing the inflammation-associated biosynthetic enzyme, inducible NO synthase (iNOS), but this did not correlate with the neoplastic status of cells. Inhibition of iNOS or cyclooxygenase 2 activity using aminoguanidine or NS-398 respectively, demonstrated that NO did not affect PG production nor did PGs influence NO production. Since lack of iNOS inhibits mouse lung tumor formation, we propose that this is independent of any modulation of PG synthesis in epithelial cells. The similar normal/neoplastic trends in PGE2 to PGI2 ratios both in vitro and in vivo, together with an amplification of this difference upon cytokine exposure, are consistent with the hypothesis that cytokines released during inflammation exacerbate differences in the behavior of neoplastic and normal lung cells. PMID- 15746163 TI - Butyrate may enhance toxicological defence in primary, adenoma and tumor human colon cells by favourably modulating expression of glutathione S-transferases genes, an approach in nutrigenomics. AB - Butyrate, formed by bacterial fermentation of plant foods, has been suggested to reduce colon cancer risks by suppressing the proliferation of tumor cells. In addition, butyrate has been shown to induce glutathione S-transferases (GSTs) in tumor cell lines, which may contribute to the detoxification of dietary carcinogens. We hypothesize that butyrate also affects biotransformation in non transformed colon cells. Thus, we have investigated the gene expression of drug metabolism genes in primary human colon tissue, premalignant LT97 adenoma and HT29 tumor cells cultured in an appropriate medium+/-butyrate. A total of 96 drug metabolism genes (including 12 GSTs) spotted on cDNA macroarrays (Superarray; n = 3) were hybridized with biotin-labeled cDNA probes. To validate the expression detected with Superarray, samples of LT97 cells were also analyzed with high density microarrays (Affymetrix U133A), which include biotransformation genes that overlap with the set of genes represented on the Superarray. Relative expression levels were compared across colon samples and for each colon sample+/ butyrate. Compared with fresh tissue, 13 genes were downregulated in primary cells cultivated ex vivo, whereas 8 genes were upregulated. Several genes were less expressed in LT97 (40 genes) or in HT29 (41 and 17 genes, grown for 72 and 48 h, respectively) compared with primary colon tissue. Butyrate induced GSTP1, GSTM2, and GSTA4 in HT29 as previously confirmed by other methods (northern blot/qPCR). We detected an upregulation of GSTs (GSTA2, GSTT2) that are known to be involved in the defence against oxidative stress in primary cells upon incubation with butyrate. The changes in expression detected in LT97 by Superarray and Affymetrix were similar, confirming the validity of the results. We conclude that low GST expression levels were favourably altered by butyrate. An induction of the toxicological defence system possibly contributes to reported chemopreventive properties of butyrate, a product of dietary fibre fermentation in the gut. PMID- 15746164 TI - Prevention of mouse skin tumor promotion by dietary energy restriction requires an intact adrenal gland and glucocorticoid supplementation restores inhibition. AB - Our laboratory has demonstrated in the previous studies that dietary energy restriction (DER) inhibited the promotion of skin tumorigenesis and others have found that adrenalectomy may reverse that inhibition. The purpose of the research reported here was to determine if circulating corticosterone (CCS) may be the adrenal hormone responsible for DER prevention of skin carcinogenesis. Female SENCAR mice were initiated with 7,12-dimethylbenzanthracene (DMBA) and promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA) in either sham-operated or adrenalectomized (ADX) mice fed ad libitum (AL) or energy restricted diets. DER was 60% of the AL calorie intake with the removal of energy from fat and carbohydrate. CCS, the main glucocorticoid hormone secreted by the murine adrenal gland, was added to the drinking water of AL/ADX and DER/ADX groups to determine the role of CCS in the DER inhibition of tumor development. In sham-operated groups, DER compared with AL-fed mice experienced significantly decreased papilloma incidence and multiplicity (P < 0.0001). ADX did not alter papilloma incidence or multiplicity in AL-fed mice but ADX partially reversed the inhibition of papilloma multiplicity and incidence in DER mice. CCS supplementation to both DER/ADX and AL/ADX mice resulted in reduced papilloma incidence and multiplicity. In DER/ADX mice, CCS dramatically reduced papilloma rates while in AL/ADX mice CCS reduced the papilloma rates to those seen in the DER sham group. DER significantly reduced carcinoma multiplicity mean counts per effective animal (P < 0.0001) compared with AL-fed groups in sham and ADX/CCS groups. DER/ADX mice lost the carcinoma multiplicity protection seen in sham/DER mice. CCS treatment of ADX mice significantly decreased total carcinoma (in situ and invasive) incidence rates per effective animal (P < 0.0003). ADX followed by CCS treatment in the DER mice resulted in the lowest carcinoma incidence and multiplicity. Thus, DER-inhibition of skin tumorigenesis was mediated at least in part through CCS. However, CCS was more effective in preventing papillomas and carcinomas in DER/ADX mice than in AL/ADX mice, suggesting that other factors may also be involved in the DER prevention of tumor formation. PMID- 15746165 TI - Prior heavy exercise eliminates VO2 slow component and reduces efficiency during submaximal exercise in humans. AB - We investigated the hypothesis that the pulmonary oxygen uptake (VO2) slow component is related to a progressive increase in muscle lactate concentration and that prior heavy exercise (PHE) with pronounced acidosis alters VO2 kinetics and reduces work efficiency. Subjects (n= 9) cycled at 75% of the peak VO2 (VO2peak) for 10 min before (CON) and after (AC) PHE. VO2 was measured continuously (breath-by-breath) and muscle biopsies were obtained prior to and after 3 and 10 min of exercise. Muscle lactate concentration was stable between 3 and 10 min of exercise but was 2- to 3-fold higher during AC (P < 0.05 versus CON). Acetylcarnitine (ACn) concentration was 6-fold higher prior to AC and remained higher during exercise. Phosphocreatine (PCr) concentration was similar prior to exercise but the decrease was 2-fold greater during AC than during CON. The time constant for the initial VO2 kinetics (phase II) was similar but the asymptote was 14% higher during AC. The slow increase in VO2 between 3 and 10 min of exercise during CON (+7.9 +/- 0.2%) was not correlated with muscle or blood lactate levels. PHE eliminated the slow increase in VO2 and reduced gross exercise efficiency during AC. It is concluded that the VO2 slow component cannot be explained by a progressive acidosis because both muscle and blood lactate levels remained stable during CON. We suggest that both the VO2 slow component during CON and the reduced gross efficiency during AC are related to impaired contractility of the working fibres and the necessity to recruit additional motor units. Despite a pronounced stockpiling of ACn during AC, initial VO2 kinetics were not affected by PHE and PCr concentration decreased to a lower plateau. The discrepancy with previous studies, where initial oxidative ATP generation appears to be limited by acetyl group availability, might relate to remaining fatiguing effects of PHE. PMID- 15746166 TI - In vivo recordings from rat geniculate ganglia: taste response properties of individual greater superficial petrosal and chorda tympani neurones. AB - Coding of gustatory information is complex and unique among sensory systems; information is received by multiple receptor populations located throughout the oral cavity and carried to a single central relay by four separate nerves. The geniculate ganglion is the location of the somata of two of these nerves, the greater superficial petrosal (GSP) and the chorda tympani (CT). The GSP innervates taste buds on the palate and the CT innervates taste buds on the anterior tongue. To obtain requisite taste response profiles of GSP neurones, we recorded neurophysiological responses to taste stimuli of individual geniculate ganglion neurones in vivo in the rat and compared them to those from the CT. GSP neurones had a distinct pattern of responding compared to CT neurones. For example, a small subset of GSP neurones had high response frequencies to sucrose stimulation, whereas no CT neurones had high response frequencies to sucrose. In contrast, NaCl elicited high response frequencies in a small subset of CT neurones and elicited moderate response frequencies in a relatively large proportion of GSP neurones. The robust whole-nerve response to sucrose in the GSP may be attributable to relatively few, narrowly tuned neurones, whereas the response to NaCl in the GSP may relate to proportionately more, widely tuned neurones. These results demonstrate the diversity in the initial stages of sensory coding for two separate gustatory nerves involved in the ingestion or rejection of taste solutions, and may have implications for central coding of gustatory quality and concentration as well as coding of information used in controlling energy, fluid and electrolyte homeostasis. PMID- 15746167 TI - After-effects of near-threshold stimulation in single human motor axons. AB - Subthreshold electrical stimuli can generate a long-lasting increase in axonal excitability, superficially resembling the phase of superexcitability that follows a conditioning nerve impulse. This phenomenon of 'subthreshold superexcitability' has been investigated in single motor axons in six healthy human subjects, by tracking the excitability changes produced by conditioning stimuli of different amplitudes and waveforms. Near-threshold 1 ms stimuli caused a mean decrease in threshold at 5 ms of 22.1 +/- 6.0% (mean +/-s.d.) if excitation occurred, or 6.9 +/- 2.6% if excitation did not occur. The subthreshold superexcitability was maximal at an interval of about 5 ms, and fell to zero at 30 ms. It appeared to be made up of two components: a passive component linearly related to conditioning stimulus amplitude, and a non-linear active component. The active component appeared when conditioning stimuli exceeded 60% of threshold, and accounted for a maximal threshold decrease of 2.6 +/- 1.3%. The passive component was directly proportional to stimulus charge, when conditioning stimulus duration was varied between 0.2 and 2 ms, and could be eliminated by using triphasic stimuli with zero net charge. This change in stimulus waveform had little effect on the active component of subthreshold superexcitability or on the 'suprathreshold superexcitability' that followed excitation. It is concluded that subthreshold superexcitability in human motor axons is mainly due to the passive electrotonic effects of the stimulating current, but this is supplemented by an active component (about 12% of suprathreshold superexcitability), due to a local response of voltage-dependent sodium channels. PMID- 15746168 TI - Physiological properties of rod photoreceptor electrical coupling in the tiger salamander retina. AB - Using dual whole-cell voltage and current clamp recording techniques, we investigated the gap junctional conductance and the coupling coefficient between neighbouring rods in live salamander retinal slices. The application of sinusoidal stimuli over a wide range of temporal frequencies allowed us to characterize the band-pass filtering properties of the rod network. We found that the electrical coupling of all neighbouring rods exhibited reciprocal and symmetrical conductivities. On average, the junctional conductance between paired rods was 500 pS and the coupling coefficient (the ratio of voltage responses of the follower cell to those of the driver cell), or K-value, was 0.07. Our experimental results also demonstrated that the rod network behaved like a band pass filter with a peak frequency of about 2-5 Hz. However, the gap junctions between adjacent rods exhibited linearity and voltage independency within the physiological range of rods. These gap junctions did not contribute to the filtering mechanisms of the rod network. Combined with the computational modelling, our data suggest that the filtering of higher frequency rod signals by the network is largely mediated by the passive resistive and capacitive (RC) properties of rod plasma membranes. Furthermore, we found several attributes of rod electrical coupling resembling the physiological properties of gene-encoded Cx35/36 gap junctions examined in other in vitro studies. This indicates that the previously found Cx35/36 expression in the salamander rod network may be functionally involved in rod-rod electrical coupling. PMID- 15746169 TI - The vagus regulates histamine mobilization from rat stomach ECL cells by controlling their sensitivity to gastrin. AB - The ECL cells in the oxyntic mucosa secrete histamine in response to gastrin, stimulating parietal cells to produce acid. Do they also operate under nervous control? The present study examines histamine mobilization from rat stomach ECL cells in situ in response to acute vagal excitation and to food or gastrin following vagal or sympathetic denervation. Applying the technique of microdialysis, we monitored the release of histamine by radioimmunoassay. Microdialysis probes were placed in the submucosa on either side of the stomach, 3 days before experiments. The rats were awake during microdialysis except when subjected to electrical vagal stimulation. One-sided electrical vagal stimulation raised serum gastrin and mobilized gastric histamine. However, gastrin receptor blockade prevented the histamine mobilization, indicating that circulating gastrin accounts for the response. Vagal excitation by hypoglycaemia (insulin) or pylorus ligation did not mobilize either gastrin or histamine. The histamine response to food was almost abolished by gastrin receptor blockade, and it was halved on the denervated side after unilateral subdiaphragmatic vagotomy. While the histamine response to a near-maximally effective dose of gastrin was unaffected by vagotomy, the response to low gastrin doses was reduced significantly. Abdominal ganglionic sympathectomy failed to affect the histamine response to either food or gastrin. In conclusion, gastrin is responsible for most of the food-evoked mobilization of ECL-cell histamine. The histamine response to electrical vagal stimulation reflects the effect of circulating gastrin rather than a direct action of the vagus on the ECL cells. Vagal denervation was accompanied by an impaired histamine response to food intake, probably reflecting the right-ward shift of the serum gastrin concentration histamine response curve. The results suggest that the vagus controls the sensitivity of the ECL cells to gastrin. PMID- 15746170 TI - Phase-dependent and task-dependent modulation of stretch reflexes during rhythmical hand tasks in humans. AB - Phase-dependent and task-dependent modulation of reflexes has been extensively demonstrated in leg muscles during locomotory activity. In contrast, the modulation of reflex responses of hand muscles during rhythmic movement is poorly documented. The objective of this study was to determine whether comparable reflex modulation occurs in muscles controlling finger motions during rhythmic, fine-motor tasks akin to handwriting. Twelve healthy subjects performed two rhythmic tasks while reflexes were evoked by mechanical perturbations applied at various phases of each task. Electromyograms (EMGs) were recorded from four hand muscles, and reflexes were averaged during each task relative to the movement phase. Stretch reflexes in all four muscles were found to be modulated in amplitude with respect to the phase of the rhythmic tasks, and also to vary distinctly with the tasks being conducted. The extent and pattern of reflex modulation differed between muscles in the same task, and between tasks for the same muscle. Muscles with a primary role in each task showed a higher correlation between reflex response and background EMG than other muscles. The results suggest that the modulation patterns observed may reflect optimal strategies of central-peripheral interactions in controlling the performance of fine-motor tasks. As with comparable studies on locomotion, the phase-dependency of the stretch reflexes implies a dynamically fluctuating role of proprioceptive feedback in the control of the hand muscles. The clear task-dependency is also consistent with a dynamic interaction of sensory feedback and central programming, presumably adapted to facilitate the successful performance of the different fine-motor tasks. PMID- 15746171 TI - Disruption of excitation-contraction coupling and titin by endogenous Ca2+ activated proteases in toad muscle fibres. AB - This study investigated the effects of elevated, physiological levels of intracellular free [Ca(2+)] on depolarization-induced force responses, and on passive and active force production by the contractile apparatus in mechanically skinned fibres of toad iliofibularis muscle. Excitation-contraction (EC) coupling was retained after skinning and force responses could be elicited by depolarization of the transverse-tubular (T-) system. Raising the cytoplasmic [Ca(2+)] to approximately 1 microm or above for 3 min caused an irreversible reduction in the depolarization-induced force response by interrupting the coupling between the voltage sensors in the T-system and the Ca(2+) release channels in the sarcoplasmic reticulum. This uncoupling showed a steep [Ca(2+)] dependency, with 50% uncoupling at approximately 1.9 microm Ca(2+). The uncoupling occurring with 2 microm Ca(2+) was largely prevented by the calpain inhibitor leupeptin (1 mm). Raising the cytoplasmic [Ca(2+)] above 1 microm also caused an irreversible decline in passive force production in stretched skinned fibres in a manner graded by [Ca(2+)], though at a much slower relative rate than loss of coupling. The progressive loss of passive force could be rapidly stopped by lowering [Ca(2+)] to 10 nm, and was almost completely inhibited by 1 mm leupeptin but not by 10 microm calpastatin. Muscle homogenates preactivated by Ca(2+) exposure also evidently contained a diffusible factor that caused damage to passive force production in a Ca(2+)-dependent manner. Western blotting showed that: (a) calpain-3 was present in the skinned fibres and was activated by the Ca(2+)exposure, and (b) the Ca(2+) exposure in stretched skinned fibres resulted in proteolysis of titin. We conclude that the disruption of EC coupling occurring at elevated levels of [Ca(2+)] is likely to be caused at least in part by Ca(2+) activated proteases, most likely by calpain-3, though a role of calpain-1 is not excluded. PMID- 15746173 TI - Expression pattern of neuronal and skeletal muscle voltage-gated Na+ channels in the developing mouse heart. AB - In the mammalian heart, a variety of voltage-gated Na(+) channel transcripts and proteins have been detected. However, little quantitative information is available on the abundance of each transcript during development, or the contribution of TTX-sensitive Na(+) channels to the cardiac sodium current (I(Na)). Using competitive and real-time RT-PCR we investigated the transcription of six Na(+) channels (Na(v)1.1-Na(v)1.6) and the beta1 subunit during mouse heart development. Na(v)1.5 was predominantly expressed in the adult heart, whereas the splice variant Na(v)1.5a was the major Na(+) channel isoform in embryonic hearts. The TTX-resistant Na(+) channel transcripts (Na(v)1.5 and Na(v)1.5a) increased 1.7-fold during postnatal development. Transcripts encoding TTX-sensitive Na(+) channels (Na(v)1.1-Na(v)1.4) and the beta1 subunit gradually increased up to fourfold from postnatal day (P)1 to P126, while the Na(v)1.6 transcript level remained low and constant over the same period. In adults, TTX sensitive channel mRNA accounted for 30-40% of the channel pool in whole-heart preparations (Na(v)1.3 > Na(v)1.4 > Na(v)1.2 >> Na(v)1.1 approximately Na(v)1.6), and 16% in mRNA from isolated cardiomyocytes (Na(v)1.4 > Na(v)1.3 > Na(v)1.2 > Na(v)1.1 > Na(v)1.6). Confocal immunofluorescence on ventricular myocytes suggested that Na(v)1.1 and Na(v)1.2 were localized at the intercalated disks and in the t tubules. Na(v)1.3 labelling predominantly produced a diffuse but strong intracellular signal. Na(v)1.6 fluorescence was detected only along the Z lines. Electrophysiological recordings showed that TTX-sensitive and TTX-resistant Na(+) channels, respectively, accounted for 8% and 92% of the I(Na) in adult ventricular cardiomyocytes. Our data suggest that neuronal and skeletal muscle Na(+) channels contribute to the action potential of cardiomyocytes in the adult mammalian heart. PMID- 15746172 TI - Modulation of skeletal and cardiac voltage-gated sodium channels by calmodulin. AB - Calmodulin (CaM) has been shown to modulate different ion channels, including voltage-gated sodium channels (NaChs). Using the yeast two-hybrid assay, we found an interaction between CaM and the C-terminal domains of adult skeletal (NaV1.4) and cardiac (NaV1.5) muscle NaChs. Effects of CaM were studied using sodium channels transiently expressed in CHO cells. Wild type CaM (CaM(WT)) caused a hyperpolarizing shift in the voltage dependence of activation and inactivation for NaV1.4 and activation for NaV1.5. Intracellular application of CaM caused hyperpolarizing shifts equivalent to those seen with CaM(WT) coexpression with NaV1.4. Elevated Ca2+ and CaM-binding peptides caused depolarizing shifts in the inactivation curves seen with CaM(WT) coexpression with NaV1.4. KN93, a CaM kinase II inhibitor, had no effect on NaV1.4, suggesting that CaM acts directly on NaV1.4 and not through activation of CaM-kinase II. Coexpression of hemi mutant CaMs showed that an intact N-terminal lobe of CaM is required for effects of CaM upon NaV1.4. Mutations in the sodium channel IQ domain disrupted the effects of CaM on NaV1.4: the I1727E mutation completely blocked all calmodulin effects, while the L1736R mutation disrupted the effects of Ca2+-calmodulin on inactivation. Chimeric channels of NaV1.4 and NaV1.5 also indicated that the C terminal domain is largely responsible for CaM effects on inactivation. CaM had little effect on NaV1.4 expressed in HEK cells, possibly due to large differences in the endogenous expression of beta-subunits between CHO and HEK cells. These results in heterologous cells suggest that Ca2+ released during muscle contraction rapidly modulates NaCh availability via CaM. PMID- 15746175 TI - Nanomedicine: current status and future prospects. AB - Applications of nanotechnology for treatment, diagnosis, monitoring, and control of biological systems has recently been referred to as "nanomedicine" by the National Institutes of Health. Research into the rational delivery and targeting of pharmaceutical, therapeutic, and diagnostic agents is at the forefront of projects in nanomedicine. These involve the identification of precise targets (cells and receptors) related to specific clinical conditions and choice of the appropriate nanocarriers to achieve the required responses while minimizing the side effects. Mononuclear phagocytes, dendritic cells, endothelial cells, and cancers (tumor cells, as well as tumor neovasculature) are key targets. Today, nanotechnology and nanoscience approaches to particle design and formulation are beginning to expand the market for many drugs and are forming the basis for a highly profitable niche within the industry, but some predicted benefits are hyped. This article will highlight rational approaches in design and surface engineering of nanoscale vehicles and entities for site-specific drug delivery and medical imaging after parenteral administration. Potential pitfalls or side effects associated with nanoparticles are also discussed. PMID- 15746174 TI - CFTR fails to inhibit the epithelial sodium channel ENaC expressed in Xenopus laevis oocytes. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) plays a crucial role in regulating fluid secretion by the airways, intestines, sweat glands and other epithelial tissues. It is well established that the CFTR is a cAMP activated, nucleotide-dependent anion channel, but additional functions are often attributed to it, including regulation of the epithelial sodium channel (ENaC). The absence of CFTR-dependent ENaC inhibition and the resulting sodium hyperabsorption were postulated to be a major electrolyte transport abnormality in cystic fibrosis (CF)-affected epithelia. Several ex vivo studies, including those that used the Xenopus oocyte expression system, have reported ENaC inhibition by activated CFTR, but contradictory results have also been obtained. Because CFTR-ENaC interactions have important implications in the pathogenesis of CF, the present investigation was undertaken by our three independent laboratories to resolve whether CFTR regulates ENaC in oocytes and to clarify potential sources of previously reported dissimilar observations. Using different experimental protocols and a wide range of channel expression levels, we found no evidence that activated CFTR regulates ENaC when oocyte membrane potential was carefully clamped. We determined that an apparent CFTR-dependent ENaC inhibition could be observed when resistance in series with the oocyte membrane was not low enough or the feedback voltage gain was not high enough. We suggest that the inhibitory effect of CFTR on ENaC reported in some earlier oocyte studies could be attributed to problems arising from high levels of channel expression and suboptimal recording conditions, that is, large series resistance and/or insufficient feedback voltage gain. PMID- 15746176 TI - Antibody phage display technologies with special reference to angiogenesis. AB - The presence of blood vessels is a prerequisite for normal development, tissue growth, and tissue repair. However, its abnormal occurrence or absence can also potentiate disease processes. Angiogenic therapies have been used to stimulate blood vessel growth in ischemic conditions such as severe end-stage peripheral vascular disease, ischemic heart disease and stroke and for inhibition of angiogenesis in tumors. The targeting and identification of novel endothelial cell (EC) markers that can ultimately be used in angiogenic strategies is an expanding field but is limited by the availability of reagents. For instance repeated injection of mouse monoclonal antibodies (Mabs) against angiogenic EC, can result in the production of autoantibodies. Therefore, these mouse Mabs cannot be used for therapeutic purposes. Phage display technology was employed in this context to select antibodies, proteins, and peptides against known or novel EC antigens. Furthermore, technologies have been developed that enable the specific targeting of epitopes on cells including the endothelium with high affinity/avidity antibodies. The focus for these antibody targeting strategies are markers that are unique or up-regulated on angiogenic EC including the vascular endothelial growth factor receptor (VEGFR) KDR, endoglin (CD105), and the extracellular domain B (ED-B) domain of fibronectin (FN). These markers are reviewed herein. PMID- 15746177 TI - Apigenin inhibits VEGF and HIF-1 expression via PI3K/AKT/p70S6K1 and HDM2/p53 pathways. AB - Apigenin is a nontoxic dietary flavonoid that has been shown to possess anti tumor properties and therefore poses special interest for the development of a novel chemopreventive and/or chemotherapeutic agent for cancer. Ovarian cancer is one of the most common causes of cancer death among women. Here we demonstrate that apigenin inhibits expression of vascular endothelial growth factor (VEGF) in human ovarian cancer cells. VEGF plays an important role in tumor angiogenesis and growth. We found that apigenin inhibited VEGF expression at the transcriptional level through expression of hypoxia-inducible factor 1alpha (HIF 1alpha). Apigenin inhibited expression of HIF-1alpha and VEGF via the PI3K/AKT/p70S6K1 and HDM2/p53 pathways. Apigenin inhibited tube formation in vitro by endothelial cells. These findings reveal a novel role of apigenin in inhibiting HIF-1 and VEGF expression that is important for tumor angiogenesis and growth, identifying new signaling molecules that mediate this regulation. PMID- 15746178 TI - Brief hypoxia before normoxic reperfusion (postconditioning) protects the heart against ischemia-reperfusion injury by preventing mitochondria peroxyde production and glutathione depletion. AB - Several recent works have shown that a brief ischemia applied during the onset of reperfusion (postconditioning) is cardioprotective in different animal models and that the early minutes of reperfusion are critical to its cardioprotection. This effect has been related to prevention of oxidative stress, but mechanisms have not been clearly demonstrated. The present study tested the hypothesis that mitochondria play a central role in peroxide production and oxidative stress during reperfusion and are responsible for the protective effect of postconditioning. Isolated perfused rat hearts were subjected to complete global ischemia for 45 min and reperfused for 40 min. Normoxic group was reperfused with a Krebs-Henseleit solution with the preischemic pO2 level (600 mmHg); in the "hypoxic group," normoxic reperfusion was preceded by 3 min with 150 mmHg pO2. Reperfusion was stopped at 3 and 40 min. The rate of hydroperoxide production, GSH, GSSG, and carbonyl protein levels were measured in mitochondria at 3 min and at the end of reperfusion. GSH and GSSG were also measured in tissue. Hemodinamic function was monitored during the experiment. LVEDp increased and LVDp decreased in the normoxic group but not in the hypoxic group. The rate of mitochondrial peroxide production was higher in normoxic than in the hypoxic group 3 min after reperfusion and at its conclusion. Accordingly, GSH was oxidized in normoxic but not in hypoxic hearts. Mitochondria carbonyl proteins were significantly higher in normoxic than in the hypoxic group at the end of reperfusion. In this model, 1) hypoxic reperfusion at the onset of reperfusion reduces myocardial injury; 2) the major rate of mitochondrial peroxide production is 3 min after the onset of reperfusion; 3) cardioprotection of postconditioning correlates with reduced mitochondria peroxide production and prevention of GSH oxidation. PMID- 15746179 TI - TNF-alpha acts via p38 MAPK to stimulate expression of the ubiquitin ligase atrogin1/MAFbx in skeletal muscle. AB - Atrogin1/MAFbx is an ubiquitin ligase that mediates muscle atrophy in a variety of catabolic states. We recently found that H2O2 stimulates atrogin1/MAFbx gene expression. Since the cytokine tumor necrosis factor-alpha (TNF-alpha) stimulates both reactive oxygen production and general activity of the ubiquitin conjugating pathway, we hypothesized that TNF-alpha would also increase atrogin1/MAFbx gene expression. As with H2O2, we found that TNF-alpha exposure up-regulates atrogin1/MAFbx mRNA within 2 h in C2C12 myotubes. Intraperitoneal injection of TNF-alpha increased atrogin1/MAFbx mRNA in skeletal muscle of adult mice within 4 h. Exposing myotubes to either TNF-alpha or H2O2 also produced general activation of the mitogen-activated protein kinases (MAPKs): p38, ERK1/2, and JNK. The increase in atrogin1/MAFbx gene expression induced by TNF-alpha was not altered significantly by ERK inhibitor PD98059 or the JNK inhibitor SP600125. In contrast, atrogin1/MAFbx up-regulation and the associated increase in ubiquitin conjugating activity were both blunted by p38 inhibitors, either SB203580 or curcumin. These data suggest that TNF-alpha acts via p38 to increase atrogin1/MAFbx gene expression in skeletal muscle. PMID- 15746180 TI - Identification of cardiac stem cells with FLK1, CD31, and VE-cadherin expression during embryonic stem cell differentiation. AB - We evaluated the expression of the FLK1, one of the lateral mesoderm early markers where cardiogenesis occurs, to characterize and isolate cardiac stem/progenitor cells from ES cells. Dissociated cells from embryoid bodies (EBs) on day 3, 4, or 5 were collected into two subpopulations with or without FLK1 expression and coculture on OP9 stromal cells was continued to examine whether contracting colonies came out or not. FLK1+ cells from EBs at days 3 and 4 formed spontaneous contracting colonies more efficiently than FLK1- cells on the same days, but not at day 5. Most contracting cardiac colonies derived from FLK1+cells mainly on day 4 were detected on endothelial cells along with hematopoietic cells. Further characterization of cells with these capabilities into three lineages revealed the FLK1+ CD31-VE-cadherin-phenotype. Our findings indicate that FLK1+cells, especially FLK1+ CD31-VE-cadherin-cells, could act as cardiohemangioblasts to form cardiac cells as well as endothelial cells and hematopoietic cells. PMID- 15746181 TI - Wound healing in rat cornea: the role of electric currents. AB - Human corneal epithelial cells respond rapidly following injury to restore the integrity of the ocular surface. What stimulates and guides cells to move into the wound to heal? One candidate is the wound-induced electric field. Using vibrating probe techniques, we provide detailed temporal and spatial mapping of endogenous electric currents at rat corneal wounds. We find Cl- and Na+ are the major components of electric currents in rat corneal wounds. Na+ is the major component of ionic transport in the resting (nonwounded) rat cornea and of the wound center leakage current, whereas Cl- is a more important component of the endogenous electrical current at the wound edges. Enhancing or decreasing Cl- flow with clinically approved pharmacological agents such as aminophylline, ascorbic acid, or furosemide increased or decreased endogenous wound electric currents, respectively. These changes in wound currents correlated directly with the rate of wound healing in vivo. Thus, pharmacologically enhancing or decreasing wound-induced electric currents increased and decreased wound healing rate, respectively. This may have wide-reaching and novel therapeutic potential in the management of wound healing and may help explain some mechanistic aspects of the effects of some clinically used agents. PMID- 15746182 TI - Oxidants in signal transduction: impact on DNA integrity and gene expression. AB - Physiological stimuli using reactive oxygen species (ROS) as second messengers caused nucleotide-specific base modifications in the hypoxic response element of the VEGF gene in lung vascular cells, with the 3' guanine of the HIF-1 DNA recognition sequence uniformly targeted. Modeling this effect by replacing the targeted guanine with an abasic site increased incorporation of HIF-1 and the bi functional DNA repair enzyme and transcriptional coactivator, Ref-1/Ape1, into the transcriptional complex and engendered more robust reporter gene expression. Oxidants generated in the context of physiological signaling thus affect nuclear DNA integrity and may facilitate gene expression by optimizing DNA-protein interactions. PMID- 15746183 TI - Src is necessary and sufficient for human airway smooth muscle cell proliferation and migration. AB - Airway smooth muscle (ASM) hypertrophy and hyperplasia, important pathological features in chronic severe asthma, likely contribute to irreversible airflow obstruction. Despite considerable research effort, the precise cellular mechanisms that modulate ASM growth remain unknown. Src, a nonreceptor tyrosine kinase proto-oncogene, reportedly modulates cell proliferative responses to growth factors, contractile agonists, and inflammatory mediators. Here, we show that Src activation is required for human ASM mitogenesis and motility. Platelet derived growth factor (PDGF), epidermal growth factor (EGF), and thrombin induce rapid activation of Src, and inhibition of Src induces a concentration-dependent abrogation of PDGF-, EGF-, and thrombin-induced ASM cell proliferation. Src immunoprecipitates had associated phosphatidylinositol 3-kinase, or PI3K, activation in response to PDGF and thrombin but not EGF. Further, Src activation is both necessary and sufficient for the stimulation of DNA synthesis as demonstrated by dominant negative Src inhibition of PDGF-, EGF-, and thrombin induced DNA synthesis. Human ASM cell migration was also attenuated by transfection of cells with dominant negative Src. Further, expression of constitutively active Src promoted cell migration. Collectively, these data demonstrate that Src modulates human ASM cell proliferation and migration, suggesting that Src may play an important role in promoting ASM cell growth and migration that occur in airway remodeling found in asthma and chronic obstructive pulmonary disease, or COPD. PMID- 15746184 TI - Proteomic analysis of neurofibrillary tangles in Alzheimer disease identifies GAPDH as a detergent-insoluble paired helical filament tau binding protein. AB - We performed proteomic analysis of neurofibrillary tangles (NFTs) obtained by laser capture microdissection from pyramidal neurons in hippocampal sector CA1 in patients with Alzheimer disease (AD) using liquid chromatography (LC)-mass spectrometry (MS)/MS. We discovered a total of 155 proteins in laser captured NFT's, 72 of which were identified by multiple unique peptides. Of these 72 proteins, 63 had previously unknown association with NFTs; one of these was glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We validated by immunohistochemistry that GAPDH co-localized with the majority of NFTs as well as plaque-like structures in AD brain and was co-immunoprecipitated by antibodies to abnormal forms of tau in AD, but not tau from AD temporal cortex. Characterization of GAPDH showed that it, along with phosphorylated tau and Abeta peptides, was present in detergent-insoluble fractions from AD temporal cortex but not from age-matched controls. These data are the first proteomic investigation of NFTs. Moreover, our results validate this approach by demonstrating that GAPDH, a glycolytic and microtubule binding protein, not only co-localized to NFTs and immunoprecipitated with PHF-tau, but also is one of the few proteins known to undergo conversion to a detergent-insoluble form in AD. PMID- 15746185 TI - The transport of cationic amino acids in human airway cells: expression of system y+L activity and transepithelial delivery of NOS inhibitors. AB - The transport of arginine has been characterized in human airway Calu-3 cells. As assessed with RT-PCR, Calu-3 cells express the genes for several transporters, such as the system y+-related SLC7A1, SLC7A2, and SLC7A4; the system y+L-related SLC7A6, SLC7A7, and SLC3A2; and the system B0,+-related SLC6A14. In polarized Calu-3 cell monolayers, apical arginine influx has a leucine-sensitive, sodium dependent component and a leucine- and lysine-resistant sodium-independent fraction. At the basolateral membrane, arginine transport was fully sodium independent and partially inhibited by leucine provided that sodium was present in the extracellular medium. Moreover, extracellular leucine trans-stimulated arginine efflux from the basolateral membrane in the presence, but not in the absence, of sodium. The transepithelial, apical to basolateral, arginine transport strictly depended on the presence of sodium and was markedly inhibited by apical leucine, but significantly trans-stimulated by the neutral amino acid added at the basolateral side. When added at the apical side, the NOS-inhibitors NMMA and NIL, CAA analogs with a free carboxyl group, markedly inhibited the apical arginine influx and the transepithelial flux of the cationic amino acid. The same compounds trans-stimulated basolateral arginine efflux. None of these effects were observed in the presence of the methyl ester analog NAME. The basolateral medium of Calu-3 cell monolayers, obtained after incubation in the presence of the three inhibitors at the apical side, inhibited the production of NO by activated murine macrophages. The inhibitory effect of the Calu-3 cell conditioned medium was time-dependent and markedly higher with NMMA and NIL than with NAME. Moreover, the NOS-inhibitory effect of the medium was significantly enhanced if NMMA and NIL, at the apical side, and basolateral leucine were simultaneously present during the conditioning procedure. These results indicate that 1) human airway epithelial cells express a functional system y+L at the basolateral membrane; 2) in this model, transepithelial arginine transport involves apical influx through system B0,+ and basolateral efflux through system y+L, and 3) the same transporters also perform an efficient transepithelial transport of amino acid-like NOS inhibitors. PMID- 15746186 TI - Inhibitory control of TGF-beta1 on the activation of Rap1, CD11b, and transendothelial migration of leukocytes. AB - Beta2-integrins are a family of dimeric adhesion molecules expressed on leukocytes. Their capacity to bind ligand is regulated by their state of activation. CD11b, an alphaMbeta2 integrin, is implicated in a number of physiological and pathological events such as inflammation, thrombosis, or atherosclerosis. The GTPase Rap1 is essential for its activation and could therefore play a strategic role in the regulation of leukocyte functioning. Because low levels of circulating TGF-beta have been linked with severe atherosclerosis, we have assessed the role of this cytokine in the regulation of Rap1 and CD11b activation in differentiated U937 cells and in human peripheral blood monocytes. TGF-beta1 caused a significant reduction in the expression of CD11b but not in the expression of other integrins tested. More importantly, TGF beta1 greatly reduced the capacity of PMA or chemokines to activate CD11b and Rap1, a phenomenon paralleled by a loss of the Epac transcript and a reduction in 8-pCPT-2'-O-Me-cAMP-mediated activation of Rap1. This inhibition diminished the capacity of monocytes to migrate across a monolayer of endothelial cells. The inhibitory effect of TGF-beta1 on Rap1 activity may exert a general protective influence against aberrant transendothelial migration, thereby holding inflammatory responses in check. PMID- 15746187 TI - Melatonin affects the temporal organization of the song of the zebra finch. AB - In birds and mammals, including humans, melatonin-binding sites are abundant in brain areas that have no known clock function. Although the role of such binding sites is still unclear, it is assumed that these sites link neural functions to circadian or circannual demands of neuroendocrine homeostasis and reproduction. To investigate a possible direct role of melatonin in motor control, we studied the song and neural song system of the zebra finch. Neurons of two sensory-motor areas of the descending song control circuit that are crucial for the organization of the song pattern, the HVC and RA, express the melatonin-1B receptor (Mel1B), while the hypoglossal motor neurons of the song circuit express melatonin-1C receptors (Mel1C). Application of melatonin to brain slices decreases the firing-rate of RA-neurons. Systemic administration of a Mel1B antagonist at the beginning of the night shortens the song and motif length and affects the song syllable lengths produced the next day. The temporal pattern of the song, however, does not undergo daily changes. Thus, melatonin is likely to affect a non-circadian motor pattern by local modulation of song control neurons and in consequence alters a sexual signal, the song of the zebra finch. PMID- 15746188 TI - Aldose reductase pathway mediates JAK-STAT signaling: a novel axis in myocardial ischemic injury. AB - The aldose reductase pathway has been demonstrated to be a key component of myocardial ischemia reperfusion injury. Previously, we demonstrated that increased lactate/pyruvate ratio, a measure of cytosolic NADH/NAD+, is an important change that drives the metabolic cascade mediating ischemic injury. This study investigated signaling mechanisms by which the aldose reductase pathway mediates myocardial ischemic injury. Specifically, the influence of the aldose reductase pathway flux on JAK-STAT signaling was examined in perfused hearts. Induction of global ischemia in rats resulted in JAK2 activation followed by STAT5 activation. Pharmacological inhibition of aldose reductase or sorbitol dehydrogenase blocked JAK2 and STAT5 activation and was associated with lower lactate/pyruvate ratio and lower protein kinase C activity. Niacin, known to lower cytosolic NADH/NAD+ ratio independent of the aldose reductase pathway inhibition, also blocked JAK2 and STAT5 activation. Inhibition of protein kinase C also blocked JAK2 and STAT5 activation. Transgenic mice overexpressing human aldose reductase exhibited increased JAK2 and STAT5 activation. Pharmacological inhibition of JAK2 reduced ischemic injury and improved functional recovery similar to that observed in aldose reductase pathway inhibited mice hearts. These data, for the first time, demonstrate JAK-STAT signaling by the aldose reductase pathway in ischemic hearts and is, in part, due to changes in cytosolic redox state. PMID- 15746189 TI - The antiangiogenic factor, 16-kDa human prolactin, induces endothelial cell cycle arrest by acting at both the G0-G1 and the G2-M phases. AB - The 16-kDa N-terminal fragment of human prolactin (16K hPRL) is a potent antiangiogenic factor that has been shown to prevent tumor growth in a xenograph mouse model. In this paper we first demonstrate that 16K hPRL inhibits serum induced DNA synthesis in adult bovine aortic endothelial cells. This inhibition is associated with cell cycle arrest at both the G(0)-G(1) and the G(2)-M phase. Western blot analysis revealed that 16K hPRL strongly decreases levels of cyclin D1 and cyclin B1, but not cyclin E. The effect on cyclin D1 is at least partially transcriptional, because treatment with 16K hPRL both reduces the cyclin D1 mRNA level and down-regulates cyclin D1 promoter activity. This regulation may be due to inhibition of the MAPK pathway, but it is independent of the glycogen synthase kinase-3beta pathway. Lastly, 16K hPRL induces the expression of negative cell cycle regulators, the cyclin-dependent kinase inhibitors p21(cip1) and p27(kip1). In summary, 16K hPRL inhibits serum-induced proliferation of endothelial cells through combined effects on positive and negative regulators of cell cycle progression. PMID- 15746190 TI - Increased expression of the glucocorticoid receptor-A translational isoform as a result of the ER22/23EK polymorphism. AB - One of the most intriguing polymorphisms in the GR [glucocorticoid (GC) receptor] gene is in codons 22 and 23 [GAGAGG(GluArg) --> GAAAAG (GluLys)]. This polymorphism is associated with a reduced GC sensitivity, a better metabolic and cardiovascular health profile, and an increased survival rate. Recently, Yudt and Cidlowski reported that two different methionine codons in the GR mRNA may be used as initiation codon: AUG-1 and AUG-27, resulting in two isoforms, the GR-A and the GR-B proteins, respectively. They also showed that the GR-B protein had a stronger transactivating effect in transient transfection experiments. In this study, we elucidated the molecular basis for the reduced GC sensitivity by investigating the influence of the ER22/23EK polymorphism on synthesis of GR-A and GR-B by expressing them independently from constructs with and without the polymorphic site. Binding studies with [(3)H]-dexamethasone and transactivation studies showed that, when the ER22/23EK polymorphism is present, approximately 15% more GR-A protein was expressed, whereas total GR levels (GR-A + GR-B) were not affected. These results show that the transcriptional activity in GR(ER22/23EK) carriers is decreased because more of the less transcriptionally active GR-A isoform is formed. This is probably caused by altered secondary mRNA structure. PMID- 15746192 TI - Defective calmodulin-mediated nuclear transport of the sex-determining region of the Y chromosome (SRY) in XY sex reversal. AB - The sex-determining region of the Y chromosome (SRY) plays a key role in human sex determination, as mutations in SRY can cause XY sex reversal. Although some SRY missense mutations affect DNA binding and bending activities, it is unclear how others contribute to disease. The high mobility group domain of SRY has two nuclear localization signals (NLS). Sex-reversing mutations in the NLSs affect nuclear import in some patients, associated with defective importin-beta binding to the C-terminal NLS (c-NLS), whereas in others, importin-beta recognition is normal, suggesting the existence of an importin-beta-independent nuclear import pathway. The SRY N-terminal NLS (n-NLS) binds calmodulin (CaM) in vitro, and here we show that this protein interaction is reduced in vivo by calmidazolium, a CaM antagonist. In calmidazolium-treated cells, the dramatic reduction in nuclear entry of SRY and an SRY-c-NLS mutant was not observed for two SRY-n-NLS mutants. Fluorescence spectroscopy studies reveal an unusual conformation of SRY.CaM complexes formed by the two n-NLS mutants. Thus, CaM may be involved directly in SRY nuclear import during gonadal development, and disruption of SRY.CaM recognition could underlie XY sex reversal. Given that the CaM-binding region of SRY is well-conserved among high mobility group box proteins, CaM-dependent nuclear import may underlie additional disease states. PMID- 15746191 TI - Prolactin and estrogen enhance the activity of activating protein 1 in breast cancer cells: role of extracellularly regulated kinase 1/2-mediated signals to c fos. AB - Despite the important roles of both prolactin (PRL) and 17beta-estradiol (E2) in normal mammary development as well as in breast cancer, and coexpression of the estrogen receptor (ER) and PRL receptor in many mammary tumors, the interactions between PRL and E2 in breast cancer have not been well studied. The activating protein 1 (AP-1) transcription factor, a known regulator of processes essential for normal growth and development as well as carcinogenesis, is a potential site for cross-talk between these hormones in breast cancer cells. Here we demonstrate that PRL and E2 cooperatively enhance the activity of AP-1 in MCF-7-derived cells. In addition to the acute PRL-induced ERK1/2 activation, PRL and E2 also individually elicited delayed, sustained rises in levels of phosphorylated p38 and especially ERK1/2. Together, these hormones increased the dynamic phosphorylation of ERK1/2 and c-Fos, and induced c-fos promoter activity. Synergistic activation of the transcription factor, Elk-1, reflected the PRL-E2 interaction at ERK1/2 and is a likely mechanism for activation of the c-fos promoter via the serum response element. The enhanced AP-1 activity resulting from the interaction of these hormones may increase expression of many target genes that are critical for oncogenesis and may contribute to neoplastic progression. PMID- 15746193 TI - Release of methyl CpG binding proteins and histone deacetylase 1 from the Estrogen receptor alpha (ER) promoter upon reactivation in ER-negative human breast cancer cells. AB - Estrogen receptor alpha (ER) is an epigenetically regulated gene. Inhibitors of DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) synergistically activate the methylated ER gene promoter in ER-negative MDA-MB-231 human breast cancer cells. Chromatin immunoprecipitation was used to examine the chromatin status and repressor complex associated with silenced ER and changes in the key regulatory factors during reactivation by inhibitors of DNMT (5-aza-2' deoxycytidine) and HDAC (trichostatin A). The silencing of ER due to CpG hypermethylation correlates with binding of specific methyl-binding proteins, DNMTs, and HDAC proteins. Inhibition of HDAC activity by trichostatin A results in the accumulation of hyperacetylated core histones. The activation of ER gene expression by 5-aza-2'-deoxycytidine also involves the release of the repressor complex involving various methyl-binding proteins, DNMTs, and HDAC1. HDAC and DNMT inhibitors modulate histone methylation at H3-K9 and H3-K4 to form a more open chromatin structure necessary for reactivation of silenced ER transcription. Together these results impart a better understanding of molecular mechanisms of chromatin remodeling during ER reactivation by DNMT and HDAC inhibitors. These findings will aid in the application of agents targeting epigenetic changes in the treatment of breast cancer. PMID- 15746194 TI - The vascular endothelial growth factor (VEGF) +405G>C 5'-untranslated region polymorphism and increased risk of endometriosis in South Indian women: a case control study. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis and vascular permeability, is known to play a key role in the pathophysiology of endometriosis. METHODS AND RESULTS: The single nucleotide polymorphisms, -460C>T and +405G>C, in the 5'-untranslated region of the VEGF gene were tested for association in a case-control study of 215 affected women and 210 women with no evidence of disease. All the women were of South Indian origin and ascertained from the same infertility clinic. The genotype and allele frequencies of the -460C>T polymorphism did not differ significantly between cases and controls. In contrast, the genotype (P = 0.002) and allele (P = 0.001) frequencies of the +405G>C polymorphism showed a significant difference between cases and controls. The +405 GG genotype was found more often in patients with an endometrioma >3 cm compared to controls. The frequency of the -460T/+405C haplotype (P = 0.016) was significantly lower in affected women compared to controls. CONCLUSIONS: The -460T/+405C haplotype in the VEGF gene, which is associated with lower promoter activity, was significantly less common in women with endometriosis than in controls. These data suggest that the +405G allele may influence the likelihood of a woman developing the disease. PMID- 15746195 TI - Ovarian response to standard gonadotrophin stimulation for IVF is decreased not only in older but also in younger women in couples with idiopathic and male subfertility. AB - BACKGROUND: With the occasional reports of unexpectedly poor ovarian response to controlled ovarian hyperstimulation (COH) for IVF in young normally cyclic women in mind, we studied age-related ovarian response to COH in a group of women who underwent standard IVF. METHODS: Ovarian response to COH was defined as the number of follicles > or = 14 mm on the day of hCG administration. Ovarian response to COH was analysed by multiple regression analysis with woman's age and basal FSH concentration as explanatory variables in a prospective cohort of patients with idiopathic and mild male factor subfertility (n = 85), and additionally in a large retrospective cohort of women with unexplained, mild male and tubal subfertility (n = 1155), with age as explanatory variable. RESULTS: Ovarian response to COH was associated significantly with age (P < 0.001) and basal FSH concentration (P = 0.002). However, in women with idiopathic or mild male subfertility, in both cohorts the relationship took the form of an inverted U-shape with both older and--surprisingly--young women having a reduced ovarian response (P < 0.001). Maximum ovarian response was around the age of 28 years. In women with tubal infertility, there was only a linear decline of ovarian response with age. CONCLUSION: It is hypothesized that diminished ovarian response to COH in IVF is the very first sign of ovarian ageing in young women diagnosed with idiopathic and mild male subfertility. PMID- 15746196 TI - Comparison of the survival of human biopsied embryos after cryopreservation with four different methods using non-transferable embryos. AB - BACKGROUND: The standard embryo cryopreservation method is still less than optimal for biopsied embryos. The aim of this study was to compare the survival of biopsied embryos cryopreserved with four different methods using non transferable embryos. METHODS: Abnormal embryos from one or three pronuclei and spare embryos of grade 3 and 4 were used for this study. Non-biopsied embryos were cryopreserved using the standard method as control. Biopsied embryos were cryopreserved using four methods as follows: standard method, modified freezing method, modified thawing method and vitrification. Blastomere survival and blastulation of frozen-thawed embryos were compared between the different methods. RESULTS: The proportion of embryos with > or = 50% blastomere survival and total blastomere survival rate of biopsied embryos were significantly higher with vitrification than the other three methods. Both the modified freezing and modified thawing methods had significantly higher embryo survival and total blastomere survival rates than standard methods. However, there was no significant difference in blastulation of surviving embryos in all the five groups. CONCLUSIONS: Non-transferable embryos derived from clinical IVF/ICSI are useful for evaluation of the optimal freezing procedures for biopsied embryos. Vitrification increases the survival rate of human biopsied embryos above standard and modified cryopreservation methods. PMID- 15746197 TI - Search for mutations involved in human globozoospermia. AB - BACKGROUND: Globozoospermia is a severe form of teratozoospermia characterized by round-headed sperm with an absence of acrosomes. Family cases of globozoopermia suggest that this pathology has genetic origins, but the mode of inheritance remains unknown. So far, no responsible genes have been identified. Recently, a mouse lacking the casein kinase IIalpha' (encoded by the Csnk2a2 gene) was described. This mutant mouse presents a single phenotype reminiscent of that seen in human globozoospermia. Interestingly, the fission yeast orthologue (orb5) exhibits, when mutated, a spherical phenotype. Casein kinase II is a heterotetramer, composed of two catalytic subunits alpha or alpha' and two regulatory beta subunits (encoded by the Csnk2b gene). METHODS AND RESULTS: Based on the evolution conservation, phenotypes observed in mouse and yeast mutant and the structure of casein kinase II, we analysed Csnk2a2 and Csnk2b genes in six patients with globozoospermia and 10 fertile controls. Genomic DNA was extracted from peripheral blood and PCR was performed to amplify Csnk2a2 and Csnk2b genes before sequencing. CONCLUSION: No mutation was identified among these six patients. Further work is needed, with a larger patient data set, to identify putative genes involved in this form of male infertility. PMID- 15746198 TI - Interleukin 1alpha and tissue-lytic matrix metalloproteinase-1 are elevated in ectopic endometrium of patients with endometriosis. AB - BACKGROUND: Matrix metalloproteinases (MMP) play an essential role in tissue remodelling and menstruation and appear to be regulated by cytokines such as interleukin-1alpha (IL-1alpha). In order to investigate their role in the pathogenesis of endometriosis, the aim of the present study was to compare the protein localization of matrix metalloproteinase-1 (MMP-1) and of its main stimulatory cytokine IL-1alpha in eutopic and dystopic endometrium of patients with endometriosis. METHODS: MMP-1 and IL-1alpha protein localization was analysed retrospectively in paired paraffin-embedded tissue biopsies obtained simultaneously from the endometrial cavity and from endometrial lesions of 37 patients with peritoneal or ovarian endometriosis and in cycling endometria from 37 women without endometriosis. Protein localization was demonstrated by immunohistochemistry; antibody specificity was confirmed by western blot analysis. RESULTS: MMP-1 and IL-1alpha protein staining in women suffering from endometriosis was significantly more pronounced in endometriotic lesions than in eutopic endometrium. This held true for both epithelial MMP-1 and IL-1alpha staining (P < 0.006 and P < 0.001), and for stromal MMP-1 and IL-1alpha staining (P < 0.001 and P < 0.001). Furthermore, stromal MMP-1 and IL-1alpha were significantly co-expressed in dystopic endometriotic tissue (P = 0.045). Endometrial MMP-1 and IL-1alpha protein expression pattern in eutopic endometrium from women suffering from endometriosis, however, did not differ significantly from the pattern seen in healthy women. CONCLUSIONS: The increased expression of both matrix-degrading MMP-1 and its major stimulatory cytokine IL-1alpha in endometriotic lesions and the selective co-expression in the stroma of endometriotic foci clearly suggests their involvement in the pathogenic mechanisms leading to local invasion and tissue destruction. PMID- 15746199 TI - Concentrations of leptin and C-reactive protein in serum and follicular fluid during assisted reproductive cycles. AB - BACKGROUND: There are only a few studies that have investigated inflammatory processes during ovarian hyperstimulation, with contradictory results especially concerning outcome. The aim of the study was to investigate the inflammatory markers C-reactive protein and leptin in serum and follicular fluid and to correlate these with the outcome. METHODS: One hundred and sixty-two gonadotrophin stimulated cycles were evaluated. Serum concentrations of leptin and C-reactive protein were measured at the initiation of stimulation, on the day of hCG administration or the day before, and on the day of oocyte retrieval. They were also determined in the follicular fluid. RESULTS: Serum leptin and C reactive protein levels increased significantly during stimulation until the day of oocyte pick up, but following different patterns. After stimulation, they correlated with each other in serum and follicular fluid, but not with estradiol or progesterone concentration, embryo quality, or the pregnancy rate. CONCLUSIONS: Leptin and C-reactive protein levels change significantly during assisted reproductive treatment. In contrast to estradiol they are, however, not a marker of success. PMID- 15746200 TI - Influence of early ICSI-derived embryo sHLA-G expression on pregnancy and implantation rates: a prospective study. AB - BACKGROUND: We have previously reported the retrospective observation that when at least one embryo, transferred on day 3, expressed sHLA-G above the geometric mean (sHLA-G+) 46 h post-ICSI, there was a marked improvement in both pregnancy (PR) and implantation (IR) rates. METHODS: The media surrounding individual embryos derived from ICSI performed on oocytes from 482 women < or =43 years of age were tested for sHLA-G expression by specific ELISA. RESULTS: We report here prospective results showing improved IVF results following the transfer of 'good quality' embryos (7-9 cells with <20% fragmentation) by preferentially including at least one sHLA-G+ embryos. PR and IR for women < or =38 years were 63% and 32% when one transferred embryo was sHLA-G+, and 69% and 36% when at least two embryos were sHLA-G+. When none of the embryos transferred was sHLA-G+, PR and IR were 25% and 13%, respectively. Comparable PR and IR for women 39-43 years were 29% and 11% when none of the transferred embryos were sHLA-G+; 38% and 15% when at least one sHLA-G+ embryo was transferred; and 61% and 26% when at least two 2 sHLA-G+ embryos were transferred. The data were stratified by patient age. CONCLUSIONS: PR and IR increased with the addition of each sHLA-G+ embryo, regardless of age. While there are significant barriers to routine embryo sHLA-G testing, we believe that if implemented, this would provide a mechanism for optimizing IVF PR while minimizing the risk of multiple pregnancies. PMID- 15746201 TI - The presence of functional mannose receptor on macrophages at the maternal-fetal interface. AB - BACKGROUND: The mannose receptor (MR) is involved in the initiation of the immune response and regulation of homeostasis during inflammation and tissue remodeling. METHODS: Distribution, endocytosis and possible natural ligand tumor associated glycoprotein-72 (TAG-72) for the MR have been examined by immunohistology, immunocytochemistry and flow cytometry at the maternal-fetal interface, characterized by extensive tissue remodeling. RESULTS: Contrary to disseminated distribution of the MR positive (MR+) cells in term placenta, the MR+ cells of early pregnancy decidua intimately surrounded glands and followed tissue distribution of CD14 positive cells. The mannose receptor was present on freshly isolated first trimester decidual mononuclear cells and distributed mostly on macrophages (77.08 +/- 10.55%, mean +/- SD). The expression of the MR on CD14 positive cells decreased following 18 h culture (P < 0.01) and was accompanied by the reduction of fluorescein isothiocyanate (FITC)-dextran uptake. PAM-1 anti-MR antibody, mannan and TAG-72 reduced FITC-dextran uptake by decidual macrophages. CONCLUSIONS: These data indicate that the MR+ macrophages, surrounding early decidual glands, are able to internalize ligands for carbohydrate recognition domain of the receptor, including decidual secretory phase mucin TAG-72. PMID- 15746202 TI - Are early life factors responsible for international differences in adult blood pressure? An ecological study. AB - BACKGROUND: Although low birthweight has been related to higher mean blood pressure in individuals, there have been very few studies on the contribution of the 'fetal origins hypothesis' to international variations in mean blood pressure. METHODS: Standardized data on mean blood pressure levels in adults aged 20-29 years (available from the INTERSALT study countries) were related to data on mean birthweight and infant mortality rates (IMRs) approximately 25 years earlier (available from the World Health Organization and the United Nations statistics for 24 of these countries). RESULTS: Population mean systolic pressure was inversely correlated with IMR (r = -0.66, P < 0.001) and positively correlated with mean birthweight (r = 0.36, P = 0.108). These correlations were little affected by adjustment either for current body mass index or for current sodium, potassium, and alcohol intake. CONCLUSIONS: The directions of these associations suggest that low mean birthweight and high infant mortality are not important determinants of high population mean adult blood pressure levels. PMID- 15746203 TI - Commentary: how does socioeconomic disadvantage during childhood damage health in adulthood? Testing psychosocial pathways. PMID- 15746205 TI - Odol, Autobahne and a non-smoking Fuhrer: reflections on the innocence of public health. PMID- 15746204 TI - Explaining socioeconomic inequality in mortality among South Koreans: an examination of multiple pathways in a nationally representative longitudinal study. AB - BACKGROUND: South Korea has a different cause-specific structure of mortality compared with North America and northern European countries where studies on pathways to socioeconomic mortality inequalities have been performed. We examined the ability of multiple pathways to explain socioeconomic differentials in all cause mortality in South Korea. METHODS: The 1998 National Health and Nutrition Survey data of South Korea were linked to data on mortality. The socioeconomic position (SEP) indicator was household income. Twelve variables represented biological risk factors (body mass index, systolic blood pressure, cholesterol, and glucose), health behaviours (smoking, alcohol consumption, and regular exercise), psychosocial factors (feelings of sadness and depression, perceived level of stress, and marital status), and early life exposures (education and adulthood height). RESULTS: Mortality differentials by income level did not decrease after exclusion of subjects with severe chronic illness or functional limitation. Biological risk factors, health behaviours, and psychosocial factors caused minor reductions in relative risk for income levels. The ability of early life exposures to explain socioeconomic differentials in mortality was greater than that of biological risk factors, health behaviours, and psychosocial factors. CONCLUSIONS: The contribution of multiple pathways to socioeconomic differentials in all-cause mortality may vary in place with the different cause specific structure of mortality. Future studies with specific pathway variables and specific disease outcomes would provide better understanding of causal mechanisms between SEP and health. PMID- 15746206 TI - Growth in early life and childhood IQ at age 11 years: the Newcastle Thousand Families Study. AB - BACKGROUND: It has been suggested that in addition to genetic factors, fetal and post-natal growth influence childhood cognition, although it is unclear whether such an effect continues throughout childhood. This study aimed at investigating the potential relationships between childhood IQ at age 11 years and birth weight and height at the ages of 9 and 13 years, after adjusting for the confounding factors available to this investigation. METHODS: The Newcastle Thousand Families study, a prospectively followed cohort, originally consisted of all 1142 births in the city of Newcastle in May and June 1947. Using data on 733 members of this cohort, we investigated the associations between IQ at age 11, and birth weight and height at ages 9 and 13 years. RESULTS: Birth weight showed no association with childhood IQ. However, height at age 9 years was a significant predictor of childhood IQ after adjusting for socioeconomic status (standardized regression coefficient b = 2.6, 95% CI 1.6-3.6, P < 0.0001). Height at age 13 was also a significant predictor of IQ after adjusting for socioeconomic status (b = 3.4, 95% CI 2.3-4.4, P = 0.001), and explained an additional 2.5% of the variation in IQ scores to that already explained by socioeconomic status and height at age nine. CONCLUSIONS: These results suggest a continuing effect of post-natal growth on childhood cognition beyond the age of 9 years. Post-natal growth, which may be influenced by genetic factors and nutrition and socioeconomic circumstances in childhood, may be more important than fetal growth in terms of childhood cognition. PMID- 15746207 TI - Commentary: understanding religious involvement and mortality risk in the United States: comment on Bagiella, Hong, and Sloan. PMID- 15746209 TI - Social capital and the history of mortality in Britain. PMID- 15746210 TI - Commentary: maternal diet during pregnancy and blood pressure in the offspring. PMID- 15746211 TI - Repeated exposure to water avoidance stress in rats: a new model for sustained visceral hyperalgesia. AB - Chronic stress plays an important role in the development and exacerbation of symptoms in functional gastrointestinal disorders. To better understand the mechanisms underlying this relationship, we aimed to characterize changes in visceral and somatic nociception, colonic motility, anxiety-related behavior, and mucosal immune activation in rats exposed to 10 days of chronic psychological stress. Male Wistar rats were submitted daily to either 1-h water avoidance (WA) stress or sham WA for 10 consecutive days. The visceromotor response to colorectal distension, thermal somatic nociception, and behavioral responses to an open field test were measured at baseline and after chronic WA. Fecal pellets were counted after each WA stress or sham WA session as a measure of stress induced colonic motility. Colonic samples were collected from both groups and evaluated for structural changes and neutrophil infiltration, mast cell number by immunohistochemistry, and cytokine expression by quantitative RT-PCR. Rats exposed to chronic WA (but not sham stress) developed persistent visceral hyperalgesia, whereas only transient changes in somatic nociception were observed. Chronically stressed rats also exhibited anxiety-like behaviors, enhanced fecal pellet excretion, and small but significant increases in the mast cell numbers and the expression of IL-1beta and IFN-gamma. Visceral hyperalgesia following chronic stress persisted for at least a month. Chronic psychological stress in rats results in a robust and long-lasting alteration of visceral, but not somatic nociception. Visceral hyperalgesia is associated with other behavioral manifestations of stress sensitization but was only associated with minor colonic immune activation arguing against a primary role of mucosal immune activation in the maintenance of this phenomenon. PMID- 15746212 TI - Phosphatidylinositol 3-kinase-dependent signaling modulates taurochenodeoxycholic acid-induced liver injury and cholestasis in perfused rat livers. AB - Taurochenodeoxycholic acid (TCDCA), but not glycochenodeoxycholic acid (GCDCA), activates a phosphatidylinositol 3-kinase (PI3-K)-mediated survival pathway in vitro. Here, the effects of PI3-K inhibition on TCDCA- and GCDCA-induced hepatocellular injury, apoptosis, and bile secretion were examined in the intact liver. In isolated perfused rat livers, bile flow was determined gravimetrically. Hepatovenous lactate dehydrogenase and alanine aminotransferase efflux as markers of liver integrity and biliary secretion of 2,4-dinitrophenyl-S-glutathione (DNP GS) were determined photometrically. Apoptosis was assessed by immunohistochemistry of active caspase-3 and cytokeratin 18 in liver tissue. Phosphorylation of protein kinase B (PKB/Akt) as a readout of PI3-K activity was determined by immunoblot analysis. Bile acid concentrations were determined by gas chromatography. TCDCA (25 muM) induced moderate liver injury by hepatocellular apoptosis and distinctly reduced bile flow and DNP-GS secretion. In contrast, GCDCA (25 muM) induced severe liver injury by extensive hepatocyte apoptosis. TCDCA strongly activated PI3-K, whereas GCDCA did not markedly affect PI3-K activity. Inhibition of PI3-K by 100 nM wortmannin enhanced TCDCA-induced liver injury and apoptosis and tended to aggravate the cholestatic effect of TCDCA. In contrast, wortmannin reduced GCDCA-induced liver injury and apoptosis. Bile acid uptake tended to be reduced by wortmannin. The cholestatic effect of GCDCA was aggravated by wortmannin. Inhibition of PI3-K markedly aggravated TCDCA induced but not GCDCA-induced liver damage and hepatocyte apoptosis. Thus TCDCA appears to block its inherent toxicity by a PI3-K-dependent survival pathway in the intact liver. PMID- 15746213 TI - Extracellular cysteine/cystine redox regulates the p44/p42 MAPK pathway by metalloproteinase-dependent epidermal growth factor receptor signaling. AB - Previous research shows that stimulation of proliferation of colon carcinoma (Caco-2) cells by a more reduced extracellular cysteine/cystine (Cys/CySS) redox state occurs with no apparent effect on intracellular glutathione and that this stimulation is lost on addition of epidermal growth factor. The purpose of the present study was to determine whether a more reduced extracellular Cys/CySS redox state activates the mitogenic p44/p42 mitogen-activated protein kinase (MAPK) pathway and whether this is signaled through the epidermal growth factor receptor (EGFR). Caco-2 cells were exposed to a range of physiological extracellular redox conditions from -150 to 0 mV. In the absence of added growth factors, the most reduced (-150 mV) redox state induced an 80% increase in EGFR phosphorylation, and this was followed by a marked increase in phosphorylation of p44/p42 MAPK. Inhibitors of EGFR (AG1478) and p44/p42 MAPK (U0126) phosphorylation blocked redox-dependent p44/p42 phosphorylation, indicating that signaling occurred by EGFR. These effects were inhibited by pretreatment with a nonpermeant alkylating agent, showing that signaling involved thiols accessible to the extracellular space. The EGFR ligand TGF-alpha was increased in culture medium at more reduced redox states. Redox-dependent phosphorylation of EGFR was completely prevented by a metalloproteinase inhibitor (GM6001), and an antibody to TGF-alpha partially inhibited the phosphorylation of p44/p42 MAPK by redox. Thus the data show that a redox-dependent activation of metalloproteinase can stimulate the mitogenic p44/p42 MAPK pathway by a TGF-alpha-dependent mechanism. Because Cys availability and Cys/CySS redox are dependent on nutrition, disease, and environmental exposures, the results suggest that cell proliferation could be influenced physiologically by Cys-dependent redox effects on growth factor signaling pathways. PMID- 15746214 TI - Characterization of 5-HT7-receptor-mediated gastric relaxation in conscious dogs. AB - We aimed to evaluate the gastric relaxant capacity of the 5-HT(1/7)-receptor agonist 5-carboxamidotryptamine (5-CT) in conscious dogs and to clarify the mechanism of action by use of selective antagonists, vagotomy, and in vitro experiments. A barostat enabled us to monitor the intragastric volume in response to different treatments (intravenously administered) before and after supradiaphragmatic vagotomy [results presented as the maximum volume change after treatment (mean; n = 5-11)]. In vitro experiments were performed with isolated muscle strips cut from four different stomach regions of the vagotomized dogs [results were fitted to the operational model of agonism to determine the efficacy parameter tau (n = 5)]. 5-CT (0.5-10 microg/kg) caused a dose-dependent gastric relaxation (29-267 ml) that was completely blocked by the selective 5 HT(7)-receptor antagonist SB-269970 (50 microg/kg). After vagotomy, the relaxation to 10 microg/kg 5-CT was significantly less pronounced (73 vs. 267 ml; P < 0.05) but still blocked by SB-269970, whereas the response to the nitric oxide donor nitroprusside was similar to that before vagotomy (178 vs. 218 ml). In vitro, 5-CT concentration dependently inhibited the PGF(2alpha)-contracted muscle strips before and after vagotomy. Although before and after vagotomy the response in every region was mediated by 5-HT(7) receptors (apparent affinity dissociation constant: SB-269970, 8.2-8.6 vs. 8.3-8.6, respectively), the response after vagotomy was less efficacious (log tau: 1.9 to 0.5 vs. 1.4 to 0.1). The results indicate that the 5-CT-induced proximal stomach relaxation in conscious dogs before and after vagotomy is mediated via 5-HT(7) receptors. The decreased efficacy of 5-CT in vitro after vagotomy is probably related to vagotomy-induced changes in receptor density or coupling efficiency and provides a possible explanation for the decreased in vivo response to 5-CT after vagotomy. PMID- 15746215 TI - Prevention and promotion in decentralized rural health systems: a comparative study from northeast Brazil. AB - Policies to reform health care provision often combine the organizational restructuring of decentralization with ideological restructuring through a new model of health care that gives greater weight to prevention and promotion. Decentralization provides a discretionary space to the local health system to define and develop its own activities. The central policy aim to shift the model of health care therefore must rely on incentives rather than directives and is likely to result in variation at local levels in the extent and mode of its implementation. The local processes affecting variation in local implementation of policies for prevention and promotion have not been studied in a developing country. This study does so by comparing two rural health systems with different levels of prevention and promotion activities in one of the poorest regions of Brazil, Ceara State in the northeast. The health system with greater activities of prevention and promotion also has a more advanced stage of decentralization, but this is in combination with many other, interacting influences that differentiate the two health systems' ability to adopt and implement new approaches. While beyond the scope of this paper to detail options for regional and national managers to encourage the adoption of a greater focus on prevention and promotion, it is clear that strategies needs to target not only the vision and actions of local health system staff, but critically also the expectations of the local population and the attitudes of local government. PMID- 15746216 TI - Changes in access to health care in China, 1989-1997. AB - The post-1979 period in China has seen the implementation of reforms that dismantled much of the Maoist era social welfare system and permitted a significant reallocation of society's resources. The result has been rapid but uneven economic development that has profoundly altered the environment within which consumers make health investment decisions. Many studies report significant and apparently non-random reductions in health care utilization during this period. Scholars have tended to focus on the loss of insurance coverage and the growth of fees for services in explaining such reductions. An alternative explanation is growing inequality in access to care. This possibility has not received much research attention. As a result, our understanding of the patterns of changes in health care access, and of the types of populations that have been most adversely affected, has been rather limited. This research examines the distribution of the changes in several indicators of access to health care across communities during the period 1989 to 1997. We find evidence of relatively uneven changes to these indicators. Money charges for routine services increased consistently, though this trend was less pronounced in lower-income communities. Most communities experienced reductions in travel distance to clinics but increases in distance to hospitals. There were major improvements to the quality of care in wealthier rural areas, but not in poorer villages. Wealthier villages experienced less improvement in waiting time and drug availability. These trends appear to be closely associated with changing economic circumstances during the reform era. PMID- 15746217 TI - Penalizing patients and rewarding providers: user charges and health care utilization in Vietnam. AB - The introduction of a comprehensive system of user charges in 1995 provided public health facilities in Vietnam, especially hospitals, with a growing source of revenue. By 1998 revenues from user charges accounted for 30% of public hospital revenues. Increasingly, provider incomes have relied on fee revenues and provision-based bonuses, the effect of which is that a poorly regulated fee-for service system has replaced a salary system based upon a centrally determined global budget. This paper examines the potential influence of providers' on the use of publicly provided health services. Using facility-based data over the period 1996-98, the relative contribution of treatment intensity is compared and contrasted under the two sources of hospital revenues from patients, namely a user charge system and a third party payment system based on fee-for-services. The primary focus of the comparison is on the treatment intensity for all hospital contacts, hospital admissions and the length of hospital stays, decisions normally taken by the providers and over which patients have little or no influence. The results indicate that growth in patient revenues was associated with large increases in intensity. The growth in intensity was more pronounced in the case of inpatient contacts. Moreover, both the admission rate and the length of hospital stay were far higher for better off individuals than for the poor, and greater for the insured than the uninsured. The increase in the intensity of hospital care for both health insurance enrollees and the uninsured can be seen as, among other things, an attempt on the part of providers to increase revenue from health insurance premiums and user charges in the face of a shrinking share of public resources allocated to hospitals, and low wages and salaries. PMID- 15746218 TI - Abolition of cost-sharing is pro-poor: evidence from Uganda. AB - OBJECTIVE: To document the effects of the abolition of user fees on utilization of health services in Uganda with emphasis on poor and vulnerable groups. METHODS: A longitudinal study using quantitative and qualitative methods was carried out in 106 health facilities across the country. Health records were reviewed to determine trends in overall utilization patterns and use among vulnerable groups. A modification of wealth ranking as defined by the Uganda Poverty Participatory Assessment Project was used to categorize households by socio-economic status in order to compare utilization by the poor against that of other socio-economic groups. FINDINGS: There was a marked increase in utilization in all population groups that was fluctuating in nature. The increase in utilization varied from 26% in public referral facilities in 2001, rising to 55% in 2002 compared with 2000. The corresponding figures for the lower level facilities were 44% and 77%, respectively. Increase in utilization among the poor was more than for other socio-economic categories. Women utilized health services more than men both before and after cost-sharing. Higher increases in utilization were noted among the over-five age group compared with the under-fives. There were no increases in utilization for preventive and inpatient services. With respect to quality of care, there were fewer drug stock-outs in 2002 compared with 2000 and 2001. There was no deterioration of other indicators such as cleanliness, compound maintenance and staff availability reported. CONCLUSION: The study suggests that there is a financial barrier created by cost-sharing that decreases access to services, especially among the poor in Uganda. However, further studies are needed to clarify issues of utilization by age and gender. PMID- 15746219 TI - Evaluation of a home-based voluntary counselling and testing intervention in rural Uganda. AB - BACKGROUND: Uptake of HIV test results from an annual serosurvey of a population study cohort in rural southwestern Uganda had never exceeded 10% in any given year since inception in 1989. An intervention offering counselling and HIV results at home was conducted in four study villages following the 2001 serosurvey round, and followed by a qualitative evaluation exploring nature of demand and barriers to knowing HIV status. METHODS: Data from annual serosurveys and counsellor records are analyzed to estimate the impact of the intervention on uptake of HIV test results. Textual data are analyzed from 21 focus group discussions among counsellors, and men and women who had received HIV test results, requested but not yet received, and never requested; and 34 in-depth interviews equally divided among those who had received test results either from counselling offices and homes. RESULTS: Offering HIV results at home significantly increased uptake of results from 10 to 37% for all adults aged 15 (p<0.001), and 46% of those age 25 to 54. Previous male advantage in uptake of test results was effectively eliminated. Focus group discussions and in-depth interviews highlight substantial non-monetary costs of getting HIV results from high-visibility public facilities prior to intervention. Inconvenience, fear of stigmatization, and emotional vulnerability of receiving results from public facilities were the most common explanations for the relative popularity of home based voluntary counselling and testing (VCT). It is seen as less appropriate for youth and couples with conflicting attitudes toward testing. CONCLUSIONS: Home delivery of results revealed significantly higher demand to know HIV status than stubbornly low uptake figures from the past would suggest. Integrating VCT into other services, locating testing centres in less visible surroundings, or directly confronting stigma surrounding testing may be less expensive ways to reproduce increased uptake with home VCT. PMID- 15746220 TI - Increasing contraceptive use in rural Pakistan: an evaluation of the Lady Health Worker Programme. AB - Past efforts to promote family planning in Pakistan have been disappointing, but between 1990-91 and 2000-01 contraceptive use has more than doubled. This rise has coincided with a concerted effort on the part of the Pakistani government to increase access to contraceptive services, particularly in rural areas. The Lady Health Worker Programme (LHWP), initiated under the Ministry of Health in the early 1990s, aimed at integrating family planning into the doorstep provision of primary health care. This paper presents findings from the first national evaluation of this Programme. Data are analyzed from a random sample survey of 4277 women living in households served by the LHWP and those living in control areas. Logistic regression analysis was performed to determine the effect of the Programme on the uptake of modern reversible contraceptive methods, controlling for other independent variables. The data provide strong evidence that the LHWP has succeeded in increasing modern contraceptive use among rural women. Women served by Lady Health Workers are significantly more likely to use a modern reversible method than women in communities not served by the Programme (OR=1.50, 95% CI=1.04-2.16, p=0.031), even after controlling for various household and individual characteristics. The model of providing doorstep services through community-based female workers should remain central to achieving universal access to safe family planning methods by the end of the decade--the long-term objective of Pakistan's most recent population policy adopted in 2002. PMID- 15746221 TI - Quality medicines for the poor: experience of the Delhi programme on rational use of drugs. AB - Prior to 1994, most Delhi hospitals and dispensaries experienced constant shortages of essential medicines. There was erratic prescribing of expensive branded products, frequent complaints about poor drug quality and low patient satisfaction. Delhi took the lead in developing a comprehensive Drug Policy in 1994 and was the only Indian state to have such a comprehensive policy. The policy's main objective is to improve the availability and accessibility of quality essential drugs for all those in need. The Delhi Society for the Promotion of Rational Use of Drugs (DSPRUD), a non-governmental organization, worked in close collaboration with the Delhi Government and with universities to implement various components of the policy. The first Essential Drugs List (EDL) was developed, a centralized pooled procurement system was set up and activities promoting rational use of drugs were initiated. In 1997, the Delhi Programme was designated the INDIA-WHO Essential Drugs Programme by the World Health Organization. The EDL was developed by a committee consisting of a multidisciplinary group of experts using balanced criteria of efficacy, safety, suitability and cost. The first list contained 250 drugs for hospitals and 100 drugs for dispensaries; the list is revised every 2 years. The pooled procurement system, including the rigorous selection of suppliers with a minimum annual threshold turnover and the introduction of Good Manufacturing Practice inspections, resulted in the supply of good quality drugs and in holding down the procurement costs of many drugs. Bulk purchasing of carefully selected essential drugs was estimated to save nearly 30% of the annual drugs bill for the Government of Delhi, savings which were mobilized for procuring more drugs, which in turn improved availability of drugs (more than 80%) at health facilities. Further, training programmes for prescribers led to a positive change in prescribing behaviour, with more than 80% of prescriptions being from the EDL and patients receiving 70-95% of the drugs prescribed. These changes were achieved by changing managerial systems with minimal additional expenditure. The 'Delhi Model' has clearly demonstrated that such a programme can be introduced and implemented and can lead to a better use and availability of medicines. PMID- 15746222 TI - 20 best resources on globalization. PMID- 15746223 TI - Neuroimaging of cerebral venous thrombosis. AB - Early and accurate diagnosis of cerebral venous thrombosis (CVT) is possible with the help of computed tomography (CT) scan and magnetic resonance imaging (MRI). Empty delta sign on postcontrast CT is present in only up to 30% of the cases. The role of CT venography is not yet established, but it is emerging as an effective modality for diagnosis of CVT. T2* MRI sequence is superior to spin echo in detecting CVT and small hemor rhages. MR venography is considered the technique of choice for diagnosis and follow-up of CVT, but in certain cases, MRI could be superior as it shows the thrombus itself and not just the absence of signal as seen on MR venography. Diffusion-weighted imaging is a relatively new MRI technique that is extremely sensitive in detecting acute arterial strokes and can distinguish cytotoxic and vasogenic edema. The presence of hyperintense signal on diffusion-weighted imaging in the occluded veins or sinuses at the time of diagnosis may predict a low rate of vessel recanalization. Contrast angiography should be strongly considered if isolated cortical vein thrombosis is suspected. PMID- 15746224 TI - Neuroimaging determinants of cognitive performances in stroke associated with small vessel disease. AB - BACKGROUND AND PURPOSE: Controversies still exist as to the neuroimaging determinants of cognitive impairment in cerebral small vessel disease (SVD). The authors studied the neuroimaging correlates of cognitive performances among patients with stroke associated with SVD. METHODS: The authors per formed cerebral computed tomography, magnetic resonance imaging, and diffusion-weighted imaging among 74 consecutive patients admitted to the acute stroke unit because of stroke associated with SVD. They examined the association between cognitive performances and the following neuroimaging features: volume of white matter changes (WMC), multiplicity of lacunae, location of lacunae, total cerebral atrophy, and frontal and medial temporal lobe atrophy. RESULTS: Apart from age and education, univariate linear regression analyses revealed that WMC volume, presence of thalamic lacunae, cerebral atrophy, and left frontal lobe atrophy predicted performance on the Mini-Mental State Examination while WMC volume, presence of thalamic infarcts, cerebral atrophy, and frontal lobe atrophy of both sides predicted performance on the Mattis Dementia Rating Scale Initiation/Preservation subscale. In the multivariate analyses, education (R2=0.22, P<.001), left frontal lobe atrophy (R2=0.10, P=.004), and presence of thalamic lacunae (R2=0.04, P=.049) were found to predict performance on the Mini Mental State Examination while age (R2=0.23, P<.001) and presence of thalamic lacunae (R2=0.08, P=.011) were found to predict performance on the Mattis Dementia Rating Scale-Initiation/Preservation. CONCLUSIONS: Among patients with stroke associated with SVD, thalamic lacunae and frontal lobe atrophy are key determinants of cognitive performances. PMID- 15746225 TI - Bedside emergency transcranial Doppler diagnosis of severe carotid disease using orbital window examination. AB - BACKGROUND: Identifying internal carotid artery (ICA) stenosis in the acute stroke setting can provide clinically useful information. Transcranial Doppler (TCD) through the orbital window is an easy test to perform and to track and identify different vessels. Previous TCD studies have suggested that a reversed ophthalmic artery (OA) flow is a useful collateral pattern to predict ICA disease. The authors sought to evaluate the TCD orbital window for predicting cervical ICA (cICA) stenosis in the setting of acute stroke and TIA. METHOD: Power M-mode/TCD was performed in acute stroke and transient ischemic attack patients at 2 institutions. Each orbital window depth was detected on M-mode and evaluated for the direction of flow and resistance pattern. Gold standard for comparison was carotid evaluation using carotid duplex, computed tomography angiogram, or conventional angiography. The assessment of cICA disease was categorized by degree of stenosis or occlusion. RESULTS: A total of 216 transorbital exams were performed in 117 patients. Twenty-five cICA occlusions and 8 critical cICA stenoses (>or=95%) were identified by gold standard imaging. Reversed OA flow at 50 to 60 mm depth revealed high specificity (100%; confidence interval [CI], 97.6%-100.0%) and good sensitivity (75%; CI, 53.3%-90.2%) for identifying cICA occlusion or critical stenosis (>or=95%). Low pulsatility index (<1.2) and mean flow velocity (<15 cm/s) discriminated critical severe ICA stenosis or occlusion when OA flow was anterograde with good sensitivity (87.2%) and specificity (95.2%). CONCLUSION: The reversed OA sign at 50 to 60 mm depth is very specific for identifying cICA occlusion or critical stenosis. When OA flow is anterograde, a low mean flow velocity or pulsatility index is also useful to identify cICA critical stenosis or occlusion. PMID- 15746226 TI - Carbon monoxide brain toxicity: clinical, magnetic resonance imaging, magnetic resonance spectroscopy, and neuropsychological effects in 9 people. AB - Carbon monoxide (CO) exposure is a common cause of toxic brain damage, whereby effects range from transient neurological dysfunction to coma and death. A spectrum of severity of magnetic resonance imaging (MRI) findings after CO brain toxicity, including globus pallidus and white matter lesions, is well documented. Reports of MR spectroscopy (MRS) findings re main sparse. This article reports 9 people exposed to CO because of an apartment house's faulty gas heater. Four, with transient loss of consciousness after chronic moderate level CO exposure, suffered intellectual impairment without MRI abnormalities. The MRS of 1 individual demonstrated decreased n-acetyl aspartase in the basal ganglia, bilaterally. Of 5 exposed to high levels for about 12 hours, 1 died prior to clinical and/or MRI evaluation. One who suffered coma recovered but was lost to evaluation. Three, who were unconscious for hours to days, exhibited T2 MRI white matter signal abnormalities. MRS showed decreased basal ganglia n-acetyl aspartase in 2. One of these suffers a Parkinsonian syndrome. All 3 are intellectually impaired. This study demonstrates that although MRI and MRS are useful markers of CO-induced brain damage, they are not always sensitive to resultant intellectual dysfunction. PMID- 15746227 TI - Localized cerebral blood flow reductions in patients with heart failure: a study using 99mTc-HMPAO SPECT. AB - BACKGROUND AND PURPOSE: Reduced resting global cerebral blood flow has been previously detected in association with heart failure (HF), but it is not clear whether there are brain regions that could be specifically affected by those brain perfusion deficits. The authors used a fully automated, voxel-based image analysis method to investigate, across the entire cerebral volume, the presence of resting regional cerebral blood flow (rCBF) abnormalities in HF patients compared to healthy controls. METHODS: rCBF was evaluated with 99mTc-single photon emission computed tomography in 17 HF patients (New York Heart Association functional class II or III) and 18 elderly healthy volunteers. Voxel-based analyses of rCBF data were conducted using the statistical parametric mapping software. RESULTS: Significant rCBF reductions in HF patients relative to controls (P<.05, corrected for multiple comparisons) were detected in 2 foci, encompassing, respectively, the left and right precuneus and cuneus and the right lateral temporoparietal cortex and posterior cingulated gyrus. In the HF group, there was also a significant direct correlation between the degree of cognitive impairment as assessed using the Cambridge Mental Disorders of the Elderly Examination and rCBF on a voxel cluster involving the right posterior cingulate cortex and precuneus, located closely to the site where between-group rCBF differences had been identified. CONCLUSIONS: These preliminary findings indicate that posterior cortical areas of the brain may be particularly vulnerable to brain perfusion reductions associated with HF and suggest that functional deficits in these regions might be relevant to the pathophysiology of the cognitive impairments presented by HF patients. PMID- 15746228 TI - Dotlike hemosiderin spots are associated with past hemorrhagic strokes in patients with lacunar infarcts. AB - BACKGROUND AND PURPOSE: Dotlike hemosiderin spots ongradient-echo T2(*)-weighted magnetic resonance imaging of the brain have been histologically diagnosed as old microbleeds associated with small vessel disease (SVD). The authors hypothesize that the presence of many dotHSs may be correlated with the fragility of small vessels and the recurrence of SVD, including lacunar infarction and deep intracerebral hemorrhage (ICH). METHODS: To investigate how dotHSs are related to past history of SVD, the number of subcortical or deep dotHSs was investigated in 146 patients with lacunar infarctions (95 men, 51 women, age 38 to 90 [66.6+/ 9.4] years). They were divided into 2 subgroups according to history of deep ICHs or lacunar infarctions. The odds ratio (OR) for past history was estimated from logistic regression analyses with the number of subcortical or deep dotHSs as well as other factors. RESULTS: Of 146 patients with lacunar infarctions, 11 had past symptomatic ICHs and 19 had past symptomatic lacunar infarctions. An elevated rate of history of ICH was found for lacunar infarction patients with many deep dotHSs (>or=3; OR, 9.1; 95% confidence interval, 1.6-51, P=.015). However, history of lacunar infarction was not significantly associated with the number of subcortical or deep dotHSs. CONCLUSIONS: Our findings suggest that many deep dotHSs on T2(*)-weighted magnetic resonance imaging may be correlated with deep ICH-lacunar infarction type of SVD recurrence but not lacunar infarction lacunar infarction type. PMID- 15746229 TI - Diffusion-weighted magnetic resonance imaging of spinal infection and malignancy. AB - BACKGROUND AND PURPOSE: Pyogenic and tuberculous spondylitis can mimic malignancy. The purpose of this study was to deter mine the efficacy of diffusion weighted magnetic resonance imaging in differentiating spinal infection and malignancy. METHODS: Fifty-one consecutive patients with suspected spinal infection or malignancy were enrolled in the study. Apparent diffusion coefficients (ADCs) of paraspinal soft tissue mass and normal and abnormal vertebral bone marrow were determined on the diffusion-weighted magnetic resonance images of the spine. The mean ADCs of normal and abnormal vertebral bodies in patients with confirmed infection or malignancy were compared using nonparametric tests. RESULTS: ADCs of 69 tuberculous, 9 pyogenic, and 50 malignant vertebral marrow lesions were significantly higher than ADCs of normal marrow. ADCs of malignant bone marrow and 5 paraspinal soft tissue lesions were significantly lower than tuberculosis and pyogenic infection. There was no significant difference between the ADCs of 44 adult and 25 pediatric tuberculous bone lesions or between tuberculosis and pyogenic infection. Using the cutoff ADC of 1.02x10(-3)mm2/s for bone marrow, the sensitivity, specificity, and accuracy were 60.26%, 66.00%, and 62.50%, respectively, for distinguishing infection from malignancy. The sensitivity, specificity, and accuracy increased to 94.12%, 82.35%, and 90.20%, respectively, when the ADCs of associated soft tissue lesions were higher than 1.17x10(-3)mm2/s. CONCLUSIONS: Diffusion-weighted magnetic resonance imaging has limited usefulness for differentiating spinal infection and malignancy. PMID- 15746230 TI - Nonlinear responses within the medial prefrontal cortex reveal when specific implicit information influences economic decision making. AB - BACKGROUND AND PURPOSE: The authors used functional magnetic resonance imaging (fMRI) to investigate how individual economic decisions are influenced by implicit memory contributions. METHODS: Twenty-two participants were asked to make binary decisions between different brands of sensorily nearly undistinguishable consumer goods. Changes of brain activity comparing decisions in the presence or absence of a specific target brand were detected by fMRI. RESULTS: Only when the tar get brand was the participant's favorite one did the authors find reduced activation in the dorsolateral prefrontal, posterior parietal, and occipital cortices and the left premotor area (Brodmann areas [BA] 9, 46, 7/19, and 6). Simultaneously, activity was increased in the inferior precuneus and posterior cingulate (BA 7), right superior frontal gyrus (BA 10), right supramarginal gyrus (BA 40), and, most pronounced, in the ventromedial prefrontal cortex (BA 10). CONCLUSIONS: For products mainly distinguishable by brand information, the authors revealed a nonlinear winner-take-all effect for a participant's favorite brand characterized, on one hand, by reduced activation in brain areas associated with working memory and reasoning and, on the other hand, increased activation in areas involved in processing of emotions and self reflections during decision making. PMID- 15746231 TI - Magnetic resonance imaging protocols for cervical disc disease: what is your neighbor up to? AB - BACKGROUND AND PURPOSE: To summarize the prevalence of particular protocols used for routine magnetic resonance imaging of the cervical spine. METHODS: Centers nationwide were asked to submit routine protocols for degenerative disease. Cases with a history of demyelinating disease or suspected cord lesions were excluded. The pulse sequences and scanning planes were tabulated for 105 centers. For the last 53 centers studied, axial gradient recalled echo sequences were subdivided into 2-dimensional and 3-dimensional Fourier transform. RESULTS: There were 21 different combinations. A protocol consisting of sagittal T1+sagittal T2+axial gradient recalled echo T2 was used by 48% of centers. Two sagittal and 2 axial series in any combination was used in 30% (32/105 centers). One center used gradient recalled echo images exclusively, consisting of T1, proton density, and T2-weighted images, which included axial and sagittal planes. CONCLUSION: Sagittal T1+sagittal T2+axial gradient recalled echo T2 is by far the most prevalent protocol used for imaging the cervical spine. PMID- 15746232 TI - Reversible posterior leukoencephalopathy, cerebral vasoconstriction, and strokes after intravenous immune globulin therapy in guillain-barre syndrome. AB - The authors report a patient with Guillain-Barre syndrome who developed acute hypertension, reversible posterior leukoencephalopathy syndrome, ischemic and hemorrhagic strokes, and reversible cerebral arterial vasoconstriction shortly after initiating intravenous immune globulin therapy. Possible interrelationships and mechanisms of these complications are discussed. PMID- 15746233 TI - Magnetic resonance imaging and diffusion-weighted imaging changes after hypoglycemic coma. AB - The authors report a case of severe hypoglycemic encephalopathy in an elderly patient. The magnetic resonance images showed bilateral cortical signal changes and basal ganglia lesions, which spared the thalami. The lesions were bright on fluid-attenuated inversion recovery and diffusion-weighted images and dark on the apparent diffusion coefficient map, being more conspicuous on the diffusion weighted images than on the fluid-attenuated inversion recovery images. A literature review of the imaging features and pathophysiological mechanism in comparison with those of hypoxic ischemic injury is discussed. PMID- 15746234 TI - Direct infiltration of brainstem glioma along the cranial nerves. AB - The authors describe a case of a low-grade brainstem glioma extending along the cranial nerves without any evidence of leptomeningeal spread. The tumor extended directly along the VII-VIIIth cranial nerve complex and also along the trigeminal nerve, which is quite an unusual characteristic of the glial tumors. PMID- 15746235 TI - Expanding the spectrum of acquired cerebral hemiatrophy. AB - The authors report a 50-year-old man with progressive left hemispheric atrophy, cognitive decline, infrequent seizures, and spells suggestive of complicated migraine. They discuss the association between migraine and cerebral hemiatrophy and suggest that the spectrum of acquired cerebral hemiatrophy may be broader than what has been commonly recognized. PMID- 15746236 TI - Evidence of acute ischemic tissue change in transient global amnesia in magnetic resonance imaging: case report and literature review. AB - Transient global amnesia is a benign syndrome of sudden-onset alteration of behavior with temporary dysfunction of anterograde and recent retrograde memory. Its neural substrates remain uncertain. Possible causes include ischemia, migraine, and epilepsy. The authors report a case of a 62-year-old man with a transient attack of memory disturbance, suggestive of transient global amnesia, in which magnetic resonance imaging performed 48 hours after onset showed left mesial temporal lobe signal changes on diffusion-weighted imaging and fluid attenuated inversion recovery images. The findings and a literature review lend further support to the ischemic pathogenesis of transient global amnesia as a possible etiology, and underscore the role of diffusion-weighted imaging in the diagnosis of this condition. PMID- 15746237 TI - Magnetic resonance images on Marburg variant. PMID- 15746240 TI - Physical limits to spatial resolution of optical recording: clarifying the spatial structure of cortical hypercolumns. AB - Neurons in macaque primary visual cortex are spatially arranged by their global topographic position and in at least three overlapping local modular systems: ocular dominance columns, orientation pinwheels, and cytochrome oxidase (CO) blobs. Individual neurons in the blobs are not tuned to orientation, and populations of neurons in the pinwheel center regions show weak orientation tuning, suggesting a close relation between pinwheel centers and CO blobs. However, this hypothesis has been challenged by a series of optical recording experiments. In this report, we show that the statistical error associated with photon scatter and absorption in brain tissue combined with the blurring introduced by the optics of the imaging system has typically been in the range of 250 microm. These physical limitations cause a systematic error in the location of pinwheel centers because of the vectorial nature of these patterns, such that the apparent location of a pinwheel center measured by optical recording is never (on average) in the correct in vivo location. The systematic positional offset is approximately 116 microm, which is large enough to account for the claimed misalignment of CO blobs and pinwheel centers. Thus, optical recording, as it has been used to date, has insufficient spatial resolution to accurately locate pinwheel centers. The earlier hypothesis that CO blobs and pinwheel centers are coterminous remains the only hypothesis currently supported by reliable observation. PMID- 15746241 TI - Genomic imprinting of XX spermatogonia and XX oocytes recovered from XX<-->XY chimeric testes. AB - We produced XX<-->XY chimeras by using embryos whose X chromosomes were tagged with EGFP (X*), making the fluorescent green female (XX*) germ cells easily distinguishable from their nonfluorescent male (XY) counterparts. Taking advantage of tagging with EGFP, the XX* "prospermatogonia" were isolated from the testes, and the status of their genomic imprinting was examined. It was shown that these XX cells underwent a paternal imprinting, despite their chromosomal constitution. As previously indicated in sex-reversal XXsxr testes, we also found a few green XX* germ cells developed as "eggs" within the seminiferous tubules of XX*<-->XY chimeric testes. These cells were indistinguishable from XX* prospermatogonia at birth but resumed oogenesis in a testicular environment. The biological nature of the "testicular eggs" was examined by recovering the eggs from chimeric testes. The testicular eggs not only formed an egg-specific structure, the zona pellucida, but also were able to fuse with sperm. The collected testicular eggs were indicated to undergo maternal imprinting, despite the testicular environment. The genomic imprinting did not always follow the environmental conditions of where the germ cells resided; rather, it was defined by the sex that was chosen by the germ cells at early embryonic stage. PMID- 15746242 TI - The central and basolateral nuclei of the amygdala exhibit opposite diurnal rhythms of expression of the clock protein Period2. AB - There is considerable evidence that circadian rhythms in mammals can be modulated by emotional state, but how emotional state modulates specific circadian outputs is poorly understood. We analyzed the expression of the circadian clock protein Period2 (PER2) in three regions of the limbic forebrain known to play key roles in emotional regulation, the central nucleus of the amygdala (CEA), the basolateral amygdala (BLA), and the dentate gyrus (DG). We report here that cells in all three regions exhibit daily rhythms in expression of PER2 that are under the control of the master clock, the suprachiasmatic nucleus (SCN). The rhythm in the CEA and the rhythms in the BLA and DG are diametrically opposite in phase and are differentially affected by adrenalectomy. Adrenalectomy completely abolished the PER2 rhythm in the CEA but had no effect on the PER2 rhythms in the BLA and DG. We previously reported a rhythm in PER2 expression in the oval nucleus of the bed nucleus of the stria terminalis that is identical in phase and sensitivity to adrenalectomy to that found in the CEA. Together, these findings show that key structures of the limbic forebrain exhibit daily oscillations in clock gene expression that are controlled not only by input from the SCN but, importantly, by hormonal and neurochemical changes that normally accompany motivational and emotional states. Thus, cells within these areas are strategically positioned to integrate the inputs from the SCN and emotional states to modulate circadian rhythms downstream from the SCN clock. PMID- 15746244 TI - CD83 localization in a recycling compartment of immature human monocyte-derived dendritic cells. AB - Dendritic cells (DC) change their phenotype and functional properties during maturation. CD83 cell surface expression is induced on mature DC (mDC). In this study, we investigated intracellular CD83 localization and transport in human monocyte-derived DC. The enhanced level of CD83 cell surface expression in mDC resulted predominantly from increased protein synthesis, and in addition from regulated intracellular transport of CD83 protein. An internal pool of CD83 protein is present in immature DC (iDC). Although CD83 protein in iDC and in mDC was localized in the Golgi compartment and in recycling endosomes, only in mature cells did CD83 co-localize with MHC class II molecules in endocytic vesicles. CD83 cell surface expression on iDC was induced by inhibition of endocytosis. This result could be explained by CD83 cycling between endosomes and the cell surface in iDC. The mDC also rapidly internalized membrane-bound CD83 protein. Furthermore, a thiol protease inhibitor and specific cathepsin inhibitors impaired CD83 up-regulation in DC, indicating a role of endosomal proteases in the maturation-induced exposure of CD83 on the plasma membrane. PMID- 15746243 TI - Early lymphoid progenitors in mouse and man are highly sensitive to glucocorticoids. AB - Glucocorticoids are extensively used in anti-inflammatory therapy and may contribute to the normal regulation of lymphopoiesis. This study utilized new information about the early stages of lymphopoiesis in mouse and man to determine precisely which cell types are hormone sensitive. Cycling B lineage precursors were depleted in dexamethasone-treated mice, while mature, non-dividing CD45R(Hi) CD19(Hi) lymphocytes, myeloid progenitors and stem cells with the potential for lymphocyte generation on transplantation were spared. Lineage marker-negative (Lin(-)) IL-7R(+) Flk-2(+) pro-lymphocytes also declined, but not as rapidly as the terminal deoxynucleotidyl transferase-positive cells within an early Lin(-) c kit(Hi) Sca-1(Hi) fraction of bone marrow. Hormone-sensitive cells with additional properties of early lymphoid progenitors (ELP) were identified within the same Lin(-) c-kit(Hi) Sca-1(Hi) subset using human mu transgenic mice and recombination-activating gene 1 (RAG1)/green fluorescent protein knock-in animals. Furthermore, cells with a recent history of RAG1 expression were more glucocorticoid sensitive than mature lymphocytes in marrow and spleen. Lymphocyte progenitors in mice bearing a human bcl-2 transgene were protected from dexamethasone treatment. However, isolated progenitors from either wild-type or bcl-2 transgenic mice were directly sensitive to the hormone in stromal cell-free cultures, suggesting that additional factors must determine vulnerability to glucocorticoids. B lineage lymphocyte precursors were found to be abnormally elevated in the bone marrow of adrenalectomized or RU486-treated mice. This suggests that glucocorticoids may normally contribute to steady-state regulation of lymphopoiesis. Finally, parallel studies revealed that the earliest events in human lymphopoiesis are susceptible to injury during glucocorticoid therapy. PMID- 15746245 TI - Pertussis toxin B-oligomer inhibits HIV infection and replication in hu-PBL-SCID mice. AB - Bordetella pertussis toxin B-oligomer (PTX-B) has been shown to inhibit HIV infection and replication in vitro. The potential anti-viral effect of PTX-B was tested here in an in vivo surrogate model of HIV infection, i.e. SCID mice reconstituted with human peripheral blood leukocytes (PBL) (hu-PBL-SCID) and infected with a CCR5-dependent (R5) HIV-1 strain. SCID mice inoculated intra peritoneal (i.p.) with PTX-B and then infected with the R5 strain SF-162 were sacrificed 7 days later and analyzed for human PBL (hu-PBL) lymphoid tissue reconstitution, infection of hu-PBL, plasma viremia and viral rescue from ex vivo cultivated i.p. hu-PBL. Unlike mice treated with 500 ng per animal of PTX-B showing no evidence of viral inhibition, daily administration of PTX-B (50 ng per mouse) strongly inhibited virus infection and replication, as determined by undetectable viremia, absence of infected hu-PBL and lack of rescue of infectious HIV in most animals. Furthermore, PTX-B injection 2 h before and twice after infection prevented HIV-1 infection and replication in all (10/10) tested animals. Thus, PTX-B potently inhibited virus infection and replication in hu-PBL SCID mice, supporting the hypothesis that it may represent a new pharmacological agent against HIV-1 infection. PMID- 15746247 TI - Sequence requirements for oligodeoxyribonucleotide inhibitory activity. AB - Inhibitory oligonucleotides (IN-ODN) differing from stimulatory CpG ODN (ST-ODN) by as few as two bases can block ST-ODN-induced proliferation, apoptosis protection and IL-6 secretion in B lymphocytes and the production of IL-12p40 by non-B cells. The main objective of this study was to determine the ODN sequence requirements for inhibition in mice. Starting with a strongly inhibitory 15-mer prototype phosphorothioate sequence, we tested the 60 sequences that differed from the prototype by one base, revealing the three areas that are critical for activity. Between these areas were the spacer sequences where base composition mattered little, but the number of bases was important. Truncation of three bases at the 3' end of the 15-mer and one at the 5' end was tolerated with minimal loss of activity. This approach yielded an 'optimal' sequence of 5' CC x notC notC xxGGGx or CC x notC notC xGGGxx 3', where x is any base. The sequence requirements for optimal inhibition of B cell responses to Type B (K) ODN and mixed splenocyte IL-12p40 responses to Type A (D) ODN were strikingly similar. Inhibition of ST-ODN by IN-ODN was competitive. A hypothetical model of the ODN binding site is proposed. Synthetic IN-ODN with the sequence characteristics defined here should provide antidotes for excessive innate reactions to DNA. PMID- 15746246 TI - Soluble CD100 functions on human monocytes and immature dendritic cells require plexin C1 and plexin B1, respectively. AB - CD100 represents the first semaphorin described in the immune system. It is expressed as a 300-kDa homodimer at the surface of most hematopoietic cells, but is also found in a soluble form following a proteolytic cleavage upon cell activation. We herein established that soluble CD100 (sCD100) impaired the migration of human monocytes and immature dendritic cells (DCs), but not of mature DCs. Performing competition assays, we identified plexin C1 (VESPR/CD232) as being involved in sCD100-mediated effects on human monocytes. Interestingly, we observed a complete down-regulation of plexin C1 expression during the in vitro differentiation process of monocytes to immature DCs, while concomitantly the surface expression of plexin B1 was induced. The latter receptor then binds sCD100 on immature DCs, mediating its inhibitory effect on cell migration. Finally, we showed that sCD100 modulated the cytokine production from monocytes and immature DCs. Together these results suggest that sCD100 plays a critical role in the regulation of antigen-presenting cell migration and functions via a tightly regulated process of receptor expression. PMID- 15746248 TI - Genomic and functional delineation of dendritic cells and memory T cells derived from grass pollen-allergic patients and healthy individuals. AB - Dendritic cells (DCIs) possess a potent ability to modulate and activate specific T-cell responses to allergens, which play a pivotal role in allergic inflammation by secreting cytokines and other mediators. However, the molecular mechanisms by which allergen-challenged DCs regulate specific T-cell responses are still not well characterized. This study aims at elucidating the molecular mechanisms underlying the DC-T-cell interaction during an allergic immune response to grass pollen, using a genomic and functional approach. Transcriptional analysis was performed on grass allergen Phleum pratense-stimulated DCs and on autologous memory CD4(+) T cells co-cultured with allergen-challenged DCs from healthy and allergic donors. DCs from the allergic donors were potent inducers of T-cell proliferation and T(h)2 polarization, as demonstrated by high IL-4, IL-5 and IL 13, and low IFN-gamma production. A gradual up-regulation of activation markers on both DCs and T cells was evident during the co-culture period, demonstrating an educational element of the DC-T-cell interaction. The global transcriptional analysis revealed a differential gene regulation in DCs and T cells derived from allergic donors after stimulation with allergen, as compared with the healthy donors. Peripheral memory CD4(+) T cells from healthy and allergic donors also responded differently after stimulation with allergen-loaded DCs with respect to cytokine production, proliferation, surface marker expression and gene transcription. We found up-regulated genes involved in T(h)2 cell biology, such as genes important for homing, adhesion, signaling and transcription, in addition to genes previously not described in the context of allergy. The panel of differentially expressed genes in the allergic group will form the basis for an increased understanding of the molecular mechanisms in allergy. PMID- 15746249 TI - Tumor necrosis factor-{alpha} decreases Akt protein levels in 3T3-L1 adipocytes via the caspase-dependent ubiquitination of Akt. AB - TNF-alpha is a mediator of insulin resistance in sepsis, obesity, and type 2 diabetes and is known to impair insulin signaling in adipocytes. Akt (protein kinase B) is a crucial signaling mediator for insulin. In the present study we examined the posttranslational mechanisms by which short-term (<6-h) exposure of 3T3-L1 adipocytes to TNF-alpha decreases Akt levels. TNF-alpha treatment both increased the ubiquitination of Akt and decreased its protein level. The decrease in protein was associated with the presence of an (immunoreactive) Akt fragment after TNF-alpha treatment, indicative of Akt cleavage. The broad-spectrum caspase inhibitor t-butoxycarbonyl-Asp(O-Me)-fluoromethyl ketone markedly suppressed these effects of TNF-alpha. The caspase-6 inhibitor Z-Val-Glu(OMe)-Ile-Asp(OMe) CH(2)F potently suppressed Akt ubiquitination, degradation, and fragment formation, whereas the proteasome inhibitor Z-Leu-Leu-Leu-CHO modestly attenuated the decline in Akt levels. Exposure to TNF-alpha also enhanced the association of Akt with an E3 ligase activity. Adipocytes preexposed to TNF-alpha for 5 h and then stimulated with insulin for 30 min exhibited decreased levels of Akt, phosphorylated Akt, as well as phosphorylated Mdm2, which is a known direct substrate of Akt, and glucose uptake. Caspase inhibition attenuated these inhibitory effects of TNF-alpha. Collectively, our results suggest that TNF-alpha induces the caspase-dependent degradation of Akt via the cleavage and ubiquitination of Akt, which results in its degradation through the 26S proteasome. Furthermore, the caspase- and proteasome-mediated degradation of Akt due to TNF-alpha exposure leads to impaired Akt-dependent insulin signaling in adipocytes. These findings expand the mechanism by which TNF-alpha impairs insulin signaling. PMID- 15746250 TI - Leptin orally supplied to neonate rats is directly uptaken by the immature stomach and may regulate short-term feeding. AB - Although leptin is a hormone mainly produced by the adipose tissue, it is also produced by the gastric mucosa and the mammary epithelium and is present in maternal milk. The effects of milk leptin on the neonate are not known. The purpose of the investigation was to evaluate the short-term effects of the administration of a single oral dose of leptin on 4-d-old rats as well as the effects of chronic supplementation during the lactation period with a daily oral dose of leptin (equivalent to 5 times the amount of leptin ingested normally from maternal milk during the suckling period) on body weight, the gastric leptin system, gastric food content, and thermogenic capacity. Our results show that the administration of a single oral dose of 4 ng of leptin to 4-d-old rats produces a short-term increase in leptin levels in the stomach and serum and a decrease in the weight of the gastric contents. Pups treated with a daily oral dose of leptin during the whole lactation period showed, at the end of the suckling period, compared with controls, lower gastric contents, lower leptin production by the stomach and the sc adipose tissue, and lower thermogenic capacity in brown adipose tissue. We conclude that oral leptin is absorbed by the immature gastric epithelium of the neonate, and this leptin exerts clear biological effects, down regulating endogenous leptin production and playing a potential role in the short term control on food intake during the lactation period. PMID- 15746251 TI - The reciprocal switching of two thyroid hormone-activating and -inactivating enzyme genes is involved in the photoperiodic gonadal response of Japanese quail. AB - The molecular mechanisms underlying photoperiodic time measurement are not well understood in any organism. Relatively recently, however, it has become clear that thyroid hormones play an important role in photoperiodism, and in a previous study we reported that long daylengths in Japanese quail increase hypothalamic levels of T(3) and of the thyroid hormone-activating enzyme, type 2 iodothyronine deiodinase. The present study extends these observations to measure gene levels of the thyroid hormone-inactivating enzyme, type 3 deiodinase. Levels decreased after exposure to long days, but increased under short days. Changes in the two genes were then analyzed during the precisely timed photoinduction that occurs in quail exposed to a single long day. The two gene switches are the earliest events yet recorded in the photoinduction process, and overall, these reciprocal changes offer the potential to regulate active brain thyroid hormone concentrations rather precisely at the site in the brain where photoinduction is triggered. PMID- 15746253 TI - SRC homology-2-containing protein tyrosine phosphatase-1 restrains cell proliferation in human medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) is a rare tumor originating from thyroid parafollicular C cells, where, in the inherited form, constitutive activation of the RET protooncogene is responsible for unrestrained cell proliferation. We previously demonstrated that somatostatin (SRIF) reduces cell growth in the human MTC cell line TT, which expresses all SRIF receptor (SSTR) subtypes and responds differently to selective SSTR agonists. The antiproliferative mechanism of SRIF and its analogs in MTC is still unclear. Src homology-2-containing protein tyrosine phosphatase-1 (SHP-1), a cytoplasmic protein tyrosine phosphatase (PTP), is activated by somatotropin release-inhibiting factor and reduces mutated RET autophosphorylation in a heterologous system. In this study, we explore the role of PTP activation, in particular of SHP-1, in TT cells, where RET is constitutively activated. In TT cells, SRIF stimulated the PTP activity of SHP-1, which was associated with proliferation inhibition and with reduction in the MAPK pathway activation. Blockade of PTP activity with sodium orthovanadate induced cell proliferation and MAPK phosphorylation and blunted the inhibitory effects of SRIF. Moreover, SHP-1 associates with SSTR2 depending on its activation. By using a MAPK kinase inhibitor, we demonstrated that TT cell growth depends on MAPK pathway activation. Furthermore, in TT cells overexpressing SHP-1, cell proliferation and MAPK signaling were strongly down-regulated, whereas in TT cells transfected with a dominant negative form of SHP-1, cell proliferation and MAPK signaling were markedly induced. Our data demonstrate that SRIF inhibitory effects on TT cell proliferation are mediated, at least in part, by SHP-1, which acts through a MAPK-dependent mechanism. PMID- 15746252 TI - Astrocyte-derived transforming growth factor-{beta} mediates the neuroprotective effects of 17{beta}-estradiol: involvement of nonclassical genomic signaling pathways. AB - 17beta-Estradiol (E2) and selective estrogen receptor modulators (SERMs), such as tamoxifen, mediate numerous effects in the brain, including neurosecretion, neuroprotection, and the induction of synaptic plasticity. Astrocytes, the most abundant cell type in the brain, influence many of these same functions and thus may represent a mediator of estrogen action. The present study examined the regulatory effect and underlying cell signaling mechanisms of E2-induced release of neurotropic growth factors from primary rat cortical astrocyte cultures. The results revealed that E2 (0.5, 1, and 10 nm) and tamoxifen (1 mum) increased both the expression and release of the neuroprotective cytokines, TGF-beta1 and TGF beta2 (TGF-beta), from cortical astrocytes. The stimulatory effect of E2 was attenuated by the estrogen receptor (ER) antagonist, ICI182,780, suggesting ER dependency. The effect of E2 also appeared to involve mediation by the phosphotidylinositol 3-kinase (PI3K)/Akt signaling pathway, because E2 rapidly induced Akt phosphorylation, and pharmacological or molecular inhibition of the PI3K/Akt pathway prevented E2-induced release of TGF-beta. Additionally, the membrane-impermeant conjugate, E2-BSA, stimulated the release of TGF-beta, suggesting the potential involvement of a membrane-bound ER. Finally, E2, tamoxifen, and E2-BSA were shown to protect neuronal-astrocyte cocultures from camptothecin-induced neuronal cell death, effects that were attenuated by ICI182,780, Akt inhibition, or TGF-beta immunoneutralization. As a whole, these studies suggest that E2 induction of TGF-beta release from cortical astrocytes could provide a mechanism of neuroprotection, and that E2 stimulation of TGF-beta expression and release from astrocytes occurs via an ER-dependent mechanism involving mediation by the PI3K/Akt signaling pathway. PMID- 15746254 TI - Isolation and identification of histone H3 protein enriched in microvesicles secreted from cultured sebocytes. AB - Secretion of microvesicles, defined as sebosomes, containing lipid particles were discovered for the first time in cultured sebocytes. After reaching confluency, hamster-cloned sebocytes released bubble-like microvesicles with a diameter range of 0.5-5.0 microm. They had a complex structure containing multiple Oil Red O stainable particles. The lipid components of the microvesicles were large amounts of squalene both of hamster-cloned and rat primary cultured sebocytes. The microvesicles contained a concentrated 17-kDa cationic protein, which was soluble in sulfate buffer including Nonidet P-40 at pH 1.5. As the protein bound tightly to heparin-Sepharose and eluted with 1.5 M NaCl, it was further purified from a SDS-PAGE gel. Peptide sequencing identified the protein to be histone H3. Polyclonal antibodies against the purified protein detected the antigen in the microvesicles both in the hamster-cloned and rat primary cultured sebocytes. The antibodies demonstrated a distribution of the protein within the nucleus, cytoplasm, and precursor microvesicles. When a gene construct encoding histone H3 enhanced green fluorescent protein was transfected to the sebocytes, fluorescence of the fusion proteins was detected within both the nucleus and the precursor microvesicles of the cytoplasm. The distribution of heparan sulfate was evident in the microvesicles, and it suggested the possibility that the histone H3 protein was recruited and then condensed to the secreted microvesicles by the molecules. In addition, the 14-3-3 protein, which was detected in the microvesicles, also may help incorporate the histone H3 protein in the microvesicles because it can bind to both histone and lipid particles. PMID- 15746255 TI - Estrogen selectively up-regulates the phospholipid hydroperoxide glutathione peroxidase in the oviducts. AB - The oviduct plays a crucial role in mammalian reproduction by providing an optimal environment for the final maturation and transport of gametes, fertilization, and early embryonic development. It is now recognized that these reproductive events in vitro can be either negatively or positively affected by reactive oxygen species such as hydrogen peroxide and lipid hydroperoxides. In the current study, we analyzed the expression of the phospholipid hydroperoxide glutathione peroxidase (PHGPx or GPx-4), a selenoenzyme that directly reduces membrane-bound lipid hydroperoxides in the bovine oviduct. Using in situ hybridization, we demonstrated that GPx-4 expression is almost restricted to the oviductal luminal epithelium in contrast to GPx-1, which is widely distributed, and GPx-2 and -3, which are mainly detected in the epithelial cells and lamina propria. Interestingly, real-time quantitative RT-PCR analysis showed that GPx-4 expression was highest during the follicular and postovulatory phases. In addition, GPx-4 expression was highest in the isthmus proximal to the dominant follicle during the follicular stage and remained high during the postovulatory period. This increased in expression of GPx-4 corresponded to increased GPx-4 enzymatic activity. Based on intrauterine infusion of estradiol, we determined that the increase in expression and activity of GPx-4 is estrogen mediated. This work clearly demonstrates that GPx-4 gene expression is influenced by the proximity of the dominant follicle in the oviduct in vivo. We propose that GPx-4 has an important role in the physiological control of peroxide tone in the bordering cells of the oviductal lumen. PMID- 15746256 TI - Inverse shift in circulating corticosterone and leptin levels elevates hypothalamic deiodinase type 2 in fasted rats. AB - During food deprivation, plasma T(4) and T(3) levels are decreased. Under this metabolic condition, hypothalamic deiodinase type 2 (D2) activity and mRNA levels are elevated, whereas TRH mRNA levels are suppressed. Systemic T(4) administration does not reverse these hypothalamic changes. The mechanism(s) that underlies this paradoxical regulation of D2 during fasting is unknown. We hypothesize that leptin and/or glucocorticoids play a role in these mechanisms, and their interactions may be an important regulator of the hypothalamic pituitary-thyroid axis. Thus, we assessed the effects of these hormones on D2 activity levels of food-deprived as well as fed animals using enzyme activity measurements. In food-deprived animals, corticosterone replacement reversed the inhibitory effect of adrenalectomy (ADX) on D2 induction, whereas ADX and ADX plus corticosterone replacement did not significantly affect D2 activity levels in rats fed ad libitum. Leptin administration to fed animals did not change D2 activity, whereas in fasted rats, leptin decreased D2 activity by reducing corticosterone plasma levels. When leptin was administered to fasted animals that were either ADX or ADX plus corticosterone treated at a high dose, D2 activity did not increase. Our results show that during fasting, diminishing leptin levels play a permissive role to enable glucocorticoid-induced up-regulation of D2. Thus, our observations suggest that appropriate induction of D2 activity during negative energy balance is dependent upon both leptin and glucocorticoid signaling. PMID- 15746257 TI - Ectoderm-targeted overexpression of the glucocorticoid receptor induces hypohidrotic ectodermal dysplasia. AB - Hypohidrotic ectodermal dysplasia is a human syndrome defined by maldevelopment of one or more ectodermal-derived tissues, including the epidermis and cutaneous appendices, teeth, and exocrine glands. The molecular bases of this pathology converge in a dysfunction of the transcription factor nuclear factor of the kappa enhancer in B cells (NF-kappaB), which is essential to epithelial homeostasis and development. A number of mouse models bearing disruptions in NF-kappaB signaling have been reported to manifest defects in ectodermal derivatives. In ectoderm targeted transgenic mice overexpressing the glucocorticoid receptor (GR) [keratin 5 (K5)-GR mice], the NF-kappaB activity is greatly decreased due to functional antagonism between GR and NF-kappaB. Here, we report that K5-GR mice exhibit multiple epithelial defects in hair follicle, tooth, and palate development. Additionally, these mice lack Meibomian glands and display underdeveloped sweat and preputial glands. These phenotypic features appear to be mediated specifically by ligand-activated GR because the synthetic analog dexamethasone induced similar defects in epithelial morphogenesis, including odontogenesis, in wild-type mice. We have focused on tooth development in K5-GR mice and found that an inhibitor of steroid synthesis partially reversed the abnormal phenotype. Immunostaining revealed reduced expression of the inhibitor of kappaB kinase subunits, IKKalpha and IKKgamma, and diminished p65 protein levels in K5-GR embryonic tooth, resulting in a significantly reduced kappaB-binding activity. Remarkably, altered NF-kappaB activity elicited by GR overexpression correlated with a dramatic decrease in the protein levels of DeltaNp63 in tooth epithelia without affecting Akt, BMP4, or Foxo3a. Given that many of the 170 clinically distinct ectodermal dysplasia syndromes still remain without cognate genes, deciphering the molecular mechanisms of this mouse model with epithelial NF kappaB and p63 dysfunction may provide important clues to understanding the basis of other ectodermal dysplasia syndromes. PMID- 15746258 TI - Dual regulatory effects of orexins on sympathetic nerve activity innervating brown adipose tissue in rats. AB - This study examined how orexin regulates the activity of the sympathetic nerves that innervate brown adipose tissue (BAT) in rats. Infusion of orexin A at a dose of 0.3 nmol into the third cerebral ventricle decreased BAT sympathetic nerve activity, compared with the effect of PBS (P < 0.05), whereas infusion of orexin B at the same dose caused a significant increase (P < 0.05). Pretreatment with a third cerebral ventricle injection of 2.24 micromol/kg alpha fluoromethylhistidine, an irreversible inhibitor of the histamine-synthesizing enzyme histidine decarboxylase, attenuated the orexin B-induced response of BAT sympathetic nerve activity, but not that induced by orexin A. These results indicate that orexins may regulate both BAT energy expenditure and thermogenesis through their dual effects on sympathetic nerve activity. In particular, orexin B regulates BAT sympathetic nerve activity via neuronal histamine in the hypothalamus. PMID- 15746259 TI - Developmental changes in the pattern of ghrelin's acyl modification and the levels of acyl-modified ghrelins in murine stomach. AB - Ghrelin is an acylated peptide hormone secreted primarily from endocrine cells in the stomach. The major active form of ghrelin is a 28-amino acid peptide with an n-octanoyl modification at Ser(3) (n-octanoyl ghrelin), which is essential for its activity. In addition to n-octanoyl ghrelin, other forms of ghrelin peptide exist, including des-acyl ghrelin, which lacks an acyl modification, and other minor acylated ghrelin species, such as n-decanoyl ghrelin, whose Ser(3) residue is modified by n-decanoic acid. Multiple reports have identified various physiological functions of ghrelin. However, until now, there have been no reports that explore the process of ghrelin acyl modification, and only a few studies have compared the levels of des-acyl, n-octanoyl, and/or other minor populations of acylated ghrelin peptides. In this study we report that the amount of n-octanoyl ghrelin in murine stomachs increases gradually during the suckling period to a maximal level at 3 wk of age and falls sharply after the initiation of weaning. However, the concentration (picomoles per milligram of wet weight tissue) of total ghrelin, which includes des-acyl and all acylated forms of ghrelin peptides with intact C termini in murine stomach, remains unchanged across this suckling-weaning transition. Prematurely weaned mice exhibited a significant decrease in the amount of n-octanoyl or n-decanoyl ghrelin in the stomach. Orally ingested glyceryl trioctanoate, a medium-chain triacylglyceride rich in milk lipids, significantly increased the level of n-octanoyl-modified ghrelin in murine stomach. Fluctuations in the proportion of this biologically active, acyl-modified ghrelin could contribute to or be influenced by the change in energy metabolism during the suckling-weaning transition. PMID- 15746260 TI - Joint and bone disease in mucopolysaccharidoses VI and VII: identification of new therapeutic targets and biomarkers using animal models. AB - The mucopolysaccharidoses (MPS) are inherited metabolic disorders resulting from the defective catabolism of glycosaminoglycans. In this report, we find that the stimulation of MPS connective tissue cells by the inflammatory cytokines causes enhanced secretion of several matrix-degrading metalloproteinases (MMPs). In addition, expression of tissue inhibitor of metalloproteinase-1 was elevated, consistent with the enhanced MMP activity. These findings were not restricted to one particular MPS disorder or species, and are consistent with previous observations in animal models with chemically induced arthritis. Bromodeoxyuridine incorporation studies also revealed that MPS chondrocytes proliferated up to 5-fold faster than normal chondrocytes, and released elevated levels of transforming growth factor-beta, presumably to counteract the marked chondrocyte apoptosis and matrix degradation associated with MMP expression. Despite this compensatory mechanism, studies of endochondral ossification revealed a reduction in chondro-differentiation in the growth plates. Thus, although MPS chondrocytes grew faster, most of the newly formed cells were immature and could not mineralize into bone. Our studies suggest that altered MMP expression, most likely stimulated by inflammatory cytokines and nitric oxide, is an important feature of the MPS disorders. These data also identify several proinflammatory cytokines, nitric oxide, and MMPs as novel therapeutic targets and/or biomarkers of MPS joint and bone disease. This information should aid in the evaluation of existing therapies for these disorders, such as enzyme replacement therapy and bone marrow transplantation, and may lead to the development of new therapeutic approaches. PMID- 15746261 TI - Early inhaled nitric oxide improves oxidative balance in very preterm infants. AB - Inhaled nitric oxide (iNO) improves oxygenation in premature infants, but concern has been raised about its potential oxidative toxicity. We designed this study to assess the oxidative balance in premature infants who were exposed to low dose iNO and the relationship with their clinical outcome on day 28 of life. A total of 274 infants who were <32 wk gestation were randomized at birth to receive 5 ppm of iNO if they presented with hypoxemic respiratory failure. Nonhypoxemic infants were studied as the reference group. Blood samples were withdrawn 24 h apart, within the first 4 d of life, to assess malondialdehyde (MDA) concentration as oxidative stress marker and total plasmatic glutathione (GSH), intraerythrocyte GSH peroxidase, and GSH reductase activities as antioxidant defenses. After 24 h, the rise in MDA was blunted in the iNO group compared with controls and was close to the reference infants. Conversely, GSH was more stable in the iNO group, when there was no difference for the GSH peroxidase and GSH reductase activities. On day 28, Oxygen dependence was linked with a higher increase in MDA as was the risk for death, whereas intraventricular hemorrhage was associated with a higher initial drop in GSH. Early low-dose iNO in hypoxemic preterm infants improves oxidative balance and seems to be clinically beneficial up to day 28 of life. PMID- 15746262 TI - Nuclear factor kappaB activation in pulmonary leukocytes from infants with hyaline membrane disease: associations with chorioamnionitis and Ureaplasma urealyticum colonization. AB - Unresolved pulmonary inflammation in hyaline membrane disease (HMD) may be a precursor to the development of chronic lung disease of early infancy. We investigated whether nuclear factor kappaB (NF-kappaB), a transcription factor that regulates the inflammatory process, is activated in pulmonary leukocytes in tracheal aspirates from premature infants with HMD. A total of 172 samples were obtained from 59 infants, two thirds of whom showed NF-kappaB activation in lung neutrophils and macrophages on at least one occasion. Infants who had activated NF-kappaB showed elevated tumor necrosis factor-alpha concentrations in their tracheal aspirates. These infants also required a longer period of mechanical ventilation support. Almost half of the infants with HMD had antenatal exposure to chorioamnionitis on the basis of placental histopathologic examination. These infants had evidence of activated NF-kappaB and elevated cytokines and were more likely to have Ureaplasma urealyticum colonization in their airways. Together, these observations suggest that NF-kappaB activation in pulmonary leukocytes may be involved in the lung inflammatory process in infants with HMD. PMID- 15746264 TI - Self-destructive behavior and the hereafter. PMID- 15746263 TI - Gene expression profiles of peripheral and cord blood mononuclear cells altered by thymic stromal lymphopoietin. AB - Thymic stromal lymphopoietin (TSLP) was reported to induce dendritic cells to produce Th2-attracting chemokines, followed by allergic inflammation through stimulating not only CD4-positive T cells but also CD8-positive T cells. Therefore, in this experiment, GeneChip and hierarchical clustering were applied to screen the molecules in whole immunity triggered by TSLP directly and indirectly using both adult peripheral and cord blood mononuclear cells as well as isolated monocytes. Gene expression profiles screened a variety of molecules that are triggered by TSLP with or without CD40 ligation. In the profile, RNA expressions of indoleamine 2,3-dioxygenase, that is known to induce anergy of T cells and natural killer cells in protecting fetal rejection; many kinds of proteasomes that were reported to trigger cytokine production by inhibiting suppressors of NF-kappaB; and several kinds of chemokines increased, whereas RNA expression of superoxide dismutase 1 decreased, which was unexpected but considered worthy of notice. Expression of chemokines at protein levels and enzymatic activity of indoleamine 2,3-dioxygenase was further confirmed to increase in the presence of TSLP using ELISA and HPLC, respectively. These results suggest that the advent of microarray technology may enable us to screen novel molecular targets to treat TSLP-related allergic inflammation. PMID- 15746265 TI - The function of self-harm behavior in a forensic population. AB - This article examines the functions of self-injurious behavior (SIB) in forensic populations. The cases of four individual offenders who have engaged in SIB are presented. These cases depict the four main functions of self-harm behaviors including (a) suicidal intent, (b) manipulation of the environment, (c) emotion regulation, and (d) a response to psychotic delusions or hallucinations. Assessment and treatment implications are discussed. PMID- 15746266 TI - Correlates and predictors of self-reported suicide attempts among incarcerated youths. AB - The correlates and predictors of suicidal behaviors among 900 young offenders in detention centers in South Australia are investigated. That young offenders are a high-risk population is confirmed, with a quarter of the youths reporting recent suicidal ideation and more than a quarter reporting having made a suicide attempt. Past suicide attempts are significantly predictive of future suicide attempts. Other variables both concurrently and prospectively associated with suicidal behaviors (even after controlling for prior suicide attempt status) are substance use, prolonged dysphoric mood, attention deficit hyperactivity disorder signs, and having a bad temper. These variables not only have predictive value but should also be considered as appropriate targets when intervening to reduce suicide risk. It is suggested that impulsiveness is an underlying common factor. PMID- 15746267 TI - Aggressors against women in prison and in the community: an exploratory study of a differential profile. AB - The current study compares the demographic and psychopathological characteristics of 54 men, who were in prison because of a serious offence of violence against women, and of 42 men, who belonged to a program of community treatment for violence against women in the home. There were no significant differences in demographic variables between the two samples. However, from a psychopathological point of view, psychiatric antecedents and current emotional instability were much more frequent and severe in aggressors within the community. Therefore, two possible differential profiles among the violent men are presented. Implications of these results for further research and clinical practice are commented on. PMID- 15746268 TI - The impact of sex offender residence restrictions: 1,000 feet from danger or one step from absurd? AB - Several states have enacted public policies that prohibit sex offenders who have abused children from living within close proximity to a school, park, day care center, or school bus stop. The purpose of this exploratory study was to describe the impact of residence restrictions on sex offender reintegration and to better understand sex offenders' perceptions of these laws. A survey of 135 sex offenders in Florida was conducted. Most of the molesters who responded to the survey indicated that housing restrictions increased isolation, created financial and emotional stress, and led to decreased stability. Respondents also indicated that they did not perceive residence restrictions as helpful in risk management and, in fact, reported that such restrictions may inadvertently increase triggers for reoffense. Implications for policy and practice are discussed. PMID- 15746270 TI - Naming oneself criminal: gender difference in offenders' identity negotiation. AB - This qualitative research examines gender differences in offenders'ability to negotiate a positive identity once the pejorative labels of criminal, prostitute, drug dealer, and incompetent parents have been imputed onto them. In-depth semi structured focused interviews were conducted with a purposeful information-rich sample of eight male and eight female offenders. Content analysis reveals that males were much more adept than female offenders at juggling with conventional and street norms to justify and/or resist stigmatizing labels in order to construct a favorable identity. Appeal to such personal strengths as know-how, competence, loyalty, and a code of honor allowed male offenders to challenge the boundaries between conventional and delinquent worlds. Concomitantly such an appeal gave rise to a sense of efficacy perception and an optimistic explanatory style. In contrast, even though female offenders were able to justify the labels of drug dealer, prostitute, and thief by appeal to higher loyalties and reject that of insane, all their justifications collapsed when having to negotiate the identity of incompetent mother. Female offenders' negative internal attributions and deprivation of the normative center-motherhood resulted in apathy, anomie, and lack of confidence in their ability to do something worthwhile. Rehabilitation guidelines would build female offenders' personal strengths while redirecting those exhibited by male offenders into lawful enterprises. PMID- 15746269 TI - Prevalence of co-occurring disorders among juveniles committed to detention centers. AB - The Practical Adolescent Diagnostic Interview (PADDI), a structured diagnostic interview designed to gather basic information about substance use disorders, mental health conditions, and related experiences, was used in routine evaluation of all adolescents committed to two juvenile detention centers. Anonymous data from 252 consecutively admitted adolescents were analyzed to assess the prevalence and severity of problem areas and to provide a preliminary exploration of interrelationships among those problems. Results demonstrated that a majority of adolescents manifested multiple problems. Whereas prevalence rates were high across gender, females displayed significantly greater mental health problems and maltreatment histories. PMID- 15746271 TI - Nonviolent offenders' and college students' attachment and social support behaviors: implications for counseling. AB - This study examines the relationships between adult attachment orientations and the ability to seek social support as factors in mediating behavior. Data were collected on non violent offenders (n = 57) and college students (n = 89). The results indicated the non-normative group of nonviolent offenders reported being more securely attached and having more satisfaction with their social supports than the normative group of college students. Fewer college students reported being securely attached than the nonviolent offenders but had a larger number of social supports. In the insecure categories, nonviolent offenders reported being more dismissively attached whereas college students reported being preoccupied in their attachment orientation. The study suggests that attachment and social support-seeking behaviors across different populations may be potential factors in designing counseling services that are used for risk classification and needs assessment. PMID- 15746272 TI - Comment on Mailloux et al.'s (2003) Study "Dosage of treatment to sexual offenders: are we overprescribing?". PMID- 15746274 TI - Computational prediction and experimental verification of novel IdeR binding sites in the upstream sequences of Mycobacterium tuberculosis open reading frames. AB - IdeR (iron-dependent regulator) is a key regulator of virulence factors and iron acquisition systems in Mycobacterium tuberculosis. Despite the wealth of information available on IdeR-regulated genes of M.tuberculosis, there is still an underlying possibility that there are novel genes/pathways that have gone undetected, the identification of which could give new insights into understanding the pathogenesis of M.tuberculosis. We describe an in silico approach employing the positional relative entropy method to identify potential IdeR binding sites in the upstream sequences of all the 3919 ORFs of M.tuberculosis. While many of the predictions made by this approach overlapped with the ones already identified by microarray experiments and binding assays, pointing to the accuracy of our method, a few genes for which there has been no evidence for IdeR regulation were additionally identified. Our results have implications on the iron-dependent regulatory mechanism of M.tuberculosis vis-a vis the activity of urease operon and novel transcription regulators and transporters. PMID- 15746275 TI - Partition-distance via the assignment problem. AB - MOTIVATION: Accuracy testing of various pedigree reconstruction methods requires an efficient algorithm for the calculation of distance between a known partition and its reconstruction. The currently used algorithm of Almudevar and Field takes a prohibitively long time for certain partitions and population sizes. RESULTS: We present an algorithm that very efficiently reduces the partition-distance calculation to the classic assignment problem of weighted bipartite graphs that has known polynomial-time solutions. The performance of the algorithm is tested against the Almudevar and Field partition-distance algorithm to verify the significant improvement in speed. AVAILABILITY: Computer code written in java is available upon request from the first author. PMID- 15746276 TI - Prediction of subcellular localization using sequence-biased recurrent networks. AB - MOTIVATION: Targeting peptides direct nascent proteins to their specific subcellular compartment. Knowledge of targeting signals enables informed drug design and reliable annotation of gene products. However, due to the low similarity of such sequences and the dynamical nature of the sorting process, the computational prediction of subcellular localization of proteins is challenging. RESULTS: We contrast the use of feed forward models as employed by the popular TargetP/SignalP predictors with a sequence-biased recurrent network model. The models are evaluated in terms of performance at the residue level and at the sequence level, and demonstrate that recurrent networks improve the overall prediction performance. Compared to the original results reported for TargetP, an ensemble of the tested models increases the accuracy by 6 and 5% on non-plant and plant data, respectively. AVAILABILITY: The Protein Prowler incorporating the recurrent network predictor described in this paper is available online at http://pprowler.imb.uq.edu.au/ PMID- 15746277 TI - The mutated subsequence problem and locating conserved genes. AB - MOTIVATION: For the purpose of locating conserved genes in a whole genome scale, this paper proposes a new structural optimization problem called the Mutated Subsequence Problem, which gives consideration to possible mutations between two species (in the form of reversals and transpositions) when comparing the genomes. RESULTS: A practical algorithm called mutated subsequence algorithm (MSS) is devised to solve this optimization problem, and it has been evaluated using different pairs of human and mouse chromosomes, and different pairs of virus genomes of Baculoviridae. MSS is found to be effective and efficient; in particular, MSS can reveal >90% of the conserved genes of human and mouse that have been reported in the literature. When compared with existing softwares MUMmer and MaxMinCluster, MSS uncovers 14 and 7% more genes on average, respectively. Furthermore, this paper shows a hybrid approach to integrate MUMmer or MaxMinCluster with MSS, which has better performance and reliability. PMID- 15746278 TI - A periodicity analysis of transmembrane helices. AB - Transmembrane helices and the helical bundles which they form are the major building blocks of membrane proteins. Since helices are characterized by a given periodicity, it is possible to search for patterns of traits which typify one side of the helix and not the other (e.g. amphipathic helices contain a polar and apolar sides). Using Fourier transformation we have analyzed solved membrane protein structures as well as sequences of membrane proteins from the Swiss-Prot database. The traits searched included aromaticity, volume and ionization. While a number of motifs were already recognized in the literature, many were not. One particular example involved helix VII of lactose permease which contains seven aromatic residues on six helical turns. Similarly six glycine residues in four consecutive helical turns were identified as forming a motif in the chloride channel. A tabulation of all the findings is presented as well as a possible rationalization of the function of the motif. PMID- 15746279 TI - Classification of bacterial species from proteomic data using combinatorial approaches incorporating artificial neural networks, cluster analysis and principal components analysis. AB - MOTIVATION: Robust computer algorithms are required to interpret the vast amounts of proteomic data currently being produced and to generate generalized models which are applicable to 'real world' scenarios. One such scenario is the classification of bacterial species. These vary immensely, some remaining remarkably stable whereas others are extremely labile showing rapid mutation and change. Such variation makes clinical diagnosis difficult and pathogens may be easily misidentified. RESULTS: We applied artificial neural networks (Neuroshell 2) in parallel with cluster analysis and principal components analysis to surface enhanced laser desorption/ionization (SELDI)-TOF mass spectrometry data with the aim of accurately identifying the bacterium Neisseria meningitidis from species within this genus and other closely related taxa. A subset of ions were identified that allowed for the consistent identification of species, classifying >97% of a separate validation subset of samples into their respective groups. AVAILABILITY: Neuroshell 2 is commercially available from Ward Systems. PMID- 15746280 TI - Integration of GO annotations in Correspondence Analysis: facilitating the interpretation of microarray data. AB - MOTIVATION: The functional interpretation of microarray datasets still represents a time-consuming and challenging task. Up to now functional categories that are relevant for one or more experimental context(s) have been commonly extracted from a set of regulated genes and presented in long lists. RESULTS: To facilitate interpretation, we integrated Gene Ontology (GO) annotations into Correspondence Analysis to display genes, experimental conditions and gene-annotations in a single plot. The position of the annotations in these plots can be directly used for the functional interpretation of clusters of genes or experimental conditions without the need for comparing long lists of annotations. Correspondence Analysis is not limited in the number of experimental conditions that can be compared simultaneously, allowing an easy identification of characterizing annotations even in complex experimental settings. Due to the rapidly increasing amount of annotation data available, we apply an annotation filter. Hereby the number of displayed annotations can be significantly reduced to a set of descriptive ones, further enhancing the interpretability of the plot. We validated the method on transcription data from Saccharomyces cerevisiae and human pancreatic adenocarcinomas. AVAILABILITY: The M-CHiPS software is accessible for collaborators at http://www.mchips.org PMID- 15746281 TI - Prediction of the phenotypic effects of non-synonymous single nucleotide polymorphisms using structural and evolutionary information. AB - MOTIVATION: There has been great expectation that the knowledge of an individual's genotype will provide a basis for assessing susceptibility to diseases and designing individualized therapy. Non-synonymous single nucleotide polymorphisms (nsSNPs) that lead to an amino acid change in the protein product are of particular interest because they account for nearly half of the known genetic variations related to human inherited diseases. To facilitate the identification of disease-associated nsSNPs from a large number of neutral nsSNPs, it is important to develop computational tools to predict the phenotypic effects of nsSNPs. RESULTS: We prepared a training set based on the variant phenotypic annotation of the Swiss-Prot database and focused our analysis on nsSNPs having homologous 3D structures. Structural environment parameters derived from the 3D homologous structure as well as evolutionary information derived from the multiple sequence alignment were used as predictors. Two machine learning methods, support vector machine and random forest, were trained and evaluated. We compared the performance of our method with that of the SIFT algorithm, which is one of the best predictive methods to date. An unbiased evaluation study shows that for nsSNPs with sufficient evolutionary information (with not <10 homologous sequences), the performance of our method is comparable with the SIFT algorithm, while for nsSNPs with insufficient evolutionary information (<10 homologous sequences), our method outperforms the SIFT algorithm significantly. These findings indicate that incorporating structural information is critical to achieving good prediction accuracy when sufficient evolutionary information is not available. AVAILABILITY: The codes and curated dataset are available at http://compbio.utmem.edu/snp/dataset/ PMID- 15746282 TI - Mega2: data-handling for facilitating genetic linkage and association analyses. AB - Mega2, the manipulation environment for genetic analysis, transparently allows users to process genetic data for family-based or case/control studies accurately and efficiently. In addition to data validation checks, Mega2 provides analysis setup capabilities for a broad choice of commonly used genetic analysis programs, including SimWalk2, ASPEX, GeneHunter, SLINK, SIMULATE, S.A.G.E., SOLAR, Vitesse, Allegro, PREST, PAP, Loki, Merlin and MENDEL. AVAILABILITY: http://watson.hgen.pitt.edu/register/ PMID- 15746283 TI - Prediction of protein interdomain linker regions by a hidden Markov model. AB - MOTIVATION: Our aim was to predict protein interdomain linker regions using sequence alone, without requiring known homology. Identifying linker regions will delineate domain boundaries, and can be used to computationally dissect proteins into domains prior to clustering them into families. We developed a hidden Markov model of linker/non-linker sequence regions using a linker index derived from amino acid propensity. We employed an efficient Bayesian estimation of the model using Markov Chain Monte Carlo, Gibbs sampling in particular, to simulate parameters from the posteriors. Our model recognizes sequence data to be continuous rather than categorical, and generates a probabilistic output. RESULTS: We applied our method to a dataset of protein sequences in which domains and interdomain linkers had been delineated using the Pfam-A database. The prediction results are superior to a simpler method that also uses linker index. PMID- 15746284 TI - Fast tandem mass spectra-based protein identification regardless of the number of spectra or potential modifications examined. AB - MOTIVATION: Comparing tandem mass spectra (MSMS) against a known dataset of protein sequences is a common method for identifying unknown proteins; however, the processing of MSMS by current software often limits certain applications, including comprehensive coverage of post-translational modifications, non specific searches and real-time searches to allow result-dependent instrument control. This problem deserves attention as new mass spectrometers provide the ability for higher throughput and as known protein datasets rapidly grow in size. New software algorithms need to be devised in order to address the performance issues of conventional MSMS protein dataset-based protein identification. METHODS: This paper describes a novel algorithm based on converting a collection of monoisotopic, centroided spectra to a new data structure, named 'peptide finite state machine' (PFSM), which may be used to rapidly search a known dataset of protein sequences, regardless of the number of spectra searched or the number of potential modifications examined. The algorithm is verified using a set of commercially available tryptic digest protein standards analyzed using an ABI 4700 MALDI TOFTOF mass spectrometer, and a free, open source PFSM implementation. It is illustrated that a PFSM can accurately search large collections of spectra against large datasets of protein sequences (e.g. NCBI nr) using a regular desktop PC; however, this paper only details the method for identifying peptide and subsequently protein candidates from a dataset of known protein sequences. The concept of using a PFSM as a peptide pre-screening technique for MSMS-based search engines is validated by using PFSM with Mascot and XTandem. AVAILABILITY: Complete source code, documentation and examples for the reference PFSM implementation are freely available at the Proteome Commons, http://www.proteomecommons.org and source code may be used both commercially and non-commercially as long as the original authors are credited for their work. PMID- 15746285 TI - Non-additivity in protein-DNA binding. AB - MOTIVATION: Localizing protein binding sites within genomic DNA is of considerable importance, but remains difficult for protein families, such as transcription factors, which have loosely defined target sequences. It is generally assumed that protein affinity for DNA involves additive contributions from successive nucleotide pairs within the target sequence. This is not necessarily true, and non-additive effects have already been experimentally demonstrated in a small number of cases. The principal origin of non-additivity involves the so-called indirect component of protein-DNA recognition which is related to the sequence dependence of DNA deformation induced during complex formation. Non-additive effects are difficult to study because they require the identification of many more binding sequences than are normally necessary for describing additive specificity (typically via the construction of weight matrices). RESULTS: In the present work we will use theoretically estimated binding energies as a basis for overcoming this problem. Our approach enables us to study the full combinatorial set of sequences for a variety of DNA-binding proteins, make a detailed analysis of non-additive effects and exploit this information to improve binding site predictions using either weight matrices or support vector machines. The results underline the fact that, even in the presence of significant deformation, non-additive effects may involve only a limited number of dinucleotide steps. This information helps to reduce the number of binding sites which need to be identified for successful predictions and to avoid problems of over-fitting. AVAILABILITY: The SVM software is available upon request from the authors. PMID- 15746286 TI - CoC: a database of universally conserved residues in protein folds. AB - The conservatism of conservatism (CoC) database presents statistically analyzed information about the conservation of residue positions in folds across protein families. AVAILABILITY: On the web at http://kulibin.mit.edu/coc/ PMID- 15746287 TI - ADAPT: a database of affymetrix probesets and transcripts. AB - ADAPT is an online database providing comprehensive mappings between Affymetrix probes and RefSeq and Ensembl transcripts. ADAPT was designed to help interpret microarray experiments by providing a means to explore the many-to-many relationships that exist between probes, probesets, transcripts and genes. AVAILABILITY: ADAPT can be queried via the web at http://bioinformatics.picr.man.ac.uk/adapt PMID- 15746288 TI - ArrayCyGHt: a web application for analysis and visualization of array-CGH data. AB - ArrayCyGHt is a web-based application tool for analysis and visualization of microarray-comparative genomic hybridization (array-CGH) data. Full process of array-CGH data analysis, from normalization of raw data to the final visualization of copy number gain or loss, can be straightforwardly achieved on this arrayCyGHt system without the use of any further software. ArrayCyGHt, therefore, provides an easy and fast tool for the analysis of copy number aberrations in any kinds of data format. AVAILABILITY: ArrayCyGHt can be accessed at http://genomics.catholic.ac.kr/arrayCGH/ PMID- 15746289 TI - WebAllergen: a web server for predicting allergenic proteins. AB - WebAllergen is a web server that predicts the potential allergenicity of proteins. The query protein will be compared against a set of prebuilt allergenic motifs that have been obtained from 664 known allergen proteins. The query will also be compared with known allergens that do not have detectable allergenic motifs. Moreover, users are allowed to upload their own allergens as alternative training sequences on which a new set of allergenic motifs will be built. The query sequences can also be compared with these motifs. AVAILABILITY: http://weballergen.bii.a-star.edu.sg/ PMID- 15746290 TI - Gene-Expression Omnibus integration and clustering tools in SeqExpress. AB - SeqExpress, a gene-expression analysis suite, has been extended to offer a number of cluster generation, refinement and visualization techniques. The cluster generation methods have been specialized to deal with aspects of the sparseness and extreme values that occur within microarray data. The results of such cluster analysis can then be refined using either: a functional enrichment based procedure, which examines each cluster to see if it possesses an unusually high or low concentration of ontology terms; or by using Expectation-Maximization to find a mixture of model based distributions within the datasets. Visualizations are provided both to explore and compare the results of the cluster generation algorithms. In addition, a tool has been developed which integrates SeqExpress with the Gene-Expression Omnibus repository. The tool provides seamless access to the large number of experimental results in the repository, so that they can be visualized and analysed locally using SeqExpress. AVAILABILITY: SeqExpress is available as a 6 MB download from http://www.seqexpress.com and runs under Windows. A server-based version is available and is required for the GEO integration. SeqExpress is not affiliated with any academic institution, funding body or commercial organization and is free to use by all. PMID- 15746291 TI - SNP discovery using advanced algorithms and neural networks. AB - Forage is an application which uses two neural networks for detecting single nucleotide polymorphisms (SNPs). Potential SNP candidates are identified in multiple alignments. Each candidate is then represented by a vector of features, which is classified as SNP or monomorphic by the networks. A validated dataset of SNPs was constructed from experimentally verified SNP data and used for network training and method evalutation. AVAILABILITY: The package is available at biobase.biotech.kth.se/forage/ PMID- 15746292 TI - Multiple flexible structure alignment using partial order graphs. AB - MOTIVATION: Existing comparisons of protein structures are not able to describe structural divergence and flexibility in the structures being compared because they focus on identifying a common invariant core and ignore parts of the structures outside this core. Understanding the structural divergence and flexibility is critical for studying the evolution of functions and specificities of proteins. RESULTS: A new method of multiple protein structure alignment, POSA (Partial Order Structure Alignment), was developed using a partial order graph representation of multiple alignments. POSA has two unique features: (1) identifies and classifies regions that are conserved only in a subset of input structures and (2) allows internal rearrangements in protein structures. POSA outperforms other programs in the cases where structural flexibilities exist and provides new insights by visualizing the mosaic nature of multiple structural alignments. POSA is an ideal tool for studying the variation of protein structures within diverse structural families. AVAILABILITY: POSA is freely available for academic users on a Web server at http://fatcat.burnham.org/POSA PMID- 15746293 TI - Contribution of prostaglandins to the dilation that follows isometric forearm contraction in human subjects: effects of aspirin and hyperoxia. AB - In 11 healthy volunteers, we evaluated, in a double-blind crossover study, whether the vasodilation that follows isometric contraction is mediated by prostaglandins (PGs) and/or is O2 dependent. Subjects performed isometric handgrip for 2 min at 60% maximal voluntary contraction (MVC), after pretreatment with placebo or aspirin (600 mg orally), when breathing air or 40% O2. Forearm blood flow was measured in the dominant forearm by venous occlusion plethysmography. Arterial blood pressure was also recorded, allowing calculation of forearm vascular conductance (FVC; forearm blood flow/arterial blood pressure). During air breathing, aspirin significantly reduced the increase in FVC that followed contraction at 60% MVC: from a baseline of 0.09 +/- 0.011 [mean +/- SE, conductance units (CU)], the peak value was reduced from 0.24 +/- 0.03 to 0.14 +/- 0.01 CU. Breathing 40% O2 similarly reduced the increase in FVC relative to that evoked when breathing air; the peak value was 0.24 +/- 0.03 vs. 0.15 +/- 0.02 CU. However, after aspirin, breathing 40% O2 had no further effect on the contraction-evoked increase in FVC (the peak value was 0.15 +/- 0.02 vs. 0.16 +/- 0.02 CU). Thus the present study indicates that prostaglandins make a substantial contribution to the peak of the vasodilation that follows isometric contraction of forearm muscles at 60% MVC. Given that hyperoxia similarly reduced the vasodilation and attenuated the effect of aspirin, we propose that the stimulus for prostaglandin synthesis and release is hypoxia of the endothelium. PMID- 15746294 TI - Power training is more effective than strength training for maintaining bone mineral density in postmenopausal women. AB - Physical exercise has a favorable impact on bones, but optimum training strategies are still under discussion. In this study, we compared the effect of slow and fast resistance exercises on various osteodensitometric parameters. Fifty-three postmenopausal women were randomly assigned to a strength training (ST) or a power training group (PT). Both groups carried out a progressive resistance training, a gymnastics session, and a home training over a period of 12 mo. During the resistance training, the ST group used slow and the PT group fast movements; otherwise there were no training differences. All subjects were supplemented with Ca and vitamin D. At baseline and after 12 mo, bone mineral density (BMD) was measured at the lumbar spine, proximal femur, and distal forearm by dual-energy X-ray absorptiometry. We also measured anthropometric data and maximum static strength. Frequency and grade of pain were assessed by questionnaire. After 12 mo, significant between-group differences were observed for BMD at the lumbar spine (P < 0.05) and the total hip (P < 0.05). Whereas the PT group maintained BMD at the spine (+0.7 +/- 2.1%, not significant) and the total hip (0.0 +/- 1.7%, not significant), the ST group lost significantly at both sites (spine: -0.9 +/- 1.9%; P < 0.05; total hip: -1.2 +/- 1.5%; P < 0.01). No significant between-group differences were observed for anthropometric data, maximum strength, BMD of the forearm, or frequency and grade of pain. These findings suggest that power training is more effective than strength training in reducing bone loss in postmenopausal women. PMID- 15746295 TI - Gene expression of myogenic factors and phenotype-specific markers in electrically stimulated muscle of paraplegics. AB - The transcription factors myogenin and MyoD have been suggested to be involved in maintaining slow and fast muscle-fiber phenotypes, respectively, in rodents. Whether this is also the case in human muscle is unknown. To test this, 4 wk of chronic, low-frequency electrical stimulation training of the tibialis anterior muscle of paraplegic subjects were used to evoke a fast-to-slow transformation in muscle phenotype. It was hypothesized that this would result from an upregulation of myogenin and a downregulation of MyoD. The training evoked the expected mRNA increase for slow fiber-specific markers myosin heavy chain I and 3-hydroxyacyl CoA dehydrogenase A, whereas an mRNA decrease was seen for fast fiber-specific markers myosin heavy chain IIx and glycerol phosphate dehydrogenase. Although the slow fiber-specific markers citrate synthase and muscle fatty acid binding protein did not display a significant increase in mRNA, they did tend to increase. As hypothesized, myogenin mRNA was upregulated. However, contrary to the hypothesis, MyoD mRNA also increased, although later than myogenin. The mRNA levels of the other myogenic regulatory factor family members, myogenic factor 5 and myogenic regulatory factor 4, and the myocyte enhancer factor (MEF) family members, MEF-2A and MEF-2C, did not change. The results indicate that myogenin is indeed involved in the regulation of the slow oxidative phenotype in human skeletal muscle fibers, whereas MyoD appears to have a more complex regulatory function. PMID- 15746296 TI - Plasma from conscious hypoxic rats stimulates leukocyte-endothelial interactions in normoxic cremaster venules. AB - Systemic hypoxia results in rapid increases in leukocyte-endothelial adherence (LEA) and emigration, vascular permeability, and mast cell activation in several microcirculations. Observations in cremaster muscle suggest that this response is initiated by a mediator released from a distant site (Dix R, Orth T, Allen JA, Wood JG, and Gonzalez NC. J Appl Physiol 95: 2495-2502, 2003). The present experiments in rat cremaster muscle tested the hypothesis that, if a circulating mediator triggers hypoxia-induced inflammation, then plasma from hypoxic rats should elicit LEA in normoxic cremaster venules. Plasma from conscious donor rats breathing 10% O2-90% N2 for 5 min was applied topically to the cremaster of normoxic anesthetized rats. In this and all other groups described below, the donor plasma had attained normoxic PO2 when applied to the cremaster. LEA (leukocytes/100-microm venule) increased from 2.7 +/- 0.8 to 12.3 +/- 2.4, and venular shear rate and arteriolar diameter decreased to 79 +/- 9% (P < 0.05, n = 6) and 77 +/- 5% of control (P < 0.05, n = 5), respectively, 10 min after application of plasma from hypoxic donors. The decrease in venular shear rate was exclusively due to a reduction of venular blood flow, secondary to the upstream arteriolar vasoconstriction. Plasma from normoxic donors had no effects. Plasma from blood equilibrated in vitro for 5 min with 5% CO2-95% N2 did not alter LEA or shear rate of normoxic cremasters, suggesting that the putative mediator does not originate in blood cells. The effects of plasma from hypoxic rats persisted when the donors were pretreated with the mast cell stabilizer cromolyn, which prevents hypoxia-induced LEA. This suggests that the effects of hypoxic plasma are not due to inflammatory mediators released by adherent leukocytes in the donor rat. There was a positive correlation between LEA and mast cell degranulation observed histologically. These results support the idea that systemic hypoxia produces the release of a substance transported by the circulation that initiates the microvascular inflammation. PMID- 15746297 TI - Alpha-adrenoreceptor blockade with phenoxybenzamine does not affect the ability of the nose to condition air. AB - The primary function of the nose is to warm and humidify air. We have previously shown that raising nasal mucosal temperature by immersing feet in warm water increases the amount of water evaporated by the nose as air passes through it (nasal conditioning capacity; Abbott D, Baroody F, Naureckas E, and Naclerio R. Am J Rhinol 15: 41-45, 2001). To investigate further the effect of nasal mucosal temperature on nasal conditioning capacity, we raised the temperature through alpha-adrenoreceptor blockade by intranasally administering phenoxybenzamine. We hypothesized that blocking alpha-adrenoreceptors during inhalation of cold, dry air would lead to an increase in nasal blood flow, surface temperature, and nasal conditioning capacity, as measured by the water gradient. After appropriate pilot studies, we performed a double-blind, placebo-controlled, two-way crossover study in nine nonatopic, healthy subjects by studying the effect of treatment with intranasal phenoxybenzamine. Nasal mucosal temperature increased significantly after administration of phenoxybenzamine and was associated with a significantly smaller net decrease in nasal mucosal temperature after exposure to cold, dry air (P < 0.05). However, there were no significant differences in nasal conditioning capacity between treatments (P > 0.05). Phenoxybenzamine decreased the symptom of rhinorrhea after exposure to cold, dry air (P < 0.05), but congestion was not different between individuals given phenoxybenzamine and placebo (P > 0.05). Our data demonstrate that phenoxybenzamine, despite raising mucosal temperature and not affecting nasal volume, did not affect the ability of the nose to warm and humidify air. PMID- 15746298 TI - Force recovery after activated shortening in whole skeletal muscle: transient and steady-state aspects of force depression. AB - The depression of isometric force after active shortening is a well-accepted characteristic of skeletal muscle, yet its mechanisms remain unknown. Although traditionally analyzed at steady state, transient phenomena caused, at least in part, by cross-bridge kinetics may provide novel insight into the mechanisms associated with force depression (FD). To identify the transient aspects of FD and its relation to shortening speed, shortening amplitude, and muscle mechanical work, in situ experiments were conducted in soleus muscle-tendon units of anesthetized cats. The period immediately after shortening, in which force recovers toward steady state, was fit by using an exponential recovery function (R2 > 0.99). Statistical analyses revealed that steady-state FD (FD(ss)) increased with shortening amplitude and mechanical work. This FD(ss) increase was always accompanied by a significant decrease in force recovery rate. Furthermore, a significant reduction in stiffness was observed after all activated shortenings, presumably because of a reduced proportion of attached cross bridges. These results were interpreted with respect to the two most prominent proposed mechanisms of force depression: sarcomere length nonuniformity theory (7, 32) and a stress-induced inhibition of cross-bridge binding in the newly formed actin-myosin overlap zone (14, 28). We hypothesized that the latter could describe both steady-state and transient aspects of FD using a single scalar variable, the mechanical work done during shortening. As either excursion (overlap) or force (stress) is increased, mechanical work increases, and cross bridge attachment would become more inhibited, as supported by this study in which an increase in mechanical work resulted in a slower recovery to a more depressed steady-state force. PMID- 15746300 TI - The metabolic syndrome: requiescat in pace. AB - Values for insulin-mediated glucose disposal vary continuously throughout a population of apparently healthy individuals, with at least a sixfold variation between the most insulin sensitive and most insulin resistant of these individuals. The more insulin resistant a person, the more insulin must be secreted to prevent decompensation of glucose tolerance. Insulin resistance is not a disease, but a description of a physiologic state, and approximately one third of an apparently healthy population is sufficiently insulin resistant to be at increased risk to develop a cluster of abnormalities and related clinical syndromes. The primary value of the concept of insulin resistance is that it provides a conceptual framework with which to place a substantial number of apparently unrelated biological events into a pathophysiological construct. In contrast, the metabolic syndrome was introduced as a diagnostic category to identify individuals that satisfy three of five relatively arbitrarily chosen criteria to initiate lifestyle changes with the goal of decreasing risk of cardiovascular disease. Consequently, the value of the notion of the metabolic syndrome must be considered not in pathophysiologic terms, but as a pragmatic approach to obtain a better clinical outcome. In this review, an effort is made to critically evaluate the concept of the metabolic syndrome, the criteria chosen to identify individuals with the syndrome, and the clinical utility of making, or not making, a diagnosis of the metabolic syndrome. PMID- 15746299 TI - Changes in myosin heavy chain mRNA and protein expression in human skeletal muscle with age and endurance exercise training. AB - Aging is associated with reduced muscle strength and atrophy of type II muscle fibers. Muscle fiber type and contractile function are primarily determined by myosin heavy chain (MHC) isoforms. There are few data available on the effects of aging on MHC isoform expression in humans. In the present study, we tested the hypothesis that MHC isoform protein composition and mRNA abundance would favor a fast-to-slow isoform shift with aging and in response to endurance exercise training. Muscle biopsies were obtained from previously sedentary, healthy men and women, aged 21-87 yr before (n = 77) and after (n = 65) 16 wk of bicycle training (up to 45 min at 80% peak heart rate, 3-4 days/wk). At baseline, MHC I mRNA was unchanged with age, whereas IIa and IIx declined by 14 and 10% per decade, respectively (P < 0.001). MHC IIa and IIx protein declined by 3 and 1% per decade with a reciprocal increase in MHC I (P < 0.05). After training, MHC I and IIa mRNA increased by 61 and 99%, respectively, and IIx decreased by 50% (all P < 0.001). The increase in MHC I mRNA was positively associated with age, whereas the changes in MHC IIa and IIx mRNA were similar across age. MHC I protein increased by 6% and was positively related to age, whereas IIx decreased by 5% and was inversely related to age. These results suggest that the altered expression of MHC isoforms with aging is transcriptionally regulated. In response to endurance exercise, regulation of MHC isoform transcripts remains robust in older muscle, but this did not result in corresponding changes in MHC protein expression. PMID- 15746301 TI - Quantification of mycophenolic acid and glucuronide metabolite in human serum by HPLC-tandem mass spectrometry. AB - BACKGROUND: The potent immunosuppressant mycophenolic acid (MPA) is metabolized to an inactive glucuronide (MPAG). The extent of metabolism varies among individuals, and the MPAG formed can be hydrolyzed to MPA and can displace MPA from serum albumin, creating a potential need to monitor both MPA and MPAG. METHODS: After addition of the carboxybutoxy ether of MPA (MPAC) as internal standard, MPA and MPAG were isolated from serum by acidification followed by solid-phase extraction. Gradient chromatographic separation was performed on a Waters Atlantis reversed-phase liquid chromatography (HPLC) column, and the compounds were quantified by electrospray ionization tandem mass spectrometry (MS/MS) in the multiple-reaction monitoring mode. Results obtained by HPLC-MS/MS were compared with an HPLC assay using ultraviolet detection (HPLC-UV) performed at a reference laboratory. RESULTS: MPAG, MPA, and MPAC were fully separated during a 7.0-min run time. Precision at both low and high concentrations of MPA ad MPAG met the suggested method validation criteria from a consensus panel report on MPA. The extraction efficiencies were 99% for MPA and MPAG. The assay was linear to 16 mg/L for MPA and 200 mg/L for MPAG. Limits of quantification were 0.1 mg/L for MPA and 1 mg/L for MPAG. Regression analysis gave the following results: HPLC-MS/MS = 1.03(HPLC-UV) - 0.03 mg/L (R2 = 0.982) for MPA; and HPLC MS/MS = 0.93(HPLC-UV) + 0.89 mg/L (R2 = 0.967) for MPAG. CONCLUSION: This HPLC MS/MS assay can be used to reproducibly quantify MPA and MPAG across a large analytical range in serum from organ transplant patients. PMID- 15746302 TI - Technology for automated, rapid, and quantitative PCR or reverse transcription PCR clinical testing. AB - BACKGROUND: PCR-based assays can improve clinical care, but they remain technically demanding and labor-intensive. We describe a new instrument, the GeneXpert, that performs automated nucleic acid isolation, reverse transcription, and fluorescence-based quantitative PCR in approximately 35 min. METHODS: Yield and integrity of RNA isolated on the GeneXpert were compared with Qiagen-based extraction for parallel samples (5-microm frozen tissue sections). The reproducibility of automated RNA isolation, reverse transcription, and quantitative PCR was determined by replicate (n = 10) analysis of 10 tissues, using duplex (target and endogenous control) reverse transcription-PCR reactions for two gene combinations. The GeneXpert was then used to perform rapid analysis of lymph nodes from melanoma, breast cancer, and lung cancer patients and analysis of melanoma metastatic to the lung, primary lung adenocarcinoma, and healthy lung tissue. RESULTS: On the GeneXpert, RNA was recovered in slightly over 6 min, and the yield was approximately 70% of that from parallel Qiagen reactions. The RNA integrity was comparable to that of Qiagen-isolated RNA as determined by gel electrophoresis. For the melanoma samples, the 95% prediction interval for the deltaCt for a new measurement was +/-1.54 cycles, and for breast cancer samples, the interval for a newly observed deltaCt was +/-1.40 cycles. GeneXpert assays successfully detected the presence of metastatic melanoma, breast cancer, and lung cancer in lymph nodes and also differentiated among metastatic melanoma, lung adenocarcinoma, and healthy lung. CONCLUSIONS: RNA yield and integrity on the GeneXpert are comparable to benchtop methods. Reproducibility of the GeneXpert data is similar to that seen with manual methods in our hands but may need improvement for some applications. The GeneXpert can perform RNA isolation, reverse transcription, and quantitative PCR in approximately 35 min and could therefore be used for intraoperative testing when applicable. PMID- 15746303 TI - Brain stem melanocortinergic modulation of meal size and identification of hypothalamic POMC projections. AB - Metabolic, cognitive, and environmental factors processed in the forebrain modulate food intake by changing the potency of direct controls of meal ingestion in the brain stem. Here, we behaviorally and anatomically test the role of the hypothalamic proopiomelanocortin (POMC) system in mediating some of these descending, indirect controls. Melanotan II (MTII), a stable melanocortin 4 receptor (MC4R) and melanocortin 3 receptor (MC3R) agonist injected into the fourth ventricle near the dorsal vagal complex, potently inhibited 14-h food intake by decreasing meal size but not meal frequency; SHU9119, an antagonist, increased food intake by selectively increasing meal size. Furthermore, MTII injected into the fourth ventricle increased and SHU9119 tended to decrease heart rate and body temperature measured telemetrically in freely moving rats. Numerous alpha-melanocyte-stimulating hormone-immunoreactive axons were in close anatomical apposition to nucleus tractus solitarius neurons showing c-Fos in response to gastric distension, expressing neurochemical phenotypes implicated in ingestive control, and projecting to brown adipose tissue. In retrograde tracing experiments, a small percentage of arcuate nucleus POMC neurons was found to project to the dorsal vagal complex. Thus melanocortin signaling in the brain stem is sufficient to alter food intake via changing the potency of satiety signals and to alter sympathetic outflow. Although the anatomical findings support the involvement of hypothalamomedullary POMC projections in mediating part of the descending, indirect signal, they do not rule out involvement of POMC neurons in the nucleus tractus solitarius in mediating part of the direct signal. PMID- 15746304 TI - Periprandial changes in growth hormone release in goldfish: role of somatostatin, ghrelin, and gastrin-releasing peptide. AB - In goldfish, growth hormone (GH) transiently rises 30 min after meals, returning to baseline at 1 h postmeal. Somatostatin (SRIF) is the major inhibitor of GH release. Three cDNAs encoding pre-pro-SRIF (PSS) have been previously cloned from goldfish brain: PSS-I, which encodes SRIF-14; PSS-II, which is potentially processed into gSRIF-28 that has [Glu(1),Tyr(7)(,)Gly(10)]SRIF-14 at the COOH terminus; and PSS-III, which encodes [Pro(2)]SRIF-14 at its COOH terminus. In goldfish, bombesin (BBS), mimicking the endogenous gastrin-releasing peptide (GRP), acutely suppresses food intake and also stimulates GH release. Ghrelin was recently characterized in goldfish as a GH secretagogue and an orexigen. In this paper, we studied the changes in SRIF mRNA levels during feeding and analyzed the influences of BBS and ghrelin peptides on forebrain PSS expression. The results showed a 60% reduction in PSS-II mRNA after meals, but no changes in the expression of PSS-I and PSS-III were found. Intraperitoneal injections of 100 ng/g body wt of BBS increased GH secretion and decreased PSS-I and PSS-II gene expression. Intraperitoneal injection of goldfish ghrelin (100 ng/g body wt) transiently increased the serum GH levels and increased PSS-I, while decreasing PSS-II mRNA levels. Ghrelin (50 ng/g body wt) blocked the effects of BBS (100 ng/g body wt) on PSS-I but not on PSS-II expression. Coadministration of BBS and ghrelin decreased only the PSS-II gene expression. We conclude that the interactions between BBS/GRP and ghrelin can account for the postprandial variations in serum GH levels and the forebrain expression of PSS-II. Furthermore, we demonstrate that intraperitoneal administration of BBS reduces the ghrelin expression levels in the gut. Thus the inhibition of production of ghrelin in the gut may contribute to the satiety effects of BBS/GRP peptides. PMID- 15746305 TI - Nutritional and hormonal control of lipolysis in isolated gilthead seabream (Sparus aurata) adipocytes. AB - We examined the effects of diet composition and fasting on lipolysis of freshly isolated adipocytes from gilthead seabream (Sparus aurata). We also analyzed the effects of insulin, glucagon, and growth hormone (GH) in adipocytes isolated from fish fed with different diets. Basal lipolysis, measured as glycerol release, increased proportionally with cell concentration and time of incubation, which validates the suitability of these cell preparations for the study of hormonal regulation of this metabolic process. Gilthead seabream were fed two different diets, FM (100% of fish meal) and PP (100% of plant protein supplied by plant sources) for 6 wk. After this period, each diet group was divided into two groups: fed and fasted (for 11 days). Lipolysis was significantly higher in adipocytes from PP-fed fish than in adipocytes from FM-fed fish. Fasting provoked a significant increase in the lipolytic rate, about threefold in isolated adipocytes regardless of nutritional history. Hormone effects were similar in the different groups: glucagon increased the lipolytic rate, whereas insulin had almost no effect. GH was clearly lipolytic, although the relative increase in glycerol over control was lower in isolated adipocytes from fasted fish compared with fed fish. Together, we demonstrate for the first time that lipolysis, measured in isolated seabream adipocytes, is affected by the nutritional state of the fish. Furthermore, our data suggest that glucagon and especially GH play a major role in the control of adipocyte lipolysis. PMID- 15746306 TI - Influence of size at birth on the endocrine profiles and expression of uncoupling proteins in subcutaneous adipose tissue, lung, and muscle of neonatal pigs. AB - Epidemiological studies suggest that infants of low birth weight show poor neonatal growth and increased susceptibility to adult diseases such as diabetes and lung disease. Uncoupling protein 2 and 3 (UCP2 and UCP3) have been implicated in the development of such diseases; pigs provide an ideal model to examine the influence of birth weight due to the natural variance in piglet weight within a litter. This study examined whether birth weight influences the expression of UCP2 and UCP3 in adipose tissue, skeletal muscle, and lung. Piglets from 11 litters were ranked according to birth weight and three from each litter assigned to small (SFD), normal (NFD), or large for dates (LFD) groups. Blood samples and morphometric measurements were taken over the first 14 days of life, and tissue samples were taken on day 7 or 14. Plasma hormone and metabolite concentrations and the expression of UCP2 and UCP3 mRNA in adipose tissue, skeletal muscle, and lung were measured. UCP2 and UCP3 expression in adipose tissue was lower in the SFD compared with the LFD group on day 7. UCP3 expression in skeletal muscle was higher than that of adipose tissue. Lung UCP2 and skeletal muscle UCP3 mRNA expression were unaffected by size at birth. Regression analysis indicated that UCP3 expression was differentially associated with IGF-1, leptin, and insulin. In conclusion, low birth weight is associated with tissue-specific effects on UCP expression. It remains to be established whether these subsequently contribute to pathological conditions such as diabetes. PMID- 15746307 TI - Nonlinearities and asymmetries of the human cardiovagal baroreflex. AB - To determine whether an approach such as the modified Oxford technique can consistently produce data that reveal the nonlinear nature of the cardiovagal baroreflex and to ascertain whether the model parameters provide unique insight into baroreflex function, we retrospectively examined 91 baroreflex trials (38 subjects, 27 men and 11 women, ages 22-72 yr). The modified Oxford technique (bolus sodium nitroprusside followed by bolus phenylephrine) was used to perturb blood pressure, and the resulting systolic blood pressure-R-R interval responses were plotted and modeled using a linear, a four-parameter symmetric, and a five parameter asymmetric model. Several issues, such as the effect of data averaging, various approaches to gain estimation, and the predictive value of model parameters, were examined during reflex modeling. Sigmoid models accounted for a greater amount of the variance than did the linear model: linear r2=0.81+/-0.01, four-parameter r2=0.90+/-0.08, and five-parameter r2=0.90+/-0.08 (P<0.05, linear vs. sigmoid models). Data averaging did not affect model fits. Although the four gain estimates (linear remodel, 1st derivative, peak, and set point) were statistically related, the set point gain was significantly lower than other estimates (P<0.05). Subgroup comparisons between young and older healthy subjects revealed differences in all indexes of cardiovagal baroreflex gain, as well as R R interval operating range and curvature parameters. In conclusion, the modified Oxford technique consistently reveals the nonlinear nature of the human cardiovagal baroreflex. Moreover, of the parameters produced by the symmetric sigmoid model, only the response range provides unique information beyond that of reflex gain. PMID- 15746308 TI - Differential degradation of nonylphenol isomers by Sphingomonas xenophaga Bayram. AB - Sphingomonas xenophaga Bayram, isolated from the activated sludge of a municipal wastewater treatment plant, was able to utilize 4-(1-ethyl-1,4 dimethylpentyl)phenol, one of the main isomers of technical nonylphenol mixtures, as a sole carbon and energy source. The isolate degraded 1 mg of 4-(1-ethyl-1,4 dimethylpentyl)phenol/ml in minimal medium within 1 week. Growth experiments with five nonylphenol isomers showed that the three isomers with quaternary benzylic carbon atoms [(1,1,2,4-tetramethylpentyl)phenol, 4-(1-ethyl-1,4 dimethylpentyl)phenol, and 4-(1,1-dimethylheptyl)phenol] served as growth substrates, whereas the isomers containing one or two hydrogen atoms in the benzylic position [4-(1-methyloctyl)phenol and 4-n-nonylphenol] did not. However, when the isomers were incubated as a mixture, all were degraded to a certain degree. Differential degradation was clearly evident, as isomers with more highly branched alkyl side chains were degraded much faster than the others. Furthermore, the C9 alcohols 2,3,5-trimethylhexan-2-ol, 3,6-dimethylheptan-3-ol, and 2-methyloctan-2-ol, derived from the three nonylphenol isomers with quaternary benzylic carbon atoms, were detected in the culture fluid by gas chromatography-mass spectrometry, but no analogous metabolites could be found originating from 4-(1-methyloctyl)phenol and 4-n-nonylphenol. We propose that 4 (1-methyloctyl)phenol and 4-n-nonylphenol were cometabolically transformed in the growth experiments with the mixture but that, unlike the other isomers, they did not participate in the reactions leading to the detachment of the alkyl moiety. This hypothesis was corroborated by the observed accumulation in the culture fluid of an as yet unidentified metabolite derived from 4-(1-methyloctyl)phenol. PMID- 15746309 TI - Immobilization of Escherichia coli cells by use of the antimicrobial peptide cecropin P1. AB - An immobilization scheme for bacterial cells is described, in which the antimicrobial peptide cecropin P1 was used to trap Escherichia coli K-12 and O157:H7 cells on microtiter plate well surfaces. Cecropin P1 was covalently attached to the well surfaces, and E. coli cells were allowed to bind to the peptide-coated surface. The immobilized cells were detected colorimetrically with an anti-E. coli antibody-horseradish peroxidase conjugate. Binding curves were obtained in which the signal intensities were dependent upon the cell concentration and upon the amount of peptide attached to the well surface. After normalization for the amount of peptide coupled to the surface and the relative binding affinity of the antibody for each strain, the binding data were compared, which indicated that there was a strong preference for E. coli O157:H7 over E. coli K-12. The cells could be immobilized reproducibly at pH values ranging from 5 to 10 and at ionic strengths up to 0.50 M. PMID- 15746310 TI - Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates and for relief of PCR inhibitors. AB - Extraction of high-quality DNA is a key step in PCR detection of Cryptosporidium and other pathogens in environmental samples. Currently, Cryptosporidium oocysts in water samples have to be purified from water concentrates before DNA is extracted. This study compared the effectiveness of six DNA extraction methods (DNA extraction with the QIAamp DNA minikit after oocyst purification with immunomagnetic separation and direct DNA extraction methods using the FastDNA SPIN kit for soil, QIAamp DNA stool minikit, UltraClean soil kit, or QIAamp DNA minikit and the traditional phenol-chloroform technique) for the detection of Cryptosporidium with oocyst-seeded samples, DNA-spiked samples, and field water samples. The study also evaluated the effects of different PCR facilitators (nonacetylated bovine serum albumin, the T4 gene 32 protein, and polyvinylpyrrolidone) and treatments (the use of GeneReleaser or ultrafiltration) for the relief from or removal of inhibitors of PCR amplification. The results of seeding and spiking studies showed that PCR inhibitors were presented in all DNA solutions extracted by the six methods. However, the effect of PCR inhibitors could be relieved significantly by the addition of 400 ng of bovine serum albumin/mul or 25 ng of T4 gene 32 protein/mul to the PCR mixture. With the inclusion of bovine serum albumin in the PCR mixture, DNA extracted with the FastDNA SPIN kit for soil without oocyst isolation resulted in PCR performance similar to that produced by the QIAamp DNA minikit after oocysts were purified by immunomagnetic separation. PMID- 15746311 TI - Validation of a flour-free model dough system for throughput studies of baker's yeast. AB - Evaluation of gene expression in baker's yeast requires the extraction and collection of pure samples of RNA. However, in bread dough this task is difficult due to the complex composition of the system. We found that a liquid model system can be used to analyze the transcriptional response of industrial strains in dough with a high sugar content. The production levels of CO2 and glycerol by two commercial strains in liquid and flour-based doughs were correlated. We extracted total RNA from both a liquid and a flour-based dough. We used Northern blotting to analyze mRNA levels of three stress marker genes, HSP26, GPD1, and ENA1, and 10 genes in different metabolic subcategories. All 13 genes had the same transcriptional profile in both systems. Hence, the model appears to effectively mimic the environment encountered by baker's yeast in high-sugar dough. The liquid dough can be used to help understand the connections between technological traits and biological functions and to facilitate studies of gene expression under commercially important, but experimentally intractable, conditions. PMID- 15746312 TI - Efficient DNA fingerprinting of Clostridium botulinum types A, B, E, and F by amplified fragment length polymorphism analysis. AB - Amplified fragment length polymorphism (AFLP) analysis was applied to characterize 33 group I and 37 group II Clostridium botulinum strains. Four restriction enzyme and 30 primer combinations were screened to tailor the AFLP technique for optimal characterization of C. botulinum. The enzyme combination HindIII and HpyCH4IV, with primers having one selective nucleotide apiece (Hind-C and Hpy-A), was selected. AFLP clearly differentiated between C. botulinum groups I and II; group-specific clusters showed <10% similarity between proteolytic and nonproteolytic C. botulinum strains. In addition, group-specific fragments were detected in both groups. All strains studied were typeable by AFLP, and a total of 42 AFLP types were identified. Extensive diversity was observed among strains of C. botulinum type E, whereas group I had lower genetic biodiversity. These results indicate that AFLP is a fast, highly discriminating, and reproducible DNA fingerprinting method with excellent typeability, which, in addition to its suitability for typing at strain level, can be used for C. botulinum group identification. PMID- 15746313 TI - Induction of Shiga toxin-converting prophage in Escherichia coli by high hydrostatic pressure. AB - Since high hydrostatic pressure is becoming increasingly important in modern food preservation, its potential effects on microorganisms need to be thoroughly investigated. In this context, mild pressures (<200 MPa) have recently been shown to induce an SOS response in Escherichia coli MG1655. Due to this response, we observed a RecA- and LexA-dependent induction of lambda prophage upon treating E. coli lysogens with sublethal pressures. In this report, we extend this observation to lambdoid Shiga toxin (Stx)-converting bacteriophages in MG1655, which constitute an important virulence trait in Stx-producing E. coli strains (STEC). The window of pressures capable of inducing Stx phages correlated well with the window of bacterial survival. When pressure treatments were conducted in whole milk, which is known to promote bacterial survival, Stx phage induction could be observed at up to 250 MPa in E. coli MG1655 and at up to 300 MPa in a pressure-resistant mutant of this strain. In addition, we found that the intrinsic pressure resistance of two types of Stx phages was very different, with one type surviving relatively well treatments of up to 400 MPa for 15 min at 20 degrees C. Interestingly, and in contrast to UV irradiation or mitomycin C treatment, pressure was not able to induce Stx prophage or an SOS response in several natural Stx-producing STEC isolates. PMID- 15746314 TI - Effect of heat, acidification, and chlorination on Salmonella enterica serovar typhimurium cells in a biofilm formed at the air-liquid interface. AB - Bacterial biofilms have great significance for public health, since biofilm associated microorganisms exhibit dramatically decreased susceptibility to antimicrobial agents and treatments. To date most attention has focused on biofilms that arise from the colonization of solid-liquid or solid-air interfaces. It is of interest that colonization of the interface between air and liquid, which can be selectively advantageous for aerobic or facultative aerobic bacteria, has been rarely studied, although it may present a major problem in industrial aquatic systems. In this work we investigated the role of a biofilm at the interface between air and liquid (pellicle) in the susceptibility of Salmonella enterica serovar Typhimurium to stress conditions. For a control we used a mutant that had lost its ability to synthesize cellulose and thin aggregative fimbriae and thus did not produce the pellicle. Resistance of bacteria from the pellicle to heat, acidification, and chlorination was compared to resistance of planktonic cells from the logarithmic and stationary phases of growth. Pellicle cells were significantly more resistant to chlorination, and thus the surrounding matrix conferred protection against the reactive sodium hypochlorite. However, the stress management of pellicle cells in response to heat and low pH was not enhanced compared to that of stationary-phase cells. A long-period of incubation resulted in endogenous hydrolysis of the pellicle matrix. This phenomenon provides a potential new approach to combat microbial cells in biofilms. PMID- 15746315 TI - Biosynthesis of auxin by the gram-positive phytopathogen Rhodococcus fascians is controlled by compounds specific to infected plant tissues. AB - The role and metabolism of indole-3-acetic acid in gram-negative bacteria is well documented, but little is known about indole-3-acetic acid biosynthesis and regulation in gram-positive bacteria. The phytopathogen Rhodococcus fascians, a gram-positive organism, incites diverse developmental alterations, such as leafy galls, on a wide range of plants. Phenotypic analysis of a leafy gall suggests that auxin may play an important role in the development of the symptoms. We show here for the first time that R. fascians produces and secretes the auxin indole-3 acetic acid. Interestingly, whereas noninfected-tobacco extracts have no effect, indole-3-acetic acid synthesis is highly induced in the presence of infected tobacco extracts when tryptophan is not limiting. Indole-3-acetic acid production by a plasmid-free strain shows that the biosynthetic genes are located on the bacterial chromosome, although plasmid-encoded genes contribute to the kinetics and regulation of indole-3-acetic acid biosynthesis. The indole-3-acetic acid intermediates present in bacterial cells and secreted into the growth media show that the main biosynthetic route used by R. fascians is the indole-3-pyruvic acid pathway with a possible rate-limiting role for indole-3-ethanol. The relationship between indole-3-acetic acid production and the symptoms induced by R. fascians is discussed. PMID- 15746316 TI - Toxin-producing ability among Bacillus spp. outside the Bacillus cereus group. AB - A total of 333 Bacillus spp. isolated from foods, water, and food plants were examined for the production of possible enterotoxins and emetic toxins using a cytotoxicity assay on Vero cells, the boar spermatozoa motility assay, and a liquid chromatography-mass spectrometry method. Eight strains produced detectable toxins; six strains were cytotoxic, three strains produced putative emetic toxins (different in size from cereulide), and one strain produced both cytotoxin(s) and putative emetic toxin(s). The toxin-producing strains could be assigned to four different species, B. subtilis, B. mojavensis, B. pumilus, or B. fusiformis, by using a polyphasic approach including biochemical, chemotaxonomic, and DNA-based analyses. Four of the strains produced cytotoxins that were concentrated by ammonium sulfate followed by dialysis, and two strains produced cytotoxins that were not concentrated by such a treatment. Two cultures maintained full cytotoxic activity, two cultures reduced their activity, and two cultures lost their activity after boiling. The two most cytotoxic strains (both B. mojavensis) were tested for toxin production at different temperatures. One of these strains produced cytotoxin at growth temperatures ranging from 25 to 42 degrees C, and no reduction in activity was observed even after 24 h of growth at 42 degrees C. The strains that produced putative emetic toxins were tested for the influence of time and temperature on the toxin production. It was shown that they produced putative emetic toxin faster or just as fast at 30 as at 22 degrees C. None of the cytotoxic strains produced B. cereus-like enterotoxins as tested by PCR or by immunological methods. PMID- 15746317 TI - Acquisition of resistance to extended-spectrum cephalosporins by Salmonella enterica subsp. enterica serovar Newport and Escherichia coli in the turkey poult intestinal tract. AB - Salmonella enterica subsp. enterica serovar Newport resistant to the extended spectrum cephalosporins (ESCs) and other antimicrobials causes septicemic salmonellosis in humans and animals and is increasingly isolated from humans, animals, foods, and environmental sources. Mechanisms whereby serovar Newport bacteria become resistant to ESCs and other classes of antimicrobials while inhabiting the intestinal tract are not well understood. The present study shows that 25.3% of serovar Newport strains isolated from the turkey poult intestinal tract after the animals were dosed with Escherichia coli harboring a large conjugative plasmid encoding the CMY-2 beta-lactamase and other drug resistance determinants acquired the plasmid and its associated drug resistance genes. The conjugative plasmid containing the cmy-2 gene was transferred not only from the donor E. coli to Salmonella serovar Newport but also to another E. coli serotype present in the intestinal tract. Laboratory studies showed that the plasmid could be readily transferred between serovar Newport and E. coli intestinal isolates. Administration of a single dose of ceftiofur, used to prevent septicemic colibacillosis, to 1-day-old turkeys did not result in the isolation of ceftiofur resistant E. coli or Salmonella serovar Newport. There was a remarkable association between serotype, drug resistance, and plasmid profile among the E. coli strains isolated from the poults. This study shows that Salmonella serovar Newport can become resistant to ESCs and other antibiotics by acquiring a conjugative drug resistance plasmid from E. coli in the intestines. PMID- 15746318 TI - Species abundance and diversity of Burkholderia cepacia complex in the environment. AB - Despite considerable interest in studying Burkholderia cepacia complex in the environment, we still do not have efficient methods to detect, isolate, and screen large numbers of B. cepacia isolates. To better describe the ecology and diversity of B. cepacia complex, a colony hybridization assay was developed to detect specifically all species of the complex based on polymorphism of the variable V3 region of the 16S rRNA sequence. The sensitivity of the assay was dramatically enhanced by using a probe consisting of three repeats of a B. cepacia complex-specific probe, each separated by a phosphoramidite spacer. In addition, a duplex PCR targeting B. cepacia complex-specific recA and 16S rRNA sequences was developed to enable a fast and reliable diagnostic assay for members of the complex. When applied to maize rhizosphere samples, colony hybridization results were in good agreement with those of most-probable-number duplex PCR, both indicating a >100-fold fluctuation of abundance between individual plants. Using restriction analysis of recA for a total of 285 confirmed isolates of the B. cepacia complex, up to seven B. cepacia complex species were identified; however, their diversity and abundance were not evenly distributed among individual plants, and several allelic variants were commonly found from the same rhizosphere sample. These results indicate that not only complex communities of B. cepacia complex species and closely related strains of the same species may coexist at high population levels but also species composition and abundance may dramatically vary between individual plants. PMID- 15746319 TI - Stable-isotope probing of bacteria capable of degrading salicylate, naphthalene, or phenanthrene in a bioreactor treating contaminated soil. AB - [13C6]salicylate, [U-13C]naphthalene, and [U-13C]phenanthrene were synthesized and separately added to slurry from a bench-scale, aerobic bioreactor used to treat soil contaminated with polycyclic aromatic hydrocarbons. Incubations were performed for either 2 days (salicylate, naphthalene) or 7 days (naphthalene, phenanthrene). Total DNA was extracted from the incubations, the "heavy" and "light" DNA were separated, and the bacterial populations associated with the heavy fractions were examined by denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone libraries. Unlabeled DNA from Escherichia coli K-12 was added to each sample as an internal indicator of separation efficiency. While E. coli was not detected in most analyses of heavy DNA, a low number of E. coli sequences was recovered in the clone libraries associated with the heavy DNA fraction of [13C]phenanthrene incubations. The number of E. coli clones recovered proved useful in determining the relative amount of light DNA contamination of the heavy fraction in that sample. Salicylate- and naphthalene-degrading communities displayed similar DGGE profiles and their clone libraries were composed primarily of sequences belonging to the Pseudomonas and Ralstonia genera. In contrast, heavy DNA from the phenanthrene incubations displayed a markedly different DGGE profile and was composed primarily of sequences related to the Acidovorax genus. There was little difference in the DGGE profiles and types of sequences recovered from 2- and 7-day incubations with naphthalene, so secondary utilization of the 13C during the incubation did not appear to be an issue in this experiment. PMID- 15746320 TI - Mycobacterium avium subspecies paratuberculosis cultured from locally and commercially pasteurized cow's milk in the Czech Republic. AB - Between November 2002 and April 2003, 244 bottles and cartons of commercially pasteurized cow's milk were obtained at random from retail outlets throughout the Czech Republic. During the same period, samples of raw milk and of milk that was subsequently subjected to a minimum of 71.7 degrees C for 15 s in a local pasteurization unit were also obtained from two dairy herds, designated herds A and B, with low and high levels, respectively, of subclinical Mycobacterium avium subsp. paratuberculosis infection, and from one herd, herd C, without infection. Infection in individual cows in each herd was tested by fecal culturing. Milk samples were brought to the Veterinary Research Institute in Brno, Czech Republic, processed, inoculated onto Herrold's egg yolk slants, and incubated for 32 weeks. Colonies were characterized by morphology, Ziehl-Neelsen staining, mycobactin J dependency, and IS900 PCR results. M. avium subsp. paratuberculosis was cultured from 4 of 244 units (1.6%) of commercially pasteurized retail milk. M. avium subsp. paratuberculosis was also cultured from 2 of 100 (2%) cartons of locally pasteurized milk derived from infected herds A and B and from 0 of 100 cartons of milk from uninfected herd C. Raw milk from 1 of 10 (10%) fecal culture positive cows in herd A and from 13 of 66 (19.7%) fecal culture-positive cows in herd B was culture positive for M. avium subsp. paratuberculosis. These findings confirm that M. avium subsp. paratuberculosis is present in raw milk from subclinically infected dairy cows. The culture of M. avium subsp. paratuberculosis in the Czech Republic from retail milk that had been pasteurized locally or commercially to the required national and European Union standards is in agreement with similar research on milk destined for consumers in the United Kingdom and the United States and shows that humans are being exposed to this chronic enteric pathogen by this route. PMID- 15746321 TI - Heterologous production, assembly, and secretion of a minicellulosome by Clostridium acetobutylicum ATCC 824. AB - The gene man5K encoding the mannanase Man5K from Clostridium cellulolyticum was cloned alone or as an operon with the gene cipC1 encoding a truncated scaffoldin (miniCipC1) of the same origin in the solventogenic Clostridium acetobutylicum. The expression of the heterologous gene(s) was under the control of a weakened thiolase promoter Pthl. The recombinant strains of the solventogenic bacterium were both found to secrete active Man5K in the range of milligrams per liter. In the case of the strain expressing only man5K, a large fraction of the recombinant enzyme was truncated and lost the N-terminal dockerin domain, but it remained active towards galactomannan. When man5K was coexpressed with cipC1 in C. acetobutylicum, the recombinant strain secreted almost exclusively full-length mannanase, which bound to the scaffoldin miniCipC1, thus showing that complexation to the scaffoldin stabilized the enzyme. The secreted heterologous complex was found to be functional: it binds to crystalline cellulose via the carbohydrate binding module of the miniscaffoldin, and the complexed mannanase is active towards galactomannan. Taken together, these data show that C. acetobutylicum is a suitable host for the production, assembly, and secretion of heterologous minicellulosomes. PMID- 15746323 TI - Analyses of livestock production, waste storage, and pathogen levels and prevalences in farm manures. AB - Survey results describing the levels and prevalences of zoonotic agents in 1,549 livestock waste samples were analyzed for significance with livestock husbandry and farm waste management practices. Statistical analyses of survey data showed that livestock groups containing calves of <3 months of age, piglets, or lambs had higher prevalences and levels of Campylobacter spp. and Escherichia coli O157 in their wastes. Younger calves that were still receiving milk, however, had significantly lower levels and prevalence of E. coli O157. Furthermore, when wastes contained any form of bedding, they had lowered prevalences and levels of both pathogenic Listeria spp. and Campylobacter spp. Livestock wastes generated by stock consuming a diet composed principally of grass were less likely to harbor E. coli O157 or Salmonella spp. Stocking density did not appear to influence either the levels or prevalences of bacterial pathogens. Significant seasonal differences in prevalences were detected in cattle wastes; Listeria spp. were more likely to be isolated in March to June, and E. coli O157 was more likely to be found in May and June. Factors such as livestock diet and age also had significant influence on the levels and prevalences of some zoonotic agents in livestock wastes. A number of the correlations identified could be used as the basis of a best-practice disposal document for farmers, thereby lowering the microbiological risks associated with applying manures of contaminated livestock to land. PMID- 15746322 TI - Functional analysis of three plasmids from Lactobacillus plantarum. AB - Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism. The host range of the pWCFS101 replicon includes Lactobacillus species and Lactococcus lactis, while that of the pWCFS102 replicon also includes Carnobacterium maltaromaticum and Bacillus subtilis. The larger plasmid is predicted to replicate via the theta type mechanism. The host range of its replicon seems restricted to L. plantarum. Cloning vectors were constructed based on the replicons of all three plasmids. Plasmid pWCFS103 was demonstrated to be a conjugative plasmid, as it could be transferred to L. plantarum NC8. It confers arsenate and arsenite resistance, which can be used as selective markers. PMID- 15746324 TI - Characterization of the cro-ori region of the Streptococcus thermophilus virulent bacteriophage DT1. AB - The virulent cos-type Streptococcus thermophilus phage DT1 was previously isolated from a mozzarella whey sample, and its complete genomic sequence is available. The putative ori of phage DT1 is characterized by three inverted and two direct repeats located in a noncoding region between orf36 and orf37. As the replication ability of the putative ori and flanking genes could not be established, its ability to confer phage resistance was tested. When ori is cloned on a high-copy-number plasmid, it provides protection to S. thermophilus strains against phage infection during milk fermentation. This protection is phage specific and strain dependent. Then, a detailed transcriptional map was established for the region located between the cro-like gene (orf29) and the ori. The results of the Northern blots indicated that the transcription of this region started 5 min after the onset of phage infection. Comparative analysis of the expression of the cro-ori region in the three S. thermophilus cos-type phages DT1, Sfi19 (virulent), and Sfi21 (temperate) reveals significant differences in the number and size of transcripts. The promoter upstream of orf29 was further investigated by primer extension analysis, and its activity was confirmed by a chloramphenicol acetyltransferase assay, which showed that the phage promoter is more efficient than the constitutive bacterial promoter of the S. thermophilus operon encoding the general proteins of the phosphoenolpyruvate:sugar phosphotransferase system. However, the phage promoter is less efficient than the pts promoter in Lactococcus lactis and in Escherichia coli. PMID- 15746325 TI - Degradation of wheat straw by Fibrobacter succinogenes S85: a liquid- and solid state nuclear magnetic resonance study. AB - Wheat straw degradation by Fibrobacter succinogenes was monitored by nuclear magnetic resonance (NMR) spectroscopy and chemolytic methods to investigate the activity of an entire fibrolytic system on an intact complex substrate. In situ solid-state NMR with 13C cross-polarization magic angle spinning was used to monitor the modification of the composition and structure of lignocellulosic fibers (of 13C-enriched wheat straw) during the growth of bacteria on this substrate. There was no preferential degradation either of amorphous regions of cellulose versus crystalline regions or of cellulose versus hemicelluloses in wheat straw. This suggests either a simultaneous degradation of the amorphous and crystalline parts of cellulose and of cellulose and hemicelluloses by the enzymes or degradation at the surface at a molecular scale that cannot be detected by NMR. Liquid-state two-dimensional NMR experiments and chemolytic methods were used to analyze in detail the various sugars released into the culture medium. An integration of NMR signals enabled the quantification of oligosaccharides produced from wheat straw at various times of culture and showed the sequential activities of some of the fibrolytic enzymes of F. succinogenes S85 on wheat straw. In particular, acetylxylan esterase appeared to be more active than arabinofuranosidase, which was more active than alpha-glucuronidase. Finally, cellodextrins did not accumulate to a great extent in the culture medium. PMID- 15746326 TI - Degradation of 1,4-dioxane and cyclic ethers by an isolated fungus. AB - By using 1,4-dioxane as the sole source of carbon, a 1,4-dioxane-degrading microorganism was isolated from soil. The fungus, termed strain A, was able to utilize 1,4-dioxane and many kinds of cyclic ethers as the sole source of carbon and was identified as Cordyceps sinensis from its 18S rRNA gene sequence. Ethylene glycol was identified as a degradation product of 1,4-dioxane by the use of deuterated 1,4-dioxane-d8 and gas chromatography-mass spectrometry analysis. A degradation pathway involving ethylene glycol, glycolic acid, and oxalic acid was proposed, followed by incorporation of the glycolic acid and/or oxalic acid via glyoxylic acid into the tricarboxylic acid cycle. PMID- 15746327 TI - Enumeration and diversity of campylobacters and bacteriophages isolated during the rearing cycles of free-range and organic chickens. AB - Campylobacters and Campylobacter-specific bacteriophages were isolated and enumerated during the rearing cycle of free-range (56 days) and organic chickens (73 days) at 3-day intervals from hatching until slaughter. In both flocks Campylobacter jejuni was the initial colonizer but Campylobacter coli was detected more frequently from 5 weeks of age. The diversity of the Campylobacter isolates was examined by pulsed-field gel electrophoresis of SmaI-digested genomic DNA and antimicrobial resistance typing. Bacteriophages were isolated from 51% (19 of 37 birds) of Campylobacter-positive organic birds (log10 2.5 to log10 5.7 PFU/g of cecal contents). The bacteriophages were all typical group III Campylobacter bacteriophages in terms of genomic size but could be characterized in terms of their host range and placed into five different groups. In contrast to the organic birds, anti-Campylobacter activity (bacteriocin-like) was observed in 26% (10 of 38 birds) of Campylobacter-positive free-range birds, and only one bacteriophage was isolated. Appearance of either bacteriophages or anti Campylobacter activity was associated with changes in the levels of colonization and the predominant genotypes and species isolated. The frequency and potential influence of naturally occurring bacteriophages and/or inhibitory substances on the diversity and fluctuations of populations of campylobacters have not previously been reported in either free-range or organic chickens. PMID- 15746328 TI - Poly(A) polymerase modification and reverse transcriptase PCR amplification of environmental RNA. AB - We describe a combination of two established techniques for a novel application for constructing full-length cDNA clone libraries from environmental RNA. The cDNA was cloned without the use of prescribed primers that target specific genes, and the procedure did not involve random priming. Purified RNA was first modified by addition of a poly(A) tail and then was amplified by using a commercially available reverse transcriptase PCR (RT-PCR) cDNA synthesis kit. To demonstrate the feasibility of this approach, a cDNA clone library was constructed from size fractionated RNA (targeting 16S rRNA) purified from a geothermally heated soil in Yellowstone National Park in Wyoming. The resulting cDNA library contained clones representing Bacteria and Eukarya taxa and several mRNAs. There was no exact clone match between this library and a separate cDNA library generated from an RT PCR performed with unmodified rRNA and Bacteria-specific forward and universal reverse primers that were designed from cultivated organisms; however, both libraries contained representatives of the Firmicutes and the alpha Proteobacteria. Unexpectedly, there were no Archaea clones in the library generated from poly(A)-modified RNA. Additional RT-PCRs performed with universal and Archaea-biased primers and unmodified RNA demonstrated the presence of novel Archaea in the soil. Experiments with pure cultures of Sulfolobus solfataricus and Halobacterium halobium revealed that some Archaea rRNA may not be a suitable substrate for the poly(A) tail modification step. The protocol described here demonstrates the feasibility of directly accessing prokaryote RNA (rRNA and/or mRNA) in environmental samples, but the results also illustrate potentially important problems. PMID- 15746329 TI - Influence of starvation on potential ammonia-oxidizing activity and amoA mRNA levels of Nitrosospira briensis. AB - The effect of short-term ammonia starvation on Nitrosospira briensis was investigated. The ammonia-oxidizing activity was determined in a concentrated cell suspension with a NOx biosensor. The apparent half-saturation constant [Km(app)] value of the NH3 oxidation of N. briensis was 3 microM NH3 for cultures grown both in continuous and batch cultures as determined by a NOx biosensor. Cells grown on the wall of the vessel had a lower Km(app) value of 1.8 microM NH3. Nonstarving cultures of N. briensis showed potential ammonia-oxidizing activities of between 200 to 250 microM N h(-1), and this activity decreased only slowly during starvation up to 10 days. Within 10 min after the addition of fresh NH4+, 100% activity was regained. Parallel with activity measurements, amoA mRNA and 16S rRNA were investigated. No changes were observed in the 16S rRNA, but a relative decrease of amoA mRNA was observed during the starvation period. During resuscitation, an increase in amoA mRNA expression was detected simultaneously. The patterns of the soluble protein fraction of a 2-week-starved culture of N. briensis showed only small differences in comparison to a nonstarved control. From these results we conclude that N. briensis cells remain in a state allowing fast recovery of ammonia-oxidizing activity after addition of NH4+ to a starved culture. Maintaining cells in this kind of active state could be the survival strategy of ammonia-oxidizing bacteria in nature under fluctuating NH4+ availability. PMID- 15746330 TI - Novel activator of mannose-specific phosphotransferase system permease expression in Listeria innocua, identified by screening for pediocin AcH resistance. AB - To identify genes that are important for class IIa bacteriocin interaction and resistance in Listeria species, transposon Tn917 knockout libraries were constructed for Listeria innocua strain Lin11 and screened for mutants that are resistant to pediocin AcH. A highly resistant mutant (G7) (MIC > 20 microg/ml; 1,000-fold less susceptible than the wild type), in which the transposon integrated into the putative promoter of the lin0142 gene, was isolated. lin0142 is located immediately upstream of the mpt operon (mptA/mptC/mptD) that encodes the mannose-specific phosphoenolpyruvate-dependent phosphotransferase system permease EIItMan, which serves as a docking protein for class IIa bacteriocins. The transcription of the mpt operon is known to be positively controlled by sigma54 factor and ManR (a sigma54-associated activator). Transcripts for lin0142 and mpt were undetectable in the G7 mutant, based on quantitative real-time reverse transcriptase PCR analysis. When the wild-type lin0142 gene was expressed at a 7.9-fold-elevated level in the mutant via a multicopy-number plasmid, the level of mpt mRNA became 70% higher than that in the wild-type strain. In addition, the complementation strain reverted back to the pediocin AcH susceptible phenotype. The levels of manR and rpoN (sigma54) mRNAs were not directly influenced by the level of lin0142 transcription. lin0142 is the only one of the three mpt regulatory genes whose transcription is induced, albeit slightly (1.2-fold), by glucose. The combined results show that the lin0142 gene encodes a novel activator of the mpt operon. The Lin0142 protein contains a winged-helix DNA-binding motif and is distantly related to the Crp-Fnr family of transcription regulators. PMID- 15746331 TI - Rapid acyl-homoserine lactone quorum signal biodegradation in diverse soils. AB - Signal degradation impacts all communications. Although acyl-homoserine lactone (acyl-HSL) quorum-sensing signals are known to be degraded by defined laboratory cultures, little is known about their stability in nature. Here, we show that acyl-HSLs are biodegraded in soils sampled from diverse U.S. sites and by termite hindgut contents. When amended to samples at physiologically relevant concentrations, 14C-labeled acyl-HSLs were mineralized to 14CO2 rapidly and, at most sites examined, without lag. A lag-free turf soil activity was characterized in further detail. Heating or irradiation of the soil prior to the addition of radiolabel abolished mineralization, whereas protein synthesis inhibitors did not. Mineralization exhibited an apparent Km of 1.5 microM acyl-HSL, ca. 1,000 fold lower than that reported for a purified acyl-HSL lactonase. Under optimal conditions, acyl-HSL degradation proceeded at a rate of 13.4 nmol x h(-1) x g of fresh weight soil(-1). Bioassays established that the final extent of signal inactivation was greater than for its full conversion to CO2 but that the two processes were well coupled kinetically. A most probable number of 4.6 x 10(5) cells . g of turf soil(-1) degraded physiologically relevant amounts of hexanoyl [1-14C]HSL to 14CO2. It would take chemical lactonolysis months to match the level of signal decay achieved in days by the observed biological activity. Rapid decay might serve either to quiet signal cross talk that might otherwise occur between spatially separated microbial aggregates or as a full system reset. Depending on the context, biological signal decay might either promote or complicate cellular communications and the accuracy of population density-based controls on gene expression in species-rich ecosystems. PMID- 15746332 TI - Spatially resolved characterization of biogenic manganese oxide production within a bacterial biofilm. AB - Pseudomonas putida strain MnB1, a biofilm-forming bacterial culture, was used as a model for the study of bacterial Mn oxidation in freshwater and soil environments. The oxidation of aqueous Mn+2 [Mn+2(aq)] by P. putida was characterized by spatially and temporally resolving the oxidation state of Mn in the presence of a bacterial biofilm, using scanning transmission X-ray microscopy (STXM) combined with near-edge X-ray absorption fine structure (NEXAFS) spectroscopy at the Mn L2,3 absorption edges. Subsamples were collected from growth flasks containing 0.1 and 1 mM total Mn at 16, 24, 36, and 48 h after inoculation. Immediately after collection, the unprocessed hydrated subsamples were imaged at a 40-nm resolution. Manganese NEXAFS spectra were extracted from X ray energy sequences of STXM images (stacks) and fit with linear combinations of well-characterized reference spectra to obtain quantitative relative abundances of Mn(II), Mn(III), and Mn(IV). Careful consideration was given to uncertainty in the normalization of the reference spectra, choice of reference compounds, and chemical changes due to radiation damage. The STXM results confirm that Mn+2(aq) was removed from solution by P. putida and was concentrated as Mn(III) and Mn(IV) immediately adjacent to the bacterial cells. The Mn precipitates were completely enveloped by bacterial biofilm material. The distribution of Mn oxidation states was spatially heterogeneous within and between the clusters of bacterial cells. Scanning transmission X-ray microscopy is a promising tool for advancing the study of hydrated interfaces between minerals and bacteria, particularly in cases where the structure of bacterial biofilms needs to be maintained. PMID- 15746333 TI - Diversity of proteolytic Clostridium botulinum strains, determined by a pulsed field gel electrophoresis approach. AB - Pulsed-field gel electrophoresis (PFGE) was applied to the study of the similarity of 55 strains of proteolytic Clostridium botulinum (C. botulinum group I) types A, AB, B, and F. Rare-cutting restriction enzymes ApaI, AscI, MluI, NruI, PmeI, RsrII, SacII, SmaI, and XhoI were tested for their suitability for the cleavage of DNA of five proteolytic C. botulinum strains. Of these enzymes, SacII, followed by SmaI and XhoI, produced the most convenient number of fragments for genetic typing and were selected for analysis of the 55 strains. The proteolytic C. botulinum species was found to be heterogeneous. In the majority of cases, PFGE enabled discrimination between individual strains of proteolytic C. botulinum types A and B. The different toxin types were discriminated at an 86% similarity level with both SacII and SmaI and at an 83% similarity level with XhoI. Despite the high heterogeneity, three clusters at a 95% similarity level consisting of more than three strains of different origin were noted. The strains of types A and B showed higher diversity than the type F organisms which formed a single cluster. According to this survey, PFGE is to be considered a useful tool for molecular epidemiological analysis of proteolytic C. botulinum types A and B. However, epidemiological conclusions based on PFGE data only should be made with discretion, since highly similar PFGE patterns were noticed, especially within the type B strains. PMID- 15746334 TI - Nodulation of Lupinus albus by strains of Ochrobactrum lupini sp. nov. AB - The nodulation of legumes has for more than a century been considered an exclusive capacity of a group of microorganisms commonly known as rhizobia and belonging to the alpha-Proteobacteria. However, in the last 3 years four nonrhizobial species, belonging to alpha and beta subclasses of the Proteobacteria, have been described as legume-nodulating bacteria. In the present study, two fast-growing strains, LUP21 and LUP23, were isolated from nodules of Lupinus honoratus. The phylogenetic analysis based on the 16S and 23S rRNA gene sequences showed that the isolates belong to the genus Ochrobactrum. The strains were able to reinfect Lupinus plants. A plasmid profile analysis showed the presence of three plasmids. The nodD and nifH genes were located on these plasmids, and their sequences were obtained. These sequences showed a close resemblance to the nodD and nifH genes of rhizobial species, suggesting that the nodD and nifH genes carried by strain LUP21T were acquired by horizontal gene transfer. A polyphasic study including phenotypic, chemotaxonomic, and molecular features of the strains isolated in this study showed that they belong to a new species of the genus Ochrobactrum for which we propose the name Ochrobactrum lupini sp. nov. Strain LUP21T (LMG 20667T) is the type strain. PMID- 15746335 TI - Rhizobial 16S rRNA and dnaK genes: mosaicism and the uncertain phylogenetic placement of Rhizobium galegae. AB - The phylogenetic relatedness among 12 agriculturally important species in the order Rhizobiales was estimated by comparative 16S rRNA and dnaK sequence analyses. Two groups of related species were identified by neighbor-joining and maximum-parsimony analysis. One group consisted of Mesorhizobium loti and Mesorhizobium ciceri, and the other group consisted of Agrobacterium rhizogenes, Rhizobium tropici, Rhizobium etli, and Rhizobium leguminosarum. Although bootstrap support for the placement of the remaining six species varied, A. tumefaciens, Agrobacterium rubi, and Agrobacterium vitis were consistently associated in the same subcluster. The three other species included Rhizobium galegae, Sinorhizobium meliloti, and Brucella ovis. Among these, the placement of R. galegae was the least consistent, in that it was placed flanking the A. rhizogenes-Rhizobium cluster in the dnaK nucleotide sequence trees, while it was placed with the other three Agrobacterium species in the 16S rRNA and the DnaK amino acid trees. In an effort to explain the inconsistent placement of R. galegae, we examined polymorphic site distribution patterns among the various species. Localized runs of nucleotide sequence similarity were evident between R. galegae and certain other species, suggesting that the R. galegae genes are chimeric. These results provide a tenable explanation for the weak statistical support often associated with the phylogenetic placement of R. galegae, and they also illustrate a potential pitfall in the use of partial sequences for species identification. PMID- 15746337 TI - Fingerprinting of Bacillus thuringiensis type strains and isolates by using Bacillus cereus group-specific repetitive extragenic palindromic sequence-based PCR analysis. AB - A total of 119 Bacillus thuringiensis strains (83 type strains and 26 native isolates), as well as five B. cereus group species, were analyzed by repetitive extragenic palindromic sequence-based PCR analysis (Rep-PCR) fingerprinting. Primers Bc-REP-1 and Bc-REP-2 were specifically designed according to an extragenic 26-bp repeated sequence found in the six B. cereus group genomes reported. A total of 47 polymorphic bands were detected, and the patterns varied from 5 to 13 bands in number and from 0.2 to 3.8 kb in size. Virtually each type strain showed a distinctive B. cereus (Bc)-Rep-PCR pattern, except for B. thuringiensis serovars dakota (H serotype 15 [H15]) and sotto (H4a,4b), as well as serovars amagiensis (H29) and seoulensis (H35), which shared the same patterns. As expected, serovar entomocidus (H6) and its biovar subtoxicus showed an identical pattern; similarly, serovars sumiyoshiensis (H3a,3d) and fukuokaensis (H3a,3d,3e), which share two antigenic determinants, also showed identical Bc-Rep-PCR patterns. Interestingly, serovars israelensis (H14) and malaysiensis (H36), which share several phenotypic attributes, also showed identical Bc-Rep-PCR patterns. Native, coleopteran-active strains, including the self-agglutinated LBIT-74 strain, showed Bc-Rep-PCR patterns identical or very similar to that of the tenebrionis strain. Likewise, native mosquitocidal strains (including some self-agglutinated strains) also showed patterns identical or very similar to that of the serovar israelensis IPS-82 strain. Additionally, native beta-exotoxin-producing strains from serovar thuringiensis showed patterns identical to that of the B. thuringiensis type strain. The B. cereus group specific Bc-Rep-PCR fingerprinting technique was shown to be highly discriminative, fast, easy, and able to identify B. thuringiensis serotypes, including nonflagellar and self-agglutinated strains. PMID- 15746336 TI - Identification of a new Borrelia species among small mammals in areas of northern Spain where Lyme disease is endemic. AB - The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several areas where Lyme disease is endemic in northern Spain. A low rate of infestation by Ixodes ricinus nymphs was found in the small mammal populations studied that correlated with the near-absence of B. burgdorferi sensu lato in 184 animals tested and with the lack of transmission of B. burgdorferi sensu lato to I. ricinus larvae that fed on them. In contrast, questing ticks collected at the same time and in the same areas were found to carry a highly variable B. burgdorferi sensu lato repertoire (B. burgdorferi sensu stricto, Borrelia garinii, Borrelia valaisiana, and Borrelia afzelii). Interestingly, the only isolate obtained from small mammals (R57, isolated from a bank vole) grouped by phylogenetic analyses with other Borrelia species but in a separate clade from the Lyme disease and relapsing fever organisms, suggesting that it is a new species. This new agent was widely distributed among small mammals, with infection rates of 8.5 to 12% by PCR. Moreover, a high seroprevalence to B. burgdorferi sensu lato was found in the animal sera, suggesting cross-reactivity between B. burgdorferi sensu lato and R57. Although small mammals do not seem to play an important role as reservoirs for B. burgdorferi sensu lato in the study area, they seem to be implicated in the maintenance of spirochetes similar to R57. PMID- 15746338 TI - Differential activities of four Lactobacillus casei promoters during bacterial transit through the gastrointestinal tracts of human-microbiota-associated mice. AB - In a previous study using fusion of the deregulated lactose promoter lacTp* and reporter genes, we suggested that Lactobacillus casei could initiate de novo protein synthesis during intestinal transit. In order to confirm this finding and extend it to other promoters, we adopted a reverse transcriptase quantitative PCR (RT-QPCR) approach combined with a transcriptional fusion system consisting of luciferase genes under the control of four promoters (ccpA, dlt, ldh, and lacT*) from L. casei DN-114 001. Promoter expression was monitored during cell growth, and variable luciferase activities were detected. In 3-day cultures, all the genetically modified strains survived but without exhibiting luciferase activity. Luciferase mRNA levels determined by RT-QPCR analysis (RNA/CFU) were not significant. The cultures were administered to human-microbiota-associated mice, and the feces were collected 6 h later. L. casei promoters lacTp* and ldhp initiated mRNA synthesis during gastrointestinal transit. The promoters, ccpAp and dltp, exhibited no luciferase activity, nor was de novo-synthesized luciferase mRNA detected in the feces. L. casei seems to adapt its physiology to the gastrointestinal tract environment by modulating promoter activities. The approach (fecal transcriptional analysis) described herein may, moreover, be of value in studying gene expression of transiting bacteria in human fecal specimens. PMID- 15746339 TI - The doubly phosphorylated form of HPr, HPr(Ser~P)(His-P), is abundant in exponentially growing cells of Streptococcus thermophilus and phosphorylates the lactose transporter LacS as efficiently as HPr(His~P). AB - In Streptococcus thermophilus, lactose is taken up by LacS, a transporter that comprises a membrane translocator domain and a hydrophilic regulatory domain homologous to the IIA proteins and protein domains of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). The IIA domain of LacS (IIALacS) possesses a histidine residue that can be phosphorylated by HPr(His~P), a protein component of the PTS. However, determination of the cellular levels of the different forms of HPr, namely, HPr, HPr(His~P), HPr(Ser-P), and HPr(Ser P)(His~P), in exponentially lactose-growing cells revealed that the doubly phosphorylated form of HPr represented 75% and 25% of the total HPr in S. thermophilus ATCC 19258 and S. thermophilus SMQ-301, respectively. Experiments conducted with [32P]PEP and purified recombinant S. thermophilus ATCC 19258 proteins (EI, HPr, and IIALacS) showed that IIALacS was reversibly phosphorylated by HPr(Ser-P)(His~P) at a rate similar to that measured with HPr(His~P). Sequence analysis of the IIALacS protein domains from several S. thermophilus strains indicated that they can be divided into two groups on the basis of their amino acid sequences. The amino acid sequence of IIALacS from group I, to which strain 19258 belongs, differed from that of group II at 11 to 12 positions. To ascertain whether IIALacS from group II could also be phosphorylated by HPr(His~P) and HPr(Ser-P)(His~P), in vitro phosphorylation experiments were conducted with purified proteins from Streptococcus salivarius ATCC 25975, which possesses a IIALacS very similar to group II S. thermophilus IIALacS. The results indicated that S. salivarius IIALacS was phosphorylated by HPr(Ser-P)(His~P) at a higher rate than that observed with HPr(His~P). Our results suggest that the reversible phosphorylation of IIALacS in S. thermophilus is accomplished by HPr(Ser P)(His~P) as well as by HPr(His~P). PMID- 15746340 TI - 16S rRNA gene-based oligonucleotide microarray for environmental monitoring of the betaproteobacterial order "Rhodocyclales". AB - For simultaneous identification of members of the betaproteobacterial order "Rhodocyclales" in environmental samples, a 16S rRNA gene-targeted oligonucleotide microarray (RHC-PhyloChip) consisting of 79 probes was developed. Probe design was based on phylogenetic analysis of available 16S rRNA sequences from all cultured and as yet uncultured members of the "Rhodocyclales." The multiple nested probe set was evaluated for microarray hybridization with 16S rRNA gene PCR amplicons from 29 reference organisms. Subsequently, the RHC PhyloChip was successfully used for cultivation-independent "Rhodocyclales" diversity analysis in activated sludge from an industrial wastewater treatment plant. The implementation of a newly designed "Rhodocyclales"-selective PCR amplification system prior to microarray hybridization greatly enhanced the sensitivity of the RHC-PhyloChip and thus enabled the detection of "Rhodocyclales" populations with relative abundances of less than 1% of all bacteria (as determined by fluorescence in situ hybridization) in the activated sludge. The presence of as yet uncultured Zoogloea-, Ferribacterium/Dechloromonas , and Sterolibacterium-related bacteria in the industrial activated sludge, as indicated by the RHC-PhyloChip analysis, was confirmed by retrieval of their 16S rRNA gene sequences and subsequent phylogenetic analysis, demonstrating the suitability of the RHC-PhyloChip as a novel monitoring tool for environmental microbiology. PMID- 15746341 TI - Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. AB - Stenotrophomonas maltophilia is one of the most prevalent opportunistic bacteria causing nosocomial infections. It has become problematic because most of the isolates are resistant to multiple antibiotics, and therefore, development of phage therapy has attracted strong attention. In this study, eight S. maltophilia phages were isolated from clinical samples including patient specimens, catheter related devices, and wastewater. These phages can be divided into four distinct groups based on host range and digestibility of the phage DNAs with different restriction endonucleases. One of them, designated phiSMA5, was further characterized. Electron microscopy showed it resembled Myoviridae, with an isometric head (90 nm in diameter), a tail (90 nm long), a baseplate (25 nm wide), and short tail fibers. The phiSMA5 double-stranded DNA, refractory to digestion by most restriction enzymes, was tested and estimated to be 250 kb by pulsed-field gel electrophoresis. This genome size is second to that of the largest phage, phiKZ of Pseudomonas aeruginosa. In sodium dodecyl sulfate polyacrylamide gel electrophoresis, 25 virion proteins were visualized. N terminal sequencing of four of them suggested that each of them might have had its N terminus cleaved off. Among the 87 S. maltophilia strains collected in this study, only 61 were susceptible to phiSMA5, indicating that more phages are needed toward a phage therapy strategy. Since literature search yielded no information about S. maltophilia phages, phiSMA5 appears to be the first reported. PMID- 15746342 TI - Patterns of antimicrobial resistance observed in Escherichia coli isolates obtained from domestic- and wild-animal fecal samples, human septage, and surface water. AB - A repeated cross-sectional study was conducted to determine the patterns of antimicrobial resistance in 1,286 Escherichia coli strains isolated from human septage, wildlife, domestic animals, farm environments, and surface water in the Red Cedar watershed in Michigan. Isolation and identification of E. coli were done by using enrichment media, selective media, and biochemical tests. Antimicrobial susceptibility testing by the disk diffusion method was conducted for neomycin, gentamicin, streptomycin, chloramphenicol, ofloxacin, trimethoprim sulfamethoxazole, tetracycline, ampicillin, nalidixic acid, nitrofurantoin, cephalothin, and sulfisoxazole. Resistance to at least one antimicrobial agent was demonstrated in isolates from livestock, companion animals, human septage, wildlife, and surface water. In general, E. coli isolates from domestic species showed resistance to the largest number of antimicrobial agents compared to isolates from human septage, wildlife, and surface water. The agents to which resistance was demonstrated most frequently were tetracycline, cephalothin, sulfisoxazole, and streptomycin. There were similarities in the patterns of resistance in fecal samples and farm environment samples by animal, and the levels of cephalothin-resistant isolates were higher in farm environment samples than in fecal samples. Multidrug resistance was seen in a variety of sources, and the highest levels of multidrug-resistant E. coli were observed for swine fecal samples. The fact that water sample isolates were resistant only to cephalothin may suggest that the resistance patterns for farm environment samples may be more representative of the risk of contamination of surface waters with antimicrobial agent-resistant bacteria. PMID- 15746343 TI - Flow-cytometric cell sorting and subsequent molecular analyses for culture independent identification of bacterioplankton involved in dimethylsulfoniopropionate transformations. AB - Marine bacterioplankton transform dimethylsulfoniopropionate (DMSP) into the biogeochemically important and climatically active gas dimethylsulfide. In order to identify specific bacterial taxa mediating DMSP processing in a natural marine ecosystem, we amended water samples from a southeastern U.S. salt marsh with 20 microM DMSP and tracked community shifts with flow cytometry (FCM) coupled to 16S rRNA gene analyses. In two out of four seasons studied, DMSP amendments induced the formation of distinct bacterioplankton populations with elevated nucleic acid (NA) content within 24 h, indicative of cells actively utilizing DMSP. The 16S rRNA genes of the cells with and without elevated NA content were analyzed following cell sorting and PCR amplification with sequencing and terminal restriction fragment length polymorphism approaches. Compared to cells in the control FCM populations, bacteria with elevated NA content in the presence of DMSP were relatively enriched in taxa related to Loktanella, Oceanicola, and Sulfitobacter (Roseobacter lineage, alpha-Proteobacteria); Caulobacter (alpha Proteobacteria); and Brachymonas and Xenophilus (beta-Proteobacteria) in the May 02 sample and to Ketogulonicigenium (Roseobacter lineage, alpha-Proteobacteria) and novel gamma-Proteobacteria in the Sept-02 sample. Our study suggests that diverse bacterioplankton participate in the metabolism of DMSP in coastal marine systems and that their relative importance varies temporally. PMID- 15746344 TI - Histamine-producing pathway encoded on an unstable plasmid in Lactobacillus hilgardii 0006. AB - Histamine production from histidine in fermented food products by lactic acid bacteria results in food spoilage and is harmful to consumers. We have isolated a histamine-producing lactic acid bacterium, Lactobacillus hilgardii strain IOEB 0006, which could retain or lose the ability to produce histamine depending on culture conditions. The hdcA gene, coding for the histidine decarboxylase of L. hilgardii IOEB 0006, was located on an 80-kb plasmid that proved to be unstable. Sequencing of the hdcA locus disclosed a four-gene cluster encoding the histidine decarboxylase, a protein of unknown function, a histidyl-tRNA synthetase, and a protein, which we named HdcP, showing similarities to integral membrane transporters driving substrate/product exchange. The gene coding for HdcP was cloned downstream of a sequence specifying a histidine tag and expressed in Lactococcus lactis. The recombinant HdcP could drive the uptake of histidine into the cell and the exchange of histidine and histamine. The combination of HdcP and the histidine decarboxylase forms a typical bacterial decarboxylation pathway that may generate metabolic energy or be involved in the acid stress response. Analyses of sequences present in databases suggest that the other two proteins have dispensable functions. These results describe for the first time the genes encoding a histamine-producing pathway and provide clues to the parsimonious distribution and the instability of histamine-producing lactic acid bacteria. PMID- 15746345 TI - Posttranslational modification of 6-phosphofructo-1-kinase in Aspergillus niger. AB - Two different enzymes exhibiting 6-phosphofructo-1-kinase (PFK1) activity were isolated from the mycelium of Aspergillus niger: the native enzyme with a molecular mass of 85 kDa, which corresponded to the calculated molecular mass of the deduced amino acid sequence of the A. niger pfkA gene, and a shorter protein of approximately 49 kDa. A fragment of identical size also was obtained in vitro by the proteolytic digestion of the partially purified native PFK1 with proteinase K. When PFK1 activity was measured during the proteolytic degradation of the native protein, it was found to be lost after 1 h of incubation, but it was reestablished after induction of phosphorylation by adding the catalytic subunit of cyclic AMP-dependent protein kinase to the system. By determining kinetic parameters, different ratios of activities measured at ATP concentrations of 0.1 and 1 mM were detected with fragmented PFK1, as with the native enzyme. Fructose-2,6-biphosphate significantly increased the Vmax of the fragmented protein, while it had virtually no effect on the native protein. The native enzyme could be purified only from the early stages of growth on a minimal medium, while the 49-kDa fragment appeared later and was activated at the time of a sudden change in the growth rate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of sequential purifications of PFK1 enzymes by affinity chromatography during the early stages of the fungal development suggested spontaneous posttranslational modification of the native PFK1 in A. niger cells, while from the kinetic parameters determined for both isolated forms it could be concluded that the fragmented enzyme might be more efficient under physiological conditions. PMID- 15746346 TI - Quantitative molecular assay for fingerprinting microbial communities of wastewater and estrogen-degrading consortia. AB - A quantitative fingerprinting method, called the real-time terminal restriction fragment length polymorphism (real-time-t-RFLP) assay, was developed for simultaneous determination of microbial diversity and abundance within a complex community. The real-time-t-RFLP assay was developed by incorporating the quantitative feature of real-time PCR and the fingerprinting feature of t-RFLP analysis. The assay was validated by using a model microbial community containing three pure strains, an Escherichia coli strain (gram negative), a Pseudomonas fluorescens strain (gram negative), and a Bacillus thuringiensis strain (gram positive). Subsequently, the real-time-t-RFLP assay was applied to and proven to be useful for environmental samples; the richness and abundance of species in microbial communities (expressed as the number of 16S rRNA gene copies of each ribotype per milliliter) of wastewater and estrogen-degrading consortia (enriched with 17alpha-estradiol, 17beta-estradiol, or estrone) were successfully characterized. The results of this study strongly suggested that the real-time-t RFLP assay can be a powerful molecular tool for gaining insight into microbial communities in various engineered systems and natural habitats. PMID- 15746347 TI - Molecular and conformational basis of a specific and high-affinity interaction between AlbA and albicidin phytotoxin. AB - The albA gene of Klebsiella oxytoca encodes a protein of 221 amino acids that binds the albicidin phytotoxin with a high affinity (dissociation constant = 6.4 x 10(-8) M). For this study, circular dichroism (CD) spectrometry and an alanine scanning mutagenesis approach were used in combination to investigate the molecular and conformational mechanisms of this high-affinity protein-ligand interaction. CD analysis revealed that AlbA contains a high-affinity binding site, and binding of the albicidin ligand to AlbA in a low-ionic-strength environment induced significant conformational changes. The ligand-dependent conformational changes of AlbA were specific and rapid and reached a stable plateau within seconds after the addition of the antibiotic. However, such conformational changes were not detected when AlbA and albicidin were mixed in the high-ionic-strength buffer that is required for maximal binding activity. Based on the conceptual model of protein-ligand interaction, we propose that a threshold ion strength allows AlbA to complete its conformational rearrangement and resume its original stable structure for accommodation of the bound albicidin. Mutagenesis analysis showed that the replacement of Lys106, Trp110, Tyr113, Leu114, Tyr126, Pro134, and Trp162 with alanine did not change the overall conformational structure of AlbA but decreased the albicidin binding activity about 30 to 60%. We conclude that these residues, together with the previously identified essential residue His125, constitute a high-affinity binding pocket for the ligand albicidin. The results also suggest that hydrophobic and electrostatic potentials of these key amino acid residues may play important roles in the AlbA-albicidin interaction. PMID- 15746349 TI - Functional characterization of two cytochrome P450 monooxygenase genes, P450-1 and P450-4, of the gibberellic acid gene cluster in Fusarium proliferatum (Gibberella fujikuroi MP-D). AB - Gibberella fujikuroi is a species complex with at least nine different biological species, termed mating populations (MPs) A to I (MP-A to MP-I), known to produce many different secondary metabolites. So far, gibberellin (GA) production is restricted to Fusarium fujikuroi (G. fujikuroi MP-C), although at least five other MPs contain all biosynthetic genes. Here, we analyze the GA gene cluster and GA pathway in the closest related species, Fusarium proliferatum (MP-D), and demonstrate that the GA genes share a high degree of sequence homology with the corresponding genes of MP-C. The GA production capacity was restored after integration of the entire GA gene cluster from MP-C, indicating the existence of an active regulation system in F. proliferatum. The results further indicate that one reason for the loss of GA production is the accumulation of several mutations in the coding and 5' noncoding regions of the ent-kaurene oxidase gene, P450-4. PMID- 15746348 TI - Presence of noroviruses and other enteric viruses in sewage and surface waters in The Netherlands. AB - Since virus concentrations in drinking waters are generally below the detection limit, the infectious risk from drinking water consumption requires assessment from the virus concentrations in source waters and removal efficiency of treatment processes. In this study, we estimated from reverse transcription-PCR on 10-fold serially diluted RNA that noroviruses, the most prevalent waterborne gastroenteritis agents, were present at 4 (0.2 to 38) to 4,900 (303 to 4.6 x 10(4)) PCR-detectable units (PDU) per liter of river water (ranges are given in parentheses). These virus concentrations are still high compared with 896 to 7,499 PDU/liter of treated sewage and 5,111 to 850,000 PDU/liter in raw sewage. Sequencing analyses designated human norovirus GGII.4 Lordsdale as the most prevalent strain in the sampling period 1998 to 1999 in both sewage and surface waters. Other GGII strains were also very abundant, indicating that the majority of the virus contamination was derived from urban sewage, although very divergent strains and one animal strain were also detected in the surface and sewage waters. Rotaviruses were also detected in two large rivers (the Maas and the Waal) at 57 to 5,386 PDU/liter. The high virus concentrations determined by PCR may in part be explained by the detection of virus RNA instead of infectious particles. Indeed, reoviruses and enteroviruses that can be cultured were present at much lower levels, of 0.3 to 1 and 2 to 10 PFU/liter, respectively. Assuming 1% of the noroviruses and rotaviruses to be infectious, a much higher disease burden than for other viruses can be expected, not only because of the higher levels but also because of these viruses' higher infectivity and attack rates. PMID- 15746350 TI - "Endomicrobia": cytoplasmic symbionts of termite gut protozoa form a separate phylum of prokaryotes. AB - Lignocellulose digestion by wood-feeding termites depends on the mutualistic interaction of unusual, flagellate protists located in their hindgut. Most of the flagellates harbor numerous prokaryotic endosymbionts of so-far-unknown identity and function. Using a full-cycle molecular approach, we show here that the endosymbionts of the larger gut flagellates of Reticulitermes santonensis belong to the so-called termite group 1 (TG-1) bacteria, a group of clones previously obtained exclusively from gut homogenates of Reticulitermes speratus that are only distantly related to other bacteria and are considered a novel bacterial phylum based on their 16S rRNA gene sequences. Fluorescence in situ hybridization with specifically designed oligonucleotide probes confirmed that TG-1 bacteria are indeed located within the flagellate cells and demonstrated that Trichonympha agilis (Hypermastigida) and Pyrsonympha vertens (Oxymonadida) harbor phylogenetically distinct populations of symbionts (<95% sequence similarity). Transmission electron microscopy revealed that the symbionts are small, spindle shaped cells (0.6 microm in length and 0.3 microm in diameter) surrounded by two membranes and located within the cytoplasm of their hosts. The symbionts of the two flagellates are described as candidate species in the candidate genus "Endomicrobium." Moreover, we provide evidence that the members of the TG-1 phylum, for which we propose the candidate name "Endomicrobia," are phylogenetically extremely diverse and are present in and also restricted to the guts of all lower termites and wood-feeding cockroaches of the genus Cryptocercus, the only insects that are in an exclusive, obligately mutualistic association with such unique cellulose-fermenting protists. PMID- 15746351 TI - Incidence and diversity of potentially highly heat-resistant spores isolated at dairy farms. AB - The presence of highly heat-resistant spores of Bacillus sporothermodurans in ultrahigh-temperature or sterilized consumer milk has emerged as an important item in the dairy industry. Their presence is considered undesirable since they hamper the achievement of commercial sterility requirements. By using a selective 30-min heat treatment at 100 degrees C, 17 Belgian dairy farms were screened to evaluate the presence, sources, and nature of potentially highly heat-resistant spores in raw milk. High numbers of these spores were detected in the filter cloth of the milking equipment and in green crop and fodder samples. About 700 strains were isolated after the selective heating, of which 635 could be screened by fatty acid methyl ester analysis. Representative strains were subjected to amplified ribosomal DNA restriction analysis, 16S rRNA gene sequencing, percent G+C content, and DNA-DNA reassociations for further identification. The strain collection showed a remarkable diversity, with representatives of seven aerobic spore-forming genera. Bacillus licheniformis and Bacillus pallidus were the most predominant species overall. Twenty-three percent of the 603 spore-forming isolates proved to belong to 18 separate novel species. These findings suggest that the selective heating revealed a pool of unknown organisms with a higher heat-resistant character. This study showed that high spore counts can occur at the dairy farm and that feed and milking equipment can act as reservoirs or entry points for potentially highly heat-resistant spores into raw milk. Lowering this spore load by good hygienic measures could probably further reduce the contamination level of raw milk, in this way minimizing the aerobic spore-forming bacteria that could lead to spoilage of milk and dairy products. Assessment and characterization of this particular flora are of great importance to allow the dairy or food industry to adequately deal with newly arising microbiological problems. PMID- 15746352 TI - Quantitative-PCR assessment of Cryptosporidium parvum cell culture infection. AB - A quantitative TaqMan PCR method was developed for assessing the Cryptosporidium parvum infection of in vitro cultivated human ileocecal adenocarcinoma (HCT-8) cell cultures. This method, termed cell culture quantitative sequence detection (CC-QSD), has numerous applications, several of which are presented. CC-QSD was used to investigate parasite infection in cell culture over time, the effects of oocyst treatment on infectivity and infectivity assessment of different C. parvum isolates. CC-QSD revealed that cell culture infection at 24 and 48 h postinoculation was approximately 20 and 60%, respectively, of the endpoint 72-h postinoculation infection. Evaluation of three different lots of C. parvum Iowa isolate oocysts revealed that the mean infection of 0.1 N HCl-treated oocysts was only 36% of the infection obtained with oocysts treated with acidified Hanks' balanced salt solution containing 1% trypsin. CC-QSD comparison of the C. parvum Iowa and TAMU isolates revealed significantly higher levels of infection for the TAMU isolate, which agrees with and supports previous human, animal, and cell culture studies. CC-QSD has the potential to aid in the optimization of Cryptosporidium cell culture methods and facilitate quantitative evaluation of cell culture infectivity experiments. PMID- 15746353 TI - Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. AB - Although copious qualitative information describes the members of the diverse microbial communities on Earth, statistical approaches for quantifying and comparing the numbers and compositions of lineages in communities are lacking. We present a method that addresses the challenge of assigning sequences to operational taxonomic units (OTUs) based on the genetic distances between sequences. We developed a computer program, DOTUR, which assigns sequences to OTUs by using either the furthest, average, or nearest neighbor algorithm for each distance level. DOTUR uses the frequency at which each OTU is observed to construct rarefaction and collector's curves for various measures of richness and diversity. We analyzed 16S rRNA gene libraries derived from Scottish and Amazonian soils and the Sargasso Sea with DOTUR, which assigned sequences to OTUs rapidly and reliably based on the genetic distances between sequences and identified previous inconsistencies and errors in assigning sequences to OTUs. An analysis of the two 16S rRNA gene libraries from soil demonstrated that they do not contain enough sequences to support a claim that they contain different numbers of bacterial lineages with statistical confidence (P > 0.05), nor do they contain enough sequences to provide a robust estimate of species richness when an OTU is defined as containing sequences that are no more than 3% different from each other. In contrast, the richness of OTUs at the 3% level in the Sargasso Sea collection began to plateau after the sampling of 690 sequences. We anticipate that an equivalent extent of sampling for soil would require sampling more than 10,000 sequences, almost 100 times the size of typical sequence collections obtained from soil. PMID- 15746354 TI - Overproduction of heterologous mannitol 1-phosphatase: a key factor for engineering mannitol production by Lactococcus lactis. AB - To achieve high mannitol production by Lactococcus lactis, the mannitol 1 phosphatase gene of Eimeria tenella and the mannitol 1-phosphate dehydrogenase gene mtlD of Lactobacillus plantarum were cloned in the nisin-dependent L. lactis NICE overexpression system. As predicted by a kinetic L. lactis glycolysis model, increase in mannitol 1-phosphate dehydrogenase and mannitol 1-phosphatase activities resulted in increased mannitol production. Overexpression of both genes in growing cells resulted in glucose-mannitol conversions of 11, 21, and 27% by the L. lactis parental strain, a strain with reduced phosphofructokinase activity, and a lactate dehydrogenase-deficient strain, respectively. Improved induction conditions and increased substrate concentrations resulted in an even higher glucose-to-mannitol conversion of 50% by the lactate dehydrogenase deficient L. lactis strain, close to the theoretical mannitol yield of 67%. Moreover, a clear correlation between mannitol 1-phosphatase activity and mannitol production was shown, demonstrating the usefulness of this metabolic engineering approach. PMID- 15746355 TI - Intracellular pH distribution in Saccharomyces cerevisiae cell populations, analyzed by flow cytometry. AB - Intracellular pH has an important role in the maintenance of the normal functions of yeast cells. The ability of the cell to maintain this pH homeostasis also in response to environmental changes has gained more and more interest in both basic and applied research. In this study we describe a protocol which allows the rapid determination of the intracellular pH of Saccharomyces cerevisiae cells. The method is based on flow cytometry and employs the pH-dependent fluorescent probe carboxy SNARF-4F. The protocol attempts to minimize the perturbation of the system under study, thus leading to accurate information about the physiological state of the single cell. Moreover, statistical analysis performed on major factors that may influence the final determination supported the validity of the optimized protocol. The protocol was used to investigate the effect of external pH on S. cerevisiae cells incubated in buffer. The results obtained showed that stationary cells are better able than exponentially grown cells to maintain their intracellular pH homeostasis independently of external pH changes. Furthermore, analysis of the intracellular pH distribution within the cell populations highlighted the presence of subpopulations characterized by different intracellular pH values. Notably, a different behavior was observed for exponentially grown and stationary cells in terms of the appearance and development of these subpopulations as a response to a changing external pH. PMID- 15746356 TI - In vitro reconstitution of an NADPH-dependent superoxide reduction pathway from Pyrococcus furiosus. AB - A scheme for the detoxification of superoxide in Pyrococcus furiosus has been previously proposed in which superoxide reductase (SOR) reduces (rather than dismutates) superoxide to hydrogen peroxide by using electrons from reduced rubredoxin (Rd). Rd is reduced with electrons from NAD(P)H by the enzyme NAD(P)H:rubredoxin oxidoreductase (NROR). The goal of the present work was to reconstitute this pathway in vitro using recombinant enzymes. While recombinant forms of SOR and Rd are available, the gene encoding P. furiosus NROR (PF1197) was found to be exceedingly toxic to Escherichia coli, and an active recombinant form (rNROR) was obtained via a fusion protein expression system, which produced an inactive form of NROR until cleavage. This allowed the complete pathway from NAD(P)H to the reduction of SOR via NROR and Rd to be reconstituted in vitro using recombinant proteins. rNROR is a 39.9-kDa protein whose sequence contains both flavin adenine dinucleotide (FAD)- and NAD(P)H-binding motifs, and it shares significant similarity with known and putative Rd-dependent oxidoreductases from several anaerobic bacteria, both mesophilic and hyperthermophilic. FAD was shown to be essential for activity in reconstitution assays and could not be replaced by flavin mononucleotide (FMN). The bound FAD has a midpoint potential of -173 mV at 23 degrees C (-193 mV at 80 degrees C). Like native NROR, the recombinant enzyme catalyzed the NADPH-dependent reduction of rubredoxin both at high (80 degrees C) and low (23 degrees C) temperatures, consistent with its proposed role in the superoxide reduction pathway. This is the first demonstration of in vitro superoxide reduction to hydrogen peroxide using NAD(P)H as the electron donor in an SOR-mediated pathway. PMID- 15746357 TI - Two alpha(1-3) glucan synthases with different functions in Aspergillus fumigatus. AB - Alpha(1-3) glucan is a main component of the Aspergillus fumigatus cell wall. In spite of its importance, synthesis of this amorphous polymer has not been investigated to date. Two genes in A. fumigatus, AGS1 and AGS2, are highly homologous to the AGS genes of Schizosaccharomyces pombe, which encode putative alpha(1-3) glucan synthases. The predicted Ags proteins of A. fumigatus have an estimated molecular mass of 270 kDa. AGS1 and AGS2 were disrupted in A. fumigatus. Both Deltaags mutants have similar altered hyphal morphologies and reduced conidiation levels. Only Deltaags1 presented a reduction in the alpha(1 3) glucan content of the cell wall. These results showed that Ags1p and Ags2p were functionally different. The cellular localization of the two proteins was in agreement with their different functions: Ags1p was localized at the periphery of the cell in connection with the cell wall, whereas Ags2p was intracellularly located. An original experimental model of invasive aspergillosis based on mixed infection and quantitative PCR was developed to analyze the virulence of A. fumigatus mutant and wild-type strains. Using this model, it was shown that the cell wall and morphogenesis defects of Deltaags1 and Deltaags2 were not associated with a reduction in virulence in either mutant. This result showed that a 50% reduction in the content of the cell wall alpha(1-3) glucan does not play a significant role in A. fumigatus pathogenicity. PMID- 15746359 TI - In vivo monitoring of obligate biotrophic pathogen growth by kinetic PCR. AB - The plant kingdom is constantly challenged by a battery of evolving pathogens. New species or races of pathogens are discovered on crops that were initially bred for disease resistance, and globalization is facilitating the movement of exotic pests. Among these pests, obligate biotrophic parasites make up some of the most damaging groups and have been particularly challenging to study. Here we demonstrate the utility of kinetic PCR (kPCR) (real-time PCR, quantitative PCR) to assess the growth of poplar rust, caused by Melampsora species, by quantification of pathogen DNA. kPCR allowed the construction of reliable growth curves from inoculation through the final stages of uredinial maturation, as well as pathogen monitoring before symptoms become visible. Growth parameters, such as latency period, generation time in logarithmic growth, and the increase in DNA mass at saturation, were compared in compatible, incompatible, and nonhost interactions. Pathogen growth was monitored in different applications dealing with plant pathology, such as host and pathogen diversity and transgenic crop improvement. Finally, the capacity of kPCR to differentiate pathogens in the same sample has broad molecular ecology applications for dynamically monitoring the growth of fungi in their environments or in mixed populations or to measure the efficacy of pest control strategies. PMID- 15746358 TI - Role of cis-acting sites NorL, a TATA box, and AflR1 in nor-1 transcriptional activation in Aspergillus parasiticus. AB - The transcription factor AflR is required for up-regulation of specific pathway genes involved in aflatoxin biosynthesis in the filamentous fungus Aspergillus. nor-1 encodes an early aflatoxin pathway enzyme; its promoter contains a consensus AflR binding site (AflR1). Proteins in Aspergillus parasiticus cell extracts and AflR expressed in Escherichia coli do not bind to A. parasiticus AflR1 in vitro, so it was not clear if this site was required for nor-1 expression or if other transcription factors contributed to gene regulation. In this study we defined the role of AflR1 in nor-1 expression in A. parasiticus and identified additional cis-acting sites required for maximum nor-1 transcriptional activation. Deletion and substitution of AflR1 in the nor-1 promoter in A. parasiticus nor-1::GUS reporter strains showed that this site is required for nor 1 transcriptional activation in vivo. Substitution of a putative TATA box in the nor-1 promoter resulted in nondetectable beta-glucuronidase (GUS) activity, demonstrating that this TATA box is functional in vivo. We also identified a novel cis-acting site, designated NorL, between residues -210 and -238 that was required for maximum nor-1 transcriptional activation in A. parasiticus grown in liquid medium and on solid medium. Using an electrophoretic mobility shift assay, we identified a specific NorL-dependent DNA-protein complex that relies on a functional AflR, either directly or indirectly, for maximum binding capacity. Because the NorL site appears only once in the aflatoxin gene cluster, its association with the nor-1 promoter may have important implications for the overall regulatory scheme for the aflatoxin pathway. PMID- 15746360 TI - Coexistence of bacterial sulfide oxidizers, sulfate reducers, and spirochetes in a gutless worm (Oligochaeta) from the Peru margin. AB - Olavius crassitunicatus is a small symbiont-bearing worm that occurs at high abundance in oxygen-deficient sediments in the East Pacific Ocean. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization, we examined the diversity and phylogeny of bacterial symbionts in two geographically distant O. crassitunicatus populations (separated by 385 km) on the Peru margin (water depth, approximately 300 m). Five distinct bacterial phylotypes co occurred in all specimens from both sites: two members of the gamma Proteobacteria (Gamma 1 and 2 symbionts), two members of the delta-Proteobacteria (Delta 1 and 2 symbionts), and one spirochete. A sixth phylotype belonging to the delta-Proteobacteria (Delta 3 symbiont) was found in only one of the two host populations. Three of the O. crassitunicatus bacterial phylotypes are closely related to symbionts of other gutless oligochaete species; the Gamma 1 phylotype is closely related to sulfide-oxidizing symbionts of Olavius algarvensis, Olavius loisae, and Inanidrilus leukodermatus, the Delta 1 phylotype is closely related to sulfate-reducing symbionts of O. algarvensis, and the spirochete is closely related to spirochetal symbionts of O. loisae. In contrast, the Gamma 2 phylotype and the Delta 2 and 3 phylotypes belong to novel lineages that are not related to other bacterial symbionts. Such a phylogenetically diverse yet highly specific and stable association in which multiple bacterial phylotypes coexist within a single host has not been described previously for marine invertebrates. PMID- 15746361 TI - Enhanced survival of Salmonella enterica in vesicles released by a soilborne Tetrahymena species. AB - Nondestructive ingestion by soilborne protozoa may enhance the environmental resiliency of important bacterial pathogens and may model how such bacteria evade destruction in human macrophages. Here, the interaction of Salmonella enterica serovar Thompson with a soilborne Tetrahymena sp. isolate was examined using serovar Thompson cells labeled with the green fluorescent protein. The bacteria were mixed in solution with cells of Tetrahymena at several ratios. During incubation with serovar Thompson, Tetrahymena cells released a large number of vesicles containing green fluorescent serovar Thompson cells. In comparison, grazing on Listeria monocytogenes cells resulted in their digestion and thus the infrequent release of this pathogen in vesicles. The number of serovar Thompson cells per vesicle increased significantly as the initial ratio of serovar Thompson to Tetrahymena cells increased from 500:1 to 5,000:1. The density of serovar Thompson was as high as 50 cells per vesicle. Staining with propidium iodide revealed that a significantly higher proportion of serovar Thompson cells remained viable when enclosed in vesicles than when free in solution. Enhanced survival rates were observed in vesicles that were secreted by both starved (F = 28.3, P < 0.001) and unstarved (F = 14.09, P < 0.005) Tetrahymena cells. Sequestration in vesicles also provided greater protection from low concentrations of calcium hypochlorite. Thus, the release of this human pathogen from Tetrahymena cells in high-density clusters enclosed in a membrane may have important implications for public health. PMID- 15746362 TI - Subtle difference between benzene and toluene dioxygenases of Pseudomonas putida. AB - Benzene dioxygenase and toluene dioxygenase from Pseudomonas putida have similar catalytic properties, structures, and gene organizations, but they differ in substrate specificity, with toluene dioxygenase having higher activity toward alkylbenzenes. The catalytic iron-sulfur proteins of these enzymes consist of two dissimilar subunits, alpha and beta; the alpha subunit contains a [2Fe-2S] cluster involved in electron transfer, the catalytic nonheme iron center, and is also responsible for substrate specificity. The amino acid sequences of the alpha subunits of benzene and toluene dioxygenases differ at only 33 of 450 amino acids. Chimeric proteins and mutants of the benzene dioxygenase alpha subunit were constructed to determine which of these residues were primarily responsible for the change in specificity. The protein containing toluene dioxygenase C terminal region residues 281 to 363 showed greater substrate preference for alkyl benzenes. In addition, we identified four amino acid substitutions in this region, I301V, T305S, I307L, and L309V, that particularly enhanced the preference for ethylbenzene. The positions of these amino acids in the alpha subunit structure were modeled by comparison with the crystal structure of naphthalene dioxygenase. They were not in the substrate-binding pocket but were adjacent to residues that lined the channel through which substrates were predicted to enter the active site. However, the quadruple mutant also showed a high uncoupled rate of electron transfer without product formation. Finally, the modified proteins showed altered patterns of products formed from toluene and ethylbenzene, including monohydroxylated side chains. We propose that these properties can be explained by a more facile diffusion of the substrate in and out of the substrate cavity. PMID- 15746363 TI - Characterization of the highly efficient sucrose isomerase from Pantoea dispersa UQ68J and cloning of the sucrose isomerase gene. AB - Sucrose isomerase (SI) genes from Pantoea dispersa UQ68J, Klebsiella planticola UQ14S, and Erwinia rhapontici WAC2928 were cloned and expressed in Escherichia coli. The predicted products of the UQ14S and WAC2928 genes were similar to known SIs. The UQ68J SI differed substantially, and it showed the highest isomaltulose producing efficiency in E. coli cells. The purified recombinant WAC2928 SI was unstable, whereas purified UQ68J and UQ14S SIs were very stable. UQ68J SI activity was optimal at pH 5 and 30 to 35 degrees C, and it produced a high ratio of isomaltulose to trehalulose (>22:1) across its pH and temperature ranges for activity (pH 4 to 7 and 20 to 50 degrees C). In contrast, UQ14S SI showed optimal activity at pH 6 and 35 degrees C and produced a lower ratio of isomaltulose to trehalulose (<8:1) across its pH and temperature ranges for activity. UQ68J SI had much higher catalytic efficiency; the Km was 39.9 mM, the Vmax was 638 U mg( 1), and the Kcat/Km was 1.79 x 10(4) M(-1) s(-1), compared to a Km of 76.0 mM, a Vmax of 423 U mg(-1), and a Kcat/Km of 0.62 x 10(4) M(-1) s(-1) for UQ14S SI. UQ68J SI also showed no apparent reverse reaction producing glucose, fructose, or trehalulose from isomaltulose. These properties of the P. dispersa UQ68J enzyme are exceptional among purified SIs, and they indicate likely differences in the mechanism at the enzyme active site. They may favor the production of isomaltulose as an inhibitor of competing microbes in high-sucrose environments, and they are likely to be highly beneficial for industrial production of isomaltulose. PMID- 15746364 TI - Signal transduction by Tga3, a novel G protein alpha subunit of Trichoderma atroviride. AB - Trichoderma species are used commercially as biocontrol agents against a number of phytopathogenic fungi due to their mycoparasitic characterisitics. The mycoparasitic response is induced when Trichoderma specifically recognizes the presence of the host fungus and transduces the host-derived signals to their respective regulatory targets. We made deletion mutants of the tga3 gene of Trichoderma atroviride, which encodes a novel G protein alpha subunit that belongs to subgroup III of fungal Galpha proteins. Deltatga3 mutants had changes in vegetative growth, conidiation, and conidial germination and reduced intracellular cyclic AMP levels. These mutants were avirulent in direct confrontation assays with Rhizoctonia solani or Botrytis cinerea, and mycoparasitism-related infection structures were not formed. When induced with colloidal chitin or N-acetylglucosamine in liquid culture, the mutants had reduced extracellular chitinase activity even though the chitinase-encoding genes ech42 and nag1 were transcribed at a significantly higher rate than they were in the wild type. Addition of exogenous cyclic AMP did not suppress the altered phenotype or restore mycoparasitic overgrowth, although it did restore the ability to produce the infection structures. Thus, T. atroviride Tga3 has a general role in vegetative growth and can alter mycoparasitism-related characteristics, such as infection structure formation and chitinase gene expression. PMID- 15746365 TI - Genomic analysis of bacteriophage PhiJL001: insights into its interaction with a sponge-associated alpha-proteobacterium. AB - Bacteriophage PhiJL001 infects a novel marine bacterium in the alpha subclass of the Proteobacteria isolated from the marine sponge Ircinia strobilina. PhiJL001 is a siphovirus and forms turbid plaques on its host. The genome sequence of PhiJL001 was determined in order to better understand the interaction between the marine phage and its sponge-associated host bacterium. The complete genome sequence of PhiJL001 comprised 63,469 bp with an overall G+C content of 62%. The genome has 91 predicted open reading frames (ORFs), and 17 ORFs have been assigned putative functions. PhiJL001 appears to be a temperate phage, and the integrase gene was identified in the genome. DNA hybridization analysis showed that the PhiJL001 genome does not integrate into the host chromosome under the conditions tested. DNA hybridization experiments therefore suggested that PhiJL001 has some pseudolysogenic characteristics. The genome of PhiJL001 contains many putative genes involved in phage DNA replication (e.g., helicase, DNA polymerase, and thymidylate synthase genes) and also contains a putative integrase gene associated with the lysogenic cycle. Phylogeny based on DNA polymerase gene sequences indicates that PhiJL001 is related to a group of siphoviruses that infect mycobacteria. Designation of PhiJL001 as a siphovirus is consistent with the morphology of the phage visualized by transmission electron microscopy. The unique marine phage-host system described here provides a model system for studying the role of phages in sponge microbial communities. PMID- 15746366 TI - Natural dissemination of Bacillus anthracis spores in northern Canada. AB - Soil samples were collected from around fresh and year-old bison carcasses and areas not associated with known carcasses in Wood Buffalo National Park during an active anthrax outbreak in the summer of 2001. Sample selection with a grid provided the most complete coverage of a site. Soil samples were screened for viable Bacillus anthracis spores via selective culture, phenotypic analysis, and PCR. Bacillus anthracis spores were isolated from 28.4% of the samples. The highest concentrations of B. anthracis spores were found directly adjacent to fresh carcasses and invariably corresponded to locations where the soil had been saturated with body fluids escaping the carcass through either natural body orifices or holes torn by scavengers. The majority of positive samples were found within 2 m of both year-old and fresh carcasses and probably originated from scavengers churning up and spreading the body fluid-saturated soil as they fed. Trails of lesser contamination radiating from the carcasses probably resulted from spore dissemination through adhesion to scavengers and through larger scavengers dragging away disarticulated limbs. Comparison of samples from minimally scavenged and fully necropsied carcass sites revealed no statistically significant difference in the level of B. anthracis spore contamination. Therefore, the immediate area around a suspected anthrax carcass should be considered substantially contaminated regardless of the condition of the carcass. PMID- 15746367 TI - Genomic comparisons of Salmonella enterica serovar Dublin, Agona, and Typhimurium strains recently isolated from milk filters and bovine samples from Ireland, using a Salmonella microarray. AB - Salmonella-induced enterocolitis is the leading food-borne illness with a lethal outcome and causes millions of cases of gastroenteritis each year. We examined genomic variation among 12 environmental, veterinary, and clinical Salmonella enterica serovar Dublin, Agona, and Typhimurium strains isolated in Ireland between 2000 and 2003, as well as two clinical isolates from Canada and four archival isolates, which belonged to serovars Dublin and Agona. Using DNA-DNA hybridization to a microarray consisting of most of the predicted protein encoding sequences of the S. enterica serovar Typhimurium LT2 genome, we identified a number of genomic regions that were absent in one or more serovars. The 34 genomic regions encoded proteins involved in sugar metabolism, transport, fimbrial and phage biogenesis, and transcriptional regulation, as well as inner and outer membrane-associated proteins. Two of the four prophages identified in strain LT2, prophages Gifsy-1 and Gifsy-2, were present in all six serovar Typhimurium strains examined. Prophage Fels-1 was absent from all 18 isolates examined, and Fels-2 was completely absent from the serovar Typhimurium isolates and the Salmonella Reference Collection B serovar Dublin strain Du2. All five Salmonella pathogenicity islands were present in all isolates. Plasmid pSLT was absent from all serovar Agona isolates, and only homologues of the spv genes were present in eight of the nine serovar Dublin strains. Only limited intraserovar diversity was found among the nine serovar Dublin, three serovar Agona, and six serovar Typhimurium isolates examined even though these isolates had extensive geographic, temporal, and source differences. PMID- 15746368 TI - Chemotaxonomic identification of single bacteria by micro-Raman spectroscopy: application to clean-room-relevant biological contaminations. AB - Microorganisms, such as bacteria, which might be present as contamination inside an industrial food or pharmaceutical clean room process need to be identified on short time scales in order to minimize possible health hazards as well as production downtimes causing financial deficits. Here we describe the first results of single-particle micro-Raman measurements in combination with a classification method, the so-called support vector machine technique, allowing for a fast, reliable, and nondestructive online identification method for single bacteria. PMID- 15746369 TI - Environmental DNA fragment conferring early and increased sporulation and antibiotic production in Streptomyces species. AB - Here we describe the rep gene, isolated from an environmental DNA library, which when transformed into Streptomyces species resulted in increased production of secondary metabolites and accelerated sporulation. We show that Streptomyces lividans strains bearing rep are particularly useful as expression hosts for heterologous antibiotic production. PMID- 15746370 TI - Heterologous expression, purification, and characterization of a highly active xylose reductase from Neurospora crassa. AB - A xylose reductase (XR) gene was identified from the Neurospora crassa whole genome sequence, expressed heterologously in Escherichia coli, and purified as a His6-tagged fusion in high yield. This enzyme is one of the most active XRs thus far characterized and may be used for the in vitro production of xylitol. PMID- 15746371 TI - Comparison of diversities of Escherichia coli O157 shed from a cohort of spring born beef calves at pasture and in housing. AB - A cohort of spring-born beef calves demonstrated limited genetic and phenotypic diversity of Escherichia coli O157 when kept in a state of isolation. Despite this, there was a difference in the pulsed-field gel electrophoresis and phage types of isolates shed by cattle at pasture compared with those shed by the same cattle when weaned and housed. PMID- 15746372 TI - Effect of batch-process solar disinfection on survival of Cryptosporidium parvum oocysts in drinking water. AB - The results of batch-process solar disinfection (SODIS) of Cryptosporidium parvum oocysts in water are reported. Oocyst suspensions were exposed to simulated sunlight (830 W m(-2)) at 40 degrees C. Viability assays (4',6'-diamidino-2 phenylindole [DAPI]/propidium iodide and excystation) and infectivity tests (Swiss CD-1 suckling mice) were performed. SODIS exposures of 6 and 12 h reduced oocyst infectivity from 100% to 7.5% (standard deviation = 2.3) and 0% (standard deviation = 0.0), respectively. PMID- 15746373 TI - Genetic diversity in the lactose operons of Lactobacillus helveticus strains and its relationship to the role of these strains as commercial starter cultures. AB - Two novel insertion sequence elements, ISLhe1 and ISLhe15, were located upstream of the genes encoding the beta-galactosidase enzyme in Lactobacillus helveticus commercial starter strains. Strains with the IS982 family element, ISLhe1, demonstrated reduced beta-galactosidase activity compared to the L. helveticus type strain, whereas strains with the ISLhe15 element expressed beta galactosidase in the absence of lactose. PMID- 15746374 TI - Dynamics of ribulose 1,5-bisphosphate carboxylase/oxygenase gene expression in the coccolithophorid Coccolithus pelagicus during a tracer release experiment in the Northeast Atlantic. AB - We report a pronounced diel rhythm in ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) gene expression in a natural population of the coccolithophorid Coccolithus pelagicus sampled during a Lagrangian experiment in the Northeast Atlantic. Our observations show that there is greater heterogeneity in the temporal regulation of RubisCO expression among planktonic chromophytes than has been reported hitherto. PMID- 15746375 TI - Probiotic bacteria may become dormant during storage. AB - The determination of bacterial viability in probiotic products is of economic, technological, and clinical significance. We compared four methods to enumerate three Bifidobacterium strains in fermented oat products during storage. A subpopulation of nonculturable cells retained a functional cell membrane typical of viable cells, indicating that probiotic bacteria become dormant during storage. PMID- 15746376 TI - Characterization of hydrogenase and reductive dehalogenase activities of Dehalococcoides ethenogenes strain 195. AB - Dehalococcoides ethenogenes strain 195 reductively dechlorinates tetrachloroethene (PCE) and trichloroethene (TCE) to vinyl chloride and ethene using H2 as an electron donor. PCE- and TCE-reductive dehalogenase (RD) activities were mainly membrane associated, whereas only about 20% of the hydrogenase activity was membrane associated. Experiments with methyl viologen (MV) were consistent with a periplasmic location for the RDs or a component feeding electrons to them. The protonophore uncoupler tetrachlorosalicylanilide did not inhibit reductive dechlorination in cells incubated with H2 and PCE and partially restored activity in cells incubated with the ATPase inhibitor N,N' dicyclohexylcarbodiimide. Benzyl viologen or diquat (Eo' approximately -360 mV) supported reductive dechlorination of PCE or TCE at rates comparable to MV (-450 mV) in cell extracts. PMID- 15746377 TI - Optical brighteners do not influence covert baculovirus infection of Spodoptera frugiperda. AB - Covert infection with Spodoptera frugiperda multiple nucleopolyhedrovirus, detected by reverse transcription-PCR of virus gene transcripts (ie-0 and polh), was not significantly affected by the presence of an optical brightener (Tinopal UNPA-GX), indicating no change in virus virulence. Detection of the covert infection was dependent on insect life stage and the viral mRNA used for diagnosis. PMID- 15746379 TI - Calcium dysregulation, IP3 signaling, and Alzheimer's disease. AB - Ca(2+) ions subserve complex signaling roles in neurons, regulating functions ranging from gene transcription to modulation of membrane excitability. Ca(2+) ions enter the cytosol from extracellular sources, such as entry through voltage gated channels, and by liberation from intracellular endoplasmic reticulum (ER) stores through inositol triphosphate (IP(3)) receptors and/or ryanodine (RyR) receptors. Disruptions of intracellular Ca(2+) signaling are proposed to underlie the pathophysiology of Alzheimer's disease (AD), and recent studies examining AD linked mutations in the presenilin genes demonstrate enhanced ER Ca(2+) release in a variety of cell types and model systems. The development of transgenic AD mouse models provides a means to study the mechanisms and downstream effects of neuronal ER Ca(2+)-signaling alterations on AD pathogenesis and offers insight into potential novel therapeutic strategies. The author discusses recent findings in both the physiological functioning of the IP(3)-signaling pathway in neurons and the involvement of ERCa(2+) disruptions in the pathogenesis of AD. PMID- 15746378 TI - Application of a newly developed ARB software-integrated tool for in silico terminal restriction fragment length polymorphism analysis reveals the dominance of a novel pmoA cluster in a forest soil. AB - TRF-CUT, an ARB-implemented tool, was developed to predict in silico the terminal restriction fragments of aligned small-subunit rRNA gene or functional gene sequences. Application of this new tool to perform directed terminal restriction fragment length polymorphism analysis of pmoA products obtained from a forest soil revealed that novel cluster I methanotrophic bacteria were dominant. PMID- 15746380 TI - TRPM7 and ischemic CNS injury. AB - Ischemic brain damage represents a major source of morbidity and mortality in westernized society and poses a significant financial burden on the health care system. To date, few effective therapies have been realized to treat stroke and once promising avenues such as antiexcitotoxic therapy with NMDA receptor antagonists have not proven clinically useful. Thus, we need to identify new targets for research and therapeutic intervention of the neurodegeneration caused by stroke. Transient receptor potential (TRP) channels are an exciting new family of cation channels that respond to intracellular and extracellular stimuli. Indeed, several members can be induced by oxidative stress and oxygen free radicals. We have recently demonstrated that one member, TRPM7, is an essential mediator of anoxic neuronal death that is activated by oxidative stress, in parallel to excitotoxic signal pathways. Thus, future treatment of ischemic brain injury may need to include strategies that inhibit or modulate TRPM7 activity. Further investigation of the physiology and pathophysiology of TRPM7 and other TRP family members is needed to provide both pharmacological targets and a better understanding of ischemic brain disorders. PMID- 15746381 TI - The control of voluntary eye movements: new perspectives. AB - Primates use two types of voluntary eye movements to track objects of interest: pursuit and saccades. Traditionally, these two eye movements have been viewed as distinct systems that are driven automatically by low-level visual inputs. However, two sets of findings argue for a new perspective on the control of voluntary eye movements. First, recent experiments have shown that pursuit and saccades are not controlled by entirely different neural pathways but are controlled by similar networks of cortical and subcortical regions and, in some cases, by the same neurons. Second, pursuit and saccades are not automatic responses to retinal inputs but are regulated by a process of target selection that involves a basic form of decision making. The selection process itself is guided by a variety of complex processes, including attention, perception, memory, and expectation. Together, these findings indicate that pursuit and saccades share a similar functional architecture. These points of similarity may hold the key for understanding how neural circuits negotiate the links between the many higher order functions that can influence behavior and the singular and coordinated motor actions that follow. PMID- 15746382 TI - Exocytosis and endocytosis of synaptic vesicles and functional roles of vesicle pools: lessons from the Drosophila neuromuscular junction. AB - To maintain synaptic transmission during intense neuronal activities, the synaptic vesicle (SV) pool at release sites is effectively replenished by recruitment of SVs from the reserve pool and/or by endocytosis. The authors have studied dynamics of SVs using a fluorescence dye, FM1-43, which is incorporated into SVs during endocytosis and released by exocytosis. Drosophila is one of the most suitable preparations for genetic and pharmacological analyses, and this provides a useful model system. The authors found at the neuromuscular junctions of Drosophila that exocytosis and endocytosis of SVs are triggered by Ca(2+) influx through distinct routes and that selective inhibition of exocytosis or endocytosis resulted in depression of synaptic transmission with a distinct time course. They identified two SV pools in a single presynaptic bouton. The exo/endo cycling pool (ECP) is loaded with FM1-43 during low-frequency stimulation and locates close to release sites in the periphery of boutons, whereas the reserve pool (RP) is loaded and unloaded only during high-frequency stimulation and resides primarily in the center of boutons. The size of ECP closely correlates with the quantal content of evoked release, suggesting that SVs in the ECP are primarily involved in synaptic transmission. SVs in the RP are recruited to synaptic transmission by a process involving the cAMP/PKA cascade during high frequency stimulation. Cytochalasin D blocked this recruitment process, suggesting involvement of filamentous actin. Endocytosed SVs replenish the ECP during stimulation and the RP after tetanic stimulation. Replenishment of the ECP depends on Ca(2+) influx from external solutions, and that of the RP is initiated by Ca(2+) release from internal stores. Thus, SV dynamics is closely involved in modulation of synaptic efficacy and influences synaptic plasticity. PMID- 15746383 TI - Kalirin: a dual Rho guanine nucleotide exchange factor that is so much more than the sum of its many parts. AB - A large number of Rho guanine nucleotide exchange factors (GEFs) and Rho GTPase activating proteins (GAPs) are used in the CNS to activate specific Rho GTPase family members, thereby inducing various signaling mechanisms that regulate neuronal shape, growth, and plasticity, in part through their effects on the actin cytoskeleton. Kalirin is a large neuronal dual Rho GEF that activates Rac1, RhoA, and RhoG via its two Rho GEF domains. This activation, which is spatially and temporally regulated, allows Kalirin to influence neurite initiation, axonal growth, and dendritic morphogenesis. In addition, this alternatively spliced gene generates developmentally regulated transcripts that yield proteins localized to the postsynaptic density (PSD). Kalirin-7, which interacts with PSD-95, is necessary for dendritic spine formation. In addition, Kalirins have the ability to regulate and influence other aspects of neuronal morphogenesis via protein protein interactions with their other domains, including many spectrins, other protein and lipid interaction domains, and a potential kinase. These interactions have implications not only for neuronal morphogenesis but also for vesicle trafficking, secretion, neuronal maintenance, and neurodegenerative disease. PMID- 15746384 TI - The corticospinal system: from development to motor control. AB - The corticospinal system is the principal motor system for controlling movements that require the greatest skill and flexibility. It is the last motor system to develop. The pattern of termination of corticospinal axons, as they grow into the spinal gray matter, bears little resemblance to the pattern later in development and in maturity. Refinement of corticospinal terminations occurs during a protracted postnatal period and includes both elimination of transient terminations and growth to new targets. This refinement is driven by neural activity in the motor cortical areas and by limb motor experience. Developing corticospinal terminals compete with each other for synaptic space on spinal neurons. More active terminals are more competitive and are able to secure more synaptic space than their less active counterparts. Corticospinal terminals can activate spinal neurons from very early in development. The importance of this early synaptic activity appears to be more for refining corticospinal connections than for transmitting signals to spinal motor circuits for movement control. The motor control functions of the corticospinal system are not expressed until development of connectional specificity with spinal cord neurons, a strong capacity for corticospinal synapses to facilitate spinal motor circuits, and the formation of the cortical motor map. PMID- 15746385 TI - Adherence to hepatitis B virus vaccination at syringe exchange sites. AB - Injection drug users (IDUs) are at high risk for hepatitis B virus (HBV); however, they often do not receive preventive vaccination. IDUs who use mobile health care services linked to a syringe exchange program in New Haven were routinely screened for HBV, hepatitis C virus, and syphilis. Individuals without prior exposure to HBV were offered three-part vaccination series. Of the 212 IDUs screened for HBV infection, 134 (63%) were eligible (negative for HBV surface and core anti-bodies) for vaccination and 10 (4.7%) had evidence of chronic HBV infection. Compared to those with previous exposure to HBV, vaccine-eligible patients were significantly more likely to be younger and use heroin and less likely to be black, home-less, daily injectors, and cocaine users. Of the 134 vaccine-eligible subjects, 103 (77%) and 89 (66%) completed two and three vaccinations, respectively. Correlates of completing all three vaccinations included older age (OR = 1.06, 95% CI = 1.04-1.07), injecting daily (OR = 2.12, 95% CI = 1.36-6.73), and being homeless (OR = 1.98, 95% CI = 1.14-12.27). These results suggest that IDUs remain at high risk for acquiring HBV infection. Programs that link health care to a syringe exchange program are effective ways to provide preventive health care services to IDUs, particularly HBV vaccination. Trust engendered by and mutual respect afforded by such programs result in repeated encounters by active IDUs over time. PMID- 15746387 TI - Supra-canonical 26Al/27Al and the residence time of CAIs in the solar protoplanetary disk. AB - The canonical initial 26Al/27Al ratio of 4.5 x 10(-5) has been a fiducial marker for the beginning of the solar system. Laser ablation and whole-rock multiple collector inductively coupled plasma-source mass spectrometry magnesium isotope analyses of calcium- and aluminum-rich inclusions (CAIs) from CV3 meteorites demonstrate that some CAIs had initial 26Al/27Al values at least 25% greater than canonical and that the canonical initial 26Al/27Al cannot mark the beginning of solar system formation. Using rates of Mg diffusion in minerals, we find that the canonical initial 26Al/27Al is instead the culmination of thousands of brief high temperature events incurred by CAIs during a 10(5)-year residence time in the solar protoplanetary disk. PMID- 15746386 TI - Activation of a phytopathogenic bacterial effector protein by a eukaryotic cyclophilin. AB - Innate immunity in higher plants invokes a sophisticated surveillance system capable of recognizing bacterial effector proteins. In Arabidopsis, resistance to infection by strains of Pseudomonas syringae expressing the effector AvrRpt2 requires the plant resistance protein RPS2. AvrRpt2 was identified as a putative cysteine protease that results in the elimination of the Arabidopsis protein RIN4. RIN4 cleavage serves as a signal to activate RPS2-mediated resistance. AvrRpt2 is delivered into the plant cell, where it is activated by a eukaryotic factor that we identify as cyclophilin. This activation of AvrRpt2 is necessary for protease activity. Active AvrRpt2 can then directly cleave RIN4. PMID- 15746388 TI - Extracting a climate signal from 169 glacier records. AB - I constructed a temperature history for different parts of the world from 169 glacier length records. Using a first-order theory of glacier dynamics, I related changes in glacier length to changes in temperature. The derived temperature histories are fully independent of proxy and instrumental data used in earlier reconstructions. Moderate global warming started in the middle of the 19th century. The reconstructed warming in the first half of the 20th century is 0.5 kelvin. This warming was notably coherent over the globe. The warming signals from glaciers at low and high elevations appear to be very similar. PMID- 15746390 TI - Bayh-Dole: almost 25. PMID- 15746391 TI - Paleoanthropology. Small but smart? Flores hominid shows signs of advanced brain. PMID- 15746389 TI - Postsynaptic receptor trafficking underlying a form of associative learning. AB - To elucidate molecular, cellular, and circuit changes that occur in the brain during learning, we investigated the role of a glutamate receptor subtype in fear conditioning. In this form of learning, animals associate two stimuli, such as a tone and a shock. Here we report that fear conditioning drives AMPA-type glutamate receptors into the synapse of a large fraction of postsynaptic neurons in the lateral amygdala, a brain structure essential for this learning process. Furthermore, memory was reduced if AMPA receptor synaptic incorporation was blocked in as few as 10 to 20% of lateral amygdala neurons. Thus, the encoding of memories in the lateral amygdala is mediated by AMPA receptor trafficking, is widely distributed, and displays little redundancy. PMID- 15746392 TI - Paleoanthropology. "Hobbit" bones go home to Jakarta. PMID- 15746393 TI - Planetary science. A strange little saturnian ice ball gets stranger still. PMID- 15746394 TI - Computer security. Flaw found in data-protection method. PMID- 15746395 TI - Planetary science. Ice or lava sea on Mars? A transatlantic debate erupts. PMID- 15746396 TI - Conflicts of interest. NIH scientists raise fuss about scope of new rules. PMID- 15746397 TI - French science. Report puts Pasteur move on hold. PMID- 15746398 TI - Infectious diseases. Experts dismiss pig flu scare as nonsense. PMID- 15746399 TI - Canada. Grants councils say more isn't nearly enough to keep science healthy. PMID- 15746400 TI - Human embryonic stem cells. Getting the mice out of ES cell cultures. PMID- 15746401 TI - Retrovirus meeting. Gut assumes sinister new role in HIV pathogenesis. PMID- 15746402 TI - Biodefense. Has biodefense gone overboard? PMID- 15746403 TI - Biodefense. Microbiologist on a mission. PMID- 15746405 TI - U.S. polar research. Shift in icebreaking fleet could crunch NSF budget. PMID- 15746404 TI - Conservation science. What's in a species' name? More than $450,000. PMID- 15746407 TI - Mathematics. Have a Coq and a smile. PMID- 15746406 TI - Mathematics. What in the name of Euclid is going on here? PMID- 15746408 TI - Retraction. PMID- 15746409 TI - An open letter to Elias Zerhouni. PMID- 15746410 TI - A small-scale foreign aid strategy. PMID- 15746411 TI - An explanation for the placebo effect? PMID- 15746412 TI - Comment on "Avian extinction and mammalian introductions on oceanic islands". PMID- 15746414 TI - Materials science. A window on biomineralization. PMID- 15746415 TI - Ocean science. Lost City life. PMID- 15746417 TI - Astronomy. Our interstellar neighborhood. PMID- 15746416 TI - HIV/AIDS. HIV: experiencing the pressures of modern life. PMID- 15746418 TI - Structural biology. Membrane protein insertion and stability. PMID- 15746420 TI - The geometric distance and proper motion of the Triangulum Galaxy (M33). AB - We measured the angular rotation and proper motion of the Triangulum Galaxy (M33) with the Very Long Baseline Array by observing two H2O masers on opposite sides of the galaxy. By comparing the angular rotation rate with the inclination and rotation speed, we obtained a distance of 730 +/- 168 kiloparsecs. This distance is consistent with the most recent Cepheid distance measurement. M33 is moving with a velocity of 190 +/- 59 kilometers per second relative to the Milky Way. These measurements promise a method to determine dynamical models for the Local Group and the mass and dark-matter halos of M31, M33, and the Milky Way. PMID- 15746419 TI - A serpentinite-hosted ecosystem: the Lost City hydrothermal field. AB - The serpentinite-hosted Lost City hydrothermal field is a remarkable submarine ecosystem in which geological, chemical, and biological processes are intimately interlinked. Reactions between seawater and upper mantle peridotite produce methane- and hydrogen-rich fluids, with temperatures ranging from <40 degrees to 90 degrees C at pH 9 to 11, and carbonate chimneys 30 to 60 meters tall. A low diversity of microorganisms related to methane-cycling Archaea thrive in the warm porous interiors of the edifices. Macrofaunal communities show a degree of species diversity at least as high as that of black smoker vent sites along the Mid-Atlantic Ridge, but they lack the high biomasses of chemosynthetic organisms that are typical of volcanically driven systems. PMID- 15746421 TI - Deflection of the interstellar neutral hydrogen flow across the heliospheric interface. AB - Using an absorption cell, we measured the Doppler shifts of the interstellar hydrogen resonance glow to show the direction of the neutral hydrogen flow as it enters the inner heliosphere. The neutral hydrogen flow is found to be deflected relative to the helium flow by about 4 degrees . The most likely explanation of this deflection is a distortion of the heliosphere under the action of an ambient interstellar magnetic field. In this case, the helium flow vector and the hydrogen flow vector constrain the direction of the magnetic field and act as an interstellar magnetic compass. PMID- 15746423 TI - Residential biofuels in South Asia: carbonaceous aerosol emissions and climate impacts. AB - High concentrations of pollution particles, including "soot" or black carbon, exist over the Indian Ocean, but their sources and geographical origins are not well understood. We measured emissions from the combustion of biofuels, used widely in south Asia for cooking, and found that large amounts of carbonaceous aerosols are emitted per kilogram of fuel burnt. We calculate that biofuel combustion is the largest source of black carbon emissions in India, and we suggest that its control is central to climate change mitigation in the south Asian region. PMID- 15746422 TI - Supramolecular assembly of amelogenin nanospheres into birefringent microribbons. AB - Although both tooth enamel and bone are composed of organized assemblies of carbonated apatite crystals, enamel is unusual in that it does not contain collagen nor does it remodel. Self-assembly of amelogenin protein into nanospheres has been recognized as a key factor in controlling the oriented and elongated growth of carbonated apatite crystals during dental enamel biomineralization. We report the in vitro formation of birefringent microribbon structures that were generated through the supramolecular assembly of amelogenin nanospheres. These microribbons have diffraction patterns that indicate a periodic structure of crystalline units along the long axis. The growth of apatite crystals orientated along the c axis and parallel to the long axes of the microribbons was observed in vitro. The linear arrays (chains) of nanospheres observed as intermediate states before the microribbon formation give an important indication as to the function of amelogenin in controlling the oriented growth of apatite crystals during enamel mineralization. PMID- 15746424 TI - Nutritional status and diet composition affect the value of diatoms as copepod prey. AB - The role of diatoms as key food for copepods at the base of pelagic food chains has been questioned recently on the grounds of toxicity. We show, using unialgal versus mixed algal diets of different nutritional status (i.e., nitrogen:carbon ratio) fed to Acartia tonsa, that diatoms per se are not toxic but that single diatom diets are inadequate. Additionally, the nutritional state of the phytoplankton has a profound effect on copepod growth and growth efficiency. The ecological significance of laboratory demonstrations of diatom toxicity needs to be reconsidered. PMID- 15746425 TI - Life at depth: Photobacterium profundum genome sequence and expression analysis. AB - Deep-sea life requires adaptation to high pressure, an extreme yet common condition given that oceans cover 70% of Earth's surface and have an average depth of 3800 meters. Survival at such depths requires specific adaptation but, compared with other extreme conditions, high pressure has received little attention. Recently, Photobacterium profundum strain SS9 has been adopted as a model for piezophily. Here we report its genome sequence (6.4 megabase pairs) and transcriptome analysis. The results provide a first glimpse into the molecular basis for life in the largest portion of the biosphere, revealing high metabolic versatility. PMID- 15746426 TI - A functional dosage compensation complex required for male killing in Drosophila. AB - Bacteria that selectively kill males ("male-killers") were first characterized more than 50 years ago in Drosophila and have proved to be common in insects. However, the mechanism by which sex specificity of virulence is achieved has remained unknown. We tested the ability of Spiroplasma poulsonii to kill Drosophila melanogaster males carrying mutations in genes that encode the dosage compensation complex. The bacterium failed to kill males lacking any of the five protein components of the complex. PMID- 15746427 TI - Extensive DNA inversions in the B. fragilis genome control variable gene expression. AB - The obligately anaerobic bacterium Bacteroides fragilis, an opportunistic pathogen and inhabitant of the normal human colonic microbiota, exhibits considerable within-strain phase and antigenic variation of surface components. The complete genome sequence has revealed an unusual breadth (in number and in effect) of DNA inversion events that potentially control expression of many different components, including surface and secreted components, regulatory molecules, and restriction-modification proteins. Invertible promoters of two different types (12 group 1 and 11 group 2) were identified. One group has inversion crossover (fix) sites similar to the hix sites of Salmonella typhimurium. There are also four independent intergenic shufflons that potentially alter the expression and function of varied genes. The composition of the 10 different polysaccharide biosynthesis gene clusters identified (7 with associated invertible promoters) suggests a mechanism of synthesis similar to the O-antigen capsules of Escherichia coli. PMID- 15746428 TI - Requirement for caspase-8 in NF-kappaB activation by antigen receptor. AB - Caspase-8, a proapoptotic protease, has an essential role in lymphocyte activation and protective immunity. We show that caspase-8 deficiency (CED) in humans and mice specifically abolishes activation of the transcription factor nuclear factor kappaB (NF-kappaB) after stimulation through antigen receptors, Fc receptors, or Toll-like receptor 4 in T, B, and natural killer cells. Caspase-8 also causes the alphabeta complex of the inhibitor of NF-kappaB kinase (IKK) to associate with the upstream Bcl10-MALT1 (mucosa-associated lymphatic tissue) adapter complex. Recruitment of the IKKalpha, beta complex, its activation, and the nuclear translocation of NF-kappaB require enzyme activity of full-length caspase-8. These findings thus explain the paradoxical association of defective apoptosis and combined immunodeficiency in human CED. PMID- 15746429 TI - Impaired thermosensation in mice lacking TRPV3, a heat and camphor sensor in the skin. AB - Environmental temperature is thought to be directly sensed by neurons through their projections in the skin. A subset of the mammalian transient receptor potential (TRP) family of ion channels has been implicated in this process. These "thermoTRPs" are activated at distinct temperature thresholds and are typically expressed in sensory neurons. TRPV3 is activated by heat (>33 degrees C) and, unlike most thermoTRPs, is expressed in mouse keratinocytes. We found that TRPV3 null mice have strong deficits in responses to innocuous and noxious heat but not in other sensory modalities; hence, TRPV3 has a specific role in thermosensation. The natural compound camphor, which modulates sensations of warmth in humans, proved to be a specific activator of TRPV3. Camphor activated cultured primary keratinocytes but not sensory neurons, and this activity was abolished in TRPV3 null mice. Therefore, heat-activated receptors in keratinocytes are important for mammalian thermosensation. PMID- 15746430 TI - OSBP is a cholesterol-regulated scaffolding protein in control of ERK 1/2 activation. AB - Oxysterol-binding protein (OSBP) is the founding member of a family of sterol binding proteins implicated in vesicle transport, lipid metabolism, and signal transduction. Here, OSBP was found to function as a cholesterol-binding scaffolding protein coordinating the activity of two phosphatases to control the extracellular signal-regulated kinase (ERK) signaling pathway. Cytosolic OSBP formed a approximately 440-kilodalton oligomer with a member of the PTPPBS family of tyrosine phosphatases, the serine/threonine phosphatase PP2A, and cholesterol. This oligomer had dual specific phosphatase activity for phosphorylated ERK (pERK). When cell cholesterol was lowered, the oligomer disassembled and the level of pERK rose. The oligomer also disassembled when exposed to oxysterols. Increasing the amount of OSBP oligomer rendered cells resistant to the effects of cholesterol depletion and decreased the basal level of pERK. Thus, cholesterol functions through its interaction with OSBP outside of membranes to regulate the assembly of an oligomeric phosphatase that controls a key signaling pathway in the cell. PMID- 15746431 TI - How visual stimuli activate dopaminergic neurons at short latency. AB - Unexpected, biologically salient stimuli elicit a short-latency, phasic response in midbrain dopaminergic (DA) neurons. Although this signal is important for reinforcement learning, the information it conveys to forebrain target structures remains uncertain. One way to decode the phasic DA signal would be to determine the perceptual properties of sensory inputs to DA neurons. After local disinhibition of the superior colliculus in anesthetized rats, DA neurons became visually responsive, whereas disinhibition of the visual cortex was ineffective. As the primary source of visual afferents, the limited processing capacities of the colliculus may constrain the visual information content of phasic DA responses. PMID- 15746432 TI - Lysozyme secretion by submucosal glands protects the airway from bacterial infection. AB - Submucosal glands are abundant (approximately 1 gland/mm2) secretory structures in the tracheobronchial airways of the human lung. Because submucosal glands express antibacterial proteins, it has been proposed that they contribute to lung defense. However, this concept is challenged by the fact that mice do not have submucosal glands in their bronchial airways, yet are quite resistant to bacterial lung infection. The contribution of airway submucosal glands to host defense is also debated as a pathophysiologic component of cystic fibrosis lung disease. Here, we asked whether submucosal glands protect airways against bacterial infection. By comparing tracheal xenograft airways with and without glands, we found that the presence of glands enhanced bacterial killing in vivo and by airway secretions in vitro. Moreover, immunodepletion studies suggested that lysozyme is a major antibacterial component secreted by submucosal glands. These studies provide evidence that submucosal glands are a major source of antibacterials critical for maintaining sterile airways. PMID- 15746434 TI - Autocrine and paracrine regulation of interleukin-8 expression in lung cancer cells. AB - We had previously demonstrated that lung cancer cells, upon contact with macrophages, could be induced to secrete angiogenic factors to promote tumor angiogenesis. In this study, we focused on the paracrine and autocrine regulation of interleukin (IL)-8 expression in sensitized lung cancer cells after interacting with macrophages. We found that the IL-8 mRNA expression in lung cancer cells significantly increased after coculture with phorbol myristate acetate-treated THP-1 cells and human primary lung macrophages. Fresh lung cancer CL1-5 cells cocultured with macrophage-sensitized lung cancer cells still had a 35% of increase in IL-8 mRNA expression. The addition of anti-inflammatory agents pyrrolidine dithiocarbamate, pentoxifylline, aspirin, and dexamethasone could completely suppress the expression of IL-8 mRNA in fresh/sensitized lung cancer cell cocultures. Human recombinant tumor necrosis factor (TNF)-alpha and IL 1alpha could induce IL-8 expression in lung cancer cells in a dose-dependent manner. Neutralization with TNF-alpha and IL-1alpha antibodies in cocultures decreased the levels of IL-8 expression in sensitized lung cancer cells. Nuclear factor-kappaB transcriptional activity was also suppressed by the same antibodies, as confirmed by a reporter gene assay and the electrophoretic mobility shift assay. Our results highly suggest that both autocrine and paracrine regulation are involved in IL-8 expression of lung cancer cells cocultured with macrophage. Also, the regulations of IL-8 expression in lung cancer cells were through the nuclear factor-kappaB pathway and modulated by TNF alpha and IL-1alpha. PMID- 15746433 TI - Pneumocystis cell wall beta-glucans stimulate alveolar epithelial cell chemokine generation through nuclear factor-kappaB-dependent mechanisms. AB - Exuberant inflammatory responses are associated with respiratory failure during Pneumocystis pneumonia. Alveolar epithelial cells (AECs) promote Pneumocystis attachment and proliferation, but also contribute prominently to host cytokine mediated inflammation during pneumonia. Recent investigations indicate that AECs produce macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-alpha (TNF-alpha) following challenge with Pneumocystis carinii. Nuclear factor-kappaB (NF-kappaB) is a ubiquitous transcription factor critical for regulation of proinflammatory cytokine expression. Herein, we assess rat AEC NF-kappaB responses to challenge with a P. carinii beta-glucan cell wall component (PCBG). Prominent nuclear translocation of p65 NF-kappaB was demonstrated following PCBG challenge. NF-kappaB activation was in part mediated through Protein Kinase C (PKC) signaling pathways. PCBG challenge of AECs was also shown to induce MIP-2 and TNF-alpha mRNA production, a response that was ameliorated by NF-kappaB inhibition. MIP-2 protein expression was also dramatically increased by PCBG challenge, in a manner that was significantly attenuated by both PKC and NF kappaB inhibition. The data further demonstrate that AEC chemokine responses were not mediated by the recently described dectin-1 receptor, but instead involved participation of cell surface lactosylceramide. These data support a significant role for AECs in host responses during Pneumocystis pneumonia, and further indicate that beta-glucan induces inflammatory cytokine production through NF kappaB-dependent mechanisms. PMID- 15746435 TI - Matrix metalloproteinase-9 modulation by resident arterial cells is responsible for injury-induced accelerated atherosclerotic plaque development in apolipoprotein E-deficient mice. AB - OBJECTIVE: Although matrix metalloproteinase-9 (MMP-9) has been implicated in atherosclerotic plaque instability, the exact role it plays in the plaque development and progression remains largely unknown. We generated apolipoprotein E (apoE)-deficient (apoE-/-) MMP-9-deficient (MMP-9-/-) mice to determine the mechanisms and the main cell source of MMP-9 responsible for the plaque composition during accelerated atherosclerotic plaque formation. METHODS AND RESULTS: Three weeks after temporary carotid artery ligation revealed that while on a Western-type diet, apoE-/- MMP-9-/- mice had a significant reduction in intimal plaque length and volume compared with apoE-/- MMP-9+/+ mice. The reduction in plaque volume correlated with a significantly lower number of intraplaque cells of resident cells and bone marrow-derived cells. To determine the cellular origin of MMP-9 in plaque development, bone marrow transplantation after total-body irradiation was performed with apoE-/- MMP-9+/+ and apoE-/- MMP 9-/- mice, which showed that only MMP-9 derived from resident arterial cells is required for plaque development. CONCLUSIONS: MMP-9 is derived from resident arterial cells and is required for early atherosclerotic plaque development and cellular accumulation in apoE-/- mice. PMID- 15746436 TI - Atherogenesis in mice does not require CD40 ligand from bone marrow-derived cells. AB - OBJECTIVE: Recent research suggests a central role for CD40 ligand (CD40L) in atherogenesis. However, the relevant cellular source of this proinflammatory cytokine remains unknown. To test the hypothesis that CD40L expressed on hematopoietic cell types (eg, macrophages, lymphocytes, platelets) is crucial to atherogenesis, we performed bone marrow reconstitution experiments using low density receptor-deficient (ldlr-/-) and ldlr-/-/cd40l-/- compound-mutant mice. METHODS AND RESULTS: As expected, systemic lack of CD40L in hypercholesterolemic ldlr-/- mice significantly reduced the development of atherosclerotic lesions in the aortic arch, aortic root, and abdominal aorta compared with ldlr-/- mice. Furthermore, atheromata in ldlr-/-/cd40l-/- mice showed reduced accumulation of macrophages and lipids and increased content in smooth muscle cells and collagen compared with ldlr-/- mice. Surprisingly, reconstitution of irradiated ldlr-/- mice with ldlr-/-/cd40l-/- bone marrow did not affect the size or composition of atherosclerotic lesions in the root or arch of hypercholesterolemic ldlr-/- mice. Moreover, lipid deposition in the abdominal aorta diminished only marginally compared with mouse aortas reconstituted with ldlr-/- bone marrow. CONCLUSIONS: These experiments demonstrate that CD40L modulates atherogenesis, at least in mice, primarily by its expression on nonhematopoietic cell types rather than monocytes, T lymphocytes, or platelets, a surprising finding with important pathophysiologic and therapeutic implications. PMID- 15746437 TI - Partial off-loading of longitudinal tension induces arterial tortuosity. AB - OBJECTIVES: Arterial tortuosity is a frequent manifestation of vascular disease and collateral vessel growth, but its causes are poorly understood. This study was designed to assess the relationship between the development of tortuosity and the mechanical forces that are imposed on arterial tissue. METHODS AND RESULTS: Axial strain in rabbit carotid arteries was reduced from 62+/-2% to 33+/-2% by implanting an interposition graft, prepared from the contralateral carotid, at the downstream end of the artery. Axial strain remained unchanged for 12 weeks; however, all vessels became tortuous because of tissue growth and remodeling. After 7 days, there was a marked elevation in proliferation rates of endothelial and smooth muscle cells; however, increased apoptosis was also detected, and no net accumulation of DNA was observed. Significant accumulations of elastin (24%) and total collagen (26%) occurred by 5 weeks. Gelatin zymography detected upregulation and activation of matrix metalloproteinase-2 (MMP-2), and confocal microscopy revealed enlargement of fenestrae in the internal elastic lamina. MMP inhibition by treatment with doxycycline prevented enlargement of fenestrae and development of tortuosity, and it enabled normalization of axial strain by 5 weeks. CONCLUSIONS: These findings indicate that substantial axial strain is necessary to sustain the morphological stability of arteries, and that a reduction in strain results in arterial tortuosity attributable to aberrant MMP activity. PMID- 15746438 TI - Association between serum uric acid, metabolic syndrome, and carotid atherosclerosis in Japanese individuals. AB - OBJECTIVE: There are few data available on possible independent association between uric acid and carotid atherosclerosis. Here we first sought to investigate association between uric acid levels and metabolic syndrome in Japanese; second, we assessed whether there is an independent association of uric acid with prevalence of carotid atherosclerosis in individuals subdivided according to gender and metabolic syndrome status. METHODS AND RESULTS: Cross sectional data from 8144 individuals who underwent general health screening were analyzed. After adjusting for age, total cholesterol, and smoking status, the odds ratios (95% CI) of sex-specific quartiles of serum uric acid for metabolic syndrome were 1.0, 1.06 (0.60 to 1.87), 2.18 (1.30 to 3.64), and 4.17 (2.56 to 6.79) in women, and 1.0, 0.92 (0.74 to 1.14), 1.52 (1.25 to 1.65), and 1.97 (1.61 to 2.40) in men. After adjusting for age, serum levels, total cholesterol, and smoking status, prevalence of carotid plaque was higher in subjects in the second, third, and fourth quartiles of uric acid level with odds ratios (95% CI) of 1.24 (1.01 to 1.52), 1.37 (1.11 to 1.68), and 1.31 (1.05 to 1.63), respectively, in men without metabolic syndrome but not in men with metabolic syndrome or in women with or without metabolic syndrome. CONCLUSIONS: The prevalence of metabolic syndrome showed a graded increase according to serum uric acid values in both genders. In men who did not have metabolic syndrome, uric acid was found to be an independent risk factor for incidence of carotid plaque. PMID- 15746440 TI - NO generation from nitrite and its role in vascular control. AB - NO generated from L-arginine by NO synthases (NOSs) in the endothelium and in other cells plays a central role in several aspects of vascular biology. The biological activity of NO is acutely terminated by oxidation to nitrite and nitrate, and these compounds have long been considered only as inert end-products of NO. However, this dogma is now being challenged because recent research convincingly has shown that the nitrite ion can be recycled back to bioactive NO again in blood and tissues. Nitrite reduction to NO can occur via several routes involving enzymes, proteins, vitamins, or even simple protons. This pathway may serve as a backup system for NO generation in conditions such as hypoxia, in which the NOS/L-arginine system is compromised, but detrimental effects can also be foreseen. With this new knowledge, nitrate and nitrite should probably be viewed as storage pools for NO rather than inert waste products. Here we discuss novel aspects of nitrite-dependent NO generation in vivo and its role in vascular control. PMID- 15746439 TI - Differential activation of mitogenic signaling pathways in aortic smooth muscle cells deficient in superoxide dismutase isoforms. AB - OBJECTIVE: Reactive oxygen species (ROS) integrate cellular signaling pathways involved in aortic smooth muscle cell (SMC) proliferation and migration associated with atherosclerosis. However, the effect of subcellular localization of ROS on SMC mitogenic signaling is not yet fully understood. METHODS AND RESULTS: We used superoxide dismutase (SOD)-deficient mouse aortic SMCs to address the role of subcellular ROS localization on SMC phenotype and mitogenic signaling. Compared with wild-type, a 54% decrease in total SOD activity (almost equal to 50% decrease in SOD1 protein levels) and a 42% reduction in SOD2 activity (approximately equal to 50% decrease in SOD2 protein levels) were observed in SOD1+/- and SOD2+/- SMCs, respectively. Consistent with this, basal and thrombin-induced superoxide levels increased in these SMCs. SOD1+/- and SOD2+/- SMCs exhibit increased basal proliferation and enhanced [3H]-thymidine and [3H]-leucine incorporation in basal and thrombin-stimulated conditions. Our results indicate preferential activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinases in SOD1+/- and janus kinase/signal transducer and activator of transcriptase (JAK/STAT) pathway in SOD2+/- SMCs. Pharmacological inhibitors of ERK1/2 p38 and JAK2 confirm the SOD genotype-dependent SMC proliferation. CONCLUSIONS: Our results suggest that SOD1 and SOD2 regulate SMC quiescence by suppressing divergent mitogenic signaling pathways, and dysregulation of these enzymes under pathophysiological conditions may lead to SMC hyperplasia and hypertrophy. PMID- 15746441 TI - Impaired KCNQ1-KCNE1 and phosphatidylinositol-4,5-bisphosphate interaction underlies the long QT syndrome. AB - Nearly a hundred different KCNQ1 mutations have been reported as leading to the cardiac long QT syndrome, characterized by prolonged QT interval, syncopes, and sudden death. We have previously shown that phosphatidylinositol-4,5-bisphosphate (PIP2) regulates the KCNQ1-KCNE1 complex. In the present study, we show that PIP2 affinity is reduced in three KCNQ1 mutant channels (R243H, R539W, and R555C) associated with the long QT syndrome. In giant excised patches, direct application of PIP2 on the cytoplasmic face of the three mutant channels counterbalances the loss of function. Reintroduction of a positive charge by application of methanethiosulfonate ethylammonium on the cytoplasmic face of R555C mutant channels also restores channel activity. The channel affinity for a soluble analog of PIP2 is decreased in the three mutant channels. By using a model that describes the KCNQ1-KCNE1 channel behavior and by fitting the relationship between the kinetics of deactivation and the current amplitude obtained in whole-cell experiments, we estimated the PIP2 binding and dissociation rates on wild-type and mutant channels. The dissociation rate of the three mutants was higher than for the wild-type channel, suggesting a decreased affinity for PIP2. PIP2 binding was magnesium-dependent, and the PIP2-dependent equilibrium constant in the absence of magnesium was higher with the wild-type than with the mutant channels. Altogether, our data suggest that a reduced PIP2 affinity of KCNQ1 mutants can lead to the long QT syndrome. PMID- 15746442 TI - NAD(P)H oxidase inhibition attenuates neuronal chronotropic actions of angiotensin II. AB - It is well established that the central cardiovascular effects of angiotensin II (Ang II) involve superoxide production. However, the intracellular mechanism by which reactive oxygen species (ROS) signaling regulates neuronal Ang II actions remains to be elucidated. In the present study, we have used neuronal cells in primary cultures from the hypothalamus and brain stem areas to study the role of ROS on the cellular actions of Ang II. Ang II increases neuronal firing rate, an effect mediated by the AT(1) receptor subtype and involving inhibition of the delayed rectifier potassium current (I(Kv)). This increase in neuronal activity was associated with increases in NADPH oxidase activity and ROS levels within neurons, the latter evidenced by an increase in ethidium fluorescence. The increases in NADPH oxidase activity and ethidium fluorescence were blocked by either the AT(1) receptor antagonist losartan or by the selective NAD(P)H oxidase inhibitor gp91ds-tat. Extracellular application of the ROS scavenger, Tempol, attenuated the Ang II-induced increase in neuronal firing rate by 70%. In addition, gp91ds-tat treatment resulted in a 50% inhibition of Ang II-induced increase in firing rate. In contrast, the ROS generator Xanthine-Xanthine oxidase significantly increased neuronal firing rate. Finally, Ang II inhibited neuronal I(Kv,) and this inhibition was abolished by gp91ds-tat treatment. These observations demonstrate, for the first time, that Ang II regulates neuronal activity via a series of events that includes ROS generation and inhibition of I(Kv). This signaling seems to be a critical cellular event in central Ang II regulation of cardiovascular function. PMID- 15746443 TI - Enhancement of cardiac function and suppression of heart failure progression by inhibition of protein phosphatase 1. AB - Abnormal calcium cycling, characteristic of experimental and human heart failure, is associated with impaired sarcoplasmic reticulum calcium uptake activity. This reflects decreases in the cAMP-pathway signaling and increases in type 1 phosphatase activity. The increased protein phosphatase 1 activity is partially due to dephosphorylation and inactivation of its inhibitor-1, promoting dephosphorylation of phospholamban and inhibition of the sarcoplasmic reticulum calcium-pump. Indeed, cardiac-specific expression of a constitutively active inhibitor-1 results in selective enhancement of phospholamban phosphorylation and augmented cardiac contractility at the cellular and intact animal levels. Furthermore, the beta-adrenergic response is enhanced in the transgenic hearts compared with wild types. On aortic constriction, the hypercontractile cardiac function is maintained, hypertrophy is attenuated and there is no decompensation in the transgenics compared with wild-type controls. Notably, acute adenoviral gene delivery of the active inhibitor-1, completely restores function and partially reverses remodeling, including normalization of the hyperactivated p38, in the setting of pre-existing heart failure. Thus, the inhibitor 1 of the type 1 phosphatase may represent an attractive new therapeutic target. PMID- 15746444 TI - Common variants in myocardial ion channel genes modify the QT interval in the general population: results from the KORA study. AB - Altered myocardial repolarization is one of the important substrates of ventricular tachycardia and fibrillation. The influence of rare gene variants on repolarization is evident in familial long QT syndrome. To investigate the influence of common gene variants on the QT interval we performed a linkage disequilibrium based SNP association study of four candidate genes. Using a two step design we analyzed 174 SNPs from the KCNQ1, KCNH2, KCNE1, and KCNE2 genes in 689 individuals from the population-based KORA study and 14 SNPs with results suggestive of association in a confirmatory sample of 3277 individuals from the same survey. We detected association to a gene variant in intron 1 of the KCNQ1 gene (rs757092, +1.7 ms/allele, P=0.0002) and observed weaker association to a variant upstream of the KCNE1 gene (rs727957, +1.2 ms/allele, P=0.0051). In addition we detected association to two SNPs in the KCNH2 gene, the previously described K897T variant (rs1805123, -1.9 ms/allele, P=0.0006) and a gene variant that tags a different haplotype in the same block (rs3815459, +1.7 ms/allele, P=0.0004). The analysis of additive effects by an allelic score explained a 10.5 ms difference in corrected QT interval length between extreme score groups and 0.951 of trait variance (P<0.00005). These results confirm previous heritability studies indicating that repolarization is a complex trait with a significant heritable component and demonstrate that high-resolution SNP-mapping in large population samples can detect and fine map quantitative trait loci even if locus specific heritabilities are small. PMID- 15746445 TI - Endogenous p53 protects vascular smooth muscle cells from apoptosis and reduces atherosclerosis in ApoE knockout mice. AB - Recent studies have indicated that the tumor suppressor gene p53 limits atherosclerosis in animal models; p53 expression is also increased in advanced human plaques compared with normal vessels, where it may induce growth arrest and apoptosis. However, controversy exists as to the role of endogenous levels of p53 in different cell types that comprise plaques. We examined atherosclerotic plaque development and composition in brachiocephalic arteries and aortas of p53-/-/ApoE /- mice versus wild type p53 controls. p53-/- mice demonstrated increased aortic plaque formation, with increased rates of cell proliferation and reduced rates of apoptosis in brachiocephalic arteries. Although most proliferating cells were monocyte/macrophages, apoptotic cells were both vascular smooth muscle cells (VSMCs) and macrophages. Transplant of p53 bone marrow to p53-/-/ApoE-/- mice reduced aortic plaque formation and cell proliferation in brachiocephalic plaques, but also markedly reduced apoptosis. To examine p53 regulation of these processes, we studied proliferation and apoptosis in macrophages, bone marrow stromal cells and VSMCs cultured from these mice. Although endogenous p53 promoted apoptosis in macrophages, it protected VSMCs and stromal cells from death, a hitherto unknown function in these cells, in part by inhibiting DNA damage response enzymes. p53 also inhibited stromal cell expression of VSMC markers. We conclude that endogenous levels of p53 protect VSMCs and stromal cells against apoptosis, while promoting apoptosis in macrophages, and protect against atherosclerosis development. PMID- 15746446 TI - An intimate relationship: Ca2+ and cardiac ion channels. PMID- 15746447 TI - Regulating protective immunity in atherosclerosis. PMID- 15746448 TI - Multi-tasking RGS proteins in the heart: the next therapeutic target? AB - Regulator of G-protein-signaling (RGS) proteins play a key role in the regulation of G-protein-coupled receptor (GPCR) signaling. The characteristic hallmark of RGS proteins is a conserved approximately 120-aa RGS region that confers on these proteins the ability to serve as GTPase-activating proteins (GAPs) for G(alpha) proteins. Most RGS proteins can serve as GAPs for multiple isoforms of G(alpha) and therefore have the potential to influence many cellular signaling pathways. However, RGS proteins can be highly regulated and can demonstrate extreme specificity for a particular signaling pathway. RGS proteins can be regulated by altering their GAP activity or subcellular localization; such regulation is achieved by phosphorylation, palmitoylation, and interaction with protein and lipid-binding partners. Many RGS proteins have GAP-independent functions that influence GPCR and non-GPCR-mediated signaling, such as effector regulation or action as an effector. Hence, RGS proteins should be considered multifunctional signaling regulators. GPCR-mediated signaling is critical for normal function in the cardiovascular system and is currently the primary target for the pharmacological treatment of disease. Alterations in RGS protein levels, in particular RGS2 and RGS4, produce cardiovascular phenotypes. Thus, because of the importance of GPCR-signaling pathways and the profound influence of RGS proteins on these pathways, RGS proteins are regulators of cardiovascular physiology and potentially novel drug targets as well. PMID- 15746449 TI - Early recanalization rates and clinical outcomes in patients with tandem internal carotid artery/middle cerebral artery occlusion and isolated middle cerebral artery occlusion. AB - BACKGROUND: Patients with isolated middle cerebral artery (MCA) and tandem MCA/internal carotid artery (ICA) obstruction have similar presenting symptoms and stroke severity. We aimed to investigate early recanalization of MCA and clinical outcomes in patients with tandem ICA/MCA obstructions and isolated MCA occlusion. METHODS: Patients with MCA occlusion on pretreatment transcranial Doppler were treated with intravenous tissue plasminogen activator. ICA lesions were detected with carotid duplex. Early neurological improvement (ENI) was defined by reduction of National Institutes of Health Stroke Scale (NIHSS) > or =10 points or total NIHSS < or =3 points. Good outcome at 3 months was a modified Rankin score of < or =2. RESULTS: Among 104 patients, 31% had tandem lesions and 69% had isolated MCA occlusions. Complete recanalization rate was 39% in isolated MCA occlusion group and 9% in tandem group (P=0.002). ENI at 24 hours occurred in 46% of the isolated MCA occlusion group and in 25% of the tandem group (P=0.045). Good outcome was achieved by 30% and 39% respective patients (NS). CONCLUSIONS: The tandem lesion group showed lower early recanalization rate and ENI rate than the isolated MCA occlusion group. Despite this, good outcomes were similar in both groups. PMID- 15746450 TI - Blood pressure control and risk of stroke: a population-based prospective cohort study. AB - BACKGROUND AND PURPOSE: Adequate control of blood pressure (BP) is a cornerstone in stroke prevention. This study explored the risk of stroke in relation to the quality of BP control in a population-based cohort and whether control of hypertension was related to background characteristics of patients. METHODS: A total of 27,936 subjects (10,953 men and 16,983 women), 45 to 73 years old, living in Malmo, Sweden participated in the study. Incidence of stroke was followed-up for a mean period of 6 years. Controlled BP was defined as BP <140/90 mm Hg in subjects with pharmacological treatment for hypertension. RESULTS: In the whole cohort, 16 648 subjects (60%) had hypertension (BP > or =140/90 mm Hg) and 23% of them received treatment. Among treated hypertensives, 88.2% had BP levels > or =140/90 mm Hg and 49.5% had BP levels >or =160/100 mm Hg. During the follow-up, 137 strokes occurred among treated hypertensive subjects. The crude incidence of stroke was 289/100 000 person-year in controlled hypertensive subjects and 705/100,000 person-year in treated hypertensive subjects with BP >or =140/90 mm Hg. It was estimated that approximately 45% of all strokes among subjects with treatment for hypertension might be attributed to uncontrolled BP. In treated hypertensives, the risk of stroke increased significantly with advancing age, current smoking, high level of diastolic BP, and diabetes. In hypertensive subjects without treatment (n=12 819), incidence of stroke was 363/100,000 person-year. CONCLUSIONS: Uncontrolled BP is highly prevalent in patients with pharmacological treatment for hypertension. More than 90% of stroke in this group occurred in those with uncontrolled BP. Adequate hypertension control may prevent a substantial proportion of first-ever stroke among treated hypertensives. PMID- 15746451 TI - Common carotid intima-media thickness and risk of acute myocardial infarction: the role of lumen diameter. AB - BACKGROUND: It has been argued that lumen diameter of the common carotid artery should be taken into account in analyses on common carotid intima-media thickness (CIMT) and cardiovascular risk. Yet, no published report has dealt with this issue in detail. METHODS: In the Rotterdam study baseline ultrasound images of the carotid arteries were made. During follow-up of 8.2 years, 656 new acute myocardial infarctions (AMI) occurred. Regression analysis was used to study myocardial infarction relation to right (or left) common CIMT with various adjustments for right-, or left-sided lumen diameter. Lumen adjustment was made by (1) a simple adjustment in a regression equation; (2) using the CIMT-to-lumen ratio; (3) using arterial mass, calculated as ({pix[(lumen+near wall CIMT+far wall CIMT)/2]2}-[pix(lumen/2)2]). RESULTS: AMI disease risk increased per standard deviation increase in common CIMT (0.177 mm): hazard ratio (HR) 1.28 (95% CI, 1.19 to 1.37). When lumen diameter was taken into account the HR was 1.26 (95% CI, 1.18 to 1.35). The HR for the CIMT-to-lumen ratio was 1.18 (95% CI, 1.11 to 1.27) and for arterial mass 1.28 (95% CI, 1.19 to 1.37). Additional analyses indicated that the CIMT-to-lumen ratio at lower CIMT levels appears to reflect arterial remodelling rather than risk of cardiovascular disease. CONCLUSIONS: We conclude that using the CIMT-to-lumen ratio yields the weakest associations. Other approaches for adjustment for common carotid lumen diameter do not affect the magnitude or precision of the association of common CIMT to risk of AMI. When the interest is in risk relations the preference goes to either CIMT or arterial mass measurement. PMID- 15746452 TI - Functional recovery in aged and young rats after embolic stroke: treatment with a phosphodiesterase type 5 inhibitor. AB - BACKGROUND AND PURPOSE: Advanced age is associated with a decrease in brain plasticity compared with the young adult. Sildenafil, a phosphodiesterase type 5 (PDE5) inhibitor promotes brain plasticity and improves functional outcome after stroke in the young animal. Here, we test the hypothesis that sildenafil provides restorative therapeutic benefit to the aged animal. METHODS: Male Wistar rats (aged, 18-month old; young, 3-month old) were subjected to embolic stroke. Saline or sildenafil was administered daily at a dose of 2 mg/kg orally or 10 mg/kg subcutaneously for 7 consecutive days starting 24 hour after stroke onset. RESULTS: Aged rats exhibited significant impairment of functional recovery and reductions of vascular density, and endothelial cell proliferation compared with young rats. Aged rats treated with sildenafil at a dose of 10 mg/kg but not 2 mg/kg, showed significant improvements of functional recovery and concomitant increases in cortical cyclic guanosine 3',5'-cyclic monophosphate (cGMP) level, vascular density, endothelial cell proliferation, and synaptogenesis compared with aged rats treated with saline. In young rats, treatment with sildenafil at a dose of 2 or 10 mg/kg significantly enhanced functional recovery and amplified brain plasticity compared with young rats treated with saline. CONCLUSIONS: Age is associated with reduction of angiogenesis, and poor neurological functional recovery after stroke. However, treatment of aged stroke rats with sildenafil improves functional recovery that is likely fostered by enhancement of angiogenesis and synaptogenesis. PMID- 15746454 TI - Safety and efficacy of endovascular treatment of carotid artery stenosis compared with carotid endarterectomy: a Cochrane systematic review of the randomized evidence. AB - BACKGROUND AND PURPOSE: Endovascular treatment of atherosclerotic carotid artery stenosis may be an alternative to surgical endarterectomy. To evaluate the safety and efficacy of endovascular techniques, we conducted a systematic review of randomized studies that compared endovascular treatment with surgery for carotid stenosis. METHODS: We searched the Cochrane Stroke Group trials register, the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, and Science Citation Index for randomized trials of carotid angioplasty and/or stenting compared with surgery. We also contacted researchers in the field and balloon catheter and stent manufacturers. RESULTS: Five trials involving 1269 patients were included. Analysis of 30-day safety data found no significant difference in the odds of treatment-related death or any stroke (odds ratio [OR], endovascular surgery, 1.33; 95% confidence interval [CI], 0.86 to 2.04), death or disabling stroke (OR, 1.22; CI, 0.61 to 2.41), or death, any stroke, or myocardial infarction (OR, 1.04; CI, 0.69 to 1.57). At 1 year after randomization, there was no significant difference between the 2 treatments in the rate of any stroke or death (OR, 1.01; CI, 0.71 to 1.44). Endovascular treatment significantly reduced the risk of cranial nerve injury (OR, 0.13; CI, 0.06 to 0.25). There was substantial heterogeneity between the trials for 4 of the 5 outcomes. CONCLUSIONS: No significant difference in the major risks of treatment was found but the wide confidence intervals indicate that it is not possible to exclude a difference in favor of one treatment. Minor complication rates favor endovascular treatment. There is currently insufficient evidence to support a widespread change in clinical practice away from recommending carotid endarterectomy as the treatment of choice for suitable carotid artery stenosis. Patients suitable for carotid endarterectomy should only be offered stenting within the ongoing randomized trials of stenting versus surgery. PMID- 15746455 TI - Family history of stroke in patients with transient ischemic attack in relation to hypertension and other intermediate phenotypes. AB - BACKGROUND AND PURPOSE: Family history of stroke (FHx(stroke)) is a risk factor for ischemic stroke, but there are insufficient data on the relationship with stroke subtypes and intermediate phenotypes (IPs), such as hypertension. Specifically, there are no reliable data on the associations of FHx(stroke) in patients with transient ischemic attack (TIA) in whom relationships with IPs are likely to be determined most reliably. METHODS: We studied FHx(stroke) and FHx of myocardial infarction (FHx(MI)) in TIA patients from 2 population-based incidence studies and 2 prospective consecutive hospital-referred series. We related the presence of FHx to baseline characteristics, clinical subtype, and IPs. RESULTS: Results were similar in the 4 cohorts, and so data on all 783 patients were pooled. FHx(stroke) was less common than FHx(MI) (189 versus 254; P=0.0003). FHx(stroke) and FHx(MI) were strongly related to history of hypertension in the proband (odds ratio [OR], 1.78; 95% CI, 1.28 to 2.48; P=0.0008; and OR, 2.10, 95% CI, 1.55 to 2.85; P<0.0001, respectively). Highest recorded premorbid systolic and diastolic blood pressures (mm Hg) were significantly higher in cases with FHx(stroke) than those without and increased with the number of affected first degree relatives (0 181/100; 1 185/104; > or =2 198/109; P=0.03). There was no association between FHx(stroke) and age, diabetes, smoking, plasma glucose, cholesterol, or territory of TIA, but FHx(stroke) was less common in patients with ocular TIA than in cases with cerebral TIA (OR, 0.53; 95% CI, 0.34 to 0.82; P=0.004), although the association was no longer significant after adjustment for hypertension. CONCLUSIONS: The strong association between hypertension and FHx(stroke) suggests that familial susceptibility to cerebral ischemia is attributable, at least partly, to familial predisposition to hypertension. This should be taken into account in studies of the genetics of ischemic stroke. PMID- 15746456 TI - Relations of serum high-sensitivity C-reactive protein and interleukin-6 levels with silent brain infarction. AB - BACKGROUND AND PURPOSE: Small silent brain infarction (SBI) is often found on magnetic resonance (MR) images of apparently healthy individuals at cardiovascular risk. Particularly, small SBI found in subcortical white matter, basal ganglia, or thalamus is thought to be caused by cerebral small vessel disease. Although several lines of evidence suggest a role of inflammatory processes in atherothrombotic vascular events, their involvement in SBI remains to be determined. This study examines the associations between serum inflammatory markers and SBI as a manifestation of cerebral small vessel disease. METHODS: One hundred ninety-four patients without histories of cardiovascular accidents were prospectively enrolled for this study. All patients underwent brain MR imaging and carotid ultrasonography, and patients with SBI diagnosed underwent further MR angiography. As common inflammatory markers, serum levels of high-sensitivity C reactive protein (hsCRP) and interleukin-6 (IL-6) were evaluated. RESULTS: SBIs were found in 40 patients, and all of those were located in subcortical and infratentorial area, without MR angiographic evidence for obstructive lesions in proximal cerebral arteries. Mean hsCRP and IL-6 levels were higher in patients with SBI than in those without. Also, higher levels of both hsCRP (odds ratio [OR], 1.85 per standard deviation [SD] increase) and IL-6 (OR, 2.00/SD increase) were associated with higher likelihood for SBI. Moreover, the associations were only slightly attenuated when adjusting traditional cardiovascular risk factors and carotid IMT. CONCLUSIONS: Higher levels of hsCRP and IL-6 appear to be associated with small SBI, suggesting a role of inflammatory processes in cerebral small vessel disease. PMID- 15746457 TI - Markedly reduced apparent blood volume on bolus contrast magnetic resonance imaging as a predictor of hemorrhage after thrombolytic therapy for acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Accurate assessment of the risk of hemorrhage could help to improve patient selection for thrombolytic therapy and reduce hemorrhagic complications, especially for patients with longer or uncertain time after symptom onset. This study sought to define characteristics of hemodynamic magnetic resonance imaging (MRI), which best predict hemorrhage. METHODS: Bolus contrast and diffusion MRI were performed before intravenous tissue plasminogen activator (tPA) therapy in 20 patients presenting with acute stroke symptoms within the first 6 hours after symptom onset. Hemorrhage was assessed on follow up MRI (n=15) and computed tomography (n=5) scans. RESULTS: Of the 20 patients studied, 5 had detectable hemorrhage on follow-up scans. Blood volume maps demonstrated virtually no signal within much of the hemorrhagic region, indicating contrast did not arrive by the end of the imaging series (80 seconds). Within the hemodynamically abnormal region, a threshold of at least 126 voxels with blood volume <5% of contralateral normal gray matter separated hemorrhagic patients from nonhemorrhagic with a sensitivity of 100% and a specificity of 73% (P<0.01). All subjects with hemorrhage were at least partially reperfused after thrombolysis, whereas most false-positives did not reperfuse (P<0.05). The number of low blood volume voxels within individual patients correlated with the number of voxels with apparent diffusion coefficient values <550x10(-6) mm2/s (P<0.019), another previously proposed predictor of hemorrhage. CONCLUSIONS: Extremely low or completely absent contrast arrival may indicate tissue-at-risk for hemorrhage before tPA treatment and thus may aid in risk-benefit assessments. Occurrence of hemorrhage within at-risk areas may depend on tissue reperfusion. PMID- 15746458 TI - Endovascular stent-assisted angioplasty in the management of traumatic internal carotid artery dissections. AB - BACKGROUND AND PURPOSE: The prognosis of traumatic dissection of the internal carotid artery is worse than for spontaneous dissections. Rapid stenting followed by antiplatelet therapy may prevent complications when anticoagulation therapy is not applicable. METHODS: Patients with angiographically proven traumatic carotid artery dissection and hemodynamic significant hemispheric hypoperfusion, or in whom anticoagulant therapy was either contraindicated or failed clinically, were regarded as being at high risk for stroke and were selected for stenting. RESULTS: Ten patients with traumatic dissection underwent stenting. Endovascular treatment reduced mean dissection stenosis from 69% to 8%. During a mean clinical follow-up time of 16 months, none had additional transient ischemic attacks or stroke. Doppler ultrasound studies did not detect any signs of de novo in-stent stenosis. CONCLUSIONS: In selected cases of traumatic carotid artery dissections, endovascular stent-assisted angioplasty immediately restored the integrity of the vessel lumen and prevented efficiently the occurrence of new ischemic events, without additional anticoagulation. PMID- 15746459 TI - Monocyte count is a predictor of novel plaque formation: a 7-year follow-up study of 2610 persons without carotid plaque at baseline the Tromso Study. AB - BACKGROUND AND PURPOSE: Activation of monocytes and differentiation into lipid laden macrophages are fundamental events in generation of atherosclerotic lesions. There exist few data on monocyte activity and the risk for atherosclerosis. In this prospective population-based study, we examined whether monocyte count in blood is a predictor of future plaque formation in persons without pre-existing carotid atherosclerosis. METHODS: At baseline, we measured monocyte count, white cell count (WCC), fibrinogen, intima-media thickness (IMT), and traditional cardiovascular risk factors in 2610 men and women aged 25 to 82 years who on ultrasound had no plaque in their right carotid artery. After 7 years of follow-up, a new ultrasound screening was performed and the number of novel plaques was grouped as none, 1 plaque, and 2 or more plaques. RESULTS: In multivariate analysis, monocyte count, age, sex, total cholesterol, current smoking, systolic blood pressure, and IMT were independent predictors of novel plaque formation. No significant association was found between plaque formation and either WCC or fibrinogen. For 1 standard deviation (0.17x10(9)) increase in monocyte count, the risk of being in a higher plaque category increased by 18% (OR, 1.18; 95% CI, 1.08 to 1.29). In the highest monocyte quartile, the risk for having plaque compared with the lowest quartile was 1.85 (OR) (95% confidence interval, 1.41 to 2.43). Repeating the analysis without IMT did not change the monocyte estimate. Excluding subjects with cardiovascular disease and diabetes mellitus from analysis neither changed the monocyte estimate. CONCLUSIONS: Monocyte count is an independent predictor of future plaque formation in subjects without pre-existing carotid atherosclerosis. PMID- 15746460 TI - Identification of embolic stroke patterns by diffusion-weighted MRI in clinically defined lacunar stroke syndromes. AB - BACKGROUND: A number of clinical syndromes describing the presentation of deep brain infarcts are called lacunar syndromes resulting from small vessel occlusion (SVO). To verify the reliability of the clinical diagnosis "lacunar syndrome," the value was investigated with diffusion-weighted MRI (DWI). METHODS AND RESULTS: A total of 73 patients (mean age 66 years; range 35 to 83 years) with sudden onset of a classical lacunar syndrome were enrolled. On the basis of the DWI findings, patients were divided into 3 groups: group 1, single subcortical lesion (<15-mm lesion; 43 patients; 59%); group 2, large (> or =15 mm) or scattered lesions in 1 vascular territory (16 patients; 22%); and group 3, multiple lesions in multiple vascular territories (14 patients; 19%). A stroke mechanism other than SVO could be identified in 17 (23%) patients. Cardiac work up revealed a cardiac embolic source in 8 patients (11%). Duplex sonography revealed symptomatic stenosis in 9 patients (12%). Based on the work-up information, 29 patients (40%) were found to have a potential cause of stroke other than SVO. A significant correlation with >1 single lesion on DWI-MRI and a clinical proven embolic source was observed (P=0.002). In 9 patients with MRI suspicious for a pathomechanism other than SVO, no embolic source was found. CONCLUSIONS: The use of DWI-MRI improves the accuracy of the subtype diagnosis of stroke. Inaccuracy has to be expected in approximately one third if lacunar diagnosis is based on clinical and computed tomography findings. Most of these "false-positive" cases are attributable to large artery or cardiogenic embolic stroke. PMID- 15746461 TI - Psychosocial impact of screening for intracranial aneurysms in relatives with familial subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: In families with > or =2 relatives with intracranial aneurysms (IAs), screening for IAs in asymptomatic first-degree relatives is often recommended. We assessed the long-term psychosocial impact of such screening. METHODS: We identified all persons with IA (screen-positives) and matched them for age and sex with 2 controls without IA (screen-negatives) from hospital-based registers of familial IA. Persons underwent telephone interviews using questionnaires that covered the areas of psychosocial impact of screening, health-related quality of life (HRQoL), and mood. Data were compared between screen-positives and screen-negatives, and with reference populations. RESULTS: Overall, 105 persons from 33 families with IA were included, of whom 35 were screen-positive and 70 were screen-negative. Of the screen-positives, 12 (44%) had reduced their work and 23 (66%) had experienced changes in > or =1 area of independence, self-esteem, future outlook, or personal relationships. In contrast, only 1 (2%) screen-negative person had stopped working and 12 (17%) others had experienced changes in their self-esteem, future outlook, or relationships. Screen-positives had lower HRQoL compared with screen-negatives and the reference population, whereas both screen groups had higher mean depression scores than the reference population. Despite these effects, only 3 persons regretted participating in screening. CONCLUSIONS: Although screening for IA is an important preventative strategy in high-risk individuals, it is associated with considerable psychosocial effects, both positive and negative. Greater awareness of such outcomes, and appropriate intervention where necessary, would appear to be a necessary component of IA screening programs. PMID- 15746462 TI - Adenovirus-mediated gene transfer of heparin-binding epidermal growth factor-like growth factor enhances neurogenesis and angiogenesis after focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: Recent studies have demonstrated that neurotrophic factors promote neurogenesis after cerebral ischemia. However, it remains unknown whether administration of genes encoding those factors could promote neural regeneration in the striatum and functional recovery. Here, we examined the efficacy of intraventricular injection of a recombinant adenovirus-expressing heparin-binding epidermal growth factor-like growth factor (HB-EGF) on neurogenesis, angiogenesis, and functional outcome after focal cerebral ischemia. METHODS: Transient focal ischemia was induced by middle cerebral artery occlusion (MCAO) for 80 minutes with a nylon filament in Wistar rats. Three days after MCAO, either adenovirus-expressing HB-EGF (Ad-HB-EGF) or Ad-LacZ, the control vector, was injected into the lateral ventricle on the ischemic side. Bromodeoxyuridine (BrdU) was injected intraperitoneally twice daily on the sixth and seventh days. On the eighth or 28th day after MCAO, we evaluated infarct volume, neurogenesis, and angiogenesis histologically. Neurological outcome was serially evaluated by the rotarod test after MCAO. RESULTS: There was no significant difference in infarct volume between the 2 groups. Treatment with Ad HB-EGF significantly increased the number of BrdU-positive cells in the subventricular zone on the 8th day. In addition, on the 28th day, BrdU-positive cells differentiated into mature neurons in the striatum on the ischemic side but seldom the cells given Ad-LacZ. Enhancement of angiogenesis at the peri-infarct striatum was also observed on the eighth day in Ad-HB-EGF-treated rats. Treatment with Ad-HB-EGF significantly enhanced functional recovery after MCAO. CONCLUSIONS: Our data suggest that gene therapy using Ad-HB-EGF contributes to functional recovery after ischemic stroke by promoting neurogenesis and angiogenesis. PMID- 15746463 TI - Cilostazol prevents the progression of the symptomatic intracranial arterial stenosis: the multicenter double-blind placebo-controlled trial of cilostazol in symptomatic intracranial arterial stenosis. AB - BACKGROUND AND PURPOSE: Cilostazol, a phosphodiesterase inhibitor, has been reported to reduce restenosis rate after coronary angioplasty and stenting. This study was performed to investigate the effect of cilostazol on the progression of intracranial arterial stenosis (IAS). METHODS: We randomized 135 patients with acute symptomatic stenosis in the M1 segment of middle cerebral artery or the basilar artery to either cilostazol 200 mg per day or placebo for 6 months. Aspirin 100 mg per day was also given to all patients. Patients with potential embolic sources in the heart or extracranial arteries were excluded. IAS was assessed by magnetic resonance angiogram (MRA) and transcranial Doppler (TCD) at the time of recruitment and 6 months later. The primary outcome was the progression of symptomatic IAS on MRA and secondary outcomes were clinical events and progression on TCD. RESULTS: Thirty-eight patients were prematurely terminated. Dropout rates and reasons for dropouts were similar between the cilostazol and placebo groups. There was no stroke recurrence in either cilostazol or placebo group, but there was 1 death and 2 coronary events in each group. In cilostazol group, 3 (6.7%) of 45 symptomatic IAS progressed and 11 (24.4%) regressed. In placebo group, 15 (28.8%) of symptomatic IAS progressed and 8 (15.4%) regressed. Progression of symptomatic IAS in cilostazol group was significantly lower than that in placebo group (P=0.008) CONCLUSIONS: Our study suggests that symptomatic IAS is a dynamic lesion and cilostazol may prevent its progression. PMID- 15746464 TI - Adrenomedullin enhances therapeutic potency of mesenchymal stem cells after experimental stroke in rats. AB - BACKGROUND AND PURPOSE: Adrenomedullin (AM) induces angiogenesis and inhibits cell apoptosis through the phosphatidylinositol 3-kinase/Akt pathway. Transplantation of mesenchymal stem cells (MSCs) has been shown to improve neurological deficits after stroke in rats. We investigated whether AM enhances the therapeutic potency of MSC transplantation. METHODS: Male Lewis rats (n=100) were subjected to 2-hour middle cerebral artery occlusion. Immediately after reperfusion, rats were assigned randomly to receive intravenous transplantation of MSCs plus subcutaneous infusion of AM for 7 days (MSC+AM group), AM infusion alone (AM group), MSC transplantation alone (MSC group), or vehicle infusion (control group). Neurological and immunohistological assessments were performed to examine the effects of these treatments. RESULTS: Some engrafted MSCs were positive for neuronal and endothelial cell markers, although the number of differentiated MSCs did not differ significantly between the MSC and MSC+AM groups. The neurological score significantly improved in the MSC, AM, and MSC+AM groups compared with the control group. Importantly, improvement in the MSC+AM group was significantly greater than that in the MSC and AM groups. There was marked induction of angiogenesis in the ischemic penumbra in the MSC+AM group, followed by the AM, MSC, and control groups. AM infusion significantly inhibited apoptosis of transplanted MSCs. As a result, the number of engrafted MSCs in the MSC+AM group was significantly higher than that in the MSC group. CONCLUSIONS: AM enhanced the therapeutic potency of MSCs, including neurological improvement, possibly through inhibition of MSC apoptosis and induction of angiogenesis. PMID- 15746465 TI - Study of the relationship between social deprivation and outcome after stroke. AB - BACKGROUND AND PURPOSE: Although the incidence and mortality of stroke are known to be inversely related to socioeconomic status (SES), the relationship between SES and recovery after stroke has been little-studied. This study has investigated the relationship between SES and case fatality, "death or dependency," and "death or institutional care" at 6 months after stroke. METHODS: Patients with acute stroke (n=2709) were identified using routine hospital discharge data and SES was measured using Carstairs scores (an ecological index of social deprivation). Case mix and treatment data were collected by medical chart review, case fatality by record linkage, and functional status and place of residence by questionnaire. Logistic regression was used to adjust the association of social deprivation and outcome for case mix and selected treatment variables. RESULTS: With increasing social deprivation, patients were younger, more likely to live alone, and, on admission, more likely to need help to walk. Social deprivation was not associated with case fatality or with "death or institutional care" in any analysis. However, patients residing in the most deprived areas (deprivation categories 6 and 7) were significantly more likely to be dead or dependent than patients from more affluent areas. This association was weakened but remained after adjusting for case mix and treatment variables. CONCLUSIONS: These findings contribute to growing evidence of an inverse social gradient in disability after stroke. Institutionalization, as a proxy for functional outcome, may not reflect this fact. A marked social gradient in case fatality after stroke seems unlikely. PMID- 15746466 TI - Pregnancy outcomes following hospitalization for motor vehicle crashes in Washington State from 1989 to 2001. AB - This retrospective cohort study evaluated the risk of adverse pregnancy outcomes following motor vehicle crashes during pregnancy. The authors assessed outcomes of pregnant women hospitalized for motor vehicle crashes in Washington State from 1989 to 2001 (n = 582). They used the Injury Severity Score (ISS) to classify 84 severely injured (ISS > or =9), 309 non-severely injured (ISS 1-8), and 189 uninjured (ISS 0) pregnant women and compared them with pregnant women who had not been hospitalized for a motor vehicle crash (n = 17,274). Of pregnant women in motor vehicle crashes, 82.9% were hospitalized and discharged without delivering, and 17.1% delivered at hospitalization. Compared with women not in motor vehicle crashes, severely and non-severely injured women were at increased risk of placental abruption and cesarean delivery, and their infants were at increased risk of respiratory distress syndrome and fetal death. Uninjured women were also at increased risk of preterm labor (relative risk = 7.9, 95% confidence interval: 6.4, 9.8) and placental abruption (relative risk = 6.6, 95% confidence interval: 3.9, 11.2) compared with women not in motor vehicle crashes. Pregnant women hospitalized following motor vehicle crashes are at increased risk of adverse pregnancy outcomes, regardless of the presence or severity of injuries. PMID- 15746467 TI - Use of discretionary protective equipment and rate of lower extremity injury in high school athletes. AB - Use of protective equipment is an important sports injury prevention strategy, yet use of protective equipment by high school athletes has seldom been studied. The authors analyzed data from a 3-year (1996-1999), stratified, two-stage cluster sample of athletes from 12 organized sports in 100 North Carolina high schools (n = 19,728 athlete-seasons). Information on each athlete's use of protective equipment and prior injury was collected during the preseason. Prospective information on injuries and weekly participation in games and practices was collected during the playing season. Use of lower extremity discretionary protective equipment tended to decrease the overall rate of lower extremity injury (rate ratio (RR) = 0.91, 95% confidence interval (CI): 0.72, 1.15). However, this slight protective effect was entirely due to kneepad use (for knee injury, RR = 0.44, 95% CI: 0.27, 0.74). Knee brace use and ankle brace use were associated with increased rates of knee injury (RR = 1.61, 95% CI: 1.08, 2.41) and ankle injury (RR = 1.74, 95% CI: 1.11, 2.72), respectively. This could be due to slippage of the brace during use, increased fatigue due to the energy cost of wearing a brace, or bias in the study. Further investigation into the effects of brace use is warranted. PMID- 15746468 TI - Risk of functional ovarian cyst: effects of smoking and marijuana use according to body mass index. AB - Smoking is one of the few risk factors that have been identified for functional ovarian cysts, and results of one epidemiologic study suggest that body mass index (BMI; weight (kg)/height (m)(2)) may modify the effect of this exposure. The current study assessed the association of cigarette smoking and marijuana use with functional ovarian cyst risk by using data from a population-based 1990-1995 case-control study of 586 incident functional ovarian cyst cases and 757 age matched controls in a large health maintenance organization in Washington State. In multivariate analyses controlling for age, education, and reference year, the authors found an increase in risk associated with current cigarette smoking among women whose BMI was <20 (odds ratio (OR) = 2.48, 95% confidence interval (CI): 1.32, 4.64) or 20-25 (OR = 1.60, 95% CI: 1.04, 2.46) but not >25 (OR = 0.85, 95% CI: 0.53, 1.37). Corresponding risks associated with current marijuana use were BMI <20, OR = 2.05 (95% CI: 0.89, 4.75); BMI 20-25, OR = 1.78 (95% CI: 1.00, 3.17); and BMI >25, OR = 0.72 (95% CI: 0.36, 1.42). Study results indicate that increased BMI may attenuate the adverse effect of smoking on the risk of functional ovarian cyst. PMID- 15746469 TI - Long-term mobile phone use and brain tumor risk. AB - Handheld mobile phones were introduced in Sweden during the late 1980s. The purpose of this population-based, case-control study was to test the hypothesis that long-term mobile phone use increases the risk of brain tumors. The authors identified all cases aged 20-69 years who were diagnosed with glioma or meningioma during 2000-2002 in certain parts of Sweden. Randomly selected controls were stratified on age, gender, and residential area. Detailed information about mobile phone use was collected from 371 (74%) glioma and 273 (85%) meningioma cases and 674 (71%) controls. For regular mobile phone use, the odds ratio was 0.8 (95% confidence interval: 0.6, 1.0) for glioma and 0.7 (95% confidence interval: 0.5, 0.9) for meningioma. Similar results were found for more than 10 years' duration of mobile phone use. No risk increase was found for ipsilateral phone use for tumors located in the temporal and parietal lobes. Furthermore, the odds ratio did not increase, regardless of tumor histology, type of phone, and amount of use. This study includes a large number of long-term mobile phone users, and the authors conclude that the data do not support the hypothesis that mobile phone use is related to an increased risk of glioma or meningioma. PMID- 15746470 TI - The North Queensland "Sun-Safe Clothing" study: design and baseline results of a randomized trial to determine the effectiveness of sun-protective clothing in preventing melanocytic nevi. AB - In 1999, the authors began recruitment for a randomized controlled intervention trial aimed at preventing melanocytic nevi (moles) by minimizing sun exposure through the use of sun-protective clothing. The study involves 652 Caucasian children (75.6% response) aged 0-35 months from 25 child-care centers (n = 13 intervention and n = 12 control) living in the high-solar-irradiance environment of Townsville, Queensland, Australia. Children attending intervention centers wear investigator-provided garments made from fabrics with ultraviolet protection factors rated very good to excellent. Control centers continue to offer usual care. Three-year follow-up of all children will be completed in 2005. The main outcome measure is the number of new melanocytic nevi. At baseline, the two groups were similar with respect to nevi, phenotype, age, demographic characteristics, sun-protection habits, and history of sun exposure, except that more children from control versus intervention centers (2% and 0%, respectively; p = 0.006) had experienced painful sunburn with blistering. Higher melanocytic nevus counts were associated with more time spent outdoors and a history of sunburn, while sunscreen use, particularly during the mild winter months, appeared to have a protective effect. These findings further substantiate the hypothesis that nevus development in young children is related to sun exposure. PMID- 15746471 TI - Lung function and glucose metabolism: an analysis of data from the Third National Health and Nutrition Examination Survey. AB - Although people with diabetes have decreased lung function, the dose-response relation between measures of glucose control and lung function in nondiabetic people is not known. The authors used data from the Third National Health and Nutrition Examination Survey (1988-1994) to investigate the relation between glucose tolerance test response and other measures of glucose homeostasis and lung function in an adult population without a clinical diagnosis of diabetes. Plasma glucose level 2 hours after oral administration of 75 g of glucose was inversely related to forced expiratory volume in 1 second (FEV(1)), with a difference of -144.7 ml (95% confidence interval: -231.9, -57.4) for persons in the highest quintile of postchallenge glucose compared with the lowest. Similar inverse associations with FEV(1) were found for other measures of glucose autoregulation. Lung function did not appear to be related to fasting glucose level. Similar associations were seen for forced vital capacity (FVC) but not for the FEV(1):FVC ratio. In the total study population, persons with previously diagnosed diabetes had an FEV(1) 119.1 ml (95% confidence interval: -161.5, 76.6) lower than persons without diabetes. This effect was greater in those with poorly controlled diabetes. These findings suggest that impaired glucose autoregulation is associated with impaired lung function. PMID- 15746472 TI - Risk of urinary tract infection and asymptomatic bacteriuria among diabetic and nondiabetic postmenopausal women. AB - No prospective data exist on the risk of microbiologically confirmed urinary tract infection (UTI) and asymptomatic bacteriuria (AB) in relation to diabetes mellitus and its characteristics. The authors prospectively (1998-2002) followed 218 diabetic and 799 nondiabetic Washington State women aged 55-75 years for UTI and AB. The baseline examination and two annual follow-up examinations included urine culture, measurement of hemoglobin A1c and postvoid residual bladder volume, and a survey of diabetes and other characteristics. Surveillance for UTI included self-reports confirmed by microbiologic culture and medical record review. UTI incidence per 100 person-years was 12.2 for diabetic women and 6.7 for nondiabetic women (relative risk (RR) = 1.8, 95% confidence interval (CI): 1.2, 2.7). AB incidence per 100 person-years was 6.7 for diabetic women and 3.0 for nondiabetic women (RR = 2.3, 95% CI: 1.3, 3.9). In Cox models adjusted for multiple covariates, the increased UTI risk occurred mainly in women taking insulin (RR = 3.7, 95% CI: 1.8, 7.3) and women with a longer diabetes duration (> or =10 years; RR = 2.6, 95% CI: 1.3, 5.1) compared with nondiabetic women. No clear linear trend between hemoglobin A1c and UTI or AB risk was seen. Postmenopausal women with diabetes have higher risks of UTI and AB in relation to diabetes duration and severity but not to recent glucose control. PMID- 15746473 TI - Seasonal patterns in monthly hemoglobin A1c values. AB - The purpose of this study was to investigate seasonal variations in population monthly hemoglobin A(1c) (A1c) values over 2 years (from October 1998 to September 2000) among US diabetic veterans. The study cohort included 285,705 veterans with 856,181 A1c tests. The authors calculated the monthly average A1c values for the overall population and for subpopulations defined by age, sex, race, insulin use, and climate regions. A1c values were higher in winter and lower in summer with a difference of 0.22. The proportion of A1c values greater than 9.0% followed a similar seasonal pattern that varied from 17.3% to 25.3%. Seasonal autoregressive models including trigonometric function terms were fit to the monthly average A1c values. There were significant seasonal effects; the seasonal variation was consistent across different subpopulations. Regions with colder winter temperatures had larger winter-summer contrasts than did those with warmer winter temperatures. The seasonal patterns followed trends similar to those of many physiologic markers, cardiovascular and other diabetes outcomes, and mortality. These findings have implications for health-care service research in quality-of-care assessment, epidemiologic studies investigating population trends and risk factors, and clinical trials or program evaluations of treatments or interventions. PMID- 15746474 TI - Transitions between states of disability and independence among older persons. AB - The objectives of this prospective cohort study, conducted in New Haven, Connecticut, from 1998 to 2004, were to describe disability states experienced by older persons, to evaluate the rate of transitions between states and the duration of disability episodes, and to determine whether these findings differ on the basis of physical frailty--a condition of low physical capacity and vulnerability to adverse functional outcomes. Participants included 754 persons aged 70 years or older who were initially independent in four key activities of daily living: bathing, dressing, walking, or transferring. Disability was assessed during monthly telephone interviews for a median of 60 months, and participants were classified each month according to the following four states: no disability, mild disability (one or two activities), severe disability (three or four activities), and death. Transitions between states of disability and independence were common, with a majority of both frail and nonfrail participants experiencing at least one transition. The rate of transitions varied greatly among individuals. Nonfrail participants had lower rates of transition from less to more disability, higher rates of transition from more to less disability, and slightly shorter durations of disability. To fully understand the disabling process, investigators and clinicians must consider the episodic and recurrent nature of disability. PMID- 15746475 TI - Seasonal analyses of air pollution and mortality in 100 US cities. AB - Time series models relating short-term changes in air pollution levels to daily mortality counts typically assume that the effects of air pollution on the log relative rate of mortality do not vary with time. However, these short-term effects might plausibly vary by season. Changes in the sources of air pollution and meteorology can result in changes in characteristics of the air pollution mixture across seasons. The authors developed Bayesian semiparametric hierarchical models for estimating time-varying effects of pollution on mortality in multisite time series studies. The methods were applied to the database of the National Morbidity and Mortality Air Pollution Study, which includes data for 100 US cities, for the period 1987-2000. At the national level, a 10-microg/m(3) increase in particulate matter less than 10 microm in aerodynamic diameter at a 1 day lag was associated with 0.15% (95% posterior interval (PI): -0.08, 0.39), 0.14% (95% PI: -0.14, 0.42), 0.36% (95% PI: 0.11, 0.61), and 0.14% (95% PI: 0.06, 0.34) increases in mortality for winter, spring, summer, and fall, respectively. An analysis by geographic region found a strong seasonal pattern in the Northeast (with a peak in summer) and little seasonal variation in the southern regions of the country. These results provide useful information for understanding particle toxicity and guiding future analyses of particle constituent data. PMID- 15746476 TI - Use of a Bayesian approach to decide when to stop a therapeutic trial: the case of a chemoprophylaxis trial in human immunodeficiency virus infection. AB - From 1996 to 1998, a phase III, placebo-controlled, therapeutic trial was conducted in Abidjan, Ivory Coast, to assess the efficacy of cotrimoxazole prophylaxis in reducing severe morbidity in adults at early stages of human immunodeficiency virus infection. The authors used the real data from this trial to simulate three Bayesian interim analyses. Three prior distributions were considered: a noninformative one, a skeptical one, and one based on external information. The posterior distribution was calculated by using directed acyclic graphs and Gibbs sampling. This Bayesian approach showed different results according to the prior distribution chosen. Although use of the noninformative prior would have led to stopping the trial at the same time that the frequentist approach would have, the skeptical prior would have led to continuing it, and the prior based on external data would have led to stopping it 1 year earlier. PMID- 15746477 TI - Neural underpinnings of fear and its modulation: implications for anxiety disorders. PMID- 15746478 TI - Apathy: why care? AB - This review presents data showing that apathy is common across a number of disorders. Apathy is not only common, but is also associated with significant problems: reduced functional level, decreased response to treatment, poor illness outcome, caregiver distress, and chronicity. Preliminary evidence of treatment efficacy exists for dopaminergic drugs and for amphetamines. Strong evidence of efficacy exists for acetylcholinesterase inhibitors in Alzheimer's disease, and for atypical antipsychotics in schizophrenia. Frontal-subcortical system(s) dysfunction is implicated in the causation of apathy; apathy subtypes based on the various frontal-subcortical loops may thus exist. Further research involving diagnosis, pathophysiology, and treatment is suggested. PMID- 15746479 TI - Mood disorders and their treatment in patients with epilepsy. AB - Mood disorders in patients with epilepsy are not frequently diagnosed and not treated. Because of the high prevalence of depression and the resulting high suicide rate, precise diagnosis and effective therapy are very important. Frequently, the clinical pictures of depressive syndromes in epileptics do not correspond with those described in operationalized classification systems such as ICD-10 or DSM-IV. The incidence of depressive disorders in epileptics is estimated in the literature to be 30%-70%. Multifactorial pathogenetic models include the type of seizures, the location of the epileptic focus, and neurotransmitter dysfunctions, as well as hereditary and psychosocial influences, and negative psychotropic effects of antiepileptic drugs. Despite an insufficient number of available controlled studies, based on the current data, treatment with the newer serotonergic antidepressants can be recommended for patients with epilepsy. PMID- 15746480 TI - The overt aggression scale for rating aggression in outpatient youth with autistic disorder: preliminary findings. AB - Aggression is a common and costly problem in youth with developmental disabilities. Rating scales that accurately capture and measure subtypes of aggression phenomenology, frequency and severity are urgently needed, in both clinical practice and research. The authors studied the Overt Aggression Scale (OAS) in a preliminary sample of eight outpatients who participated in an ongoing placebo-controlled study of valproate for aggression in autism. Subjects' OAS aggression scores showed significant correlation with the already validated retrospectively rated Aberrant Behavior Checklist Community Scale irritability subscale. Further study of the OAS in outpatients with aggression and developmental disabilities is warranted. PMID- 15746481 TI - Clozapine reduces violent behavior in heterogeneous diagnostic groups. AB - Violent behavior is a significant problem in the psychiatric hospital setting. Persistently violent patients often require seclusion and/or restraints and typically receive high doses of medication and polypharmacy. Clozapine has been found to be effective in reducing aggression in patients with psychosis. Thus, we examined the effects of clozapine in a heterogeneous group of persistently violent patients. A chart review of the effect of clozapine in persistently violent patients was performed. Changes in the number of violent episodes and the need for seclusion and restraint were assessed for a 3-month period before and after receiving clozapine. In this group of five, carefully selected, persistently violent patients, clozapine treatment resulted in marked decreases in violent episodes and the use of seclusion and restraint. These data suggest a role for clozapine in the treatment of persistently violent patients irrespective of DSM-IV diagnosis. PMID- 15746482 TI - Characterizing the emotions that trigger cataplexy. AB - Cataplexy is an intriguing example of how emotions can trigger muscle weakness by activating neural pathways. When associated with excessive daytime sleepiness, cataplexy is considered pathognomonic of narcolepsy. A questionnaire was administered to 55 patients with narcolepsy-cataplexy and 47 comparison subjects with obstructive sleep apnea. The area under the receiver-operating curve was 0.94 for the combination of muscle weakness with laughter and ability to hear during the episode. A 51-item questionnaire succeeds in identifying cataplexy in narcolepsy-cataplexy patients measured up against a comparison group. In the future, an abbreviated survey with these two questions should identify cataplexy with high sensitivity and specificity. These selected questions could subsequently be included into screening tools for use with different patient populations. PMID- 15746483 TI - Independence of changes in behavior from cognition and function in community dwelling persons with Alzheimer's disease: a factor analytic approach. AB - The authors' main objective was to investigate the relationship between changes in psychopathological, cognitive and activity of daily living (ADL) instrument scores over 12 months in community-dwelling persons with Alzheimer's disease (AD). A secondary objective was to evaluate the validity of dividing the Clinical Dementia Rating (CDR), a global dementia staging instrument into cognitive and functional subscores. Changes in measures of psychopathology, cognition and function between the baseline and 12-month visits were entered into these post hoc analyses of data from a one-year clinical trial to evaluate behavioral, cognitive and functional assessment instruments for use in clinical trials with AD patients. Exploratory factor analysis was used to determine whether there was independence between changes in any of these three domains of interest for this disease population; participants were a cohort of 187 well-characterized, community-dwelling persons with AD. One-year change in the behavioral symptoms of this cohort of persons with AD was statistically independent from changes in scores on cognitive and functional measures. Some evidence of independence of 12 month changes in cognitive and functional measures was found. Cognitive and functional subscores for the CDR were supported. These findings suggest that changes in behavior and cognition in dementia may have distinct pathophysiologies. PMID- 15746484 TI - Cognitive impairment associated with major depression following mild and moderate traumatic brain injury. AB - Traumatic brain injury (TBI) and major depression are neuropsychiatric conditions that have been associated with cognitive dysfunction. The aim of this study was to explore the relationship between major depression and cognitive impairment following mild and moderate TBI. Seventy-four TBI patients were assessed for the presence of major depression using the Structured Clinical Interview for the DSM IV and completed a neurocognitive assessment battery. Subjects with major depression (28.4%), compared to those without, were found to have significantly lower scores on measures of working memory, processing speed, verbal memory and executive function. Potential mechanisms and implications for treatment are discussed. PMID- 15746485 TI - Deficits in social knowledge following damage to ventromedial prefrontal cortex. AB - Patients with damage to the frontal lobes frequently exhibit impaired social behavior, but it is not clear which specific processes are disrupted. The authors investigated the ability to interpret nonverbal emotional expression in patients with lesions involving ventromedial (N=20) or dorsolateral prefrontal cortex (N=9) and in healthy volunteers (N=23). As hypothesized, only patients with ventromedial prefrontal lesions showed impaired task performance relative to normal comparison subjects. These results suggest that deficits in social knowledge, namely difficulty interpreting nonverbal emotional expression, contribute to the aberrant social behavior observed following ventromedial prefrontal cortex lesions. PMID- 15746486 TI - Increasing memory load modulates regional brain activity in older adults as measured by fMRI. AB - Several recent studies using functional magnetic resonance imaging (fMRI) during recognition memory tests have suggested that the ability to neuromodulate as a function of cognitive demand may be impaired in older adults due to age-related cell loss and neural volume reduction in memory specific regions. In the current study, older adults (ages 59-77) were tested with fMRI during a delayed recognition task in which memory load for faces was varied across trials. Activity was greater in amplitude for three- versus one-face stimuli within the superior, middle, and inferior frontal gyri, intraparietal sulcus, and fusiform gyrus. It was concluded that the ability to modulate activity with increasing load is preserved in older adults despite reductions in neural volume. PMID- 15746487 TI - Normative data for the unquantified EEG: examination of adequacy for neuropsychiatric research. AB - Visual inspection of the unquantified electroencephalogram (EEG) remains the sole technique to confidently detect epileptiform discharges. Usefulness of this procedure in psychiatric research is hampered by reported prevalence of abnormalities in normal adults ranging from 4% to 57.5%. In this study, we examined the criteria used in available literature for selecting healthy individuals. Medline, PsychInfo, and textbook chapters were the primary sources for data. Analysis was restricted to studies examining the visual inspection of the EEG in humans. Thirty-eight articles met search criteria. Seven criteria for choosing healthy comparison subjects were used as the bases for this review. The majority of studies met no criteria, one criterion or two criteria. We conclude that the boundaries for normal unquantified EEG are poorly defined. PMID- 15746488 TI - The neuropsychiatry and multisystem features of the Smith-Magenis syndrome: a review. AB - Smith-Magenis Syndrome (SMS) is a complex, pediatric, neurobehavioral, contiguous gene syndrome ascribed to interstitial microdeletion of chromosome 17, band 11.2. The syndrome is characterized by distinctive behavioral, neurocognitive, and neuropsychiatric abnormalities. This genetically mediated disorder of mental retardation prompts behavioral researchers to examine the links between genes, brain, and behavior in order to solve the gene-behavior puzzle and the genotype/phenotype correlation. In this article, the authors review literature on behavioral profile and its associated psychopathologies, cognitive profiles, multisystem abnormalities, and genetic correlates that highlight the complexities of the disorder. PMID- 15746490 TI - Intellectual functioning in adolescents with indicators of psychosis: evidence for decline in functioning related to number of psychotic features? AB - Substantial research has demonstrated that adults with schizophrenia display intellectual decline compared to their premorbid levels of functioning. Research of this type, however, is not as common in adolescents with psychotic disorders. Since many first-episode adolescents with psychotic disorders other than schizophrenia may eventually meet criteria for this diagnosis, we examined first admission adolescents with variable psychiatric diagnoses. In this study, current intellectual functioning was compared to estimated premorbid functioning (estimated with word recognition reading), and the difference between these scores was related to the number of indicators of psychosis that was present in each case. Subjects consisted of 513 inpatients, ranging in age from 13 to 17 years, who were admitted to the adolescent service of a private psychiatric hospital. Indicators of psychosis came from clinical diagnoses, self-report measures, and clinical rating scales. Across the entire sample of 513 subjects the greater the number of indicators of psychosis that was present, the greater the estimated premorbid/current intelligence quotient (IQ) discrepancy. Type of IQ test, differences in intellectual premorbid functioning, demographic variables, and type of treatment were all unassociated with risk for IQ discrepancy. Within the limitations of estimation of premorbid intellectual functioning, these data suggest that intellectual decline is present at the time of the first psychiatric admission in psychotic adolescent patients who do not necessarily meet diagnostic criteria for schizophrenia and that this discrepancy is greater in patients with more indicators of psychosis. PMID- 15746489 TI - Self-administered screening for mild cognitive impairment: initial validation of a computerized test battery. AB - The Computer-Administered Neuropsychological Screen for Mild Cognitive Impairment (CANS-MCI), a computer administered, scored, and interpreted touch screen battery was evaluated for its ability to detect mild cognitive impairment. Subjects were three hundred ten community-dwelling elders who enrolled in an National Institute on Aging (NIA)-funded study. One-month test-retest reliability correlations were all significant (p<0.05-p<0.001). Concurrent validity correlations were all significant (p<0.001). A high level of diagnostic validity was attained relative to the Weschler Memory Scale-Revised (WMS-R) LMS-II test (p<0.001). Confirmatory factor analysis supported a three-factor model indicating the tests measure the intended cognitive dimensions of memory, language/spatial fluency, and executive function/mental control. Goodness-of-fit indicators were strong (Bentler Comparative Fit Index=0.99; Root Mean Square Error of Approximation=0.055). Initial validation analyses indicate that the CANS-MCI shows promise of being a reliable, valid screening tool in determining whether more intensive testing for early cognitive impairment is warranted. PMID- 15746491 TI - Apathy correlates with cognitive function but not CD4 status in patients with human immunodeficiency virus. AB - Apathy is a prominent neuropsychiatric symptom associated with human immunodeficiency virus (HIV). The increased frequency of apathy in this population may reflect the direct involvement of the virus on the central nervous system (CNS), but the severity of apathy has not been shown to consistently relate to markers of disease activity or other neuropsychiatric complications of the virus. We examined the relationship between ratings of apathy and performance on measures of cognitive function and immune system status in a sample of HIV infected patients. Apathy was significantly elevated among HIV-infected individuals compared to healthy comparison subjects. Apathy was significantly related to performance on measures of learning efficiency and a measure of cognitive flexibility. Ratings of apathy did not relate to CD4 cell count, but they were associated with disease duration. In addition, ratings of depression were independent of ratings of apathy. These findings suggest that apathy does not co-vary with a proxy measure of active disease status, but apathy does relate to several measures of cognitive dysfunction in patients with HIV. As such, the increased prevalence of apathy among HIV-infected adults may reflect HIV associated neurologic dysfunction. PMID- 15746492 TI - Huntington's disease patients show impaired perception of disgust in the gustatory and olfactory modalities. AB - Patients with Huntington's disease show deficits in recognizing disgust in the facial expressions and vocal intonations of others. In this study, the authors demonstrate that these disgust-related deficits extend to foul-smelling olfactory stimuli and inappropriate combinations of taste stimuli. PMID- 15746493 TI - Secondary mania following traumatic brain injury. PMID- 15746494 TI - Of illusions, hallucinations and Creutzfeldt-Jakob disease (Heidenhain's variant). PMID- 15746495 TI - Peripheral circulatory disturbance induced by milnacipran. PMID- 15746496 TI - Measuring the effectiveness of outpatient care. PMID- 15746497 TI - A current perspective on the psychotherapies. PMID- 15746498 TI - Completed suicide in late life. PMID- 15746499 TI - The impact of the "incarceration culture" on reentry for adults with mental illness: a training and group treatment model. PMID- 15746500 TI - SQL Clinic: the open-source alternative for electronic medical records. PMID- 15746501 TI - Defining and measuring clinical effectiveness in the treatment of schizophrenia. AB - OBJECTIVES: Expectations in treating schizophrenia are expanding beyond just controlling psychotic symptoms to include functional recovery. This report describes an approach to define and measure the clinical effectiveness of treatment in achieving these objectives. METHODS: A comprehensive literature review established that there is limited information about the meaning of the term "clinical effectiveness." To address this gap a consensus conference of schizophrenia researchers was held to consider the components of clinical effectiveness in real-world community practice and how these components can best be measured. RESULTS: The consensus of the researchers was that effective clinical treatment is characterized by four outcome domains: symptoms of disease, treatment burden, disease burden, and health and wellness. A clinical instrument to measure these four domains was constructed: Global Outcome Assessment of Life in Schizophrenia (GOALS). In using GOALS, clinicians rate each of the four domains on a scale of 1, very much improved, to 7, very much worse. Field-testing of this instrument is planned. CONCLUSIONS: Effective treatment interventions that combine optimal pharmacotherapy and targeted psychosocial treatments are raising expectations about the prospects of functional recovery among patients with schizophrenia. GOALS is proposed as one tool that can provide busy clinicians with a simple, objective measure of the effectiveness and outcomes of the clinical treatment they provide to patients with schizophrenia. PMID- 15746502 TI - Patterns and quality of treatment for patients with schizophrenia in routine psychiatric practice. AB - OBJECTIVES: This study provided generalizable national data on the treatment of adult patients with schizophrenia in the United States and assessed conformance with the practice guideline treatment recommendations of the Schizophrenia Patient Outcomes Research Team and the American Psychiatric Association. METHODS: National data from the American Psychiatric Institute for Research and Education's 1999 Practice Research Network study of psychiatric patients and treatments were used to examine treatment patterns for 151 adult patients with schizophrenia. Analyses were performed and adjusted for the weights and sample design to generate nationally representative estimates. RESULTS: Findings indicated that patients with schizophrenia who were treated by psychiatrists had complex clinical problems and were markedly disabled. Forty-one percent of patients had a comorbid axis I disorder, and 75 percent were currently unemployed. Thirty-five percent were currently experiencing medication side effects, and 37 percent were currently experiencing problems with treatment adherence. Although most patients received guideline-consistent psychopharmacologic treatment, treatment was characterized by significant polypharmacy. Rates of conformance with the guideline recommendations were significantly lower for psychosocial recommendations than for psychopharmacologic recommendations. Although 69 percent of patients received at least some psychosocial treatment, none of the unemployed patients received vocational rehabilitation services in the past 30 days. CONCLUSIONS: These data suggest unmet need for psychosocial treatment services among individuals with schizophrenia. These findings raise questions about whether currently available antipsychotic medications are being used optimally or whether they offer limited effectiveness for patients with complex clinical problems who are treated in routine psychiatric practice. PMID- 15746503 TI - Construct validity of a measure of subjective satisfaction with life of adults with serious mental illness. AB - OBJECTIVE: In a multisite study, the authors examined the construct validity and utility of a brief self-report Satisfaction With Life scale, an expanded 21-item version of one of the earliest measures of subjective satisfaction with life used with individuals with serious and persistent mental illness. The Satisfaction With Life scale measures satisfaction in four domains: living situation, social relationships, work, and self and present life. METHODS: Satisfaction With Life scale data were gathered from consumers receiving community treatment at two sites in Los Angeles (N=166 and N=172, respectively) and one in Wisconsin (N=146). A confirmatory factor analysis of a hypothesized four-factor structure using data from the pooled Los Angeles samples revealed that several items were less than optimal indicators of the underlying domain. On the basis of an analysis of each item, the Satisfaction With Life scale was reduced to 18 items, and the factor structure and factor loadings of the revised scale were cross validated with data from the Wisconsin sample. The 18-item scale was further validated by testing hypotheses regarding the relationship between the instrument's four domains, or subscales, and clinically important life conditions of clients in the areas of symptoms, living and employment situations, and social relationships. RESULTS: The findings provided excellent support for the construct validity of the 18-item Satisfaction With Life scale, which assesses an individual's subjective satisfaction with his or her current life in the four domains: living situation, social relationships, work, and self and present life. CONCLUSIONS: The brief, easily completed 18-item Satisfaction With Life scale is a useful tool in evaluation research for assessing the subjective satisfaction with life of adults with serious mental illness. PMID- 15746504 TI - A longitudinal perspective on monitoring outcomes of an innovative program. AB - OBJECTIVES: The use of outcome assessment to evaluate the performance of programs over many years of operation is becoming an increasingly important aspect of health care management. Over a five-year period of program monitoring, this study examined changes in individual client outcomes three months after discharge from a residential work therapy program for veterans with severe substance use disorders. The study also examined the relationship between these outcomes and changing program features, such as staffing, treatment variables, and follow-up rates. METHODS: Data on admissions characteristics, services delivered during treatment, and status at discharges were collected for 3,390 veterans who were treated in 25 sites in the Department of Veterans Affairs' Compensated Work Therapy/Transitional Residence program. Follow-up data were gathered three months after discharge for 1,771 veterans (52.2 percent). Hierarchical linear modeling was used to examine the association between year of discharge, site-level measures of program staffing and follow-up rate, and individual patient-level treatment variables and outcomes. RESULTS: Over the five-year monitoring period, site staff-to-bed ratios and follow-up rates dropped substantially, and veterans attended more Alcoholics Anonymous and Narcotics Anonymous meetings and had more toxicology screens. Higher staff-to-bed ratios were associated with more positive employment outcomes, and higher follow-up rates were associated with poorer outcomes in substance abuse and lower total income. However, no significant outcome trends were observed in clinical follow-up measures after the analyses adjusted for these factors, suggesting that program effectiveness did not deteriorate during a period of program change. CONCLUSIONS: Long-term evaluations of program process and follow-up status can usefully document sustained program effectiveness over many years. Such efforts should enhance their value by examining and adjusting for the impact of factors such as changing staffing levels and follow-up rates. PMID- 15746505 TI - Comparison of psychiatrists' and other physicians' assessments of their ability to deliver high-quality care. AB - OBJECTIVE: This study compared psychiatrists' assessments of their ability to deliver high-quality care with those of other physicians. METHODS: Data were used from the Community Tracking Study Physician Survey, a national survey of 12,528 physicians. Linear regression models were used to investigate the effects of type of physician (psychiatrist or other physician), managed care involvement, and ability to obtain inpatient care on four measures of physicians' assessments of quality. RESULTS: In models that did not control for difficulty in obtaining inpatient services, assessments of quality were significantly lower among psychiatrists than among other physicians. Furthermore, although managed care involvement was associated with lower assessments of quality for all physicians in these models, the effects were stronger for psychiatrists than for other physicians. However, after difficulty in obtaining inpatient services was controlled for, psychiatrists' and other physicians' assessments of quality were similar. CONCLUSIONS: Compared with other physicians, psychiatrists' assessments of the quality of care that they provide were lower and their assessments were more influenced by involvement with managed care. These differences may be mediated by difficulty in obtaining inpatient services. When designing and administering behavioral health benefits packages, clinical policy makers should consider the possible effects of reduction in inpatient services on quality of care. PMID- 15746506 TI - Association between community and client characteristics and subjective measures of the quality of housing. AB - OBJECTIVE: The objective of this study was to examine the relationship between subjective perceptions of the quality of housing among mental health consumers and both client characteristics and objective measures of the client's neighborhood. METHODS: A cross-sectional survey of a random sample of 468 male clients who were recruited from three mental health centers in Connecticut was used to assess client characteristics, including sociodemographic and clinical status, and measures of subjective quality of housing that reflected perception of both the client's residence and the neighborhood. Data describing the objective characteristics of the 233 census-tract block-group neighborhoods in which clients lived were obtained from the 2000 decennial census, from which three measures were created by using principal components analysis: average household income, affordability, and availability of plumbing and cooking facilities. Ordinary-least-squares regression analyses were used to identify client and neighborhood correlates of subjective quality of housing. RESULTS: Neither psychiatric diagnosis nor substance abuse were found to be significantly associated with any of the subjective housing quality measures. Clients who were living in their own place with others, those who were less bothered by side effects of medications, and those who were living in higher-income neighborhoods were more satisfied with the overall quality of their housing. CONCLUSIONS: Client assessments of subjective quality of housing, both overall and along specific subdimensions, are largely independent of commonly used diagnostic and symptom measures of mental health status. Consumers' subjective experience of housing quality are significantly associated with objective measures of neighborhood characteristics, particularly the mean household income of the neighborhood. PMID- 15746507 TI - Comparison of clozapine use in Maryland and in Victoria, Australia. AB - OBJECTIVE: Studies of how differences in systems of care, including cultural differences, affect prescribing practice and patient outcomes are important and can help answer questions such as the effectiveness of clozapine in routine practice. This study examined the use of clozapine in Maryland and in Victoria, Australia. METHODS: This study used medical record data to examine the use of clozapine in January 2000 for people with schizophrenia in two different countries. Data were gathered from all six public inpatient facilities in Maryland and from the two main community outpatient centers in Victoria. Outpatients were studied in Victoria because Australia's inpatient mental health facilities have closed and people with treatment-resistant schizophrenia are managed exclusively as outpatients. RESULTS: In Maryland 591 inpatients with schizophrenia were given a prescription for second-generation antipsychotics; in Victoria 356 outpatients with schizophrenia were given such a prescription. Among second-generation antipsychotics, clozapine was used significantly more frequently in Australia than in Maryland for the treatment of schizophrenia (173 prescriptions, or 49 percent, compared with 144 prescriptions, or 19 percent). Both systems used clozapine mostly for the treatment of schizophrenia (94 percent in Victoria compared with 88 percent in Maryland). The mean clozapine dosages that were used for the treatment of schizophrenia were significantly higher in Maryland than in Australia (522 mg per day compared with 431 mg per day). CONCLUSIONS: Significant differences in use and dosages of clozapine were found in two populations that were similar in diagnoses and demographic characteristics. PMID- 15746508 TI - Integrating physical activity into mental health services for persons with serious mental illness. AB - This article reviews evidence supporting the need for interventions to promote physical activity among persons with serious mental illness. Principles of designing effective physical activity interventions are discussed along with ways to adapt such interventions for this population. Individuals with serious mental illness are at high risk of chronic diseases associated with sedentary behavior, including diabetes and cardiovascular disease. The effects of lifestyle modification on chronic disease outcomes are large and consistent across multiple studies. Evidence for the psychological benefits for clinical populations comes from two meta-analyses of outcomes of depressed patients that showed that effects of exercise were similar to those of psychotherapeutic interventions. Exercise can also alleviate secondary symptoms such as low self-esteem and social withdrawal. Although structured group programs can be effective for persons with serious mental illness, especially walking programs, lifestyle changes that focus on accumulation of moderate-intensity activity throughout the day may be most appropriate. Research suggests that exercise is well accepted by people with serious mental illness and is often considered one of the most valued components of treatment. Adherence to physical activity interventions appears comparable to that in the general population. Mental health service providers can provide effective, evidence-based physical activity interventions for individuals with serious mental illness. PMID- 15746509 TI - Diagnosis and treatment of chronic insomnia: a review. AB - OBJECTIVE: Insomnia has high prevalence rates and is associated with significant personal and socioeconomic burden, yet it remains largely underrecognized and inadequately treated. METHODS: A PubMed search for English-language articles covering randomized controlled trials published between 1970 and 2004 was conducted. Search terms used were "insomnia," "behavioral therapy," and the generic names of agents commonly used to treat insomnia (the Food and Drug Administration-approved benzodiazepines and nonbenzodiazepines, trazodone, and over-the-counter agents). RESULTS: Evidence from epidemiologic studies, physician surveys, and clinical studies suggests that numerous patient and physician factors contribute to the fact that the needs of patients with insomnia remain unmet, including low reporting of insomnia by patients, limited physician training, and office-based time constraints, as well as misconceptions about the seriousness of insomnia, the advantages of treatment, and the risks associated with hypnotic use. Nonpharmacologic therapies produce long-lasting and reliable changes among people with chronic insomnia and have minimal side effects. Pharmacologic therapies have proven effective with improving wake time after sleep onset and sleep maintenance and reducing the number of nighttime awakenings. However, pharmacologic therapy has a greater chance of producing side effects. No conclusive evidence exists to favor either pharmacologic therapy or behavioral therapy. CONCLUSIONS: Insomnia is particularly challenging for clinicians because of the lack of guidelines and the small number of studies conducted in patient populations with behavioral and pharmacologic therapies. Current treatment options do not address the needs of difficult-to-treat patients with chronic insomnia, such as the elderly, and those with comorbid medical and psychiatric conditions. More research is necessary to determine the long-term effects of insomnia treatments. PMID- 15746510 TI - Impact of the wait for an initial appointment on the rate of kept appointments at a mental health center. AB - Failure to keep initial appointments at a community mental health center results in a burden on the staff and the center's financial resources. The authors studied referrals to an outpatient program and found that delay in scheduling appointments had a significant impact on rate of kept appointments. The impact was significant during the first week of delay and appeared to stabilize after day seven. Age influenced the rate but differed in influence between the adult and child programs. Changes aimed at reducing wait time for initial appointments may favorably affect rate of kept appointments and ultimately preserve staff and financial resources. PMID- 15746511 TI - Identification and treatment of substance misuse on an inpatient psychiatry unit. AB - This chart review study examined identification and treatment of substance misuse on an inpatient psychiatry ward before and after the hospital's administration made changes to increase attention to patients' substance misuse. Before the ward changes, 113 of 250 inpatients (45 percent) were identified as misusing substances. Misusers were significantly more likely to be younger, male, single, and cigarette smokers. After the ward changes, substance misuse was more than twice as likely to be addressed in treatment planning and discharge diagnoses, and referrals to substance abuse treatment were nearly twice as likely to be made. Changes in assessment of and treatment planning for psychiatric inpatients may increase attention to substance misuse. PMID- 15746512 TI - Housing preferences of homeless veterans with dual diagnoses. AB - Previous research indicates that most homeless persons with mental illness prefer independent living, while most clinicians recommend group housing. This study compared residential preferences of 141 homeless veterans with dual diagnoses with those of 62 homeless nonveterans with dual diagnoses. Clinicians rated both groups while they were in transitional shelters before they were placed in housing. Both samples strongly rejected group home living, but a majority of nonveterans desired staff support. Clinicians recommended staffed group homes for most veterans and nonveterans. This survey underscores the disjuncture between consumers' and clinicians' preferences as well as the need to provide a range of housing options to accommodate varied preferences. PMID- 15746513 TI - Structured exercise for persons with serious psychiatric disabilities. PMID- 15746514 TI - A group-based walking program at a psychiatric rehabilitation center. PMID- 15746515 TI - A lifestyle physical activity program for persons with serious mental illness. PMID- 15746516 TI - Use of CBT in a walking program for veterans with diabetes and depression. PMID- 15746517 TI - Psychologist prescribing. PMID- 15746518 TI - The automobile way of life. PMID- 15746519 TI - Use of child custody as leverage to improve treatment adherence. PMID- 15746520 TI - Psychologist prescribing. PMID- 15746521 TI - Ethical challenges of mental health clinicians in rural and frontier areas. PMID- 15746523 TI - Efforts to encourage entrepreneurship among persons with mental illness. PMID- 15746524 TI - HMOs' restrictions on clinical practice. PMID- 15746526 TI - Mammalian transglutaminases: a family portrait. PMID- 15746527 TI - Structure-function relationships of transglutaminases--a contemporary view. PMID- 15746528 TI - Transglutaminases and their substrates. PMID- 15746529 TI - Type 2 transglutaminase and cell death. PMID- 15746530 TI - Transglutaminase and cell-survival signaling. PMID- 15746531 TI - Transglutaminases in wound healing and inflammation. PMID- 15746532 TI - Transglutaminases in epidermis. PMID- 15746533 TI - Tissue transglutaminase (TG2) in cancer biology. PMID- 15746534 TI - Transglutaminases in neurodegenerative disorders. PMID- 15746535 TI - Tissue transglutaminase and celiac disease. PMID- 15746536 TI - Transglutaminases as biotechnological tools. PMID- 15746537 TI - Extracellular transglutaminase: factor XIII. PMID- 15746538 TI - Transglutaminases of lower organisms. PMID- 15746539 TI - Transglutaminases of higher, lower plants and fungi. PMID- 15746542 TI - Wallenberg syndrome with SIADH. PMID- 15746541 TI - Diabetic nephropathy: the proteinuria hypothesis. AB - BACKGROUND/AIMS: Proteinuria, nearly a universal finding in progressive kidney disease, has been the subject of frequent recent analyses in the renal literature. Proteinuria is a hallmark of diabetic nephropathy: microalbuminuria is the principal early predictor for progression of diabetic glomerulopathy, and proteinuria may be viewed as a measure of the severity and promoter of progression of nephropathy. METHODS: This article critically reviews for the first time the full scope of diabetic proteinuria--complex molecular mechanisms, natural history, and analysis of treatment trials--in order to address the validity of 'the proteinuria hypothesis', i.e., that diabetic proteinuria is a modifiable determinant of renal progression. This hypothesis is analyzed in detail, including recent studies on the primary therapy of diabetic nephropathy, renin-angiotensin blockade. RESULTS: As fully developed, this hypothesis consists of three postulates: that higher amounts of proteinuria predict progressive loss of function, that proteinuria reduction correlates with slowing progression, and that proteinuria is a surrogate endpoint for clinical trials. The latter postulate has not before been adequately linked to growing information about the first two postulates as they apply to diabetic kidney disease. CONCLUSION: While diabetic nephropathy is a disease model for the potential use of proteinuria as a surrogate marker for renal progression, this shift in perspective will require prospective data from additional clinical trials, particularly of non-renin angiotensin blocking drugs, to be complete. PMID- 15746540 TI - Attenuation of interstitial inflammation and fibrosis by recombinant human erythropoietin in chronic cyclosporine nephropathy. AB - BACKGROUND: Evidence suggests that recombinant human erythropoietin (rHuEPO) protects neurons and cardiomyocytes from acute insults. We investigated the protective effect of rHuEPO on cyclosporine (CsA)-induced renal injury. METHODS: CsA (15 mg/kg/day) was given to rats for 1 or 4 weeks, and rHuEPO was concurrently administered at a dose of 100 units/kg (thrice weekly). Effects of rHuEPO on CsA-induced renal injury were evaluated with tubulointerstitial fibrosis (TIF) score, macrophage infiltration, expression of proinflammatory and profibrotic cytokines, and apoptotic cell death. RESULTS: Administration of rHuEPO decreased TIF score and the number of macrophages, which increased significantly in CsA-treated rat kidneys. At the molecular level, rHuEPO treatment decreased proinflammatory mediators (osteopontin and C-reactive protein) and profibrotic mediators (transforming growth factor-beta1 and transforming growth factor-beta1-inducible gene-h3). Increased apoptotic cell death in CsA-treated rat kidneys was significantly decreased with rHuEPO cotreatment, and apoptosis-related genes were regulated in favor of cell survival (increased Bcl-2 and suppressed caspase-3). CONCLUSION: rHuEPO has a renoprotective effect against chronic CsA-induced renal injury. PMID- 15746543 TI - Primary angiitis of the central nervous system: a misinterpreted clinical onset of CNS vasculitis. PMID- 15746544 TI - Diffusion abnormality in Balo's concentric sclerosis: clues for the pathogenesis. PMID- 15746545 TI - Dramatic recovery from prolonged Wernicke-Korsakoff disease. PMID- 15746546 TI - Reversible opsoclonus after diphenhydramine misuse. PMID- 15746547 TI - Intracranial hypotension following chiropraxis. PMID- 15746548 TI - Mary Broadfoot Walker (1888-1974): a historic discovery in myasthenia gravis. AB - As a clinical entity, myasthenia gravis was not recognized until Samuel Wilks in 1877 described bulbar and peripheral muscular weakness. In the Lancet of June 2, 1934, the remarkable discovery of physostigmine treatment by Dr. Mary Walker was published, which was to become the mainstay of symptomatic treatment. A quiet and modest physician, she laboured with considerable success under many obstacles in her work and career. PMID- 15746549 TI - Diplopia. PMID- 15746550 TI - Delayed neurological deterioration in a stroke patient with postoperative acute anemia. PMID- 15746551 TI - Stereotactic Voa-Vop complex thalamotomy for writer's cramp. PMID- 15746552 TI - The value of interleukin-8, interleukin-6 and interleukin-1beta in vaginal wash as predictors of preterm delivery. AB - AIMS: To study the value of interleukin (IL)-8, IL-6 and IL-1beta in vaginal wash as predictors of preterm delivery. METHODS: A prospective analysis of a study group of 200 pregnant women between 24 and 34 weeks of gestation with intact membranes and preterm labor and a control group of 50 pregnant women during the same period of gestation. The controls had uncomplicated pregnancies and subsequently delivered at term. Samples of vaginal secretions were collected from both groups and analyzed for IL-1beta, IL-6 and IL-8 concentrations. RESULTS: Of the women in the study group, 70 had preterm deliveries, while all women in the control group had full-term deliveries. Compared with the control group, the study group had significantly higher concentrations (p < 0.021) of IL-1beta, IL-6 and IL-8. The women in the study group delivering preterm also had IL-1beta, IL-6 and IL-8 concentrations significantly greater (p < 0.001) than those of the same group delivering full term. ROC curves were used to establish cut-off points for the three interleukins to predict preterm delivery. We did not find a cut-off point with an appropriate sensitivity and specificity. CONCLUSION: The concentrations of interleukins in vaginal wash were significantly higher both in the women with preterm labor and in those delivering preterm. While values differed between controls and those with the preterm labor, no cut-off can be obtained to use the results of IL-1beta, IL-6 and IL-8 as a predictor clinically. PMID- 15746553 TI - High-grade neuroendocrine carcinoma of the cervix. A case report. AB - Neuroendocrine tumor of the uterine cervix is a rare and aggressive malignancy. Despite controversial, multimodal treatment methods, prognosis and treatment outcomes are worst in advanced stages. We report an early-stage case treated with the multimodal approach. The nomenclature proposed for this tumor type in 1997 by the College of American Pathologists still has some points of discussion. As in our case some of the tumors cannot be defined exactly into small or large cell types, and this causes confusion at least for nomenclature purposes. A 'mixed' type for this tumor may be appropriate. PMID- 15746554 TI - Immunochemical and biological analysis of allergenicity with excretory-secretory products of anisakis simplex third stage larva. AB - BACKGROUND: Anisakis simplex third stage larvae (L3) are parasites that frequently give rise to allergic responses. The larvae molt into fourth stage larvae (L4), and at each stage they produce L3-excretory-secretory products (L3 ESP) and L4-ESP, respectively, which are different in their main protein constituents. Although the allergenicity of L4-ESP has been investigated by several research groups, research on the allergenicity of L3-ESP has not been carried out by any researcher. In this investigation, the allergenicity and antigenicity of L3-ESP were investigated in comparison with L4-ESP, using rat sera. METHODS: Rat sera were produced by L3 oral infection two times with a 9 week interval. Larvae ESP prepared by culture were concentrated and fractioned using lyophilizer and a centrifugal filter device, respectively. Immunochemical analysis was performed using both indirect ELISA and immunoblot. Biological allergenicity was analyzed by RBL-2H3 exocytosis. RESULTS: With the indirect ELISA, the optical density (OD) value of the nonfractioned (NF)-L3ESP was only one third of that of the NF-L4ESP in both specific IgM and IgG. On measuring specific IgE, the OD of NF-L3ESP was less than one tenth of that of NF-L4ESP. In addition, neither antigen nor allergen was shown in NF-L3ESP, but it was shown in NF-L4ESP with immunoblot. However, the biological allergenicity of NF-L3ESP was comparable to that of NF-L4ESP. To demonstrate the presence of any allergen, L3 ESP was fractioned and found to carry twelve visualized allergen bands from 10 to 186 kDa by immunoblot. CONCLUSIONS: These results indicate that L3-ESP may include the important allergens necessary to induce the allergy by L3 oral infection, as compared to L4-ESP. PMID- 15746555 TI - Clustering of allergic outcomes within families and households in areas endemic for helminth infections. AB - BACKGROUND: Allergy and helminth infections share key immunological features in terms of Th2 responses. Although in industrialized countries clustering of allergic disorders within families has been frequently reported, such information is lacking from areas where helminth infections are endemic. METHODS: A total of 466 subjects from 29 families and 112 households participated in this study. Filarial infection, skin test reactivity and IgE to mite as well as total IgE were measured in all samples. Clustering of the allergy-related outcomes due to genetic and household factors was tested. RESULTS: Genetic factors contributed significantly to the clustering of total IgE and allergen-specific IgE, whereas only household factors contributed to the clustering of SPT positivity. CONCLUSION: Similar to several studies conducted in western populations, total IgE and allergen-specific IgE are influenced by genetic factors in a population resident in a helminth endemic area. However, clustering of SPT positivity due to genetic factors was not significant in the current study raising the question of whether the presence of helminth infections may override genes that are associated with the expression of tissue reactivity to allergens in the west. PMID- 15746556 TI - Increasing prevalence of Japanese cedar pollinosis: a meta-regression analysis. AB - BACKGROUND: Japanese cedar pollinosis, caused by the pollen of the Japanese cedar tree (Cryptomeria japonica), is the commonest seasonal allergic disease in Japan. A number of epidemiological surveys have been reported on Japanese cedar pollinosis, but it has never been assessed systematically or quantitatively. To confirm the increasing prevalence of Japanese cedar pollinosis and related factors, we conducted a meta-regression analysis on population-based surveys in Japan. METHODS: We searched for data from population-based surveys in which serological methods were used to test all participants. Weighted regression of logit-transformed prevalence and sensitization rates were used to evaluate the effects of the year of survey, age, and degree of urbanization. We also analyzed the relationship between prevalence and sensitization rate. RESULTS: Thirty-eight reports with 27 subgroups for prevalence and 134 subgroups for sensitization rate were selected from the literature published in the years between 1986 and 2000. The Japanese cedar pollen sensitization rate was found to be significantly correlated with the year of survey, age, and degree of urbanization (adjusted R(2) = 0.55). The coefficient for the correlation between the prevalence and the sensitization rate revealed a statistically significant correlation (Pearson's r = 0.70, p < 0.001). CONCLUSIONS: The prevalence of Japanese cedar pollinosis among adolescents was predicted to be 28.7% in metropolitan areas and 24.5% in the general population in urban areas in the year 2004, derived from the estimated sensitization rate and the relationship between sensitization rate and prevalence. The prevalence of Japanese cedar pollinosis increased 2.6-fold between 1980 and 2000, and the prevalence differed considerably according to age and degree of urbanization. PMID- 15746557 TI - FK506 aerosol locally inhibits antigen-induced airway inflammation in Guinea pigs. AB - BACKGROUND: Eosinophilic airway inflammation is a common pathological feature of asthma. It has been shown that FK506 given systemically suppresses antigen induced airway inflammation in animal models. However, it is unknown whether inhaled FK506 can suppress the airway allergic inflammation/immune response and whether it acts locally or systemically. METHODS: We tested the effects of oral FK506 and inhaled FK506 on antigen-induced airway inflammation in guinea pigs. The tissue and blood concentrations of FK506 given via both routes were compared. The effect of inhaled FK506 on the expression of cytokine mRNA in lung and bronchoalveolar lavage fluid (BALF) cells was also tested. RESULTS: Both routes of administration of FK506 suppressed the airway eosinophilia in egg albumin (EA) sensitized and -challenged animals. The effect of three inhaled puffs was almost equal to that of 1 mg/kg administered by the oral route. Following inhalation of three puffs, FK506 concentration in blood (AUC(0-24 h)) was approximately 1/21 of that following oral FK506 (1 mg/kg). After EA challenge, mRNA expression of interleukin (IL)-5, eotaxin and IL-1beta in BALF cells and IL-5 in the lung increased significantly. FK506 aerosol markedly inhibited IL-5 mRNA expression in the lung. In situ hybridization indicated that in the BALF IL-5 mRNA expression by lymphocyte-like cells was inhibited by FK506 aerosol. In addition, anti-IL-5 antibody injected intratracheally almost completely abolished eosinophilia in this model. CONCLUSION: Inhaled FK506 can suppress airway inflammation in guinea pigs, where the local action, presumably the inhibition of T-cell activation/function in the lung and airways, was primarily important. PMID- 15746558 TI - Idiopathic CD4+ lymphocytopenia may be due to decreased bone marrow clonogenic capability. AB - BACKGROUND: Idiopathic CD4+ lymphocytopenia is defined by a stable decrease of CD4+ T cells in the absence of any known cause of immune deficiency. The mechanisms responsible for the immunological impairment are still unknown, but a regenerative failure of hematopoietic stem/progenitor cells has been hypothesized. METHODS: We evaluated in the bone marrow (BM) of 5 patients with idiopathic CD4+ lymphocytopenia the phenotype of BM progenitor cells, their differentiation capacity with colony-forming cells and long-term culture initiating cell assays, in parallel with the spontaneous IL-7 production in the patient sera. RESULTS: Compared with controls, a regenerative failure of hematopoietic stem cells has been observed, both in 'committed' and in 'uncommitted' progenitor cells, despite high IL-7 serum levels. The percentage of phenotypically primitive CD34+CD38-DR+ cells (this includes the lymphoid precursor cells) was decreased, suggesting an involvement of the more primitive BM compartment in the de novo T cell generation. CONCLUSIONS: Despite the low number of patients, due to the low incidence of the disease, the decrease of primitive precursors sustains the possibility that diminished stem cell precursors might contribute to the development of CD4+ T cell depletion. PMID- 15746559 TI - Roles of NF-kappaB and bcl-2 in two differential modes of cell death of mouse cortical collecting duct cells. AB - Recent data have implicated nuclear factor-kappaB (NF-kappaB) and Bcl-2 in the regulation of apoptotic and necrotic cell death in various cells. However, mechanisms of their effects on cell death of renal epithelial cells are not clear. First, we investigated the effect of specific inhibition of NF-kappaB and overexpression of Bcl-2 on necrotic cell death induced by hydrogen peroxide or cisplatin in renal collecting duct cells. M-1 cells, which were derived from outer cortical collecting duct, were stably transfected with the non phosphorylatable mutant of inhibitory-kappaBalpha (I-kappaBalpha) and Bcl-2. Overexpression of I-kappaBalpha and Bcl-2 did not affect cisplatin-induced necrotic cell death, but overexpression of I-kappaBalpha significantly decreased H2O2-induced cell death. Regarding apoptotic cell death induced by cisplatin, serum deprivation and contact inhibition was increased by overexpression of I kappaBalpha, whereas overexpression of bcl-2 inhibited the apoptotic cell death. I-kappaBalpha overexpression increased Bax expression and decreased cIAP-1 and -2 expression compared to vector-transfected cells, but did not alter SAPK/JNK activity in the presence or absence of cisplatin. NF-kappaB activity was significantly higher in bcl-2-overexpressing cells than in control cells. These data show that activation of NF-kappaB mediates H2O2-induced necrotic injury, but inhibits apoptotic cell death in renal collecting duct cells, and that Bcl-2 selectively protects apoptotic cell death in M-1 cells. PMID- 15746561 TI - Effect of ex vivo gene transfer with an adenoviral vector on human eye bank corneas. AB - BACKGROUND: Ex vivo gene transfer to donor corneas using adenoviral vectors has gained increasing attention. This study investigates the effect of adenovirus mediated gene transfer on endothelial cell (EC) count in human eye bank corneas. METHODS: A replication-defective adenoviral vector containing the gene for green fluorescent protein was used to transduce organ-cultured normal human eye bank and porcine corneas. Transgene expression and EC count were assessed by light and fluorescence microscopy. RESULTS: The transgene was expressed earlier by porcine EC (27% of all EC on day 2) than by human EC (6% on day 2), but the maximal number of EC finally expressing the transgene was higher in human than in porcine corneas (45 vs. 31% of all EC on day 12). Gene transfer caused considerably less EC loss in human than in porcine corneas (2 vs. 60% EC loss after 10 days). CONCLUSIONS: Adenoviral vectors for ex vivo gene transfer are more efficient and less toxic in normal human eye bank corneas than in porcine corneas, but human EC require more time until expression of transgenic proteins. PMID- 15746563 TI - Translocation of nuclear factor-kappaB on corneal epithelial cells induced by ultraviolet B irradiation. AB - PURPOSE: This study was performed to elucidate the role of nuclear factor-kappaB (NF-kappaB) in the death of corneal epithelial cells after ultraviolet (UV) irradiation. METHODS: Simian virus 40-transfected human corneal epithelial cells (T-HCECs) were used in this study. Cell cultures were irradiated with a UVB (312 nm) source located 10 cm from the bottom of the slides for 10, 20, 30, or 40 s. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide method. Translocation of NF-kappaB was examined by immunocytochemistry using anti-NF-kappaB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine and SN-50, specific NF-kappaB inhibitors, were used to confirm the role of NF-kappaB by pretreating samples for 30 min before UV irradiation, after which cytotoxicity and NF-kappaB translocation were evaluated. RESULTS: When T-HCECs were irradiated with UVB, translocation of NF-kappaB was observed with immunocytochemistry. These translocations peaked 2 h after UV irradiation during EMSA. When pretreated with sulfasalazine or SN-50, the translocation of NF-kappaB was blocked. Cellular death after UV irradiation was also markedly blocked by sulfasalazine. CONCLUSION: These findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation. PMID- 15746562 TI - Effect of steroidal and non-steroidal drugs on the microglia activation pattern and the course of degeneration in the retinal degeneration slow mouse. AB - BACKGROUND: In hereditary retinal degeneration, microglia cells become activated, migrate through the outer nuclear layer (ONL) and accumulate in the subretinal space. Although this inflammatory process is not likely to be responsible for the onset of photoreceptor apoptosis, cytotoxic substances secreted by activated microglia could potentially accelerate and perpetuate the degenerative process. Anti-inflammatory drugs have been shown to modulate the microglia response in neurodegenerative disorders and potentially ameliorate the disease progression in various animal model systems. In this study we wanted to test the impact of the most commonly used anti-inflammatory drugs (acetylsalicylate and prednisolone) on the microglia activation pattern, the rate of caspase-3-dependent photoreceptor apoptosis and the course of the degeneration in the retinal degeneration slow (rds) mouse retina. METHODS: 169 pigmented rds mice and 30 CBA wild-type mice were used for this study. The treatment groups were injected daily with either acetylsalicylate (200 mg/kg) or prednisolone (2 mg/kg) i.p. from day 0 up to 3 months. Animals were sacrificed at days 10, 14, 16, 18, 20, 30, 40, 60 and 90. Cryoprotected frozen sections were immunostained with F4/80 and cleaved caspase-3 antibodies. The main outcome measures were the total microglia count in the subretinal space, the total cleaved caspase-3-positive cells in the ONL and the averaged number of photoreceptor rows in the midperipheral retina. RESULTS: Neither acetylsalicylate nor prednisolone reduced subretinal microglia accumulation in the rds mouse degeneration model. Moreover, they aggravated migration and accumulation in the early time course. The apoptotic cascade started earlier and was more pronounced in both treatment groups compared to the control group. The pace of retinal degeneration was not reduced in the treatment groups compared to the untreated control. In contrast, acetylsalicylate did significantly accelerate the photoreceptor cell degeneration in comparison to the prednisolone (p < 0.001) and to the control group (p < 0.001). CONCLUSIONS: Acetylsalicylate and prednisolone do not decrease the microglia response in the rds mouse and are not neuroprotective. More research is needed to clarify the molecular mechanisms which lead to photoreceptor cell death and to elucidate the complex role of microglia in inherited retinal degeneration. PMID- 15746564 TI - Molecular analysis of RIM1 in autosomal recessive Retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a frequent retinal dystrophy characterized by a progressive loss of photoreceptors along with retinal degeneration. RIM1, encoding a presynaptic protein involved in the glutamate neurotransmission, is the responsible gene for autosomal dominant cone-rod dystrophy CORD7, whose locus overlaps partially with a locus of autosomal recessive RP (arRP), RP25. Given the genetic heterogeneity that features RP, it is plausible that mutations in RIM1 are also implicated in the disease in arRP families genetically linked to the CORD7 region. To test our hypothesis we analysed the complete RIM1 gene in 8 arRP families by DNA sequencing. Even though the absence of pathogenic mutations suggests that RIM1 is notinvolved in arRP, a role for this gene in other inherited forms of RP as well as other retinal dystrophies needs to be elucidated. PMID- 15746565 TI - Change of proliferation rate of corneal epithelium in the rabbit with orthokeratology lens. AB - OBJECTIVE: To investigate the cell proliferation rate of normal corneal epithelium with extended orthokeratology lens (OKL) wear in comparison with extended rigid gas-permeable (RGP) lens wear. METHODS: Twenty-four rabbits were fitted unilaterally with either an OKL or an RGP lens, and the other eye was used as a control. They were injected with 5-bromo-2-deoxyuridine (BrdU) 24 h prior to being sacrificed. The rabbits were sacrificed at 1, 3, 7 and 14 days after lens fitting. The cornea from the superior limbus to the center was taken at 1.0-mm intervals. The BrdU-labeled cells were counted in medium power fields (x200) in each sample using light microscopy. RESULTS: The number of BrdU-labeled cells in the RGP lens group initially increased, but the number decreased in the corneal center and superior limbus by 32 and 8%, respectively, after 14 days. There was no statistically significant change. However, the number of BrdU-labeled cells in the OKL group decreased after 3 days, and the number of BrdU-labeled cells was reduced in the center and superior limbus by 63 and 8%, respectively, after 14 days. The change in proliferation in the corneal center in the OKL-wearing rabbits was statistically significant compared to the control (p < 0.05). CONCLUSIONS: Wearing an OKL had a greater effect on the change of the proliferation pattern in the epithelium than wearing an RGP lens, which suggests that the OKL might be less physiologic than the RGP lens is. PMID- 15746566 TI - Development of the embryonic porcine neuroretina in vitro. AB - PURPOSE: The objective of this study was to investigate the survival and morphology of embryonic porcine full-thickness neuroretina in culture. METHODS: Porcine fetuses were taken out by cesarian section, and the eyes were enucleated. Neuroretinas were explanted on culture plate inserts and were kept for 0-42 days in vitro under standard culture conditions. Green nucleic acid (Sytox) was used for measuring the extent of cell death, and 4,6-diaminidine-2 phenylindoldihydrochloride was used as a marker for the cellular layers. The explants were examined as whole-mount preparations and vertical sections. Sectioned tissue was stained with hematoxylin-eosin and labeled for immunohistochemistry with photoreceptor-specific antibodies raised against transducin and recoverin. RESULTS: In explants kept for 0-5 days in vitro, the developing retina consisted of multiple rows of neuroblastic cells and a more defined, but multilayered ganglion cell layer (GCL). Older explants revealed a more differentiated appearance with ultimately all normal retinal layers present, even after 42 days in vitro. Transducin- and recoverin-labeled photoreceptors were seen in these specimens, but no outer segments were found. The whole-mount preparation revealed extensively Sytox-labeled cells in the GCL at 2 days in vitro, but very few cells were labeled in older explants. CONCLUSION: This study shows that cultured fetal porcine full-thickness neuroretina can survive and develop according to its intrinsic timetable for at least 6 weeks in vitro. The in vitro system for culturing of the full-thickness retina may be useful in experiments involving retinal transplantation. PMID- 15746567 TI - Serum cytokine receptor levels in noninfectious uveitis. AB - PURPOSE: Understanding of the role of cytokines in uveitis may provide new clues to its treatment. Therefore, the purpose of our study was to evaluate systemic cytokine receptor expression in patients with noninfectious uveitis. METHOD: Serum concentrations of soluble interleukin-2 receptor alpha (IL-2 s Ralpha) and soluble tumor necrosis factor receptor-1 (sTNF-R1) were measured in patients with intermediate uveitis (n = 26), posterior uveitis (n = 23) and healthy controls (n = 12) using ELISA. All patients were identified in a consecutive series of 996 uveitis patients who had been diagnosed between 1998 and 2002 and classified according to the recommendations of the International Uveitis Study Group. Inclusion criteria were idiopathic, active intraocular inflammation, uveitis as a primary process and no systemic anti-inflammatory treatment at the time of blood sampling. None of the patients had an underlying systemic disease. RESULTS: Serum concentrations of IL-2 s Ralpha were significantly increased in patients with posterior (p < 0.005) and intermediate uveitis (p < 0.005) as compared to healthy controls. Similarly, concentrations of sTNF-R1 appeared to be increased in posterior (p < 0.005) and intermediate (p < 0.005) uveitis patients when compared to controls. CONCLUSIONS: Our results may suggest that patients with noninfectious uveitis express systemic cytokine receptors such as TNF-R1 and IL-2 Ralpha, which may have an important role in the immune response of the eye and may lead to new immunomodulatory approaches. PMID- 15746568 TI - Comparison of in vitro susceptibilities of ocular bacterial isolates to gatifloxacin and other topical antibiotics. AB - PURPOSE: To compare the in vitro activity of gatifloxacin with that of other topical antibiotics against fresh bacterial isolates from various types of human ocular infections. METHODS: A total of 498 strains of ocular bacterial isolates were tested for their susceptibility to gatifloxacin, ofloxacin, ciprofloxacin, norfloxacin, gentamicin, tobramycin, neomycin, chloramphenicol, erythromycin, tetracycline and amikacin. RESULTS: Ninety-nine percent of gram-positive and 92% of gram-negative bacterial strains were found to be susceptible in vitro to gatifloxacin, whereas lower percentages of gram-positive and gram-negative bacteria were found to be susceptible to other antibiotics. CONCLUSIONS: In vitro, gatifloxacin exhibits good activity against various bacterial species isolated from different ocular sites. This requires corroboration in the clinical setting. PMID- 15746569 TI - Laboratory diagnosis in ulcerative keratitis. AB - AIMS: To identify the common bacterial and fungal isolates from corneal ulcers and to determine the antimicrobial susceptibility patterns of bacterial isolates to commonly used antibiotics at B.P. Koirala Institute of Health Sciences (BPKIHS), eastern Nepal. Culture and direct microscopic correlation and reliability were also compared. METHODS: All patients with suspected corneal ulceration presenting to the Ophthalmology Department of BPKIHS from 1st August 1998 to 31st July 2001 were evaluated. Corneal scraping was performed and processed for direct microscopy and culture for bacterial and fungal isolates. Bacterial isolates were subjected to antimicrobial susceptibility testing. RESULTS: Of 447 specimens examined direct microscopy was positive in 216 (48%) specimens. Culture positivity could be correlated with direct microscopy in 179 (83%) of specimens. Growth of etiologic agents was found in 303 (67.8%) samples. Of these 145 (47.8%) had pure fungal growth, 103 (34%) had pure bacterial growth and 55 (18.2%) had mixed fungal and bacterial infection. The commonest fungal pathogen was Aspergillus spp.in 78 (38.4%) followed by Fusarium spp. in 45 (22%). Aureobasidium sp. was isolated in 25 (12.3%) samples. Staphylococcus aureus (93, 56.7%) dominated the scene as the commonest bacterial agent. Streptococcus pneumoniae (33, 20%) was second in the list. Most of the bacterial isolates were sensitive to commonly used antibiotics. CONCLUSION: This study emphasizes the importance and need of the continued surveillance of the agents and their antimicrobial susceptibility for the prevention and management of corneal ulcers and their complications. PMID- 15746570 TI - Effect of glucagon-like peptide-1(7-36) and exendin-4 on the vascular reactivity in streptozotocin/nicotinamide-induced diabetic rats. AB - We investigated the vascular effects of glucagon-like peptide-1 (GLP-1) and Exendin-4 in type 2 diabetic rat aortae. Studies were performed in a normal control group (NC) (0.2 ml i.p. saline, n = 10), streptozotocin (STZ)/nicotinamide diabetic control group (DC) (a single dose of 80 mg/kg STZ i.p. injection 15 min after administration of 230 mg/kg nicotinamide i.p.), GLP-1 (GLPC) control group (1 microg/kg twice daily i.p. for 1 month, n = 10), Exendin 4 control group (EXC) (0.1 microg/kg twice daily i.p. for 1 month, n = 10), GLP-1 treated diabetic group (GLPT) (1 microg/kg twice daily i.p. for 1 month, n = 10), and Exendin-4-treated diabetic group (EXT) (0.1 microg/kg twice daily i.p. for 1 month, n = 10). One month of GLP-1 and Exendin-4 treatment significantly decreased the blood glucose levels of diabetic rats (113 +/- 2 mg/dl, p < 0.001, and 117 +/- 1 mg/dl, p < 0.001, respectively versus 181 +/- 9 mg/dl in the DC group). Sensitivity (pD2) and maximum response (% Max. Relax) of acetylcholine stimulated relaxations in the DC group (pD2: 6.73 +/- 0.12 and 55 +/- 6, respectively) were decreased compared with the non-diabetic NC group (pD2: 7.41 +/- 0.25, p < 0.05, and 87 +/- 4, p < 0.01). Treating diabetic rats with GLP-1, pD2 values and with Exendin-4, Max. Relax %values of aortic strips to acetylcholine returned to near non-diabetic NC values (pD2: 7.47 +/- 0.15, p < 0.05, and 87 +/- 3, p < 0.01, respectively). Maximal contractile responses (Emax) to noradrenaline in aortic strips from the diabetic DC group (341 +/- 27 mg tension/mg wet weight) were significantly decreased compared with the non diabetic NC (540 +/- 66 mg tension/mg wet weight, p < 0.001) and the GLPT group (490 +/- 25 mg tension/mg wet weight, p < 0.05). There were no significant differences in pD2 values of aortic strips to noradrenaline from all groups. Emax to KCl in aortic strips from the DC group (247 +/- 10 mg tension/mg wet weight, p < 0.01) was significantly decreased compared with non-diabetic NC group (327 +/- 26 mg tension/mg wet weight). Treating diabetic rats with GLP-1 (GLPT), Emax values of aortic strips to KCl returned to near non-diabetic NC values (271 +/- 12 mg tension/mg wet weight). GLP-1 and (partially) Exendin-4 treatment could improve the increased blood glucose level and normalize the altered vascular tone in type 2 diabetic rats. PMID- 15746571 TI - From bench to bedside for gene-directed enzyme prodrug therapy of cancer. AB - Gene therapy of cancer offers the possibility of a targeted treatment that destroys tumors and metastases, but not normal tissues. In gene-directed enzyme prodrug therapy (GDEPT), or suicide gene therapy, the gene encoding an enzyme is delivered to tumor cells, followed by administration of a prodrug, which is converted locally to a cytotoxin by the enzyme. The producer cells as well as surrounding bystanders are subsequently killed. Promising results have meant that suicide gene therapy has reached multicenter phase III clinical trials. This review will discuss the development, efficiency, mode of action and pharmacokinetics of seven GDEPT systems in vitro and in vivo. We will review the latest data of those systems in clinical trials (herpes simplex virus thymidine kinase/gancyclovir, bacterial cytosine deaminase/5-fluorocytosine, bacterial nitroreductase/CB1954 and cytochrome P450/cyclophosphamide), as well as the development of more recent and experimental systems which are not yet in clinical trials (P450 reductase/tirapazamine, carboxypeptidase/CMDA, horseradish peroxidase/indole-3-acetic acid or paracetamol and others). PMID- 15746572 TI - Chemotherapy strategies in the treatment of small cell lung cancer. AB - Lung cancer is the most prevalent, yet most preventable malignancy worldwide. Given the tendency of small cell lung cancer (SCLC) to early relapse and its subsequent resistance to treatment, there is an urgent need to optimize standard treatment strategies and develop new treatments. Over the last decade, several strategies have been adopted and advances in the molecular biology of lung cancer have identified a number of targets for future therapy. In this article, we review chemotherapy strategies that have been evaluated in the management of patients with SCLC. PMID- 15746573 TI - Hyaluronic acid butyric esters in cancer therapy. AB - In this review we focus on a promising novel histone deacetylase (HDAC) inhibitor (HA-But) obtained by the esterification of butyric acid (BA), the smallest HDAC inhibitor, with hyaluronic acid (HA), the main constituent of the extracellular matrix which selectively recognizes a transmembrane receptor (CD44) overexpressed in most primary cancers and associated with tumor progression. In vitro, HA-But has proved to be 10-fold more effective than BA in inhibiting the proliferation of a panel of human cancer cell lines, representative of the most common human cancers, and, similar to BA, to regulate the expression of some cell cycle related proteins, to induce growth arrest in the G1/G0 phase of the cell cycle and to increase histone acetylation. In vivo, HA-But treatment has demonstrated a marked potency in inhibiting primary tumor growth and lung metastases formation from murine Lewis lung carcinoma (LL3) as well as liver metastases formation from intrasplenic implantation of LL3 or B16-F10 murine melanoma cells. In particular, the effect of s.c. and i.p. treatment with HA-But on liver metastases resulted, respectively, in 87 and 100% metastases-free animals, and in a significant prolongation of the survival time compared to the control groups. The results suggest that the presence of the HA backbone does not interfere with the biological activity of butyric residues and that HA-But could represent a promising cell-targetable antineoplastic agent for the treatment of primary and metastatic tumors. PMID- 15746574 TI - Cytotoxicity of RH1: NAD(P)H:quinone acceptor oxidoreductase (NQO1)-independent oxidative stress and apoptosis induction. AB - The elevated expression of the flavoprotein NAD(P)H:quinone acceptor oxidoreductase (NQO1) (EC 1.6.99.2) in many human solid tumors, along with its ability to activate quinone-based anticancer agents, makes it an excellent target for enzyme-directed drug development. Previous studies have shown a significant statistical correlation between NQO1 enzymatic activity and the cytotoxicity of certain antitumor quinones. RH1 [2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4 benzoquinone], presently in late preclinical and entering early clinical development, has been previously considered to be an excellent substrate for activation by NQO1. In this study we investigate the cytotoxicity of RH1 in cell lines selected from the NCI's 60 tumor cell line panel, expressing varying levels of NQO1 activity. Exposure time- and concentration-dependent cytotoxicity was seen, apparently independent from levels of NQO1 activity in these cells. Furthermore, the NQO1 inhibitor dicoumarol had no impact on the sensitivity profiles of RH1 response. The HL-60 myeloid leukemia cells, which do not have detectable NQO1 activity, were further investigated. RH1 treatment of HL-60 cells generated high levels of free radicals, which was accompanied by robust redox cycling, oxygen consumption and induction of apoptosis. These results are in agreement with previous data suggesting that, in addition to its activation by NQO1, RH1-induced cytotoxicity might involve alternative pathways for activation of this compound. Furthermore, the high cytotoxicity of RH1 in the leukemia/lymphoma subpanel of the NCI in vitro cell line screen would suggest an empirical rationale for the utilization of this compound in the treatment of these malignancies. PMID- 15746575 TI - High cytotoxic sensitivity of the human small cell lung doxorubicin-resistant carcinoma (GLC4/ADR) cell line to prodigiosin through apoptosis activation. AB - In the present study, we describe the cytotoxicity of the new drug prodigiosin (PG) in two small cell lung carcinoma (SCLC) cell lines, GLC4 and its derived doxorubicin-resistant GLC4/ADR cell line, which overexpresses multidrug-related protein 1 (MRP-1). We observed through Western blot that PG mediated cytochrome c release, caspase cascade activation and PARP cleavage, thereby leading to apoptosis in a dose-response manner. MRP-1 expression increased after PG treatment, although that does not lead to protein accumulation. The MTT assay showed no difference in sensitivity to PG between the two cell lines. Our results support PG as a potential drug for the treatment of lung cancer as it overcomes the multidrug resistance phenotype produced by MRP-1 overexpression. PMID- 15746576 TI - Anti-angiogenic properties of plaunotol. AB - We have investigated a potential anti-angiogenic effect of plaunotol, an extract from the leaves of Plau-noi, in an angiogenesis model consisting of human umbilical vein endothelial cells (HUVECs). Plaunotol inhibited the proliferative activity of HUVECs in a dose-dependent manner. In addition, it caused a remarkable decrease of the ability of HUVECs to adhere and spread on gelatin and vitronectin, but not fibronectin. Tube-like formation in Matrigel was also inhibited in a dose-dependent way. These results strongly suggest the specific inhibition of integrin alphavbeta3 to be the main event of plaunotol-induced suppression of angiogenesis. The alphavbeta3 antagonists are known to be potent inhibitors of tumor angiogenesis and plaunotol, by causing the functional inhibition of alphavbeta3, should be considered a promising new anti-angiogenic drug. PMID- 15746577 TI - Induction of cell cycle arrest and apoptosis in human non-small cell lung cancer A549 cells by casuarinin from the bark of Terminalia arjuna Linn. AB - Casuarinin, a hydrolyzable tannin isolated from the bark of Terminalia arjuna Linn. (Combretaceae), inhibits human non-small cell lung cancer A549 cells by blocking cell cycle progression in the G0/G1 phase and inducing apoptosis. Enzyme linked immunosorbent assay showed that the G0/G1 phase arrest is due to p53 dependent induction of p21/WAF1. An enhancement in Fas/APO-1 and the two forms of Fas ligand (FasL), membrane-bound FasL and soluble FasL, might be responsible for the apoptotic effect induced by casuarinin. Our study reports here for the first time that the induction of p53 and the activity of the Fas/FasL apoptotic system may participate in the antiproliferative activity of casuarinin in A549 cells. PMID- 15746578 TI - In vitro assessment of cytotoxic agent combinations for hormone-refractory prostate cancer treatment. AB - We have investigated new drug combinations of potential clinical value for treatment of hormone-refractory prostate cancer. Combinations of paclitaxel, carboplatin and mitoxantrone, and combinations of these three drugs with compounds targeting important pathways for cancer progression, 13-cis-retinoic acid and chelerythrine, were assessed. The drugs combinations were incubated for 72 h in steroid-free conditions with two androgen-independent cell lines, DU145 and PC3. Cytotoxicity assay was performed using resazurin and Hoescht 33342. Synergism and antagonism were measured by the combination index, and calculated for each combination by the median-effect method. All six compounds exhibited cytotoxic effects when tested alone. Paclitaxel exhibited the highest and 13-cis retinoic acid the lowest effect on both cell lines. Paclitaxel demonstrated synergism or additivity with 13-cis-retinoic acid in both cell lines, whereas antagonistic effects were observed when it was tested in combination with carboplatin. Chelerythrine showed additive effects with mitoxantrone in both cell lines and with paclitaxel in PC3 cells. Our results suggest that combination of paclitaxel and 13-cis-retinoic acid, and of chelerythrine with mitoxantrone and paclitaxel, may have clinical value for the treatment of hormone-refractory prostate cancer. PMID- 15746579 TI - In vivo efficacy and toxicity of intratumorally delivered mitomycin C and its combination with doxorubicin using microsphere formulations. AB - The efficacy and toxicity of intratumorally (i.t.) administered anticancer drugs mitomycin C (MMC) and doxorubicin (Dox) incorporated in polymeric microspheres were investigated. Biodegradable sulfopropyl dextran microspheres and their oxidized products were used to load Dox and MMC, respectively. EMT6 mouse mammary cancer cells were injected into the hind leg of BALB/c mice. MMC microspheres, alone or combined with Dox microspheres, were injected i.t. once tumors had reached around 0.3 g. The tumor-plus-leg diameter was measured daily and the delay in time for the tumor to grow to 1.13 g relative to control (TGD) was employed as an indication of therapeutic effect. General toxicity was determined by monitoring weight, appearance and behavior of the mice. Morphology and histology of tumor and heart tissues were also examined. An average 79% TGD was observed after i.t. injection of MMC microspheres. The i.t. co-administration of MMC and Dox microspheres resulted in a 185% TGD. The i.t. injections of the microsphere formulations did not result in visible signs of toxicity in animals. In contrast, systemic (i.e. i.p.) injections of MMC solutions caused considerable general toxicity. This study suggests that i.t. delivery of anticancer drugs by polymeric microspheres is an effective way of improving the therapeutic index for cancer chemotherapy of selected solid tumors under special conditions. PMID- 15746580 TI - Pegylated liposomal doxorubicin and mitomycin C in combination with infusional 5 fluorouracil and sodium folinic acid in the treatment of advanced gastric cancer: results of a phase II trial. AB - Mitomycin C (MMC) in combination with infusional 5-fluorouracil (5-FU) is a well tolerated active combination therapy for advanced gastric cancer. Pegylated liposomal doxorubicin (Caelyx) has been combined with this regimen in a phase I study exhibiting promising activity in patients with upper gastrointestinal tumors. In the present study, we investigated activity and tolerability of this three-drug regimen in patients with gastric cancer. Patients with advanced or metastatic gastric cancer were recruited to receive weekly infusional 5-FU (2000 mg/m2) mixed with sodium folinic acid (FA; 500 mg/m2) in one pump (days 1, 8, 15, 22, 29, 36). On days 1 and 29, Caelyx (20 mg/m2) was given as a 1-h, and MMC (7 mg/m2) was applied as bolus injection on days 8 and 36. Treatment courses were repeated on day 57. Twenty-seven patients with a median age of 66 years were recruited in a single center; 56% had histologically proven peritoneal carcinomatosis and 26 patients are evaluable for toxicity. Common Toxicity Criteria of the National Cancer Institute grade 3 toxicity was recorded in 34% of the patients (anemia 12%, leukocytopenia 8%, febrile neutropenia 4%, thrombocytopenia 12%, nausea 15%, diarrhea 8% and mucositis 4%). One patient developed hemolytic-uremic syndrome. One complete (5%) and eight partial responses (42%) were observed in 19 patients evaluable for response according to WHO criteria. Seven patients had no change (37%) and three (16%) progressive disease. Six patients with peritoneal carcinomatosis not amenable to WHO response assessment had progression-free intervals between 8 and 21 months. Median survival for all patients was 14.7 months and median time to progression was 8.4 months. We conclude that this new three-drug combination regimen yields a promising overall response rate (47%) in patients with gastric cancer despite the inclusion of a majority of elderly patients at moderate or high risk of death in this trial. Its safety and good tolerability as established in the phase I trial was confirmed. PMID- 15746581 TI - Preoperative chemotherapy with epidoxorubicin, docetaxel and capecitabine plus pegfilgrastim in patients with primary breast cancer. AB - The objective of this pilot trial was to evaluate the safety and activity profile of epidoxorubicin, docetaxel and oral capecitabine plus pegfilgrastim (TEX+P) as preoperative first-line treatment for patients with breast cancer. Eleven consecutive patients were enrolled in this prospective clinical pilot trial. Preoperative treatment consisted of epidoxorubicin [75 mg/m2 body surface area (BSA)] and docetaxel (75 mg/m2 BSA) administered sequentially on day 1 in combination with oral capecitabine 2000 mg/m2 daily divided into two doses on days 1-14 of each 3-week treatment cycle. Pegfilgrastim 6 mg fixed dose was administered s.c. on day 2 of every treatment cycle. Patients received a total of 58 cycles (median 6 cycles, range 1-6) of this therapeutic regimen. Outpatient TEX+P was well tolerated. No WHO grade IV toxicity was observed. A pathological major response to this preoperative therapy regimen could be demonstrated in eight of nine evaluable patients leading to breast-conserving surgery in seven of nine evaluable patients. We conclude that outpatient TEX+P is safe in the neoadjuvant treatment of patients with primary breast cancer. Thus, this regimen can be considered for further clinical trials. PMID- 15746582 TI - Phase II trial of cisplatin, tegafur plus uracil and leucovorin as neoadjuvant chemotherapy in patients with squamous cell carcinoma of the oropharynx and hypopharynx. AB - We evaluated the efficacy and toxicity of cisplatin, tegafur plus uracil and leucovorin as neoadjuvant chemotherapy for locally advanced squamous cell carcinoma (SCC) of the oropharynx and hypopharynx. Forty-six patients (stage IV, 83%; N2/3, 52%) were treated with PUL (50 mg/m2 cisplatin on day 1, 300 mg/m2 tegafur plus uracil orally and 60 mg leucovorin orally on days 1-14) over a 14 day cycle. Evaluation after 3 cycles led to chemotherapy termination if primary tumor responses were less than partial responses. Otherwise, PUL was continued up to 6 cycles before locoregional therapy. Patients achieving at least good partial responses at the primary site after neoadjuvant chemotherapy received radiotherapy for organ preservation. Chemotherapy responses were analyzed by intent-to-treat. Response rates of primary sites were 71.7% (33 of 46) with 34.8% (16 of 46) showing a complete response. Thirty patients (65.2%) achieved good partial responses at the primary site. Overall response and complete response rates of neck lymph nodes were 68.6% (24 of 35) and 25.7% (nine of 35). The combined response rate of primary site and neck lymph nodes was 63% (95% confidence interval 48.5-77.5%) with a complete response rate of 15.2%. Toxicities of WHO grade 3-4 included anemia (19.6%), diarrhea (17.4%) and neutropenia (8.7%). With a median follow-up of 36 months, overall survival and disease-free survival rates were 45.7% (21 of 46) and 41.3% (19 of 46); organ preservation rate was 90% (19 of 21). We concluded that the outpatient PUL regimen was a moderately effective, less-toxic neoadjuvant chemotherapy for SCC of the oropharynx and hypopharynx. PUL should be studied further with other active agents or radiotherapy. PMID- 15746583 TI - Phase II study of weekly docetaxel and cisplatin in patients with non-small cell lung cancer. AB - We conducted a phase II study to examine the efficacy and safety of weekly docetaxel and cisplatin in the treatment of advanced non-small cell lung cancer (NSCLC). Forty chemotherapy-naive patients (10 with stage IIIB and 30 with stage IV NSCLC) with an Eastern Cooperative Oncology Group performance status of 0-2 and adequate organ functions were enrolled. Chemotherapy consisted of cisplatin (80 mg/m2) on day 1, and docetaxel (35 mg/m2) on days 1, 8 and 15, delivered in 4 week cycles consisting of three weekly treatments followed by 1 week of rest. There were 18 partial responses, with an overall response rate of 45% (95% confidence interval 29.6-60.4%) in 40 treated patients. The median survival period was 19.9 months, median progression-free survival was 5.5 months and 1 year survival rate was 69.4%. Hematologic toxicities were mild and included grade 3 or 4 neutropenia in 37.5%. There were no severe infections or septic deaths. Non-hematologic toxicities were generally mild. Grade 3 or 4 transaminase elevations were observed in two patients. Grade 3 events included two cases each of vomiting, and one case each of hypokalemia, diarrhea and creatinine elevations. Weekly docetaxel and cisplatin is an effective and safe combination in the treatment of patients with advanced NSCLC. PMID- 15746584 TI - Sustained complete remission of metastatic dermatofibrosarcoma protuberans with imatinib mesylate. AB - Dermatofibrosarcoma protuberans (DFSP) is a soft tissue tumor which may recur locally and rarely causes metastases to vital organs. DFSPs have specific chromosomal abnormalities involving the platelet-derived growth factor beta-chain locus (PDGFB) which may render these tumors responsive to targeted therapy with the tyrosine kinase inhibitor imatinib mesylate. A patient with locally recurrent and metastatic DFSP resistant to first-line chemotherapy was treated with imatinib mesylate 400 mg/day. The tumor was examined by a novel fluorescence in situ hybridization (FISH) method for specific rearrangements of the PDGFB locus. The patient was followed for response and toxicity by physical examination and imaging studies. FISH revealed PDGFB rearrangement indicative of multiplication of the PDGFB fusion locus within a ring chromosome. Physical examination showed response within the first month of treatment, and subsequent computed tomography and fluorodeoxyglycose positron emission tomography documented complete response to imatinib therapy. Our patient is now in sustained complete remission for 20 months with minimal toxicity. We conclude that sustained complete remission of metastatic DFSP with specific FISH abnormalities involving the PDGFB locus can be obtained with imatinib mesylate with minimal toxicity for the patient. PMID- 15746585 TI - Health care quality and disparities: lessons from the first national reports. PMID- 15746586 TI - Measurement challenges in developing the national healthcare quality report and the national healthcare disparities report. AB - OBJECTIVES: The objective of this study was to describe 2 measurement challenges faced in the development of the National Healthcare Quality Report (NHQR) and the National Healthcare Disparities Report (NHDR): the use of federal data on race and ethnicity and the selection of measures of socioeconomic status (SES). METHODS: Over 30 federal and nonfederal data systems were examined to identify measures of race, ethnicity, and SES and to evaluate the characteristics and relative quality of the data. RESULTS: The availability and quality of data on race, ethnicity, and SES vary by factors such as the type of data (population or establishment based-survey, administrative/claims data, or vital statistics), the source of information (self, proxy, other, or some combination), and the transition to new federal standards. No single measure of SES could be identified, so a mix of measures is presented, including income, education, and expected source of payment (ESOP). Income relative to federal poverty level was used as the preferred SES measure from person-based surveys. Selected analyses linking hospital discharge data to annual median household income from US census data were presented for data derived from administrative data systems. Educational attainment was the variable used for examining SES using data from the Vital Statistics System. CONCLUSIONS: The first NHQR and NHDR maximized the presentation of data by accommodating the variation among data systems while at the same time imposing some standardization in the coding and classification of data on race, ethnicity, and SES. PMID- 15746587 TI - Challenges in measuring nursing home and home health quality: lessons from the First National Healthcare Quality Report. AB - BACKGROUND: The availability of patient assessment data collected by all Medicare and Medicaid-certified nursing homes (NHs) (the Minimum Data Set [MDS]) and home health agencies (HHAs) (the Outcome and Assessment Information Set [OASIS]) provides an opportunity to measure quality of care in these settings. OBJECTIVE: The objective of this study was to examine methodologic issues encountered as these datasets are used to report the nation's health care in the National Healthcare Quality Report (NHQR) at national and state levels. FINDINGS: Although the reliability of most data elements from MDS and OASIS are considered acceptable in research studies, mixed evidence exists for the reliability and validity of the quality measures themselves. Detection bias can affect the quality measures, particularly for pain and pressure ulcers. Although risk adjustment is used for all measures, effectiveness varies among measures and methods. Additional quality measures such as patient satisfaction, quality of life, and structural measures would be desirable but will require additional data collection efforts. Although the NH measures represent most NH residents, the HHA measures only apply to Medicare and Medicaid patients served by Medicare certified agencies. Finally, the absence of clinical benchmarks limits the interpretation of the NHQR HHA and NH measures. CONCLUSIONS: Further developmental work is needed to address many of these issues to improve the usefulness of these quality measures in future NHQR reports. PMID- 15746588 TI - The national healthcare quality and disparities reports: an overview. AB - BACKGROUND: Congress directed the Agency for Healthcare Research and Quality (AHRQ) to lead an effort for the US Department of Health and Human Services (DHHS) to develop 2 annual reports: a National Healthcare Quality Report (NHQR) and a National Healthcare Disparities Report (NHDR). OBJECTIVES: This article lays out key concepts, definitions, statistical methods, and findings from these first ever national reports on quality and disparities. We also summarize some possible future directions for the reports. RESEARCH DESIGN: The NHQR and NHDR rely on secondary analysis of available data from over 40 established, national databases. The NHQR presents data at the national level, by sociodemographic characteristics, and at the state level. The NHDR presents data broken out by race/ethnicity and by socioeconomic status. MEASURES: The 2003 NHQR presented data on approximately 140 quality measures and the NHDR presented data on these same measures plus approximately 100 measures of access to care. RESULTS: The reports found that high healthcare quality is not a given and that disparities are pervasive throughout the US healthcare system. In addition, they found the quality and disparities issues are particularly apparent in preventive care, but that greater improvement is possible. CONCLUSIONS: As these reports evolve for the 2004 version and beyond, they will be a vital step in the effort to improve healthcare quality for all populations in the United States. PMID- 15746589 TI - Heart disease and prevention: race and age differences in heart disease prevention, treatment, and mortality. AB - OBJECTIVE: The objective of this study was to analyze race and age differences in the distribution of health promotion and cardiovascular screening tests, and the prevalence of serious heart disease and cardiovascular mortality in the United States. DATA SOURCES/STUDY POPULATION: Data are from 7 federal datasets represented in the first National Healthcare Quality Report and the National Healthcare Disparities Report, and include surveys, administrative and vital statistics data systems. The study analyzes blacks and whites. MEASURES: Counseling on diet and nutrition, exercise, and tobacco during an outpatient visit indicate the availability of health promotion services, and screening for high blood pressure and cholesterol represent preventive services. Hospitalizations for heart-related conditions and use of certain cardiac procedures identify serious heart disease. Deaths from coronary artery disease and stroke are the heart-related mortality measures. PRINCIPAL FINDINGS: Counseling and education services tend to occur more on outpatient visits by individuals aged 45 to 64 years than in younger age groups. Screening rates among individuals aged 45 to 64 years of approximately 90% for hypertension and 80% for high cholesterol suggest progress in early detection of cardiac risk factors. However, blacks aged 45 to 64 years are 5.6 times more likely than their white counterparts to be hospitalized for hypertension, approximately one third less likely to receive a cardiac procedure, and almost twice as likely to die of coronary heart disease. CONCLUSIONS: Although findings indicate few racial differences in health promotion services in ambulatory care or screening for cardiac risk factors, the prevalence of serious cardiovascular disease, use of cardiac procedures, and heart-related mortality suggest continuing racial disparities in heart disease. PMID- 15746590 TI - Assessing patient safety in the United States: challenges and opportunities. AB - BACKGROUND: In 1999, the US Congress mandated the Agency for Healthcare Research and Quality (AHRQ), Department of Health and Human Services (DHHS), to report annually to the nation about healthcare quality. One chapter in the National Healthcare Quality Report (NHQR) is focused on patient safety. OBJECTIVES: The objectives of this study were to describe the challenges in reporting the national status on patient safety for the first NHQR and discuss emerging opportunities to improve the comprehensiveness and reliability of future reporting. RESEARCH DESIGN: This study is a selective review of definitions, frameworks, data sources, measures, and emerging developments for assessing patient safety in the United States. RESULTS: Available data and measures for patient safety assessment in the nation are inadequate, especially for comparing regions and subpopulations and for trend analysis. However, many opportunities are emerging from the recently increased investments in patient safety research and many ongoing safety improvement efforts in the private sector and at the federal, state, and local government levels. CONCLUSION: There are many challenges in assessing national performance on patient safety today. Ongoing developments on multiple fronts will provide data and measures for more accurate and more comprehensive assessments of patient safety for future NHQRs. PMID- 15746591 TI - Racial, ethnic, and socioeconomic disparities in estimates of AHRQ patient safety indicators. AB - BACKGROUND: Patient safety events that result from the happenstance of mistakes and errors should not occur systematically across racial, ethnic, or socioeconomic subgroups. OBJECTIVE: To determine whether racial and ethnic differences in patient safety events disappear when income (a proxy for socioeconomic status) is taken into account. RESEARCH DESIGN: This study analyzes administrative data from community hospitals in 16 states with reliable race/ethnicity measures in the 2000 Healthcare Cost and Utilization Project of the Agency for Healthcare Research and Quality (AHRQ), using the publicly available AHRQ patient safety indicators (PSIs). RESULTS: Different indicators show different results for different racial/ethnic subgroups. Many events with higher rates for non-Hispanic blacks (compared with non-Hispanic whites) remain higher when income is taken into account, although such differences for Hispanics or Asian/Pacific Islanders (APIs) tend to disappear. Many events with lower rates for Hispanics and APIs remain lower than whites when income is taken into account, but for blacks, they disappear. DISCUSSION: The higher rates for minorities that reflect the way health care is delivered raise troubling questions about potential racial/ethnic bias and discrimination in the US health care system, problems with cultural sensitivity and effective communication, and access to high-quality health care providers. CONCLUSIONS: The AHRQ PSIs are a broad screen for potential safety events that point to needed improvement in the quality of care for specific populations. PMID- 15746592 TI - Children's Health Care in the First National Healthcare Quality Report and National Healthcare Disparities Report. AB - BACKGROUND: The first National Healthcare Quality Report (NHQR) and National Healthcare Disparities Report (NHDR) are landmark events for children's health care quality and are expected to stimulate local measurement, benchmarking, and quality improvement efforts. METHOD: The authors select findings from the NHQR and NHDR, focusing on topics reflecting a range of health care and health care settings affecting children. They highlight disparities by race/ethnicity, socioeconomic status, and insurance source. They critique the first NHQR and NHDR from a child health perspective. SELECT NHQR/DR FINDINGS: Quality-of-care issues in the effectiveness domain were identified for black infant mortality, low and very low birth weight rates, antibiotic use for the common cold, and childhood hospitalizations for asthma. Immunization rates have improved. Patient centeredness and timeliness results vary by race, ethnicity and income. The NHDR found that Hispanic and low-income children are most likely to be uninsured for part of the year. Groups of children most likely to have public coverage are American Indian/Alaska native, black, and Hispanic. CRITIQUE OF REPORTS: The structure and criteria used for the first reports limit their usefulness from a child health perspective. A basic problem is that the conceptualizations of health and health care that are driving national initiatives on quality are based largely on an adult chronic care model focused on conditions with high expenditures as treated in the mainstream health care delivery system. CONCLUSION: NHQR and NHDR provide essential information on children's health care quality. Future reports can be improved by including child-relevant perspectives in priority-setting and data-gathering efforts. PMID- 15746593 TI - NHQR/NHDR measures for women of reproductive age. AB - BACKGROUND: This article addresses measures of importance to women of reproductive age in the first National Healthcare Quality Report (NHQR) and National Healthcare Disparities Report (NHDR). METHODS: The authors review each of the 4 components of quality of care: effectiveness, safety, timeliness, and patient centeredness. The effectiveness component topics with relevance to women of childbearing age include breast cancer, cervical cancer, HIV, AIDS, mental health, and maternity care. The safety component includes 3 relevant measures of obstetric trauma. The quality aspects of timeliness of care and patient centeredness will be discussed in terms of women, although the NHQR and NHDR did not include them as a separate topic because the data were so limited regarding women. FINDINGS: There is a foundation of knowledge about many aspects of quality health care for women of reproductive age. However, gaps are evident in some measures, usually due to insufficient data. CONCLUSION: Further development of the measure set would benefit from additional process and outcome variables that can link screening, diagnosis, and treatment with health outcomes. Such linkages will expand our knowledge and capability to improve health outcomes for women of reproductive age. PMID- 15746594 TI - Variation in quality of men's health care by race/ethnicity and social class. AB - BACKGROUND: Until recently, minority and poor men have been characterized as "an invisible population," overlooked by public and private efforts to improve the health status of women, children, and the elderly. OBJECTIVE: This study compares the health care experiences of racial and ethnic minority men with that of white men, and low socioeconomic status with those of higher status. MEASURES/SUBJECTS: Quality-of-care measures in multiple clinical domains are evaluated. The authors use data from several databases, including the National Health Interview Survey, Medical Expenditure Panel Survey, and Health Care Cost and Utilization Project State Inpatient Database. The relative difference between each racial/ethnic and socioeconomic group and a fixed reference group is used to assess differences in use of services. Statistical significance is assessed using z tests. RESULTS: Hispanic men were much less likely to receive colorectal cancer screening (relative risk [RR] range, 0.61-0.69), cardiovascular risk factor screening and management (RR, 0.84-0.88), and vaccinations (RR, 0.47-0.94). Black and Asian men were significantly less likely to have received selected preventive services (adult immunization and colorectal cancer screening). The differences in end stage renal disease care that black and white men received were statistically significant (RR, 0.39-0.97), with black men consistently receiving worse care. For some measures of management of end-stage renal disease, Asian men received care that was similar to or better than that received by non-Hispanic whites. CONCLUSION: Minority men are at a markedly elevated risk for the receipt of poor health care quality. However, generalizations about "minority" men are likely to be misleading and incomplete. There is a considerable variation in the magnitude, direction, and significance of these risks. PMID- 15746595 TI - How the national healthcare quality and disparities reports can catalyze quality improvement. AB - BACKGROUND: The purpose of the National Reports on Healthcare Quality and Disparities is to enhance awareness of quality and health care disparities, track progress, understand variations, and catalyze improvements in health care. OBJECTIVES: The objective of this paper is to propose a model that will facilitate a user's progression from knowledge to action and to show how the reports, its data warehouse, associated products, and Agency for Healthcare Research and Quality resources are integrated and focused on a comprehensive campaign to improve health care quality. DESIGN: The design of the paper is to present a conceptual model and to show how implementation strategies for the reports fit the model. FINDINGS: The authors propose a quality improvement supply chain model to help elucidate the links of the process, corresponding developmental stages that potential users need to master and progress through, and "just-in-time" supply chain inputs at each of the corresponding stages, and populate the model with examples. CONCLUSION: The traditional ways of disseminating knowledge derived from science through reports and conferences are inadequate to the humbling need for vast improvements in the US health care system. Our model suggests the need for a wide variety of information, packaged in a diverse ways, and delivered just in time and on demand. It encourages the alignment of decision makers and researchers, along with information intermediaries and innovation brokers, to make the information production cycle more efficient and effective. Future iterations of the reports will improve relevance, meaning, and distribution of information to facilitate its uptake by potential users. PMID- 15746596 TI - Preparing the national healthcare disparities report: gaps in data for assessing racial, ethnic, and socioeconomic disparities in health care. AB - BACKGROUND: Efforts to quantify, monitor, understand, and reduce disparities in health care are critically dependent on the collection of high-quality data that support such analyses. In producing the first National Healthcare Disparities Report (NHDR), a number of gaps in data were encountered that limited the ability to assess racial, ethnic, and socioeconomic disparities in health care. OBJECTIVES: The objectives of this study were to identify and quantify gaps in data related to disparities in health care and discuss efforts to fill these gaps in future NHDRs. FINDINGS: : Data on specific racial, ethnic, and socioeconomic groups were often not collected or collected in formats that differed from federal standards. When collected, data were often insufficient to generate reliable estimates for specific racial, ethnic, and socioeconomic groups. These effects were magnified when attempting to assess disparities within many of the agency's priority populations such as women, children, the elderly, low-income populations, and rural residents. Future NHDRs begin to fill some of these gaps in data, but some gaps will likely persist and new gaps will likely arise as the availability of data for specific populations vary from year to year. CONCLUSIONS: Gaps in data limit the ability to address racial, ethnic, and socioeconomic disparities in health care. Although many federal efforts are underway to improve data collection, some groups and populations pose unique challenges for data collection that will be difficult to overcome. PMID- 15746597 TI - [Best wishes for 2005]. PMID- 15746598 TI - [Mycophenolate: 30 years later]. PMID- 15746599 TI - [Antibiotics and the risk of breast cancer]. PMID- 15746600 TI - [Cicatricial pemphigoid: treatment with mycophenolate mofetil]. AB - BACKGROUND: The severity of cicatricial pemphigoid (CP) varies. First-intent treatment of mild or moderate cases is dapsone. In life or sight-threatening cases, intravenous cyclophosphamide pulses are efficient but may have digestive side effects and imply repeated hospitalizations. Mycophenolate mofetil (MMF) is an oral and well tolerated immunosuppressant agent which has proved its efficacy in pemphigus and some bullous pemphigoid. In CP, encouraging case reports have been previously published. We report herein a retrospective study about 14 patients who have received MMF since 2000. PATIENTS AND METHODS: There were 5 men and 9 women, with a mean age of 69 years. MMF was introduced in 3 different clinical situations: immediately in relay to cyclophosphamide in 7 patients with severe CP (group I); in case of a mild-severe relapse at distance from with dranal of cyclophosphamide in 3 patients (group II); as first-intent immunosuppressant agent in 4 patients whose disease was not under control with high-dose dapsone, but not life - or sight-threatening (group III). In all these patients, the disease was invalidating and not controlled by dapsone +/- sulfasalazine, but did not threaten life or sight. The aim was to achieve satisfying control of the disease with an oral and well tolerated immunosuppressant agent, and to maintain good quality of life. The dose of MMF was 1.5 or 2 g per day. The criteria of MMF efficacy was the healing of previous lesions and the absence of new progressive lesions. RESULTS: MMF was efficient in obtaining or maintaining a good control of the disease in 10/14 patients, as long as the underlying treatment with dapsone (2 mg/kg/d) was maintained. In 7/10 cases, it was possible to decrease the dapsone dose in order to improve hematological tolerance. In the 3 other cases, a relapse occurred when the dose of dapsone was decreased. MMF was inefficient in controling the disease in 4/14 patients (29 p. 100). Clinical and biological tolerance of MMF was good in 13/14 patients. DISCUSSION: In this series, MMF was proposed to heterogenous patients, who presented at that time a mild-moderate disease and for whom we wanted in improve the quality of life. MMF seems to be an interesting drug, capable of obtaining or maintaining satisfactory control of the disease and permitting the decrease of dapsone doses in some mild-severe CP. However MMF must not replace cyclophosphamide in severe sight or life-threatening forms of CP. PMID- 15746601 TI - [Narrow band UVB phototherapy in the treatment of widespread lichen planus]. AB - INTRODUCTION: UVA phototherapy, acitretin and oral corticosteroids are currently the front-line treatment of disseminated cutaneous lichen planus. We studied the efficacy of narrow band UVB therapy in this indication. PATIENTS AND METHODS: We retrospectively studied the dossiers of patients suffering from disseminated cutaneous lichen planus, treated with narrow band phototherapy in the Phototherapy Unit of the University hospital in Montpellier, from May to November of the year 2001. Disseminated lichen planus was defined as lichen involving at least 20p. 100 of the skin surface. Twenty patients were included. UVB were applied thrice weekly using a Philips TL01 cubicle (311-313 nm). The protocol was that used for the treatment of psoriasis. We defined 4 types of response: complete response (disappearance of more than 90p. 100 of the lesions), partial response (disappearance of at least 50p. 100) poor response (improvement in 20 to 50p. 100) and failure (less than 20p. 100 reduction in the lesions). Assessment of relapses in the long term was made using a telephone survey among the patients treated or their physicians. RESULTS: Complete response was obtained in 11 out of the 20 patients (55p. 100) and partial response in 4 (20p. 100), corresponding to 75p. 100 of the responders. Response was obtained with a median delay of 3 months, ranging from 2 to 6 months, following a median of 30 sessions (12 to 50) and accumulated dose of UVB of 36 +/- 4.8 joules/cm2. The phototype, gender, age and duration of evolution before treatment did not influence the response. The relapse rate was and estimated 18p. 100 (2/11) 42 months after treatment had been stopped. DISCUSSION: In our opinion, these results underline the efficacy of narrow band UVB in the treatment of disseminated cutaneous lichen planus. They confirm those of earlier studies and are superimposable with those of oral UVA phototherapy. PMID- 15746602 TI - [Lymphedema of male external genitalia: a retrospective study of 33 cases]. AB - INTRODUCTION: The aim of this retrospective study was to describe the main characteristics and treatment of male external genitalia lymphedema. PATIENTS AND METHODS: From 1987 to 2003, all patients seen in a single hospital for lymphedema of male external genitalia were included. For each patient, the following characteristics were recorded: primary or secondary lymphedema, cause of secondary form, date of onset of lymphedema, associated lower limb lymphedema, clinical signs, and complications. In the primary forms, lower limb lymphoscintigraphy was performed. Specific surgery was proposed in all cases of symptomatic lymphedema (circumcision, scrotum and/or penile cutaneous excision). RESULTS: Thirty-three patients with lymphedema of external genitalia (17 primary, 16 secondary) were recruited. Two primary lymphedema were congenital, one isolated. Mean age +/- SD of the onset of the 15 other primary genital lymphedema was 23.4 +/- 17.5 years, always after the appearance of lower limb lymphedema. Sixteen men had secondary lymphedema (bladder, prostate, or rectum cancer, Hodgkin or non-Hodgkin lymphoma, aorto-bifemoral bypass grafting, biopsy or curretage of inguinal nodes). Secondary genitalia lymphedema was not associated with lower limb lymphedema in two cases and, in the others it occurred 66 +/- 122 months after (n=11), at the same time (n=2) or before lower limb lymphedema (n=1). Clinically, we noted genitalia heaviness (n=31), lower limb lymphedema (n=30), vaginal hydrocele (n=13), impaired miction due to prepucial swelling (n=10), leakage of lymphatic fluid (n=10). Lower limb lymphedema was complicated by at least one erysipelas (n=20), spreading to the external genitalia (n=4). In primary forms, lymphoscintigraphy showed ipsilateral hypoplasia of inguinal nodes in lower limb lymphedema (n=14) and/or external genitalia backflow (n=7). Surgical treatment was performed in 17 cases (11 primary, 6 secondary) with good results after 21 months' median follow up (1 month-10 years). Two patients died of cancer. One secondary lymphedema improved spontaneously and one disappeared after withdrawal of lower limb pneumatic compression. DISCUSSION: Lymphedema of external genitalia is responsible for discomfort and local complications. Surgical treatment is the main procedure of this disorder. PMID- 15746603 TI - [Neutrophilic dermatosis associated with propylthiouracil-induced p-ANCA (p antineutrophil cytoplasmic antibodies)]. AB - INTRODUCTION: We report on a patient who progressively developed polymorphic expressions of neutrophilic dermatosis (Sneddon-Wilkinson subcorneal pustulosis and pyoderma gangrenosum) associated with p-antineutrophil cytoplasmic antibodies (p-ANCA), while receiving propylthiouracil for hyperthyroidism. To our knowledge, such associations have never been published so far. CASE-REPORT: A 40 year-old woman was treated with propylthiouracil for Graves'disease. After 16 months of therapy, she noted flares of pustular lesions surrounded with erythematous halo mainly localized on the trunk. The lesions became chronic, and were not improved by potent topical corticosteroids. When first seen in our department in February 2003, the eruption was typical of Sneddon-Wilkinson subcorneal pustulosis. This diagnosis was confirmed by the histological examination of a skin biopsy of a pustule. One month later, she developed an inflammatory progressively ulcerative lesion on the right ankle, typical of pyoderma gangrenosum. The diagnosis was confirmed by the histological examination of a skin biopsy taken on the evolving border of the lesion and showed polynuclear neutrophilic infiltration without vasculitis. Direct immunofluorescence was negative. The presence of serum anti myeloperoxydase p-ANCA was known for this patient since October 2002. No IgA monoclonal gammapathy was revealed on extensive biological check-up. Systemic oral corticosteroid therapy (1 mg/kg/day) dramatically improved skin lesions with complete healing within 8 weeks. DISCUSSION: Propylthiouracil is well known to induce the occurrence of ANCA in 20 to 64p. 100 of patients treated for Graves'disease. The mechanisms involved are badly recognized so far. Cutaneous vasculitis, glomerulonephritis and polychondritis may be clinically associated with those antibodies. Rare observations of neutrophilic dermatosis, mostly Sweet's syndrome, have been described in patients with propylthiouracil-induced ANCA. One case-report described a 44 year-old woman who developed pyoderma gangrenosum associated with propylthiouracil-induced p-ANCA. These manifestations usually appear within 2 years, as our patient. The data in the literature, allows us to report the polymorphic expressions of neutrophilic dermatosis in this patient with p-ANCA which could be related to propylthiouracil. Such association of Sneddon-Wilkinson subcorneal pustulosis and pyoderma gangrenosum with p-ANCA has never been described in this endocrinologic context so far. Furthermore we propose that neutrophilic dermatosis should be inscribed in the list of side effects induced by propylthiouracil therapy. PMID- 15746604 TI - [Sweet's syndrome in a child]. AB - INTRODUCTION: Sweet's syndrome was described for the first time in 1964. It is usually described in adults and remains rare in children. We report a case in a 23 month-old infant. OBSERVATION: A 23 month-old boy presented with diffuse papular, edematous annular plaques surrounded by vesicles and bullas. Laboratory examinations revealed neutrophilic polynuclear hyperleukocytosis, anemia and an inflammatory syndrome. The myelogram was rich and the abdominal sonography normal. Oral corticosteroids (2 mg/kg/d) led to spectacular improvement. After 19 days' treatment, the boy developed cervical adenopathies and hepatomegaly. The second myelogram was normal. Evolution under corticosteroids was good. DISCUSSION: Sweet's syndrome is exceptional in infants. The frequent association with a malignant blood disease should prompt appropriate investigations and prolonged surveillance. Systemic corticosteroid therapy is the reference. PMID- 15746605 TI - [Mycosis fungoides presenting as annular erythema]. AB - INTRODUCTION: Mycosis fungoides is a lymphoma, the classical clinical form of which involves erythematosquamous lesions. However, it can present various atypical aspects: hyper pigmentation or hypo pigmentation, suggestive of pyoderma gangrenosum or ichtyosis. We report a case of mycosis fungoides, unusual in its presentation in the form of centrifugal annular erythema. OBSERVATION: A 78 year old man had developed a parapsoriasis in plaques for more than 20 years. In May 2002 he consulted because of the recent infiltration of one of the plaques, without concomitant pruritus. The clinical examination revealed 3 lesions of the popliteal groove of the right groin and the left cheek suggestive of centrifugal annular erythema. Histology, revealing Pautrier microabscesses, was compatible with the diagnosis of mycosis fungoides. Evolution was marked by the spontaneous regression of the plaque on the face and remission of the other two plaques after local treatment with chloromethin and topical corticosteroids. Nevertheless, new plaques appeared despite continued treatment, combined with PUVA therapy sessions. DISCUSSION: When searching the literature, we only found one other case of mycosis fungoides, the clinical aspect of which was a centrifugal annular erythema, but in which the histological examination confirmed the diagnosis of mycosis fungoides. Our case report is also unusual in the clinical regression of the lesion on the face, without treatment; this has only been reported in two cases. Mycosis fungoides can appear in various clinical forms. The centrifugal annular erythema form is rare, but this diagnosis should be evoked. PMID- 15746606 TI - [Recurrent Castleman's disease of the neck occurring in an infant]. AB - INTRODUCTION: Castleman's disease is a lympho-proliferative disease of unknown cause. This rare disease, usually localized, is benign and develops in the young adult. The predominant localization is mediastinal. OBSERVATION: A 15 month-old infant was referred for a left latero-cervical mass that had developed since the age of 3 months. Other than this, the clinical examination was normal. Sonography, x-ray and tomodensitometry confirmed the unique nature of the lesion and the absence of loco-regional invasion. Histology following surgical exeresis found a nodule with multiple pseudo-follicular structures. These mitosis-rich structures were arranged concentrically around a vessel and were bordered by small lymphocytes forming a crown. This histological aspect corresponded to Castleman's disease in its vascular hyaline form. Local recurrence was reported twice, 2 and 8 years later and was treated each time with surgical exeresis. Histological examination was always similar to the first. DISCUSSION: Castleman's disease has rarely been reported in infants. The cervical location, at whatever age, is far rarer than the mediastinal form. Supplementary examinations are not always of interest and basically provide information on the isolated or multicentric nature. Surgical exeresis was justified because of the diagnostic doubts and the risk of compression. Diagnosis is based on anatomopathology. Relapses have rarely been described in the literature and raise the question of the potential malignant transformation after several relapses. PMID- 15746607 TI - [Paraneoplastic pemphigus revealing dendritic cell sarcoma originating from Castleman's disease of the neck]. AB - INTRODUCTION: Paraneoplastic pemphigus is associated with Castleman's disease. We report a case of paraneoplastic pemphigus at the stage of the sarcomatous transformation of Castleman's disease, present for many years but without concomitant paraneoplastic pemphigus. The pemphigus was manifested by the most unusual, exclusive, involvement of the mucosa of the mouth and lung. OBSERVATION: A 32 year-old man suffering from extensive ulceration of the oral mucosa was hospitalized in December 2000 for alteration in his general status of health and acute respiratory failure. The search for intercellular anti-substance antibodies on rat spleen was positive, corresponding to anti-envoplakin IgG and leading to the diagnosis of paraneoplastic pemphigus. The thoracic x-ray and scan revealed a hilum tumor, the histological examination of which confirmed the diagnosis of Castleman's disease concomitant to sarcomatous transformation. Following surgical treatment, the respiratory failure worsened. The patient improved with systemic corticosteroids at the dose of 2 mg/kg/d and chemotherapy was initiated. The patient died suddenly within the context of acute respiratory failure, three months after surgery. DISCUSSION: This is a case of paraneoplastic pemphigus of unusual clinical and biological expression: exclusively mucosal involvement with obliterating bronchiolitis, explained by the isolated presence of antibodies recognizing envoplakin, without anti-desmoglein. The transformation of the Castleman tumor into a sarcoma may have unmasked intra-cellular antigens (plakins), initiating the specific immune reaction. PMID- 15746608 TI - [Application of genetic epidemiology to dissecting susceptibility to leprosy]. PMID- 15746609 TI - [Levomepromazine-induced peripheral edema]. PMID- 15746610 TI - [Xeroderma pigmentosum in Subsaharan Africa]. PMID- 15746611 TI - [Feasability and tolerance of adjuvant treatment of melanoma with immunotherapy according to Kirkwood protocol]. PMID- 15746612 TI - [Clinically amyotrophic dermatomyositis with fatal interstitial lung disease]. PMID- 15746613 TI - [Tumor of the umbilicus]. PMID- 15746614 TI - [Linear plantar erythematous-squamous plaque]. PMID- 15746615 TI - [Cutaneous amyloidosis]. PMID- 15746616 TI - [What imaging studies are necessary in limb lymphedema?]. PMID- 15746618 TI - [Laser and intense pulsed light for hair removal]. PMID- 15746619 TI - [Management of atopic dermatitis of the child. Short version]. PMID- 15746620 TI - [Mutations of tumor suppressor genes coding for folliculin in Birt Hogg Dube syndrome]. PMID- 15746621 TI - [Henri the hairy and his sister Antoinette]. PMID- 15746622 TI - [Buschke-Lowenstein tumor]. PMID- 15746623 TI - [Metamizole and pseudo-ephedrine induced acute generalized exanthematous pustulosis]. PMID- 15746631 TI - The Carbomedics aortic heart valve prosthesis: a review. AB - The Carbomedics bileaflet mechanical prosthesis was introduced in 1986 and until now more than 500,000 valves have been implanted. The aim of this study was to review the papers published on the Carbomedics aortic heart valve prosthesis. The Carbomedics prosthesis has a solid pyrolite carbon housing and flat leaflets of pyrolite carbon coated graphite that is impregnated with tungsten. The pyrolite carbon housing is reinforced by an outer stiffening ring composed of titanium which virtually eliminates the risk of leaflet escape. The design further enables valve rotation after implantation. The standard aortic valve prosthesis has the sewing cuff located at the outflow level of the valve cylinder. A high performance valve, the Carbomedics TopHat valve for supraannular implantation, is a standard aortic valve prosthesis where the sewing cuff has been transferred to the inflow level of the valve cylinder. A 2 size increase in valve size can be achieved by using the TopHat valve which is very important in patient-prosthesis mismatch. With the recommended international normalised ratio (INR) level for the Carbomedics aortic heart valve prosthesis the rate of embolic and bleeding events are low. Thrombosis of a Carbomedics aortic heart valve prosthesis is rarely seen and is the result of inappropriate anticoagulation without pannus formation. The incidence of prosthetic endocarditis is very low and this is also the case for noninfectious paravalvular leakage necessitating reoperation. Intrinsic dysfunction and/or structural failure has never been reported. The total absence of intrinsic dysfunction and structural failure of the Carbomedics aortic valve prosthesis is a great advantage. Further the complication rate is very low. The TopHat, the supraannular version of this prosthesis, is a perfect solution in patient-prosthesis mismatch. PMID- 15746632 TI - Intermittent aortic cross-clamping for coronary artery bypass grafting: a review of a safe, fast, simple, and successful technique. AB - Since the very beginning of coronary artery bypass grafting, the search for optimal myocardial protection has fascinated both clinicians and basic researchers. This retrospective review of a large patient cohort aims to display the advantages of one of the protective procedures, namely simple, intermittent aortic cross-clamping (IAC). Thus, this review aims to significantly contribute to daily bypass surgery. This review reports on coronary patients who were all operated on in international centers using IAC such that this review presents the state of the art on IAC. In addition, this review reports on the usage of IAC for more than 2 decades in the clinic of Dr. Bircks, Duesseldorf (DE) and the clinics of his former students. A meta-analysis of published data of international centers summarizes 7 837 operated patients with a total mortality of 123 (=1.6%). This excellent outcome compares well to the results of the Bircks'-related centers, where between 1978 and 2001, a total of 41 573 patients were revascularized with the help of IAC according to the original protocol. The total mortality was 778 (1.9%), with the lowest mortality rate (1.2%) in the largest center (Bad Oeynhausen, DE). According to the presented experience, IAC for coronary revascularization proves to be a highly effective method for myocardial protection; it has convincingly proven to be simple, safe and cost-efficient. PMID- 15746633 TI - Beneficial effect of aspirin on renal function in patients with renal insufficiency postcardiac surgery. AB - AIM: Renal function is one of the most important prognostic factors following cardiac surgery. Whether aspirin affects cardiopulmonary bypass related renal injury is investigated in this study. METHODS: Ninety-four patients with impaired renal function (creatinine = or >1.5 mg/dl) undergoing coronary artery bypass grafting (CABG) were categorized into 2 groups according to aspirin administration before surgery. Serum creatinine, urinary output and creatinine clearance along with other perioperative factors were compared between the 2 groups prior to surgery, 24 hours and 48 hours following cardiopulmonary bypass. RESULTS: Creatinine levels increased significantly in the second postoperative day only in the non-aspirin (control) group (3.7+/-1.6 vs 2.9+/-1.7 mg/dl, p=0.03). Aspirin (study) group had lower creatinine levels in day 1 (p=0.03) and day 2 (p=0.001). Furthermore, in the study group creatinine clearance was higher in day 1 (34.3+/-14.3 vs 30.9+/-13.1 ml/min, p=0.01) and in day 2 (32.6+/-13.8 vs 26.4+/-9.8 ml, p<0.0001). Creatinine levels at discharge were elevated compared to the preoperative levels in the control group (p=0.01). However, the study group had lower creatinine levels at discharge (2.6+/-1.4 vs 3.8+/-1.6 mg/dl, p<0.0001). Urinary output was higher in the study group in the first postoperative day compared to the control group (p=0.01). Postoperative bleeding was slightly increased in the study group compared to the control group (760+/ 230 ml vs 530+/-210 ml, p=0.01). CONCLUSIONS: Continuation of aspirin administration until the day of surgery may have a protective effect against renal injury resulting from cardiopulmonary bypass, with only a negligible increase in bleeding. Possible explanations for this effect are antiplatelet activity of aspirin during cardiopulmonary bypass causing inhibition of vasoconstrictive agents like thromboxane, and improvement of renal perfusion by reducing blood viscosity. PMID- 15746634 TI - Inclusion of lidocaine in cardioplegic solutions provides additional myocardial protection. AB - AIM: Lidocaine inhibits depolarization by blocking sodium and calcium influx and potassium release, abolishing the action potentials of cells in the Hiss-Purkinje system and myocit cells. As it can directly influence cardiac electric and mechanical activities, this study evaluated the efficacy of lidocaine in providing myocardial protection during normothermic blood cardioplegia. METHODS: Twenty-six dogs were randomly assigned to groups based on the cardioplegic induction solution they were to receive. Group I dogs (n=10) received a solution consisting of lidocaine (5 mg/kg), KCL (41.6 mEq/L) and 180 ml of normothermic blood. Group II dogs (n=10) received the same solution, except for the lidocaine and group III dogs (n=6) received only normothermic blood. In addition, 120 ml of normothermic blood was reinfused every 20 min. All dogs underwent cardiopulmonary bypass, 2 hours of global myocardial ischemia and 3 hours of reperfusion. Statistical differences were determined with the chi squared test, the two-way analysis of variance and Bonferroni's test. RESULTS: There were no deaths in group I. The survival rate in group II was 60%, and no dogs in group III survived (p=0.025). No difference in lactate liberation or left ventricular function (i.e., cardiac outflow and ejection fraction) was observed between groups. However, animals in group I demonstrated less enzymatic releases (troponin I, p=0.049 and CK, p=0.026) and less mitochondrial ultrastructural changes (p=0.022). CONCLUSIONS: Lidocaine offers myocardium additional protection against ischemia during cardiopulmonary bypass. PMID- 15746635 TI - Aortic valve replacement with the Cryolife-O'Brien stentless aortic bioprosthesis: five-year experience. AB - AIM: Improved hemodynamics with stentless bioprosthesis compared to stented valves have been well documented. It has been suggested that a simplified implant model, the Cryolife-O'Brien, offers less satisfactory outcomes compared with standard stentless models. This study was conducted to prospectively evaluate the midterm results after aortic valve replacement with the Cryolife-O'Brien stentless bioprosthesis. METHODS: In 1996, the prospective clinical trial using different stentless valves was initiated in our center. From September 1996 through August 2001, 132 consecutive patients with a mean age of 72.5 years underwent aortic valve replacement with the Cryolife-O'Brien porcine stentless bioprosthesis by the same surgeon. The predominant aortic valve lesion was stenosis in 110 cases and insufficiency in 22 cases. Patients have been followed up from 2 to 60 months, mean 28 months. Echocardiography was performed by the same echocardiographer preoperatively, intraoperatively, postoperatively at discharge, 2 to 6 months later and annually thereafter. RESULTS: Sixty-five percent of patients received a valve 25 mm in diameter or larger, 42% had concomitant coronary bypass grafting. The 30-day operative mortality rate was 6.8 %. Nine late deaths, none related to the valve, have occurred. Severe aortic insufficiency caused by oversizing led to early reoperation in 3 patients. The peak and mean systolic gradients decreased significantly during the first 12 months after implantation (p<0.001) and the effective valve areas increased significantly during this time interval (p<0.001). Eleven patients have aortic insufficiency, trivial in 7 and mild in 4. The actuarial survival at 5 years was 86+/-3%. The rate for freedom from endocarditis was 100% and for freedom from thromboembolic events 92%. CONCLUSIONS: The Cryolife-OBrien stentless bioprosthesis has superior hemodynamics and a low rate of valve-related complications thus representing a very good alternative to conventional stented bioprosthesis. The midterm results are encouraging but further follow-up is needed to determine the valve's durability. PMID- 15746636 TI - The effects of drainage with a Redon versus a conventional drain on postoperative pain and blood loss after valve replacements. AB - AIM: The aim of this study was to assess the influence of drainage with a Redon drain versus a conventional drain on postoperative pain and blood loss after valve replacements. METHODS: After approval by the local Ethics Committee and written informed consent, 30 patients, 20-60 years of age, scheduled for first elective valve replacement were included. After standardized anaesthetic regimens, cardiopulmonary bypass and coagulation therapy procedures and at the end of the operation, the patients were randomly assigned to 1 of 2 groups: (GI, n=15): drainage with 4 Redon drains; (GII, n=15): drainage with 2 conventional drains. Postoperative pain intensity at rest (VAS-R), during coughing and mobilization (VAS-M) in bed was independently evaluated using a visual analogue scale (VAS 100 mm) at 6 hourly intervals until 48 h after admission to the ICU (Ho). All patients received 2 g of paracetamol after obtaining the VAS score (8 g/24 h). No other analgesic agents were used. All patients were submitted to 2D echocardiography to verify the presence of pericardial effusion 24 h after surgery. Values are expressed as means. Pearson's chi squared and ANOVA (for repeated measurements) were used for statistical analysis. P<0.05 was considered significant. RESULTS: There was no statistically significant difference in the mean postoperative VAS-R and VAS-M, and in the mean postoperative total blood drainage 822.3 ml in GI, versus 704.3 ml in GII. Non pericardial effusion was found, and we did not see side-effects in any of the patients. CONCLUSIONS: We have shown that drainage with a Redon drains versus a conventional drain does not influence postoperative pain intensity and blood loss after valve replacements. PMID- 15746637 TI - Recombinant factor VIIa use in cardiac surgery--expanding the arsenal therapy for intractable bleeding? AB - A 72-year-old patient was admitted for mitral valve replacement because of infective endocarditis. Severe intractable bleeding in the early postoperative period was successfully treated with recombinant activated factor VII (rFVIIa). Thereafter, recovery was uneventful, and the patient was discharged on postoperative day 16. The current clinical aspects and experience of rFVIIa use in cardiac surgery are discussed. PMID- 15746638 TI - Solitary cardiac metastasis from a clear cell carcinoma of the kidney. A case report and review of the literature. AB - A case of solitary septal metastasis from a clear cell carcinoma of the kidney is reported in a 55-year-old man who 5 years before had undergone right nephrectomy and adrenalectomy. Since then, he had been successfully treated by means of chemotherapy, radiation therapy, or operated on, at almost yearly intervals, for secondary pancreatic, pulmonary and cerebral single metastases. Diagnosis was obtained by routine computed tomography. The septal mass was surgically removed and the patient was discharged on the 4th postoperative day. PMID- 15746639 TI - Anomalous left coronary artery from pulmonary artery: autogenous arterial tube for aortic implantation. AB - Many surgical procedures for the implantation of anomalous left coronary artery from pulmonary artery have been described. A dual coronary system offers most benefit and is most advantageous for the patient. Two autogenous flaps of the pulmonary trunk and ascending aorta, without mobilizing the coronary artery, were created for use in this procedure. This technique was successfully used in an 8 month-old patient with severe myocardial dysfunction and moderate mitral regurgitation. This procedure allows 2 coronary systems to be repaired for any anatomic change of the left coronary artery without the use of prosthetic material. There were no technical complications. During the postoperative course minimal inotropic support was used. The magnetic resonance study, 11 months postoperative, showed wood flow of the left coronary artery and mild mitral regurgitation. The operative technique was simple and its execution easy. The follow-up in the intermediate and late period showed the growth of this endothelized tube. PMID- 15746640 TI - Patent foramen ovale after lobectomy. A contraindication to completion pneumonectomy? AB - Patency of the foramen ovale (FO) is a very rare complication after lobectomy. Completion pneumonectomy after FO reopening has never been described before. In the reported case, a patent FO was diagnosed in a 52-year old man 9 months after a left upper sleeve lobectomy for a squamous cell carcinoma pT2N1. At the same time, bronchoscopy showed an endobronchial recurrence in the left main bronchus. The foramen was closed percutaneously by a 35 mm Amplatzer prosthesis. Dyspnea improved dramatically in the following 4 weeks and no more oxygen therapy was required. After complete restaging, a completion pneumonectomy was performed without any postoperative complication. This case report suggests that pneumonectomy in such delicate patients is feasible. PMID- 15746641 TI - Synchronous independent bifocal orthotopic thymomas. A case report. AB - A case of synchronous multiple thymomas is reported. The patient was an 81-year old woman with 2 separate thymomas in the anterior mediastinum. A histological study revealed that both thymomas consisted of dense lymphocyte infiltration in the stroma and a small number of polygonal epithelial tumor cells with clear nuclei and distinct nucleoli. Immunohistochemical staining using the antibodies to 2 antigens, bcl-2, and MIB-1 showed identical staining patterns. In spite of these findings, we considered their origin to be synchronous multicentric development rather than intra-thymic metastasis, based on the non-invasive nature of these tumors. PMID- 15746642 TI - Severe calcification of gastroepiploic artery in a dialysis patient. PMID- 15746643 TI - Common femoral artery transection by blunt trauma. PMID- 15746644 TI - Statins and abdominal aortic aneurysms. PMID- 15746645 TI - Double unilateral thoracotomy for malignant pleural mesothelioma. PMID- 15746646 TI - Peripheral tumor emboli to both lower extremity arteries after pneumonectomy for extensive pulmonary cancer. PMID- 15746647 TI - Linking thrombophilia and idiopathic intracranial hypertension. PMID- 15746648 TI - Searching for an efficient institutional review board review model: Interrelationship of trainee-investigators, funding, and initial approval. AB - In this study we sought to identify variables associated with institutional review board (IRB) decisions to develop an efficient "pre-IRB" review model. We explored several variables, including relationships among the identification of trainees as investigators, external sources of funding, and initial approval. The sample consisted of all new submissions reviewed by the 2 medical IRBs at the University of Miami (UM) during a 1-year period. Trainees included students, residents, and fellows. At least 1 trainee had to be identified for a proposal to be considered a trainee submission. The medical-science committees (MSCs) were similar with regard to the numbers of new submissions they reviewed during convened meetings (MSC-A 242, MSC-B 241) and the percentages of proposals that were initially approved (MSC-A 52.9%, MSC-B 53.1%). Approved submissions were defined as those initially approved or conditionally approved pending minor modifications. We noted a robust statistical difference between the percentages of trainee submissions initially approved (39.9%) and submissions that did not identify a trainee (59.4%) ( P <.0001). Of the proposals that were initially not approved (tabled [deferred] or rejected [not approved]), 28.9% of those including a trainee were rejected, compared with 11.0% without a trainee ( P <.001). Proposals in which the source of funding was identified were more likely to be approved (64.2%) than were those in which it was not (30.8%) ( P <.0001). Funding also seemed to influence the trainee and initial-approval interaction. Our results show that new submissions that identified trainee investigators were more likely to be deferred or not approved than those that did not. Nonapproved proposals that identified a trainee were 3.8 times more likely to be initially rejected than those that did not. A prereview model could target those submissions that list a trainee, lack funding, or both. PMID- 15746649 TI - Idiopathic intracranial hypertension, polycystic-ovary syndrome, and thrombophilia. AB - We studied thrombophilia, hypofibrinolysis, and polycystic-ovary syndrome (PCOS) in 65 women consecutively referred because of idiopathic intracranial hypertension (IIH) as a means of better understanding the origin of IIH, with the ultimate goal of developing novel medical therapies for IIH. Our hypothesis: IIH results in part from inadequate drainage of cerebrospinal fluid (CSF) resulting from thrombotic obstruction to CSF resorption-outflow, favored by thrombophilia hypofibrinolysis. We conducted the polymerase chain reaction (PCR) and assessed serologic coagulation measures in 65 women (64 of them white) with IIH, PCR in 102 healthy white female controls (72 children, 30 age-matched adults), and serologic measures in the 30 adults. Of the 65 patients, 37 (57%) were found to have PCOS; 16 (43%) were obese (BMI > or = 30 to < 40), and 19 (51%) were extremely obese (BMI > or = 40). Of the 65 women with IIH, 25 (38%) were homozygous for the thrombophilic C677T MTHFR mutation, compared with 14% of controls (14/102) ( P = .0002). Thrombophilic high concentrations of factor VIII (>150%) were present in 9 of 65 (14%) IIH cases, compared with 0 of 30 controls (0%) (Fisher's p [p f ] = .053). An increased concentration of lipoprotein A (> or = 35 mg/dL), associated with hypofibrinolysis, was present in 19 of 65 IIH cases (29%), compared with 3 of 30 controls (10%) (p f = .039). IIH occurred in 18 of 65 IIH patients taking estrogen-progestin contraceptives (28%), in 6 patients taking hormone-replacement therapy (9%), and in 5 pregnant subjects (8%). We speculate that PCOS, associated with obesity and extreme obesity, is a treatable promoter of IIH. We also speculate that if thrombophilia hypofibrinolysis and subsequent thrombosis are associated with reduced CSF resorption in the arachnoid villi of the brain, thrombophilia and hypofibrinolysis-often exacerbated by thrombophilic exogenous estrogens, pregnancy, or the paradoxical hyperestrogenemia of PCOS-are treatable promoters of IIH. PMID- 15746650 TI - Association between reduced low density lipoprotein oxidation and inhibition of monocyte chemoattractant protein-1 production in statin-treated subjects. AB - Monocyte chemoattractant protein-1 (MCP-1) is essential in atherogenesis. Oxidized lipids regulate MCP-1 expression and release from mononuclear cells. In this study we investigated (1) whether statin therapy reduces lipopolysaccharide (LPS)-stimulated MCP-1 production in human whole-blood samples and (2) the relationships between in vitro low-density lipoprotein (LDL) oxidation and MCP-1 production. Fasting blood samples were obtained from 55 healthy nonsmoking adults with moderate hypercholesterolemia who were participating in a randomized double blind 8-week trial comparing the effects of statin therapy with those of placebo on cytokine production. Samples were analyzed for resistance to copper-mediated LDL oxidation (lag time in minutes), as well as MCP-1- and interleukin-8 (IL-8) stimulated production. Statin therapy reduced MCP-1 production (mean +/- SD) -161 +/- 399 pg/mL/mm 3 white cells) compared with 267 +/- 985 pg/mL/mm 3 in the placebo group, but changes were not different between active and placebo groups ( P = .13). Statin therapy also increased lag times (median [interquartile range]; 20.5 [7.0-51.2] minutes vs -17.0 [-5.3-16.5] minutes; P = .067 for group difference). Inhibition of MCP-1 production correlated with prolongation of lag time ( r = .46, P = .0056) in statin-treated subjects. Statin therapy reduced MCP 1 production in the whole blood of human subjects and these changes were correlated with improvement in LDL oxidative resistance. PMID- 15746651 TI - Evaluation of interferon-gamma, interferon-gamma-inducing cytokines, and interferon-gamma-inducible chemokines in tuberculous pleural effusions. AB - Tuberculous and malignant pleural effusions are representative of lymphocytic pleural effusions. In tuberculous pleurisy, especially, T-helper type 1 (Th1) cytokines are dominant, containing, for example, high concentrations of interferon (IFN)-gamma. We focused on cytokines that induce expression of IFN gamma and Th1 cell-specific CXC chemokines induced by IFN-gamma. We also evaluated the diagnostic utility of these markers in tuberculous pleural effusions. Forty-three patients with pleural effusions (11 with tuberculous pleuritis, 32 with malignant pleuritis) were studied. We measured the pleural concentrations of IFN-gamma, IFN-gamma-inducing cytokines (interleukin IL-12 and IL-18), and IFN-gamma-inducible chemokines (interferon-gamma-inducible protein of 10-kD [IP-10], monokine induced by interferon-gamma [Mig], and interferon inducible T-cell alpha chemoattractant [I-TAC]). Our results demonstrate that the concentrations of IFN-gamma, IFN-gamma-inducing cytokines, and IFN-gamma inducible chemokines were all higher in tuberculous pleural effusions than in malignant pleural effusions. Also, IFN-gamma was significantly correlated with IL 12, Mig, and I-TAC. Moreover, receiver-operator-characteristic (ROC) analysis demonstrated that IFN-gamma produced a greater area under the ROC curve than any other factor. We conclude that the concentrations of IFN-gamma, cytokines that induce expression of IFN-gamma, and chemokines induced by IFN-gamma in tuberculous pleural effusion were all increased. The Th1 chemokines we examined, especially IP-10, are comparable to IFN-gamma as diagnostic markers of tuberculous and malignant pleural effusions, although IFN-gamma is the most valuable. PMID- 15746652 TI - Usefulness of brilliant cresyl blue staining as an auxiliary method of screening for alpha-thalassemia. AB - Hemoglobin (Hb) electrophoresis is widely used in thalassemia screening. Most Hb variants express a specific abnormal band on the cellulose acetate membrane. The technique is useful in the diagnosis of the type of thalassemia but is not sensitive enough to detect alpha-thalassemia minor because the quantity of the HbH is too small to be expressed on the supporting medium. We used simple staining of blood smears rather than the sophisticated molecular method to detect HbH inclusions. To evaluate the effectiveness of this method, we used brilliant cresyl blue (BCB) staining of red blood cells in 509 patients with microcytosis and erythrocytosis caused by various conditions. The results indicate that BCB staining is useful in the identification of subjects who possess alpha thalassemia traits. Coexisting conditions such as beta-thalassemia and iron deficiency anemia did not affect the detection of the HbH inclusions with the use of BCB staining. We conclude that BCB staining is helpful and reliable as an auxiliary method of detecting HbH inclusions in the diagnosis of alpha thalassemia traits, especially in places where medical resources are limited. PMID- 15746653 TI - Effects of hepatitis B virus X protein on human telomerase reverse transcriptase expression and activity in hepatoma cells. AB - In subjects with hepatitis B, carcinogenesis has been associated with the hepatitis B virus (HBV) X protein (HBX) and human telomerase reverse transcriptase (hTERT). In the experiments reported here, we used immunohistochemical methods to study the expression of hTERT and HBV antigens (HBsAg, HBcAg and HBxAg) in 34 cases of HCC and corresponding paratumor tissues, 30 cases of liver cirrhosis, and 6 normal livers. To examine the effect of HBX on hTERT expression and activity in hepatoma cells, we transiently and stably transfected the pCMV-X plasmid cloned HBx gene into H7402 hepatoma cells, then measured the expression of c-Myc and hTERT in these cells with the use of Western blot analysis. Telomerase activity was detected with the use of the telomerase repeat amplification protocol (TRAP) in transiently and stably transfected cells. We found that hTERT expression was 67.6%, 73.5%, and 100% in tumor, paratumor, and cirrhosis samples, respectively, but found no hTERT positivity in samples of normal liver. HBsAg, HBcAg, and HBxAg were expressed in 58.8%, 26.5%, and 76.5% of tumor tissues, respectively; in 64.7%, 41.2%, and 85.3% of the corresponding paratumor tissues; and in 76.7%, 66.7%, and 100% of cirrhotic tissues. The chi 2 test revealed no significant difference between the expression of hTERT and HBxAg in these tissues. Western-blot analysis revealed that expression of c-Myc and hTERT in the transiently transfected cells was much greater than that in the control cells. We elicited a similar result when we used the TRAP method to measure telomerase activity. Our data collectively demonstrate that HBX up regulates the expression and activity of hTERT in hepatoma cells, suggesting that hTERT is associated with tumor development. PMID- 15746654 TI - Regulation of alternative splicing of caspase-2 through an intracellular signaling pathway in response to pro-apoptotic stimuli. AB - Alternative splicing is an important mechanism in the generation of functionally distinct products from the same gene. Some apoptosis-regulating genes also undergo alternative splicing, generating splice variants that antagonzie normal transcripts on apoptosis. For example, caspase-2 is alternatively spliced, leading to exon 9-lacking caspase-2L (proapoptotic) and exon 9-containing caspase 2S (antiapoptotic) transcripts. Serine-arginine splicing factor proteins (SR proteins) are highly conserved and required for constitutive and alternative messenger RNA (mRNA) splicing. Their activity is regulated by reversible phosphorylation on serine residue. During apoptosis, many functional molecules undergo posttranslational modification, including phosphorylation, dephosphorylation, and caspase cleavage. In this study, we investigated the effect of proapoptotic stimuli on alternative splicing of caspase-2 mRNA in U937 cells. U937 cells were simulated with etoposide, staurosporine, pacritaxel, or cyclohexamide. We analzyed the alternative splicing of caspase-2 mRNA using reverse transcription-polymerase chain reaction. Etoposide, staurosporine, pacritaxel, and cyclohexamide treatment promoted exon-9 inclusion, increasing the ratio of caspase-2S to caspase-2L in a time-dependent manner. Pretreatment with calyculin A, an inhibitor of protein phosphatase-1, blocked etoposide-induced alternative splicing of caspase-2 mRNA. Furthermore, pretreatment of U937 cells with fumonisin B1, an inhibitor of ceramide synthase, also blocked alternative splicing of caspase-2 mRNA. These data demonstrate that endogenous ceramide generation and subsequent phosphatase activation during apoptosis are key steps in the alternative splicing of caspase-2 mRNA and further suggest a link between the signal-transduction pathway and alternative splicing. PMID- 15746655 TI - Fetal RhD typing with free DNA in maternal plasma. PMID- 15746656 TI - Fetal RhD genotyping by maternal serum analysis: a two-year experience. AB - OBJECTIVE: The purpose of this study was to determine the accuracy of the none invasive prenatal determination of polymerase chain reaction (PCR)-based fetal RhD genotyping. STUDY DESIGN: A prospective case series was undertaken on all RhD negative pregnant women presenting for genetic counseling in our prenatal diagnosis center from January 2001 until December 2002. Results were compared with serologic RhD typing of the newborns. RESULTS: Among the 285 pregnant women who participated in the study, fetal RhD status could be determined for 283 patients. In 2 cases, the RhD-negative phenotype of the mother was not the result of a complete RHD gene deletion, and therefore, the status of the fetus could not be determined. Neither false-negative nor false-positive results were observed. CONCLUSION: The present report demonstrates that a reliable fetal RHD genotype determination can be achieved with 100% accuracy. It is therefore possible to consider that such an assay could be systematically proposed to all RhD-negative pregnant women in order to more effectively utilize RhD prophylaxis. PMID- 15746657 TI - Prenatally diagnosed Down syndrome: mothers who continued their pregnancies evaluate their health care providers. AB - OBJECTIVE: This study was undertaken to ask mothers who had children with Down syndrome after receiving a prenatal diagnosis: How was the process and what, if anything, could be improved? STUDY DESIGN: An 11-page survey was mailed to 2945 persons on the membership lists of 5 Down syndrome parent organizations. The survey gathered both quantitative and qualitative data from yes/no questions, open-ended questions, and a series of statements asking the mothers to rate their level of agreement on a 1-to-7 Likert scale. Qualitative data were analyzed using the Constant Comparative Method of Qualitative Analysis, and quantitative data were summarized using linear regressions, mixed stepwise multiple regressions, and grouped means, 1-way analysis of variance analyses. RESULTS: Of 1126 surveys received, 141 (12.5%) were from mothers who had received a prenatal diagnosis. Though satisfied with the care that they had received, the majority of respondents expressed frustration with the process. The most common suggestions were that the diagnosis be conveyed in person, that up-to-date printed materials on Down syndrome (DS) be provided, and that mothers be referred to local DS support groups. CONCLUSION: Receiving a prenatal diagnosis of DS need not be a negative experience. By implementing suggestions proposed herein by the mothers, health care providers can even make the situation a positive one. PMID- 15746658 TI - A short cervix in women with preterm labor and intact membranes: a risk factor for microbial invasion of the amniotic cavity. AB - OBJECTIVE: The purpose of this study was to determine whether there was a relationship between sonographic cervical length and the presence of culture proven microbial invasion of the amniotic cavity in women with preterm labor and intact membranes. STUDY DESIGN: Ultrasonography and amniocentesis were performed in 401 patients admitted with preterm labor (22-35 weeks) and cervical dilatation of < or = 3 cm, as assessed by digital examination. Cervical length was determined by transvaginal ultrasound at admission. Outcome variables were the presence of microbial invasion of the amniotic cavity (defined as a positive amniotic fluid culture) and the occurrence of preterm delivery before 35 weeks. Contingency tables, chi2 test, receiver-operator characteristic (ROC) curves, and logistic regression were used for statistical analysis. RESULTS: The prevalence of microbial invasion of the amniotic cavity was 7% (28/401). Spontaneous preterm delivery (< or = 35 weeks) occurred in 21.4% (82/384) of patients. ROC curve analysis showed a significant relationship between the frequency of microbial invasion of the amniotic cavity and the length of the uterine cervix (area under the curve: 0.77; P < .005). Patients with a cervical length < 15 mm had a higher rate of a positive amniotic fluid culture than patients with a cervical length > or = 15 mm (26.3% [15/57] vs. 3.8% [13/344], respectively; P < .05). Moreover, patients with a short cervix (defined as < 15 mm) were more likely to deliver spontaneously before 35 weeks, 32 weeks, within 7 days, and within 48 hours of admission ( P < .05 for all comparisons). Forty percent of patients (161/401) had a cervical length > or = 30 mm. These patients had a very low risk of microbial invasion of the amniotic cavity (1.9% [3/161]), spontaneous delivery < or = 35 weeks (4.5% [7/154]), < or = 32 weeks (2.6% [2/76]), within 7 days (1.9% [3/154]), and within 48 hours (0% [0/154]) of admission. CONCLUSION: Endovaginal ultrasonographic examination of the uterine cervix in women with preterm labor identifies patients at increased risk for intrauterine infection. PMID- 15746659 TI - Controversies and uncertainties: abdominal versus vaginal surgery for pelvic organ prolapse. AB - Reconstructive pelvic surgery is a common phenomenon in American women. The efficacy and durability of current procedures are often extrapolated from clinical case series and may be fraught with bias. Although the route of reconstructive pelvic surgery is debated with little evidence to support expert opinion, several recent clinical trials have provided a modest amount of data that can assist surgeons in counseling women who are facing prolapse repairs. An individual woman's tolerance for certain symptoms and risks can be matched with the scientifically based evidence for certain procedures. It is clear that the route of surgery must vary with individual surgeons and individual patients. The challenge is to test current surgical habits using modern clinical trials to obtain the necessary information to optimize each and every woman's restorative surgery. PMID- 15746660 TI - A systematic review of the association between factor V Leiden or prothrombin gene variant and intrauterine growth restriction. AB - OBJECTIVE: The purpose of this study was to conduct a systematic review of the literature of studies that examined the association between factor V Leiden and/or prothrombin gene variant and intrauterine growth restriction. STUDY DESIGN: This systematic review of studies assesses the association between factor V Leiden and/or prothrombin gene variant and intrauterine growth restriction. RESULTS: Ten case-control studies fulfilled the selection criteria for inclusion in the meta-analysis. There was a significant association between factor V Leiden and intrauterine growth restriction (odds ratio, 2.7; 95% CI, 1.3-5.5) and prothrombin gene variant and intrauterine growth restriction (odds ratio, 2.5; 95% CI, 1.3-5.0). Five cohort studies were identified in the systematic review; 3 studies were prospective (2 full publications), and 2 studies were retrospective (1 full publication). Combining the 2 full publication prospective studies yields a summary relative risk of 0.99 (95% CI, 0.5-1.9). CONCLUSION: This meta-analysis of case-control studies suggests that the factor V Leiden and prothrombin gene variant both confer an increased risk of giving birth to an intrauterine growth restricted infant, although this may be driven by small, poor-quality studies that demonstrated extreme associations. Large well-conducted prospective cohort studies are required to determine definitively whether an association between thrombophilia and intrauterine growth restriction is present. PMID- 15746662 TI - Proteomic biomarkers that predict the clinical success of rescue cerclage. AB - OBJECTIVE: The origin of incompetent cervix is multifactorial, and the success of rescue cerclage is unpredictable. We tested amniotic fluid from women who were preparing to undergo rescue cerclage for proteomic biomarkers and correlated their presence with clinical outcome. STUDY DESIGN: Amniocentesis was performed to facilitate rescue cerclage in 37 consecutive women with painless dilation (> 2 cm) and no detectable uterine activity for 4 hours (range, 1-24 hours) before cerclage. Thirty-nine consecutive women with a sonographically normal pregnancy and cervix who underwent amniocentesis for chromosomal testing during the same study interval at the same clinical site provided the control samples. A proteomic fingerprint was generated with the discarded sample and the Mass Restricted score (MR score) for inflammation calculated. Peaks corresponding to free hemoglobin chains were sought as evidence of decidual hemorrhage or intra amniotic bleeding. RESULTS: Amniocentesis was performed at 23.5 weeks in cerclage (mean dilation, 4 cm) versus 19.5 weeks in control subjects. Cerclage subjects were delivered at 28.8 weeks; control subjects were delivered at 39.2 weeks. Thirty-two of 37 of cerclage subjects (86%) were delivered prematurely. Ten of 37 of cerclage subjects (27%), but no control subject, had a MR score that was indicative of inflammation (P < .001). Hemoglobin peaks were present in 12 of 37 of cerclage subject (32%), but no control subjects. Among cerclage subjects, those with a MR score of 3 to 4 were delivered earlier than those with a MR score of 0 to 2 (P < .001). Women with a MR score of 3 to 4 had a shorter latency period (days from amniocentesis to delivery; 3 days) and a shorter percentage of prolongation (1.8%) than women with a MR score of 0 to 2 (35 days; P < .05; 17.9%; P < .05). Women with hemoglobin had a shorter latency period (6 days) and a shorter percentage of prolongation (3.8%) than women without hemoglobin (38 days; P < .05; 21.8%; P < .05). Hemoglobin was present in 7 of 10 of the cerclage subjects (70%) with a MR score of 3 to 4. Women with both a MR score of 3 to 4 and hemoglobin had the shortest intervals to delivery. CONCLUSION: These findings illustrate 2 pathologic mechanisms that are associated with preterm delivery are also associated with incompetent cervix. Either an intrauterine inflammatory response or decidual hemorrhage predates surgery in one half the women whose condition requires rescue cerclage. The activation of either mechanism predicts cerclage failure. PMID- 15746663 TI - Dexamethasone prevents long-lasting learning impairment following a combination of lipopolysaccharide and hypoxia-ischemia in neonatal rats. AB - OBJECTIVE: There are no established therapies for preventing or rescuing perinatal infection or inflammation-induced perinatal brain damage. We administered dexamethasone (DEX), a synthetic corticosteroid anti-inflammatory drug, to neonatal rats in a model of such damage induced by a combination of lipopolysaccharide (LPS) and hypoxia-ischemia (HI), which produces characteristic histologic and behavioral abnormalities. STUDY DESIGN: Four hours after the injection of LPS (1 mg/kg, i.p.), 7-day-old Wistar rat pups were subjected to unilateral HI for 1 hour according to Levine's procedure. Injections of 0.5 mg/kg of dexamethasone (DEX-treated group, n = 15) or saline (saline-treated group, n = 15) were given 4 hours before HI. A sham-operated control group received neither LPS nor HI (n = 15). We chose rats of this age because their stage of brain maturation is similar to the human neonate. Over the 7 to 16 weeks after treatment, a choice reaction time (CRT) task was used for assessment of attention processes in each group, an 8-arm radial maze task was used to test short-term memory, and a water maze task was used to test long-term memory. In the CRT task, the reward food was released when the tested animal correctly pressed a lever on the side of an illuminating lamp. The correct and incorrect lever pressings were counted. In the 8-arm radial maze task, rats were allowed to move freely, seeking a reward of food placed at the end of 1 arm. An error was defined as the choice of an arm that had already been visited. In the water maze, rats had to swim to seek a concealed platform as aversive escape motivation. At 19 weeks, the rats were euthanized, the brain was removed, sectioned coronally, and the volume of each part was measured. RESULTS: The striatum, cortex, and hippocampus showed reductions in volume in the saline-treated group (42.7%, 49.2%, and 34.9% decreases compared with the sham-operated controls, respectively), but this was not observed in the DEX-treated group. All learning and memory processes were impaired with the combination of LPS and HI treatment, but these deficits were almost completely prevented by DEX treatment. CONCLUSION: Dexamethasone is a promising candidate for prevention of infection and inflammation-induced perinatal brain damage. The impact of dexamethasone identifies potential therapeutic pathways once the mechanism of dexamethasone's protection is determined. PMID- 15746664 TI - No phenotype associated with established lipopolysaccharide model for cerebral palsy. AB - OBJECTIVE: Cerebral palsy (CP) is associated with childhood spasticity, seizures, and paralysis. Oligodendrocyte damage resulting in periventicular leukomalacia (PVL) in the developing brain has been implicated. Animal models of CP have used prenatal hypoxia and infection with histopathology of PVL as the end point. To evaluate whether this histologic end point is associated with a CP phenotype, we reproduced a lipopolysaccharide (LPS) model for PVL, 1 and evaluated developmental, behavioral, and motor outcomes. STUDY DESIGN: On gestational day 15, Fischer 344 rats were intracervically injected with .1 mg/kg LPS (n = 5) or saline (n = 4). After delivery, evaluation for developmental milestones was performed on days 1 to 21 (LPS = 45; control = 30 pups). Males were also tested at 2.5 months using open-field, rotarod, and anxiety tests. On day 21, 2 pups/litter were perfused for immunohistochemistry, and stained with 2 oligodendrocyte antibodies: 2', d'-cyclic nucleotide phosphodiesterase (CNP), and myelin proteolipid protein (PLP) with relative densities of staining assessed using NIH Image software. Statistical analysis included Mann-Whitney U and analysis of variance (ANOVA). RESULTS: LPS pups demonstrated decreased CNP (P = .04) and PLP (P = .06) staining, replicating the model. There was no difference seen in neonatal weight, righting, negative geotaxis, cliff aversion, rooting, forelimb grasp, audio startle, air righting, eye opening, and activity. Surprisingly, LPS-exposed neonatal rats mastered forelimb placement (P < .01) and surface righting (P = .02) earlier than control rats. There were no differences between adult groups in open field distance traveled (P = .8), open-field locomotion time (P = .6), rotarod (P = .6), or anxiety (P = .7). CONCLUSION: Histologic evidence of white matter damage can be replicated using an LPS model for intrauterine inflammation. Significant phenotypic differences consistent with the motor and cognitive damage sequelae of such lesions (ie, CP) were not demonstrated. When evaluating animal models, it is important to assess not only biochemical markers for human disease, but also clinically relevant phenotypes. PMID- 15746666 TI - Absence of association of inherited thrombophilia with unexplained third trimester intrauterine fetal death. AB - OBJECTIVE: The purpose of this study was to investigate the alleged association between thrombophilia and unexplained third-trimester stillbirth. STUDY DESIGN: Case subjects were 37 women with a history of a third-trimester unexplained stillbirth. Control subjects were 46 volunteers, group-matched for ethnic origin, with no history of stillbirth, recurrent fetal loss, or thromboembolism. The pathology report of 34/37 placentas of case subjects was reviewed. RESULTS: The prevalence of at least 1 inherited thrombophilia among case subjects was 37.8% compared with 41.3% among control subjects. (OR = 0.87; 95%CI, 0.32-2.29). There was no significant difference between the groups with respect to the prevalence of any single inherited thrombophilia. There was, however, a significantly higher prevalence of antiphospholipid antibodies among case subjects compared with control subjects: 47.2% vs 8.7%, respectively (OR = 9.4; 95%CI, 2.5-42.3). No significant difference was noted in the prevalence of thrombopilia among subjects with or without placental infarcts. CONCLUSION: We did not find an association between unexplained third-trimester intrauterine fetal death and inherited thrombophilia; however, we did find such an association with antiphospholipid antibodies. PMID- 15746665 TI - Urinary angiogenic factors cluster hypertensive disorders and identify women with severe preeclampsia. AB - OBJECTIVE: Serum levels of soluble fms-like tyrosine kinase 1 (sFlt-1), vascular endothelial growth factor (VEGF), and placental growth factor (PlGF) are altered in women with clinical preeclampsia. We sought to identify whether similar alterations in urinary levels of these proteins cluster hypertensive disorders in pregnancy, and identify women with severe preeclampsia (sPE). STUDY DESIGN: Free urinary levels of sFlt-1, VEGF, and PlGF were measured by immunoassay in 68 women enrolled prospectively in the following groups: nonpregnant reproductive age (NP CTR n = 14), healthy pregnant control (P-CTR n = 16), pregnant hypertensive and proteinuric women who did not meet criteria for severe preeclampsia (pHTN n = 21), and women with sPE (n = 17). RESULTS: There was no difference in gestational age at the time of enrollment among groups (median [range]: sPE: 31 [24-40], pHTN: 34 [16-40], P-CTR: 28 [7-39] wks). Urinary excretion of VEGF was significantly increased in sPE women compared with NP-CTR (P = .023), but did not differ among pregnant groups. Urinary PlGF levels were significantly increased in pregnant compared with nonpregnant women, but were decreased in all hypertensive women compared with healthy P-CTR (P < .001). Urinary sFlt-1 concentrations were significantly increased in women with sPE relative to all other groups (P < .001). pHTN women had higher sFlt-1 urinary output compared with P-CTR group (P = .001). A cutoff >2.1 in the ratio log [sFlt-1/PlGF] had 88.2% sensitivity and 100% specificity in differentiating women with sPE from normotensive controls. We also described that the log[sFlt-1/PlGF] ratio identified women with sPE better than proteinuria alone (P = .03). Our regression model revealed that uric acid correlated best with log[sFlt-1/PlGF] ratio (r = 0.628; P = .005). CONCLUSION: sPE is associated with increased urinary output of the antiangiogenic factor sFlt 1 and a decreased output of PlGF at the time of clinical manifestation, providing a rapid noninvasive screening of hypertensive women based on a sFlt/PlGF ratio. This ratio may be used as representation for severity of the disease, and appears to be superior to random urinary protein measurements. PMID- 15746667 TI - Prospective randomized clinical trial of inpatient cervical ripening with stepwise oral misoprostol vs vaginal misoprostol. AB - OBJECTIVE: The purpose of this study was to compare the efficacy and safety of stepwise oral misoprostol vs vaginal misoprostol for cervical ripening before induction of labor. STUDY DESIGN: Two hundred and four women between 32 to 42 weeks of gestation with an unfavorable cervix (Bishop score < or = 6) and an indication for labor induction were randomized to receive oral or vaginal misoprostol every 4 hours up to 4 doses. The oral misoprostol group received 50 microg initially followed by 100 microg in each subsequent dose. The vaginal group received 25 microg in each dose. The primary outcome was the interval from first misoprostol dose to delivery. Patient satisfaction and side effects were assessed by surveys completed after delivery. RESULTS: Ninety-three (45.6%) women received oral misoprostol; 111 (54.4%) received vaginal misoprostol. There was no difference in the average interval from the first dose of misoprostol to delivery in the oral (21.1 + 7.9 hrs) and vaginal (21.5 + 11.0 hrs, P = NS) misoprostol groups. The incidence of hyperstimulation in the oral group was 2.2% vs 5.4% in the vaginal group, P = NS. Eighteen patients in the oral group (19.4%) and 36 (32.4%) in the vaginal group underwent cesarean section (P < .05). This difference was attributed to better tolerance of more doses of misoprostol by the women in the oral group. There was no difference in side effects (nausea, vomiting, diarrhea, shivering) between groups. Fourteen percent of women in the vaginal group versus 7.5% in the oral group were dissatisfied with the use of misoprostol (P = NS). CONCLUSION: Stepwise oral misoprostol (50 microg followed by 100 microg) appears to be as effective as vaginal misoprostol (25 microg) for cervical ripening with a low incidence of hyperstimulation, no increase in side effects, a high rate of patient satisfaction, and is associated with a lower cesarean section rate. PMID- 15746668 TI - Effects of selective and nonselective PGE2 receptor agonists on cervical tensile strength and collagen organization and microstructure in the pregnant rat at term. AB - OBJECTIVE: The purpose of this study was to determine which of the 4 PGE2 receptors (EP1-EP4) is involved in cervical ripening in the rat, and to correlate its activity with changes in tensile strength and collagen microstructure. STUDY DESIGN: We assessed tensile strength after administration of selective and nonselective PGE2 receptor agonists. Quantification of collagen organization and microstructure was accomplished with polarized light microscopy and transmission electron microscopy. RESULTS: Selective agonists for EP1-3 did not produce significant differences when compared with each other or control animals. Significant differences in tensile strength, proportion of organized collagen, and microstructure were found between treatment and control animals with the nonselective receptor agonist (PGE2). This was taken as an indirect measure of EP4 activity. CONCLUSION: Changes in cervical collagen organization and microstructure are quantifiable and correlate with changes in tensile strength. These data implicate EP4 as the PGE2 receptor involved in producing these changes in the rat cervix. PMID- 15746669 TI - Recognition and treatment of irritable bowel syndrome among women with chronic pelvic pain. AB - OBJECTIVE: We sought to describe irritable bowel syndrome (IBS) treatment among women with chronic pelvic pain. STUDY DESIGN: We performed a cross-sectional study of new chronic pelvic pain patients between 1993 and 2000 (n = 987). IBS was defined by Rome I criteria. IBS treatment was defined as lower gastrointestinal drugs or referral. Analyses were descriptive and multivariable. RESULTS: IBS occurred in 35% of patients. In the highest quartile of pain, women with IBS were not more likely to have IBS treatment initiated. In the lowest three quarters of pain, women with IBS were 5.08 times more likely to have IBS treatment initiated. IBS was not diagnosed 40% of the time. IBS treatments were not recommended to 67% of patients with IBS. More than 35% of patients were prescribed narcotics. CONCLUSION: IBS is not consistently diagnosed and treated even in a pelvic pain clinic. Yet, treatment of IBS may reduce the overall abdominal pain of these patients. PMID- 15746670 TI - Laparoscopic radiofrequency thermal ablation: a new approach to symptomatic uterine myomas. AB - OBJECTIVE: The purpose of this study was to evaluate the feasibility and efficacy of laparoscopic radiofrequency ablation of uterine fibroids. STUDY DESIGN: Eighteen women with symptomatic intramural uterine myomas underwent radiofrequency ablation under laparoscopic guidance. Postoperative sonographic evaluations of the fibroids size were scheduled at 1, 3, 6 ,9, and 12 months. The impact of myoma-related symptoms on quality of life (QOL) was assessed using a validated questionnaire. RESULTS: The median number of myomas treated per patient was 1 (1-3). The median baseline volume of the dominant myoma was 67.2 cm 3 (14.8 332.8). No intraoperative or postoperative complications occurred. The median reductions in myomas volume were 41.5%, 59%, and 77% at 1, 3, and 6-months, follow-up evaluation, respectively. No further change in fibroid size was observed at 9 months and 1 year. A significant improvement in the symptoms score and QOL score was observed at 3 and 6 months, follow-up. CONCLUSION: In this pilot study, laparoscopic radiofrequency ablation successfully reduced fibroid symptoms and fibroid volume in short-term follow-up. Additional studies are needed before its efficacy and safety can be confirmed. PMID- 15746671 TI - The effect of vaginal candidiasis on the shedding of human immunodeficiency virus in cervicovaginal secretions. AB - OBJECTIVE: The purpose of this study was to evaluate the influence of symptomatic vulvovaginal candidiasis on the shedding of HIV-1 in cervicovaginal secretions of HIV-1-infected women. STUDY DESIGN: We obtained paired blood and cervicovaginal lavage samples from 66 HIV-infected women with symptomatic vulvovaginal candidiasis, and 249 HIV-infected control patients without genital infection. HIV 1 RNA in plasma, proviral HIV-1 DNA, HIV-1 RNA transcripts, and cell-free HIV-1 RNA in cervicovaginal secretions were quantitatively evaluated by competitive polymerase chain reaction (cPCR) and reverse transcriptase PCR (cRT-PCR). We used logistic regression on ordered data to assess the influence of vulvovaginal candidiasis on the HIV-1 load in cervicovaginal secretions adjusting for potential confounders. RESULTS: Overall, the amount of HIV-1 RNA in plasma was significantly correlated with HIV-1 DNA (Spearman rank 0.153 +/- 0.059, P = .006), HIV-1 RNA transcripts (Spearman rank 0.169 +/- 0.058, P = .003), and cell free HIV-1 RNA (Spearman rank 0.185 +/- 0.059, P = .001) load in cervicovaginal secretion. Forty-eight out of 182 (26.4%) patients who tested negative for HIV-1 RNA in plasma were positive for HIV-DNA in their cervicovaginal secretions. In logistic regression analysis vulvovaginal candidiasis was significantly associated with increasing loads of HIV-1 RNA transcripts (Odds ratio [OR] 1.97, 95% CI 1.09-3.57, P = .025) and cell free HIV-1 RNA (OR 2.03, 95% CI 1.10-3.73, P = .02) in cervicovaginal secretions. CONCLUSION: In HIV-infected women, vulvovaginal candidiasis is associated with an increased number of copies of cell associated and cell-free HIV-1 RNA in cervicovaginal secretions. PMID- 15746672 TI - Reliability of health-related quality-of-life measures 1 year after surgical procedures for pelvic floor disorders. AB - OBJECTIVE: The purpose of this study was to assess the reliability and validity of condition-specific health-related quality-of-life measures in women who are treated surgically for pelvic organ prolapse and urinary incontinence. STUDY DESIGN: The study used the cross-sectional telephone interview-based administration of a health-related quality-of-life measure, with a 2-week follow up interview for test-retest reliability. RESULTS: Initial and follow-up interviews were completed by 88 women (mean age, 65.7 +/- 11.6 years) approximately 1 year after surgical procedures. Condition-specific measures demonstrated acceptable reliability with test-retest correlation coefficients that approached or exceeded 0.6 and Cronbach's alpha that exceeded 0.8 in most domains. Validity was demonstrated with significant correlations of the urinary domains of the Pelvic Floor Distress Inventory and Pelvic Floor Impact Questionnaire, with continence defined by the medical, epidemiologic, and social aspects of aging and Hunskaar severity measures (all P < .001). CONCLUSION: The condition-specific health-related quality-of-life assessment is reliable and valid in women after surgical procedures for pelvic floor disorders. These findings support the inclusion of condition-specific health-related quality-of life measures in clinical trials for women with pelvic floor disorders. PMID- 15746673 TI - Pelvic floor morbidity at 3 years after instrumental delivery and cesarean delivery in the second stage of labor and the impact of a subsequent delivery. AB - OBJECTIVE: To compare pelvic floor symptoms at three years following instrumental delivery and cesarean section in the second stage of labor and to assess the impact of a subsequent delivery. STUDY DESIGN: We conducted a prospective cohort study of 393 women with term, singleton, cephalic pregnancies who required instrumental vaginal delivery in theatre or cesarean section at full dilatation between February 1999 and February 2000. 283 women (72%) returned postal questionnaires at three years. RESULTS: Urinary incontinence at three years post delivery was greater in the instrumental delivery group as compared to the cesarean section group (10.5% vs 2.0%), OR 5.37 (95% CI, 1.7, 27.9). There were no significant differences in ano-rectal or sexual symptoms between the two groups. Pelvic floor symptoms were similar for women delivered by cesarean section after a failed trial of instrumental delivery compared to immediate cesarean section. A subsequent delivery did not increase the risk of pelvic floor symptoms at three years in either group. CONCLUSION: An increased risk of urinary incontinence persists up to three years following instrumental vaginal delivery compared to cesarean section in the second stage of labor. However, pelvic floor symptoms are not exacerbated by a subsequent delivery. PMID- 15746675 TI - Comparison of 2-dimensional and 3-dimensional power-Doppler imaging in complex adnexal masses for the prediction of ovarian cancer. AB - OBJECTIVE: The purpose of this study was to compare 2-dimensional and 3 dimensional power-Doppler imaging diagnostic performance for the prediction of ovarian cancer in complex adnexal masses. STUDY DESIGN: Sixty-nine complex adnexal masses in 60 women (mean age, 48.4 years [range, 17-82 years]) were evaluated by 2-dimensional and 3-dimensional power-Doppler imaging for differentiating benign from malignant masses. Complex adnexal mass was defined in the presence of at least 1 of the following features: solid areas, thick papillary projections, thick septa, or purely solid echogenicity. One examiner performed 2-dimensional power-Doppler imaging, and a second examiner performed 3 dimensional power-Doppler imaging. All masses were removed surgically, and definitive diagnosis was obtained. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for both techniques. RESULTS: Forty-five tumors (65.2%) were proved to be malignant, and 24 tumors (34.8%) were proved to be benign. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for 2-dimensional power Doppler imaging were 97.8%, 87.5%, 93.6%, 95.5%, and 94.2%, respectively. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for 3-dimensional power-Doppler imaging were 97.8%, 79.2%, 89.9%, 95%, and 91.3 % respectively. There were no statistical differences in sensitivity and specificity (McNemar test: P = .250) CONCLUSION: Three dimensional power-Doppler imaging did not have a better diagnostic performance than 2-dimensional power-Doppler imaging for the discrimination of benign from malignant complex adnexal masses. PMID- 15746674 TI - Pelvic Organ Support Study (POSST): the distribution, clinical definition, and epidemiologic condition of pelvic organ support defects. AB - OBJECTIVE: The purpose of this study was to describe the distribution of pelvic organ support in a gynecologic clinic population to define the clinical disease state of pelvic organ prolapse and to analyze its epidemiologic condition. STUDY DESIGN: This was a multicenter observational study. Subjects who were seen at outpatient gynecology clinics who required an annual gynecologic examination underwent a pelvic organ prolapse quantification examination and completed a prolapse symptom questionnaire. Receiver operator characteristic curves were used to define pelvic organ prolapse with the use of symptoms and pelvic organ prolapse quantification examination measures. Standard age-adjusted univariate and multivariate logistic regression analysis were used to evaluate various relationships. RESULTS: The population consisted of 1004 women who were aged 18 to 83 years. The prevalence of pelvic organ prolapse quantification stages was 24% (stage 0), 38% (stage 1), 35% (stage 2), and 2% (stage 3). The definition of pelvic organ prolapse that was determined by the receiver operator characteristic curve was the leading edge of their vaginal wall that was -0.5 cm above the hymenal remnants. Multivariate analysis revealed age, Hispanic race, increasing body mass index, and the increasing weight of the vaginally delivered fetus as risk factors for pelvic organ prolapse, as defined in this population. CONCLUSION: The results from this population suggest that there is a bell-shaped distribution of pelvic organ support in a gynecologic clinic population. Advancing age, Hispanic race, increasing body mass index, and the increasing weight of the vaginally delivered fetus have the strongest correlations with prolapse. PMID- 15746677 TI - Expression of cyclooxygenase-2 in advanced stage ovarian serous carcinoma: correlation with tumor cell proliferation, apoptosis, angiogenesis, and survival. AB - OBJECTIVE: Cyclo-oxygenase-2 seems to be involved at various steps in the processes of tumor progression. The objective of this study was to examine the relationship between cyclo-oxygenase-2 expression and tumor proliferation, apoptosis and angiogenesis in patients with advanced stage high-grade ovarian carcinoma. STUDY DESIGN: Specimens from 118 patients with high-grade and advanced stage (III, IV) serous ovarian carcinoma were evaluated by immunohistochemistry for cyclo-oxygenase-2, Ki-67, vascular endothelial growth factor, and bcl-2 expression. Tumor microvessel density was assessed with CD34 immunostaining. We investigated the relationships between cyclo-oxygenase-2 expression and clinicopathologic characteristics, tumor angiogenesis (tumor microvessel density and vascular endothelial growth factor expression), and tumor proliferation and apoptosis. The effect of cyclooxygenase-2 expression on patient survival was determined. RESULTS: There was a significant positive correlation between cyclo oxygenase-2 expression in tumor cells and markers of tumor proliferation and angiogenesis. In univariate survival analysis, high cyclo-oxygenase-2 and high Ki 67 expression showed a significant impact of on patient survival (P < .001). In multivariate regression analysis, only Ki-67 expression retained its significance as an independent poor prognostic factor (death hazard ratio, 2.0; 95% CI, 1.2 3.3; P < .001). CONCLUSION: Expression of cyclo-oxygenase-2 correlates with tumor proliferation and tumor angiogenesis but not with apoptotic markers (bcl-2 expression) in high-grade, advanced-stage serous ovarian carcinoma. PMID- 15746676 TI - Racial differences in the overexpression of epidermal growth factor type II receptor (HER2/neu): a major prognostic indicator in uterine serous papillary cancer. AB - OBJECTIVE: A difference in survival rates between black and white patients with cancer of the corpus uteri is well established. This study was conducted to determine whether the overexpression of HER2/neu oncogene is associated with poor outcome in uterine serous papillary endometrial cancer, which is a highly aggressive variant of endometrial cancer, and whether a racial difference in the frequency of HER2/neu overexpression may contribute to the disparity in endometrial cancer survival. STUDY DESIGN: Immunohistochemical evaluation was used to examine HER2/neu expression in paraffin blocks from 27 women with stage IA to IV uterine serous papillary endometrial cancer. Univariable analysis was performed and followed by multivariable analysis with Cox's proportional hazard model to evaluate whether HER2/neu expression was associated with poor outcome in uterine serous papillary endometrial cancer. RESULTS: Black patients tended to be younger (P = .02) and have higher HER2/neu expression than white patients (trend P = .02). Seven of 10 black patients (70%) showed heavy (3+) expression, compared with 4 of 17 white patients (24%; P = .04). The association of heavy HER2/neu expression with race persisted after age was controlled through stratification (P = .05). Earlier deaths from uterine serous papillary endometrial cancer were seen among heavy HER2/neu expressers (P = .002), black patients (P = .04), and patients < or = 65 years old (P = .04). However, multivariate Cox regression showed that short survival was associated significantly with heavy HER2/neu expression (P = .02) but not with age (P = .07) or race (P = .35), which indicates that HER2/neu expression accounted for much of the race disparity in survival in this patient population. CONCLUSION: Overexpression of HER2/neu in uterine serous papillary endometrial cancer is an independent variable that is associated with poor outcome, occurs more frequently in black women, and may contribute to racial disparity in survival. HER2/neu expression may guide clinical treatment of patients with uterine serous papillary endometrial cancer and may have implications for the implementation of novel treatment strategies. PMID- 15746678 TI - Catastrophizing labor pain compromises later maternity adjustments. AB - OBJECTIVE: The purpose of this study was to evaluate the impact of labor pain intensity and labor pain catastrophizing on maternity blues and postpartum social functioning. STUDY DESIGN: Pain intensity and pain catastrophizing were assessed in 89 women in active labor before the administration of analgesia. Both these measures were assessed again retrospectively 2 days after delivery in 82 women who had a spontaneous vaginal delivery. Women also filled out the Edinburgh Postnatal Depression Scale. Six weeks later women completed the social functioning domain of the short form SF36 health survey. RESULTS: Pain catastrophizing during labor significantly predicted both maternity blues (P = .001) and postpartum social functioning (P = .001) when being controlled for maternal age and education, parity, type of analgesia, and labor pain intensity. Low level of education and younger age also contributed to the prediction of maternity blues and social functioning. CONCLUSION: Labor pain catastrophizing rather than labor pain intensity predicts postpartum maternal adjustments. PMID- 15746679 TI - Population-based trends and correlates of maternal overweight and obesity, Utah 1991-2001. AB - OBJECTIVE: This study aims to identify recent population-based trends in maternal overweight and obesity and adverse outcomes. STUDY DESIGN: Statewide retrospective cohort study of birth certificate data for live singleton births to women in Utah between 1991 and 2001. RESULTS: Prepregnancy overweight and obesity increased from 25.1% in 1991 to 35.2% in 2001, a 40.2% increase (prevalence ratio [PR] 1.40 [1.37-1.43]), whereas maternal obesity at delivery rose 36.2% from 28.7% to 39.1% (PR 1.36 [1.33-1.39]). The attributable fraction of cesarean delivery in overweight and obese women was 0.388 (0.369-0.407). Statewide, among all women having a cesarean delivery in 2001, 1 in 7 is attributable to overweight and obesity. CONCLUSION: This is the first state-wide analysis of maternal obesity trends demonstrating a significant increase in maternal overweight and obesity. Overweight and obese women are at increased risk of cesarean delivery, preeclampsia, eclampsia, dystocia, and macrosomia, risks that increase as the body mass index rises. PMID- 15746680 TI - Neonatal death and morbidity in vertex-nonvertex second twins according to mode of delivery and birth weight. AB - OBJECTIVE: The purpose of this study was to assess the risk of neonatal death and morbidity in vertex-nonvertex second twins according to the mode of delivery and birth weight. STUDY DESIGN: Data from a retrospective cohort study that was based on all twin births in the United States (1995-1997) were used. RESULTS: A total of 15,185 vertex-nonvertex second twins were classified into 3 groups: (1) both twins were delivered by cesarean delivery (37.7%), (2) both twins were delivered vaginally (46.8%), and (3) the second twin was delivered by cesarean delivery after vaginal delivery of the first twin (15.5%). The risk of asphyxia-related neonatal deaths and morbidity was increased in the group in which both twins were delivered vaginally and the group in which both twins were delivered by cesarean delivery. The increase in neonatal death in the group in which both twins were delivered vaginally was stronger in the birth weight of < 1500 g. In contrast, in the group in which both twins were delivered vaginally and the group in which the second twin was delivered by cesarean delivery after the first twin was delivered vaginally, the increase in neonatal morbidity was greater in the group in which the birth weight was 1500 to 4000 g. CONCLUSION: The risk of neonatal death and morbidity in second-born twins is higher in the group in which both twins were delivered vaginally and the group in which the second twin was delivered by cesarean delivery after the first twin was delivered vaginally compared with the group in which both twins were delivered by cesarean delivery. PMID- 15746681 TI - Maternal age and the likelihood of a maternal request for cesarean delivery: a 5 year population-based study. AB - OBJECTIVE: The purpose of this study was to examine associations between maternal age and maternal request cesarean deliveries. STUDY DESIGN: Five-year population based data from Taiwan (1997-2001) that covered 904,657 singleton deliveries without a clinical indication for cesarean delivery that were judged by the attending physician were subjected to multiple logistic regression, year-wise, to examine the association of maternal age with request cesarean delivery, adjusted for health care institutional characteristics. RESULTS: Request cesarean delivery rates steadily increased over the study period within each age group, disproportionately so among the 34+ age group. Women aged < 25 years were less likely than women aged 25 to 34 years (reference group) to request a cesarean delivery (odds ratio range, 0.67-0.88) and women aged 34+ were more likely than the reference group to have a request cesarean delivery (odds ratio range, 1.96 2.01), adjusted for health care institutional characteristics. CONCLUSION: Population-based data confirms the expectancy that request cesarean delivery propensity increases with maternal age. PMID- 15746682 TI - Second trimester abortion using isosorbide mononitrate in addition to gemeprost compared with gemeprost alone: a double-blind randomized, placebo-controlled multicenter trial. AB - OBJECTIVE: We aimed to determine whether second-trimester abortion using isosorbide mononitrate (IMN) in addition to gemeprost is more effective and reduces side effects compared with gemeprost alone. STUDY DESIGN: Eighty women who were age 13 to 23 weeks' gestation were randomly assigned to receive per vaginam either IMN 40 mg (group 1, 40 women) or placebo (group 2, 40 women) in addition to gemeprost 1 mg up to 3 times daily 3 hours apart for 2 days. Analysis of variance, a chi 2 test, and a multivariate analysis were performed. RESULTS: Of the 72 women analyzed, 68% (group 1) and 38% (group 2) underwent abortion within day 1 (P < .05). However, group 1 was associated with more headache (18% of women) 3 hours after induction compared to group 2 (0% of women, P = .038). CONCLUSION: IMN in addition to gemeprost is effective for second-trimester abortion, but is associated with more headache compared with gemeprost alone. PMID- 15746683 TI - Risk factors for neonatal mortality among extremely-low-birth-weight infants. AB - OBJECTIVE: The purpose of this study was to examine characteristics associated with neonatal mortality among extremely low-birth-weight infants (< or = 1000 g). STUDY DESIGN: A population-based, case-control study using linked Missouri birth and death certificates from 1989 to 1997 was conducted. Cases (n = 835) were defined as extremely low-birth-weight infants that died within 28 days of birth. Controls (n = 907) were randomly selected from extremely low-birth-weight infants that were alive at 1 year and were frequency matched to subjects by birth year and birth weight. RESULTS: Infants born with severe congenital anomalies and at the youngest gestational ages were at greatest risk for neonatal mortality. Other significant risk factors included maternal age (< 18 and > 34 years), vaginal delivery, nontertiary hospital care, malpresentation, male gender, and small for gestational age. Black race and preeclampsia were protective against early death. CONCLUSIONS: The risk of neonatal mortality among extremely low-birth-weight infants was associated with several maternal, infant, and obstetric factors, some of which may be preventable. PMID- 15746684 TI - Birth simulator: reliability of transvaginal assessment of fetal head station as defined by the American College of Obstetricians and Gynecologists classification. AB - OBJECTIVE: This study was undertaken to investigate the reliability of transvaginal assessment of fetal head station by using a newly designed birth simulator. STUDY DESIGN: This prospective study involved 32 residents and 25 attending physicians. Each operator was given all 11 possible fetal stations in random order. A fetal head mannequin was placed in 1 of the 11 American College of Obstetricians and Gynecologists (ACOG) stations (-5 to +5) in a birth simulator equipped with real-time miniaturized sensor. The operator then determined head position clinically using the ACOG classification. Head position was described as: (1) "engaged" or "nonengaged" (engagement code); (2) "high," "mid," "low," or "outlet" (group code); and (3) according to the 11 ACOG ischial spine stations (numerical code). Errors were defined as differences between the stations given by the sensor and by the operator. We determined the error rates for the 3 codes. RESULTS: "Numerical" errors occurred in 50% to 88% of cases for residents and in 36% to 80% of cases for attending physicians, depending on the position. The mean "group" error was 30% (95% CI 25%-35%) for residents and 34% (95% CI 27%-41%) for attending physicians. In most cases (87.5% for residents and 66.8% for attending physicians) of misdiagnosis of "high" station, the "mid" station was retained. Residents and attending physicians made an average of 12% of "engagement" errors, equally distributed between false diagnosis of engagement and nonengagement. CONCLUSION: Our results show that transvaginal assessment of fetal head station is poorly reliable, meaning clinical training should be promoted. The choice not to perform vaginal delivery when the fetus is in the "mid" position strongly decreases the risk of applying instruments on an undiagnosed "high" station. Conversely, obstetricians who perform only "low" operative vaginal deliveries also deliver unrecognized "mid" station fetuses. Therefore, residency programs should offer training in "mid" pelvic operative vaginal deliveries. Birth simulators could be used in training programs. PMID- 15746685 TI - Factors predicting severe perineal trauma during childbirth: role of forceps delivery routinely combined with mediolateral episiotomy. AB - OBJECTIVE: Anal sphincter injury and its sequelae are a recognized complication of vaginal childbirth. The aim of the present study was to identify risk factors for third- and fourth-degree perineal tears in patients undergoing either spontaneous or vaginal-assisted delivery by forceps routinely combined with mediolateral episiotomy. STUDY DESIGN: We retrospectively reviewed 5377 vaginal deliveries based on the analysis of the obstetric database and patient records of our department during a 5-year period from 1999 to 2003. Cases and control subjects were chosen randomly and patients' records were reviewed for the following variables: maternal age, parity, gestational age, tobacco use, gestational diabetes or pregnancy-induced hypertension, use of peridural anesthesia, duration of first and second stages of labor, use of mediolateral episiotomy, forceps combined with mediolateral episiotomy, induction of labor, infant head diameter, shoulder circumference, and birth weight. RESULTS: Of 5044 spontaneous vaginal deliveries 32 (0.6%) and of 333 assisted vaginal deliveries 14 (4.2%) patients sustained a perineal defect involving the external sphincter. An univariate analysis of these 46 cases and 155 randomly selected control subjects showed that low parity (P = .003; Mann-Whitney U test), prolonged first and second stages of labor (P = .001, P = .001), high birth weight (P = .031), episiotomy (P = .004; Fisher exact test), and forceps delivery (P = .002) increased the risk for sphincter damage. In multivariate regression models, only high birth weight (P = .004; odds ratio [OR] 1.68, 1.18-2.41, 95% confidence interval [CI]), and forceps delivery combined with mediolateral episiotomies (P < .001; OR 5.62, 2.16-14.62, 95% CI) proved to be independent risk factors. There was a statistical significant interaction of birth weight and head circumference (P = .012; OR 0.99, 0.98-0.99, 95% CI). Although the use of episiotomy conferred an increased risk toward a higher likelihood of severe perineal trauma, it did not reach statistical significance (P = .06; OR 2.15, 0.97-4.76, 95% CI). CONCLUSIONS: In consistence with previous reports, women who are vaginally delivered of a large infant are at a high risk for sphincter damage. Although the rate of these complications was surprisingly low in vaginally assisted childbirth, the use of forceps, even if routinely combined with mediolateral episiotomy, should be minimized whenever possible. PMID- 15746686 TI - The Preterm Prediction Study: association between maternal body mass index and spontaneous and indicated preterm birth. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between prepregnancy maternal body mass index and spontaneous preterm birth and indicated preterm birth. STUDY DESIGN: This was a secondary analysis of the Maternal-Fetal Medicine Units Network, Preterm Prediction study. Patients were classified into categories that were based on their body mass index. Rates of indicated and spontaneous preterm birth were compared. RESULTS: Five hundred ninety-seven (20.5%) of 2910 women were obese. Obese women had fewer spontaneous preterm births at < 37 weeks of gestation (6.2% vs 11.2%; P < .001) and at < 34 weeks of gestation (1.5% vs 3.5%; P = .012). Women with a body mass index of < 19 kg/m2 had 16.6% spontaneous preterm birth, with a body mass index of 19 to 24.9 kg/m 2 had 11.3% spontaneous preterm birth, with a body mass index of 25 to 29.9 kg/m2 had 8.1% spontaneous preterm birth, with a body mass index of 30 to 34.9 kg/m2 had 7.1% spontaneous preterm birth, and with a body mass index of > or = 35 kg/m2 had 5.2% spontaneous preterm birth (P < .0001). Indicated delivery was responsible for an increasing proportion of preterm births with increasing body mass index (P = .001). Obese women had lower rates of cervical length < 25 mm (5% vs 8%; P = .012). Multivariable regression analysis confirmed a lower rate of spontaneous preterm birth in obese gravid women (odds ratio, 0.57; 95% CI, 0.39 0.83; P = .003). CONCLUSION: Obesity before pregnancy is associated with a lower rate of spontaneous preterm birth. PMID- 15746687 TI - Assessment of cervical antibody concentrations fails to enhance the value of cervical length as a predictor of preterm delivery. AB - OBJECTIVE: The purpose of this study was to determine if cervical fluid antibody concentrations can enhance the value of cervical length in predicting risk of preterm delivery. STUDY DESIGN: We obtained cervical fluid samples with preweighed cellulose wicks from a prospective cohort of women 23 to 32 weeks' gestation with signs and symptoms of preterm labor and intact membranes. Total immunoglobulin A and G (IgA and IgG) concentrations were determined by enzyme linked immunosorbent assay. Bacterial vaginosis was diagnosed by Gram stain, and cervical length was measured with endovaginal ultrasound. RESULTS: For subjects with term (n = 77) and preterm (n = 24) deliveries, median IgA and IgG concentrations were 736 vs 643 microg/mL (P = .33) and 1528 vs 1769 microg/mL (P = .85). For subjects with normal flora (n = 71), intermediate flora (n = 14), and bacterial vaginosis (n = 16), median IgA and IgG concentrations were 717, 624, and 774 microg/mL (P = .90) and 1383, 1553, and 2731 microg/mL (P = .02). In a forward stepwise logistic regression model, cervical length was the only factor associated with preterm delivery (P < .001). CONCLUSION: Measuring the concentrations of IgA and IgG in cervical fluid does not enhance the value of cervical length in predicting risk of preterm delivery. PMID- 15746688 TI - Prenatal cardiovascular manifestations in the twin-to-twin transfusion syndrome recipients and the impact of therapeutic amnioreduction. AB - OBJECTIVE: We evaluated the cardiovascular pathologic condition in the recipient twin in twin-to-twin transfusion syndrome and the influence of amnioreduction. STUDY DESIGN: Fetal echocardiograms and medical records of 54 pregnancies that were complicated by twin-to-twin transfusion syndrome were reviewed. Recipient twin right and left ventricular wall thickness, diameters, systolic and diastolic function, valve regurgitation, and structural cardiac defects were assessed at examination and after amnioreduction. RESULTS: At examination (n = 28 pregnancies), cardiomegaly because of right ventricular and/or left ventricular hypertrophy was observed in 58% of recipient twins, and biventricular hypertrophy was observed in 33% of recipient twins, without ventricular dilation. Biventricular diastolic dysfunction was present in two thirds of recipient twins, and right ventricular systolic dysfunction and significant atrioventricular valve regurgitation was observed in one third of recipient twins. Serial assessment (n = 21 pregnancies) revealed progressive biventricular hypertrophy and right ventricular systolic and biventricular diastolic dysfunction in most recipient twins. Steeper progression of hypertrophy, diastolic dysfunction, and structural or functional right ventricular outflow disease (20% incidence) were associated with an increased perinatal mortality rate. CONCLUSION: In twin-to-twin transfusion syndrome, the recipient twin has progressive biventricular hypertrophy with predominant right ventricular systolic and biventricular diastolic dysfunction. Despite amnioreduction, the cardiovascular disease persists and even progresses in many recipient twins. PMID- 15746689 TI - Tocolytic effect of a Rho-kinase inhibitor in a mouse model of lipopolysaccharide induced preterm delivery. AB - OBJECTIVE: The small guanosine triphosphatase RhoA/Rho-kinase cascade has been implicated in uterine contraction. Our purpose was to evaluate the tocolytic effect of a Rho-kinase inhibitor, Y-27632, in lipopolysaccharide-induced preterm delivery in mice. STUDY DESIGN: We used an animal model of lipopolysaccharide induced preterm delivery in C3H/HeN x B6D2F1 pregnant mice. Y-27632 was delivered continuously through an osmotic pump that was implanted into the peritoneal cavity 6 hours before lipopolysaccharide treatment. The primary outcome was the preterm delivery rate. To further study the possible involvement of this cascade in lipopolysaccharide-induced preterm delivery, we determined the effect of lipopolysaccharide and prostaglandin F2alpha on RhoA activation in mouse myometrial cells and uterine smooth muscle tissues. RESULTS: The rate of preterm delivery for lipopolysaccharide-treated animals was 94.4%. The administration of Y-27632 (1 or 10 mg/kg/d) significantly reduced the preterm delivery rate to 61.1% or 15.8%, respectively. The level of guanosine triphosphate-bound RhoA was increased after the addition of lipopolysaccharide or prostaglandin F2alpha. CONCLUSION: The RhoA/Rho-kinase cascade is involved in lipopolysaccharide-induced preterm delivery, which suggests that Rho-kinase could be used as a new therapeutic target for the prevention of preterm labor. PMID- 15746690 TI - The cost of twin pregnancy: maternal and neonatal factors. AB - OBJECTIVE: The purpose of this study was to evaluate factors affecting birth charges in twin pregnancies. STUDY DESIGN: Clinical and financial data were obtained on 1486 twin pregnancies delivered between 1995 to 2002 at medical centers in Maryland, Florida, Michigan, and South Carolina. Maternal and neonatal length of stay (LOS) and charges were modeled by gestational age and other risk factors using a general linear model. RESULTS: Maternal and infant birth admission LOS and charges increased significantly with a decline in gestational age. Maternal LOS and charges were also significantly increased by cesarean delivery and preeclampsia. Newborn LOS and charges increased significantly by monochorionicity and slowed growth between 20 to 28 weeks. For mother and infants, the shortest LOS and lowest birth charges were at 37 to 38 weeks. CONCLUSION: These findings reflect the substantial maternal and neonatal morbidity associated with twin pregnancies, and demonstrate that 37 to 38 weeks is their optimal gestation. PMID- 15746691 TI - Association between vaginal 70-kd heat shock protein, interleukin-1 receptor antagonist, and microbial flora in mid trimester pregnant women. AB - OBJECTIVE: This study investigated the association among the inducible 70-kd heat shock protein, cytokines, and microbial flora in the vagina in mid trimester pregnant women and subsequent preterm delivery. STUDY DESIGN: Vaginal samples from 205 pregnant women, which were collected at 18 to 22 weeks of gestation, were analyzed for qualitative and quantitative vaginal microflora and for 70-kd heat shock protein, interleukin-1beta, interleukin-1 receptor antagonist, and tumor necrosis factor-alpha by enzyme-linked immunosorbent assay. Pregnancy outcome data were obtained subsequently. RESULTS: The 70-kd heat shock protein was detected in 38 vaginal samples (18.5%). Its presence was associated with elevated vaginal pH, a diagnosis of bacterial vaginosis, and elevated interleukin 1 receptor antagonist levels (P < .001). Among women with bacterial vaginosis, 70 kd heat shock protein-positive subjects had a >80% increase in median vaginal concentration of interleukin-1 receptor antagonist (P < .05). CONCLUSION: Vaginal 70-kd heat shock protein expression is associated with the down-regulation of the proinflammatory immune response to abnormal vaginal flora in mid trimester pregnant women. PMID- 15746692 TI - Aspirin use during early pregnancy and the risk of congenital abnormalities: a population-based case-control study. AB - It is still controversial whether aspirin use during pregnancy increases the risk of certain congenital abnormalities (in particular, neural tube defects, gastroschisis, and cleft lip +/- palate). We examined the risk of selected congenital abnormalities in a large case-control dataset from Hungary and found no increased risk of such congenital abnormalities. PMID- 15746693 TI - Dose response of RU486 in a novel rabbit model of noninfectious preterm birth: comparative efficacy of 3 routes of administration. AB - OBJECTIVE: The purpose of this study was to examine whether the pregnant rabbit model can be used as a viable model for the study of non-infection-mediated preterm birth. STUDY DESIGN: Timed pregnant New Zealand rabbits were injected with a single dose of RU486 on day 22 of gestation. Three doses (50 mg, 75 mg, and 100 mg) were administered intramuscularly, intraperitoneally, or subcutaneously. The rabbits were monitored for preterm delivery. Progesterone, cortisol, and cytokine levels were examined before the induction and after delivery. Uterine and cervical progesterone, cortisol, and cytokine levels were determined after delivery. RESULTS: RU486 resulted in 100% preterm delivery in all doses and modes of administration, compared with 0% of controls. Intramuscular administration appeared to generate the most favorable preterm delivery time. Rabbits that received 100 mg RU486 intramuscularly showed significantly decreased serum progesterone levels and uterine progesterone levels, compared with 100 mg subcutaneously and intraperitoneally. CONCLUSION: RU486 that was administered intramuscularly appears to be a potent and effective method for inducing preterm birth. This model of hormonally mediated preterm birth might serve as a useful model for the investigation of the possible mechanisms of preterm labor. PMID- 15746694 TI - Pregnancy outcome of women exposed to bupropion during pregnancy: a prospective comparative study. AB - OBJECTIVE: Bupropion was developed for the treatment of depression, but subsequently was found to be effective for smoking cessation. To date, there are no prospective comparative studies examining its safety in pregnancy. The primary objective was to determine whether bupropion increases the risks for major malformations above baseline. The secondary objective was to examine the rates of live births, stillbirths, spontaneous and therapeutic abortions, mean birth weight, and gestational age at birth. STUDY DESIGN: Women who were pregnant or planning a pregnancy and taking bupropion were enrolled in the study. Follow-up of pregnancy outcome was carried out between 4 months and 1 year after delivery. Three comparisons were carried out: 1) women exposed to bupropion vs a nonteratogen group; 2) those taking for depression vs other antidepressants, vs a nonteratogen group; 3) spontaneous abortions were compared between those taking for depression, vs another antidepressant group vs a nonteratogen group. RESULTS: We completed follow-up on 136 women exposed to bupropion during the first trimester of pregnancy. There were (105) live births, no major malformations, the mean birth weight was (3450g), the mean gestational age at delivery was (40 weeks), the number of spontaneous abortions was 20, there were 10 therapeutic abortions, there was 1 stillbirth, and 1 neonatal death. There were no statistically significant differences between any of the end points we examined between the exposed and comparison groups, with the exception of significantly more spontaneous abortions in the bupropion group (P = .009). CONCLUSION: These results suggest that bupropion does not increase the rates of major malformation above baseline. The higher rates of spontaneous abortions are similar to other studies examining the safety of antidepressants during pregnancy. PMID- 15746695 TI - Reference ranges for serial measurements of umbilical artery Doppler indices in the second half of pregnancy. AB - OBJECTIVE: The purpose of this study was to construct new reference ranges for serial measurements of commonly used umbilical artery Doppler indices (pulsatility index, resistance index, and systolic:diastolic ratio). STUDY DESIGN: This was a prospective longitudinal study of the umbilical artery Doppler indices that were obtained serially at the free-loop of umbilical cord at 4-week intervals at 19 to 42 weeks of gestation in 130 low-risk singleton pregnancies. A total of 513 observations were used to construct the reference ranges with the use of multilevel modeling. RESULTS: Longitudinally established percentiles of Doppler indices from the present study show a continuous reduction throughout the second half of pregnancy without any plateau or increase near term, as reported previously. There was a significant negative association between Doppler indices and placental weight and neonatal birth weight, but not with gender. The intraobserver coefficients of variation for the umbilical artery pulsatility index, resistance index, and systolic:diastolic ratio were 10.5%, 6.8 %, and 13.0 %, respectively. CONCLUSION: New reference ranges for umbilical artery Doppler indices that are based on longitudinal observations appear to be slightly different from cross-sectional studies and are more appropriate for serial evaluation of fetal hemodynamics. PMID- 15746696 TI - Maternal gestational protein-calorie restriction decreases the number of glomeruli and causes glomerular hypertrophy in adult hypertensive rats. AB - OBJECTIVE: This work analyzed the renal function and structure in offspring rats that were submitted to maternal protein-calorie restriction during prenatal or lactation periods. STUDY DESIGN: Kidneys from adult offspring were studied. Animals from mothers that were submitted to food restriction were separated in 3 groups: control, prenatal restriction, and lactation restriction. Blood pressure, microalbuminuria, and glomerular filtration rate were determined. Kidney cortical remodeling was analyzed with stereology; volume-weighted glomerular volume and the number of glomeruli were estimated. RESULTS: Adult prenatal restriction offspring showed enhanced microalbuminuria, decreased glomerular filtration rate, and hypertension; their kidneys showed a smaller number of hypertrophied glomeruli than control and lactation restriction animals. CONCLUSION: Maternal prenatal protein-calorie restriction in rats causes kidney disease in adult offspring, which is characterized by hypertension and renal dysfunction and suggests secondary kidney remodeling because of an impairment of glomerulogenesis. PMID- 15746697 TI - Antihypertensive effects of flutamide in rats that are exposed to a low-protein diet in utero. AB - OBJECTIVE: We investigated whether gestational age of in utero low-protein diet played a role in the subsequent development of adult hypertension and whether it is gender dependent and examined whether flutamide (a specific, nonsteroidal competitive antagonist of the androgen receptor) reduces blood pressure in rat offspring that are exposed to in utero low-protein diet (6%). STUDY DESIGN: Pregnant rats were fed either with 20% protein (control) or 6% protein (low protein diet) from day 1 or day 12 of gestation. Fetoplacental weights and mortality rates of pups were assessed. Systolic blood pressure, mean arterial blood pressure, and circulatory hormone levels in offspring were determined. In addition, male and female hypertensive offspring were treated with flutamide, and their blood pressure was monitored. RESULTS: After delivery, pup weights were reduced, and pup mortality rates increased in the low-protein diet-day 1 group. Systolic blood pressure and mean arterial blood pressure were elevated in low protein diet-day 1 males and females and low-protein diet-day 2 males. Significant (P < .05) reduction in blood pressure was achieved with flutamide in low-protein diet-day 1 females. Serum estradiol levels were decreased (P < .05) in low-protein diet-day 1 females; flutamide attenuated this effect. CONCLUSION: The day of in utero insult by low-protein diet is critical in the induction of adult hypertension; the severity is gender dependent. Flutamide was found to protect against hypertension only in females. PMID- 15746698 TI - Placental vascular disease and toll-like receptor 4 gene expression. AB - OBJECTIVE: Vascular disease in the placenta, which is identified by the study of umbilical artery Doppler flow velocity waveforms, is associated with endothelial cell activation and a proinflammatory cytokine response in the villous placental circulation. We studied toll-like receptor 4 expression (the ligand is lipopolysaccharide) to examine whether infection may cause these inflammatory components of placental vascular disease through an innate immune response. STUDY DESIGN: Microvessel endothelial cells were isolated from human placentae with collagenase digestion and then extracted with Dynabeads that were coated with monoclonal antibody against CD31. We studied 13 placentae from normal pregnancies that were delivered at term and 15 pregnancies with umbilical placental vascular disease that was defined by an abnormal umbilical artery Doppler study. We extracted RNA from the isolated endothelial cells. The messenger RNA expression of toll-like receptor 4 production was assessed by reverse transcriptase polymerase chain reaction and factored relative to the glyceraldehyde-3-phosphate dehydrogenase and 18S ribosomal RNA genes. RESULTS: Microvessel endothelial cells from placental villi with placental vascular disease showed up-regulation of toll like receptor 4 expression (toll-like receptor 4/18S, 1.92 +/- 0.37 vs 0.99 +/- 0.19; P < .05; toll-like receptor 4/glyceraldehyde-3-phosphate dehydrogenase, 2.20 +/- 0.36 vs 1.25 +/- 0.22; P < .05) in comparison with normal pregnancy. CONCLUSION: Up-regulation of toll-like receptor 4 gene in the endothelium of the placental villi is present in placental vascular disease, which may result from exposure of this endothelium to the toll-like receptor 4 ligand lipopolysaccharide in vivo. Directly extracted endothelial cells were used to avoid the possibility for change in behavior in tissue culture. We conclude that Gram-negative infection and lipopolysaccharide stimulation may cause placental vascular disease. PMID- 15746699 TI - Aberrant patterns of cellular communication in diabetes-induced embryopathy in rats: II, apoptotic pathways. AB - OBJECTIVE: The objective was to test the hypothesis that hyperglycemia-induced injury of yolk sac cell membranes is associated with disruption of cellular apoptotic signaling pathways. STUDY DESIGN: Pregnant rats were induced to become diabetic by injection of streptozotocin. Fourteen normal control and 24 diabetic rats were killed on day 12 of gestation. Yolk sac membranes in 3 conceptus groups (nondiabetic, diabetic with normal, or diabetic with malformed conceptus) were collected for study. DNA was extracted from yolk sac cells and assayed for fragmentation by using gel electrophoresis, which indicates apoptosis. Protein expression was evaluated by Western blot assays. Statistical analyses were performed with the Student t -test. RESULTS: The level of phosphorylated Akt was significantly decreased, whereas that of the proapoptotic protein Bax was increased. These changes were correlated with the presence of DNA fragmentation in yolk sac cells of the diabetic malformed conceptuses. CONCLUSION: Hyperglycemia-induced embryopathy involves apoptosis, during which the expression of proapoptotic protein Bax is upregulated and the activity of the cell-survival factor, Akt kinase, is decreased in yolk sac cells. These observations suggest that hyperglycemia of maternal diabetes triggers apoptotic signaling pathways and inhibits cell survival pathways, leading to embryonic malformations. PMID- 15746700 TI - Impaired KATP channel function in the fetoplacental circulation of patients with type 1 diabetes mellitus. AB - OBJECTIVE: The increased perinatal morbidity in diabetes may be partly related to vascular dysfunction. Because potassium channels play an important role in the regulation of vascular tone, this study explores the impact of diabetes on potassium channel function in the fetoplacental vascular bed. STUDY DESIGN: Vascular potassium channel function was investigated by ex vivo dual perfusion of isolated placental cotyledons (n = 47). Appropriate control experiments were carried out to exclude nonspecific effects. RESULTS: Glibenclamide (KATP channel blocker) increased perfusion pressure to a maximum fetoplacental arterial pressure of 37 +/- 6 mm Hg in controls versus 15 +/- 6 mm Hg in diabetes (P < .05). 4-Aminopyridine (KV channel blocker) equally increased fetoplacental arterial pressure in controls, and in diabetes (21 +/- 4 mm Hg vs 22 +/- 2 mm Hg). Apamin and charybdotoxin (KCa channel blockers) caused a negligible rise in fetoplacental arterial pressure. CONCLUSION: In the fetoplacental circulation, KATP channels and KV channels significantly contribute to baseline vascular tone. In diabetes, vascular KATP channel function is impaired. PMID- 15746701 TI - Ethical response to liability crisis. PMID- 15746702 TI - Risk factors for recurrent vulvovaginal candidiasis. PMID- 15746703 TI - A comparative review of the risks and benefits of hormone replacement therapy regimens. PMID- 15746704 TI - Research of surgical outcome of incontinence surgery in women. PMID- 15746705 TI - Effects of gastroschisis on gastric dilation. PMID- 15746706 TI - Cardiovascular death in patients with atrial fibrillation is better predicted by left atrial thrombus and spontaneous echocardiographic contrast as compared with clinical parameters. AB - We hypothesized that altered intra-atrial thrombogenicity, as reflected by the presence of left atrial (LA) thrombus or spontaneous echocardiographic contrast (SEC), would predict cardiovascular death in patients with atrial fibrillation (AF). In 175 patients with AF and no more than mild mitral regurgitation as detected by transesophageal echocardiography (TEE), 13 cardiovascular deaths occurred during a mean follow-up of 31 +/- 20 months. Multivariate logistic regression analysis using clinical variables identified the presence of congestive heart failure (relative risk [RR] = 4.22; P = .02) as the only positive predictor of cardiovascular death. However, when the TEE variables were added to the model, LA thrombus (RR = 5.52; P = .024) and LA SEC (RR = 7.96; P = .013) emerged as the only positive predictors of cardiovascular death. Kaplan Meier analysis demonstrated a lower event-free survival from cardiovascular death in patients with LA thrombus and/or SEC ( P = .0013). These findings support AF as a contributing cause of cardiovascular death independent of clinically associated risk factors, such as hypertension, diabetes mellitus, smoking, congestive heart failure, and prior myocardial infarction. PMID- 15746707 TI - Left atrial appendage assessment by second harmonic transthoracic echocardiography after an acute ischemic neurologic event. AB - Although second harmonic (sh) imaging is widely available in most contemporary ultrasound systems, its accuracy to evaluate left atrial appendage (LAA) morphology and function remains poorly characterized. We conducted a cross sectional survey of patients with acute ischemic neurologic conditions (n = 51) who underwent both transesophageal and transthoracic echocardiography (TTE) to explore the performance of sh in LAA assessment. Doppler and LAA area evaluation by sh TTE were feasible in most patients (98%). We observed positive and significant associations between sh TTE and transesophageal echocardiography assessment of LAA peak emptying velocities (r = 0.63, P < .001) and LAA maximum area (r = 0.73, P < .001). In addition, all patients (n = 7) with LAA thrombus or spontaneous contrast had peak emptying velocities less than 50 cm/s on sh TTE (negative predictive value of 100%). In multivariate analysis, LAA peak emptying velocity remained independently associated with LAA thrombus or contrast. In conclusion, sh TTE can provide valuable and clinically relevant information of LAA morphology and dynamics. PMID- 15746708 TI - Combined transesophageal echocardiography and transesophageal cardioversion probe: technical aspects. AB - A probe assembly for simultaneous transesophageal echocardiography and transesophageal cardioversion has been developed. This probe allows cardioversion with the delivery of much lower energy than the standard external approach. Details of the probe construction and its use are described, as is the prospect for future practice. The use of a combined probe may be the technique of choice for patients who require both cardioversion and transesophageal echocardiography. PMID- 15746709 TI - Cardiac resynchronization therapy acutely improves diastolic function. AB - BACKGROUND: Invasive studies have shown that cardiac resynchronization therapy (CRT) acutely improves left ventricular (LV) systolic performance and lowers filling pressures in a majority of patients with medically-refractory severe heart failure. Measurements included LV volume, ejection fraction, PWD early (E wave) and atrial (A-wave) velocities, diastolic filling time (DFT), and DTI early diastolic mitral annular velocity (Em) at the lateral and septal annulus; PWD mitral E-wave/Em and E/FP were calculated to estimate LV filling pressures. RESULTS: Immediately after CRT, LV volumes decreased and LVEF increased significantly. PWD mitral E-wave velocity decreased and E-wave duration and DFT increased significantly; mitral E/FP ratio also decreased significantly, consistent with a decrease in LV filling pressure. Patients with a pre-CRT mitral E/A ratio >1 (n = 20), demonstrated improvements in LV diastolic filling and lower filling pressures whereas those with an E/A ratio < or =1 (n = 21) did not show significant changes in diastolic indices. CONCLUSIONS: The acute effects of CRT include echocardiographic evidence of reduced LV volumes and increased LVEF with improved diastolic filling and lower filling pressures; LV relaxation is not significantly altered. The benefits in diastolic function are dependent on the PWD-determined LV filling characteristics prior to CRT. PMID- 15746710 TI - Isovolumic relaxation flow propagation velocity in patients with diseases impairing ventricular relaxation. AB - To evaluate the propagation velocity of isovolumic relaxation flow within the left ventricle (IRFPV) associated with impaired ventricular relaxation, 49 patients with diseases known to impair ventricular relaxation (disease group) and 38 age-matched control subjects (control group) were studied. IRFPV was measured as the slope of the first aliasing velocity line segment of the isovolumic relaxation flow wave front in the color M-mode Doppler echocardiogram. Compared with the control group, the disease group had thicker interventricular septum and left ventricular posterior wall, more left ventricular mass, and lower early diastolic mitral annular velocity (8 +/- 3 vs 11 +/- 4 cm/s, P < .001), early (E) wave propagation velocity (47 +/- 16 vs 70 +/- 41 cm/s, P = .002), and IRFPV (193 +/- 149 vs 395 +/- 220 cm/s, P < .001). No matter in subgroup or whole population analysis, either the early diastolic mitral annular velocity or the E wave propagation velocity was selected as one of the determinants of IRFPV. In conclusion, diseases impairing ventricular relaxation may retard IRFPV. PMID- 15746711 TI - Doppler tissue imaging improves assessment of abnormal interventricular septal and posterior wall motion in constrictive pericarditis. AB - We hypothesized that Doppler tissue imaging in the short axis would provide enhanced quantitative information for differentiating the pattern and extent of abnormal septal and posterior wall motion in constrictive pericarditis (CP). Using quantitative pulsed wave and color M-mode Doppler tissue imaging, we quantified the pattern of abnormal septal and posterior wall motion and studied its incremental advantage over conventional M-mode and 2-dimensional echocardiography in 40 patients with surgically proven CP. The pattern and extent of abnormalities were compared with 35 age- and sex-matched control subjects and 20 patients with abnormal septal motion of other causes. In 33 patients (82.5%) with CP, the interventricular septum showed high-velocity (>7 cm/s) early diastolic biphasic motion with or without multiple recoil waves (polyphasic diastolic septal fluttering). In the posterior wall, the early diastolic wave was normal but the late diastolic wave was reduced in 24 patients (60%) and absent in 7 (17.5%). In comparison, M-mode and 2-dimensional echocardiography identified abnormal septal or posterior wall motion in 24 patients (60%) ( P = .003). The pattern of abnormal septal motion in CP could be differentiated from abnormal septal motion of other causes in 16 patients (80%). The overall sensitivity and specificity of high-velocity polyphasic septal flutter for differentiating CP from control cases and other diseases was 82.5% and 92.7%, respectively. In CP, Doppler tissue imaging in the short axis provides unique diagnostic information and reliably differentiates CP from control cases and most other causes of abnormal septal motion. PMID- 15746712 TI - Double-orifice mitral valve with intact atrioventricular septum: an echocardiographic study with anatomic and functional considerations. AB - We identified 18 patients with double-orifice mitral valve (DOMV) and intact atrioventricular (AV) septum out of 40,179 echocardiographic studies performed between 1997 and 2002 at Children's Hospital, Denver, CO. In this study we describe (1) the anatomic characteristics of the DOMV in the absence of AV septal defect, (2) the function of the mitral valve by spectral and color Doppler flow mapping, and (3) associated lesions. The topographic location of the orifices in the leaflets suggests possible embryologic mechanisms of DOMV. In this series, DOMV was most commonly associated with left-sided obstructive lesions (in 39% of patients). Spectral and color Doppler interrogation demonstrated a normal flow profile in most cases; only 2 patients had significant mitral regurgitation or stenosis. Therefore, due to the uncertain natural history of this lesion and the potential need for endocarditis prophylaxis, careful imaging of the mitral valve is recommended, particularly in the presence of left-sided obstructive lesions. PMID- 15746713 TI - Anatomoechocardiographic correlation double inlet left ventricle. AB - Double inlet left ventricle (LV) is a type of atrioventricular connection in which the morphologically LV receives more than 50% of the atrioventricular valves when they are separate, or more than 75% of a common atrioventricular valve. The aim of this study was to establish an anatomoechocardiographic correlation between the morphologic features of equivalent anatomic specimens and the echocardiographic images of patients to provide a means of interpreting the image correctly and a more precise diagnosis of the cardiac defect. Echocardiography was used to study 18 patients with LV double inlet who were seen in a congenital heart disease clinic. The morphology of 17 hearts with this malformation from the department of embryology was analyzed to compare the anatomic features with their echocardiographic images. Echocardiography proved to be a noninvasive diagnostic tool that allowed characterization of anatomic and functional aspects of double inlet LV. PMID- 15746714 TI - A semiautomated ultrasound border detection program that facilitates clinical measurement of ultrasound carotid intima-media thickness. AB - We have developed a novel, semiautomated carotid intima-media thickness (CIMT) border detection program (AUTO) and evaluated its measurement reproducibility and accuracy. Images from 6 carotid segments were acquired in 50 subjects, for a total of 300 segments. Mean and maximum CIMT values were measured blindly at a reference (REF) lab and in duplicate by experienced (EXP) and novice (NOV) readers using manual (MAN) and AUTO methods. Coefficients of variation for AUTO measurements of mean (3.2%) and maximum (4.1%) CIMT were low, and the AUTO method improved the NOV reader's reproducibility. Compared with the REF lab, mean (0.012 +/- 0.006 mm) and maximum (0.144 +/- 0.006 mm) CIMT biases were small and equivalent to those of the REF lab ( P < .001). The AUTO method shortened reading times by 35% to 46% ( P < .001). We conclude that our novel AUTO CIMT measurement program improved reproducibility and was accurate. Compared with MAN tracing, the AUTO method agreed better with the REF lab and decreased reading time. PMID- 15746715 TI - Doppler echocardiography of 119 normal-functioning St Jude Medical mitral valve prostheses: a comprehensive assessment including time-velocity integral ratio and prosthesis performance index. AB - The purpose of this study was to provide, in a large number of patients, comprehensive Doppler echocardiographic assessment of normal St Jude Medical mitral valve prosthesis function using Doppler-derived hemodynamic variables, including the mitral valve prosthesis-to-left ventricular outflow tract time velocity integral ratio and prosthesis performance index. The pressure half-time was less than 130 milliseconds in all patients, and all but one patient had either a peak early mitral diastolic velocity of 2 m/s or less or a mitral valve prosthesis-to-left ventricular outflow tract time-velocity integral ratio of less than 2.2. There was a significant (P < .001) negative correlation between the prosthesis performance index and prosthesis size. This negative correlation suggests that there is more efficient use of the in vitro geometric orifice area with smaller prostheses. PMID- 15746716 TI - Use of hand-carried ultrasound devices to augment the accuracy of medical student bedside cardiac diagnoses. AB - BACKGROUND: Hand-carried ultrasound (HCU) devices used by cardiologists as extensions of the physical examination have been shown to improve the accuracy of bedside diagnoses. We tested the feasibility of teaching medical students to use HCU devices to make bedside cardiac diagnoses and compared the accuracy of their HCU and physical examinations. METHODS: In all, 10 fourth-year medical students enrolled in a 4-week medical school course on the cardiac examination. Students examined 12 standardized patients at 3 different time intervals: (1) on day 1 of the course; (2) on day 10 after review of cardiac physical examination using traditional teaching methods; and (3) after instruction on the use of HCU devices. Students were scored at each time interval for primary findings (most salient) and all findings, accounting for both errors of commission and omission. Scores could range from +12 to -12 for primary findings and from +22 to -22 for all findings. A perfect score was +12 for primary findings and +22 for all findings. RESULTS: The average score for all students at baseline was -3.2 +/- 3.1 and -5.7 +/- 4.8 for primary and all findings, respectively. A significant improvement in the scores was noted with use of the HCU device (2.6 +/- 3.1 and 5.2 +/- 6.6 for primary and all findings, respectively) compared with the baseline and two subsequent physical examinations. CONCLUSION: Instruction of fourth-year medical students on the use of HCU device is feasible and results in significantly more accurate bedside diagnoses. PMID- 15746717 TI - The research grant process: a reference guide for cardiovascular sonographers. AB - Opportunities for sonographer-initiated research have increased during the past decade. Although research has traditionally been viewed as a nonclinical activity, funding is available for clinical problems. This article aims to increase sonographer awareness of this unique opportunity, encourage the submission of grant proposals, and enhance the understanding of the grant writing process. This review discusses the procedures involved in planning a research project while describing the structure of the research grant proposal including specific aims, background, preliminary studies, methods, potential limitations, significance, budget, and references. PMID- 15746718 TI - Infundibular ventricular septal defect, aneurysm of the sinus of Valsalva, and bicuspid aortic valve in a caucasian male. AB - We report the case of a Caucasian man with the unusual combination of an infundibular ventricular septal defect (VSD), an aneurysm of the sinus of Valsalva communicating with the right ventricle, and a bicuspid aortic valve. When aortic valve regurgitation associated with mild right aortic cusp prolapse appeared during follow-up examination, the patient, although totally asymptomatic, underwent surgical repair of the aortic sinus and closure of the VSD. PMID- 15746719 TI - Ventricular septal hamartoma mimicking hypertrophic cardiomyopathy in a 41-year old woman presenting with paroxysmal supraventricular tachycardia. AB - We report an unusual case of cardiac hamartoma involving the ventricular septum. The patient presented with palpitations and exertional chest pain. Echocardiography indicated a large homogeneous mass occupying the superior two thirds of the ventricular septum. This report confirms the use of transthoracic echocardiography as a screening tool in the diagnosis and management of an uncommon case of a benign cardiac tumor. PMID- 15746720 TI - Calcified right ventricular mass and pulmonary embolism in a previously healthy young woman. AB - A 20-year-old woman with a recent episode of pulmonary embolism was found by echocardiographic examination to have a calcified apical right ventricular intracavitary mass. She had a history of cigarette smoking and had been taking oral contraceptive pills. At age 17 years, she had sustained substantial blunt chest injury during a motor vehicle accident. The intracardiac mass was resected and represented an old calcified mural thrombus. It is postulated that its development was related to cardiac trauma. These intracavitary masses have been referred to as cardiac calcified amorphous tumors and generally represent old organized and focally calcified mural thrombus. They should be distinguished from calcified benign or malignant neoplasms of the heart. PMID- 15746721 TI - Transesophageal echocardiographic image of a thrombus crossing a persistent foramen ovale from the right into the left atrium. PMID- 15746722 TI - Superior vena cava (SVC) thrombosis: diagnosis and follow up by transesophageal echocardiography (TEE). AB - Hemodialysis catheters (HC) were developed as a vascular access, intended to be a bridging device until a permanent access is placed. Although many reports exist of indwelling catheter-related thrombosis, optimal diagnostic study and therapeutic options are yet to be established. We are presenting a case with HC thrombosis diagnosed by TEE managed conservatively with excellent long-term outcome. PMID- 15746723 TI - The abdominal aorta and constrictive pericarditis: abdominal aortic respiratory variation as an echocardiographic finding in constrictive pericarditis. PMID- 15746724 TI - Myofibroblastic sarcoma of mitral valve: a case report. AB - We report a patient with symptoms of congestive heart failure whose workup revealed a myofibroblastic sarcoma of mitral valve. PMID- 15746725 TI - Guidelines and recommendations for digital echocardiography. PMID- 15746726 TI - Influence of preload alterations on parameters of systolic left ventricular long axis function: a Doppler tissue study: flawed methodology leads to spurious results. PMID- 15746727 TI - Presidential address: an evolving discipline. PMID- 15746728 TI - Endoscopic saphenous vein harvesting: the good, the bad, and the ugly. PMID- 15746729 TI - The Fontan operation, thromboembolism, and anticoagulation: a reappraisal of the single bullet theory. PMID- 15746730 TI - Randomized trial of endoscopic versus open vein harvest for coronary artery bypass grafting: six-month patency rates. AB - OBJECTIVE: We sought to compare the 6-month angiographic patency rates of greater saphenous veins removed during coronary artery bypass grafting with the endoscopic vein harvest or open vein harvest techniques. METHODS: Two hundred patients undergoing nonemergency on-pump coronary artery bypass grafting were prospectively randomized to either endoscopic vein harvest or open vein harvest. Follow-up angiography of all vein grafts was scheduled at 6 months. Graft patency and disease grades were assigned independently by 2 interventional cardiologists. Leg wound healing was evaluated at discharge, 1 month, and 6 months for evidence of complications. RESULTS: There were 3 conversions from endoscopic vein harvest to open vein harvest because of vein factors. Leg wound complications were significantly lower in the endoscopic vein harvest group (7.4% vs 19.4%, P = .014). On multivariable analysis, endoscopic vein harvest emerged as the only factor affecting wound complications (odds ratio, 0.33). Three deaths (2 perioperative and 1 late) occurred in the endoscopic vein harvest group that were unrelated to vein graft closure. Twenty-four and 29 patients in the endoscopic vein harvest and open vein harvest cohorts, respectively, refused the follow-up 6 month angiography. Therefore a total of 144 angiograms (73 endoscopic vein harvests and 71 open vein harvests) and 336 vein grafts (166 endoscopic vein harvests and 170 open vein harvests) were available for analysis. The overall occlusion rates at 6 months were 21.7% for endoscopic vein harvest and 17.6% for open vein harvest. Additionally, there was evidence of significant disease (>50% stenosis) in 10.2% and 12.4% of endoscopic vein harvest and open vein harvest grafts, respectively. By means of ordinal hierarchic logistic regression, endoscopic vein harvest was not found to be a risk factor for vein graft occlusion or disease (odds ratio, 1.15). Significant predictors were congestive heart failure (odds ratio, 2.87), graft to the diagonal artery territory (odds ratio, 1.76), larger vein conduit size (odds ratio, 1.32), and graft flow (odds ratio, 0.90). CONCLUSION: Endoscopic vein harvest reduces leg wound complications compared with open vein harvest without compromising the 6-month patency rate. The overall patency rate depends on target and vein-related variables and patient characteristics rather than the method of vein harvesting. PMID- 15746731 TI - Surgical treatment of ischemic mitral regurgitation might not influence ventricular remodeling. AB - OBJECTIVES: Surgical treatment for ischemic mitral regurgitation has become more aggressive. However, no clinical study has demonstrated that surgical correction of chronic ischemic mitral regurgitation improves survival. We used 4 well developed ovine models of postinfarction left ventricular remodeling to test the hypothesis that ischemic mitral regurgitation does not significantly contribute to postinfarction left ventricular remodeling. METHODS: Infarction of 21% to 24% of the left ventricular mass was induced by means of coronary ligation in 77 sheep. Infarctions varied only by anatomic location in the left ventricle: anteroapical, n = 26; anterobasal, n = 16; laterobasal, n = 9; and posterobasal, n = 20. Six additional sheep had ring annuloplasty before posterobasal infarction. End-systolic and end-diastolic left ventricular volume, end-systolic muscle-to-cavity area ratio, left ventricular sphericity, ejection fraction, and degree of ischemic mitral regurgitation, as determined by means of quantitative echocardiography, were assessed before infarction and at 2, 5, and 8 weeks after infarction. RESULTS: All infarcts resulted in significant postinfarction remodeling and decreased ejection fraction. Anteroapical infarcts lead to left ventricular aneurysms. Only posterobasal infarcts caused severe and progressive ischemic mitral regurgitation. Remodeling because of posterobasal infarcts was not more severe than that caused by infarcts at other locations. Furthermore, prophylactic annuloplasty prevented the development of mitral regurgitation after posterobasal infarction but had no effect on remodeling. CONCLUSION: The extent of postinfarction remodeling is determined on the basis of infarct size and location. The development of ischemic mitral regurgitation might not contribute significantly to adverse remodeling. Ischemic mitral regurgitation is likely a manifestation rather than an important impetus for postinfarction remodeling. PMID- 15746732 TI - Left ventricular mass regression after aortic valve replacement with 17-mm St Jude Medical mechanical prostheses in isolated aortic stenosis. AB - OBJECTIVE: The present study investigated the outcomes of aortic valve replacement with 17-mm mechanical prostheses in patients with isolated aortic stenosis. METHODS: Between January 1997 and January 2003, 35 patients (mean age, 63.4 +/- 17 years; median age, 70 years; age range, 16-84 years) underwent isolated aortic valve replacement with a 17-mm St Jude Medical Hemodynamic Plus (16 [45.7%] patients) or a St Jude Medical Regent prosthesis (19 [54.3%] patients). The paired Student t test or the paired Wilcoxon rank sum test were used to compare preoperative with follow-up echocardiographic measurements. RESULTS: Thirty-two (91.4%) patients were female, mean height was 154.4 +/- 8.3 cm, mean weight was 62.2 +/- 9.2 kg, and mean body surface area was 1.59 +/- 0.13 m 2 . The preoperative average New York Heart Association class was 2.8 +/- 0.8. The mean preoperative left ventricular mass index was 135.2 +/- 31 g/m 2 . Preoperative echocardiography showed an average gradient of 65.7 +/- 19.2 mm Hg (mean) and 103.6 +/- 30.7 mm Hg (peak) and a mean indexed effective orifice area of 0.40 +/- 0.1 cm 2 /m 2 . Echocardiographic follow-up time averaged 28.2 +/- 22.7 months (range, 13-72 months). Follow-up was 100% complete (1131.7 patient months). Hospital mortality was 8.6% (3 patients). Actuarial 5-year survival was 94.7%. The mean postoperative New York Heart Association class was 1.13 +/- 0.34 ( P < .001), with 27 (87.1%) patients in class I and 4 patients in class II. A significant regression of the indexed left ventricular mass was found (postoperative mean value, 107.8 +/- 22.8 g/m 2 ; P < .0001), despite a mean indexed effective orifice area of 0.67 +/- 0.14 cm 2 /m 2 (median, 0.66 cm 2 /m 2 ). CONCLUSIONS: Selected patients with aortic stenosis can experience satisfactory clinical improvement and significant indexed left ventricular mass regression after aortic valve replacement with modern small-diameter bileaflet prostheses. PMID- 15746733 TI - Acute poststernotomy mediastinitis managed with debridement and closed-drainage aspiration: factors associated with death in the intensive care unit. AB - OBJECTIVE: The purpose of the study is to describe an intensive care unit's experience in the treatment of poststernotomy mediastinitis and to identify factors associated with intensive care unit death. METHODS: Over a 10-year period, 316 consecutive patients with mediastinitis occurring less than 30 days after sternotomy were treated in a single unit. First-line therapy was closed drainage aspiration with Redon catheters. Variables recorded, including patient demographics, underlying disease classification, clinical and biologic data available at intensive care unit admission and day 3, and their association with intensive care unit mortality, were subjected to multivariate analyses. RESULTS: Intensive care unit mortality (20.3%) was significantly associated with 5 variables available at admission: age greater than 70 years (odds ratio, 2.70), operation other than coronary artery bypass grafting alone (odds ratio, 2.59), McCabe class 2/3 (odds ratio, 2.47), APACHE II score (odds ratio, 1.12 per point), and organ failure (odds ratio, 2.07). After introducing day 3 variables into the logistic regression model, independent risk factors for intensive care unit death were as follows: age greater than 70 years, operations other than coronary artery bypass grafting alone, McCabe class 2/3, APACHE II score, mechanical ventilation still required on day 3, and persistently positive bacteremia. For patients receiving mechanical ventilation for less than 3 days, mortality was very low (2.4%). In contrast, for patients receiving mechanical ventilation for 3 days or longer, mortality reached 52.8% and was associated with non-coronary artery bypass grafting cardiac surgery, persistently positive bacteremia, and underlying disease. CONCLUSIONS: In patients requiring intensive care for acute poststernotomy mediastinitis, age, type of cardiac surgery, underlying disease, and severity of illness at the time of intensive care unit admission were associated with intensive care unit death. Two additional factors (mechanical ventilation dependence and persistently positive bacteremia) were identified when the analyses were repeated with inclusion of day 3 patient characteristics. PMID- 15746734 TI - Imbalanced chordal force distribution causes acute ischemic mitral regurgitation: mechanistic insights from chordae tendineae force measurements in pigs. AB - BACKGROUND: Ischemic mitral regurgitation is caused by an imbalance of the entire mitral-ventricular complex. This interaction is mediated through the chordae tendineae force distribution, which may perturb several elements of the mitral valve apparatus. Our objective was to investigate the association between the mitral valvular 3-dimensional geometric perturbations and chordae tendineae force redistribution in a porcine model of acute ischemic mitral regurgitation. METHODS: In 9 pigs, acute ischemic mitral regurgitation was induced by repeated microembolization of the left circumflex coronary artery. Mitral leaflet coaptation geometry was determined by 2-dimensional echocardiography and reconstructed 3-dimensionally. Leading edge chordal forces were measured by dedicated miniature force transducers at control and during ischemic mitral regurgitation. RESULTS: During acute ischemic mitral regurgitation, there was a decreased tension of the primary chorda from the ischemic posterior left ventricular wall to the anterior leaflet (0.295 +/- 0.063 N vs 0.336 +/- 0.071 N [control]; P < .05). The tension of the chorda from the nonischemic anterior left ventricular wall to the anterior leaflet increased (0.375 +/- 0.066 N vs 0.333 +/ 0.071 N [control]; P < .05). In accordance, relative leaflet prolapse was observed at the ischemic commissural side, whereas there was an increase in the leaflet surface area at the nonischemic commissural side, indicating localized leaflet tethering. CONCLUSIONS: Acute ischemic mitral regurgitation due to posterior left ventricular wall ischemia was associated with focal chordal and leaflet tethering at the nonischemic commissural portion of the mitral valve and a paradoxical decrease of the chordal forces and relative prolapse at the ischemic site of the anterior mitral valve leaflet. PMID- 15746735 TI - Is postoperative calcium channel blocker therapy needed in patients with radial artery grafts? AB - BACKGROUND: Chronic calcium channel blocker therapy has traditionally been considered necessary in patients carrying a radial artery graft, even in the absence of objective data to support it. This report was conceived to evaluate the angiographic and clinical effects of calcium channel blocker therapy during the first postoperative year. PATIENTS AND RESULTS: A total of 100 consecutive patients who received a radial artery graft at our institution were randomly assigned to receive (n = 53) or not receive (n = 47) calcium channel blocker therapy with oral diltiazem 120 mg/daily started in the early postoperative period. At 1-year follow-up, all patients were reassessed clinically and by Tl 201 myocardial scintigraphy, and 83 of them underwent control angiography. In 12 cases we also evaluated the response of the radial artery to the endovascular infusion of serotonin. No difference in terms of clinical outcome, scintigraphic results, and patency rate was found between patients who received or did not receive calcium channel blocker therapy. Endovascular serotonin infusion evoked an evident spastic reaction of radial artery grafts, not attenuated by calcium channel blocker therapy. CONCLUSION: Calcium channel blocker therapy started immediately after surgery and continued for the first postoperative year does not affect radial artery graft patency and clinical and scintigraphic outcomes. On the basis of these data, the prophylactic use of calcium channel blocker therapy in patients with radial artery grafts seems unsubstantiated. PMID- 15746736 TI - The effect of bilateral internal thoracic artery harvesting on superficial and deep sternal infection: The role of skeletonization. AB - OBJECTIVE: To determine the relative risk of sternal dehiscence in patients undergoing bilateral internal thoracic artery harvesting and to assess whether and to what extent the technique of artery skeletonization might reduce this risk. METHODS: Prospectively collected data on patients undergoing coronary artery bypass operations with at least a single internal thoracic artery were reviewed. The last 450 patients receiving bilateral internal thoracic artery grafts were compared with 450 patients who received a single internal thoracic artery during the same period. The left internal thoracic artery was always harvested in a pedicled fashion. Among patients receiving a bilateral internal thoracic artery, both arteries were harvested in a pedicled fashion in 300 cases, whereas both internal thoracic arteries were skeletonized in the remaining 150 cases. RESULTS: Compared with a single internal thoracic artery, harvesting both internal thoracic arteries either in a skeletonized or in a pedicled fashion increased the chance of deep (1.1% vs 3.3% vs 4.7%; P = .01) or superficial (4.8% vs 7.8% vs 12%; P = .002) sternal infection. However, the technique of artery harvesting (odds ratio, 4.1; 95% confidence interval, 1.4-12.1); the presence of peripheral arteriopathy (odds ratio, 3.1; 95% confidence interval, 1.2-8.5), and resternotomy for bleeding (odds ratio, 8.2; 95% confidence interval, 2.0-33.6) were the only independent predictors for deep sternal infection, whereas the technique of artery harvesting (odds ratio, 3.0; 95% confidence interval, 1.6 5.4), female sex (odds ratio, 2.2; 95% confidence interval, 1.2-4.2), and diabetes (odds ratio, 1.7; 95% confidence interval, 1.0-2.9) were the only independent predictors of superficial sternal infection. In diabetic patients, there was no difference in the incidence of deep sternal infection among patients receiving a single internal thoracic artery or double skeletonized internal thoracic arteries ( P = .4). CONCLUSIONS: Bilateral internal thoracic artery harvesting carries a higher risk of sternal infection than harvesting a single internal thoracic artery. Skeletonization of both internal thoracic arteries significantly decreases this risk. A strategy of bilateral thoracic artery grafting can also be offered to patients at high risk for wound infection. PMID- 15746737 TI - Efficacy of intraluminal pulmonary artery banding. AB - OBJECTIVE: The extraluminal technique of pulmonary artery banding can be difficult to perform precisely in conjunction with cardiopulmonary bypass and is associated with a significant risk of band-related complications. We analyzed our results with an intraluminal technique of pulmonary artery banding in patients who required cardiopulmonary bypass for the performance of associated cardiac repairs. METHODS: The medical records of 18 neonates and infants who underwent intraluminal pulmonary artery banding were retrospectively reviewed. A circular patch with a 3.0-mm, 3.6-mm, or 4.0-mm diameter fenestration was sutured to the inner circumference of the main pulmonary artery. Preoperative, intraoperative, and postoperative variables were reviewed to assess the efficacy and safety of the intraluminal technique. RESULTS: Intraluminal pulmonary artery banding produced a consistent and significant reduction in the systolic pulmonary artery pressure (64.00 +/- 12.24 to 16.53 +/- 6.33 mm Hg, P < .001), the systolic pulmonary artery pressure/systolic systemic pressure ratio (0.91 +/- 0.10 to 0.19 +/- 0.07, P < .001), and the pulmonary flow/systemic flow ratio (4.32 +/- 3.04 to 0.91 +/- 0.49, P = .015). There were no band-related anatomic complications. Two patients did require percutaneous dilation of the intraluminal pulmonary artery band before debanding to palliate systemic arterial desaturation. CONCLUSIONS: Intraluminal pulmonary artery banding is an effective palliative procedure that can be used in patients who require cardiopulmonary bypass for the performance of cardiac repairs in addition to placement of the pulmonary artery band. PMID- 15746738 TI - Complex aortic valve repair as a durable and effective alternative to valve replacement in children with aortic valve disease. AB - OBJECTIVE: This study was undertaken to determine the utility of aortic valve repair in children. METHODS: A retrospective analysis was conducted on aortic valve surgery from 1973 to 2004 at Children's Hospital of Wisconsin. RESULTS: Procedures were classified as simple repairs (blunt valvotomy, commissurotomy with or without thinning, n = 147), repair of aortic insufficiency with ventricular septal defect (n = 22), complex repairs (any combination of additional procedures including suspension of prolapsed leaflets, leaflet extensions, repair of torn or perforated leaflets, annuloplasty, reduction of sinus of Valsalva plasty, and concomitant repair of supravalvular or subvalvular stenosis, n = 57), and replacements (n = 57, 20 mechanical, 2 porcine, and 35 human valves). Freedoms from reintervention for simple repairs and repair of aortic insufficiency with ventricular septal defect at 10 years were 86% +/- 5% and 93.3% +/- 6%, respectively. For complex valve repair, freedoms from reintervention at 1, 5, and 10 years were 94% +/- 3%, 85% +/- 6%, and 44% +/- 15%, versus 96% +/- 3%, 77% +/- 9%, and 77% +/- 9% for valve replacement ( P = .3). At intermediate follow-up, patients with complex valve repair had a residual gradient of 20 +/- 21 mm Hg, and 94% were free of severe aortic insufficiency. Residual aortic stenosis ( P < .05) but not the preoperative diagnosis of combined aortic stenosis and insufficiency predicted the need for reintervention. CONCLUSION: Freedom from reintervention after complex valve repairs was not different from that after valve replacement, with acceptable residual aortic stenosis and insufficiency. Simple repairs and repair of aortic insufficiency with ventricular septal defect yielded excellent long-term freedom from reintervention. PMID- 15746739 TI - The effect of repair technique on postoperative right-sided obstruction in patients with truncus arteriosus. AB - OBJECTIVES: We reviewed our experience with repair of truncus arteriosus to assess the effect of type of right ventricular outflow tract reconstruction on perioperative morbidity, survival, and freedom from catheter-based interventions and reoperation. METHODS: Patients undergoing repair of truncus arteriosus from June 1990 through February 2004 were evaluated on the basis of operative procedure regarding preoperative and postoperative variables, the need for postoperative catheter-based intervention or reoperation, and survival on the basis of univariate, multivariable, and actuarial analyses. RESULTS: Of 54 study patients, 15 (28%) received a valved homograft, and 39 (72%) received a direct connection with a variety of hood materials. Five (9.1%) patients died. Valved homograft recipients were more likely to require reoperation than patients receiving direct connections (40% vs 15%, P = .046); however, valved homograft and direct connection recipients had a similar incidence of the combined end point of reoperation or catheter-based intervention (40.0% vs 37.5%, P = .865). Univariate and multivariable modeling demonstrated use of valved homografts or direct connections with an autologous pericardial hood to be predictive of the need for later catheter-based intervention or reoperation. Actuarial analysis demonstrated a trend toward improved outcomes in the direct connection group and within the direct connection cohort, a statistically significant difference on the basis of hood type. CONCLUSIONS: Although the direct connection technique might not prevent later catheter-based intervention, it does reduce the need for reoperation. Outcomes among direct connection recipients were associated with hood type: polytetrafluoroethylene hoods (W. L. Gore & Associates, Inc, Tempe, Ariz) had the lowest rate of reintervention, and untreated autologous pericardial hoods had the highest rate of reintervention. We report excellent outcomes with primary repair of truncus arteriosus. Where anatomically appropriate, we advocate the direct connection technique. PMID- 15746740 TI - Prophylaxis of thromboembolic complications after the Fontan operation (total cavopulmonary anastomosis). AB - OBJECTIVES: Thrombotic events have been reported as a major cause of morbidity after the Fontan procedure. There is no consensus concerning the postoperative mode and duration of anticoagulation prophylaxis. In a retrospective study, we evaluated the results of a prophylactic regimen on the basis of the surgical technique, potentially predisposing risk factors, and specific sequelae. METHODS: We evaluated 142 surviving patients after total cavopulmonary anastomosis (mean follow-up was 91.1 +/- 43.9 months). Prophylactic antithrombotic treatment was initiated in 86 patients with partial prosthetic venous pathway with acetylsalicylic acid; 45 patients with complete autologous tissue venous pathway or partial prosthetic venous pathway received no anticoagulation, and 11 patients received warfarin sodium (Coumadin). During long-term follow-up, 22 patients (12 after acetylsalicylic acid medication) crossed over to warfarin. RESULTS: Thrombotic events occurred in 10 patients (7%), with systemic venous thrombus formation in 8 (5.6%), stroke in 2 (1.4%), and a peak incidence during the first postoperative year. Eight of 10 patients were receiving heparin therapy mainly for prolonged postoperative immobilization. During follow-up, none of the 74 patients receiving acetylsalicylic acid and 1 of 40 patients without medication presented with thrombus formation. Under warfarin medication, 1 of 28 patients had an asymptomatic thrombus. Expected freedom from a thromboembolic event was 92% at 5 years and 79% at 10 years. There was no association with coagulation factor abnormalities. Protein-losing enteropathy was present in 4 of 10 patients. CONCLUSION: A prophylactic anticoagulation strategy that considers the surgical technique and potential predisposing circumstances proved effective in the prevention of late thrombotic complications after total cavopulmonary anastomosis. There is no need for routine anticoagulation during long-term follow up after Fontan-type surgery in pediatric patients. PMID- 15746741 TI - Risk factors for cognitive dysfunction after coronary artery bypass graft surgery in patients with type 2 diabetes. AB - OBJECTIVES: The mechanisms of postoperative cognitive dysfunction in patients with diabetes after coronary artery bypass grafting are not fully understood. We sought to determine which type 2 diabetes-related factors contributed to postoperative cognitive dysfunction at 7 days and 6 months after coronary artery bypass grafting. METHODS: One hundred eighty patients with type 2 diabetes who were scheduled for elective coronary artery bypass grafting were studied. As a control group, 100 patients without diabetes mellitus matched for age, sex, and educational level were examined. Hemodynamic parameters (arterial and jugular venous blood gas values) were measured during cardiopulmonary bypass. All patients underwent a battery of neurologic and neuropsychologic tests the day before surgery, 7 days after surgery, and 6 months after surgery. RESULTS: Age (odds ratio 1.5, 95% confidence interval 1.3-1.8, P = .03), presence of hypertension (odds ratio 1.8, 95% confidence interval 1.3-2.0, P = .01), jugular venous oxygen saturation less than 50% time (odds ratio 1.5, 95% confidence interval 1.1-2.0, P = .045), presence of ascending aorta atherosclerosis (odds ratio 1.5, 95% confidence interval 1.1-2.6, P = .01), diabetic retinopathy (odds ratio 2.0, 95% confidence interval 1.3-3.0, P = .01), and insulin therapy (odds ratio 2.0, 95% confidence interval 1.3-3.0, P = .05), were associated with cognitive impairment at 7 days. Insulin therapy (odds ratio 2.0, 95% confidence interval 1.3-3.8, P = .01), diabetic retinopathy (odds ratio 1.3, 95% confidence interval 1.2-2.9, P < .01), and hemoglobin A 1c (odds ratio 1.9, 95% confidence interval 1.3-3.1, P = .047) were associated with cognitive impairment at 6 postoperative months. CONCLUSIONS: Insulin therapy, diabetic retinopathy, and hemoglobin A 1c were factors in cognitive impairment at 7 days and 6 months after coronary artery bypass grafting in patients with type 2 diabetes. PMID- 15746742 TI - Dynamic and differential changes in myocardial and plasma endothelin in patients undergoing cardiopulmonary bypass. AB - OBJECTIVE: The bioactive peptide endothelin modulates left ventricular function by changing afterload, coronary vascular tone, and myocardial contractility. However, whether increased plasma endothelin levels observed in patients during and after coronary revascularization and cardiopulmonary bypass reflect actual myocardial interstitial levels are unknown. METHODS: A microdialysis probe (outer diameter: 0.77 mm; length: 4 mm) was placed in the left ventricular apical midmyocardium in 20 patients and myocardial interstitial fluid was collected (2.5 microL/min) at baseline and up to 30 minutes after cardiopulmonary bypass. Myocardial interstitial and systemic arterial endothelin were measured by radioimmunoassay. RESULTS: Baseline myocardial interstitial endothelin was over 6 fold higher than plasma (20.11 +/- 2.07 vs 3.19 +/- 0.25 fmol/mL, P < .05). Plasma endothelin increased by 23% +/- 12% at 60 minutes of cardiopulmonary bypass whereas myocardial interstitial endothelin increased by 105% +/- 24%, P < .05), and this change was higher than in the plasma ( P < .05). Although no further change in plasma endothelin occurred during cardiopulmonary bypass, myocardial interstitial levels increased further after crossclamp removal (400% +/- 75%) and remained significantly higher than plasma at separation from cardiopulmonary bypass. CONCLUSION: The unique findings of this study were 2 fold: First, significant compartmentalization of endothelin exists within the human myocardium. Second, a significantly higher and temporally disparate change in myocardial interstitial endothelin occurs during and after cardiopulmonary bypass when compared with systemic levels. These dynamic changes in myocardial endothelin likely influence coronary vascular tone and contractility. PMID- 15746743 TI - Side differences in cerebrovascular accidents after cardiac surgery: a statistical analysis of neurologic symptoms and possible implications for anatomic mechanisms of aortic particle embolization. AB - BACKGROUND: Aortic manipulation and particle embolization have been identified to cause cerebrovascular accidents in cardiac surgery. Recent data suggest that left hemispheric cerebrovascular accident (right-sided symptoms) is more common, and this has been interpreted as being caused by aortic cannula stream jets. Our aim was to evaluate symptoms of cerebrovascular accident and side differences from a retrospective statistical analysis. METHODS: During a 2-year period, 2641 consecutive cardiac surgery cases were analyzed. Patients positive for cerebrovascular accident were extracted from a database designed to monitor clinical symptoms. A protocol was used to confirm symptom data with the correct diagnosis in patient records. Patients were subdivided into 3 groups: control, immediate cerebrovascular accident, and delayed cerebrovascular accident. RESULTS: Among pooled patients, immediate and delayed cerebrovascular accidents were 3.0% and 0.9%, respectively. The expected predisposing factors behind immediate cerebrovascular accidents were significant, although the type of operation affected this search. Aortic quality was a strong predictor ( P < .001). The rate of delayed cerebrovascular accident was unaffected by surgery group. Left-sided symptoms of immediate cerebrovascular accident were approximately twice as frequent ( P = .016) as on the contralateral side. This phenomenon was observed for pooled patients and for isolated coronary bypass procedures (n = 1882; P = .025). CONCLUSIONS: Immediate cerebrovascular accident and aortic calcifications are linked. The predominance of left-sided symptoms may suggest that aortic manipulation and anatomic mechanisms in the aortic arch are more likely to cause cerebrovascular accidents than effects from cannula stream jets. PMID- 15746744 TI - Polarized arrest with warm or cold adenosine/lidocaine blood cardioplegia is equivalent to hypothermic potassium blood cardioplegia. AB - BACKGROUND: Hypothermic depolarizing hyperkalemic (K + 20 mEq/L) blood cardioplegia is the "gold standard" in cardiac surgery. K + has been associated with deleterious consequences, eg, intracellular calcium overload. This study tested the hypothesis that elective arrest in a polarized state with adenosine (400 micromol/L via adenosine triphosphate-sensitive potassium channel opening) and the Na + channel blocker lidocaine (750 micromol/L) as the arresting agents in blood cardioplegia provides cardioprotection comparable to standard hypothermic K + -blood cardioplegia. METHODS: Anesthetized dogs were placed on cardiopulmonary bypass and assigned to 1 of 3 groups receiving antegrade cardioplegia delivered every 20 minutes for 1 hour of arrest: cold (10 degrees C) K + -blood cardioplegia (n = 6), cold (10 degrees C) adenosine/lidocaine blood cardioplegia (n = 6), or warm (37 degrees C) adenosine/lidocaine blood cardioplegia (n = 6). After an hour of arrest, cardiopulmonary bypass was discontinued, and reperfusion was continued for 120 minutes. RESULTS: Time to arrest was longer with cold and warm adenosine/lidocaine blood cardioplegia (175 +/- 19 seconds and 143 +/- 19 seconds, respectively) compared with K + -blood cardioplegia (27 +/- 2 seconds; P < .001). Postcardioplegia left ventricular systolic function (slope of the end-systolic pressure/dimension relationship) was comparable among the 3 groups (K + -blood cardioplegia, 15.2 +/- 2.1 mm Hg/mm; cold adenosine/lidocaine blood cardioplegia, 15.9 +/- 3.4 mm Hg/mm; warm adenosine/lidocaine blood cardioplegia, 14.1 +/- 2.8 mm Hg/mm; P = .90). Plasma creatine kinase activity in cold and warm adenosine/lidocaine blood cardioplegia was similar to that in K + -blood cardioplegia at 120 minutes of reperfusion (cold adenosine/lidocaine blood cardioplegia, 11.5 +/- 2.1 IU/g protein; warm adenosine/lidocaine blood cardioplegia, 10.1 +/- 0.9 IU/g protein; K + -blood cardioplegia, 7.6 +/- 0.8 IU/g protein; P = .17). Postcardioplegia coronary artery endothelial function was preserved in all groups. CONCLUSIONS: Intermittent polarized arrest with warm or cold adenosine/lidocaine blood cardioplegia provided the same degree of myocardial protection as intermittent hypothermic K + -blood cardioplegia in normal hearts. PMID- 15746745 TI - Assessment of minimally invasive direct coronary artery bypass grafting of the left internal thoracic artery by means of magnetic resonance imaging. AB - OBJECTIVES: We sought to evaluate graft patency, flow, and flow reserve in patients with minimally invasive direct coronary artery bypass surgery of internal thoracic artery grafts by a combined magnetic resonance protocol with a phase-contrast technique and magnetic resonance angiography. METHODS: At 1.5 T (Magnetom Sonata, Siemens), 30 symptomatic patients with 30 left internal thoracic artery grafts were examined 6 years after minimally invasive surgical intervention. Navigator-gated magnetic resonance angiography and contrast enhanced FLASH-3D magnetic resonance angiography (0.2 mmol gadopentate-diethylene triamine pentetic acid [Gd-DTPA]/kg body weight) was used to assess bypass patency. Phase-contrast flow measurements with retrospective gating were performed in the internal thoracic artery grafts at rest and after stress induction with dipyridamole (0.57 mg/kg body weight). Graft patency was evaluated by means of multidetector computed tomography (Sensation 16, Siemens). RESULTS: Internal thoracic artery grafts were occluded in 5 of 30 patients. In 6 patients the anastomosis to the left anterior descending artery was highly stenotic (>70 % ) at multidetector computed tomography. In patients with regular grafts (multidetector computed tomography), a significant improvement of graft flow ( P < .001) and diastolic/systolic peak velocity ratio ( P < .001) after stress induction was detected. Magnetic resonance angiography combined with flow reserve measurements could differentiate between occluded-stenotic and regular minimally invasive direct coronary artery bypass grafts. CONCLUSIONS: Magnetic resonance imaging allows a combined assessment of bypass patency and flow with flow reserve in patients after the minimally invasive direct coronary artery bypass operation. The protocol of this study might be applicable for the evaluation of graft status in symptomatic patients after revascularization. PMID- 15746746 TI - Two-dimensional and 3-dimensional optical coherence tomographic imaging of the airway, lung, and pleura. AB - BACKGROUND: Methods for obtaining real-time in vivo histologic resolution by means of noninvasive endoscopic optical imaging would be a major advance for thoracic surgical diagnostics and treatment. Optical coherence tomography is a rapidly evolving technology based on near-infrared interferometry that might provide these capabilities. The purpose of this study is to investigate the feasibility of real-time 2- and 3-dimensional optical coherence tomographic imaging of airway, pleural, and subpleural lung tissues in normal, inflammatory, and malignant animal models and patients with known or suspected airway malignancy. METHODS: Freshly excised lungs and pleural tissue obtained from rabbits with inhalation lung injury and induced empyema, metastatic sarcomas, and pleural sarcomas and from patients with airway disease were imaged by using 2- and 3-dimensional optical coherence tomography with a prototype superluminescent diode optical coherence tomographic system constructed in our laboratory. Lungs and pleural tissue were subsequently processed for standard hematoxylin and eosin histology for comparison with optical coherence tomography. RESULTS: Optical coherence tomographic imaging achieved an ex vivo resolution of 10 microm and an in vivo resolution of about 30 microm with a depth penetration of 1 to 2 mm with 2- and 3- dimensional reconstruction capabilities. Tumors as small as 500 microm were detectable with optical coherence tomography. The acquired images closely matched histologic images, demonstrating details at the level of mucosal layers, glands, alveoli, and respiratory bronchioles. CONCLUSIONS: Optical coherence tomography with near-infrared interferometric methods enables near real-time in vivo near-histologic resolution optical imaging. With further advances, optical coherence tomography has the potential for real-time accurate and early pleural and subpleural diagnostics by using small-diameter flexible fiberoptic endoscopic probes for a wide range of thoracic surgical applications. PMID- 15746747 TI - Does esophagogastric anastomotic technique influence the outcome of patients with esophageal cancer? AB - OBJECTIVE: We sought to compare the outcome of patients with esophageal cancer who had either modified Collard or standard hand-sewn cervical esophagogastric anastomoses in reconstruction during esophagectomy. METHODS: From March of 1996 to October of 2002, 274 patients with esophageal cancer underwent esophagectomy with gastric replacement and cervical esophagogastric anastomosis. Beginning in March of 2001, a modified Collard technique (stapled) was used in most patients (n = 86) for cervical esophagogastric anastomosis; a standard hand-sewn technique (sewn) was used in all others (n = 188). Using a propensity score based on 8 variables (age, gender, race, surgeon, surgical approach, pathologic stage, histologic cell type, and induction chemoradiotherapy), 85 patient pairs were matched and followed for time-related events. Outcome comparisons included cervical wound infection, cervical anastomotic leak, other hospital complications, length of stay, anastomotic dilatation, reflux symptoms, and survival. RESULTS: At 30 days, freedom from cervical wound infection was 92% for stapled versus 71% for sewn anastomoses ( P = .001), and freedom from cervical anastomotic leak was 96% versus 89% ( P = .09), respectively. Other hospital complications occurred in 58% and 49%, respectively ( P = .17). Median length of stay was 10 days for both ( P = .3). At 2 years, freedom from anastomotic dilatation was 34% for stapled versus 10% for sewn anastomoses ( P < .0001), and the mean number of dilatations per patient was 2.4 versus 4.1 ( P = .0001), respectively. Reflux was rare for both. Thirty-day, 6-month, and 24-month survivals were 98%, 91%, and 77% for stapled anastomoses and 98%, 88%, and 69% for sewn anastomoses ( P = .3). CONCLUSIONS: The modified Collard anastomotic technique dramatically reduces morbidity after esophagectomy. It should replace hand-sewn esophagogastric anastomoses. PMID- 15746748 TI - Feasibility and safety of airway bypass stent placement and influence of topical mitomycin C on stent patency. AB - OBJECTIVE: Airway bypass via transbronchial fenestration has been shown to improve forced expiratory volume and flow in explanted human emphysematous lungs. The aim of this study was to evaluate the feasibility and safety of in vivo airway bypass stent placement by using a canine model and to assess the influence of topical mitomycin C on the prolongation of stent patency. METHODS: With dogs under general anesthesia, suitable segmental and subsegmental bronchial wall sites were selected by direct visualization with a flexible bronchoscope. Peribronchial blood vessel injury was avoided by using a Doppler probe. Transbronchial fenestration was formed with a 22-gauge transbronchial needle, and the passage was then dilated with a 2.5-mm angioplasty balloon. A balloon expandable stainless-steel stent (3 mm long x 3 mm wide) with a sleeve of silicone rubber covering was placed within the fenestration. Animals were bronchoscoped weekly to assess stent patency. Seventy stents were placed in 12 dogs. Thirty-five served as controls, and the other 35 received transbronchoscopic topical application of mitomycin C once weekly to evaluate the effect on the maintenance of stent patency. Mitomycin C stents were divided into 4 groups according to the number of treatments: group A, 1 treatment only; group B, 4 weeks; group C, 7 weeks; and group D, 9 weeks. Each once-weekly mitomycin C application consisted of 0.2 mL at a concentration of 1 mg/mL, delivered through a small polyethylene catheter. RESULTS: Four instances of minor and brief bleeding occurred during stent placement and resolved without incident. One pneumothorax occurred and was treated by chest tube placement, without any adverse sequelae. There was no mortality associated with stent placement. No delayed hemorrhage or pneumothorax occurred. All control stents were occluded at the 1-week follow-up. The median durations of stent patency for group A (n = 8), group B (n = 9), group C (n = 10), and group D (n = 8) were 3, 8, 13, and greater than 20 weeks, respectively. CONCLUSIONS: Airway bypass stent placement can be performed safely. In an animal model, most stents became occluded within 1 week, but topical mitomycin application resulted in significant prolongation of patency. PMID- 15746749 TI - Radiofrequency ablation for the treatment of non-small cell lung cancer in marginal surgical candidates. AB - OBJECTIVE: Treatment options for patients with non-small cell lung cancer who are not surgical candidates or who refuse operation are limited. Radiofrequency ablation represents a potential less invasive option for these patients. Our initial experience with radiofrequency ablation for peripheral, primary non-small cell lung cancer is reported. METHODS: We treated 21 tumors in 18 patients. Median age was 75 (range 58-86) years. Cancer stages were I (n = 9), II (n = 2), III (n = 3), and IV (n = 4). Patients with stage IV disease included 3 with recurrence after previous lobectomies and 1 with a synchronous liver metastasis also treated with radiofrequency ablation. Median tumor diameter was 2.8 cm (range 1.2-4.5 cm). Radiofrequency ablation was delivered by minithoracotomy in 2 cases and by a computed tomography-guided percutaneous approach in 16 patients. Computed tomographic and positron emission tomographic scans were used to evaluate recurrence and radiographic response in ablated nodules. RESULTS: One postoperative death occurred from pneumonia after open radiofrequency ablation. Median hospital stay was 2.5 days. A chest tube or pigtail catheter was required in 7 patients (38.9%) for procedure-related pneumothoraces. At a median follow-up of 14 months, 15 patients (83.3%) were alive. Local progression occurred in 8 nodules (38.1%). Mean and median progression-free intervals were 16.8 and 18 months, respectively. For stage I cancers, mean progression-free interval was 17.6 months. Median progression-free interval was not reached. CONCLUSION: This study demonstrates the feasibility of radiofrequency ablation for small, peripheral non-small cell lung cancer tumors. Local control is comparable to, if not better than, that provided by radiotherapy. Radiofrequency ablation should continue to be evaluated by thoracic surgeons as a noninvasive therapy for the high-risk patient with non-small cell lung cancer. PMID- 15746750 TI - Emergency endovascular interventions for acute thoracic aortic rupture: four-year follow-up. AB - OBJECTIVE: High mortality and paraplegia rates associated with the surgical management of acute thoracic aortic ruptures limit its success. It was our objective to evaluate whether emergency endovascular interventions would improve the outcomes of these patients. METHODS: Sixty patients aged 28 to 83 years were admitted to our institution with an acute rupture of the thoracic aorta (27 ruptured aneurysms, 15 perforated type B dissections, 18 traumatic ruptures). Twenty-eight patients were treated surgically with cardiopulmonary bypass, and 32 patients were acutely treated with an endovascular stent graft. Medical records were reviewed for prehospital and emergency department data, operative findings, and outcomes. Patients were followed up at yearly intervals with high-resolution multidetector computed tomographic angiography. RESULTS: Perioperatively, there were 1 death (3.1%) among the 32 patients in the endovascular group and 5 deaths (17.8%) among the 28 patients in the surgical group. There were 4 late deaths in the endovascular group and 1 in the surgical group. There were 2 access failures in the endovascular group. There were 1 stroke in the endovascular group and 1 case of paraplegia in the surgical group. Three patients in the endovascular group had endovascular leaks develop that required reintervention. Two patients in the endovascular group had late thrombosis of the left subclavian artery. CONCLUSION: Despite encouraging early outcomes, midterm results suggest a trend toward increased reintervention and late complication rates in the endovascular group. Therefore continued surveillance of patients treated with stent grafts is necessary. PMID- 15746751 TI - Surveillance computed tomography after complete resection for non-small cell lung cancer: results and costs. AB - OBJECTIVE: We sought to determine the prevalence of defined abnormalities on surveillance computed tomography after complete resection for non-small cell lung cancer, as well as the nature and cost of further testing prompted by these abnormalities. We also sought to determine whether resectable metachronous lung cancer can be detected with surveillance scans. METHODS: A retrospective analysis was performed of all patients who presented for follow-up in 2002 after complete resection for non-small cell lung cancer. Data collected included demographics, clinicopathologic features of the initial lung cancer, the number and results of surveillance computed tomographic scans performed in 2002, the attending surgeons' impressions of the surveillance scans, the nature of any abnormalities and further diagnostic testing prompted by these abnormalities, and the nature of any lung cancer detected on surveillance scans, as well as the treatment rendered. The cost of surveillance scanning and associated diagnostics was computed by using Medicare fee schedules. RESULTS: Two hundred thirteen patients met the criteria for inclusion in the study cohort. One hundred sixty-eight surveillance scans were performed in 140 of these patients. One hundred five scans were interpreted as abnormal by the radiologist with regard to pulmonary nodules, adenopathy, or pleural fluid, but the surgeon was suspicious for recurrent or new primary lung cancer in only 32 of 105 scans. Further workup revealed recurrent or new primary lung cancer in 16 of 32 patients, with 6 undergoing resection for localized disease. The cost of the surveillance scans and associated care in the study cohort were 16.6% higher than the cost of care in a hypothetically identical cohort not subjected to surveillance scanning. CONCLUSIONS: Surveillance computed tomography is frequently abnormal after complete resection for non-small cell lung cancer; however, the majority of these abnormalities are not clinically suspicious. Resectable metachronous lung cancer is detected by using surveillance scanning; however, the use of this modality can be associated with increased cost. PMID- 15746752 TI - Repair of congenital heart lesions combined with lung transplantation for the treatment of severe pulmonary hypertension: a 13-year experience. AB - OBJECTIVE: In patients with severe pulmonary hypertension associated with congenital heart disease, we prefer to perform repair of the congenital heart disease and lung transplantation whenever feasible so as to augment the donor pool and avoid the cardiac complications associated with heart transplantation. We report our experience with repair of congenital heart disease and lung transplantation and compare the results with those of patients who underwent heart-lung transplantation during the same period. METHODS: The records of patients who had repair of congenital heart disease and lung transplantation (n = 35) and heart-lung transplantation (n = 16) between 1990 and 2003 were reviewed. RESULTS: The underlying congenital heart disease in the repair of congenital heart disease and lung transplantation group included transposition of great vessels (n = 2), atrioventricular canal defect (n = 2), ventricular septal defect (n = 9), pulmonary venous obstruction (n = 7), scimitar syndrome (n = 2), pulmonary arterial atresia or stenosis (n = 5), and others (n = 8). Thirteen of the patients undergoing repair of congenital heart disease and lung transplantation (37.1%) had the congenital heart disease repaired before lung transplantation; the remaining congenital heart disease repairs were performed concurrently with transplantation. Sixteen patients underwent heart-lung transplantation because of poor left ventricular function or single-ventricle anatomy. Freedoms from bronchiolitis obliterans at 1, 3, and 5 years were 72.9%, 54.7%, and 54.7% for the repair of congenital heart disease and lung transplantation group and 77.8%, 51.9%, and 38.9% for the heart-lung transplantation group, respectively. Survivals at 1, 3, and 5 years were 62.9%, 51.4%, and 51.4% for the repair of congenital heart disease and lung transplantation group and 66.5%, 66.5%, and 60% for the heart-lung transplantation group, respectively. CONCLUSION: Repair of congenital heart disease and lung transplantation is a feasible treatment option. Long-term outcome is determined by associated complications related to lung transplantation. Despite the complexity of combined congenital heart disease repair with lung transplantation and the resulting perioperative morbidity, the patients had similar outcomes to those of patients who underwent heart-lung transplantation. PMID- 15746753 TI - Historical perspectives of the American Association for Thoracic Surgery: Stuart Harrington, MD (1889-1975). PMID- 15746754 TI - Regurgitation through a stentless prosthesis treated with aortic root banding. PMID- 15746755 TI - Cardiac pheochromocytoma. PMID- 15746756 TI - Internal thoracic artery injury after transvenous pacemaker implantation. PMID- 15746757 TI - A method of using the pulmonary trunk to form a trileaflet valve. PMID- 15746758 TI - Stapled excision of the left atrial appendage. PMID- 15746759 TI - Acute mitral regurgitation caused by papillary muscle rupture in the immediate postpartum period revealing Ehlers-Danlos syndrome type IV. PMID- 15746760 TI - Endovascular treatment of right-sided subclavian artery aneurysm in a congenitally malformed aortic arch. PMID- 15746761 TI - A successful total cavopulmonary connection conversion 13 years after an anatomic repair for tricuspid atresia IIc with severe pulmonary resistance. PMID- 15746762 TI - Could multidetector computed tomography play a role in the management of patients readmitted with recurrent chest pain after surgical coronary revascularization? PMID- 15746763 TI - Unusually located intrathoracic extrapulmonary mediastinal hydatid cyst manifesting as Pancoast syndrome. PMID- 15746764 TI - Video-assisted mediastinoscopy: a useful technique for paratracheal mesothelial cysts. PMID- 15746765 TI - Successful closure of an empyema space associated with persistent bronchopleural fistula: Muscle transposition combined with endobronchial plug. PMID- 15746766 TI - Extended right pneumonectomy with partial left atrial resection for primary leiomyosarcoma of the mediastinum. PMID- 15746767 TI - Vascular endothelial growth factor before cells. PMID- 15746773 TI - Conquering absolute contraindications to transplantation: positive-crossmatch and ABO-incompatible kidney transplantation. PMID- 15746775 TI - Expert witness testimony: the problem and recommendations for oversight and reform. PMID- 15746776 TI - Flaws in methods of evidence-based medicine may adversely affect public health directives. PMID- 15746778 TI - Endovascular abdominal aortic aneurysm repair is more profitable than open repair based on contribution margin per day. AB - BACKGROUND: Earlier studies have reported that endovascular abdominal aortic aneurysm (EAAA) repair yields lower total profit margins than open AAA (OAAA) repair. This study compared EAAA versus OAAA based on contribution margin per day, which may better measure profitability of new clinical technologies. Contribution margin equals revenue less variable direct costs (VDCs). VDCs capture incremental resources tied directly to individual patients' activity (eg, invoice price of endograft device, nursing labor). Overhead costs factor into total margin, but not contribution margin. METHODS: The University of Michigan Health System's cost accounting system was used to extract fiscal year 2002-2003 information on revenue, total margin, contribution margin, and duration of stay for Medicare patients with principal diagnosis of AAA (ICD-9 code 441.4). RESULTS: OAAA had revenues of $37,137 per case versus $28,960 for EAAA, similar VDCs per case, and thus higher contribution margin per case ($24,404 for OAAA vs $13,911 for EAAA, P < .001). However, OAAA had significantly longer mean duration of stay per case (10.2 days vs 2.2 days, P < .001). Therefore, mean contribution margin per day was $2948 for OAAA, but $8569 for EAAA ( P < .001). CONCLUSIONS: On the basis of contribution margin per day, EAAA repair dominates OAAA repair. The shorter duration of stay with EAAA allows higher throughput, fuller overhead amortization, better use of scarce inpatient beds, and higher health system profits. Surgeons must understand overhead allocation to devices, especially when new technologies cut duration of stay markedly. PMID- 15746779 TI - The European Working Time Directive: One for all and all for one? AB - BACKGROUND: The European Working Time Directive (EWTD) became law in Britain on October 1, 1998. As a result, the maximum period that may be spent as a resident in hospitals is 56 hours per week and after August 2009, 48 hours per week. The aim of this study was to determine the views of senior house officers (SHOs), specialist registrars (SpRs), and general consultant surgeons (CONs) in Wales on the influence of the EWTD on surgical training and clinical experience. METHODS: In this cohort study, a postal questionnaire was sent to 150 SHOs in surgical specialties, 50 general surgical SpRs, and all 84 CONs in the Welsh Deanery. RESULTS: The response rates were 81%, 78%, and 71% for SHOs, SpRs, and CONs, respectively. The vast majorities at all grades (88% SHOs, 100% SpRs, and 96% CONs) were unhappy with the introduction of EWTD legislation to clinical medicine. Most felt that EWTD legislation will have a negative effect on clinical experience (96% SHOs, 97% SpRs, 96% CONs); patient care (83% SHOs, 85% SpRs, 96% CONs); and training (94% SHOs, 100% SpRs, 93% CONs). Furthermore, a large proportion felt surgical training should be exempt from EWTD regulations (76% SHOs, 87% SpRs, 89% CONs). A significant proportion at each grade was opposed to the introduction of shifts in order to comply with regulations (78% SHOs, 87% SpRs, 89% CONs), and an alarming number have considered leaving the National Health Service when the regulations are enforced (29% SHOs, 41% SpRs, 33% CONs). CONCLUSION: This study shows that, in Wales at least, a vast majority of surgical trainees and consultants alike are opposed to the introduction of the EWTD and believe it will have a detrimental effect on training, patient care, and doctors' lives outside of medicine. PMID- 15746780 TI - Tumor versus neoplasm: Isn't it time to use the correct term--neoplasm? PMID- 15746781 TI - Electronic tagging of surgical sponges to prevent their accidental retention. AB - BACKGROUND: The inadvertent "loss" of surgical sponges, towels, and instruments remains an unsolved problem. The means of preventing this mishap involve an inventory of all implements going in and coming out of the surgical wound, supplemented by a careful search. This work describes a different approach to this problem: tagging articles placed into the wound with markers able to be rapidly detected electronically. METHODS: Surgical sponges were tagged with either active targets or inert, dummy targets and placed in different locations within the body of 50 cadavers. Using a device we constructed, blinded readings were performed to test the accuracy of detecting retained sponges. RESULTS: A total of 600 readings were made in 50 cadavers, 300 of active targets and 300 of inert, dummy targets, indistinguishable in appearance. The active markers were detected with 100% accuracy. There were no false positive detections of the dummy tags. CONCLUSION: The present work demonstrates the feasibility of detecting retained surgical sponges by electronic means blinded to whether the tag contained an active or inert marker. PMID- 15746782 TI - Invited commentary: Making patient safety automatic. PMID- 15746783 TI - Editor's note: Retained foreign bodies--why do we still allow them to occur? PMID- 15746784 TI - Sharp liver transection versus clamp crushing technique in liver resections: a prospective study. AB - BACKGROUND: Parenchymal liver transection constitutes an important phase of liver resection. Serious intraoperative bleeding, together with injuries to vital structures of the liver remnant, can occur during this stage. A method of sharp liver parenchymal transection with scalpel is compared in a prospective randomized manner with the widely used clamp crushing technique. METHODS: Patients scheduled for hepatectomy under selective hepatic vascular exclusion (N = 82) were allocated randomly to either the sharp transection group (n = 41) or the clamp crushing group (n = 41). Warm ischemic time, blood loss and transfusions, postoperative morbidity and mortality, and tumor-free margins were recorded in both groups and analyzed. RESULTS: When the sharp transection group was compared with the clamp crushing group, the two groups were similar in warm ischemic time (median 36 vs 34 minutes), total operative time (median 205 vs 211 minutes), intraoperative blood loss (median 500 vs 460 mL), blood transfusion requirements (median value 0 in both groups), and overall complication rate (44% vs 39%). However, sharp transection yielded better tumor-free margins compared with the clamp crushing technique (12 +/- 1.4 mm vs 8 +/- 1.5 mm, mean +/- SD, P < .05). CONCLUSION: Sharp liver parenchymal transection with a scalpel is equally safe in terms of blood loss and mortality compared with the clamp crushing method. Although it is a technically demanding method, requiring selective hepatic vascular occlusion, it may be recommended when the tumor-free margins are anticipated to be narrow. PMID- 15746785 TI - Surgical management and long-term outcome of complicated liver hydatid cysts caused by Echinococcus granulosus. AB - BACKGROUND: The aim of this retrospective study was to evaluate clinical presentation and long-term outcome of patients treated surgically for complicated liver hydatid cysts. PATIENTS AND METHODS: Eighty-four patients with liver hydatid cysts underwent an operation at the Geneva University Hospital between 1980 and 1999. Clinical presentation, postoperative morbidity, mortality, and long-term recurrence rate were evaluated. RESULTS: Among the 84 patients with liver hydatid disease, 35 patients (41%) presented complicated cysts (ie, cysts that had developed a fistula into adjacent structures or organs). In most patients, the fistula communicated with the biliary tree (n = 25), but we also observed communication with the right lung (n = 3), the right diaphragm (n = 2), liver parenchyma (n = 1), and peritoneal cavity (n = 1). Complete removal of the cystic disease was possible in 24 of 35 patients (70%). In 11 patients, fragments of cysts were not removed because of their location adjacent to main vessels. Postoperatively, 8 patients (23%) developed a severe complication (grade II and III). There were no postoperative deaths, and no recurrences of hydatid disease were observed with a median follow-up of 8.6 years (complete follow-up was obtained in 69% of patients). CONCLUSIONS: Complicated liver hydatid disease is frequent and was observed in almost half of patients operated for liver hydatid cysts at our center. Using a surgical strategy aimed at complete removal of cystic and pericystic tissue with simultaneous treatment of the fistulous tract, we observed 23% postoperative morbidity, no mortality, and no recurrence of disease with a median follow-up of >8 years. PMID- 15746786 TI - Laparoscopic assisted distal gastrectomy for early gastric cancer: Five years' experience. AB - BACKGROUND: Laparoscopic assisted gastrectomy is being reported increasingly as the treatment of choice for early gastric cancer. However, no reports concerning the prognosis of patients who have undergone laparoscopic assisted distal gastrectomy (LADG) for early gastric cancer or data comparing the results to those obtained after open gastric surgery are yet available. METHODS: A retrospective study was performed comparing laparoscopic assisted and open distal gastrectomies for early gastric cancer. Eighty-nine patients who underwent LADG were compared to 60 who underwent conventional open distal gastrectomy (DG) in terms of pathologic findings, operative outcome, complications, and survival. RESULTS: There were no significant differences between LADG and DG in operation time (209 vs 200 minutes), complication rate (9% vs 18%), and 5-year survival rate (98% vs 95%). There were differences between LADG and DG with regard to blood loss (237 vs 412 mL), number of lymph nodes (19 vs 25), postoperative stay (17 vs 25 days), and the duration of epidural analgesia (2 vs 4 days) ( P < .05 each). CONCLUSIONS: For properly selected patients, LADG can be a curative and minimally invasive treatment for early gastric cancer. PMID- 15746787 TI - Management and outcome of hemorrhage due to arterial pseudoaneurysms in pancreatitis. AB - BACKGROUND: Arterial pseudoaneurysm formation in pancreatitis is a rare complication. The optimal treatment modality is controversial. Operative treatment and interventional treatment, either alone or as a temporizing method with a later operation, are options. METHODS: In this single-center, patient based cohort study, we managed 35 patients (8 with necrotizing pancreatitis and 27 with chronic pancreatitis) with bleeding pseudoaneurysms treated over a period of 10.5 years with a median follow-up of 4.6 years. Angiography was performed depending on the patient's hemodynamic condition. RESULTS: Angiography had a sensitivity of 96% for 26 patients. Angiographic embolization as primary treatment was performed in 16 patients (61% embolization rate); there were 2 rebleeding complications. No patients required intervention for embolization complications after discharge. Nineteen patients (54%) underwent an operation, 9 urgently without angiographic evaluation. The overall mortality rate for the 35 patients was 20% (19% for embolization, 21% after an operation). For necrotizing pancreatitis, an advantage of angiographic embolization was observed (mortality in 2/5 vs 2/3 after surgery). Ligation or repair of the bleeding vessel was complicated by higher rebleeding rates (6/13) than partial pancreatectomy (1/6). CONCLUSIONS: Concerns that angiographic embolization is unable to provide definitive hemostasis in both acute and chronic pancreatitis are unfounded. In the operative treatment of chronic pancreatitis, partial pancreatectomy is superior to vessel ligation, depending on the patient's general condition and degree of pancreatic inflammation. We propose an algorithm for the management of arterial pseudoaneurysms in the setting of pancreatitis. PMID- 15746788 TI - Rectovaginal fistulas after rectal cancer surgery: Incidence and operative repair by gluteal-fold flap repair. AB - BACKGROUND: We investigated the correlation between operative procedures for rectal carcinoma and postoperative rectovaginal fistulas (RVF), and treatment for RVF. METHODS: The medical records of 161 female patients with rectal carcinoma were examined retrospectively with respect to the cause, incidence, and methods of treatment for RVF occurring after rectal cancer operations, and to the outcomes of gluteal-fold flap repairs for RVF. RESULTS: Of the 161 patients, 16 developed RVF clinically. The incidence of RVF was significantly higher in patients who were anastomosed by the double stapling technique (DST) and had concomitant resection of the vaginal wall. No statistical difference was found between the established diverting ostomy group and the no-stoma group. Six patients recovered by the establishment of a diverting ostomy only. The gluteal fold flap technique was performed for 5 patients. No RVF recurrences were noted in these 5 patients. CONCLUSIONS: The incidence of RVF was higher in the patients who were anastomosed by DST or had concomitant resection of the vaginal wall. Although some RVFs heal with only fecal diversion, for patients in whom RVF is caused by involvement of the vaginal wall in the circular staple or intersphincteric resection, good results are obtained with the gluteal-fold flap repair technique. PMID- 15746789 TI - A randomized, prospective, parallel group study comparing the Harmonic Scalpel to electrocautery in thyroidectomy. AB - BACKGROUND: Exhaustive hemostasis is essential to successful thyroidectomy. Electrocoagulation to control bleeding has the potential risk of injuring the surrounding structures from lateral dispersion of heat. The Harmonic Scalpel (HS) cuts and coagulates simultaneously using mechanical vibration rather than high temperatures. Because its use in thyroidectomies has been limited, we sought to compare procedure parameters and complications of thyroidectomies performed using the HS with those using electrocoagulation. METHODS: Sixty patients were randomized into 2 surgical groups, HS and the standard technique using electrocautery and ligatures as the primary hemostatic method. A sample t test or Wilcoxon rank sum test was used to compare the following parameters: operative time, number of ligatures, blood loss, pain intensity, need for extra dose(s) of analgesic, incidence of recurrent laryngeal nerve palsy, and hypoparathyroidism. RESULTS: Both groups of 30 patients were comparable in age, gender, and nature of disease. In 38 patients (63%), surgery was performed for benign disease, and in 22 (37%), for differentiated carcinoma. Operative time (arithmetic mean +/- SD) was 25 minutes less in the HS group (96 +/- 23 vs 121 +/- 34, P = .005). Median number of ligatures in the HS group was 1 (range, 0-7) versus 17 (range, 6-28) ( P < .001). Mean blood loss, estimated by gauze weight, was less with HS (35 +/- 27 mL vs 54 +/- 51 mL, P = .06). Drainage during the first 24 postoperative hours and pain intensity during the first postoperative week were similar in both groups. There were no episodes of persistent nerve palsy or hypoparathyroidism in either group. CONCLUSION: The use of HS in thyroidectomies requires less operative time than does electrocoagulation. PMID- 15746790 TI - Recurrent laryngeal nerve palsy after thyroidectomy with routine identification of the recurrent laryngeal nerve. AB - BACKGROUND: The aim of this study was to assess the risk of recurrent laryngeal nerve palsy (RLNP) after thyroidectomy with routine identification of the recurrent laryngeal nerve (RLN) during the operation. METHODS: The present study was confined to 521 patients, 348 total lobectomies and 178 total thyroidectomies, treated by the same surgeon. Temporary and permanent RLNP rates were analyzed for patient groups with stratification of primary operation for benign thyroid disease, thyroid cancer, Graves' disease, and reoperation. Measurement of the RLNP rate was based on the number of nerves at risk. Twenty six RLNs in 20 thyroid cancer patients with intentional sacrifice were excluded from analysis. RESULTS: Forty RLNs (40 patients) developed postoperative RLNP. Complete recovery of RLN function was documented for 35 of the 37 patients (94.6%) whose RLN integrity had been ensured intraoperatively. Recovery from temporary RLNP ranged from 3 days to 4 months (mean, 30.7 days). Overall incidence of temporary and permanent RLNP was 5.1% and 0.9%, respectively. The rates of temporary/permanent RLNP were 4.0/0.2%, 2.0/0.7%, 12.0/1.1%, and 10.8/8.1% for groups classified according to benign thyroid disease, thyroid cancer, Graves' disease, and reoperation, respectively. CONCLUSIONS: Operations for thyroid cancer, Graves' disease, and recurrent goiter demonstrated significantly higher RLNP rates. Invasion of RLN was identified in 19.4% of patients with thyroid cancer. Postoperatively, the RLN recovered in most of the patients without documented nerve damage during the operation. Total lobectomy with routine RLN identification is recommended as a basic procedure in thyroid operations. PMID- 15746791 TI - Icodextrin reduces postoperative adhesion formation in rats without affecting peritoneal metastasis. AB - BACKGROUND: Peroperative peritoneal trauma activates a cascade of peritoneal defense mechanisms responsible for postoperative adhesion formation. The same cascade seems to play a role in the process of intra-abdominal tumor recurrence. Icodextrin is a glucose polymer solution that is absorbed slowly from the peritoneal cavity, allowing prolonged "hydroflotation" of the viscera, thereby decreasing adhesion formation. This study evaluated the adhesion-preventing properties of icodextrin and its effect on peritoneal metastasis. METHODS: Reproducible rat models of peritoneal trauma were used, allowing semiquantitative scoring of adhesion formation or tumor load. In one experiment, peritoneal trauma was inflicted; one group was treated by peroperative intra-abdominal instillation of 7.5% icodextrin, one by instillation of RPMI (placebo), and one had no instillate (controls). In another experiment involving a different model of peritoneal trauma, the coloncarcinoma cell line CC531 was injected intraperitoneally to induce tumor load, again using these three groups. RESULTS: Treatment of peritoneally traumatized rats with icodextrin caused a 51% reduction in postoperative adhesion formation ( P < .001). However, peroperative intra abdominal treatment with icodextrin did not affect intraperitoneal tumor cell adhesion and growth of free intra-abdominal tumor cells in rats with this model of severe peritoneal trauma. CONCLUSION: A 7.5% icodextrin solution is effective in reducing postoperative adhesions without promoting tumor recurrence and therefore may prove useful and safe in oncologic surgery. PMID- 15746792 TI - Metalloproteinases and their inhibitors are markers of plaque instability. AB - BACKGROUND: Our aim was to investigate the release, activity, and expression of matrix metalloproteinases (MMPs)-1, -2, -3 and -9, and tissue inhibitors of metalloproteinases (TIMPs)-1 and -2 in patients undergoing carotid endarterectomy and to determine whether altered plasma levels of MMPs and TIMPs may be correlated with carotid instability. METHODS: The carotid plaques of 53 consecutive patients who underwent carotid endarterectomy were classified histologically as stable or unstable. The release of MMPs and TIMPs was analyzed in the serum of patients with stable and unstable carotid plaques, and in 15 age matched healthy volunteers. The production, activity, and expression of MMPs and TIMPs were determined by Western blotting, zymography, and reverse transcriptase polymerase chain reaction in the carotid specimens. RESULTS: Twenty-nine (55%) patients had an unstable carotid plaque and 24 (45%) a stable plaque. Plasma levels of MMPs were higher in patients with unstable plaques compared to patients with stable plaques and healthy volunteers ( P < .001), whereas plasma levels of TIMPs were lower in patients with unstable plaques compared to patients with stable plaques and healthy volunteers ( P < .001). In the carotid specimens, we found increased activity, production, and expression of MMPs, and decreased activity, production and expression of TIMPs in unstable plaques compared to stable plaques ( P < .001). After endarterectomy, plasma levels of MMPs and TIMPs in patients with unstable and stable plaques returned to the values found in healthy volunteers. CONCLUSIONS: Our study demonstrated that an imbalance exists between MMPs and TIMPs in unstable carotid plaques, which is reflected in the plasma levels of these markers. These data may help in selecting patients at high risk for cerebral events. PMID- 15746794 TI - Internal herniation concurrent with ileosigmoid knotting or sigmoid volvulus: Presentation of 12 patients. AB - BACKGROUND: Internal herniation concurrent with ileosigmoid knotting or sigmoid volvulus is an unusual and complex form of closed-loop obstruction that may result in a fatal outcome unless treated timely and properly. The aim of this article was to review our experience with this condition, with emphasis on the etiopathogenesis, clinicopathologic features, and treatment options. METHODS: We conducted a retrospective analysis of medical records of 12 patients treated at 2 university hospitals over a period of 30 years between 1970 and 2000. RESULTS: In this series, the internal herniation resulted in ileosigmoid knotting in 8 cases, whereas it was concomitant with sigmoid volvulus in 4 cases. The types of internal herniation were identified as transmesenteric through the Treves field in 8 patients and as transomental, intersigmoidal, pericecal, and around omphalomesenteric fibrous cord in 1 patient each. The rate of gangrenous bowel was 100%. En bloc resection for combined gangrene of small bowel and large bowel was the treatment of choice in 7 patients, of whom 5 underwent the Hartmann's procedure and 2 underwent primary sigmoidectomy-anastomosis in addition to primary enterectomy-anastomosis. Primary sigmoidectomy-anastomosis and Mikulicz's procedure were performed in 2 patients for gangrenous sigmoid colon only. Three patients underwent primary enterectomy-anastomosis for gangrenous small bowel only. The morbidity rates and the mortality rate were both 33.3%. The mean length of hospital stay following emergency operations was 11.2 days. CONCLUSIONS: In particular, surgeons who are from developing countries that form the world's "volvulus belt" should be aware of this entity's features and be ready to perform an appropriately selected surgical option for a given patient to accomplish the optimal clinical outcome. PMID- 15746793 TI - Randomized prospective trial of early steroid withdrawal compared with low-dose steroids in renal transplant recipients using serial protocol biopsies to assess efficacy and safety. AB - BACKGROUND: Corticosteroid therapy after renal transplantation is associated with many adverse effects. Newer immunosuppressive agents may allow for safe and effective reductions in dose or early steroid withdrawal. METHODS: In this prospective, single-center clinical trial, 60 patients were randomized into 2 groups: control patients (n = 28), who received low doses of prednisone throughout, and study patients (n = 32), who were withdrawn from steroids 7 days posttransplant. Patients received a limited course of rabbit antilymphocyte globulin (rALG) induction therapy, tacrolimus (TAC), and mycophenolate mofetil (MMF). Patients were followed for clinical outcomes and renal function. Protocol biopsies were performed at 1, 6, and 12 months. RESULTS: Clinical rejections occurred in 11% of controls and 13% of study patients. Renal function was well maintained and equivalent in both groups. In all, 111 protocol biopsies were performed without complications. Subclinical rejection was noted in only 2 protocol biopsies, and borderline changes were seen in 12 biopsies, all of which were distributed equally between both groups. Unsuspected acute TAC toxicity was seen in 8 biopsies. Protocol biopsies led to changes in therapy in 10% of patients. In both groups, serial protocol biopsies demonstrated increased allograft fibrosis over time, which was significant at 1 year in the steroid withdrawal group. CONCLUSION: The immunosuppressive combination of rALG, TAC, and MMF prevents subclinical rejection and the need for high doses of steroids after transplantation. However, continual low-dose steroid therapy may aid in preventing chronic allograft fibrosis. Protocol biopsies help define the short term and long-term risks of steroid withdrawal therapy. PMID- 15746795 TI - Schwannoma of the median nerve: Iatrogenic injury after axillary lymph node biopsy. PMID- 15746796 TI - Truth-telling about terminal diseases. PMID- 15746797 TI - Superior mesenteric artery syndrome after laparoscopic Roux-en-Y gastric bypass. PMID- 15746799 TI - Pancreatic renal cell carcinoma metastasis presenting with upper gastrointestinal bleeding. PMID- 15746800 TI - Monolateral mature ovarian cystic teratoma. PMID- 15746801 TI - Have you noticed? PMID- 15746802 TI - Re: Residency training in surgery in the 21st century: A new paradigm. PMID- 15746813 TI - Evidence-based practice: a core competency for dietetics. PMID- 15746814 TI - Nutrition quality and education in K-12 schools. PMID- 15746815 TI - The safety of beef in the United States. PMID- 15746816 TI - Preparing for reentry: handling career interruptions. PMID- 15746817 TI - Dietetics professionals' role in the changing face of America's health care: the chronic care model. PMID- 15746819 TI - Contaminated oats and other gluten-free foods in the United States. PMID- 15746820 TI - Ancel Keys, seven countries study, China connections, and K rations. PMID- 15746821 TI - A few more thoughts about fish and fish oil. PMID- 15746822 TI - Dietary intake estimates in the National Health Interview Survey, 2000: methodology, results, and interpretation. AB - OBJECTIVES: To describe the implementation of the Multifactor Screener in the 2000 National Health Interview Survey (NHIS); to provide intake estimates for fruits and vegetables, fiber, and percentage of energy from fat by various demographic and behavioral characteristics; and to discuss the strengths and weaknesses of the method. DESIGN/SUBJECTS: The 2000 NHIS was conducted in 38,632 households in a clustered sample representative of the 48 contiguous states in the United States. The Cancer Control Module was administered to adults (18 years and older) and included 17 dietary intake questions. ANALYSES: After excluding individuals with missing data or unlikely values on the diet questions, we computed individual intake of servings of fruits and vegetables, percentage of energy from fat, and grams of fiber. We estimated median intakes and distributions of intakes using adjusted variance estimates. We present bivariate relationships between diet and demographics and diet and behavioral characteristics in almost 30,000 adults in the United States. RESULTS: In general, intakes of these dietary factors were closer to recommendations among well-educated individuals, those engaged in other healthful behaviors, and underweight and normal weight individuals. Latinos had higher intakes of fruits and vegetables (men: 6 servings; women: 4.8 servings) and fiber (men: 23 g; women: 17 g), and generally a lower percentage of energy intake from fat (men: 33.7%; women: 32.1%) than did non-Latino whites (men: 5.4 servings; women: 4.5 servings; men: 19 g; women: 14 g; men: 33.9%; women: 32.0%) and non-Latino blacks (men: 5.4 servings; women: 4.4 servings; men: 19 g; women: 13 g; men: 34.7%; women: 33.5%). The strengths and limitations of the short dietary assessment method are discussed. CONCLUSIONS: The Multifactor Screener in NHIS, when used in conjunction with external reference data, provides reasonable estimates for three dietary factors and suggests relationships between intakes and other characteristics that are consistent with other data. Thus, these NHIS estimates could provide useful national benchmarks for local surveys using the same instrument. PMID- 15746823 TI - Anthropometric and psychosocial changes in obese adolescents enrolled in a Weight Management Program. AB - OBJECTIVE: To determine short- and long-term effects of the Bright Bodies Weight Management Program on obese adolescents and to further observe if a diet or nondiet approach is more successful. DESIGN: Twenty-five obese adolescents completed a 1-year, comprehensive weight-management program and returned for a 2 year follow-up. Adolescents were 11 to 16 years old (17 female, eight male) with mixed ethnic backgrounds. Although the program emphasizes a nondiet approach, eight children requested a structured meal plan (diet approach), while 17 were taught to make better food choices (nondiet approach). Body mass index (BMI) z score, body fat percent, and self-concept were measured at 0, 1, and 2 years. Outcomes were analyzed for the entire group and by diet method groups. STATISTICAL ANALYSIS: Changes in outcome variables were evaluated using covariance pattern models for repeated measures. RESULTS: At 1 year, the entire group (N=25) demonstrated a decrease in BMI z score (P <.001) and body fat percent (P <.001), while self-concept scores increased (P <.001). At 2 years, the decrease in BMI z score was still significant (P =.004) and body fat percent and self-concept scores remained improved, although not significant compared to baseline (P =.15 and P =.10, respectively). When comparing dietary approaches, the dieting group (n=8) tended to show favorable results short-term for BMI z score at year 1 (P =.11), but by year 2, the nondieting group (n=17) further improved BMI z score (P =.006), while the dieting group reverted toward baseline. CONCLUSIONS: The Bright Bodies Weight Management Program was successful at decreasing BMI z scores both short and long term. In a separate analysis, dieting showed more superior short-term results, but a nondiet approach demonstrated improved long-term results. PMID- 15746824 TI - Effects of an intensive diet and physical activity modification program on the health risks of adults. AB - BACKGROUND: This study assessed the clinical impact of lifestyle change education on chronic disease risk factors within a community. DESIGN: Randomized clinical trial. SETTING/PARTICIPANTS: Participants included 337 volunteers age 43 to 81 years from the Rockford, IL, metropolitan area. INTERVENTION: The intervention group attended a 40-hour educational course delivered over a 4-week period. Participants learned the importance of making healthful lifestyle choices and how to make improvements in nutrition and physical activity. MAIN OUTCOME MEASURES: Changes in health knowledge, nutrition, and physical activity behavior, and several chronic disease risk factors were assessed at baseline and 6 weeks. RESULTS: Beneficial mean changes in scores tended to be significant for the intervention group but not for the control group. Variables with improved scores included health knowledge, percent body fat, total steps per week, and most nutrition variables. Clinical improvements were seen in resting heart rate, total cholesterol, low-density lipoprotein cholesterol, and systolic and diastolic blood pressure. The control group experienced comparatively small but significant improvements in health knowledge, systolic and diastolic blood pressure, glucose, and in some nutrition variables. For almost all variables, the intervention group showed significantly greater improvements. CONCLUSIONS: This lifestyle modification program is an efficacious nutrition and physical activity intervention in the short term and has the potential to dramatically reduce the risks associated with common chronic diseases in the long term. PMID- 15746825 TI - Achieving substantial changes in eating behavior among women previously treated for breast cancer--an overview of the intervention. AB - OBJECTIVE: To describe the intervention in a clinical trial examining the effect of a plant-based diet on breast cancer recurrence. To report baseline to 12-month dietary change and investigate whether cooking-class attendance influenced adherence to the study's dietary targets. DESIGN: A descriptive analysis of baseline and 12-month dietary intake data and other variables from a subcohort of participants in the Women's Healthy Eating and Living Study. SUBJECTS/SETTING: Seven hundred thirty-nine women (primarily non-Hispanic white and well educated) who had been treated for early stage breast cancer. All were intervention group participants and had adhered to the Women's Healthy Eating and Living Study counseling and dietary assessment protocols. Mean age at study entry was 54 years, and mean body mass index was 26.7. INTERVENTION: Telephone counseling, complemented by an orientation meeting, cooking classes, and newsletters. MAIN OUTCOME MEASURES: The change in intake of vegetables, vegetable juice, fruit, fiber, and fat between baseline and 12 months is reported, and the association between cooking classes attended and overall dietary adherence is examined. STATISTICAL ANALYSES PERFORMED: Mean intake for vegetables, vegetable juice, fruit, fiber, and fat were calculated. Percentage of women meeting select Healthy People 2010 objectives were tabulated. RESULTS: Total daily vegetable, vegetable juice, fruit, and fiber intake increased significantly (P <.01), while fat decreased significantly (P <.01). The percentage of women meeting the Healthy People 2010 fruit and vegetable objectives increased substantially. Overall dietary adherence was associated with increased cooking-class attendance (P for trend <.01). CONCLUSIONS: A multimodal approach to dietary modification, based largely on individualized telephone counseling, can substantially change the overall dietary pattern of women previously treated for breast cancer. PMID- 15746826 TI - Food nutrition label use is associated with demographic, behavioral, and psychosocial factors and dietary intake among African Americans in North Carolina. AB - OBJECTIVE: Reading nutrition labels on food packages may improve food choices and enable healthful dietary practices. This report describes the prevalence of nutrition label use and its association with demographic, behavioral, and psychological factors and diet among African-American adults. DESIGN/SUBJECTS: Self-reported data from a population-based cross-sectional survey of 658 African Americans, aged 20 to 70 years, in North Carolina. An 11-page questionnaire assessed nutrition label use, fruit and vegetable consumption, total and saturated fat intakes, fat-related dietary behaviors, diet-related psychosocial factors, and demographic and behavioral characteristics. STATISTICAL ANALYSIS: Chi2 tests and logistic regression analyses examined associations of demographic, psychosocial, and behavioral factors with nutrition label use. Linear regression was used to estimate the variation in diet explained by label use. RESULTS: The mean age of participants was 43.9+/-11.6 years, 41% were men, 37% were college graduates, and 75% were overweight/obese. Seventy-eight percent of respondents read nutrition labels when they purchased packaged foods. Nutrition label use was significantly higher among participants who were women, older, educated beyond high school, and obese (P <.05). After adjusting for demographic characteristics, the strongest psychosocial predictors of nutrition label use were healthful eating self-efficacy, strong belief in a diet-cancer relationship, and trying to lose weight. Usual/often label users had higher fruit and vegetable consumption and lower fat intakes (P <.001), and nutrition labels explained 2% to 17% of the variance in dietary intake. CONCLUSIONS: Nutrition information on packaged foods appears to be a useful way to conduct point-of-purchase nutrition education among African Americans in North Carolina. Most respondents used food labels at least sometimes, but only about half usually or often did so. Efforts should be made to determine how all consumers could use nutrition labels effectively. PMID- 15746828 TI - Heritage retention and bean intake correlates to dietary fiber intakes in Hispanic mothers--Que Sabrosa Vida. AB - OBJECTIVE: To determine the correlation between fruit/vegetable and bean stage of change and heritage retention with fiber intake in Hispanic mothers. DESIGN: Cross-sectional study design used baseline survey data from the Que Sabrosa Vida nutrition intervention developed by the University of Texas School of Public Health, Human Nutrition Center (Houston, TX). Participants were recruited and data gathered from telephone surveys using random digit dial methodology ensuring proportional representation of demographic subgroups. SUBJECTS: Hispanic mothers (N=293) responded to the baseline survey in El Paso, TX. ANALYSES: Using SPSS version 11.0 (SPSS Inc, Chicago, IL), descriptive statistics, chi2 , correlations, and analyses of variance were used to determine the relationship of variables to fiber intake. Potential confounding variables included were grain stage of change, age, education level, and marital status. RESULTS: Mean fiber intake for mothers was 21.9 g/day and mean age was 35.8 years. Chi2 of fruit/vegetable stage was P =.784. Chi 2 of bean stage, heritage retention score, age, and education to fiber intake were significant, P =.008, P =.004, P =.005, and P =.001, respectively. Bean stage, heritage retention score, age, and education level were significantly correlated. One-way analysis of variance of bean stage, heritage retention score, age, and education level by fiber intake were significant, P =.006, P =.001, P =.004, and P =.002, respectively. CONCLUSIONS: Older Hispanic mothers in the action/maintenance bean stage with strong heritage retention score had greater fiber intakes than younger mothers in pre-action bean stage with weak heritage retention score. As education level increased fiber intake decreased. Encouragement of bean consumption and heritage retention may increase fiber intake in this Hispanic mother population. PMID- 15746830 TI - Impact of core and secondary foods on nutritional composition of diets in Native American women. AB - OBJECTIVE: To identify the core and secondary foods among Native-American women in Oklahoma and to determine their impact on nutrient and Food Guide Pyramid serving intakes. DESIGN: This descriptive study explored food intakes from 4-day weighed food records. Nutrient intakes were estimated using reference data used in national survey data. SUBJECTS/SETTING: Seventy-one Native-American women receiving services from three tribal health clinics in northeast Oklahoma. Statistical analyses performed A food-use frequency score was computed using frequencies of individuals consuming foods across each of 4 days of records. Leading contributors of nutrients and Food Guide Pyramid servings were identified from core and secondary foods. RESULTS: Thirty foods comprised the list of core foods, led by soda, coffee, and white bread. A majority of total energy, fat, saturated fat, monounsaturated fatty acids, polyunsaturated fatty acids, cholesterol, carbohydrate, calcium, vitamin C, folate, discretionary fat, and added sugar were derived cumulatively from the core and secondary foods. Forty percent of fruit Food Guide Pyramid servings were accounted for by two core foods, bananas, and orange juice. More than half of meat and vegetable Food Guide Pyramid servings were derived from core and secondary foods. CONCLUSIONS: Food patterning data are helpful in the development of effective nutrition education programs. We identified less nutrient-dense core foods that are contributing to discretionary fat and added sugar intakes. Targeted nutrition education programs for Native Americans should promote the nutrient-dense core and secondary foods, such as whole-wheat bread and fruit, while providing more healthful food alternatives to less nutrient-dense foods. PMID- 15746831 TI - A look at the educational preparation of the health-diagnosing and treating professions: do dietitians measure up? AB - Basic educational requirements for dietitians were developed almost 80 years ago and remain largely unchanged. In the interim, other health professions have increased their academic standards. A review of the educational preparation of 16 health-diagnosing and treating professions was undertaken to better understand the standards for dietetics education within a larger context. Educational standards for each profession were obtained and reviewed for types of degrees; duration of post-secondary, college-level education; division of didactic and clinical education; and presence of accredited post-professional education. Findings reveal that at least 11 of the professions studied offer first professional degrees. Differences were noted in duration and sequencing of undergraduate education, didactic or classroom education, and especially supervised practice. Models to facilitate comparison between educational standards were developed. The current educational model in dietetics is designed to prepare entry-level practitioners and academics who comprise less than 20% of the profession. This review supports the need to investigate educational opportunities for beyond-entry-level dietitians, and to develop educational programs that amplify the existing models for educating dietitians. PMID- 15746832 TI - n-3 long-chain polyunsaturated fatty acids in type 2 diabetes: a review. AB - Historically, epidemiologic studies have reported a lower prevalence of impaired glucose tolerance and type 2 diabetes in populations consuming large amounts of the n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFAs) found mainly in fish. Controlled clinical studies have shown that consumption of n-3 LC-PUFAs has cardioprotective effects in persons with type 2 diabetes without adverse effects on glucose control and insulin activity. Benefits include lower risk of primary cardiac arrest; reduced cardiovascular mortality, particularly sudden cardiac death; reduced triglyceride levels; increased high-density lipoprotein levels; improved endothelial function; reduced platelet aggregability; and lower blood pressure. These favorable effects outweigh the modest increase in low-density lipoprotein levels that may result from increased n-3 LC-PUFA intake. Preliminary evidence suggests increased consumption of n-3 LC-PUFAs with reduced intake of saturated fat may reduce the risk of conversion from impaired glucose tolerance to type 2 diabetes in overweight persons. Reported improvements in hemostasis, slower progression of artery narrowing, albuminuria, subclinical inflammation, oxidative stress, and obesity require additional confirmation. Expected health benefits and public health implications of consuming 1 to 2 g/day n-3 LC-PUFA as part of lifestyle modification in insulin resistance and type 2 diabetes are discussed. PMID- 15746833 TI - Obesity-related beliefs predict weight loss after an 8-week low-calorie diet. AB - The objective of this study was to examine whether beliefs about the cause, consequences, time line, and control of obesity are predictors of the amount of weight loss after an 8-week, low-calorie diet consisting of meal replacements. Forty-eight women and 18 men, mean age=45.9 (range=23 to 73 years) years and body mass index between 30 and 50 participated in a weight-loss program. Beliefs were measured at baseline by the Obesity Cognition Questionnaire and by an eating behavior self-efficacy scale. Correlational and regression analyses were performed to examine whether beliefs predicted weight change. Changes in body mass index, waist circumference, and blood pressure were significant (P <.001). Less weight reduction was associated with poor self-efficacy (r =-0.34, P <.01) and the beliefs that obesity had a physical origin (r =0.27, P =.04) and was not under behavioral control (r =-0.25, P =.04). Self-efficacy remained a significant predictor in regression analysis. The results suggest that the outcome of dietary interventions may be improved when adjusting beliefs, especially self-efficacy. PMID- 15746834 TI - College students can benefit by participating in a prepaid meal plan. AB - We compared Food Guide Pyramid (FGP) intake of college students according to their participation in a campus prepaid meal plan and their sex. Subjects were 503 single undergraduates (73 males, 430 females) who completed a 3-day dietary analysis assignment for an introductory nutrition course. Average intake for virtually all students fell short of FGP recommendations. We examined the difference between students' mean intake of foods from each FGP group and the recommended intakes. When compared with other student categories and food groups, male meal plan participants' vegetable and meat intakes were closer to recommended intakes, and meal plan participants' and females' fruit intakes were closer to recommended intakes. However, nonparticipants were closer than participants to recommended grain intakes. Participation in the prepaid campus meal plan appears to offer modest nutritional benefits to students through increased servings of foods from fruit, vegetable, and meat groups. PMID- 15746836 TI - Promoting healthful diets and exercise: efficacy of a 12-week after-school program in urban African Americans. AB - This study examined the effectiveness of a unique extracurricular after-school initiative designed to promote healthy diets and exercise in urban African Americans. The Students and Parents Actively Involved in Being Fit after-school program was offered for 12 weeks to students and their parents/guardians at an urban middle school. Specific aims of the intervention were to increase participants' vegetable and fruit intake by using established 5 A Day for Better Health educational resource materials/activities and to affect their health related fitness through dance, games, and fitness activities. Fifty-six children and 25 parents/guardians completed a standard battery of evaluations before and after the program. Pre-post pairwise t test revealed that both children and their parents/guardians showed an increase in fruit consumption and a reduction in diastolic blood pressure (P <.05). Moreover, children showed improvements in systolic blood pressure and fruit juice, salad, and nonfried potato consumption while parents/guardians showed a decrease in body fat, body mass index, and endurance walk/run time (P <.05). Overall, findings indicate that children tended to gain more diet-related benefits while parents/guardians tended to derive more fitness-related benefits. After-school programs like the Students and Parents Actively Involved in Being Fit initiative can potentially contribute to improved health levels in urban African Americans. PMID- 15746835 TI - Almonds in the diet simultaneously improve plasma alpha-tocopherol concentrations and reduce plasma lipids. AB - The objective of this study was to assess the dose-response effect of almond intake on plasma and red blood cell tocopherol concentrations in healthy adults enrolled in a randomized, crossover feeding trial. Participants were 16 healthy men and women, aged 41+/-13 years. After a 2-week run-in period, participants were fed three diets for 4 weeks each: a control diet, a low-almond diet, and a high-almond diet, in which almonds contributed 0%, 10%, and 20% of total energy, respectively. Changes in blood tocopherol levels were assayed by high pressure liquid chromatography. Incorporating almonds into the diet helped meet the revised Recommended Dietary Allowance of 15 mg/day alpha-tocopherol and increased lipid-adjusted plasma and red blood cell alpha-tocopherol concentrations. A significant dose-response effect was observed between percent energy in the diet from almonds and plasma ratio of alpha-tocopherol to total cholesterol. PMID- 15746837 TI - Practice paper of the American Dietetic Association: dietary supplements. PMID- 15746840 TI - What are acceptable cholesterol levels for children and adolescents? PMID- 15746843 TI - West Nile virus: a primer for the otolaryngologist. AB - BACKGROUND: Since recognition in the United States with a 1999 New York City epidemic, West Nile virus has enduringly migrated westward, leaving few states unaffected. Infection rates are rising at an alarming rate, doubling every year since introduction, with more than 9800 cases in 2003 alone and more than 260 deaths. Patients may present with myriad symptoms including a maculopapular rash that affects the face and trunk and diffuse lymphadenopathy, both of which may result in the initial consultation of the otolaryngologist. We review the clinical history of West Nile virus and its epidemiology, laboratory findings, and variable clinical presentation, with an emphasis on otolaryngologic manifestations. STUDY DESIGN AND SETTING COMPREHENSIVE: review of the literature over the past 50 years with an emphasis on what the present-day otolaryngologist needs to know concerning West Nile virus. Clinical manifestations of the head and neck such as encephalitis, meningitis, maculopapular rash, lymphadenopathy and dysphagia are discussed. RESULTS: To date, there are no articles in the otolaryngology literature discussing West Nile virus. These patients may present initially to multiple providers in diverse specialties because of multifarious initial signs and symptoms. The otolaryngologist must be educated on this quickly growing affliction and practice with a high index of suspicion. CONCLUSIONS: In this article we describe the clinical manifestations of West Nile virus, with an emphasis on the otolaryngologic manifestations. The otolaryngologist must become educated about this entity to facilitate preventative measures, adequately treat, and assist other providers in hopeful control and potential eradication of this infectious threat. PMID- 15746844 TI - Cat-scratch disease: Otolaryngologic manifestations and management. AB - OBJECTIVE: The bacteria Bartonella henselae has been known as the principal causative agent of cat-scratch disease (CSD) since 1992. It is an important cause of infectious lymphadenopathies in the head and neck. Nevertheless, CSD often remains unrecognized in cases of cervicofacial lymph node enlargement. STUDY DESIGN: Between January 1997 and May 2003, we conducted a prospective clinical study including 721 patients with primarily unclear masses in the head and neck. RESULTS: CSD was diagnosed by serology and molecular investigations in 99 patients (13.7%; median age 33 years). Cervicofacial lymphadenopathy was the most common manifestation. Atypical manifestation of CSD including Parinaud's oculoglandular syndrome, swelling of the parotid gland and erythema nodosum were diagnosed in 8.1%, 8.1%, and 2.0% of cases, respectively. CONCLUSIONS: Our results demonstrate that CSD is a major cause of enlarged cervicofacial lymph nodes and should therefore be included in the differential diagnosis of lymphadenopathy in the head and neck region. PMID- 15746845 TI - A systematic review of the diagnosis and treatment of primary hyperparathyroidism from 1995 to 2003. AB - OBJECTIVE: To systematically review the current preoperative diagnostic modalities, surgical treatments, and glandular pathologies associated with primary hyperparathyroidism. STUDY DESIGN: A systematic literature review. RESULTS: Of the 20,225 cases of primary hyperparathyroidism reported, solitary adenomas (SA), multiple gland hyperplasia disease (MGHD), double adenomas (DA), and parathyroid carcinomas (CAR) occurred in 88.90%, 5.74%, 4.14%, and 0.74% of cases respectively. Tc 99m -sestamibi and ultrasound were 88.44% and 78.55% sensitive, respectively, for SA, 44.46% and 34.86% for MGHD, and 29.95% and 16.20% for DA, respectively. Postoperative normocalcemia was achieved in 96.66%, 95.25%, and 97.69% of patients offered minimally invasive radio-guided parathyroidectomy (MIRP), unilateral, and bilateral neck exploration (BNE). Intraoperative PTH assays (IOPTH) were helpful in approximately 60% of bilateral neck exploration conversion (BNEC) surgeries. CONCLUSION: The overall prevalence of multiple gland disease (MGD and DA) was lower than often suggested by conventional wisdom. Furthermore, preoperative imaging was less accurate than it is often perceived for accurately imaging MGD. MIRP and UNE were more successful in achieving normocalcemia than is typically quoted. IOPTH was a helpful but not "fool-proof" adjunct in parathyroid exploration surgery. SIGNIFICANCE: These results support a greater role for the treatment of primary hyperparathyroidism using less invasive approaches. EMB rating: B-3. PMID- 15746846 TI - The utility of the temporalis muscle flap for oropharyngeal, base of tongue, and nasopharyngeal reconstruction. AB - OBJECTIVE: To determine the efficacy of temporalis muscle flap reconstruction of various defects of the oropharynx, nasopharynx, and base of tongue. STUDY DESIGN: Retrospective chart review of a consecutive series of 24 patients who underwent a total of 26 temporalis flaps (2 bilateral) by the senior author (Y.D.) from September 1997 to August 2003 for reconstruction of defects of the oropharynx, nasopharynx, and base of tongue. METHODS: Variables and outcomes that were examined included defect location, size, adjunctive therapy, complications, and ability to tolerate oral intake at follow-up. RESULTS: There was no evidence of flap failure in our series of patients. There were 2 cases of minor flap loss related to early prosthetic rehabilitation. Two cases of transient frontal nerve paralysis were noted. A 30.8% rate of complication (all minor) was noted in this study. At a mean follow-up of 12 months, 54.2% of patients were tolerating a full diet, 37.5% were tolerating most of their nutrition by mouth, and 8.3% were g tube dependent. CONCLUSION: The temporalis muscle flap represents an excellent alternative in reconstruction of otherwise difficult-to-reconstruct defects of the nasopharynx, oropharynx, and base of tongue. Donor site aesthetics are well accepted by patients with primary hydroxyapatite cement cranioplasty with or without secondary lipotransfer. PMID- 15746847 TI - The full-thickness forehead flap for complex nasal defects: a preliminary study. AB - OBJECTIVE: To report a new technique using a bivalved, full-thickness paramedian forehead flap. The unique vascular anatomy of the supratrochlear artery allows the skin and subcutaneous tissue to be separated from the frontalis muscle and pericranium. The deep layers serve as a pliable, vascularized intranasal lining. Bone and cartilage grafts can be placed as "sandwich" grafts between the deep and superficial layers of the flap. STUDY DESIGN: A retrospective review of 5 cases. RESULTS: All flaps survived. Four minor complications occurred in 3 patients. These resolved with minimal treatment. CONCLUSIONS: The full-thickness forehead flap is a viable option for large defects or for the difficult situation in which intranasal local flaps are not an option. SIGNIFICANCE: The gold standard for replacement of the intranasal lining is a septal mucosal or vestibular local flap. The full-thickness forehead flap is an option in patients for whom other lining flaps are not available. EBM RATING: C-3. PMID- 15746848 TI - Elective neck dissection versus observation in primary parotid carcinoma. AB - OBJECTIVE: To evaluate the efficacy of elective neck dissection in the clinically negative neck of patients with primary carcinoma of the parotid gland. Study design and setting A retrospective analysis was undertaken at a university Department of Otorhinolaryngology-Head and Neck Surgery on 83 previously untreated patients with primary carcinoma of the parotid gland and a clinically negative neck. The reliability of fine needle aspiration cytology, frozen section, and the clinico-pathologic findings of patients with occult neck metastases were analyzed. The regional recurrence rate and the outcome were compared among 2 groups; one with elective neck dissection (N = 41) and one without elective neck dissection (N = 42). RESULTS: The diagnosis of malignancy was known preoperatively in 59 (71%) cases, the exact histologic tumor type in 36 (43%) and the grade in 37 (44%) of 83 cases. Occult metastases were detected in 8 (20%) of 41 cNO patients, in 5 cases associated with a high-grade and in 3 cases with a low-grade carcinoma. Recurrence of disease developed in 5 (12%) patients in the elective neck dissection group and in 11 (26%) patients in the observation group. All of the 7 neck recurrences occurred in the observation group. The 5 year actuarial and disease-free survival rate was 80% and 86% for patients with elective neck dissection and 83% and 69% for patients without neck dissection. Conclusion and significance A routine elective neck dissection is suggested in all patients with primary carcinoma of the parotid gland. The efficacy of elective neck dissection, nevertheless, has never been evaluated prospectively. PMID- 15746849 TI - Incidence, age at onset, and potential reasons of malignant transformation in recurrent respiratory papillomatosis patients: 20 years experience. AB - Forty-two patients with recurrent respiratory papillomatosis (RRP) were accepted into a multicenter prospective study in 1983 to 1990, treated with alfa-IFN 3 MU/m 2 3 times a week and then followed-up until August 1, 2003. All the patients who had disease progression with pulmonary spread were characterized by insufficient response to IFN-therapy and detection of HPV type 11. Five patients (4/5 smokers) presented malignant transformation in lungs or nasopharynx (mean RRP duration was 27.2 +/- 8 years from RRP onset and 14.6 +/- 6.3 years from pulmonary spread until malignant transformation) with persistent RRP in larynx. The results of long-term follow-up in RRP patients with HPV 11 underline the necessity of reanalyzing the current therapy. PMID- 15746851 TI - Surgical management of parapharyngeal space tumors: a 10-year review. AB - OBJECTIVE: The purpose of this article is to describe the diagnostic evaluation and surgical approaches to parapharyngeal space tumors in a tertiary referral center. STUDY DESIGN AND SETTING: The study is a retrospective review of 47 patients diagnosed with tumors of the parapharyngeal space (12 with malignant diseases and 35 with benign lesions) and surgically treated during a 10-year period. The transcervical (40%) and the transcervical-transparotid approaches (46%) were the most commonly performed surgical procedures followed by the orbitozygomatic-middle fossa approach (12%) and the transmandibular approach (2%). RESULTS: The surgical procedures were uneventful and there were no postoperative mortalities. Complications were rare; the most common was transient facial nerve paralysis (5 patients). After an average follow-up of 35 months, only 1 of 35 patients with benign diseases had a recurrence 5 years following transcervical resection of a pleomorphic adenoma. Of 12 patients with malignant tumors, 5 (42%) are alive with no evidence of disease. The sensitivity of preoperative fine needle aspiration biopsy (n = 23 patients) was 87% for detection of malignant disease and specificity was 100%. CONCLUSIONS: Most benign parapharyngeal space tumors can be removed surgically with a low rate of complications and recurrence. Malignant neoplasms, however, carry an ominous prognosis and a low rate of disease-free survival. Fine needle aspiration may be helpful in preoperative diagnostic evaluation of patients with parapharyngeal space tumors. PMID- 15746850 TI - Overexpression of MMP-2 in laryngeal squamous cell carcinoma: a potential indicator for poor prognosis. AB - OBJECTIVES: To investigate the expression and clinical significance of gelatinases (MMP-2 and MMP-9) in patients with laryngeal squamous cell carcinoma (LSCC). STUDY DESIGN AND SETTING: In a retrospective study of 72 consecutive patients with LSCC hospitalized in a single cancer center, immunohistochemistry was used to examine the expression of MMP-2 and MMP-9 in surgical samples. The results were compared to clinicopathological features and prognosis. RESULTS: The positive expression of MMP-2 and MMP-9 in patients with LSCC was 50% (36/72) and 73.6% (53/72), respectively. According to the expression scale, there were 36 patients of -, 26 patients of +, 7 patients of ++, and 3 patients of +++ expression of MMP-2; 19 patients of -, 26 patients of +, 16 patients of ++, and 11 patients of +++ expression of MMP-9. There was no significant relationship found between the expression of MMP-2 or MMP-9 and clinicopathological features of LSCC, such as histological grade, primary site, T stage, N stage, and clinical stage. The 5-year overall survival (OS) and disease-free survival (DFS) rate calculated by Kaplan-Meier method in patients with negative and positive expression of MMP-9 and MMP-2 was 73.68%, 50.94%, 73.68%, and 49.06% in MMP-9 and 72.22%, 41.67%, 72.22%, and 38.89% in MMP-2, respectively. Significant 5-year survival difference was found between patients with negative and positive expression of MMP-2 (log rank = 6.74, P = 0.0094). There was significant lower survival rate in patients with higher positive expression of MMP-2 (log rank = 11.77, P = 0.0028). In glottic laryngeal cancer, positive expression of MMP-2 could predict poor survival and was more likely to present primary recurrence. CONCLUSION: The expression of MMP-2 could be used as a potential predictor for poor prognosis in patients with LSCC. PMID- 15746852 TI - Routine histopathology in uncomplicated sinus surgery: is it necessary? AB - OBJECTIVE: To evaluate the incidence of occult pathology in routine, uncomplicated endoscopic sinus surgery, and to suggest guidelines for when to send specimens for histopathologic exam. STUDY DESIGN AND SETTING: Retrospective analysis of case records of 790 patients who underwent 868 endoscopic sinus surgeries at a tertiary care center from 1986 to 2003. Indications were chronic sinusitis, recurrent acute sinusitis, nasal polyposis, or combinations of these diagnoses. All cases were considered routine and did not involve preoperative suspicion of neoplasm or other complicating factors. Charts were reviewed for surgical indication, patient age, laterality of disease, history of prior sinus surgery, intraoperative suspicion of tumor, and final histopathology. RESULTS: In 868 cases of endoscopic sinus surgery, occult neoplasm was diagnosed in 2 patients (0.23%). In one patient, the initial surgery cured the lesion. Final histopathology of the remaining 866 (99.8%) specimens was consistent with inflammation and/or nasal polyposis. In 121 cases of unilateral sinusitis, none was positive for neoplasm. In 277 cases involving bilateral nasal polyposis and 13 involving unilateral polyposis, no neoplasms were found. Intraoperative suspicion of neoplasm occurred in 12 cases, with all specimens read as consistent with sinusitis and/or polyposis. CONCLUSIONS: Histopathologic review of every specimen obtained in routine sinus surgery for sinusitis and/or nasal polyposis is not indicated. Submission of specimen is indicated in routine cases when: 1) there is intraoperative suspicion of tumor, 2) unilateral nasal polyposis is present, 3) unilateral sinus opacification is present, and 4) additional diagnostic information is needed (eg, presence of eosinophils, fungal forms, etc.). SIGNIFICANCE: Establishes a safe and reasonable standard of care, with potential cost savings and medico-legal ramifications. PMID- 15746854 TI - Chronic rhinosinusitis and biofilms. AB - BACKGROUND AND HYPOTHESIS: Biofilms have been implicated in several head and neck infectious processes such as the following: dental and periodontal disease, otitis media, tympanostomy tube otorrhea, and chronic tonsillitis. We believe that biofilms also are associated with chronic rhinosinusitis. No information is known regarding the presence of biofilms in chronic rhinosinusitis. STUDY DESIGN AND SETTING: With institutional review board approval, tissue was obtained from consenting chronic rhinosinusitis patients who were undergoing functional endoscopic sinus surgery. Specimens were taken bilaterally from the ethmoid and maxillary sinuses. Inclusion criteria consisted of a positive diagnosis with pathologic tissue confirmation of chronic inflammation. Diagnosis was based on patient history, physical exam, and coronal sinus CT findings. Once collected, the specimens were labeled and fixed in formalin. The specimens were subsequently dehydrated, with successive immersions in increasing concentrations of diluted ethanol. The specimens were allowed to air dry and then were affixed to aluminum stubs with colloidal carbon. The sample surface was coated with a gold and palladium layer. The specimens were examined under an electron microscope. Areas of interest were photographed. RESULTS: Specimens from 5 patients were examined. All revealed bacterial biofilms. Invariably, biofilms were seen in the ethmoid, as well as in other samples. Denudation of ciliated and goblet cells was noted in all specimens. Biofilms resembled that of Staphylococcus species. Unidentified biofilms were also seen. CONCLUSIONS: This is the first documentation of biofilms in association with chronic rhinosinusitis. Further investigation is warranted, especially with control research subjects. PMID- 15746855 TI - Endoscopic management of cerebrospinal fluid leak associated with the use of bone wax in skull-base surgery. AB - OBJECTIVE: To alert otolaryngologists to consider the possibility that bone wax may be associated with cerebrospinal fluid leaks that occur immediately after skull-base craniotomy approaches. STUDY DESIGN AND SETTING: Clinical report from surgical experience in a tertiary care setting. RESULTS: Three patients presented with brisk cerebrospinal fluid leak after craniotomy. Sinus endoscopy revealed bone wax within a small parasphenoid defect in all 3 cases. CONCLUSIONS: Bleeding from areas of the skull base adjacent to the paranasal sinuses during craniotomy can signal a small breach of the skull base. Bone wax may control bleeding from edges of transected bone but also stent the skull-base defect open, preventing fibrin deposition and spontaneous healing and closure. SIGNIFICANCE: Otolaryngologists repairing a cerebrospinal fluid leak after a skull-base craniotomy approach should consider the possibility of encountering bone wax and be able to identify it to better treat their patient. PMID- 15746856 TI - The accuracy of computer-aided surgery in neurotologic approaches to the temporal bone: a cadaver study. AB - OBJECTIVE: To assess the accuracy of computer-aided surgery for common neurotologic approaches to the temporal bone. STUDY DESIGN AND SETTING: Cadaveric heads were dissected by using standard neurotologic approaches to the temporal bone including translabyrinthine, middle fossa, and retrosigmoid. Dissected anatomic landmarks from each approach were compared with CT images that were obtained before dissection on the VectorVision 2 system (BrainLAB Corp, Munich, Germany) and the variability measured from digital images. Each approach was performed 5 times, with each anatomic landmark measured 3 times from each approach. RESULTS: The accuracy of the computer-aided surgery system was less than 1 mm for all anatomic points measured. Forty-two of the 49 measured points were accurate to less than 0.5 mm. CONCLUSIONS: Computer-aided surgery of the temporal bone using common neurotologic approaches can be performed accurately and reliably in cadaver specimens. SIGNIFICANCE: The utility and limitations of computer-aided surgery in the temporal bone are discussed. EBM RATING: B-2. PMID- 15746857 TI - The analysis of maxillary sinus aeration according to aging process; volume assessment by 3-dimensional reconstruction by high-resolutional CT scanning. AB - OBJECTIVE: To evaluate change of the maxillary sinus volume according to patient age and gender by using a 3-dimensional (3-D) reconstruction of computed tomography images. STUDY DESIGN AND SETTING: One hundred seventy-three people (totaling 238 maxillary sinuses) who had undergone paranasal sinus CT scan between December 2000 and November 2003 and had no evidence of inflammation or hypoplasia in the CT finding and had no specific history of paranasal sinus surgery or maxillofacial trauma were retrospectively analyzed. The 3-D reconstruction images were obtained by using a surface-rendering technique (Vworks; CybeMed, Seoul, Korea) on a personal computer. The mean volume of maxillary sinus was evaluated according to patient chronologic age and gender. The ratio of the maximum horizontal and half-horizontal extension for the estimation of the morphological change of maxillary sinus and the degree of descent of the sinus below the nasal floor were evaluated in the 3-D image. RESULTS: The development of the maxillary sinus continued until the 3rd decade in males and until the 2nd decade in females. The mean maxillary sinus volume in early adults was 24,043 mm 3 (males) and 15,859.5 mm 3 (females). There was a significance difference in the sinus volume ( P < 0.05) according to gender, and there was a significant difference in the maxillary sinus volume according to age before it reached maximum. After its maximum growth period, however, there was no significant difference in the volume change of maxillary sinus and the descent below the nasal floor between two adjacent groups. CONCLUSIONS: The growth of the maxillary sinus continues until the 3rd decade in males and the 2nd decade in females. Therefore, a maxillary sinus operation affecting the bony structures before these ages might affect the development of the sinus and needs to be performed carefully. PMID- 15746858 TI - In vitro assessment of image-guided otologic surgery: submillimeter accuracy within the region of the temporal bone. AB - OBJECTIVES: Application of image-guided surgery to otology has been limited by the need for submillimeter accuracy via a fiducial system that is easily usable (noninvasive and nonobstructive). METHODS: A dental bite-block was fitted with a rigid frame with 7 fiducial markers surrounding each external ear. The temporal bones of 3 cadaveric skulls were removed and replaced with surgical targets arranged in a bull's-eye pattern about the centroid of each temporal bone. The surgical targets were identified both within CT scans and in physical space using an infrared optical tracking system. The difference between positions in CT space versus physical space was calculated as target registration error. RESULTS: A total of 234 independent target registration errors were calculated. Mean +/- standard deviation = 0.73 mm +/- 0.25 mm. CONCLUSIONS: These findings show that image-guided otologic surgery with submillimeter accuracy is achievable with a minimally invasive fiducial frame. Significance In vivo validation of the system is ongoing. With such validation, this system may facilitate clinically applicable image-guided otologic surgery. EBM RATING: A. PMID- 15746859 TI - Surgical management of Meniere's disease in the era of gentamicin. AB - OBJECTIVE: For many years, surgery was the mainstay of therapy for medically refractory patients, but recently, transtympanic gentamicin perfusion has attracted increasing interest and is a method frequently used for treating Meniere's disease. Many otologists question the relevance of surgical treatments, and traditional options are rarely discussed or offered to patients. The purpose of this study is to describe results of labyrinthectomy, vestibular nerve section, and endolymphatic mastoid shunt surgery for patients with Meniere's disease and to compare them with published results for gentamicin perfusion. STUDY DESIGN AND SETTING: Retrospective chart review. Two hundred twenty-nine patients underwent surgery for management of Meniere's disease between January 1, 1995 and December 31, 2001. One hundred eighty-nine patients' charts had sufficient data for review. Thirty-two patients had translabyrinthine labyrinthectomies, 83 underwent suboccipital vestibular nerve sections, and 74 elected for an endolymphatic mastoid shunt. Hearing results, dizziness classification, and functional level score were determined from patient charts and telephone conversations. All results were in accordance with the guidelines of the AAO-HNS Committee on Hearing and Equilibrium for evaluation of Meniere's disease therapy. RESULTS: Audiologic results, functional level score, and dizziness classification are reported for the preoperative period and for the 18- to 24-month postoperative period for all surgical patients. These data are also reported individually for each of the 3 surgical procedures. Early postoperative data and most recent follow-up data are presented if available. CONCLUSIONS: Surgical management of Meniere's disease is a safe and viable option for patients with medically refractory disease. EBM RATING: C. PMID- 15746860 TI - Can mobile phone emissions affect auditory functions of cochlea or brain stem? AB - PROBLEMS ADDRESSED: Despite their abundant spread, mobile phones are suspected by a major share of the population to cause adverse effects on health and welfare. The ear as the sense organ next to the individual device has rarely been investigated for short-term effects in this regard. In a previous article, we could not prove any impact on the vestibular part of the inner ear. Our present examinations are concerned with the question whether mobile phone emissions could affect cochlear or auditory brain stem functions. METHODS AND MEASURES: In 12 healthy test persons with normal hearing, auditory brain stem reflexes recordings were performed before, during, and after exposure to electromagnetic emissions by standardized mobile phone devices. Two modes of electromagnetic emissions fields were administered: pulsed and continuous. For acoustic stimulation simultaneous to field exposure, special "plug-in" earphones had to be used. RESULTS: No impact on auditory brain stem reflexes recordings in terms of absolute and interpeak latencies could be found. CLINICAL SIGNIFICANCE: Together with the results of a previous article concerned with the vestibular part of the inner ear, we can state that there are no adverse effects of mobile phone emissions on the ear function, at least on a short-term range. Of course, any long-term effects cannot be excluded by our study. PMID- 15746861 TI - Long-term results of cochlear implantation in children. AB - OBJECTIVES: Since the first implantation of a multichannel cochlear implant over 15 years ago there have been improvements in implant devices, surgical techniques, speech coding strategies and rehabilitation matters. There also have been concerns, whether the first implanted systems are stable and the benefit for the patients is constant over a long time. This study analyzes long-term results of prelingually deaf children who were implanted at Aachen University Hospital. STUDY DESIGN AND SETTING: We examined 16 prelingually deaf children, who underwent cochlear implantation at our department between 1990 and 1993 over a time period of 10 to 13 years. The indication, perioperative complications, technical parameters, speech test results as well as the psychosocial development of the children were analyzed. A questionnaire containing questions about the daily use, expectations and the personal evaluation of the procedure was sent to all patients. RESULTS: The mean age of all patients was 9 years at time of implantation. In two cases we saw postoperative complications which needed an intervention. The speech test results remained stable after reaching a plateau after 5 years. Eighty-eight percent of all patients would undergo the procedure again and 94% would recommend cochlear implantation to others. CONCLUSION: Cochlear implantation in cases of prelingual deafness is feasible without severe complications and leads to an increasing quality of life demonstrated by long term observation. PMID- 15746862 TI - Is the middle fossa approach the treatment of choice for intracanalicular vestibular schwannoma? AB - OBJECTIVES: To compare the 2 surgical techniques most commonly used during vestibular schwannoma (VS) surgery, i.e., the middle fossa (MF) and the retrosigmoid-transmeatal (RS-TM) routes, when hearing preservation is attempted. STUDY DESIGN: A longitudinal study of a series of consecutive patients operated on with the 2 techniques by the same surgeon was conducted. Selection criteria included tumor confined to the internal auditory canal (IAC) with a length ranging from 4 to 12 mm and hearing class A or B. Patients were alternately assigned to 1 of the 2 groups regardless of auditory class and distance of the tumor from the IAC fundus. Thirty-five subjects were operated on with the RS-TM technique and 35 via the MF route. RESULTS: No significant differences in auditory and facial nerve function results between the 2 techniques were observed. The RS-TM approach, however, showed better facial nerve results at discharge. VS size, IAC enlargement, and, particularly, the distance from the IAC fundus were found to influence the postoperative results more than the type of approach itself. CONCLUSIONS: The MF approach has been described as being the better technique for VS surgery in terms of auditory results. However, this claim lacks statistical substantiation because no prospective studies are to be found in the literature. The present longitudinal investigation shows that the MF approach does not afford any particular advantages over the RS-TM route in terms of auditory results in intracanalicular VS, with the exception of tumors reaching the IAC fundus. PMID- 15746863 TI - Paragangliomas in patients with mutations of the SDHD gene. AB - OBJECTIVE: Paragangliomas represent neoplasms of neural crest origin that arise from paraganglia. Mutations in the gene encoding succinate dehydrogenase subunit D (SDHD) are responsible for a percentage of hereditary paragangliomas. We previously described a group of 271 pheochromocytoma patients, 11 of whom had mutations of the SDHD gene. The objective of this study was to find out whether those 11 patients had additional paragangliomas. STUDY DESIGN: Ten patients participated in our clinical screening program that included MRI of the skull base and neck, thorax, and abdomen, as well as an 18 Fluoro-DOPA positron emission tomography (DOPA-PET). RESULTS: Five patients presented with head and neck paragangliomas, 1 patient with a thoracic paraganglioma, and 2 patients with intraabdominal paragangliomas. CONCLUSIONS: The screening for paragangliomas in patients with mutations of the SDHD gene offers the chance to diagnose those tumors in an early stage. SIGNIFICANCE: Because morbidity after surgical resection increases with tumor size, early surgery will minimize the potential risks. PMID- 15746864 TI - A novel laryngoscope instrument stabilizer for operative microlaryngoscopy. AB - OBJECTIVE: To evaluate and optimize the design of a removable and inexpensive internal stabilization device to reduce the effect of intention tremor during laryngeal microsurgery. STUDY DESIGN AND SETTING: In this laboratory investigation, stabilizers were designed and constructed to allow a nonobstructing view of the surgical field, permit simple insertion and removal, and accommodate microsurgical instruments. Prototype stabilizers were tested by using a Dedo laryngoscope, a measurement grid, and video recording equipment, which recorded instrument tremor within the magnified operative field for later analysis. Physicians also rated instrument stability, mobility, visualization, and ease of use on a survey form. RESULTS: Instrument tremor was reduced approximately 90%, with little obstruction of view of the surgical field. Instrument range of motion was reduced but improved rapidly as the stabilizer bar was moved further from the tip of the laryngoscope. CONCLUSIONS: Use of a stabilization device in the laryngoscope lumen reduces instrument tremor and has the potential to improve surgical performance during laryngeal microsurgery. EBM RATING: B-3. PMID- 15746865 TI - Vestibular fold flap for post-cordectomy laryngeal reconstruction. AB - OBJECTIVES: The development of techniques causing fewer sequelae without compromising oncologic control has led to better phonation, breathing, and postoperative swallowing in patients submitted to cordectomy. The objective of the present study was to describe post-cordectomy laryngeal reconstruction with a vestibular fold flap and to analyze the resulting laryngeal functions. STUDY DESIGN: Prospective study. METHODS: Ten patients, 9 males and 1 female, aged 45 to 75 years (mean, 64.5 years), were evaluated. The patients had squamous cell carcinomas in the glottic region and were treated surgically by laryngofissure and cordectomy with the use of a vestibular fold flap for laryngeal reconstruction. The patients were followed up for functional laryngeal analysis that consisted of videolaryngostroboscopy, endoscopic evaluation of swallowing, and voice recording for perceptive auditory analysis, and for computing purposes. RESULTS: No patients depended on tracheostomy during the late postoperative period. No stenosis, anterior synechia, granuloma, or laryngocele were detected, although 1 patient developed immobility in the median position of the operated hemilarynx, which, however, was not sufficient to impair the respiratory function. No alterations in the pharyngeal phase of swallowing were observed by endoscopic evaluation. Using the GRBAS scale, we observed moderate (30%) and severe (20%) dysphonia in 50% of the patients and grade 1 dysphonia (40%) or normal (10%) dysphonia in the remaining ones. Computing analysis revealed the following mean values: fundamental frequency of 177.5 Hz, jitter 1.11%, and shimmer 7.04%. CONCLUSIONS: We conclude that reconstruction with a vestibular fold flap permitted the maintenance of the laryngeal functions of breathing and airway protection during swallowing, as well as the maintenance of phonation function, providing perfect voice emission according to perceptive auditory or acoustic analysis in 1 patient and moderate or severe dysphonia in one half of the cases when technical faults occurred. PMID- 15746866 TI - The influence of tracheotomy tubes on the swallowing frequency in neurogenic dysphagia. AB - OBJECTIVES: To compare the swallowing frequency in patients with neurogenic dysphagia with or without tracheotomy tubes (TT) to assess the underlying mechanisms of dysphagia to improve rehabilitation strategies. STUDY DESIGN AND SETTING: Prospective study, 10 patients (64 +/- 7 years) with neurogenic dysphagia. Glasgow Coma Scale (GCS) less than 8 points, tracheotomy due to the dysphagia 2 weeks before the examination. The swallowing frequency (1 or less over 5 min) was assessed over 5 consecutive days with or without TT. RESULTS: The swallowing frequency increased after removal of the TT. These findings did not influence the GCS or the Coma Remission Scale. Over a 5-day period, the frequency of swallowing was increased. CONCLUSION: TTs decisively influence the swallowing behavior of vegetative patients. This phenomenon could be based on an improved sensitivity under re-established physiological expiration. We strongly favor removing the TT or deflating the cuff of the TT under therapeutic conditions in a rehabilitation therapy setting. PMID- 15746867 TI - The use of Ligasure Vessel Sealing System in thyroid surgery. AB - OBJECTIVES: To evaluate the role of Ligasure Vessel Sealing System (LVSS) (Valleylab, Boulder, CO) in thyroid surgery, with emphasis given to the duration of the procedure, as well as potential relevant postoperative complications. STUDY DESIGN AND SETTINGS: A prospective study was conducted from November 1, 2002 to October 31, 2003, on patients undergoing thyroid surgery with the use of LVSS device as the primary means of ligation. All patients' records were placed in a database. Efficacy of hemostasis, operation time, and postoperative complications were assessed. RESULTS: were compared with previous surgical procedures done by our team when the LVSS was not available. RESULTS: LVSS proved effective in providing ligation and hemostasis during thyroid surgery in 72 consecutive patients. There was a mean reduction in operating time of 23 minutes compared with previous surgical thyroid procedures. Transient hypocalcaemia occurred in 4 subjects (5.55%); one incidence of transient paresis of the recurrent laryngeal nerve was also observed. CONCLUSION: LVSS proved quite a reliable and safe device in thyroid surgery; it provided sufficient hemostasis and reducing operative time. PMID- 15746868 TI - Surgical complications after thyroid surgery performed in a cancer hospital. AB - OBJECTIVE: This study evaluates the incidence and risk factors of complications in patients submitted to thyroidectomy in a cancer hospital with residency training. STUDY DESIGN: A retrospective chart and complications review of 1020 patients (1990-2000) underwent to thyroidectomy. RESULTS: At our cancer hospital, 1020 patients underwent thyroidectomy. The main postoperative complications consisted of transient hypocalcemia in 134 (13.1%) patients, permanent hypocalcemia in 26 (2.5%) patients, transient vocal cord palsy in 14 (1.4%) patients, and permanent vocal cord palsy in 4 (0.4%) patients. The type of thyroidectomy, neck dissection, and paratracheal lymph node dissection were significantly associated with transitory and permanent hypocalcemia. CONCLUSION: Thyroid surgery can be performed safely in a surgical residency training program under direct supervision of an experienced surgeon with little morbidity to the patients. Hypocalcemia is the most significant complication. Neck and paratracheal lymph node dissections were the most significant predictors of hypocalcemia in patients who underwent total thyroidectomy. PMID- 15746869 TI - Reading performance in children with otitis media. AB - OBJECTIVE: To examine whether middle ear diseases and the associated hearing loss in early childhood affect reading performance later at school. STUDY DESIGN AND SETTING: One hundred and sixty children, 6.5 to 8 years of age, were enrolled in this study: 80 children with a history of recurrent infections and/or prolonged periods of effusions of the middle ear before the age of 5 years, and 80 healthy children without any history of middle ear disease. Data were collected from the medical records of the children. Every child underwent a complete otological and audiological evaluation, followed by special reading tests. RESULTS: The study group performed more poorly, in all reading tests, as compared to the controls ( P < 0.001). CONCLUSION: Children with recurrent or prolonged middle ear diseases during the first five years of life tend to be at greater risk for delayed reading than aged-matched controls with no previous middle ear diseases. PMID- 15746870 TI - Otolaryngologic presentations of amyotrophic lateralsclerosis. AB - OBJECTIVES/HYPOTHESIS: To determine the incidence of voice disturbance as a presenting symptom of amyotrophic lateral sclerosis (ALS) and describe laryngologic features of ALS. STUDY DESIGN: Retrospective review. METHODS: Records of patients with voice disturbance at a voice center and ALS patients at a neurology clinic were reviewed from January 1998 to March 2003. RESULTS: 15 of 1759 patients with voice disturbance were later diagnosed with ALS. Of 220 ALS patients presenting to neurology clinic, 44 had bulbar symptoms and 19 had initially presented to an otolaryngologist. Dysarthria, dysphagia, tongue fasciculation, and incomplete vocal fold closure were common findings. Neuromuscular disease was missed in 8 of 19 ALS patients seen by an otolaryngologist. CONCLUSIONS: Although otolaryngologists rarely encounter undiagnosed ALS patients, a significant portion of bulbar ALS patients are initially evaluated by otolaryngologists. SIGNIFICANCE: Vigilance for neuromuscular abnormalities on otolaryngologic exam is important in patients who present with dysarthria, dysphonia, or dysphagia. EBM RATING: C. PMID- 15746871 TI - Melanoma metastasis masquerading as bilateral acoustic neuromas. PMID- 15746872 TI - An intracranial vagal schwannoma without jugular foramen erosion or vagal dysfunction. PMID- 15746873 TI - Facial nerve schwannoma presenting as conductive hearing loss. PMID- 15746874 TI - Lipoblastoma of the neck mimicking cystic hygroma. PMID- 15746875 TI - Histoplasmosis presenting as upper airway obstruction. PMID- 15746876 TI - Idiopathic submandibular sialoceles in the neck. AB - A submandibular sialocele is a subcutaneous cavity containing saliva. The clinical and radiologic features of 3 patients with an idiopathic submandibular sialocele are presented. All 3 patients were males in their twenties. Submandibular sialocele presents as a soft cystic and compressible neck mass, with no history of previous trauma or diseases of the salivary gland. Computed tomography (CT) of the neck revealed a homogenous lesion with enhancing rim. The lesion appeared to be insinuating into the surrounding tissue. Excision of the sialocele, leaving the submandibular gland intact, was performed for the first patient. Recurrence of a neck mass occurred after 4 months. Complete excision of the sialocele with associated submandibular gland was subsequently performed. There was no recurrence after a follow-up period of 3 years. Excision of gland and sialocele was performed for the other 2 patients. There was no recurrence after a follow-up of 2 years and 10 months, respectively. PMID- 15746877 TI - Laryngeal paraganglioma in a patient with multiple head and neck paragangliomas. AB - Paragangliomas of the larynx are rare neuroendocrine tumors, with fewer than 70 cases reported in the literature. 1 Typically, laryngeal paragangliomas are not found in patients with multicentric or familial paragangliomas. Only 1 case of laryngeal paraganglioma has been reported in a patient with a synchronous lesion elsewhere (carotid body tumor). 2 We report an additional case of a patient with a laryngeal and multiple other paragangliomas. PMID- 15746879 TI - Preface to outcome measures in allergic rhinitis. PMID- 15746880 TI - Clinical outcomes and adverse effect monitoring in allergic rhinitis. AB - The subjective recording in diary cards of symptoms of itch, sneeze, nose running, and blockage, with the use of a rating scale to indicate the level of severity, is usual for clinical trials in allergic rhinitis. The primary outcome measure is usually a composite score that enables a single total symptoms score endpoint. It is appreciated, however, that rhinitis has a greater effect on the individual than is reflected purely by the recording of anterior nasal symptoms. Nasal obstruction is troublesome and may lead to sleep disturbance in addition to impaired daytime concentration and daytime sleepiness. These impairments affect school and work performance. Individuals with rhinitis find it socially embarrassing to be seen sneezing, sniffing, or blowing their nose. To capture these and other aspects of the disease-specific health-related quality of life, questionnaires such as the Rhinoconjunctivitis Quality of Life Questionnaire have been developed and validated for clinical trial use. The adoption of health related quality of life questionnaires into clinical trials broadens the information obtained regarding the effect of the therapeutic intervention and helps focus on issues relevant to the individual patient. It must be appreciated that it is not only the disease that may adversely affect health-related quality of life; administered therapy, although intended to be beneficial, may also cause health impairment. Adverse-event monitoring is thus essential in clinical trials. The first-generation H 1 -histamines, because of their effect on central H 1 receptors, are classically associated with central nervous system (CNS) effects such as sedation. Although this is not always perceived by the patient, it is clearly evident with objective performance testing, and positron emission tomography scanning has directly demonstrated the central H 1 -receptor occupancy. The second-generation H 1 -antihistamines have reduced central H 1 receptor occupancy and considerably reduced or absent CNS sedative effects. Therefore, the CNS effects are entirely avoidable, and the first-generation H 1 antihistamines should no longer be used in the management of allergic rhinitis. The considerably rarer but potentially very serious cardiac arrhythmogenic effects of H 1 -antihistamines are appreciated to be molecule-specific rather than class-specific. The in vitro screening of new compounds to eliminate the further development of those with cardiotoxicity ideally will lead to this adverse effect being historic. The incorporation of electrocardiogram recording in clinical trials provides direct information relating to prolongation of QT interval corrected for heart rate. Although administered at low doses, intranasal steroids still have the potential for systemic absorption and adverse consequences. However, it is appreciated that meaningful differences exist in the bioavailability of different steroid molecules, and although a small but statistically significant effect on growth in children has been identified with the long-term use of intranasal beclomethasone when administered twice daily for 1 year, this is not evident with all intranasal steroids. In addition, twice daily intranasal steroid administration may have more effect--from the endocrinologic perspective--than once-daily administration in the morning, which coincides better with the natural diurnal variation in cortisol. Thus, once-daily intranasal steroid administration is preferable, and when used in studies in children, measurement of height change during the study period is an important outcome variable together with other indices of systemic steroid bioavailability (eg, tests of hypothalamic-pituitary-adrenal axis function). These considerations have even greater relevance if children are concurrently also receiving inhaled steroids for asthma, because the total steroid load will be greater. PMID- 15746881 TI - Objective monitoring of nasal airway inflammation in rhinitis. AB - Allergic rhinitis is an inflammatory nasal disorder in which a range of different cells participates. A variety of approaches has been used to monitor nasal inflammation objectively to investigate disease processes and to evaluate the effect of therapeutic intervention. These approaches include nasal lavage, nasal cytology, and nasal biopsy, together with the more recently established measurement of nasal nitric oxide (NO) concentration. Although all provide information about nasal mucosal inflammation, the extent of information that can be obtained by each approach, the ease of sampling, and the complexity of sample handling differ. Such considerations influence the choice of approach when measurement of nasal inflammation is to be an objective outcome parameter in a clinical trial. In addition, the choice of approach is also determined by the questions or hypotheses that are to be addressed. Nasal lavage is simple and rapid to perform, is well tolerated, and provides a sample that can provide information about luminal cell recruitment, cell activation, and plasma protein extravasation. Nasal cytology involves sampling and recovering mucosal surface cells. It is also easy to perform and is well tolerated in general, although some find that the procedure causes a transient unpleasant sensation. A differential cell count from the sample provides information about relative cell populations. Both nasal lavage and nasal cytology are readily applicable to clinical trials. Nasal cytology sample handling is easier, but nasal lavage offers the advantage of providing considerably greater information from the sample. Nasal biopsy is a considerably more invasive procedure and requires expertise not only in tissue sampling but also in biopsy processing. Therefore, it is applicable only in specialist centers. However, nasal biopsy is the only sampling technique that directly informs about tissue cellular events, although these may be implied, in part from the other sampling approaches. Tissue specimens can be used to evaluate both protein and gene expression. Measurement of nasal NO involves expensive equipment but provides an instantaneous result, unlike the other approaches, all of which require sample processing and analysis. Recommendations for standardization of measurement have been made, and measures are considered in part to reflect allergic inflammation within the nasal mucosa. The limitations of nasal NO are that it reflects only a certain aspect of allergic mucosal inflammation, and that because a proportion of nasally measured NO is derived from the sinuses under normal circumstances, nasal NO is not specific for nasal disease. The high contribution from the sinus mucosa limits the discriminatory ability of nasal NO to reflect nasal tissue-specific alterations. The incorporation of measures of nasal inflammation in clinical trials has distinguished anti-inflammatory therapy from symptomatic therapy and has the potential to provide information about the efficacy of novel therapies for allergic rhinitis. PMID- 15746882 TI - Objective monitoring of nasal patency and nasal physiology in rhinitis. AB - Nasal obstruction can be monitored objectively by measurement of nasal airflow, as evaluated by nasal peak flow, or as airways resistance/conductance as evaluated by rhinomanometry. Peak flow can be measured during inspiration or expiration. Of these measurements, nasal inspiratory peak flow is the best validated technique for home monitoring in clinical trials. The equipment is portable, relatively inexpensive, and simple to use. One disadvantage, however, is that nasal inspiratory peak flow is influenced by lower airway as well as upper airway function. Rhinomanometry is a more sensitive technique that is specific for nasal measurements. The equipment, however, requires an operator, is more expensive, and is not portable. Thus, it is applicable only for clinic visit measures in clinical trials. Measurements require patient cooperation and coordination, and not all can achieve repeatable results. Thus, this objective measure is best suited to laboratory challenge studies involving smaller numbers of selected volunteers. A nonphysiological measure of nasal patency is acoustic rhinometry. This sonic echo technique measures internal nasal luminal volume and the minimum cross-sectional area. The derivation of these measures from the reflected sound waves requires complex mathematical transformation and makes several theoretical assumptions. Despite this, however, such measures correlate well with the nasal physiological measures, and the nasal volume measures have been shown to relate well to results obtained by imaging techniques such as computed tomography scanning or magnetic resonance imaging. Like rhinomanometry, acoustic rhinometry is not suitable for home monitoring and can be applied only to clinic visit measures or for laboratory nasal challenge monitoring. It has advantages in being easy to use, in requiring little patient cooperation, and in providing repeatable results. In addition to nasal obstruction, allergic rhinitis is recognized to be associated with impaired mucociliary clearance and altered nasal responsiveness. Measures exist for the monitoring of these aspects of nasal dysfunction. Although measures of mucociliary clearance are simple to perform, they have a poor record of reproducibility. Their incorporation into clinical trials is thus questionable, although positive outcomes from therapeutic intervention have been reported. Measures of nasal responsiveness are at present largely confined to research studies investigating disease mechanisms in allergic and nonallergic rhinitis. The techniques are insufficiently standardized to be applied to multicenter clinical trials but could be used in limited-center studies to gain insight into the regulatory effects of different therapeutic modalities. PMID- 15746883 TI - Clinical trial design, nasal allergen challenge models, and considerations of relevance to pediatrics, nasal polyposis, and different classes of medication. AB - Clinical trials in allergic rhinitis present several specific difficulties. In seasonal pollen-related disease, there are variations between subjects in the extent of pollen sensitization, individual variations in exposure to pollen even within a set area because of lifestyle differences, and variations between different areas in pollen counts and weather patterns. Thus, large patient numbers are needed in multicenter trials to account for such variations when the standard endpoint is symptom reporting. Furthermore, a pollen season may be relatively short (eg, lasting 6-8 weeks), and the pollen count is inconsistent during this period. Crossover study designs are thus inappropriate, and trials are usually conducted with a parallel-group design. This further increases the trial sample size as it reduces statistical power. These large patient numbers must be recruited over a very short period. Perennial house dust mite-sensitive allergic rhinitis presents other problems. Although there is less disease variation, it is appreciated that symptoms may be induced by nonallergic as well as allergic mechanisms because of the nasal hyperresponsiveness. The nonallergic symptoms may not be modified by treatments based on allergic disease mechanisms. Thus, symptom outcomes--although relevant to the patient--may not adequately reflect the pharmacologic efficacy of the specific intervention. To control variability and focus on allergic disease mechanisms, nasal allergen challenge has been used in drug development. Single-dose challenges in the laboratory or in a pollen chamber, which allow many volunteers to be studied at the same time, have proven useful in the evaluation of drugs that afford acute symptom relief. However, such challenges incompletely model naturally occurring disease, in which the repeated daily exposure to allergen modifies the mucosal inflammatory cell profile and in particular promotes the epithelial accumulation of effector cells. This alters the response to allergen exposure. To model this, repeated low-dose daily allergen exposure has been used to generate these mucosal changes artificially, and early studies suggest that this may be a more valid model for the evaluation of anti-inflammatory therapy. However, little has been published with this model. Different disease groups are associated with their own specific issues in clinical trials. The pediatric population, in which allergic rhinitis is common, has different requirements for education, quality of life evaluation, and adverse-event monitoring; nasal polyposis, because of the nature of the disease, requires additional means of assessment, such as nasal endoscopy and imaging (eg, computerized tomography scanning), as well as attention to additional outcome measures (eg, the measurement of sense of smell). Within clinical trial design, there are important questions to be considered in relationship to the therapeutic intervention. Should this be given topically or systemically? What are the appropriate timing and frequency of medication? Does the disease itself modify the treatment efficacy, and does combination therapy afford better clinical outcome than single-modality therapy? These issues are discussed, and the influences of current therapies on objective outcome measures in allergic rhinitis are reviewed. PMID- 15746884 TI - Endocrine disrupters as disrupters of brain function: a neurosteroid viewpoint. AB - The mechanisms of neurosteroid synthesis in the rat hippocampus were investigated. Metabolism assay demonstrated the pathway of "cholesterol alpha pregnenolone --> dehydroepiandrosterone --> androstenedione --> testosterone --> estradiol." Upon exposure of pups to bisphenol A (BPA) from the embryonic stage until 3 week-old stage, a significant facilitation of the synthesis of estradiol was observed in the hippocampus. The localization of cytochrome P450s (P450scc, P45017alpha, and P450arom) as well as estrogen receptor alpha (ER(alpha)) was observed in pyramidal and granule neurons, using immunohistochemical staining. Furthermore, the synaptic localization of P45017alpha, P450arom and ER(alpha) was demonstrated with immuno-electron microscopic analysis. The acute action of estradiol and endocrine disrupters were then analyzed with an electrophysiological measurement of hippocampal pyramidal neurons. A 30 min preperfusion of diethylstylbesterol (DES) enhanced the induction of long-term potentiation (LTP) by almost an identical magnitude to that obtained by estradiol perfusion. On the other hand, although the application of BPA alone did not affect LTP-induction, the co-perfusion of BPA with estradiol completely suppressed the enhancement effect of LTP by estradiol. The current investigations demonstrate in the hippocampus (1) that locally synthesized estrogen rapidly enhances the synaptic plasticity of neurons, and (2) that BPA and DES modulate the synaptic plasticity as well as the synthesis of estradiol. The probable targets of BPA and DES are ER(alpha) and steroidogenic proteins. PMID- 15746885 TI - Development of a GC-MS system with an array detector system for endocrine disruptor analysis. AB - A mass spectrograph using a position sensitive detector was designed and constructed. It has an image contractor lens system and a zoom lens system. The position sensitive detector consists of a micro channel plate (MCP) and a charge coupled device (CCD). The performance of this new instrument was tested with methylstearate as a sample. PMID- 15746886 TI - Technical aspects of gel-based proteomics designed for elucidating an aryl hydrocarbon receptor complex. AB - The identification of proteins by mass spectrometry has revolutionalized the basic method of identifying proteins constituting an intracellular unit or network for certain biological functions. The gel-based strategy following immunoprecipitation was applied to elucidating proteins associated with the aryl hydrocarbon receptor (AhR). Two hundred femtomoles of AhR was recovered from approximately 2 x 10(7) HepG2 cells by immunoprecipitation and was sufficient for identification by peptide mass fingerprinting. Possible candidates for the AhR associated proteins were also identified. Improvements of the current strategy to increase the overall sensitivity tenfold are required to clarify the AhR complex in full detail. For example, a combination of trypsin and Achromobacter protease I for in-gel digestion allows the number of missed cleavage sites to be set at zero for database searching, thereby reducing random matches and facilitating identification. There is also room for improvement in each step of sample preparation prior to mass spectrometry. PMID- 15746887 TI - Endocrine-disrupting activity of chemicals in diesel exhaust and diesel exhaust particles. AB - Diesel exhaust (DE) is known as the main cause of air pollution. DE is a complex mixture of particulate and vapor-phase compounds. The soluble organic fraction of the particulate materials in DE contains thousands of compounds including a variety of polycyclic aromatic hydrocarbons and heavy metals. To clarify the endocrine-disrupting activities of DE, we have reviewed the reports about the effects of DE on the reproductive and brain-nervous systems, and the endocrine disrupting action of diesel exhaust particles (DEP). In utero exposure to low levels (0.1 mg DEP/m3) of DE from day 2 postcoitum (p.c.) until day 13 p.c. reduced the expression level of Ad4BP/SF-1 mRNA and thereby might affect the development of gonads. Low levels of DE also reduced the expression of several genes known to play key roles in gonadal development, including an enzyme necessary for testosterone synthesis. Mature male rats exposed to DE during the fetal period showed an irreversible decrease in daily sperm production due to an insufficient number of Sertoli cells. DE exposure during the fetal period influenced the brain tissue in newborn mice. In the 3 mg DEP/m3 exposure group at 10 weeks of age, a significant reduction in performance was observed in the passive avoidance learning test in both male and female mice. In addition, the fetal exposure of mice to DE affected the emotional behaviors associated with the serotonergic and dopaminergic systems in the mouse brain. In toluidine blue stained specimens from the DE-exposed group, edema around the vessels where fluorescent granular perithelial (FGP) cells exist and degenerated granules within the FGP cytoplasm were observed; similar findings were obtained by electron microscopic examination. DEP contain many substances that stimulate Ah receptors, such as the polycyclic aromatic hydrocarbon containing benzo[a]pyrene. DEP also contain substances with estrogenic, antiestrogenic and antiandrogenic activities. The neutral substance fraction of DEP has the causal substance that reduces estrogen receptor mRNA expression. Evaluating the influence of these chemicals present in the environment on human health is an important task. PMID- 15746888 TI - Effects of dioxin on gene expression in female reproductive system in the rat. AB - We analyzed the effects of a low dose of dioxin on gene expression in reproductive tissues or cells by microrepresentation differential analysis (micro RDA) and DNA microarray analysis, and identified more than one hundred dioxin sensitive genes in cultured rat granulosa cells, rat placentas and rat ovaries. It is well known that dioxin exerts effects mainly through Ah receptors which recognize the xenobiotic responsive element (XRE) in the promoter region of target genes. The present data, however, indicate that a low dose of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) induces or suppresses the expression of various genes, which have not been known as dioxin-sensitive. These data suggest that a low dose of dioxin has profound effects on the reproductive system. PMID- 15746889 TI - Effect of insulin-like growth factor-1 and 2,3,7,8-tetrachlorodibenzo-p-dioxin on the expression of luteinizing hormone receptors in cultured granulosa cells. AB - In this study, we attempted to assess the effects of insulin-like growth factor-1 (IGF-1) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on follicle-stimulating hormone (FSH)-induced luteinizing hormone (LH) receptor expression in rat granulosa cells to understand the effects of these factors on normal reproductive function. Treatment with FSH, as expected, produced a substantial increase in LH receptor mRNA expression level, and cotreatment with an increasing concentration of IGF-1 resulted in a dose-dependent increase in FSH-induced LH receptor mRNA expression level. On the other hand, the cotreatment with FSH and TCDD (10 pM) resulted in a significant decrease in LH receptor mRNA expression level after 24 h. The decay curves for the LH receptor mRNA transcript showed a significant increase in half-life after the addition of IGF-1 and a significant decrease after the addition of TCDD. These data suggest a possible role for changes in LH receptor mRNA stability in the IGF-1- and TCDD-induced regulation of the LH receptor in rat granulosa cells. The rates of LH receptor mRNA gene transcription, assessed by nuclear run-on transcription assay, did not increase after the addition of IGF-1, but decreased after the addition of TCDD. The data for IGF-1 indicate that the interface between circulating hormones and paracrine/autocrine systems could provide an important mechanism for amplifying the effects of gonadotropin hormones at the local level. In addition, the endocrine-disrupting effects of TCDD are, at least in part, caused by the direct action on the LH receptor expression on granulosa cells. PMID- 15746890 TI - Sexual plasticity in fish: a possible target of endocrine disruptor action. AB - Various genetic and molecular approaches have been used to investigate the mechanisms of sex determination, gonadal sex differentiation, and sex change in fish. We identified, for the first time in nonmammalian vertebrates, DMY, as the sex-determining gene of medaka. In tilapia, endogenous estrogens act as the natural inducers of ovarian differentiation, while DMRT1 may be important for testicular differentiation. In the protogynous wrasse, a rapid decline in serum estradiol-17beta levels may be an initial trigger of the female-to-male sex change. Both sex steroids and endocrine disrupters do not seem to act at the level of the sex-determining gene, but during gonadal sex differentiation. The Japanese flounder exhibits temperature-dependent sex determination. Some of the estrogenic endocrine disrupters induce feminization of the flounder larvae reared at the masculinizing temperature. The actions of these sex steroids and endocrine disrupters may be mediated by the actions of somatic cells within gonads. Thus, sexual plasticity of gonads during sex differentiation may be implicated through the somatic cells within gonads. Cloning and sequencing of a number of genes that are considered to be associated with gonadal sex differentiation have been performed and some are still in progress. These molecular probes provide useful tools for understanding not only the molecular mechanisms of sex determination and gonadal sex differentiation but also provide important basic information for studying the effects of endocrine-disrupting chemicals during these periods. PMID- 15746891 TI - An epidemiological study of the incidence of abnormal pregnancy in areas heavily contaminated with methylmercury. AB - To determine the relationship between abnormal pregnancy in humans and methylmercury contamination, a retrospective study was conducted on women in two heavily contaminated areas, Modo (area M) and Akasaki (area A). Abnormal pregnancy was defined as fetal death, i.e., stillbirth and spontaneous abortion. In area M, prior to 1956, the incidence of abnormal pregnancy among the respondents was 7.0%. The incidence increased to 18.1% in the period between 1956 and 1968, when the pollution become serious. In area A, the incidence increased from 5.4% before 1956 to 14.2% between 1956 and 1968. In each area, the difference in the incidence between two periods, i.e., before and during the severe contamination, was statistically significant (area M, p<0.01; area A, p<0.001). Women living in Ikitsuki Island (area I) were selected as the control group. The incidence of the abnormal pregnancy in the three areas was compared by the birth years of mothers. Among women born between 1931 and 1940, the incidences were 26%, 15.1% and 13.5% in areas M, A and I, respectively. These women reached their reproductive age during the period of severe contamination in the two areas. The differences in the incidence between areas M and A and between areas M and I were statistically significant (areas M and A, p<0.05; areas M and I, p<0.01). These suggest that there is a relationship between methylmercury contamination and the increase in abnormal pregnancy in humans. PMID- 15746892 TI - Development of vitellogenin assay for endocrine disrupters using medaka (Oryzias latipes). AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the egg yolk precursor vitellogenin (VTG) in the liver of medaka (Oryzias latipes) and was employed to establish an in vivo testing system for estrogen and estrogenic compounds using liver homogenates. Results of 3-month-old fish exposed to three reference chemicals (ethynylestradiol, methyltestosterone and flutamide) recommended by Organisation for Economic Co-operation and Development (OECD) for the validation showed the induction and decrease of vitellogenic responses, making the assay using the liver VTG of medaka a possible screening method for not only estrogens but also androgens. In addition, 21-day exposure of male fish to 4-tert-octylphenol and 4-nonylphenol produced concentration-dependent inductions of liver vitellogenin, with the lowest observed effect concentrations of 64.1 microg/L and 22.5 microg/L, respectively, while no significant VTG responses were observed in male and female fish exposed to tributyltin chloride and dibutyl phthalate. This study demonstrates that the VTG assay using liver homogenates from small fish such as medaka can be used as a screening method for environmental estrogens. This is because the sensitivity of the VTG response in medaka may be almost the same as that of other fish using blood samples. PMID- 15746893 TI - Distribution of tributyltin, dibutyltin and monobutyltin in the liver, brain and fat of rats: two-generation toxicity study of tributyltin chloride. AB - The distribution of tributyltin (TBT) and its metabolites, dibutyltin (DBT) and monobutyltin (MBT), was examined in the liver, brain and fat tissues in a two generation reproductive toxicity study of tributyltin chloride (TBTCl) in rats using dietary supplementation at concentrations of 5, 25 and 125 ppm. In the liver, irrespective of TBTCl dietary concentration, gender or generation, the highest concentration of metabolite was consistently MBT, followed by DBT, and then TBT. In contrast, TBT was consistently present at the highest concentration in the brain, nearly always followed by DBT and MBT. In fat tissues, the concentrations of the three butyltin compounds showed similar relationships to those observed in the brain, although the concentrations were much lower. In the liver, the concentration of TBT was higher in females, and those of DBT and MBT were higher in males. Factorial ANOVA also suggested the effect of gender on the concentrations of the three butyltin compounds in the liver. The results of this study suggest tissue-dependent distribution of TBT, DBT and MBT and gender dependent distribution of the three metabolites in the liver of rats. PMID- 15746894 TI - Effects of triclosan on various aquatic organisms. AB - Triclosan (2,4,4'-trichloro-2'-hydroxydiphenyl ether) is widely used as an antibacterial agent in various industrial products, such as textile goods, soap, shampoo, liquid toothpaste and cosmetics, and often detected in wastewater effluent. However, there is a paucity of data on the toxicity of triclosan and its effects on aquatic organisms. In this study, the acute toxicity of triclosan to the Microtox bacterium (Vibrio fischeri), a microalga (Selenastrum capricornutum), a crustacean (Ceriodaphnia dubia) and fish (Danio rerio and Oryzias latipes) was examined. As a result, the MicrotoxR bacterium, crustacean and fish had similar sensitivities towards triclosan toxicity (i.e., IC25 from 0.07 to 0.29 mg/L triclosan). In contrast, the microalga was about 30-80-fold (IC25 = 0.0034 mg/L triclosan) more sensitive to triclosan toxicity than the bacterium and fish. Therefore, triclosan is quite highly toxic to aquatic animals, and is particularly highly toxic to the green alga used as a test organism in this study. This result indicates that triclosan exerts a marked influence on algae, which are important organisms being the first-step producers in the ecosystem; therefore, the possible destruction of the balance of the ecosystem is expected if triclosan is discharged into the environment at high levels. PMID- 15746895 TI - Near-UV radiation promotes growth of Chlorella. AB - The wavelength of natural sunlight reaching the Earth's surface was detected to be above 300 nm using a multichannel spectrodetector and the ratio of UV to photosynthetically active radiation (PAR) of sunlight in Japan (137 degrees E/35 degrees 11' N, altitude; 50 m) in early summer was estimated to be 0.07:1 (i.e., 7%). On the basis of this UV/PAR ratio, Chlorella ellipsoidea (IAM-27) cells were cultured in flasks under various conditions of UV irradiation in growth chambers. The growth (cell division) of these cells without near-UV radiation was inferior to that with near-UV radiation. Growth at a UV/PAR ratio of 7% (natural conditions), determined tentatively using our detector in the present study, was maximal similar to those at 14% and 28%; whereas that at 0.7% was somewhat less and that at 70% was considerably less. Growth was linked with the activities of stress enzymes. NAD(P)H-dependent oxidase [NAD(P)-DH] and xanthine oxidase (XOD), extracted from Chlorella exposed to near-UV radiation demonstrated lower activities than those from Chlorella not exposed to near-UV radiation. On the other hand, superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APOD) extracted from Chlorella exposed to near-UV radiation have higher activities than those from unexposed Chlorella. Near-UV radiation clearly acted as an important factor for growth (cell division), at least at UV/PAR ratios of up to 0.28:1. PMID- 15746896 TI - Regulation of ethylene-forming system in Chlorella by near-UV radiation. AB - The growth (cell division) of Chlorella cells cultured under photosynthetically active radiation (PAR) without near-UV radiation was inferior to that under PAR with near-UV radiation. To elucidate this phenomenon, the relationship between near-UV radiation and ethylene production in Chlorella cells was examined. The suppression of ethylene production by UV radiation suggests that this phenomenon is associated with the production of ethylene. Chlorella (a eukaryotic protista) was found to produce ethylene from methionine via S-adenosyl-methionine (SAM) and 1-aminocyclo-propane-1-carboxylate (ACC) as in higher plants, and the activity of ACC oxidase, a limiting factor of ethylene production, appears to be associated with the growth of Chlorella. A possible mechanism for the action of near-UV radiation on ethylene biosynthesis and growth suppression by ethylene is discussed. PMID- 15746897 TI - Altered metabolism of dopamine in the midbrain of mice treated with tributyltin chloride via subacute oral exposure. AB - Tributyltin (TBT) compounds have been detected in fish and shellfish. One of the targets of TBT compounds is the central nervous system. Alterations in the levels of neurotransmitters and their metabolites, and ratios of the levels of neurotransmitters to those of their metabolites have been used as indexes of neurotoxicity. We evaluated the neurotoxicity of TBT compounds in mice following subacute oral exposure by determining the levels of neurotransmitters and their metabolites in discrete brain regions. Male BALB/c mice were exposed to 0, 1, 5, 25, or 125 ppm TBT chloride in their feed for one month. Following the treatment period, their brains were removed and dissected into the cerebrum, cerebellum, medulla oblongata, midbrain, corpus striatum and hypothalamus. The levels of norepinephrine, dopamine (DA), dihydoxyphenylacetic acid, homovanillic acid (HVA), serotonin, and 5-hydroxyindolacetic acid were determined in different brain regions by high-performance liquid chromatography (HPLC). The mean body weight of mice treated with 125 ppm TBT was significantly lower than that of the control from day 5 to day 16 during the treatment period. The HVA/DA ratio in the midbrain of the 125 ppm-treated group was significantly higher than those of other treatment groups, and tended to be higher than that of the control. TBT may affect DA metabolism in the brain, especially in the midbrain. PMID- 15746898 TI - Quantification of vitellogenin in several developmental stages of medaka (Oryzias latipes) S-rR strain. AB - Concentrations of vitellogenin (VTG) in different developmental stages: embryo (on the day of fertilization); yolk-sac larva (on the day of hatching); 2, 4, 6 and 8 weeks posthatching; were determined in medaka (Oryzias latipes) S-rR strain. Both sexes were bred separately from 1 week after hatching and fed only Artemia nauplii to avoid contamination by xenoestrogen from females and feed. Whole bodies of ten test fish/group were homogenized individually and the supernatants were quantified. Quantification of VTG was performed by an enzyme linked immunosorbent assay (ELISA). In most males, VTG levels were less than 1 microg/g body weight and no fluctuation was observed throughout the developmental stages. In 2-week-old females, five had no detectable VTG, and the others had 5.18+/-2.37 microg/g of VTG. The 4-week-old females had 16.3+/-12.0 microg/g VTG, and the concentrations increased with maturity to 5.54+/-3.09 mg/g in 6-week-old and 8.99+/-3.10 mg/g in 8-week-old specimens. These results show that the concentrations of VTG in males are routinely close to the detection limit independent of the developmental stages in an environment with low contamination by xenoestrogen derived from artificial feed and females. Females have detectable VTG levels even in the juvenile phase, and the level increases markedly with maturation. PMID- 15746899 TI - cDNA cloning, sequence analysis and expression of 3-methylcholanthrene-inducible cytochrome P450 1B1 in carp (Cyprinus carpio). AB - Cytochrome P450 (CYP) genes, which make up a large gene superfamily, are known to play an important role in drug metabolism. The levels of expression of CYP genes in the tissue of fish inhabiting polluted areas have been used extensively in biomonitoring studies as indicators of dioxin pollution. Complementary DNA of cytochrome CYP1B1 was isolated from carp (Cyprinus carpio) liver 24 h after the injection of 3-methylcholanthrene (3-MC). The full length cDNA obtained contained a 5' noncoding region of 178 bp, an open reading frame of 1593 bp coding for 530 amino acids and a stop codon, and a 3' noncoding region of 1599 bp. The predicted molecular weight of the encoded protein was approximately 60.2 kDa. The deduced amino acid sequence exhibited an identity of 60.6% with reported CYP1B sequences of plaice CYP1B, and of 52.4, 51.4, and 50.3% with human, rat, and mouse CYP1B1s, respectively. It exhibited similarities of 48.4 and 47.3% with scup CYP1C2 and 1C1 sequences. The percent identities with CYP1A sequences showed lower values in the range from 35.3 to 39.5% with mammals and teleosts. The phylogenetic tree of the CYP1 family members constructed by the protein maximum likelihood method indicates that the carp CYP1B1 and plaice CYP1B share a common ancestry with the mammalian CYP1B1s. Carp treated with 3-MC showed expression of CYP1B1 in liver, intestine and gills with distinct induction except for the gills that showed marked constitutive expression. The presence of two successive signals in treated gills at low stringency hybridization may suggest the existence of another CYP1B member that is expressed in the gills of carp. PMID- 15746900 TI - Seasonal changes in sex ratio, maturation, and size composition of fresh water snail, Sinotaia quadrata histrica, in Lake Kasumigaura. AB - Sinotaia quadrata histrica is a fresh water viviparous snail distributing from the Kanto region to Kyushu Island, Japan. About 7000 snails were collected in Lake Kasumigaura (L. Nishiura and L. Kitaura) in 2001 and 2002, and the sex ratio, maturity in terms of the gonad-somatic index (GSI) and operculum diameter were determined. The total female proportion was 55.2% in 2001, 53.0% in 2002 in L. Nishiura, and that of L. Kitaura was 60.4% in 2002. Comparing the season, the female proportion was the highest during early summer in both 2001 (59.6%, July in L. Nishiura) and 2002 (61.6%, June in L. Nishiura, 65.8%, July in L. Kitaura). The GSI of females in L. Nishiura significantly increased from April to May and significantly decreased from June to August. The GSI of males was higher in spring, but significantly lower from June to August. The mean female operculum diameter was consistently larger than that of males for each month and year, and a particularly significant difference was found between females and males from April to August 2001, and from April to September 2002. The number of resting zones on the operculum correlates with the operculum diameter and the female proportion was larger in the snails, which have a high number of resting zones, suggesting a sex-dependent difference in age composition. This study estimated that the sex ratio and seasonal maturation of S. quadrata histrica and the sex dependent difference in age composition might contribute to the population structure in L. Kasumigaura. PMID- 15746901 TI - Effects of 17beta-estradiol on survival, growth, sexual development and molting cycles of the marine crustacean mysid shrimp, Americamysis bahia. AB - A 14-day partial life-cycle test was performed to assess the effects of 17beta estradiol (E2) on the survival, growth, sexual development and molting cycles of a marine crustacean mysid shrimp (Americamysis bahia). Seven-day-old mysids were exposed to the nominal E2 concentrations of 31.3, 62.5, 125, 250 and 500 microg/l for 14 days. The total length and the body weight of mysids significantly decreased relative to the controls when exposed to 62.5, 250 and 500 microg/l E2 for 14 days. Moreover, the carapace length significantly decreased in the 500 microg/l E2 treatment groups. No significant differences were observed in sex ratio with the appearance of secondary sex characteristics in the all treatment groups including the control and solvent control groups. However, the percentage of females with eggs in the oviduct or brood sac decreased significantly in mysids treated with E2 at 62.5, 125, 250 and 500 microg/l. The cumulative total number of molting cycles when exposed to E2 for 14 days significantly decreased in the treatment groups at 500 microg/l relative to the controls. These results suggest that concentrations of E2 over 62.5 microg/l may cause growth suppression in mysid shrimp, and that the disruption of molting cycles may result in alterations in growth due to a sublethal response to toxicant exposure. PMID- 15746902 TI - Multiple reporter gene assays for the assessment and estimation of chemical toxicity. AB - To detect chemical toxicity, we are making new bioassay systems that use promoters selected from yeast DNA microarray experiments. We performed multiple reporter gene assays using the promoters of these genes; the promoter regions were inserted upstream of green fluorescence protein (GFP). In this report, six genes (HSP26, MET17, YLL057C, FIT2, CUP1 and OYE3) were selected and assays were carried out for 55 chemicals. The promoters of these genes showed different responses to chemicals within 4 h. This result indicates that this technique enables us to predict the toxicity of chemicals in the environment and to understand toxicities of newly synthesized chemicals. PMID- 15746903 TI - Differences in responses of plant hairy roots to chemical toxicity compared between primary and ramified roots. AB - As a bioassay system for the assessment of the chronic toxicity of chemicals, successive culturing of pak-bung hairy roots was conducted with serial passaging of the primary and lateral root tips. Based on the elongation rates of these root tips, the dose-response profiles were examined while exposing the roots to 2,4 dichlorophenoxyacetic acid (2,4-D) and 1-methyl-3-nitro-1-nitrosoguanidine (MNNG) at concentrations of 0.01-3.0 micromol/dm3, and the values of the median effective concentration, EC50, were determined under the conditions examined. Irrespective of the test chemicals, the EC50 values of both the primary and lateral roots gradually decreased with increasing exposure period ranging from 7 to 35 days. A marked difference in the EC50 values was observed between the primary and lateral roots exposed to 2,4-D; the EC50 value of the lateral roots following exposure for 35 days was 0.036 micromol/dm3, which was one-eighth that of the primary roots for the same exposure period. Moreover, it was found that this phenotypic difference in the root responses was consistently reflected in the distinct patterns of DNA fingerprints of the primary and lateral roots analyzed by the random amplified polymorphic DNA method. PMID- 15746904 TI - Development of a genotoxicity detection system using a biosensor. AB - The umu-lux test is a genotoxicity test using the two genetically modified S. typhmurium TA1535 strains (TL210 and TL210ctl) transformed with the luxCDABE (luciferase gene and fatty acid reductase genes) of Vibrio fischeri as a reporter gene. The TL210 strain detects genotoxicants and the TL210ctl strain detects cytotoxicants. In order to develop a highly sensitive, simple and rapid genotoxicity detection system, we constructed a biosensor using these immobilized strains. The biosensor consists of two immobilized microbial membranes, a sample vessel and photodetectors, and the genotoxicity detection system consists of the biosensor, an isothermal box, a photodetector and an air pump. The total measurement time for genotoxicants using this detection system is about 4 h. When 2% (v/v) DMSO was used as a control, the TL210 strain was not emitting light while the TL210ctl strain was. When 0.3 mg/l 4NQO was used as a genotoxicant, TL210 strain and TL210ctl strain were both emitting light. When HgCl2 was used as a cytotoxicant, neither the TL210 strain nor the TL210ctl strain were emitting light. Therefore, the false negative prevention function of a biosensor using the TL210ctl strain has been checked. These results show that our proposed system can correctly detect genotoxicants. PMID- 15746905 TI - Separating science from stereotype. PMID- 15746907 TI - Inhibitory synapses turn exciting. PMID- 15746908 TI - Stem cells in the injured spinal cord: reducing the pain and increasing the gain. PMID- 15746909 TI - Controlling stress: how the brain protects itself from depression. PMID- 15746910 TI - Myelin repair: developmental myelination redux? PMID- 15746911 TI - Zooming in on cortical maps. PMID- 15746912 TI - Disentangling simple from complex cells. PMID- 15746913 TI - Constant light desynchronizes mammalian clock neurons. AB - Circadian organization can be disrupted by constant light, resulting in behavioral arrhythmicity or 'splitting' of rhythms of activity and rest. By imaging molecular rhythms of individual clock neurons in explanted mouse clock nuclei, we now find that constant light desynchronizes clock neurons but does not compromise their ability to generate circadian rhythms. Cellular synchrony within clock nuclei is disrupted during arrhythmicity, whereas neurons in the left and right clock nuclei cycle in antiphase during 'splitting.' PMID- 15746918 TI - Bad medicine. PMID- 15746914 TI - Sonic hedgehog guides commissural axons along the longitudinal axis of the spinal cord. AB - Dorsal commissural axons in the developing spinal cord cross the floor plate, then turn rostrally and grow along the longitudinal axis, close to the floor plate. We used a subtractive hybridization approach to identify guidance cues responsible for the rostral turn in chicken embryos. One of the candidates was the morphogen Sonic hedgehog (Shh). Silencing of the gene SHH (which encodes Shh) by in ovo RNAi during commissural axon navigation demonstrated a repulsive role in post-commissural axon guidance. This effect of Shh was not mediated by Patched (Ptc) and Smoothened (Smo), the receptors that mediate effects of Shh in morphogenesis and commissural axon growth toward the floor plate. Rather, functional in vivo studies showed that the repulsive effect of Shh on postcommissural axons was mediated by Hedgehog interacting protein (Hip). PMID- 15746915 TI - Inhibitory synapses in the developing auditory system are glutamatergic. AB - Activity-dependent synapse refinement is crucial for the formation of precise excitatory and inhibitory neuronal circuits. Whereas the mechanisms that guide refinement of excitatory circuits are becoming increasingly clear, the mechanisms guiding inhibitory circuits have remained obscure. In the lateral superior olive (LSO), a nucleus in the mammalian sound localization system that receives inhibitory input from the medial nucleus of the trapezoid body (MNTB), specific elimination and strengthening of synapses that are both GABAergic and glycinergic (GABA/glycinergic synapses) is essential for the formation of a precise tonotopic map. We provide evidence that immature GABA/glycinergic synapses in the rat LSO also release the excitatory neurotransmitter glutamate, which activates postsynaptic NMDA receptors (NMDARs). Immunohistochemical studies demonstrate synaptic colocalization of the vesicular glutamate transporter 3 with the vesicular GABA transporter, indicating that GABA, glycine and glutamate are released from single MNTB terminals. Glutamatergic transmission at MNTB-LSO synapses is most prominent during the period of synapse elimination. Synapse specific activation of NMDARs by glutamate release at GABAergic and glycinergic synapses could be important in activity-dependent refinement of inhibitory circuits. PMID- 15746920 TI - Tsunami aid spending draws fire over public health. PMID- 15746921 TI - Strict NIH ethics rules provoke mixed response from scientists. PMID- 15746922 TI - Activists angry at fallout from AIDS drug trial allegations. PMID- 15746923 TI - Action plan peps up Europe's mental health. PMID- 15746926 TI - Antiaging drug trials compel creative testing methods. PMID- 15746927 TI - HIV drug program likely to miss 2005 target. PMID- 15746928 TI - Profile: Mina Bissell. PMID- 15746929 TI - A crucial test. PMID- 15746930 TI - Reports on nevirapine threaten public health. PMID- 15746932 TI - Sound from silence. PMID- 15746933 TI - Sensing the danger in RNA. PMID- 15746934 TI - Multiple sclerosis: trapped in deadly glue. PMID- 15746935 TI - Gut-level decisions in peace and war. PMID- 15746936 TI - The need for natural killer T cells. PMID- 15746937 TI - Part-time cop nabs deviant DNA. PMID- 15746940 TI - Histone deacetylase inhibitors differentially stabilize acetylated p53 and induce cell cycle arrest or apoptosis in prostate cancer cells. AB - In LNCaP prostate cancer cells CG-1521, a new inhibitor of histone deacetylases, alters the acetylation of p53 in a site-specific manner. While p53 is constitutively acetylated at Lys320 in LNCaP cells, treatment with CG-1521, stabilizes the acetylation of p53 at Lys373, elevating p21 (and inducing cell cycle arrest). Treatment with CG-1521 also promotes Bax translocation to the mitochondria and cleavage, and apoptosis. TSA stabilizes the acetylation of p53 at Lys382, elevating p21 levels and inducing cell cycle arrest, but does not induce Bax translocation or apoptosis. In LNCaP cells CG-1521, but not TSA, promotes the rapid degradation of HDAC2. These data suggest that the acetylation of p53 at Lys373 is required for the p53-mediated induction of cell cycle arrest and apoptosis, while acetylation of p53 at Lys382 induces only cell cycle arrest. In p53(-/-) PC3 cells both compounds induce p21 and cell cycle arrest, but not Bax translocation or apoptosis, suggesting that both compounds can also induce p21 through a p53-independent mechanism. PMID- 15746941 TI - Triterpenoid CDDO-Im downregulates PML/RARalpha expression in acute promyelocytic leukemia cells. AB - The triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) induces differentiation and apoptosis of diverse human tumor cells. In the present study, we examined the effects of the CDDO imidazolide imide (CDDO-Im) on the NB4 acute promyelocytic leukemia (APL) cell line and primary APL cells. The results show that CDDO-Im selectively downregulates expression of the PML/retinoic receptor alpha fusion protein by a caspase-dependent mechanism and sensitizes APL cells to the differentiating effects of all-trans retinoic acid (ATRA). CDDO-Im treatment of APL cells was also associated with disruption of redox balance and activation of the extrinsic apoptotic pathway. In concert with these results, CDDO-Im sensitizes APL cells to arsenic trioxide (ATO)-induced apoptosis. Our findings indicate that CDDO-Im may be effective in the treatment of APL by: (i) downregulation of PML/RARalpha; (ii) enhancement of ATRA-induced differentiation; and (iii) sensitization of ATO-induced APL cell death. PMID- 15746942 TI - PGE(2) in the regulation of programmed erythrocyte death. AB - Hyperosmotic shock, energy depletion, or removal of extracellular Cl(-) activates Ca(2+)-permeable cation channels in erythrocyte membranes. Subsequent Ca(2+) entry induces erythrocyte shrinkage and exposure of phosphatidylserine (PS) at the erythrocyte surface. PS-exposing cells are engulfed by macrophages. The present study explored the signalling involved. Hyperosmotic shock and Cl(-) removal triggered the release of prostaglandin E(2) (PGE(2)). In whole-cell recording, activation of the cation channels by Cl(-) removal was abolished by the cyclooxygenase inhibitor diclophenac. In FACS analysis, phospholipase-A(2) inhibitors quinacrine and palmitoyltrifluoromethyl-ketone, and cyclooxygenase inhibitors acetylsalicylic acid and diclophenac, blunted the increase of PS exposure following Cl(-) removal. PGE(2) (but not thromboxane) induced cation channel activation, increase in cytosolic Ca(2+) concentration, cell shrinkage, PS exposure, calpain activation, and ankyrin-R degradation. The latter was attenuated by calpain inhibitors-I/II, while PGE(2)-induced PS exposure was not. In conclusion, hyperosmotic shock or Cl(-) removal stimulates erythrocyte PS exposure through PGE(2) formation and subsequent activation of Ca(2+)-permeable cation channels. PMID- 15746943 TI - Development of an inducible suicide gene system based on human caspase 8. AB - Suicide gene-therapy strategies are promising approaches in treating various diseases such as cancers, atherosclerosis, and graft-versus-host-disease. Here, we describe the development of a new effector gene based on inducing functional caspase 8, the initiator caspase in the death-receptor pathway. We constructed vectors encoding a constitutively active form of human caspase 8 (CC8), and demonstrated the efficient killing of a variety of cell types in transfection and lentivirus-transduction assays. We then analyzed the ability to control the apoptotic activity of a caspase 8-derived construct through the ARIADtrade mark homodimerization system (FKC8), a system shown to be extremely effective in several cellular models upon retroviral and lentiviral gene transfer. Similarly, two transcription-regulation systems, muristerone-regulated and Tet-On, were tested to control the expression of CC8. The homodimerization-regulated system FKC8 was shown to be the most efficient system with low background activity in noninduced conditions. In the presence of a dimerizer, it was as active as the activated Tet-On system. From our data, we conclude that the dimerizer-dependent human caspase 8 represents a highly inducible and very powerful system to eradicate transduced cell populations. In addition to its application in experimental gene therapy, this variant may be highly useful for mechanistic research related to apoptosis. PMID- 15746944 TI - Transcriptional targeting of RGD fiber-mutant adenovirus vectors can improve the safety of suicide gene therapy for murine melanoma. AB - Since RGD fiber-mutant adenovirus vector (AdRGD), which contains an alphav integrin tropism, is highly efficient in gene transduction to melanoma, the AdRGD mediated herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) system is an attractive approach for melanoma treatment. However, the intratumoral injection of AdRGD causes limited transgene expression in healthy normal tissue, due to unwanted vector spread. Herein, we describe our attempt to overcome this limitation related to the safety of HSVtk/GCV treatment by using AdRGD carrying either melanoma-specific tyrosinase (Tyr) promoter or tumor-specific telomerase reverse transcriptase (TERT) promoter instead of universal cytomegalovirus promoter. Our in vitro study revealed that Tyr promoter-regulated AdRGD exhibited high transgene expression specificity for melanoma cells, and that TERT promoter regulated AdRGD could induce efficient gene expression in tumor cells, but was relatively quiescent in normal cells. Anti-B16BL6 melanoma effects in mice injected intratumorally with AdRGD-Tyr/HSVtk or AdRGD-TERT/HSVtk, after which GCV was injected intraperitoneally for 10 days, were comparable to those in mice injected with AdRGD-CMV/HSVtk at 10 times less vector dosage. On the other hand, AdRGD-Tyr/HSVtk and AdRGD-TERT/HSVtk did not induce severe adverse effects even when they were intravenously injected into mice at 10(9) plaque-forming units (PFU), whereas mice injected with AdRGD-CMV/HSVtk at 10(8) PFU exhibited body weight reduction and serum level increase of biochemical enzymes for hepatotoxicity. These results indicate that AdRGD combined with transcriptional regulation using Tyr or TERT promoter is a potentially useful and safe vector system for suicide gene therapy for melanoma. PMID- 15746945 TI - Oncolytic activities of approved mumps and measles vaccines for therapy of ovarian cancer. AB - Oncolytic viruses are promising cytoreductive agents for cancer treatment but extensive human testing will be required before they are made commercially available. Here, we investigated the oncolytic potential of two commercially available live attenuated vaccines, Moraten measles and Jeryl-Lynn mumps, in a murine model of intraperitoneal human ovarian cancer and compared their efficacies against a recombinant oncolytic measles virus (MV-CEA) that is being tested in a phase I clinical trial. The common feature of these viruses is that they express hemagglutinin and fusion therapeutic proteins that can induce extensive fusion of the infected cell with its neighbors, resulting in death of the cell monolayer. In vitro, the three viruses caused intercellular fusion in human ovarian cancer cells but with marked differences in fusion kinetics. MV-CEA was the fastest followed by Jeryl-Lynn mumps virus while Moraten measles virus was the slowest, although all viruses eventually caused comparable cell death 6 days postinfection. Tumor-bearing mice treated with 10(6) or 10(7) pfu (one thousand times the vaccine dose) of each of the three viruses responded favorably to therapy with significant prolongations in survival. All three viruses demonstrated equivalent antitumor potency. Commercially available Moraten measles and Jeryl-Lynn mumps vaccines warrant further investigation as potential anticancer agents. PMID- 15746946 TI - A virus-directed enzyme prodrug therapy (VDEPT) strategy for lung cancer using a CYP2B6/NADPH-cytochrome P450 reductase fusion protein. AB - Virus-directed enzyme prodrug therapy (VDEPT) is an emerging strategy against cancer. Our approach is a P450-based VDEPT that consists of using cyclophosphamide (CPA) as a prodrug and a Cytochrome P450 2B6/NADPH cytochrome P450 reductase fusion protein (CYP2B6/RED) as a prodrug-activating enzyme. Due to the heterogenous expression of proteins in tumor cells, basal reductase activity may not be sufficient to supply CYP2B6 with electrons, the fusion protein should enable the expression of both proteins at high levels in tumor cells. CYP/RED fusion proteins have never been previously expressed in mammalian cells, to enable expression the fusion protein was cloned into an adenoviral vector and subsequently several pulmonary tumor cell lines were infected. The CYP2B6/RED fusion protein was detected by Western blot, its mRNA by Northern blot, and its heme incorporation into an active form by spectral analysis. Infection with the fusion gene increased RED activity in microsomes by a factor of 3 compared to the control. After infection and treatment with CPA, in cell lines with low endogenous RED, the fusion protein mediated significantly higher CPA-induced cytotoxicity compared to cells expressing solely CYP2B6. In conclusion, the fusion protein is functional for VDEPT by providing one protein for higher levels of CPA metabolism. PMID- 15746947 TI - Dye-assisted small incision cataract surgery in eyes with cataract and coexisting corneal opacity. PMID- 15746948 TI - Drusen classification in bilateral drusen and fellow eye of exudative age-related macular degeneration. AB - AIM: To assess the value of the modified international classification system in screening high-risk patients with bilateral age-related maculopathy (ARM) from those with lower risk characteristics. METHODS: In total, 164 digital images of 106 patients with either bilateral ARM (group A) or the fellow eyes of unilateral exudative age-related macular degeneration (AMD) (Group B) were included. Patients with no signs of ARM in both eyes or those with bilateral late AMD were excluded. The images were randomised and then graded by two masked ophthalmologists based on the modified International Classification of ARM. RESULTS: The interobserver consistency between the two graders was high with a Kappa value of 0.82 (SE 0.34, P<0.0001). There were no significant differences in the distribution of the stages of ARM between the two subgroups. Stage 3 was the most common stage in each group for both graders followed by stage 2a in the bilateral drusen group. Stages 1a, 2a and 2b were equally the next common stage in the fellow eye of chordial neovascularisation group. CONCLUSION: A screening system based on clinical characteristics would be of value in risk prediction in a clinical setting. Type of Drusen alone, as identified by the modified International grading system, may not be reliably predictive in screening for patients who are at high risk of developing choroidal neovascularisation. PMID- 15746949 TI - Partial posterior capsulectomy through an anterior approach: an intraocular lens retaining technique in the management of presumed Propionibacterium acnes endophthalmitis. PMID- 15746950 TI - Metastatic choroidal abscess and choroidal neovascularization in a patient with Staphylococcus aureus renal abscess. PMID- 15746951 TI - Lower eyelid tensometry in younger and older normal subjects. AB - PURPOSE: This study aimed to establish normative and repeatability data for lower eyelid tensometry (LET) using a newly developed eyelid tensometer. METHODS: In this prospective consecutive observational case series, 32 normal adult subjects, comprising 12 younger (aged 29+/-5 years, 6M:6F) and 20 older subjects (aged 74+/ 6 years, 10M:10F), underwent LET. In the younger group, LET was measured for postero-anterior (PA), nasal and temporal displacement. Duplicate measurements were taken and these were repeated on a separate occasion. Associations between repeated measurements and between right and left eyes were determined and the coefficient of repeatability for PA measurements was estimated. Differences in LET between males and females were determined using ANOVA. Only PA tensions, with duplicate measurements, were assessed in the older group, and age effects were determined. RESULTS: Mean temporal tension in the younger group was 7.8+/-2.9 mN (milliNewtons)/mm, while nasal tension was 13.0+/-4.6 mN/mm and PA tension was 11.2+/-5.3 mN/mm. There was a good correlation between interoccasion PA measurements (r=0.82-0.84, P<0.005) and between the right and left eyes (r=0.48, P<0.005). The coefficient of repeatability for PA tension was 1.8 mN/mm. In the younger group, overall eyelid tension (for all directions) for males was significantly higher than for females (ANOVA, P=0.03). For measurements in older subjects on a single occasion, PA tension was 11.1+/-3.5 mN/mm and in younger subjects 12.8+/-4.2 mN/mm (P=0.22). CONCLUSIONS: LET is rapid, repeatable, feasible, and acceptable. PA tension was the most repeatable measurement. Younger males had higher eyelid tension than females, and there was no significant reduction in PA tension with age. PMID- 15746952 TI - Efficacy and side effects of 'swinging eyelid' orbital decompression in Graves' orbitopathy: a proposal for standardized evaluation of diplopia. AB - OBJECTIVES: To evaluate the efficacy and side effects of 'swinging eyelid' orbital decompression in patients with Graves' orbitopathy (GO). To calculate the incidence of postoperative new-onset diplopia (NOD) using a newly proposed scoring system for diplopia. METHODS: We reviewed the clinical data on proptosis, visual acuity, and diplopia in 104 consecutive patients (198 orbits) with GO, who underwent orbital decompression. A combined lateral canthal and inferior fornix incision ('swinging eyelid' approach) was used for removal of the medial wall, the orbital floor and, if indicated, the lateral wall. Indications for surgery were disfiguring/congestive GO (DGO) in 79 patients (149 orbits) and compressive optic neuropathy (CON) in 25 patients (49 orbits). Diplopia was scored according to four grades. In both groups, the incidence of new-onset (continuous) diplopia (NOD), deterioration of diplopia (DOD), and improvement of diplopia (IOD) were calculated, using strictly defined criteria. Our data on NOD were compared to those from other series, after recalculation according to our criteria. RESULTS: The mean proptosis reduction was 4.6 mm (range 0-9.5 mm) after three-wall decompression (95 patients, 180 orbits) vs 3.1 mm (range 0-7 mm) after two-wall decompression (nine patients, 18 orbits). The visual acuity improved in 98% of the patients with CON. In patients with DGO, NOD occurred in 14%. In patients with CON, NOD was not observed, but DOD occurred in 41%. Our data compare favourably to the reported incidence of NOD after either transantral or transnasal decompression. CONCLUSIONS: "Swinging eyelid' orbital decompression is efficacious for proptosis reduction as well as for optic nerve decompression. A scoring system for standardized evaluation of diplopia is proposed. PMID- 15746954 TI - Characteristics and surgical outcomes of paediatric retinal detachment. PMID- 15746955 TI - Virus-induced chalazion. AB - PURPOSE: To investigate a viral etiology in certain chalazia. METHODS: A prospective study over 7.5 years of all newly presenting chalazia associated with diffuse follicular conjunctivitis but without any other aetiological factors. Patients were investigated for ocular or systemic infections by history, physical exam, slit-lamp exam, and/or histology of conjunctival biopsy (including transmission electron microscopy). RESULTS: A total of 27 patients developed follicular conjunctivitis without meibomian gland dysfunction, blepharitis, or sexually transmitted diseases. Evidence for a viral aetiology included: recent systemic viral illness (15/27), recent contact with subjects with chalazia or follicular conjunctivitis (5/27), preauricular lymphadenopathy (4/27), viral corneal disease (4/27), or viral particles by ultrastructure (4/4). CONCLUSIONS: Chalazia may be associated with viral conjunctivitis. Intralesional corticosteroids should be considered with great caution for viral-induced chalazia. PMID- 15746956 TI - Simultaneous bilateral acute angle closure glaucoma following venlafaxine treatment. PMID- 15746957 TI - Immunopathogenesis of conjunctival remodelling in vernal keratoconjunctivitis. AB - PURPOSE: To study the processes involved in mediating conjunctival remodelling in vernal keratoconjunctivitis (VKC) by investigating the expression of integrin receptors, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), transforming growth factor-beta(TGF-beta), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and Ki67 antigen, which is a marker for cell proliferation. METHODS: Conjunctival biopsy specimens from 16 patients with active VKC and nine control subjects were studied by immunohistochemical techniques using monoclonal and polyclonal antibodies directed against the integrin alpha3 and alpha6 subunits, EGFR, VEGF, TGF-beta, bFGF, PDGF, and Ki67 antigen. The phenotype of inflammatory cells expressing growth factors was examined by double immunohistochemistry. RESULTS: In the normal conjunctiva, very weak immunoreactivity was observed for EGFR and VEGF in epithelial cells, and for alpha3 and alpha6 integrin subunits on basal epithelial cells, and on vascular endothelial cells in the upper substantia propria. There was no immunoreactivity for the other antibodies. In VKC specimens, strong staining for alpha3 and alpha6 integrin subunits was observed on the membranes of basal and suprabasal epithelial cells, and all vascular endothelial cells. Immunoreactivity for Ki67 antigen was observed in the nuclei of the basal and suprabasal epithelial cells. Strong immunoreactivity was observed for EGFR in the deeper layers of the epithelium, and for VEGF in all epithelial cells. Inflammatory cells expressing EGFR, VEGF, TGF-beta, bFGF, and PDGF were noted in 8, 9, 11, 10, and 10 specimens, respectively. The majority of inflammatory cells expressing growth factors were eosinophils (45+/-4%) and monocytes/macrophages (35+/-4%). CONCLUSIONS: Chronic conjunctival inflammation in VKC is associated with increased staining of alpha3, and alpha6 integrin subunits, EGFR, VEGF, TGF beta, bFGF, and PDGF that might mediate conjunctival remodelling. PMID- 15746958 TI - Bardet-Biedl syndrome associated with glaucoma. PMID- 15746960 TI - Retinal detachment surgery outside specialist centres. PMID- 15746961 TI - Deep sclerectomy augmented with mitomycin C. AB - AIMS: To investigate the comparative efficacy and safety of deep sclerectomy with and without intraoperative mitomycin C (MMC) application for lowering the intraocular pressure (IOP). METHODS: A total of 71 eyes of 71 consecutive patients who had routine deep sclerectomy (DS), nonaugmented (DS-noMMC) or with mitomycin C (DS-MMC) augmentation (0.2 mg/ml for 2 min) and follow-up of 4 months or more were identified from an ongoing prospective database on glaucoma surgery. Indications for MMC use were the presence of risk factors for subconjunctival scarring and low target IOPs. MMC 0.2 mg/ml was applied in the sub-Tenons space for 2 min. RESULTS: There were 19 eyes in the DS-noMMC group and 52 eyes in the DS-MMC group. In 11 eyes (15.5%), the procedure was complicated by intraoperative perforation of the trabeculo-Descemet's window. Eyes in the DS-MMC group had significantly lower IOPs (MANOVA, P = 0.04). Kaplan-Meier survival curve analysis showed that the probability of maintaining IOP below target IOP level, below 18 mmHg and below 14 mmHg at 1 year was 51, 67, and 35% for the DS-noMMC group and 80, 86, and 74% for the DS-MMC group. The survival rates of the DS-MMC group were not statistically significant (P = 0.06) when the success criterion was maintaining an IOP less than 18 mmHg but were significant for the other criteria, namely IOP less than target levels (P = 0.03) and less than 14 mmHg (P = 0.03). Nd:YAG goniopuncture to lower IOP to target levels was done more frequently in the DS-noMMC group (13 eyes, 81%) than the DS-MMC group (20 eyes, 45%) and this difference was significant (P = 0.03). The prevalence of avascular areas within filtration blebs and transconjunctival oozing of aqueous was significantly higher in the DS-MMC group (P < 0.01). CONCLUSIONS: The use of intraoperative MMC during deep sclerectomy has a significant effect on the postoperative IOP and increases the probability of achieving target IOPs. However, our current technique of MMC application is associated with a higher incidence of avascular blebs and transconjunctival oozing. PMID- 15746962 TI - The Ras-MAPK signal transduction pathway, cancer and chromatin remodeling. AB - Stimulation of the Ras-mitogen-activated protein kinase (MAPK) signal transduction pathway results in a multitude of events including expression of the immediate-early genes, c-fos and c-myc. Downstream targets of this stimulated pathway are the mitogen- and stress-activated protein kinases (MSK) 1 and 2, which are histone H3 kinases. In chromatin immunoprecipitation assays, it has been shown that the mitogen-induced phosphorylated H3 is associated with the immediate-early genes and that MSK1/2 activity and H3 phosphorylation have roles in chromatin remodeling and transcription of these genes. In oncogene-transformed fibroblasts in which the Ras-MAPK pathway is constitutively active, histone H1 and H3 phosphorylation is increased and the chromatin of these cells has a more relaxed structure than the parental cells. In this review we explore the deregulation of the Ras-MAPK pathway in cancer, with an emphasis on breast cancer. We discuss the features of MSK1 and 2 and the impact of a constitutively activated Ras-MAPK pathway on chromatin remodeling and gene expression. PMID- 15746963 TI - Chromatin supraorganization, DNA fragmentation, and cell death in snake erythrocytes. AB - In nucleate erythrocytes of several vertebrate groups, the frequency and intensity of DNA fragmentation associated with programmed cell death vary considerably. Although hemoglobin efficiency may be related to erythrocyte life span, and hemoglobin types and erythrocyte life spans are assumed to vary in reptiles, no data on DNA fragmentation and chromatin organization as related to cell death exist for snakes. In the present study, chromatin supraorganization, DNA fragmentation, and cell death were investigated in four snake species (Crotalus durissus terrificus, Bothrops jararaca, Bothrops alternatus, and Bothrops neuwiedii), which differ in their geographical distribution and habitats, by using image analysis of Feulgen hydrolysis kinetics, the TUNEL assay, single-cell gel electrophoresis, and transmission electron microscopy. Relatively few circulating erythrocytes were found to be simultaneously committed to cell death, although there was some variation among the snake species. Conspicuous nuclear and cytoplasmic organelles suggestive of metabolic activity were seen ultrastructurally in most snake erythrocytes. The DNA of the snake erythrocyte chromatin was much more resistant to Feulgen acid hydrolysis (DNA depurination and breakdown) than that of young adult bullfrog erythrocytes, which had a high frequency and intensity of DNA fragmentation. Of the species studied, B. neuwiedii and C. d. terrificus showed the greatest resistance to Feulgen acid hydrolysis and to the DNA fragmentation, revealed by the TUNEL assay. Although B. neuwiedii also showed the lowest frequency of cells with more damaged DNA in the single-cell gel electrophoresis assay, C. d. terrificus had the highest frequency of damaged cells, possibly because of the abundance of alkaline-sensitive DNA sites. The results for DNA fragmentation and cell death in erythrocytes of B. jararaca and B. alternatus generally differed from those for C. d. terrificus and B. neuwiedii and may reflect differences in the biology of these species selected under different geographical habitats. The differences in erythrocyte cell biology reported here may be related to hemoglobin variants selected in the mentioned snake species and that would lead the cells to different resistances to unfavorable environmental conditions. PMID- 15746964 TI - Angiostatin and plasminogen share binding to endothelial cell surface actin. AB - Previous studies from this laboratory have demonstrated that plasminogen binds to endothelial cell surface-associated actin via its kringles in a dose-dependent and specific manner. The purpose of this study was to determine whether angiostatin, a proteolytic fragment of plasminogen, shares binding properties with plasminogen. Our results indicated that like plasminogen, angiostatin bound to actin in a time-, concentration-, and kringle-dependent manner. Furthermore, this binding was significantly inhibited by excess plasminogen, suggesting that both proteins shared binding motifs on the actin molecule. Fluorescence studies demonstrated that angiostatin bound to intact endothelial cells through its kringles, and this binding was also inhibited by plasminogen but not by unrelated proteins. Ligand blot analyses on endothelial cell lysates indicated that angiostatin interacted with a 42 kDa protein, which was identified as actin. Furthermore, an anti-actin antibody inhibited binding of angiostatin to endothelial cells by approximately 25%. These results suggest that angiostatin and plasminogen share binding to endothelial cell surface actin and, therefore, that angiostatin has the potential to inhibit plasmin-dependent processes such as cell migration-movement. PMID- 15746965 TI - Chemokine receptor CXCR4-beta1 integrin axis mediates tumorigenesis of osteosarcoma HOS cells. AB - It is known that beta1 integrins mediate the migratory response of cells to chemokine stimulation. Also, both beta1 integrins and chemokines have roles in tumor development. In the present study, the beta1 integrin-chemokine axis is assessed using human osteosarcoma (HOS) transfectant cells expressing the CXCR4 receptor for chemokine SDF-1 (CXCL12). We first identified in vitro the specific beta1 integrins that mediated the migratory response to SDF-1 stimulation. Results showed that on collagen type I and laminin, the chemotactic response to SDF-1 was predominantly mediated by alpha2beta1 integrin. On fibronectin, SDF-1 stimulated chemotaxis involved both alpha4beta1 and alpha5beta1 integrins. A comparison of the transfectant clones expressing CXCR4 at low, intermediate, and high levels and the control transfectant revealed that the transfectant clones migratory response in vitro and their ability to form tumors in vivo was related to their levels of CXCR4 expression. In addition, treatment by injection with mAbs to CXCR4, integrin alpha2beta1, or integrin alpha5beta1 effectively inhibited the growth of HOS-CXCR4 transfectant cells in vivo. Therefore, our results show that the beta1 integrins that mediated the migratory response were also functionally linked to the enhanced tumor growth of CXCR4-expressing HOS transfectant cells. PMID- 15746966 TI - Analysis of an approach to oviduct-specific expression of modified chicken lysozyme genes. AB - The -2.7 kb enhancer (E) element of the chicken lysozyme gene domain appears to govern expression of the gene in macrophages but not in oviduct tubular gland cells, the only other site of lysozyme expression. The ultimate goal of our research was to determine whether lysozyme domain variants could be developed that would mainly be expressed in the oviduct so that transgenic birds could be produced that would deposit exogenous protein in the egg white. Accordingly, precise mutations were made by poxvirus-mediated gene targeting in FEF/PU.1 and CCAAT/enhancer-binding protein (C/EBP) transcription factor binding sites in the 2.7 kb E of cloned copies of a specific lysozyme gene variant that includes a hydrophobic pentapeptide tail encoding sequence inserted immediately prior to the stop codon. This variant contains the entire lysozyme domain and is cloned in a lambda bacteriophage vector (lambdaDIILys-HT); the novel tail sequence enables distinction in cell-based expression systems between transcripts of the variant and those of the endogenous gene. These various lysozyme domain mutants, in bacteriophage vector form, were tested for expression in cultured chicken blastodermal cells cotransfected with plasmids encoding the transcription factors C/EBP and v-Myb. In the absence of these plasmids, barely detectable levels of endogenous lysozyme gene transcription resulted in the blastodermal cells. In the presence of the plasmids, however, transcripts of the endogenous gene could be detected as well as varying levels (as evaluated by quantitative real-time PCR) of transcripts of all of the lysozyme domain mutants. These results are discussed in the context of the known role and occurrence of various transcription factors involved in gene expression in differentiating macrophage cells. The ultimate test of expression of the variants in macrophages vs. oviduct cells will be to use them to produce transgenic birds. PMID- 15746967 TI - Structural characterization of the antigenic capsular polysaccharide and lipopolysaccharide O-chain produced by Actinobacillus pleuropneumoniae serotype 15. AB - The specific capsular polysaccharide produced by Actinobacillus pleuropneumoniae serotype 15 was determined to be a high-molecular-mass polymer having [alpha]D + 69 degrees (water) and composed of a linear backbone of phosphate diester linked disaccharide units of 2-acetamido-2-deoxy-D-glucose (D-GlcNAc) and 2-acetamido-2 deoxy-D-galactose (D-GalNAc) residues (1:1). Thirty percent of the D-GalNAc residues were substituted at O-4 by beta-D-galactopyranose (beta-D-Galp) residues. Through the application of chemical and NMR methods, the capsule, which defines the serotype specificity of the bacterium, was found to have the structure [structure: see text]. The O-polysaccharide (O-PS) component of the A. pleuro pneumoniae serotype 15 lipopolysaccharide (LPS) was characterized as a linear unbranched polymer of repeating pentasaccharide units composed of D glucose (2 parts) and D-galactose (3 parts), shown to have the structure [structure: see text]. The O-PS was chemically identical with the O-antigen previously identified in the LPSs produced by A. pleuro pneumoniae serotypes 3 and 8. PMID- 15746968 TI - [Methods of chemical immobilization of an enzyme on a solid support]. AB - Carbonic anhydrase was immobilized on a polymer thermoplastic to promote gaseous CO2 hydration into bicarbonate ions. Catalyst immobilization was realized through a series of chemical reactions enabling enzyme covalent binding to polyamide support. Different initial enzyme concentrations of 0.25, 0.50, 0.75, 1, and 2 mg/ml were verified. Different techniques were developed to assess enzyme attachment. The amount of bound protein was determined using Bradford dosage of proteins remaining in solution following CA II incubation with solid support. ELISA has given a qualitative evaluation of the protein, enabling a follow up of enzyme binding robustness as a function of time. p-nitrophenyl acetate hydrolysis and CO2 hydration were assessed by spectrophotometry and electrometry, respectively. The percentage of active enzyme following immobilization was measured using an esterase linear model. Catalytic transformation rates for gaseous CO2 hydration were calculated for each type of immobilization. PMID- 15746969 TI - Rose (Rosa rugosa)-flower extract increases the activities of antioxidant enzymes and their gene expression and reduces lipid peroxidation. AB - The effects of rose-flower extract on antioxidant enzymes were studied. The activities of catalase (CAT) and glutathione peroxidase (GPx) in 9-month-old senescence-accelerated mice (SAM mice) were lower than those in 6-month-old SAM mice. Therefore, 9-month-old SAM mice were the most appropriate targets for treatment with the rose-flower extract. The activities of CAT and GPx in SAM mice treated with rose-flower extract showed a marked increase in whole blood and liver. At the same time, the gene-expression level of CAT and GPx was upregulated in the liver, while malondialdehyde content in liver and brain decreased. Male SAM mice were more sensitive than female SAM mice. The mean and the longest lifespan of SAM mice were longer after treatment with rose-flower extract. PMID- 15746970 TI - Tocotrienol-induced cytotoxicity is unrelated to mitochondrial stress apoptotic signaling in neoplastic mammary epithelial cells. AB - Tocotrienols and tocopherols represent the 2 subgroups within the vitamin E family of compounds, but tocotrienols display significantly greater apoptotic activity against a variety of cancer cell types. However, the exact mechanism mediating tocotrienol-induced apoptosis is not understood. Studies were conducted to determine the effects of tocotrienols on mitochondrial-stress-mediated apoptotic signaling in neoplastic +SA mammary epithelial cells grown in vitro. Exposure for 24 h to 0-20 micromol/L gamma-tocotrienol resulted in a dose responsive increase in +SA cells undergoing apoptosis, as determined by flow cytometric analysis of Annexin V staining. However, tocotrienol-induced apoptosis was not associated with a disruption or loss of mitochondrial membrane potential, or the release of mitochondrial cytochrome c into the cytoplasm, as determined by JC-1 flow cytometric staining and ELISA assay, respectively. Interestingly, apoptotic +SA cells showed a paradoxical decrease in mitochondrial levels of pro apoptotic proteins Bid, Bax, and Bad, and a corresponding increase in mitochondrial levels of anti-apoptotic proteins, Bcl-2 and Bcl-xL, suggesting that mitochondrial membrane stability and integrity might actually be enhanced for a limited period of time following acute tocotrienol exposure. In summary, these findings clearly demonstrate that tocotrienol-induced apoptosis occurs independently of mitochondrial stress apoptotic signaling in neoplastic +SA mammary epithelial cells. PMID- 15746971 TI - Influence of mechanical and biological signals on gene expression in human MG-63 cells: evidence for a complex interplay between hydrostatic compression and vitamin D3 or TGF-beta1 on MMP-1 and MMP-3 mRNA levels. AB - Biological mediators can influence the activity and differentiation of bone cells. 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3) is known to induce differentiation of precursors into mature osteoblasts, and transforming growth factor-beta1 (TGF-beta1) can modulate the activity of bone cells leading to alterations in proliferation and gene expression patterns. Bone-derived cells were loaded via intermittent cyclic hydrostatic pressure (icHP) on cells under basal conditions and in the presence of 1,25-(OH)2D3 or TGF-beta1. Evaluating the effects of loading on the cells allowed for a comparison to be made between responsiveness to biomechanical and biochemical stimuli and their potential interplay. The effects of icHP on mRNA levels for the specific genes involved in bone remodelling and differentiation were measured in MG-63 cells using reverse transcription-polymerase chain reaction (RT-PCR). The mRNA levels for matrix metalloproteinase-1 and -3 (MMP-1 and MMP-3) were significantly, and uniquely, increased (p < 0.001) in cells exposed to icHP under serum-free conditions for 4 12 h. However, mRNA levels for MMP-3, but not MMP-1, were significantly enhanced in cells subjected to static hydrostatic pressure (HP). Treatment of cells with 1,25-(OH)2D3 resulted in increased (p < 0.001) mRNA levels for osteocalcin and decreased (p < 0.001) mRNA levels for both MMP-1 and MMP-3. In cells exposed to icHP and 1,25-(OH)2D3, the mRNA levels for both MMP-1 and MMP-3 were elevated (p < 0.001) compared with hormone alone, but not to the same degree (p < 0.01) as cells subjected to icHP alone. Addition of TGF-beta1 to cells led to increases in cell proliferation and expression of collagen I, as well as decreases in expression of osteocalcin and MMP-1 and MMP-3. Exposure of cells to icHP and TGF beta1 again led to unique and significant increases in expression of MMP-1 and MMP-3. No changes in mRNA levels for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or any of the other 9 genes assessed, including those for MMP-2 and MMP 13, were detected under any of the conditions described. Therefore, icHP can induce alterations in mRNA levels for a specific subset of genes in both premature and mature osteoblasts. Such stimuli can modulate the impact of potent biological mediators in defining patterns of gene expression by bone cells and potentially modify function in vivo. PMID- 15746972 TI - Approaches to biology teaching and learning: from a scholarly approach to teaching to the scholarship of teaching. PMID- 15746973 TI - From the National Academies: medical school admissions requirements and undergraduate science education. PMID- 15746974 TI - Video views and reviews: cytokinesis: a phenomenon overlooked too often. PMID- 15746975 TI - Meeting report: synthetic biology Jamboree for undergraduates. PMID- 15746976 TI - Meeting report: building bridges for diverse professors of tomorrow. PMID- 15746977 TI - Points of view: effective partnerships between K-12 and higher education. PMID- 15746978 TI - Probabilities and predictions: modeling the development of scientific problem solving skills. AB - The IMMEX (Interactive Multi-Media Exercises) Web-based problem set platform enables the online delivery of complex, multimedia simulations, the rapid collection of student performance data, and has already been used in several genetic simulations. The next step is the use of these data to understand and improve student learning in a formative manner. This article describes the development of probabilistic models of undergraduate student problem solving in molecular genetics that detailed the spectrum of strategies students used when problem solving, and how the strategic approaches evolved with experience. The actions of 776 university sophomore biology majors from three molecular biology lecture courses were recorded and analyzed. Each of six simulations were first grouped by artificial neural network clustering to provide individual performance measures, and then sequences of these performances were probabilistically modeled by hidden Markov modeling to provide measures of progress. The models showed that students with different initial problem-solving abilities choose different strategies. Initial and final strategies varied across different sections of the same course and were not strongly correlated with other achievement measures. In contrast to previous studies, we observed no significant gender differences. We suggest that instructor interventions based on early student performances with these simulations may assist students to recognize effective and efficient problem-solving strategies and enhance learning. PMID- 15746979 TI - Developing information fluency in introductory biology students in the context of an investigative laboratory. AB - Students of biology must learn the scientific method for generating information in the field. Concurrently, they should learn how information is reported and accessed. We developed a progressive set of exercises for the undergraduate introductory biology laboratory that combine these objectives. Pre- and postassessments of approximately 100 students suggest that increases occurred, some statistically significant, in the number of students using various library related resources, in the numbers and confidence level of students using various technologies, and in the numbers and confidence levels of students involved in various activities related to the scientific method. Following this course, students should be better prepared for more advanced and independent study. PMID- 15746980 TI - Understanding of genetic information in higher secondary students in northeast India and the implications for genetics education. AB - Since the work of Watson and Crick in the mid-1950s, the science of genetics has become increasingly molecular. The development of recombinant DNA technologies by the agricultural and pharmaceutical industries led to the introduction of genetically modified organisms (GMOs). By the end of the twentieth century, reports of animal cloning and recent completion of the Human Genome Project (HGP), as well techniques developed for DNA fingerprinting, gene therapy and others, raised important ethical and social issues about the applications of such technologies. For citizens to understand these issues, appropriate genetics education is needed in schools. A good foundation in genetics also requires knowledge and understanding of topics such as structure and function of cells, cell division, and reproduction. Studies at the international level report poor understanding by students of genetics and genetic technologies, with widespread misconceptions at various levels. Similar studies were nearly absent in India. In this study, I examine Indian higher secondary students' understanding of genetic information related to cells and transmission of genetic information during reproduction. Although preliminary in nature, the results provide cause for concern over the status of genetics education in India. The nature of students' conceptual understandings and possible reasons for the observed lack of understanding are discussed. PMID- 15746981 TI - Nonreplication in genetic studies of complex diseases--lessons learned from studies of osteoporosis and tentative remedies. AB - Inconsistent results have accumulated in genetic studies of complex diseases/traits over the past decade. Using osteoporosis as an example, we address major potential factors for the nonreplication results and propose some potential remedies. Over the past decade, numerous linkage and association studies have been performed to search for genes predisposing to complex human diseases. However, relatively little success has been achieved, and inconsistent results have accumulated. We argue that those nonreplication results are not unexpected, given the complicated nature of complex diseases and a number of confounding factors. In this article, based on our experience in genetic studies of osteoporosis, we discuss major potential factors for the inconsistent results and propose some potential remedies. We believe that one of the main reasons for this lack of reproducibility is overinterpretation of nominally significant results from studies with insufficient statistical power. We indicate that the power of a study is not only influenced by the sample size, but also by genetic heterogeneity, the extent and degree of linkage disequilibrium (LD) between the markers tested and the causal variants, and the allele frequency differences between them. We also discuss the effects of other confounding factors, including population stratification, phenotype difference, genotype and phenotype quality control, multiple testing, and genuine biological differences. In addition, we note that with low statistical power, even a "replicated" finding is still likely to be a false positive. We believe that with rigorous control of study design and interpretation of different outcomes, inconsistency will be largely reduced, and the chances of successfully revealing genetic components of complex diseases will be greatly improved. PMID- 15746982 TI - PGE2 signal through EP2 promotes the growth of articular chondrocytes. AB - EP2 was identified as the major PGE2 receptor expressed in articular cartilage. An EP2 agonist increased intracellular cAMP in articular chondrocytes, stimulating DNA synthesis in both monolayer and 3D cultures. Hence, the EP2 agonist may be a potent therapeutic agent for degenerative cartilage diseases. INTRODUCTION: Prostaglandin E2 (PGE2) exhibits pleiotropic effects in various types of tissue through four types of receptors, EP1-4. We examined the expression of EPs and effects of agonists for each EP on articular chondrocytes. MATERIALS AND METHODS: The expression of each EP in articular chondrocytes was examined by immunohistochemistry and RT-PCR. A chondrocyte cell line, MMA2, was established from articular cartilage of p53(-/-) mice and used to analyze the effects of agonists for each EP. A search for molecules downstream of the PGE2 signal through the EP2 agonist was made by cDNA microarray analysis. The growth promoting effect of the EP2 agonist on chondrocytes surrounded by cartilage matrix was examined in an organ culture of rat femora. RESULTS AND CONCLUSION: EP2 was identified as the major EP expressed in articular cartilage. Treatment of MMA2 cells with specific agonists for each EP showed that only the EP2 agonist significantly increased intracellular cAMP levels in a dose-dependent manner. Gene expression profiling of MMA2 revealed a set of genes upregulated by the EP2 agonist, including several growth-promoting and apoptosis-protecting genes such as the cyclin D1, fibronectin, integrin alpha5, AP2alpha, and 14-3-3gamma genes. The upregulation of these genes by the EP2 agonist was confirmed in human articular chondrocytes by quantitative mRNA analysis. On treatment with the EP2 agonist, human articular chondrocytes showed an increase in the incorporation of 5-bromo-2-deoxyuracil (BrdU), and the organ culture of rat femora showed an increase of proliferating cell nuclear antigen (PCNA) staining in articular chondrocytes surrounded by cartilage matrix, suggesting growth-promoting effects of the PGE2 signal through EP2 in articular cartilage. These results suggested that the PGE2 signal through EP2 enhances the growth of articular chondrocytes, and the EP2 agonist is a candidate for a new therapeutic compound for the treatment of degenerative cartilage diseases. PMID- 15746983 TI - Influence of glucocorticoids on human osteoclast generation and activity. AB - Using human peripheral blood mononuclear cells as osteoclast precursors, we showed that dexamethasone stimulated osteoclast generation at a pharmacological concentration but did not affect the life span of human osteoclasts. Dexamethasone also dose-dependently increased signals for osteoclastogenesis. INTRODUCTION: Glucocorticoid-induced osteoporosis is a common and serious disease. Glucocorticoids predominantly affect osteoblast proliferation and life span. Much of the bone loss is caused by reduced bone formation, but there is also an element of increased bone resorption. MATERIALS AND METHODS: Human peripheral blood mononuclear cells were cultured on whale dentine and induced to differentiate to osteoclasts by RANKL and human macrophage-colony stimulating factor (M-CSF). Osteoclast activity was quantified by pit area. RANKL and osteoprotegerin (OPG) expression in osteoblasts were measured by real-time RT PCR. RESULTS: In the early phase of osteoclast generation (0-16 days), cultures from two different donors showed that dexamethasone at 10(-8) M increased pit area by 2.5-fold, whereas lower concentrations had no effect. At the highest dexamethasone concentration (10(-7) M), pit area was reduced. In 21-day cultures from three other donors, a similar increase was seen with dexamethasone at 10(-8) M. There was, however, no evidence of increased life span of osteoclasts with dexamethasone. In human primary osteoblasts, dexamethasone dose-dependently reduced OPG and increased RANKL expression as measured by quantitative real time RT-PCR. CONCLUSION: These data provide some explanation at a cellular and molecular level for the observed increase in bone resorption seen in patients treated with glucocorticoids and indicate that there are clear direct effects of glucocorticoids on bone resorption in human cell systems that may differ from other species. PMID- 15746984 TI - Alveolar bone marrow as a cell source for regenerative medicine: differences between alveolar and iliac bone marrow stromal cells. AB - We isolated and expanded BMSCs from human alveolar/jaw bone at a high success rate (70%). These cells had potent osteogenic potential in vitro and in vivo, although their chondrogenic and adipogenic potential was less than that of iliac cells. INTRODUCTION: Human bone marrow stromal cells (BMSCs) have osteogenic, chondrogenic, and adipogenic potential, but marrow aspiration from iliac crest is an invasive procedure. Alveolar BMSCs may be more useful for regenerative medicine, because the marrow can be aspirated from alveolar bone with minimal pain. MATERIALS AND METHODS: In this study, alveolar bone marrow samples were obtained from 41 patients, 6-66 years of age, during the course of oral surgery. BMSCs were seeded and maintained in culture with 10% FBS and basic fibroblast growth factor. In addition, BMSCs were induced to differentiate into osteoblasts, chondrocytes, or adipocytes in appropriate medium. RESULTS AND CONCLUSION: From a small volume (0.1-3 ml) of aspirates, alveolar BMSCs expanded at a success ratio of 29/41 (70%). The success rate decreased with increasing donor age, perhaps because of age-dependent decreases in the number and proliferative capacity of BMSCs. The expanded BMSCs differentiated into osteoblasts under osteogenic conditions in 21-28 days: the mRNA levels of osteocalcin, osteopontin, and bone sialoprotein, along with the calcium level, in alveolar BMSC cultures were similar to those in iliac cultures. However, unlike iliac BMSC, alveolar BMSC showed poor chondrogenic or adipogenic potential, and similar differences were observed between canine alveolar and iliac BMSCs. Subsequently, human alveolar BMSCs attached to beta-tricalcium phosphate were transplanted into immunodeficient mice. In transplants, new bone formed with osteoblasts and osteocytes that expressed human vimentin, human osteocalcin, and human GAPDH. These findings suggest that BMSCs have distinctive features depending on their in vivo location and that alveolar BMSCs will be useful in cell therapy for bone diseases. PMID- 15746985 TI - Azidothymidine induces apoptosis and inhibits cell growth and telomerase activity of human parathyroid cancer cells in culture. AB - Telomerase activity has been correlated to parathyroid carcinoma. Because its role in acquisition of a malignant phenotype by parathyroid cells is unclear, we treated telomerase-positive cultured human parathyroid cancer cells with the telomerase inhibitor AZT, evaluating cell telomerase activity, cytotoxic effects, growth, and morphological changes. In vitro exposure of these cells to AZT correlated with inhibition of cell proliferation. INTRODUCTION: Parathyroid carcinoma represents an uncommon cause of primary hyperparathyroidism, whose spectrum of clinical presentation, degree of malignancy, and prognosis are difficult to be properly identified. Neck surgery, specifically an en bloc resection of primary tumor, is the only curative treatment. Alternatively, affected patients could undergo repetitive palliative surgical exeresis of metastatic nodules. It has been previously shown that telomerase activity is specifically present in parathyroid carcinoma cells, being absent in hyperplastic and adenomatous tissues. Thus, determination of telomerase activity could represent either a useful diagnostic molecular marker for human parathyroid carcinoma or a potential target for pharmacological intervention in a malignant neoplasia usually resistant to chemo- and radiotherapeutic interventions. MATERIALS AND METHODS: To further investigate the role of telomerase activity in acquisition of a malignant phenotype by parathyroid cells, we treated telomeric repeat amplification protocol-positive cultured human parathyroid cells with the telomerase inhibitor zidovudine, 3'-azido-3'deoxythymidine (AZT), evaluating cell telomerase activity, growth characteristics, potential cytotoxic effects, and morphological changes. RESULTS: Our findings indicate that in vitro exposure of human parathyroid cancer cells to AZT resulted in intracellular accumulation of AZT-monophosphate (AZT-MP) and inhibition of telomerase, which correlate with inhibition of human parathyroid cancer cell proliferation. Moreover, we also found that AZT induced an apoptotic rather than a necrotic type of cellular death. None of these effects were observed in human adenomatous parathyroid cells in culture. CONCLUSIONS: Altogether these results indicate that AZT may be a highly effective agent against cancer parathyroid cells proliferation, which is an extremely important observation for a neoplasia which shows lack of response to classical pharmacological and physical antiblastic treatments. PMID- 15746986 TI - Impaired marrow osteogenesis is associated with reduced endocortical bone formation but does not impair periosteal bone formation in long bones of SAMP6 mice. AB - We used the SAMP6 osteoporotic mouse to examine the link between marrow osteogenic potential and in vivo cortical bone formation. SAMP6 marrow supported less in vitro osteogenesis than marrow from SAMR1 controls; SAMP6 mice had a corresponding deficit in endocortical mineralizing surface. This marrow/endocortical defect did not affect the periosteum, where SAMP6 mice had normal to enhanced bone formation. INTRODUCTION: With aging, there may be a reduction in the number or proliferative capacity of bone marrow osteoprogenitors that may contribute to age-related decreases in bone formation. To examine the link between the ability of the marrow to support osteogenesis and age-related changes in bone formation, we measured in vitro and in vivo indices of osteogenesis in a model of osteoporosis, the senescence-accelerated mouse SAMP6. MATERIALS AND METHODS: Femora and tibias from SAMP6 and SAMR1 (control) mice were harvested at 2, 4, 6, and 12 months of age (168 bones total). Bone marrow cells were cultured under osteogenic conditions and stained for alkaline phosphatase (ALP) and alizarin red. Dynamic indices of bone formation were assessed histologically from calcein labels. RESULTS: ALP+ and alizarin red-positive areas were significantly less in cultures from SAMP6 bones versus SAMR1 (p < 0.05), indicating less osteogenic potential. For example, SAMP6 tibial cultures had 21% less ALP+ area and 36% less alizarin red-positive area than SAMR1. Marrow from tibias had 2-fold greater osteogenesis than femoral marrow (p < 0.001). SAMP6 mice had a deficit in endocortical mineralizing surface across all age groups (p < 0.05), but no deficit in mineral apposition rate. Last, despite the marrow and endocortical deficits, SAMP6 mice had normal or slightly increased periosteal bone formation, consistent with their larger bone size. CONCLUSION: SAMP6 bone marrow supports less in vitro osteogenesis than SAMR1, consistent with a lower concentration of marrow osteoprogenitors in SAMP6. SAMP6 mice have less endocortical mineralizing surface than SAMR1 at all ages but no detectable deficit in mineral apposition rate, which suggests a reduction in osteoblast number but normal function. Periosteal bone formation is unimpaired in SAMP6 mice, indicating that the marrow/endocortical defect does not affect the periosteal surface. PMID- 15746987 TI - Expression of Runx2/Cbfa1/Pebp2alphaA during angiogenesis in postnatal rodent and fetal human orofacial tissues. AB - Transient expression of Runx2 is reported in endothelial cells and vascular smooth muscle cells during vessel formation in skin, stroma of forming bones and developing periodontal ligament, developing skeletal muscle cells, and fat tissue. The data suggest that Runx2 is expressed in a multipotential mesenchymal cell population that gives rise to various osseous and nonosseous cell lineages. INTRODUCTION: Runx2/Cbfa1 is a transcription factor essential for cells of osteogenic and dentinogenic lineages. Here we examined expression of Runx2/Cbfa1 (all isotypes) in several nonskeletal cell types present in developing orofacial tissues of neonatal rodents and human fetuses with special emphasis on vessel formation. MATERIALS AND METHODS: Sections obtained from heads or jaws of postnatal mice, hamster, and human fetuses were immunostained with monoclonal anti-Pebp2aA antibody. Mouse and human tissues were also examined by in situ hybridization. Sections of Runx2 null mutant mice with a LacZ reporter construct inserted in the Runx2 locus were stained for Runx2 promoter activity with anti galactosidase. RESULTS: We found transient mRNA and protein expression in endothelial cells and in vascular smooth muscle cells of forming vessels in skin, alveoli of forming bone, and forming periodontal ligament. We also noticed weak and variable expression in some fibroblasts of embryonic skin, early differentiating cross-striated muscle cells, and differentiating fat cells. CONCLUSION: Runx2 is not an exclusive marker for chondrogenic, osteogenic, and dentinogenic tissues, but is much more widely present in an early multipotential mesenchymal cell population that gives rise to several other lineages. PMID- 15746988 TI - Mechanotransduction in bone does not require a functional cyclooxygenase-2 (COX 2) gene. AB - COX-2 is a key enzyme involved in the response of bone to loading. However, using mice with a null mutation of the COX-2 gene, we found that a functional COX-2 gene is not required for mechanotransduction. This paradoxical finding may have resulted, in part, from mechanically induced COX-1 activity. INTRODUCTION: Cyclooxygenase-2 (COX-2) is an important mediator in the response of bone to mechanical loading, with pharmacological inhibition of COX-2 effectively eliminating or reducing mechanically induced bone formation. In this study, we further investigated the role of COX-2 in skeletal mechanotransduction using a genetic approach. The aim was to compare the skeletal responsiveness of COX-2 homozygous mutant (COX-2(-/-)) and wildtype control (COX-2(+/+)) mice to investigate whether a functional COX-2 gene is necessary for mechanotransduction. MATERIALS AND METHODS: Adult female COX-2(+/+) and COX-2(-/-) mice on a C57BL/6x129/ola background were studied using the ulna axial loading model. The response to 2 days of loading for 120 cycles/day at 2 Hz was measured histomorphometrically. Phenotypic characterization of the femurs in these mice was also performed. In a separate group of animals, the expression of the remaining COX isozyme, COX-1, was assessed using real-time RT-PCR 4 h after one bout of 120 loading cycles. RESULTS: Null mutation of the COX-2 gene resulted in a consistent femoral phenotype of reduced bone mass, altered architecture, and inferior mechanical properties. Many of these differences were nullified after adjustment for body weight. Nevertheless, body weight-corrected values showed a consistent trend of reduced mechanical properties in COX-2(-/-) mice. Genotype did not influence the response to mechanical loading, with no histomorphometric differences being found between COX-2(+/+) and COX-2(-/-) mice. Real-time RT-PCR showed COX-2(-/-) mice to express significantly greater COX-1 expression in loaded ulnas than in loaded ulnas in COX-2(+/+) mice. There were no differences in COX-1 expression in nonloaded ulnas. CONCLUSIONS: A functional COX-2 gene was not found to be required for skeletal mechanotransduction. This is in contrast to previous pharmacological studies showing that COX-2 is critical to the response of bone to loading. Investigating a potential reason for the absence of a genotype difference in this study, we found that mice with a null mutation in the COX-2 gene possess inductive skeletal COX-1 expression. PMID- 15746989 TI - Mechanism and function of high vitamin D receptor levels in genetic hypercalciuric stone-forming rats. AB - The functional status and mechanism of increased VDR in GHS rats were investigated. Basal VDR and calbindins were increased in GHS rats. 1,25(OH)(2)D(3) increased VDR and calbindins in controls but not GHS rats. VDR half-life was prolonged in GHS rats. This study supports the mechanism and functional status of elevated VDR in GHS rats. INTRODUCTION: Genetic hypercalciuric stone-forming (GHS) rats form calcium kidney stones from hypercalciuria arising from increased intestinal calcium absorption and bone resorption and decreased renal calcium reabsorption. Normal serum 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] levels and increased vitamin D receptor (VDR) protein suggest that high rates of expression of vitamin D-responsive genes may mediate the hypercalciuria. The mechanism of elevated VDR and state of receptor function are not known. MATERIALS AND METHODS: GHS and non-stone-forming control (NC) male rats (mean, 249 g), fed a normal calcium diet, were injected intraperitoneally with 1,25(OH)2D3 (30 ng/100 g BW) or vehicle 24 h before cycloheximide (6 mg/100 g, IP) and were killed 0-8 h afterward. Duodenal VDR was measured by ELISA and Western blot, and duodenal and kidney calbindins (9 and 28 kDa) were measured by Western blots. RESULTS AND CONCLUSIONS: Duodenal VDR protein by Western blot was increased 2-fold in GHS versus NC rats (633 +/- 62 versus 388 +/- 48 fmol/mg protein, n = 4, p < 0.02), and 1,25(OH)2D3 increased VDR and calbindins (9 and 28 kDa) further in NC but not GHS rats. Duodenal VDR half-life was prolonged in GHS rats (2.59 +/- 0.2 versus 1.81 +/- 0.2 h, p < 0.001). 1,25(OH)2D3 prolonged duodenal VDR half-life in NC rats to that of untreated GHS rats (2.59 +/- 0.2 versus 2.83 +/- 0.3 h, not significant). This study supports the hypothesis that prolongation of VDR half-life increases VDR tissue levels and mediates increased VDR-regulated genes that result in hypercalciuria through actions on vitamin D-regulated calcium transport in intestine, bone, and kidney. PMID- 15746990 TI - Overweight postmenopausal women lose bone with moderate weight reduction and 1 g/day calcium intake. AB - Overweight postmenopausal women may be more susceptible to bone loss with weight reduction than previously studied obese women. The influence of energy restriction and Ca intake on BMD was assessed in 66 individuals. Weight reduction resulted in bone loss at several sites in women consuming 1 g Ca/day and was mitigated with higher calcium intake at 1.7 g/day. INTRODUCTION: Bone loss is associated with weight loss in obese postmenopausal women and can be prevented with calcium (Ca) supplementation. However, because bone loss caused by weight loss may be greater in overweight than obese women, it is not clear whether Ca supplementation is also beneficial in overweight women. MATERIALS AND METHODS: We assessed the influence of caloric restriction at two levels of Ca intake on BMD and BMC in 66 overweight postmenopausal women (age, 61 +/- 6 years; body mass index, 27.0 +/- 1.8 kg/m2). Subjects completed either a 6-month energy-restricted diet (WL, n = 47) and lost 9.3 +/- 3.9 % weight or maintained weight (WM; 1 g Ca/day, n = 19). Participants in the WL group were randomly assigned to either normal (1 g/day; WL NL-Ca) or high (1.7 g/day; WL Hi-Ca) Ca intake. Regional BMD and BMC were measured at baseline and after 6 months. RESULTS: During normal Ca intake, trochanter BMD and BMC and total spine BMD were decreased more in WL than WM women (p < 0.05). The WL NL-Ca group lost more trochanter BMD (-4.2 +/- 4.1%) and BMC (-4.8 +/- 7.1%) than the WL Hi-Ca group (-1.4 +/- 5.6% and -1.1 +/- 8.1%, respectively; p < 0.05). There were no significant changes in BMD or BMC at the femoral neck in any group. Weight loss correlated with trochanter BMD loss (r = 0.687, p < 0.001) in the WL NL-Ca group. CONCLUSION: Despite an intake of 1 g Ca/day, bone loss occurred at some sites because of weight loss. Calcium intake of 1.7 g/day will minimize bone loss during weight loss in postmenopausal overweight women. PMID- 15746991 TI - Effects of low-dose prednisone on bone metabolism. AB - Prednisone 5 mg/day suppresses multiple indices of bone formation in a randomized placebo-controlled trial in healthy postmenopausal females. This suggests that even low doses of prednisone may reduce bone repair or renewal and may have adverse effects on bone mass and/or bone strength. INTRODUCTION: High doses of chronic glucocorticoids are known to have adverse effects on bone, and measures to prevent bone loss are well established for doses >7.5 mg daily, because these doses can cause premature or exaggerated osteoporosis. However, it is unclear if chronic prednisone doses of 5 mg daily have the same effects on bone. There are no established recommendations for preventing glucocorticoid-induced osteoporosis in people taking prednisone 5 mg daily, a dose used frequently in medical practice to treat diseases of the lungs, joints, skin, muscles, eyes, nerves, etc. Our primary objective was to test whether prednisone 5 mg daily affects serum and urine indices of bone metabolism in healthy postmenopausal women. Our secondary objectives were to determine if prednisone 5 mg affected systolic or diastolic blood pressure or causes side effects. MATERIALS AND METHODS: A double blinded randomized placebo-controlled 8-week trial in 50 healthy postmenopausal women was conducted at the Massachusetts General Hospital Outpatient General Clinical Research Center. Patients were randomly assigned to prednisone 5 mg daily or matching placebo for 6 weeks, followed by a 2-week recovery phase. Markers of bone formation and resorption were determined at weeks 0, 2, 4, 6, and 8. Indices of osteoblast activity included serum propeptide of type I N-terminal procollagen (PINP), propeptide of type I C-terminal procollagen (PICP), osteocalcin, and bone-specific alkaline phosphatase (BSALP). Indices of osteoclast activity included urine and serum type I collagen N-telopeptide (NTX) and free urinary deoxypyridinoline (DPD). RESULTS AND CONCLUSIONS: Prednisone rapidly and significantly decreased serum PINP (p < 0.01), PICP (p < 0.01), and osteocalcin (p < 0.01) and free urinary deoxypyridinoline (p = 0.017). These changes were largely reversed during the recovery period. Side effects were indistinguishable in the two groups. Neither systolic nor diastolic blood pressure changed significantly throughout the study between the two groups. In conclusion, low-dose prednisone significantly decreases indices of bone formation and may decrease indices of bone resorption in postmenopausal women. Further studies are needed to assess the effects of low-dose prednisone on BMD and fracture risk. PMID- 15746992 TI - L-arginine, the natural precursor of NO, is not effective for preventing bone loss in postmenopausal women. AB - NO is an important regulator of bone turnover. L-Arginine, the natural precursor of NO, can enhance NO production. However, no effect of L-arginine hydrochloride supplementation was found on bone metabolism or on BMD, bone mass, or bone structure of healthy postmenopausal women. INTRODUCTION: Recent studies indicate that NO exerts an anabolic effect on bone cell activity. The NO level of the human body can be elevated by administering pharmacological NO donors. Animal studies and the first human trial showed that NO donor administration had a positive effect on bone formation and a negative effect on bone resorption. L arginine, the natural precursor of NO, can enhance NO production. This study was conducted to examine the effect of an oral L-arginine supplement on bone metabolism of healthy postmenopausal women. MATERIALS AND METHODS: The participants in this study were 30 healthy, age-matched postmenopausal women, divided into two groups. For 6 months, one group (54.5 +/- 4.1 years; 66.3 +/- 10.5 kg) received a daily oral supplement with 18 g L-arginine hydrochloride (14.8 g free L-arginine). The other 15 volunteers (55.3 +/- 4.4 years; 64.2 +/- 9.1 kg) received 18 g dextrose as a placebo. To verify compliance, 24-h urinary excretion of nitrogen was analyzed for 2 consecutive days at baseline and after 2, 4, and 6 months. At baseline and after 2, 4, and 6 months of supplementation, blood was drawn for analysis of insulin-like growth factor-I (IGF-I) and biomarkers of bone metabolism. At baseline, after 6 months, and after 1 year, pQCT measurements were performed at trabecular and cortical sites of the radius and tibia. The two groups of subjects were compared by repeated measures ANOVA. RESULTS: As expected, in the group with L-arginine hydrochloride supplementation, nitrogen excretion rose, and in the placebo group, it remained constant. Only bone formation marker, procollagen type I propeptides (PICP), increased significantly (p < 0.05) after 6 months of L-arginine supplementation. The results from pQCT showed no significant changes at any site in either group. No significant change in IGF-I concentration, which might have been caused by the L arginine hydrochloride supplementation, was evident. CONCLUSIONS: We conclude from these results that supplementation with L-arginine hydrochloride is not effective for improving bone mass in humans. PMID- 15746993 TI - Recombinant human TSH modulates in vivo C-telopeptides of type-1 collagen and bone alkaline phosphatase, but not osteoprotegerin production in postmenopausal women monitored for differentiated thyroid carcinoma. AB - In women monitored for thyroid carcinoma, short-term stimulation with rhTSH induced an acute decrease in serum C-telopeptides of type-1 collagen and an increase in serum BALP levels without any effect on OPG production. The inhibitory effect of TSH on bone resorption occurred only in postmenopausal women who showed low BMD and a high bone turnover rate as an effect of L-thyroxine suppressive therapy. INTRODUCTION: It has been recently shown that thyrotropin (TSH) has an inhibitory activity on skeletal remodeling in in vitro conditions. Here, we have aimed at evaluating whether TSH has similar effects in vivo. For this purpose, we have evaluated the sequential profile of serum bone metabolism markers during acute stimulation with recombinant human TSH (rhTSH) in thyroidectomized women monitored for thyroid carcinoma. MATERIALS AND METHODS: The study group included 66 thyroidectomized patients, of whom 38 were premenopausal and 28 postmenopausal, who underwent routine rhTSH-assisted whole body radioactive iodine scanning for differentiated thyroid carcinoma. The patients were sequentially evaluated for TSH, free triiodothyronine (FT3), free thyroxine (FT4), bone alkaline phosphatase (BALP), C-telopeptides of type-1 collagen (CrossLaps), and osteoprotegerin (OPG) levels during rhTSH stimulation. The samples were drawn just before and 2 and 7 days after the first administration of rhTSH. BMD was evaluated by ultrasonography at baseline. Seventy-one healthy women (41 premenopausal and 30 postmenopausal) acted as a control group. RESULTS AND CONCLUSIONS: At study entry, all patients had subclinical thyrotoxicosis as effect of L-thyroxine (L-T4) treatment. The patients had higher serum CrossLaps and OPG levels and lower BMD than healthy subjects. Postmenopausal patients showed comparable serum FT4 and FT3 concentrations with those found in premenopausal patients. However, postmenopausal patients showed higher serum CrossLaps (p < 0.001), OPG (p = 0.03), and BALP (p < 0.001) levels and lower BMD (p < 0.001) than those measured in premenopausal patients. Two days after the first administration of rhTSH, all patients had serum TSH values >100 mUI/liter. At this time, serum CrossLaps levels decreased significantly (p < 0.001) and BALP values increased (p = 0.001) with respect to the baseline values in postmenopausal but not in premenopausal patients. rhTSH did not induce any significant change in serum OPG values either in premenopausal or in postmenopausal patients. One week after the first rhTSH administration, serum CrossLaps values decreased again to values comparable with those measured at baseline, whereas serum BALP values remained high. This study shows that subclinical thyrotoxicosis is accompanied by high bone turnover rate with an increase in serum OPG levels compared with euthyroid healthy subjects. Acute increase in serum TSH levels is accompanied by a reversible inhibition of bone resorption. This effect is characterized by a decrease in serum CrossLaps and an increase in BALP levels without any evident effect on OPG production. The activity of TSH occurs specifically in postmenopausal women in whom the negative effects of L-T4 suppressive therapy on bone mass and metabolism are more marked compared with premenopausal women. PMID- 15746994 TI - Fracture risk with multiple myeloma: a population-based study. AB - Pathologic fractures, especially of the axial skeleton, are extremely common in patients with multiple myeloma and cluster around the time of diagnosis. Osteoporotic fractures seem to be less of a problem in these patients. INTRODUCTION: It is generally believed that fractures are common in patients with multiple myeloma as a result of lytic bone lesions, generalized bone loss, and/or elevated bone turnover from excessive cytokine production, but the actual risk of pathologic versus osteoporotic fractures has not been quantified. MATERIALS AND METHODS: In a population-based retrospective cohort study, 165 Olmsted County, MN, residents with myeloma diagnosed from 1945 to 2001 (55% men; mean age, 70.7 +/- 11.1 years) were followed for 537 person-years. The relative risk of fractures was assessed by standardized incidence ratios (SIRs), and risk factors were evaluated in proportional hazards models. RESULTS: Altogether, 134 patients experienced 463 fractures. In the year before diagnosis, 16 times more fractures were observed than expected, mostly pathologic fractures of the vertebrae and ribs. Subsequently, there was a 9-fold increase in fracture risk. However, 69% of these fractures were pathologic, and another 11% were found incidentally on myeloma monitoring. With the latter two groups excluded, subsequent fracture risk was elevated 3-fold, with a 2-fold increase in the risk of an osteoporotic fracture. In multivariate analyses, the predictors of overall fracture risk were oral corticosteroid use and elevated serum calcium levels, whereas pathologic fractures were additionally predicted by use of chemotherapy. CONCLUSION: There is a dramatic increase in fractures around the time of diagnosis of myeloma, most of which are pathologic fractures. The most important predictor of overall fracture risk is oral corticosteroid use. PMID- 15746995 TI - Development and initial validation of a risk score for predicting in-hospital and 1-year mortality in patients with hip fractures. AB - Our objectives were to better define the rates and determinants of in-hospital and 1-year mortality after hip fracture. We studied a population-based cohort of 3981 hip fracture patients. Using multivariable regression methods, we identified risk factors for mortality (older age, male sex, long-term care residence, 10 prefracture co-morbidities) and calculated a hip fracture-specific score that could accurately predict or risk-adjust in-hospital and 1-year mortality. Our methods, after further validation, may be useful for comparing outcomes across hospitals or regions. INTRODUCTION: Hip fractures in the elderly are common and associated with significant mortality and variations in outcome. The rates and determinants of mortality after hip fracture are not well defined. Our objectives were (1) to define the rate of in-hospital and 1-year mortality in hip fracture patients, (2) to describe co-morbidities at the time of fracture, and (3) to develop and validate a multivariable risk-adjustment model for mortality. MATERIALS AND METHODS: We studied a population-based cohort of 3981 hip fracture patients > or =60 years of age admitted to hospitals in a large Canadian health region from 1994 to 2000. We collected sociodemographic and prefracture co morbidity data. Main outcomes were in-hospital and 1-year mortality. We used multivariable regression methods to first derive a risk-adjustment model for mortality in 2187 patients treated at one hospital and then validated it in 1794 patients treated at another hospital. These models were used to calculate a score that could predict or risk-adjust in-hospital and 1-year mortality after hip fracture. RESULTS AND CONCLUSIONS: The median age of the cohort was 82 years, 71% were female, and 26% had more than four prefracture co-morbidities. In-hospital mortality was 6.3%; 10.2% for men and 4.7% for women (adjusted odds ratio, 1.8; 95% CI, 1.3-2.4). Mortality at 1 year was 30.8%; 37.5% for men and 28.2% for women (adjusted p < 0.001). Older age, male sex, long-term care residence, and 10 different co-morbidities were independently associated with mortality. Risk adjustment models based on these variables had excellent accuracy for predicting mortality in-hospital (c-statistic = 0.82) and at 1 year (c-statistic = 0.74). We conclude that 1 in 15 elderly patients with hip fracture will die during hospitalization, and almost one-third of those who survive to discharge will die within the year. The determinants of mortality were primarily older age, male sex, and prefracture co-morbidities. Our hip fracture-specific risk-adjustment tool is pragmatic and reliable, and after further validation, may be useful for comparing outcomes across different hospitals or regions. PMID- 15746996 TI - Interobserver reproducibility of criteria for vertebral body exclusion. AB - We studied reproducibility of the ISCD vertebral exclusion criteria among four interpreters. Surprisingly, agreement among interpreters was only moderate, because of differences in threshold for diagnosing focal structural defects and choice of which vertebra among a pair discordant for T-score, area, or BMC to exclude. Our results suggest that reproducibility may be improved by specifically addressing the sources of interobserver disagreement. INTRODUCTION: Although DXA is widely used to measure vertebral BMD, its interpretation is subject to multiple confounders including osteoarthritis, aortic calcification, and scoliosis. In an attempt to standardize interpretation and minimize the impact of artifacts, the International Society for Clinical Densitometry (ISCD) established criteria for vertebral exclusion, including the presence of a focal structural defect (FSD), discrepancy of >1 SD in T-score between adjacent vertebrae, and a lack of increase in BMC or area from L1 to L4. Whereas the efforts of the ISCD represent an important advance in BMD interpretation, the interobserver reproducibility with application of these criteria is unknown. We hypothesized that there would be substantial agreement among four interpreters regarding application of the exclusion criteria and the final lumbar spine T-score. MATERIALS AND METHODS: Each interpreter read a set of 200 lumbar DXA scans obtained on male veterans, applying the ISCD vertebral body exclusion criteria. RESULTS: Surprisingly, agreement among interpreters was only moderate. Differences in interpretation resulted from differing thresholds for recognition of FSD and the choice of excluding the upper or lower vertebral body for the criteria requiring comparison between adjacent vertebrae. CONCLUSIONS: Despite their apparent simplicity, the ISCD vertebral exclusion criteria are difficult to apply consistently. In principle, appropriate refinement of the exclusion criteria may significantly improve interobserver agreement. PMID- 15746997 TI - Initial years of recreational artistic gymnastics training improves lumbar spine bone mineral accrual in 4- to 8-year-old females. AB - Gymnasts' bone mineral characteristics are generally not known before starting their sport. Prepubertal females who enrolled in beginning artistic gymnastics (n = 65) had lower bone mineral than controls (n = 78). However, 2 years of gymnastics participation versus no participation led to a significantly greater accrual of forearm bone area and lumbar spine areal BMD. INTRODUCTION: The skeletal response to exercise in children compared with adults is heightened because of the high bone turnover rate and the ability of bone to change its size and shape. Whereas child gymnasts generally have greater rates of bone mineral accrual compared with nongymnasts, it is unknown if some of these skeletal advantages are present before the onset of training or are caused entirely by training. MATERIALS AND METHODS: Changes in bone area (BA; cm2), BMC (g), and areal BMD (aBMD; g/cm2) over 24 months were examined in prepubertal females, 4-8 years of age, who selected to perform recreational gymnastics (GYM; n = 65), nongymnastic activities, or no organized activity (CON; n = 78). Participants had essentially no lifetime history of organized athletic participation (< 12 weeks). Pubertal maturation was assessed annually by a physician. Total body, lumbar spine, total proximal femur, and forearm BA, BMC, and aBMD were measured every 6 months using DXA (Hologic QDR-1000W). Independent samples t-tests determined baseline group differences. Nonlinear mixed effects models were used to model 24 month changes in bone data. In subset analyses, high-level gymnasts advancing to competition (HLG; n = 9) were compared with low-level nonadvancing gymnasts (LLG; n = 56). RESULTS: At baseline, GYM were shorter, lighter, and had lower BA, BMC, and aBMD compared with CON (p < 0.05), whereas HLG did not differ significantly in these measurements compared with LLG (p > 0.05). Controlling for differences in race, baseline measures of body mass, height, and calcium intake, and change in breast development beyond stage II at 24 months, GYM had greater long-term (asymptotic) mean responses for total body aBMD and forearm BMC (p < 0.04) and greater rates of increase in the mean responses of lumbar spine aBMD and forearm BA compared with CON over 24 months. Over time, forearm BA increased to a greater extent in HLG compared with LLG (p < 0.01). CONCLUSIONS: Females participating in recreational gymnastics initiated during childhood have enhanced bone mineral gains at the total body, lumbar spine, and forearm over 24 months. Higher-level training promotes additional gains in forearm BA. PMID- 15746998 TI - Femoral neck structure in adult female athletes subjected to different loading modalities. AB - Loading modality is a strong external determinant of structure and concomitant strength of the femoral neck. Particularly effective seem to be loadings, which arise from high impacts or impacts from atypical loading directions. INTRODUCTION: Physical loading plays an important role, not only in the process of bone modeling and remodeling, but also in shaping a mechanically appropriate bone structure. This study aimed at testing the hypothesis that the type of loading partly determines the femoral neck structure. MATERIALS AND METHODS: A total of 255 premenopausal female athletes representing volleyball, hurdling, squash-playing, soccer, speed skating, step aerobics, weight-lifting, orienteering, cross-country skiing, cycling, and swimming and their 30 nonathletic counterparts were measured with DXA. Besides the conventional areal BMD (aBMD) of the femoral neck, the hip structure analysis (HSA) was used to estimate the cross-sectional area (CSA), subperiosteal width (W), and section modulus (Z, an index of bone strength) at the narrowest section of the femoral neck. Also, training history, muscle strength, and calcium intake were assessed. The above-mentioned sports were classified according to the type of loading they apparently produce at the hip region; that is, high-impact loading (volleyball, hurdling), odd-impact loading (squash-playing, soccer, speed-skating, step aerobics), high-magnitude loading (weightlifting), low-impact loading (orienteering, cross-country skiing), and nonimpact loading (swimming, cycling). RESULTS: High-impact and odd-impact loading sports were associated with the highest age-, weight-, and height-adjusted aBMD (23% and 29% higher values compared to nonathletic referents), CSA (22% and 27%), and Z (22% and 26%). In contrast, repetitive, nonimpact loading sports were not associated with any clear benefit in any bone value compared with the referents. The W at the narrowest femoral neck section was similar in all groups. Body height and weight accounted virtually for one-half of the variation in Z, whereas the type of loading predicted 13% of the total variation of this variable-clearly more than the 2% attributable to isometric leg extension strength. Both high-impact and odd-impact loading modalities were associated with a large benefit in Z, corresponding to >1 SD in the reference group, whereas repetitive, low-impact loading showed a benefit that was only one-half of that. Surprisingly, high-magnitude loading (weightlifting) was not associated with a statistically significant increase in Z. CONCLUSION: We conclude that mechanical loading and its modality are strong external determinants of structure and concomitant strength of the femoral neck. Particularly effective seem to be loadings, which arise from high impacts or impacts from atypical loading directions. PMID- 15746999 TI - Males have larger skeletal size and bone mass than females, despite comparable body size. AB - Gender differences in fractures may be related to body size, bone size, geometry, or density. We studied this in 18-year-old males (n = 36) and females (n = 36) matched for height and weight. Despite comparable body size, males have greater BMC and BMD at the hip and distal tibia and greater tibial cortical thickness. This may confer greater skeletal integrity in males. INTRODUCTION: Gender differences in fractures may be related to body size, bone size, geometry, or density. We studied this in males (n = 36) and females (n = 36; mean age = 18 years) pair-matched for height and weight. MATERIALS AND METHODS: BMC, bone area (BA), and BMD were measured in the spine and hip using DXA. Distal tibia was measured by pQCT. RESULTS AND CONCLUSIONS: Males had a higher lean mass (92%) compared with females (79%). No gender differences were observed for vertebral BMC or vertebral height, although males had greater width and thus BA at the spine. Males had greater BMC and BA at the femoral neck and total femur (p < 0.02). Geometric variables of the hip including neck diameter and neck-axis length were also greater in males (p < 0.02). There was greater cross-sectional moment of inertia, safety factor, and fall index in males (all p < 0.02). Males had greater tibial BMC, volumetric BMD, and cortical area and thickness compared with females (p < 0.01), with both greater periosteal circumference (p = 0.011) and smaller endosteal circumference (p = 0.058). Statistically controlling for lean mass reduced gender differences, but males still had 8% higher hip BMD (p = 0.24) and 5.3% higher total tibial BMD (p = 0.05). A subset of males and females were matched (n = 14 pairs) for total hip BA. Males in this subset still had greater BMC and BMD at the total hip (p < 0.05) than females, despite similar BA. In summary, despite comparable body size, males have greater BMC and BMD than females at the hip and distal tibia but not at the spine. Differences in BMC and BMD were related to greater cortical thickness in the tibia. We conclude that differences in bone mass and geometry confer greater skeletal integrity in males, which may contribute to the lower incidence of stress and osteoporotic fractures in males. PMID- 15747002 TI - [Facts on higher quality papers and journals]. PMID- 15747000 TI - Risk assessment and fracture discrimination by ultrasound: the debate continues. PMID- 15747003 TI - [Clinical evaluation of nickel-titanium rotary instruments Hero 642 in root canal preparation]. AB - PURPOSE: This research is aimed to assess the clinical value of the Nickel titanium (NiTi) rotary instruments Hero 642 in root canal preparation. METHODS: 60 molars were instrumented by Hero 642 rotary instruments using crown-down technique in the experimental group, and by K files using step-back technique in the control group. All teeth were obturated with lateral condensation method. The efficiency of preparation and obturation was analyzed by radiographs before and after treatment. The canal curvature and operation time were compared by means of Student's t test, and the incidence of complications and post-treatment pain were compared by means of Chi square test between two groups. RESULTS: No transportation, apical blockage, ledge or perforation was found in the experimental group. The Hero 642 instruments could keep the original curvature and flow of the root canals. There was more canal complications in the control group than in the experimental group (P<0.005). The operative time was shorter and post treatment pain seldom occurred in the experimental group. CONCLUSION: The NiTi rotary instruments Hero 642 can be used for preparation of root canals effectively and safely, and is worth wide application. PMID- 15747004 TI - [A study of multi-factors on the utilization of dental services in Chinese residents]. AB - PURPOSE: In this article, multi-factors on the utilization of dental services was assessed in Chinese residents by Logistic regression model. METHODS: A household oral health interview survey was conducted among Chinese residents. A study of multi-factors on the utilization of dental services in Chinese residents was analyzed by Logistic regression model. RESULTS: The data 1558 residents in 587 houses had been collected from 6 sampling spots by the household oral health interview survey. Significant difference was found in three hypothesis test of Logistic regression model over 15 year old of residents. CONCLUSIONS: The utilization of dental services was decided by multi-factors in Chinese residents. The main factors to influence residents visiting dentist were dental price, living area and dental caries. PMID- 15747005 TI - [Short-term observation of gold-plating unprecious metal ceramic crown in restoring anterior tooth]. AB - PURPOSE: To evaluate the short-term effect of gold-plating technique used in Ni Cr metal ceramic crown in restoring anterior teeth. METHODS: 64 teeth from 38 cases were divided into two groups. Each group consisted of 32 teeth. All the teeth were restored with Ni-Cr metal ceramic crown. The restorations in group 1 had no treatment while those in group 2 were treated with gold-plating technique in the base metal alloy.Gingiva discoloration, bleeding on probing (BOP), plaque index (PLI) and sulcular bleeding index (SBI) were examined 1 and 12 months after restoration. Chi-square test and Wilcoxon rank sum test were used for statistical analysis. RESULTS: Gingiva discoloration was found in both groups after bonding. According to the percentage of gingival discoloration, there was no significant difference between group 1 and group 2 (12.5% vs 28.1%) (P>0.05). However, 12 months later, significant difference was found between the two groups (15.6% vs 37.5%)(P<0.05). According to PLI, BOP and SBI, there was no statistical difference between two groups (P>0.05). CONCLUSION: Gold-plating was a good method in solving the problem of gingival discoloration in unprecious metal ceramic crown. PMID- 15747006 TI - [Application of X ray cephalogram in the measurement of adenoids in children]. AB - PURPOSE: To evaluate the clinical value of X ray lateral cephalogram in the measurement of adenoids in children. METHODS: 45 cases (aged from 3 to 13 year old) with adenoid hypertrophy suspected clinically were examined with lateral cephalometric projections, of which 40 cases were examined with lateral nasopharyngeal projections at one time. Then the quality of films were appraisal and the adenoids were measured on the film. Student's X(2) test was used for statistics analysis. RESULTS: X ray lateral cephalogram can distinctly reveal the structure of nasopharynx. The method was simply and reproducible. The quality of the films were determined based on the conjunction between the base of the pterygoid plate and extracranial aspect of the occipital slope, with consideration of the mandibular margin and sphenoid saddle. The conjunction should be clearly demonstrated and the edges of the mandibular margin and sphenoid saddle should be sharp and well demarcated in qualified films. 45 cases were examined with lateral cephalometric projections, 34 cases had standard films, accounting for 76%. 40 cases were examined with lateral nasopharyngeal projections, 21 had standard films, accounting for 53%. The quality of X ray lateral cephalogram was significantly better than lateral nasopharyngeal projections (P<0.05). CONCLUSION: Compared with the routine lateral nasopharyngeal projection, lateral nasopharyngeal cephalogram has images of high quality, is better for showing the nasopharyngeal structures as well as measurement of the adenoids with parenchyma. It is the imaging method of choice for children with OSAHS. PMID- 15747007 TI - [Investigation of extracellular signal-regulated protein kinase and lymphnode metastasis in oral squamous cell carcinoma]. AB - PURPOSE: To study the relationship between the expression of extracellular signal regulated protein kinase (ERK) and the differentiation degree and lymphnode metastasis in oral squamous cell carcinoma. METHODS: Immunohistochemical staining was used in 41 cases of OSCC to investigate the expression of ERK protein. Student's t test were used for statistical analysis. RESULTS: A significant relationship was observed between the expression intensity of ERK protein and clinicopathological features of OSCC (P<0.01). A significant relationship was observed between the expression intensity of ERK protein and lymph node metastasis of OSCC(P<0.05). No correlation between the expression intensity of ERK protein and cell differentiation degree of OSCC (P>0.05) was found. CONCLUSIONS: Activation of ERK may play a strong role in the development of OSCC. Intensive expression of ERK protein may be an important presage marker in OSCC patients. PMID- 15747009 TI - [Evaluation of the clinical effect of the teeth with subgingivally involved defect conserved by crown lengthening surgery]. AB - PURPOSE: To evaluate the clinical effect of the teeth with subgingivally involved defect which were conserved by crown lengthening surgery. METHODS: 62 teeth, with defect subgingivally from 1.5 mm to 4 mm, mobility degree(MD)0.05), but a significant increase about MD occurred in the major defect group one year after restoration (P<0.01), and there was significant correlation between MD of each stage after operation and PD of pre-operation in anterior teeth (r=0.489, 0.526, 0.531, P<0.01). CONCLUSIONS: According to the biological width principle, crown lengthening surgery may conserve these teeth with subgingivally involved defect, and has a good, long time clinical effect. But MD showed an increasing trend after operation and significant cor. PMID- 15747008 TI - [Retention and rehabilitation of complex maxillofacial defects based on bar-clip and stud attachment]. AB - PURPOSE: To evaluate the application of precise attachment in restoration of complicated maxillofacial defect. METHODS: A framework with bar-clip and stud attachment was designed and fabricated. Maxillofacial prosthesis with this framework was applied to treat a patient with defects of nose, entire upper lip and the corresponding maxillofacial areas. The retention force was tested with Universal Testing Device. RESULTS: After treatment, the patient got a better appearance, normal function of mastication and clear pronunciation. The dislodgement test showed the retention force was 40N. CONCLUSION: Application of precise attachment in complicated maxillofacial defects can get satisfactory result. PMID- 15747010 TI - [Histological location of DMP1, OPN and BSP in cementum formation of mouse]. AB - PURPOSE: To study the distribution of dentin matrix protein 1, osteopontin and bone sialoprotein during cementum formation and cementoblasts differentiation in mouse. METHODS: Thirty six BALB/c postnatal mice were divided into four groups according to the developing stages, nine in each was subdivided into three parts randomly to examine the location of DMP1, OPN and BSP, a tooth developing study model was built and examined using histological methods correspondingly. PV two steps immunohistochemical assay was used to localize the distribution of the three regulatory factors timely and spatially. Repeated measures ANOVA and statistical analysis software of CMIAS were used to analyze the intensity of the dyed images. RESULTS: There was a series expression of the three regulatory proteins. DMP1 was expressed in the dental follicle cells in day 5 of postnatal stages and negatively expressed when the cell began to differentiate; meanwhile, since day 10 and on OPN was positively located in cementoblasts until the root developed completely; but BSP was expressed in the same cell in day 15 to 20, and then decreased to be negative. There were significant differences statistically among the three factors in each stage observed in the study except OPN between day 15 and 25. CONCLUSIONS: There are different locations timely and spatially among the three proteins, it hints that they play important different roles during the differentiation of cementoblasts and formation of cementum. PMID- 15747011 TI - [The morphology of the cells in the cartilage of rabbit mandibular condyle]. AB - PURPOSE: To study the tissue layers and their function of the cartilage in mandibular condyle in rabbits. METHODS: Six adult Japanese white rabbits were subjected. Their temporomandibular joints were studied by immunohistochemistry for FGFR3 and PCNA, and in situ hybridization for aggrecan and collagen II mRNA expression, as well as ultrastructure. RESULTS: The upper proliferative cells did not express FGFR3, but the lower proliferative cells expressed FGFR3. Only few cells in the upper proliferative layer were PCNA positive, but all cells in the lower proliferative layer were positive for PCNA. No collagen II mRNA expression was found in the upper proliferative cell, but aggrecan and collagen II mRNA coexpressed in the lower proliferative layer. The cells in both layers were different in ultrastructure. CONCLUSION: The cartilage in mandibular condyle should have the 5 following tissue layer: fibrous layer, proliferative layer, transitional layer, cartilaginous layer and calcified cartilaginous layer. The cells in the proliferative layer are undifferentiated and the cells in the differentiated layer are prechondrocytes. PMID- 15747012 TI - [The influence of autogenous bone graft in cleft palate on maxillary growth: an experiment study in rats]. AB - PURPOSE: This study was designed to investigate the influence of autogenous bone graft in cleft palate on maxillary growth in rats. METHODS: Forty Wistar strain male rats aged 4 weeks were randomly divided into four groups. Each group had 10 rats. Group 1 was the unoperated control group. Group 2 had a surgically created cleft palate but unrepaired. Group 3 also had a surgically created cleft palate which followed by a two-flap palatoplasty repair immediately. Group 4(experimental group) also had a surgically created cleft palate, but the cleft palate was reconstructed by autogenous rib graft with two-flap repair. All animals were sacrificed on the 10th week after operation. The data were analyzed using SPSS. And one-way ANOVA was used for comparison of the four groups. RESULTS: The experimental group was significantly smaller than unoperated control group and cleft palate group in the maxillary widths variables, length variables (AD),height variables(P<0.01).Except rats in the experimental group had significantly greater posterior palate width than rats in two-flap repair group(P<0.01),there were no significant difference between the two procedures in maxillary growth variables(P>0.05).But the experimental group was smaller in the maxillary heights, length variables than the two-flap group. CONCLUSIONS: These findings showed that repair of the cleft palate by autogenous rib graft had positive effect on preventing the contraction of maxillary width. But in the maxillary anteroposterior and vertical development, the experimental group possibly had more severe retardation than two-flap repair group. PMID- 15747013 TI - [Effects of Pingyangmycin albumin microspheres on sclerostenosis of the rabbit central auricular arteries]. AB - PURPOSE: The aim of this study was to investigate the mechanism of Pingyangmycin albumin microspheres (PYM-AMS) acted on the rabbit central auricular arteries. METHODS: In the study of randomized block design, 24 Japanese white rabbits were divided into 4 groups, 6 rabbits in each group. The animals were put on the operation table after being anaesthetized by intramuscular injection of Xumianxin (0.2 ml/Kg) and the interior division was blocked to stop drug liquid get into the interior ear. After being sterilized with 75% alcohol, No.1(PYM hydrochloride injection+0.9% NaCl), No.2(PYM+soybean oil), No.3(PYM-AMS+soybean oil) liquids which contained PYM 5mg/ml were injected into the central auricular arteries of the animals about 30 seconds (0.26 ml/per ear), respectively. Then these vessels were examined histologically after 2,7, 14, 21 days respectively. RESULTS: After injection, in PYM+0.9% NS control group (No.1 liquid), the ears and vessels had no significant changes. In PYM+soybean oil control group (No.2 liquid), at the 2nd day, the endothelial cells were mild swollen. At the 7th day, some endothelial cells were dropped off. At the 14th day, the central auricular arteries had mild change, but the blood flow was not blocked. At the 21st day, the wall of the central auricular artery had more layers, especially on the intima, but the lumen was still obvious. A few of endothelial cells were proliferative. In PYM-AMS group(No.3 liquid), the blood was not resumed at once, and there was some oil in the vessels. At the 1st day, the injection site was mild swollen, and small thrombosis was observed in small vessels. At the 7th-14th day, the central auricular arteries were narrowed, but the blood could be observed. At the 21st day, the vessels had sclerostenosis, and the blood flow was blocked, but no scar and necrosis were observed. Under light microscopy, at the 2nd day, the endothelial cells were mild swollen and small vessels were embolized by PYM-AMS. At the 7th day, the endothelial cells were mild swollen. At the 14th day, the endothelial cells were proliferative and the wall of the central auricular artery had more layers. The lumen of the central artery changed mildly, while the surface of PYM-AMS was absorbed. At the 21st day, the wall of the central auricular artery was proliferative obviously and the artery became sclerostenosed, while cell division was observed. The PYM-AMS was obviously absorbed and some artery was sclerostenosed, while the wall of small vein was proliferative, too. The proliferative intima was proved to be muscle cells with Masson stain and van Gieson's stain. There were many smooth muscle cells, fibroblasts, myofibroblasts in the original blood vascular lumen on the 21st day under electron microscopy. CONCLUSION: Pingyangmycin nonspecifically made the endothelia and vessels injured, and induced proliferation of endothelial cells and smooth muscle cells, and gradually the vessels became sclerostenosed. PYM-AMS had both sustained-releasing effect and embolization effect. PYM-AMS may be a better drug for treatment of large venous malformations and arteriovenous malformations. PMID- 15747014 TI - [The sealing effects of three desensitizers on tubule:an SEM investigation]. AB - PURPOSE: The purpose of this in vitro study was to evaluate the sealing effects on tubule by three different dentin desensitizers. METHODS: 48 freshly extracted human premolars were selected. Their buccal dentin was exposed and a standardized circular area was isolated. They were randomly divided into three experiment groups and one control group. Dentin desensitizers included Systemp desensitizer, Seal&protect desensitizer and Gluma desensitizer. Twelve teeth were prepared for each test group (36 teeth total), and the other twelve teeth were selected as a control group. After the removal of cement, the dentin surfaces were cleaned, treated. Then the teeth were vertically cleaved into two sections. The surface and section of these teeth were observed by means of a scanning electron microscope. RESULTS: All 3 desensitizers could seal the tubules on dentin surface. Systemp desensitizer's dentin permeability was better than Seal&protect, but Gluma had no dentin permeability. CONCLUSION: 3 desensitizers had sealing effect on tubule, systemo desensitizer was the best one. PMID- 15747015 TI - [The expression of hTERT and p53 protein in pre-malignant and malignant lesions of human oral mucosa]. AB - PURPOSE: To investigate the roles of hTERT and p53 protein in malignant changes of oral mucosal precancerous lesions and the relations between hTERT and p53. METHODS: The expression of hTERTmRNA and p53 were measured using in situ hybridization and immunohistochemical assay in 9 cases of oral mucosal hyperplasia, 11 cases of light dysplasia, 10 cases of medium dysplasia, 9 cases of carcinomas in situ and 11 cases of squamous cell carcinomas. The results were processed by medical photographic system, and analyzed statistically by one-way ANOVA. The relation to hTERT and p53 was analyzed statistically by Pearson correlations. RESULTS: It was found that from hyperplasia to light dysplasia, medium dysplasia, carcinomas in situ and squamous cell carcinomas, the indexes of positive cell vessels and the value of OD increased, arriving at the apex in squamous cell carcinoma. With phenotypic progression and the degree of dysplasia, hTERT and p53 expression cells extend from basal layer to the keratinous layer. In highly-differentiated squamous cell carcinomas, expression cells were mainly located around the nests. Stained cells dispersed throughout the tumor tissue in low-differentiated squamous cell carcinomas. p53 was undetectable in hyperplasia and light dysplasia, but p53 was observed in 66.67% cases of high dysplasia and 72.72% cases of squamous cell carcinomas. There was not a significant linear correlation between hTERT and p53. CONCLUSIONS: The results suggest that activation of telomerase and p53 play a role in the process of malignant changes of oral mucosal precancerous lesions. Oral squamous cell carcinoma is a disease that has been found in association with many factors. PMID- 15747016 TI - [Anti-proliferation effect of genistein on salivary adenoid cystic carcinoma cell line SACC-83 in vitro]. AB - PURPOSE: To investigate the anti-proliferation effect of tyrosine protein kinase inhibitor, genistein (4'-5,7-trihydroxyisoflavone), on human salivary adenoid cystic carcinoma cell line SACC-83 in vitro, and its effect on cell cycle. METHODS: SACC-83 cells cultured in vitro were treated with genistein,cell survival rate was calculated with MTT assay, phase contrast microscope was used to observe the status of cell growth and the morphological changes in each group, and cell cycle was detected with flow cytometry. Apoptosis was detected with Annexin V/PI staining. The results were statistically analyzed by SPSS11.5 software. RESULTS: When treated with genistein of certain concentration for certain time, SACC-83 cell growths were significantly inhibited. With the increase of concentration of genistein and elongation of acting time, the inhibitory effects increased. Cells treated with genistein showed changes in morphology, cells' volume was decreased ,and suspended cells increased. Treated with 220 micromol/L genistein for 72 hours, SACC-83 cell growths were significantly inhibited, cell cycle was arrested in G2/M phase, and cell apoptosis was induced(P<0.01). CONCLUSION: Genistein inhibits growth of human salivary adenoid cystic carcinoma cell line SACC-83, arrests cell cycle in G2/M phase, and induces cell apoptosis; these suggest that tyrosine protein kinase has important effect on the neoplasia and development of salivary adenoid cystic carcinoma. PMID- 15747017 TI - [The effect of gemcitabine on proliferation and telomerase activity of Tca8113 cell lines]. AB - PURPOSE: To investigate the effect of gemcitabine on the proliferation and telomerase activity of Tca8113 cell line cells. METHODS: MTT assay, flow cytometry were used to examine the effect of gemcitabine on cell proliferation and cell cycle respectively, telomerase activity was detected with telomeric repeat amplification protocol (TRAP) and polyacrylamide gel electrophoresis (PAGE). The data were analyzed by independent samples t test and one-way ANOVA of SPSS10.0 for windows. RESULTS: Cell proliferations were significantly inhibited in these cells after exposure to gemcitabine as well as cell cycles were arrested. Furthermore, the effect was closely associated with the concentration and time of gemcitabine. The average rate of cell proliferating inhibition in group 5.0 microg/ml at 72 h was 72.2%.The number of cells in S phase was decreased,the average rate of S phase in group 5.0 microg/ml at 72 h was 18.1%,which was lower than that in group 0 microg/ml(38.5%) (t=5.801,P<0.05).Longer exposure time and higher concentration to gemcitabine, much telomerase activity was inhibited. After 72h, the A values of group 0 and 5.0 microg/ml were 1.32+/-0.12, 0.28+/-0.02, respectively (F=811.208, P<0.05). CONCLUSIONS: Gemcitabine could significantly inhibit Tca8113 proliferation and decrease its telomerase activity. Gemcitabine may be applied in clinical therapy of oral cancer. PMID- 15747018 TI - [Expression of Interleukin-8 in KB cells infected with Candida albicans]. AB - PURPOSE: To investigate the change of interleukin-8 (IL-8) in oral epithelial cells (KB cells) infected with Candida albicans. METHODS: Two-site ELISA and RT PCR were applied to determine the levels of IL-8 protein or mRNA expression in the KB cells induced by hyphae, yeast, killed Candida albicans and medium respectively. The unpaired Student's t test was used to analyse the data. RESULTS: Candida albicans induced the KB cells secrete more IL-8. The hyphae, yeast, killed Candida albicans and control group marks of RT-PCR were 180.23, 186.36, 143.41 and 85.57 respectively, calculated using a two-tailed test, P<0.05, was considered to be significant, but the medium group (mark 80.87) had no significant change. The tendency of the IL-8 protein secreted by KB cells increased with the time but reduced when Candida albicans was more. CONCLUSIONS: Candida albicans is responsible for the production of IL-8 secreted by oral epithelial cells which can be expected to play an important role in the infection and defection with the local oral mucosa. PMID- 15747019 TI - [In vitro study of the effect of 11 kinds of natural drugs on the growth and acid production of Lactobacillus]. AB - PURPOSE: To study the effect of 11 different natural drugs on the growth and acid production of Lactobacillus, as a preparation for screening an effective agent to mediate the balance of oral microflora. METHODS: Lactobacillus AC413 was chosen as the experimental bacterium. Eleven kinds of drugs, such as Rhizoma Chuanxiong, Surgentodoxa cuneata and Galla Chinensis were extracted by means of maceration, percolation and reflux extraction. The values of MIC of various extracts were measured. Then, different experimental media containing various extracts were prepared. The concentration of the extracts was lower than the MIC of the drug and the initial pH of the medium was 7.4. Lactobacillus was cultured in the medium for 48 hours, and finally the rest pH was measured. One-way ANOVA was used for statistical analysis. RESULTS: When the concentration of the drugs was lower than 8.000mg/ml, Tea polyphenols, Catechu, Galla Chinensis, Radix et Rhizoma Rhei and Nidus Vespae can inhibit the growth of Lactobacillus effectively. Tea polyphenols, Nidus Vespae, Radix Scuteilariae, Galla Chinensis and Surgentodoxa cuneata can inhibit the acid production of Lactobacillus effectively, Rhizoma Chuanxiong, Radix et Rhizoma Rhei, Semen Arecae and Catechu have no preliminary effect on it, but Surgentodoxa Cuneata and Radix Angelicae Pubescentis can increase it. CONCLUSION: Tea polyphenols, Catechu, Galla Chinensis, Radix et Rhizoma Rhei and Nidus Vespae can inhibit the growth of Lactobacillus effectively, and Tea polyphenols, Radix et Rhizoma Rhei, Nidus Vespae, Radix Scuteilariae, Galla Chinensis and Surgentodoxa Cuneata can inhibit the acid production of Lactobacillus effectively. PMID- 15747020 TI - [A comparison of the activities of membrane-bound, proton translocating ATPases between Streptococcus mutans fluoride-resistant and their parent strains]. AB - PURPOSE: To figure out the reason for increased acid tolerance of Ingbritt-FR, the fluoride-resistant strain of Streptococcus mutans Ingbritt, by determining and comparing the H(+)-ATPase activities of both fluoride-resistant and their parental strains. METHODS: The permeabilized cells of S. mutans Ingbritt and Ingbritt-FR were prepared by treating them with 10% toluene and then two cycles of freezing and thawing. The permeabilized cells were used for ATPase assay by adding them to the reaction mixture which contained 50mM Tris-maleate buffer (pH 6.0), 10 mM MgSO4 and 5 mM ATP. ATPase activity was assessed by measuring inorganic phosphate released from ATP hydrolysis.Two-way ANOVA was used for statistical analysis. RESULTS: The activities of H(+)-ATPase of Ingbritt-FR were 308.48, 136.67, and 82.80 micromol Pi/g cell dry weight/min, at 10, 20, and 60 minutes respectively, significantly higher than those of their parent strain: 104.77, 64.69, and 30.7 (P<0.01). The enzyme activities were decreasing with time. CONCLUSIONS: The higher ATPase activity of fluoride-resistant mutant of S. mutans Ingbritt may account for the increased acid tolerance of this organism, and the increment of ATPase activity and acid tolerance of fluoride-resistant strain is likely to increase the cariogenic potential of S. mutans after fluoride resistant mutation. PMID- 15747022 TI - [Proliferation and ALPase activity of human dental pulp cells in response to lipopolysaccharide in vitro]. AB - PURPOSE: To investigate the effects of LPS on proliferation and ALPase activity of human dental pulp cells (HPCs). METHODS: HPCs were cultivated from connective tissue explants digested with collagenase type I. The expression of mCD14 was determined, and the effects of LPS on proliferation of HPCs at the concentrations of 0.1MUg/ml,1MUg/ml and 10MUg/ml were observed by FCM. The effects of LPS tested on ALPase activity were examined. X(2) test and one-way ANOVA were used in statistical analysis respectively. RESULTS: The results showed that LPS receptor mCD14 was not present in HPCs. LPS inhibited proliferation of HPCs (P<0.01) and ALPase activity in HPCs (P<0.05) at the concentrations of 0.1MUg/ml,1MUg/ml and 10MUg/ml. CONCLUSIONS: It is concluded that LPS tested inhibits proliferation of HPCs and ALPase activity of HPCs. PMID- 15747021 TI - [The antimicrobial effects of chitosans on oral pathogenic microbes in vitro]. AB - PURPOSE: To study the antimicrobial activities of chitosans of different molecular weights and concentrations on oral pathogenic microbes under pH 6.5 in vitro. METHODS: The inhibition effects of chitosans of different molecular weights and different concentrations (2.0%, 1.5%, 1.0%, 0.5%) on Staphylococcus aureus, Candida albicans, Helicobacter pylori, Streptococcus mutans and Porphyromonas gingivalis in culture were investigated, and the differences of their antimicrobial activities under high temperature and after filtering were compared. One way analysis of variance, randomized block design analysis of variance and t test were used for statistical analysis. RESULTS: Chitosans of different molecular weights all showed bacteriostatic effects on Staphylococcus aureus, Candida albicans, Helicobacter pylori, Streptococcus mutans and Porphyromonas gingivalis. Low molecular weight chitosans showed the strongest effects, but the high molecular weight ones had strong effects on Streptococcus mutans (P<0.05,0.01). The bacteriostatic effects of chitosans were not affected by high temperatures (P>0.05). CONCLUSION: Chitosans have inhibitory effects on oral pathogenic microbes under pH 6.5. PMID- 15747023 TI - [Application of autologous blood transfusion in oral and maxillofacial surgery]. AB - In recent years, autologous blood transfusion has been widely applied in many clinical fields, because of its advantages of avoiding transmission of blood contagious diseases, accidents of blood transfusion and immune reaction relating to homogenous blood transfusion. Autologous blood transfusion also has been an important approach to lessen operative blood loss during oral and maxillofacial surgery. This paper reviews the procedures, indications, contraindications and complications of autologous blood transfusion, as well as its application in oral and maxillofacial surgery. PMID- 15747024 TI - [Biomechanical study of rigid fixation of the maxillofacial fractures]. AB - Due to the ascending incidence rate of traffic trauma, more and more fractures in oral and maxillofacial region were encountered, which become more severe than before and damage the patient's quality of life and appearance. Biomechanical principles play an extremely important role in treatment of fractures,and study of biomechanical changes of fractured bone before and after rigid internal fixation will improve the effectiveness of therapy. The author summarized the biomechanical studies in the field of rigid internal fixation of fractures in oral and maxillofacial region. PMID- 15747025 TI - [Effect of Twin-block appliance in the treatment of Class II and division I malocclusion: a cephalometric study in 12 patients]. AB - PURPOSE: To quantitatively evaluate the skeletal and dentoalveolar changes induced by Twin-block appliance for treatment of Class II,division I malocclusions in growing children. METHODS: Twelve patients with Class II, division I malocclusions aged 10 to 12 years were investigated. The records were evaluated and compared with an age-matched sample of untreated Class II control subjects. The follow-up period was 12 months. Group t test was used for comparison. RESULTS: In the treatment group, the upper incisors retroclination of the upper incisors and the mandibular forward movement contributed more to the overjet reduction. The skeletal and dental effects changes molar correction were almost similar. The mandibular growth in the treatment group was 2.29 mm, greater than that in the control group. The posterior facial height and the anterior lower facial height increased by 2.28 mm and 2.58 mm respectively. On the other hand, unfavorable vertical changes were not observed. CONCLUSIONS: The treatment of Class II division I malocclusion with Twin-block appliance was effective in reducing overjet, correcting molar relationship and improving soft tissue profile. It can also enhance mandibular growth. PMID- 15747026 TI - [Treatment of primary trigeminal neuralgia with radiofrequency thermogangliocoagulation: Report of 152 consecutive cases]. AB - PURPOSE: To evaluate the clinical results of primary trigeminal neuralgia treated by RFT and investigate the methods to manage complications. METHODS: 152 cases of primary trigeminal neuralgia were collected from 2001 to 2004, a retrospective study was undertaken to analyze the clinical characters such as effective rate, ineffective rate and the rate of complications of patients treated with RFT. RESULTS: The average age of the 152 cases was 48.9 years old, the rate of male to female was 1:1.3. The effective rate of primary trigeminal neuralgia treated with RFT was 94.1%, the ineffective rate was 2.6%;another 3.3% cases gave up or altered to other methods, the rate of complications was 15.8%. CONCLUSIONS: RFT is an effective method to treat primary trigeminal neuralgia with few severe complications and low mortality rate, which can save the sense of touch partly. Recurrent cases can be retreated with RFT. PMID- 15747027 TI - The roles of endoscopic ultrasonography in the diagnosis of pancreatic tumors. AB - BACKGROUND: Over 20 years have passed since the development of endoscopic ultrasonography (EUS), and this technique is now widely used for the diagnosis of digestive and pancreaticobiliary diseases. EUS plays a number of roles in the clinical diagnosis of pancreatic tumors including detection of small tumors, differential diagnosis of tumors, and evaluation of tumor extent. METHODS: We evaluated the ability of EUS to detect pancreatic carcinoma in comparison with ultrasonography (US) and computed tomography (CT). RESULTS: The overall tumor detection rates were: US, 69%; CT, 79%; and EUS, 97%. With a tumor size smaller than 2 cm (TS1), the detection rate by EUS was as high as 100%, in contrast to 60% and 50% for US and CT, respectively, except for carcinoma in situ. However, EUS has some pitfalls. The EUS imaging technique is not always easy, EUS has limited penetration, and EUS alone cannot accurately differentiate malignancy from inflammation. CONCLUSIONS: The following points are the future themes of EUS. (1) It will be necessary to foster a large number of operators and to establish a standard EUS scanning technique. (2) Development of a new instrument with high resolution and deep penetration is anticipated. (3) Development of a new medium that can be infused into a vein and that allows diagnosis from B-mode images is much anticipated. PMID- 15747028 TI - Pancreatic tumors: role of imaging in the diagnosis, staging, and treatment. AB - Because most patients with pancreatic cancer present with biliary obstruction, percutaneous transhepatic cholangiopancreatography (PTC) or endoscopic retrograde cholangiopancreatography (ERCP) is often performed first to relieve obstruction. Fine needle biopsy (FNA) provides a tissue diagnosis, but is often nondiagnostic due to sampling error. Computed tomography (CT) is the workhorse of oncology, but is poor at defining the nature of pancreatic lesions. Small primary tumors are often not visualized. Fast magnetic resonance imaging (MRI) techniques allowing dynamic imaging after IV gadolinium and new contrast agents allow better characterization of the lesions for patients having contraindications for IV CT contrast agents. Magnetic resonance cholangiopancreatography (MRCP) allows noninvasive visualization of the biliary tree. Endoscopic ultrasonography (EUS) allows evaluation of the detailed regional anatomy with the possibility of FNA. 18F-Fluorodeoxyglucose (FDG) is the most common tracer used in positron emission tomography (PET), and most malignant tumors, including pancreatic carcinoma, have increased FDG uptake compared with normal cells. This functional imaging does not replace but is complementary to morphological imaging. FDG PET is particularly helpful: (1) for the diagnosis in patients with suspected pancreatic cancer in whom CT fails to identify a mass, or those in whom FNAs are nondiagnostic; (2) for staging by detecting CT-occult metastases; (3) for detecting recurrence; and (4) for monitoring therapy. Limitations include false-positive inflammatory processes and false-negative carcinoma in patients with diabetes and hyperglycemia, and islet cell tumors. PMID- 15747029 TI - Diagnostic laparoscopy in staging pancreatic carcinoma: developments during the past decade. PMID- 15747030 TI - Preoperative portal vein embolization: is it useful? AB - BACKGROUND/PURPOSE: Portal vein embolization (PVE) before hepatectomy is aimed to induce an atrophy of the embolized lobe to be resected, with a compensatory hypertrophy of the counterlobe to be preserved. METHODS: To answer the question "Is it useful?," we reviewed the clinical outcome in 161 patients undergoing major hepatectomy after PVE for various hepatobiliary tumors. RESULTS: All the patients tolerated PVE well, and hepatic functional data returned to the baseline levels within a week. The left liver volume increased by a median of 8% (range 2% 14%) after the right PVE. The 20 patients undergoing right hepatectomy for hepatocellular carcinoma had a mean indocyanine green retention rate at 15 min of 16% (SD 4%), and the 24 patients with liver metastases underwent right hepatectomy with additional left liver resection. Hepatectomy procedures comprised right or extended right hepatectomy (n=105), left or extended left hepatectomy (n=13), hepatopancreato duodenectomy (n=12), and less extensive hepatectomies (n=31). As a whole, the operative morbidity and mortality rates were 19% and 1.2%, respectively. Hepatopancreato duodenectomy carried no operative mortality. The cumulative 5-year survival rates were 44% in patients with hepatocellular carcinoma and 60% in patients with metastatic tumor. CONCLUSIONS: PVE is useful for performing extensive hepatectomy in patients with mild hepatic dysfunction, in those with bilobar tumors, or in those undergoing hepatopancreato duodenectomy. PMID- 15747031 TI - Arguments for a selective approach of preoperative portal vein embolization before major hepatic resection. AB - Preoperative PVE can induce hypertrophy of the future liver remnant volume resulting in a decrease of surgical risk after major hepatic resection. However, the number of patients with normal liver at risk is small and there is no arguments for inducing hypertrophy before standard right hepatectomy. Therefore, in patients with normal liver PVE is indicated in patients in whom very extended liver resection or associated major gastro-intestinal surgery is planned. In patients with chronic liver disease and in those with injured liver (chemotherapy, major steatosis, cholestasis), PVE is indicated before major liver resection. PMID- 15747032 TI - Small cell neuroendocrine carcinoma of the ampulla of Vater with foci of squamous differentiation: a case report. AB - A 54-year-old woman with obstructive jaundice was found to have a 4-cm ulcerated, elevated tumor, located at the papilla of Vater by endoscopy and radiographic investigation. Based on a clinical diagnosis of carcinoma of the ampulla of Vater, a pylorus-preserving pancreaticoduodenectomy was performed. The histologic appearance of the lesion was identical to extrapulmonary small cell carcinoma, with diffuse proliferation of small, spindle-shaped, atypical tumor cells with numerous mitoses. Neuroendocrine differentiation was demonstrated by immunoreactivity with neuron-specific enolase (NSE) and Leu-7, and by the presence of dense core granules ultrastructurally. The tumor was composed mainly of small cell neuroendocrine carcinoma, and partially of an area of squamous differentiation, showing transition from one to the other. Histopathologic investigation disclosed direct invasion deep to the lamina propria, and infiltration of the pancreatic parenchyma, duodenum, and bile duct. Lymph node metastases were present in the superior pancreaticoduodenal chain. The patient died of liver metastases 8 months later. As in previously reported cases of small cell neuroendocrine carcinoma of the ampullary region, the present case showed extremely aggressive clinical behavior with early metastases resulting in fatal outcome. PMID- 15747034 TI - [Prolapse surgery]. AB - Operative cystocele/rectocele management and prolapse surgery have become increasingly important domains of urogynecologic surgery. The risk of prolapse surgery in women lies at around 11% today and one-third of these are reoperations. There are currently three competing operative procedures: (1) the transvaginal approach with a vaginae fixatio sacrospinalis vaginalis or sacrotuberalis, (2) transabdominal pelvic sacrocolpopexy, and (3) transabdominal laparoscopic sacrocolpopexy.We compared efficacy, reoperation rates, and complication rates in abdominal sacrocolpopexy to those found with the transvaginal or laparoscopic surgical approaches, by reviewing the literature of the last 10 years, including 1995, in an online search. Analysis of the accumulated data made it clear, moreover, that randomized, prospective studies on the three different operative procedures are missing; these are mandatory, however, to enable exact and objective assessment of efficacy, long-term results, and complication rates. PMID- 15747033 TI - Molecular cloning and characterization of a novel adenylate kinase 3 gene from Clonorchis sinensis. AB - Adenylate kinase (AK) is a ubiquitous enzyme that contributes to the homeostasis of adenine nucleotides in living cells. AK catalyzes reversible high energy phosphoryl transfer reactions between ATP (or GTP) and AMP to generate ADP (or GDP). From a Clonorchis sinensis adult worm cDNA library, we isolated a cDNA clone encoding a novel AK3 isozyme. The 956 bp cDNA encodes a putative protein of 228 amino acids with a predicted molecular mass of 26.2 kDa. The recombinant CsAK3 protein produced in Escherichia coli can be refolded into a functional protein with AK3 activity. The optimum pH and temperature for the enzyme are 8.5 and 40 degrees C, respectively. The calculated activation energy is 56.04 kJ mol 1. The Km of the CsAK3 for AMP and GTP are 118 microM and 359 microM, respectively. CsAK3 is inhibited by Ap5A (>70% inhibition by 2.0 mM AP5A). Ap5A may be a potential lead compound acting on C. sinensis in which AK3 as a drug target. PMID- 15747035 TI - The influence of 4-week treatment with sertraline on the combined T3/TRH test in depressed patients. AB - In the present study, the influence of a 4-week treatment with sertraline on the regulation of hypothalamic-pituitary-thyroid (HPT) axis activity in depression was investigated, in particular the impact of sertraline on the thyroid receptor (TR)-mediated negative feedback control as measured by the combined T3/TRH test. In 20 drug-free patients (8 men, 12 women) suffering from a major depressive episode according to DSM-IV criteria the single TRH-stimulation test (administration of 200 microg TRH at 09:00h) was carried out followed by a combined T3/TRH test (pre-treatment with 40 microg 3,5,3'-triiodothyronine [T3] the night before; administration of 200 microg TRH at 09:00h the next day). After 4 weeks of treatment with sertraline at a standard dosage of 50 mg/day, both the single TRH test and the combined T3/TRH test were repeated in the depressed patients. Using repeated-measures ANOVA for statistical analysis, antidepressant therapy with sertraline did not have any significant impact on the TRH-induced TSH and prolactin stimulation (deltaTSH, deltaPRL) during the single TRH test nor during the combined T3/TRH test, neither in responders (n = 10) nor in non responders (n = 10). Moreover, the percentage suppression of TRH-induced TSH stimulation (deltaTSH) after pre-treatment with 40 microg T3 was comparable before (-61.07%) and after the 4-week treatment with sertraline (-58.92%). Apparently, the therapeutic efficacy of antidepressants such as sertraline is not related to the regulation of HPT axis activity in depressed patients. PMID- 15747036 TI - Atrial fibrillation: profit from cardiac pacing? AB - The impact of cardiac pacing on the prevention of atrial fibrillation is under scientific investigation. Several prospective randomised clinical trials have reported that atrial-based "physiologic" AAI(R)- or DDD(R)-pacing is associated with a lower incidence of paroxysmal and permanent atrial fibrillation than single-chamber ventricular pacing in patients with a conventional pacemaker indication. However, it is still uncertain whether atrial pacing itself has independent antiarrhythmic properties. In contrast, right ventricular pacing is considered to promote atrial fibrillation, even in preserved AV synchrony during dual-chamber pacing. The electrical secondary prevention of atrial fibrillation is mainly based on variations of the atrial pacing site and sophisticated preventive pacing algorithms incorporated in the pacemaker software. Dual-site right atrial and biatrial pacing were reported to exhibit modest to no benefit for the prevention of atrial fibrillation, whereas septal pacing and specific preventive pacing algorithms have been demonstrated to reduce the incidence of atrial fibrillation in a number of clinical trials. However, the role of septal pacing and preventive pacing algorithms still has to be clarified since, overall, study results have been inconsistent so far. One of the main goals of future investigations should be the identification of responder groups of preventive pacing concepts. In clinical practice, the efficacy of pacing algorithms and septal pacing has to be determined in the individual case. These options should be taken into account in patients with symptomatic bradycardia as the indication for cardiac pacing and, in addition, symptomatic atrial fibrillation. PMID- 15747037 TI - The first coronary angioplasties in Germany. PMID- 15747038 TI - Carotid artery stenting in clinical practice results from the Carotid Artery Stenting (CAS)-registry of the Arbeitsgemeinschaft Leitende Kardiologische Krankenhausarzte (ALKK). AB - BACKGROUND: Carotid artery stenting (CAS) for symptomatic and asymptomatic carotid stenosis seems to be on the doorstep of more widespread use. However, its feasibility and safety in clinical practice at a broad spectrum of hospitals needs to be determined. METHODS: We analyzed data of the prospective multi-centre Carotid Artery Stenting (CAS) Registry of the German Arbeitsgemeinschaft Leitende Kardiologische Krankenhausarzte (ALKK). RESULTS: From 7/1996 to 5/2004 1888 patients from 28 hospitals were included in the CAS Registry. The median hospital stay from CAS until hospital discharge was 3 days (quartiles: 2-6 days). Median patients age was 70 years (quartiles: 64-76 years) with 72.1% males. CAS for symptomatic stenosis was performed in 55% of cases. Patients treated with CAS suffered from coronary artery disease in 66.5%, had arterial hypertension in 91.7%, hyperlipidemia in 86.3% and 34.2% were diabetics. The intended CAS procedure was completed in 98.1% of cases. A stent was implanted in 97.3% of completed cases. The median intervention time was 43 min (quartiles: 30-60 min). During the hospital stay death rate was 0.3% (5/1888) and the rate of ipsilateral stroke 3.2% (58/1840). A contralateral ischemic event occurred in 1.4% (26/ 1840) of patients. The combined rate of all death or strokes was 3.8% (70/1841). Between 1996 and 2004 there was a steady increase in the use of protection devices during CAS (0% in 1996 and 97.9% in 2004; p for trend <0.0001). There was also an increase in the proportion of patients treated for asymptomatic stenoses (p for trend <0.0001). We observed a decrease of the combined endpoint of death or stroke from 6.3% in 1996 to 1.9% in 2004 (p for trend=0.021). CONCLUSION: The multi-centre ALKK CAS Registry data confirm the feasibility and shot-term safety of CAS even in daily clinical practice. There was a rapid penetration of the use of embolic protection devices, an increase in treatment of asymptomatic carotid stenoses and a decrease in acute complication rates from 1996 to 2004. PMID- 15747039 TI - Quantification of aortic valve area and left ventricular muscle mass in healthy subjects and patients with symptomatic aortic valve stenosis by MRI. AB - MRI allows visualization and planimetry of the aortic valve orifice and accurate determination of left ventricular muscle mass, which are important parameters in aortic stenosis. In contrast to invasive methods, MRI planimetry of the aortic valve area (AVA) is flow independent. AVA is usually indexed to body surface area. Left ventricular muscle mass is dependent on weight and height in healthy individuals. We studied AVA, left ventricular muscle mass (LMM) and ejection fraction (EF) in 100 healthy individuals and in patients with symptomatic aortic valve stenosis (AS). All were examined by MRI (1.5 Tesla Siemens Sonate) and the AVA was visualized in segmented 2D flash sequences and planimetry of the performed AVA was manually. The aortic valve area in healthy individuals was 3.9+/-0.7 cm(2), and the LMM was 99+/-27 g. In a correlation analysis, the strongest correlation of AVA was to height (r=0.75, p<0.001) and for LMM to weight (r=0.64, p<0.001). In a multiple regression analysis, the expected AVA for healthy subjects can be predicted using body height: AVA=-2.64+0.04 x(height in cm) -0.47 x w (w=0 for man, w=1 for female).In patients with aortic valve stenosis, AVA was 1.0+/-0.35 cm(2), in correlation to cath lab r=0.72, and LMM was 172+/-56 g. We compared the AS patients results with the data of the healthy subjects, where the reduction of the AVA was 28+/-10% of the expected normal value, while LMM was 42% higher in patients with AS. There was no correlation to height, weight or BSA in patients with AS. With cardiac MRI, planimetry of AVA for normal subjects and patients with AS offered a simple, fast and non-invasive method to quantify AVA. In addition LMM and EF could be determined. The strong correlation between height and AVA documented in normal subjects offered the opportunity to integrate this relation between expected valve area and definitive orifice in determining the disease of the aortic valve for the individual patient. With diagnostic MRI in patients with AS, invasive measurements of the systolic transvalvular gradient does not seem to be necessary. PMID- 15747040 TI - Self-management of oral anticoagulation in nonvalvular atrial fibrillation (SMAAF study). AB - Most patients with atrial fibrillation are at risk of suffering thromboembolic events. This risk can be reduced by twothirds by efficient anticoagulation. This prospective multi-center trial investigated whether the quality of treatment can be improved by self-management in patients with atrial fibrillations (SMAAF Study) compared to conventional patient management by the family doctor. METHODS: Two thousand patients suitable for self-management were to be randomized into the two arms of the study. In the period of investigation from December 1999 to July 2001, only 202 patients (64.3+/-9.2 years, 69.3% men) consented to participate. The study was discontinued prematurely since the number of patients was too low. As a consequence, the group comparison is confined to the evaluation of the INR values measured using the two-tailed t test. RESULTS: Of the 202 patients included, 101 were assigned to the self-management group (64.6+/-9.6 years, 71.4% men) and 101 (64.1+/-8.9 years, 61.4% men, n.s.) were assigned to the group managed by the family doctor. The total number of INR measurements was 2 865. This comprised 2 072 measurements in patients under self-management and 793 in the family doctor group. The values were within the target range significantly more frequently (p=0.0061) in patients under self-management (67.8%) as compared to the family doctor group (58.5%). There was a trend with regard to the time within target range, but the difference was not significant (178.8+/-126 days as compared to 155.9+/-118.4 days). In the self-management group, there were two severe hemorrhages, and there was one thromboembolic event in the family doctor group. CONCLUSION: Management of oral anticoagulation by INR self-management in patients with atrial fibrillation is not inferior to conventional care. PMID- 15747041 TI - Sirolimus-eluting stent treatment for isolated proximal left anterior descending artery stenoses. Results from the prospective multi-center German Cypher Registry. AB - BACKGROUND: Stenting of isolated proximal LAD stenoses is still a controversial issue since it is associated with higher target vessel revascularization (TVR) rate than both bypass surgery using the internal mammary artery, and stenting of other coronary artery territories. The sirolimus- eluting stent (SES) has been reported to significantly reduce restenosis rates in de novo coronary lesions. Therefore, we compared patients from the German Cypher Registry treated with SES for isolated proximal LAD lesions with those stented for isolated lesions in the proximal LCX or RCA. METHODS: A total of 349 patients treated with SES were analyzed. 249 patients were treated for proximal LAD stenosis, and 100 for proximal LCX/RCA stenoses. The combined clinical endpoint was MACCE (death of any cause, non-fatal MI and non-fatal stroke) and TVR at 6 months. RESULTS: In hospital events (death, MI and TVR) did not differ significantly between both groups (3.2% for the LAD group vs 2.0% for the LCX/RCA-group, p=0.73). The combined end point of death of any cause, non-fatal MI and non-fatal stroke at six months was 2.6% in the LAD group, and 2.2% in the LCX/RCA group (p=1.0). TVR occurred in 4.8% of the LAD group and in 6.5% of the LCX/RCA group at six months (p=0.58). The percentage of patients free from angina at daily activities was 80.6% in the LAD group, and 77.4% in the LCX/ RCA group (p=0.52). CONCLUSION: SES once implanted into isolated proximal LAD stenoses appears as effective as reported in other vessel territories. Accordingly, stenting of the proximal LAD using SES might prove a suitable alternative to surgery. PMID- 15747042 TI - Radiofrequency ablation of accessory pathways. Contemporary success rates and complications in 323 patients. AB - INTRODUCTION: 17 years ago the first radiofrequency catheter ablation of an accessory pathway (AP) was performed. The aim of this study was to describe the contemporary success rates and procedure related complication rates of radiofrequency (RF) ablation of accessory pathways (APs). In addition, the present study describes the anatomical distribution of APs according to the new nomenclature introduced by NASPE and ESC in 1999. METHODS: The analysis included all patients, who underwent RF ablation of an AP in the Heart Center Leipzig between January 2000 and December 2003. RESULTS: Over a 4 year period 336 APs were ablated in 323 patients. 201 APs (60%) presented with antegrade and retrograde conduction and showed preexcitation on ECG. For the remaining 135 APs (40%), only retrograde conduction over the AP was documented. According to the new nomenclature APs were classified as left-sided, right sided, septal and paraseptal APs. 188 APs (56%) were located on the left, 41 (12%) on the right, 64 (19%) in the paraseptal space and 31 APs (9%) presented with a septal or parahisian localization, respectively. Because of atypical course and/or characteristics 12 APs (4%) could not be classified. Ablation of all pathways were successful in 315 patients (98%). In 289 patients (89%) success was achieved within a single ablation session. The left-sided pathways had a re-intervention rate of 5%, which was significantly lower compared to the remaining localizations. The highest re-intervention rate was observed in the septal APs (23%). Complications were observed in less than 2% of all treated patients. CONCLUSIONS: 17 years after the first RF catheter ablation of an AP this therapy is established as a highly effective procedure. The success rate has improved to 98% and the complication rate has been minimized to less than 2%. The most frequent localization of APs is left posterior. Left sided APs also presented with the lowest re-intervention rate. The introduction of the new nomenclature in 1999 by NASPE and ESC has simplified the description of the exact anatomical localization of an AP. PMID- 15747046 TI - [20th report of performance data from heart catheterization laboratories in Germany. 2003 results of a collaborative survey by the Committee of Clinical Cardiology and the Interventional Cardiology (for ECS) and Angiology Study Groups of the German Society of Cardiology-Cardiovascular Research]. PMID- 15747043 TI - Splenic rupture complicating periinterventional glycoprotein IIb/IIIa antagonist therapy for myocardial infarction in polycythemia vera. AB - Polycythemia vera is a myeloproliferative disorder predisposing to thromboembolic and bleeding complications. We report the case of a patient with polyglobuly, leukocytosis, and thrombocytosis, who suffered from acute ST-segment elevation myocardial infarction due to thrombotic high-grade pre-stent stenosis two months after percutaneous coronary intervention for complex coronary one vessel disease. Following re-PTCA and stent implantation in conjunction with periinterventional GP IIb/IIIa antagonist treatment, the patient was initially symptom free for about two hours before rapidly developing signs of a hemorrhagic shock. An abdominal CT scan showed splenic rupture with massive intraabdominal hemorrhage as a consequence of secondary bleeding into multiple pre-existing splenic infarctions. The patient's condition stabilized after emergency splenectomy. Subsequent bone marrow biopsy revealed the presence of polycythemia vera. Post operatively, the patient was treated with the anti-platelet agents aspirin and clopidogrel to prevent subacute stent thrombosis. Additionally, cyto-reductive therapy with hydroxyurea was initiated because of a further increase in the platelet count. In patients with polycythemia vera, the indication for treatment with GP IIb/IIIa antagonists should be carefully weighed against the potentially serious bleeding complications. Should treatment be established, a risk stratification using abdominal sonography and bleeding time testing is recommended, while during treatment red blood count, platelet count, coagulation tests, and hemodynamic parameters should be closely monitored. PMID- 15747044 TI - [First diagnosis of Fallot tetralogy in a 74-year-old man]. AB - Tetralogy of Fallot is the most common cyanotic cardiac malformation in late childhood and adult, occurring in approximately 0.25 of 1000 live births. Most patients undergo early surgical correction. Therefore, the natural history of this disease has been evaluated in only a few cases. We report a complex case of a tetralogy of Fallot, who reached the age of 74 years without surgical or medical treatment and who was transferred to our clinic after syncope due to ventricular tachycardia. PMID- 15747048 TI - Current therapies in rabies. PMID- 15747049 TI - Nucleotide sequence analyses of genomic RNAs of Peanut stunt virus Mi, the type strain representative of a novel PSV subgroup from China. AB - The complete nucleotide sequence of Peanut stunt virus strain Mi (PSV-Mi) from China was determined and compared to other viruses of the genus Cucumovirus. The tripartite genome of PSV-Mi encoded five open reading frames (ORFs) typical of cucumoviruses. Distance analyses of four ORFs indicated that PSV-Mi differed sufficiently in nucleotide sequence from other PSV strains of subgroups I and II to warrant establishment of a third subgroup of PSV. PMID- 15747050 TI - Roles of the polypyrimidine tract and 3' noncoding region of hepatitis C virus RNA in the internal ribosome entry site-mediated translation. AB - Hepatitis C virus (HCV) genome contains a 3'noncoding region (3'NCR) consisting of a variable region, a polypyrimidine tract (polyU/UC) and the X region. To examine the roles of 3'NCR and polyU/UC tract in the internal ribosome entry site (IRES)-mediated translation process, a variety of 3'NCRs containing different lengths of polyU/UC tract were obtained from HCV infected patients and cloned respectively to the downstream of the firefly luciferase coding gene linked to HCV 5'NCR and 30 nucleotides of core gene (containing IRES element). The results of in vitro translation in rabbit reticulocyte lysate (RRL) and cell transfection assay in mammalian cells showed that the IRES-mediated translation efficiency could be enhanced by the full-length of 3'NCR of HCV RNA. However, contradictory results were observed when the role of polyU/UC tract in the IRES-mediated translation was studied. While the IRES-mediated translation efficiency was inhibited by the presence of polyU/UC tract in in vitro translation experiments, transfection of these expression cassettes into hepatic cell line showed that polyU/UC tract enhanced IRES-mediated translation efficiency in vivo. Cellular fraction complement experiments showed that cellular factors were required for the enhancement by the polyU/UC tract. Further antibody blocking assay and UV cross-linking assay suggested the correlation of IRES-mediated translation with host factors, including the La protein. The data above also indicated that the modulations of the IRES-mediated translation by the HCV 3'NCR and the polyU/UC tract were in a length-independent manner. PMID- 15747051 TI - Sequence analysis of genome segments S5 and S10 of Mal de Rio Cuarto virus (Fijivirus, Reoviridae). AB - Mal de Rio Cuarto virus (MRCV) was recently described as a new species of the genus Fijivirus, family Reoviridae. The nucleotide sequence of two MRCV genome segments was determined. MRCV S5 and S10 were predicted to encode proteins of 106.9 and 63.5 kDa respectively. The protein coded by MRCV S5 had 62.8% and 35.7% identity to fijiviruses RBSDV S5 and FDV S5 coded proteins, and contained a rarely reported type-1 C-terminal peroxisomal targeting signal. The protein coded by MRCV S10 had identity levels of 72.4% and 21.7% to the major outer capsid proteins of fijiviruses RBSDV S10 and NLRV S8. PMID- 15747053 TI - Subcellular localization and immunodetection of movement proteins of olive latent virus 1. AB - Polyclonal sera raised to Escherichia coli-expressed movement proteins encoded by ORF 3 (p8K) and ORF 4 (p6K) of olive latent virus 1, were used for their immunodetection in infected Nicotiana benthamiana plants. In subfractionated locally infected tissues 4 days post inoculation (d.p.i.) that were analysed by Western blot, p8K was found in the fast-sedimenting fractions P1 and P30 containing membranous material and/or cell organelles and, likely, the fibrous structures mentioned below, but not in the soluble protein-containing supernatant. No p6K could be detected in these extracts. In locally inoculated leaves p8K began to accumulate from 2 d.p.i onwards reaching its peak at 4 d.p.i. Intracellular immunogold labelling of cells from locally and systemically infected tissues localized p8K primarily in fibrous inclusions made up of thin filaments with a helical structure present in the cytoplasm of locally and systemically infected cells. In systemic infections a light and scattered labelling was observed in the cytoplasm and near the cell wall. The specific serum to p6K did not label the fibrous structures and failed to recognize its antigen in systemically and locally infected tissues except at 4 d.p.i., when scattered labelling was observed in the cytoplasm and near plasmodesmata. PMID- 15747052 TI - Pathogenicity of H5 influenza viruses for ducks. AB - Four H5N1 highly pathogenic avian influenza (HPAI) viruses and an avirulent reassortant H5N1 virus were tested for their pathogenicity in domestic ducks. A/chicken/Yamaguchi/7/04 (H5N1) (Ck/Yamaguchi/04) isolated from a dead bird during the HPAI outbreak in Japan and A/duck/Yokohama/aq-10/03 (H5N1) (Dk/Yokohama/03) isolated from duck meat at a quarantine inspection for importation from China replicated in multiple organs including the brain of ducks. The ducks infected with Ck/Yamaguchi/04 did not show any clinical signs, while those infected with Dk/Yokohama/03 showed neurological signs. The ducks infected either with A/Hong Kong/483/97 (H5N1) or A/tern/South Africa/61 (H5N3), or with an avirulent H5N1 reassortant, did not show any clinical signs. Virus specific antibodies were detected in the sera of the ducks infected with each of the five strains tested, indicating that all of the viral strains infected and replicated in the birds. Dk/Yokohama/03 grew in multiple organs more rapidly than did Ck/Yamaguchi/04. Considerable titers of virus were detected in the brain of the ducks infected with Dk/Yokohama/03 and these birds showed neurological signs. The present results demonstrate that the pathogenicity of influenza viruses for ducks does not correlate with that for chickens and that replication of the virus in the brain is critical for ducks to show neurological signs. PMID- 15747054 TI - Bone metabolism in male patients with type 2 diabetes. AB - Few reports are available on bone turnover in type 2 diabetes. Impaired bone turnover in type 2 diabetes appears to result from decreased bone formation. Studies also suggest that poor glycaemic control in type 2 diabetes may contribute to osteopaenia. The aim of this study was to investigate biochemical markers of bone turnover in males with poorly controlled type 2 diabetes and look for correlations with glycaemic control and gonadal and hypophyseal hormonal axis. Consecutive male patients with poorly controlled type 2 diabetes and attending the internal medicine department during a period of 6 months were enrolled. The patients were receiving oral hypoglycaemic agents (metformin or sulphonylureas or both). None of the patients had any evidence of macroangiopathy, nephropathy or neuropathy. Only two patients had proliferative retinopathy. Serum osteocalcin, crosslaps (C-telopeptide, CTx), parathyroid hormone (PTH), testosterone, oestrogen, prolactin, follicle-stimulating hormone (FSH) and luteinising hormone (LH) were measured in 35 patients and 35 controls. The mean age of the study population was 53.7 (10.3) years (range: 50.2-57.3) and the mean disease duration was 8.6 (6.0) years (range: 6.5-10.7). No differences between patients and controls were observed in serum calcium, phosphorus, creatinine, albumin, PTH, CTx, oestrogen, testosterone, LH, FSH, prolactin and urinary calcium. Patients had lower serum levels of osteocalcin than controls with a significant statistical difference [15.3 (4.1) vs 18.3 (5.3), p=0.012]. There was a negative significant statistical correlation between CTx levels and HbA1c (r=-0.41, p< 0.05). Our study suggested that bone formation is altered in type 2 diabetes and that bone turnover is affected by glycaemic control status. PMID- 15747055 TI - Observations on parturition and allomothering in wild capped langur (Trachypithecus pileatus). AB - A birth during the day by a capped langur (Trachypithecus pileatus Blyth, 1843) was recorded at Pakhui Wildlife Sanctuary, Arunachal Pradesh, India. The birth took 43 min. Allomothering was observed 3 h after the birth. An average of 9% of daily active time was shared by four allomothers (three adults, one subadult) during the first 15 days of the infant's life. Total time allomothering was proportional to the age of the allomothers (241 min for oldest; 214 min for youngest). PMID- 15747056 TI - Cis-trans isomerase gene in psychrophilic Pseudomonas syringae is constitutively expressed during growth and under conditions of temperature and solvent stress. AB - In a recent study, we established that psychrophilic Pseudomonas syringae (Lz4W) requires trans-monounsaturated fatty acid for growth at higher temperatures (Kiran et al. in Extremophiles, 2004). It was also demonstrated that the cti gene was highly conserved and exhibited high sequence identity with cti of other Pseudomonas spp. (Kiran et al. in Extremophiles, 2004). Therefore it would be interesting to understand the expression of the cti gene so as to unravel the molecular basis of adaptation of microorganisms to high temperature. In the present study, the expression of cti was monitored by RT-PCR analysis during different growth stages and under conditions of high temperature and solvent stress in P. syringae. Results indicated that the cti gene is constitutively expressed during different stages of growth and the transcript level is unaltered even under conditions of temperature and solvent stress implying that the observed increase in trans-monounsaturated fatty acids (Kiran et al. in Extremophiles, 2004) is not under transcriptional control. A putative promoter present in the intergenic region of the metH and cti gene has also been characterized. The translation start site ATG, the Shine-Dalgarno sequence AGGA and the transcription start site "C" were also identified. These results provide evidence for the first time that the cti gene is constitutively expressed under normal conditions of growth and under conditions of temperature and solvent stress thus implying that the Cti enzyme is post-transcriptionally regulated. PMID- 15747057 TI - Exploring oligodendrocyte guidance: 'to boldly go where no cell has gone before'. AB - Oligodendrocytes, the myelinating cells of the central nervous system (CNS), originate early in the formation of the brain in specific foci, and migrate throughout the parenchyma. The instructional cues guiding the migration of these progenitor cells must be encoded into their developing environment. Soluble factors as well as membrane-bound cues most likely synergize to create a complex thoroughfare needed to sculpt and organize the brain into a functional organ with white and gray matter. Classically, the focus of many guidance related studies in the CNS has been limited to neuron physiology. However, It is becoming increasingly clear that their lifelong partners, oligodendrocytes, express both ligands and receptors able to both present and respond to these classical cues. In this short review, some recent findings in the Semaphorin and Eph fields will be presented with respect to oligodendrocyte expression and function. PMID- 15747058 TI - The potential role of endogenous bacteriophages in controlling invading pathogens. AB - Bacteriophages (phages) are omnipresent in our environment, and recent studies highlight their potential impact on the microbial world. Phages can also be present in mammalian organisms, including man (intestines, oral cavity, urine, sputum and serum). Data are available which suggest that those endogenous phages could play an important role in eliminating bacteria and regulating the body ecosystem. Furthermore, our most recent findings suggest that phages can exert immunosuppressive action in the gut, helping control local inflammatory and autoimmune reactions, and demonstrate anticancer activity. We hypothesize that phages could act in concert with the immune system in immunosurveillance against bacteria, viruses and cancer. PMID- 15747059 TI - Catalysis in abiotic structured media: an approach to selective synthesis of biopolymers. AB - Micro- and nanoenvironments formed by amphiphile self-assembled structures, water ice lattices and minerals have well-defined, repeating, chemical and physical properties that can be used for selective synthesis of biopolymers, such as RNAs and proteins. The advances made in the development of polymerization supported by these micro- and nanosystems are reviewed here. In particular, it is shown that these systems promote non-enzymatic biopolymerization, yielding long polymers whose sequence composition is determined by the interactions between monomers and the supporting environment. When used to compartmentalize enzymatic biopolymerization, micro- and nanostructures allow the implementation of molecular selection and evolution schemes, which are difficult in homogeneous medium, yielding very active molecules. Thus, micro- and nanoenvironment approaches to the synthesis and selection of biopolymers could be developed into a new biotechnological tool for the production of biopolymers with novel functions. PMID- 15747060 TI - Study of molecular events in cells by fluorescence correlation spectroscopy. AB - To understand processes in a living cell, sophisticated and creative approaches are required that can be used for gathering quantitative information about large number of components interacting across temporal and spatial scales without major disruption of the integral network of processes. A physical method of analysis that can meet these requirements is fluorescence correlation spectroscopy (FCS), which is an ultrasensitive and non-invasive detection method capable of single molecule and real-time resolution. Since its introduction about 3 decades ago, this until recently emerging technology has reached maturity. As commercially built equipment is now available, FCS is extensively applied for extracting biological information from living cells unattainable by other methods, and new biological concepts are formulated based on findings by FCS. In this review, we focus on examples in the field of molecular cellular biology. The versatility of the technique in this field is illustrated in studies of single-molecule dynamics and conformational flexibility of proteins, and the relevance of conformational flexibility for biological functions regarding the multispecificity of antibodies, modulation of activity of C5a receptors in clathrin-mediated endocytosis and multiplicity of functional responses mediated by the p53 tumor suppressor protein; quantitative characterization of physicochemical properties of the cellular interior; protein trafficking; and ligand-receptor interactions. FCS can also be used to study cell-to-cell communication, here exemplified by clustering of apoptotic cells via bystander killing by hydrogen peroxide. PMID- 15747062 TI - Legionella pneumophila down-regulates MHC class I expression of human monocytic host cells and thereby inhibits T cell activation. AB - Legionella (L.) pneumophila, the causative agent of Legionnaires' disease, is an intracellular pathogen of alveolar macrophages that resides in a compartment displaying features of endoplasmatic reticulum (ER). In this study, we show that intracellular multiplication of L. pneumophila results in a remarkable decrease in MHC class I expression by the infected monocytes. During intracellular multiplication, L. pneumophila absorbs ER-resident chaperons such as calnexin and BiP, molecules that are required for the correct formation of the MHC class I complex. Due to reduced MHC class I expression, stimulation of allogeneic blood mononuclear cells was severely inhibited by infected host cells but cytotoxicity of autologous natural killer cells against Legionella-infected monocytes was not enhanced. Thus, reduced expression of MHC class I in infected monocytes may resemble a new immune escape mechanism induced by L. pneumophila. PMID- 15747063 TI - Anti-glioma action of aloe emodin: the role of ERK inhibition. AB - The effect of aloe emodin (AE), a herbal anthraquinone derivative, on the rat C6 glioma cell line was investigated. In addition to cell cycle block and caspasedependent apoptosis, AE led to the formation of intracytoplasmic acidic vesicles indicative for autophagic cell death. Moreover, differentiation of surviving cells toward the astrocytic lineage was confirmed by typical morphological changes and increased expression of glial fibrillary acidic protein (GFAP). AE did not affect the activation of mitogen-activated protein kinase p38, Jun-N-terminal kinase, or transcription factor NF-kappaB, but markedly inhibited the activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) in C6 cells. A selective inhibitor of ERK activation, PD98059, mimicked the effects of AE on glioma cell morphology and GFAP expression, but failed to induce either apoptosis or autophagy. Taken together, these results indicate that the anti glioma action of AE involves ERK-independent induction of both apoptosis and autophagy, as well as ERK inhibition-mediated differentiation of glioma cells. PMID- 15747065 TI - A second protein kinase CK1-mediated step negatively regulates Wnt signalling by disrupting the lymphocyte enhancer factor-1/beta-catenin complex. AB - Deregulated activation of the canonical Wnt signalling pathway leads to stabilization of beta-catenin and is critically involved in carcinogenesis by an inappropriate induction of lymphocyte enhancer factor (LEF-1)/beta-catenin dependent transcription of Wnt target genes. Phosphorylation of the pathway components beta-catenin, Dishevelled, Axin and APC (adenomatous polyposis coli) by glycogen synthase kinase-3beta, CK1 and CK2 is of central importance in the regulation of the beta-catenin destruction complex. Here, we identify CK1 and CK2 as major kinases that directly bind to and phosphorylate LEF-1 inducing distinct, kinase-specific changes in the LEF-1/DNA complex. Moreover, CK1-dependent phosphorylation in contrast to CK2 disrupts the association of beta-catenin and LEF-1 but does not impair DNA binding of LEF-1. Sequential phosphorylation assays revealed that for efficient disruption of the LEF-1/beta-catenin complex, beta catenin also has to be phosphorylated. Consistent with these observations, CK1 dependent phosphorylation inhibits, whereas CK2 activates LEF-1/beta-catenin transcriptional activity in reporter gene assays. These data are in line with a negative regulatory function of CK1 in the Wnt signalling pathway, where CK1 in addition to the beta-catenin destruction complex at a second level acts as a negative regulator of the LEF-1/beta-catenin transcription complex, thereby protecting cells from development of cancer. PMID- 15747064 TI - Alpha-crystallin: an ATP-independent complete molecular chaperone toward sorbitol dehydrogenase. AB - alpha-Crystallin, the major component of the vertebrate lens, is known to interact with proteins undergoing denaturation and to protect them from aggregation phenomena. Bovine lens sorbitol dehydrogenase (SDH) was previously shown to be completely protected by alpha-crystallin from thermally induced aggregation and inactivation. Here we report that alpha-crystallin, in the presence of the SDH pyridine cofactor NAD(H), can exert a remarkable chaperone action by favoring the recovery of the enzyme activity from chemically denaturated SDH up to 77%. Indeed, even in the absence of the cofactor, alpha crystallin present at a ratio with SDH of 20:1 (w:w) allows a recovery of 35% of the enzyme activity. The effect of ATP in enhancing alpha-crystallin-promoted SDH renaturation appears to be both nonspecific and to not involve hydrolysis phenomena, thus confirming that the chaperone action of alpha-crystallin is not dependent on ATP as energy donor. PMID- 15747067 TI - Management and outcome of pouch-vaginal fistulas following restorative proctocolectomy. AB - PURPOSE: The aim of this study was to assess the short-term and long-term outcomes of surgical repair of patients with pouch-vaginal fistulas after restorative proctocolectomy. METHODS: A descriptive study was undertaken of all patients developing pouch-vaginal fistulas following restorative proctocolectomy between 1978 and 2003 in a single tertiary referral institution. Kaplan-Meier survival analysis was used to evaluate the time to first pouch-vaginal fistula recurrence and pouch-vaginal fistula-free survival at last follow-up. RESULTS: Sixty-eight patients (mean age, 32.2 years; standard deviation, 10.7) were identified with a median follow-up of 5.5 (range, 0.2-25.5) years. The origin of the pouch-vaginal fistulas was the pouch-anal anastomosis in 52 (76.5 percent) patients, pouch body/top in 9 (13.2 percent), or cryptoglandular or other source in 7 (10.3 percent). Associated early complications in patients with pouch vaginal fistulas included pelvic sepsis in 20 (29 percent) patients, anastomotic separation in 6 (24 percent), anastomotic stricture in 16 (24 percent), small bowel obstruction in 17 (25 percent), hemorrhage in 2 (3 percent), or pouchitis in 12 (18 percent). Surgery was undertaken in 59 (87 percent) patients with 14 (20.6 percent) of them undergoing pouch excision/diversion or seton drainage. Forty-five (66 percent) patients underwent primary repair. First recurrence of pouch-vaginal fistula occurred in 27 of 45 (60 percent) patients with a median pouch-vaginal fistula-free interval of 1.6 years (95 percent confidence interval, 0.6-2.7). Fourteen (51.9 percent) patients with recurrent pouch-vaginal fistulas healed following one or more repeat procedures. The diagnosis of Crohn's disease was made in eight (12 percent) patients, with pouch-vaginal fistulas persisting or recurring in all patients with Crohn's disease within five years of the primary treatment. Median pouch-vaginal fistula-free survival was 1.4 years for patients with Crohn's disease and 8.1 years for patients with ulcerative colitis or familial adenomatous polyposis. The pouch-vaginal fistula-free survival improved with repeated local or abdominal repairs for patients with ulcerative colitis. The overall pouch failure rate for patients with pouch-vaginal fistulas was 35 percent (median pouch survival, 4.2 years). CONCLUSIONS: Pouch-vaginal fistulas can persist and recur indefinitely, even after repeated repairs. Repair in those patients with Crohn's disease uniformly failed within five years from primary repair. Patients with recurrent pouch-vaginal fistulas and ulcerative colitis should be offered salvage surgery because successful closure following initial failure occurs in approximately 50 percent. PMID- 15747068 TI - Topical 5-fluorouracil in the management of extensive anal Bowen's disease: a preferred approach. AB - PURPOSE: An alternative approach to anal Bowen's disease was investigated. The use of topical 5 percent 5-fluorouracil for large lesions and surgical excision of small lesions were evaluated. METHODS: A prospective study was undertaken for anal Bowen's disease in 11 patients over a six-year period. Before therapy all patients underwent anal mapping biopsy and colonoscopy. For one-half circumferential disease or greater, patients underwent topical 5 percent 5 fluorouacil therapy for 16 weeks. For smaller involvement, wide surgical excision was performed. All patients underwent anal mapping biopsy one year after completion of therapy. RESULTS: Of 11 patients, 8 (5 male) received 16 weeks of topical 5 percent 5-fluorouacil therapy. Three patients (3 female) underwent surgical excision for localized disease. All but one patient, who was HIV positive, were free of Bowen's disease one year after completion of therapy. One patient underwent total excision of a residual microinvasive squamous carcinoma after circumferential Bowen's disease had resolved. One patient received eight additional weeks of topical 5-fluorouacil therapy for incomplete resolution. All patients were followed yearly, with a mean follow-up of 39 months and a range of 12 to 74 months. There have been no recurrences. There were no long-term side effects or morbidity from topical 5-fluorouacil or local excision. All colonoscopies were normal. CONCLUSION: Topical 5 percent 5-fluorouacil therapy is a safe and effective method to treat anal Bowen's disease. Wide local excision is appropriate for smaller, isolated areas of disease. Anal Bowen's disease was not associated with colonic or other neoplasms. PMID- 15747069 TI - Immediate radical resection after local excision of rectal cancer: an oncologic compromise? AB - PURPOSE: Local excision for early-staged rectal cancers is controversial. Preoperative understaging is not uncommon and radical resection after local resection may be needed for a curative treatment. The aim of this study was to determine the frequency and outcome of radical resection (within 30 days) after local excision for rectal adenocarcinoma. METHODS: All locally excised rectal cancers (curative intent) that required radical surgery within 30 days were reviewed (1980-2000). T2-3N0-1 stage cancers were each matched to three primary radical surgery controls for stage, age (+/- 5 years), gender, date (+/- 1 years), and type (abdominoperineal resection or low anterior resection) of operation. T1N0-1 cancers were compared with stage-matched rectal cancers treated by either primary radical surgery (n = 78) or local excision alone (n = 77). RESULTS: Fifty-two locally excised rectal adenocarcinomas (29 transanal and 23 polypectomies) were followed by radical surgery (24 abdominoperineal resection and 28 low anterior resection) within 7 (range, 1-29) days. Radical surgery was performed because of a cancerous polyp (n = 42), positive margins (5), lymphovascular invasion (3), and T3-staged cancer (2). Twelve of 52 cancers (23 percent) were found to have nodal involvement and 15 of 52 (29 percent) showed residual cancer in the resected specimen. The T2-3N0-1 stage controls were well matched. No significant difference in tumor location, size, adjuvant therapy, or length of follow-up was noted. Local and distant recurrence occurred in 2 of 4 T2 3N1 tumors and in 2 of 11 T2-3N0 cancers and were comparable to the matched controls, as was survival, with the exception of shorter survival in T3N1 cases, but numbers were too small for a definitive conclusion. Length of follow-up was not different. For T1 cancers, the controls were also comparable regarding patient and tumor demographics and adjuvant therapy. Nodal involvement was 21 percent in T1 study cases and 15 percent in T1 primary radical-surgery controls, with a trend toward location in the lower third of the rectum in both groups (58 percent and 50 percent, respectively). Local recurrence rates were 3 percent in the study group, 5 percent for patients undergoing primary radical surgery, and 8 percent for local excision alone. Distant metastasis (11 percent, 12 percent, and 13 percent, respectively) and overall five-year survival were also not significantly different (78 percent, 89 percent, and 73 percent, respectively). CONCLUSIONS: Nodal involvement in attempted locally excised rectal cancers is not uncommon. Local excision of rectal tumors followed by radical surgery within 30 days in cancer patients does not compromise outcome compared with primary radical surgery. Even after radical surgery for superficial T1 rectal cancers, recurrence rates are not insignificant. Future improvements in preoperative staging may be helpful in selecting tumors for local excision only. PMID- 15747070 TI - Treatment of chronic anal fissure by application of L-arginine gel: a phase II study in 15 patients. AB - PURPOSE: Local application of exogenous nitric oxide donors, such as isosorbide dinitrate and glyceryl trinitrate, promotes fissure healing by reducing anal resting pressure and improving anodermal blood flow. The major drawback of these nitric oxide donors is headache. The overall incidence of this side effect is approximately 40 percent. Recently we have shown in healthy volunteers that L arginine, being an intrinsic precursor of nitric oxide, reduces anal resting pressure without headache as a side effect. The aim of the present study was to evaluate the effect of L-arginine on anal resting pressure, anodermal blood flow, and fissure healing in patients with chronic anal fissure. METHODS: Fifteen patients with a chronic anal fissure were included in the present study. Before entering the study 10 patients were unsuccessfully treated by local application of isosorbide dinitrate. Six of these patients experienced severe headache during treatment with isosorbide dinitrate. All patients were treated for at least 12 weeks by local application of a gel containing L-arginine 400 mg/ml five times a day. In patients with a persistent fissure, treatment was continued until 18 weeks. Anal manometry and laser Doppler flowmetry of the anoderm were performed before treatment, 20 minutes after local application of the first dose, and after 12 weeks of treatment. A visual analog scale was used to assess fissure-related pain and headache. RESULTS: One patient dropped out after one day of treatment, and one was excluded because of violation of the study protocol. After 12 weeks of treatment complete fissure healing was observed in 3 of 13 (23 percent) patients, and after 18 weeks the healing rate was 8 of 13 (62 percent) patients. None of the 13 patients experienced typical nitric oxide-induced headache. The pressure recordings showed a significant reduction of maximum anal resting pressure (mean +/- SD): pretreatment 89 +/- 17 mmHg; 20 minutes after application of the first dose 67 +/- 17 mmHg; 12 weeks after treatment 74 +/- 14 mmHg (P < 0.005). Recordings of anodermal blood flow showed a significant increase in flow: pretreatment 0.36 +/- 0.25 volts; 20 minutes after application of the first dose 0.59 +/- 0.27; 12 weeks after treatment 0.64 +/- 0.33 (P < 0.005). CONCLUSIONS: Local application of L-arginine promotes fissure healing without headache as a side effect, and L-arginine is effective even in patients not responding to isosorbide dinitrate treatment. PMID- 15747071 TI - Long-term outcome of mesocolic and pelvic diverticular abscesses of the left colon: a prospective study of 73 cases. AB - PURPOSE: The aim of of this study was to evaluate prospectively the long-term outcome of mesocolic and pelvic diverticular abscesses of the left colon. METHODS: Between October 1986 and October 1997, a total of 465 patients urgently admitted to our hospital with a suspected diagnosis of acute left-sided colonic diverticulitis had a CT scan. Of 76 patients (17 percent) who had an associated mesocolic or pelvic abscess, 3 were lost to follow-up. The remaining 73 patients (45 with a mesocolic abscess and 28 with a pelvic abscess) were followed for a median of 43 months. RESULTS: of the 45 patients with a mesocolic abscess, 7 (15 percent) required surgery during their first hospitalization versus 11 (39 percent) of the 28 patients with a pelvic abscess (P = 0.04). At the end of follow-up, 22 (58 percent) of the 38 patients with a mesocolic abscess who had successful conservative treatment during their first hospitalization did not need surgical treatment vs. 8 (47 percent) of the 17 who had a pelvic abscess. Altogether, 51 percent of the patients with a mesocolic abscess had surgical treatment versus 71 percent of those with a pelvic abscess (P = 0.09). CONCLUSIONS: Considering the poor outcome of pelvic abscess associated with acute left-sided colonic diverticulitis, percutaneous drainage followed by secondary colectomy seems justified. Mesocolic abscess by itself is not an absolute indication for colectomy. PMID- 15747072 TI - Surveillance-detected hepatic metastases from colorectal cancer had a survival advantage in seven-year follow-up. AB - PURPOSE: Colorectal cancer is a common cause of cancer-related death. The liver is the most common site of distant metastases and the most amenable to potentially curative surgery. The aim of this study was to determine whether hepatic metastases detected by surveillance following colonic resection were associated with higher resectability rates and to determine whether there was any impact on survival rates. METHODS: A retrospective study of 211 patients who presented to the liver unit between February 1990 and July 1996 with hepatic metastases following colonic resection for adenocarcinoma was performed. Patients were divided into two groups: Group A (n = 154), hepatic metastases diagnosed by carcinoembryonic antigen or by radiology; and Group B (n = 57), patients with symptomatic presentation. RESULTS: Potentially curative operations were possible in 51.3 percent (79/154) of Group A patients and 28.1 percent (16/57) of Group B patients (P = 0.0043, chi-squared test). In Groups A and B, 24 percent (37/154) and 43.9 percent (25/57) of patients, respectively, were inoperable. The three year and five-year survival rates after detection of liver metastases were 26.8 percent (41/153) in Group A and 12.5 percent (7/56) in Group B, and 5.9 percent (9/153) in Group A and 8.9 percent (5/56) in Group B, respectively. Log-rank analysis resulted in P = 0.05, Breslow test in P = 0.01. CONCLUSION: Our study shows that patients with hepatic metastases from colorectal cancer detected by follow-up were significantly more likely to have a potentially curative operation. Our medium-term survival data show a statistically significant survival benefit in patients with surveillance-detected metastases. PMID- 15747061 TI - G-protein signaling: back to the future. AB - Heterotrimeric G-proteins are intracellular partners of G-protein-coupled receptors (GPCRs). GPCRs act on inactive Galpha.GDP/Gbetagamma heterotrimers to promote GDP release and GTP binding, resulting in liberation of Galpha from Gbetagamma. Galpha.GTP and Gbetagamma target effectors including adenylyl cyclases, phospholipases and ion channels. Signaling is terminated by intrinsic GTPase activity of Galpha and heterotrimer reformation - a cycle accelerated by 'regulators of G-protein signaling' (RGS proteins). Recent studies have identified several unconventional G-protein signaling pathways that diverge from this standard model. Whereas phospholipase C (PLC) beta is activated by Galpha(q) and Gbetagamma, novel PLC isoforms are regulated by both heterotrimeric and Ras superfamily G-proteins. An Arabidopsis protein has been discovered containing both GPCR and RGS domains within the same protein. Most surprisingly, a receptor independent Galpha nucleotide cycle that regulates cell division has been delineated in both Caenorhabditis elegans and Drosophila melanogaster. Here, we revisit classical heterotrimeric G-protein signaling and explore these new, non canonical G-protein signaling pathways. PMID- 15747073 TI - How accurate is magnetic resonance imaging in restaging rectal cancer in patients receiving preoperative combined chemoradiotherapy? AB - PURPOSE: Preoperative combined chemoradiotherapy is currently the main neoadjuvant therapy used to treat locally advanced middle and low rectal adenocarcinoma. A restaging work-up with magnetic resonance imaging was hoped to provide information about the effects related to combined chemoradiotherapy. The goal was to evaluate the correlation between pathologically verified tumor stages and clinical stages predicted by magnetic resonance imaging after combined chemoradiotherapy. METHODS: Between August 2000 and June 2003, 50 patients with biopsy-proven middle and lower rectal adenocarcinoma, with initial stage T3-T4 or N+, M0, were recruited in this series. Pelvic magnetic resonance imaging was used to stage the tumor before and after combined chemoradiotherapy. A protocol of the standard external radiation dose and oral combined uracil and 5-fluorouracil plus leucovorin was used. The results of magnetic resonance imaging restaging after combined chemoradiotherapy were correlated with the pathologic staging. RESULTS: The overall predictive accuracy in T stage was 52 percent, whereas overstaging and understaging occurred in 38 percent and 10 percent of patients, respectively. Most of the inaccurate T staging was a result of the overstaging of superficial tumors (T0-T2). In N stage, accurate staging was noted in 68 percent of all patients, whereas 24 percent were overstaged and 8 percent were understaged. CONCLUSION: In restaging irradiated tumors, magnetic resonance imaging had the accuracy of 52 percent in T stage and 68 percent in N stage. Poor agreement between post-combined chemoradiotherapy magnetic resonance imaging and pathologic staging was observed in both T (k = 0.017) and N (k = 0.031) stages. Most of the inaccuracy in both T and N stages was caused by overstaging. The problem with magnetic resonance imaging was believed to be that it could not completely differentiate fibrosis from viable residual tumors. PMID- 15747074 TI - Rectoanal intussusception: presentation of the disorder and late results of resection rectopexy. AB - BACKGROUND: Rectoanal intussusception may cause symptoms of obstructed defecation, and functional results of prosthesis rectopexy are usually not satisfactory. The aim of this study was to assess several parameters of the disorder and to evaluate the outcome of resection rectopexy. METHODS: During a 10 year period, 27 female patients with symptomatic large rectoanal intussusception had resection rectopexy (23 laparoscopy; 4 laparotomy). Conservative treatment, including biofeedback treatment in 22 patients, had failed in all cases. Preoperative and postoperative evaluation included clinical assessment, anorectal manometry, evacuation defecography, and colon transit studies. Follow-up ranged between one and five years. RESULTS: Length of intussusception was 2 to 4.9 cm and was significantly related to pelvic floor descent (P = 0.003) and inversely related to resting anal pressures (P < 0.001). Eleven patients had undergone a previous hysterectomy, 9 had enterocele-sigmoidocele, 7 had incontinence of varying severity, and 8 had a solitary rectal ulcer. Colon transit was abnormal in all but five cases. Immediate functional results were bad in two-thirds of the cases; tenesmus, urge to defecate, and frequent stools were the main complaints. By the time these symptoms had subsided, and one year after surgery, all but two patients were satisfied with the outcome. Intussusception was reduced in all cases, anal sphincter tone recovered (P = 0.002), perineal descent decreased (P < 0.001), and colonic transit was accelerated (P < 0.001). Patients available at five-year follow-up had no or only minor defecatory problems. CONCLUSION: Resection rectopexy improves symptoms of obstructed defecation attributed to large rectoanal intussusception. PMID- 15747075 TI - The strictured anastomosis: successful treatment by corticosteroid injections- report of three cases and review of the literature. AB - Clinically significant anastomotic strictures usually only occur with very low colorectal anastomoses below the level of the peritoneal reflection. The reported rate averages 8 percent and has been attributed to tissue ischemia, localized sepsis, anastomotic leak, proximal fecal diversion, radiation injury, inflammatory bowel disease, and recurrent rectal cancer. Most patients will have symptoms of obstipation, frequent small bowel movements, and bloating. Symptomatic strictures are often approached by dilation (balloon or Hegar) or less often repeat resection. Many of these patients have anastomoses that are too low to consider repeat resection. Strictureplasty with linear stapling devices, stricture resection by use of the circular stapling device, and repeat dilations have all been described. Steroid injections into the stricture have been described in strictured esophagogastric anastomoses but have not been commonly used for strictured coloproctostomies. We describe three cases of coloanal stricture following resections that were complicated by postoperative pelvic abcesses, anastomatic leaks, and pelvic fibrosis. Two cases had undergone low coloanal anastomosis that was protected by a loop ileostomy and developed as significant stricture in the early postoperative period. The third case was managed without a protective loop ileostomy. These were initially managed by repeated dilation of the anastomosis. Each episode was followed by rapid recurrence of the stricture. All patients underwent subsequent dilation with injection of 40 mg of triamcinolone acetate (divided dose in four quadrants) into the stricture and subsequent complete resolution of the stricture. Those patients with loop ileostomies had them taken down and all have been followed for up to 12 months without clinical or endoscopic evidence of recurrent stricture. PMID- 15747076 TI - Prevalence and morphology of pouch and ileal adenomas in familial adenomatous polyposis. AB - PURPOSE: In familial adenomatous polyposis, the long-term risk of pouch polyposis and potential for pouch cancer are unknown. Our aim was to evaluate prospectively the prevalence, nature, and etiology of pouch ileal adenomas with that of nonpouch ileal adenomas in familial adenomatous polyposis. METHODS: Sixty patients with familial adenomatous polyposis pouch, 47 familial adenomatous polyposis patients with ileorectal anastomosis, and 20 younger patients with familial adenomatous polyposis who had prophylactic colectomy were examined with videoendoscopy. RESULTS: Adenomatous polyps were found in the pouches of 34 patients (57 percent). A total of 362 polyps were identified (range, 0-50 per patient). A logistic regression model confirmed that there was a significant association between the increasing age of the patient and the presence of pouch adenomas (P < 0.02) and the length of follow-up since pouch surgery (P < 0.05). There was no apparent relationship between the development of pouch adenomas and the severity of either colonic or duodenal polyposis and there were no clear genotype or phenotype correlations. Most polyps were tubular adenomas with mild dysplasia, but 11 patients had more advanced histology, including two patients with large villous adenomas. Nonpouch ileal mucosa was spared from visually observed adenomas, with only 1 of 48 (2 percent) patients with ileorectal anastomosis adenomas and 0 of 20 (0 percent) younger, precolectomy patients having terminal ileal adenomas. However, microadenomas were present on random biopsy in 4 percent to 5 percent of nonpouch ileum. CONCLUSION: The risk of pouch cancer in familial adenomatous polyposis is unclear, but follow-up periods since surgery remain relatively short. Long-term endoscopic surveillance of familial adenomatous polyposis pouches is thus recommended along with evaluation of potential therapeutic options for pouch adenomas. PMID- 15747077 TI - Eight years experience of high-powered endoscopic diode laser therapy for palliation of colorectal carcinoma. AB - PURPOSE: Endoscopic laser therapy using neodymium: yttrium-aluminum-garnet (Nd:YAG) laser has been shown to be effective in palliating symptoms of obstruction, bleeding, and discharge in patients with colorectal cancer. These patients usually have advanced inoperable disease at presentation or are unfit for surgery. We have used high-powered diode laser to palliate patients with inoperable colorectal cancer since 1994. This study was designed to determine the success rate of high-powered diode laser in palliating inoperable colorectal carcinoma and compare these figures with those published for Nd:YAG laser. METHODS: A retrospective analysis was performed of all patients undergoing high powered diode laser therapy for colorectal carcinoma between June 1994 and October 2002 (inclusive) at St. George's Hospital, London, United Kingdom. Patient's notes and endoscopy records were reviewed to determine the indications for treatment, success of symptom palliation, complications, and survival for each patient. RESULTS: Fifty-seven patients (28 males), with a median age at first treatment of 82 (range, 51-93) years, were identified who had been palliated with high-powered diode laser therapy for colorectal carcinoma. The median number of treatments received by each patient was three (range, 1-16 treatments), with a median interval between treatments of 9.5 (range, 1-25) weeks. Lifelong palliation of symptoms occurred in 51 patients (89 percent). Major complications were two perforations and one hemorrhage, giving an overall complication rate of 5.3 percent. One of the patients who experienced perforation died, giving an overall mortality rate of 1.8 percent for the procedure. The median survival of the 51 patients palliated completely by laser therapy was 8.5 (range, 0.6-52) months, with a probability of survival at 24 months of 15 percent. CONCLUSIONS: High-powered diode laser therapy is an effective method of providing palliation for obstruction, bleeding, and discharge in those patients with inoperable colorectal carcinoma. It produces results comparable to therapy with Nd:YAG laser and the equipment is cheaper, more compact, and portable. PMID- 15747078 TI - Growth in epithelial cell proliferation and apoptosis correlates specifically to the inflammation activity of inflammatory bowel diseases: ulcerative colitis shows specific p53- and EGFR expression alterations. AB - PURPOSE: Epithelial cell turnover related differences between ulcerative colitis, Crohn's colitis, and aspecific colitis are not known yet. METHODS: Totally 345 formalin-fixed, paraffin-embedded biopsy specimens from 33 ulcerative colitis, 26 Crohn's colitis, 30 aspecific colitis, and 10 healthy patients were observed with the TdT-mediated dUTP nick end labeling method and proliferating cell nuclear antigen-, p53-, and epithelial growth factor receptor immunohistochemistry. Because of epithelial growth factor receptor positivity of subepithelial cells epithelial growth factor receptor and CD45, CD68, or CD83 double fluorescence immunohistochemistry were performed on 16 freshly frozen samples from 8 severely active ulcerative colitis and 8 severely active Crohn's colitis patients to describe lamina propria's mononuclear cells, respectively. RESULTS: The epithelial growth factor receptor expression was significantly lower in each inflammatory group compared with normal (P < 0.005) and decreased significantly in mild ulcerative colitis compared with mild Crohn's colitis or aspecific colitis (P < 0.005). Numerous epithelial growth factor receptor and CD45 double positive submucosal mononuclear cells were observed in moderate-severe inflammations. The p53-expression was significantly higher in each inflammatory group compared with normal (P < 0.05). Significant differences were found between mildly, moderately, and severely inflamed samples in ulcerative colitis (P < 0.05) compared with Crohn's colitis or aspecific colitis. Apoptotic/proliferative rates increased significantly in line with the inflammatory process (P < 0.0001/0.05), but the TdT-mediated dUTP nick end labeling and proliferating cell nuclear antigen-labeling characteristics did not show disease type specificity. CONCLUSIONS: Based on our results, the alterations of epithelial growth factor receptor and p53 expression show ulcerative colitis specificity, whereas the rate of epithelial apoptosis and proliferation are determined by the histologic activity of the inflammation. The increased epithelial growth factor receptor expression by the lamina propria's mononuclear cells in inflammation may suggest its role as an autoantigen. PMID- 15747079 TI - Ten-year historic cohort of quality of life and sexuality in patients with rectal cancer. AB - PURPOSE: In various studies, type of surgery, age, and gender had different impact on sexuality and quality of life in patients with rectal cancer. This study was designed to investigate how sexuality and quality of life are affected by age, gender, and type of surgery. METHODS: A total of 516 patients who had undergone surgery for rectal cancer in our department from 1992 to 2002 were included. Within one year after the operation, 117 patients died. Questionnaires were sent to 373 patients 12 to 18 months after surgery. We received quality of life data from 261 patients. Comparisons were made after adjusting age, gender, and type of surgical procedure. RESULTS: For patients receiving abdominoperineal resection sexuality was most impaired. Significant differences were seen in symptom and function scales between males and females. Females reported more distress from the medical treatment insomnia, fatigue, and constipation. Both genders had impaired sexual life; however, males had significantly higher values and felt more distressed by this impairment. Younger females felt more distress through impaired sexuality. In males sexuality was impaired independent of age. Adjuvant therapy had no influence on sexuality but on quality of life one year after surgery. CONCLUSIONS: Assessing quality of life with general and specific instruments is helpful to determine whether patients improved through the treatment. The study showed that gender, age, and type of surgery influence sexuality and that quality of life after surgery for rectal cancer is impacted. Because quality of life is a predictor for complications and survival, availability of such data may help to direct supportive treatment to improve outcome. PMID- 15747080 TI - Association between fecal hydrogen sulfide production and pouchitis. AB - PURPOSE: The beneficial effect of antibiotics in pouchitis suggests that an unidentified fecal bacterial product causes this condition. A candidate compound is hydrogen sulfide, a highly toxic gas produced by certain fecal bacteria, which causes tissue injury in experimental models. We investigated hydrogen sulfide release and sulfate-reducing bacterial counts in pouch contents to determine whether hydrogen sulfide production correlates with pouchitis. METHODS: During incubation at 37 degrees C, the production of hydrogen sulfide, methylmercaptan, carbon dioxide, and hydrogen were studied using fresh fecal specimens obtained from 50 patients with ileoanal pouches constructed after total proctocolectomy for ulcerative colitis (n = 45) or for familial adenomatous polyposis (n = 5). Patients with ulcerative colitis were divided into five groups: a) no history of pouchitis (pouch for at least 2 years; n = 8); b) past episode(s) of pouchitis but no active disease for the previous year (n = 9); c) pouchitis in the past year but presently inactive (n = 9); d) ongoing antibiotic treatment (metronidazole or ciprofloxacin) for pouchitis (n = 11); e) currently suffering from pouchitis (n = 8). RESULTS: Release of hydrogen sulfide when pouchitis was active (6.06 +/- 1.03 micromol g(-1) 4 h(-1)) or had occurred in the past year (4.71 +/- 0.41 pmol g(-1) 4 h(-1)) was significantly higher (P < 0.05) than when pouchitis had never occurred (1.71 +/- 0.43 micromol g(-1) 4 h(-1)) or had been inactive in the past year (2.62 +/- 0.49 micromol g(-1) 4 h(-1)). Antibiotic therapy was associated with very low hydrogen sulfide release (0.68 +/- 0.29 micromol g(-1) 4 h(-1)). Pouch contents from familial adenomatous polyposis patients produced significantly less hydrogen sulfide (0.75 +/- 0.09 micromol g( 1) 4 h(-1)) than did any group of nonantibiotic-treated ulcerative colitis patients. Sulfate-reducing bacterial counts in active pouchitis (9.5 +/- 0.5 log10/g) were significantly higher than in those who never experienced pouchitis (7.38 +/- 0.32 log10/g), and these counts fell dramatically with antibiotic treatment. No statistically significant differences in carbon dioxide and hydrogen were observed among the groups not receiving antibiotics. CONCLUSIONS: Pouch contents of patients with ongoing pouchitis or an episode within the previous year released significantly more hydrogen sulfide than did the contents of patients who never had an attack of pouchitis and those with longstanding inactive disease. The response to therapy with metronidazole or ciprofloxacin was associated with marked reductions in hydrogen sulfide release and sulfate reducing bacteria. These results provide a rationale for additional studies to determine whether the high sulfide production is a cause or effect of pouchitis. The lower hydrogen sulfide production by pouch contents of familial adenomatous polyposis vs. patients with ulcerative colitis suggests a fundamental difference in gut sulfide metabolism that could have implications for the etiology of ulcerative colitis as well as the pouchitis of patients with ulcerative colitis. PMID- 15747081 TI - Volume measurements of the anal sphincter complex in healthy controls and fecal incontinent patients with a three-dimensional reconstruction of endoanal ultrasonography images. AB - OBJECTIVES: The aim of this study was to determine sphincter volume, length, and external anal sphincter thickness in healthy controls and fecal incontinent patients by use of a three-dimensional reconstruction of endoanal ultrasonography images. METHODS: Forty-four controls (15 males, 15 females, and 14 parous females) and 28 incontinent parous females (with and without a sphincter defect) were studied. Internal anal sphincter, external anal sphincter and puborectalis volume, sphincter length, and external anal sphincter thickness were measured. Intraobserver and interobserver variability were assessed. Anal pressure profile was also determined. RESULTS: Internal anal sphincter and external anal sphincter volumes were larger in males than in females (P = 0.001 and P = 0.04), and external anal sphincter volume was smaller in parous females but this was not significant (P = 0.084). Anterior sphincter length was longer in males (P = 0.004) and shorter in parous females (P = 0.06). Males had a larger anterior external anal sphincter thickness (P = 0.018); parity made no difference. Sphincter volumes were not smaller in incontinent females. Incontinent females with a sphincter defect had a shorter anterior sphincter length than that of continent (P = 0.001) and incontinent females without a sphincter defect (P < 0.001). Anterior external anal sphincter thickness was smaller in incontinent females with a sphincter defect (P = 0.006), and posterior and right external anal sphincter thickness was smaller in incontinent females without a sphincter defect (P = 0.02 and P = 0.03). Intraobserver variability was seen for internal anal sphincter volume and sphincter length, but there was no interobserver variability. Correlation between anal pressures and endoanal ultrasonography measurements was poor. CONCLUSIONS: Differences in anal sphincter volumes are seen for gender but not for parity. Fecal incontinence is not associated with loss of sphincter volume. However, anterior sphincter length and external anal sphincter thickness are smaller. PMID- 15747082 TI - Metastatic testicular carcinoma from the colon with clinical, immunophenotypical, and molecular characterization: report of a case. AB - We report a case of testicular metastasis from a colonic adenocarcinoma. The presentation of testicular metastasis, diagnosis, management, and possible modes of spread are reported. In addition to conventional investigations and histopathologic techniques, a molecular study of the testicular metastasis and colon primary, using microsatellite analysis, was performed to confirm the primary origin. Its potential uses are discussed. PMID- 15747083 TI - Outcome of anterior sphincter repair for obstetric injury: comparison of early and late results. AB - INTRODUCTION: Fecal incontinence is commonly caused by structural sphincter damage secondary to obstetric trauma. Anterior sphincter repair achieves reasonable early improvement rates of between 69 and 97 percent. Few series have reported long-term results. This study was designed to evaluate the long-term outcome and examine whether there are any predictive factors that could refine patient selection and predict long-term outcome. METHODS: The case records of all patients who underwent anterior sphincter repair between January 1991 and December 1999 were studied. The patients were sent a questionnaire that asked about preoperative and postoperative and current bowel function, with questions about quality of life and overall satisfaction with the outcome of the procedure. The late outcome after a mean period of 70 months from the operation was compared with the early clinical results. All the preoperative and operative variables were studied to ascertain their significance in predicting success. RESULTS: Ninety-three patients were admitted to the study. Anterior sphincter repair was successful in improving continence in 73 percent of patients. Long-term results were obtained for 62 patients. Seventy percent had objective clinical improvement based on the questionnaire, but only 55 percent considered their bowel control had improved and only 45 percent were satisfied by the operation. Urgency was the most important symptom in determining patient satisfaction; 24 of 26 patients in whom urgency had improved were happy with their outcome. None of the preoperative and operative variables predicted the outcome. CONCLUSIONS: Patients should be warned that complete continence is difficult to achieve and that symptoms tend to deteriorate with time. PMID- 15747084 TI - Evaluation of P-POSSUM in surgery for obstructing colorectal cancer and correlation of the predicted mortality with different surgical options. AB - PURPOSE: This study examined the accuracy of Portsmouth Physiologic and Operative Severity Score for enUmeration of Mortality and Morbidity system (P-POSSUM) in predicting the mortality of patients who underwent operations for obstructing colorectal cancer. It also is attempted to analyze the actual mortality and the predicted P-POSSUM mortality of different surgical options for obstructing left sided cancer. METHODS: Data on patients who underwent surgery for obstructing colorectal cancer during 1998 to 2002 were collected. Mortality predicted by P POSSUM was compared to the actual mortality with the method of linear analysis. The accuracy of using P-POSSUM to predict mortality in this group of patients was assessed by Hosmer and Lemeshow goodness of fit test and Receiver Operator Characteristic curve analysis. The predicted and actual mortality of patients who underwent different surgical options also were analyzed. RESULTS: A total of 160 patients were included in the study and 18 patients died postoperatively. The operative mortality was 11.3 percent. P-POSSUM predicted overall mortality of 15 percent. The observed and predicted mortality was found to have no significant lack of fit (chi-squared = 5.98; degree of freedom = 3; P = 0.11). The area under Receiver Operator Characteristic curve analysis was 0.75. For patients with left sided tumors, P-POSSUM predicted mortality and actual mortality of patients who had resection without anastomosis were both significantly higher than patients with single-stage resection and primary anastomosis (P = 0.044 and 0.011, respectively). CONCLUSIONS: P-POSSUM system is valid for prediction of overall mortality in patients with operations for obstructing colorectal cancer. Estimation of P-POSSUM predicted mortality during operation and its ability to correlate with choice of procedure is an area that is worth further study in emergency colorectal surgery. PMID- 15747087 TI - Degradation of ferric chelate of ethylenediaminetetraacetic acid by bacterium isolated from deep-sea stalked barnacle. AB - Twenty strains of marine bacteria that degrade ferric chelate of ethylenediaminetetraacetic acid (Fe-EDTA) were isolated from among 117 strains collected from a marine environment. Among them strain 02-N-2, which was isolated from stalked barnacle collected from the deep sea in the Indian Ocean, had the highest Fe-EDTA degradation ability and was selected for further study. The strain showed high Fe-EDTA degradation ability at different seawater concentrations. In addition, the intact cells of this strain had the ability to degrade such metal-EDTAs as Ca, Cu, and Mg. The strain was an aerobic, gram variable, rod-shaped organism. The results of various taxonomic studies revealed that the strain had significant similarity to Bacillus jeotgali JCM 10885(T), which was isolated from a Korean traditional fermented seafood, Jeotgal. PMID- 15747088 TI - Genetic analysis of selected strains of eastern oyster (Crassostrea virginica Gmelin) using AFLP and microsatellite markers. AB - Amplified fragment length polymorphisms (AFLPs) and microsatellite markers were used to examine genetic variation and divergence in 4 selected strains (DBH, NEH, FMF, and CTS) and 1 wild population (DBW) of the eastern oyster Crassostrea virginica Gmelin. Eighty-six AFLP markers (from 3 primer pairs) and 5 microsatellite loci were used for the analysis of 30 oysters from each of the 5 populations. Microsatellite loci were considerably more variable than AFLPs. The observed heterozygosity ranged from 0.560 to 0.640 across populations for microsatellites, and from 0.186 to 0.207 for AFLPs. Both Fst and phiPT of microsatellite data and phiPT statistics of AFLP data revealed significant divergence between all pairs of populations. There was no significant reduction in heterozygosity in all 4 selected strains; however, the number of alleles per locus was considerably lower in the selected strains than in the wild population. Two strains subjected to long-term selection for disease resistance shared frequency shifts at a few loci, which deserve further analysis to determine if they are linked to disease-resistance genes. PMID- 15747089 TI - Polymorphism in Nucleotide Sequence of Mitochondrial Intergenic Region in Scleractinian Coral (Galaxea fascicularis). AB - A region of 826 bp that is unlikely to code for a protein, ribosomal RNA, or transfer RNA was identified between the cytochrome b and NADH-ubiquinone oxidoreductase chain 2 loci in the mitochondrial DNA of the scleractinian reef coral Galaxea fascicularis. Nucleotide sequences were determined in a part (625 bp) of this intergenic region in 95 individuals collected at 9 sites in the Ryukyu Archipelago in southwestern Japan. A total of 8 haplotypes were found, and a deletion of 290 bp was found in 3 of them. Significant differences were found in frequencies of the haplotypes at 3 sampling sites. The presence or absence of the deletion was highly correlated with the hard or soft morphotype. The deletion was found in the majority of hard-type colonies, but in a small fraction of soft type individuals. PMID- 15747090 TI - Zona localization of shell matrix proteins in mantle of Haliotis tuberculata (Mollusca, Gastropoda). AB - Organic matrix from molluscan shells has the potential to regulate calcium carbonate deposition and crystallization. Control of crystal growth thus seems to depend on control of matrix protein secretion or activation processes in the mantle cells, about which little is known. Biomineralization is a highly orchestrated biological process. The aim of this work was to provide information about the source of shell matrix macromolecule production, within the external epithelium of the mantle. An in vivo approach was chosen to describe the histologic changes in the outer epithelium and in blood sinus distribution, associated with mantle cells implicated in shell matrix production. Our results characterized a topographic and time-dependent zonation of matrix proteins involved in shell biomineralization in the mantle of Haliotis. PMID- 15747091 TI - Phylogenetic relationships among ten sole species (Soleidae, Pleuronectiformes) from the Gulf of Cadiz (Spain) based on mitochondrial DNA sequences. AB - The entire sequence of the mitochondrial cytochrome b gene and 2 partial sequences of the ribosomal RNA12S and 16S genes have been used to study the molecular phylogeny in 10 species of soles belonging to the genera Solea, Monochirus, Microchirus, Dicologlossa, and Synaptura from the Atlantic waters of the Gulf of Cadiz (Spain). The results obtained by means of different phylogenetic analyses (maximum likelihood, maximum parsimony, and neighbor joining) were quite similar, supporting the monophyly of the Solea species. Nevertheless, they favor the differentiation of Dicologlossa cuneata and Dicologlossa hexophthalma in 2 distinct genera, since the most closely related species to the last one is Microchirus azevia. The fact that M. azevia is also more closely linked to Monochirus hispidus than to its congeneric Microchirus boscanion argues in favor of a taxonomic reorganization of these genera. PMID- 15747092 TI - Molecular cloning of silicatein gene from marine sponge Petrosia ficiformis (Porifera, Demospongiae) and development of primmorphs as a model for biosilicification studies. AB - In some sponges peculiar proteins called silicateins catalyze silica polymerization in ordered structures, and their study is of high interest for possible biotechnological applications in the nanostructure industry. In this work we describe the isolation and the molecular characterization of silicatein from spicules of Petrosia ficiformis, a common Mediterranean sponge, and the development of a cellular model (primmorphs) suitable for in vitro studies of silicatein gene regulation. The spicule of P. ficiformis contains an axial filament composed of 2 insoluble proteins, of 30 and 23 kDa. The 23-kDa protein was characterized, and the full-length cDNA was cloned. The putative amino acid sequence has high homology with previously described silicateins from other sponge species and also is very similar to cathepsins, a cystein protease family. Finally, P. ficiformis primmorphs express the silicatein gene, suggesting that they should be a good model for biosilicification studies. PMID- 15747093 TI - Purification and partial identification of novel antimicrobial protein from marine bacterium Pseudoalteromonas species strain X153. AB - A marine bacterium, X153, was isolated from a pebble collected at St. Anne du Portzic (France). By 16S ribosomal DNA gene sequence analysis, X153 strain was identified as a Pseudoalteromonas sp. close to P. piscicida. The crude culture of X153 was highly active against human pathogenic strains involved in dermatologic diseases, and marine bacteria including various ichthyopathogenic Vibrio strains. The active substance occurred both in bacterial cells and in culture supernatant. An antimicrobial protein was purified to homogeneity by a 4-step procedure using size-exclusion and ion-exchange chromatography. The highly purified P-153 protein is anionic, and sodium dodecylsulfate polyacrylamide gel electrophoresis gives an apparent molecular mass of 87 kDa. The X153 bacterium protected bivalve larvae against mortality, following experimental challenges with ichthyopathogenic Vibrio. Pseudoalteromonas sp. X153 may be useful in aquaculture as a probiotic bacterium. PMID- 15747094 TI - Preparation of single cell detritus from Laminaria saccharina as a hatchery diet for bivalve mollusks. AB - A high-yield technique is described for the elaboration of single cell detritus (SCD) from Laminaria saccharina, based on the sequential action of C1H, enzymes (endoglucanases and cellulases) and 2 bacteria showing a high degree of cellobiotic, proteolytic, and alginolytic activity (CECT 5255 and CECT 5256). Over 85% of dried particles of L. saccharina were transformed into a suspension of free cell and bacterial and detrital particles after 24 hours of bacterial activity with this technique. These particles were less than 20 mum in diameter, constituting a suitable diet for bivalve mollusks. After 72 hours 99% of the total particulate volume consisted of particles less than 20 mum in diameter. Tests of hatchery diets for the seed of clam Ruditapes decussatus revealed increases of 54% and 68% for live weight and length, respectively, when SCD from L. saccharina was used as the sole dietary component compared with a live phytoplankton diet. However, SCD from L. saccharina is not a suitable food for the larvae of R. decussatus. PMID- 15747095 TI - Population structure of tropical abalone (Haliotis asinina) in coastal waters of Thailand determined using microsatellite markers. AB - Three partial genomic libraries were constructed from genomic DNA of the tropical abalone (Haliotis asinina) that was digested with AluI, vortexed/sonicated, and digested with mixed enzyme (AluI, HincII, and RsaI). The libraries yielded 0.02%, 0.42%, and 1.46% positive microsatellite-containing clones, respectively. Eleven clones each of perfect, imperfect, and compound microsatellites were isolated. Ten primer pairs (CUHas1-CUHas10) were analyzed to evaluate their polymorphic level. The numbers of alleles per locus, observed heterozygosity (H0), and expected heterozygosity (He) ranged from 3 to 26 alleles, and varied between 0.27 and 0.85 and between 0.24 and 0.93, respectively. Three microsatellite loci (CUHas2, CUHas3, and CUHas8) were further used for examination of genetic diversity and differentiation of natural H. asinina in coastal waters of Thailand. Genetic variabilities in terms of the effective number of alleles (n(e)), H0, and He were higher in 2 samples from the Gulf of Thailand (n(e)=9.37, 7.66; H0=0.62, 0.78; and He=0.87, 0.86) than those of one sample (n(e)=6.04; H0=0.58; and He=0.62) derived from the Andaman Sea. Assessment of genetic heterogeneity, including allele frequency comparison and pairwise F(ST) analysis, indicated interpopulational differentiation, between natural H. asinina from the Gulf of Thailand and that from the Andaman Sea (P<0.0001). PMID- 15747096 TI - Multislice CT virtual angioscopy of the abdomen. AB - BACKGROUND: Computed tomographic (CT) angiography represents an important clinical tool in the evaluation of vascular disorders. Virtual angioscopy can be reconstructed with volumetric CT data sets. We evaluated the feasibility and clinical value of this application in the assessment of abdominal vessels. METHODS: Data sets of CT angiographic studies obtained with helical (n = 120) and multislice (n = 180) CT scanners were analyzed on a workstation for postprocessing. Vascular evaluation was done on conventional enhanced axial images, three-dimensional reconstructions, and virtual angioscopic images. RESULTS: We made 123 studies in patients without aortic disease. Of the patients evaluated for stent-graft treatment, 63 showed normal patency, seven had partial thrombosis of the stent-graft, five showed total occlusion of the stent-graft, and 10 had leaks. From the 92 remaining CT studies, 63 vascular aneurysms and nine dissections were diagnosed. CONCLUSION: The current technology produces high quality virtual angioscopic images. Although axial and multiplanar views are usually adequate for detecting a vascular disorder, virtual angioscopic views better define anatomic details. PMID- 15747098 TI - Detection of SYT-SSX rearrangements in synovial sarcomas by real-time one-step RT PCR. AB - Synovial sarcomas are aggressive tumors of adolescent and young adults that account for up to 10% of soft tissue sarcomas. Cytogenetically, they are characterized by translocation t(X;18), which is found in more than 95% of tumors. In most cases, it results in fusion of the SYT gene with the SSX1 or SSX2 gene, thus creating SYT-SSX1 or SYT-SSX2 rearrangement. The 2 types of gene fusion have been correlated with histologic variants and prognosis of synovial sarcomas. In this study, we developed a simple and rapid method for the simultaneous detection of SYT-SSX1 and SYT-SSX2 rearrangements by using a LightCycler real-time one-step reverse transcriptase polymerase chain reaction (RT-PCR) technology (Roche). Oligonucleotide probes were designed so that the donor probe would span a fusion point and the acceptor probe would be complementary to the SSX1 sequence but have 2 nucleotide mismatches with SSX2 sequence. Such a design allows simultaneous amplification of 2 types of rearrangement in the same reaction but distinguishes them based on differences in melting temperature detected by melting curve analysis after PCR. With this method, 27 tumors (9 synovial sarcomas and 18 nonsynovial sarcomas) were studied and showed SYT-SSX1 rearrangement in 6 cases and SYT-SSX2 in 3 cases. These results had complete correlation with the finding of conventional RT-PCR and direct sequencing. In conclusion, we have developed a fast, accurate, and simple method for the detection of 2 major types of SYT-SSX rearrangement by using LightCycler RT-PCR and melting curve analysis. PMID- 15747097 TI - Pediatric renal carcinoma associated with Xp11.2 translocations/TFE3 gene fusions and clinicopathologic associations. AB - Renal cell carcinomas (RCCs) are rare in children and studies of their subtypes and clinicopathologic associations are limited to small series. We identified 8 patients with RCC treated at our institution between 1981 and 2003, reviewed their clinicopathologic features, cytogenetics findings, and evaluated the status of TFE3 expression by immunohistochemistry and numerical chromosomal alterations by interphase fluorescent in situ hybridization on paraffin-embedded tissue. These 8 patients (5 female and 3 male) had diploidy, and 5 had morphologic features compatible with the recently described RCC associated with Xp11.2 translocations/TFE3 gene fusions and demonstrated nuclear labeling for TFE3 protein by immunohistochemistry. The translocation was confirmed in 2 of these 5 patients by conventional cytogenetics. One case was a high-grade nonpapillary RCC and the other was compatible with type 2 papillary RCC. Four patients showed at least 1 chromosomal gain including trisomy 7 and/or trisomy 17. None of the tumors from male patients showed evidence of loss of the Y chromosome, but 2 patients showed numerical abnormalities of X chromosome +add(X). Two patients had sickle cell disease, and 1 of these also had stage IV-S neuroblastoma. This study suggests that many cases of RCC in children reported under the terms "papillary" and "clear cell" likely represent Xp11.2 translocation/TFE3 gene fusion associated RCC. It also emphasizes the unusual associations of RCC with neuroblastoma and sickle cell hemoglobinopathy, which need further study. PMID- 15747099 TI - Pediatric inflammatory myofibroblastic tumor with late metastasis to the lung: case report and review of the literature. AB - Inflammatory myofibroblastic tumors (IMTs) are challenging lesions with respect to classification, differential diagnosis, and biologic potential. In children, extrapulmonary IMTs, particularly those from the abdomen or mesentery, are generally aggressive, with frequent local recurrences. There are few documented patients with distant metastasis, and most of these had metastases at presentation or developed metastases within months to a few years. We add to the short list of metastatic extrapulmonary IMTs a pediatric patient in whom the primary lesion was widespread in the abdomen at presentation and metastatic disease to the lung was diagnosed 9 years after the primary resection. We describe the clinical and pathologic features of this patient and review the characteristics of extrapulmonary IMTs with distant metastasis reported in the literature. PMID- 15747100 TI - Autopsy standards of body parameters and fresh organ weights in nonmacerated and macerated human fetuses. AB - Standards for body parameters and organ weights are important tools in fetal and perinatal pathology. Previously there has been only a weak emphasis on the effect of maceration on dimensions and weights. This study provides autopsy standards for body weight, body dimensions, and fresh organ weights for nonmacerated fetuses and for mildly, moderately, and markedly macerated fetuses at 12 to 43 weeks of gestation. Cases were selected from a consecutive series of 1800 fetal and perinatal autopsies. Cases with malformations, hydrops, infection, or chromosomal abnormality, fetuses from multiple births, and infants who lived longer than 24 h were excluded. In each case the maceration was graded and body weight and 4 body dimensions were recorded before organ examination. Organs were weighed immediately and before fixation. Growth curves were fitted according to appropriate mathematical methods and the effects of maceration on each value were tested statistically. We found that weights of the liver, thymus, and spleen markedly decrease with increasing maceration. The weights of the lungs, kidneys, and adrenals decreased modestly, whereas weights of the heart and brain changed only slightly. Body length increased slightly with maceration, whereas body weight and head circumference were unaffected. User-friendly charts and tables of mean values and standard deviations for nonmacerated and macerated fetuses are provided. PMID- 15747101 TI - Ovarian mature cystic teratoma exhibiting well-differentiated cerebellum in a 14 year-old female. PMID- 15747102 TI - Renal metanephric adenoma with previously unreported cytogenetic abnormalities: case report and review of the literature. AB - We report a case of a renal metanephric adenoma in a 10-year-old boy, in which cytogenetic analysis showed a balanced translocation, t(9;15)(p24;q24) and a balanced paracentric inversion of chromosome 12, inv(12)(q13q15). Immunohistochemically, the tumor showed diffuse reactivity for cytokeratin AE1/AE3, CAM5.2, CD57, and WT1; patchy reactivity for CD56; and focal reactivity for cytokeratin 7, epithelial membrane antigen, and CD10. Tumor cells were entirely nonreactive for alpha-methyl acyl coenzyme A racemase. Published cytogenetic data for metanephric adenomas are limited, and this is the first report of these cytogenetic abnormalities. The involvement of the chromosome region 9p24 is particularly interesting because of the recent identification of a tumor suppressor gene, KANK (kidney ankyrin repeat-containing protein), at this locus. PMID- 15747104 TI - Mucocutaneous squamous papilloma with reactive lymphoid hyperplasia in two patients with focal dermal hypoplasia. PMID- 15747103 TI - Histologic analysis of gonadal tissue in patients with Ullrich-Turner syndrome and derivative Y chromosomes. AB - To identify patients who had Ullrich-Turner syndrome (UTS) and were at risk for gonadoblastoma or associated germ cell tumors, molecular genetic analysis was carried out to detect Y chromosomal sequences. From peripheral blood samples of 5 patients who had cytogenetically confirmed UTS, genomic DNA was extracted and screened for Y chromosomal sequences by polymerase chain reaction. The morphology of the gonadal tissues was compared with results from polymerase chain reaction. Three phenotypic females showed UTS mosaicism with normal X chromosome accompanied by Y chromosomal material, and 2 patients showed marker chromosomes. Molecular analysis represented loci PABY, SRY, ZFY, TSPY, DYZ3, DYZ1 DXYS, 19Y, DYS-273, DYS-148, DYS218, DYS224, and DYZ1. Three patients showed gonadal tumors (1 with unilateral gonadoblastoma, 1 with unilateral dysgerminoma, and 1 patient had both tumors in 1 gonad). Molecular genetic screening for Y chromosomal sequences may be useful as an additional tool for the identification of patients at risk for a gonadal tumor. Careful, complete processing, including step sectioning, of the gonadectomy specimens to detect small lesions is recommended. PMID- 15747105 TI - Management of acoustic neuroma in the only hearing ear. AB - Patients with acoustic neuroma in their only hearing ear are not frequently seen in clinical practice. Managing this group of patients is a challenge to both the patient and surgeon. In this study we report on five cases of acoustic neuroma in an only hearing ear. Our decision for conservative management of those patients with regular follow-up using auditory brain stem response and magnetic resonance imaging is discussed. Other management options currently available are reviewed as well. PMID- 15747106 TI - Intratympanic dexamethasone with hyaluronic acid in the treatment of idiopathic sudden sensorineural hearing loss after failure of intravenous steroid and vasoactive therapy. AB - The purpose of this prospective study was to test whether intratympanic application of dexamethasone/hyaluronic acid improves hearing outcome in patients with pantonal idiopathic sudden sensorineural hearing loss (ISSHL), in patients with sudden deafness or sudden profound SHL and in patients with predominant high frequency ISSHL who are refractory to intravenous steroid and vasoactive therapy. The study took place in an academic tertiary referral hospital involving 21 patients with pantonal ISSHL, 10 patients with sudden deafness or sudden profound SHL and 9 patients with a high-frequency ISSHL. Intratympanic dexamethasone/hyaluronic acid was administered in the affected ear. Hearing was evaluated by means of standard pure-tone audiometry. The differences between pure tone hearing thresholds by air conduction before intravenous therapy and before the beginning of the intratympanic therapy, as well as before and after intratympanic therapy, were calculated. Statistical analysis was performed by means of the Wilcoxon's test for paired samples. Intratympanic injection of dexamethasone/hyaluronic acid results in a significant global (pantonal) improvement in hearing in patients with pantonal ISSHL. It also effects improvement in hearing at selected frequencies (namely at 1.5 and 3 kHz) in patients with a predominant high-frequency ISSHL and at selected frequencies (namely at 0.5, 0.75 and 1 kHz) in patients with sudden deafness or sudden profound SHL. Neither systemic nor local side effects were observed. Intratympanic administration of dexamethasone/hyaluronic acid provides a safe and efficacious therapeutic option for the treatment of patients with pantonal and high-frequency ISSHL who don't respond to intravenous steroid and vasoactive therapy. PMID- 15747107 TI - Comment to: Borch-Johnsen K, Colagiuri S, Balkau B et al. (2004) Creating a pandemic of prediabetes: the proposed new diagnostic criteria for impaired fasting glycaemia. Diabetologia 47:1396-1402. PMID- 15747108 TI - Increased aortic stiffness is persistent in type 1 diabetic women: a follow-up study. AB - AIMS/HYPOTHESIS: We have previously reported that women, not men, with type 1 diabetes have increased aortic stiffness. Increased arterial stiffness may explain why diabetic women have a particularly high risk of developing cardiovascular complications. We have now followed up our previously investigated patients after 7 years, with a view to evaluating whether the sex difference was persistent, and also evaluating the degree of progression with time and the relationship between stiffness versus intima media thickness of the aorta. METHODS: Stiffness (beta) of the abdominal aorta (echo-tracking sonography) and intima media thickness (B-mode ultrasound) were assessed in 23 women and 19 men with type 1 diabetes and compared with matched healthy individuals. RESULTS: At follow-up, aortic stiffness was still higher (60%) (p=0.0016) in diabetic than in control women, whereas there was no similar difference (p=0.4) between diabetic and control men. No progression of stiffness had occurred over the 7 years. At follow-up, the intima media thickness was increased and the internal diameter of the aorta was decreased in diabetic men and women without any sex-related difference. CONCLUSIONS/INTERPRETATION: The increased aortic stiffness that affects type 1 diabetic patients seems to be an early event that soon reaches a plateau without any further increase. Increased aortic stiffness in type 1 diabetic women seems to be a sex-specific functional disorder unrelated to the degree of underlying atherosclerosis. PMID- 15747109 TI - Human triglyceride-rich lipoproteins impair glucose metabolism and insulin signalling in L6 skeletal muscle cells independently of non-esterified fatty acid levels. AB - AIMS/HYPOTHESIS: Elevated fasting and postprandial plasma levels of triglyceride rich lipoproteins (TGRLs), i.e. VLDL/remnants and chylomicrons/remnants, are a characteristic feature of insulin resistance and are considered a consequence of this state. The aim of this study was to investigate whether intact TGRL particles are capable of inducing insulin resistance. METHODS: We studied the effect of highly purified TGRLs on glycogen synthesis, glycogen synthase activity, glucose uptake, insulin signalling and intramyocellular lipid (IMCL) content using fully differentiated L6 skeletal muscle cells. RESULTS: Incubation with TGRLs diminished insulin-stimulated glycogen synthesis, glycogen synthase activity, glucose uptake and insulin-stimulated phosphorylation of Akt and glycogen synthase kinase 3. Insulin-stimulated tyrosine phosphorylation of IRS-1, and IRS-1- and IRS-2-associated phosphatidylinositol 3-kinase (PI3K) activity were not impaired by TGRLs, suggesting that these steps were not involved in the lipoprotein-induced effects on glucose metabolism. The overall observed effects were time- and dose-dependent and paralleled IMCL accumulation. NEFA concentration in the incubation media did not increase in the presence of TGRLs indicating that the effects observed were solely due to intact lipoprotein particles. Moreover, co-incubation of TGRLs with orlistat, a potent active-site inhibitor of various lipases, did not alter TGRL-induced effects, whereas co incubation with receptor-associated protein (RAP), which inhibits interaction of TGRL particles with members of the LDL receptor family, reversed the TGRL-induced effects on glycogen synthesis and insulin signalling. CONCLUSIONS/INTERPRETATION: Our data suggest that the accumulation of TGRLs in the blood stream of insulin resistant patients may not only be a consequence of insulin resistance but could also be a cause for it. PMID- 15747110 TI - Insulin resistance in non-diabetic patients with non-alcoholic fatty liver disease: sites and mechanisms. AB - AIMS/HYPOTHESIS: Non-alcoholic fatty liver disease (NAFLD) has been associated with the metabolic syndrome. However, it is not clear whether insulin resistance is an independent feature of NAFLD, and it remains to be determined which of the in vivo actions of insulin are impaired in this condition. METHODS: We performed a two-step (1.5 and 6 pmol min(-1) kg(-1)) euglycaemic insulin clamp coupled with tracer infusion ([6,6-2H2]glucose and [2H5]glycerol) and indirect calorimetry in 12 non-obese, normolipidaemic, normotensive, non-diabetic patients with biopsy proven NAFLD and six control subjects. RESULTS: In NAFLD patients, endogenous glucose production (basal and during the clamp) was normal; however, peripheral glucose disposal was markedly decreased (by 30% and 45% at the low and high insulin doses, respectively, p<0.0001) at higher plasma insulin levels (p=0.05), due to impaired glucose oxidation (p=0.003) and glycogen synthesis (p<0.001). Compared with control subjects, glycerol appearance and lipid oxidation were significantly increased in NAFLD patients in the basal state, and were suppressed by insulin to a lesser extent (p<0.05-0.001). The lag phase of the in vitro copper-catalysed peroxidation of LDL particles was significantly shorter in the patients than in the control subjects (48+/-12 vs 63+/-13 min, p<0.04). Lipid oxidation was significantly related to endogenous glucose production, glucose disposal, the degree of hepatic steatosis, and LDL oxidisability. CONCLUSIONS/INTERPRETATION: Insulin resistance appears to be an intrinsic defect in NAFLD, with the metabolic pattern observed indicating that adipose tissue is an important site. PMID- 15747111 TI - Genome-wide scans for heritability of fasting serum insulin and glucose concentrations in hypertensive families. AB - AIMS/HYPOTHESIS: The heritability of fasting serum insulin and glucose concentrations in non-diabetic members of multiplex hypertensive families is unknown. METHODS: We calculated the familial aggregation of fasting serum glucose and insulin concentrations and performed a genome-wide scan to assess whether quantitative trait loci contribute to these phenotypes in 2,412 non-diabetic individuals from 1,030 families enrolled in the Hypertension Genetic Epidemiology Network (HyperGEN) in the Family Blood Pressure Program. RESULTS: The heritability (+/-SE) of fasting serum insulin was 0.47+/-0.085 in European Americans and 0.28+/-0.08 in African Americans (p<0.0001 for both), after adjusting for age, sex, and BMI. A genome-wide scan for fasting serum insulin yielded a maximum log of the odds (LOD) score of 2.36 on chromosome 5 at 20 cM (p=0.0004) in European Americans, and an LOD score of 2.28 on chromosome 19 at 11 cM (p=0.0004) in African Americans. The heritability of fasting serum glucose was 0.5109+/-0.08 in the former and 0.29+/-0.09 in the latter (p<0.0003 for both) after adjusting for age, sex and BMI. A genome-wide scan for fasting serum glucose revealed a maximum LOD score of 2.07 on chromosome 5 at 26 cM (p=0.0009) in European Americans. CONCLUSIONS/INTERPRETATION: These analyses demonstrate the marked heritability of fasting serum insulin and glucose concentrations in families enriched for the presence of members with hypertension. They suggest that genes associated with fasting serum insulin concentration are present on chromosomes 19 and 5, and that genes associated with fasting serum glucose concentration are on chromosome 5, in families enriched for hypertension. PMID- 15747112 TI - Polycystin-2--an intracellular or plasma membrane channel? AB - The intracellular location of polycystin-2 is a hotly debated topic in the field of polycystic kidney disease. Two not necessarily mutually exclusive hypotheses state that polycystin-2 is located in the endoplasmic reticulum or in the plasma membrane, respectively. Although a variety of techniques have been employed to prove one or the other location, no definite consensus has been reached yet. It is generally acknowledged, however, that the COOH-terminus of polycystin-2 contains a retention signal for the endoplasmic reticulum. Another facet has been added to the discussion due to the fact that many genes mutated in patients with cystic kidney diseases, among them PKD2, encode proteins which have been detected in primary cilia. Since there is no evidence that the endoplasmic reticulum extends into the primary cilium, polycystin-2 has to reach the plasma membrane at least in this case. An unbiased approach towards elucidating the physiological location of polycystin-2 would involve the characterization of its intracellular trafficking. Using the COOH-terminus of polycystin-2 in a two-hybrid screen, my group has identified a novel coiled-coil protein which we call PIGEA-14 (polycystin-2 interactor, Golgi- and endoplasmic reticulum-associated protein with a molecular weight of 14 kDa). PIGEA-14 also interacts with GM130, a protein associated with the Golgi matrix, and may therefore represent one important component of the trafficking machinery for polycystin-2. PMID- 15747114 TI - An analysis of avidin, biotin and their interaction at attomole levels by voltammetric and chromatographic techniques. AB - The electroanalytical determination of avidin in solution, in a carbon paste, and in a transgenic maize extract was performed in acidic medium at a carbon paste electrode (CPE). The oxidative voltammetric signal resulting from the presence of tyrosine and tryptophan in avidin was observed using square-wave voltammetry. The process could be used to determine avidin concentrations up to 3 fM (100 amol in 3 microl drop) in solution, 700 fM (174 fmol in 250 microl solution) in an avidin modified electrode, and 174 nM in a maize seed extract. In the case of the avidin modified CPE, several parameters were studied in order to optimize the measurements, such as electrode accumulation time, composition of the avidin modified CPE, and the elution time of avidin. In addition, the avidin-modified electrode was used to detect biotin in solution (the detection limit was 7.6 pmol in a 6 mul drop) and to detect biotin in a pharmaceutical drug after various solvent extraction procedures. Comparable studies for the detection of biotin were developed using HPLC with diode array detection (HPLC-DAD) and flow injection analysis with electrochemical detection, which allowed biotin to be detected at levels as low as 614 pM and 6.6 nM, respectively. The effects of applied potential, acetonitrile content, and flow rate of the mobile phase on the FIA-ED signal were also studied. PMID- 15747115 TI - Lichen-based biosensor for the determination of benzene and 2-chlorophenol: microcalorimetric and amperometric investigations. AB - Preliminary microcalorimetric studies have been performed to analyse the response of a whole epiphytic lichen tissue (Evernia prunastri) to 2-chlorophenol (2Cl phi), a pollutant of oil mill waste-water, in order to evaluate whether the tissue might be used to assess the toxic characteristics of polluted waters. The obtained results (lichen viability expressed in hours, enthalpy variations for the 2Cl-phi/lichen interactions) were used to create a lichen-based biosensor that uses an amperometric oxygen electrode (a Clark electrode) as a transducer. The lichen catalyses aromatic ring cleavage (via pyrocatechase enzymes present in the lichen), and transforms aromatic substances like 2Cl-phi into muconic acid (C6H6O4). Following a full electroanalytical characterisation, the performance of the proposed lichen biosensor was compared to that of a biosensor based on Pseudomonas putida cells, which was originally constructed to monitor benzene in different matrices (water, air, petrol and oil) and was tested in our laboratory previously. PMID- 15747113 TI - The epithelial calcium channels TRPV5 and TRPV6: regulation and implications for disease. AB - The epithelial Ca(2+) channels TRPV5 and TRPV6 represent a new family of Ca(2+) channels that belongs to the superfamily of transient receptor potential channels. TRPV5 and TRPV6 constitute the apical Ca(2+) entry mechanism in active Ca(2+) transport in kidney and intestine. The central role of TRPV5 and TRPV6 in active Ca(2+) (re)absorption makes it a prime target for regulation to maintain Ca(2+) balance. This review covers the hormonal regulation, interaction with accessory proteins and (patho)physiological implications of these epithelial Ca(2+) channels. PMID- 15747116 TI - Determining pirimiphos-methyl in durum wheat samples using an acetylcholinesterase inhibition assay. AB - An electrochemical assay used for detecting acetylcholinesterase (AChE) inhibitors has been optimised to detect pirimiphos-methyl in durum wheat. Pirimiphos-methyl is a phosphothionate insecticide and so it needs to be transformed into the corresponding oxo form to act as an effective AChE inhibitor. The inhibition assay was based on the electrochemical detection of the product of AChE, choline, via choline oxidase biosensors obtained with Prussian Blue modified screen printed electrodes. The procedure for the oxidation of pirimiphos methyl via N-bromosuccinimide (NBS) and AChE inhibition was optimised for reagent concentrations and inhibition time in a buffer solution. A calibration of the pirimiphos-methyl (25-1,000 ng/ml) was obtained in the buffer. The intra-electrode CV ranged between 1.6 and 15.0, whereas the inter-electrode CV ranged between 4.6 and 16.0. The detection limit (LOD) was 38 ng/ml, and the I(50%) was 360 ng/ml. The assay conditions were then re-optimised to work with durum wheat extracts, and the calibrations were obtained under different experimental conditions, such as sample pretreatment (milled or whole grains) and extract concentration. The calibrations were slightly affected by the sample matrix, resulting in an increased LOD (65-133 ng/ml) and I(50%) (640-1,650 ng/ml). The LOD found for the sample, determined under optimal conditions, was 3 mg/kg. Spiked samples were prepared at the EU regulated level (5 mg/kg) and analysed with the optimised protocol, resulting in an average recovery of 70.3%. PMID- 15747118 TI - Kinetics of Bifidobacterium longum ATCC 15707 fermentations: effect of the dilution rate and carbon source. AB - The effect of the dilution rate on biomass and product synthesis in fermentations of glucose, fructose and a commercial mixture of fructooligosaccharides (FOS) by Bifidobacterium longum ATCC 15707 was studied. Kinetic parameters (maximum specific growth rate, Monod constant, maintenance, and yield coefficients) in the mathematical model of the fermentation were estimated from experimental data. In the FOS mixture fermentations, approximately 12% of the total reducing sugars (mainly fructose) in the feed were not metabolized by the bacterium. In fermentations of fructose and the FOS mixture, biomass concentration increased as the dilution rate increased and, once maximum values were reached [3.90 (D=0.20 h(-1)) and 2.54 g l(-1) (D=0.15 h(-1)), respectively], decreased rapidly as the culture was washed out. Formic acid was detected at low dilution rates in glucose and fructose fermentations. The main products in fermentations of the three carbon sources were lactic and acetic acids. Average values of the molar ratio between acetic and lactic acids of 1.18, 1.21 and 0.83 mol mol(-1) were obtained in glucose, fructose and FOS mixture fermentations, respectively. In batch fermentations carried out without pH control this molar ratio was lower than 1.5 only when fructose was used as the carbon source. PMID- 15747117 TI - Characterization of the peptide-binding specificity of Mamu-A*11 results in the identification of SIV-derived epitopes and interspecies cross-reactivity. AB - The SIV-infected Indian rhesus macaque is the most established model of HIV infection, providing insight into pathogenesis and a system for testing novel vaccines. However, only a limited amount of information is available regarding the peptide-binding motifs and epitopes bound by their class I and class II MHC molecules. In this study, we utilized a library of over 1,000 different peptides and a high throughput MHC-peptide binding assay to detail the binding specificity of the rhesus macaque class I molecule Mamu-A*11. These studies defined the fine specificity of primary anchor positions, and dissected the role of secondary anchors, for peptides of 8-11 residues in length. This detailed information was utilized to develop size-specific polynomial algorithms to predict Mamu-A*11 binding capacity. Testing SIVmac239-derived Mamu-A*11 binding peptides for recognition by peripheral blood mononuclear cells (PBMC) from Mamu-A*11-positive, SIV-infected macaques, identified five novel SIV-derived Mamu-A*11 epitopes. Finally, we detected extensive cross-reactivity at the binding level between Mamu A*11 and the mouse H-2 class I molecule Kk. Further experiments revealed that three out of four Mamu-A*11 binding peptides which bound Kk and were immunogenic in Kk mice were also recognized in Mamu-A*11-infected macaques. This is the first detailed description of mouse-macaque interspecies cross-reactivity, potentially useful in testing novel vaccines in mice and macaques. PMID- 15747119 TI - Recent advances in hereditary hemochromatosis. AB - Hereditary hemochromatosis, a very common genetic defect in the Caucasian population, is characterized by progressive tissue iron overload which leads to irreversible organ damage if it is not treated in a timely manner. Recent developments in the field of molecular medicine have radically improved the understanding of the physiopathology and diagnosis of this disease. However, transferrin saturation and serum ferritin are still the most reliable tests for identifying subjects with hereditary hemochromatosis. Therapeutic phlebotomy is the mainstay of the treatment of this disease and the life expectancy of these patients is similar to that of the normal population if phlebotomy is started before the onset of irreversible organ damage. In this review we discuss the genetics, pathophysiology, diagnosis, clinical features, and management of hereditary hemochromatosis. PMID- 15747120 TI - Modified ProMACE-MOPP hybrid regimen with moderate-dose methotrexate for patients with primary CNS lymphoma. AB - The object of this study was to assess the estimation of 2- and 5-year overall survival and tumor response and the frequency and severity of treatment morbidity with a modified ProMACE-MOPP hybrid protocol in patients with primary CNS lymphoma (PCNSL). Thirty-two immunocompetent patients were treated with a regimen of pirarubicin, cyclophosphamide, etoposide, vincristine, and methotrexate (500 mg/m(2)) administered in 21-day cycles. Intraventricular 10 mg of methotrexate was given for eight cycles once a week. Patients received 20 Gy of whole brain radiotherapy after three cycles of chemotherapy. A single cycle of chemotherapy was repeated every 4 months for 2 years. Older patients (aged >60) received a reduced dose of chemotherapeutic agents. Eighteen patients were followed up with neuroimaging and neuropsychological assessments for evidence of CNS toxicity. Sixteen patients completed the regimen as planned. The response rate was 87.5% after the initial chemoradiotherapy. The cumulative survival and progression-free survival rates at 5 years were 56 and 31%, respectively. The median survival time was 68 months. The median progression-free survival time was 39 months. Toxicity included grade 3 or 4 leukopenia in 33% of the cycles administered. There were eight grade 3 or 4 pulmonary toxicities. There were three deaths during chemotherapy: one as a result of sepsis and two of pneumonitis. Three patients (25%) experienced delayed neurologic toxicity while on the complete regimen. Maintaining the dose of methotrexate while adding chemotherapeutic agents improved disease control and overall survival in patients with PCNSL, but early toxicity and delayed neurotoxicity are still a risk of this approach. PMID- 15747121 TI - [Cystoid macular edema in uveitis]. AB - Cystoid macular edema (CME) is the most frequent cause of visual deterioration in uveitis patients. Intraocular inflammation disturbs the blood-retina barrier and leads to retinal edema. The basis of successful treatment is the anti inflammatory and immunosuppressive therapy of uveitis. Restoration of the blood retina barrier is mediated by corticosteroids and nonsteroidal anti-inflammatory agents. Resorption of extracellular fluid is improved by systemic carboanhydrase inhibitors. Despite aggressive therapy loss of visual acuity is frequent. Therefore, early diagnosis of CME and initiation of treatment, even if visual acuity is not yet impeded, is mandatory. PMID- 15747123 TI - [Evaluation of focal arteriolar narrowing of retinal arterioles in glaucoma]. AB - PURPOSE: Vascular risk factors seem to play a role in the pathogenesis of glaucoma. This study was performed to compare the prevalence of focal arteriolar narrowing in glaucoma patients and normals. MATERIAL AND METHODS: Stereoscopic optic disc photographs of 40 normal subjects and 14 primary open-angle glaucoma (POAG) patients were reviewed independently in a masked fashion by two graders. Focal arteriolar narrowing within one disc diameter from the rim edge was evaluated based on two different methods: narrowing present if (1) the arteriole was wider distal to the narrowing and (2) if the arteriole was wider both distal and proximal to the narrowing. RESULTS: With both methods, focal arteriolar narrowing was significantly higher in glaucoma versus normal eyes. Focal arteriolar narrowing was observed with definition 1 in 35.0% of normals and 71.4% of POAG patients and with definition 2 in 12.5% of normals and 42.9% of POAG patients. CONCLUSIONS: Focal arteriolar narrowing is more frequent in glaucoma patients than in normals. Independent of the method for assessing arteriolar narrowing, however, the diagnostic value of arteriolar narrowing seems limited due to the high incidence in normals. PMID- 15747124 TI - Emergent embolisation to control severe haematuria in Wilms' tumour. AB - Wilms' tumour commonly presents with an abdominal mass and gross haematuria. Here, we present the novel application of paediatric renal arterial embolisation to control life-threatening haematuria in Wilms' tumour. PMID- 15747122 TI - [Uveitis in conjunction with rheumatological diseases in childhood]. AB - Children with juvenile chronic arthritis are at risk to develop intraocular inflammation depending on the type of arthritis. The pathogenic mechanisms are unclear; however, an association with antinuclear antibodies is well known. In particular young girls with oligoarticular onset of arthritis are affected most often. Regular ophthalmologic examinations should allow early diagnosis and effective therapy. Complications such as synechiae, cataract, or macula edema are seen especially in uveitis patients with late diagnosis and insufficient anti inflammatory therapy. Better therapeutic regimens have led to a better overall prognosis of intraocular inflammation in recent years. PMID- 15747125 TI - Fibromyxoid sarcoma in a 4-year-old boy: case report and review of the literature. AB - The authors report on the case of a low-grade fibromyxoid sarcoma (LGFMS) with giant rosettes in a 4-year-old boy. The tumor arose in the paravertebral region and had infiltrated the vertebral canal between L2 and S1. A review of the literature indicates that this is one of the youngest patients diagnosed with a tumor of this type, and spinal involvement had never been reported with LGFMS. PMID- 15747126 TI - A steroid-refractory ulcerative colitis revealing Epstein-Barr virus/cytomegalovirus-positive colonic lymphoma. PMID- 15747127 TI - V-Y advancement flap closures for complicated pilonidal sinus disease. AB - BACKGROUND AND AIMS: Hair entry into the midline is the generally accepted theory in pilonidal sinus formation. This theory is also involved in the pathogenesis of the recurrence after different types of operations for pilonidal sinus disease. The relationship of the suture lines and the midline was evaluated in this study. PATIENT AND METHODS: Thirty-four patients with stage 4 or recurrent pilonidal sinus disease who underwent V--Y advancement flap closure were randomized into two groups: vertical suture line unrelated to midline (VLUM, n=18) and vertical suture line related to midline (VLRM, n=16). RESULTS: None of the patients with or without a suction drain developed flap necrosis, local haematomas or seromas. The median length of the postoperative hospital stay was 3 days for theVLRM group and the VLUM group (p>0.05). The median follow-up period was 32 months. During the follow-up period, two recurrences were determined 22 and 15 months after operation in the VLRM group. Recurrences were situated in the scar tissue over the midline. INTERPRETATION AND CONCLUSION: Routine application of suction drains is not recommended after V--Y advancement flap closure. Avoidance of the relationship of the postoperative suture line and the midline, if possible, offers a safe method of definitive reconstruction. PMID- 15747129 TI - Cracks in the shell--zooming in on eggshell formation in the human parasite Schistosoma mansoni. AB - Schistosomiasis, currently the second most common parasitic disease of humans in tropical regions is caused by the eggs of trematode worms of the genus Schistosoma. Understanding egg formation and specifically the synthesis of the eggshell comprises, consequently, a promising starting point to cure and prevent the disease. To shed light on the genetics of the latter process, we analysed the three known S. mansoni eggshell proteins P14, P19 and P48 against the background of the species' inferred proteome and of eggshell proteins identified in other trematode species. Our results suggest that eggshell formation in Schistosoma involves a multitude of different proteins organised in currently three distinct protein families (P14, P48 and P34 eggshell protein family). The first two families are of simple structure. Their respective members share a substantial degree of sequence similarity and are, to date, observed only in the genus Schistosoma. In contrast, the P34 family of eggshell proteins is complex. Its in part highly diverged members share only a conserved motif of 67-aa length on average and are detected in various trematode species. The resulting widespread occurrence of this protein motif suggests an important role during eggshell formation in trematodes. Screening more than 7,000 putative proteins of S. mansoni, we could identify six new members of the P34 protein family that are likely to be involved in eggshell formation in this species. PMID- 15747128 TI - Choice of either beta-catenin or Groucho/TLE as a co-factor for Xtcf-3 determines dorsal-ventral cell fate of diencephalon during Xenopus development. AB - Co-repressor Groucho/Transducin-Like Enhancer of split (TLE) interacts with transcription factors that are expressed in the central nervous system (CNS), and regulates transcriptional activities. In this study, we examined the contribution of Groucho/TLE to CNS development in Xenopus. The functional inhibition of Groucho/TLE using the WRPW motif as a competitor resulted in the conversion of the ventral cell into the dorsal fate in the prospective diencephalon. We also found that the neural plate was expanded laterally without inhibiting neural crest development. In tailbud, the disturbance of trigeminal ganglion development was observed. These observations allow us to conclude that Groucho/TLE plays important roles in the induction and patterning of distinct CNS territories. We found that Xtcf-3 is involved in some of the patterning in these territories. We generated the variant of Xtcf-3, Xtcf-3BDN-, which is suspected to interfere with the interaction between endogenous Groucho/TLE and Xtcf-3. The transcriptional activation of the Xtcf-3-target genes in response to endogenous Wnt/beta-catenin signaling by the overexpression of Xtcf-3BDN- led to a reduction of the ventral diencephalon. This result indicates that transcriptional repression by the Groucho/TLE-Xtcf-3 complex is important for ventral diencephalon patterning. This idea is supported by the finding that the overexpression of the dominant-negative form of Xtcf-3 or axil causes the expansion of the ventral diencephalon. Based on these data, we propose that the localized activation of Wnt/beta-catenin signaling, which converts Tcf from a repressor to an activator, is required for the establishment of dorsal-ventral patterning in the prospective diencephalon. PMID- 15747131 TI - Malignant ectomesenchymoma of the cerebrum. Case report and discussion of therapeutic options. AB - Malignant ectomesenchymoma is a rare tumour that contains both ectodermal and mesenchymal elements. Only three patients with a manifestation in the cerebrum and clinicopathological data have been reported until now. We present a patient with an intracerebral ectomesenchymoma, review the literature and discuss currently available therapeutic options. In a 10-year-old girl, a left suprasellar temporo-parieto-occipitally localised tumour was diagnosed. The tumour was completely excised macroscopically in two surgical sessions. For the mesenchymal part of the tumour she subsequently underwent multidrug chemotherapy followed by radiation therapy. Considering the neuroectodermal element of the tumour, radiotherapy was applied to the craniospinal axis with a local boost. Therapy was tolerated well without any severe side effects. Six years from diagnosis, the patient is alive without a tumour relapse. CONCLUSION: Due to the sparcity of reported cases with malignant ectomesenchymoma, the role of adjuvant therapy is unclear. Multimodal therapy may be able to improve outcome. PMID- 15747132 TI - Prospective surveillance of hospitalisations associated with varicella-zoster virus infections in children and adolescents. AB - Our goal was to determine the epidemiology of severe varicella-zoster virus (VZV) infections in hospitalised paediatric patients. Admissions associated with VZV infection of patients aged 0-16 years were reported by all 38 paediatric units in Switzerland to the Swiss Paediatric Surveillance Unit (SPSU) during 3 consecutive years (4/2000-3/2003). We verified completeness of reporting by capture-recapture analysis with patient records identified by ICD-10 codes. Outcome of illness was assessed 6 months after hospitalisation. A total of 335 cases (235 identified by SPSU reports, 100 by ICD-10 code) were included in this study. Mean age of patients was 4.1 years (median 3.5 years, range 0-16 years); 54% were male. Some 293 (87%) patients presented with chickenpox, 42 (13%) with herpes zoster and 291 (87%) patients were not immunocompromised. A total of 319 complications occurred in 237 (71%) patients: secondary bacterial infections (n =109); central nervous system involvement (n =76); VZV pneumonitis (n =7); others (n =127). Eleven (3%) patients required intensive care and three died. On follow-up, 303 (96%) of 315 patients had completely recovered; sequelae were present in 12 (4%) patients. The calculated hospitalisation rate was 13 per 10(4) cases. CONCLUSION: This study describes a sizeable hospitalisation and complication rate of varicella-zoster virus infections and provides a solid basis for future immunisation recommendations in Switzerland. PMID- 15747130 TI - Comparative analysis of Wingless patterning in the embryonic grasshopper eye. AB - The signaling factor Wingless regulates multiple steps during the postembryonic development of the Drosophila eye. To obtain insight into the molecular regulation of embryonic eye development in primitive insects, we studied the expression of wg and genes projected to interact with wg in the grasshopper Schistocerca americana. We find that the dynamic and complex expression of wg in the early grasshopper procephalon results in three paired expression domains with relevance to eye primordium development. By comparison with Drosophila, these domains are compatible with a conserved function of wg during anteroposterior and dorsoventral axis formation by repression of retinal differentiation and stimulation of tissue proliferation. This is further supported by the expression of grasshopper orthologs of the retina determination genes sine oculis and eyes absent, and by inhibition of retina differentiation in grasshopper eye primordia cultured with LiCl. Surprisingly, the expression of wg and the grasshopper orthologs of pannier, fringe, Delta, and Iroquois complex is inconsistent with induction of midline centered Notch signaling activity, which is essential for Drosophila retina development. Similarly substantial evolutionary divergence is found concerning the control of retina versus head epidermis specification. The transcription factor Extradenticle (Exd), which cooperates with wg in specifying the Drosophila head epidermis, is not detected outside the labral and antennal primordia in the embryonic grasshopper head. Our results, which provide the first insight into the molecular control of eye primordium formation in primitive insects, suggest substantial modification of this process during the evolution of the Drosophila mode of postembryonic eye development. PMID- 15747133 TI - The activation mechanism of human porphobilinogen synthase by 2-mercaptoethanol: intrasubunit transfer of a reserve zinc ion and coordination with three cysteines in the active center. AB - Human porphobilinogen synthase [EC.4.2.1.24] is a homo-octamer enzyme. In the active center of each subunit, four cysteines are titrated with 5,5'-dithiobis(2 nitrobenzoic acid). Cys(122), Cys(124) and Cys(132) are placed near two catalytic sites, Lys(199) and Lys(252), and coordinate a zinc ion, referred to as "a proximal zinc ion", and Cys(223) is placed at the orifice of the catalytic cavity and coordinates a zinc ion, referred to as "a distal zinc ion", with His(131) . When the wild-type enzymes C122A (Cys(122)-->Ala), C124A (Cys(124)-->Ala), C132A (Cys(132)-->Ala) and C223A (Cys(223)-->Ala) were oxidized by hydrogen peroxide, the levels of activity were decreased. Two cysteines were titrated with 5,5' dithiobis(2-nitrobenzoic acid) in the wild-type enzyme, while on the other hand, one cysteine was titrated in the mutant enzymes. When wild-type and mutant enzymes were reduced by 2-mercaptoethanol, the levels of activity were increased: four and three cysteines were titrated, respectively, suggesting that a disulfide bond was formed among Cys(122), Cys(124) and Cys(132) under oxidizing conditions. We analyzed the enzyme-bound zinc ion of these enzymes using inductively coupled plasma mass spectrometry with gel-filtration chromatography. The results for C223A showed that the number of proximal zinc ions correlated to the level of enzymatic activity. Furthermore, zinc-ion-free 2-mercaptoethanol increased the activity of the wild-type enzyme without a change in the total number of zinc ions, but C223A was not activated. These findings suggest that a distal zinc ion moved to the proximal binding site when a disulfide bond among Cys(122), Cys(124) and Cys(132) was reduced by reductants. Thus, in the catalytic functioning of the enzyme, the distal zinc ion does not directly contribute but serves rather as a reserve as the next proximal one that catalyzes the enzyme reaction. A redox change of the three cysteines in the active center accommodates the catch and release of the reserve distal zinc ion placed at the orifice of the catalytic cavity. PMID- 15747134 TI - Vanadium-dependent iodoperoxidases in Laminaria digitata, a novel biochemical function diverging from brown algal bromoperoxidases. AB - The brown alga Laminaria digitata features a distinct vanadium-dependent iodoperoxidase (vIPO) activity, which has been purified to electrophoretic homogeneity. Steady-state analyses at pH 6.2 are reported for vIPO (K (m) (I-) = 2.5 mM; k (cat) (I-) = 462 s(-1)) and for the previously characterised vanadium dependent bromoperoxidase in L. digitata (K (m) (I-) =18.1 mM; k (cat) (I-) = 38 s(-1)). Although the vIPO enzyme specifically oxidises iodide, competition experiments with halides indicate that bromide is a competitive inhibitor with respect to the fixation of iodide. A full-length complementary ANA (cDNA) was cloned and shown to be actively transcribed in L. digitata and to encode the vIPO enzyme. Mass spectrometry analyses of tryptic digests of vIPO indicated the presence of at least two very similar proteins, in agreement with Southern analyses showing that vIPOs are encoded by a multigenic family in L. digitata. Phylogenetic analyses indicated that vIPO shares a close common ancestor with brown algal vanadium-dependent bromoperoxidases. Based on a three-dimensional structure model of the vIPO active site and on comparisons with those of other vanadium-dependent haloperoxidases, we propose a hypothesis to explain the evolution of strict specificity for iodide in L. digitata vIPO. PMID- 15747136 TI - Diffusion-weighted imaging-guided resection of intracerebral lesions involving the optic radiation. AB - In this paper we report our experience with diffusion-weighted imaging (DWI) for optic radiation (OR) visualization during resection of tumors. We hypothesize that intraoperative OR visualization helps to maintain patients' visual fields. DWI studies were performed together with T1-weighted postcontrast magnetic resonance imaging (MRI) in four patients with lesions in or adjacent to the OR (glioblastoma, oligo-astrocytoma, cavernoma, and metastasis; n = 1 each). The OR was identified from one of six DWI data acquisitions, segmented and reconstructed three-dimensionally. The image data were neuronavigationally transferred into the operative field, and provided the neurosurgeon with information on lesion site and adjacent OR localization. Preoperative and postoperative neuroophthalmological testing included, among others, perimetry to define the value of diffusion-weighted image guidance during OR lesion resection. Three lesions were removed completely. In one case, low-grade tumor parts infiltrating the OR were intentionally left. No persistent visual field deficits were induced. In one patient, a transient homonymous hemianopia attributable to postoperative swelling completely resolved under steroid medication. The authors conclude that intraoperative OR visualization, realized by neuronavigationally displayed DWI data, might prove to be helpful to maintain patients' visual fields. PMID- 15747137 TI - Cholesterol granulomas in antrochoanal polyps: a clinicopathologic study. AB - The purpose of this study was to investigate antrochoanal polyps with cholesterol granuloma (CG), which is a granulomatous reaction to cholesterol crystals that has been precipitated in the tissue. It is usually associated with chronic middle ear disease, common in the mastoid air cells, less common in the orbit and rarely found in the paranasal sinuses. The aim of this study was to analyze the etiology and pathological findings of antrochoanal polyps associated with cholesterol granuloma. This is a retrospective study of five cases of antrochoanal polyp with cholesterol granuloma, (four males and one female between 15 and 77 years of age) who presented with nasal obstruction, rhinorhea and snoring. The cases were clinically and histologically reviewed. Four patients were treated endoscopically and one by intranasal polypectomy without endoscope. There was no recurrence during the follow-up between 24 to 36 months (mean 31.2 months). Five uncommon cases with antrochoanal polyp with cholesterol granuloma are presented. The cholesterol granulomas consist of fibrous granulation tissue containing cholesterol crystals with surrounding foreign body giant cells. The pathogenesis of antrochoanal polyp with cholesterol granuloma is unclear, and further investigations are needed. PMID- 15747135 TI - Modulation of zinc- and cobalt-binding affinities through changes in the stability of the zinc ribbon protein L36. AB - Cysteine-rich Zn(II)-binding sites in proteins serve two distinct functions: to template or stabilize specific protein folds, and to facilitate chemical reactions such as alkyl transfers. We are interested how the protein environment controls metal site properties, specifically, how naturally occurring tetrahedral Zn(II) sites are affected by the surrounding protein. We have studied the Co(II)- and Zn(II)-binding of a series of derivatives of L36, a small zinc ribbon protein containing a (Cys)(3)His metal coordination site. UV-vis spectroscopy was used to monitor metal binding by peptides at pH 6.0. For all derivatives, the following trends were observed: (1) Zn(II) binds tighter than Co(II), with an average K (A) (Zn) /K (A) (Co) of 2.8(+/-2.0)x10(3); (2) mutation of the metal-binding ligand His32 to Cys decreases the affinity of L36 derivatives for both metals; (3) a Tyr24 to Trp mutation in the beta-sheet hydrophobic cluster increases K (A) (Zn) and K (A) (Co) ; (4) mutation in the beta-hairpin turn, His20 to Asn generating an Asn-Gly turn, also increases K (A) (Zn) and K (A) (Co) ; (5) the combination of His20 to Asn and Tyr24 to Trp mutations also increases K (A) (Zn) and K (A) (Co) , but the increments versus C(3)H are less than those of the single mutations. Furthermore, circular dichroism, size-exclusion chromatography, and 1D and 2D (1)H NMR experiments show that the mutations do not change the overall fold or association state of the proteins. L36, displaying Co(II)- and Zn(II) binding sensitivity to various sequence mutations without undergoing a change in protein structure, can therefore serve as a useful model system for future structure/reactivity studies. PMID- 15747138 TI - Long-term follow-up in patients with Pendred syndrome: vestibular, auditory and other phenotypes. AB - Fourteen patients with a Pendred syndrome gene (Pds) mutation and three patients without the mutation were studied to evaluate long-term vestibular and auditory manifestations among patients with bilateral enlarged vestibular aqueducts (EVA). Charts were reviewed retrospectively for age, gender, otological history, presence or absence of vertigo, results of magnetic resonance imaging, relevant gene mutations and perchlorate discharge test. A missense mutation, His 723 Arg (H723R), was identified in the homozygous state in six patients and in the heterozygous state in seven patients. Another missense mutation, Tyr 410 Met (T410 M), was identified in the heterozygous state in one patient. Patients with vertigo tended to have hearing fluctuation, compared with the patients without vertigo. Patients homozygous for H723R tended to have more episodes of vertigo than the heterozygous individuals. Only one patient who was homozygous for H723R had goiter. The long-term outcome of hearing level in patients with the H723R mutation was significantly better compared with published results for patients with a Pds mutation, but not for those with the H723R mutation. Whether environmental factors or a subtype of the Pds mutation H723R are related to the long-term outcome for these patients must be clarified. PMID- 15747139 TI - Occupations associated with bullae on chest radiographs in Japanese middle-aged men. AB - OBJECTIVES: To examine an association between occupation and bullae seen on chest radiographs. METHODS: We performed a cross-sectional study of 27,361 men in their 50s who consulted our institute for health screening. Logistic models that independently incorporated occupation, age and smoking as variables in a stepwise fashion were selected. RESULTS: Age and smoking significantly increased the odds ratio of the risk for developing bullae. For 19 occupations, the transport and the wholesale and retail trade industries were associated with significantly increased risk (odds ratios 1.6 and 1.5, respectively). In the transport industry, warehousing in particular showed increased risk by approximately 2.7 fold (95% CI 1.5-4.9). Road passenger transport, road freight transport, and retail were also associated with increased risk [odds ratios (5% CI) 1.3 (1.2 1.6), 1.4 (1.2-1.6) and 1.3 (1.04-1.5), respectively]. CONCLUSIONS: The finding of the present study suggests that independently of age or smoking, the prevalence of a bulla is associated with the warehouse, road transport, and retail industries in which workers may be remarkably exposed to exhaust gas. PMID- 15747140 TI - [Postoperative cognitive dysfunction]. AB - Postoperative cognitive dysfunction is a severe and life-threatening complication after an operation. The mobilisation of the patient is difficult and, therefore, the stay of the patient in the hospital is extended and the resulting immobilisation is associated with further complications (e.g. decubitus, pneumonia, or thrombosis). The genesis of the postoperative cognitive dysfunction has different causes and the highest risk factors are the age of the patient and the character and the duration of the operation. Preexisting diseases, like diabetes mellitus, heart failure, depression or alcohol excess are high risk factors as well as the use of anticholinergic drugs. A specific therapy for this cognitive dysfunction is unknown and, therefore, preventive measures should be used in patients with a preexisting high risk. PMID- 15747141 TI - [Locoregional anesthesia and coagulation inhibitors. Recommendations of the Task Force on Perioperative Coagulation of the Austrian Society for Anesthesiology and Intensive Care Medicine]. AB - More efficacious anticoagulant and antiplatelet agents have been introduced in vascular medicine and in the prevention of perioperative venous thromboembolisms. Patient management should be guided by familiarity with the pharmacology of coagulation-altering drugs and by consensus statements. The present paper reviews recommendations from the Austrian Task Force for Perioperative Coagulation which are based on thorough evaluation of the available pharmacological information and case reports. The consensus statement focuses on neuraxial and peripheral techniques and is designed to encourage safe and quality patient care. PMID- 15747142 TI - Improved correlation of histological data with DCE MRI parameter maps by 3D reconstruction, reslicing and parameterization of the histological images. AB - Due to poor correlation of slice thickness and orientation, verification of radiological methods with histology is difficult. Thus, a procedure for three dimensional reconstruction, reslicing and parameterization of histological data was developed, enabling a proper correlation with radiological data. Two different subcutaneous tumors were examined by MR microangiography and DCE-MRI, the latter being post-processed using a pharmacokinetic two-compartment model. Subsequently, tumors were serially sectioned and vessels stained with immunofluorescence markers. A ray-tracing algorithm performed three-dimensional visualization of the histological data, allowing virtually reslicing to thicker sections analogous to MRI slice geometry. Thick slices were processed as parameter maps color coding the marker density in the depth of the slice. Histological 3D reconstructions displayed the diffuse angioarchitecture of malignant tumors. Resliced histological images enabled specification of high enhancing areas seen on MR microangiography as large single vessels or vessel assemblies. In orthogonally reconstructed histological slices, single vessels were delineated. ROI analysis showed significant correlation between histological parameter maps of vessel density and MR parameter maps (r=0.83, P=0.05). The 3D approach to histology improves correlation of histological and radiological data due to proper matching of slice geometry. This method can be used with any histological stain, thus enabling a multivariable correlation of non-invasive data and histology. PMID- 15747143 TI - Enhanced fixation reveals the apical cortical fringe of actin filaments as a consistent feature of the pollen tube. AB - The actin cytoskeleton plays a crucial role in the growth and polarity of the pollen tube. Due to inconsistencies in the conventional preservation methods, we lack a unified view of the organization of actin microfilaments, especially in the apical domain, where tip growth occurs. In an attempt to improve fixation methods, we have developed a rapid freeze-whole mount procedure, in which growing pollen tubes (primarily lily) are frozen in liquid propane at -180 degrees C, substituted at -80 degrees C in acetone containing glutaraldehyde, rehydrated, quenched with sodium borohydride, and probed with antibodies. Confocal microscopy reveals a distinct organization of actin in the apical domain that consists of a dense cortical fringe or collar of microfilaments starting about 1-5 microm behind the extreme apex and extending basally for an additional 5-10 microm. In the shank of the pollen tube, basal to the fringe, actin forms abundant longitudinal filaments that are evenly dispersed throughout the cytoplasm. We have also developed an improved ambient-temperature chemical fixation procedure, modified from a protocol based on simultaneous fixation and phalloidin staining. We removed EGTA, elevated the pH to 9, and augmented the fixative with ethylene glycol bis[sulfosuccinimidylsuccinate] (sulfo-EGS). Notably, this protocol preserves the actin cytoskeleton in a pattern similar to that produced by cryofixation. These procedures provide a reproducible way to preserve the actin cytoskeleton; employing them, we find that a cortical fringe in the apex and finely dispersed longitudinal filaments in the shank are consistent features of the actin cytoskeleton. PMID- 15747144 TI - Suppression of heterotrimeric G-protein beta-subunit affects anther shape, pollen development and inflorescence architecture in tobacco. AB - The role of the heterotrimeric G-protein beta-subunit in plant development was studied in transgenic tobacco (Nicotiana tabacum L.) plants with reduced beta subunit levels due to the antisense expression of the beta-subunit mRNA. The antisense plants had aberrant anther shape and produced non-germinating pollen. The anthers were sporadically transformed to petals, whereas other floral organs were not affected. The pollen grains were smaller than the wild-type pollen and had abnormal cell walls. The architecture of mature antisense plants was altered. The plants had long branched panicles and short stems. These data suggest that the beta-subunit of the plant heterotrimeric G-proteins is involved in the regulation of the reproductive phase of the tobacco life cycle, particularly in stamen development and pollen maturation. PMID- 15747145 TI - Xylem parenchyma cells deliver the H2O2 necessary for lignification in differentiating xylem vessels. AB - Lignification in Zinnia elegans L. stems is characterized by a burst in the production of H(2)O(2), the apparent fate of which is to be used by xylem peroxidases for the polymerization of p-hydroxycinnamyl alcohols into lignins. A search for the sites of H(2)O(2) production in the differentiating xylem of Z. elegans stems by the simultaneous use of optical (bright field, polarized light and epi-polarization) and electron-microscope tools revealed that H(2)O(2) is produced on the outer-face of the plasma membrane of both differentiating (living) thin-walled xylem cells and particular (non-lignifying) xylem parenchyma cells. From the production sites it diffuses to the differentiating (secondary cell wall-forming) and differentiated lignifying xylem vessels. H(2)O(2) diffusion occurs mainly through the continuous cell wall space. Both the experimental data and the theoretical calculations suggest that H(2)O(2 )diffusion from the sites of production might not limit the rate of xylem cell wall lignification. It can be concluded that H(2)O(2) is produced at the plasma membrane in differentiating (living) thin-walled xylem cells and xylem parenchyma cells associated to xylem vessels, and that it diffuses to adjacent secondary lignifying xylem vessels. The results strongly indicate that non-lignifying xylem parenchyma cells are the source of the H(2)O(2) necessary for the polymerization of cinnamyl alcohols in the secondary cell wall of lignifying xylem vessels. PMID- 15747147 TI - [Elastic-stable-intramedullary nailing for forearm fractures in children using TENS]. PMID- 15747146 TI - [Accidental thermal injury to the hand]. AB - Hands are most frequently involved in severe burns. The treatment of the burned hand is one of the most challenging problems in plastic surgery. The success of therapy is determined by the severity of the initial injury, the first aid provided, and the correct balance between mobilization, splinting and plastic surgery. In this respect, it is vital to have a highly qualified interdisciplinary team approach consisting of plastic surgeons, well-trained physical therapists, psychologists and a highly motivated patient. PMID- 15747148 TI - [Bavarian mammography screening program]. AB - In Bavaria since the 1st April 2003 we have been conducting a high quality mammography-screening carried out in individual practises (BMS). We have used the European and the S 3 guidelines. The best diagnosis is an early diagnosis of the breast carcinoma to save human life. Because of this and the high mortality rate due to this disease it is essential to have a mammogram screening program. There is no single one ideal way of constructing a screening program, it is always based on compromise within the particular health care-systems. Arising problems cannot be avoided, it is only possible when all parties work closely together that the BMS works properly. PMID- 15747149 TI - [3D ultrasound (3D US) in the diagnosis of focal breast lesions]. AB - BACKGROUND: It was to analyse whether 3D breast US is able to enhance the diagnosis of focal breast lesions. MATERIAL AND METHODS: 60 patients were examined with 2D- and 3D US (GE logiq 9, 14 MHz). The solid lesions were analyzed by using the BIRADS classification. As standard of reference, histopathologic results were available in all cases. RESULTS: 38 malignant and 22 benign lesions were analyzed. With 2D US the sensitivity/specificity was 92/81%, with 3D US 97/72%, and by combination of 2D and 3D 97/81%. Characteristics of benign solid masses included a round or oval shape as well as clearly defined and sharply demarcated margins. CONCLUSIONS: 3D US allows to demonstrate breast masses in multiple planes. Distorsion of the surrounding soft tissues is an important sign indicative of malignancy. PMID- 15747150 TI - [The quality of digital mammograms. Development and use of phantoms for optimal safety]. AB - Digital imaging in mammography is becoming more and more accepted using both computed (CR) and direct radiography (DR). These techniques will soon be used in screening programs. Therefore, quality assurance for this technique is indispensable. The relevance of the current regulations, such as EPOC and the German QS-RL was investigated. For the investigation, a breast phantom and appropriate software were developed. Both were tested using digital mammography systems from six manufactures. Quality assurance parameters (such as contrast to noise ratio and contrast resolution) were calculated from these data sets. The results should be considered in future standards for mammography (IEC respectively DIN). In addition, this type of test procedure is time saving and enables a reduction in test devices, i. e. in costs. PMID- 15747153 TI - Hybrid SPECT/CT: a new era for SPECT imaging? PMID- 15747151 TI - Resistance to cyclic loading of teeth restored with posts. AB - This in vitro study evaluated the effect of presence of post, presence of core, and of shape, type, and surface treatment of posts on resistance to cyclic loading of crowned human teeth. For all teeth, crowns designed without ferrule were cast in sterling silver and luted with resin cement (Panavia F). Each tooth underwent cyclic loading of 600 N at two loads per second until failure. Teeth that had only been crowned showed significantly higher resistance to cyclic loading than teeth with cores or with post and cores. No significant differences were found between teeth restored with cores only or with post and cores, irrespective of surface-treatment of the posts. Teeth restored with parallel sided cast post (ParaPost XP) and cores showed significantly higher resistance to cyclic loading than teeth with either tapered cast posts or untreated prefabricated posts of titanium alloy (ParaPost XH) or glass fiber composite (ParaPost Fiber White). No significant difference was found between teeth restored with parallel-sided cast post and cores and teeth restored with untreated prefabricated posts of zirconia (Cerapost). Surface treatment of posts significantly increased the resistance to cyclic loading compared with untreated posts. When posts are used, surface treatment is recommended. PMID- 15747154 TI - The value of [18F]FDG-PET in the diagnosis of large-vessel vasculitis and the assessment of activity and extent of disease. AB - PURPOSE: This study was performed to investigate the value of( 18)F fluorodeoxyglucose positron emission tomography ([(18)F]FDG-PET) in the diagnosis of large-vessel vasculitis and the assessment of activity and extent of disease. METHODS: Twenty-six consecutive patients (21 females, 5 males; median age - years, range 17-86 years) with giant cell arteritis or Takayasu's arteritis were examined with [(18)F]FDG-PET. Follow-up scans were performed in four patients. Twenty-six age- and gender-matched controls (21 females, 5 males; median age 71 years, range 17-86 years) were included. The severity of large-vessel [(18)F]FDG uptake was visually graded using a four-point scale. C-reactive protein (CRP) and the erythrocyte sedimentation rate (ESR) were measured and correlated with [(18)F]FDG-PET results by logistic regression. RESULTS: [(18)F]FDG-PET revealed pathological findings in 18 of 26 patients. Three scans were categorised as grade I, 12 as grade II and 3 as grade III arteritis. Visual grade was significantly correlated with both CRP and ESR levels (p=0.002 and 0.007 respectively; grade I: CRP 4.0 mg/l, ESR 6 mm/h; grade II: CRP 37 mg/l, ESR 46 mm/h; grade III: CRP 172 mg/l, ESR 90 mm/h). Overall sensitivity was 60% (95% CI 40.6-77.3%), specificity 99.8% (95% CI 89.1-100%), positive predictive value 99.7% (95% CI 77-100%), negative predictive value 67.9% (95% CI 49.8-80.9%) and accuracy 78.6% (95% CI 65.6-88.4%). In patients presenting with a CRP <12 mg/l or an ESR <12 mm/h, logistic regression revealed a sensitivity of less than 50%. In patients with high CRP/ESR levels, sensitivity was 95.5%/80.7%. CONCLUSION: [(18)F]FDG-PET is highly effective in assessing the activity and the extent of large-vessel vasculitis. Visual grading was validated as representing the severity of inflammation. Its use is simple and provides high specificity, while high sensitivity is achieved by scanning in the state of active inflammation. PMID- 15747155 TI - Limited pelvic lymphadenectomy using the sentinel lymph node procedure in patients with localised prostate carcinoma: a pilot study. AB - PURPOSE: The purpose of this study was to determine the potential role of the sentinel lymph node (SLN) procedure in limited lymph node dissection in patients with apparently localised prostate carcinoma. METHODS: In 27 patients with organ confined prostate cancer, a single injection of 0.3 ml/30 MBq( 99m)Tc-rhenium sulphur colloid was injected transrectally into the peripheral zone of each lobe of the prostate (total 0.6 ml/60 MBq) under ultrasound guidance. Two hours after injection, scintigraphy was performed. The first step in surgery was the detection and dissection of lymph nodes identified as SLNs. Then, standard lymphadenectomy was performed, consisting in a limited dissection that included all lymph nodes from the obturator fossa and along the external iliac vein. Lymphatic tissue along the hypogastric artery was not systematically removed, except in the presence of SLNs. RESULTS: Mean patient age was 66 years (48-77); the mean serum prostate-specific antigen value was 10.6 ng/ml. In a high proportion of patients (21/27, 77.8%) an SLN was located along the initial centimetres of the hypogastric artery. The second most frequent site of SLNs was in the obturator fossa (11/27 patients, 40.7%), followed by the external iliac area (5/27 patients, 18.5%). Four patients had lymph node metastases, all in SLNs: two in the hypogastric area and two in the obturator fossa. CONCLUSION: The SLN procedure revealed the individual variability in the lymphatic drainage of the prostate. The main site of SLNs was the hypogastric area, and two of the four metastatic nodes were located at this site. A limited standard pelvic lymphadenectomy, excluding the hypogastric lymph nodes, would have missed half of the lymph node metastases in this study. A radionuclide SLN procedure could assist in the correct staging of patients with early prostate cancer, especially when performing limited lymphadenectomy. PMID- 15747152 TI - Brain glucose metabolism in the early and specific diagnosis of Alzheimer's disease. FDG-PET studies in MCI and AD. AB - The demographics of aging suggest a great need for the early diagnosis of dementia and the development of preventive strategies. Neuropathology and structural MRI studies have pointed to the medial temporal lobe (MTL) as the brain region earliest affected in Alzheimer's disease (AD). MRI findings provide strong evidence that in mild cognitive impairments (MCI), AD-related volume losses can be reproducibly detected in the hippocampus, the entorhinal cortex (EC) and, to a lesser extent, the parahippocampal gyrus; they also indicate that lateral temporal lobe changes are becoming increasingly useful in predicting the transition to dementia. Fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) imaging has revealed glucose metabolic reductions in the parieto temporal, frontal and posterior cingulate cortices to be the hallmark of AD. Overall, the pattern of cortical metabolic changes has been useful for the prediction of future AD as well as in distinguishing AD from other neurodegenerative diseases. FDG-PET on average achieves 90% sensitivity in identifying AD, although specificity in differentiating AD from other dementias is lower. Moreover, recent MRI-guided FDG-PET studies have shown that MTL hypometabolism is the most specific and sensitive measure for the identification of MCI, while the utility of cortical deficits is controversial. This review highlights cross-sectional, prediction and longitudinal FDG-PET studies and attempts to put into perspective the value of FDG-PET in diagnosing AD-like changes, particularly at an early stage, and in providing diagnostic specificity. The examination of MTL structures, which has so far been exclusive to MRI protocols, is then examined as a possible strategy to improve diagnostic specificity. All told, there is considerable promise that early and specific diagnosis is feasible through a combination of imaging modalities. PMID- 15747156 TI - Voxel-based statistical analysis of cerebral glucose metabolism in the rat cortical deafness model by 3D reconstruction of brain from autoradiographic images. AB - PURPOSE: Animal models of cortical deafness are essential for investigation of the cerebral glucose metabolism in congenital or prelingual deafness. Autoradiographic imaging is mainly used to assess the cerebral glucose metabolism in rodents. In this study, procedures for the 3D voxel-based statistical analysis of autoradiographic data were established to enable investigations of the within modal and cross-modal plasticity through entire areas of the brain of sensory deprived animals without lumping together heterogeneous subregions within each brain structure into a large region of interest. METHODS: Thirteen 2-[1-(14)C] deoxy-D: -glucose autoradiographic images were acquired from six deaf and seven age-matched normal rats (age 6-10 weeks). The deafness was induced by surgical ablation. For the 3D voxel-based statistical analysis, brain slices were extracted semiautomatically from the autoradiographic images, which contained the coronal sections of the brain, and were stacked into 3D volume data. Using principal axes matching and mutual information maximization algorithms, the adjacent coronal sections were co-registered using a rigid body transformation, and all sections were realigned to the first section. A study-specific template was composed and the realigned images were spatially normalized onto the template. Following count normalization, voxel-wise t tests were performed to reveal the areas with significant differences in cerebral glucose metabolism between the deaf and the control rats. RESULTS: Continuous and clear edges were detected in each image after registration between the coronal sections, and the internal and external landmarks extracted from the spatially normalized images were well matched, demonstrating the reliability of the spatial processing procedures. Voxel-wise t tests showed that the glucose metabolism in the bilateral auditory cortices of the deaf rats was significantly (P<0.001) lower than that in the controls. There was no significantly reduced metabolism in any other area, and no area showed a significant increase in metabolism in the deaf rats with the same threshold, demonstrating the high localization accuracy and specificity of the method developed in this study. CONCLUSION: This study established new procedures for the 3D reconstruction and voxel-based analysis of autoradiographic data which will be useful for examining the cerebral glucose metabolism in a rat cortical deafness model. PMID- 15747157 TI - Extension of myocardial necrosis differently affects MIBG retention in heart failure caused by ischaemic heart disease or by dilated cardiomyopathy. AB - PURPOSE: This study aims to investigate the relationship between cardiac sympathetic nervous function (CSNF) and myocardial perfusion/function in patients with heart failure (HF) due to dilated cardiomyopathy (DCM) or ischaemic heart disease (CAD). METHODS: Twenty patients (10 DCM, 10 CAD, 17 males, age 69+/-5 years) with NYHA class IIIb HF were studied. CSNF was evaluated by early/delayed (123)I-metaiodobenzylguanidine (MIBG) uptake and regional washout (WO). Myocardial perfusion and function were evaluated by (99m)Tc-tetrofosmin gated single-photon emission tomography (G-SPECT) using a 20-segment model for 400 segments. In each segment, regional MIBG WO was computed as (count density in early images-count density in delayed images/count density in early images)x100. RESULTS: DCM and CAD showed similar summed rest perfusion score (6.7+/-5 vs 9.5+/ 5, p=NS) and mean ejection fraction values (29+/-7% vs 30+/-9%, p=NS). By contrast, the summed thickening score was higher in DCM than in CAD patients (26+/-7 vs 17+/-6, p<0.05). QGS analysis identified akinesis/dyskinesis in 129/137 (94%) severely hypoperfused segments which were considered as damaged. According to the underlying aetiology of HF, marked differences in regional MIBG WO were observed. In fact, within the CAD group, regional MIBG WO was lower in reference than in damaged segments (38+/-21% vs 46+/-19%, p<0.05). By contrast, in DCM patients, regional MIBG WO was faster in reference than in damaged segments (49+/-18% vs 41+/-30%, p<0.05). When the two groups were directly compared, regional MIBG WO from damaged areas was similar irrespective of the underlying disease, while it was faster in DCM than in CAD patients from reference segments. CONCLUSION: These data confirm the hypothesis that the presence of myocardial necrosis in HF due to CAD and the consequent loss of neuronal endings cause alterations in regional MIBG WO different from those observed in DCM. PMID- 15747158 TI - Determination of cell concentration in a plant cell suspension using a fluorescence microplate reader. AB - Microscopic counting of plant cells is a very tedious and time-consuming process and is therefore seldom used to evaluate plant cell number on a routine basis. This study describes a fast and simple method to evaluate cell concentration in a plant cell suspension using a fluorescence microplate reader. Eschscholtzia californica cells were fixed in a mix of methanol and acetic acid (3:1) and stained with a fluorescent DNA binding dye (Hoechst 33258). Readings were done in a fluorescence microplate reader at 360/465 nm. Specific binding of the dye to double-stranded DNA was significantly favored over unspecific binding when 1.0 M Tris buffer at pH 7.5 containing 1.0 M NaCl and 75 microg ml(-1) of Hoechst 33258 was used. Fluorescence readings must be done between 4 min and 12 min following the addition of the staining solution to the sample. The microplate counting method provides a convenient, rapid and sensitive procedure for determining the cell concentration in plant cell suspensions. The assay has a linear detection range from 0.2 x 10(6) cells to 10.0 x 10(6) cells per milliliter (actual concentration in the tested cell suspension). The time needed to perform the microplate counting was 10% of that needed for the microscopic enumeration. However, this microplate counting method can only be used on genetically stable cell lines and on asynchronous cell suspensions. PMID- 15747159 TI - Vitreoscilla hemoglobin overexpression increases submergence tolerance in cabbage. AB - Agrobacterium tumefaciens was used to deliver the vhb gene into cabbage (Brassica oleracea var. Cabitata) cv. Xiaguang's parent line, 103. Using hypocotyls and cotyledon petioles as explants for infection, we obtained a transformation efficiency of 3-5% based on the number of transgenic shoots produced from the number of explants used for infection. Molecular analysis indicated that the vhb gene was stably integrated into the cabbage genome and that the vhb gene was expressed at the RNA level. Characterization of the vhb over-expressing transgenic plants revealed that transgenic seeds germinated faster than the wildtype controls. More importantly, the transgenic plants showed tolerance to a prolonged submergence treatment, suggesting that the vhb gene may provide an excellent tool for creation of submergence/flooding-tolerant cultivars of agriculturally important crops. PMID- 15747160 TI - Distribution of ferritin in the rat hippocampus after kainate-induced neuronal injury. AB - A gradual increase in iron occurs in the lesioned hippocampus after neuronal injury induced by the excitotoxin kainate, and the present study was carried out to investigate whether this increase in iron might be associated with changes in expression of the iron binding protein, ferritin. An increase in ferritin immunoreactivity was observed in glial cells of the hippocampus, as early as three days after intracerebroventricular injections of kainate. The number of ferritin positive cells peaked four weeks after the kainate injection, and decreased eight and twelve weeks after injection. They were found to be mostly microglia and oligodendrocytes by double immunofluorescence labeling with glial markers. A number of ferritin-labeled endothelial cells were also observed via electron microscopy. The decline in ferritin immunoreactivity four weeks after the injection of kainate is accompanied by an increase in the number of ferric and ferrous iron positive cells in the lesioned tissue. A substantial non-overlap between ferritin and iron-containing cells was observed. In particular, spherical ferric or ferrous iron-laden cells in the degenerating hippocampus were unlabeled for ferritin for long time periods after the kainate injection. An increase in iron, together with a reduced expression of iron binding proteins such as ferritin at long time intervals after kainate lesions, could result in a relative decrease in ferritin-induced ferroxidase activity and the presence of some of the iron in the ferrous form. It is postulated that this may contribute to chronic neuronal injury, following acute kainate-induced neurodegeneration. PMID- 15747161 TI - First NIH/Office of Rare Diseases Conference on Cystinosis: past, present, and future. PMID- 15747162 TI - Laparoscopic renal biopsy in obese children. AB - Percutaneous kidney biopsy is routinely used to obtain renal tissue for histological examination. It is usually successful and has very few contraindications. We describe two children with clinical obesity in whom the percutaneous approach failed to yield renal tissue for histology. They underwent successful laparoscopic kidney biopsies that yielded adequate renal tissue for diagnosis, although the first attempt in one patient yielded renal medulla necessitating a repeat biopsy. We recommend that laparoscopic kidney biopsies should be considered in obese children when percutaneous kidney biopsies are considered impossible. PMID- 15747163 TI - Central nervous system vasculitis secondary to parvovirus B19 infection in a pediatric renal transplant patient. AB - Central nervous system (CNS) vasculitis secondary to chronic parvovirus B19 (B19) infection presenting with recurrent neurological findings is a very rare disorder during childhood. Here we report a 12-year-old boy with a renal transplant who had chronic B19 infection with skin eruptions and recurrent episodes of encephalopathy with focal neurological deficits. B19 DNA was detected in blood, bone marrow, and skin biopsy specimens. Repeat cranial magnetic resonance (MR) imaging during each episode of encephalopathy showed variable focal findings, and MR angiography revealed vasculitic changes with narrowing of the cerebral arteries. We hypothesized that the CNS vasculitis might be associated with the chronic B19 infection. At the time of his fourth presentation with the same clinical findings, we administered intravenous immunoglobulin (IVIG) (1 g/kg per day, 2 consecutive days), which we continued for 6 months, at monthly intervals. IVIG therapy resulted in remission and has been effective not only for the clearance of B19, but also for the improvement of clinical and radiological findings of CNS vasculitis. We suggest that chronic B19 infection should be considered in immunocompromised patients with suspected CNS vasculitis. IVIG should be considered as a part of the treatment. PMID- 15747164 TI - Quantitative detection of serum HBV DNA levels employing a new S gene based cPCR assay. AB - Hepatitis B virus (HBV) infection is a major public health problem and a leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma. Worldwide, there are about 350 million carriers of this pathogen and India bears the second highest carrier pool in the world. Early diagnosis and measurement of viral load in hepatitis B patients is very helpful for the better management of this disease. The existing methods for viral quantification are either cumbersome or expensive. Since viral replication correlate well with HBV DNA levels a new sensitive, reliable and cost effective competitive PCR assay has been developed for quantifying the viral load in the serum of hepatitis B patients. The S gene based cPCR assay was able to detect as low as 100 genome equivalent/ml of HBV DNA from human serum and was applied to determine viral load among inactive and chronic hepatitis B carriers demonstrating the usefulness of the developed test. PMID- 15747165 TI - Cetacean morbilliviruses are phylogenetically divergent. AB - We performed a phylogenetic comparison of porpoise morbillivirus (PMV) and dolphin morbillivirus (DMV) isolates from porpoises and dolphins respectively according to criteria adopted by the World Health Organization for the phylogenetic comparison of measles viruses. PMV and DMV were more divergent than the most distantly related measles virus strains, thus challenging the classification of PMV and DMV as two strains of a single species, cetacean morbillivirus. PMID- 15747166 TI - Paternal uniparental disomy of chromosome 14 and unique exchange of chromosome 7 in cases of spontaneous abortion. AB - To investigate the involvement of uniparental disomies (UPDs) in spontaneous abortion, the polymorphic patterns of microsatellites on each chromosome were analyzed in 164 cases of abortion. Eighty-three of the 164 cases had chromosomal abnormalities. In 79 of the remaining 81 cases with normal karyotypes, the microsatellite analysis revealed that biparental patterns were present in the informative microsatellites in all chromosomes. In one of the remaining two cases, however, the polymorphic patterns of chromosome 14 appeared to be both of paternal origin. The patterns of the distal of the long arm were homozygous, and those of the remaining region were heterozygous. That is, this fetus had paternal UPD 14, originating from meiosis I nondisjunction. In the other case, the polymorphic patterns of the distal one third of the long arm of chromosome 7 were uniparental (maternal) in origin whereas those of the remaining region of this chromosome were biparental. These findings thus suggested that this chromosome might have originated from chromatid exchange between the long arms of paternal and maternal chromosome 7 at the first mitotic division. Microsatellite analysis, however, produced no evidence of duplication or deletion of any segments. The findings also suggest the possibility that some UPDs may cause spontaneous abortion. PMID- 15747167 TI - Pancreaticoduodenectomy for advanced gastric cancer. AB - BACKGROUND: Although pancreaticoduodenectomy has been rarely performed for gastric cancer because of frequent morbidity and mortality, some favorable results after this procedure have been reported recently. Our objective was to present our data that might aid in the selection of patients to undergo this procedure. METHODS: Between 1970 and 2001, 23 patients who had pancreaticoduodenectomy for gastric cancer with tumor invading the pancreatic head were identified, and they were the subjects of this study. Clinical, operative, and pathological data, and morbidity and mortality rates were collected and analyzed. Survival outcome was also calculated and analyzed. RESULTS: Five patients underwent this procedure for disease in the gastric remnant, 18 undergoing the procedure for primary tumors. Median operating time was 8 h (range, 6-13 h), and median blood loss was 1600 ml (range, 700-16,000 ml). Regarding extent of gastrectomy, all patients with primary cancer (n = 18) underwent a distal gastrectomy and patients with disease in the gastric remnant (n = 5) underwent a completion gastrectomy. Incurable factors, including paraaortic lymph node metastasis, positive lavage cytology, or peritoneal dissemination were found in 8 patients. The postoperative morbidity rate was 73.9%; however, operation-related death was zero. The overall 5-year survival rate was 34.3%. The 5-year survival rate of the 8 patients with incurable factors was 0%, while that of the 15 patients without incurable factors was 47.4%. CONCLUSION: If an R0 resection can be achieved by pancreaticoduodenectomy, this procedure should be performed for patients with tumor invading the pancreatic head. Patients with incurable factors should not be considered for pancreaticoduodenectomy. PMID- 15747168 TI - Japanese nationwide post-marketing survey of S-1 in patients with advanced gastric cancer. AB - BACKGROUND: It is likely that there are some discrepancies in the safety and efficacy results for anticancer agents between those shown in registration studies for approval and those shown in clinical practice after market release. The aim of this survey was to confirm the safety and efficacy of S-1 for advanced gastric cancer after market release. METHODS: After the approval of S-1 in 1999, all patients had to be registered with the manufacturer for a post-marketing survey, according to the government recommendation. All patients were monitored for safety and survival. The data for all registered patients were updated 1 year after each registration. RESULTS: During this survey, a total of 4177 patients with advanced gastric cancer were registered. The incidences of all adverse events and of grade 3 or worse adverse events in the 3808 patients evaluable for safety were 74.3% and 25.0%, respectively. In patients with lower creatinine clearance at baseline, the incidences of adverse reactions were higher for all grades combined, as well as for grades 3 or worse. There were 90 (2.4%) early deaths (within 30 days of the initiation of the treatment) and 5 (0.1%) deaths possibly related to the treatment. The median survival time and the 1-year survival rate for all patients evaluable for efficacy (n = 3801) were 8.3 months (95% confidence interval [CI], 8.0-8.6 months) and 33.3% (95% CI, 31.8-34.9%), respectively. CONCLUSION: This nationwide survey confirmed that the safety and efficacy profiles of S-1 were similar to those seen in the registration study. These results have proven the utility of this post-marketing survey in assessing the reproducibility of the safety and efficacy results obtained from prior clinical studies. PMID- 15747169 TI - Association between Helicobacter pylori seropositivity and NAD(P)H:quinone oxidoreductase 1 (NQO1) C609T polymorphism observed in outpatients and health checkup examinees. AB - BACKGROUND: Significant associations of Helicobacter pylori (H. pylori) seropositivity have been found with several host polymorphisms. This study investigated the associations of functional polymorphisms of the NQO1, GSTM1, and GSTT1 genes of detoxification enzymes, with the seropositivity, as well as with pepsinogen levels, as markers of gastric atrophy. METHODS: The subjects were 241 noncancer outpatients who had participated in an H. pylori eradication program (HPE) at Aichi Cancer Center Hospital, and 465 health checkup examinees in Nagoya (HCE). The NQO1 C609T, GSTM1, and GSTT1 polymorphisms were determined by triplex polymerase chain reaction with confronting two-pair primers (PCR-CTPP). RESULTS: The sex- and age-group-adjusted odds ratio (OR) of NQO1 C/C for H. pylori seropositivity relative to T/T was highly significant; OR, 1.92; 95% confidence interval (95% CI), 1.22-3.03. The ORs of the GSTM1 present type and GSTT1 present type for H. pylori seropositivity were not significant; OR, 0.87; 95% CI, 0.64 1.20 and OR, 1.14; 95% CI, 0.83-1.57, respectively. The association of the NQO1 C/C genotype with H. pylori seropositivity was observed only for never-smokers; OR, 2.25; 95% CI, 1.33-3.79. The genotypes of the NQO1, GSTM1, and GSTT1 genes were not associated with the development of atrophic gastritis among the H. pylori-seropositive subjects. CONCLUSION: This is the first study to report a significant association of the NQO1 C609T polymorphism with H. pylori seropositivity. The biological mechanism explaining the significant association with the seropositivity remains to be elucidated. PMID- 15747170 TI - Expression of endothelial nitric oxide synthase correlates with the angiogenic phenotype of and predicts poor prognosis in human gastric cancer. AB - BACKGROUND: Angiogenesis is a critical aspect of cancer biology and is subject to regulation by multiple molecular pathways. We evaluated the expression of nitric oxide synthase (NOS) III and its relationship with the angiogenic phenotype and expression of the transcription factor Sp1, as well as their effect on survival in patients with gastric cancer. METHODS: The NOS III expression level and tumor microvessel density (MVD) status were determined via immunohistochemistry, using archived tissue specimens from 86 resected gastric cancer cases; these findings were then correlated with Sp1 expression and clinicopathological features. RESULTS: NOS III protein expression was significantly higher in both primary tumors and lymph node metastases than in normal gastric mucosa. In primary tumors, NOS III expression correlated highly with Sp1 expression (P = 0.001) and MVD status (P = 0.001). Patients with strong Sp1 expression were more likely to have strong NOS III expression (15 times) and a high MVD (7 times) than were those with negative Sp1 expression. In univariate survival analyses, strong NOS III expression (P = 0.042), strong Sp1 expression (P = 0.007), and a high MVD (P = 0.036) were associated with worse survival. However, when patients' NOS III and Sp1 expression levels, MVD status, disease stage, completeness of resection, Lauren's classification, and age were entered in a Cox proportional hazards model, only strong NOS III (P = 0.019) and Sp1 expression (P = 0.029) and advanced disease stage (P = 0.006) were independently prognostic of poor survival. CONCLUSION: Our results indicated that the expression of NOS III, a potential downstream effector of Sp1, may play an important role in tumor angiogenesis and aggressiveness in gastric cancer. PMID- 15747171 TI - Progressive CT system technology and experience improve the perceived preoperative stage of gastric cancer. AB - BACKGROUND: Our goals were to measure the accuracy of specialist helical computed tomography (CT) in the preoperative staging of gastric cancer, to determine the relative benefit of progressive CT system technology, and to determine the magnitude of any learning curve in radiological interpretation. METHODS: One hundred patients (median age, 70 years; range 27-86 years; 68 male) underwent a preoperative CT (73 helical [hCT], 27 multislice [mCT]), performed by a single specialist radiologist, followed by surgery within 3 weeks. The strength of the agreement between the perceived CT stage and the histopathological stage was determined for each CT system and also for four serial cohorts of 25 patients, by the weighted Kappa statistic (Kw). RESULTS: The Kw values for T, N, M1 liver, and M1 peritoneal stage were 0.40, 0.18, 0.36, and 0.09 for hCT, compared with 0.57, 0.67, 0.66 (all P < 0.001), and 0.24 (P = 0.06) for mCT. Serial Kw for T and N stages improved from 0.26 and -0.14 in the first quartile of patients to 0.61 and 0.73 (P < 0.001) in the last quartile of patients. CONCLUSION: The role of CT in the preoperative staging of gastric cancer is becoming stronger as CT technology improves. PMID- 15747172 TI - A rare complication of D3 dissection for gastric carcinoma: chyloperitoneum. AB - BACKGROUND: Chyloperitoneum is the accumulation of lymphatic fluid in the peritoneal cavity. Although uncommon, it has been reported after retroperitoneal lymph node dissection. But the incidence of this complication after radical gastrectomy is unknown. In the present study, we analyzed our patients who underwent D3 dissection for gastric carcinoma and developed chyloperitoneum. METHODS: Between June 1999 and June 2002, a total of 134 patients with gastric cancer underwent radical lymph node dissection, performed according to the Japanese Research Society for Gastric Cancer guidelines, as the standard procedure for gastric cancer treatment. Of these patients, 34 underwent D3 lymphadenectomy, and chyloperitoneum was detected in 4 of them. RESULTS: There were three male patients and one female patient. All patients were in stage III according to the International Union Against Cancer (UICC)-TNM classification. In three patient, chyle leakage was noticed during the surgery, and surgical ligation of the duct was performed. Abdominal distension developed in one patient 7 days after the surgery, and chylous ascites was diagnosed. This patient was success-fully treated with fasting and total parenteral nutrition, within 2 weeks. CONCLUSION: The incidence of chyloperitoneum is not low, and may increase with more aggressive surgery. Surgeons should be aware of this complication after retroperitoneal lymph node dissection, and injured lymphatics must be controlled and ligated intraoperatively. PMID- 15747173 TI - Increasing body mass index in Japanese patients with gastric cancer. AB - We studied the body mass index (BMI) of 986 patients who underwent potentially curative gastrectomy for gastric cancer at the National Cancer Center Hospital, Tokyo, in 1971, 1981, 1991, and 2001. The median BMI increased from 20.8 kg/m(2) in 1971 to 22.6 kg/m(2) in 2001 (P, 0.01). The increase was significant in both early and advanced gastric cancers, and in males, but not in females. The proportion of overweight patients (BMI > or = 25.0 kg/m(2)) increased from 9.2% in 1971 to 24.0% in 2001. Obese patients (BMI > or = 30.0 kg/m(2)) were rare. In conclusion, surgeons at the National Cancer Center Hospital, Tokyo, are increasingly having to operate on fat patients, but obese patients are still uncommon compared to the West. PMID- 15747174 TI - Long-term follow up of patients with multiple gastric carcinoids associated with type A gastritis. AB - Little is known about the natural course of multiple gastric carcinoids associated with type A gastritis. Between 1993 and 2003, we enrolled eight patients, diagnosed as having multiple gastric carcinoids associated with type A gastritis, in a follow-up program without surgical resection. In these patients, endoscopy showed multiple small polyps on the gastric body, with nonantral atrophic gastritis. Histologically, biopsy specimens obtained from the polyps revealed carcinoid tumors. The serum gastrin level was found to be very high in all patients, and testing for anti-parietal cell antibody was positive in seven. The mean follow-up was 5.8 years (range, 1.5-10.8 years). The levels of serum gastrin increased in all patients, but, endoscopically, the carcinoid tumors did not change in size. Neither hepatic nor lymphatic metastasis was detected on abdominal computed tomography (CT). These patients were free of the development or metastasis of carcinoids, in spite of their continuous hypergastrinemia. It was concluded that multiple gastric carcinoids associated with type A gastritis may be indolent. PMID- 15747175 TI - Metastatic signet ring cell gastric carcinoma presenting as an infrarenal aortic aneurysm. AB - Metastases to liver, lungs, bone, and adrenal glands are common events in advanced gastric carcinoma. Occasionally, metastases to other parts of the body, such as the prostate gland [1] the gluteal muscle [2], or the cervix [3] are described. However, these are rare events in the natural history of the disease. We report an unusual case of a signet ring cell gastric carcinoma, initially presenting as an infrarenal aortic aneurysm. Following resection of the aneurysm, the spread of lymphangiosis carcinomatosa into the aortic wall and infiltration of signet ring cells into an adjacent lymph node were noted. The primary tumor, a signet ring cell gastric carcinoma, was detected by a subsequent esophago-gastro duodenoscopy. PMID- 15747176 TI - Neoadjuvant chemotherapy with S-1 and surgical resection for a mucinous gastric cancer with peritoneal dissemination. AB - We herein report the case of a patient with mucinous gastric carcinoma with peritoneal dissemination that disappeared after neoadjuvant chemotherapy with S-1 alone. The patient has survived for over 23 months after surgery, without recurrence. A 60-year old man was referred to our hospital because of an advanced gastric cancer, detected by upper gastrointestinal endoscopy at another hospital. Staging laparoscopy was performed on October 25, 2002, and revealed massive peritoneal dissemination. Two courses of neoadjuvant chemotherapy with S-1 were administered, at 120 mg/day for 28 days, as one course. Total gastrectomy, with D2 lymph node dissection, was performed on January 24, 2003. The peritoneal dissemination had macroscopically disappeared and the cytology of the peritoneal lavage fluid was class III. His final diagnosis was gastric carcinoma, MLU, type 3, T2(SS), P0, H0, M0, N3, CY0, stage IV. PMID- 15747177 TI - Phase development and structural characterization of calcium phosphate ceramics polyacrylic acid nanocomposites at room temperature in water-methanol mixtures. AB - Calcium phosphate ceramics (CPCs) were prepared via an in-situ formation in the presence of polyacrylic acid (PAA) polymer under water-methanol (WM) mixture at room temperature. The PAA polymer was employed as both structure-directing agent and crystallization retardant to manipulate the development of resulting CPCs nano-crystallites which are observed to show a core-shell configuration with a thin layer of PAA molecules. A resulting phase evolution map with respect to the developing phases of calcium-deficient hydroxyapatite (CDHA), beta-tricalcium phosphate (beta-TCP), and an intermediate amorphous calcium phoshate (ACP) that were structurally and spectroscopically identified, was constructed in terms of fractions of water-to-methanol proportions and concentration of PAA. It is found that for the solutions in both water-rich and methanol-rich regions, pure CDHA and beta-TCP instead of intermediate ACP phase can be developed irrespective of the concentration of PAA, respectively. For conditions in between, i.e., with methanol fractions of 15%-90%, ACP appeared only when the PAA fell in a limited concentration range. PMID- 15747178 TI - New fast method for determination of number of UHMWPE wear particles. AB - Ultra-high molecular weight polyethylene (UHMWPE) wear particles are the major cause of total joint replacement (TJR) failures because the wear particles, released from TJR's, cause bone loosening. To simplify the study of the relationship between numbers of particles at various locations around TJR's and extent of bone loosening at these locations, the authors of this work tried to develop a new method for easy and fast determination of number of wear particles. The method, called LSC (Light Scattering with Calibration spheres), is based on light scattering of a suspension of wear particles and calibration spheres, and yields relative numbers of particles. A modified LSC method, called LSCm, requires one additional experiment, a gravimetric analysis of a mixture of all studied samples, to determine absolute numbers of wear particles. LSC and LSCm methods are easy and fast, which make them suitable for processing and comparing high number of samples. PMID- 15747180 TI - Preparation and characterization of bioactive sol-gel-derived Na2Ca2Si3(O)9. AB - In this study, pure Na2Ca2Si3(O)9 was synthesized by a sol-gel method, and Na2Ca2Si3(O)9 cuboids and disks were prepared by uniaxial pressing and calcining at 700 degrees C. The porosity and mechanical strength of the Na2Ca2Si3(O)9 cuboids were measured, and the results showed that the Na2Ca2Si3(O)9 cuboids were porous with an average porosity of 44%, and the 3-point bending strength of the cuboids was 6.08 MPa. The in vitro bioactivity of Na2Ca2Si3(O)9 was carried out by soaking Na2Ca2Si3(O)9 disks in simulated body fluid (SBF). The results showed that hydroxyapatite (HA) formed on the surface of Na2Ca2Si3(O)9 samples after soaking for 1 day, which indicated good bioactivity of Na2Ca2Si3(O)9. PMID- 15747179 TI - Microwave accelerated synthesis of nanosized calcium deficient hydroxyapatite. AB - Rapid synthesis of calcium deficient hydroxyapatite (CDHA, Ca(10-x)(HPO4)x(PO4)(6 x)(OH)(2-x)) with Ca/P ratio 1.5 was done by precipitation using calcium nitrate tetra-hydrate and phosphoric acid and subsequently subjecting to microwave irradiation in a domestic microwave oven for 15 min. Transmission electron microscopy analysis shows needle like morphology of CDHA having length 16-39 nm and width 7-16 nm. The synthesized CDHA has the characteristic HPO4(2-) vibration band at 875 cm(-1) in Fourier transform infrared (FT-IR) spectra. The X-ray powder diffraction (XRD) analysis shows a pattern corresponding to stoichiometric hydroxyapatite (HA) with broad peaks suggesting that CDHA particles were nanosized. Fourier transform Raman spectroscopy (FT-Raman) do not indicate any fluorescence band that is characteristic of non-stoichiometric HA. The thermal decomposition of CDHA to beta tricalcium phosphate (beta-TCP) was also studied for the additional confirmation. The nanosized CDHA was found to be stable up to 600 degrees C. PMID- 15747181 TI - Synchronous delivery systems composed of Au nanoparticles and stimuli-sensitive diblock terpolymer. AB - A method to construct synchronous delivery systems via direct self-assembly of Au nanoparticles on the poly[(N-isopropylacrylamide-r-acrylamide)-b-L-lactic acid] (PNAL) nanospheres has been presented in this paper. To achieve amphiphilic diblock terpolymer, hydrophobic poly (L-lactic acid) (PLLA) block was added to poly(N-isopropylacrylamide- r-acrylamide) (PNA) block via Michel-type addition reaction. Lower critical solubility temperature (LCST) was modulated at 35.6 degrees C which is close to the body temperature, but higher than poly(N isopropylacrylamide) (PNIPAAm) homopolymer by controlling the ratio between isopropylacrylamide (IPAAm) monomers and acrylamide (AAm) monomers. Using this amphiphilic diblock terpolymer, PNAL nanospheres were fabricated by emulsion/evaporation technique followed by direct self-assembly of Au nanoparticles on the PNAL nanospheres due to the high affinity of amino groups donated from PNA block. The 'core' site of Au@PNAL nanospheres can load various lyphophilic drugs. Moreover, Au nanoparticles in the 'shell' domain of PNAL nanospheres give optimal environment to conjugate various biomolecules. Therefore, it is expected that Au@PNAL hybrid nanospheres can be utilized in synchronous delivery of both biomolecules in the 'shell' domain and various therapeutic drugs in the 'core' domain. PMID- 15747182 TI - In vitro study on bone formation and surface topography from the standpoint of biomechanics. AB - Effect of surface topography upon cell-adhesion, -orientation and differentiation was investigated by in vitro study on cellular responses to titanium substratum with different surface roughness. Cell-shape, -function and differentiation depending upon the surface topography were clarified by use of bone formative group cells (BFGCs) derived from bone marrow of beagle's femur. BFGCs consisted of hematopoietic stem cells (HSC) and osteogenetic stem cells (OSC). Cell differentiation of BFGCs was expressed and promoted by structural changes of cytoskeleton, and cell-organella, which was caused by mechanical stress with cytoplasmic stretching of cell adhesions to the substratum. Phagocytic monocytes of HSC differentiated to osteomediator cells (OMC) by cytoplasmic stretching with cell adhesion to the substratum. The OMC mediated and promoted cell differentiation from OSC to osteoblast through osteoblastic phenotype cell (OBC) by cell-aggregation of nodules with "pile up" phenomenon of OBC onto OMC. The osteogenesis might be performed by coupling work of both cells, OMC originated from monocyte of HSC and OBC originated from OSC, which were explained by SEM, TEM and fluorescent probe investigation on BFGCs on the test plate of cp titanium plates with different topographies. This osteogenetic process was proved by investigating cell proliferation, DNA contents, cell adhesion, alkaline phosphatase activity and osteocalcine productivity for cells on the titanium plates with different topographies. The study showed increased osteogenic effects for cells cultured on Ti with increased surface roughness. Possible mechanisms were discussed from a biomechanical perspective. PMID- 15747183 TI - Morphological evaluation of bioartificial hydrogels as potential tissue engineering scaffolds. AB - Poly(vinyl alcohol) hydrogels prepared by freeze-thawing procedure represent synthetic systems widely investigated as non-biodegradable scaffolds for tissue regeneration. In order to improve the biocompatibility properties of pure poly(vinyl alcohol) (PVA) hydrogels, blends of PVA with different biological macromolecules, such hyaluronic acid, dextran, and gelatin were prepared and used to produce "bioartificial hydrogels". The porosity characteristics of these hydrogels were investigated by scanning electron microscopy and mercury intrusion porosimetry. The morphology of bioartificial hydrogels was evaluated and compared with that of pure PVA hydrogels. In particular the effect exerted by each biological component on pore size and distribution was investigated. The obtained results indicate that when a natural macromolecule is added to PVA the internal structure of the material changes. A small amount of biopolymer induces the structural elements of PVA matrix to take on a well evident lamellar appearance and an apparent preferential orientation. Comparing the results of SEM and mercury intrusion porosimetry it was concluded that hydrogels containing 20% of biological component have the most regular structure and at the same time the lowest total porosity. On the contrary samples with the highest content of natural polymer (40%) show the less regular structure and the highest total porosity. PMID- 15747184 TI - Porous and dense poly(L-lactic acid) and poly(D,L-lactic acid-co-glycolic acid) scaffolds: in vitro degradation in culture medium and osteoblasts culture. AB - The use of bioresorbable polymers as a support for culturing cells has received special attention as an alternative for the treatment of lesions and the loss of tissue. The aim of this work was to evaluate the degradation in cell culture medium of dense and porous scaffolds of poly(L-lactic acid) (PLLA) and poly(D,L lactic acid-co-glycolic acid) (50:50) (PLGA50) prepared by casting. The adhesion and morphology of osteoblast cells on the surface of these polymers was evaluated. Thermal analyses were done by differential scanning calorimetry and thermogravimetric analysis and cell morphology was assessed by scanning electron microscopy. Autocatalysis was observed in PLGA50 samples because of the concentration of acid constituents in this material. Samples of PLLA showed no autocatalysis and hence no changes in their morphology, indicating that this polymer can be used as a structural support. Osteoblasts showed low adhesion to PLLA compared to PLGA50. The cell morphology on the surface of these materials was highly dispersed, which indicated a good interaction of the cells with the polymer substrate. PMID- 15747185 TI - Diamond coated dental bur machining of natural and synthetic dental materials. AB - Diamond coatings are attractive for cutting processes due to their high hardness, low friction coefficient, excellent wear resistance and chemical inertness. The application of diamond coatings on cemented tungsten carbide (WC-Co) burs has been the subject of much attention in recent years in order to improve cutting performance and tool life. WC-Co burs containing 6% Co and 94% WC with an average grain size 1-3 micron were used in this study. In order to improve the adhesion between diamond and the bur it is necessary to etch away the surface Co to prepare it for subsequent diamond growth. Hot filament chemical vapour deposition (H.F.C.V.D.) with a modified vertical filament arrangement has been employed for the deposition of diamond films. Diamond film quality and purity has been characterised using scanning electron microscopy (S.E.M.) and micro-Raman spectroscopy. The performance of diamond coated WC-Co burs, uncoated WC-Co burs, and diamond embedded (sintered) burs have been compared by drilling a series of holes into various materials such as human teeth, and model tooth materials such as borosilicate glass and acrylic. Flank wear has been used to assess the wear rates of the burs when machining natural and synthetic dental materials such as those described above. PMID- 15747186 TI - Preparation and characterization of a novel bioactive bone cement: glass based nanoscale hydroxyapatite bone cement. AB - A novel type of glass-based nanoscale hydorxypatite (HAP) bioactive bone cement (designed as GBNHAPC) was synthesized by adding nanoscale hydroxyapatite (HAP) crystalline (20-40 nm), into the self-setting glass-based bone cement (GBC). The inhibition rate of nanoscale HAP and micron HAP on osteosarcoma U2-OS cells was examined. The effects of nanoscale HAP on the crystal phase, microstructure and compressive strength of GBNHAPC were studied respectively. It was concluded that nanoscale HAP could inhibit the cell proliferation, while micron HAP could not, and that nanoscale HAP could be dispersed in the cement evenly and the morphology did not change significantly after a longer immersion time. XRD and FTIR results show nanoscale HAP did not affect the setting reaction of the cement. Furthermore, GBNHAPC had a higher compressive strength (92 MPa) than GBC. It was believed that GBNHAPC might be a desirable biomaterial that could not only fill bone defects but also inhibit cancer cell growth. PMID- 15747187 TI - The wear and fracture behaviour of ultra high molecular weight polyethylene subjected to gamma-irradiation in an atmosphere of acetylene. AB - The wear and mechanical properties of GUR 1020 (Perplas IMP 2000-2) Ultra High Molecular Weight Polyethylene (UHMWPE) subjected to gamma-irradiation in an atmosphere of acetylene, were evaluated for a range of processing conditions of irradiation, annealing and ageing. The results were compared with those obtained for the virgin UHMWPE material and material processed using conventional gamma irradiation in nitrogen. Cross-linking produced by irradiation in acetylene, followed by subsequent annealing was found to be significantly more effective in improving the mechanical and wear properties of UHMWPE compared to when the material was irradiated in nitrogen. Gel fraction analysis on its own, while being able to detect the degree of cross-linking, was found to be insufficient in determining the effectiveness of the cross-links and the resulting mechanical properties of the UHMWPE material. The results suggest that gamma-irradiation in an atmosphere of acetylene may provide significant advantages over conventional UHMWPE processing and irradiation cross-linking techniques. PMID- 15747188 TI - Release of chlorhexidine digluconate and flexural properties of glass fibre reinforced provisional fixed partial denture polymer. AB - The objective of this study was to determine the flexural properties and the release of chlorhexidine digluconate (CHX) of CHX laced unidirectional E-glass fibre reinforced provisional fixed partial denture polymer. Bar shaped test specimens (3.3 x 10.0 x 65.0 mm) were fabricated from provisional fixed partial denture polymer (mixture of poly[ethylmethacrylate] powder and n-poly[butyl methacrylate] monomer liquid) with E-glass fibre reinforcements. Poly(methyl methacrylate) preimpregnated continuous unidirectional glass fibre reinforcement was laced with CHX. The glass fibre reinforcements were incorporated into the polymer and the polymerised to the form of test specimens. In addition test specimens without CHX in glass fibre reinforcement were made for comparison. Control specimens did not contain glass fibres in the test specimens. Flexural strength and modulus of test specimens (n = 6) was tested with three-point bending test after storing the specimens dry or in water (two weeks). Released CHX was determined with high performance liquid chromatography during 180 days water immersion. In dry conditions, the flexural strength and the modulus of the polymer was 43 MPa and 1.7 GPa, and with glass fibre reinforcement 96 MPa and 3.5 GPa. With the reinforcement laced with CHX, the strength was 92 MPa and the modulus was 3.2 GPa. The water storage of test specimens did not weaken the reinforced polymer. The majority of the CHX released from the glass fibre reinforced polymer during the first days of storage in water. Flexural properties of provisional fixed partial denture polymer were increased using glass fibre reinforcement. The fibre reinforcement that was laced with CHX resulted in similar reinforcing effect. PMID- 15747189 TI - Ki67, HSP70 and TUNEL for the specification of testing of silicone breast implants in vivo. AB - This investigation of capsular tissue adjacent to silicone breast implants concerns the long-term tissue response to the implant environment. Fifty-three silicone breast implants have been analyzed at the time of explantation. The implant duration ranged from 2 months to 153 months. The reason for explantation was capsular contracture (57%), dissatisfaction with the effect (11%), local inflammation (6%), implant rupture (4%) and exchange of tissue expanders (21%). The cell turnover within the interface of the silicone device and the fibrous capsule was detected by specific antibodies against Ki67 for cell proliferation, by TUNEL for apoptosis, and by DNA strand breaks and heat shock protein 70 (HSP70) for cell stress. We found a negative correlation between the expression of HSP 70 and the capsular thickness (p < 0.043) and decreased levels in specimens obtained from Baker IV implant capsules. Ki67, and TUNEL were significantly positive (p < 0.001 for both) and HSP 70 were significantly negative (p < 0.001) with signs of inflammation. Both Ki67 and TUNEL indicated decreasing values over time. Ki67 and TUNEL showed no correlation with clinical signs of implant failure, such as the Baker score. The expression of HSP70, on the other hand, was connected with structural changes of the implant capsule, in terms of capsular thickness and the Baker score. PMID- 15747190 TI - [Health--economy and fairness]. PMID- 15747191 TI - [Promoting rehabilitation via research and evidence-based medicine]. PMID- 15747192 TI - [Practical social medicine and rehabilitation--on the development and challenges of a specialty department]. PMID- 15747193 TI - [Requirements of a future-oriented social medicine]. AB - With the new national licensing regulations for physicians subsections of the social medicine became discrete subjects. The question arises, which contents the social medicine can have in the future, with consideration of important basic conditions. Such are the progress of medical knowledge, the representation of social medicine at medical faculties, changes of the medical supply, the transformation of jobs and the globalization. On a long-term basis effects of the demographic development, changes of the family structure and the financing of health and illness are important too. The social medicine should promptly make quality-assured contents available with consideration of the Internet. Such contents could be the comprehensive consultation, investigation and control of patient careers as well as the consultation and investigation from health problems in municipalities and in the society. In addition an inductive and practical oriented curriculum should be compiled, using the subject catalogue of the social medicine as well as a new basic textbook of social medicine. PMID- 15747194 TI - [The changing role of the state in health care systems in OECD countries- research questions, research design and first results]. AB - In the framework of the DFG-Research-Unit 597 "Transformation of the State" is Project C3 investigating the change of the role of the state in health care systems in OECD-countries between 1970 and 2000. The primary research hypothesis is, that a tendency of convergence exists regarding the role of the state for the three main dimensions of health care systems, namely service provision, financing and regulation. This is carried out firstly based on a quantitative oriented analysis for 23 OECD-countries, utilising the data set "OECD Health Data 2002", and secondly with qualitative oriented country-studies for England, Germany and the U.S. Findings for the qualitative country-studies give some hints of convergence regarding the "private-public-mix" between the health system types "social insurance" (Germany), "national health system" (England), and "market oriented health system" (USA). First quantitative results concerning the changing role of the state in relation to the health care systems in OECD-countries provide evidence for a convergence process between these states in the financing dimension. For the dependent variable "total health care costs in % of gross domestic product" a trend of convergence is observed for the members states of the European Union (n = 12), while in the remaining Non-EU countries (n = 11) a trend of divergence is identified. For the same parameter we found a convergence for OECD-countries of the type "national health system" (n = 14). For OECD countries of the type "social insurance" (n = 8) this convergence process is observed only for the time period 1970 - 1980. Some results confirm the hypothesis of the "growth to limits" (Flora). But it appears to us that in future other significant factors like globalisation processes, the increasing potentials of the medical technology and demographic changes may play an important role for the overall health budget and specific needs of the population in the OECD states. PMID- 15747195 TI - [The structurizing effect of minimum quantitative requirements for inpatient care]. AB - PURPOSE: German social law ( section sign 137 SGB V) authorized the partners in health care to establish and develop a catalogue of procedures for which quality is associated with volume. There is lack of evidence regarding the volume limits. Other aspects need to be taken into account, first of all by consultation of performance data analysis. METHODS: Inpatient-data of hospitals in Baden Wurttemberg were studied to make out possible effects of volume standard codes on health care provision. For each procedure studied an ICPM- tracer was defined and a volume limit fixed. RESULTS: If a limit affects many hospitals but a small portion of health care to be changed, it means, that only very low volume providers would be excluded. An adequate network of care can be achieved by joining volumes from neighbouring regions. The analysis showed that 39 % of the regions offering pancreatectomy come to a minimum volume of ten operations per year. A volume below 5 procedures per year is performed in 12 regions. If these small amounts are brought together, about two thirds of the regions involved in this segment of care could continue with a local offer corresponding to that defined standard. For certain high-risk conditions or procedures centralisation has already taken place (e. g. coronary bypass surgery). CONCLUSIONS: Based on appropriate analysis of the data of all partners in health care concepts can be tailored to the needs of the statutory health care system and accompanying activities involved from the very beginning. Risks have to be taken into consideration, including rarefaction of health care offers, disparities in care and on the other hand economically motivated decentralization, where risk adapted centralisation was necessary. PMID- 15747196 TI - [What do we conclude from re-admissions about quality of inpatient care?]. AB - Quality assurance in hospital care increasingly focuses on evaluation of outcome. Problems arise with displaying results of medical care beyond discharge. In this context hospital readmissions are often used as outcome variable. But it is unclear whether readmissions are meaningful indices of quality of hospital care and if so, where to get valid data on readmissions. We used claims data of the regional health insurance fund in Saxony-Anhalt (AOK Saxony-Anhalt) from 2002 and 2003 (850,000 insured; nearly 300,000 cases per year). All hospital admissions of a insured person are identified by an anonymous id-number independent of the admitting hospital. By this way we can analyze readmissions individually. Readmission are frequent events in hospital care. Nearly one third of all patients were admitted at least a second time in 2003. 18 % of all hospital cases are readmissions within 30 days after discharge. Readmissions concentrated on chronically ill, oncological, or multimorbid patients. Many of the readmissions take place in the context of planned therapies or post-operative treatment. 'Revolving-door patients' with multiple readmissions point to problems in cooperation of ambulatory and hospital care. By defining tracer diagnoses and specific causes of readmissions unplanned readmissions may be identified as a quality indicator of suboptimal care. Readmissions don't express suboptimal care per se. But taking into account methodological aspects a tracer approach with defining specific unplanned readmissions may provide meaningful outcome indicators. These can be derived from claims data fast, routinely, and with low costs. Further validation of the approach is needed. PMID- 15747198 TI - [Health promotion aspects in reorganization of a centralized operating department]. AB - In many health care institutions (eg, hospitals), employees' health and satisfaction are not the main concern. However, if health and productivity are affected by high sickness rates and low motivation, health and satisfaction of the employees become important. This applies especially to hospitals engaged in acute patient care. We showed that a complete reorganisation of the central O. R. resulted in employees' working satisfaction, low illness rates (- 70 %) and increased motivation. In addition, the new structure increased the efficacy (> 500 operations more per year), improved the economy (savings of 500,000 annually) and increased patient satisfaction as expected and desired. PMID- 15747197 TI - [Efficiency of general practice in Switzerland]. AB - BACKGROUND: In their capacity as gatekeepers, affect the entire cost of treatment of patients including cost for medication and inpatient care. Thus, it is of special interest to examine the production process of general practitioners. AIM: To study efficiency differences between general medical practices, taking into account a possible Managed Care participation by the physician. METHODS: A stochastic frontier analysis based on billing data of 1,500 Swiss physicians from 1997 - 1999 is applied to measure the relative efficiency of general practitioners. The number of consultations serves as output and the charged services as inputs of production. RESULTS: Most productive physicians are members of primary physicians' networks, in which the gate-keeping function of general practitioners is of central importance. For the "production" of one consultation they use 15 percent less resources than an average physician in a conventional medical practice. DISCUSSION AND CONCLUSION: In principle, assessment of efficiency of primary care physicians is possible. Differences in efficiency between physicians can be explained by characteristics of the individual medical practice. Disadvantages of the approach exist in the neglect of the patients' severity of illness and in having indirect measures of quality of medical care available. PMID- 15747199 TI - [Strategic controlling in the consideration of social demographic changes in social medical services]. AB - Annually the Medizinische Dienst der Krankenversicherung Westfalen-Lippe (MDK-WL) performs approximately 120,000 nursing care assessments according to the 11th social statute book (SGB XI). A prospective reflection on the amount of expert assessments and the spectrum of services for the Nursing Care Insurance until 2020, was performed in order to establish a long term strategic controlling. The insured party makes the request for nursing care according to their personal estimation when the need for assistance is increasing. To predict the future amount of expert assessments you have to take into consideration the social background of the families in addition to age and gender (there is a clear correlation between age and the need of assistance). The database from nursing care assessments in 2001 was projected on a demographic model for the region of Westfalen-Lippe. The amount of requests correlates as expected with the patients age and increases exponentially. The incidence in the need of care shows relevant gender differences, but taken into consideration the very different age structure, the overall incidence is very similar. Against the background of the current nursing care law, the MDK-WL has to deal with an annual increase in assessments of 2 %. The requests for single persons are extremely often without foundation (55 % not substantially in need of care vs. 35 %). Looking at the requests of couples, it shows that the ones for women are more often unfounded then the ones for men (39 % vs. 32 %). It is necessary to take the development of the amount of single living persons into consideration to achieve more accurate predictions for the amount of assessments. PMID- 15747200 TI - [Perceptions of quality of life in schizophrenic patients]. AB - BACKGROUND: Subjective quality of life in schizophrenic persons is a commonly applied indicator in outcomes research. However, it remains unclear whether the content of the construct is adequately represented from the patients' point of view, and whether changing representations in the course of illness should be accounted for in terms of re-prioritization of the concept. PURPOSE: Identification of specific perceptions of quality of life (QoL) of schizophrenic persons and analysis of changes of those perceptions with increasing chronicity. METHODS: The present study is a reanalysis of responses to a short, structured, open-ended interview taken from different studies in social psychiatry research. 403 schizophrenic persons from different care settings (psychiatric hospitals, community based institutions) as well as 178 mentally healthy persons were interviewed with regard to positive and negative aspects of their QoL. A qualitative content-analysis resulted in a comprehensive hierarchy of categories. The proportion of persons who mentioned a respective QoL-aspect was compared. Relationship between chronicity and perceptions of QoL were analysed by means of chi (2)(trend)-tests. RESULTS: Schizophrenic persons mentioned about one fewer QoL domain compared to the mentally healthy (8.3 (sd = 2.8) vs. 9.6 (sd = 2.0), P < 0.001). They had fewer assertions on work aspects as well as in social domains (family, social relationships, partnership). The proportion of statements in social domains was negatively associated with the degree of chronicity. Among other things the schizophrenic persons showed a higher proportion of statements regarding "oral needs" (food, drinking-related statements, smoking) and (problems with) daily activities; both domains were associated with the degree of chronicity. DISCUSSION: Differences between schizophrenic and mentally healthy persons in their perceptions of QoL were more pronounced with increasing chronicity. It remains unclear at which point in the course of illness this is best understood in terms of re-prioritization or even re-conceptualization of the entire concept. PMID- 15747201 TI - [Deficits and outlook on gender-specific health care as exemplified by KHK and mental disorders]. AB - Gender differences in health care are controversially discussed. Results show superfluous, insufficient or inappropriate health care of female and male patients. Many publications show that women are not sufficiently included in clinical trials and therefore data are not useful for analysing gender differences. Survey data like the German National Health Interview and Examination Survey 1998 (Bundes-Gesundheits survey 1998) can be used alternatively to analyse gender specific issues. This survey was used to show possible age and gender differences in prevalence, drug usage and health counselling of female and male patients with coronary heart disease and mental disorders. The number of physician attendances differs in sex and age group in both groups of illnesses. Women have higher number of physician consultations than men and the elderly more than the younger persons. For elderly women with coronary heart disease the control of blood cholesterol levels seems to be poorer than in men. Men get more health counselling from their doctors than women. The results emphasize the importance of considering gender differences in medical research. PMID- 15747202 TI - [Subjective health as predictor for mortality]. AB - In the last two decades self-rated health has received growing interest in international studies because of its consistent prediction for mortality. However, for Germany there are no studies confirming a long-term effect independent from objective health indicators in comparison of different follow up. On the basis of the Life-Expectancy-Study (1984/86 - 1998) from the Federal Institute for Population Research it was possible to analyze the association between subjective health and mortality in relation to the length of observation. A stronger correlation between bad self-rated health and objective health status could be indicated because of a better prediction for mortality in a short-term follow-up. The evidence of a significant effect between self-rated health and mortality in the long-term follow-up not including the deaths from the short-term follow-up indicates that the mechanisms between subjective health and mortality are more complex than those between objective health status and death. PMID- 15747203 TI - [Learning healthy living--health management in schools. Health promotion in schools with particular consideration of socially disadvantaged students]. AB - "Learning to Live Healthy" is a cooperation project between the Central Association of Health Insurers and the Health Associations of Lower Saxony, Saxony-Anhalt and Rhineland-Palatinate. Our goal is to develop a healthy environment for all teachers and students in the "School Organization". Our focus lies -- more strongly than with earlier projects for school health promotion -- in the improvement of the health of teachers and non-teaching personnel. Not until their resources are strengthened and overwork/overload broken down, can resources be directed to health promotion for and with students, and health risks connected with social inequalities can be diminished. The project operates with the setting approach, and uses the principles of Workplace Health Management- Integration, Participation and Project Management. The project utilizes the instruments of project organization to jump-start the process of change in the School Organization, systematically, through goal-oriented action and sustainable development, thus improving the quality of education. PMID- 15747204 TI - [Subtainable health promotion via organisational development--a model project for teachers in professional training schools]. AB - The goal of this project is the development, implementation and evaluation of a concept designed for sustainable health promotion among occupational and trade school teachers. We assume that for sustainable health promotion -- along with a behavioral prevention program -- a change is necessary in the structure, as well as, the working and communication processes within schools. The realization of early teacher participation and self regulated cooperative groups initiates comprehensive and goal-oriented developmental processes in the project schools. The organizational development process was accomplished in the following way: At the beginning we conducted a diagnosis of school-specific and individual health risks and the resources available to the project schools. The results were reported for both the individual and for the teacher group. This was intended to clarify the potential for improvement and, thus, strengthen the teachers' motivation toward processes of change. Following the diagnosis, the teachers chose areas of stress-related strain and then worked in groups to develop and implement behaviour and working condition-oriented intervention strategies for health promotion. The diagnosis results confirm the necessity of school-specific health promotion: the schools demonstrate very different demand and resource profiles. Furthermore, is has become evident that the central success factor for health promotion in schools is the teachers' willingness for change. The individual and group reports of the diagnosis results seem to have made clear how essential individual and organisational changes are. PMID- 15747205 TI - [Area-wide dental prophylaxis methods and locating increased needs for prophylaxis in Braunschweig 2003/2004]. AB - Based on the section sign 21 SGB V an extensive concept for measuring tooth prophylaxis methods is used in Braunschweig. The program attains 20,000 children at the age of 3 to 15 years. It is mainly financed by the working pool of the health insurance. Small spatial gathered indicators are visualized in 30 defined parts of the town. City maps and excel-tables are presented in a SWOT-analysis by the example of the data of the primary schools. PMID- 15747206 TI - [Integrated sustainability-oriented reporting--key indicators for communities and cities. Results of a research and development project]. AB - In our research project -- supported by the German Federal Ministry for Education and Research (BMBF) - 35 key indicators were developed in an ongoing process in co-operation with 10 East German cities, which are all members in the German cities healthy network. With these indicators the cities should take up integrated sustainability - oriented health reporting, from which actions and projects for health promotion and prevention can be derived. The conceptual background for the research project and the reports to be made by the project in co-operation with the cities, are the three policy programmes that to be realised on a county level: Healthy Cities, Local Agenda 21 and the German city development programme "Soziale Stadt (socially oriented city)". The common goal of these programmes is the sustainable improvement of the quality of life in the counties. The project is part of the BMBF-supported research field "problem - oriented regional reporting systems", in which other projects are involved which are mainly being conducted in the newly-formed East German "Lander". In this article we describe the co-operative process of the development of the indicators. A synopsis of already applied or proposed sets of indicators for municipal reporting was the basis for the development of the project's set of 35 key indicators. The set of indicators is presented according to its usefulness for planning and realisation of actions for health promotion on county level. For each of the 35 indicators a meta - data description was made to support the counties and cities in our project for health reporting. These indicator profiles are also helpful and supportive for all counties and citiesaiming at such health reporting. The project started in May 2002 and lasts till Mai 2005, so that most reports should be completed in the spring of 2005. PMID- 15747207 TI - [Missing data in the somatisation subscale of the scl-90-R due to a 'checklist effect': occurrence in different surveys and results of various imputation methods]. AB - Item non-response is a potential threat to the validity of study results. Taking the somatisation subscale of the SCL-90-R as an example, we hypothesise a specific response pattern ("checklist-effect") that is characterised by symptom free persons not checking the "not at all"-category. The present study analyses the extent and relevance of this postulated "checklist-effect". Our data is derived from a survey of n = 228 blue-collar workers who previously had filed applications for medical rehabilitation benefits (A1-study), and two additional surveys as well. We defined the "checklist-effect" by the following response pattern: (1) at least one missing value and (2) at least one valid item response and (3) no "not at all"-responses. Occurrence of the "checklist-effect" in the three datasets differed widely. 75 % of the responders in the A1-Study had complete data, 16.2 % a postulated "checklist-effect". Imputation of missing values under the assumption of a "checklist-effect" led to a reduction of missing data in the somatisation-subscale from 12.3 % to 0.4 %. Ignoring the "checklist effect" would overestimate the symptom level. Possible explanations for the effect are discussed. However, the validity of this effect has yet to be proven via methodological studies. PMID- 15747208 TI - [Assessment of cost-benefit analysis in integration projects on drug addiction]. AB - The purpose of this study is to make a cost-benefit-analysis for integration projects helping drug addicts and substitutes to reintegrate into society. The study is intended to contribute to a better allocation of resources under the trade-off-situation that only a limited number of integration projects can be realized due to budget limitations. This pilot study represents an economic evaluation of health activities on integration based on the example of study projects offered by Mudra e. V. As a result the study showed that the evaluated projects are economically advantageous during the investigated research period. Furthermore, the study contains a non-monetary analysis of intangible effects which shows significant improvements in quality of life. Although the results are substantial, further research is mandatory focussing on the economic benefits of integration projects. PMID- 15747209 TI - Deep brain stimulation in intraoperative MRI environment - comparison of imaging techniques and electrode fixation methods. AB - We performed 118 consecutive DBS cases from November 1999 to June 2002. Intraoperatively there were 10 cases studied with fluoroscopy, 73 with 0.2 Tesla (T) MRI and 35 with 1.5 T MRI. Ten electrodes were secured by Medtronic caps, 25 by methyl methacrylate with titanium miniplates, and 82 by Navigus caps. The 3 dimensional displacement between the planned target and actual electrode position (3DD) was determined by fusing the postoperative MRI with the preoperative imaging. The 3DD for using Medtronic caps, methyl methacrylate with miniplates, and Navigus caps were 4.80 +/- 3.16, 2.64 +/- 1.26 and 2.23 +/- 1.15 mm (mean +/- SD), respectively. Navigus caps had statistically significant accuracy (P = 0.03) in holding the electrode when compared with Medtronic caps, and it facilitated electrode revision. The fixation devices significantly affect the final vertical position of the electrode. The 3DD for fluoroscopy, 0.2 T and 1.5 T MRI cases were 4.80 +/- 3.16, 2.31 +/- 1.21 and 2.34 +/- 1.14 mm (mean +/- SD), respectively. No statistically significant difference (P = 0.91) in 3DD was demonstrated between 0.2 T and 1.5 T MRI cases. The presence of intraoperative 1.5 T MRI allowed near real-time electrode position confirmation and early detection of hemorrhagic complications. Satisfactory microelectrode recording was feasible in low-field 0.2 T and high-field 1.5 T MRI environments. Further studies on performing DBS in real-time intraoperative MRI are warranted. PMID- 15747210 TI - Neuronavigation in skull base tumors. AB - OBJECTIVE: Computer-assisted neuronavigation was used in 87 cases of skull base lesions (SBLs). Preoperative planning and intraoperative identification of anatomic landmarks is especially important in SBLs since it helps to avoid or minimize surgical morbidity and mortality. In this study, we assessed the accuracy and the clinical usefulness of a frameless system based on the optical digitizer in SBLs. PATIENTS AND METHODS: Between April 2000 and March 2003, eighty-seven patients with SBLs were operated on in our department using cranial neuronavigation. A passive-marker-based neuronavigation system was used for intraoperative image guidance. There were 56 women and 31 men. The patient's ages ranged from 4 to 76 years (average: 45.7 year). The locations of the tumors reported in this series were as follows: frontobasal, 24 cases; sellar/parasellar, 32 cases; petroclival, 16 cases; tentorial/subtemporal, 15 cases. RESULTS: The computer-calculated registration accuracy ranged between 0.3 and 1.7 mm (mean, 1.1 mm). Gross total removal of the SBLs was accomplished in 82 out of 87 patients as was confirmed on postoperative CT and MRI scans. The follow up period ranged from 1 month to 48 months (average: 20.1 months). Overall mortality and severe morbidity (meningitis, permanent cranial nerve deficits, and cerebrospinal fluid fistulae) rates were 4.6 % and 33.3 %, respectively. CONCLUSION: The image-guided surgery is a valuable aid for safe, helpful and complete removal of SBLs of the brain where accurate localization of the lesion is critical. Although our preliminary series is not large, interactive image guidance provides a constant display of surgical instrument position during surgery and its relationship with the SBLs components, surrounding normal brain, and vascular structures, providing valuable guidance to the surgeon during an operation. Our experience with the neuronavigation suggests that image guidance is helpful in this type of lesions, providing better anatomic orientation during skull base surgery, delineating tumor margins and their relation to critical neurovascular structures. PMID- 15747211 TI - Advanced neuronavigation in skull base tumors and vascular lesions. AB - OBJECTIVE: The purpose of this study was to describe the usefulness of recent advances of neuronavigational technology in the management of skull base tumors and of vascular lesions, treated via a skull base approach. METHODS: In 16 patients (skull base meningioma n = 9, petrous apex epidermoid n = l, craniopharyngeoma n = 1, giant internal carotid artery aneurysm n = 1, basilar/vertebral artery aneurysm n = 2, brain stem cavernoma n = 2), "advanced" neuronavigation was used. In contrast to "conventional" neuronavigation, the information for the neurosurgeon was enhanced by the intraoperative screen display of 3-dimensional reconstructions of the lesion, vessels, nerves and fiber tracts at risk. The 3-dimensional reconstructions were obtained by preoperative manual or automated segmentation processes. In addition, different imaging modalities (computed tomography [CT] with magnetic resonance imaging [MRI], CT with CT angiography, T (l)- with diffusion-weighted MRI) were fused and shown on the screen. RESULTS: In the cases of tumors, "advanced" neuronavigation facilitated the approach (n = 4), contributed to tailor the approach (n = 2) and helped to identify hidden neurovascular structures (n = 9). In the cases of aneurysms, "advanced" neuronavigation allowed us to reduce the skull base approach to the needs of safe aneurysm clipping (n = 3). In both cases of brain stem cavernoma, "advanced" neuronavigation was deemed useful for definition of the best surgical approach in relation to the pyramidal tract and brain stem nuclei. CONCLUSION: The authors' experiences suggest that neuronavigation, which displays 3-dimensional reconstructions of lesion, vessels, nerves and fiber tracts during surgery and makes use of image fusion techniques, is an important tool in the neurosurgical management of skull base lesions. PMID- 15747212 TI - Endoscopic neuroanatomy through the sphenoid sinus. AB - OBJECT: The aim of the study was to perform endoscopic anatomical studies of skull base structures through the sphenoid sinus in order to better understand endoscopic approaches to skull base lesions. METHODS: Anatomical studies were performed on six cadavers using sinus endoscopes. The sphenoid sinus was entered via a sublabial nasoseptal approach. Stepwise removal of bone surrounding the sphenoid sinus was done to expose the suprasellar area, the retrosphenoid area and the cavernous sinus. The suprasellar dissection exposed the anterior communicating artery (ACom) complex, optic apparatus, pituitary stalk, lamina terminalis, third ventricle and pituitary gland. The retrosphenoid dissection exposed the mamillary bodies, basilar artery and its branches, the mid-brain, the pons, and the third, fifth, sixth, seventh and eighth cranial nerves. The cavernous sinus dissection exposed the carotid artery, and the third, fourth, fifth and the sixth cranial nerves. CONCLUSION: Through the sphenoid sinus, most of the structures in the suprasellar area, cavernous sinus and retrosphenoid area can be reached. PMID- 15747213 TI - Surgical anatomy of the cribriform plate and adjacent areas. AB - Continuous improvement of surgical techniques is needed in skull base surgery in order to decrease intraoperative complications during various operations. However, the basis of minimizing damage in the operative field is a comprehensive knowledge of microsurgical anatomy. This article provides a detailed description of the cribriform plate and adjacent areas, based on investigations of over one thousand dried skulls and 225 computerized tomography scans of the anterior skull base. The main anatomic structures and the different types of the cribriform plate are demonstrated on color photographs and identified on CT scans as well. The variations important in anterior skull base approaches are also discussed and compared with the data found in the literature. The surface anatomy and development of the cribriform plate is described. PMID- 15747214 TI - Trans-eyebrow mini-orbitozygomatic pterional approach for minimally invasive skull base surgery. AB - Surgical treatment of skull base pathologies remains one of the most challenging tasks to neurosurgeons. Advances in neuroimaging as well as the evolution of modern techniques and neurosurgical equipment have paved the way for a more precise diagnosis, a better selection of the surgical approach and have prompted the adaptation of minimally invasive concepts to different aspects of modern neurosurgical practice. The trans-eyebrow mini orbitozygomatic pterional approach is a tailored approach that provides multiple avenues to the target, minimizing retraction of the brain and affords better exposure of the pathology. It was performed on 16 patients with frontobasal or suprasellar tumors. The post operative clinical outcome was excellent with smooth and fast recovery as well as satisfactory cosmetic results. It should be considered as part of the armamentarium to manage skull base lesions when indicated and not to replace other well recognized skull base approaches. PMID- 15747215 TI - Endoscope-assisted microsurgery of large vestibular schwannomas. AB - AIMS: The application of endoscope-assisted microsurgery in the treatment of small or medium-sized vestibular schwannomas is of proven value. The goal of our study is to evaluate its usefulness in cases of large schwannomas. PATIENTS AND METHODS: Eighteen patients were included in this prospective study. Their average tumor diameter was 3.9 cm. The retrosigmoid approach was used in all cases. The endoscope was applied during all stages of tumor removal. RESULTS: The facial nerve was visualized endoscopically at early stages of surgery in 9 patients and the abducent nerve in 7 patients. The source of bleeding was identified in 1 case. Tumor remnants in the region of the fundus of the internal auditory canal after apparently total removal were identified in 2 cases. Exposed and unobliterated temporal bone air cells were not observed. DISCUSSION: Even in cases of large schwannomas, the location of the facial nerve can be determined endoscopically early in the procedure. The application of endoscope-assisted microsurgery increases the rate of cranial nerve preservation and of total tumor removal. Although the application of the endoscope did not provide useful information in some cases, it is a safe procedure that did not lead to any complications and/or to considerable prolongation of the operative time. Its application is justified in all cases. PMID- 15747216 TI - Single burr hole surgery for the spheno-orbital fibrous dysplasia using intraoperative computed tomography. AB - Total removal of spheno-orbital fibrous dysplasia was achieved through intraoperative CT-assisted surgery via a burr hole. A 32-year-old man had persistent headache. Radiological studies demonstrated a small osteolytic lesion in the sphenoidal bone underneath the superior orbital fissure. Intraoperative serial CT scans showed the depth and width of the tumor within the complicated structure of the skull base. The lesion was successfully removed by CT-guided minimally invasive surgery. PMID- 15747217 TI - Endoscopic third ventriculostomy in post-tubercular meningitic hydrocephalus: a preliminary report. AB - Hydrocephalus is a common sequel of tubercular meningitis. Endoscopic third ventriculostomy (ETV) was performed in thirty-five patients. According to the duration of illness, six patients were in the early (less than 6 weeks), nineteen were in the intermediate (6 weeks to 6 months) and ten patients were in the late phase (more than 6 months) of tubercular meningitis (TBM). Six patients were in stage I, seven patients in stage II and twenty-two patients were in stage III. The overall success rate of ETV in TBM was 77 %. Sixty percent had early and seventeen percent had delayed recovery. Obstructive hydrocephalus was present in 54.3 % and 45.7 % had communicating hydrocephalus. The radiological recovery rate was 55.6 %. The outcome with a thin to transparent floor of the third ventricle was 87 %. PMID- 15747218 TI - Rescue use of endovascular snare for acute basilar artery embolic occlusion resistant to balloon angioplasty and fibrinolysis therapy. AB - A rescue clot disruption using a basket snare is described for acute basilar artery embolic occlusion resistant to balloon angioplasty and fibrinolysis therapy. In spite of failed balloon angioplasty in conjunction with fibrinolysis, a basket-shaped snare connected to a microguide wire could be used to catch and crush the clot in the upper basilar artery. The rescue use of a snare may be effective for angioplasty-resistant acute embolic stroke. PMID- 15747219 TI - Successful surgical excision of juvenile-type spinal arteriovenous malformation in two stages following partial embolization. AB - The case of a 24-year-old man with a juvenile-type spinal arteriovenous malformation is reported. Spinal angiography and magnetic resonance image revealed a juvenile (Type III) AVM at the cervical spine. This spinal malformation was successfully resected with a posterior and anterior surgical approach in two stages following partial embolization. PMID- 15747220 TI - Use of a nitrogen arm-stabilized endoscopic microdriver in neuroendoscopic surgery. AB - Stabilization and manipulation of instruments in endoscopic neurosurgery has traditionally been a manual task. Conventional self-retaining retractors have failed to provide reliable fixation for endoscopic procedures. Most surgeons rely on a team approach to endoscopy with one individual controlling the endoscope while the other manipulates instruments within the field of view. This two surgeon approach is manpower intensive and not an option at many facilities. Recently developed technology provides the independent practitioner with another option. Several nitrogen-powered stabilizing arms have been introduced which decrease or eliminate problems with post-positioning drift. To facilitate fine maneuvering of the endoscope, a new positioning arm is available with an incorporated microdriver capable of sub-millimeter adjustments. We present an illustrative case of the utility of this device in selected endoscopic procedures. PMID- 15747221 TI - [Neurophilosophy, neuroscience and psychopathology]. PMID- 15747222 TI - [The current mind-brain theories in analytical philosophy of mind and their epistemic significance for psychiatry]. AB - This article begins with an orientational survey of the historical evolution of analytical philosophy of mind (APM) which was formulated in the last 40 years as "philosophy of mind" in the Anglo-Saxon scientific-cultural world and which, in the meantime, dominates to a great extent contemporary German philosophy. Then there follows a discussion of the currently most popular mind-brain theories in philosophy. In comparison to the more marginal dualist variants (interactionism, epiphenomenalism, parallelism), it is mainly the monistic positions of non reductive, reductive and eliminative materialism and the materialist functionalism underlying it, which determines analytical philosophy of mind and its influence on psychopathology and psychiatry. Under the additional influence of modern brain research methods, particularly neuroimaging, it is progressively developing into a subdiscipline of neuroscience, a complex and increasingly more firmly established scientific discipline which comprises the totality of all sciences dealing with neuronal functions, including the close epistemic associations of APM and neuroimaging. This is the effective epistemic central idea determining the theory of the neuronal network which, in the form of a connectionist psychopathology, is intended to make possible a fundamentally new access to the comprehension of psychiatric forms of illness. In this respect it is evident, however, that the perception of the naturality of the mind as the fundamental thesis of APM and thus of connectionism cannot be followed through, since, up to now, neither from the phenomenality of the mind (especially the quality of senses, "Qualia") nor from intentionality of the mind (i. e. the ability to act intentionally, free from the constraint of the causality of nature and thus in self-responsible fashion) has proved it possible to reconstruct a generally accepted naturalist theory. Furthermore, it has not been possible to reformulate it in an exclusively physical, i. e. non-phenomenological concept and terminology which is, above all, free from the intentionality idiom. The consequence of this, however, is that a connectionist psychopathology can only represent a subpersonal, i. e., subhuman area and that in order to establish a personal psychopathology, naturalistic unreduced theories of experience-qualities and intentional acts of completeness are absolutely essential. The neuroscientific-connectionist paradigm of psychopathology must therefore - at least for the present - be supplement by the paradigm of a non-natural (e. g. phenomenological-hermeneutic) psychopathology. This result can only encourage the relinquishing of epistemically one-sided materialist and other monistic mind brain theories of APM in favour of an epistemically open pragmatic interactionist dualism as the scientific position which best represents the current state of knowledge. PMID- 15747223 TI - [Differential influence of immune therapy on relapses and progression in multiple sclerosis: interpretation and therapeutic consequences]. AB - It is well established that relapses can be suppressed by different substances in patients with relapsing-remitting multiple sclerosis (MS). In contrast, patients with progressive forms of MS do hardly respond to immune therapy. Therefore, start of immune therapy after the first relapse has been proposed, especially in order to prevent degeneration and disability. This view is challenged in the present review. Actually no evidence exists in support of a retardation or an attenuation of secondary progression by early immune therapy. Widespread degeneration occurs early and progresses independently from inflammatory plaques. Therefore, autoimmunity per se is no adequate paradigm to explain MS-pathogenesis entirely. A virus/superantigen-dualism is proposed to explain the different parts of MS, instead. It is concluded that evidence-based immune therapy should be adapted to the actual inflammatory activity of the disease. A suitable parameter for this purpose is the interval between 2 relapses. PMID- 15747224 TI - [Catamnestic study on the efficacy of an intensive outpatient treatment programme for alcohol-dependent patients: impact of participation in AA on the abstinence rates]. AB - This study analyzed the Alcoholics Anonymous participation and the impact on the abstinence rate of 103 alcohol dependent patients (ICD-10) during the 24 months after their discharge from high-structured out-patient treatment. The treatment retention amounted to n = 74 (72 %), 18 of the 25 dropouts took place because of alcohol relapse. At 6-, 12- and 24-months-follow-up 87 - 95 % of the patients were successfully located and re-interviewed. Analyses revealed that 64 % of the patients were abstinent at the 6-months-follow-up evaluation, 56 % at the total 12-months-follow-up evaluation. 49 % of the patients remained abstinent until the 24-months-follow-up evaluation, 14 % were improved and 37 % relapsed. 56 patients (55 %) participated in selfhelp-groups the first six months following treatment, two years later 45 patients (44 %) still attended a group. 53 - 56 % participated on a weekly basis. Patients who participated regularly on a weekly basis in self help-groups fared the best on 2-year outcome. Patients who infrequently at all participated or not had the poorest outcome (relapse). After controlling for confounding variables (sex, treatment drop-out, relapse during treatment) these results were still statistical significant. Results indicate the predictive value of AA attendance for relapse prevention after controlling for confounding variables. The value of self-help-groups in the network of alcoholism treatment is discussed. PMID- 15747225 TI - ["If Berger had survived the second world war - he certainly would have been a candidate for the Nobel Prize". Hans Berger and the legend of the Nobel Prize]. AB - The public opinion pays much attention to the Nobel Prize as an indicator for the scientific efficiency of a university or a country in connection with foundation of so-called elite universities. The former holder of the psychiatric chair in Jena and discoverer of the electroencephalogram Hans Berger (1873 - 1941) came into discussion as candidate for the Nobel Prize in physiology or medicine. The current medical-historical publications maintain the view that Berger should have received the Nobel Prize in 1936 as well as in 1949. This was prevented in 1936 by an enactment from Hitler, which forbid him to accept the prize, and later in 1949 by Berger's own death. According to documents of the Nobel archives these statements can be disproved. Berger was only nominated three times out of 65 nominations in 1940. Because of his death the other two recommendations in 1942 and 1947 were never evaluated. PMID- 15747226 TI - [Longterm therapy of schizophrenia]. PMID- 15747227 TI - Distribution of norspermidine as a cellular polyamine within micro green algae including non-photosynthetic achlorophyllous Polytoma, Polytomella, Prototheca and Helicosporidium. PMID- 15747228 TI - Comamonas badia sp. nov., a floc-forming bacterium isolated from activated sludge. AB - Strain IAM 14839, isolated from activated sludge in Japan, forms a visible floc and grows in the flocculated state. This bacterium is Gram-negative, rod-shaped, strictly aerobic and highly motile with a single polar flagellum. Both oxidase and catalase activities are positive. No growth was observed on sugars. The strain can grow at 20 degrees C, but does not grow at 37 degrees C. The G+C content of DNA is 66.3 mol% and Q-8 is the major quinone. The major cellular fatty acids are 16:1omega7c, 16:0, 18:1omega7c, 2OH 16:0, 3OH 10:0. The 16S rDNA sequence analysis indicated that the bacterium clustered within the genus Comamonas. On the basis of the phylogenetic analysis and phenotypic properties, it is proposed that the strain IAM 14839T be classified in a novel species of the genus Comamonas, Comamonas badia sp. nov. The type strain is IAM 14839T (=KCTC 12244T ). PMID- 15747229 TI - Kaistia adipata gen. nov., sp. nov., a novel alpha-proteobacterium. AB - A taxonomic study was carried out on Chj404T, a bacterial strain isolated from a soil sample collected in an industrial stream near the Chung-Ju industrial complex in Korea. The strain was a gram-negative, aerobic, short rod to coccus shaped bacterium. It grew well on nutrient agar medium and utilized a broad spectrum of carbon sources. The G+C content of the DNA was 67.4 mol% and the major composition of ubiquinone was Q-10. The major fatty acid was C18:1. Comparative 16S rDNA studies showed a clear affiliation of this bacterium to alpha-Proteobacteria. Comparison of phylogenetic data indicated that it was most closely related to Prosthecomicrobium pneumaticum (92.7% similarity in 16S rDNA sequence). Since strain Chj404 is clearly distinct from closely related species, we propose the name Kaistia adipata gen. nov., sp. nov. for this strain Chj404T (=IAM 15023T=KCTC 12095T). PMID- 15747230 TI - Distribution of two triamines, spermidine and homospermidine, and an aromatic amine, 2-phenylethylamine, within the phylum Bacteroidetes. AB - Cellular polyamines of the newly additional 19 species belonging to the class Bacteroides of the phylum Bacteroidetes were analyzed by HPLC to display polyamine distribution as a chemotaxonomic marker within the total 41 species. Three profiles, the presence of spermidine, the presence of homospermidine and the absence of both triamines, corresponded to their phylogenetical positions within the four families of the class. The occurrence of an aromatic amine, 2 phenylethylamine, extracted into cellular polyamine fraction, was also determined within the 121 species distributed within the phylum. This aromatic amine was found in Cellulophaga lytica, Cytophaga latercula, Tenacibaculum amylolyticum, Tenacibaculum martimum, Tenacibaculum mesophilum and Psychroflexus torquis belonging to the family Flavobacteriaceae of the class Flavobacteria, and Flexibacter flexilis and Microscilla marina belonging to the family Flexibacteraceae of the class Sphingobacteria. PMID- 15747231 TI - Candida easanensis sp. nov., Candida pattaniensis sp. nov. and Candida nakhonratchasimensis sp. nov., three new species of yeasts isolated from insect frass in Thailand. AB - Six strains of anamorphic yeasts isolated from insect frass collected in several regions of Thailand were assigned to the genus Candida based on the conventional taxonomic criteria used for yeast classification. These strains have Q-7 as the major ubiquinone and are suggested to have close relationships to the genus Pichia. Three strains, ST-225, ST-228 and ST-229, have identical nucleotide sequences in the D1/D2 domain of 26S rDNA and are closely related to Pichia japonica, but differ by six nucleotides (1.1% ) from this species. These three strains are considered to represent a single new species, which is described as Candida easanensis sp. nov. Two strains, ST-311 and ST-320, have identical sequences in the D1/D2 domain and resemble Pichia veronae and Pichia fabianii but differ from them by nine nucleotides (1.6%) in D1/D2 sequences. The two strains are described as Candida pattaniensis sp. nov. The remaining strain, ST-37, is related to Pichia americana and Pichia bimundalis but differs by six(1.1%) and seven (1.2%) nucleotides from these species, respectively. This strain is described as Candida nakhonratchasimensis sp. nov. PMID- 15747232 TI - Polyamine analysis for chemotaxonomy of thermophilic eubacteria: Polyamine distribution profiles within the orders Aquificales, Thermotogales, Thermodesulfobacteriales, Thermales, Thermoanaerobacteriales, Clostridiales and Bacillales. AB - Cellular polyamines of 45 thermophilic and 8 related mesophilic eubacteria were investigated by HPLC and GC analyses for the thermophilic and chemotaxonomic significance of polyamine distribution profiles. Spermidine and a quaternary branched penta-amine, N4-bis(aminopropyl)norspermidine, were the major polyamine in Thermocrinis, Hydrogenobacter, Hydrogenobaculum, Aquifex, Persephonella, Sulfurihydrogenibium, Hydrogenothermus, Balnearium and Thermovibrio, located in the order Aquificales. Thermodesulfobacterium and Thermodesulfatator belonging to the order Thermodesulfobacteriales contained another quaternary penta-amine, N4 bis(aminopropyl)spermidine. In the order Thermotogales, Thermotoga contained spermidine, norspermidine, caldopentamine and homocaldopentamine. The latter two linear penta-amines were not found in Marinitoga and Petrotoga. In the order Thermales, Thermus and Marinithermus contained homospermidine, norspermine and the linear penta-amines. Meiothermus lacked penta-amines. Vulcanithermus contained linear penta-amines and hexa-amines but not homospermidine. Oceanithermus contained spermine alone. Within the order Thermoanaerobacteriales, the two quaternary branched penta-amines were found in Thermanaeromonas and Thermoanaerobacter. Caldanaerobacter contained N4-bis(aminopropyl)spermidine. Thermoanaerobacterium lacked penta-amines. Thermaerobacter of the order Clostridiales contained N4-bis(aminopropyl)spermidine and agmatine. Thermosyntropha, Thermanaerovibrio, Thermobrachium ( the order Clostridiales), Sulfobacillus, Alicyclobacillus, Anoxybacillus, Ureibacillus, Thermicanus ( the order Bacillales), Desulfotomaculum, Desulfitobacterium and Pelotomaculum (the family Peptococcaceae) ubiquitously contained spermine. Some thermophiles of Bacillales added linear and branched penta-amines. PMID- 15747233 TI - Cellular polyamine profile of the phyla Dinophyta, Apicomplexa, Ciliophora, Euglenozoa, Cercozoa and Heterokonta. PMID- 15747234 TI - New perspectives on the persistent scourge of foodborne disease. PMID- 15747235 TI - Relationship between age and the risk of surgical site infection: a contemporary reexamination of a classic risk factor. PMID- 15747236 TI - In search of a vaccine for respiratory syncytial virus: the saga continues. PMID- 15747237 TI - Antimicrobial-resistant and extraintestinal pathogenic Escherichia coli in retail foods. AB - BACKGROUND: Extraintestinal Escherichia coli infections are associated with specialized extraintestinal pathogenic E. coli (ExPEC) strains and, increasingly, with antimicrobial resistance. The food supply may disseminate ExPEC and antimicrobial-resistant E. coli. METHODS: In a prospective survey of 1648 diverse food items from 10 retail markets in the Minneapolis-St. Paul area during 2001 2003, selective cultures and disk-diffusion assays for the isolation and characterization of antimicrobial-resistant E. coli and polymerase chain reaction based assays and O serotyping to define ExPEC-associated traits were performed. RESULTS: E. coli contamination exhibited a prevalence gradient from miscellaneous foods (9%), through beef or pork (69%), to poultry (92%; P<.001). Among E. coli positive samples, similar prevalence gradients were detected for antimicrobial resistance (27%, 85%, and 94% of samples, respectively; P<.001) and ExPEC contamination (4%, 19%, and 46%, respectively; P<.001). By multivariate analysis, beef or pork and poultry from natural-food stores exhibited reduced risks of E. coli contamination and antimicrobial resistance. Indirect evidence suggested on farm selection of resistance. Four food-source ExPEC isolates (from pea pods, turkey parts, ground pork, and vegetable dip) closely resembled selected human clinical isolates by O antigen and genomic profile. CONCLUSIONS: Retail foods may be an important vehicle for community-wide dissemination of antimicrobial resistant E. coli and ExPEC, which may represent a newly recognized group of medically significant foodborne pathogens. PMID- 15747238 TI - Adverse health events associated with antimicrobial drug resistance in Campylobacter species: a registry-based cohort study. AB - BACKGROUND: Resistance to clinically important antimicrobial agents, particularly fluoroquinolones and macrolides, is increasing among Campylobacter isolates, but few studies have explored the human health consequences of such resistance. METHODS: In a registry-based cohort study, we determined the risk of invasive illness and death associated with infection with quinolone- and erythromycin resistant Campylobacter strains, while adjusting for comorbidity. We linked data from the Danish Surveillance Registry for Enteric Pathogens with data from the Civil Registration System and National Health Registries. RESULTS: Of 3471 patients with Campylobacter infection, 22 (0.63%) had an adverse event, defined as invasive illness or death, within 90 days of the date of receipt of samples. Patients infected with quinolone-resistant Campylobacter strains had a 6-fold increased risk of an adverse event within 30 days of the date of receipt of samples, compared with patients infected with quinolone- and erythromycin susceptible Campylobacter strains (adjusted odds ratio [AOR], 6.17 [95% confidence interval {CI}, 1.62-23.47]). However, infection with erythromycin resistant strains was associated with a >5-fold risk of an adverse event within 90 days of the date of receipt of samples (AOR, 5.51 [95% CI, 1.19-25.50]). CONCLUSIONS: The present study provides evidence of the human health consequences of resistance to clinically important agents among Campylobacter infections and the need for increased efforts to mitigate such resistance. PMID- 15747239 TI - The effect of increasing age on the risk of surgical site infection. AB - BACKGROUND: An increasing number of older persons undergo surgery, but the relationship between increasing age and risk of surgical site infection (SSI) has not been established. The objective of the present study was to determine the relationship between increasing age and risk of SSI. METHODS: The present cohort study included patients who underwent surgery between February 1991 and July 2002. Patients >17 years of age were divided randomly into derivation and validation cohorts. The study was conducted at 11 hospitals. SSIs were prospectively identified by use of Centers for Disease Control and Prevention criteria. RESULTS: The study included 144,485 consecutive surgical patients and 1684 SSIs (rate of SSI, 1.2%). There were 72,139 procedures and 873 SSIs in the derivation cohort. Adjusted analyses revealed a significant relationship between age and risk of SSI (P=.006). Risk of SSI increased by 1.1%/year between ages 17 and 65 years (P=.002). At age >/=65 years, risk of SSI decreased by 1.2% for each additional year (P=.008). There were 72,334 procedures and 811 SSIs in the validation cohort. The relationship between age and risk of SSI was similar in the validation cohort. CONCLUSIONS: Increasing age independently predicted an increased risk of SSI until age 65 years. At ages >/=65 years, increasing age independently predicted a decreased risk of SSI. PMID- 15747240 TI - Administration of protein-conjugate pneumococcal vaccine to patients who have invasive disease after splenectomy despite their having received 23-valent pneumococcal polysaccharide vaccine. AB - Patients who undergo splenectomy are at greatly increased risk for overwhelming pneumococcal bacteremia and death. Twenty-three-valent pneumococcal polysaccharide vaccine (PPV-23), which contains capsular polysaccharides (PSs) from 23 common serotypes of Streptococcus pneumoniae, is strongly recommended for such patients. The capacity to respond to PPV-23 by producing immunoglobulin (Ig) G is genetically regulated. Some proportion of adults do not respond and, despite postsplenectomy administration of PPV-23, may remain susceptible to recurrent pneumococcal sepsis. Here, we describe 2 patients who had recurring pneumococcal bacteremia after undergoing splenectomy despite having received numerous doses of PPV-23. Heptavalent protein-conjugate pneumococcal vaccine (PCPV-7) was then administered, and it induced high levels of IgG to all 7 PSs; in one of the patients, functional activity against 5 of the 7 PSs was demonstrable, both in vitro and in vivo. Recurrent pneumococcal bacteremia in patients who have undergone splenectomy may indicate a genetically regulated failure to respond to PPV-23; PCPV-7 may stimulate production of IgG to PSs in such patients. PMID- 15747241 TI - Host susceptibility and clinical outcomes in toll-like receptor 5-deficient patients with typhoid fever in Vietnam. AB - Toll-like receptor 5 (TLR5) mediates innate immune responses to bacterial pathogens by binding to flagellin. A polymorphism in the TLR5 gene introduces a premature stop codon (TLR5(392STOP)) that is associated with susceptibility to legionnaires disease. Here we investigated whether TLR5(392STOP) was associated with typhoid fever. The frequency of TLR5(392STOP) was not significantly different in 565 patients with typhoid fever and 281 ethnically matched control subjects. Furthermore, TLR5 deficiency had no measurable effect on a number of clinical parameters associated with typhoid fever, including fever clearance time, pathogen burden, disease severity, or age at acquisition of disease. TLR5 may not play an important role in TLR-stimulated innate immune responses to human infection with Salmonella enterica serovar Typhi. Initiation of these responses may rely on other TLRs that recognize different bacterial ligands. PMID- 15747242 TI - Zinc inhibits cholera toxin-induced, but not Escherichia coli heat-stable enterotoxin-induced, ion secretion in human enterocytes. AB - BACKGROUND: Because zinc deficiency in malnourished children is associated with severe diarrhea, use of zinc supplementation has been proposed as an adjunct to oral rehydration. However, the effects of zinc on enterocyte ion transport are largely unknown. The objective of the present study was to investigate the effects of zinc on transepithelial ion transport under basal conditions and under conditions of enterotoxin-induced ion secretion. METHODS: Ion transport was investigated by monitoring electrical parameters in human intestinal Caco-2 cells that were mounted in Ussing chambers and exposed to increasing concentrations of zinc, both in the absence and presence of either cholera toxin (CT) or Escherichia coli heat-stable enterotoxin (ST). Intracellular cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) concentrations were also determined. RESULTS: The addition of zinc to the luminal or basolateral side of enterocytes induced a chloride-dependent, dose-related decrease in short circuit current, indicating ion absorption. It also resulted in a substantial reduction in CT-induced ion secretion and in cAMP concentration. E. coli ST induced ion secretion and cGMP concentration were not affected. Ion absorption peaked at 35 mu mol/L zinc, whereas excess zinc load induced active ion secretion. CONCLUSIONS: By causing a decrease in cAMP concentration, zinc directly promotes ion absorption and substantially reduces CT-induced, but not E. coli ST-induced, ion secretion. PMID- 15747243 TI - Escherichia coli strains belonging to phylogenetic group B2 have superior capacity to persist in the intestinal microflora of infants. AB - Escherichia coli strains segregate into 4 phylogenetic groups, designated "A," "B1," "B2," and "D." Pathogenic strains belong to group B2 and, to a lesser extent, group D, which more frequently carry virulence-factor genes than do group A strains and group B1 strains. This study investigated whether the capacity of E. coli to persist in the human intestine is related to its phylogenetic type. Resident (n=58) and transient (n=19) commensal E. coli strains isolated during a longitudinal study of 70 Swedish infants and previously tested for virulence factor-gene carriage were tested for phylogenetic type. Of the strains resident in the intestinal microflora, 60% belonged to group B2, compared with only 21% of the transient strains (P=.004). In logistic regression, group B2 type predicted persistence in the intestinal microflora, independent of carriage of all investigated virulence-factor genes, including genes for P fimbriae (P=.03). Thus, group B2 strains appear to possess yet unidentified traits that enhance their survival in the human intestine. PMID- 15747244 TI - Human leukocyte antigen and cytokine gene variants as predictors of recurrent Chlamydia trachomatis infection in high-risk adolescents. AB - Antigen presentation and immune activation are essential to the effective control of infectious diseases. In 485 North American adolescents at high risk for genital Chlamydia trachomatis infection, we found 2 human leukocyte antigen variants (DRB1*03-DQB1*04 and DQB1*06) to be associated with recurrent Chlamydia infection (adjusted relative odds [RO], >2.0; P<.01, for both variants). A G-C-C haplotype corresponding to variants at IL10 (encoding interleukin-10 [IL-10]) promoter positions -1082, -819, and -592 was underrepresented in individuals with recurrent infection (RO, 0.59; P=.046). These genetic associations were independent of nongenetic factors, including number of sex partners, race, sex, duration of follow-up, and human immunodeficiency virus type 1 seropositivity. Consistent with the observed IL10 association, cervical secretions in female adolescents without the IL10 G-C-C haplotype had elevated IL-10 concentrations after Chlamydia infection, which may reflect involvement of a Chlamydia-specific mechanism for genetically mediated, differential IL-10 expression in the genital tract. PMID- 15747245 TI - Identification of a recombinant live attenuated respiratory syncytial virus vaccine candidate that is highly attenuated in infants. AB - BACKGROUND: Recombination technology can be used to create live attenuated respiratory syncytial virus (RSV) vaccines that contain combinations of known attenuating mutations. METHODS: Two live attenuated, recombinantly derived RSV vaccine candidates, rA2cp248/404 Delta SH and rA2cp248/404/1030 Delta SH, were evaluated in 31 adults and in 95 children >/=6 months old. rA2cp248/404/1030 Delta SH was subsequently evaluated in 44 infants 1-2 months old. These vaccine candidates share 4 attenuating genetic elements and differ only in a missense mutation (1030) in the polymerase gene. RESULTS: Both vaccines were highly attenuated in adults and RSV-seropositive children and were well tolerated and immunogenic in RSV-seronegative children. Compared with that of rA2cp248/404 Delta SH, replication of rA2cp248/404/1030 Delta SH was restricted in RSV seronegative children (mean peak titer, 10(4.3) vs. 10(2.5) plaque-forming units [pfu]/mL), indicating that the 1030 mutation had a potent attenuating effect. Although rA2cp248/404/1030 Delta SH was well tolerated in infants, only 44% of infants who received two 10(5.3)-pfu doses of vaccine had detectable antibody responses. However, replication after administration of the second dose was highly restricted, indicating that protective immunity was induced. At least 4 of 5 attenuating genetic elements were retained in recovered vaccine viruses. CONCLUSIONS: rA2cp248/404/1030 Delta SH is the first RSV vaccine candidate to be sufficiently attenuated in young infants. Additional studies are needed to determine whether rA2cp248/404/1030 Delta SH can induce protective immunity against wild-type RSV. PMID- 15747246 TI - Mobilization of plasmacytoid and myeloid dendritic cells to mucosal sites in children with respiratory syncytial virus and other viral respiratory infections. AB - BACKGROUND: Respiratory syncytial virus (RSV) is the principal etiologic agent of bronchiolitis and viral pneumonia in infants and young children. Yet, many aspects of its immunopathogenesis are not well understood. METHODS: We analyzed the immune cells that are mobilized by RSV and other respiratory viruses, by studying nasal wash samples from children hospitalized with acute viral respiratory infections. RESULTS: RSV mobilizes virtually all blood immune cells, including myeloid dendritic cells (DCs) and plasmacytoid DCs (pDCs), to the nasal mucosa. DCs were also mobilized to the nasal mucosa of children with other viral respiratory infections. The increased number of pDCs in the nasal compartment significantly correlates with RSV load (P=.022), and it is associated with a significant decrease in the number of pDCs in the blood (P=.007). The influx of DCs in the nasal mucosa is not transient, as even higher numbers of both DC subsets were found in respiratory secretions weeks after the acute symptoms of RSV infection had resolved. Immunochemistry analysis of respiratory samples has demonstrated the presence of the RSV fusion protein within HLA-DR-positive cells. CONCLUSION: Infection with RSV and other respiratory viruses mobilizes DCs to the site of viral entry. PMID- 15747247 TI - A bovine parainfluenza virus type 3 vaccine is safe and immunogenic in early infancy. AB - BACKGROUND: A phase 2 trial was conducted to assess in young infants the safety, tolerability, infectivity, and immunogenicity of multiple doses of an intranasal vaccine using bovine parainfluenza virus type 3 (bPIV3). METHODS: One hundred ninety-two healthy 2-month-old infants were randomized 1 : 1 : 1 to receive 1x10(5) median tissue culture infective dose (TCID(50)) bPIV3 vaccine, 1x10(6) TCID(50) bPIV3 vaccine, or placebo at 2, 4, 6, and 12-15 months of age. Safety information was collected by use of diary sheets and telephone interviews. Nasal wash and serum specimens were collected for assessment of infectivity and immunogenicity. RESULTS: The safety profiles of both dosages of bPIV3 were similar to that of placebo, with the exception of fever with temperature of >/=38.1 degrees C after dose 2 only, occurring in 34% of the 1x10(5) TCID(50) group, 35% of the 1x10(6) TCID(50) group, and 12% of the placebo group (P<.01). No vaccine-related serious adverse events were reported. The cumulative vaccine infectivity (isolation of bPIV3 and/or bPIV3 seroconversion) after dose 3 was similar in the 2 vaccine groups (87% in the 1x10(5) TCID(50) group and 77% in the 1x10(6) TCID(50) group) (P=.46). Seroconversion rates after dose 3, assessed by means of hemagglutination inhibition assay, after adjustment for decrease in maternal antibody titers, were 67% in the 1x10(5) TCID(50) group, 57% in the 1x10(6) TCID(50) group, and 12% in the placebo group (P<.01). Isolation of bPIV3 was common after dose 1, dose 2, or dose 3, but only 1 of 51 participants in the vaccine groups had bPIV3 isolated after dose 4. CONCLUSIONS: Multiple doses of bPIV3 vaccine were well tolerated and immunogenic in young infants. PMID- 15747248 TI - The rat-based neurovirulence safety test for the assessment of mumps virus neurovirulence in humans: an international collaborative study. AB - Because of the highly neurotropic and neurovirulent properties of wild-type mumps viruses, most national regulatory organizations require neurovirulence testing of virus seeds used in the production of mumps vaccines. Such testing has historically been performed in monkeys; however, some data suggest that testing in monkeys does not necessarily discriminate among the relative neurovirulent risks of mumps virus strains. To address this problem, a collaborative study was initiated by the National Institute for Biological Standards and Control in the United Kingdom and the Food and Drug Administration in the United States, to test a novel rat-based mumps virus neurovirulence safety test. Results indicate that the assay correctly assesses the neurovirulence potential of mumps viruses in humans and is robust and reproducible. PMID- 15747249 TI - Effects of bacterial vaginosis and other genital infections on the natural history of human papillomavirus infection in HIV-1-infected and high-risk HIV-1 uninfected women. AB - BACKGROUND: Whether the natural history of human papillomavirus (HPV) infection is affected by bacterial vaginosis (BV) or Trichomonas vaginalis (TV) infection has not been adequately investigated in prospective studies. METHODS: Human immunodeficiency virus 1 (HIV-1)-infected (n=1763) and high-risk HIV-1-uninfected (n=493) women were assessed semiannually for BV (by Nugent's criteria), TV infection (by wet mount), type-specific HPV (by polymerase chain reaction with MY09/MY11/HMB01 HPV primers), and squamous intraepithelial lesions (SIL) (by cytological examination). Sexual history was obtained from patient report at each visit. Risk factors for prevalent and incident HPV infection and SIL were evaluated by use of multivariate models. RESULTS: BV was associated with both prevalent and incident HPV infection but not with duration of HPV infection or incidence of SIL. TV infection was associated with incident HPV infection and with decreased duration and lower prevalence of HPV infection. TV infection had no association with development of SIL. Effects of BV and TV infection were similar in HIV-1-infected and high-risk HIV-1-uninfected women. HIV-1 infection and low CD4(+) lymphocyte count were strongly associated with HPV infection and development of SIL. CONCLUSIONS: BV and TV infection may increase the risk of acquisition (or reactivation) of HPV infection, as is consistent with hypotheses that the local cervicovaginal milieu plays a role in susceptibility to HPV infection. The finding that BV did not affect persistence of HPV infection and that TV infection may shorten the duration of HPV infection helps explain the lack of effect that BV and TV infection have on development of SIL. PMID- 15747250 TI - The clonal expansion of human T lymphotropic virus type 1-infected T cells: a comparison between seroconverters and long-term carriers. AB - BACKGROUND: The clonal expansion of human T lymphotropic virus type 1 (HTLV-1) infected T cells is considered to be important for the maintenance of infection. However, the process by which the clonality of HTLV-1-infected T cells is established is not understood. METHODS: HTLV-1 clonality in 4 adult seroconverters was analyzed by inverse long polymerase chain reaction (PCR) followed by cloning of the PCR products and evaluation of restriction fragment length polymorphism. The results were compared with those for 8 long-term HTLV-1 carriers. RESULTS: The clonality of HTLV-1-infected T cells in the seroconverters arose stochastically and was variable 3-5 years after seroconversion. On the basis of the frequency with which clones of cells infected with unique HTLV-1 provirus integration sites appeared, it was clear that the seroconverters had a greater number of unique clones with fewer infected cells than did the long-term carriers. CONCLUSIONS: The clonality of the HTLV-1-infected T cells in the adult seroconverters, who had been newly infected via HTLV-1-carrier spouses, was more heterogeneous and less stable than that of the HTLV-1-infected T cells in long term carriers, who were more likely to have been infected during infancy. The mechanism for the selective maintenance of certain clones in asymptomatic HTLV-1 carriers likely plays a role in the initiation of leukemogenesis. PMID- 15747251 TI - Use of RNA interference to prevent lethal murine west nile virus infection. AB - West Nile virus causes fatal encephalitis in humans, mice, and other vertebrates. In the present study, we demonstrate that small interfering RNAs (siRNAs) inhibit West Nile virus replication in vitro. Moreover, the administration of siRNAs to mice by hydrodynamic injection 24 h before challenge with an intraperitoneal inoculum of West Nile virus reduced the viral load and afforded partial protection from lethal infection. These data show the efficacy of the prophylactic use of siRNAs against a viral infection in vivo and suggest new strategies to combat West Nile virus. PMID- 15747252 TI - T-1249 retains potent antiretroviral activity in patients who had experienced virological failure while on an enfuvirtide-containing treatment regimen. AB - BACKGROUND: T-1249 is a 39-amino acid synthetic peptide fusion inhibitor (FI) shown to preserve antiretroviral activity in vitro against human immunodeficiency virus (HIV) isolates that have decreased susceptibility to enfuvirtide (ENF). METHODS: A 10-day phase 1/2 study of the safety and antiretroviral activity of T 1249 was conducted in 53 HIV-1-infected adults with detectable viremia while on an ENF-containing treatment regimen. RESULTS: From FI-naive baseline levels, the geometric mean (GM) decrease in susceptibility to ENF was 116.3-fold, and the GM decrease in susceptibility to T-1249 was 2.0-fold. Patients continued to administer their failing treatment regimen but replaced ENF with T-1249 at a dose of 192 mg/day. T-1249 was generally well tolerated; injection site reactions, which were generally mild, were the most commonly reported adverse event (64% of patients). The median change from levels of HIV-1 RNA at baseline to levels on day 11 was -1.26 log(10) copies/mL (95% confidence interval, -1.40 to -1.09 log(10) copies/mL); on day 11, a decrease from baseline HIV-1 RNA levels of >/=1.0 log(10) copies/mL was seen in 73% of patients. Antiretroviral activity, as measured by levels of HIV-1 RNA, was not predicted by baseline susceptibility to T-1249 or to ENF; genotypic substitutions that emerged during T-1249 treatment were identified in virus from some patients. CONCLUSIONS: These results indicate that FIs constitute an expanding class of antiretroviral agents with the potential to be sequenced. PMID- 15747253 TI - Predictors of residual viremia in HIV-infected patients successfully treated with efavirenz and lamivudine plus either tenofovir or stavudine. AB - In human immunodeficiency virus (HIV)-infected patients successfully treated with highly active antiretroviral therapy (HAART), a low level of HIV RNA persists in plasma at steady state for years and varies among patients. To understand predictors of residual viremia, we measured HIV RNA levels <50 copies/mL in patients after 1 year of treatment with efavirenz and lamivudine plus either tenofovir disoproxil fumarate (n=55) or stavudine (n=45), by use of an HIV RNA assay with a limit of detection of 2.5 copies/mL. The mean posttreatment HIV RNA levels were 0.58 log(10) copies/mL (3.8 copies/mL) in the tenofovir arm and 0.61 log(10)copies/mL (4.1 copies/mL) in the stavudine arm (P=.24). Forty-seven percent of patients receiving tenofovir, compared with 29% of patients receiving stavudine, had undetectable residual viremia (P=.07). In multivariate analyses, we found that lower baseline HIV RNA levels in plasma, lower HIV DNA levels in peripheral blood mononuclear cells, and inclusion in the tenofovir arm each independently predicted undetectable residual viremia (P<.05). However, a level of residual viremia <50 copies/mL was not associated with CD4 cell count changes or risk of virologic rebound through 72 weeks of follow-up. PMID- 15747254 TI - Effect of HIV infection and antiretroviral therapy on hepatitis B virus (HBV) specific T cell responses in patients who have resolved HBV infection. AB - Coinfection with hepatitis B virus (HBV) is a common occurrence in human immunodeficiency virus (HIV)-positive patients and an increasing cause of morbidity and mortality. The CD8(+) T cell response is critical for long-term control of HBV in patients resolving acute infection. Here, we examine the effect of HIV on HBV-specific CD8(+) T cell responses in patients who have resolved HBV infection. A cross-sectional study showed a reduction in HBV-specific CD8(+) T cell responses in HIV-positive, HBV-immune patients, compared with those in HIV negative, HBV-immune patients. A longitudinal study of a subgroup of patients examined whether this attrition could be reversed by effective antiretroviral therapy. The introduction of highly active antiretroviral therapy (HAART) resulted in reconstitution of some HBV-specific CD4(+) and CD8(+) T cell responses, in association with restoration of CD4(+) T cell counts. These data provide a mechanism to account for the observed impairment of control of HBV infection in the setting of HIV infection and support the ability of HAART to reconstitute functionally active T cell responses. PMID- 15747255 TI - Interferon- gamma and granulocyte-macrophage colony-stimulating factor augment the activity of polymorphonuclear leukocytes against medically important zygomycetes. AB - Zygomycetes cause serious invasive infections, predominantly in immunocompromised and diabetic patients with poor prognoses and limited therapeutic options. We compared the antifungal function of human polymorphonuclear leukocytes (PMNLs) against hyphae of Rhizopus oryzae and R. microsporus, the most frequently isolated zygomycetes, with that against the less frequently isolated Absidia corymbifera. We then evaluated the effects of interferon (IFN)- gamma and granulocyte-macrophage colony-stimulating factor (GM-CSF), alone or combined, on PMNL antifungal function against these zygomycetes. Both PMNL oxidative burst in response to hyphae and PMNL-induced hyphal damage were significantly lower in response to Rhizopus species than in response to A. corymbifera. Incubation of PMNLs with IFN- gamma and GM-CSF alone or combined for 22 h increased the PMNL induced hyphal damage of all 3 species. The treatment of PMNLs with the combination of IFN- gamma and GM-CSF significantly increased the release of tumor necrosis factor- alpha in response to R. microsporus and A. corymbifera hyphae. IFN- gamma significantly reduced interleukin-8 release in response to all zygomycetes. Although Rhizopus species demonstrate a decreased susceptibility to the antifungal activity of human PMNLs, in comparison with A. corymbifera, IFN- gamma and GM-CSF augment the hyphal damage of all 3 zygomycetes, suggesting a role for IFN- gamma and GM-CSF in the management of invasive zygomycosis. PMID- 15747256 TI - Toll-deficient Drosophila flies as a fast, high-throughput model for the study of antifungal drug efficacy against invasive aspergillosis and Aspergillus virulence. AB - Invasive aspergillosis (IA) is the most important opportunistic mycosis in immunosuppressed patients. The lack of a sufficient number of effective antifungals and our incomplete understanding of the pathogenesis of IA contribute to its overall unfavorable prognosis. Studies of drug efficacy against IA and Aspergillus virulence rely on conventional animal models that are laborious and use limited numbers of animals; alternative, less cumbersome in vivo models are desirable. Using different inoculation models of IA, we found that Toll-deficient Drosophila flies exposed to voriconazole (VRC), the preferred drug for the treatment of IA in humans, had significantly better survival rates and lower tissue fungal burdens than did those not exposed to VRC. Furthermore, Toll deficient Drosophila flies infected with an alb1-deleted hypovirulent Aspergillus mutant had significantly better survival rates than did those infected with a wild-type Aspergillus strain. Therefore, the Drosophila fly is a fast, high throughput in vivo model for the study of drug efficacy against IA and Aspergillus virulence. PMID- 15747257 TI - In vivo transcriptome of Plasmodium falciparum reveals overexpression of transcripts that encode surface proteins. AB - Infections with the human parasite Plasmodium falciparum continue to present a great challenge to global health. Fundamental questions regarding the molecular basis of virulence and immune evasion in P. falciparum have been only partially answered. Because of the parasite's intracellular location and complex life cycle, standard genetic approaches to the study of the pathogenesis of malaria have been limited. The present study presents a novel approach to the identification of the biological processes involved in host-pathogen interactions, one that is based on the analysis of in vivo P. falciparum transcripts. We demonstrate that a sufficient quantity of P. falciparum RNA transcripts can be derived from a small blood sample from infected patients for whole-genome microarray analysis. Overall, excellent correlation was observed between the transcriptomes derived from in vivo samples and in vitro samples with ring-stage P. falciparum 3D7 reference strain. However, gene families that encode surface proteins are overexpressed in vivo. Moreover, this analysis has identified a new family of hypothetical genes that may encode surface variant antigens. Comparative studies of the transcriptomes derived from in vivo samples and in vitro 3D7 samples may identify important strategies used by the pathogen for survival in the human host and highlight, for vaccine development, new candidate antigens that were not previously identified through the use of in vitro cultures. PMID- 15747259 TI - No convincing evidence of linkage for restless legs syndrome on chromosome 9p. PMID- 15747261 TI - Competing risks: preoperative myocardial infarction or postoperative bleeding? PMID- 15747258 TI - A high-resolution linkage-disequilibrium map of the human major histocompatibility complex and first generation of tag single-nucleotide polymorphisms. AB - Autoimmune, inflammatory, and infectious diseases present a major burden to human health and are frequently associated with loci in the human major histocompatibility complex (MHC). Here, we report a high-resolution (1.9 kb) linkage-disequilibrium (LD) map of a 4.46-Mb fragment containing the MHC in U.S. pedigrees with northern and western European ancestry collected by the Centre d'Etude du Polymorphisme Humain (CEPH) and the first generation of haplotype tag single-nucleotide polymorphisms (tagSNPs) that provide up to a fivefold increase in genotyping efficiency for all future MHC-linked disease-association studies. The data confirm previously identified recombination hotspots in the class II region and allow the prediction of numerous novel hotspots in the class I and class III regions. The region of longest LD maps outside the classic MHC to the extended class I region spanning the MHC-linked olfactory-receptor gene cluster. The extended haplotype homozygosity analysis for recent positive selection shows that all 14 outlying haplotype variants map to a single extended haplotype, which most commonly bears HLA-DRB1*1501. The SNP data, haplotype blocks, and tagSNPs analysis reported here have been entered into a multidimensional Web-based database (GLOVAR), where they can be accessed and viewed in the context of relevant genome annotation. This LD map allowed us to give coordinates for the extremely variable LD structure underlying the MHC. PMID- 15747262 TI - Preoperative use of enoxaparin increases the risk of postoperative bleeding and re-exploration in cardiac surgery patients. AB - OBJECTIVE: The purpose of this study was to investigate if the preoperative use of new platelet inhibitors and low-molecular-weight heparins may contribute to bleeding after cardiac surgery. DESIGN: Retrospective data review. SETTING: University teaching hospital. PARTICIPANTS: One hundred eleven patients divided in 5 groups. INTERVENTIONS: Patients were grouped according to preoperative antithrombotic regimen: group 1, control, no agents (n=55); group 2, clopidogrel (n=9); group 3, enoxaparin (n=17); group 4, any GP IIb/IIIa inhibitor (n=14); and group 5, any drug combination (n=15). Data included cumulative mediastinal chest tube drainage, allogeneic blood transfusions, total blood donor exposures, and re exploration. MEASUREMENTS AND MAIN RESULTS: Use of any drug (groups 2-5) resulted in greater total blood transfusions and donor exposure (p=0.0003) than control, especially red cells (p=0.002) and platelets (p=0.006). A greater percentage of patients on enoxaparin required mediastinal re-exploration for nonsurgical bleeding versus control (3/17 v 0/55, p=0.001). The use of enoxaparin was associated with significantly higher chest tube output after the first 24 hours postoperatively (p=0.048). CONCLUSION: Newer antithrombotic agents were associated with greater transfusion rates and total donor exposures. Enoxaparin use was associated with greater overall blood loss and with higher incidence of mediastinal re-exploration. The relative risk-benefit ratio of reduced periprocedure morbidity versus increased bleeding complications has yet to be determined. PMID- 15747263 TI - Hypercoagulability affecting early vein graft patency does not exist after off pump coronary artery bypass. AB - OBJECTIVE: Hypercoagulability may compromise the patency of bypass grafts. The authors hypothesized that perioperative in vitro platelet responses to varying agonists (eg, thrombin, platelet activating factor, collagen, adenosine diphosphate) correlate with early graft thrombosis after off-pump coronary artery bypass grafting (OPCAB). DESIGN AND PARTICIPANTS: Prospective study of 78 OPCAB patients with 151 venous bypass grafts treated with perioperative aspirin and intraoperative heparin (250 U/kg). SETTING: Tertiary, academic medical center. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Hypercoagulability, defined by TEG (maximum amplitude [MA]>70 mm), whole-blood aggregometry (>15 ohms after 5 mcl/mL collagen) or hemoSTATUS (Ch5CR>0.5), was serially assessed around OPCAB. An immediate decline in platelet function after surgery and on postoperative day 1 returned to normal by postoperative day 3 in most patients. Graft blood flow was analyzed intraoperatively, and vein biopsies were analyzed for endothelial disruption. Graft patency was assessed by multichannel computed tomography coronary angiography on postoperative day 5. No differences in any of the platelet function assays were noted for the 8 patients with graft thrombosis (n=8 grafts) versus the 68 patients with all patent grafts (n=129 grafts). Ten patients developed a rise in platelet function postoperatively >1 SD above baseline; only 1 developed graft thrombosis (p=not significant v patients with normal platelet function). CONCLUSIONS: OPCAB is not associated with a significant activation in postoperative platelet function. This study suggests that if hypercoagulability exists after OPCAB, it is not involved in the pathogenesis of arterial thrombotic events such as early bypass graft failure. PMID- 15747265 TI - Resource utilization in on- and off-pump coronary artery surgery: factors influencing postoperative length of stay--an experience of 1,746 consecutive patients undergoing fast-track cardiac anesthesia. AB - OBJECTIVE: The purpose of the present investigation was to examine factors influencing resource utilization in patients undergoing on-pump coronary artery bypass graft and off-pump coronary artery bypass (OPCAB) graft surgery at a major university hospital. The resources examined were time to extubation, packed red blood cell (PRBC) transfusion, intensive care length of stay (ICULOS), preoperative and postoperative length of stay (PLOS), and total length of stay (LOS). DESIGN: Observational study of consecutive patients undergoing on- and off pump coronary artery bypass surgery. SETTING: Tertiary care cardiac referral center. PARTICIPANTS: One thousand seven hundred forty-six consecutive male and female patients undergoing primary coronary artery bypass graft (CABG) surgery over a period of 3 years (1999-2001). Eight hundred eighty-one patients underwent CABG with pump, and 865 patients underwent off-pump coronary artery bypass (OPCAB) surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The mean time to extubation after surgery was 7.4 hours for on-pump patients and 5.8 hours for the OPCAB group (p72 hours to postoperative tracheal extubation compared with 1.5% in the OPCAB group (p=0.041). Hospital mortality was 2.7% for the on-pump group and 1.0% for the OPCAB group (p=0.010). CONCLUSION: The authors found that patients undergoing on pump CABG have significantly longer time to tracheal extubation, increased blood use, longer ICULOS, PLOS, and total LOS and higher in-hospital mortality, which would translate into significant differences in the expenses associated with these 2 surgical approaches to coronary surgery. PMID- 15747264 TI - Perturbations in laboratory values after coronary artery bypass graft surgery with cardiopulmonary bypass. AB - OBJECTIVE: The purpose of this study was to describe the sequential changes in commonly obtained laboratory values after coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). METHODS: The authors examined laboratory data from 375 patients who underwent uncomplicated CABG with CPB in a multicenter clinical trial of a medication for postoperative pain. Data were obtained preoperatively, at the time of postoperative extubation, and at 4 subsequent intervals ending 14 days after extubation. Data obtained before study drug administration are reported for all patients; thereafter, only data from placebo patients without perioperative complications (n=123) are reported. RESULTS: Mean postoperative coagulation values remained within their reference ranges at the time of extubation. However, platelet counts increased to a peak value well above the reference range by the end of the study. Postoperative white blood cell counts rose above the reference range, mainly because of increased neutrophils. Serum chemistries were also altered; most patients showed a persistent postoperative hyperglycemia. Creatine kinase levels rose to nearly 4 times the upper limit of the reference range in the early postoperative period. Lactate dehydrogenase, serum aspartate aminotransferase, and alanine aminotransferase levels also increased above the reference range. Total protein and albumin values were below the reference range throughout the postoperative period. CONCLUSIONS: Laboratory values for hematology, blood coagulation, and serum chemistry change substantially after uncomplicated CABG with CPB. Recognition of these changes will facilitate the conduct of clinical research and may prevent inappropriate treatment based on abnormal laboratory findings that have no clinical significance. PMID- 15747266 TI - Comparison of three anesthetic techniques for off-pump coronary artery bypass grafting: general anesthesia, combined general and high thoracic epidural anesthesia, or high thoracic epidural anesthesia alone. AB - OBJECTIVE: This study compared general anesthesia (GA), combined GA plus thoracic epidural anesthesia (TEA), and TEA alone in patients scheduled for off-pump coronary artery bypass grafting. DESIGN: Prospective, nonrandomized clinical study SETTING: University hospital. PARTICIPANTS: Ninety consenting patients undergoing beating-heart coronary artery revascularization with comparable coronary status and left ventricular function. INTERVENTIONS: GA (n=30) was conducted with propofol, remifentanil, and cisatracurium or combined with TEA (GA+TEA, n=30) or TEA as the sole anesthetic with ropivacaine plus sufentanil (TEA, n=30). MEASUREMENTS AND MAIN RESULTS: Groups were comparable regarding the surgical approaches and the number of anastomoses. Four patients (GA, n=2; GA+TEA, n=2) who required unplanned cardiopulmonary bypass, and 4 patients in the TEA group who underwent unexpected intubation because of pneumothorax (n=2), phrenic nerve palsy, or incomplete analgesia were excluded from further analysis. Intraoperative heart rate decreased significantly with both GA+TEA and TEA. None of the patients with TEA alone was admitted to the intensive care unit, they all were monitored on average for 6 hours postoperatively in the intermediate care unit and allowed to eat and drink as desired on admission. Postoperative pain scores were lower in both groups with TEA. There were no differences among groups in patients overall satisfaction. CONCLUSION: Based on the authors data, all anesthetic techniques were equally safe from the clinicians standpoint. However, GA+TEA appeared to be most comprehensive, allowing for revascularization of any coronary artery, providing good hemodynamic stability and reliable postoperative pain relief. Nonetheless, the actual and potential risks of TEA during cardiac surgery should not be underestimated. PMID- 15747267 TI - The effects of intrathecal morphine on patients undergoing minimally invasive direct coronary artery bypass surgery. AB - OBJECTIVE: To evaluate the effects of intrathecal morphine (ITM) on the perioperative use of opiates and the fast-track pathway in patients undergoing minimally invasive direct coronary artery bypass grafting (MIDCAB). DESIGN: Retrospective cohort study. SETTINGS: University teaching hospital. PARTICIPANTS: Twenty-two elective consecutive patients who underwent MIDCAB surgery. INTERVENTIONS: Eleven patients received an intrathecal injection of morphine, 7 microg/kg (ITM group, n=11), and the control group (n=11) received standard anesthesia care. MEASUREMENTS AND MAIN RESULTS: The intraoperative dose of fentanyl was 0.55+/-0.2 mg in the ITM group and 1.1+/-0.2 mg in the control group (p<0.0001). Four patients in the ITM group and none in the control group were extubated in the operating room. Extubation time was 3.2+/-4.7 hours in the ITM group and 6.3+/-3.4 hours in the control group (p=0.016). Morphine requirements in 24 hours after surgery were 2.0+/-3.5 mg in the ITM and 16.1+/-4.8 mg in the control group (p<0.0001). There were no differences in ischemic time, intraoperative fluid requirements, postoperative blood loss, and duration of surgical procedure. Hospital length of stay was shorter in the ITM group compared with the control group (3.9+/-1.7 days v 4.9+/-1.4 days, respectively, p=0.047). CONCLUSIONS: ITM has to be considered in MIDCAB surgery as an effective component of the perioperative analgesia. The safety and effects of ITM in the patients recovery after MIDCAB surgery should be evaluated in further prospective studies. PMID- 15747268 TI - Thoracic epidural anesthesia in cardiac surgical patients: a prospective audit of 2,113 cases. AB - OBJECTIVE: The purpose of this study was to present an audit report of thoracic epidural anesthesia without permanent neurologic deficits in more than 2,000 patients undergoing cardiac surgery. DESIGN: A prospective audit of cases conducted over a 13-year period. SETTING: Tertiary referral heart hospital. PARTICIPANTS: Two thousand one hundred thirteen patients over a period of 13 years. INTERVENTIONS: Epidural catheters were inserted at the C7 to T3 intervertebral space on the day before the operation in all patients; cardiac surgery was performed with or without cardiopulmonary bypass. MEASUREMENTS AND RESULTS: The authors did not encounter any permanent neurologic deficits in their series. The authors encountered 18 (0.85%) primary dural punctures and 4 cases (0.18%) of temporary neurologic deficits. CONCLUSION: This series adds to the worldwide experience of the use of epidural analgesia concomitantly with anticoagulation in cardiac surgery without serious complications. PMID- 15747269 TI - Fast-track coronary artery bypass grafting surgery under general anesthesia with remifentanil and spinal analgesia with morphine and clonidine. AB - OBJECTIVE: Effective postoperative analgesia is a critical part of fast-track cardiac surgery. This study compared the postoperative analgesic effect of fast track anesthesia with remifentanil and spinal morphine and clonidine with that of sufentanil anesthesia followed by patient-controlled administration of intravenous morphine. DESIGN: Prospective, blinded, randomized study. SETTING: Single private institution. PARTICIPANTS: Forty patients selected for coronary artery bypass graft surgery allocated randomly into 2 groups. INTERVENTIONS: General anesthesia was performed with etomidate, isoflurane, cisatracurium, and either remifentanil (0.10-0.25 microg/kg/min) or sufentanil (up to 3.5 microg/kg). In the remifentanil group, patients received spinal morphine (4 microg/kg) and clonidine (1 microg/kg) before induction. Postoperatively, patients in both groups were connected to an intravenous patient-controlled analgesia (PCA) morphine pump that delivered a 1-g bolus with a 7-minute lockout interval. MEASUREMENTS AND MAIN RESULTS: Patients were evaluated for pain on a visual analog scale (VAS), at rest and on deep breathing, and for intravenous PCA morphine consumption during 24 hours. The intravenous PCA morphine 24-hour cumulative dose was lower in the fast-track than in the control group (15.8+/ 12.6 v 32.7+/-22.3 mg, p<0.05). Before extubation, VAS scores were higher in the fast-track group, but after they were lower both at rest and during deep breathing. Extubation delay was shorter in the fast-track group (156.5+/-46.1 v 272+/-116.4 minutes, p<0.05). CONCLUSION: The combination of anesthesia with remifentanil and spinal analgesia with morphine and clonidine produces effective analgesia after coronary artery surgery and a rapid extubation time. PMID- 15747270 TI - Comparison of phenoxybenzamine to sodium nitroprusside in infants undergoing surgery. AB - OBJECTIVES: The purpose of this study was to compare the effects of a direct acting arterial dilator, sodium nitroprusside, to an alpha-adrenergic receptor blocker, phenoxybenzamine, in infants with congenital heart defects undergoing cardiac repairs on cardiopulmonary bypass. DESIGN: A prospective, multicenter, observational study. SETTING: Tertiary care center. PARTICIPANTS: Sixty infants scheduled for elective congenital cardiac surgery repair requiring cardiopulmonary bypass. INTERVENTIONS: Patients received either sodium nitroprusside 2 to 5 microg/kg/min infusion intraoperatively and in the intensive care unit (n=30 patients) or received phenoxybenzamine 1 mg/kg slowly intravenously at the onset of cardiopulmonary bypass (n=30 patients). MEASUREMENT AND MAIN RESULTS: Despite similar mean arterial pressures during cardiopulmonary bypass in both groups, infants who received phenoxybenzamine had a significantly higher flow compared with those who received sodium nitroprusside (180+/-4.8 v 73+/-5.12 mL/kg/min, p<0.0001). Base deficit was significantly larger in the sodium nitroprusside group compared with the phenoxybenzamine group intraoperatively and postoperatively (3.4+/-0.5 v 1.3+/-0.5 mEq/L, p<0.05). The core-to-peripheral temperature gradient was significantly larger in the sodium nitroprusside group compared with the phenoxybenzamine group intra- and postoperatively at all points studied. In the intensive care unit, the left atrial pressure was significantly higher in the sodium nitroprusside group compared with the phenoxybenzamine group (9+/-0.4 v 7+/-0.4 mmHg, p or =4.0 g/dL) and had a lower resistance to recombinant human erythropoietin (rHuEPO) therapy (by reducing the rHuEPO doses to reach the maintenance target values of hemoglobin (Hb) although both therapies resulted in equal depurative efficiency, and improved patient rehabilitation. CONCLUSIONS: This preliminary research, which requires further confirmation, demonstrates that HFR seems to provide PFD with other positive benefits and offers the uremic patient a better life style. PMID- 15747293 TI - [The IL-6 soluble receptors in hemodialyzed patients]. AB - Pro-inflammatory cytokines, in addition to their role in host defence, can be considered a disease mediator; therefore, a reduction in cytokine synthesis or its effects is becoming a target of many diseases. Interleukin-6 (IL-6) is a pro inflammatory cytokine that could play a role in several clinical problems related to dialysis treatment. Biological activities of IL-6 could be modulated by two soluble circulating receptors, namely sIL-6R and sgp130. sIL-6R can enhance the inflammatory effects of IL-6 and; therefore, is an "agonistically" acting molecule. On the contrary, sgp130 efficiently binds the IL-6/sIL-6R complex with "antagonistic" effects. In this study we evaluated sgp130 release by peripheral blood mononuclear cells (PBMC) harvested from 10 healthy controls (CON) and 11 end-stage renal disease (ESRD) patients undergoing renal dialysis therapy RDT) with cellulosic hemophan membrane (HD). We also evaluated gp130 gene expression by reverse transcriptase polymerase chain reaction (RT-PCR). gp130 is the membrane bound receptor of IL-6 that could be proteolytically cleaved to generate soluble sgp130. Our results demonstrated that HD. at basal conditions, showed a higher release of sgp130 as compared with CON. We also demonstrated by RT-PCR at basal conditions a higher gene expression of gp130 in HD, as compared with CON. These results took place in the absence of any mitogenic stimulation and suggest that in HD patients an inflammatory subclinical status increases sgp130 release. The results obtained after lipopolysaccharide (LPS) stimulation confirm the role of inflammation on the increased release of sgp130 in HD patients. PMID- 15747294 TI - [Pre- and post-dilution hemodiafiltration compared]. PMID- 15747295 TI - [The hemodiafiltration with infusion of acetate-free dialysis fluid can modify the inflammatory response in patients "high responders" to inflammatory stimuli?]. AB - PURPOSE: This study aimed to verify the effects of paired hemodiafiltration on line (PHF-AF) on inflammation in patients who were "high responders" to inflammatory stimuli: elevated C-reactive protein (CRP), genetic polymorphisms influencing a low transcription for interleukin-10 (IL-10) and a high transcription for IFN-gamma. METHODS: Ten patients selected as high responders for IFN-gamma and low responders for IL-10 were included in a crossover study to compare PHF-AF and standard bicarbonate hemodialysis (BHD). At study entry and before the start of each dialysis session the following examinations were performed: CRP, albumin, fibrinogen, ferritin, transferrin, prealbumin and serum levels of IL-6, IL-10, IFN-gamma, tumor necrosis factor-alpha (TNF-alpha). After the 1st and 3rd week of the study, the blood samples were also collected after the dialysis session. RESULTS: . There was a significant reduction in albumin and prealbumin in PHF-AF patients during the study; none of the other parameters were changed in both patient groups. CRP tended to be elevated after dialysis in both PHF-AF and BHD. While IL-6, IL-10 and IFN-gamma were unchanged during the dialysis session, there was a significant variation in TNF-alpha levels, which were increased in BHD (from 10.9 +/- 3.1 to 14.7 +/- 4.1 pg/mL; p=0.004) and reduced in PHF-AF (from 11.9 +/-2.8 to 6.3 +/- 2.2 pg/mL; p=0.0004). CONCLUSION: Although the cytokine levels were unchanged during the study in both BHD and PHF AF, the modification of TNF-alpha during the dialysis session was considered as inflammatory significance. PMID- 15747296 TI - [Exogenous vs endogenous reinfusion in HDF]. PMID- 15747297 TI - [Hemodiafiltration with endogenous reinfusion]. PMID- 15747299 TI - [Standard hemodiafiltration]. PMID- 15747298 TI - [Chronic inflammation]. PMID- 15747300 TI - [MIA syndrome and nutritional status]. PMID- 15747301 TI - [The purity of dialysis fluids: Guidelines of Piedmont and Aosta Valley]. PMID- 15747302 TI - [The quality of dialysis water]. PMID- 15747303 TI - [Innovations in dialytic therapy and daily clinical problems: anemia]. PMID- 15747304 TI - [Metabolic acidosis in hemodialyzed patient: clinical considerations]. PMID- 15747305 TI - [The use of dialysate conductivity in the sodium and urea kinetic models]. PMID- 15747306 TI - [Oxygen saturation and hemocompatibility]. PMID- 15747307 TI - [Contact phase activation can occur with certain types of activated carbon]. AB - HFR is an integrated hemodiafiltration system that utilizes a double chamber filter to separate convection from diffusion. The ultrafiltrate is regenerated by passage through a sorbent cartridge made up of resin and activated carbon. A small percentage of patients using this technique had gastrointestinal symptoms that included nausea/vomiting, diarrhea and/or stomach cramps approximately 1-2 hours after the start of HFR. We undertook a series of investigations to try and elucidate the cause of these reactions. Since the majority of the patients were taking ACE inhibitors, attention was focused on contact phase activation. Healthy and uremic plasma were incubated with different components of the HFR circuit. The activated carbon caused a moderate activation of factor XII and production of kallikrein, while there was no activation for the lines, double filter or resin. Patients taking ACE inhibitors may be at risk for treatments involved with contact phase activation as ACE inhibitors also block the degradation of bradykinin. A new sorbent cartridge has now been developed that contains only resin. PMID- 15747308 TI - [Optimization of a HFR sorbent cartridge for high molecular weight uremic toxins]. AB - HFR is a hemodiafiltration method with regeneration of the ultrafiltrate. It consists of a double chamber filter that separates convection from diffusion. The ultrafiltrate exits from the convective filter, passes through a sorbent cartridge where uremic toxins bind to the sorbent. The "purified" ultrafiltrate is then returned to the patient. This study undertook a series of in vitro and ex vivo experiments to optimize the conditions for maximal adsorption and treatment efficacy. An emphasis was placed on a resin only cartridge as previous studies suggested that some patients may be sensitive to the activated carbon, particularly if they are taking ACE inhibitors. PMID- 15747309 TI - [On-line hemodiafiltration]. PMID- 15747310 TI - [Clinical experience in HFR with a new resin-only sorbent cartridge]. AB - Adsorbent therapies have become increasingly popular over the last several years as they permit an additional method to selectively or non-selectively remove toxins. Adsorbents offer a unique removal strategy as they have an extremely high adsorption capacity due to their great surface area. This paper describes experiments that utilized a synthetic divinylbenzene styrenic resin cartridge to remove uremic toxins from chronic renal failure patients. The resin-only cartridge was tested as an alternative after a small number of patients (primarily taking ACE inhibitors) experienced gastrointestinal problems using hemodiafiltration with on-line regeneration (HFR). Subsequent laboratory evidence suggested that the particular carbon used in the cartridge was able to activate contact phase activation. This could potentially cause problems in patients taking ACE inhibitors, as they are unable to degrade bradykinin efficiently. The resin-only cartridge was tested in at 6 centers throughout Italy and included patients that had experienced previous reactions to the carbon-resin cartridge. At the conclusion of the study, no adverse reactions were reported and the cartridge exhibited excellent removal of b2 microglobulin and angiogenin. PMID- 15747311 TI - [Hemodiafiltration with endogenous reinfusion (HFR): biochemical and gas analytical analysis of the "regenerated" ultrafiltrate]. AB - PURPOSE: During convective techniques, a replacement fluid (R) is necessary that is sterile and pyrogen-free. Using an integrated absorption cartridge, the ultrafiltrate (UF) can be "regenerated"; and used as R. This method is hemodiafiltration with reinfusion (HFR). This study aimed to evaluate the real UF composition after "regeneration" by the resin-charcoal integrated absorption cartridge. METHODS: In eight uremic patients treated with HFR the UF was evaluated at 5, 15, 30, 60, 120, 180 and 240 min after HFR start at the inlet and the outlet resin-charcoal cartridge using the following parameters: urea, creatinine (Cr), uric acid, phosphates, glucose, Beta 2-microglobulin (beta2-m), Na+, K+, Ca++, pH, pCO2, and HCO3-. RESULTS: Blood (%): urea -61.2 +/- 9.7; Cr 55.4 +/- 8.1; uric acid -69.8 +/- 9.3; phosphates -31.8 +/- 15.7; glucose -8.4 +/ 20.5; Beta2-m -60.3 +/- 11.1; pH +0.76 +/- 0.58; pCO2+ 3.3 +/- 8.5; HCO3- +18.1 +/- 13.5. In UF (outlet vs inlet): urea was not adsorbed; Cr and uric acid were adsorbed; phosphates were not adsorbed; glucose was partially adsorbed (only in the 1st 90 min); Beta2-m was almost totally adsorbed; Na+ and K+ were not adsorbed; for pH, pCO2, and HCO3- there were no significant variations between the inlet and the outlet. CONCLUSIONS: HFR seems to be an easy-to-perform hemodiafiltration (HDF) technique, capable of resolving the typical problems of availability and the production of sterile and ultrapure reinfusion solution. PMID- 15747312 TI - [On-line haemodiafiltration with sorbent-regenerated ultrafiltrate as replacement fluid: Beta2-microglobulin removal versus filtration fraction]. AB - PURPOSE: In order to encourage the removal of middle molecules in hemodiafiltration (HDF) techniques an attempt is made to maximize convective clearance by increasing the ultrafiltration rate. However, convective clearance is limited by the maximum filtration fraction (FF%) obtainable, by the pre- or post reinfusion method and by the convective surface and the capacity of the filter used. This study aimed to evaluate the effect of the FF% in the removal of Beta2-microglobulin (Beta2-m) during hemodiafiltration reinfusion (HFR) to identify the best ultrafiltration strategies; and therefore, a better removal of medium molecular weight solutes in this hemodiafiltrative technique recently introduced in clinical practice. METHODS: Ten chronic uremic patients (eight males, two females; age 66 +/- 18 yrs) already on renal dialysis therapy (RDT) for 80 +/- 36 months, were subjected to four HFR sessions, with Td=240 +/- 10 min, Qb=312 +/- 18 and Qd=500 mL/min; the reinfusion rates (Qr) used were 43.6 +/ 7.2 mL/min (25-58) with FF% rates varying from 20-34 (24.2 +/- 3.8) for hematocritic levels of 34.6 +/- 4.2% at the start of the dialysis session. For each session the intradialytic reduction rates (RR%) of urea, creatinine (Cr), phosphate, uric acid and Beta2-m and its average clearance (KBeta2-m mL/min) were evaluated. RESULTS: The results obtained gave a RR% for urea of 69.4 +/- 5 (Kt/Veq=1.23 +/- 0.2) and for Cr, phosphate and uric acid values of, respectively, 61.2 +/- 5.4, 47.5 +/- 10 and 75.8 +/- 6.7. The intradialytic reductions in Beta2-m were 49.3 +/- 10.3% with a variability range from 29-69% and with average KBeta2-m values of 63.8 +/- 13.5 mL/min. The RR% of ss2-m and KBeta2-m were inversely correlated (p<0.01) to the FF% rate applied during the treatment; 75% of the HRF sessions in which we observed a reduction in Beta2-m levels >40% were those where a FF% between 20 and 26% was applied. CONCLUSIONS. From our study, it appears that in HFR the best ultrafiltration strategy from the convective sector in removing Beta2-m has FF% values in the range 20-26%. The occurrence of lower intradialytic reductions of Beta2-m with increasing FF% can be interpreted as a consequence of phenomena related to high intradialytic hemoconcentrations, to the excessive increase in the TMP and/or the increase in the protein cake with a consequent reduction in permeability and mass transfer. Although using a limited convective surface with a limited possibility of increasing the FF%, nevertheless, HFR seems capable of ensuring a satisfactory uremic toxin removal of low and medium molecular weight, which combined with the high biocompatibility deriving from the use of reinfused endogen, can be considered an effective dialytic strategy for preventing or retarding the complications in dialytic patients. PMID- 15747313 TI - [HFR vs HDF-ON line: plasmatic amino acids loss evaluation]. AB - PURPOSE: The high convection dialytic techniques, such as hemodiafiltration (HDF), can cause the loss of important molecules such as growth factors, vitamins and amino acids. Hemodiafiltration reinfusion (HFR) is an HDF on-line process, using a sipping cartridge, able to remove uremic toxins and give back a "repaired" ultra-filtrate to the patient. We aimed to establish the plasmatic amino acid levels before and after dialysis in HFR vs. HDF on-line, with scrupulous attention to branched chain amino acids (BCAA) such as isoleucine, leucine and valine. These amino acids, often present with low plasmatic levels in hemodialyzed patients, seem to be related to a picture of malnourishment. METHODS: Eleven male patients on bicarbonate dialysis, for at least 1 yr, were evaluated (average dialytic age = 88 months, /average age = 67 yrs), with good dialytic efficiency and body mass levels, randomized in HFR or HDF on-line (filter PAN AN 69) for 1 week of treatment, respectively. The different results of each method were controlled for the same patient. Blood samples were taken before and after dialysis in each 2nd hemodialytic weekly session. Total amino acids, essential, non-essential and BCAA were determined by gas-chromatography. RESULTS: There was no difference detected in pre-dialytic plasmatic levels of analyzed amino acids between the two groups. In post-dialysis, HDF patients demonstrated a total essential, non-essential amino acid and BCAA higher loss rate, compared to HFR patients. Post-dialysis amino acid level averages were: total amino acids in HDF 1852 +/- 302.6 micromol/L, in HFR 2395 +/- 492.8 micromol/L (p = 0.018); essential amino acids in HDF 428.8 +/- 118.2 micromol/L, in HFR 510.3 +/- 129.3 micromol/L (p = 0.022); non-essential amino acids in HDF 1176 +/- 213 micromol/L, in HFR 1546 +/- 339.2 micromol/L (p = 0.01); BCAA in HDF 242.7 +/- 83.42 micromol/L, and in HFR 286.7 +/- 89.9 micromol/L (p = 0.03). CONCLUSIONS: Since low plasmatic BCAA levels are related to anorexia and malnourishment, the loss of these amino acids can be important in the dialytic technique choice. HFR can offer an outstanding advantage, combining a high convection treatment with medium molecule removal, without compromising physiologic molecule loss. PMID- 15747314 TI - [Biocompatibility of the acetate in the dialysis fluid]. AB - PURPOSE: Dialytic vasculopathy is a major morbidity and mortality risk factor in patients undergoing chronic dialysis treatment. Among the pathogenetic factors some are related to the uremic condition, others are due to biocompatible reactions to dialytic materials. Endothelial cells (EC) are the target of the mediators released during bioincompatible reactions, and the related effects could be considered the initial event eliciting the vasculopathy pathogenesis. Among the others, we focused our attention on the role played in this process by the inducible isoform of nitric oxide (NO) synthase (iNOS). In previous studies we demonstrated that bioincompatible membranes, as well as acetate-containing dialysis buffers stimulate iNOS gene expression and activity in endothelial cells in culture. In this study, we planned to evaluate the potential role of a new dialysis buffer in which acetate has been substituted with HCl as a stabilizer. METHODS: ECs were incubated for 12 h at 37 degrees C with different dialysis buffers: acetate (Acet), standard bicarbonate (Bic), acetate-free buffer (AF) and HCl-bicarbonate (BicHCl). We evaluated in reverse transcriptase polymerase chain reaction (RT-PCR) the gene transcription for iNOS, the NOS activity (as the production of H3 citrulline from H3 arginine by ionic exchange chromatography), EC proliferative (H3 thymidine incorporation) and pro-apoptotic rate (TUNEL analysis) and the nuclear translocation of the transcriptional factor NF-kappaB (EMSA). RESULTS: Acetate, even in the low concentration present in Bic was able to induce a significant iNOS gene transcription (results expressed as relative units and referred to basal values: Acet 1.9 +/- 0.01 fold increase, p<0.01; Bic 1.45 +/- 0.03 p<0.05; BicHCl 1.24 +/- 0.01; AF 1.17 +/- 0.02) and translation. Acetate at concentrations both of 3 mmoL and 38 mmoL (present in the bicarbonate buffer) significantly increased the enzymatic NOS activity vs unconditioned ECs: Acet 3.46 +/- 0.3, p<0.0005; Bic 1.69 +/- 0.2, p<0.005; BicHCl 1.24 +/- 0.15; AF 1.17 +/- 0.05. The EC proliferative index was significantly depressed by acetate containing dialysis buffers (unconditioned ECs 100%, Acet 38 +/- 15%, p<0.01; Bic 65 +/- 6%, p<0.05; AF 87 +/- 8%; BicHCl 75 +/- 6%). The percentage of apoptotic ECs was significantly increased by buffers contain-ing Acet vs BicHCl and AF. Finally, acetate at the concentrations present in Acet and Bic activated and promoted the nuclear translocation of the transcriptional factor NF-kappaB in ECs (p<0.01 vs unconditioned cells). CONCLUSIONS: The acetate-free dialysis buffers have better biocompatibility and potentially down-modulate the flogistic and sclerotic processes responsible for dialytic vasculopathy. PMID- 15747316 TI - A robust clustering approach for NMR spectra of natural product extracts. AB - A robust method was developed to cluster similar NMR spectra from partially purified extracts obtained from a range of marine sponges and a plant biota. The NMR data were acquired using microtiter plate NMR (VAST) in protonated solvents. A sample data set which contained several clusters was used to optimize the protocol. The evaluation of the robustness was performed using three different clustering methods: tree clustering analysis, K-means clustering and multidimensional scaling. These methods were compared for consistency using the sample data set and the optimized methodology was applied to clustering of a set of spectra from partially purified biota extracts. PMID- 15747315 TI - [On-line hemodiafiltration without acetate]. AB - PURPOSE: The suitably filtered dialysate which is currently reinfused during on line hemodiafiltration (HDF-OL) contains bicarbonate and small doses of acetate. The trend of acetataemia During "forced" convective treatments has never been studied. The gain in acetate secondary to the considerable quantities of fluids infused might have clinical significance in relation to the well-known side effects of this anion. METHODS: In this pilot study 12 patients underwent HDF-OL with reinfusion in predilution of 40 L of substitution fluids containing or not 3 mmol/L of acetate. Apart from this variable, all the other treatment parameters were the same in both procedures. The treatments were carried out in two short consecutive intervals in a random sequence. RESULTS: During HDF-OL the use of dialysate containing small doses of acetate is associated with levels of acetataemia 5-6 times higher compared to the basal. HDF-OL without acetate cancels out this increase. The acetate gained by the patients is significant, on average 75 mmol, and accounts for over 1/3 of the global base gain. Consequently, the bicarbonataemia levels at the end of treatment are significantly higher in HDF-OL with acetate than in the treatment without. Two hours after the end of the treatments the IL-6 levels tend to grow in both methods, but numerically less in HDF-OL without acetate; the difference verges on meaningfulness. CONCLUSIONS: The acetate gain is significant during forced convective treatments carried out with standard dialysate. This acetate gain can trigger cytokinin activation. These events are cancelled out by eliminating the acetate from the dialysate. The absence of this anion will be compensated with appropriate increases in the concentration of bicarbonate in the dialysis fluid. PMID- 15747317 TI - Determination of absolute configurations of chiral molecules using ab initio time dependent Density Functional Theory calculations of optical rotation: how reliable are absolute configurations obtained for molecules with small rotations? AB - The absolute configuration (AC) of a chiral molecule can be determined via calculation of its specific rotation. Currently, the latter is most accurately carried out using the TDDFT/GIAO methodology. Here we examine the reliability of this methodology in determining ACs of molecules with small specific rotations. We report TDDFT/GIAO B3LYP/aug-cc-pVDZ//B3LYP/6-31G* calculations of the sodium D line specific rotations, [alpha]D, of 65 conformationally rigid chiral molecules whose experimental [alpha]D values are small (<100). The RMS deviations, sigma, of calculated and experimental [alpha]D values is 28.9. The distribution of deviations is approximately Gaussian, i.e., random. For eight molecules, more than 10% of the set, the sign of the predicted [alpha]D is incorrect. In determining an AC of a rigid molecule from [alpha]D with 95% confidence, the calculated [alpha]D value must lie within +/-2sigma of the experimental [alpha]D for one, but not both, of the possible ACs. For the 65 molecules of this study +/ 2sigma is 57.8. For conformationally flexible molecules, the error bar is +/- >57.8. PMID- 15747318 TI - The influence of 1-aminocyclopentane-1-carboxylic acid at position 2 or 3 of AVP and its analogues on their pharmacological properties. AB - This study describes the synthesis and some pharmacological properties of eight new analogues of arginine vasopressin (AVP) substituted at position 2 or 3 with cycloleucine (1-aminocyclopentane-1-carboxylic acid, Apc). All new peptides were tested for their pressor, antidiuretic and uterotonic in vitro potency. The Apc3 modification resulted in an almost complete loss of potency in all three tests, which is interpreted as a loss of interaction with all three neurohypophyseal hormone receptors. On the other hand, the Apc2 modification resulted in compounds having differently modified activities (high antidiuretic potency, low and graded pressor activity and either no activity or low oxytocin antagonizing activity in the uterotonic in vitro test) thus selectively altering the interaction with the receptors similar to that of 1-aminocyclohexane-1-carboxylic acid (Acc). The results obtained may be helpful for designing new analogues of arginine vasopressin. PMID- 15747319 TI - Studies on intramolecular hydrogen bonding between the pyridine nitrogen and the amide hydrogen of the peptide: synthesis and conformational analysis of tripeptides containing novel amino acids with a pyridine ring. AB - For the first time tripeptides, Z-AA(1)-Xaa-AA(3)-OMe (AA(1) and AA(3) = Gly or Aib, Xaa = 2Pmg and 2Pyg) were prepared containing alpha-methyl-alpha-(2 pyridyl)glycine (2Pmg) and alpha-(2-pyridyl)glycine (2Pyg) by solid-phase Ugi reaction. These results clearly indicate that for the preparation of tripeptides containing an amino acid with a pyridine ring, the solid-phase Ugi reaction is very useful.NMR analysis clarified that 2Pmg-containing tripeptides adopt a unique conformation with an intramolecular hydrogen bond between 2Pmg-NH and the pyridine nitrogen. However, in the case of Z-Gly-2Pyg-Gly-OMe, the intramolecular hydrogen bonding between 2Pyg-NH and the pyridine nitrogen was not observed, whereas Z-Aib-2Pyg-Aib-OMe adopts a unique conformation with an intramolecular hydrogen bond between 2Pyg-NH and a pyridine nitrogen. Conformational analysis of the tripeptides, Z-AA(1)-Xaa-AA(3)-OMe (AA(1), AA(3) = Gly or Aib, Xaa = alpha,alpha-di(2-pyridyl)glycine (2Dpy), alpha-phenyl-alpha-(2-pyridyl)glycine (2Ppg), 2Pmg and 2Pyg), clarified that when an alpha,alpha-disubstituted glycine with a 2-pyridyl group at an alpha-carbon atom is introduced into any peptide, an intramolecular hydrogen bond between a pyridine nitrogen and an amide proton is formed and conformational mobility of the peptide backbone is restricted. PMID- 15747320 TI - Absolute stereochemistry of unusual biopolymers from Ascomycete culture LL-W1278: examples that derivatives of (S)-6-hydroxymellein are also natural fungal metabolites. AB - The new linear polyesters, W1278-A, B, and C, were isolated from culture extracts of a freshwater Ascomycete fungus. Their structures were elucidated on the basis of spectroscopic analyses and chemical transformations, with the absolute configuration established by the circular dichroism (CD) method using (R)-(-)-6 hydroxymellein as reference. Unexpectedly, W1278-A, B, and C contain (S)-6 hydroxymellein residues as concluded from the display of the opposite ellipticity than that of known (3R)-(-)-3,4-dihydro-6,8-dihydroxy-3-methylisocoumarin as revealed by CD. PMID- 15747321 TI - Certification of the cadmium content in certified reference materials for Cd rice flour. AB - The materials used to prepare the certified reference materials (CRMs) for Cd rice flour described here were taken from Cd-contaminated rice samples, round grained and non-sticky rice grown in different provinces throughout China. The Cd rice materials were first blown to remove dust and husk, and then were successively ground, sieved, dried and mixed to form homogeneous rice flour. The Cd rice flour was packed in glass bottles, each bottle containing 40 g. All bottled CRMs were subjected to 60Co radiation (2.5 megarads) for long-term preservation. Atomic absorption spectrometry was used to determine the homogeneity of the cadmium content in each material. The certification of the cadmium content for the three CRMs was performed using isotope dilution mass spectrometry. PMID- 15747322 TI - Photodimerization in pyrimidine-substituted dipeptides. AB - Ten N(epsilon)-glycylornithineamide derivatives have been synthesized containing various N(alpha)-linked pyrimidine-1-ylacetyl groups which can undergo (2pi + 2pi) photodimerization on irradiation with UV light at 254 nm. The dimerization efficiency of the free and bound pyrimidine groups was compared in aqueous solution: it was dependent on the substitution of the pyrimidine ring. N(alpha),N(alpha')-bis-(uracil-1-ylacetyl)-(N(epsilon)-glycylornithineamide) and N(alpha),N(alpha')-bis-(5-bromouracil-1-ylacetyl)-(N(epsilon) glycylornithineamide) were identified as possible candidates for optical data storage. PMID- 15747323 TI - Identification of oral antidiabetics and their metabolites in human urine by liquid chromatography/tandem mass spectrometry--a matter for doping control analysis. AB - Since 1999, insulin belongs to the list of prohibited substances of the International Olympic Committee and the World Anti-Doping Agency. Except for patients suffering from insulin-dependent diabetes mellitus, the administration of insulin is not allowed. Therapeutics developed to treat non-insulin-dependent diabetes mellitus act as releasing factors of endogenously produced insulin or improve its efficiency mediating the glucose uptake into insulin-dependent tissues. Hence, these compounds are also relevant for sports drug testing, and a fast, robust, and sensitive assay was developed to identify 12 oral antidiabetic agents or respective hydroxylated metabolites in human urine. Urine specimens are enzymatically hydrolyzed; target analytes are extracted by liquid-liquid extraction and identified by means of liquid chromatography interfaced to tandem mass spectrometry by electrospray ionization. Detection limits of respective drugs ranged between 10 and 30 ng/mL, metabolites of therapeutics were characterized by diagnostic fragmentation pathways upon collisionally activated dissociation of protonated molecules, and general fragmentation routes were proposed. PMID- 15747324 TI - Probabilistic model for estimating field mortality of target and non-target bird populations when simultaneously exposed to avicide bait. AB - A probabilistic model was developed to estimate target and non-target avian mortality associated with the application of the avicide CPTH (3-chloro-p toluidine hydrochloride) to minimize sprouting rice damage in the southern USA. CPTH exposures for individual birds were predicted by random sampling from species-specific non-parametric distributions of bait seed consumption and CPTH residues detected on individual bait seeds. Mortality was predicted from the species-specific exposure versus mortality relationship. Individual variations in this response were captured in the model by Monte Carlo sampling from species specific distributions of slopes and median toxicity values (LD50) for each bird. The model was used to simultaneously predict mortality (percentage of exposed population and number of birds killed/weight of consumed bait) for a target (blackbird) and non-target (mourning dove) species feeding on bait sites for up to five consecutive days. PMID- 15747325 TI - Improved matrix-assisted laser desorption/ionisation sample preparation of a partially depolymerised cellulose derivative by continuous spray deposition and interfacing with size-exclusion chromatography. AB - Continuous spray deposition (CSD) of aqueous solutions of partially depolymerised methyl cellulose was found to improve matrix-assisted laser desorption/ionisation (MALDI) sample preparation. One feature was that the sensitivity in MALDI time-of flight mass spectrometry increased up to an order of magnitude compared with the standard sample preparation method. Another feature was that CSD provided targets for MALDI with homogeneously distributed analyte. This resulted in a more even signal intensity and a higher reproducibility than in the standard method. High mass discrimination was more pronounced in CSD than in the standard method. Size exclusion chromatography with aqueous eluent was coupled online to CSD onto matrix-precoated foils. The suitability for determination of the molar mass distribution of methyl cellulose was investigated. PMID- 15747326 TI - Prenatal screening for congenital heart disease using real-time three-dimensional echocardiography and a novel 'sweep volume' acquisition technique. AB - OBJECTIVES: Conventional prenatal screening for congenital heart disease (CHD) involves a time-consuming and highly operator-dependent acquisition of the four chamber view and outflow tracts. By acquiring the entire fetal heart instantaneously as a single volume, real-time three-dimensional echocardiography (RT3DE) may facilitate fetal cardiac screening. METHODS: Four reviewers, each experienced with fetal cardiac imaging, blindly and independently evaluated a single cardiac volume from each of 18 fetuses (11 normal, seven with CHD). Two dimensional echocardiography served as the gold standard. Three-dimensional evaluation of each fetus included a series of volume acquisitions lasting 2-6 s each. A 'sweep volume' technique was developed to fit larger hearts into a single non-gated volume. RESULTS: RT3DE had a high sensitivity for detecting CHD (93%), with only a single case being missed by two observers. Specificity for CHD was low (45%), with a high rate of 'cannot determine' responses and false positive artifacts. CONCLUSIONS: These preliminary results suggest that RT3DE has the potential to function as a screening tool for fetal heart disease. However, artifacts must be recognized and minimized, resolution must improve, and substantial training will be necessary prior to widespread clinical use. PMID- 15747327 TI - Cardiac diverticulum with pericardial effusion: report of two new cases treated by in-utero pericardiocentesis and a review of the literature. AB - Congenital cardiac diverticula are rare abnormalities that may occur as isolated malformations. They are often associated with pericardial effusions, which may cause both pulmonary hypoplasia and progressive fetal hydrops. Few cases are reported in fetal life. Mount Sinai Hospital, Toronto, has previously reported two cases of cardiac diverticula complicated with pericardial effusion successfully treated in utero with aspiration of the pericardial fluid. Here a further two cases of isolated apical right ventricular diverticula with large pericardial effusion, one diagnosed at 16 weeks and another at 13 weeks' gestation are described. In-utero drainage of pericardial effusion was performed once in each case at 16 and 14 weeks' gestation, respectively, with good neonatal outcome. Both had normal karyotype and there was no evidence of maternal or fetal infection. The pericardial effusion did not recur in either case. Given the otherwise favorable prognosis for this lesion, and the excellent response in these cases, prenatal pericardiocentesis should be considered in similar cases. PMID- 15747328 TI - Structural assignment of isomeric 2-aminopyridine-derivatized oligosaccharides using negative-ion MSn spectral matching. AB - To investigate the possibility of structural assignment based on negative-ion MS2 spectral matching, three isomeric pairs of 2-aminopyridine (PA)-derivatized non fucosylated, fucosylated, and sialylated oligosaccharides (complex type N glycans) were analyzed using high-performance liquid chromatography/ion trap mass spectrometry (HPLC/ITMS) with a sonic-spray ionization (SSI) source. In the SSI negative-ion mode the deprotonated molecule [M-2H]2- becomes prominent. Negative ion MS2 spectra derived from such ions contain many fragment types (B and Y, C and Z, A, and D) and therefore are more informative than the positive-ion MS2 spectra derived from [M+H+Na]2+ ions, which usually consist mainly of B and Y fragment ions. In particular the internal ions (D- and E-type ions) provided useful information about the alpha1-6 branching patterns and the bisecting GlcNAc residue. Spectral matching based on the correlation coefficients between negative ion MS2 spectra was performed in a manner similar to the positive-ion MS2 spectral matching previously reported. It was demonstrated that negative-ion MS2 spectral matching is as useful and applicable to the structural assignment of relatively large non-fucosylated, fucosylated, and sialylated PA-oligosaccharide isomers as its positive-ion counterpart. PMID- 15747329 TI - Mass spectrometric study of substituted alpha-acylaminocinnamic acids derived from partial hydrolysis of 5(4H)-oxazolones. PMID- 15747330 TI - Fetal congenital cardiac diverticulum with pericardial effusion: two cases with different presentations in the first trimester of pregnancy. AB - Two cases of fetal congenital cardiac diverticula diagnosed at 14 and 12 weeks of gestation are reported. Both presented with a large pericardial effusion. One fetus presented with increased NT which evolved into fetal hydrops. Pericardiocentesis at 16 weeks led to progressive resolution of hydrops. The other showed no hemodynamic compromise, normal nuchal translucency (NT) and spontaneous regression of the effusion. Both children were well at 22 and 17 months of age at the time of writing. PMID- 15747331 TI - Effect of post-excitation radius on ion abundance, mass measurement accuracy, and isotopic distributions in Fourier transform ion cyclotron resonance mass spectrometry. AB - We report an evaluation of a modern Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) instrument to determine the general trend of post excitation radius on total ion abundance, mass measurement accuracy, and isotopic distributions for internally calibrated mass spectra. The optimum post-excitation radius was determined using total ion abundance, mass measurement accuracy (MMA), and isotope ratios. However, despite the utility of internal calibration for achieving ultimate MMA, the internal calibrant ions were insufficient for compensating for sub-optimum ICR cell conditions. The findings presented herein underscore the importance of determining the optimal post-excitation radius in FT ICR-MS to achieve high ion abundance (low limits of detection), high MMA, and valid isotopic distributions. PMID- 15747332 TI - Gas chromatography/mass spectrometry determination of amphetamine-related drugs and ephedrines in plasma, urine and hair samples after derivatization with 2,2,2 trichloroethyl chloroformate. AB - A new analytical approach, based on derivatization with 2,2,2-trichloroethyl chloroformate and gas chromatography/mass spectrometry (GC/MS), was investigated for qualitative and quantitative analyses of a large range of amphetamine-related drugs and ephedrines in plasma, urine and hair samples. Sample preparation involved alkaline extraction of analytes from biological samples using Extrelut columns, after addition of the internal standard 3,4 methylenedioxypropylamphetamine (MDPA), and subsequent derivatization to produce 2,2,2-trichloroethylcarbamates. GC/MS analyses, in splitless mode using a slightly polar 30-m capillary column, were performed with quadrupole or ion trap instruments. MS acquisition modes were electron ionization (EI) in full-scan or selected ion monitoring (SIM) modes (quadrupole), and full-scan MS or MS/MS modes with chemical ionization (CI) conditions (ion trap). EI spectra of 2,2,2 trichloroethylcarbamates showed variably abundant molecular ions as well as abundant diagnostic fragment ions, both characterized by ion clusters reflecting the isotope distribution of three chlorine atoms in the derivatized molecules. CI spectra showed abundant protonated molecules. Quantitative studies using EI SIM conditions gave recoveries in the range 74-89%, linear response over ranges of 10 2000 ng/mL (plasma and urine) and 0.20-20 ng/mg (hair), with corresponding limits of detection in the ranges 2-5 ng/mL and 0.1-0.2 ng/mg. Potential applications (following full method validation) include clinical and forensic toxicology, as well as doping control. PMID- 15747333 TI - Pulmonary function impairment in children following hematopoietic stem cell transplantation. AB - BACKGROUND: Deterioration of pulmonary function after hematopoietic stem cell transplantation (SCT) is a well-known late effect of this treatment, but the course of pulmonary function over time is less clear. The aim of our study was to establish both the prevalence and course of pulmonary function abnormalities in children following SCT. METHODS: Thirty-nine of 106 patients, who visited a post SCT late effects clinic and who underwent a pulmonary function test (PFT) both before and at least twice after SCT were included in this study. Forced expiratory volume in 1 sec (FEV1), forced vital capacity (FVC), total lung capacity (TLC), and total lung diffusion capacity (TLCO) were determined and recorded as percentage predicted for age, sex, and length matched controls. Values of less than 80% of predicted were considered abnormal. Change in PFT parameters over time was determined by comparing the mean PFT parameter in our group at three different time points: pre-SCT, < or =1 year post-SCT (SCTpost1) and >1 year post-SCT (SCTpost2). RESULTS: After SCT restrictive and/or diffusion abnormalities are most prevalent (45% and 76% at SCTpost1, respectively). A significant decrease of TLC (-9.7%) and TLCO (-20.3%) was observed during the first year after SCT, with improvement over time, but no normalization. Obstructive lung disease was less common (6% at SCTpost1). Clinical signs of lung function impairment were rare. CONCLUSIONS: Restrictive and diffusion lung function disorders are common after SCT. They improve over time but do not normalize. As only a few patients with pulmonary function abnormalities had clinical signs of lung function impairment, the clinical relevance of performing long-term follow-up of PFT is questionable. PMID- 15747334 TI - Low risk episodes of fever and neutropenia in pediatric oncology: Is outpatient oral antibiotic therapy the new gold standard of care? PMID- 15747335 TI - Role of exercise in rehabilitation of cancer survivors. AB - Approximately 70% of all children diagnosed with malignancy become long-term survivors. Anthracycline cardiotoxicity may complicate physical performance and invite a "spectrum of disuse." There is a paucity of information derived from longitudinal, prospective, randomized clinical trials of exercise in cancer survivors, and there are no guidelines concerning the risk of recreational (non competetive) exercise in adolescent and young adult patients with anthracycline cardiomyopathy. This review will discuss screening procedures that should be performed when assigning exercise regimens to cancer survivors with or without evidence of left ventricular dysfunction. Current recommendations for exercise testing and exercise prescription are also provided. PMID- 15747336 TI - Menopausal hormone therapy: currently no evidence for cardiac protection. AB - Coronary heart disease is the leading cause of death among women of older age. Interest emerged as to whether premenopausal endogenous estrogen conferred cardiac protection and whether menopausal hormone therapy might prove beneficial. The results of pivotal clinical trials including the randomized, placebo controlled hormone trial of the Women's Health Initiative (WHI) and the Heart and Estrogen/progestin Replacement Study (HERS) have led to new Guidelines from the American Heart Association (AHA) that state that hormone therapy should not be initiated or continued for the primary or secondary prevention of cardiovascular disease in menopausal women. PMID- 15747337 TI - Sickle cell hepatopathy: clinical presentation, treatment, and outcome in pediatric and adult patients. AB - BACKGROUND: Standard diagnostic criteria and therapy are lacking for sickle cell hepatopathy, an uncommon complication of sickle cell disease. Here we propose diagnostic and therapeutic guidelines based on our experience and on reported cases. METHODS: We defined sickle hepatopathy by a total serum bilirubin concentration >13 mg/dl not explained by severe acute hemolysis, viral hepatitis, extrahepatic obstruction, or hepatic sequestration. We reviewed the records of all children with sickle hepatopathy seen at our institution during the past 20 years and the reported cases from the literature. Patients were categorized into two groups based on whether hepatic dysfunction at presentation was mild (Group I) or severe (Group II). RESULTS: Seven patients were identified from our institution and 37 patients from the literature. The 44 patients were evenly divided between the two groups. Group I patients had a significantly lower mean age (11.8 years vs. 24.5 years, P = 0.0001), maximum bilirubin level (36.2 mg/dl vs. 76.8 mg/dl, P = 0.0008), and frequency of treatment with exchange transfusions (P = 0.03). Overall, mortality was 4% in Group I and 64% in Group II (P = 0.0001). Gender and recurrence rate did not differ. Among Group II patients, only two of nine who received exchange transfusion died, whereas 12 of 13 who did not receive exchange transfusion died (P = 0.0015). CONCLUSIONS: Sickle cell hepatopathy is an uncommon complication characterized by extreme hyperbilirubinemia and either mild or severe hepatic dysfunction. Children and adults can present with either form; however, adults have a higher frequency of the severe form. Exchange transfusion may be the only effective management for initial episodes of severe sickle cell hepatopathy. PMID- 15747338 TI - Biology, clinical characteristics, and management of adrenocortical tumors in children. AB - Childhood adrenocortical tumors (ACT) are very aggressive endocrine neoplasms whose incidence is quite low. Little is known about their pathogenesis, clinical presentation, and optimal treatment. In recent years, however, new information has been derived from the International Pediatric Adrenocortical Tumor Registry (IPACTR), and new clues to its pathogenesis have emerged. To provide an overview of the available data that may apply to pediatric ACT, we reviewed the epidemiology, pathogenesis, and treatment of ACT in adults and in children. Germline TP53 mutation is almost always the predisposing factor in childhood ACT. A unique germline mutation (TP53-R337H) has been described in Southern Brazil, where the incidence of ACT is 10-15 times the general incidence. Childhood ACT typically present during the first 5 years of life and has female predominance. Hormone hyperproduction is almost universal, and most patients present with virilization. Two-thirds of patients have resectable tumors. Surgery is the definitive treatment for ACT, and a curative complete resection should always be attempted. Cisplatin-based chemotherapy with mitotane is indicated for unresectable or metastatic disease, although its impact on overall outcome is slight. In childhood ACT, age, tumor size, and tumor resectability are the most important prognostic indicators. Outcome is stage-dependent; patients with small, resectable tumors have survival rates in excess of 80%, whereas the outcome for patients with unresectable disease is dismal. Patients with large, resectable tumors have an intermediate outcome. Childhood ACT are rare, but their unique epidemiology appear to implicate novel oncogenic pathways that are unique to the pediatric population. Multi-institutional and prospective studies are necessary to further our understanding of the pathogenesis and to improve outcomes. PMID- 15747339 TI - Re: Slack M. Culligan P, Tracey M, Hunsicker K, Patel B, Sumeray M. 2004. Relationship of urethral retro-resistance pressure to urodynamic measurements and incontinence severity. PMID- 15747340 TI - Test-retest reliability of four questionnaires for patients with overactive bladder: the overactive bladder questionnaire (OAB-q), patient perception of bladder condition (PPBC), urgency questionnaire (UQ), and the primary OAB symptom questionnaire (POSQ). AB - AIMS: This study examined test-retest reliability of four patient-reported outcome measures for patients with overactive bladder (OAB): Overactive Bladder Questionnaire (OAB-q), Patient Perception of Bladder Condition (PPBC), Urgency Questionnaire (UQ), and Primary OAB Symptom Questionnaire (POSQ). METHODS: Patients recruited from urology clinics were scheduled for two visits 2 weeks apart and completed all questionnaires at both visits. A demographic form was completed at Visit 1; and a treatment effect scale was completed at Visit 2. Test retest reliability was examined among stable patients using intraclass correlations (ICC), Spearman's correlations, paired t-tests, Feldt's statistic, and kappas. RESULTS: A total of 47 patients enrolled (mean age = 66.0 years, 74.5% female), with 46 completing both visits; 35 were classified stable. Statistically significant correlations were present between Visits 1 and 2 (P < 0.05) for all subscales of the OAB-q, UQ, and POSQ. Subscale ICCs were moderate to high (OAB-q > or = 0.83, UQ > or = 0.46, POSQ continuous items > or = 0.68). No significant differences between Visit 1 and 2 were noted, except for the OAB-q symptom bother scale (change of 5.8 points on a 100-point scale). The multi-item subscales of the OAB-q and the UQ demonstrated good internal consistency (Cronbach's alpha > or = 0.83 for all subscales) across both visits. Test-retest reliability of the PPBC was somewhat weaker than the other three measures, but still acceptable for use as a global, single-item outcome measure. CONCLUSIONS: The OAB-q, POSQ, and UQ demonstrated good test-retest reliability, with ICCs roughly equivalent or superior to those previously reported for 7-day micturition diaries. Findings suggest that the four measures examined in this study demonstrate the necessary reproducibility for use as outcome measures for OAB treatments. PMID- 15747342 TI - Application of the Blaivas-Groutz bladder outlet obstruction nomogram in women with urinary incontinence. AB - AIMS: To test the applicability of the Blaivas-Groutz nomogram for female bladder outlet obstruction in urinary incontinent women presenting in a general gynecologic practice and to determine how the nomogram results related to the presence of obstructive symptoms according to a standardized questionnaire. METHODS: All women with complaints of urinary incontinence underwent multichannel urodynamic testing, free uroflowmetry and were asked to complete a standardized quality of life questionnaire consisting of the urogenital distress inventory (UDI). The patients were classified according to the Blaivas-Groutz nomogram. RESULTS: One hundred and nine patients were classified according to the nomogram. Thirty-three patients (30.3%) were classified as unobstructed, 63 patients (57.8%) as mildly, 12 patients (11%) as moderately, and 1 patient (0.9%) was classified as severely obstructed. Fifty patients correctly completed the UDI. There was no significant difference (P = 0.61) in the score on the domain UDI obstruction between, according to the nomogram, obstructed and unobstructed patients. Only 18% of the obstructed patients had isolated voiding symptoms suggestive of obstruction. Forty-nine percent of the obstructed patients had urgency-frequency symptoms as well as voiding symptoms suggestive of obstruction. There was no correlation (Pearson, r = -0.06, P = 0.61) between the severity of the symptoms (assessed by the UDI scale) and the degree of obstruction (the four nomogram zones). CONCLUSIONS: Application of the Blaivas-Groutz nomogram gave an unlikely high prevalence of obstruction in our patient group, which showed no correlation with symptoms when measured with the UDI. PMID- 15747341 TI - Pilot study of the feasibility of in-office bladder distention using electromotive drug adminstration (EMDA). AB - PURPOSE: Cystoscopic bladder distention is an important tool in the diagnosis and treatment of interstitial cystitis (IC). We investigated the feasibility of performing bladder distention in the office using two different anesthetic strategies: simple instillation of alkalized lidocaine and electromotive drug administration (EMDA) of lidocaine. MATERIALS AND METHODS: Patients presenting with urinary frequency, urgency, and bladder pain were recruited for an office evaluation protocol which included bladder distention under local anesthesia. An initial group of 10 patients underwent bladder distention after instillation of 5 mg/kg alkalized lidocaine. A second group of 11 patients had lidocaine EMDA anesthesia prior to distention. RESULTS: In the alkaxlized lidocaine group, 6 of the 10 distentions were aborted after less than 5 minutes at only 40 cm H(2)O. In the EMDA group, 7 of 11 of the distentions were completed using 60 cm H(2)O for 7 minutes. EMDA afforded a more effective distention as manifest by a greater percent increase in distention capacity over cystometric capacity compared to alkalized lidocaine (135% vs. 70%). Despite the lower pressure used in the alkalized lidocaine group, the median distention time was only 3 minutes compared to 7 minutes using EMDA. CONCLUSIONS: These results represent the first study of the efficacy of EMDA as local anesthesia for bladder distention compared to another method of anesthesia. Lidocaine EMDA is superior to alkalized lidocaine in that it allows for a greater distention of the bladder for a longer period of time but does not eliminate the pain of bladder distention. PMID- 15747343 TI - Finite-element model construction for the virtual synthesis of the skulls in vertebrates: case study of Diplodocus. AB - The measurement of strains in real skulls is an inductive method that yields information about the stresses occurring in the a priori existing shape. In contrast, the approach taken here to determine the relationship between skull function and skull shape applies Wolff's law through a deductive technique of structure synthesis. This article describes the application of this method in the exact virtual synthesis of a sauropod skull, e.g., Diplodocus longus Marsh from Wyoming. An unspecific homogeneous solid is first constructed, giving the stresses ample volume to spread between points of force application and constraint. ANSYS 7.0 is used to form 10-noded tetrahedral finite elements with a maximum of 130,000 nodes. The initial conditions are the functional spaces for the eye openings, muscle forces, and the placement of the dental arcade, including assumed bite forces. Enforcing equilibrium of forces, the primary 3D stress flows in each load case are summarized by a physiological superposition, which accumulates the highest value of stress in each finite element. If the stress free parts are eliminated and the summarized stress flows are maintained, a reduced model appears, which is very similar to the real skull. This reduction of shape can be repeated iteratively and leads to a more exact form. Changes in the form of the dental arcade, its position relative to the braincase, the origins of muscles, or the height of the face lead to models that clearly resemble morphological differences between genera. The synthesis of a skull in this way demonstrates the direct correlation between functional loading and the biological structure and shape and can be used to test hypotheses regarding the relationship between structure and function during skull evolution. PMID- 15747344 TI - Botulinum A toxin injections into the detrusor: an effective treatment in idiopathic and neurogenic detrusor overactivity? AB - AIMS: To assess and compare the effect of botulinum A toxin (BTX-A) injections into the detrusor in idiopathic and neurogenic detrusor overactivity resistant to anticholinergic treatment. PATIENTS AND METHODS: In a prospective study, 11 patients with idiopathic and 11 with neurogenic detrusor overactivity resistant to anticholinergic treatment were injected with 300 U of BTX-A (Botox) into the detrusor. Clinical and urodynamic parameters were assessed before and after BTX-A injections. RESULTS: In idiopathic as well as in neurogenic detrusor overactivity, median daytime frequency decreased significantly from 11 to 4 (P = 0.004) and 12 to 5 (P = 0.001), median nocturia from 3 to 1 (P = 0.004) and 3 to 1 (P = 0.001), and median number of used pads from 5 to 0 (P = 0.001) and 5 to 0 (P = 0.002), respectively. There was a significant increase in median maximum cystometric capacity from 220 to 340 ml (P = 0.001) and 190 to 410 ml (to instead of) (P = 0.001), median bladder compliance from 20 to 55 ml/cm H(2)O (P = 0.001) and 23 to 60 ml/cm H(2)O (P = 0.004) and median post void residual from 10 to 140 ml (P = 0.002) and 30 to 240 ml (P = 0.002), respectively. Median maximum detrusor pressure decreased significantly from 45 to 29 cm H(2)O (P = 0.002) and 40 to 24 cm H(2)O (P = 0.002), and median detrusor pressure at maximum flow rate from 30 to 14 ml/sec (P = 0.001) and 38 to 21 ml/sec (P = 0.016), respectively. Due to post void residuals >150 ml following BTX-A injections, de novo clean intermittent self-catheterization was necessary in nine patients (four with idiopathic and five with neurogenic detrusor overactivity) and in one patient (with idiopathic detrusor overactivity) a suprapubic catheter was placed. The effect of BTX-A injections lasted for a median time of 5 months in both idiopathic and neurogenic detrusor overactivity. There was no significant difference in idiopathic compared to neurogenic detrusor overactivity in regard to clinical and urodynamic parameters assessed before and after BTX-A injections. CONCLUSIONS: BTX-A injections into the detrusor have a significant and comparable but temporally limited effect in idiopathic and neurogenic detrusor overactivity resistant to anticholinergic treatment. PMID- 15747345 TI - Finite element analysis of the mouse tibia: estimating endocortical strain during three-point bending in SAMP6 osteoporotic mice. AB - To support future studies of tibial bending in a murine model of senile osteoporosis (SAMP6), we sought to determine the relationship between applied external bending force and peak endocortical strain in the tibiae of SAMP6 and control SAMR1 mice. The lower hindlimbs of mice were loaded by three-point bending in the lateral-medial plane with a support length of 10 mm. Force periosteal strain relations were first determined using standard strain gauge methods. Finite-element analysis (FEA) models of the tibia-fibula were generated based on microcomputed tomography images. After choosing appropriate boundary conditions, FEA predictions of periosteal strains were within 15% of measured values. FEA revealed a narrow (3-4 mm) region of the central tibia with well developed bending strains (tension medially, compression laterally); outside this region, we observed high shear strains. Both the strain gauge data and the finite element simulations indicated that the tibia of the SAMP6 mouse was 20-25% stiffer than the SAMR1 tibia, consistent with a larger moment of inertia and higher cortical bone modulus. Thus, higher levels of force are required to produce the same target values of strain in the SAMP6 tibia. The ratio of periosteal to endocortical strain in the region of interest was similar for the two mouse strains (1.5-1.6). Based on these ratios, we scaled the strain gauge data to estimate the force-endocortical strain relations for the two mouse strains. In conclusion, FEA, with supporting strain gauge measurements, has provided unique insight regarding the strain environment throughout the tibia during three-point bending in mice. PMID- 15747346 TI - Modeling elastic properties in finite-element analysis: how much precision is needed to produce an accurate model? AB - The influence of elastic properties on finite-element analysis was investigated using a finite-element model of a Macaca fascicularis skull. Four finite-element analyses were performed in which the model was assigned different sets of elastic properties. In analysis 1, elastic properties were modeled isotropically using published data obtained from human limb bones. Analyses 2-4 used data obtained from skulls of a closely allied species, M. mulatta, but varied as to how those data were incorporated into the model. In analysis 2, the model was assigned a single set of isotropic elastic properties. In analysis 3, each region within the model was assigned its own set of isotropic elastic properties. Finally, in analysis 4, each region received its own set of orthotropic elastic properties. Although a qualitative assessment indicates that the locations of strain concentrations across the model are broadly similar in all analyses, a quantitative assessment of strain indicates some differences between the analyses. When strain data from the finite-element analyses were compared to strain data derived from in vivo experiments, it was found that the model deformed most realistically using the orthotropic elastic properties employed in analysis 4. Results suggest that finite-element analyses can be adversely affected when elastic properties are modeled imprecisely, and that modelers should attempt to obtain elastic properties data about the species and skeletal elements that are the subjects of their analyses. PMID- 15747347 TI - Comparison of beam theory and finite-element analysis with in vivo bone strain data from the alligator cranium. AB - The mechanical behavior of the vertebrate skull is often modeled using free-body analysis of simple geometric structures and, more recently, finite-element (FE) analysis. In this study, we compare experimentally collected in vivo bone strain orientations and magnitudes from the cranium of the American alligator with those extrapolated from a beam model and extracted from an FE model. The strain magnitudes predicted from beam and FE skull models bear little similarity to relative and absolute strain magnitudes recorded during in vivo biting experiments. However, quantitative differences between principal strain orientations extracted from the FE skull model and recorded during the in vivo experiments were smaller, and both generally matched expectations from the beam model. The differences in strain magnitude between the data sets may be attributable to the level of resolution of the models, the material properties used in the FE model, and the loading conditions (i.e., external forces and constraints). This study indicates that FE models and modeling of skulls as simple engineering structures may give a preliminary idea of how these structures are loaded, but whenever possible, modeling results should be verified with either in vitro or preferably in vivo testing, especially if precise knowledge of strain magnitudes is desired. PMID- 15747348 TI - Mechanical behavior and quantitative morphology of the equine laminar junction. AB - The horse's hoof is structurally modified for its mechanical functions, but studying the functional design of internal structures is hampered by the external keratinous capsule. Finite-element analysis offers one method for evaluating mechanical function of components within the capsule, such as the laminar junction. This is the epidermodermal connection that binds the hoof wall strongly to the distal phalanx. Primary epidermal laminae (PEL), projecting inward from the wall, vary in morphology and are remodeled despite being keratinous. The aim of this study is to investigate the suggestion that remodeling of PEL is influenced by mechanical stress. Circumferential and proximodistal stress distribution and relative displacement in the laminar junction are assessed by finite-element analysis (FEA) of nine hoof models. Spacing, orientation, and curvature of PEL are assessed from sections through 47 other hooves and compared with the stress and displacement data. Significant correlations are found between laminar spacing and seven displacement and stress variables, supporting the link between stresses and remodeling. Differences in external hoof shape cause regional variation in stress magnitudes around the laminar junction. This finding is in accord with previous observations that laminar morphology is individually regionally variable. This work provides the first concrete link between mechanical behavior and laminar morphology. PMID- 15747349 TI - Australopithecus anamensis: a finite-element approach to studying the functional adaptations of extinct hominins. AB - Australopithecus anamensis is the stem species of all later hominins and exhibits the suite of characters traditionally associated with hominins, i.e., bipedal locomotion when on the ground, canine reduction, and thick-enameled teeth. The functional consequences of its thick enamel are, however, unclear. Without appropriate structural reinforcement, these thick-enameled teeth may be prone to failure. This article investigates the mechanical behavior of A. anamensis enamel and represents the first in a series that will attempt to determine the functional adaptations of hominin teeth. First, the microstructural arrangement of enamel prisms in A. anamensis teeth was reconstructed using recently developed software and was compared with that of extant hominoids. Second, a finite-element model of a block of enamel containing one cycle of prism deviation was reconstructed for Homo, Pan, Gorilla, and A. anamensis and the behavior of these tissues under compressive stress was determined. Despite similarities in enamel microstructure between A. anamensis and the African great apes, the structural arrangement of prismatic enamel in A. anamensis appears to be more effective in load dissipation under these compressive loads. The findings may imply that this hominin species was well adapted to puncture crushing and are in some respects contrary to expectations based on macromorphology of teeth. Taking together, information obtained from both finite-element analyses and dental macroanatomy leads us to suggest that A. anamensis was probably adapted for habitually consuming a hard-tough diet. However, additional tests are needed to understand the functional adaptations of A. anamensis teeth fully. PMID- 15747350 TI - Finite-element analysis of biting behavior and bone stress in the facial skeletons of bats. AB - The wide range of dietary niches filled by modern mammals is reflected in morphological diversity of the feeding apparatus. Despite volumes of data on the biomechanics of feeding, the extent to which the shape of mammal skulls reflects stresses generated by feeding is still unknown. In addition to the feeding apparatus, the skull accommodates the structural needs of the sensory systems and brain. We turned to bats as a model system for separating optimization for masticatory loads from optimization for other functions. Because the energetic cost of flight increases with body mass, it is reasonable to suggest that bats have experienced selective pressure over evolutionary time to minimize mass. Therefore, the skulls of bats are likely to be optimized to meet functional demands. We investigate the hypothesis that there is a biomechanical link between biting style and craniofacial morphology by combining biting behavior and bite force data gathered in the field with finite-element (FE) analysis. Our FE experiments compared patterns of stress in the craniofacial skeletons within and between two species of bats (Artibeus jamaicensis and Cynopterus brachyotis) under routine and atypical loading conditions. For both species, routine loading produced low stresses in most of the skull. However, the skull of Artibeus was most resistant to loads applied via its typical biting style, suggesting a mechanical link between routine loading and skull form. The same was not true of Cynopterus, where factors other than feeding appear to have had a more significant impact on craniofacial morphology. PMID- 15747351 TI - Modeling masticatory muscle force in finite element analysis: sensitivity analysis using principal coordinates analysis. AB - Our work on a finite element model of the skull of Macaca aims to investigate the functional significance of specific features of primate skulls and to determine to which of the input variables (elastic properties, muscle forces) the model behavior is most sensitive. Estimates of muscle forces acting on the model are derived from estimates of physiological cross-sectional areas (PCSAs) of the jaw muscles scaled by relative electromyographic (EMG) amplitudes recorded in vivo. In this study, the behavior of the model was measured under different assumptions regarding the PCSAs of the jaw muscles and the latency between EMG activity in those muscles and the resulting force production. Thirty-six different loading regimes were applied to the model using four different PCSA sets and nine different PCSA scaling parameters. The four PCSA sets were derived from three different macaque species and one genus average, and the scaling parameters were either EMGs from 10, 20, 30, 40, 50 and 60 msec prior to peak bite force, or simply 100%, 50%, or 25% of peak muscle force. Principal coordinates analysis was used to compare the deformations of the model produced by the 36 loading regimes. Strain data from selected sites on the model were also compared with in vivo bone strain data. The results revealed that when varying the external muscle forces within these boundaries, the majority of the variation in model behavior is attributable to variation in the overall magnitude rather than the relative amount of muscle force generated by each muscle. Once this magnitude-related variation in model deformation was accounted for, significant variation was attributable to differences in relative muscle recruitment between working and balancing sides. Strain orientations at selected sites showed little variation across loading experiments compared with variation documented in vivo. These data suggest that in order to create an accurate and valid finite element model of the behavior of the primate skull at a particular instant during feeding, it is important to include estimates of the relative recruitment levels of the masticatory muscles. However, a lot can be learned about patterns of skull deformation, in fossil species for example, by applying external forces proportional to the estimated relative PCSAs of the jaw adductors. PMID- 15747352 TI - Finite-element modeling of the anthropoid mandible: the effects of altered boundary conditions. AB - Finite-element modeling provides a full-field method for describing the stress environment of the skull. The utility of finite-element models, however, remains uncertain given our ignorance of whether such models validly portray states of stress and strain. For example, the effects of boundary conditions that are chosen to represent the mechanical environment in vivo are largely unknown. We conducted an in vitro strain gauge experiment on a fresh, fully dentate adult mandible of Macaca fascicularis to model a simplified loading regime by finite element analysis for purposes of model validation. Under various conditions of material and structural complexity, we constructed dentate and edentulous models to measure the effects of changing boundary conditions (force orientation and nodal constraints) on strain values predicted at the gauge location. Our results offer a prospective assessment of the difficulties encountered when attempting to validate finite-element models from in vivo strain data. Small errors in the direction of load application produce significant changes in predicted strains. An isotropic model, although convenient, shows poor agreement with experimental strains, while a heterogeneous orthotropic model predicts strains that are more congruent with these data. Most significantly, we find that an edentulous model performs better than a dentate one in recreating the experimental strains. While this result is undoubtedly tied to our failure to model the periodontal ligament, we interpret the finding to mean that in the absence of occlusal loads, teeth within alveoli do not contribute significantly to the structural stiffness of the mandible. PMID- 15747353 TI - Levodopa-responsive infantile parkinsonism due to a novel mutation in the tyrosine hydroxylase gene and exacerbation by viral infections. AB - Autosomal recessive forms of infantile dystonia due to mutations in the tyrosine hydroxylase (TH) gene have been described recently. The main clinical manifestations are Segawa's disease, or infantile hypokinetic rigid Parkinsonism. Here, we report on a patient with hyperrigidity, psychomotor developmental delay, and dystonic posturing of the hands, symptoms that appeared after a viral infection at the age of 14 months. Low homovanillic acid/5-hydroxyindolacetic acid (HVA/5HIAA) ratio in cerebrospinal fluid suggested a TH deficiency. Molecular analysis revealed a novel (H246Y) and a known (D498G) compound heterozygote mutation in the TH gene. The patient showed a remarkable response to treatment with levodopa. The new mutation and the association of viral infections with the onset and worsening of symptoms are discussed. PMID- 15747355 TI - Finite element analysis in functional morphology. AB - This article reviews the fundamental principles of the finite element method and the three basic steps (model creation, solution, and validation and interpretation) involved in using it to examine structural mechanics. Validation is a critical step in the analysis, without which researchers cannot evaluate the extent to which the model represents or is relevant to the real biological condition. We discuss the method's considerable potential as a tool to test biomechanical hypotheses, and major hurdles involved in doing so reliably, from the perspective of researchers interested in functional morphology and paleontology. We conclude with a case study to illustrate how researchers deal with many of the factors and assumptions involved in finite element analysis. PMID- 15747354 TI - Serum from Sydenham's chorea patients modifies intracellular calcium levels in PC12 cells by a complement-independent mechanism. AB - The proposed pathogenesis of Sydenham's chorea (SC) is an autoantibody-mediated basal ganglia dysfunction. Our study has shown that incubation of PC12 cells with complement-inactivated serum from SC patients was associated with a significant increase in Ca2+ levels evoked by KCl stimulus (mean +/- SEM, 341.0 +/- 8.7% of fluorescence intensity, arbitrary units) when compared with incubation with control serum (313.8 +/- 8.7% of fluorescence intensity, arbitrary units; P = 0.01). The increase in Ca2+ levels determined by SC patients sera correlated directly with the enzyme-linked immunosorbent assay optical density values for anti-basal ganglia antibodies. Our study supports the hypothesis that antibodies against basal ganglia in SC may cause their dysfunction. PMID- 15747357 TI - Genetic polymorphism of catechol-O-methyltransferase and levodopa pharmacokinetic pharmacodynamic pattern in patients with Parkinson's disease. AB - We explored the potential effect of catechol-O-methyltransferase (COMT) genetic polymorphism on the pharmacokinetics and pharmacodynamics of a standard oral dose of levodopa in patients with Parkinson's disease (PD). We prospectively collected blood samples for COMT genotyping from a population of 104 PD patients. Each patient was examined by a standard oral levodopa/benserazide test, based on simultaneous serial measurements of plasma levodopa concentrations, finger tapping motor effects and dyskinesia ratings, up to 4 hours after dosing. The main levodopa pharmacokinetic outcome variables were time to peak and peak plasma concentration, plasma elimination half-life, and the area under the plasma concentration-time curve. The main outcome levodopa pharmacodynamic variables were latency, duration, and magnitude of the motor effect elicited by the levodopa test dose, the area under the tapping effect-time curve, and the presence of dyskinesias. Nineteen patients (18%) harbored the low-activity homozygous COMT genotype (A/A), 63 patients (61%) carried the intermediate activity heterozygous COMT genotype (A/G) and 22 patients (21%) had the high activity homozygous COMT genotype (G/G). The three groups were comparable for vital and clinical characteristics. No significant difference was found in levodopa main pharmacokinetic-pharmacodynamic variables and dyskinesia incidence among the three subgroups of patients. We failed to identify clinically relevant levodopa pharmacokinetic-pharmacodynamic response patterns associated with the COMT polymorphism in PD patients. PMID- 15747356 TI - Familial cortical myoclonic tremor with epilepsy: a single syndromic classification for a group of pedigrees bearing common features. AB - Fifty Japanese and European families with cortical myoclonic tremor and epilepsy have been reported under various names. Unfamiliarity with the syndrome often leads to an initial misdiagnosis of essential tremor or progressive myoclonus epilepsy. A detailed overview of the literature is lacking and is the scope of this study. Disease characteristics are adult onset, distal action tremor and myoclonus, epileptic seizures, autosomal dominant inheritance, benign course, effectiveness of antiepileptic drugs, and possibly cognitive decline. A channelopathy is hypothesized to be the basis of the disease. Despite phenotypic and genetic differences between the Japanese and European pedigrees, the clinical and electrophysiological data point toward one syndrome. To avoid confusion in literature and possible misdiagnosis of patients, we propose to use one description and suggest "familial cortical myoclonic tremor with epilepsy" (FCMTE). In addition, we put forward diagnostic criteria to give a starting point from which to conduct genetic studies. PMID- 15747358 TI - Clinical grand round: a rapidly progressive pyramidal and extrapyramidal syndrome with a supranuclear gaze palsy. AB - The case history of a patient with a rapidly progressive spastic tetraparesis, pseudobulbar palsy, supranuclear gaze palsy, and extrapyramidal signs is presented. The patient died 17 months after symptom onset, and the differential diagnosis and pathological findings are reviewed. PMID- 15747359 TI - Performance measures in Friedreich ataxia: potential utility as clinical outcome tools. AB - Although several neuroprotective agents have been proposed as potential therapies in Friedreich ataxia (FA), clinical trials of their efficacy are limited by a lack of sensitive outcome measures. We assessed whether performance measures (nine-hole peg test, the timed 25-foot walk, and low-contrast letter acuity) provide valid measures of disease status in FA. Scores for each measure correlated significantly with neurologic disability and disease duration. Rank correlations between scores for performance measures were moderate in magnitude, suggesting that the each test captures separate yet related dimensions of neurological function in FA. Linear regression models demonstrated that scores from the nine-hole peg test and the timed 25-foot walk (after reciprocal transformation) were predicted by age and triplet repeat length in patients with FA. In addition, comparison of the temporal courses of change for each performance measure demonstrated that scores from the timed 25-foot walk change early in the course of FA, nine-hole peg test scores change slowly over the full course of the disorder, and low-contrast letter acuity scores change in the later stages of the disease. Thus, a composite scale derived from these performance measures may provide the best overall measure for assessing disease progression throughout the illness. PMID- 15747360 TI - Novel compound heterozygous mutations in the PANK2 gene in a Chinese patient with atypical pantothenate kinase-associated neurodegeneration. AB - We investigated the presence of mutations in the pantothenate kinase (PANK2) gene in a 27-year-old male Chinese patient with atypical pantothenate kinase associated neurodegeneration (PKAN), formerly Hallervorden-Spatz syndrome. Automated DNA sequence analyses revealed compound heterozygous mutations in the exon 3 and 5. This patient had a 10-year history of PKAN characterized by a slight tremor of the right hand when writing at onset and a slow progressive rigidity of the neck and the right arm and resting tremor in upper extremities. Dysarthria, dysphagia, and dystonic-athetoid movements of the face and right fingers were marked. Magnetic resonance showed the typical "eye-of-the-tiger" sign. PMID- 15747362 TI - Soleus H-reflex inhibition during gait initiation in Parkinson's disease. AB - The soleus H-reflex excitability during gait initiation was investigated in Parkinson's disease. Eleven patients participated in this study. Patients stepped forward as soon as the start signal flashed. Soleus H-reflex was evoked from the trailing leg 100, 300, or 600 msec after the start signal. The electromyographic activity in the soleus muscle immediately before evoking the H-reflex and the ankle joint motion were recorded. The soleus H-reflex was inhibited 300 msec after the start signal. The amount of the soleus H-reflex inhibition was inversely correlated with the Hoehn and Yahr stage; Items 14, 29, and 31 of the Unified Parkinson's Disease Rating Scale; and the delay of the onset of the ankle dorsiflexion from the start signal. In contrast, the amount of electromyographic activity immediately before evoking the H-reflex was not significantly correlated with those measures but was significantly correlated with Item 22 of the Unified Parkinson's Disease Rating Scale. Those findings indicate that the amount of soleus H-reflex inhibition during gait initiation depends on the severity of the disease. Abnormality of descending command may be related to the severity dependent H-reflex inhibition. PMID- 15747361 TI - Botulinum toxin type A for treatment of spasticity in spinocerebellar ataxia type 3 (Machado-Joseph disease). PMID- 15747363 TI - Botulinum toxin and skin rash reaction. PMID- 15747364 TI - A microscope on vertebrate form and function: the power of finite element analysis. PMID- 15747366 TI - Does cigarette smoking provide clinically significant neuroprotection among patients diagnosed with Parkinson's disease? PMID- 15747367 TI - The neurochemical mechanism of rebound psychosis in Parkinson's disease. PMID- 15747368 TI - Chorea in a patient with cerebral palsy: treatment with levetiracetam. AB - We report on the case of an adult cerebral palsy patient who developed severe chorea coincident with a febrile illness from a nonstreptococcal infection. The chorea improved markedly with the use of levetiracetam (LEV, Keppra). PMID- 15747369 TI - Impact of internal versus external cueing on driving performance in people with Parkinson's disease. AB - Numerous aspects of driving performance seem compromised in people with Parkinson's disease (PD). Measures of cognitive impairment consistently correlate with poor driving simulator performance in this population; however, the effects of specific cognitive difficulties on discrete aspects of driving behavior have not been investigated thoroughly. Previous studies have demonstrated that people with PD exhibit difficulties internally cueing cognitive processes. This study examined the impact of impaired internal cueing on specific driving behaviors. A simulator measured the driving behavior of 18 current drivers in the mild-to moderate stages of PD and 18 matched controls. Participants navigated through different driving conditions where the opportunity to use internal and external cues was manipulated. People with PD exhibited difficulties using internal cues to regulate driving behavior around traffic signals and curves. Instead of using internal cues, participants with PD were more reliant on external cues to regulate driving behavior. They were also less able to adapt their driving behavior to suit driving conditions. Because all participants with PD were current drivers in the mild-to-moderate stages of the disease, findings challenge the widely-held assumption that cognitive difficulties only impact on driving performance in the moderate-to-severe stages of PD. PMID- 15747370 TI - Reversible withdrawal dyskinesia associated with quetiapine. PMID- 15747371 TI - Age at onset variance analysis in spinocerebellar ataxias: a study in a Dutch French cohort. AB - In dominant spinocerebellar ataxias (SCAs), the issue of whether non-CAG dependent factors contribute to onset age remains unsettled. Data on SCA genotype, onset age, normal/expanded CAG repeat length, sex of the patient and transmitting parent, and family details were available from 802 patients. Based on the model [log(10) (age at onset) = k - b CAG(exp) + epsilon], we examined changes in adjusted R(2) and residual standard error following incorporation of the other factors in this model. The expanded repeat explained 44.3 to 74.9% of onset age variance, although this was less than 50% in SCA3 and SCA6, implicating a large effect of non-CAG factors. The relation between onset age and CAG repeat was similar for SCA1, 3, 6, and 7, but different for SCA2, pointing to different polyglutamine effects in SCA2. For SCA2 and SCA3, 17.1 and 45.5% of onset age variance, respectively, were explained by currently (unidentified) familial factors. We found a significant contribution of the nonexpanded allele in SCA1 and SCA6. Besides polyglutamine motif (determined by the expanded CAG repeat length), we identified the following age at onset modifiers: protein context in SCA2; familial factors in SCA2 and SCA3; and the nonexpanded CAG repeat in SCA1 and SCA6. PMID- 15747372 TI - Non-movement disorder heralds symptoms of Machado-Joseph disease years before ataxia. AB - We describe three patients with the Machado-Joseph disease (MJD) genetic abnormality who had non-movement disorder neurological symptoms or signs that preceded the gait ataxia by several years. This implies that some clinical manifestations other than ataxia may be considered part of the herald symptoms of MJD, especially in the setting of a positive family history. PMID- 15747373 TI - Capillary electrophoresis-mass spectrometry as a powerful tool in clinical diagnosis and biomarker discovery. AB - Proteome analysis is now emerging as key technology for deciphering biological processes and the discovery of biomarkers for diseases from tissues and body fluids. The complexity and wide dynamic range of protein expression poses a formidable challenge to both peptide separation technologies and mass spectrometry (MS). Here we review the efforts that have been undertaken to date, focussing on capillary electrophoresis coupled to mass spectrometry (CE-MS). We discuss CE-MS from an application point of view evaluating its merits and vices in regard to biomarker discovery and clinical applications. As examples, we present the use of CE-MS for the determination of protein patterns in urine, serum, and other body fluids. Finally, the benefits and limitations of CE-MS for the analysis of proteins in clinical samples are discussed against the background of alternative technologies. PMID- 15747374 TI - Cryosurgical ablation of the prostate: high risk patient outcomes. AB - BACKGROUND: The authors report their experience with cryosurgical ablation of the prostate in men with high-risk features for prostate carcinoma who were unwilling to undergo radical surgery or radiation therapy. METHODS: Between January 1998 and April 2002, 65 men underwent primary cryosurgery for prostate carcinoma with high-risk features. All patients had biopsy-proven prostate carcinoma without evidence for metastatic disease on magnetic resonance images, computed tomography scans, or radionuclide images of bones. High-risk parameters were defined as either a prostate-specific antigen (PSA) level >/= 10 ng/mL, or a Gleason sum score >/= 8, or both. Patients who had undergone prior surgery, radiation therapy, or cryoablation for prostate carcinoma were excluded from the study. Patients were monitored with physical examination and PSA screening every 3 months and with radiologic imaging when indicated. RESULTS: The median patient age was 72 years (range, 41-86 years), and t he median follow-up was 35 months (range, 4-77 months). There were 2 patients (3.1%) with rectal pain and incontinence. Durable PSA biochemical disease-free survival was noted in 83.3% of patients according to the American Society for Therapeutic Radiology and Oncology (ASTRO) criteria. A 6-year Kaplan-Meier analysis revealed an 81.7% ASTRO survival probability as well as PSA nadir < 4.0 ng/mL and PSA nadir < 1.0 ng/mL projections of 50% and 35%, respectively. One of 8 postcryosurgery biopsies (12.5%) were positive. No patient had progressed at last follow-up, and the overall survival rate was 100%. CONCLUSIONS: Cryoablation was a feasible treatment option in patients with organ-confined prostate carcinoma who had high risk features. Longer follow-up will be necessary to determine the effectiveness of this approach. PMID- 15747375 TI - Clinical outcome of patients with lymph node-negative breast carcinoma who have sentinel lymph node micrometastases detected by immunohistochemistry. AB - BACKGROUND: The ideal pathologic assessment of sentinel lymph nodes (SLNs) in patients with breast carcinoma remains controversial. The authors evaluated how detailed assessment of SLNs using immunohistochemistry (IHC) and serial sectioning would affect treatment decisions and outcomes in patients with breast carcinoma who had negative SLNs on standard hematoxylin and eosin staining. METHODS: The SLNs from patients who were treated between June 1998 and June, 1999 and who had negative lymph node status determined by hematoxylin and eosin staining (n = 84 patients) were evaluated further with serial sectioning and cytokeratin IHC. Patients were offered adjuvant therapy based on primary tumor factors. RESULTS: The median patient age was 57 years, and the median tumor size was 1.2 cm. At a median follow-up of 40.2 months, 81 patients (96%) were alive with no evidence of disease, 1 patient was alive with disease, 1 patient had died of disease, and 1 patient had died of other causes. Fifteen patients (18%) had micrometastases identified on IHC. Of the total 84 patients, information regarding adjuvant therapy was not available for 5 patients. Of the remaining 79 patients, 10 patients (13%) were not offered adjuvant chemotherapy but had positive SLN status determined by IHC. SLN status based on IHC evaluation did not correlate with age (P = 0.077), tumor size (P = 0.717), grade (P = 0.148), estrogen receptor status (P = 1.000), or lymphovascular invasion (P = 0.274). Furthermore, IHC-detected positive SLN status did not correlate with distant metastasis (P = 0.372) or overall or distant metastasis-free survival (P = 0.543 and P = 0.540, respectively). CONCLUSIONS: Although the finding of SLN micrometastases by IHC may change management in > 12% of patients, preliminary results suggested that such micrometastases do not affect outcomes significantly. PMID- 15747376 TI - Response and resistance in 300 patients with BCR-ABL-positive leukemias treated with imatinib in a single center: a 4.5-year follow-up. AB - BACKGROUND: The advent of imatinib has considerably changed the treatment of chronic myeloid leukemia (CML). Early studies demonstrated high rates of hematologic and cytogenetic responses in all phases of the disease after limited observation periods. METHODS: The authors evaluated long-term outcome, rates of response, and resistance in 300 patients with BCR-ABL-positive leukemias (CML in chronic phase after failure to respond to interferon-alpha [CP], n = 139; accelerated phase [AP], n = 80; myeloid blast crisis [BC], n = 76; lymphoid BC and Philadelphia chromosome-positive acute lymphoblastic leukemia, n = 5) who entered clinical trials with imatinib in a single center after an observation time of 4.5 years. RESULTS: In CP, hematologic remission was achieved in 97% and major (MCR) and complete cytogenetic remission (CCR) in 61% and 49% of patients, respectively. The chance to achieve MCR was higher in patients commencing imatinib earlier in the course of CML. In AP, the median survival period after the start of imatinib was 44 months, and MCR and CCR were observed in 31% and 26% of patients, respectively. In myeloid BC, the median survival period after the start of imatinib and after diagnosis of BC was 6 and 9 months, respectively. Hematologic resistance occurred in 25%, 41%, and 92% of patients in CP, AP, and myeloid BC, respectively, and was associated with BCR-ABL mutations in 45% of patients and with clonal evolution in 58% of patients. CONCLUSIONS: The data emphasized the need for a prolonged follow-up of patients treated with imatinib to define the clinical potential of the drug and to establish methods to optimize therapy. PMID- 15747377 TI - Effect of sulphur mustard on human skin cell lines with differential agent sensitivity. AB - The ability of sulphur mustard (HD) to induce DNA damage places limits on the efficacy of approaches aimed at protecting human cells from the cytotoxic effects of HD using a variety of protective agents such as thiol-containing esters and protease inhibitors. In the present study, potential alternative strategies were investigated by examining the differential effects of HD on G361, SVK14, HaCaT and NCTC 2544 human skin cells. The G361 cell line was more resistant to the cytotoxic effects of HD than the NCTC, HaCaT and SVK14 cell lines at HD doses of >3 and <100 microM HD as determined by the MTT assay. At 72 h after exposure to 60 microM HD there was up to an 8.8-fold difference (P < 0.0001) between G361 and SVK14 cell culture viability. Buthionine sulphoximine (BSO) pretreatment increased the sensitivity of all four cell lines to HD. A substantial proportion of the resistance of G361 cells to HD was attributable to BSO-mediated effects on antioxidant-mediated metabolism, although G361 cultures still retained a high degree of viability at 30 microM HD following BSO pretreatment. Cell cycle analysis confirmed that SVK14 cells were relatively more sensitive to HD, as shown by the 2.1-fold reduction (P < 0.0001) in the percentage of cells in G0/G1 phase 24 h after HD exposure compared with control cultures. This compared well with a 1.2-fold increase (P < 0.05) in the percentage of G361 cells in G0/G1 phase following HD exposure, suggesting the existence of a more efficient G0/G1 checkpoint control mechanism in this cell line. Manipulation of the cell cycle using various modulating agents did not increase the resistance of cell lines to the cytotoxic effects of HD. PMID- 15747380 TI - Regeneration of rat auditory ossicles using recombinant human BMP-2/collagen composites. AB - Tympanoplasty operations to improve hearing impairment require a wide middle ear cavity and reconstruction of columellar formations. There is no specific material for use in the reconstruction of columellar formations. Tissue response to BMP has been employed as regenerative material. To our knowledge, however, there are no reports of the reconstruction of columellar formations using recombinant human bone morphogenetic protein-2/bovine collagen composites in the middle ear. The purpose of this study is to investigate whether recombinant human bone morphogenetic protein-2/bovine collagen composites (rhBMP-2 composites) are appropriate for use as regenerative material for tympanoplasty. In the form of pellets, rhBMP-2 composites were implanted as columellae into the tympanic cavity. At 2, 4, 6, 8, and 12 weeks after surgery, the middle ear of the animals (n = 4 at each week) was stained with hematoxylin-eosin for light microscopic observation. All composites were in the process of ossification or had ossified according to their developmental stages and were covered with a single layer of squamous or cuboidal epithelium. The new bone formed in these composites was persistently stable and displayed some columellar conditions assessed by histological examination. This study led to the conclusion that rhBMP-2 composites make excellent regenerative material for auditory ossicles. PMID- 15747378 TI - Involvement of chitin in exoskeleton morphogenesis in Drosophila melanogaster. AB - Exoskeletons stabilize cell, tissue, and body morphology in many living organisms including fungi, plants, and arthropods. In insects, the exoskeleton, the cuticle, is produced by epidermal cells as a protein extracellular matrix containing lipids and the polysaccharide chitin, and its formation requires coordinated synthesis, distribution, and modification of these components. Eventually, the stepwise secretion and sorting of the cuticle material results in a layered structure comprising the envelope, the proteinaceous epicuticle, and the chitinous procuticle. To study the role of chitin during cuticle development, we analyzed the consequences of chitin absence in the embryo of Drosophila melanogaster caused by mutations in the Chitin Synthase-1 (CS-1) gene, called krotzkopf verkehrt (kkv). Our histological data confirm that chitin is essential for procuticle integrity and further demonstrate that an intact procuticle is important to assemble and to stabilize the chitin-less epicuticle. Moreover, the phenotype of CS-1/kkv mutant embryos indicates that chitin is required to attach the cuticle to the epidermal cells, thereby maintaining epidermal morphology. Finally, sclerotization and pigmentation, which are the last steps in cuticle differentiation, are impaired in tissues lacking CS-1/kkv function, suggesting that proper cuticle structure is crucial for the activity of the underlying enzymes. PMID- 15747381 TI - Effects of filler composition on flexibility of microfilled resin composite. AB - The effects of the filler composition on physical and mechanical properties of microfilled composites was investigated by measuring water absorption, solubility, compressive, flexural, and impact strength. A series of experimental composites, consisting of UDMA/TEGDMA comonomer matrix and prepolymerized fillers, was fabricated. The prepolymerized fillers were composed of hydrophobic colloidal silica and two monomers in varying ratios, trimethylolpropanetrimethacrylate (TMPT), and polyesterdiacrylate (PEDA). TMPT/PEDA ratios were 100:0, 64:36, 46:54, 18:82, and 0:100%. There were no significant differences in water sorption and solubility, regardless of the amount of PEDA monomer. Young's modulus and modulus of resilience increased with decreasing PEDA ratio. Fracture energy exhibited drastic changes (30.1 x 10(-5) J to 93.4 x 10(-5) J). The highest value of flexural strength (96.0 +/- 3.5 MPa) was obtained when the TMPT-PEDA filler was 46:54. The impact strengths of composites fabricated with TMPT-PEDA filler of 46:54 (11.2 +/- 1.4 kJ/m(2)), 18:82 (10.6 +/- 3.2 kJ/m(2)), and 0:100 (13.1 +/- 3.8 kJ/m(2)) were significantly higher than those with 100:0 (6.0 +/- 1.8 kJ/m(2)) or 64:36 (7.1 +/- 2.4 kJ/m(2)). Based upon the results, it was concluded that the mechanical properties of microfilled composites were improved by the modification of prepolymerized filler composition. PMID- 15747382 TI - Are HMOs bad for health maintenance? AB - This study examines the impact of Health Maintenance Organization (HMO) coverage on the provision of preventive medicine. We investigate whether any association reflects selection effects on the part of patients and/or physicians or a causal impact of managed care itself. Causal effects may occur on the supply side or the demand side. Using a large national database of Medicare and non-Medicare patients, we investigate these issues for eight common preventive medical procedures. We find that preventive care is substantially higher with HMO coverage than with traditional fee-for-service reimbursement. Our findings also suggest that the impact of HMOs on preventive medicine is a causal one, and does not merely reflect selection effects. Both supply-side (e.g. provider) and demand side (e.g. patient) factors appear to play a role in the higher incidence of preventive care among HMO enrollees. Patient demand effects are stronger for simple treatments such as physicals, while supply-side effects seem to dominate for relatively complex preventive care procedures such as mammograms. PMID- 15747383 TI - Genetic and environmental influences on symptomatic gallstone disease: a Swedish study of 43,141 twin pairs. AB - The contribution of hereditary and environmental factors to the pathogenesis of symptomatic gallstone disease is still unclear. We estimated the relative importance of genetic and environmental factors by analyzing a large population of twins. For this purpose, the Swedish Twin Registry was linked with the Swedish inpatient-discharge and causes of death registries for symptomatic gallstone disease and gallstone surgery-related diagnoses in 43,141 twin pairs born between 1900 and 1958. Concordance rates, correlations, and odds ratios were calculated for males, females, monozygotic, and dizygotic twins, respectively, as well as for twin pairs of opposite sex. Structural equation modeling techniques were used to estimate the contributions of genetic effects as well as shared and non-shared environmental factors to the development of symptomatic gallstone disease. We found that concordances and correlations were higher in monozygotic compared with dizygotic twins, both for males and females. Of note, there were no significant sex differences in heritability. In the full model, genetic effects accounted for 25% (95% CI, 9%-40%), shared environmental effects for 13% (95% CI, 1%-25%), and unique environmental effects for 62% (95% CI, 56%-68%) of the phenotypic variance among twins. In conclusion, our results show heritability to be a major susceptibility factor for symptomatic gallstone disease, consistent with results from previous, much smaller studies. PMID- 15747384 TI - Morphological and molecular analysis of the early developing chick requires an expanded series of primitive streak stages. AB - A detailed analysis of the gastrulating chick embryo was performed using three methods : time-lapse videotaping of embryos in culture, histological semithin sections, and in situ hybridization with 10 mRNA signals expressed during gastrulation. The results suggest that the gene expression pattern of Goosecoid, Hex, Crescent, and Bmp7 may be involved in the axial establishment of the temporal and spatial arrangement of cells forming the prechordal plate endoderm, and that Chordin, cNot1, Noggin, and Brachyury are precocious markers of cells coming from Hensen's node, which contribute to the rostralmost tip of the notochord, its arrowhead, the head process, and, later, the elongating notochord. These results explain several earlier descriptions based only on morphological analyses of the axial mesodermal structures characteristic of the gastrulation stages. The data, carefully observed and compared with the whole-mount observation in time-lapse video, show that the changes in cell populations, movements, and cell differentiation occur step-by-step over a precise temporal range, which requires the establishment of a subdivision of the stages usually employed. Knowledge of new aspects of avian gastrulation, including gene expression patterns, immunocytochemical analyses, and the great number of recent experiments based on microinjections or transplants of groups of cells to analyze processes of induction or regulation, need the support of a precisely defined scheme of primitive streak stages (PS-stages), and a correlation of these stages with other approaches to provide a finer resolution of the staging steps, and thus to facilitate a better understanding of the initial gastrulation period. PMID- 15747385 TI - A cobaltocenium-peptide bioconjugate shows enhanced cellular uptake and directed nuclear delivery. PMID- 15747386 TI - Convergent diversity-oriented synthesis of small-molecule hybrids. PMID- 15747387 TI - Calculation of the redox potential of the protein azurin and some mutants. AB - Azurin from Pseudomonas aeruginosa is a small 128-residue, copper-containing protein. Its redox potential can be modified by mutating the protein. Free-energy calculations based on classical molecular-dynamics simulations of the protein and from mutants in aqueous solution at different pH values were used to compute relative redox potentials. The precision of the free-energy calculations with the lambda coupling-parameter approach is evaluated as function of the number and sequence of lambda values, the sampling time and initial conditions. It is found that the precision is critically dependent on the relaxation of hydrogen-bonding networks when changing the atomic-charge distribution due to a change of redox state or pH value. The errors in the free energies range from 1 to 10 k(B)T, depending on the type of process. Only qualitative estimates of the change in redox potential by protein mutation can be obtained. PMID- 15747388 TI - Direct and stereoselective synthesis of 2-acetamido-2-deoxy-beta-D glycopyranosides by using the phosphite method. PMID- 15747389 TI - Unmyelinated fiber sensory neuropathy differs in type 1 and type 2 diabetes. AB - BACKGROUND: Neuropathic pain is common in diabetic patients. Degeneration of sensory C-fibers in peripheral nerve plays a prominent role in the generation of neuropathic pain. We examined degenerative changes of C-fibers in two rat models with type 1 and type 2 diabetes. METHODS: Type 1 insulinopenic BB/Wor and type 2 hyperinsulinemic diabetic BBZDR/Wor-rats of 8 months duration with equal exposure to hyperglycemia were examined. Thermal hyperalgesia was monitored using an infrared thermal probe. C-fiber size, number, frequencies of denervated Schwann cells, regenerating C-fibers, type 2 axon/Schwann cell relationship and collagen pockets in the sural nerve were examined morphometrically. Neurotrophic receptor expression was examined by Western blotting. Neurotrophins and neuropeptides were examined by ELISA. RESULTS: Type 1 rats showed increased thermal hyperalgesia followed by a decrease. Hyperalgesia in type 2 rats showed a slower progression. These findings were associated with a 50% (p < 0.001) loss of C-fibers, increased frequencies of denervated Schwann cells (p < 0.001), regenerating fibers (p < 0.001), collagen pockets (p < 0.001) and type 2 axon/Schwann cell relationship (p < 0.001) in type 1, but not in type 2 rats. Expression of insulin receptor, IGF 1R, TrkA and C was decreased in BB/Wor rats, whereas BBZDR/Wor rats showed milder or no deficits. NGF and NT-3 in sciatic nerve and substance P and calcitonin gene related peptide in dorsal root ganglia were decreased in type 1, but not in type 2 rats. CONCLUSION: The more severe molecular, functional and morphometric abnormalities of nociceptive C-fibers in type 1 insulinopenic rats compared to type 2 hyperinsulinemic rats suggest that impaired insulin action may play a more important pathogenetic role than hyperglycemia per se. PMID- 15747391 TI - Proceedings of the First Annual Conference of the MidSouth Computational Biology and Bioinformatics Society. November 13-15, 2003. Little Rock, Arkansas, USA. PMID- 15747390 TI - Toxin-based selection of insulin-producing cells with improved defense properties for islet cell transplantation. AB - Insulin-producing pancreatic beta-cells are known to be extremely susceptible to destruction, primarily by autoimmune mechanisms, infectious agents, and by chemical toxins that cause overt type I diabetes. As development of highly protected insulin-producing cells would be important for successful cell therapy of diabetic patients, gene transfection technique was utilized by several investigators in order to improve the defense properties of transplanted cells. In this article, we summarize other approaches based on a selection strategy that has been developed in our laboratory and by other research groups that engineer pancreatic beta-cells to provide protection against diabetogenic toxins (streptozotocin and alloxan), oxidative stress and cytokines. Selection strategies based on acute repeated or long-term continuous treatment of cell lines with cytotoxic agents have resulted in the selection of highly resistant cell subpopulations. We discuss possible involvement of different expression of cytoprotective genes in the selection of cell subpopulations, which demonstrate a broad spectrum of resistance. Importantly, toxin-based selection did not impair functional activity of the cells as it was shown in vitro. In addition, selected cells preserved their improved metabolic characteristics following encapsulation in alginate and subsequent implantation in diabetic animals. Identifying the mechanisms through which cell defense properties act will help clarify the process responsible for beta-cell regeneration in type I diabetes patients. Such knowledge might be useful in developing strategies focusing on the regeneration of beta-cell resistant populations. PMID- 15747392 TI - American Academy of Allergy, Asthma and Immunology (AAAAI) 61st annual meeting. San Antonio, Texas, USA, March 18-22, 2005. Abstracts. PMID- 15747393 TI - Rhabdomyosarcoma. PMID- 15747394 TI - Biological and physical dosimetry for radiation protection. Proceedings of a symposium. March 10, 2004. Braunschweig, Germany. PMID- 15747395 TI - More ZAP for chronic lymphocytic leukemia (CLL). AB - Antigen stimulation of the leukemic clone is increasingly implicated in the pathogenesis of CLL. Chen and colleagues provide evidence that expression of ZAP 70 in CLL B cells renders IgM signaling more effective and thereby could contribute to the more rapidly progressive clinical course of ZAP-70-positive CLL. PMID- 15747396 TI - Can we afford to let sleeping dogs lie? AB - In patients with chronic myelogenous leukemia (CML) mutations of the BCR-ABL kinase domain (KD) have been identified as the leading cause of acquired resistance to imatinib, while the mechanisms underlying the persistence of minimal residual disease (MRD) are unknown. In this issue of Blood, Chu and colleagues report several patients with KD mutations at the time of complete cytogenetic response (CCR), implicating mutations as a cause of disease persistence. PMID- 15747397 TI - "T"-ing off on nasopharynx cancer. AB - Nasopharyngeal carcinoma (NPC) is the most frequent Epstein-Barr virus (EBV) associated malignancy worldwide. In this issue, Straathof and colleagues take aim at EBV-associated NPC showing that adoptive immunotherapy with EBV-specific T cells expanded in vitro can be successfully administered to NPC patients and is associated with potential therapeutic efficacy. PMID- 15747398 TI - HIV-associated lymphoma: promising new results, but with toxicity. AB - Treatment of HIV-associated non-Hodgkin lymphoma with rituximab plus infusional cyclophosphamide, doxorubicin, and etoposide resulted in high complete remission and 2-year failure free and overall survival rates but a high rate of infection. PMID- 15747399 TI - Profiling is a good thing (at least in the clinic). AB - Monti and colleagues take the next step in the molecular characterization of diffuse large B-cell lymphoma (DLBCL). PMID- 15747400 TI - "You break it, you fix it.". AB - Nonrandom chromosomal translocations are considered to be causal events leading to leukemic transformation. However, the mechanism(s) that cause these translocations are poorly understood. Libura and colleagues report exciting new findings on the mechanism(s) that lead to chromosomal translocation. PMID- 15747401 TI - Cellular iron metabolism: mitochondria in the spotlight. AB - New insights into cellular iron metabolism have been provided by the recognition that certain diseases are associated with mitochondrial iron overload and by the discovery of mitochondrial ferritin (MtFt) and mitochondrial iron transporters. PMID- 15747402 TI - Annexin A2: better left alone. AB - Zhang and McCrae demonstrate that APLA/b2GPI-mediated endothelial cell activation occurs via dimerization of annexin A2 molecules on the cell surface. PMID- 15747403 TI - NO role in EPC function. AB - The nitric oxide pathway plays an important role in modulating endothelial progenitor cell function, playing a critical role in blood vessel repair in response to injury. PMID- 15747404 TI - Soluble urokinase activator receptor (suPAR) in stem cell mobilization. AB - Selleri et al demonstrate that healthy donors given granulocyte-colony stimulating factor (G-CSF) have increases in plasma soluble plasminogen activator receptor (suPAR) levels, which in turn induces chemotaxis of CD34 cells. Selleri et al's findings help better define the complicated mechanism of stem cell mobilization by G-CSF and point to a wide role for uPAR in cell surface adhesion and recognition. PMID- 15747405 TI - Cytochrome c-dimyristoylphosphatidylglycerol interactions studied by asymmetrical flow field-flow fractionation. AB - Lipid membranes are well recognized ligands that bind peripheral and integral proteins in a specific manner and regulate their function. Cytochrome c (cyt c) is one of the partner peripheral protein that binds to the lipid membranes via electrostatic and hydrophobic interactions. In this study, asymmetrical flow field-flow fractionation (AsFlFFF) was used to compare the interactions of cyt c with the acidic phospholipid 1,2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol (DMPG), oleic acid (OA), and sodium dodecyl sulfate (SDS). The influence of pH and the cyt c-lipid molar mass ratios were evaluated by monitoring the diffusion coefficients and particle diameter distributions obtained for the free and lipid bound protein. The hydrodynamic particle diameter of cyt c (pI 10) was 4.1 nm at pH 11.4 and around 4.2 nm at pH 7.0 and 8.0. Standard molar mass marker proteins were used for calibration to obtain the molar masses of free cyt c and its complexes with lipids. AsFlFFF revealed the binding of cyt c to DMPG and to OA to be mainly electrostatic. In the absence of electrostatic interactions, minor complex formation occurred, possibly due to the extended lipid anchorage involving the hydrophobic cavity of cyt c and the hydrocarbon chains of DMPG or SDS. The possibility of the formation of the molten globule state of cyt c, induced by the interaction between cyt c and lipids, is discussed. PMID- 15747406 TI - Shellfish conflict in Baynes Sound: a strategic perspective. AB - The shellfish aquaculture industry (SAI) has operated in Baynes Sound, British Columbia (BC) since the early 1900s. Recognizing the economic potential of the area, the industry has requested additional farming opportunities. However, Baynes Sound upland residents and many other stakeholders have expressed concerns that SAI activities are having a negative impact on the environment, quality of life, and other nonaquaculture resource uses in the area. In order to address these issues, the Action Plan was initiated by a BC government interagency project team in November 2001. To assist in assessing the strategic aspects of this conflict, the decision support system GMCR II is employed here to apply a new methodology, the graph model for conflict resolution, to systematically analyze the ongoing conflict over shellfish aquaculture development in Baynes Sound within a social, economic, and environmental framework. Valuable insights are procured to guide decision-makers toward sustainability of the shellfish industry. PMID- 15747407 TI - Hydrology or floristics? Mapping and classification of wetlands in Victoria, Australia, and implications for conservation planning. AB - A national approach to the conservation of biodiversity in Australia's freshwater ecosystems is a high priority. This requires a consistent and comprehensive system for the classification, inventory, and assessment of wetland ecosystems. This paper, using the State of Victoria as a case study, compares two classification systems that are commonly utilized to delineate and map wetlands- one based on hydrology (Victorian Wetland Database [VWD]) and one based on indigenous vegetation types and other natural features (Ecological Vegetation Classes [EVC]). We evaluated the extent of EVC mapping of wetlands relative to the VWD classification system using a number of datasets within a geographical information system. There were significant differences in the coverage of extant EVCs across bioregions, different-sized wetlands, and VWD wetland types. Resultant depletion levels were markedly different when examined using the two systems, with depletion levels, and therefore perceived conservation status, of EVCs being significantly higher. Although there is little doubt that many wetland ecosystems in Victoria are in fact threatened, the extent of this threat cannot accurately be determined by relying on the EVC mapping as it currently stands. The study highlighted the significant impact wetland classification methods have in determining the conservation status of freshwater ecosystems. PMID- 15747408 TI - Using wildlife as receptor species: a landscape approach to ecological risk assessment. AB - To assist risk assessors at the Department of Energy's Savannah River Site (SRS), a Geographic Information System (GIS) application was developed to provide relevant information about specific receptor species of resident wildlife that can be used for ecological risk assessment. Information was obtained from an extensive literature review of publications and reports on vertebrate- and contaminant-related research since 1954 and linked to a GIS. Although this GIS is a useful tool for risk assessors because the data quality is high, it does not describe the species' site-wide spatial distribution or life history, which may be crucial when developing a risk assessment. Specific receptor species on the SRS were modeled to provide an estimate of an overall distribution (probability of being in an area). Each model is a stand-alone tool consisting of algorithms independent of the GIS data layers to which it is applied and therefore is dynamic and will respond to changes such as habitat disturbances and natural succession. This paper describes this modeling process and demonstrates how these resource selection models can then be used to produce spatially explicit exposure estimates. This approach is a template for other large federal facilities to establish a framework for site-specific risk assessments that use wildlife species as endpoints. PMID- 15747409 TI - Rough set rule induction for suitability assessment. AB - The data that characterize an environmental system are a fundamental part of an environmental decision-support system. However, obtaining complete and consistent data sets for regional studies can be difficult. Data sets are often available only for small study areas within the region, whereas the data themselves contain uncertainty because of system complexity, differences in methodology, or data collection errors. This paper presents rough-set rule induction as one way to deal with data uncertainty while creating predictive if-then rules that generalize data values to the entire region. The approach is illustrated by determining the crop suitability of 14 crops for the agricultural soils of the Willamette River Basin, Oregon, USA. To implement this method, environmental and crop yield data were spatially related to individual soil units, forming the examples needed for the rule induction process. Next, four learning algorithms were defined by using different subsets of environmental attributes. ROSETTA, a software system for rough set analysis, was then used to generate rules using each algorithm. Cross-validation analysis showed that all crops had at least one algorithm with an accuracy rate greater than 68%. After selecting a preferred algorithm, the induced classifier was used to predict the crop suitability of each crop for the unclassified soils. The results suggest that rough set rule induction is a useful method for data generalization and suitability analysis. PMID- 15747410 TI - Hydrochemical buffer assessment in agricultural landscapes: from local to catchment scale. AB - Non-point-source pollution of surface and groundwater is a prominent environmental issue in rural catchments, with major consequences on water supply and aquatic ecosystem quality. Among surface-water protection measures, environmental or landscape management policies support the implementation and the management of buffer zones. Although a great number of studies have focused on buffer zones, quantification of the buffer effect is still a recurring question. The purpose of this article is a critical review of the assessment of buffer-zone functioning. Our objective is to provide land planners and managers with a set of variables to assess the limits and possibilities for quantifying buffer impact at the catchment scale. We first consider the scale of the local landscape feature. The most commonly used empirical method for assessing buffers is to calculate water/nutrient budgets from inflow-outflow monitoring at the level of landscape structures. We show that several other parameters apart from mean depletion of flux can be used to describe buffer functions. Such parameters include variability, with major implication for water management. We develop a theoretical framework to clarify the assessment of the buffer effect and propose a systematic analysis taking account of temporal variability. Second, we review the current assessment of buffer effects at the catchment scale according to the theoretical framework established at the local scale. Finally, we stress the limits of direct empirical assessment at the catchment scale and, in particular, we emphasize the hierarchy in hydrological processes involved at the catchment scale: The landscape feature function is constrained by other factors (climate and geology) that are of importance at a broader spatial and temporal scale. PMID- 15747411 TI - Cancer-fighting foods. Food choices and cancer risk. PMID- 15747412 TI - Health tips. Serving size matters. PMID- 15747413 TI - Many sinus headaches are actually migraines. PMID- 15747414 TI - Some people are resistant to aspirin's protective effects. PMID- 15747415 TI - Endometrial cancer. Better odds with early detection. PMID- 15747416 TI - Hand dermatitis. Treat your skin well. PMID- 15747417 TI - Dental surface repairs. Veneers and laminates. PMID- 15747418 TI - My wife and I spend the winter in the southern states. Recently, we saw a strange looking spider in our storage shed. Should we be concerned about being bitten? PMID- 15747419 TI - A friend recently told me about the new spray-on tanning that's being done in salons. Is it safe? PMID- 15747420 TI - Predicting phase noise in crystal oscillators. AB - In order to predict the phase noise in crystal oscillators an enhanced phase noise model has been built. With this model, the power spectral densities of phase fluctuations can be computed in different points of the oscillator loop. They are calculated from their correlation functions. The resonator-caused noise as well as the amplifier-caused noise are taken into account and distinguished. To validate this enhanced model, the behavior of a batch of 10 MHz quartz crystal oscillators is observed and analyzed. The tested batch has been chosen in a facility production. Their associated resonators have been selected according to the value of their resonant frequency and their motional resistance. Open-loop and closed-loop measurements are given. The phase noise of the overall oscillator working in closed loop is provided by the usual active method. Theoretical and experimental results are compared and discussed. PMID- 15747421 TI - Elimination of terrestrial rabies in Western European countries. AB - Since the late 1930s, the red fox (Vulpes vulpes) has been the main vector of rabies in Europe. Practically, decimation of fox population did not prevent the spread of the disease. The only efficient method to control wildlife rabies consisted in using oral vaccination by depositing vaccine baits containing a capsule or a plastic sachet filled with an attenuated anti-rabies liquid vaccine throughout fox habitats. Several live virus vaccines have been and are currently being used: the SAD B19 and SAD P5/88 are rabies strains attenuated in cell culture, the SAG1 and SAG2 strains are apathogenic mutants of an already attenuated rabies strain, and the VRG vaccine is an attenuated vaccinia virus recombinant coding for the rabies glycoprotein gene. These vaccines have different residual pathogenicity. SAG1 and SAG2 are pathogenic only for suckling mice inoculated by intracerebral and oral routes. VRG presents absolutely no rabies risk to humans and the environment and the residual pathogenicity of the vaccinia vector virus is very low even for humans. Other parameters such as the thermostability of the vaccine and the melting point of the bait casing are of utmost importance to guarantee the success of oral vaccination campaigns. Additionally, VRG is the only vaccine that did not interfere with maternal immunity in fox cubs, an important issue for spring campaigns. Successes and failure of national programmes confirm that whatever the ecological conditions, the same rules must be strictly followed to ensure the success of rabies elimination programmes: (i) considering the strategy, any rabies vaccination programme must be organised with the support of a national scientific team specially designated for the task that will have to apply the only methods that have already been proved successful elsewhere in Europe, including rabies surveillance, bait distribution calendar and pattern, and monitoring of this distribution; (ii) vaccination must be pursued for at least two years after the last reported case of rabies in the area; (iii) the choice of a low cost but poorly efficient and poorly stable vaccine does not prove to be cost-beneficial for successful elimination of rabies. Several European countries have become rabies-free: Belgium, Luxembourg, France, Italy, Switzerland, Finland and the Netherlands. Since the European Union is going comprise 25 countries from May 2004, all the scientific knowledge is available for establishing efficient and adapted oral programmes aimed at eliminating terrestrial rabies from this area. PMID- 15747422 TI - Health care reform brings new options. PMID- 15747423 TI - Formulary variability challenges physicians. PMID- 15747424 TI - Health care inflation takes a breather. PMID- 15747425 TI - OzFoodNet: enhancing foodborne disease surveillance across Australia: quarterly report, July to September 2004. PMID- 15747427 TI - Warning about indinavir and pregnancy. PMID- 15747426 TI - Surveillance of adverse events following immunisation for children aged less than 7 years, 1 January to 30 June 2004. PMID- 15747428 TI - Women testing tenofovir for HIV prevention. PMID- 15747429 TI - Atazanavir not to be used with proton pump inhibitors. PMID- 15747430 TI - Pill burden reduced for saquinavir. PMID- 15747431 TI - Collaboration begins on combined drug. PMID- 15747432 TI - Long-term care issue brief: end of life issues: year end report-2004. AB - End of life care is a broad issues that encompasses many issues. Three of the larger issues are assisted suicide, chronic pain management, and advance health care directives. Assisted suicide has emerged as one of the most controversial ethical issues in the medical community, and one that is complicated more by technological advances that have allowed people to live longer. But as policymakers debate assisted suicide policy, state lawmakers must also examine ways to improve pain management and end-of-life care for people who suffer from terminal illnesses. Lawmakers are also grappling with ways to make it easier for individuals to specify their end-of-life wishes through advance health care directives. PMID- 15747433 TI - Long-term care issue brief: family caregiving: year end report-2004. PMID- 15747434 TI - Long-term care issue brief: long-term care insurance: year end report-2004. PMID- 15747435 TI - Long-term care issue brief: nursing home liability insurance: year end report 2004. PMID- 15747437 TI - Medicaid and indigent care issue brief: Medicaid: benefits and services: year end report-2004. PMID- 15747436 TI - Long-term care issue brief: nursing home staffing standards: year end report 2004. PMID- 15747438 TI - Medicaid and indigent care issue brief: Medicaid: eligibility: year end report 2004. PMID- 15747439 TI - Medicaid and indigent care issue brief: Medicaid: provider reimbursement: year end report-2004. PMID- 15747440 TI - Medicaid and indigent care issue brief: Medicaid: waivers: year end report-2004. PMID- 15747441 TI - The constitutional implications of human cloning. PMID- 15747442 TI - Composition and temporal stability of gastrointestinal microbiota in irritable bowel syndrome--a longitudinal study in IBS and control subjects. AB - Irritable bowel syndrome (IBS) is a common intestinal disorder that includes continuous or recurrent intestinal pain and discomfort and altered bowel habits. The pathophysiology of IBS is incompletely understood, but it may involve an altered intestinal microbiota. The aim of the present study was to compare the composition and temporal stability of faecal microbiota of IBS patients and healthy controls by applying culture-based techniques and PCR-DGGE analysis. No difference in the prevalence or mean culturable manners of bacteroides, bifidobacteria, spore-forming bacteria, lactobacilli, enterococci or yeasts were observed between the IBS and the control groups, whereas slightly higher numbers of coliforms as well as an increased aerobe:anaerobe ratio was observed in the IBS group. PCR-DGGE revealed more temporal instability in the predominant bacterial population of IBS subjects than in controls. In 9 out of 21 IBS subjects and 5 out of 17 controls the PCR-DGGE profiles obtained from the samples of the same individual on different occasions (sampling points 0, 3 and 6 months) were clearly different. However, the instability in some of the IBS subjects could partly be explained by the antibiotic consumption during the study. The present study suggests that instability of intestinal microbiota may be involved in IBS. However, further studies are needed to associate the instability with specific IBS symptoms or with specific bacterial groups and species. PMID- 15747443 TI - Antibiotic resistance pattern of motile aeromonads from farm raised fresh water fish. AB - Motile aeromonads isolated from the intestines of farm-raised freshwater fish such as Catla catla, Labeo rohita and Ctenopharyngodon idella have been characterized to species level. Morphological and physiological grouping revealed 61% Aeromonas hydrophila, 30% Aeromonas caviae, 7% Aeromonas sobria and 2% which remained unidentified. Hemolytic activity was detected mostly in A. hydrophila, while only half of the A. sobria and A. caviae showed this activity. Antibiotic resistance patterns of the strains revealed that they had acquired a relatively higher resistance to oxytetracycline, amoxycillin, ampicillin, novobiocin and polymixin-B, implicating possible use of these antibiotics in the aquaculture systems. PMID- 15747446 TI - MAG's 2004 legislative voting record. PMID- 15747444 TI - Dynamics of storage reserve deposition during Brassica rapa L. pollen and seed development in microgravity. AB - Pollen and seeds share a developmental sequence characterized by intense metabolic activity during reserve deposition before drying to a cryptobiotic form. Neither pollen nor seed development has been well studied in the absence of gravity, despite the importance of these structures in supporting future long duration manned habitation away from Earth. Using immature seeds (3-15 d postpollination) of Brassica rapa L. cv. Astroplants produced on the STS-87 flight of the space shuttle Columbia, we compared the progress of storage reserve deposition in cotyledon cells during early stages of seed development. Brassica pollen development was studied in flowers produced on plants grown entirely in microgravity on the Mir space station and fixed while on orbit. Cytochemical localization of storage reserves showed differences in starch accumulation between spaceflight and ground control plants in interior layers of the developing seed coat as early as 9 d after pollination. At this age, the embryo is in the cotyledon elongation stage, and there are numerous starch grains in the cotyledon cells in both flight and ground control seeds. In the spaceflight seeds, starch was retained after this stage, while starch grains decreased in size in the ground control seeds. Large and well-developed protein bodies were observed in cotyledon cells of ground control seeds at 15 d postpollination, but their development was delayed in the seeds produced during spaceflight. Like the developing cotyledonary tissues, cells of the anther wall and filaments from the spaceflight plants contained numerous large starch grains, while these were rarely seen in the ground controls. The tapetum remained swollen and persisted to a later developmental stage in the spaceflight plants than in the ground controls, even though most pollen grains appeared normal. These developmental markers indicate that Brassica seeds and pollen produced in microgravity were physiologically younger than those produced in 1 g. We hypothesize that microgravity limits mixing of the gaseous microenvironments inside the closed tissues and that the resulting gas composition surrounding the seeds and pollen retards their development. PMID- 15747445 TI - Presentation of the 2004 ASP Distinguished Service Award to Austin J. MacInnis. PMID- 15747447 TI - Fludarabine: new indication. First-line treatment of CLL: unconvincing evidence. AB - (1) Oral chlorambucil is the reference first-line treatment for patients with symptomatic chronic lymphocytic leukaemia. Intravenous fludarabine is a second line option. (2) The indications for intravenous fludarabine have now been extended to cover first-line use. (3) The clinical evaluation dossier mainly contains data from two unblinded comparative trials. One trial compared intravenous fludarabine with chlorambucil in 509 patients for one year; the complete remission rate was significantly higher in the group receiving intravenous fludarabine (20% versus 4%) but, after 62 months of follow-up, the survival rate was no different (about 50% in both groups). Note that the dose of chlorambucil used in this trial may have been inadequate. (4) The other unblinded trial included 938 patients treated for six months with fludarabine, the CAP protocol, or the CHOP protocol. The median survival time did not differ among the three groups (67 to 70 months). (5) In the trial versus chlorambucil, the incidence of serious adverse events was significantly higher in the fludarabine group (55% of patients, versus 44%). The commonest serious adverse events were neutropenia, other haematological disturbances, and infections. (6) Oral chlorambucil is more convenient than intravenous fludarabine, which necessitates five infusions per month. Intravenous fludarabine costs 10 times more than oral chlorambucil. (7) In practice, chlorambucil remains the reference first-line treatment for chronic lymphocytic leukaemia. PMID- 15747448 TI - Teriparatide: new preparation. Osteoporosis: less well evaluated than alendronic acid. AB - (1) Oral alendronic acid is the reference drug for women with osteoporosis and a previous vertebral fracture. In a placebo-controlled trial in women who were also taking calcium and vitamin D, treatment with alendronic acid for three years reduced the incidence of symptomatic vertebral fractures (2.3% versus 5%) and wrist fractures (2.2% versus 4.1%) and, albeit with a lower level of evidence, the incidence of hip fractures (1.1% versus 2.2%). (2) Teriparatide, a biotech drug, reproduces the 34 N-terminal amino acids of parathormone. It is marketed in Europe for subcutaneous treatment of "proven" postmenopausal osteoporosis. (3) The cornerstone of the clinical evaluation dossier is a randomised placebo controlled double-blind trial in 1637 women also taking calcium and vitamin D. The two doses of teriparatide (20 micrograms/day and 40 micrograms/day), given for a median of 19 months, reduced the risk of new radiologically documented vertebral fractures (about 4% versus 14% in the placebo group) and spinal pain (about 16% versus 23% in the placebo group), but not the risk of hip fracture. (4) In a double-blind trial in 146 postmenopausal women also taking calcium and vitamin D, 40 micrograms/day teriparatide given subcutaneously for 14 months increased spinal mineral bone density significantly more than 10 mg/day alendronic acid given orally. The trial was not designed to show a difference in clinical outcome (fractures). (5) The main adverse effects of teriparatide reported to date are nausea, headache, cramp, hypercalcemia and hyperuricemia. (6) A rat study showed an increased risk of osteosarcoma. This tumour is rare in humans, and the number of patients so far enrolled in clinical trials is insufficient to document a possible increase in risk associated with teriparatide. (7) The need for daily subcutaneous injections and for refrigeration of the prefilled syringes are two notable disadvantages of teriparatide therapy. (8) In practice, alendronic acid is better assessed and remains the reference treatment, combined with calcium and vitamin D, for secondary prevention of osteoporotic fracture in postmenopausal women. PMID- 15747449 TI - Apropos correspondence. PMID- 15747451 TI - [Analysis of ECG on the staffs exposed to microwave in the radio calling signal station]. AB - 321 electrocardiographies (ECG) from the staffs exposed to microwave in a radio calling signal station were analysed. The subjects exposed to microwave were female computer operators, who worked over six months in the station. 107 female non-exposers were served as control. The intensities of microwave in work site were below the standard (20-50 mW/m2). The abnormal prevalence of ECG in the exposed group was 27.73%, which was significantly higher than that in the control group (11.21%, P<0.01). The prevalence of sinus arrhythmia in exposed group (19.63%) was significantly higher than that in the control group (5.60%, P<0.01). The P-R intervals were also lengthened in exposed group [(0.1554+/-0.021) ms] than that in control group [(0.1487+/-0.0227) ms, P<0.01]. No significant differences were found in the heart rate, QRS intervals, Q-T intervals, sinus bradycardia, sinus tachycardia and left axis deviation between exposed and control groups (P>0.05). It suggested that computer operators exposed to microwave of lower intensity in the radio calling signal station might lead to sinus arrhythmia, due to the disorder of vegetative nerve system. PMID- 15747450 TI - [The effects of sex and age on DNA adduct formation in human lung tissue]. AB - In order to evaluate the roles of sex and age in the formation of lung tissue DNA adducts, the levels of DNA adducts in human lung tissues from both patients with or without lung cancer were examined. The results showed there was a low correlation coefficient (r=0.11) between age and DNA adducts, but a significant difference in sex being observed. Smoking is a strong factor in the formation of DNA adducts. However, the role of endocrine can not be ruled out. PMID- 15747452 TI - [Evaluation study on determination of ZPP in blood with hematofluorometer]. AB - The authors studied the normalization and evaluation of the method detecting the zinc protoporphyrin (ZPP) by direct displaying blood fluorescent. The results revealed that this method was highly accurate and precise. The relative humidity not more than 90% had no influence to the results. The detection limit of this method was 0.1 microg/g. The linear range of standard curve was 0-15 microg/g, and the precision in batch RSD=0.27%-1.93%. On the accuracy side, the three concentration curving values of ZPP standard material in blood were all within the range of indefinition of given values. If the samples were not detected rapidly and the blood was diluted by heparin, it might be preserved at 4 degrees C for 3 days, at -8 degrees C for 8 days. The blank fluorescent value of glassy cover made in China is less than 3.5 microg/g, Hb can substitute KLI glassy cover. This method can analyse one sample per minute, has advantage of rapidity, convenience, accuracy and reliablity. PMID- 15747453 TI - [Study of the effects of pulse magnetic fields on hemorrheology of human body]. AB - The results show that middle shear specific viscosity and reductive viscosity (P<0.05), low shear specific viscosity and reductive viscosity of whole blood (P<0.01) decreased significantly in experimental group after 9-day of pulse magnetic fields, comparing with control group, high shear specific viscosity, low shear reductive viscosity (P<0.05) and middle and low shear specific viscosity (P<0.01) of whole blood in experimental group were lower after exposed than before exposed to pulse magnetic fields. The results suggest that the pulse magnetic fields may reduce specific and reductive viscosity of whole blood in human body. PMID- 15747454 TI - [Determinations of indole and methylindole in air by gas chromatography]. AB - Indole and methylindole in air were collected with alkaline multiport silicagel, desorbed with 1+1 acetone-CS2, and determined by GC-FID. Detectable limit is 3.5 mg/L. The minimum detectable concentration is 0.8 mg/m3. PMID- 15747455 TI - [Health assessment on safe treatment of night-soil on schistosomiasis endemic areas]. AB - The family size biogas tanks were built in the study site. Their effects were monitored. Faecal coliform and parasite eggs were examined before and after biogas digestion. The results of whole-year operation indicate that faecal coliform and parasite eggs can be reduced by 99.7% and 99.8%, respectively. The schistosome eggs survival experiment showed that the eggs all died off in 100 days of retention in biogas tanks. Therefore, the appropriate technology of nightsoil treatment can effectively interrupt the life cycle of schistosome. PMID- 15747456 TI - [Environmental pollution, human exposure and its health effect of sodium pentachlorophenate in schistosomiasis prevalent area]. AB - Sodium pentachlorophenate (Na-PCP) has been used in China for years as an molluscacide to kill oncomelania, which is an intermediate host of Schistosome. To evaluate the effects of its long-term successive usage on environment, human exposure and health, studies were carried out in Sichuan, Jiangxi, Jiangsu and Fujian provinces, with a time gap of more than one month between sample collection and last spray of Na-PCP. Results indicated that PCP contents in surface water, soil, sediment, animals and plants were significantly higher in studied areas than in control areas. The daily intake and the content in urine of PCP were also sigificantify higher in studied areas. But, there was no difference on physical and biochemical examinations except that a 22%-28% decrease of blood cholinesterase activity was found in studied areas. The health effect of impurities in Na-PCP, dioxins and furans, was assessed and discussed. PMID- 15747457 TI - [Analysis on current status of drinking water quality in rural areas of China]. AB - An investigation on drinking water quality in rural areas of 180 counties in 26 provinces, municipalities and autonomous regions of China was carried out. The population surveyed was 89.39 million. 69.6% of which was supplied with ground water. Central water supply systems served 47.1% of population. Quality of drinking water was graded according to the "Guidelines for Implementation of the 'Sanitary Standard for Drinking Water' in Rural Areas". The rate of population supplied with unqualified drinking water was 42.7%. The bacteriological indices of drinking water exceeded the standard seriously. Organic pollution occurred extensively. Some regions supplied with water of high concentration of fluoride. PMID- 15747458 TI - [Detection of hepatitis A virus in environmental samples by coupled reverse transcription and polymerase chain reaction]. AB - A direct reverse transcription coupled with polymerase chain reaction (RT-PCR) was used to detect HAV-seeded sea water, coastal sediment and Arca subcrenata, HAV strain treated by different kind of disinfection, and naturally occurring HAV in Arca subcrenata. The results showed that HAV could be detected in HAV-seeded sea water, coastal sediment and Arca subcrenata. HAV could also be detected in HAV strain disinfected by phenol, suggesting that inactive HAV could also show positive PCR. Only Arca subcrenata collected from Shanghai in 1988 showed positive result by RCR, but negative by immunofluorescence at the same time. This result demonstrated that HAV was not infectious owing to long time storage. The author's method has the advantage of involving fewer manipulations. PMID- 15747459 TI - [Dynamic study on blood lead levels of pregnant women and infants in a district of Beijing]. AB - Under strict quality control, the authors investigated the levels of PbB among 270 women from early pregnancy to delivery and their infants. The mean PbB levels of the first three months of pregnancy and delivery is 45.0 microg/L and 64.8 microg/L respectively. Analysis shows an increase of mean PbB level in the period of gestation, and a significant increase in the last three months. The mean PbB level of umbilical cord is 51.9 microg/L (10.4% umbilical cord PbB > 100 microg/L). The mean PbB level of infants increases with the months, it increases significantly after six months. Regression analysis confirmed the positive correlation between the PbB levels of first three and second three months of gestation, the PbB of delivery and umbilical cord, the PbB of umbilical coral and infants. The correlation coefficient of the PbB level of delivery and umbilical cord, the PbB level of infants at six months and at twelve months is 0.80 and 0.47 respectively. PMID- 15747460 TI - [The effect of smoking and passive smoking on thiocyanate content in saliva]. AB - The saliva thiocyanate contents were determined among smokers and passive smokers. The saliva thiocyanate contents of passive smokers in smoking environment and non-smoking environment were compared. The results showed that the saliva thiocyanate in smoking environment was higher than that in non-smoking environment (P<0.01). The results indicate that the saliva thiocyanate detecting is a direct and sensitive index for evaluating the hazard of passive smoking. PMID- 15747461 TI - [Bone mineral content and its influence factors of children aged 0-16 in a city and village]. AB - The radial and ulnar bone mineral content (BMC) was measured by single-photon absorptiometry, and the effects of multiple independent variables on BMC and bone width (BW) were explored using multiple regression analysis in 332 rural and city children aged 0-16 years in Henan province from 1994 to 1995. BMC, BW, BMC/BW and forearm length increased with age increasing, and the BMC in city children aged 0 6 years was higher than that in rural children. Radial BMC was positively related to BW in the measured sites. Age, body weight and height (or length) all had significantly positive and independent effects on radial BMC in both groups. The positive correlation of radial BMC with age was observed, however, the ratio of radial BMC to body weight was negative correlation. The BW in measured site and forearm length increased with ages. After adjusted for body size, there were no significant effects of sex, age and body weight on radial BW except height (length). This study shows that overall body size rather than adiposity is the important factor for bone BMC, so that it is needed to consider several variables, for example, BMC, BMD, BMC/body weight and BMC/age. PMID- 15747462 TI - [Effects of different form calcium on growth and tissue calcium level in rats]. AB - In order to solve the problem of calcium deficiency and to look for economic and efficient source of calcium, the effects of calcium carbonate, active calcium and calcium lactate on growth, development and tissue calcium level in rats were compared. Fifty-six 3-week old weaning rats were fed with calcium deficient diet (containing vitamin D 500 IU per kg diet) for 3 weeks, and then were divided into four groups randomly with 14 rats in each group, half male and half female. The diet of control group (A) was the basic diet, while the three experiment diets were supplemented with calcium carbonate (B), active calcium (C) and calcium lactate (D) (3000 mg calcium per kg diet), respectively. The experiment term was 12 weeks. The results showed that the body weight and length of calcium supplemented group were significantly higher than that of control group (P<0.05). Among the calcium supplemented groups, no significant differences were observed except the difference of body length between the group D and the group A in female. Calcium deficiency dramatically hindered the development with reduced dietary intake and decreased food consumption efficiency. The calcium levels in plasma, red blood cells and liver were significantly higher in the supplemented groups than that in the control group (P<0.05), however, there was no difference among the supplemented groups. No significant difference of calcium levels in muscle and heart was observed among all groups. Based on needs for reaching the RDA with additional 400 mg/d from present calcium status in Chinese population, the calcium carbonate is the most economic one and the ideal calcium source for supplementation. PMID- 15747463 TI - [Effect of nickel sulfate on the concentrations of T3, T4 and TSH in serum of rat]. AB - The effect of nickel sulfate on the concentrations of T3, T4 and TSH in sera of rats was studied. Forty wistar male rats were divided into four groups. The rats in groups 1, 2 and 3 were injected with 0.005 mol/L NiSO4, 0.01 mol/L NiSO4 and 0.02 mol/L NiSO4 [1 ml/(kg x d)] respectively, while the rats of group 4 were injected with normal saline, serving as control. Forty days later, the concentrations of T3 and T4 in sera of 0.01 mol/L NiSO4 and 0.02 mol/L NiSO4 groups were obviously decreased (P<0.05 or P<0.01). The concentrations of T4 in sera of 0.005 mol/L NiSO4 group was also decreased, compared with control group. There was no difference in the concentrations of T3 and T4 in sera among the groups with different doses of NiSO4. Between different dose groups and the control group, the concentrations of TSH in sera showed no significant difference (P>0.05). The proliferation of the epithelial cells of thyroid gland was noticed. The rough endoplasmic reticulam was exceedingly expanded. The nuclei were transformed. The results suggest that Ni may injure thyroid gland. PMID- 15747464 TI - [Method of cyanogen bromide-fluorimetry determination of trace amount of selenomethionine in grain and blood]. AB - Selenomethionine (SeMet) reacted with cyanogen bromide (BrCN) quantitatively forms CH3SeCN. After extracted with CHCl3, the Se of CH3SeCN is acid-digested to Se(IV). Then 2,3-diaminonaphthalene is used to determine the fluorescent Se value of 4,5-benzopiaselenol. The determination limit of this method was 3 ng/g SeMet. The accuracy of 10-500 ng Se in SeMet standard was 91.8%-97.6%. RSD was 1.9% 6.3%. Recoveries for grain and blood were 92.3%-96.7%. RSD was 2.7%-5.1%. The RSD for samples was 2.7%-9.0%. Selenocystine, selenocystiene selenite and methionine did not interfere with the determination. PMID- 15747465 TI - [A survey on mother's nutritional knowledge-attitude on the feeding of young child in Guangdong province]. AB - A survey on mother's nutrition knowledge-attitude on the feeding of young child was conducted in Guangdong province in 1995 among 414 mothers of 0 to 18 months old children. The results showed that the nutritional knowledge-attitude level of the mothers was low in both city and township. Only 51% of them had qualified level. The level of general nutritional concepts and the nutritional knowledge attitude during weaning period were significantly lower than that of breastfeeding and follow-weaning. It suggests that the promotion of nutritional education in the feeding of young children is urgent in Guangdong province especially in township and rural area. It is necessary to focus on the improvement of nutritional knoweledge in weaning period and general nutrition concepts. PMID- 15747466 TI - [Characterization of volatile organic compounds from indoor materials]. AB - Volatile organic compounds emitted from eight indoor materials including paint, wallpaper glue, maticolor coat, floorwax floor covering and air freshener and were analyzed by headspace GC-FID and GC-MS. About 3-30 organic compounds were detected from each material. It was shown that the emitted compounds and emission rate from different materials were different. The emitted compounds included paraffin, olefine, alcohol, aldehyde, ether, ester and aromatic compounds. The results of dynamic experiments in test chamber showed that the chamber concentration of volatile organic compounds from paint vs. time data follows a double exponent equation. PMID- 15747467 TI - [Experimental research on the prediction of human body thermal responses in special hot environment]. AB - The assessment of thermal environment is very important in the research of human body-thermal environment system. On the basis of systematic study, the characteristics and the components of human body-thermal environment system have been analyzed, the stationary mathematical model of human body-thermal environment system was established. Prediction and Assessment System for Thermal Environment (PASTE system) was developed. It consists of thermal environment monitor and simulator based on the IBM PC/XT computer with suitable hardware and software. Thermal environment monitor can measure fifteen parameters continuously including dry bulb temperature, dewpoint temperature, air velocity and thermal radiation in six directions. The simulator can simulate different heat exchange processes of human body-thermal environment system and human thermal responses. PASTE system is integrated with monitor and computer simulator. Test results showed that the differences between experimental result and predicted result were about +/-10%. PMID- 15747468 TI - [Neurospychiatric symptoms in HIV infected patients and the role of efavirenz]. AB - Efavirenz has now become commonly used to treat HIV infection. Neuropsychiatric disorders have been reported in patients treated with efavirenz. Several factors often make it hard to determine the cause of these disorders: HIV infected patients take many different drugs, they may suffer from various organ diseases, and may also be heavily affected by problems in their everyday life. The French experts group working on neuropsychiatric side effects of efavirenz has undertaken a review of these disorders with the aim to identify: (1) semiology, (2) epidemiology in the global population, in HIV infected patients, and in patients treated with efavirenz. The expert group suggests recommendations to manage these disorders. PMID- 15747469 TI - [Activity of nalidixic acid and fluoroquinolones on Escherichia coli strains isolated from non-complicated urinary infections (reseau TSN - France, 1999 - 2001)]. AB - OBJECTIVE: The authors studied the susceptibility of 1,647 non-repeat isolates of Escherichia coli to quinolones and fluoroquinolones. METHOD: The strains were isolated from non-complicated urinary infections in women 18-64 years of age. Data was provided by the TSN Database France, a real time electronic database which collects antibiotic susceptibility results and patient demographic data. The data was collected from 1999 to 2001 in 63 French hospital laboratories, each using their own routine test methods. Quantitative data was interpreted (S, I, R) according to CA-SFM breakpoint guidelines. RESULTS: Ninety-eight and 94,6 % of the strains were susceptible to ciprofloxacin and nalidixic acid respectively. Cross resistance was assessed as well as intrinsic difference in activity within the fluoroquinolone class. Current fluoroquinolones are still highly efficient, and ciprofloxacin is the most active. CONCLUSION: Since 1996, little change in resistance to fluoroquinolones has been observed. These results confirm the choice of fluoroquinolones as first intention therapy as recommended by consensus conferences. PMID- 15747470 TI - [Efficacy of cefotiam hexetil in acute maxillary sinusitis, with a short five day vs ten day treatment]. AB - OBJECTIVE: The aim of this multicentric, randomized, double blind study with direct individual benefit, was to compare two therapeutic regimens of cefotiam hexetil administration, 5 days vs 10 days, in acute maxillary sinusitis. METHOD: The study was conducted in ambulatory patients treated by general practitioners according to AFSAPS (French agency for sanitary safety) guidelines for treatment of acute maxillary sinusitis. Five hundred and fifty three GPs included 1042 patients presenting with acute maxillary sinusitis in the study from December 2000 to July 2001. Patients were randomly treated with cefotiam hexetil 200 mg bid over a 5 day period followed by 5 days of placebo, or with cefotiam hexetil 200 mg bid over a 10 day period. RESULTS: No significant difference was noted in each treatment group. Radiography performed in 72.2% of included patients confirmed the diagnosis in 78.8% of the cases. No significant difference occurred in the number and percentage of cured patients. In the ITT analysis (1018 patients) the clinical cure rates were respectively 85.5% and 85.3% in the 5 day and in the 10 day treatment groups, In the PP analysis (800 patients) the clinical cure rates were respectively 88.6% in each group. The low incidence of adverse effects (3.36%) was confirmed in both groups. CONCLUSION: A 5 day course of cefotiam hexetil 200 mg bid is as effective as a 10 day course in the treatment of acute maxillary sinusitis in adults. PMID- 15747471 TI - [Regional variations of adult population vaccinal status]. AB - OBJECTIVE: The authors had for aim to study the regional variations of vaccination coverage in adult population. METHODS: Cross-sectional epidemiological study, including a representative sample of 2122 general practitioners and analyzed according to eight French regions. RESULTS: Six thousand two hundred sixty-nine patients, 44 +/- 15 old, were included in the study. 90.5% of the patients were vaccinated against diphtheria, 94.4% against tetanus, and 92.6% against poliomyelitis. No difference between regions was noted but the rate of patients with up to date vaccination strongly differed, ranging from 54.8% (Paris/Ile de France) to 64.3% (South West) for diphtheria, from 60.4% (Paris/Ile de France) to 73% (South West) for tetanus, and from 58.8% (Paris/Ile de France) to 69.8% (South West) for poliomyelitis. Vaccination coverage against measles, parotiditis, hepatitis A and B, typhoid fever, yellow fever also significantly differed between regions and the only ones evenly distributed were vaccination against tuberculosis, meningococcus, and pneumococcus. CONCLUSION: These results show insufficient vaccination coverage against diphtheria, tetanus, and poliomyelitis in all regions and show other important variations in vaccination coverage between regions, especially for hepatitis A and B. PMID- 15747472 TI - Surveillance of influenza in Apulia, Italy, 1999-2000, 2000-2001, 2001-2002, and 2002-2003 seasons. AB - BACKGROUND: The objective of this study was to evaluate, within the Italian National Influenza Epidemiological and Virological Surveillance, the rate of vaccination coverage, the incidence of Influenza Like-Illness (ILI), the incidence of Acute Respiratory Illness (ARI), and to identify the virus strains circulating in Apulia from 1999 to 2003. METHODS: Vaccination coverage rates were calculated based on the number of doses administered to individuals > 65 years of age. Every week, sentinel physicians reported ILI and ARI cases having occurred among their patients. Voluntary general practitioners (GPs) and paediatricians (Ps) collected oropharyngeal swab samples from patients suspected with ILI. Influenza viruses were isolated and identified by cell culture (MDCK cells) and RT-PCR. Virological surveillance was carried out by the ISS, in collaboration with a network of peripheral laboratories. RESULTS: In Apulia, vaccination coverage progressively increased to 68.6% during the 2002-2003 season. The analysis of ILI cases showed higher incidence rates during the 1999-2000 and 2002 2003 seasons. ARI rates appeared to have a more constant trend. ILI and ARI incidence rates were higher in the 0-14 year age group. CONCLUSION: The increase in vaccination coverage rates and implementation of the network of clinical, and epidemiological and virological surveillance are fundamental for the control and prevention of influenza. PMID- 15747473 TI - [Resources and organization of 124 health-care institutions in South-East France for standard precautions and isolation]. AB - OBJECTIVES: The authors had aimed to evaluate resources and organization necessary for applying guidelines issued 5 years earlier in various institutions: standard precautions, septic isolation, prevention against spread of multidrug resistant bacteria. MATERIAL AND METHODS: Volunteer institutions were surveyed for hygiene product consumption, architectural requirements, inventory of protocols, description septic patients'management, and available personnel. RESULTS: One hundred and twenty-four institutions (40,784 beds) were included in the study. Eleven percent had no hygiene physician or nurse; the rates of personnel specialized in hygiene practice were 0.4 physicians per 800 beds and 0.8 registered nurses per 400 beds. Eighty-eight percent of the studied institutions had a protocol for standard precautionary measures, 77% had a septic isolation protocol. A multidrug-resistant bacteria identification sheet was attached to examination reports in 87% of cases. Multidrug-resistant bacteria screening was practiced by 18.1% of the institutions. Hygiene product consumption for 1000 days of hospitalization was 7861 disposable gloves, 2.3 1 of hydroalcoholic solution, and 63 disposable gowns. 28.9% of the wards lacked water hand washing points, 32.2% had no sinks, and 48.5% had no local equipment maintenance. In addition, 40.7% of the beds were in single rooms, 4.8% of the institutions had no single rooms. Differences were observed depending on specialties and institutions. CONCLUSION: Analysis of consumption shows insufficient application of standard precautions, notably for hydroalcoholic solutions. The number of single rooms is acceptable, architectural requirements were not adequate in too many wards. These results can explain some problems encountered in applying the guidelines. PMID- 15747474 TI - [Crusted scabies in a patient with chronic GVH: difficulty of diagnosis and consequences]. PMID- 15747476 TI - [Seroprevalence of hepatitis E virus in Casablanca]. PMID- 15747475 TI - Tuberculosis of the parotid gland: case report and literature review. PMID- 15747477 TI - 'Tis the season--of high grassroots legislative activity. PMID- 15747478 TI - A defense of ethical relativism. PMID- 15747479 TI - Justice in medicine and public health. PMID- 15747480 TI - Medical custom and medical ethics: rethinking the standard of care. PMID- 15747481 TI - Epiphanic knowledge and medicine. PMID- 15747482 TI - Genetic testing of children for late onset disease. PMID- 15747483 TI - Is evaluating ethics consultation on the basis of cost a good idea? PMID- 15747484 TI - Prenatal testing, reproductive autonomy, and disability interests. PMID- 15747485 TI - Respect for equality and the treatment of the elderly: declarations of human rights and age-based rationing. PMID- 15747486 TI - The age-indifference principle and equality. PMID- 15747487 TI - Toleration and healthcare ethics. PMID- 15747488 TI - Conscription of cadaveric organs for transplantation: a stimulating idea whose time has not yet come. PMID- 15747489 TI - Presumed consent: an international comparison and possibilities for change in the United States. PMID- 15747490 TI - Fever: facts, fiction, physiology. PMID- 15747491 TI - Having fun with professionalism. PMID- 15747492 TI - How would you use graduate education? PMID- 15747493 TI - Consider this when choosing a graduate program. PMID- 15747494 TI - Coaching others for a higher level of performance. PMID- 15747495 TI - Manage conflict creatively. PMID- 15747496 TI - Doctoral degrees have options. PMID- 15747497 TI - Successfully integrating e-learning with critical care nursing education. PMID- 15747498 TI - AACN new subspecialty certification exams. PMID- 15747499 TI - Human metabolism and metabolic interactions of deployment-related chemicals. AB - It has been suggested that chemicals and, more specifically, chemical interactions, are involved as causative agents in deployment-related illnesses. Unfortunately, this hypothesis has proven difficult to test, because toxicological investigations of deployment-related chemicals are usually carried out on surrogate animals and are difficult to extrapolate to humans. Other parts of the problem, such as the definition of variation within human populations and the development of methods for designating groups or individuals at significantly greater risk, cannot be carried out on surrogate animals, and the data must be derived from humans. The relatively recent availability of human cell.fractions, such as microsomes, cytosol, etc., human cells such as primary hepatocytes, recombinant human enzymes, and their isoforms and polymorphic variants has enabled a significant start to be made in developing the human data needed. These initial studies have examined the human metabolism by cytochrome P450, other phase I enzymes, and their isoforms and, in some cases, their polymorphic variants of compounds such as chlorpyrifos, carbaryl, DEET, permethrin, and pyridostigmine bromide, and, to a lesser extent, other chemicals from the same chemical and use classes, including solvents, jet fuel components, and sulfur mustard metabolites. A number of interactions at the metabolic level have been described both with respect to other xenobiotics and to endogenous metabolites. Probably the most dramatic have been seen in the ability of chlorpyrifos to inhibit not only the metabolism of other xenobiotics such as carbaryl and DEET but also to inhibit the metabolism of steroid hormones. PMID- 15747500 TI - Drug bioactivation, covalent binding to target proteins and toxicity relevance. AB - A number of therapeutic drugs with different structures and mechanisms of action have been reported to undergo metabolic activation by Phase I or Phase II drug metabolizing enzymes. The bioactivation gives rise to reactive metabolites/intermediates, which readily confer covalent binding to various target proteins by nucleophilic substitution and/or Schiff's base mechanism. These drugs include analgesics (e.g., acetaminophen), antibacterial agents (e.g., sulfonamides and macrolide antibiotics), anticancer drugs (e.g., irinotecan), antiepileptic drugs (e.g., carbamazepine), anti-HIV agents (e.g., ritonavir), antipsychotics (e.g., clozapine), cardiovascular drugs (e.g., procainamide and hydralazine), immunosupressants (e.g., cyclosporine A), inhalational anesthetics (e.g., halothane), nonsteroidal anti-inflammatory drugs (NSAIDSs) (e.g., diclofenac), and steroids and their receptor modulators (e.g., estrogens and tamoxifen). Some herbal and dietary constituents are also bioactivated to reactive metabolites capable of binding covalently and inactivating cytochrome P450s (CYPs). A number of important target proteins of drugs have been identified by mass spectrometric techniques and proteomic approaches. The covalent binding and formation of drug-protein adducts are generally considered to be related to drug toxicity, and selective protein covalent binding by drug metabolites may lead to selective organ toxicity. However, the mechanisms involved in the protein adduct-induced toxicity are largely undefined, although it has been suggested that drug-protein adducts may cause toxicity either through impairing physiological functions of the modified proteins or through immune-mediated mechanisms. In addition, mechanism-based inhibition of CYPs may result in toxic drug-drug interactions. The clinical consequences of drug bioactivation and covalent binding to proteins are unpredictable, depending on many factors that are associated with the administered drugs and patients. Further studies using proteomic and genomic approaches with high throughput capacity are needed to identify the protein targets of reactive drug metabolites, and to elucidate the structure-activity relationships of drug's covalent binding to proteins and their clinical outcomes. PMID- 15747501 TI - Enantioselective pharmacokinetics of ibuprofen and involved mechanisms. AB - Although dexibuprofen (S-ibuprofen) was marketed in Austria and Switzerland, the racemate at various formulations is still extensively used worldwide, and there are no indications that the racemate will be replaced by the single enantiomer. Thus, elucidation of the characteristics and involved mechanisms of the chiral pharmacokinetics of racemic ibuprofen is of special importance for the understanding of the pharmacological and toxicological consequences, and for prediction of the clinically potential drug interactions and influence of the pathological states. Stereoselective pharmacokinetics and metabolism are common features for chiral nonsteroidal antiinflammatory drugs (NSAIDs) and especially for 2-arylpropionic acid derivatives characterized with a chiral center adjacent to the carboxyl group. Although the enantioselective pharmacokinetic characteristics of different NSAIDs should be treated case by case, they share similar mechanisms underlying the protein binding, metabolism and chiral inversion. Ibuprofen was the most extensively researched drug in terms of chiral characteristics and mechanisms. Therefore, elucidation of the mechanisms derived from research on ibuprofen may provide better understanding and prediction of other chiral drugs. This article attempts to elucidate the chiral pharmacokinetics and involved mechanisms of ibuprofen in comparison with other NSAIDs based on recent developments. Topics on history of ibuprofen, enantioselective analysis method, absorption, protein binding, conventional metabolism, metabolic chiral inversion, gene polymorphism, and biochemical developments were included. It is worth mentioning that some underlying biochemical mechanisms, especially for the metabolic chiral inversion and ethnic differences still remain to be seen. Further research is required to develop human-resourced researching model and to provide more evidence concerning the site of inversion, species variation, CYP450 gene polymorphisms, and biochemical mechanisms. PMID- 15747502 TI - Autoantibodies against CYP2D6 and other drug-metabolizing enzymes in autoimmune hepatitis type 2. AB - Autoimmune hepatitis (AIH) is a disease of unknown etiology, characterized by liver-related autoantibodies. Autoimmune hepatitis is subdivided into two major types: AIH type 1 is characterized by the detection of ANA, SMA, ANCA, anti-ASGP R, and anti-SLA/LP. Autoimmune hepatitis type 2 is characterized to be mainly related with drug-metabolizing enzymes as autoantigens, such as anti-LKM (liver kidney microsomal antigen)-1 against CYP2D6, anti-LKM-2 against CYP2C9-tienilic acid, anti-LKM-3 against UGT1A, and anti-LC1 (liver cytosol antigen)-1 and anti APS (autoimmune polyglandular syndrome type-1) against CYP1A2, CYP2A6, and others. Anti-LKM-1 sera inhibited CYP2D6 activity in vitro but did not inhibit cellular drug metabolism in vivo. CYP2D6 is the major target autoantigen of LKM-1 and expressed on plasma membrane (PM) of hepatocytes, suggesting a pathogenic role for anti-LKM-1 in liver injury as a trigger. Anti-CYP1A2 was observed in dihydralazine-induced hepatitis, and radiolabeled CYP1A2 disappeared from the PM with a half-life of less than 30 min, whereas microsomal CYP1A2 was stably radiolabeled for several hours. Main antigenic epitopes on CYP2D6 are aa 193-212, aa 257-269, and aa 321-351; and D263 is essential. The third epitope is located on the surface of the protein CYP2D6 and displays a hydrophobic patch that is situated between an aromatic residue (W316) and histidine (H326). Some drugs such as anticonvulsants (phenobarbital, phenytoin, and carbamazepine) and halothane are suggested to induce hepatitis with anti-CYP3A and anti-CYP2E1, respectively. Autoantibodies against CYP11A1, CYP17, and/or CYP21 involved in the synthesis of steroid hormones are also detected in patients with adrenal failure, gonadal failure, and/or Addison disease. PMID- 15747503 TI - Cellular export of drugs and signaling molecules by the ATP-binding cassette transporters MRP4 (ABCC4) and MRP5 (ABCC5). AB - Like other members of the multidrug resistance protein (MRP)/ABCC subfamily of ATP-binding cassette transporters, MRP4 (ABCC4) and MRP5 (ABCC5) are organic anion transporters. They have, however, the outstanding ability to transport nucleotides and nucleotide analogs. In vitro experiments using drug-selected or transfected cells indicated that these transport proteins, when overexpressed, can lower the intracellular concentration of nucleoside/nucleotide analogs, such as the antiviral compounds PMEA (9-(2-phosphonylmethoxyethyl)adenine) or ganciclovir, and of anticancer nucleobase analogs, such as 6-mercaptopurine, after their conversion into the respective nucleotides. This may lead to an impaired ability of these compounds to inhibit virus replication or cell proliferation. It remains to be tested whether antiviral or anticancer chemotherapy based on nucleobase, nucleoside, or nucleotide precursors can be modulated by inhibition of MRP4 and MRP5. MRP4 also seems to be able to mediate the transport of conjugated steroids, prostaglandins, and glutathione. Furthermore, cyclic nucleotides (cyclic adenosine monophosphate and cyclic guanine monophosphate) are exported from cells by MRP4 and MRP5. This may modulate the intracellular concentration of these important mediators, besides the action of phosphodiesterases, as well as provide extracellular nucleotides for a possible paracrine action. In this line, tissue distribution and subcellular localization of MRP4 and MRP5 specifically in smooth muscle cells (MRP5), platelet-dense granules (MRP4), and nervous cells (MRP4 and MRP5), besides the capillary endothelium, point not only to a possible function of these transporters as exporters in cellular defense, but also to a physiological function in signaling processes. PMID- 15747504 TI - [The effects of anesthetics on G-protein-coupled receptors]. AB - Although anesthetics have been often used clinically, the mechanisms of action of anesthetics have not yet been clarified. Recently, major advances have been made in our understanding of the physiology and pharmacology of G-protein-coupled receptor (GPCR)-mediated signaling. Several lines of studies have shown that GPCRs are targets for anesthetics and that some anesthetics inhibit the functions of Gq-coupled receptors, including muscarinic acetylcholine (ACh) M1, metabotropic type 5 glutamate, 5-hydroxytryptamine (5-HT) type 2 A, and substance P receptors. Many additional GPCRs have been classified as "orphan" receptors (oGPCRs) because their endogenous ligands have not been identified yet. Given that known GPCRs are targets for anesthetics, these oGPCRs may represent a rich group of receptor targets for anesthetics. This review highlights the effects of anesthetics on Gq-coupled receptors, and discusses whether GPCRs other than Gq coupled receptors, and proteins that convey GPCR signals are also targets for anesthetics. PMID- 15747505 TI - [Effects of simultaneous epidural administration of ropivacaine and morphine on the post-operative pain in the gynecologic patients]. AB - BACKGROUND: We examined the effects of simultaneous epidural administration of ropivacaine with morphine on the level of the post-operative METHODS: Forty-one patients were assigned to one of three groups [ropivacaine (R), ropivacaine + morphine (RM) or morphine (M)]. In the R group, 5 ml of 1% ropivacaine bolus was administered just before the skin incision followed by infusion of 0.2% ropivacaine (5 ml x hr(-1)) during the first 48 hours after the operation. In the RM and M groups, 5 ml of 1% ropivacaine + 2 mg of morphine bolus was administered just before the skin incision followed by infusion of 0.2% ropivacaine (RM group, 5 ml x hr(-1)) or saline (M group, 5 ml x hr(-1)) + 4 mg x day(-1) of morphine during the first 48 hours after the operation. RESULTS: The score of post operative pain in the R group is higher than that of the MR group or that of M group. There is no difference between the score of post-operative pain of the MR group and that of the M group. CONCLUSIONS: These data suggested that simultaneous epidural administration of ropivacaine with morphine produces no beneficial effect as compared with morphine alone. PMID- 15747506 TI - [Anesthetic management by total intravenous anesthesia with propofol, pentazocine and ketamine]. AB - BACKGROUND: The authors evaluated the efficacy of anesthetic management by total intravenous anesthesia with propofol, pentazocine and ketamine. METHODS: Thirty five patients for mastectomy were anesthetized by propofol, pentazocine and ketamine. Patients were divided into two groups by age; one is patients under 61 years of age and the others are patients above 61 years. Analysis was made retrospectively. Anesthesia was induced with propofol and ketamine and was maintained with propofol infusion and intermittent administration of vecuronium with 40% oxygen in air. Pentazocine was administrated as a bolus dose before incision. RESULTS: There were no differences in the patient background except age and height between the two groups. After induction of anesthesia, systolic and diastolic blood pressures decreased compared with those before induction in both groups. Systolic and diastolic blood pressures and heart rate increased after tracheal intubation, but the hemodynamics remained stable after the start of surgery. The induction and maintenance doses of propofol were not different between the two groups. Patients above 61 years had smaller dosage of pentazocine compared with those in patients under 61 years. The dosage of ketamine was not different between two groups. Awakening time in about 80% of patients was within 15 minutes and is not different between the two groups. Postoperative pain relief was good in both groups. Incidence of nausea and vomiting was 25% and was not the different between the two groups. CONCLUSIONS: Total intravenous anesthesia with propofol, pentazocine and ketamine would be useful to stabilize hemodynamic state, to obtain rapid recovery and to provide effective postoperative pain relief. PMID- 15747507 TI - [Changes in core temperature associated with lower extremity tourniquet--the influence of ambient temperature and warming equipments]. AB - BACKGROUND: We investigated changes in core temperature associated with lower extremity tourniquet (TQ) under two different ambient temperatures (1) and two different warming equipments (2) under general anesthesia combined with lumbar epidural anesthesia. METHODS: (1) The values of core temperature at ambient temperature of either 22 degrees C (n = 15) or 20 degrees C (n=15) were recorded after induction of anesthesia, at start of TQ application, at the termination of TQ application, and 14 minute after TQ release. (2) The values of core temperature using either air-forced warming or active heated i.v. at ambient temperature 20 degrees C were recorded at four points as mentioned above. RESULTS: (1) Changes in core temperature were not observed during TQ application at ambient temperature both 20 degrees C and 22 degrees C. Core temperatures in both groups decreased significantly after TQ release, and core temperatures at termination of TQ application and after TQ release at ambient temperature 20 degrees C were significantly lower than those at ambient temperature 22 degrees C. (2) Significant increases in core temperatures using two different warming equipments were observed at termination of TQ application and after TQ release at ambient temperature 20 degrees C. Core temperatures using air-forced warming were maintained during the investigation, though significant decrease in core temperature using active heated i.v. was recorded after TQ release. CONCLUSIONS: Air-forced warming maintains core temperature efficiently associated with lower extremity tourniquet. PMID- 15747508 TI - [Cardiovascular effects of human atrial natriuretic peptide during renal transplantation]. AB - BACKGROUND: We evaluated the cardiovascular effects of human atrial natriuretic peptide (hANP) in the recipients of renal transplantation. METHODS: Anesthesia was maintained by inhalation of nitrous-oxide and isoflurane in oxygen, with epidural block. The recipients were divided into three groups; one group received no hANP infusion as control and the other groups received continuous infusion of hANP at the rate of either 0.05 microg x kg(-1) x min(-1) or 0.1 microg x kg(-1) x min(-1). Intravenous infusion of hANP was started at the anastomosis of renal artery after the fresh frozen plasma was loaded to achieve PCWP over 17 mmHg. In each group, we examined cardiovascular changes by using a pulmonary artery catheter and transesophageal echocardiography. The measurements were done before and after 15 minutes of hANP infusion. RESULTS: In comparison with control, the decreases in PCWP and CVP were significant in the 0.1 microg x kg(-1) x min(-1) group. An increase in CI and the reduction of CVP were significant in 0.05 microg x kg(-1) x min(-1) group, when compared with control group. In the 0.1 microg x kg(-1) x min(-1) group, the reductions of PCWP and CVP and MAP were significant, but the significant increase in CI was characteristic in the 0.05 microg x kg(-1) x min(-1) group. CONCLUSIONS: We conclude that the low-dose infusion of hANP in the recipients of renal transplantation is useful for the optimal anesthetic care because of the cardiovascular improvement. PMID- 15747510 TI - [Percutaneous tracheostomy to the patient with coagulopathy]. AB - BACKGROUND: Generally, coagulopathy is a relative contraindication for surgical tracheotomy. METHODS: Fourteen patients with severe respiratory failure and coagulopathy underwent elective percutaneous tracheostomy using Portex percutaneous tracheostomy kit. RESULTS: Apart from minor bleeding episodes during and after the procedures which were controlled promptly, no other complications occurred. CONCLUSIONS: If careful precautions are observed, we believe that percutaneous tracheostomy is safe even in patients with severe respiratory failure and coagulopathy. PMID- 15747509 TI - [Anesthetic management of patients with interstitial cystitis during intravesical resiniferatoxin therapy]. AB - The management of patient with interstitial cystitis (IC) remains a challenge because no single agent has proven effective. IC is a chronic sterile inflammatory disease of the bladder of unknown etiology characterized by urinary frequency, urgenecy, nocturia and lower abdominal pain. We experienced anesthetic management of five patients with IC during intravesical resiniferatoxin (RTX) therapy. RTX is associated with irritative urinary symptom during bladder instillation. The patients with IC had bladder instillation with 100 ml of 10(-8) M RTX solution for 30 min. The first patient received combined spinal-epidural anesthesia (CSEA), and the others general anesthesia. The patient with regional anesthesia had no critical troubles related to circulatory status during the procedure, but increases of blood pressure after instillation of RTX were observed in two patients receiving general anesthesia. In spite of the increase in blood pressure during general anesthesia, regional anesthesia should not be used, because the effect of RTX on the spinal cord has to be maintained. PMID- 15747511 TI - [Thoracic epidural analgesia is effective in perioperative pain relief for uterine artery embolization]. AB - BACKGROUND: Uterine artery embolization (UAE) has become widely employed in Japan. Although several methods of anesthesia and analgesia are performed for UAE, pain control does not appear to be satisfactory. We report a series of UAE, successfully managed using thoracic epidural analgesia. METHODS: Before UAE an epidural analgesic catheter was inserted at T 10-11. Local anesthetic and morphine sulfate were administered through the catheter. After the UAE patients received patient-controlled epidural analgesia (PCEA) for 24 hours. The next day, patients were treated with diclofenac sodium 25 mg suppository every 8 hours. RESULTS: In all cases, early pain was controlled well by epidural analgesia. Late pain was controlled by combining epidural analgesia with diclofenac sodium. Nausea and pruritus due to morphine sulfate occurred in some cases, but disappeared after discontinuation of PCEA. CONCLUSIONS: Several phases of severe pain are seen perioperatively in UAE. Because thoracic epidural analgesia is easily administered and the dosage of the drugs used effectively controlled, it is a practical method for perioperative pain control for UAE. PMID- 15747512 TI - [Anesthesia for cesarean section in a patient with cor triatriatum]. AB - Cor triatriatum is a rare congenital cardiac anomaly, in which left ventricular filling is impeded by obstructive membrane in the left atrium. We administered spinal anesthesia for cesarean section in a patient with cor triatriatum (type III A1) with congestive heart failure. We optimized hemodynamics with the aid of pulmonary artery cathter. In general, cor triatriatum involves similar hemodynamic profiles to mitral stenosis and thus tachycardia should be avoided during anesthesia. However, in our patient, increasing the heart rate to 80-90 beats x min(-1) was beneficial in maintaining adequate systemic blood pressure and cardiac output. Spinal anesthesia could be a method of choice for cesarean section in a patient with cor triatriatum when adequate hemodynamic monitoring is available. PMID- 15747513 TI - [Hypoxemia after gastrointestinal endoscopy under general anesthesia in a patient with unilateral diaphragmatic paralysis]. AB - A 59-year-old woman with suspected hypopharyngeal cancer was scheduled for biopsy using direct laryngoscopy. Her preoperative chest X-ray showed asymptomatic left unilateral diaphragmatic paralysis. Anesthesia was induced with propofol and maintained with oxygen, nitrous oxide and sevoflurane. The region was difficult to reach during direct laryngoscopy. Therefore, gastrointestinal endoscopy was performed. Air was injected through the scope to aid viewing. After tracheal extubation, oxygen saturation as indicated by pulse oximetry (SpO2) decreased to 91% from 97% although oxygen was delivered via a mask at 6 l x min(-1). A chest radiograph showed that the air-filled gastrointestinal tract had elevated the left dome of the diaphragm. After the stomach was suctioned via a nasogastric tube, respiration was assisted via a mask with continuous positive airway pressure. Then, SpO2 returned to 99%. An air-filled dilated stomach may increase the risk of respiratory dysfunction in patients with unilateral diaphragmatic paralysis. PMID- 15747514 TI - [Severe airway obstruction relieved by sedation using sevoflurane in a pediatric patient with tracheobronchomalacia]. AB - A 3-month-old baby with trisomy 18 syndrome was scheduled for tracheostomy under general anesthesia because of the prolonged tracheal intubation. Immediately after transferring the patient to the operating table, the patient suddenly began crying and coughing, resulting in severe hypoxia. The patient's lungs could not be ventilated by manual and positive pressure ventilation, and airway obstruction could not be relieved until the respiratory effort spontaneously decreased. We started to administer sevoflurane on the recommendation of pediatricians who had successfully treated the patient with sedation using either midazolam or trichlorethylphosphate in similar situations. After sevoflurane administration, the sedated patient never developed the respiratory effort, and the lungs could be ventilated by manual and positive pressure ventilation without difficulty. The patient was diagnosed as tracheobronchomalacia as a result of intraoperative flexible bronchoscopy performed through tracheostomy tube, revealing significant narrowing of both the trachea and mainstem bronchus lumens. Sedation using sevoflurane may be helpful in maintaining airway patency in the pediatric patient with tracheobronchomalacia. PMID- 15747516 TI - [Successful management of a man with fulminant myocarditis using percutaneous cardiopulmonary support]. AB - Fulminant myocarditis is a fetal disease characterized by a distinct viral prodrome, sudden onset of severe hemodynamic compromise, and marked myocardial inflammation. One possible therapy to improve the poor prognosis of such patients may be the implantation of circulatory support systems that allow myocardial recovery. We report here successful management of a patient with fulminant myocarditis using percutaneous cardiopulmonary support (PCPS), intra-aortic balloon pump (IABP), and continuous hemodiafiltration (CHDF). A 37-year-old Japanese man suddenly experienced cardiopulmonary dysfunction shortly after general fatigue, and was diagnosed as having fulminant myocarditis. PCPS was immediately initiated because catecholamine infusion and IABP were not enough to support circulation. Although severe dyskinesis was observed on his admission, cardiac function recovered twelve days after PCPS initiation with ejection fraction from 16% to 73%. Renal and hepatic failure also recovered with the improvement of cardiac function. We describe our clinical experiences in cardiogenic shock after acute fulminant myocarditis and discuss therapeutic guidelines for the use of PCPS, with its management and complications. PMID- 15747515 TI - [Anesthetic management for an infant with tetralogy of fallot with absent pulmonary valve]. AB - Tetralogy of Fallot with absent pulmonary valve (TOF/APV) is a rare disease, and characterized by massive dilatation of pulmonary artery. Bronchial compression by the aneurysm-like dilated pulmonary artery sometimes results in severe respiratory compromise. Surgery including closure of VSD and right ventricular outflow construction is often needed in infancy. But, even after the surgical correction, patients often suffer from right ventricular failure due to right ventriculotomy and poor development of pulmonary capillaries. Therefore, preventing pulmonary hypertension is important during the perioperative period. We report an infant with TOF/APV. This 3 month-old boy developed respiratory distress suddenly. After the induction of anesthesia with sevoflurane, fentanyl, and vecuronium, continuous infusion of alprostadil and olprinone was started. Cardiopulmonary bypass (CPB) was instituted smoothly. VSD was closed and right ventricular outflow tract was reconstructed. Dilated pulmonary arteries were resected and plicated. Weaning from CPB was easy and postoperative course was uneventful with mild pulmonary hypertension. In the cases predisposed to right ventricular failure like TOF/APV, olprinone may be useful. PMID- 15747517 TI - [Emergency microlaryngosurgery for a huge post-intubation laryngeal granuloma]. AB - A 55-year-old woman who had undergone esophagectomy 3 months earlier developed a huge post-intubation laryngeal granuloma. She was scheduled for emergency microlaryngosurgery. Anesthesia was induced with 8% sevoflurane in oxygen under spontaneous respiration. After confirming no airway compromise, the spontaneous respiration was obtunded by forced manual ventilation without using a muscle relaxant. The trachea was intubated easily with an endotracheal tube. Anesthesia was maintained with sevoflurane and nitrous oxide supplemented with intravenous fentanyl. The granuloma removed was 11 x 8 mm in diameter. There were no respiratory complications during and after anesthesia and surgery. PMID- 15747519 TI - [A case report of a patient who developed hemiparaplegia with multiple cerebral infarction during thoracoabdominal aortic aneurysm repair]. AB - To protect the spinal cord during thoracoabdominal aortic aneurysm repair, motor evoked potentials (MEP) monitoring and cerebrospinal fluid drainage are often employed. Herein, we report a case, where intraoperative diminishment of motor evoked potentials was accompanied by multiple cerebral infarction. A 63-year-old man underwent elective surgery for both thoracoabdominal aortic aneurysm and abdominal aortic aneurysm. He had a past history of cerebral infarction, resulting in Wernicke aphasia but no paralysis. Preoperative magnetic resonance angiography and echocardiography revealed occlusion of the intercostal and lumbar arteries, mild aortic regurgitation, and atherosclerotic lesions at the aortic arch as well as descending aorta. Anesthesia and muscular relaxation were maintained with fentanyl, propofol, and continuous administration of vecuronium at 0.5 mg x kg(-1) x h(-1). The thoracoabdominal aortic aneurysm was repaired under distal aortic perfusion with femorofemoral bypass. After terminating the bypass, we found that the MEP at the lower limb had disappeared. Although we reconstructed intercostal arteries under mild hypothermia and partial bypass, the amplitude of MEP remained very low. Suspecting spinal cord ischemia, we performed cerebrospinal fluid drainage immediately after the operation. On the postoperative day 4, when we stopped the cerebrospinal fluid drainage and propofol administration, his level of consciousness was poor and brain CT revealed multiple cerebral infarction. On the postoperative day 30, he was discharged from an intensive care unit with complications of hemiplagia and paraplegia. Although cerebrospinal fluid drainage may be recommended to protect spinal cord during thoracoabdominal aortic aneurysm repair, we should consider performing brain CT to exclude a risk of brain herniation secondary to cerebrospinal fluid drainage if there is a possibility of cerebral incidents. PMID- 15747518 TI - [A case of continuous infusion of milrinone during continuous venovenous hemofiltration causing high plasma concentration of milrinone]. AB - A 79-year-old woman was undergoing chronic hemodyalysis. She received a continuous infusion of milrinone of 0.25 microg x kg(-1) x min(-1) for 77 hours, 0.2 microg x kg(-1) x min(-1) for 39 hours and 0.1 microg x kg(-1) x min(-1) for one hour due to heart failure after mitral valve repair. While receiving continuous infusion of milrinone, she was undergoing continuous venovenous hemofiltration. After infusion of milrinone for three days no arrhythmia was seen except atrial fibliration, but blood pressure remained low. After stopping infusion of milrinone, the plasma concentration of milrinone was 792.7 ng x ml( 1) (therapeutic plasma concentration: 100-200 ng x ml(-1)). Now few data are available regarding the pharmacokinetic of milrinone in patients with severe renal failure and during CVVH. Therefore we should design a dose modification for intravenous milrinone in renal failure patients. PMID- 15747520 TI - [Regional discrepancies of numbers of anesthesiologists and certified training hospitals in Japan]. AB - BACKGROUND: We reported previously the existence of regional discrepancies in the numbers of anesthesiologists to the general population, the numbers of all medical doctors and hospital beds in Japan. In the present study, we investigated the regional discrepancies of the numbers of anesthesiologists and hospitals with department of anesthesiology from medical facility's point of view in Japan. METHODS: We compared the numbers of regular members, Board Certified Anesthesiologists (BCAs) and Certified Training Hospitals (CTHs) of the Japan Society of Anesthesiologists (JSA) to the general hospitals among forty-seven prefectures and nine Districts of JSA. Moreover, we compared the numbers of the general hospitals and CTHs to the general population. RESULTS: There were significant correlation coefficients between the number of the general hospitals and the number of JSA regular members or BCAs or CTHs, and the general population and the number of the general hospitals or CTHs in all prefectures. The mean number of JSA regular members per one general hospital was 0.96 persons and the maximum difference among all prefectures (MDAP) was 4.0 times. Tokyo District of JSA had the largest numbers (1.96 persons) of JSA regular members per one general hospital and the maximum difference among all Districts (MDAD) was 2.8 times. The mean number of BCAs per one general hospital was 0.54 persons and the MDAP was 4.2 times. Tokyo District of JSA had the largest numbers (1.01 persons) of BCAs per one general hospital and the MDAD was 2.6 times. The percentage of CTHs to all general hospitals was 9.1% and the MDAP was 5.4 times. The MDAD was 1.9 times. The mean number of the general hospitals per one hundred thousands population was 7.25 facilities and the MDAP was 4.3 times. Kyushu District of JSA had the largest numbers (11.50 facilities) but Tokai District of JSA had the smallest numbers (5.24 facilities) of the general hospitals per one hundred thousands population and the MDAD was 2.1 times. The mean number of CTHs per one hundred thousands population was 0.66 facilities and the MDAP was 3.3 times. Hokkaido District of JSA had the largest numbers (0.93 facilities) and Chugoku Shikoku District of JSA had the second largest numbers (0.87 facilities) but Tokai District of JSA had the smallest numbers (0.45 facilities) of CTHs per one hundred thousands population and the MDAD was 2.1 times. CONCLUSIONS: There are remarkable regional discrepancies in the numbers of anesthesiologists and CTHs to the general hospitals and in the number of CTHs to the general population in Japan. We suspect that the regional discrepancies in the field of anesthesiology would influence the quality and form of corresponding clinical practice. PMID- 15747521 TI - [Prevalence and risk factors for latex sensitization among anesthesiologists]. AB - BACKGROUND: Although powdered latex surgical gloves are predominantly used in Japanese healthcare facilities, the prevalence of latex sensitization among anesthesiologists has not been investigated. METHODS: The authors surveyed 60 anesthesiologists of 16 facilities with questionnaires and measurements of specific IgE antibodies against latex and other aeroallergens by AlaSTAT microplate immunoassay. Sensitization was defined as positive if the specific IgE concentration was not less than 0.70 IU x ml(-1). RESULTS: With surgical gloves 63.3% of anesthesiologists used powdered latex gloves, compared to 10.2% with examination gloves. The prevalence of latex sensitization was 33.3%, with a peak of 70% (7/10) in ages 45-49. In multivariate analysis, anesthesiologists of the facilities where more than 50% of them used powdered latex surgical gloves had a 6.0-fold risk of latex sensitization (95% CI 1.7-21.5; P=0.006). Histories of atopic dermatitis, asthma, or food allergy were also considered as risk factors (OR 3.8; 95% CI 1.1-13.7; P=0.038). The history of allergic rhinoconjunctivitis was not associated with latex allergen, but with Japanese cedar pollen. No relation was observed between latex and timothy pollen. CONCLUSIONS: Our results suggest that surgical powdered latex gloves were the major predisposing factor for latex sensitization measured by latex-specific IgE among anesthesiologists. PMID- 15747522 TI - [New study on the history of anesthesiology--(11) three Japanese doctors who observed William TG Morton's ether anesthesia at the Gross Clinic in 1860]. AB - In January of 1860 the Tokugawa Shogunate dispatched a group of Japanese mission headed by Masaoki Niimi, the first ambassador, to Washington D.C. to exchange ratification of Japan-United States treaty of commerce and amity. On May 22nd the ratification was approved. They left Washington D.C. to move to Baltimore and Philadelphia to visit various sites and institutions. The Niagara sailed from New York with the Japanese mission on board on June 30th. In Philadelphia three Japanese medical doctors visited Jefferson Medical College to observe an operation of lithotomy by Dr Samuel D Gross, professor of surgery. Ether was given by Dr William TG Morton, the man that had succeeded in the public demonstration of ether anesthesia at Massachusetts General Hospital in Boston on October 16, 1846. The doctors might have brought new information on ether anesthesia to Japan, but they only left few traces of providing the information to medical professionals in Japan. PMID- 15747523 TI - ["Oral healthiness score for 8020" predicts loss of teeth in village residents]. AB - OBJECTIVE: It is important for people to maintain an appropriate lifestyle through out life to enjoy a healthy life. We have already developed a health check questionnaire, "8020 Oral Healthiness Score", in an endeavor to help people aged 80 keep more than 20 teeth. The health check consists of ten questions chosen from our previous residents' study. Named the "Sawayaka Score" it has been in use since 1999. In the present study, we focused on the results of a three year follow-up to determine whether the Score may predict tooth loss for screening purposes. METHODS: A total of 716 village residents who participated in medical and dental checkups in 1999 as the baseline year were followed in T village of Aichi-Prefecture. The total numbers of teeth lost were examined after 1-, 2- and 3-years and odds ratios with 95% confidence intervals (CI) were generated for analysis of screening. RESULTS: At the 1999 baseline, the average number of retained teeth was 23.7 +/- 6.2 (mean +/- standard error) (23.0 +/- 6.8 in males, and 24.4 +/- 5.5 in females), and the average oral health score (mean +/- standard error) was 13.1 +/- 3.9 points (12.8 +/- 4.0 in males, 13.4 +/- 3.9 in females). A low score of 4-8 at the baseline predicted major tooth loss, while an average score was associated with loss of only one or 2 teeth loss in the residents. The present study confirms that persons with a favorable lifestyle tend to retain more teeth. CONCLUSION: It can be concluded that the 8020 Oral Healthiness Score is a useful tool to support residents' oral health promotion and predict tooth loss. PMID- 15747524 TI - [Relationship between the white blood cell count and development of electrocardiographic ST-T abnormalities. Insights from annual health examinations of urban residents]. AB - OBJECTIVE: Inflammation has been shown to play a role in atherosclerosis and coronary heart disease. This study was designed to examine the relationship between the baseline white blood cell (WBC) count and development of electrocardiographic ST-T abnormalities. METHODS: The results of annual health examinations conducted in the city of A, Osaka Prefecture, from 1985 to 1997 were evaluated. At the initial examination, carried out during the period 1985-1988, 1,213 women and 201 men, who were current non-smokers without hypertension, were free from ST-T abnormalities. We focused on cases with new ST-T abnormalities identified during the 12-year period. WBC counts were categorized as low (< 50th percentile) or high (> or = 50th percentile), and were divided into quintiles. Cox proportional hazards models were used to test for correlations. RESULTS: The age-adjusted mean WBC count in both men and women were higher for cases with new ST-T abnormalities than for those with normal ECG findings. For men, the percentage of positive cases with a high WBC count was significantly greater. Multivariate models showed that the relative risk (RR) of new ST-T abnormalities for cases with a high WBC count as compared with those with a low WBC count was 7.16 (P<0.001) for men and 1.50 (P< 0.001) for women. The quintiles for men showed a step-wise increment in the rate per 1,000 person-years but no such tendency was observed for women. The higher the quintile in men, the higher the RR was, and the RR in the highest quintile was approximately ten times that in the lowest quintile. For women, a similar trend was observed, but the association between the RR and the WBC count was weaker. CONCLUSIONS: These results confirm that the WBC count is significantly associated with development of ST-T abnormalities, and that an elevated WBC count is a marker for an increased risk of ST-T abnormalities. In women, this relationship appears less prevalent than in men. PMID- 15747525 TI - [Critical thinking on "home visits" through the case method using the teaching material, "the twelfth child birth"]. AB - OBJECTIVE: To use case method seminars in order to critically assess "home visits" for public health nursing practice. METHOD: "The twelfth childbirth" was developed for use as material for the case method in 2002. This case story involves the dilemma of a public health nurse as to whether or not she should intervene in the "private affairs" of a married couple. Case teaching was performed in two seminars during the period from July 2002 to February 2003. Participants in the first seminar were 18 health professionals (13 public health nurses and five supervisors) in Japan, and nine health professionals (8 midwives and one physician) from Cambodia took part in the second. RESULTS: For the problems from the case, the participants in the two seminars made their decisions analytically. Decision making was informative and took different directions. The public health nurses in Japan advocated a collaborative intervention with other health professionals. On the other hand, the midwives from Cambodia selected an approach involving heavy commitments to family planning undertaken by individual midwives. From the discussion in the seminars, this case story was satisfied through the following: (1) the participants used the information in the case to address the problem; (2) the participants thought analytically in order to evaluate potential solutions; and (3) the participants had sufficient information for analysis in the case. CONCLUSION: Both seminars provided good opportunities to enhance critical thinking on "home visits" as a tool for intervention and to develop thinking skills needed for public health nursing practice. PMID- 15747526 TI - [Factor structure of the SOC scale 13-item version in Japanese university students]. AB - The purpose of the present study was to examine the construct validity of Antonovsky's (1987) sense of coherence scale (SOC) 13-item version, with a sample of Japanese university students. Subjects were 1,110 university students who completed a self-administered questionnaire including items on individual characteristics (age, sex), the Zung self-rating depression scale (SDS) and the SOC scale. Exploratory factor analysis of the SOC scale yielded an interpretable two-factor solution with "comprehensibility-manageability" and "meaningfulness". Confirmatory factor analysis demonstrated an a priori defined SOC second-order factor model composed of comprehensibility-manageability and meaningfulness factor to have an acceptable fit (chi-square=327.065, df=64, GFI=0.957, CFI=0.872, RMSEA= 0.061). Moreover, structural equation modeling showed that there are theoretically consistent relationships among individual SOC factors and depression. In particular, the meaningfulness factor proved better predictor of depression than the comprehensibility-manageability factor. These findings support the construct validity of the SOC scale and suggest that two SOC sub scores may provide clinically applicable information, rather than the total score. PMID- 15747527 TI - [A questionnaire survey of mental health and welfare in city health centers prior to the partial amendment of the mental health and welfare law going into effect]. AB - OBJECTIVE: The situation of mental health of health centers in cities prior to the partial amendment of the Mental Health and Welfare Law going into effect was investigated to provide data relevant to mental health and welfare in cities. METHOD: 81 health centers were recruited from designated, core, and ordinance designated cities as well as the 23 sections of Tokyo City, and were supplied into questionnaires by mail. RESULTS: Less than 10 percent of the health centers had played a central role in establishing rehabilitation facilities. Regarding active support, small community-based workshops were most commonly subsidized. Only 13.3% of the health centers in Tokyo City provided support for daily life training facilities, while 60% had rehabilitation facilities. Seventy percent provided in-home services. More specifically, in contrast to the 64.7% of health centers in the designated cities which provided home-helper services, the figure was only 10.0% in ordinance-designated cities. Regarding the 2000 social adaptation training program, this was put into effect by less than half of the health centers in Tokyo City. No patients were admitted to rehabilitation facilities through 28.6% of the health centers, or underwent the consultation for rehabilitation facilities at 37.1%. Participation in care manager training sessions was low in Tokyo City but high in the other areas. Applicants for the health and welfare handbook and medical expense assistance for people with mental disorders were interviewed in 40.0% of Tokyo City's health centers and in over 70% of the others. There were problems with counter application in 50% of health centers in the core cities but not many in the remainder. More than 70% of health centers in the designated cities and Tokyo put the transfer system based on Article 34 of the law into effect but the percentages were lower for core and ordinance-designated cities. Patients were transferred at 34.2% of the health centers where the transfer system was enacted. Most of the designated cities and Tokyo established a council for mental health and welfare. However, this was the case for only 21.4% of the core cities and many of the ordinance-designated cities did not even consider this matter. Many health centers answered that welfare policy had advanced with the partial amendment. CONCLUSION: The study revealed major gaps among health centers and specific problems with different background types. PMID- 15747529 TI - [A survey on education of epidemiology and biostatistics in nursing programs at Japanese universities]. PMID- 15747528 TI - [Depressive symptomatology and some relevant factors. Investigations at a junior college in Hokkaido, Japan]. AB - PURPOSES: Recently in Japan, major depression, a depressive state, and suicide are increasingly becoming social problems requiring preventive intervention. However, only few investigations have been performed of actual conditions of mental health. The present study was therefore planned and carried out to examine relations between stress and relevant factors in adolescents and to search for clues for preventive intervention. METHODS: A total of 184 students in a junior college in Hokkaido were the subjects of this cross-sectional survey. In October 2003, age, sex, health-related factors, social supports, stressors, loci of control, coping styles, and depressive status were investigated. After excluding data from incomplete responses, we ultimately analyzed 153 students (mean age = 19.9, 141 females). RESULTS: Regardless of the CES-D categorization, after conducting univariate logistic regression analysis, we found that cognitive style and consciousness of stress were significantly associated with depressive symptomatology. The association remained significant even after multivariate logistic regression analysis. CONCLUSIONS: Cognitive style may influence depression in adolescents. We now need to consider preventive interventions applying the theory of cognitive-behavioral therapy. PMID- 15747530 TI - [New radiologic technique: advancement of multislice row of data CT for abdominal imaging]. PMID- 15747531 TI - [Barrett epithelium and Barrett carcinoma from a pathological viewpoint]. PMID- 15747532 TI - [Strategy for diagnosis and medical management of Barrett's esophagus]. PMID- 15747533 TI - [Barrett's epithelium and Barrett's adenocarcinoma from the viewpoint of surgery]. PMID- 15747534 TI - [Evaluation of 13C-urea breath test to confirm eradication of Helicobacter pylori]. AB - This study aimed to evaluate the effectiveness of the 13C-urea breath test (UBT) for assessment of Helicobacter pylori eradication after treatment. One hundred twenty six patients were enrolled with 85 receiving proton pomp inhibitor based triple therapy. They were underwent upper gastrointestinal endoscopy with biopsies for diagnosis and assessment of H. pylori infection using culture, histology, rapid urease test (RUT) and 13C-UBT. Assessment of eradication needs to be performed 4 weeks or more after completion of treatment. Breath samples were taken 15 minutes after the ingestion of 100 mg 13C-urea. Breath samples were analyzed on a mass spectrometer system. The gold standard for H. pylori infection was a positive culture or positive histology + positive RUT; negative for infection was defined as negative results of all three biopsy tests. Based on ROC curves, the most appropriate cut-off value for diagnosis of H. pylori infection was identified as 2.5/1000, which provided 96.2% sensitivity, 100% specificity, and 96.8% accuracy as judged by the gold standard. However, when confirming the eradication of H. pylori, it was 3.5/1000, which provides for 100%, 95.8%, and 96.5%, respectively. Ten patients (11.8%) had delta13C values that were 2.5 5.0/1000 4-12 weeks after therapy. Eight patients were considered cured of H. pylori infection, and 2 were considered to still have H. pylori infection following 13C-UBT, serology, and H. pylori specific antigen test. The false positive rate of 13C-UBT was 9.4% (8/85). When the grey zone of 13C-UBT was set at a level of 2.5 to 5.0/1000 (2.5 > : negative, 5.0 < or = : positive) after eradication therapy, the sensitivity and specificity of 13C-UBT was 100% and 98.4% compared to the gold standard. It was concluded that to avoid false positive results of 13C-UBT, the grey zone of 13C-UBT needs to be set at a level of 2.5 to 5.0/1000; thus improving the accuracy of test for the assessment of eradication of H. pylori infection. PMID- 15747535 TI - [A case of acute portal and mesenteric venous thrombosis treated with urokinase via superior mesenteric artery]. PMID- 15747536 TI - [A case report of advanced gastric remnant cancer treated with extended resection accompanied with reconstructions of the portal vein and the hepatic artery with resultant disease free survival for one year and eight months]. PMID- 15747537 TI - [A case of hepatocellular carcinoma with bone metastasis treated by TS-1 and CDDP]. PMID- 15747538 TI - [A case of pancreatic mucinous cystadenoma with a high serum CA19-9 level]. PMID- 15747539 TI - [Skin rash enhanced by ampicillin in a patient with infectious mononucleosis]. PMID- 15747541 TI - [Reliability of students' evaluations of university teaching: an analysis of four facet data by a generalized model and structural equation modeling]. AB - In our study, we examined the reliability of students' evaluation of university teaching. First, we analyzed four-facet data (teacher x rater x viewpoint x order) by the generalizability theory approach, and estimated the variance components for the facets as a G study. Then, we evaluated the reliability from the point of view of the generalizability coefficient (a reliability-like coefficient that is used when a decision concerns the relative ordering of individuals) and the index of dependability (an index that is used when a decision focuses on the absolute level of an individual's performance independent of others' performance), and examined how those indices change as the number of raters and viewpoints are changed. With these analyses we found conditions needed to maintain sufficient reliability of the evaluation for various situations. We also introduced a new method to evaluate the reliability at each level of facet by using structural equation modeling. By this method, one can examine the reliability of students' evaluation of university teaching more specifically. PMID- 15747540 TI - [Age and gender differences as factors related to depressive symptoms among community-dwelling middle-aged and elderly people]. AB - This study was conducted to identify factors associated with depressive symptoms among males and females in two age groups; the middle-aged (40-59 years) and elderly (60-79 years). Subjects were 2211 community-dwelling people (1115 males and 1096 females). Depressive symptoms were estimated by Center for Epidemiologic Studies Depression Scale (CES-D). Marital status, self-rated health, activities of daily living, age awareness, locus of control, and social support were assessed using self-administered questionnaires and interviews. In all groups, poor subjective health and external locus of control showed significant positive association with depressive symptoms. Age awareness was associated with depressive symptoms in female groups. Impairment of instrumental activities of daily living and lower social support were associated with depressive symptoms in the elderly groups. The findings suggested that there might be age and gender differences in the impact of factors associated with depressive symptoms. PMID- 15747543 TI - [Study of the inhibitory mechanism in retrieval-induced forgetting with the explicit/implicit memory paradigm]. AB - Anderson, Bjork, and Bjork (1994) devised a retrieval-practice paradigm, thereby showing that remembering can cause forgetting, termed retrieval-induced forgetting. The present research examined what aspect of memory was suppressed by the inhibitory mechanism accompanied by remembering, by using an implicit memory task. Thirty-six undergraduates learned a list of 36 category-instance pairs. They performed directed retrieval practice on some critical items from the learned list by completing a series of cued-fragment recall. Following a distractor task, they then performed either an explicit or an implicit memory task probed by category-plus-first letter pairs. The results indicated the typical pattern of retrieval-induced forgetting in the explicit memory task, but not in the implicit memory task. This suggests that the inhibitory mechanism affects the explicit retrieval process (accessibility) rather than representation of items in memory (availability). PMID- 15747542 TI - [Study of paranoid ideation in college students: an approach using the diathesis stress model]. AB - A recent study reported that normal people who have social anxiety and attention to social and interpersonal cues show paranoid ideation (Martin & Penn, 2001). The purpose of the present study was to examine whether people with these diatheses show paranoid ideation in the diathesis-stress framework. Questionnaires were administered to 177 college students three times. At Time I, social anxiety and attention to social and interpersonal cues were assessed. These variables were used in Martin and Penn (2001) as the diathesis factor of paranoid ideation. At Time 2, paranoid ideation during the period, and at Time 3, paranoid ideation and stress events between Time 2 and Time 3 were assessed. Setwise hierarchical multiple regression analysis was used to predict the change in the incidence of paranoid ideation from Time 2 to Time 3. Result revealed significant interaction effects between social anxiety and stress. Students with high score on the social anxiety scale showed an increase in paranoid ideation when they experienced more stress events. PMID- 15747544 TI - [Effects of mere subliminal exposure on trait judgments and the role of stereotyped knowledge]. AB - This study investigated the effects of repeated exposures to male and female targets on trait impressions and the role of stereotyped knowledge for the target's social category in impression formation process. The participants were repeatedly exposed to slides of male and female faces for subliminal durations. For each of 12 pairs containing both previously presented slide and newly presented slide, the participants made forced-choice liking judgments (Experiment 1), trait judgments (Experiment 2) and recognition judgments (Experiments 1 and 2). It was found that participants' attitude toward the targets became more positive, even though target recognition was not significantly greater than the chance level. Yet, when the dimension of judgment was stereotypically associated with the target's social category, exposure effects were obtained for the targets whose social category and its dimension were inferentially matched, but not obtained for the targets whose social category and its dimension were not inferentially matched. Some theoretical implications of the role of social category information in the mere exposure phenomenon are discussed. PMID- 15747545 TI - [Concordance of facial reactions to facial expressions]. AB - Participants watching a facial expression of emotion tend to respond with the same facial expression. This facial concordance is well known for happiness, but not for other emotions. The present study investigated whether facial expressions of basic six emotions induce facial concordance in participants by average-face method. Facial reactions of 20 subjects were videotaped while watching a facial expression of six emotions performed by amateur actors in a computer display. The six average faces were made from corresponding facial expression of emotions in the display. Newly chosen 62 subjects were asked to classify those average faces into six categories of emotion. The facial concordance was found for happiness and surprise, but not for disgust and fear. However, for average face of anger and that of sadness, classifications were divided into a few categories. This result suggests a possibility that an average faces might have included ambiguous or different faces. It may be necessary to conduct re-classification not with the average-face but with individual faces. PMID- 15747546 TI - [Influences of recollective experience and self-other differences in episodic retrieval on a following recall test]. AB - This study investigated influences of recollective experience and self-other differences in episodic retrieval on a following recall test. In the learning phase, subjects were assigned to either self-episodic or other-episodic retrieval condition. In both conditions, subjects were asked to retrieve an episode associated with each trait word, and reported "Remember", "Know" or "No". Immediately after the phase, they performed an arithmetic problem for five minutes, and then were given the surprise recall task. "Remember"-reported words were better recalled than "Know"-reported words. And there was no difference in recall between self-episodic and other-episodic retrieval conditions. These results suggest that an existence of recollective experience in episodic retrieval during encoding, produces better recall performance. PMID- 15747547 TI - [Developing the Japanese version of the Adult Attachment Style Scale (ECR)]. AB - This study attempted to adapt into Japanese the Adult Attachment Style Scale (ECR: Experiences in Close Relationships inventory) that was constructed by Brennan, Clark, and Shaver (1998), based on 14 existing scales. Of 387 respondents, 231 who reported having been or are currently involved in romantic relationships were employed for final analysis. We examined validities of the Japanese version of ECR in the two ways: (1) Examining the correlations between "Anxiety" and Self-esteem scale by Rosenberg (1965) which were theoretically related to Self-view, and the correlations between "Avoidance" and Other-view scale by Kato (1999b) which were theoretically related to Other-view; (2) whether or not ECR represents the features of four attachment styles as classified by Relationship Questionnaire (RQ; Bartholomew & Horowitz, 1991). The results supported our expectations. This Japanese version of ECR was demonstrated to have adequate psychometric properties in validity and reliability. PMID- 15747548 TI - [Review of the gender research in cross-cultural psychology since 1990: conceptual definitions and methodology]. AB - A review of the cross-cultural research on gender in psychology since 1990 reveals (1) conceptual confusion of the definitions of sex, gender, man, and woman; (2) diversification, refinement, reification, and a problem-solving orientation in the research topics; and (3) the possibility of the elucidation of the psychological sex-difference mechanism in relation to the biological sex differences. A comparison of 1990 and 2000 cross-cultural psychological articles published in "Sex Roles" found that overall, the research is Western-centered and some methodological problems remain to be solved concerning the measures and the sampling. These findings lead to the following suggestions for cross-cultural research on gender to resolve the problems and contribute to the development of psychology in general: (1) use of an operational definition for conceptual equivalence; (2) conducting more etic-approach research; (3) avoiding ethnocentric or androcentric research attitudes; (4) use of a theoretical framework; (5) strict examination of methodologies; and (6) examination of the specific context of participants in terms of cultural diversity, dynamics of husband-wife relationships, and relationships with husbands and fathers. PMID- 15747549 TI - [Paired comparison analysis by using structural equation modeling: Scheffe's method and its improvements]. AB - The purpose of this study is to propose a way to express and implement paired comparison analysis in a framework of structural equation modeling (SEM). By this method, one can perform paired comparison using widely available SEM programs and can develop a variety of models for specific purposes. Here, three models are shown. One is a model for performing basic paired comparison by using SEM. Another is an expanded model which makes it possible to apply analysis of variance (ANOVA) or regression analysis to the result of paired comparison. A third model is for paired comparison of latent factors. All models are illustrated with actual numerical examples. PMID- 15747550 TI - [Trust and cooperation: a comparison of in-group preference and trust behavior between American and Japanese students]. AB - American and Japanese students, 44 and 38, respectively, participated in an experiment, and played a game together in seven- or eight-person groups. The game was a repeated version of bilateral trust game: Prisoner's Dilemma (PD) with choice of dependence. In no-information condition, participants were not told that some of the group members were from another country. In information condition, they were told that about half of the members were Japanese and the rest were Americans. We examined whether or not people trusted ingroup members (those from the same country) more than outgroup members, and whether or not they cooperated with ingroup members more than outgroup members. We found no evidence of ingroup bias in terms of trust and cooperation, and we did not find significant differences in the levels of trust or cooperation between those found among Americans and those among Japanese. On the other hand, it was found that American participants were more sensitive than Japanese counterparts, to information regarding the past trust behavior of other players when they were deciding whom they trust and whether or not they reciprocate another's trust. PMID- 15747551 TI - [Status quo bias in conflict situations]. AB - The present study examined status quo bias in hypothetical conflict situations. Based on Lewin's (1935) typology, four situations were described: approach, avoidant, ambivalent, and no information conflicts. In Study 1 in which 15 male and 68 female undergraduates participated, status quo bias was found in their responses to no information as well as avoidant conflict situations. In Study 2, 29 male and 99 female undergraduates performed decision tasks, and status quo bias was found in ambivalent as well as avoidant conflict situations. In addition, in Study 3, with 25 male and 75 female undergraduates, status quo bias was found in no information, avoidant, and ambivalent conflict situations. In general, strong status quo bias was found in no information as well as avoidant conflict conditions, whereas it was consistently absent in approach conflict situation. These results suggest that status quo bias is a strategy to avoid decisions. PMID- 15747552 TI - [Effects of initial familiarity and source recollection on confidence ratings of recognition memory]. AB - Recent studies have shown the importance of metamemory functions such as confidence ratings in memory performance. However, the question of how people evaluate the accuracy of their own memory remains unresolved. Contrary to the previous studies that indicated the correlation between accuracy and confidence, we examined the influence of subjective source recollection on confidence levels of recognition judgments through two experiments. Participants were first requested to learn two word-lists presented in male and female voices respectively, and to make the yes-no recognition and source judgments--"male", "female" or "don't know". The results showed that participants had higher confidence on recognition judgments when they made source attribution than when they chose "don't know" (Experiment 1), and that participants found the recognition items more familiar when they later made source attribution irrespective of its correctness (Experiment 2). These findings suggest that the high initial familiarity to the recognition items induces the subjective experience of recollection, and that such recollective experience leads to high confidence on recognition memory regardless of the accuracy. PMID- 15747553 TI - [Assessment of the distance between categories in rating scales by using the item response model]. AB - This study aimed to assess the distance between adjacent categories of rating scales. It is common practice to treat ordinal variables as interval-scaled variables in the analysis of rating scales. Strictly speaking, however, ordinal scale data should be treated as such, since there is little reason and assurance that they are equivalent to interval variables. In view of this practice, this study proposes a method to assess the interval of rating scales, and analyzes empirical data in order to examine the results obtained by the method. This method is based upon the generalized partial credit model which is one of item response theory (IRT) models. The experiment was carried out on two data sets that differed only on the verbal phrasing of the rating. Main results of the study were: 1) the difference in item content (positive or negative) affects the width of a neutral category; and 2) the distance between categories differs significantly reflecting the difference in verbal phrasing. PMID- 15747554 TI - [Study of structured selection interviews at a Japanese corporation]. AB - In the job hiring process where long-term employment is expected, it is considered vital for the employer to form a mutually trusting relationship with applicants throughout the selection process. In this study, we attempted to develop a structured interview for new graduate hiring at a Japanese corporation, where lifelong employment was assumed. The result was an interview with relatively little structure, so that the interviewer and interviewee could talk more naturally and more deeply. The rate of employee turnover for the year for those who went through the structured interview process was zero, demonstrating validity of the selection process. On the other hand, we found unevenness in the correlations between job performance evaluations during On the Job Training (OJT) and those during the interview, indicating need for further research with longer frame of time for employee evaluations. PMID- 15747555 TI - [Comparison between sketch map and pointing methods for evaluating spatial cognitive ability of secondary school students with mental retardation]. AB - A study was conducted to clarify the spatial ability of secondary school students with mental retardation. Two Experiments-1 and 2-were carried out to test the students' spatial knowledge using a sketch map and a pointing task. In Experiment 1, 14 students (mean IQ and SD, 57.69 and 14.13 respectively) participated and were asked to draw sketch maps of their school, and their route from school to home, and to point to landmarks displayed in photos with their finger. Only five of 13 maps of the school, and four of 14 maps of the route home were drawn without heavy distortion. However, the results for pointing out landmarks were fairly good (mean angular error, 26.86). In Experiment 2, 10 students at a different school (mean IQ and SD, 35.4 and 10.3 respectively) participated and were asked to complete the same tasks. Although the sketch maps and pointing performance were not accurate, the difference in accuracy between landmarks inside and outside the school indicated that the students had a better grasp of spatial representation when space was familiar and limited. The difference in results between the sketch map and pointing tasks implies that the two tasks require different spatial representations and cognitive processes. PMID- 15747557 TI - [New coping-model of self-regulated learning]. AB - In a study of self-regulated learning study, we proposed a new coping model, in order to comprehensively treat emotional regulation and learning strategies. We also included meta-emotion variables in the model for intervention, so that we could discuss them in relation to intervention. We hypothesized specifically that achievement goals and meta-emotions influenced emotional reactions, and which in turn influenced coping strategies. A questionnaire study was conducted with 193 schoolchildren, and result seemed to support our new model. PMID- 15747556 TI - [Examining the causal model of depression: the relation between depressogenic schemata and depression]. AB - This study examines the causal relation between depressogenic schemata and depression. Three structural equation models were tested two times among 149 students during five months: (1) one-way causal relation from depressogenic schemata to depression, (2) one-way causal relation from depression to depressogenic schemata, (3) reciprocal relation between depressogenic schemata and depression. Results showed the third model is the most adequate among three models. It is possible that depressogenic schemata influences depression and depression also has some effects on depressogenic schemata. As a result of reviewing previous studies, it is thought that there is a continuity is between clinical and non-clinical groups. Therefore, the findings of this study is useful in understanding the clinical image of depression. PMID- 15747558 TI - [Influence of personality traits on collage works]. AB - The present study investigated whether personality traits may influence the outcome of collage works. In this study, 60 undergraduates were asked to fill Revised NEO Personality Inventory (NEO-PI-R) and generate collage works. The relations between the five factors of the NEO-PI-R (Neuroticism, Extraversion, Openness, Agreeableness, and Conscientiousness) and some evaluation measures of collage works (constructional features of collage works and characteristic behavior patterns in the process of their generation) were examined. Results indicated that several subscales of personality traits were substantially correlated with some indices of both two measures. These findings suggest that collage work may be a useful tool for psychological assessment. PMID- 15747559 TI - [Effects of emotion work on burnout and stress among human service professionals]. AB - This study examined the relationship between emotion work (Zapf, 2002) and burnout. One hundred and eighty-two human service professionals (nurses and caregivers) completed questionnaires. A factor analysis revealed that the concept of emotion work had four main factors: "Negative emotions display", "Positive emotions display", "Emotional dissonance", and "Sensitivity requirements". In addition, the hierarchical regression analyses revealed only the main effect of "Emotional dissonance" on psychological stress reaction, whereas the main effects of all emotion work variables and two interaction effects of those on burnout. These results suggested that burnout was distinguished from psychological stress reaction by the differences in its relationship to emotion work. PMID- 15747560 TI - [Structural equation modeling using ability parameters: analysis of the situation where item parameters have been estimated by item response theory]. AB - One of the problems with structural equation modeling (SEM) is that the estimation of measurement equation is not separated from the estimation of structural equation. The main aim of this study was to propose a new method to overcome that problem by using ability parameters estimated by item response theory (IRT) as data. According to IRT, the error variance of measurement equation can be easily computed as the reciprocal of the information function. By using the estimates of the error variance, we can fix all parameters in measurement equation and can separate the estimation of structural equation from that of measurement equation. This method also allows us to estimate relations among factor scores with improved precision, because the errors of estimating factor scores are taken into account. The article concludes with a simulation result for verifying the efficacy of this method and an actual numerical. PMID- 15747561 TI - [Construction of a stress reaction scale based on an item response model and examination of the developmental process of psychological stress reactions by using test characteristic curves]. AB - In this study, a stress reaction scale was constructed based on an Item Response Model, and the developmental process of psychological stress reactions was investigated by using test characteristic curves. Subjects consisted of 286 private college students and 234 national college students. The Job Stress Scale (JSS) was revised for college students, and used to assess psychological stress reaction. Graded response model of Item Response Theory (IRT) was used to estimate parameters. After estimating item and subject parameters, we assessed the precision of measurement and the fit of the model, and found both precision and fit to be satisfactory. Then we examined the test characteristic curves of each subscale to investigate the developmental process of psychological stress reactions of college students in comparison with company employees. PMID- 15747562 TI - [Effects of fundamental frequency and speech rate on impression formation]. AB - This study investigated the systematic relationship between nonverbal features of speech and personality trait ratings of the speaker. In Study 1, fundamental frequency (F0) in original speech was converted into five levels from 64% to 156.25%. Then 132 undergraduates rated each of the converted speeches in terms of personality traits. In Study 2 134 undergraduates similarly rated the speech stimuli, which had five speech rate levels as well as two F0 levels. Results showed that listener ratings along Big Five dimensions were mostly independent. Each dimension had a slightly different change profile over the five levels of F0 and speech rate. A quadratic regression equation provided a good approximation for each rating as a function of F0 or speech rate. The quadratic regression equations put together would provide us with a rough estimate of personality trait impression as a function of prosodic features. The functional relationship among F0, speech rate, and trait ratings was shown as a curved surface in the three-dimensional space. PMID- 15747563 TI - [Human efficiency for symmetry detection on 3D bumpy surfaces]. AB - Based on the ideal observer analysis, we investigated sampling properties of image information used by human visual system, for symmetrical pattern discrimination on 3D bumpy surface. There were three models of ideal observer (IO) to perform the task: 2D-IO using 2D projection image (i.e., retinal image), 2.5D-IO using image transformed to canonical view, and 3D-IO using recovered pattern image of 2D plane. We measured discrimination thresholds on the task for each IO model and subjects, and calculated human statistical efficiency relative to each ideal observer. The results indicated for the detection of a diagonal symmetry in the bumpy surface that human performance was similar to 3D-IO. This implies that human observers use the structure of the bumpy surface to detect the diagonal symmetry. PMID- 15747564 TI - [Validation of a Japanese research version of the Maslach Burnout Inventory General Survey]. AB - The purpose of this study was to validate Japanese research version of Maslach Burnout Inventory-General Survey (MBI-GS), a newly developed measure intended for persons of every occupation. The Japanese MBI-GS was administered to a sample of hospital workers. Exploratory factor analysis found three factors, as in original MBI-GS. Confirmatory factor analysis largely supported MBI-GS structure of three subscales, but the correlation between two subscales was unexpectedly high. To examine its construct validity, the subscale scores were then examined in relation to selected work characteristics. Conservation of resources theory was successful in its predictions of different patterns of effects among the correlates and three burnout subscales. The successful predictions suggested that meaning of each subscale was quite distinct. In all, our examination showed that Japanese MBI-GS assessed the same three dimensions as the original measure for human service workers. PMID- 15747565 TI - [Development of CAS (Communion-Agency Scale): a measurement of positive and negative aspects of gender personality]. AB - The purpose of this study was to construct a scale to measure both the positive and negative aspects of communion and agency, and to examine relationships among subscales. First, 264 undergraduates, both men and women, rated themselves on 40 items, and CAS (Communion-Agency Scale) was constructed, which consisted of four subscales with 6 items each. Second, 472 undergraduates, 303 women and 169 men, completed the scale, and were assigned, according to their scores on positive communion and agency, to one of four gender types: androgynous, masculine, feminine, and undifferentiated. Analyses of variance for negative communion and agency, with gender types (positive communion and agency) as independent variables, were performed. Factor analysis was also performed on the data. Results indicated that higher positive communion was associated with less negative agency, and higher positive agency with less negative communion. We discussed merits of measuring positive as well as negative aspects of communion and agency. PMID- 15747566 TI - [Helping behavior and psychosocial well-being in elderly people]. AB - This study examined relationship between everyday helping behaviors and psychosocial well-being in elderly people. Elderly people attending an institute for adult education (n=471), age ranged from 57 to 82, participated in the study. They completed questionnaires that assessed the following variables: everyday helping behaviors, psychological reactions to helping, and psychosocial well being, defined as self-esteem, moral, and social integration. The data indicated that psychological reactions resulting from. helping consisted of three components: "positive effects on the self and others," "negative effects on the self and others," and "positive effects on the self." The helper's psychological reactions to helping had a significant effect on self-esteem, moral, and social integration. There were gender differences in the effect of these variables. These results suggest that an affirmative helping experience in everyday life provides elderly people with a feeling of psychosocial well-being. PMID- 15747567 TI - [Marital relationship, occupational life, and subjective well-being of married people]. AB - This research investigated mental health of marreid people in relation to their marital relationship, occupational life, and household income. A questionnaire was administered to husbands and wives, either in middle age or child-rearing years, to measure their degree of satisfaction in marital relationship, occupational life, household income, and subjective well-being. Results showed that satisfaction in workplace for men, and additionally satisfaction in marital relationship of men in child-rearing years, strongly predicted their subjective well-being. As for women, however, the strong association with subjective well being was found for satisfaction in marital relationship, for those who were unemployed or employed part-time. The association was strongest for those who were in child-rearing years. Satisfaction in workplace was as important as satisfaction in marital relationship for women who were employed full-time. These findings suggested that satisfaction gained from what a person concentrated most of his/her energy on, explained his/her subjective well-being very well. PMID- 15747568 TI - [On-line processing mechanisms in text comprehension: a theoretical review on constructing situation models]. AB - This article reviewed research on construction of situation models during reading. To position variety of research in overall process appropriately, an unitary framework was devised in terms of three theories for on-line processing: resonance process, event-indexing model, and constructionist theory. Resonance process was treated as a basic activation mechanism in the framework. Event indexing model was regarded as a screening system which selected and encoded activated information in situation models along with situational dimensions. Constructionist theory was considered to have a supervisory role based on coherence and explanation. From a view of the unitary framework, some problems concerning each theory were examined and possible interpretations were given. Finally, it was pointed out that there were little theoretical arguments on associative processing at global level and encoding text- and inference information into long-term memory. PMID- 15747569 TI - How we do it: Virtual colonoscopy. PMID- 15747570 TI - Case of the month. Lipoma of the sigmoid colon. PMID- 15747571 TI - Case of the month. Necrotizing fasciitis. PMID- 15747572 TI - Computed tomography assessment of lung structure and function in pulmonary edema. AB - By definition pulmonary edema is an abnormal accumulation of water in the lung. Consequently, the computed tomography (CT) appearance of pulmonary edema reflects the sequence of this accumulation. In early hydrostatic pulmonary edema, CT shows vascular engorgement and peribronchovascular cuffing that increases with the severity of edema and that is associate in late stage, with consolidations. In acute respiratory distress syndrome (ARDS), CT shows the proportion of injured parenchyma and depicts associated alterations as parenchymal infiltrate and consolidation, pleural effusion, pneumothorax. These merely morphological findings can be complemented with data from objective CT analysis of the lung parenchyma. Indeed CT can assess lung water noninvasively. Correlated with hydrodynamic parameter, these objective measurements show that the increase of lung density parallels parenchymal fluid overload. These data also show that the occurrence of ground glass opacities can precede the hemodynamic evidence of edema. PMID- 15747573 TI - MDCT of the duodenum: technique and clinical applications. AB - Computed Tomography (CT) is useful for evaluation of both benign and malignant pathology involving the duodenum. CT can detect and stage duodenal malignancies as well as detect inflammatory conditions such as duodenitis. Careful CT technique is necessary to optimize detection of abnormalities. This includes the use of oral contrast, preferably water, as well as i.v. contrast. 3D imaging can improve disease detection and help with surgical planning. This article reviews the current role of CT in duodenal imaging, describes appropriate CT protocols and illustrates a wide variety of duodenal pathology. PMID- 15747574 TI - Primary pulmonary arterial hypertension presenting as diffuse micronodules on CT. AB - Primary pulmonary arterial hypertension is a rare lethal disease that typically presents radiographically with enlarged central pulmonary arteries, pruning of the peripheral vasculature, and cardiomegaly but clear lung fields. Although it is a disease of unknown etiology, primary PAH has been associated with anorexigen use. We present a case of pulmonary arterial hypertension in a woman with a history of fenfluramine and phentermine use who presented with diffuse micronodules on computed tomography scan. Lung biopsy confirmed the micronodules were radiographic manifestations of extensive diffuse plexogenic arterial lesions. This report represents an unusual radiographic presentation of anorexigen related pulmonary arterial hypertension, and to our knowledge, the first case reported as presenting with diffuse micronodules on high resolution computed tomography scan. PMID- 15747575 TI - Hepatic epitheloid hemangioendothelioma: multiphase CT appearance and correlation with pathology. AB - Epitheloid hepatic hemangioendothelioma was defined as a specific entity by Weiss and Enzinger in 1982. Histologically, this tumor originates from vascular endothelial cells and spindle cells. We report 3 cases of this rare neoplasm that presented to our institution over the past 3 years and briefly review the pathologic and imaging findings on MDCT (multiphase detector computed tomography). PMID- 15747576 TI - CT angiography: principles, techniques and study optimization using 16-slice multidetector CT with isotropic datasets and 3D volume visualization. AB - Significant advances in computed tomography (CT) scanner technology as well as 3D imaging software and hardware have resulted in amazing improvements in CT imaging of blood vessels. CT angiography has now been incorporated into daily practice and in may cases has replaced conventional angiography. CT angiography has also resulted in new applications for CT such as coronary angiography. This article discusses the current state of the art of 3D CT angiography and reviews the literature. Extensive case examples are included. PMID- 15747577 TI - Magnetic resonance imaging and proton magnetic resonance spectroscopy of intracranial epidermoid tumors. AB - Intracranial epidermoid tumors or cysts are considered benign lesions. Differentiation of epidermoid tumors from arachnoid cysts is important for appropriate patient care because the treatment is different for each lesion. Arachnoid cysts can appear very similar to epidermoid tumors on computed tomography (CT). Epidermoid tumors can grow in the cerebellopontine angle (CPA) cistern, the most common location of these lesions, resulting in trigeminal neuralgia and facial paralysis. Treatment for epidermoid tumors is exclusively surgery. Arachnoid cyst, on the other hand, is a benign condition that rarely produces symptoms. Recent advances in magnetic resonance imaging (MRI) have allowed more accurate imaging diagnosis of epidermoid tumors. This article reviews the recent advances in MRI using conventional T1W, post-contrast T1W, T2W, steady-state free precession imaging, fluid-attenuated inversion recovery (FLAIR), diffusion-weighted imaging (DWI), and proton magnetic resonance spectroscopy (MRS) in the imaging diagnosis of epidermoid tumors. PMID- 15747578 TI - Rheumatoid arthritis associated interstitial lung disease. AB - Rheumatoid arthritis (RA) is a systemic disease manifest as a symmetric polyarthritis usually in the setting of elevated autoantibodies (rheumatoid factor). This disease affects 1-2% of the world's population, most frequently in the 25-55 year old age group and has a female predominance (2.5:1). Nearly 50% of patients with RA demonstrate some type of extra-articular manifestation of the disease such as pleuritis, pleuropericarditis, vasculitis, pneumonitis, pulmonary fibrosis, scleritis or nodulosis. Pulmonary involvement in RA is common and can be due to the disease itself as well as to the therapies used to treat it. In fact, lung disease is the second most common cause of death, following infection, for patients with RA and has been reported to effect between 1-40% of patients. RA associated interstitial lung disease (ILD) is often subtle in onset, slowly progressive and of unclear etiology and response to treatment. This article aims to clarify the current clinical, radiographic and pathologic status of RA-ILD. PMID- 15747579 TI - Construction of a multi-functional cDNA library specific for mouse pancreatic islets and its application to microarray. AB - We have constructed a high-quality and multi-applicable cDNA library specific for mouse pancreatic islets. This is the first pancreatic islet cDNA library created using a recombination-based method, which can readily be converted into other applications including yeast two-hybrid and mammalian expression libraries. Based on sequence data of the library, we constructed a sequence database specific for mouse pancreatic islets. Among the 8882 non-redundant clones, 5799 were classified into specific functional categories using a classification system designed by the Gene Ontology Consortium, 10% of which were "molecular function unknown" genes. We also developed cDNA microarray membranes with 8108 non redundant clones. Analyses of expression profiles of three different cell lines and of MIN6 cells with or without overexpression of transcription factor NeuroD1 established the usefulness and applicability of our microarrays. The mouse pancreatic islet cDNA library, sequence database, set of clones, and microarrays developed in this study should be useful resources for studies of pancreatic islets and related diseases including diabetes mellitus. PMID- 15747580 TI - A 210-kb segment of tandem repeats and retroelements located between imprinted subdomains of mouse distal chromosome 7. AB - Mammalian genes subject to genomic imprinting often form clusters and are regulated by long-range mechanisms. The distal imprinted domain of mouse chromosome 7 is orthologous to the Beckwith-Wiedemann syndrome domain in human chromosome 11p15.5 and contains at least 13 imprinted genes. This domain consists of two subdomains, which are respectively regulated by an imprinting center. We here report the finished-quality sequence of a 0.6-Mb region encompassing the more centromeric subdomain. The sequence contains four imprinted genes (Ascl2/Mash2, Ins2, Igf2 and H19) and reveals previously unidentified CpG islands and tandem repeats, which may be features of imprinted genes. Most interestingly, a unique 210-kb segment consisting almost exclusively of tandem repeats and retroelements is identified. This segment, located between Th and Ins2, has features of heterochromatin-forming DNA and is highly methylated at CpG sites. The segment exhibits asynchronous replication on the parental chromosomes, a feature of the imprinted domains. We propose that this repeat segment could serve either as a boundary between the two subdomains or as a target for epigenetic chromatin modifications that regulate imprinting. PMID- 15747581 TI - Gene expression profiling identifies a set of transcripts that are up-regulated inhuman testicular seminoma. AB - Seminoma constitutes one subtype of human testicular germ cell tumors and is uniformly composed of cells that are morphologically similar to the primordial germ cells and/or the cells in the carcinoma in situ. We performed a genome-wide exploration of the genes that are specifically up-regulated in seminoma by oligonucleotide-based microarray analysis. This revealed 106 genes that are significantly and consistently up-regulated in the seminomas compared to the adjacent normal tissues of the testes. The microarray data were validated by semi quantitative RT-PCR analysis. Of the 106 genes, 42 mapped to a small number of specific chromosomal regions, namely, 1q21, 2p23, 6p21-22, 7p14-15, 12pll, 12p13, 12q13-14 and 22q12-13. This list of up-regulated genes may be useful in identifying the causative oncogene(s) and/or the origin of seminoma. Furthermore, immunohistochemical analysis revealed that the seminoma cells specifically expressed the six gene products that were selected randomly from the list. These proteins include CCND2 and DNMT3A and may be useful as molecular pathological markers of seminoma. PMID- 15747582 TI - Identification of transcripts expressed preferentially in hemocytes of Ciona intestinalis that can be used as molecular markers. AB - The immunity provided by ascidian hemocytes represents one prototype of innate immune function in vertebrates. However, there are currently no molecular markers of ascidian hemocytes. We accumulated a large number of ESTs of cDNAs derived from hemocytes of Ciona intestinalis, a cosmopolitan species of ascidian. By comparing these ESTs with those derived from other tissues and developmental stages of Ciona, we were able to extract 81 transcripts expressed abundantly and preferentially in hemocytes. Among them, the von Willebrand factor type A (vWA) like and complement 6 (C6)-like transcripts were found to be expressed almost exclusively in hemocytes, based on RT-PCR analysis and whole mount in situ hybridization. We propose that vWA-like and C6-like can be used as molecular markers for Ciona hemocytes. PMID- 15747583 TI - A simple and robust method for preparation of cDNA nylon microarrays. AB - DNA array technology has made remarkable progress in recent years and has become an indispensable tool in molecular biology research. However, preparing high quality custom-made DNA arrays at a reasonable cost is still an important concern because we cannot abandon the use of DNA array systems designed for specific purposes. To address these problems, we here report the use of rolling circle amplification products of cDNA plasmids dissolved in 80% formamide as DNA probes immobilized on a nylon membrane. First, because formamide is practically non volatile under ambient conditions and nucleic acids are easily dissolved in it, the use of formamide as a DNA solvent ensures that the DNA concentration of the solution will not change during arraying, which often takes several hours to a day depending on the number of DNA spots and arrays to produce. Secondly, the use of rolling circle amplification technology greatly reduced the labor needed to prepare the spotted DNA. The results in this study demonstrate that the introduction of these two modifications in preparation of nylon DNA array greatly improved its quality. PMID- 15747584 TI - [Antibacterial activity of oral cephems against various clinically isolated strains and evaluation of efficacy based on the pharmacokinetics/pharmacodynamics theory]. AB - We compared the antimicrobial activity of commercially available oral cephem agents, cefaclor (CCL), cefroxadine (CXD), cefdinir (CFDN), cefixime (CFIX), cefpodoxime (CPDX), cefteram (CFTM), cefcapene (CFPN), and cefditoren (CDTR), against Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyogenes, and ESBL-producing bacteria isolated from clinical materials in Kansai Medical University Hospital between 2002 and 2003. Based on the Pharmacokinetics/Pharmacodynamics (PK/PD) theory, we determined the concentration of each agent at which the time above MIC (TAM) value was 40% or more, and calculated the rate of efficacy against each type of bacteria. In S. pneumoniae strains, the MIC(50,80,90) values of CDTR were 0.25, 0.5, and 0.5 microg/ml, respectively, lower than those of the other agents, demonstrating the most potent antimicrobial activity. However, the efficacy rate for CDTR calculated based on the PK/PD theory was 58.5%. CFTM showed the highest efficacy rate (66.1%). In H. influenzae strains, the antimicrobial activity of CDTR was most potent, followed by that of CFTM and that of CFPN/CFIX. The MIC90 value of CDTR was lowest (0.25 microg/ml), followed by that of CFTM (0.5 microg/ml). The efficacy rate for CDTR was 100%. This result supports that CDTR frequently eradicates H. influenzae. In E. coli strains, the MIC90 values of the above agents, excluding CCL and CXD, ranged from 0.5 to 1 microg/ml. The antimicrobial activity of CFIX against K. pneumoniae was most potent, followed by that of CFDN/CPDX and that of CFTM. In ESBL-producing bacteria, most agents showed an MIC90 value of more than 4 microg/ml. In S. agalactiae and S. pyogenes strains, all of the agents showed satisfactory MIC values. In methi- cillin-sensitive Staphylococcus aureus (MSSA) strains, CFDN and CXD showed a high efficacy rate, whereas the efficacy rates for the other agents were low. The frequent use of oral agents has increased the number of cephem-resistant bacteria. ESBL-producing bacteria become highly resistant, and the presence or absence of response can be readily evaluated. However, when a mutation of penicillin-binding protein (PBP) occurs, drug resistance is less marked. Therefore, it is difficult to evaluate the treatment response in many cases. In S. pneumoniae strains, the efficacy rates for all of the agents were low in the evaluation using the PK/PD theory, suggesting that a dose higher than the standard dose should be established. Thus, in the future, the efficacy should be evaluated based on the PK/PD theory, appropriate antimicrobial treatment should be administered, and the administration method that does not increase the number of resistant bacteria must be established. PMID- 15747585 TI - [Antimicrobial susceptibility surveillance of Streptococcus pneumoniae and Haemophilus influenzae isolates to cefteram and other antimicrobial drugs during January 2003 to July 2004]. AB - The antimicrobial susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolates during January 2003 to July 2004 was determined to seven various antimicrobial drugs including cefteram (CFTM). The in vitro activities of these drugs against the fresh isolates were compared. The oral cephalosporins including CFTM were potently active against penicillin susceptible S. pneumoniae. The activity of CFTM and cefditoren was the most active among four oral cephalosporins. The susceptibilities of penicillin intermediate S. pneumoniae and penicillin resistant S. pneumoniae to antimicrobial agents were decreased. The MIC of CFTM was not beyond 4 microg/mL for any isolate of S. pneumoniae. The activity of CFTM was very high to beta-lactamase-negative and ampicillin susceptible H. influenzae isolates. These MIC against all isolates were 0.03 microg/mL or less. The MIC of CFTM was not beyond 1 microg/mL for any isolate of beta-lactamase-positive H. influenzae or beta-lactamase-negative-ampicillin resistant H. influenzae. In conclusion, CFTM exhibits a potent activity against fresh isolates of S. pneumoniae and H. influenzae, and has a potential of effectiveness in the infections. PMID- 15747587 TI - Multivariate parametric spatiotemporal models for county level breast cancer survival data. AB - In clustered survival settings where the clusters correspond to geographic regions, biostatisticians are increasingly turning to models with spatially distributed random effects. These models begin with spatially oriented frailty terms, but may also include further region-level terms in the parametrization of the baseline hazards or various covariate effects (as in a spatially-varying coefficients model). In this paper, we propose a multivariate conditionally autoregressive (MCAR) model as a mixing distribution for these random effects, as a way of capturing correlation across both the regions and the elements of the random effect vector for any particular region. We then extend this model to permit analysis of temporal cohort effects, where we use the term "temporal cohort" to mean a group of subjects all of whom were diagnosed with the disease of interest (and thus, entered the study) during the same time period (say, calendar year). We show how our spatiotemporal model may be efficiently fit in a hierarchical Bayesian framework implemented using Markov chain Monte Carlo (MCMC) computational techniques. We illustrate our approach in the context of county level breast cancer data from 22 annual cohorts of women living in the state of Iowa, as recorded by the Surveillance, Epidemiology, and End Results (SEER) database. Hierarchical model comparison using the Deviance Information Criterion (DIC), as well as maps of the fitted county-level effects, reveal the benefit of our approach. PMID- 15747586 TI - [New trends in research on group B streptococci (GBS) infections and sexual transmitted diseases]. AB - The adherence to and invasion of A549 cells by serotype VIII GBS (group B streptococci) were significantly greater than those of serotype III GBS for both the invasive strain and vaginal isolates. Serotype III strains of GBS stimulated significantly greater tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL 10), and interleukin-12 (IL-12) production than did serotype VIII strains. Interleukin-4 (IL-4) and interleukin-8 (IL-8) production in response to serotype VIII was significantly higher than in response to serotype III. On sexual transmitted diseases, orogenital contact has been common in both commercial sex workers (CSWs) and non-CSWs in Japanese women. Therefore, chlamydial infections in pharynx have been increasing in recent days. However, the treatment against Chlamydia trachomatis in the cases of pharyngeal infections should be longer than in the cases of uterine cervical infections. PMID- 15747588 TI - Comparison between two partial likelihood approaches for the competing risks model with missing cause of failure. AB - In many clinical studies where time to failure is of primary interest, patients may fail or die from one of many causes where failure time can be right censored. In some circumstances, it might also be the case that patients are known to die but the cause of death information is not available for some patients. Under the assumption that cause of death is missing at random, we compare the Goetgbebeur and Ryan (1995, Biometrika, 82, 821-833) partial likelihood approach with the Dewanji (1992, Biometrika, 79, 855-857) partial likelihood approach. We show that the estimator for the regression coefficients based on the Dewanji partial likelihood is not only consistent and asymptotically normal, but also semiparametric efficient. While the Goetghebeur and Ryan estimator is more robust than the Dewanji partial likelihood estimator against misspecification of proportional baseline hazards, the Dewanji partial likelihood estimator allows the probability of missing cause of failure to depend on covariate information without the need to model the missingness mechanism. Tests for proportional baseline hazards are also suggested and a robust variance estimator is derived. PMID- 15747589 TI - A distribution for multivariate frailty based on the compound Poisson distribution with random scale. AB - Frailty models are often used to model heterogeneity in survival analysis. The most common frailty model has an individual intensity which is a product of a random factor and a basic intensity common to all individuals. This paper uses the compound Poisson distribution as the random factor. It allows some individuals to be non-susceptible, which can be useful in many settings. In some diseases, one may suppose that a number of families have an increased susceptibility due to genetic circumstances. Then, it is logical to use a frailty model where the individuals within each family have some shared factor, while individuals between families have different factors. This can be attained by randomizing the Poisson parameter in the compound Poisson distribution. To our knowledge, this is a new distribution. The power variance function distributions are used for the Poisson parameter. The subsequent appearing distributions are studied in some detail, both regarding appearance and various statistical properties. An application to infant mortality data from the Medical Birth Registry of Norway is included, where the model is compared to more traditional shared frailty models. PMID- 15747590 TI - Marginal analysis for clustered failure time data. AB - Clustered failure time data are commonly encountered in biomedical research where the study subjects from the same cluster (e.g., family) share the common genetic and/or environmental factors such that the failure times within the same cluster are correlated. Two approaches that are commonly used to account for the intra cluster association are frailty models and marginal models. In this paper, we study the marginal proportional hazards model, where the structure of dependence between individuals within a cluster is unspecified. An estimation procedure is developed based on a pseudo-likelihood approach, and a risk set sampling method is proposed for the formulation of the pseudo-likelihood. The asymptotic properties of the proposed estimators are studied, and the related issues regarding the statistical efficiencies are discussed. The performances of the proposed estimator are demonstrated by the simulation studies. A data example from a child vitamin A supplementation trial in Nepal (Nepal Nutrition Intervention Project-Sarlahi, or NNIPS) is used to illustrate this methodology. PMID- 15747591 TI - A class of parametric dynamic survival models. AB - A class of parametric dynamic survival models are explored in which only limited parametric assumptions are made, whilst avoiding the assumption of proportional hazards. Both the log-baseline hazard and covariate effects are modelled by piecewise constant and correlated processes. The method of estimation is to use Markov chain Monte Carlo simulations: Gibbs sampling with a Metropolis-Hastings step. In addition to standard right censored data sets, extensions to accommodate interval censoring and random effects are included. The model is applied to two well known and illustrative data sets, and the dynamic variability of covariate effects investigated. PMID- 15747592 TI - Estimation in the positive stable shared frailty Cox proportional hazards model. AB - Shared frailty models are of interest when one has clustered survival data and when focus is on comparing the lifetimes within clusters and further on estimating the correlation between lifetimes from the same cluster. It is well known that the positive stable model should be preferred to the gamma model in situations where the correlated survival data show a decreasing association with time. In this paper, we devise a likelihood based estimation procedure for the positive stable shared frailty Cox model, which is expected to obtain high efficiency. The proposed estimator is provided with large sample properties and also a consistent estimator of standard errors is given. Simulation studies show that the estimation procedure is appropriate for practical use, and that it is much more efficient than a recently suggested procedure. The suggested methodology is applied to a dataset concerning time to blindness for patients with diabetic retinopathy. PMID- 15747593 TI - Pair chart test for an early survival difference. AB - The log-rank test is commonly used in comparing survival distributions between treatment and control groups in clinical trials. However, in many studies, the treatment is only effective at the early stage of the trial. Especially when the two survival curves cross, the log-rank test has a low statistical power to show the survival difference. We propose a test statistic for detecting such an early difference between the two treatment arms. The new test has an intuitive geometric interpretation based on a pair chart and is shown to have more power than the log-rank test when the treatment effect only appears in the early phase of the study. This advantage is evaluated for finite sample sizes in simulation studies. Finally, the proposed method is illustrated with a real data example of patients with gastric cancer. PMID- 15747594 TI - Multilevel mixed linear models for survival data. AB - For the analysis of correlated survival data mixed linear models are useful alternatives to frailty models. By their use the survival times can be directly modelled, so that the interpretation of the fixed and random effects is straightforward. However, because of intractable integration involved with the use of marginal likelihood the class of models in use has been severely restricted. Such a difficulty can be avoided by using hierarchical-likelihood, which provides a statistically efficient and fast fitting algorithm for multilevel models. The proposed method is illustrated using the chronic granulomatous disease data. A simulation study is carried out to evaluate the performance. PMID- 15747595 TI - Comparison of dewaterability and floc structure of synthetic sludge with activated sludge. AB - This work compared the appearance, dewaterability, floc size, fractal dimensions derived from free-settling test and light-scattering test, and detailed structural information of floc interior including porosity, Sierpinski carpet fractal dimension, and the fractal dimension of the pore boundaries of a synthetic sludge and an activated sludge. The present synthetic sludge could probe the change in filterability and in fractal dimensions extracted from small angle light scattering test and from free-settling test after conditioning, however, failed to reproduce the detailed structural information of sludge flocs. The use of specific synthetic sludge depends on the field of interest. For instance, the present synthetic sludge is not appropriate to simulate the intrafloc transport of solute owing to its incapability to reproduce the detailed floc structure or to its change after conditioning. PMID- 15747596 TI - The use of Fenton's system in the yeast industry wastewater treatment. AB - The paper presents the results of the research conducted with the use of hydrogen peroxide and iron (II) sulfate or chloride in the chemical pretreatment of Saccharomyces cerevisae yeast industry wastewater. It was found that the use of Fenton's system permitted a high reduction of sugar-like substances and total decolorizing of non-sugar compounds. The level of COD reduction depended on the amount and mutual proportions of COD:Fe(II):H2O2, as well as a type of the applied salt Fe(II). For iron concentrations: 1000-4000 mg l(-1) with molar excess [H2O2]:[Fe(II)] - 2-14:1 and reaction pH - 3.1-3.4, very high reproducibility of results and the COD reduction exceeding 75% were obtained. For this range of the reagent concentrations, the distribution of COD reduction values correlated with the equation: COD = - Ax4 + Bx3 - Cx2 + Dx - E (where: x = [H2O2]:[Fe(II)]). Additional neutralization with the use of lime milk made the secondary reduction of CODr(CaO) value possible, which resulted in the reduction of the total CODT above 90%. The method enabled us to consider the possibility of the preliminary chemical elimination of the wastewater load, which might increase the effectiveness of working wastewater treatment plants, especially in cases of continuous and occasional overloads above the level assumed by the project. PMID- 15747597 TI - Comparison of influence of free ammonia and dissolved oxygen on nitrite accumulation between suspended and attached cells. AB - The shortcut biological nitrogen removal (SBNR) hybrid (suspended cells combined with attached cells) process is an innovative technology that nitrosofies ammonium to nitrite and then denitrifies nitrite to nitrogen gas. Theoretically, this results in a 25% savings of the oxygen needed for nitrification and a 40% of savings in carbon source needed for denitrification. In this study, the influences of free ammonia (FA) and dissolved oxygen (DO) concentrations on nitrite accumulation were investigated to find the optimal operational factors for stable nitrite accumulation over a long period. The maximum specific utilization rates for ammonium (qa) and nitrite (qn) were determined for suspended and attached cells taken from a bench-scale SBNR reactor and a pilot scale livestock wastewater treatment plant reactor. For the ammonium and nitrite oxidations in both reactors, the attached cells were more resistant to the FA concentration, but were more significantly influenced by the DO concentration than the suspended cells. In addition, the effect of the DO concentration was more significant than that of the FA concentration for both types of cells from both reactors. In this SBNR hybrid system, a simultaneous manipulation of DO concentration (<1.5 mg l(-1)) and FA concentration (10-20 mg l(-1)) was required for maintaining high levels of nitrite accumulation. PMID- 15747598 TI - [Characterisation and environnemental impacts of olive oil wastewater disposal]. AB - The management of wastewater issued from olive oil extraction (locally named margine), continues to cause enormous problems due to the great quantities annually produced, and the contamination risks related to their high level of pollution and toxicity. The nearness of Agareb zone to the main olive oil production zone of Sfax and the existence of clayey outcrops are the main factors influencing the choice of this zone for the implantation of a site for the margine evaporation. The investigation and sampling works developed on the site have established the geological, hydrogeological, geotechnical and environmental characteristics. The permeable silty and sandy layers in the site favorized the infiltration of margine through and below the storage basins. This infiltration is particularly observed in the south part of the site where it has reached a depth of 4m at a distance of 20 m laterally. The chemical analysis of the substratum and infiltrated margine showed that the latter constituted a polluting source, its acidity may affect physical and geotechnical clay layer properties and causes pollutant migration in the subsoil. The wastewater infiltration in the underlying layers has caused the dissolution of salts and the modification of pH, nutrients contents and phenolic compound concentrations. The geotechnical investigations conducted on the site showed the instability of the embankments slopes separating the margine storage basins and the presence of sliding which may cause margine flow out from the storage basins and consequently the spreading of pollution on great areas. PMID- 15747600 TI - Seasonal changes in the structure of RBC heterotrophic biofilms. AB - Heterotrophic Biofilms, growing in the successive three treatment stages of the RBC reactor in a full-scale wastewater treatment plant in the summer of 2002, were investigated and compared to those growing in the fall of 2001. The seasonal changes in biofilm structure were studied in terms of biofilm thickness, dry solid density, internal pore distribution and patterns, and surface roughness. The comparison results indicated that due to the higher wastewater flow rate in the summer of 2002, these biofilms had thinner, denser and less rugged surface structure than the biofilms sampled in the fall of 2001; furthermore, they showed less heterogeneous intemal pore distribution. However, biofilms from both seasons showed some similarities: biofilms in the treatment stages 2 and 3 had similar structure; and their internal pore distribution and pore surface roughness didn't show any correlation with the biofilm depth. PMID- 15747599 TI - Biomass selection for optimal anaerobic treatment of olive mill wastewater. AB - This research was conducted to identify the most efficient biomass out of five different types of biomass sources for anaerobic treatment of Olive Mill Wastewater (OMW). This study was first focused on examining the selected biomass in anaerobic batch systems with sodium acetate solutions (control study). Then, the different types of biomass were tested with raw OMW (water-diluted) and with pretreated OMW by coagulation-flocculation using Poly Aluminum Chloride (PACl) combined with hydrated lime (Ca(OH)2). Two types of biomass from wastewater treatment systems of a citrus juice producing company "PriGat" and from a citric acid manufacturing factory "Gadot", were found to be the most efficient sources of microorganisms to anaerobically treat both sodium acetate solution and OMW. Both types of biomass were examined under different concentration ranges (1-40 g l(-1)) of OMW in order to detect the maximal COD tolerance for the microorganisms. The results show that 70-85% of COD removal was reached using Gadot biomass after 8-10 days when the initial concentration of OMW was up to 5 g l(-1), while a similar removal efficiency was achieved using OMW of initial COD concentration of 10 g l(-1) in 2-4 days of contact time with the PriGat biomass. The physico-chemical pretreatment of OMW was found to enhance the anaerobic activity for the treatment of OMW with initial concentration of 20 g l(-1) using PriGat biomass. This finding is attributed to reducing the concentrations of polyphenols and other toxicants originally present in OMW upon the applied pretreatment process. PMID- 15747601 TI - Ozonation of membrane bioreactor effluent for landfill leachate treatment. AB - Ozonation of leachate effluent obtained from a Membrane Bioreactor (MBR) process treating a medium-aged landfill leachate was investigated. The sequence of ammonia stripping, membrane bioreactor, and ozonation was used in the experiment. Ozonation of the MBR effluent showed reduction in Chemical Oxygen Demand (COD) and Total Organic Carbon (TOC) with an increase in five days Biochemical Oxygen Demand (BOD5) concentration. About 73% of COD could be removed at an ozone (O3) dosage of 4.2 mg O3 mg COD(-1). The biodegradability indicated by BOD5/COD ratio increased from 0.03 to 0.08 after ozonation at optimum ozone condition. While analyzing the molecular weight distribution, the fraction of molecular weight (MW) greater than 50 kDa of MBR effluent was transferred to MW 10-50 kDa (38%) at an ozone dosage of 810 mg O3. When O3 dosage was increased to 2,025 mg O3, a rapid change occurred in MW > 50 kDa fraction. This fraction was completely broken down into lower molecular weight fractions. This indicates partial degradation of landfill leachate by ozonation. The colour removal from the leachate was found to be greater than 95%. When the ozonated leachate was recirculated into the MBR system, along with an improvement in the performance of MBR, overall COD removal increased from 78% to 83%. Further, the ozone consumption decreased from 6.1 to 3.4 mg O3 mg COD(-1) removed, thus reducing the ozone requirement. PMID- 15747602 TI - Removal of copper and copper-binding organic ligands during potable water treatment. AB - The concentrations and chemical speciation of copper before and after potable water treatment have been examined using 3 techniques: a copper ion-selective electrode (Cu-ISE), chemiluminescence (CL) and anodic stripping voltammetry (ASV), together with water quality parameters colour, turbidity and dissolved organic carbon (DOC). As expected, only a small fraction of colour (6.6-11%), turbidity (6.7-14%) and DOC (20%) remained after treatment. The removal of Cu and Cu-binding natural organic matter (NOM) was more variable but still significant: 17-64% of total dissolved Cu and 19-61% of Cu-binding NOM (as determined by ASV) remained after treatment. Labile Cu levels, as measured by CL, were 0-23% after treatment. The Cu-ISE titrations showed significant Cu-binding in raw water in the concentration range 10(-7) to 10(-6) mol l(-1) Cu2+, but very little binding after treatment. The results indicate that conventional water treatment has a significant effect on both total Cu levels and Cu-binding NOM, with implications for Cu2+ speciation and bio-availability, particularly in reticulation systems containing copper pipes. PMID- 15747603 TI - Community changes during start-up in methanogenic bioreactors exposed to increasing levels of ammonia. AB - To investigate the methanogenic population dynamics during the start-up under gradually increasing free ammonia levels, five mesophilic UASB reactors seeded with different sludges were operated for 140 days. Changes in the methanogenic community investigated by using 16S rDNA/rRNA based molecular methods such as denaturing gradient gel electrophoresis (DGGE), fluorescent in-situ hybridization (FISH), cloning and DNA sequencing. Free ammonia nitrogen (FAN) in the reactors was gradually increased from 50 to 130 mg l(-1) by increasing total ammonia nitrogen (TAN) from 1000 to 2500 mg l(-1) in the feed and keeping reactor pH at 7.7 in the active zone. Even at highest FAN level, COD and VFA removal efficiencies above 90 and 98% were obtained, respectively. However, Methanosaeta related species mainly prevailing in seed sludges were substituted for Methanosarcina as the abundant methanogens although reactor performances were almost comparable and constant. PMID- 15747604 TI - Determination of environmental factors influencing methane oxidation in a sandy landfill cover soil. AB - It is advantageous to use coarse soils as landfill cover because they allow better aeration of the biologically active zone. In this study, therefore, patterns of methane oxidation were investigated under various environmental conditions including soil moisture content, temperature, and the addition of NH4+ in a sandy landfill cover soil. The kinetics of CH4 oxidation was also studied at different moisture contents and temperatures. Soil moisture content of 10% (wt/wt) resulted in the maximum CH4 oxidation rate (19.2-22.4 nmol gsoil DW(-1) min(-1)). A Vmax value was not significantly different when the moisture content was more than 10%, but a Km value increased from 5.23 to 75.24 microM as the moisture content increased. The ratio of Vmax to Km was the highest at 10% moisture content. The CH4 oxidation rate increased as the incubation temperature increased, and Q10 values and optimum temperature were determined to be 2.57-2.69 and 30 degrees C, respectively. Both Vmax and Km values decreased at the temperatures below and above 30 degrees C. The addition of various levels of NH4+ resulted in increased or decreased CH4 oxidation rates, however, the initiation of appreciable CH4 oxidation was delayed with increasing amounts of NH4+ application in all samples tested. Among the environmental variables tested, moisture content control seems to be the most important and an efficient means of managing methane oxidation when sandy soils are used in landfill cover. PMID- 15747605 TI - The use of a biofilter for reducing off-gas odour from an industrial fermentation process. AB - This study evaluated the performance of a lab-scale, closed bed biofilter for the removal of odour in off-gas released from an industrial fermentation facility. This off-gas was emitted from the facility periodically after the sterilization of fermentation medium. The lab scale biofilter was operated for over two months, totalling 30 medium sterilizations. The biofilter was subjected to a shock odour load for each sterilization cycle and to two airflow conditions: the fermentation off-gas and compressed room air, which was cooler and drier than the fermentation off-gas. The biofilter was effective in removing odour under shock loading and variable operating conditions (temperature and relative humidity). An odour reduction rate of 72% was achieved immediately after medium sterilization when odour levels were highest (32 800 OU m(-3)). The filter had an odour removal efficiency of 61% and 67% for 24 h and 50 h after sterilization, respectively. PMID- 15747606 TI - Characterization and ultrafiltration of semiconductor indium phosphide (InP) wastewater for recycling. AB - This research work investigated the physical and chemical properties of a new type of wastewater produced from the semiconductor industry. The wastewater generated from indium phosphide (InP) wafer backgrinding and sawing processes was characterized in term of its particle size distribution (PSD), zeta potential, suspended and dissolved solids, total organic carbon, and turbidity. The wastewater contained high concentration of fine InP dusts with a size ranging from 0.07 - 1.44 mm. In spite of its high concentration of suspended solids resulting in high turbidity up to 371 NTU, the wastewater contained very low organic matters (TOC < 2.2 mg l(-1)) and other inorganic impurities (SO4(2-) < 0.21 mg l(-1) and Na+ < 0.16 mg l(-1)). Based on the experimental data collected, the treatment technologies using chemical precipitation and ultrafiltration were applied to the wastewater. Both processes could effectively remove InP particles from the wastewater, however the coagulants in chemical precipitation introduced other ionic contents into the process resulting in difficulties of water recycling in the later stage. In comparison, ultrafiltration was more promising for InP wastewater treatment and recycling. Based on the results of this study, a full-scale UF system was built in a local semiconductor plant and it has successfully reclaimed water from the InP wastes for the past six months without any quality issue being raised. PMID- 15747607 TI - Epidemiology of osteoarthritis. AB - The majority of adults > 55 years of age show radiographic evidence of osteoarthritis; however, determining definitive prevalence and incidence rates of knee osteoarthritis and hip osteoarthritis poses methodologic problems. Several factors play a role in osteoarthritis risk, including age, gender, genetics, behavioral influences, and ethnicity. Studies cited compare data from the Framingham Osteoarthritis Study with data from studies performed in Asian and European populations. A number of broad trends stand out, including higher prevalence in advanced age for both hip and knee osteoarthritis, with higher prevalence and incidence in women than in men. PMID- 15747609 TI - Treatment options for osteoarthritis. AB - Osteoarthritis is the most common form of arthritis and can be a major source of disability. Many older patients continue to be active in sports-related activities. Therefore, treating an active patient with osteoarthritis is becoming more common. Physical therapy and weight loss continue to play a central role in the treatment of patients with osteoarthritis. Nonsteroidal anti-inflammatory drugs decrease inflammation and swelling, which makes physical therapy more effective. Intra-articular corticosteroids have been used for the treatment of osteoarthritis. However, no guidelines exist for the administration of corticosteroids and they can be associated with increased risk of tendon rupture and infection. Viscosupplementation has gained popularity in the treatment of osteoarthritis of the knee. Intra-articular injections of hyaluronic acid have been shown to decrease pain and improve functional outcomes. PMID- 15747608 TI - Etiology and pathophysiology of osteoarthritis. AB - Acute or chronic insult, including normal wear and tear, age, obesity, and joint injury, may initiate an imbalance between matrix synthesis and matrix degradation in healthy cartilage that promotes chondral loss and prevents cartilage self repair. The structure of healthy cartilage and the pathophysiological mechanisms of its degradation are described, followed by descriptions of endogenous and exogenous factors believed to be involved in the progressive course of osteoarthritis. Studies cited include research from the community of sports medicine. PMID- 15747610 TI - Osteoarthritis case study. PMID- 15747612 TI - [Androgen deficiency in the ageing male: a neuro-endocrine disease]. PMID- 15747611 TI - Future treatment of osteoarthritis. AB - Osteoarthritis represents an advanced stage of disease progression caused in part by injury, loss of cartilage structure and function, and an imbalance in inflammatory and noninflammatory pathways. The burden of this disease will increase in direct proportion to the increase in the older adult population. Research on current and experimental treatment protocols are reviewed, including the effect of hyaluronic acid in both in vitro and in vivo studies, autologous chondrocyte and osteochondral plug implantation, and gene therapy. Disease modifying osteoarthritis drugs and in vivo studies of glucosamine and chondroitin sulfate are reviewed. PMID- 15747613 TI - [Epidemiology of the ageing male]. PMID- 15747614 TI - [Physiology of androgens in adult men]. PMID- 15747615 TI - [Pathophysiology of androgen deficiency in the ageing male]. PMID- 15747616 TI - [Drug interactions with the androgen balance of adult men]. PMID- 15747617 TI - [Androgens, the ageing male and gynaecomastia]. PMID- 15747618 TI - [Action of androgens on fat distribution and metabolic balance]. PMID- 15747619 TI - [Androgens, skeletal muscle and muscle exercise]. PMID- 15747620 TI - [Androgens and bone metabolism]. PMID- 15747621 TI - [Androgens and the cardiovascular system]. PMID- 15747622 TI - [Androgen deficiency and sleep apnoea]. PMID- 15747623 TI - [Androgens and prostate: basic research and clinical data]. PMID- 15747624 TI - [Androgens, the ageing male and fertility]. PMID- 15747625 TI - [Androgens, behaviour and cognition]. PMID- 15747626 TI - [Testosterone and male sexuality: basic research and clinical data]. PMID- 15747627 TI - [Androgens, the ageing male and autonomic nervous system]. PMID- 15747628 TI - [Androgens and haematopoiesis]. PMID- 15747629 TI - [Androgens and immunity]. PMID- 15747630 TI - [Androgens and ageing of the skin]. PMID- 15747631 TI - [Age-related androgen deficiency: diagnostic work-up]. PMID- 15747632 TI - [Androgen replacement therapy in androgen deficiency in the ageing male]. PMID- 15747633 TI - [Is there a place for DHEA, GH and melatonin in the management of age-related androgen deficiency?]. PMID- 15747634 TI - [Treatment guidelines for the androgen deficiency in the ageing male]. PMID- 15747635 TI - Accelerated unsteady flow line integral convolution. AB - Unsteady flow line integral convolution (UFLIC) is a texture synthesis technique for visualizing unsteady flows with high temporal-spatial coherence. Unfortunately, UFLIC requires considerable time to generate each frame due to the huge amount of pathline integration that is computed for particle value scattering. This paper presents Accelerated UFLIC (AUFLIC) for near interactive (1 frame/second) visualization with 160,000 particles per frame. AUFLIC reuses pathlines in the value scattering process to reduce computationally expensive pathline integration. A flow-driven seeding strategy is employed to distribute seeds such that only a few of them need pathline integration while most seeds are placed along the pathlines advected at earlier times by other seeds upstream and, therefore, the known pathlines can be reused for fast value scattering. To maintain a dense scattering coverage to convey high temporal-spatial coherence while keeping the expense of pathline integration low, a dynamic seeding controller is designed to decide whether to advect, copy, or reuse a pathline. At a negligible memory cost, AUFLIC is 9 times faster than UFLIC with comparable image quality. PMID- 15747636 TI - Barycentric parameterizations for isotropic BRDFs. AB - A bidirectional reflectance distribution function (BRDF) is often expressed as a function of four real variables: two spherical coordinates in each of the the "incoming" and "outgoing" directions. However, many BRDFs reduce to functions of fewer variables. For example, isotropic reflection can be represented by a function of three variables. Some BRDF models can be reduced further. In this paper, we introduce new sets of coordinates which we use to reduce the dimensionality of several well-known analytic BRDFs as well as empirically measured BRDF data. The proposed coordinate systems are barycentric with respect to a triangular support with a direct physical interpretation. One coordinate set is based on the BRDF model proposed by Lafortune. Another set, based on a model of Ward, is associated with the "halfway" vector common in analytical BRDF formulas. Through these coordinate sets we establish lower bounds on the approximation error inherent in the models on which they are based. We present a third set of coordinates, not based on any analytical model, that performs well in approximating measured data. Finally, our proposed variables suggest novel ways of constructing and visualizing BRDFs. PMID- 15747637 TI - A multiresolution representation for massive meshes. AB - We present a new external memory multiresolution surface representation for massive polygonal meshes. Previous methods for building such data structures have relied on resampled surface data or employed memory intensive construction algorithms that do not scale well. Our proposed representation combines efficient access to sampled surface data with access to the original surface. The construction algorithm for the surface representation exhibits memory requirements that are insensitive to the size of the input mesh, allowing it to process meshes containing hundreds of millions of polygons. The multiresolution nature of the surface representation has allowed us to develop efficient algorithms for view-dependent rendering, approximate collision detection, and adaptive simplification of massive meshes. The empirical performance of these algorithms demonstrates that the underlying data structure is a powerful and flexible tool for operating on massive geometric data. PMID- 15747638 TI - Spatial domain wavelet design for feature preservation in computational data sets. AB - High-fidelity wavelet transforms can facilitate visualization and analysis of large scientific data sets. However, it is important that salient characteristics of the original features be preserved under the transformation. We present a set of filter design axioms in the spatial domain which ensure that certain feature characteristics are preserved from scale to scale and that the resulting filters correspond to wavelet transforms admitting in-place implementation. We demonstrate how the axioms can be used to design linear feature-preserving filters that are optimal in the sense that they are closest in L2 to the ideal low pass filter. We are particularly interested in linear wavelet transforms for large data sets generated by computational fluid dynamics simulations. Our effort is different from classical filter design approaches which focus solely on performance in the frequency domain. Results are included that demonstrate the feature-preservation characteristics of our filters. PMID- 15747639 TI - A statistical wisp model and pseudophysical approaches for interactive hairstyle generation. AB - This paper presents an interactive technique that produces static hairstyles by generating individual hair strands of the desired shape and color, subject to the presence of gravity and collisions. A variety of hairstyles can be generated by adjusting the wisp parameters, while the deformation is solved efficiently, accounting for the effects of gravity and collisions. Wisps are generated employing statistical approaches. As for hair deformation, we propose a method which is based on physical simulation concepts, but is simplified to efficiently solve the static shape of hair. On top of the statistical wisp model and the deformation solver, a constraint-based styler is proposed to model artificial features that oppose the natural flow of hair under gravity and hair elasticity, such as a hairpin. Our technique spans a wider range of human hairstyles than previously proposed methods and the styles generated by this technique are fairly realistic. PMID- 15747640 TI - Geometry-aware bases for shape approximation. AB - We introduce a new class of shape approximation techniques for irregular triangular meshes. Our method approximates the geometry of the mesh using a linear combination of a small number of basis vectors. The basis vectors are functions of the mesh connectivity and of the mesh indices of a number of anchor vertices. There is a fundamental difference between the bases generated by our method and those generated by geometry-oblivious methods, such as Laplacian-based spectral methods. In the latter methods, the basis vectors are functions of the connectivity alone. The basis vectors of our method, in contrast, are geometry aware since they depend on both the connectivity and on a binary tagging of vertices that are "geometrically important" in the given mesh (e.g., extrema). We show that, by defining the basis vectors to be the solutions of certain least squares problems, the reconstruction problem reduces to solving a single sparse linear least-squares problem. We also show that this problem can be solved quickly using a state-of-the-art sparse-matrix factorization algorithm. We show how to select the anchor vertices to define a compact effective basis from which an approximated shape can be reconstructed. Furthermore, we develop an incremental update of the factorization of the least-squares system. This allows a progressive scheme where an initial approximation is incrementally refined by a stream of anchor points. We show that the incremental update and solving the factored system are fast enough to allow an online refinement of the mesh geometry. PMID- 15747641 TI - Sharpen&Bend: recovering curved sharp edges in triangle meshes produced by feature-insensitive sampling. AB - Various acquisition, analysis, visualization, and compression approaches sample surfaces of 3D shapes in a uniform fashion without any attempt to align the samples with sharp edges or to adapt the sampling density to the surface curvature. Consequently, triangle meshes that interpolate these samples usually chamfer sharp features and exhibit a relatively large error in their vicinity. We present two new filters that improve the quality of these resampled models. EdgeSharpener restores the sharp edges by splitting the chamfer edges and forcing the new vertices to lie on intersections of planes extending the smooth surfaces incident upon these chamfers. Bender refines the resulting triangle mesh using an interpolating subdivision scheme that preserves the sharpness of the recovered sharp edges while bending their polyline approximations into smooth curves. A combined Sharpen&Bend postprocessing significantly reduces the error produced by feature-insensitive sampling processes. For example, we have observed that the mean-squared distortion introduced by the SwingWrapper remeshing-based compressor can often be reduced by 80 percent executing EdgeSharpener alone after decompression. For models with curved regions, this error may be further reduced by an additional 60 percent if we follow the EdgeSharpening phase by Bender. PMID- 15747642 TI - Camera-based calibration techniques for seamless multiprojector displays. AB - Multiprojector, large-scale displays are used in scientific visualization, virtual reality, and other visually intensive applications. In recent years, a number of camera-based computer vision techniques have been proposed to register the geometry and color of tiled projection-based display. These automated techniques use cameras to "calibrate" display geometry and photometry, computing per-projector corrective warps and intensity corrections that are necessary to produce seamless imagery across projector mosaics. These techniques replace the traditional labor-intensive manual alignment and maintenance steps, making such displays cost-effective, flexible, and accessible. In this paper, we present a survey of different camera-based geometric and photometric registration techniques reported in the literature to date. We discuss several techniques that have been proposed and demonstrated, each addressing particular display configurations and modes of operation. We overview each of these approaches and discuss their advantages and disadvantages. We examine techniques that address registration on both planar (video walls) and arbitrary display surfaces and photometric correction for different kinds of display surfaces. We conclude with a discussion of the remaining challenges and research opportunities for multiprojector displays. PMID- 15747643 TI - A practical approach to spectral volume rendering. AB - To make a spectral representation of color practicable for volume rendering, a new low-dimensional subspace method is used to act as the carrier of spectral information. With that model, spectral light material interaction can be integrated into existing volume rendering methods at almost no penalty. In addition, slow rendering methods can profit from the new technique of postillumination-generating spectral images in real-time for arbitrary light spectra under a fixed viewpoint. Thus, the capability of spectral rendering to create distinct impressions of a scene under different lighting conditions is established as a method of real-time interaction. Although we use an achromatic opacity in our rendering, we show how spectral rendering permits different data set features to be emphasized or hidden as long as they have not been entirely obscured. The use of postillumination is an order of magnitude faster than changing the transfer function and repeating the projection step. To put the user in control of the spectral visualization, we devise a new widget, a "light-dial," for interactively changing the illumination and include a usability study of this new light space exploration tool. Applied to spectral transfer functions, different lights bring out or hide specific qualities of the data. In conjunction with postillumination, this provides a new means for preparing data for visualization and forms a new degree of freedom for guided exploration of volumetric data sets. PMID- 15747644 TI - Warp sculpting. AB - The task of computer-based free-form shape design is fraught with practical and conceptual difficulties. Incorporating elements of traditional clay sculpting has long been recognized as a means of shielding the user from these complexities. We present warp sculpting, a variant of spatial deformation, which allows deformations to be initiated by the rigid body transformation or uniform scaling of volumetric tools. This is reminiscent of a tool imprinting, flexing, and molding clay. Unlike previous approaches, the deformation is truly interactive. Tools, encoded in a distance field, can have arbitrarily complex shapes. Although individual tools have a static shape, several tools can be applied simultaneously. We enhance the basic formulation of warp sculpting in two ways. First, deformation is toggled to automatically overcome the problem of "sticky" tools, where the object's surface clings to parts of a tool that are moving away. Second, unlike many other spatial deformations, we ensure that warp sculpting remains foldover-free and, hence, prevent self-intersecting objects. PMID- 15747645 TI - Correcting interperspective aliasing in autostereoscopic displays. AB - An image presented on an autostereoscopic system should not contain discontinuities between adjacent views. A viewer should experience a continuous scene when moving from one view to the next. If corresponding points in two perspectives do not spatially abut, a viewer will experience jumps in the scene. This is known as interperspective aliasing. Interperspective aliasing is caused by object features far away from the stereoscopic screen being too small, which results in visual artifacts. By modeling a 3D point as a defocused image point, we can adapt Fourier analysis to devise a depth-dependent filter kernel that allows filtering of a stereoscopic 3D image. For synthetic 3D data, we use a simpler approach, which is to smear the data by a distance proportional to its depth. PMID- 15747646 TI - Periodontal antimicrobials--finding the right solutions. AB - Strengthened by promising research data and commercial backing, interest in the field of anti-infective periodontal therapy is rapidly expanding. Management of the periodontal microbiota with antibiotic drugs and antiseptic agents in conjunction with mechanical debridement seems to be more effective than mechanical therapy alone, at least in the treatment of advanced periodontal disease. The choice of a periodontal chemotherapeutic regimen requires an understanding of the usual infecting flora, available antimicrobial agents, and pathogen susceptibility patterns. Systemic administration of combinations of metronidazole and either amoxicillin or ciprofloxacin has been widely used with great success; however the presence of subgingival yeasts and resistant bacteria can be a problem in some periodontitis patients. Valuable antiseptic agents for subgingival application include 10% povidone-iodine for professional use and 0.1 0.5% sodium hypochlorite for patient self-care. These antiseptics have significantly broader spectra of antimicrobial action, are less likely to induce development of resistant bacteria and adverse host reactions, and are considerably less expensive than commercially available antibiotics in controlled release devices. In practice, mechanical debridement combined with subgingival povidone-iodine application in the dental office and sodium hypochlorite irrigation for patient self-care are valuable antimicrobial remedies in the treatment of virtually all types of periodontal disease. Management of moderate to severe periodontitis may require additional systemic antibiotic and/or surgical treatment. PMID- 15747647 TI - Development and evaluation of a self-report measure for identifying type and use of removable partial dentures. AB - AIM: To develop and validate a self-complete questionnaire suitable for use in primary dental care settings. DESIGN: A questionnaire survey. PARTICIPANTS: The research was undertaken in primary dental care (general dental practices, Newcastle upon Tyne, UK) and two teaching hospitals (Newcastle Dental Hospital, UK; University Dental School and Hospital, Cork, Ireland). A self-assessment of numbers of teeth and removable partial dentures questionnaire was devised and distributed to 137 patients in the three settings. Following completion of the questionnaire, each patient's dentist then examined them and completed the same questionnaire. The two sets of information were then compared. In the second phase of the study, 37 subjects completed the questionnaire in their own home and their dentist completed the questionnaire at a subsequent clinical examination. RESULTS: There was excellent agreement regarding counting of teeth [80%, Kappa = 0.79, s.e. 0.04 for maxillary teeth and 77%, Kappa = 0.68, s.e. 0.03 for mandibular teeth]. There was also good agreement between dentists and patients in terms of positioning of spaces in the dental arch (79.6-92.6%). Subjects in the study were also able to accurately identify the type of denture they had (either acrylic or cobalt-chromium based). These findings were confirmed in the second phase of the study. CONCLUSIONS: Agreement between dentists and patients was very good. This self-report measure could be of use when gathering data for studies of partially dentate patients. PMID- 15747648 TI - Tooth loss in a sub-urban Nigerian population: causes and pattern of mortality revisited. AB - AIM: To determine the current causes and pattern of tooth loss in Ile-Ife Nigeria. DESIGN: A retrospective review of records of dental extraction patients. SETTING: The dental hospitals of the Obafemi Awolowo University Teaching Hospital situated in South Western Nigeria. PARTICIPANTS: All patients that had dental extractions between January 1996 and December 2002. METHOD: Patients' case records were analysed for demographics, reason(s) for dental extraction, tooth/teeth extracted, method of extraction and complication(s). MAIN OUTCOME MEASURE: Reasons for tooth extraction and the pattern of tooth loss. RESULTS: 6348 (12.3%) of hospital attendees aged 4-102 years (mean 35 +/- 16.8 years) had extraction of 8338 teeth. A statistically significant female preponderance was observed. Dental caries was the leading cause of tooth loss (56.4%) followed by periodontal disease (24.6%). This shows a reversal of a trend reported in a previous study in the same location. Over half (69.2%) of the extracted teeth were molars (mostly mandibular). Only 4.1% extractions were trans alveolar. 3.9% were complicated by dry socket. CONCLUSION: 12.3% of the hospital attendees had undergone dental extraction. Despite previous warnings of a steady rise in dental caries, it has become the leading cause of tooth loss in our hospital. The urgent need to institute standardised preventive measures was highlighted. PMID- 15747649 TI - Prevalence and severity of dental caries in Costa Rican schoolchildren: results of the 1999 national survey. AB - In 1987, Costa Rica implemented a comprehensive national salt fluoridation programme using sodium fluoride (225-275mg F/kg salt). AIM: To describe dental caries prevalence and severity in Costa Rican children in 1999. METHODS: Eight calibrated examiners (inter-examiner Kappa = 0.70 or higher) recorded information on dental caries, treatment needs, enamel fluorosis, and dentofacial anomalies for schoolchildren aged 6-8,12, and 15-years (N=3758). The survey utilised a multistrata probability sample with fixed allocation to represent seven regions of the country. RESULTS: The overall mean dmft for age 6-8 years was 3.32. The DMFT for age 12 was 2.46 and for age 15 was 4.37. Regional differences were observed; for example, the DMFT at age 12 years ranged from 1.93 to 3.86. Compared with pre-fluoridation data collected in 1984, schoolchildren aged 12 years, experienced a 28 per cent decrease in caries prevalence (100 per cent to 72 per cent) and a 73 per cent decrease in severity (DMFT from 9.13 to 2.46, representing an 8.3 per cent compound annual per cent reduction). CONCLUSIONS: Between 1984 and 1999, Costa Rican schoolchildren experienced substantial reductions in caries prevalence and severity. Many factors may be involved in this decline, but the most important appears to be exposure to fluoridated salt. PMID- 15747650 TI - Factors associated with perceived oral health status in older individuals. AB - AIM: To assess factors associated with perception of oral health in older individuals. DESIGN: A cross sectional study. SETTING: A densely populated urban area in Sri Lanka. PARTICIPANTS: 585 older adults aged 60 years and above of which 475 were living at home and 110 in institutions. The present analysis is limited to 235 subjects who were subjected to a clinical oral examination. METHOD: The data were collected by means of an interviewer-administered questionnaire and a clinical oral examination. RESULTS: Overall, 48% of the dentate and 42% of the edentate perceived their oral health as poor. The final model of the hierarchical logistic regression analysis for the dentate revealed that presence of retained roots, mobile teeth, >20 missing teeth and perceived need for dental care were significantly associated with poor perceived oral health status. For the edentate, perceived need for dental care, loss of taste sensation and difficulty in eating were significantly associated with poor perceived oral health status. CONCLUSIONS: Factors associated with perceived oral health status differed between the dentate and the edentate. Clinical oral health indicators emerged as significant predictors of perceived oral health status in the dentate although the explanatory power of these indicators on perception of oral health was low. PMID- 15747651 TI - Conservative restoration with resin composites of a case of amelogenesis imperfecta. AB - Amelogenesis imperfecta (AI) is an inherited enamel dysplasia involving both dentitions with no other systemic effects. The hereditary pattern is autosomal or X-related dominant or recessive. Its prevalence is approximately 1:14,000 1:16,000. It can be classified as hypocalcified, hypoplastic and hypomaturated according to clinical, radiological, histological and hereditary findings. This study presents a case of hypomaturated type AI in a 16-year-old young man that was successfully treated with different types of resin composites. The patient was regularly recalled during the one-year postoperative period. Radiographic and clinical examinations at recall revealed no evidence of complications associated with the restored teeth or their supporting structures. PMID- 15747652 TI - Cutting edge research that will impact future oral health care. PMID- 15747653 TI - West Midlands vet to be struck off. PMID- 15747654 TI - International disease surveillance. International disease monitoring, October to December 2004. PMID- 15747655 TI - Tissue distribution of bovine spongiform encephalopathy infectivity in Romney sheep up to the onset of clinical disease after oral challenge. AB - Sixty Romney sheep of three prion protein genotypes were dosed orally at six months of age with an inoculum prepared from the brains of cattle clinically affected with BSE, and 15 sheep were left undosed as controls. They were randomly assigned within genotype to groups and were sequentially euthanased and examined postmortem at intervals of six or 12 months, depending on their predicted susceptibility. Tissue pools prepared from the three, four or five dosed animals in each group were inoculated into groups of 20 RIII mice as a bioassay for infectivity. Separate inocula were prepared from the matched control sheep killed at each time. In the ARQ/ARQ sheep killed four months after inoculation, infectivity was detected in the Peyer's patch tissue pool, and at 10 months it was detected in the spleen pool; from 16 months, infectivity was detected in a range of nervous and lymphoreticular tissues, including the spinal cord pool, distal ileum excluding Peyer's patches, liver, Peyer's patches, mesenteric and prescapular lymph nodes, spleen, tonsil and cervical thymus. No infectivity was detected in the tissue pools from the ARQ/ARR and ARR/ARR sheep killed 10 months or 22 months after infection. PMID- 15747656 TI - Comparison of sodium bicarbonate and carbicarb for the treatment of metabolic acidosis in newborn calves. AB - Carbicarb (an equimolar mixture of sodium bicarbonate and sodium carbonate) was compared with sodium bicarbonate alone for the treatment of acidosis in newborn calves: 25 of 49 calves with a blood pH at birth of less than 7-2 and a base deficit of less than -3 mmol/litre were treated intravenously with sodium bicarbonate and 24 were treated with carbicarb. The doses were calculated on the basis of the base deficit in a blood sample taken 10 minutes after birth, and further blood samples were taken immediately after the treatment and 30 and 60 minutes after the treatment for the determination of acid-base status, blood gases and haematological and biochemical variables. Both treatments resulted in a significant increase in blood pH, but there was no difference between them. The mean (sd) blood pH before treatment was 7.09 (0.02) and after treatment it was 7.28 (0.01). There was no increase in the partial pressure of carbon dioxide after treatment with either sodium bicarbonate or carbicarb. Both treatments were associated with an increase in sodium concentration and decreases in the total erythrocyte count, haematocrit and haemoglobin concentration. PMID- 15747658 TI - Counting nematode eggs in equine faecal samples. PMID- 15747657 TI - Subdural accumulation of fluid in a dog after the insertion of a ventriculoperitoneal shunt. AB - A dog with hydrocephalus as a result of aqueduct stenosis and cerebellar herniation underwent ventriculoperitoneal shunting. Magnetic resonance images and computed tomography scans taken after the surgery revealed subdural accumulations of haemorrhagic fluid and cereberocortical collapse caused by overshunting and leakage of cerebrospinal fluid from the site of insertion of the shunt. However, the degree of cerebellar herniation was reduced after the shunt was inserted, and the dog did not develop any neurological signs and made good progress. PMID- 15747659 TI - Outbreak of enteric cryptosporidiosis in cockatiels (Nymphicus hollandicus). PMID- 15747660 TI - Mycobacterium avium subspecies paratuberculosis in fallow deer and wild boar in Spain. PMID- 15747662 TI - Malignant cerebellar lymphoma in a calf. PMID- 15747661 TI - Papular-like glossitis in a dog with leishmaniosis. PMID- 15747663 TI - Progress with the RCVS practice standards scheme. PMID- 15747664 TI - Restrictions on distance retail selling of POM products. PMID- 15747665 TI - Out-of-hours cover. PMID- 15747666 TI - Regulation of paraprofessionals. PMID- 15747667 TI - Risk of tapeworm metacestode-related lesions in cattle in Britain. PMID- 15747668 TI - Clindamycin resistance in methicillin-resistant Staphylococcus aureus isolated from animals. PMID- 15747669 TI - Astroviruses found in the intestinal contents of guinea fowl suffering from enteritis. PMID- 15747670 TI - Surface analysis and effects on interfacial bone microhardness of collagen-coated titanium implants: a rabbit model. AB - PURPOSE: The aim of this study was to evaluate the surface chemistry and the microhardness at the implant-bone interface using a recently developed collagen coated titanium implant in a short-term rabbit model. MATERIALS AND METHODS: Surface chemistry was evaluated by x-ray photoelectron spectroscopy (XPS), while in vivo studies involved 4-week implants mid-diaphysis in the lateral femurs of adult male rabbits. After conventional embedding and evaluation of histologic sections, the resinembedded blocks containing the implanted screws were used to measure bone hardness by means of an indentation test. RESULTS: Decomposition of the C1s peak obtained by XPS analysis confirmed that surface-immobilized collagen retained all the molecular features of the control, nonimmobilized reference. As to microhardness measurement, newly formed bone at the collagen-coated implant/bone interface was significantly harder than bone at the interface of the uncoated control implant and bone. DISCUSSION: These results suggested that collagen coating significantly improves bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium. Surface modification of titanium implants by collagen coating has recently been discussed as a promising approach to the biochemical modification of implant surfaces. The present results support previous histologic findings and demonstrated that the biomolecular layer linked over the titanium implant can increase the bone healing rate, at least in this animal model. CONCLUSIONS: The present microhardness measurement at the bone-implant interface showed that collagen coating can significantly improve bone maturation and mineralization at the interface in comparison with uncoated commercially pure titanium, confirming and substantiating previous findings by histomorphometric measurements from the same model. PMID- 15747671 TI - The effects of ion beam-assisted deposition of hydroxyapatite on the grit-blasted surface of endosseous implants in rabbit tibiae. AB - PURPOSE: This study was undertaken to evaluate ion beam-assisted deposition (IBAD) of hydroxyapatite (HA) on the grit-blasted surface of endosseous dental implants 6 weeks postplacement. MATERIALS AND METHODS: A total of 40 implants was placed in the tibiae of 10 New Zealand white rabbits. Twenty implants were grit blasted only and the other 20 were grit-blasted and coated with HA by the IBAD method. After 6 weeks of healing, the rabbits were sacrificed and removal torque tests, histomorphometry, and morphometric analysis of microtomographic images were performed. RESULTS: The HA-coated group showed significantly higher removal torque, bone-to-implant contact, and bone volume than the other group. DISCUSSION AND CONCLUSION: In a previous study, the authors suggested that HA coating deposited on a machined surface by the IBAD method showed results comparable to or more favorable than the results obtained with a blasted surface. This study indicated that the HA coating produced by the IBAD method was also very effective on the aluminum oxide-blasted surface, as demonstrated by the early formation of osseointegration. Morphometric analysis by microtomography showed some promise in measuring the osseointegration rate. (More than 50 references.) PMID- 15747672 TI - Immediate versus delayed loading of dental implants in the maxillae of minipigs: follow-up of implant stability and implant failures. AB - PURPOSE: To assess the course of the stability and the failure rate of dental implants placed in the partially edentulous maxillae of minipigs. MATERIALS AND METHODS: Three months after tooth removal, implants were placed in 9 minipigs. Six implants (XiVE; Friadent, Mannheim, Germany) were placed on each side of the posterior maxilla after preparation of the implant sites either by an osteotome technique or with spiral drills. Implant stability was assessed by resonance frequency analysis (RFA) at the time of placement, at second-stage surgery (which took place after a healing periods of 1, 2, 3, 4, or 5 months), and after a loading period of 6 months. RESULTS: Implant stability was significantly influenced by the healing period (P = .007). Implant stability decreased after 1 to 3 months of healing for both of the placement techniques and increased after a healing period of 4 months. After implant site preparation by an osteotome technique, 6 of 12 immediately loaded implants, 18 of 24 implants loaded after healing periods of 1 to 3 months, and 1 of 18 implants loaded after a healing period of 4 or 5 months were lost. After implant site preparation using spiral drills, 7 of 12 immediately loaded implants, 12 of 24 implants loaded after healing periods of 1 to 3 months, and 2 of 18 implants loaded after healing periods of 4 or 5 months were lost. Broad overlapping of confidence intervals for the number of implant failures revealed that there was no relevant difference between immediate and early functional loading for either of the 2 techniques. DISCUSSION AND CONCLUSION: Implant loading after healing periods of 1 to 3 months did not improve implant survival compared to immediate loading in the posterior maxillae of minipigs. Not until a healing period of 4 months was reached did implant stability begin to increase. Only when functional loading was started at this point in time was maximal implant survival achieved. PMID- 15747673 TI - Mechanical and histologic examination of titanium alloy material treated by sandblasting and anodic oxidization. AB - To evaluate the biocompatibility and the bone-bonding strength of new titanium alloy materials treated by sandblasting and anodic oxidization, 3 cylindric test pieces having different surface roughnesses were manufactured and implanted into the diaphyses of the femurs of New Zealand white rabbits. Six weeks later, shear loading tests and histologic examination were carried out. Strong interfacial bonding strength and active new bone formation were confirmed in the peripheral area of the test pieces having a surface roughness (Ra = 2.7 microm and Ra = 4.7 microm). Judging from stable fixation to shear loading in bone tissue, it was concluded that group C (Ra = 2.7 microm) had the best surface condition of the 2 groups. Further detailed examination is required to demonstrate that the surface treatment used for group C (a micro rough surface on a macro rough surface structure) can enhance active bone formation and stable fixation in bone tissue. PMID- 15747674 TI - Histologic evaluation of bone-implant contact of immediately loaded transitional implants after 6 to 27 months. AB - PURPOSE: Transitional implants (TIs) were developed as a method of providing fixed provisional restorations for the implant patient who wishes to avoid removable temporary restorations during implant healing. Success of TIs depends on achieving sufficient osseointegrated bone-to-implant contact (BIC) during the provisional prosthesis phase. To date, little data are available on the degree of BIC of these implants in function. The purpose of this study was to histologically evaluate the BIC of TIs following various periods of loading. MATERIALS AND METHODS: Histologic analysis of 33 immediately loaded implants from 21 patients was performed. All TIs had turned machined surfaces and were made of commercially pure titanium (grade 1). These transitional implants were in function for an average of 10.8 months (range: 6 to 27 months). Before the definitive restoration was provided, all of the TIs were removed with trephine drills and sent for hard tissue histomorphometric analysis. RESULTS: The average percentage of BIC was 52.9% +/- 13.81% (range 25.1% to 83%). DISCUSSION: Although TIs are traditionally removed when the definitive implants are restored, the BIC and clinical integration of the TIs in the present study may suggest a change in TI protocol. Studies are indicated to examine long-term use of TIs as sole support or in conjunction with definitive implants in definitive implant supported restorations. CONCLUSION: The percentage of BIC achieved with TIs was similar to that documented in the literature for conventional turned, machine surfaced implants. PMID- 15747675 TI - Immediate loading of single-tooth implants in the posterior region. AB - PURPOSE: The aim of this study was to evaluate the clinical response and safety of immediately loaded single-tooth implants placed in the posterior region of the maxilla and mandible. MATERIALS AND METHODS: Single-tooth implants were placed in healed extraction sites in 20 adult patients. Temporary pre-fabricated acrylic resin crowns were prepared and adjusted. The crown occlusion was adjusted to obtain minimal contacts in maximum intercuspation. After 6 weeks a ceramometal or all-ceramic crown was cemented. Radiographic and clinical examinations were made at baseline and at 3, 6, and 12 months. Cortical bone response and peri-implant mucosal responses were evaluated. RESULTS: The marginal bone level at the time of implant placement was preserved. The mean change in marginal bone level was 0.01 mm at 12 months. The mean Periotest value after 360 days was -4. The peri-implant mucosal adaptation to the anatomic form of the provisional crown resulted in a natural esthetic outcome, and a gain in papilla length was observed. One implant failure was recorded because of provisional luting cement impaction. DISCUSSION: Clinical research has shown that immediate loading is a possible treatment modality. The immediate functional loading of implants placed in this study resulted in bone adaptation to loading. A satisfactory success rate with positive tissue responses was achieved. CONCLUSIONS: The results of this limited investigation indicated that immediate loading of unsplinted single-tooth implants in the posterior region may be a viable treatment option with an esthetic outcome. PMID- 15747677 TI - Success and failure rates of osseointegrated implants in function in regenerated bone for 72 to 133 months. AB - PURPOSE: Guided bone regenerative therapy has become a significant component of clinical implant practice. Initial reports have demonstrated success rates of implants in regenerated bone under function comparable to the success rates of implants placed in native nonregenerated host bone. This report documents the success and failure rates of osseointegrated implants placed in regenerated bone for up to 133 months in function. MATERIALS AND METHODS: A retrospective analysis of a group of 607 titanium plasma-sprayed cylindric implants placed in regenerated bone, the success and failure rates of which were previously reported at 6 to 51 months in function, were assessed. RESULTS: The implants demonstrated cumulative success rates of 97.2% for the maxilla and 97.4% for the mandible, yielding an overall cumulative success rate of 97.4% for up to 133 months in function. DISCUSSION AND CONCLUSIONS: Titanium plasma-sprayed osseointegrated implants of various diameters, lengths, and designs, utilized in a variety of clinical scenarios, demonstrated functional cumulative success rates comparable to those of implants placed in nonregenerated host bone for extended periods of time in this patient population. PMID- 15747676 TI - An assessment of crown-to-root ratios with short sintered porous-surfaced implants supporting prostheses in partially edentulous patients. AB - PURPOSE: Implant length, implant surface area, and crown-to-root (c/r) ratio and their relationship to crestal bone levels were analyzed in 2 groups of partially edentulous patients treated with sintered porous-surfaced dental implants. MATERIALS AND METHODS: One hundred ninety-nine implants were used to restore 74 partially edentulous patients with fixed prostheses. Implants were categorized according to their length ("short" versus "long") and estimated surface area ("small" versus "large"). "Short" implants had lengths of 5 or 7 mm, while "long" implants were either 9 or 12 mm in length. "Small" implants had estimated surface areas of < or = 600 mm2, while "large" implants had estimated surface areas > 600 mm2. Other data collected included c/r ratio (measured on articulated diagnostic casts), whether or not the implants were splinted, and standardized sequential radiographs. RESULTS: The mean c/r ratio was 1.5 (SD = 0.4; range 0.8 to 3.0), with 78.9% of the implants having a c/r ratio between 1.1 and 2.0. Neither c/r ratio nor estimated implant surface area (small or large) affected steady-state crestal bone levels. However, implant length and whether the implants were splinted did appear to affect bone levels. Long implants had greater crestal bone loss (0.2 mm more) than short implants; splinted implants showed greater crestal bone loss (0.2 mm more) than nonsplinted ones. These differences were statistically significant. DISCUSSION AND CONCLUSIONS: Sintered porous-surfaced implants performed well in short lengths (7 mm or less) in this series of partially edentulous patients. The data suggested that long implants and/or splinting can result in greater crestal bone loss; longer implants and splinted implants appeared to favor greater crestal bone loss in this investigation. These conclusions are, of course, specific to the implants used and would not be relevant to other implant types. PMID- 15747678 TI - Early loading of sandblasted, acid-etched implants in the posterior maxilla and mandible: a 1-year follow-up report from a multicenter 3-year prospective study. AB - PURPOSE: The aim of the present study was to evaluate the concept of an immediate loading protocol in the posterior maxilla and mandible through analysis of implant survival at 1 year. MATERIALS AND METHODS: One year follow-up data of a multicenter study are reported. Eighty-two ITI sandblasted, acid-etched (SLA) implants in 40 patients were loaded between 0 and 11 days after implant placement (mean 4.3 +/- 2.8 days). The restorations consisted of either 2 splinted crowns or a 3-unit fixed prosthesis. All restorations were put into full functional occlusion. Periapical radiographs were evaluated for changes in crestal bone level from baseline to 1 year postloading. Primary stability of the implants was checked initially and before the fitting of the definitive prosthesis. The restorations were evaluated by the practitioners for retention, stability, and esthetics. RESULTS: Three patients' implants were not loaded because of lack of primary stability, and a fourth patient was excluded from the study because of a protocol violation (more than 4 implants were used). All 4 patients were successfully treated outside the protocol. The overall survival rate of the remaining implants at 1 year was 98.8%. The mean bone loss at 1 year was 0.52 +/- 0.98 mm, which is within the reported limits of less than 1 mm (range 0.4 to 1.4 mm) loss in the first year. DISCUSSION AND CONCLUSION: The early results from this study indicate that early and immediate loading of 2 implants in the posterior maxilla and mandible may be suitable in selected patients. On the basis of 1 year of observation, the results appear similar to those achieved with a delayed procedure. PMID- 15747679 TI - Intraoperative computerized navigation for flapless implant surgery and immediate loading in the edentulous mandible. AB - Computerized navigation surgery has evolved to facilitate minimally invasive procedures, the gold standard of surgery today. While flapless implant surgery may be clinically beneficial, it has generally been perceived as a blind procedure limited to straightforward cases that do not pose a risk of cortical plate perforation. The objective of this report is to describe a protocol for flapless implant placement in a completely edentulous mandible using computerized navigation surgery. The Image Guided Implantology system (IGI, DenX Advanced Dental Systems) is described. The IGI system provides real-time imaging of the dental drill and transforms flapless implant surgery into a fully monitored procedure. The highly accurate intraoperative navigation enables precise transfer of the detailed presurgical implant plan to the patient. This is particularly valuable in edentulous jaws lacking any indication of the dental arch. The accurate positioning of the implants, based on the presurgical digital plan, allows fabrication of a provisional fixed prosthesis before the implant surgery for immediate postoperative loading. This innovative protocol can enhance prosthodontic-driven placement of implants in a fully monitored flapless surgery. PMID- 15747680 TI - Comparative analysis study of 702 dental implants subjected to immediate functional loading and immediate nonfunctional loading to traditional healing periods with a follow-up of up to 24 months. AB - PURPOSE: The aim of this study was to clinically evaluate immediate functionally loaded (IFL) and immediate nonfunctionally loaded (INFL) implants for various indications compared to a control group with a conventional healing period. MATERIALS AND METHODS: Two hundred fifty-three patients took part in the study. A total of 702 XiVE implants (Dentsply/Friadent, Mannheim, Germany) were placed: 253 IFL implants, 135 INFL implants, and 314 controls. RESULTS: In each of the 3 groups, 2 implants failed. For all the other implants involved, from a clinical and radiographic point of view, osseointegration was successful. DISCUSSION: As long as the prerequisites are fulfilled, immediate functional loading and immediate nonfunctional loading are predictable techniques, not only in completely edentulous patients but also in partially edentulous patients. CONCLUSION: Immediate functional loading and immediate nonfunctional loading appear to be techniques that can provide satisfactory implant success rates in selected cases. PMID- 15747681 TI - An in vitro evaluation of ZiReal abutments with hexagonal connection: in original state and following abutment preparation. AB - PURPOSE: Laboratory processing of implant-supported prostheses may alter the surface of the abutment in contact with the implant head, with potential repercussions for the interface fit. The purpose of this study was to assess changes at the implant interface of high-strength zirconia ceramic esthetic abutments with a hexagonal connection (ZiReal; 3i/Iimplant Innovations, Palm Beach Gardens, FL) following abutment preparation for single-tooth restorations. MATERIALS AND METHODS: The depth (d) and width (w) of the titanium hexagonal portion of the abutment, the apical diameter of the abutment (D), and the rotational freedom (R) of the abutment were assessed for 20 ZiReal abutments prior to preparation (time 0) and following abutment preparation (time 1) to detect any eventual change of fit of the abutment on the top of the implant hexagon. RESULTS: No significant differences relative to any study parameter (d, w, D, and R) were observed between time 0 and time 1 (P = .9542). DISCUSSION AND CONCLUSIONS: The hexagonal misfit of the titanium machined ZiReal abutment on the implant hexagon may be implicated in screw joint loosening. The results of this report suggest that if all laboratory steps are carefully observed, changes at the implant/ZiReal abutment do not occur. The maintenance of the original features of the ZiReal abutment may reduce the risk of screw loosening. PMID- 15747683 TI - Comparison of the platelet concentrate collection system with the plasma-rich-in growth-factors kit to produce platelet-rich plasma: a technical report. AB - PURPOSE: The aim of this study was to compare a new method for the production of platelet-rich plasma (PRP), the plasma-rich-in-growth-factors kit (PRGF kit; G.A.C. Medicale San Antonio, Vitoria, Spain), with an established method, the Platelet Concentrate Collection System (PCCS; 3i/Implant Innovations, Palm Beach Gardens, FL) with respect to resulting cellular and growth factor contents. MATERIALS AND METHODS: Whole blood was drawn from 51 healthy donors (20 men, 31 women) aged 19 to 59 years (mean +/- SD 35.12 +/- 9.65 years), and PRP was prepared by both methods. RESULTS: Platelet counts differed significantly (signed rank test, P < .001 for all) between the donor blood (274,200 +/- 54,050/microL), the PCCS PRP preparation (1,641,800 +/- 426,820/microL), and the PRGF kit PRP preparation (513,630 +/- 139,470/microL). The PCCS concentrated leukocytes (whole blood, 6,992 +/- 2,011/microL; PCCS PRP, 14,153 +/- 7,577/microL), while the PRGF kit produced a leukocyte-poor PRP (65 +/- 108/microL). Higher concentrations of transforming growth factor beta1 (TGF-beta1) and platelet-derived growth factor AB (PDGF-AB) were found in the PCCS PRP (TGF-/beta1, 290 +/- 95 ng/mL; PDGF-AB, 157 +/- 62 ng/mL) than in the Anitua PRGF kit PRP (TGF-beta1, 73 +/- 26 ng/mL; PDGF-AB, 47 +/- 21 ng/mL). Statistical analysis showed significant differences (P < .001 for TGF-beta1 and P < .01 for PDGF-AB). DISCUSSION: The results of this study and some data in the literature indicate that the content of growth factors in PRP can vary tremendously, depending on the system used for the preparation of PRP. CONCLUSION: PCCS collects more platelets and leukocytes than the PRGF kit. This results in significantly higher growth factor levels. Further in vivo studies are needed to determine whether this results in a clinically different biologic effect. PMID- 15747682 TI - Single preoperative dose versus long-term prophylactic antibiotic regimens in dental implant surgery. AB - PURPOSE: The purpose of this prospective study was to compare the efficacy of prophylactic antibiotic regimens commonly used in dental implant surgery. Preoperative single-dose and long-term prophylactic antibiotic regimens were compared. MATERIALS AND METHODS: This was a 2-center prospective study in which 215 patients underwent ambulatory endosseous implant placement. In the first group, 445 dental implants were placed in 125 patients after the administration of a single preoperative dose of prophylactic antibiotic with no postoperative antibiotics. In the second group, 302 dental implants were placed in 90 patients who received a preoperative dose of antibiotics and were instructed to take antibiotics postoperatively for 7 days. In both groups, 0.12% chlorhexidine pre- and postoperative mouth rinses were used. Patients returned for postoperative evaluation at 1 week, 2 weeks, and just prior to surgical uncovering. The surgical sites were assessed for pain, swelling, erythema, and purulence. RESULTS: In the first group, 3 patients developed wound dehiscence at 5 implant sites and 1 developed a minor inflammatory response. None of the patients in this group received further antibiotics. In the second group, 3 patients developed wound dehiscence, 2 developed an inflammatory response, and 1 was diagnosed with infection, for which another course of antibiotics was required. There was no statistical difference between the 2 groups according to the Fisher 2 x 4 exact test (P = .56). DISCUSSION: Indiscriminate use of antibiotics is unacceptable in clinical practice today. Surgeons must adhere to basic principles to gain the most benefit from the use of prophylactic antibiotics. CONCLUSION: Long-term prophylactic antibiotic use in implant surgery was of no advantage or benefit over a single-dose preoperative antibiotic regimen in this patient population. PMID- 15747684 TI - Human histologic and histomorphometric analyses of hydroxyapatite-coated implants after 10 years of function: a case report. AB - PURPOSE: No consensus exists on the long-term performance of hydroxyapatite (HA) coatings on dental implants. The aim of this study was to evaluate the long-term stability of the HA coating in a human autopsy specimen. MATERIALS AND METHODS: Two mandibular HA-coated implants were retrieved postmortem from a woman after 10 years of functional loading with an implant/tooth-supported fixed partial denture. After ground sectioning, the specimens were histomorphometrically analyzed. RESULTS: Direct bone-implant contact was found at 78.48% of the implant surface. HA coating disappearance had occurred in a few areas (22.75%), but bone was in direct apposition to the titanium surface. Bone volume measured 27.66%, and expected bone-implant contact was 37.55%. No inflammatory reaction was seen in the supracrestal soft tissues or the bone compartment. DISCUSSION: Most of the HA coating was maintained on the implants, and areas lacking HA were directly apposed by bone. This observation suggests that the underlying titanium surface should have a macro-texture to promote the adaptation of bone to the titanium surface in case of HA disappearance, as well as to decrease failure at the HA titanium interface. CONCLUSIONS: In a patient in whom prosthetic treatment was appropriately performed and proper plaque control was maintained, the HA coating was not damaged and contributed to the success of the implant over 10 years of clinical functioning. PMID- 15747686 TI - Comparison of classical and derivative UV-spectrophotometric methods for the quantification of diltiazem and mexiletine. AB - A first order derivative UV-spectrophotometric method for the determination of diltiazem hydrochloride and mexiletine hydrochloride has been developed and validated. In the assay, the first- and second-order measurements with the use of the "peak-zero" and "peak-peak" techniques were applied. The linear correlation (r < 0.9999) between the amplitude of the peak and the concentration of the examined drugs in the range of 3.0-8.0 microg mL(-1) for diltiazem and 50-100 microg mL(-1) for mexiletine was obtained. The proposed method was successfully applied for accurate (mean recovery about 100%), precise (RSD about 1%) and selective determination of the studied drug in the pure and dosage forms. PMID- 15747685 TI - Localized vertical maxillary ridge preservation using bone cores and a rotated palatal flap. AB - The aim of vertical augmentation of the alveolar ridge is to restore resorbed alveolar ridges. This technique is critical to the placement of dental implants in a favorable position and the enhancement of restoration esthetics. The present report describes a technique for surgical preservation of the anterior maxillary process using maxillary bone from the surgical site and raising a soft tissue rotated palatal flap. Maxillary lateral incisor extraction and periapical surgery of the central incisors were first carried out. Two bone cores were harvested from the neighboring buccal vestibular region and placed in the sockets of the lateral maxillary incisors. After 3 months, implants were placed; 12 weeks later, the prosthetic restorations were cemented. There were no complications after 2 years of follow-up. This technique constitutes a viable approach for preserving the anterior sector alveolar ridge with the posterior placement of dental implants. PMID- 15747687 TI - Esters of cephalosporins part IX. New method of isolation of cefetamet pivoxil. AB - A new method of isolation of cefetamet pivoxil, which results in higher purity and higher yield than according to the known methods, was worked out. The new method depends on ability to the adsorption on acidic active carbon of total amount of this antibiotic together with only a small amount of impurities. During two-step elution all impurities were washed out in the first step and in the next step pure cefetamet pivoxil was eluated by another solvent. PMID- 15747688 TI - Estimation of the applicability of differential spectroscopic method for the determination of lisinopril in tablets and for the evaluation of its stability. AB - Classic spectroscopy and first-, second-, third derivative UV-spectroscopy methods were applied to the determination of lisinopril (LIS) in the model solutions, tablets as well as to estimate its stability in solid phase. These results were compared with results received by the HPLC method. Derivative UV spectroscopy and the HPLC method was tested for: selectivity, precision, linearity, accuracy and repeatability. The study shows that derivative UV spectroscopy (only first and second derivative) and HPLC can be successfully applied for the quantitative analysis of lisinopril in pharmaceutical formulations and evaluation of its stability in solid phase. PMID- 15747689 TI - Topical delivery of celecoxib using microemulsion. AB - The topical delivery of celecoxib has been studied using microemulsion as the vehicle for the treatment of UV B induced skin cancer. Pseudotemary phase diagrams were constructed at different oil to cosurfactant ratios to identify the formulation variables for microemulsion formation, and the effect of these variables on skin permeation of celecoxib was evaluated with excised rat skin. Topical anti-inflammatory effect of celecoxib has been assessed using the arachidonic acid induced ear oedema model. Formulation E consisting of 3% celecoxib, 22% propylene glycol dicaprylate/dicaprate + caprylic/capric mono-/di glycerides (2:1), 30% polysorbate 80 and water (all w/w) showed higher permeation rate and significant anti-inflammatory activity. The studied microemulsion formulations have a prospect for use as a potential vehicle for treatment of UV B induced skin cancer. PMID- 15747690 TI - Prolonged delivery of ciprofloxacin hydrochloride from hydrophilic ocular inserts. AB - Ocular inserts were developed with prolonged release of drug and minimum swelling within cul-de-sac using ciprofloxacin (CPF) hydrochloride as a model drug. The ocular inserts were fahricated with sodium alginate films loaded with drug and then treated with calcium chloride. A 4% w/v solution of calcium chloride and an exposure of 15 s to this solution was found to be the optimum treatment combination of inserts. Four types of inserts were produced: type-I contained CPF hydrochloride and alginate, type-II contained CPF crystals and alginate, type-III contained CPF hydrochloride inalginate and hydroxypropylmethylcellulose (HPMC) matrix and type-IV contained CPF crystals entrapped inalginate and HPMC matrix. In vitro release profile of drug from the inserts followed Higuchi and first order kinetic models. Longer duration for 90% drug release were obtained from types-II and IV inserts than from types I and III, although type III had a longer duration than type-I inserts. In vivo studies were carried out in rabbit eyes by measuring the tear concentrations against time. From the pharmacokinctic parameters obtained types II and IV were found to prolong the duration of action more than 2 days while types I and II inserts the duration of action lasted for about 1.5 days. PMID- 15747691 TI - Design and characterization of mucoadhesive buccal patches of salbutamol sulphate. AB - Mucoadhesive patches for delivery of salbutamol sulphate were prepared using polyvinyl alcohol, hydroxypropylmethyl cellulose and chitosan. Mechanical property, swelling and bioadhesive characteristics were detemined for both plain and medicated patches. Mechanical properties were determined in presence of carbopol and polyvinylpyrrolidone. The results showed an increase in swelling after addition of salbutamol sulphate to the plain formulation. This was attributed that the salbutamol sulphate modifies the way water is bound to or taken by the polymer. A decrease in residual time was observed for polyvinyl alcohol and citosan containing formula. High drug release was obtained from polyvinyl alcohol compared to the hydroxypropylmethylcellulose. Physical characteristics of the studied patches showed promising with good bioadhesion. PMID- 15747692 TI - Studies on the syntheses of 1-substituted benzimidazole derivatives. AB - Several novel 1-heteroarylalkyl substituted benzimidazoles were synthesized in the reaction of various halogenoalkylheterocycles with benzimidazoles. The structures of the products were confirmed by their spectral and analytical data. The potential biological properties of the new compounds were evaluated using the PASS programme. PMID- 15747693 TI - Research on the connections of isoindole derivatives and ethyl esters of formic, acetic and propionic acids. AB - A series of new derivatives of isoindole in the reaction of nucleophilic substitution with ethyl chloroformate, ethyl bromoacetate and ethyl 3 bromopropanoate were synthesized. PMID- 15747694 TI - Synthesis of new derivatives of 8-aryl-3-phenyl-6,7-dihydro-4H-imidazo[2, 1 c][1,2,4]triazin-4-one. AB - Cyclocondensation of 1-aryl-2-hydrazono-imidazolidines with ethyl phenylglyoxylate furnished a novel derivatives of imidazo[2,1-c][1,2,4]triazine. PMID- 15747695 TI - The effect of acetylshikonin isolated from Lithospermum canescens roots on tumor induced cutaneous angiogenesis. AB - This study has demonstrated that acetylshikonin (ACS), the isolated ingredient from Lithospermum canescens Lehm. roots, in a daily dose of 200 microg for 3 days, inhibited cutaneous angiogenesis induced by L-1 sarcoma cells in Balb/c mice. PMID- 15747696 TI - Effect of administration of retinol and etidronate on bone histomorphometric parameters in ovariectomized rats. AB - Retinol belongs to factors affecting bone remodeling. The effect of retinol on the osseous tissue depends on the dose and duration of treatment. Retinol can cause bone damage and deformation. Retinol is frequently administered chronically in too high doses, sometimes by osteoporotic patients. The aim of the present study was to examine the interaction between retinol and an antiresorptive drug- disodium etidronate in bilaterally ovariectomized rats. The experiments were carried out on Wistar rats (200 +/- 30 g), divided into 7 groups: I--sham operated control rats. II--ovariectomized control rats (OVX), III--OVX + editronate (10 mg/kg p.o.), IV--OVX + retinol (700 IU/kg p.o.). V--OVX + retinol (3500) IU/kg p.o.), VI--OVX + etidronate (10 mg/kg p.o.) + retinol (700 IU/kg p.o.), VII--OVX + etidronate (10 mg/kg p.o.) + retinol (3500 IU/kg p.o.). The drugs were administered for 4 weeks. Bone macrometric and histomorphometric parameters of the tibia (transverse growth, width of periosteal and endosteal osteoid, area of the transverse cross-section of the diaphysis and area of the transverse cross-section of the marrow cavity) and the femur (width of epiphyseal and metaphyseal trabeculae, width of epiphyseal cartilage) were examined. Editronate partially counteracted the development of changes induced by ovariectomy. Retinol (700 IU/kg p.o.) caused decreases in the area of the transverse cross-section of the marrow cavity and the width of osteoid, and an increase in the width of trabeculae. Retinol (3500 IU/kg p.o.) caused decreases in bone mass and the area of the transverse corss-section of the marrow cavity, and an increase in the width of trabecula. Concurrent administration of etidronate and retinol in ovariectomized rats seemed not to affect bone histomorphometric parameters in a way suggesting any interaction between them. PMID- 15747697 TI - The effect of gabapentin on antinociceptive action of analgesics. AB - The effect of gabapentin at a dose of 10 mg/kg (single administration) and at 3 mg/kg/day (administered for 10 days) on the analgesic action of morphine (10 mg/kg), metamizol (500 mg/kg) and indomethacin (10 mg/kg and 1.4 mg/kg) in mice was assessed on the basis of hot-plate and tail-flick tests. All the drugs were administered intraperitoneally (i.p.). Gabapentin was administered to mice 30 min before the administration of analgesics. The animals reactions to noxious stimuli were measured 60, 90 and 120 min after gabapentin administration. Gabapentin administered in a single dose, as well as in multiple ones, was found to cause antinociception, especially evident in the hot-plate test. The single dose of gabapentin enhances the analgesic effect of morphine in the hot-plate test, whereas in the tail-flick test it demonstrates an opposite effect. Gabapentin in a single dose does not affect significantly the effects of metamizol and indomethacin, whereas multiple doses decrease the action of these drugs. Gabapentin abolishes the tolerance of antinociceptive effect of morphine. PMID- 15747698 TI - The effect of am 251, a cannabinoid CB1 receptor antagonist, on food intake in rats. AB - The anorectic effect of AM 251 (N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-di chlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide), a CB1 receptor antagonist, was studied in rats. AM 251 (0.5-2.0 mg/kg i.p.) significantly and dose-dependently reduced food intake in both free-feeding and food-deprived rats. The obtained results support the anorectic activity of CB1 receptors antagonists. PMID- 15747699 TI - [Noninvasive ventilaton for acute respiratory insufficiency]. PMID- 15747700 TI - [Popliteal fossa sciatic nerve block for ambulatory hallux valgus surgery: comparison of lateral and posterior approaches]. AB - OBJECTIVE: The aim of this study was to assess the efficacy, effectiveness, technical difficulty, and clinical usefulness of the popliteal sciatic peripheral nerve block for unilateral hallus valgux surgery and to compare the posterior and lateral approaches. We also aimed to investigate patient comfort during the procedure. MATERIALS AND METHODS: This prospective, randomized trial enrolled 60 patients scheduled for unilateral hallux valgus surgery. The patients were randomly assigned to groups of 30 patients to receive a nerve block by either a posterior or lateral approach. A nerve stimulator was used to locate the response of the two branches forming the sciatic nerve--the posterior tibial nerve and the common peroneal nerve. Then, 20 mL of 0.5% ropivacaine was injected for each nerve. Variables analyzed were efficacy, complications, quality and duration of postoperative analgesia, degree of comfort while the technique was being performed, and level of satisfaction. RESULTS: The level of comfort was good for 19 patients (70%) in the posterior approach group and for 29 (97%) in the lateral approach group (P<0.05). Level of satisfaction was good or very good for 93% and 96% of the patients in the posterior and lateral approach groups, respectively. Block efficacy was excellent for 48 patients (80%), good for 9 (15%), and poor for 3 (5%), with no significant differences between the groups. CONCLUSION: Both the posterior and lateral approaches provide easy access to the sciatic nerve for performing a block. The rate of success is high and postoperative analgesia is good, with no noteworthy complications. However, the lateral approach is more comfortable for the patient. PMID- 15747701 TI - [Comparison of 4 anesthetic techniques for in vitro fertilization]. AB - Little is known of the effect on oocytes of anesthesia administered during retrieval. The main objective of this study was to compare the prolactin and stress hormone responses to surgical stimulus under 4 anesthetic techniques and to determine whether there is an effect of technique on the number and quality of oocytes retrieved. METHODS: We designed a clinical trial to determine the plasma and follicular levels of prolactin and cortisol in patients in an assisted reproduction program. The patients were randomized to 3 anesthetic groups: general anesthesia, spinal anesthesia, or sedation with alfentanil and midazolam plus paracervical block. Patients were consecutively assigned to the fourth group to receive sedation with remifentanil plus paracervical block. RESULTS: We studied 90 patients. The patients receiving general anesthesia had the greatest increase in prolactin by the end of the procedure. Follicular cortisol increased in the paracervical block group in which remifentanil was used for sedation. The only significant difference between groups was seen for the rate of gestation of 0% in the group receiving sedation with alfentanil and midazolam before a paracervical block. Adverse effects were few with all the techniques. All patients reported a high degree of satisfaction. CONCLUSIONS: Plasma increases in prolactin and hormonal responses to follicular puncture were fully attenuated by spinal anesthesia and partially attenuated by the techniques requiring sedation. None of the anesthetic techniques proved harmful to oocytes or embryos. Nor was the effectiveness of the in vitro fertilization technique affected by any of the anesthetic techniques studied. PMID- 15747702 TI - [Initial experience with Arndt-type bronchial blockers in thoracic surgery]. AB - OBJECTIVES: The aim of the present study was to consider the feasibility of Arndt type bronchial blockers in thoracic surgery. PATIENTS AND METHODS: We enrolled the first 11 patients undergoing uncomplicated thoracic surgery expected to be of short duration. The following variables were recorded: age, weight, height, diagnosis, type of surgery, side where the blocker was used, time needed for positioning, the number of times the blocker was malpositioned according to fiberoptic bronchoscopy, time of lung isolation, duration of surgery, and effectiveness of lung collapse. RESULTS: Eleven blockers were placed in 5 women and 6 men aged between 25 and 81 years old (mean +/- SD, 52.73 +/- 22.12 years). Weight ranged from 45 to 88 Kg (mean 65.82 +/- 11.86 Kg). Height ranged from 157 to 175 cm (mean 166 +/- 5.76 cm). Time needed for positioning the blocker ranged from 2 to 30 minutes (median 5 minutes, interquartile range 1-12). According to fiberoptic bronchoscopy, the bronchial cuff was malpositioned, with herniation into the carina, in 1 case. Duration of surgery ranged from 30 minutes to 4 hours (median 70 minutes, interquartile range 50-210 minutes). Lung collapse was excellent in 9 cases and acceptable in 2 according to the surgeon. CONCLUSIONS: Our initial experience with bronchial blockers in elective surgery has shown that time for initial positioning is acceptably short and that surgical conditions are excellent in most cases. PMID- 15747703 TI - [Anesthesia for scoliosis surgery: preoperative assessment and risk screening of patients undergoing surgery to correct spinal deformity]. AB - Spinal column deformity is associated with potentially serious alterations of respiratory and cardiac function. Such alterations, in association with the underlying disease that caused the deformity--such as neuromuscular disease--are risk factors that affect the prognosis for scoliosis patients undergoing anesthesia and surgery. It is important for the anesthesiologist to carry out a detailed preoperative assessment to identify patients at risk with the aim of decreasing morbidity and mortality related to surgical correction of deformities. Of paramount importance is awareness of the risk of injury to the spinal cord that will affect function. Other issues are how to manage the patient in anomalous positions, the risk associated with hypothermia secondary to exposure of a large surgical field for a long period, and bleeding, which can sometimes exceed the patient's total volume. In the case of scoliosis surgery, all those situations may converge, obliging us to provide careful intraoperative monitoring, particularly of spinal function; to choose the appropriate anesthetic technique; and to maintain vigilance during the early postoperative period to foresee possible complications. Because scoliosis surgery is multidisciplinary, several teams must work together to assure success. PMID- 15747704 TI - [Infectious complications after epidural analgesia to control pain after abdominal surgery]. AB - Epidural abscess as a complication of spinal analgesia is rare, but because of its potential clinical implications it must always be watched for in patients who have undergone epidural procedures. The case we report is of interest because of the long latency period between removal of the catheter and the onset of clinical signs and because of the rarity of the pathogen implicated in forming the abscess (Candida albicans). PMID- 15747705 TI - [Intraoperative transesophageal echocardiography to guide removal of a hypernephroma with vena cava and right atrial extension]. AB - We describe the case of a 70-year-old man who underwent resection of a right hypernephroma with vena caval and right atrial extension with continuous monitoring by transesophageal echocardiography. The monitoring technique brought important advantages, providing images that facilitated anatomical and functional evaluation of cardiac structures without obstructing the surgical field. Images of the heart and great vessels obtained in real time allowed the surgeon to make operative decisions based on accurate assessment of the cephalad extension of the tumor to the right chambers. Transesophageal echocardiography also allowed the anesthesiologist to monitor hemodynamic status throughout surgery and watch for tumoral emboli. The imaging technique proved to be highly useful for both anesthetic and surgical management of the patient. PMID- 15747707 TI - [Intubating laryngeal mask used for difficult airway management: 3 case reports]. PMID- 15747706 TI - [Stickler syndrome: anesthetic considerations]. PMID- 15747708 TI - [Epidural anesthesia after posterior lumbar plexus block]. PMID- 15747709 TI - [Fastrach mask to resolve a difficult airway during emergency cesarean section]. PMID- 15747710 TI - [Differential diagnosis of Horner syndrome in epidural analgesia for childbirth: when should we be concerned?]. PMID- 15747711 TI - [Combined use of the Fastrach mask and fiberoptic bronchoscope in patients with full stomachs]. PMID- 15747712 TI - [Syringomyelia discovered after epidural infusion]. PMID- 15747713 TI - [Intraoperative change of anesthetic technique in a patient with intermediate Werdning Hoffman (type II) disease]. PMID- 15747714 TI - [Large vessel complication during lumbar disk surgery]. PMID- 15747715 TI - [Iatrogenic rupture of the trachea]. PMID- 15747716 TI - [The effect of AM3 in the resolution of otitis media with effusion (OME) in paediatric patients]. AB - OBJECTIVE: Otitis media with effusion (OME) is one of the most frequent diseases and the main cause of hearing loss in childhood. The medical treatment for OME includes antibiotics, antihistamine-decongestant combinations, steroids and Eustachian-tube autoinflation as the Valsalva manoeuvre. METHODS: A total of 62 children aged 2 to 8 years with a diagnosis of OME were enrolled in a prospective study to assess the resolution of OME. Patients were given antibiotics treatment with or without associated AM3. RESULTS: After 2 months of treatment, the rate of complete resolution of OME was greater in AM3 treated group than in the non-AM3 treated group (57.6% vs 27.6%) with a statistically significant difference (p<0.017). CONCLUSIONS: These results show that AM3 treatment has a beneficial effect on OME resolution when it is associated to antibiotic treatment. PMID- 15747717 TI - [Superior semicircular canal dehiscence syndrome. Embryological and surgical consideration]. AB - OBJECTIVE: Presenting the first case of superior semicircular canal dehiscence syndrome in the Spanish literature and to establish, using embryological studies, the period in wich superior semicircular canal dehiscence originates. MATERIAL AND METHODS: 52 embryos and foetuses, from 6 mm to foetal maturity, were studied. The case of a patient suffering from superior semicircular canal dehiscence syndrome is presented. RESULTS: The superior semicircular canal and the intracranial space are communicated through bony lacunae, in the period between the 24th and 28th week of foetal development, but this communication is discontinued in the 30th week. Permeability of these lacunae, later in life, could result in the development of superior semicircular canal dehiscence syndrome. The clinical results of the surgical repair of this abnormal communication, in this particular case, using a middle fossa approach and a transmastoid approach is presented. CONCLUSIONS: Superior Semicircular Canal Dehiscence Syndrome could be due to an abnormality in foetal development and its genesis, therefore, could be prenatal. Surgical repair via transmastoid approach is a reasonable alternative treatment to the middle fossa approach. PMID- 15747718 TI - [Electrooculography. Its value in the diagnosis of the patient with a balance disorder]. AB - INTRODUCTION: Controversy persists on the value of electrooculography (EOG) in the diagnosis of the unbalanced patient. The aim of this study has been to know the utility of EOG in the diagnosis of patients with equilibrium disorders. MATERIAL AND METHODS: We have examined 1000 patients in whom EOG test has been performed for unbalance symptoms. Results have been classified in peripheric or central pattern. Those patients included in the central pattern group have been compared with the results of imaging techniques. RESULTS: 45.7% of EOG performed showed pathological signs, 29.2% were of peripheral characteristics and 16.5% of central ones, of whom 6% showed different pathologies in the imaging test. DISCUSSIONS AND CONCLUSIONS: EOG in now a days of high value in the diagnosis of unbalanced patient, specially in those cases in which other clinical explorations were normal. PMID- 15747719 TI - [Our findings in the sleep endoscopy exams]. AB - INTRODUCTION: Sleep endoscopy is a technique that allows a direct visualization of the pharynx during drug-induced sleep. It allows an assessment of the anatomical site of obstruction or vibration in habitual snorers. MATERIALS AND METHODS: A total of 51 patients underwent sleep endoscopy. All answered a standard questionnaire and underwent an otolaryngological exam as a part of the diagnostic procedure. RESULTS: The most frequent site of obstruction was the palate in 90.4%, followed by the base of the tongue in 38.5%. Obstruction at multiple levels was found in 60.7% of patients. No correlations were found between the level of the obstruction found in the ENT examination and the one found during the sleep endoscopy. CONCLUSIONS: Sleep endoscopy is useful for the assessment of the usual snorer and we recomend it to label the patient's obstruction. PMID- 15747720 TI - [Cidofovir activity in infantile recurrent respiratory papillomatosis]. AB - We report the case of a 4 year old boy that arrived to our department refering a 4 months history of dysphony complicated in the last week with inspiratory estridor. He was diagnosed of respiratory papillomatosis. In the next 10 months he underwent 6 operations to remove papillomas with CO2 laser. In the last operation we decided to add adjuvant therapy with intralesional injections of cidofovir once all papillomas had been removed. After more than a year of monitoring he remains disease free. PMID- 15747721 TI - [Value of nutritional support in patients with pharingocutaneous fistula]. AB - INTRODUCTION: A prospective and statistic study is presented to evaluate the efficacy of nutritional support in the postoperative care of patients with pharyngocutaneous fistula after laryngectomy. PATIENTS AND METHODS: All patients who developed pharyngocutaneous fistula after total o partial laryngectomy between 2001 and 2004 were included and assigned to the study group if a supplementary and enteral nutrition was started through the nasogastric tube, and to the control group if a conventional liquid diet was given. Nutrition of patients in the study group was controlled by an Endocrinologist. RESULTS: 32 patients were included (14 in the study group and 18 in the control group). Both groups were similar for all variables measured except for the length of hospital stay (mean difference 31.02 days) and the need for surgical closure (33% in the control group and 0% in the study group). These differences were found statistically significant. DISCUSSION AND CONCLUSIONS: Postoperative supplementary enteral nutrition controlled by the Endocrinologist reduced the need for surgical treatment of pharyngocutaneous fistula and shortened the stay in hospital of these patients. More studies are needed to prove the efficacy of preoperative nutrition in reducing the incidence of fistula after laryngectomy. PMID- 15747722 TI - [Multimedia educational material. A tool of support to the Otolaryngology training]. AB - Due to multimedia technologies, the medical information has taken an extraordinary growing. Nevertheless, the education at University will turn out to be a more complex pedagogic process. We present a multimedia educational material (MEM), developed as an online course to "Seminars of Otolaryngology", and available free at the University website. MEM was designed and edited by, ENT teachers, pedagogues, and image and computer experts. It has been structured as a series of modules that integrate texts, images, movie clips, video, audio and animations. The schematic presentation stands out with progressive access to the topics through internal and external links. The pedagogic resources give additional support by learning objectives, glossary, references, index and searcher. Finally, the activities and instrumental simulations complete the training in diagnosis and treatment. The MEM is an interactive useful software program for medical training in Otolaryngology, without avoiding the key figure of the teacher. PMID- 15747723 TI - [Primitive neuroectodermal tumor/extraosseus Ewing sarcoma of the temporal bone]. AB - The term primitive neuroectodermal tumor (PNET) should be included in the differential diagnosis of small round blue cell tumors, which come from the neural crest and explain their neuroectodermal character. This is one of the most indifferentiated histological types of malignant tumors. In the PNET family, we high light the Ewing's Sarcoma, that is frecuently located in bony sites and has a less frequent localization in soft tissues. We focus on one atypic case of PNET/Extraosseus Ewing's Sarcoma that involves the external auditory canal and infiltrates the temporal bone. Surgery followed by postoperative chemotherapy and radiation offers the best local control of the disease. PMID- 15747724 TI - [Endoscopic nasal surgery for pterigopalatine fossa schwannoma]. AB - Schwannomas are tumors of nerve sheath origin, the lesion being derived from the Schwann cells surrounding neural tissue in most of the peripheral, cranial and autonomic nerves. Schwannomas occurring in the head and neck represent 25-35% of all reported schwannomas, and most commonly arise from the acoustic or the neumogastric nerves. In the nose and paranasal sinuses these tumors are very uncommon (fewer than 4% of all schwannomas). In the pterygopalatine fossa they are very rare, and arise in this case from the ophthalmic and maxillary branches of the trigeminal nerve. In this exceptional site, the tumor is classically approached through a Caldwell-Luc incision. In this case, we report the removal of a schwannoma of the pterygopalatine fossa by endoscopic sinus surgery. PMID- 15747725 TI - High variability of milk protein genes in Bos indicus cattle breeds of Cameroon and Nigeria and characterization of a new alphas1-casein promoter allele. AB - The study provides the first comprehensive information on the variability of milk protein genes of Bos indicus and Bos taurus cattle breeds in Cameroon and Nigeria. The investigations indicate a high diversity of milk protein genes for the zebu populations. Of the investigated alleles, 21 out of 29 were observed. The method of single strand conformation polymorphism (SSCP) was a particularly useful technique because it allowed discrimination of alleles, including zebu specific alleles at the CSN2 (I) and CSN3 (A(I) and H) loci, not separated by protein electrophoretic techniques and also made possible the detection of a further CSN1S1 5' promoter allele (CSNIS1Prom5), which is also zebu-specific. Characterization of CSN1S1Prom5 showed that it was the most variable of all described CSN1S1 promoter alleles. A potential GATA consensus motif is created by mutations in CSN1S1Prom5. Intra-breed diversity measured as mean effective number of alleles was higher in the zebu populations than in the taurine breeds. Of the expected casein haplotypes, 96 out of 320 were present in the studied breeds. 2-C A-A2-H (CSN1SIProm2-CSN1S1C-CSN1S2A-CSN2A2-CSN3H) and 5-C-A-A2-H were zebu specific while 1-B-A-A2-B was specific to the taurines. Overall distribution of alleles and haplotypes clearly separated the zebu populations from the taurine breeds. Zebu influence on the taurine breed Namchi was detected through the occurrence of zebu alleles and haplotypes. High variability of milk proteins also means availability of resources for breed development, phylogenetic studies, and conservation and management decisions. PMID- 15747727 TI - Homofermentative lactic acid bacteria of a traditional cheese, Comlek peyniri from Cappadocia region. AB - Comlek peyniri is a typical artisanal cheese in Central Anatolia. This type of cheese was made by using the indigenous lactic acid bacteria (LAB) flora of cow or ewes' milk. Majority of the samples were taken from fresh cheese because the aim was to isolate homofermentative LAB. Initially 661 microbial isolates were obtained from 17 cheese samples. Only 107 were found to be homofermentative LAB. These isolates were selected and identified by using both phenotypic and molecular methods. Phenotypic identification included curd formation from skim milk, catalase test, Gram staining and light microscopy, growth at different temperatures and salt concentrations, arginine hydrolysis, gas production from glucose, and carbohydrate fermentation. Molecular identification was based on the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the 16S rRNA gene-ITS (internally transcribed spacer) region. By combining the phenotypic and molecular identification results, isolates belonging to each of the following genera were determined at species or subspecies level: 54 Lactococcus lactis subsp. lactis, 21 Enterococcus faecium, 3 Ec. faecalis, 2 Ec. durans, 10 Ec. sp., 15 Lactobacillus paracasei subsp. paracasei, and 2 Lb. casei strains. Technological characterisation was also performed by culturing each of the strains in UHT skim milk, and by monitoring pH change and lactic acid production at certain time intervals through the 24 h incubation. Results of the technological characterisation indicated that 33% of the isolates (35 strains) were capable of lowering the pH of UHT milk below 5.3 after 6 h incubation at 30 degrees C. Thirty four of these strains were Lc. lactis subsp. lactis, and only one was an Ec. faecium strain. PMID- 15747726 TI - Effect of different milking routines on milking-related release of the hormones oxytocin, prolactin and cortisol, and on milk yield and milking performance in Murrah buffaloes. AB - Milking-related release of oxytocin, prolactin, and cortisol was studied following three premilking treatments. Six Murrah buffaloes were treated with direct application of milking cluster (O), a 1-min pre-stimulation (M), and combined feeding and pre-stimulation (MF). Machine milk yield, stripping yield and milk composition were recorded. Milk ejection occurred significantly earlier with MF than M and O (P<0.05; 2.50, 5.10 and 6.33 min, respectively). In all treatments, milk ejection occurred with small increases >3-5 ng/l in oxytocin concentration. Increase in oxytocin concentration over a threshold level and milk ejection occurred simultaneously and were closely correlated (r=0.83, P<0.05). There was a positive correlation between total time oxytocin concentration remained elevated over threshold levels and machine yield (r=0.86, P<0.05). For treatment O, milk ejection was inhibited during machine milking, while a marked increase in oxytocin occurred during hand stripping (6 and 16 ng/l, respectively). For treatment M, mean oxytocin concentrations remained unchanged during prestimulation but increased during subsequent machine milking and hand stripping (6.38, 18.06 and 12.36 ng/l, respectively). For treatment MF, although there was a 3.6-fold increase during pre-stimulation, oxytocin increased by 10 fold and 3-fold during machine milking and hand stripping, and was significant for machine milking (P<0.05, 17.32, 47.86, 18.13 ng/l, respectively). Milk ejection-related cortisol release was visible only in treatment MF. For treatments O and M, prolactin concentration increased prior to the increase in oxytocin. The stripping yield was higher, and fat content in the stripping yield significantly lower, for treatment O indicating incomplete milking. Thus buffaloes are easily disturbed even by small changes in milking routines. PMID- 15747728 TI - Potential applications of high pressure homogenisation in processing of liquid milk. AB - Studies of the potential of high pressure homogenisation (HPH) for the combined pasteurisation/ homogenisation of raw bovine milk were undertaken. Raw milk was preheated to 45 degrees C and HPH-treated at 150, 200 or 250 MPa; milk outlet temperature at these pressures were 67, 76.8 and 83.6 degrees C, respectively, with a holding time of approximately 20 s. Raw and commercially pasteurized and homogenized (CPH) milk samples were analysed as controls. Fat globules in HPH samples were approximately half the size of those in CPH samples, although differences were not significant (P>0.05). beta-Lactoglobulin was denatured at pressures > or =150MPa, although little denaturation of alpha-lactalbumin was observed. Numbers of psychrotrophic bacteria in raw milk were reduced by 2.73 log cycles by HPH at 150 MPa and were uncountable following HPH at 200 or 250 MPa. Mesophilic bacterial counts were reduced by 1.30, 1.83 and 3.06 log cycles by HPH at 150, 200 or 250 MPa, respectively. No viable Staphylococcus aureus nor coliform cells remained in any HPH milk samples. HPH did not affect the colour of milk and HPH samples did not cream during refrigerated storage. The activities of plasmin, alkaline phosphatase and lactoperoxidase in milk were all greatly reduced by HPH. Pseudomonas fluorescens, inoculated into milk (approximately 10(6) cfu/ml), was reduced to undetectable levels by HPH at 200MPa (milk inlet temperature, approximately 10 degrees C); however, Ps. fluorescens proteinase was quite resistant to HPH under such conditions. Overall, owing to the significant increase in temperature and the possibility of varying the holding time, there may be potential applications for HPH as a novel liquid milk processing technique, combining many advantages of conventional homogenization and pasteurization of milk in a single process. PMID- 15747729 TI - Effect of protein concentration, pH, lactose content and pasteurization on thermal gelation of acid caprine whey protein concentrates. AB - The influence of pH (4.5-6.5), sodium chloride content (125-375 mM), calcium chloride content (10-30 mM), protein concentration (70-90 g/l) and lactose content on the gel hardness of goat whey protein concentrate (GWPC) in relation to the origin of the acid whey (raw or pasteurized milk) was studied using a factorial design. Gels were obtained after heat treatment (90 degrees C, 30 min). Gel hardness was measured using texture analyser. Only protein concentration and pH were found to have a statistically significant effect on the gel hardness. An increase in the protein concentration resulted in an increase in the gel hardness. GWPC containing 800g/kg protein formed gels with a hardness maximum at the pHi, whereas GWPC containing 300 g/kg protein did not form true gels. Whey from pasteurized milk formed softer gels than whey from raw milk. A high lactose content (approximately 360 g/kg) also reduced the gelation performance of GWPC. PMID- 15747730 TI - Effects of hormones and growth factors on TGF-beta1 expression in bovine mammary epithelial cells. AB - The decline of mammary epithelial cell (MEC) number during mammary gland involution in the cow is due to inhibition of proliferation and induction of apoptosis. Transforming growth factor-beta 1 (TGF-beta1) belongs to a group of intramammary auto/paracrine inhibitors of bovine MEC growth and inducers of apoptosis. However, the mechanism responsible for the regulation of TGF-beta1 expression in MEC is not known. The present study examined the effect of the hormones, growth hormone (GH), somatostatin (STS), 17-beta oestradiol (E2), progesterone (P4), as well as the growth factors, insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF), on TGF-beta1 expression in the bovine MEC lines, BME-UV1 and MAC-T. The model of apoptosis in bovine mammary gland in vitro was applied by reduction of fetal bovine serum (FBS) (from 10% to 2% or 0.5% FBS) in the cell environment to show the relationship between TGF-beta1 expression and apoptosis in bovine MEC. RT-PCR, Western blot and laser scanning cytometry (LSC) were used for analysis of TGF-beta1 transcript and protein level as well as apoptosis and cell cycle in examined MEC. In this model of apoptosis, FBS deficiency (mimicking the naturally occurring decline in the access of bioactive compounds and nutrients at the end of lactation and dry period) was associated with increased TGF-beta1 expression at the level of transcript and protein, induction of apoptosis and inhibition of cell cycle. Exogenous TGF beta1, IGF-I, EGF and GH inhibited FBS-deficiency-stimulated TGF-beta1 expression. The suppressive effect of GH was reversed when cells were maintained longer in FBS-deficient medium. In general, STS, E2 and P4 increased TGF-beta1 expression. However, this effect was dependent on hormone concentration and cell line. BME-UV1 cells were much more responsive to the peptides, GH, STS, IGF-I and EGF, whereas MAC-T cells were more responsive to the steroid sex hormones: E2 and P4. PMID- 15747731 TI - Visual appearance and CMT score of foremilk of individual quarters in relation to cell count of cows milked automatically. AB - The objectives of the study were: to evaluate the interaction between visual appearance and California mastitis test (CMT) score of the foremilk in relation to the cell count of the milk; to evaluate the consequences of sorting milk according to these criteria; and to explore whether visual appearance and CMT score of foremilk depended on the time interval between milkings. Measuring somatic cell count (SCC) in composite milk only and discarding milk above certain thresholds will not ensure that milk from all cows with visually abnormal foremilk is withheld from delivery. Low thresholds of SCC will reduce the frequency of cows with abnormal milk but increase the discarding of milk from cows with visually normal foremilk. CMT score of foremilk differentiated better between cows with high and low SCC in composite milk than visual inspection of foremilk. CMT scores of foremilk decreased with increasing interval between milkings within cow, whereas the visual appearance was independent of the interval. We propose that visual appearance of the foremilk should be kept as a criterion for sorting milk at time of milking. For test purposes, the use of visual appearance of foremilk for differentiation between normal and abnormal milk has to be done on multiple milkings. Additionally, CMT scoring of foremilk improves correct classification of normal and abnormal quarters and especially when including data from the previous milking. PMID- 15747732 TI - Development of a biosensor immunoassay for the quantification of alphas1-casein in milk. AB - An immunoassay to quantify alphas1-casein (alphas1-CN) in milk using an optical biosensor, based on surface plasmon resonance (SPR) measurement, has been developed. The assay consists of a two-step sandwich strategy, with two anti alphas1-CN antibodies directed against each extremity of the molecule. This strategy permits only intact alphas1-CN to be quantified and not its degradation products. The calibration curve was obtained using a reference milk powder with a known alphas1-CN concentration. Analysis time per sample was less than ten minutes. The antibody-coated surface could be used for more than 150 determinations. Detection limit was established at 0.87 microg/ml and the intra- and inter-assay variation coefficients were 2.86 and 5.31%, respectively. The method was applied to raw milk to quantify intact alphas1-CN, with no pretreatment of the sample. An initial analysis of 48 milk samples permitted alphas1-CN concentrations ranging from 8.8 to 12.06 mg/ml to be obtained. PMID- 15747733 TI - Protein composition, plasmin activity and cheesemaking properties of cows' milk produced at two altitudes from hay of lowland and high-alpine origins. AB - The influence of high altitude, alpine origin of the forage and roughage-only diets on milk protein content and composition, plasmin activity and cheesemaking properties was investigated. There were four treatment groups, each consisting of six dairy cows in early to mid-lactation. Two groups were fed only with hay ad libitum either at 2000 m or at 400 m a.s.l. One group, kept in the lowlands, was pair-fed to the alpine-site group and another group received a mixed diet of silages, hay and concentrates. Two hay types, harvested either at the alpine site or in the lowlands, were offered to all cows fed with hay alone, following a change-over design in three periods each of 3 weeks. In the respective third week, milk was sampled at every milking. Hay of alpine origin significantly reduced milk protein, in particular whey proteins, which is why the casein number increased. kappa-Casein proportion in total casein was reduced and its glycosylation was increased by the alpine hay. The apparent plasminogen-derived activity was reduced when alpine hay was given, but apparent plasmin activity and rennet coagulation properties of the milk were not affected by hay type. Independent of hay type, the high altitude group showed a significantly reduced milk protein content, lower glycosylation of kappa-casein and impaired rennet coagulation properties. For several of the traits, the trend was the same in the pair-fed group. There was no effect of altitude on apparent plasmin activity. Hay alone v. the mixed diet resulted initially in marked declines in milk protein content but did not impair cheesemaking properties. Thus the extensive diet without concentrates, typical of high-alpine conditions, contributed less to the overall effect of extensive alpine v. intensive lowland feeding systems than hay quality and altitude did. In conclusion, certain positive influences of the alpine sojourn of cows on cheese processing quality are overruled by the major adverse impact of lower milk protein content. PMID- 15747734 TI - Use of partial budgeting to determine the economic benefits of antibiotic treatment of chronic subclinical mastitis caused by Streptococcus uberis or Streptococcus dysgalactiae. AB - The economic effect of lactational antibiotic treatment of chronic subclinical intramammary infections due to Streptococcus uberis or Streptococcus dysgalactiae was explored by means of partial budgeting. Effects at cow level and herd level were modelled, including prevention of clinical mastitis episodes and the prevention of transmission of infections. Input variables for our deterministic model were derived from literature or based on 2002/2003 dairy prices and farming conditions in The Netherlands. Sensitivity analysis was used to examine the effect of uncertainty around input variables or changes in price estimates. On farms where pathogen transmission was prevented through proper udder health management, 3-d antibiotic treatment during lactation resulted in an average net profit of euro+11.62 over no treatment while 8-d antibiotic treatment had an average negative net result of euro-21.83. Sensitivity analysis showed that profitability depends on the probability of treatment-induced cure, pathogen transmission rates, culling rate, retention pay-off, and costs of antibiotic treatment. Three-day antibiotic treatment of chronic subclinical streptococcal mastitis is economically profitable over a range of input values for cure probabilities, transmission rates and losses due to culling. In contrast, 8-d lactational treatment is only profitable for very valuable animals, on farms where the risk of pathogen transmission is high and/or when the farmer is likely to cull a high percentage of cows with subclinical mastitis. Because bacterial flora, cow characteristics and management differ widely between farms, the economic outcome of lactational treatment of chronic subclinical streptococcal mastitis may be highly farm-dependent. PMID- 15747736 TI - Video microscopy as an alternative method for somatic cell count in milk. AB - This paper presents an alternative method to estimate somatic cell count (SCC) in cows' milk. SCC is an important indicator in the detection of inflammatory reactions within the udder in cows and Direct Optical Microscopy (DOM) is the present reference method for SCC but, owing to its dependence on human operators, it is extremely costly, time-consuming and potentially subjective. The industrial method of choice is Epifluorescence (EF), which has the potential for impressive throughput and acceptable precision, but requires huge inversions and handling of highly toxic reactives and waste. In this paper, an advantageous method that involves application of a low-cost Video Microscopy (VM) system is analysed and discussed, including a comparison between DOM and VM, and an example of application of both methods to evaluate EF counts. We conclude that VM is sufficiently precise and very cheap to implement and operate. PMID- 15747735 TI - Proteolytic patterns and plasmin activity in ewes' milk as affected by somatic cell count and stage of lactation. AB - A total of 120 milk samples were collected from Comisana ewes throughout lactation. The ewes were ranked into two somatic cell count (SCC) categories: normal milk (N Milk) with SCC lower than 5.00x 10(5)/ml and high somatic cell milk (HSC Milk) with SCC higher than 1.00 x 10(6)/ml. Milk samples were analysed in triplicate for pH, fat and protein contents, renneting parameters, and plasmin and plasminogen activities. The peptide profile due to total proteolytic activity (endogenous and exogenous enzymes) on alpha- and beta-CNs were determined using urea-PAGE on sodium caseinate (pH 8.0 and pH 5.0) incubated at 37 degrees C for 4 d after sampling. The peptide profile due to non-plasmin enzyme activities at pH 5.0 was also determined using urea-PAGE. Plasmin activity was higher in the HSC milk than in the N milk throughout the study period. A decrease in plasmin activity was observed in the N milk during mid-lactation, which was probably related to decrease in pH, and in the HSC milk during late lactation, which may be ascribed to an enhanced influx of plasmin inhibitors from the blood stream. Proteolytic patterns in Comisana ewe milk were mainly affected by plasmin activity that increased with the SCC in milk. Also non-plasmin proteolytic activity was strongly enhanced by elevated SCC and resulted in a higher degradation of alpha-casein than of beta-casein. In general, plasmin activity did not increase with the advancement of lactation and exhibited a different trend in HSC and N milk, suggesting that physiological factors did not play a key role in regulating the plasminogen-plasmin system in ewes' milk. Plasmin activity, detected with the colorimetric assay was consistent with proteolytic activity on sodium caseinate shown in urea-PAGE electrophoregram. PMID- 15747737 TI - Effects on milk yield of milking interval regularity and teat cup attachment failures with robotic milking systems. AB - A database consisting of 35291 milking records from 83 cows was built over a period of 10 months with the objectives of studying the effect of teat cup attachment failures and milking interval regularity on milk production with an automated milking system (AMS). The database collected records of lactation number, days in milk (DIM), milk production, interval between milkings (for both the entire udder and individual quarters in case of a teat cup attachment failure) and average and peak milk flows for each milking. The weekly coefficient of variation (CV) of milking intervals was used as a measure of milking regularity. DIM, milking intervals, and CV of milking intervals were divided into four categories coinciding with the four quartiles of their respective distributions. The data were analysed by analysis of variance with cow as a random effect and lactation number, DIM, the occurrence of a milking failure, and the intervals between milkings or the weekly CV of milking intervals as fixed effects. The incidence of attachment failures was 7.6% of total milkings. Milk production by quarters affected by a milking failure following the failure was numerically greater owing to the longer interval between milkings. When accounting for the effect of milking intervals, milk production by affected quarters following a milking failure was 26% lower than with regular milkings. However, the decrease in milk production by quarters affected by milking failures was more severe as DIM increased. Average and peak milk flows by quarters affected by a milking failure were lower than when milkings occurred normally. However, milk production recovered its former level within seven milkings following a milking failure. Uneven frequency (weekly CV of milking intervals >27%) decreased daily milk yield, and affected multiparous more negatively than primiparous cows. PMID- 15747738 TI - Dietary trans octadecenoic acids upregulate the liver gene encoding peroxisome proliferator-activated receptor-alpha in transition dairy cows. AB - Effects of feeding calcium salts of conjugated linoleic acid (CLA) or trans octadecenoic acids (trans 18:1) on lipid metabolism and hepatic contents of mRNA encoding carnitine palmitoyltransferase 1 (CPT1), microsomal triglyceride transfer protein (MTP) and peroxisome proliferator-activated receptor alpha (PPARalpha) were examined in 15 early post-partum Holstein cows. Dietary treatments were initiated at approximately 4 weeks prior to expected calving dates and continued for 7 weeks post partum. Treatments prepartum consisted of 1) a basal diet (Control), 2) basal diet+150 g/d of CLA mix (CLA), or 3) basal diet+150 g/d of trans 18:1 mix (TRANS). Intakes of calcium salts of CLA and trans 18:1 mixes were adjusted to 225 g/d during the 7-week postpartum treatment period. Blood samples were collected at weeks 1, 2 and 4 post partum and plasma was harvested immediately for subsequent hormone and metabolite assays. Concentrations of insulin, insulin-like growth factor-I (IGF-I), and leptin in blood did not vary among cows fed the three diets. Plasma nonesterified fatty acid (NEFA) concentrations decreased between weeks 1 and 4 of lactation and were lower in cows fed the diet supplemented with trans 18:1 than in those fed a control diet at week 2 post partum. Periparturient fat supplementation had no detectable effects on CPT1 mRNA content in the liver. Steady-state concentration of MTP mRNA in the liver was greater in the TRANS treatment group than in the control group at week 1 postpartum. Feeding trans 18:1 supplements to transition dairy cows upregulated hepatic PPARalpha mRNA content during the first month of lactation. Under the present experimental conditions, dietary CLA had minimal effects on plasma and hepatic lipid metabolite concentrations in early lactation Holstein cows. Results indicate that dietary trans fatty acids may affect liver lipid metabolism in post-partum dairy cows through alterations in PPARalpha gene expression. PMID- 15747739 TI - Chemical and rheological aspects of gel formation in the California Mastitis Test. AB - The rheological properties of the CMT gel were analysed. Data are presented to demonstrate that the gel is a non-homogenous, visco-elastic, non-Newtonian fluid with rheopectic, and rheodestructive behaviour. The fundamental chemistry of the CMT is reviewed and a modified theory of gel formation is presented. The implications of the rheological properties and modified theory of gel formation for an automatic sensor are discussed. PMID- 15747740 TI - Arsenic concentration in water and bovine milk in Cordoba, Argentina. Preliminary results. AB - The Chaco Pampean Plain of central Argentina constitutes one of the largest regions of high arsenic (As) groundwaters known, covering around 1 x 10(6) km2 (Smedley & Kinniburg, 2002; Farias et al. 2004). The high-As groundwaters are from Quaternary deposits of loess (mainly silt) with intermixed rhyolitic or dacitic volcanic ash (Nicolli et al. 1989, Smedley et al. 1998,2002). Early in the last century an endemic disease due to contamination of drinking water with arsenic was recognised. This disease is called HACRE (Hidroarsenicismo Cronico Regional Endemico, Chronic Endemic Regional Hydroarsenism) and is connected with a particular type of skin cancer (Astolfi et al. 1981). One of the most affected region is the province of Cordoba, where Nicolli et al. (1989) reported As concentrations that exceed the maximun level permitted for drinking water of 50 microg/l for 82% of the groundwater samples (n=60) of a study area comprising approximately 10000 km2. The southeast of Cordoba is an important milk production zone in Argentina, where dairy product consumption is up to 192 equivalent milk l/inhabitant/year. As a secretion of the mammary gland, milk can carry numerous xenobiotic substances, which constitute a technological risk factor for dairy products and above all for the health of the consumer (Licata et al. 2004). Nevertheless no studies on the incidence of high-As livestock drinking water in livestock health and its transfer to milk have been performed in Argentina. The aim of the present study was the determination of arsenic content in livestock drinking water and milk from dairy farms located in an area of high-As groundwaters, to analyse the relation between As uptake through water and its transfer to milk. PMID- 15747741 TI - Evaluation of different preservation techniques on the storage potential of Kefir grains. AB - Kefir is an acidic, mildly alcoholic dairy beverage produced by the fermentation of milk with a grain-like starter culture (Koroleva, 1988). These grains usually contain a relatively stable and specific balance of microbes that exist in a complex symbiotic relationship (Obermann & Libudzisz, 1998; Witthuhn et al. 2004). The different groups of microbes present in the grains are active at different stages of the fermentation (Koroleva, 1982). The lactococci, including Lactococcus lactis subsp. lactis, Lc. lactis subsp. cremoris and Lc. lactis subsp. diacetilactis provide rapid acid development during the first hours of the fermentation (Litopoulou-Tzanetaki & Tzanetakis, 2000). As the acidity of the milk increases it provides favourable conditions for the growth of the lactobacilli (Rea et al. 1996). The yeasts, acetic acid bacteria and the aroma producing microbes, mainly leuconostocs, have a much slower growth rate than the lactic acid producers, resulting in the slow production of the aroma compounds and the gradual increase in the concentration of these substances in the later stages of the fermentation (Koroleva, 1982). In the past the preservation of the microbial populations present in the traditional Kefir grains was achieved by methods including freezing (Garrote et al. 1997), lyophilisation (Oberman & Libudzisz, 1998), air-drying (Kroger, 1993) and refrigeration (Marshall, 1993). Research has shown that traditional Kefir grains preserved by air-drying and lyophilisation retain their activity for up to 12-18 months (Oberman & Libudzisz, 1998). Frozen grains stored at -20 degrees C were found to maintain the microbial activity for up to 7-8 months, whereas grains stored at refrigerated temperatures showed a decreased activity after about 10 d (Oberman & Libudzisz, 1998). The aim of this study was to evaluate the impact of four different preservation techniques on the activity of mass cultured Kefir grains (Schoevers & Britz, 2003). The activity of the grains was evaluated at different time intervals using four activity measurements, including changes in substrate pH, titratable acidity (TA), lactose and lactic acid levels of the final Kefir beverage. PMID- 15747742 TI - Members. American Ophthalmological Society. PMID- 15747743 TI - Chemoreduction for retinoblastoma: analysis of tumor control and risks for recurrence in 457 tumors. AB - PURPOSE: To evaluate individual tumor control following chemoreduction for retinoblastoma. METHODS: Prospective nonrandomized single-center case series of 457 retinoblastomas managed with six cycles of chemoreduction (vincristine, etoposide, and carboplatin). The tumors were then managed with chemoreduction alone (group A) or chemoreduction combined with thermotherapy (group B), cryotherapy (group C), or both thermotherapy and cryotherapy (group D). The main outcome measure was development of tumor recurrence. RESULTS: Of 457 retinoblastomas, 63 (14%) were in group A, 256 (56%) in group B, 127 (28%) in group C, and 11 (2%) in group D. The tumor was located in the macula in 33 (52%) of group A, 109 (43%) of group B, 3 (2%) of group C, and 1 (9%) of group D. Using Kaplan-Meier analysis, recurrence of the individual retinoblastoma at 7 years was found in 45% of group A and in 18% of combined groups B, C, and D. Treatment of the 93 tumor recurrences included thermotherapy, cryotherapy, or plaque radiotherapy in 62 cases (67%) and external beam radiotherapy or enucleation in 31 cases (33%). Risk factors predictive of tumor recurrence by multivariate analysis included macular tumor location for all groups and, additionally, female sex for group A and increasing tumor thickness for groups B, C, and D. CONCLUSIONS: Chemoreduction alone or combined with cryotherapy and/or thermotherapy is effective for treatment of retinoblastoma, but tumor recurrence is greatest for those located in the macula and those with greater thickness. Globe salvage is usually achieved despite tumor recurrence. PMID- 15747744 TI - The utilization of eye care services by persons with glaucoma in rural south India. AB - PURPOSE: To determine utilization of eye care services, in particular those relating to glaucoma, in a rural population of southern India aged 40 years or older. METHODS: A total of 5,150 subjects aged 40 years or older selected through a random cluster sampling technique from three districts in southern India underwent detailed ocular examinations for vision impairment, blindness, and ocular morbidity. Information regarding previous use of eye care services was collected from this population through a questionnaire administered by trained social workers prior to ocular examinations. RESULTS: One thousand eight hundred and twenty-seven persons (35.5%) gave a history of prior eye examinations, primarily from a general hospital (n = 1,073, 58.7%). Increasing age and education were associated with increased utilization of eye care services. Among the 3,323 persons who had never sought eye care, 912 (27.4%) had felt the need to have an eye examination but did not do so. Only one third of persons with vision impairment, cataracts, refractive errors, and glaucoma had previously utilized services. Of the 64 subjects diagnosed as having primary open-angle glaucoma, 32 (50%) had previously seen an ophthalmologist, but none had had an eye examination within 1 year before the study. Only six (19%) of the 32 had been diagnosed as having glaucoma (9% of all subjects found to have glaucoma in the survey). Thirteen (20.3%) of the 64 subjects were blind in either eye due to glaucoma, including one person who was bilaterally blind. CONCLUSIONS: A large proportion of persons in a rural population of southern India who require eye care are currently not utilizing existing eye care services. Strategies to improve the uptake of services are required to reduce the burden of blindness due to glaucoma in southern India. PMID- 15747745 TI - Corneal endothelium and postoperative outcomes 15 years after penetrating keratoplasty. AB - PURPOSE: To determine changes in the central endothelium and thickness of grafted corneas, and the cumulative probability of developing glaucoma, graft rejection, and graft failure 15 years after penetrating keratoplasty. METHODS: In a longitudinal cohort study of 500 consecutive penetrating keratoplasties by one surgeon, regrafted eyes, fellow eyes of bilateral cases, and patients not granting research authorization were excluded, leaving 388 grafts for analysis. At intervals after surgery, we photographed the endothelium and measured corneal thickness by using specular microscopy. The presence of glaucoma, graft rejection, and graft failure was recorded. RESULTS: The 67 patients examined at 15 years represented 30% of the available clear grafts (107 patients had died, 76 grafts had failed). Endothelial cell loss from preoperative donor levels was 71 +/- 12% (mean +/- SD, n = 67), endothelial cell density was 872 +/- 348 cells/mm2, and corneal thickness was 0.59 +/- 0.06 mm. Endothelial cell density was unchanged between 10 and 15 years (minimum detectable difference was 96 cells/mm2, alpha = .05, beta = .20, n = 54), whereas corneal thickness increased (P = .001, n = 55). The mean annual rate of endothelial cell loss from 10 to 15 years after surgery was 0.2 +/- 5.7% (n = 54). The cumulative probability of developing glaucoma, graft rejection, or graft failure was 20%, 23%, and 28%, respectively, and six of the eight graft failures after 10 years resulted from late endothelial failure. CONCLUSIONS: From 10 to 15 years after penetrating keratoplasty, the annual rate of endothelial cell loss was similar to that of normal corneas, corneal thickness increased, and late endothelial failure was the major cause of graft failure. PMID- 15747746 TI - Light-adjustable lens: development of in vitro nomograms. AB - PURPOSE: To determine whether digital spatial intensity patterns can be developed to effect precise in vitro correction of myopic, hyperopic, and astigmatic refractive errors in a silicone light-adjustable lens (LAL). Also, to determine whether a new spatial intensity pattern for "lock-in" is effective in vitro. METHODS: A digital interferometer/irradiation system was developed to irradiate LALs and measure the power change following irradiation. Light-adjustable lenses were mounted into a wet cell maintained at 35.0 +/- 0.5 degrees C (simulated ocular temperature) and allowed to equilibrate for a minimum of 2 hours. Ultraviolet light was then applied with spatial light intensity patterns to correct hyperopia, myopia, and astigmatism. Light-adjustable lenses were also treated to effect lock-in with a separate spatial light intensity pattern. Treated lenses were characterized for power change and optical quality. In the case of lock-in, exhaustive chemical extraction was also performed to determine the percentage of remaining macromer. RESULTS: Appropriate digital irradiation spatial intensity patterns were created to develop nomograms for in vitro correction of myopia, hyperopia, and astigmatism in approximate 0.25 D steps. Power changes were reproducible and did not alter optical quality of the LALs. Further, lock-in dosing of the LALs did not alter optical quality or significantly change LAL power. CONCLUSIONS: In vitro nomograms have been developed for a silicone LAL that permit precise correction of myopia, hyperopia, and astigmatism. Furthermore, a spatial light intensity pattern has been devised that effects lock-in without significantly altering LAL power or optical quality. PMID- 15747747 TI - Von Hippel-Lindau gene deletion and expression of hypoxia-inducible factor and ubiquitin in optic nerve hemangioma. AB - PURPOSE: Von Hippel-Lindau (VHL) disease is a hereditary cancer syndrome expressed in multiple organs caused by germline alterations of the VHL gene. We have shown VHL deletion in the "stromal" cells of retinal angiomas. The VHL protein-associated complex is a primary ubiquitin ligase for the ubiquitination of hypoxia-inducible factor (HIF). This study examines VHL and ubiquitin expression in optic nerve hemangiomas and juxtapapillary angiomas. METHODS: Using microdissection and polymerase chain reaction, four optic nerve hemangiomas (one also had juxtapapillary angioma) associated with VHL disease were analyzed for loss of heterozygosity in the VHL gene. In addition, expression of HIF and ubiquitin was evaluated in these tumors by immunohistochemistry. RESULTS: All informative optic nerve and juxtapapillary lesions showed loss of heterozygosity in the VHL gene detected in vacuolated "stromal" cells. Both HIF and ubiquitin were highly expressed in the hemangiomas of all four VHL cases. CONCLUSIONS: Like retinal angiomas and other VHL tumor lesions, VHL gene deletion is found in optic nerve hemangiomas and juxtapapillary angiomas. These tumor cells also express HIF and ubiquitin, the protein responsible for the negative regulation of HIF that results in the hypervascularization characteristic of VHL disease. PMID- 15747748 TI - A model of spectral filtering to reduce photochemical damage in age-related macular degeneration. AB - BACKGROUND/PURPOSE: Cumulative sunlight exposure and cataract surgery are reported risk factors for advanced age-related macular degeneration (AMD). Laboratory studies suggest that accumulation and photochemical reactions of A2E (N-retinylidene-N-retinylethanolamine) and its epoxides, components of lipofuscin, are important in AMD. To relate this data to the clinical setting, we modeled the effects of macular irradiance and spectral filtering on production of A2E and reactive oxygen intermediates (ROIs) in pseudophakic eyes with a clear or "yellow" intraocular lens (IOL) and in phakic eyes. METHODS: We calculated relative changes of macular irradiance as a function of light (390 to 700 nm) intensity, pupil size, age, and lens status, and modeled resulting all-trans retinal concentration and rates of production of A2E-related photochemicals and photon-induced ROIs in rods and retinal pigment epithelium (RPE). We compared these photoproducts following cataract surgery and IOL implantation with and without spectral sunglasses to normal age-related nuclear sclerotic lens changes. RESULTS: Following cataract and IOL surgery, all-trans-retinal and lipofuscin photochemistry would theoretically increase average generation of 1) A2E-related photochemicals, 2) ROI in rods and 3) ROI in RPE, respectively, 2.6-, 15- and 6.6 fold with a clear IOL, and 2.1-, 4.1- and 2.6 fold with a yellow IOL, but decrease approximately 30-, approximately 20- and 4-fold with a vermillion filter sunglass and clear IOL compared to an average 70 year old phakic eye. CONCLUSION: Sunglasses that strongly decrease both deep blue light and rod photobleaching, while preserving photopic sensitivity and color perception, would provide upstream protection from potential photochemical damage in subjects at risk for AMD progression after cataract surgery. PMID- 15747749 TI - Vitrectomy for epiretinal membranes with good visual acuity. AB - PURPOSE: To evaluate the visual results of vitrectomy for epiretinal membrane in eyes with a preoperative visual acuity of 20/50 or better. METHODS: The visual results and complications were analyzed following vitrectomy for idiopathic epiretinal membranes and epiretinal membranes secondary to retinal tears. This study was a retrospective, consecutive case series of 40 eyes of 40 patients treated by a single surgeon. RESULTS: The mean preoperative visual acuity was 20/50 +2 (range, 20/30 +1 to 20/50 -3). The mean visual acuity improved to 20/40 +2 (P = .02) by the final examination at a mean of 2.4 years following surgery. The status of the lens at the final examination was correlated with the visual results of surgery. Twenty-one eyes were phakic preoperatively, and 14 of these eyes had cataracts removed by the final examination. The mean preoperative visual acuity in seven eyes that were still phakic at the final examination was 20/50, and this decreased to 20/50 -2 (P = .82). The mean preoperative visual acuity was 20/50 +2 in 33 eyes that were pseudophakic by the final examination, and this improved to 20/32 -2 (P = .005). The visual acuity improved by 2 or more Snellen lines in only one in seven eyes (14%) that were still phakic on the final examination and in 14 of 33 eyes (42.4%) that were pseudophakic by the final examination. There were no serious surgical complications. CONCLUSIONS: Vitrectomy for epiretinal membranes is beneficial in eyes with relatively good preoperative visual acuities, but cataract surgery is necessary in phakic eyes to achieve long-term visual acuity improvement. PMID- 15747751 TI - Retinal pigment epithelium resurfacing of aged submacular human Bruch's membrane. AB - PURPOSE: To determine whether cultured fetal human retinal pigment epithelium (RPE) cells can attach and differentiate on submacular Bruch's membrane from donors over age 55. METHODS: Differential debridements of Bruch's membrane were performed to expose three different surfaces: the RPE basement membrane, the superficial inner collagenous layer (ICL) directly below the RPE basement membrane, and the deeper ICL. Approximately 3,146 cells/mm2 were seeded onto these Bruch's membrane explants and cultured for 1 or 7 days. Explants were bisected and examined histologically or analyzed with scanning electron microscopy. Nuclear density counts were performed on stained sections. Morphology and cell density were compared to those of cells seeded onto bovine corneal endothelial cell-extracellular matrix (BCE-ECM). RESULTS: Compared to cells seeded onto BCE-ECM at similar density, cell coverage and cellular morphology were poor at both time points. Unlike cells on BCE-ECM, cell density remained the same or decreased with time. In general, cell morphology on all surfaces worsened by day 7 compared to day 1. Although cells were more pigmented on RPE basement membrane and deep ICL at day 7, poor cellular morphology indicated the remaining cells were not well differentiated. An explant from a donor with large soft drusen showed the poorest resurfacing at day 7 in organ culture. CONCLUSIONS: These data indicate that aged submacular human Bruch's membrane does not support transplanted RPE survival and differentiation. The formation of localized RPE defects, cell death, and worsening cellular morphology on aged Bruch's membrane suggest that modification of Bruch's membrane may be necessary in patients with age-related macular degeneration receiving RPE transplants to prevent graft failure. PMID- 15747752 TI - Aggressive retinal astrocytomas in four patients with tuberous sclerosis complex. AB - OBJECTIVE: To report the clinical and histopathologic findings of retinal astrocytic tumors that showed progressive growth in four patients with tuberous sclerosis complex (TSC). METHODS: Four young children each developed an enlarging retinal neoplasm that eventually necessitated enucleation of the affected eye. The systemic findings, clinical course, and histopathologic findings were reviewed. RESULTS: Each patient had a progressively enlarging retinal mass associated with a total exudative retinal detachment and neovascular glaucoma. Enucleation was necessary in each case because the affected eye became blind and painful. The mean patient age at enucleation was 7 years, and the median age was 3 years. At the time of enucleation the tumors ranged from 10 to 20 mm in basal diameter and from 10 to 25 mm in thickness. Histopathologic studies of each eye revealed a giant cell astrocytoma that had produced a total exudative retinal detachment. The tumor cells showed positive immunoreactivity to neuron-specific enolase and glial fibrillary acidic protein. The retinal neoplasms in these cases were identical histopathologically to the subependymal giant cell astrocytoma that typifies TSC in the brain. One tumor filled the entire eye and perforated the globe. Although the lesions simulated retinoblastoma clinically, each patient had ocular and systemic findings of TSC, supporting the diagnosis of astrocytic hamartoma. CONCLUSIONS: Although retinal astrocytic lesions of TSC generally are stationary, they can sometimes grow relentlessly and cause severe ocular complications. Patients with retinal astrocytic hamartomas should have serial ophthalmic evaluations because of this possibility. PMID- 15747750 TI - Autism with ophthalmologic malformations: the plot thickens. AB - PURPOSE: To review the association of autism spectrum disorder (ASD) in individuals manifesting thalidomide embryopathy and Mobius sequence and compare them with three new studies in which ASD was also associated with ocular and systemic malformations: (1) a Swedish study of individuals with CHARGE association (Coloboma, Heart, choanal Atresia, developmental or growth Retardation, Genital anomaly, and Ear involvement); (2) a Swedish study of Goldenhar syndrome; and (3) Brazilian Mobius syndrome (sequence) study. METHODS: In the Swedish CHARGE study, 31 patients met the inclusion criteria (3+ or 4 of the common characteristics of the CHARGE syndrome). The same team of investigators also evaluated 20 Swedish patients with Goldenhar syndrome. In the Brazilian Mobius study, 28 children with a diagnosis of Mobius sequence were studied; some children had a history of exposure during their mother's pregnancy to the abortifacient drug misoprostol in an unsuccessful abortion attempt. RESULTS: In the CHARGE study, five patients had the more severe autism disorder and five had autistic-like condition. In the Goldenhar study, two had autism disorder and one had autistic-like condition. In the Brazilian Mobius study, the systemic findings of the misoprostol-exposed and misoprostol-unexposed patients were almost undistinguishable, and ASD was present in both groups (autism disorder in five and autistic-like condition in three). CONCLUSION: Autism spectrum disorder has been reported in two conditions with known early pregnancy exposure to the teratogenic agents thalidomide and misoprostol. In the Brazilian Mobius study, autism also occurred in both the misoprostol-exposed and misoprostol-unexposed groups. Autism also was present in patients with both CHARGE association and Goldenhar syndrome. PMID- 15747754 TI - Presumed sinus-related strabismus. AB - PURPOSE: To determine whether sinus disease may cause acquired strabismus. METHODS: Patients with idiopathic acquired (nonaccommodative) esotropia and/or hypotropia were questioned in detail about possible contributing factors (trauma; family history of strabismus; thyroid, neurologic, or rheumatologic disorders). Acute versus chronic onset was ascertained. Those without obvious cause of strabismus were investigated for possible sinus disease with sinus computed tomographic scan and otolaryngologic consultation. RESULTS: Over a period of 5 years, 59 patients were identified with sinus disease that correlated to their strabismus pattern(s). Twenty-three had "possible" sinus-related strabismus. They had sinus findings that correlated with the strabismus pattern (eg, hypotropia and adjacent maxillary sinus disease). Twenty-six had "likely" sinus-related strabismus. These patients had additional features, such as their own recognition that strabismus worsened along with sinus symptoms, or unusually severe sinus disease. Ten were diagnosed with "very likely" sinus-related strabismus. They had strong correlation between treatment of sinus disease and strabismus improvement. Eighteen patients required sinus surgery owing to failure of medical control. Age at onset of strabismus ranged from 6 months to 81 years. Forty patients required strabismus surgery. All had restriction of motility on forced duction testing under anesthesia. Control of sinus disease combined with range-of-motion eye exercise improved symptoms in 19 who did not require strabismus surgery. CONCLUSIONS: Occult sinus disease may cause acquired strabismus. Perhaps sinusitis leads to inflammation and secondary contracture in adjacent extraocular muscles. Although difficult to prove owing to the high frequencies of both strabismus and sinus disease, the association between the two may prove significant to strabismus treatment and long-term control. PMID- 15747753 TI - Human intraocular penetration pharmacokinetics of moxifloxacin 0.5% via topical and collagen shield routes of administration. AB - PURPOSE: To determine penetration of moxifloxacin 0.5% into human aqueous and vitreous via topical and collagen shield routes of administration. METHODS: Moxifloxacin 0.5% was administered prior to vitrectomy surgery through one of three routes: topical drops every 2 hours for 3 days, versus topical drops every 6 hours for 3 days, versus delivery using a 24-hour dissolvable cross-linked corneal collagen shield. Aqueous and vitreous moxifloxacin concentrations were assayed using high-performance liquid chromatography. RESULTS: Mean moxifloxacin concentrations in the every-2-hour group for aqueous (n = 9) and vitreous (n = 10) were 2.28 +/- 1.23 microg/mL and 0.11 +/- 0.05 microg/mL, respectively. Mean moxifloxacin concentrations in the every-6-hour group for aqueous (n = 10) and vitreous (n = 9) were 0.88 +/- 0.88 microg/mL and 0.06 +/- 0.06 microg/mL, respectively. Levels of minimum inhibitory concentration at which 90% of isolates are inhibited (MIC90) were far exceeded in the aqueous for a wide spectrum of pathogens that most commonly cause postoperative endophthalmitis. Moxifloxacin concentration in the vitreous did not exceed the MIC90 for several key organisms. Delivery of moxifloxacin via a collagen shield revealed a mean aqueous concentration of 0.30 +/- 0.17 microg/mL 4 hours after placement (n = 5). Vitreous levels at 4 hours, as well as aqueous and vitreous levels at 24 hours, were negligible using this route of administration. CONCLUSIONS: The findings of this investigation reveal that topically administered moxifloxacin 0.5% can achieve relatively high aqueous concentrations. Although aqueous moxifloxacin levels achieved through the use of a collagen shield delivery device are lower, there are several advantages to this route of delivery that make it appealing in the immediate postoperative period. Future studies will be needed to precisely define the role of fourth-generation fluoroquinolones and presoaked collagen shields in the prophylaxis or management of intraocular infections. PMID- 15747755 TI - Graded partial tenotomy of vertical rectus muscles for treatment of hypertropia. AB - PURPOSE: To evaluate the effectiveness of graded (adjustable intraoperatively) partial vertical rectus muscle tenotomy at the insertion in correcting small degrees of hypertropia. METHODS: All patients with best-corrected visual acuity of better than 6/30 in both eyes who over a 30-month period underwent partial tenotomy of vertical rectus muscle(s) only (no concurrent oblique muscles) were included. Improvement was evaluated 6 weeks postoperatively as change in alignment in prism diopters (PD) in primary gaze and in the field of action of the affected rectus muscle(s). Binocular function was evaluated by Titmus stereoacuity and the Worth 4-light tests. RESULTS: All 24 patients who met criteria for inclusion had diplopia preoperatively versus seven patients (29%) postoperatively (P < .005, Student's paired t test). Prisms were used by six preoperatively versus two postoperatively (P < .05, Student's paired t test). The average vertical deviation in primary gaze decreased from 8 PD to 2 PD (P < .005, Student's paired t test). In the field of action of the treated rectus muscle, hypertropia decreased from an average of 8 PD to 3 PD (P < .005, Student's paired t test). For the preoperative and the postoperative assessments available, stereoacuity improved after 10 (56%) of the 18 procedures and Worth 4-light testing showed improvement or maintenance of fusion after 15 (79%) of 19 procedures. CONCLUSIONS: Graded vertical rectus partial tenotomy can effectively reduce small degrees of hypertropia and associated diplopia, improve binocular function, and reduce or eliminate the need for prism correction. PMID- 15747756 TI - Value-based medicine and ophthalmology: an appraisal of cost-utility analyses. AB - PURPOSE: To ascertain the extent to which ophthalmologic interventions have been evaluated in value-based medicine format. METHODS: Retrospective literature review. Papers in the healthcare literature utilizing cost-utility analysis were reviewed by researchers at the Center for Value-Based Medicine, Flourtown, Pennsylvania. A literature review of papers addressing the cost-utility analysis of ophthalmologic procedures in the United States over a 12-year period from 1992 to 2003 was undertaken using the National Library of Medicine and EMBASE databases. The cost-utility of ophthalmologic interventions in inflation-adjusted (real) year 2003 US dollars expended per quality-adjusted life-year (dollars/QALY) was ascertained in all instances. RESULTS: A total of 19 papers were found, including a total of 25 interventions. The median cost-utility of ophthalmologic interventions was 5,219 dollars/QALY, with a range from 746 dollars/QALY to 6.5 million dollars/QALY. CONCLUSIONS: The majority of ophthalmologic interventions are especially cost-effective by conventional standards. This is because of the substantial value that ophthalmologic interventions confer to patients with eye diseases for the resources expended. PMID- 15747757 TI - Comparison of intraocular lens power calculation methods in eyes that have undergone laser-assisted in-situ keratomileusis. AB - PURPOSE: To compare methods of calculating intraocular lens (IOL) power for cataract surgery in eyes that have undergone myopic laser-assisted in-situ keratomileusis (LASIK). METHODS: Eleven eyes of eight patients who had previously undergone myopic LASIK (amount of LASIK correction, -5.50 +/- 2.61 D (SD); range, -8.78 to -2.38 D) and subsequently phacoemulsification with implantation of the SA60AT IOLs were included (refractive error after cataract surgery, -0.61 +/- 0.79 D; range, -2.0 to 1.0 D). We evaluated the accuracy of various combinations of (1) single-K versus double-K (in which pre-LASIK keratometry is used to estimate effective lens position) versions of the IOL formulas; the Feiz-Mannis method was also evaluated; (2) four methods for calculating corneal refractive power (clinical history, contact lens overrefraction, adjusted EffRP (EffRP(adj)), and Maloney methods); and (3) four IOL formulas (SRK/T, Hoffer Q, Holladay 1, and Holladay 2). The IOL prediction error was obtained by subtracting the IOL power calculated using various methods from the power of the implanted IOL, and the F test for variances was performed to assess the consistency of the prediction performance by different methods. RESULTS: Compared to double-K formulas, single-K formulas predicted lower IOL powers than the power implanted and would have left patients hyperopic in the majority of the cases; the Feiz Mannis method had the largest variance. For the Hoffer Q and Holladay 1 formulas, the variances for EffRP(adj) were significantly smaller than those for the clinical history method (0.43 D2 vs 1.74 D2, P = .018 for Hoffer Q; 0.75 D2 vs 2.35 D2, P = .043 for Holladay 1). The Maloney method consistently underestimated the IOL power but had significantly smaller variances (0.19 to 0.55 D2) than those for the clinical history method (1.09 to 2.35 D2) (P < .015). There were no significant differences among the variances for the four formulas when using each corneal power calculation method. CONCLUSIONS: The most accurate method was the combination of a double-K formula and corneal values derived from EffRP(adj). The variances in IOL prediction error were smaller with the Maloney and EffRP(adj) methods, and we propose a modified Maloney method and second method using Humphrey data for further evaluation. PMID- 15747758 TI - Efficacy and efficiency of a new involutional ptosis correction procedure compared to a traditional aponeurotic approach. AB - PURPOSE: This was a retrospective study to compare the efficacy and efficiency of a new small anterior incision, minimal dissection ptosis procedure with that of a traditional anterior aponeurotic approach for the correction of aponeurotic ptosis. METHODS: The results of a chart and photograph review of 36 patients with 49 ptotic eyelids who had ptosis correction by a small-incision, minimal dissection procedure were compared with those of 36 patients with 49 ptotic eyelids who had ptosis correction by a traditional aponeurotic approach. RESULTS: The successful correction of the eyelid height and the rate of recommendation for reoperation were not significantly different for the 49 lids corrected in each arm of the study. The incidence of attaining good eyelid contour was significantly better in the small-incision group, where 41 (97.6%) of 42 lids evaluated by photographs had good contour compared with 29 (78.4%) of 37 lids in the traditional group. Operating time per lid was significantly less for the small-incision, minimal dissection group, 25.3 +/- 13.0 minutes (range, 13 to 68 minutes), compared with 55.4 +/- 16.6 minutes (range, 35 to 119) for the traditional group. CONCLUSIONS: Compared with the traditional aponeurotic approach, the new small-incision, minimal dissection technique for ptosis correction is equally efficacious in correcting eyelid height, superior in producing desirable eyelid contour, and much quicker to perform. PMID- 15747760 TI - Does medical treatment influence the success of trabeculectomy? AB - PURPOSE: Many ophthalmologists believe that long-term use of topically applied glaucoma medications can adversely affect results of fistulizing surgery. This presentation critically analyzes the published studies most often cited in support of this view to determine whether this conclusion is justified. METHODS: Morphologic effects of long-term treatment with antiglaucoma drugs on the conjunctiva and Tenon's capsule in glaucomatous patients have been studied. The results of these studies encouraged investigators to examine the influence of prior therapy on the success of trabeculectomy performed in patients with open angle glaucoma. From this work, many have concluded that long-term use of topically applied glaucoma medications can adversely affect the results of fistulizing surgery. These results and conclusions are summarized and critically analyzed to determine whether this conclusion is justified. RESULTS: Morphologic studies describe increased numbers of macrophages, fibroblasts, lymphocytes, and mast cells in conjunctival and Tenon's capsule specimens taken from patients receiving long-term antiglaucoma drugs. These findings suggest a potential for more inflammation and subsequent scarring following trabeculectomies in these patients. Efforts to confirm the clinical relevance of these histologic findings in open-angle glaucoma patients with a history of long-term antiglaucoma medication prior to surgery have been published. These retrospective, nonrandomized, unmasked studies of open-angle glaucoma patients include treatment groups and surgeries that are not comparable. In addition, the medical treatments within these studies do not reflect our current approaches to the medical management of open-angle glaucoma. CONCLUSIONS: At present, there is no convincing clinical evidence that long-term medical treatments influence the success of contemporary trabeculectomy surgery performed on open-angle glaucoma patients. PMID- 15747759 TI - Vascular perfusion of choroidal melanoma by 3.0 tesla magnetic resonance imaging. AB - PURPOSE: Because signal-to-noise performance improves with increased magnetic field strength, the quality of magnetic resonance images is greater at 3.0 tesla (T) than at 1.5 T. Because of the longer T1 values at higher field strength, intravenously administered magnetic resonance contrast agents provide improved T1 enhancement at 3.0 T. We have used these factors to obtain high-quality contrast enhanced imaging of small intraocular lesions using a standard head radiofrequency volume coil. Specifically, we have examined lesion size and magnitude of maximum contrast enhancement in a series of intraocular melanomas before and during therapy. METHODS: Eighteen patients with intraocular masses were examined by 3.0 T magnetic resonance imaging (MRI) including intravenous contrast enhancement. Precontrast images were acquired through the orbits followed by sequential postcontrast images at 1-minute intervals for 5 minutes. The magnitude of contrast enhancement of the lesion, extraocular muscles, and brain parenchyma was measured as a percentage increase in magnetic resonance signal over the preenhancement signal intensity. RESULTS: Lesions demonstrated different levels of enhancement ranging up to 130%. Three patterns of enhancement -0% to 20%, 20% to 50%, and >50%-were identified. Brain parenchyma, benign lesions, and responsive tumors following brachytherapy with 125I demonstrated enhancement of less than 20%. Four choroidal melanomas showed intermediate (20% to 50%) levels of enhancement. Four malignant lesions (three melanomas, one metastatic tumor), as well as the extraocular muscles, showed strong, rapid enhancement (>50%). Four patients who had MRI studies before and following plaque brachytherapy ultimately demonstrated a decline in the contrast enhancement following treatment. CONCLUSIONS: Contrast enhancement of intraocular lesions measured by 3.0 T MRI demonstrates different patterns of enhancement that may be useful for indicating the degree of malignancy and in monitoring response to therapy. PMID- 15747761 TI - The parameters of informed consent. AB - PURPOSE: To describe the components of a proper informed consent; which risks must be disclosed and which need not; additional safeguards for incapacitated persons, minors, and research subjects; and where the law will imply consent that is not otherwise obtained. METHODS: Summarization of current law obtained from legal treatises, reports of recent cases, and personal experience as a reviewer and expert. RESULTS: Lack of informed consent can reinforce a claim of medical malpractice or serve as an alternative point of attack when the case is otherwise weak. Special requirements must be met when patients are the subjects of clinical research. CONCLUSION: Demonstration of a well-conducted process, not merely of a paper, not only protects the physician from exposure to liability, but increases the patient's autonomy in decisions concerning health and encourages compliance with treatment. PMID- 15747762 TI - Final results of the Early Treatment for Retinopathy of Prematurity (ETROP) randomized trial. AB - PURPOSE: To present the final results of the Early Treatment for Retinopathy of Prematurity Study. METHODS: Infants with bilateral high-risk prethreshold retinopathy of prematurity (ROP) (n = 317) had one eye randomized to early retinal ablative treatment and the fellow eye managed conventionally (control eye). In asymmetric cases (n = 84), the eye with high-risk prethreshold ROP was randomized to early or to conventional management. High risk was determined using a model based on the Cryotherapy for Retinopathy of Prematurity natural history cohort. The primary outcome was visual acuity assessed by masked testers using the Teller acuity card procedure. Structural examinations were performed at 6 and 9 months. RESULTS: Grating acuity results showed a reduction in unfavorable visual acuity outcomes with earlier treatment, from 19.8% to 14.3% (P < .005). Unfavorable structural outcomes were reduced from 15.6% to 9.0% (P < .001) at 9 months. Further analysis supported retinal ablative therapy for eyes with type I ROP, defined as zone I, any stage ROP with plus disease; zone I, stage 3 ROP without plus disease; or zone II, stage 2 or 3 with plus disease. The analysis supported a "wait and watch" approach to type II ROP, defined as zone I, stage 1 and 2 without plus disease, or zone II, stage 3 without plus disease. These eyes should be considered for treatment only if they progress to type I ROP or threshold. CONCLUSION: Early treatment of high-risk prethreshold ROP significantly reduced unfavorable outcomes in both primary and secondary (structural) measures. PMID- 15747763 TI - Effects of latrunculin B on outflow facility, intraocular pressure, corneal thickness, and miotic and accommodative responses to pilocarpine in monkeys. AB - PURPOSE: To determine if low doses of topical latrunculin B (LAT-B) will increase outflow facility and decrease intraocular pressure (IOP) without adversely affecting the cornea, and inhibit miotic and accommodative responses to pilocarpine, in ocular normotensive monkeys. METHODS: Intraocular pressure was measured by Goldmann tonometry before and after one and nine dose(s) of 0.005% and 0.01% topical LAT-B/vehicle given twice daily on successive weeks. Outflow facility was then measured by perfusion following 15 doses. Central corneal thickness was measured by ultrasonic pachymetry before and after one and nine dose(s) of 0.01% LAT-B/vehicle. Pupillary diameter (calipers) and accommodation (refractometry) before and after one dose of 0.005% and 0.02% LAT-B were determined. RESULTS: LAT-B dose-dependently decreased IOP, multiple doses more than a single dose. Maximal hypotension after one dose was 2.5 +/- 0.3 mm Hg (0.005% LAT-B; n = 8; P < .001) or 2.7 +/- 0.6 mm Hg (0.01% LAT-B; n = 8; P < .005); maximal hypotension after nine doses was 3.2 +/- 0.5 mm Hg (0.005% LAT-B; n = 8; P < .001) or 4.4 +/- 0.6 mm Hg (0.01% LAT-B; n = 8; P < .001). Outflow facility was increased by 75 +/- 13% (n = 7; P < .005). Central corneal thickness was not changed after one or nine dose(s) of 0.01% LAT-B. The miotic and accommodative responses to intramuscular pilocarpine were dose-dependently inhibited. At 0.02% LAT-B, the inhibition of miosis was essentially complete when compared with the pre-LAT-B value, whereas the inhibition of accommodation was only about 25%. At 0.005% LAT-B, the effects were trivial. CONCLUSIONS: In ocular normotensive monkeys, 0.005/0.01% LAT-B administered topically increases outflow facility and/or decreases IOP, but does not affect the cornea. Multiple doses reduce IOP more than a single dose. LAT-B dose-dependently relaxes the iris sphincter and ciliary muscle, with some separation of the miotic and accommodative effects. PMID- 15747764 TI - Computerized expert system for evaluation of automated visual fields from the Ischemic Optic Neuropathy Decompression Trial: methods, baseline fields, and six month longitudinal follow-up. AB - PURPOSE: To validate a computerized expert system evaluating visual fields in a prospective clinical trial, the Ischemic Optic Neuropathy Decompression Trial (IONDT). To identify the pattern and within-pattern severity of field defects for study eyes at baseline and 6-month follow-up. DESIGN: Humphrey visual field (HVF) change was used as the outcome measure for a prospective, randomized, multicenter trial to test the null hypothesis that optic nerve sheath decompression was ineffective in treating nonarteritic anterior ischemic optic neuropathy and to ascertain the natural history of the disease. METHODS: An expert panel established criteria for the type and severity of visual field defects. Using these criteria, a rule-based computerized expert system interpreted HVF from baseline and 6-month visits for patients randomized to surgery or careful follow up and for patients who were not randomized. RESULTS: A computerized expert system was devised and validated. The system was then used to analyze HVFs. The pattern of defects found at baseline for patients randomized to surgery did not differ from that of patients randomized to careful follow-up. The most common pattern of defect was a superior and inferior arcuate with central scotoma for randomized eyes (19.2%) and a superior and inferior arcuate for nonrandomized eyes (30.6%). Field patterns at 6 months and baseline were not different. For randomized study eyes, the superior altitudinal defects improved (P = .03), as did the inferior altitudinal defects (P = .01). For nonrandomized study eyes, only the inferior altitudinal defects improved (P = .02). No treatment effect was noted. CONCLUSIONS: A novel rule-based expert system successfully interpreted visual field defects at baseline of eyes enrolled in the IONDT. PMID- 15747765 TI - Intravitreal human immune globulin in a rabbit model of Staphylococcus aureus toxin-mediated endophthalmitis: a potential adjunct in the treatment of endophthalmitis. AB - OBJECTIVES: To test the feasibility of human immune globulin (IG, Gamimune N, 10%) as a new treatment for endophthalmitis, the ocular tolerance, distribution, and ability of intravitreal IG to attenuate the toxic effects of Staphylococcus aureus culture supernatant were evaluated in a rabbit model. METHODS: Effects of intravitreally injected IG were assessed histologically and with Western blot analysis performed 1 to 5 days after injection. IG reactivity to products of S. aureus strain RN4220 was tested by Western blotting, using known toxins (beta hemolysin and toxic shock syndrome toxin-1) and a concentrated culture supernatant containing S. aureus exotoxins (pooled toxin, PT). Endophthalmitis was induced by intravitreal PT injection. For treatment, IG and PT were mixed and injected simultaneously, or IG was injected immediately after, or 6 hours after, PT injection. PT toxicity was graded clinically and histologically over 9 days. RESULTS: IG persisted intravitreally at least 5 days, inducing no clinical inflammation and minimal mononuclear cell infiltration. In the endophthalmitis model, toxicity from PT was significantly reduced when IG was mixed with PT and injected simultaneously, or when IG was delivered immediately after PT. Only minimal clinically detectable reductions were observed when IG delivery was delayed 6 hours. CONCLUSIONS: Intravitreal IG is well tolerated in the rabbit eye and attenuates the toxicity of culture supernatant containing S. aureus exotoxins. Because toxin elaboration likely occurs gradually in true infection, reduced effects observed with delayed treatment in this toxin-injected model do not preclude clinical application. IG may represent a novel adjunct in endophthalmitis treatment. PMID- 15747766 TI - The accuracy of digital-video retinal imaging to screen for diabetic retinopathy: an analysis of two digital-video retinal imaging systems using standard stereoscopic seven-field photography and dilated clinical examination as reference standards. AB - PURPOSE: To evaluate the accuracy of two digital-video retinal imaging (DVRI) systems to detect diabetic retinopathy. METHODS: A prospective, masked, technology assessment was conducted for two DVRI systems at a tertiary care Veterans Affairs Medical Center. Group A (n = 151 patients) was imaged with a 640x480 resolution system and group B (n = 103 patients) with an 800x600 resolution system. Four retinal evaluations were performed on each patient: DVRI with undilated pupils using one imaging field (U-DVRI), DVRI with dilated pupils using three imaging fields (D-DVRI), dilated clinical examination, and Early Treatment Diabetic Retinopathy Study stereoscopic seven-field photography (ETDRS P). Two analyses of accuracy were conducted, one using ETDRS-P as a "gold standard" (ETDRS-GS) and one using dilated clinical examination as a "gold standard" (C-GS). RESULTS: For group A, using the ETDRS-GS, sensitivities of U DVRI and D-DVRI were 0.66 and 0.66; specificities of U-DVRI and D-DVRI were 0.66 and 0.86. Using the C-GS, sensitivities of U-DVRI and D-DVRI were 0.79 and 0.80; specificities of U-DVRI and D-DVRI were 0.68 and 0.85. For group B, using the ETDRS-GS, sensitivities of U-DVRI and D-DVRI were 0.76 and 0.85; specificities of U-DVRI and D-DVRI were 0.45 and 0.80. Using the C-GS, sensitivities of U-DVRI and D-DVRI were 0.81 and 0.87; specificities of U-DVRI and D-DVRI were 0.45 and 0.69. For both groups, dilation significantly improved specificities. CONCLUSIONS: The 800x600 resolution DVRI system offers an accurate method of detecting diabetic retinopathy, provided there is adequate pupillary dilation and three retinal images are taken. DVRI technology may help facilitate retinal screenings of growing diabetic populations. PMID- 15747768 TI - Congenital abnormalities of cranial nerve development: overview, molecular mechanisms, and further evidence of heterogeneity and complexity of syndromes with congenital limitation of eye movements. AB - PURPOSE: The clinical and molecular genetic classification of syndromes with congenital limitation of eye movements and evidence of cranial nerve dysgenesis continues to evolve. This monograph details clinical and molecular genetic data on a number of families and isolated patients with congenital fibrosis of the extraocular muscles (CFEOM) and related disorders, and presents an overview of the mechanisms of abnormal patterns of motor and sensory cranial nerve development in these rare syndromes. METHODS: Clinical examination of one patient with CFEOM1, one family with clinical features of CFEOM2, one family with recessive CFEOM3, one family with horizontal gaze palsy and progressive scoliosis (HGPPS), and four patients with various combinations of congenital cranial nerve abnormalities. Genotyping of families with CFEOM and HGPPS for polymorphic markers in the regions of the three known CFEOM loci and in the HGPPS region, and mutation analysis of the ARIX and KIF21A genes in patients with CFEOM were performed according to standard published protocols. RESULTS: The patient with CFEOM1 had the second most common mutation in KIF21A, a 2861 G>A mutation that resulted in an R954Q substitution. The family with CFEOM2 phenotype did not map to the CFEOM2 locus. The family with recessive CFEOM3 did not map to any of the known loci. The HGPPS family mapped to 11q23-q25. One patient had optic nerve hypoplasia and fifth nerve dysfunction. Two patients had the rare combination of Mobius syndrome and CFEOM. One patient had Mobius syndrome and fifth nerve dysfunction. CONCLUSIONS: There is genetic heterogeneity in CFEOM2 and CFEOM3. Abnormalities in sensory nerves can also accompany abnormalities of motor nerves, further substantiating the effect of individual mutations on developing motor as well as sensory cranial nerve nuclei. PMID- 15747769 TI - Anterior lens capsule management in pediatric cataract surgery. AB - PURPOSE: To describe and analyze pediatric anterior capsulotomy techniques and make recommendations. METHODS: Five anterior capsulotomy techniques were compared using a porcine model. Extensibility was measured by calculating the mean stretch to-rupture circumference of each capsulotomy (20 eyes per technique) as a percentage of its circumference at rest. Edge characteristics were reviewed using scanning electron microscopy. A 10-year review of consecutive pediatric cataract surgeries performed by the author focused on the anterior capsulotomy results. A worldwide survey was used to determine current practice patterns. RESULTS: Manual continuous curvilinear capsulorrhexis (CCC) produced the most extensible porcine capsulotomy (185%) with the most regular edge and is preferred by surgeons for patients aged 2 years and older. In the pseudophakic clinical cases reviewed, a radial tear developed in 3 (6.5%) of 46 manual CCC cases. Vitrectorhexis (porcine extensibility, 161%) is preferred by surgeons during the first 2 years of life. A radial tear developed in 16 (7.7%) of 208 vitrectorhexis pseudophakic eyes (29 tears in 284 pseudophakic eyes [10.2%] overall). The Kloti diathermy unit, Fugo plasma blade, and "can-opener" technique produced porcine capsulotomies of 145%, 170%, and 149% extensibility, respectively, and radial tears numbering 4 (21%) of 19, 5 of 8, and 1 of 2, respectively, in the clinical series. CONCLUSIONS: All five capsulotomy techniques are recommendable for children. Only the vitrectorhexis and manual CCC are commonly used today. Vitrectorhexis is well suited for use in infants and young children; manual CCC is best used beyond infancy, and it produces the most stable edge. PMID- 15747771 TI - Gender, politics, and radioactivity research in interwar Vienna: the case of the Institute for Radium Research. AB - This essay explores the significance of political and ideological context as well as experimental culture for the participation of women in radioactivity research. It argues that the politics of Red Vienna and the culture of radioactivity research specific to the Viennese setting encouraged exceptional gender politics within the Institute for Radium Research in the interwar years. The essay further attempts to provide an alternative approach to narratives that concentrate on personal dispositions and stereotypical images of women in science to explain the disproportionately large number of women in radioactivity research. Instead, the emphasis here is on the institutional context in which women involved themselves in radioactivity in interwar Vienna. This approach places greater importance on contingencies of time and place and highlights the significance of the cultural and political context in a historical study while at the same time shedding light on the interrelation between scientific practices and gender. PMID- 15747767 TI - Photorefractive keratectomy for anisometropic amblyopia in children. AB - PURPOSE: To assess the safety and efficacy of photorefractive keratectomy (PRK) in children with anisometropic amblyopia and to define the characteristics of children who may be candidates for PRK. METHODS: This thesis comprises four parts: (1) a retrospective analysis of risk factors predictive of amblyopia treatment failure in 104 children, (2) a prospective study of pachymetry in 198 eyes of 108 children, (3) development and implementation of a protocol to perform PRK under general anesthesia, and (4) a prospective interventional case comparison study of PRK in 11 noncompliant children with anisometropic amblyopia to evaluate safety and long-term outcomes. Compliant and noncompliant children with anisometropic amblyopia were analyzed as controls. RESULTS: Factors associated with conventional anisometropic amblyopia treatment failure were poor compliance (P = .004), age 6 years or older (P = .01), astigmatism > or = 1.5 diopters (P = .0002), and initial visual acuity of 20/200 or worse (P = .02). Central and paracentral pachymetry measurements were similar to published adult values. The general anesthesia protocol was efficient, and the laser functioned properly in all cases. All children did well with no anesthesia-related or treatment-related complications. Two years following PRK, the mean reduction in refractive error was 9.7 +/- 2.6 diopters for myopes (P = .0001) and 3.4 +/- 1.3 diopters for hyperopes (P = .001). The cycloplegic refractive error in 9 of 11 treated eyes was within 3 diopters of that in the fellow eye. Uncorrected visual acuity in the amblyopic eye improved by > or = 2 lines in seven of nine children; best-corrected visual acuity improved by > or = 2 lines in six of nine children. Stereopsis improved in five of nine children. The mean visual acuity of the PRK patients at last follow-up was significantly better than that of noncompliant controls (P = .003). The safety and efficacy indices for PRK in this study were 1.24 and 1.12, respectively. CONCLUSIONS: Photorefractive keratectomy can be safely performed in children with anisometropic amblyopia. Visual acuity and stereopsis improved in most eyes, even in older children. Photorefractive keratectomy may have an important role in the management of anisometropic amblyopia in noncompliant children. PMID- 15747772 TI - Racial science in social context: John R. Baker on eugenics, race, and the public role of the scientist. AB - In 1974 a British biologist, John Randal Baker (1900-1984), published a large and controversial book simply entitled Race that reiterated persistent eugenicist themes concerning the relation between race, intelligence, and progress. The history of Baker's book is a case study in the politics of scientific publishing, and his ideas influenced scholars associated with later works such as The Bell Curve. Baker, a student of Julian Huxley, was a longtime participant in the British eugenics movement and opponent of what he took to be a facile belief in human equality. In 1942, together with Michael Polanyi, he founded the Society for Freedom in Science to oppose those who advocated the central planning of scientific research. Baker's eugenics, political activities, and views on race express an elitist individualism, associated with the conservative wing of the eugenics movement, that this paper explores in the context of his career as a whole. PMID- 15747774 TI - The multistage model of cancer development: some implications. AB - The multistage model, introduced by Armitage and Doll, was very successful at describing many features of cancer development. Doll and Peto noted a significant departure below the prediction of the model and suggested that this could be due to undercounting of cases at older ages, or to the 'biology of extreme old age.' Moolgavkar pointed out that it could also be due to the approximation used. The recent observation that cancer incidence falls rapidly above age 80 has stimulated new modelling investigations, such as the Pompei-Wilson beta model (which does reproduce the rapid fall). In the present paper, we argue that Moolgavkar's criticisms, while mathematically correct, do not affect the conclusions, particularly the constancy of the number of stages across different cancer registries (Cook, Doll and Fellingham. 1969: A mathematical model for the age distribution of cancer in man. International Journal of Cancer 4, 93-112). We discuss several exact solutions, compare them with the most recent data, and prove rigorously that the standard Armitage-Doll multistage model can never reproduce the sharp turnaround in cancer incidence at old age seen in the data. We discuss in detail multistage processes which have a property observed in many laboratory studies, namely that some stages progress much faster than the others. We verify mathematically the intuition that sufficiently fast stages do not appreciably affect the incidence rate of cancer, and discuss implications of this fact for cancer treatment strategies. We also show that the simplest possible modification of the Armitage-Doll model to incorporate cellular senescence just leads to the Pompei-Wilson beta model. PMID- 15747775 TI - 5-HT loss in rat brain by type II pyrethroid insecticides. AB - STUDY OBJECTIVE: Type II pyrethroids are a group of insecticides largely used in agriculture and public health. The nervous system is the main target for pyrethroids in insects and mammals. One notable form of toxicity associated with over exposure has been a facial cutaneous paraesthesia and irritation-related respiration symptoms including behavioural excitation mainly observed in workers spraying pyrethroids or in occupational settings. In acutely exposed rats, type II pyrethroids produce a severe syndrome characterized by salivation and choreoathetosis. Because many of the acute functional effects of type II pyrethoids can be associated with the neurotoxic effect on 5-hydroxytryptamine (5 HT) neurones, the objective of the present study was to examine whether deltamethrin, cyfluthrin and lambda-cyhalothrin administration results in changes of 5-HT content in rat brain. Characterizing this target will help us to better understand the toxicological effects of type II pyrethroids. DESIGN: Rats were injected with either corn oil or pyrethroids (deltamethrin, 20 mg/kg per day, i.p., for 6 days; cyfluthrin, 14 mg/kg per day, i.p., for 6 days; lambda cyhalothrin, 8 mg/kg per day, i.p., for 6 days). The frontal cortex, hippocampus, midbrain and striatum were removed at 24 hours post treatment and were analysed for content of 5-HT and 5-HIAA using a HPLC method with electrochemical detection. RESULTS: A serotonin depleting effect was produced by these type II pyrethroids. The concentration of 5-HT and its metabolite 5-HIAA decreased in the brain regions from pyrethroid treated animals. Pyrethroids accelerated the turnover of 5-HT in midbrain and striatum areas. It is concluded that pyrethroids affect serotonin neurotransmission. PMID- 15747770 TI - Dissecting the genetics of human high myopia: a molecular biologic approach. AB - PURPOSE: Despite the plethora of experimental myopia animal studies that demonstrate biochemical factor changes in various eye tissues, and limited human studies utilizing pharmacologic agents to thwart axial elongation, we have little knowledge of the basic physiology that drives myopic development. Identifying the implicated genes for myopia susceptibility will provide a fundamental molecular understanding of how myopia occurs and may lead to directed physiologic (ie, pharmacologic, gene therapy) interventions. The purpose of this proposal is to describe the results of positional candidate gene screening of selected genes within the autosomal dominant high-grade myopia-2 locus (MYP2) on chromosome 18p11.31. METHODS: A physical map of a contracted MYP2 interval was compiled, and gene expression studies in ocular tissues using complementary DNA library screens, microarray matches, and reverse-transcription techniques aided in prioritizing gene selection for screening. The TGIF, EMLIN-2, MLCB, and CLUL1 genes were screened in DNA samples from unrelated controls and in high-myopia affected and unaffected family members from the original seven MYP2 pedigrees. All candidate genes were screened by direct base pair sequence analysis. RESULTS: Consistent segregation of a gene sequence alteration (polymorphism) with myopia was not demonstrated in any of the seven families. Novel single nucleotide polymorphisms were found. CONCLUSION: The positional candidate genes TGIF, EMLIN 2, MLCB, and CLUL1 are not associated with MYP2-linked high-grade myopia. Base change polymorphisms discovered with base sequence screening of these genes were submitted to an Internet database. Other genes that also map within the interval are currently undergoing mutation screening. PMID- 15747776 TI - Dose-dependent transcriptome changes by metal ores on a human acute lymphoblastic leukemia cell line. AB - The increased morbidity of childhood leukemia in Fallon, Nevada and Sierra Vista, Arizona has prompted great health concern. The main characteristic that these two towns share is the environmental pollution attributed to metal ore from abandoned mining operations. Consequently, we have investigated the transcriptome effects of metal ores from these endemic areas using a human T-cell acute lymphoblastic leukemia cell line (T-ALL). Metal ore from Fallon significantly increased cell growth after 24, 48 and 72 h of incubation at 1.5 microg/mL concentration, as measured by trypan-blue. Sierra Vista ore significantly increased cell growth with 0.15 and 1.5 microg/mL following 72 h of incubation. From human cDNA microarray, results indicate that in total, eight genes, mostly metallothionein (MT) genes, were up-regulated and 10 genes were down-regulated following treatment of the T-ALL cells with 0.15 and 1.5 microg/mL of metal ores at 72 h, in comparison with untreated cells. Twenty-eight metals of both ores were quantified and their presence may be associated with the cell growth rate and dose-dependent activation of transcriptomes in immature T-cells. PMID- 15747777 TI - Systemic effects and skin injury after experimental dermal exposure to monochloroacetic acid. AB - There have been many fatal occupational accidents of skin exposure to monochloroacetic acid (MCA). However, there have been no reports of dermatological findings and the lethal consequences have not yet been demonstrated. Therefore, harmful local and systemic effects were investigated after dermal exposure to MCA. A 0.5 mL aliquot of MCA solution (40% w/w) was applied to the abdominal skin of ten 10-week-old male SD rats under anesthesia. The exposure area (25 x 25 mm2) was 1.6% of the total surface area. The dose of MCA per area was 34.1 mg/cm2. Saline was similarly administered to 10 control rats. Histopathological findings after 10 min were observed by light microscopy. Blood samples were collected by exsanguinations from the carotid arteries after 4 h. Skin samples were collected 10 min after the initial exposure. Histological findings showed severe degeneration of collagen bundles in the epidermis and subcutaneous tissues. P(CO2), HCO(3)-, TCO2, BE and glucose levels were decreased in the MCA group. AST, m-AST, ALT, BUN, Cr, NH3, lactic acid, pyruvic acid, RBC, Hb, Hct, total protein and albumin were increased in the MCA group. The burn was determined to be a third-degree burn on the basis of the histopathological findings. The severe toxicity was probably a consequence of the rapid permeability. Biochemical parameters were a consequence of hepatocellular injuries, renal dysfunction, dysglyconeogenesis and dysfunction of ammonia metabolism. MCA reportedly enters the TCA cycle and inhibits aconitase. MCA metabolites also inhibit pyruvate carboxylase in the gluconeogenesis pathway. Therefore, the important serum biochemical abnormalities such as hypoglycemia and lactic acidosis should be monitored to find the acute systemic disorders. PMID- 15747779 TI - Cement: a common cancer agent? PMID- 15747778 TI - Corticosterone in drinking water: altered kinetics of a single oral dose of corticosterone and concentrations of plasma sodium, albumin, globulin, and total protein. AB - Effects of chronic exposure to corticosterone in drinking water on corticosterone kinetics, blood chemistry, and concentrations of catecholamines in parts of brain were studied in Long-Evans rats. Rats were randomly grouped into 3 x 2 treatments (n=4), with three treatments of drinking water (tap water, or 2.5% ethanol, or 400 microg/mL of corticosterone in 2.5% ethanol) for 28 days and two treatments of gavage with a single dose of either corn oil or corticosterone 20 mg/kg on day 28. Blood samples were collected at 0, 15, 30, 60, 120, 240, 480, and 720 min after dosing to determine plasma corticosterone concentrations. Blood samples were collected for clinical pathology on day 42. Hippocampus, cerebral cortex, caudate-putamen, and pons were examined to determine concentrations of catecholamines and activities of esterases. Concentrations of plasma corticosterone before gavage of the corticosterone-drinking rats (47.61 +/- 1.13 ng/mL) were lower than the water (418.47 +/- 1.13 ng/mL) or the ethanol rats (383.71 +/- 1.13 ng/mL, P < 0.0001). Plasma corticosterone rose to peak concentrations by 15 min after gavage in all three groups of drinking rats. Corticosterone-drinking rats had concentrations of plasma corticosterone that returned to basal levels slower than water- and ethanol-drinking rats. Plasma sodium and chloride concentrations were lower in the corticosterone-drinking rats than the water-drinking rats (P < 0.01). Plasma albumin, globulin, and total protein were highest in the corticosterone-drinking rats when compared to the other groups of drinking rats (P < 0.001, P < 0.05, and P < 0.001, respectively). Corticosterone in drinking water did not affect activities of brain neurotoxic esterase, carboxylesterase, acetylcholinesterase, or concentrations of monoamines and their metabolites. A single oral dose of corticosterone reduced neurotoxic esterase activity in the cerebral cortex (P < 0.05) and increased norepinephrine concentrations in the hippocampus (P < 0.05). PMID- 15747780 TI - New Medicare screening test coverage. PMID- 15747781 TI - Importance of transplantation history in ABO discrepancies. PMID- 15747782 TI - Relationships among patient age, diagnosis, hospital type, and clinical laboratory utilization. AB - OBJECTIVE: The aging population will likely have a major impact on laboratory utilization. Utilization data will be necessary for laboratory managers to make informed decisions concerning staffing patterns and services offered. DESIGN: In a retrospective non-descriptive study, the relationships among age groups, hospital type, diagnosis, and the numbers and types of laboratory tests performed were investigated. SETTING: Half of the participants were from a private hospital, Touro Infirmary, and half were from a large public hospital, The Medical Center of Louisiana at New Orleans. Both facilities are located in New Orleans, Louisiana. PATIENTS: Laboratory records from a random sample of 250 inpatients age 21 to 64, a sample from 250 inpatients age 65 to 84, and a sample from 250 inpatients age 85 and over with at least one of five admission or discharge diagnoses were analyzed. INTERVENTIONS: Twenty-five records from each of the five diagnostic categories for each of the three age groups and two hospital types were analyzed, yielding a total sample of 750 records. MAIN OUTCOME MEASURES: Laboratory tests for each inpatient stay were counted and categorized for analysis. The one-way ANOVA was used to test the degree of concordance between age groups and numbers of tests ordered and between age groups and types of tests ordered across hospital types. RESULTS: Data analysis showed statistically significant differences in the total number of laboratory tests ordered for the three age groups regardless of facility (p 0.008). The age group with the highest number of total laboratory tests ordered was the group aged 65 to 84 (48.64 mean tests per patient). Across the total sample, more tests were ordered at the public facility than the private facility (51.75 and 32.42 mean tests per patient, respectively). Statistically significant differences in orders between the two facilities were noted in chemistry, hematology, and toxicology (p < 0.001). When analyzing numbers of tests by age group and facility, no statistically significant differences were noted in any laboratory category. Analysis of disease and laboratory test categories, regardless of facility, showed statistically significant differences in numbers of tests ordered in microbiology, cytology, histology (p < 0.001), and blood bank (p 0.001). When analyzing numbers of tests by disease category and facility, significant correlation was noted in toxicology (p 0.001). CONCLUSION: This research allowed comparisons in laboratory utilization between a private and a public hospital among different age groups. Differences were noted in both volume and type of laboratory tests ordered on patients with specific diagnoses in the two facilities. Although comorbidity was not well controlled for, the study does suggest that clinical laboratories may undergo changes in utilization as our nation's population ages. PMID- 15747783 TI - Factors contributing to the retention of clinical laboratory personnel. AB - OBJECTIVE: To identify factors contributing to retention of clinical laboratory practitioners. DESIGN: A paper survey addressing retention was distributed to a potential of 4000 clinical laboratory professionals. SETTING: The survey was distributed to subjects by their laboratory manager to be completed at the worksite or home. PATIENTS OR OTHER PARTICIPANTS: 599 usable surveys were received from non-supervisory individuals employed in clinical laboratory science (CLS) for five years or more. INTERVENTIONS: Surveys were mailed to laboratory managers in March 2003 with directions to distribute to practitioners with five or more years of work experience. MAIN OUTCOME MEASURES: Percentages of respondents agreeing and disagreeing with Lickert-type opinion items were determined. The means, ranges, and standard deviations were calculated for the number of hours of continuing education, years of experience, percentage of time spent on tasks, and years in the current job. The means for job satisfaction were calculated and compared statistically based on respondents' job function, satisfaction with salary, job independence, sense of appreciation, and responsibility for continuing education. Open-ended responses were tabulated and categorized. RESULTS: Committed practitioners believe their work is important and find it challenging. Those who are most satisfied with their jobs believe they make a good salary (p = 0.000), have work independence (p = 0.000), and feel that their work is appreciated (p = 0.000). Job satisfaction does not differ for CLTs vs. CLSs. Salaries comparable to nurses and appreciation from physicians, nurses, and hospital administrators are cited by respondents as the most important factors to retaining laboratory staff. CONCLUSION: Committed practitioners believe that salaries comparable to nurses are needed to improve retention of staff. Respondents said that being appreciated by hospital administrators, nurses, and physicians would also contribute to improved retention. PMID- 15747785 TI - Chronic myelocytic leukemia--Part I: History, clinical presentation, and molecular biology. AB - DATA SYNTHESIS: Chronic myelocytic leukemia (CML) was initially described in 1845 and is considered one of the first leukemias to be discovered. Diagnosis of CML was dramatically improved with the discovery of the Philadelphia chromosome by Nowell and Hungerford in 1960. However, the rudiments of our understanding of the molecular cause of CML began in 1973 when Janet Rowley discovered that the Philadelphia chromosome is a reciprocal translocation between chromosomes 9 and 22. The leukemogenic mechanisms of CML were hypothesized 20 years later when it was discovered that the t(9;22) translocation produced a fusion gene involving the BCR gene from chromosome 22 and the ABL protooncogene from chromosome 9 [corrected] Multiple breakpoints in BCR produce fusion genes that are translated into chimeric protein products of different lengths that are associated with different leukemic subtypes. CONCLUSION: Although CML has a rich history of interest to hematologists, it also represents a leukemogenic paradigm to the molecular biologist. Nearly all malignancies result from a series of mutagenic events, which culminate in full malignant transformation. However, it appears that CML results from a single mutagenic event involving the t(9;22) translocation leading to the development of the BCR/ABL fusion gene and the corresponding fusion protein. The successful transcription and translation of the BCR/ABL fusion protein led researchers to carefully study its involvement in leukemogenesis. The BCR/ABL fusion protein exhibits increased and constitutive tyrosine kinase activity that differs depending on which BCR breakpoint is expressed, resulting in varying clinical presentations. PMID- 15747784 TI - Advances in understanding the biology and genetics of acute myelocytic leukemia. AB - Acute myelocytic leukemia (AML) is a malignant neoplasm of hematopoietic cells characterized by an abnormal proliferation of myeloid precursor cells, decreased rate of self-destruction and an arrest in cellular differentiation. The leukemic cells have an abnormal survival advantage. Thus, the bone marrow and peripheral blood are characterized by leukocytosis with a predominance of immature cells, primarily blasts. As the immature cells accumulate in the bone marrow, they replace the normal myelocytic cells, megakaryocytes, and erythrocytic cells. This leads to a loss of normal bone marrow function and associated complications of bleeding, anemia, and infection. The incidence of AML increases with age, peaking in the sixth decade of life. In the United States, there are about 10,000 new cases of AML and 7,000 deaths in those with an AML diagnosis per year. Current molecular studies of AML demonstrate that it is a heterogeneous disorder of the myeloid cell lineage. This paper will discuss the most recent understanding and research of the cellular origin of AML and associated common genetic mutations that fuel the neoplastic process. Also discussed are how these advances have impacted the classification, selection of therapy, and definition of complete remission in AML. Promyelocytic leukemia will be discussed in detail as this AML subtype reveals how our understanding of the biology and genetics of the disease has led to targeted therapy that results in a cure in up to 80% of patients. PMID- 15747786 TI - Chronic myelocytic leukemia--Part II: Approaches to and molecular monitoring of therapy. AB - DATA SYNTHESIS: Chronic myelocytic leukemia (CML) was initially described in 1845 and is considered one of the first leukemias discovered. Effective approaches to therapy were not instituted until arsenic was first administered in 1865. Since then, four major therapeutic milestones have been achieved; the development of alkylating agents like busulphan and 6-thioguanine in 1953, alpha interferon in 1983, bone marrow transplantation in 1986, and tyrosine kinase inhibitors in 1998. The discovery that the protein product of this fusion gene expresses constitutive tyrosine kinase activity prompted the synthesis of a designer drug, imatinib mesylate, which binds the fusion protein and neutralizes the tyrosine kinase activity. Molecular methods of detecting BCR-ABL transcripts are showing promise in confirming drug resistance and predicting patient outcomes in response to imatinib mesylate therapy. Evidence of drug resistance can guide physicians in selecting alternative therapeutic approaches early in the course of the disease to potentially rescue non responders. Although the success of clinical trials has been dramatic, drug resistance and disease relapse are issues to be considered. CONCLUSION: The discovery that the BCR/ABL fusion protein exhibits increased and constitutive tyrosine kinase activity led investigators to develop an inhibitor to this activity. The synthesis of imatinib mesylate, currently marketed as Gleevec(TM) or Glivec(R), is in stage III clinical trials and has proven to be the most successful antileukemic drug to date. As in CML, an understanding of the leukemogenic mechanisms involved in other leukemias will provide the groundwork for the development of therapeutic interventions tailored to the specific molecular defects identified, eventually rendering obsolete the shotgun approaches to massive cell killing produced by chemotherapy. PMID- 15747788 TI - Locally linear discriminant analysis for multimodally distributed classes for face recognition with a single model image. AB - We present a novel method of nonlinear discriminant analysis involving a set of locally linear transformations called "Locally Linear Discriminant Analysis (LLDA)." The underlying idea is that global nonlinear data structures are locally linear and local structures can be linearly aligned. Input vectors are projected into each local feature space by linear transformations found to yield locally linearly transformed classes that maximize the between-class covariance while minimizing the within-class covariance. In face recognition, linear discriminant analysis (LDA) has been widely adopted owing to its efficiency, but it does not capture nonlinear manifolds of faces which exhibit pose variations. Conventional nonlinear classification methods based on kernels such as generalized discriminant analysis (GDA) and support vector machine (SVM) have been developed to overcome the shortcomings of the linear method, but they have the drawback of high computational cost of classification and overfitting. Our method is for multiclass nonlinear discrimination and it is computationally highly efficient as compared to GDA. The method does not suffer from overfitting by virtue of the linear base structure of the solution. A novel gradient-based learning algorithm is proposed for finding the optimal set of local linear bases. The optimization does not exhibit a local-maxima problem. The transformation functions facilitate robust face recognition in a low-dimensional subspace, under pose variations, using a single model image. The classification results are given for both synthetic and real face data. PMID- 15747787 TI - Automatic analysis of multimodal group actions in meetings. AB - This paper investigates the recognition of group actions in meetings. A framework is employed in which group actions result from the interactions of the individual participants. The group actions are modeled using different HMM-based approaches, where the observations are provided by a set of audiovisual features monitoring the actions of individuals. Experiments demonstrate the importance of taking interactions into account in modeling the group actions. It is also shown that the visual modality contains useful information, even for predominantly audio based events, motivating a multimodal approach to meeting analysis. PMID- 15747789 TI - Face recognition using laplacianfaces. AB - We propose an appearance-based face recognition method called the Laplacianface approach. By using Locality Preserving Projections (LPP), the face images are mapped into a face subspace for analysis. Different from Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) which effectively see only the Euclidean structure of face space, LPP finds an embedding that preserves local information, and obtains a face subspace that best detects the essential face manifold structure. The Laplacianfaces are the optimal linear approximations to the eigenfunctions of the Laplace Beltrami operator on the face manifold. In this way, the unwanted variations resulting from changes in lighting, facial expression, and pose may be eliminated or reduced. Theoretical analysis shows that PCA, LDA, and LPP can be obtained from different graph models. We compare the proposed Laplacianface approach with Eigenface and Fisherface methods on three different face data sets. Experimental results suggest that the proposed Laplacianface approach provides a better representation and achieves lower error rates in face recognition. PMID- 15747790 TI - Automated processing of shoeprint images based on the Fourier transform for use in forensic science. AB - The development of a system for automatically sorting a database of shoeprint images based on the outsole pattern in response to a reference shoeprint image is presented. The database images are sorted so that those from the same pattern group as the reference shoeprint are likely to be at the start of the list. A database of 476 complete shoeprint images belonging to 140 pattern groups was established with each group containing two or more examples. A panel of human observers performed the grouping of the images into pattern categories. Tests of the system using the database showed that the first-ranked database image belongs to the same pattern category as the reference image 65 percent of the time and that a correct match appears within the first 5 percent of the sorted images 87 percent of the time. The system has translational and rotational invariance so that the spatial positioning of the reference shoeprint images does not have to correspond with the spatial positioning of the shoeprint images of the database. The performance of the system for matching partial-prints was also determined. PMID- 15747791 TI - Real-time gesture recognition by learning and selective control of visual interest points. AB - For the real-time recognition of unspecified gestures by an arbitrary person, a comprehensive framework is presented that addresses two important problems in gesture recognition systems: selective attention and processing frame rate. To address the first problem, we propose the Quadruple Visual Interest Point Strategy. No assumptions are made with regard to scale or rotation of visual features, which are computed from dynamically changing regions of interest in a given image sequence. In this paper, each of the visual features is referred to as a visual interest point, to which a probability density function is assigned, and the selection is carried out. To address the second problem, we developed a selective control method to equip the recognition system with self-load monitoring and controlling functionality. Through evaluation experiments, we show that our approach provides robust recognition with respect to such factors as type of clothing, type of gesture, extent of motion trajectories, and individual differences in motion characteristics. In order to indicate the real-time performance and utility aspects of our approach, a gesture video system is developed that demonstrates full video-rate interaction with displayed image objects. PMID- 15747792 TI - Graph edit distance from spectral seriation. AB - This paper is concerned with computing graph edit distance. One of the criticisms that can be leveled at existing methods for computing graph edit distance is that they lack some of the formality and rigor of the computation of string edit distance. Hence, our aim is to convert graphs to string sequences so that string matching techniques can be used. To do this, we use a graph spectral seriation method to convert the adjacency matrix into a string or sequence order. We show how the serial ordering can be established using the leading eigenvector of the graph adjacency matrix. We pose the problem of graph-matching as a maximum a posteriori probability (MAP) alignment of the seriation sequences for pairs of graphs. This treatment leads to an expression in which the edit cost is the negative logarithm of the a posteriori sequence alignment probability. We compute the edit distance by finding the sequence of string edit operations which minimizes the cost of the path traversing the edit lattice. The edit costs are determined by the components of the leading eigenvectors of the adjacency matrix and by the edge densities of the graphs being matched. We demonstrate the utility of the edit distance on a number of graph clustering problems. PMID- 15747793 TI - Enhanced perceptual distance functions and indexing for image replica recognition. AB - The proliferation of digital images and the widespread distribution of digital data that has been made possible by the Internet has increased problems associated with copyright infringement on digital images. Watermarking schemes have been proposed to safeguard copyrighted images, but watermarks are vulnerable to image processing and geometric distortions and may not be very effective. Thus, the content-based detection of pirated images has become an important application. In this paper, we discuss two important aspects of such a replica detection system: distance functions for similarity measurement and scalability. We extend our previous work on perceptual distance functions, which proposed the Dynamic Partial Function (DPF), and present enhanced techniques that overcome the limitations of DPF. These techniques include the Thresholding, Sampling, and Weighting schemes. Experimental evaluations show superior performance compared to DPF and other distance functions. We then address the issue of using these perceptual distance functions to efficiently detect replicas in large image data sets. The problem of indexing is made challenging by the high-dimensionality and the nonmetric nature of the distance functions. We propose using Locality Sensitive Hashing (LSH) to index images while using the above perceptual distance functions and demonstrate good performance through empirical studies on a very large database of diverse images. PMID- 15747794 TI - Adaptively merging large-scale range data with reflectance properties. AB - In this paper, we tackle the problem of geometric and photometric modeling of large intricately shaped objects. Typical target objects we consider are cultural heritage objects. When constructing models of such objects, we are faced with several important issues that have not been addressed in the past-issues that mainly arise due to the large amount of data that has to be handled. We propose two novel approaches to efficiently handle such large amounts of data: A highly adaptive algorithm for merging range images and an adaptive nearest-neighbor search to be used with the algorithm. We construct an integrated mesh model of the target object in adaptive resolution, taking into account the geometric and/or photometric attributes associated with the range images. We use surface curvature for the geometric attributes and (laser) reflectance values for the photometric attributes. This adaptive merging framework leads to a significant reduction in the necessary amount of computational resources. Furthermore, the resulting adaptive mesh models can be of great use for applications such as texture mapping, as we will briefly demonstrate. Additionally, we propose an additional test for the k-d tree nearest-neighbor search algorithm. Our approach successfully omits back-tracking, which is controlled adaptively depending on the distance to the nearest neighbor. Since the main consumption of computational cost lies in the nearest-neighbor search, the proposed algorithm leads to a significant speed-up of the whole merging process. In this paper, we present the theories and algorithms of our approaches with pseudo code and apply them to several real objects, including large-scale cultural assets. PMID- 15747795 TI - A geometric approach to shape from defocus. AB - We introduce a novel approach to shape from defocus, i.e., the problem of inferring the three-dimensional (3D) geometry of a scene from a collection of defocused images. Typically, in shape from defocus, the task of extracting geometry also requires deblurring the given images. A common approach to bypass this task relies on approximating the scene locally by a plane parallel to the image (the so-called equifocal assumption). We show that this approximation is indeed not necessary, as one can estimate 3D geometry while avoiding deblurring without strong assumptions on the scene. Solving the problem of shape from defocus requires modeling how light interacts with the optics before reaching the imaging surface. This interaction is described by the so-called point spread function (PSF). When the form of the PSF is known, we propose an optimal method to infer 3D geometry from defocused images that involves computing orthogonal operators which are regularized via functional singular value decomposition. When the form of the PSF is unknown, we propose a simple and efficient method that first learns a set of projection operators from blurred images and then uses these operators to estimate the 3D geometry of the scene from novel blurred images. Our experiments on both real and synthetic images show that the performance of the algorithm is relatively insensitive to the form of the PSF. Our general approach is to minimize the Euclidean norm of the difference between the estimated images and the observed images. The method is geometric in that we reduce the minimization to performing projections onto linear subspaces, by using inner product structures on both infinite and finite-dimensional Hilbert spaces. Both proposed algorithms involve only simple matrix-vector multiplications which can be implemented in real-time. PMID- 15747796 TI - A quasi-dense approach to surface reconstruction from uncalibrated images. AB - This paper proposes a quasi-dense approach to 3D surface model acquisition from uncalibrated images. First, correspondence information and geometry are computed based on new quasi-dense point features that are resampled subpixel points from a disparity map. The quasi-dense approach gives more robust and accurate geometry estimations than the standard sparse approach. The robustness is measured as the success rate of full automatic geometry estimation with all involved parameters fixed. The accuracy is measured by a fast gauge-free uncertainty estimation algorithm. The quasi-dense approach also works for more largely separated images than the sparse approach, therefore, it requires fewer images for modeling. More importantly, the quasidense approach delivers a high density of reconstructed 3D points on which a surface representation can be reconstructed. This fills the gap of insufficiency of the sparse approach for surface reconstruction, essential for modeling and visualization applications. Second, surface reconstruction methods from the given quasi-dense geometry are also developed. The algorithm optimizes new unified functionals integrating both 3D quasi-dense points and 2D image information, including silhouettes. Combining both 3D data and 2D images is more robust than the existing methods using only 2D information or only 3D data. An efficient bounded regularization method is proposed to implement the surface evolution by level-set methods. Its properties are discussed and proven for some cases. As a whole, a complete automatic and practical system of 3D modeling from raw images captured by hand-held cameras to surface representation is proposed. Extensive experiments demonstrate the superior performance of the quasi-dense approach with respect to the standard sparse approach in robustness, accuracy, and applicability. PMID- 15747797 TI - Robust estimation of adaptive tensors of curvature by tensor voting. AB - Although curvature estimation from a given mesh or regularly sampled point set is a well-studied problem, it is still challenging when the input consists of a cloud of unstructured points corrupted by misalignment error and outlier noise. Such input is ubiquitous in computer vision. In this paper, we propose a three pass tensor voting algorithm to robustly estimate curvature tensors, from which accurate principal curvatures and directions can be calculated. Our quantitative estimation is an improvement over the previous two-pass algorithm, where only qualitative curvature estimation (sign of Gaussian curvature) is performed. To overcome misalignment errors, our improved method automatically corrects input point locations at subvoxel precision, which also rejects outliers that are uncorrectable. To adapt to different scales locally, we define the RadiusHit of a curvature tensor to quantify estimation accuracy and applicability. Our curvature estimation algorithm has been proven with detailed quantitative experiments, performing better in a variety of standard error metrics (percentage error in curvature magnitudes, absolute angle difference in curvature direction) in the presence of a large amount of misalignment noise. PMID- 15747798 TI - Large-scale evaluation of multimodal biometric authentication using state-of-the art systems. AB - We examine the performance of multimodal biometric authentication systems using state-of-the-art Commercial Off-the-Shelf (COTS) fingerprint and face biometric systems on a population approaching 1,000 individuals. The majority of prior studies of multimodal biometrics have been limited to relatively low accuracy non COTS systems and populations of a few hundred users. Our work is the first to demonstrate that multimodal fingerprint and face biometric systems can achieve significant accuracy gains over either biometric alone, even when using highly accurate COTS systems on a relatively large-scale population. In addition to examining well-known multimodal methods, we introduce new methods of normalization and fusion that further improve the accuracy. PMID- 15747799 TI - On using prototype reduction schemes and classifier fusion strategies to optimize kernel-based nonlinear subspace methods. AB - In Kernel-based Nonlinear Subspace (KNS) methods, the length of the projections onto the principal component directions in the feature space, is computed using a kernel matrix, K, whose dimension is equivalent to the number of sample data points. Clearly this is problematic, especially, for large data sets. In this paper, we solve this problem by subdividing the data into smaller subsets, and utilizing a Prototype Reduction Scheme (PRS) as a preprocessing module, to yield more refined representative prototypes. Thereafter, a Classifier Fusion Strategy (CFS) is invoked as a postprocessing module, to combine the individual KNS classification results to derive a consensus decision. Essentially, the PRS is used to yield computational advantage, and the CFS, in turn, is used to compensate for the decreased efficiency caused by the data set division. Our experimental results demonstrate that the proposed mechanism significantly reduces the prototype extraction time as well as the computation time without sacrificing the classification accuracy. The results especially demonstrate a significant computational advantage for large data sets within a parallel processing philosophy. PMID- 15747800 TI - An improved cluster labeling method for support vector clustering. AB - The support vector clustering (SVC) algorithm is a recently emerged unsupervised learning method inspired by support vector machines. One key step involved in the SVC algorithm is the cluster assignment of each data point. A new cluster labeling method for SVC is developed based on some invariant topological properties of a trained kernel radius function. Benchmark results show that the proposed method outperforms previously reported labeling techniques. PMID- 15747801 TI - A technique for finding the symmetry axes of implicit polynomial curves under perspective projection. AB - This paper presents an algebraic technique for detecting the symmetry axes of a perspectively projected plane curve. The procedure applies to implicit polynomials which has been fitted to 2D image data acquired by means of a photo camera or a TV set. The effectiveness of the proposed method has been demonstrated experimentally by using both synthetic and real images. PMID- 15747802 TI - Mean shift is a bound optimization. AB - We build on the current understanding of mean shift as an optimization procedure. We demonstrate that, in the case of piecewise constant kernels, mean shift is equivalent to Newton's method. Further, we prove that, for all kernels, the mean shift procedure is a quadratic bound maximization. PMID- 15747804 TI - The effect of consanguineous marriage on reading disability in the Arab community. AB - The present study examined the effect of consanguineous marriage in the Arab community on reading disabilities of offspring. It examined whether the rate of reading disabilities was higher among offspring of first-cousin parents than offspring of unrelated parents; and whether reading-disabled children of first cousin parents were more disabled in phonological awareness and phonological decoding than reading-disabled children of unrelated parents and normally reading younger children. These questions were investigated among 814 pupils of the 4th, 5th, and 6th grades, using word recognition and reading comprehension tests. Two experimental groups were chosen from this population. These were a reading disabled group of 22 pupils who were children of first-cousin marriages and 21 pupils who were children of unrelated parents. A control group was also selected, consisting of 21 younger normally reading pupils at the same reading level. All the groups were tested on non-words, real words, phonological, orthographic and working memory measures. The results indicated that the rate of reading disabilities among children of first-cousin parents was higher than that of with children of second-cousin parents, distantly related parents, or unrelated parents. Further, no differences were found in phonological awareness and decoding between the two reading-disabled groups. Moreover, the results indicate a significant advantage of the younger normal readers over the reading-disabled children in the measures of phonological awareness, decoding, and orthographical knowledge that requires spelling. However, in reading common words and choosing words in context, the performance of the reading-disabled groups and the normally reading group were similar. It has been suggested that further research is needed to evaluate the role of intelligence, nevertheless our results provide new evidence for a genetic basis to reading disabilities. PMID- 15747805 TI - A computerized treatment of dyslexia: benefits from treating lexico-phonological processing problems. AB - Two hundred sixty-seven 10- to 14-year-old Dutch children with dyslexia were randomly assigned to one of two samples that received a treatment for reading and spelling difficulties. The treatment was computer-based and focused on learning to recognise and use the phonological and morphological structure of Dutch words. The inferential algorithmic basis of the program ensured that the instruction was highly structured. The present study examined the reliability of the effects of the treatment, and provided an evaluation of the attained levels of reading and spelling by relating them to normal levels. Both samples revealed large, generalized treatment effects on reading accuracy, reading rate, and spelling skills. Following the treatment, participants attained an average level of reading accuracy and spelling. The attained level of reading rate was comparable to the lower bound of the average range. PMID- 15747803 TI - Three-dimensional scene flow. AB - Just as optical flow is the two-dimensional motion of points in an image, scene flow is the three-dimensional motion of points in the world. The fundamental difficulty with optical flow is that only the normal flow can be computed directly from the image measurements, without some form of smoothing or regularization. In this paper, we begin by showing that the same fundamental limitation applies to scene flow; however, many cameras are used to image the scene. There are then two choices when computing scene flow: 1) perform the regularization in the images or 2) perform the regularization on the surface of the object in the scene. In this paper, we choose to compute scene flow using regularization in the images. We describe three algorithms, the first two for computing scene flow from optical flows and the third for constraining scene tructure from the inconsistencies in multiple optical flows. PMID- 15747806 TI - Dyslexia in English as a second language. AB - This study focused on English as L2 in a group of Norwegian dyslexic 12 year olds, compared to an age and gender matched control group. Norwegian school children learn English from the first grades on. The subjects were assessed with a test battery of verbal and written tasks. First, they were given a comprehension task; second, a model sentence task; third, two pragmatic tasks, and fourth, three tasks of literacy. The verbal tasks were scored according to comprehension, morphology, syntax and semantics, while the literacy tasks were scored by spelling, translation and reading skills. It was hypothesized that the results of the control group and the dyslexia group would differ on all tasks, but that subgrouping the dyslexia group by comprehension skills would show heterogeneity within the dyslexia group. The data analyses confirmed these hypotheses. Significant differences were seen between the dyslexia group and the control group. However, the subgrouping revealed minor differences between the control group and the subgroup with good comprehension skills, and major differences between the control group and the subgroup with poor comprehension skills. Especially morphology and spelling were difficult for the dyslexia group. The results were tentatively discussed within the framework of biological and cognitive models of how to interpret L2 performance in dyslexia, underlining the importance of further research in L2 acquisition in dyslexia. PMID- 15747807 TI - An evaluation of a visual biofeedback intervention in dyslexic adults. AB - A prototype of a biofeedback system designed to treat dyslexia by improving heart rate variability was evaluated in a single blind study of dyslexic adults. Treatment consisted of four 15 minute exposures to a visual display synchronized with either the participant's own cardiac cycle (intervention condition), or of a synthesized cardiac cycle (placebo condition). Repeated measures were made of picture naming speed, single word reading speed and accuracy, copying speed, heart-rate variability and performance on a lateralized visual temporal order judgement task. Small but significant improvements were found in reading and naming speed in the treatment group relative to the placebo group. No significant improvements were found in unspeeded reading measures. Results from heart-rate measures indicated that treatment had effected a shift in the ratio between parameters reflecting the influence of the sympathetic and parasympathetic autonomic nervous systems (ANS), respectively, in favour of the parasympathetic. In the temporal order judgement task, participants who received treatment showed a reduced level of overall improvement relative to that seen in those who received placebo, coupled with evidence of a shift in visual attention from left to right hemifield in their pattern of performance. The results are interpreted as indicating that the treatment induces a shift in autonomic balance in favour of the parasympathetic ANS, and that this shift is also reflected in increased efficiency of left cerebral hemisphere circuits implicated in the perceptual motor processes required for naming and reading fluency. Conversely, it is also reflected in lower spatial awareness of peripheral visual stimuli, particularly those presented to left hemifield. PMID- 15747808 TI - Nasal airway changes due to rapid maxillary expansion timing. AB - The purpose of this study was to evaluate the effect of rapid maxillary expansion (RME) on nasal minimum cross-sectional area (MCA) using acoustic rhinometry (AR) in two groups of subjects who were treated before and after the pubertal growth spurt. The sample consisted of 29 patients with maxillary constriction and a control sample of 15 subjects. Both samples were divided into two groups according to individual skeletal maturation as assessed by the cervical vertebral maturation (CVM) method. Group I T (early-treated) consisted of 16 patients (eight girls and eight boys). Group I C (early-control) consisted of eight patients, and both groups had not reached the pubertal peak (CVM Stage 1-3). Group II T (late-treated) consisted of 13 patients (eight girls and five boys). Group II C (late-control) consisted of seven patients, and both groups were at a stage during or after the pubertal peak (CVM Stage 4-6). AR records were obtained for each treated subject before treatment (T1), after expansion (T2), and immediately after a three-month retention period (T3); only T1 and T3 records were obtained for controls. The overall increase in MCA was significantly greater in the early- and late-treated groups (group I T, group II T) as compared with the early and late controls. (group I C, group II C) (P < .05). The results of the present study suggest that even the overall (T1-T3) increase for MCA in group I T is greater (0.34 mm) than the increase for MCA in group II T (0.19 mm), but the difference was not significant (P > .05). PMID- 15747809 TI - A prospective study of the short-term treatment effects of the acrylic-splint rapid maxillary expander combined with the lower Schwarz appliance. AB - This prospective clinical study evaluated the short-term treatment effects of acrylic-splint rapid maxillary expander in conjunction with lower Schwarz appliance (RME-Sz) therapy to the acrylic-splint rapid maxillary expansion alone (RME-only group). Pretreatment and posttreatment lateral cephalograms were analyzed for 25 RME patients and 19 RME-Sz patients. The average time between films ranged between nine and 12 months. Statistical comparisons of the treatment changes in the RME-only and RME-Sz groups were performed by means of independent sample t-tests (P < .05). The largest difference between the two groups was in lower anterior facial height (LAFH). The normally occurring increase in LAFH was not observed in the RME-only group during the treatment period, indicating that the acrylic-splint expander had a posterior "bite block effect" on the developing craniofacial complex. LAFH increased by 1.7 mm in the RME-Sz group, a value similar to that observed in untreated individuals. Slight forward displacement of the maxilla was observed when RME was used alone, and the sagittal position of the maxilla remained unchanged in the RME-Sz group. A significant amount of intrusion of the maxillary molars (-0.8 mm) was noted in the RME-only group, whereas the maxillary molars were prevented from erupting in the RME-Sz group. The lower dentition showed a significantly greater amount of extrusion in the RME Sz group than in the RME-only group. Finally, the Sz appliance prevented the mesial movement of the lower molars during the treatment period. PMID- 15747811 TI - Efficiency of three mandibular anchorage forms in Herbst treatment: a cephalometric investigation. AB - The aim of this study was to analyze the efficiency of the mandibular banded and cast splint anchorage forms used in Herbst treatment. The records of 16 Class II:1 subjects treated with mandibular banded premolar anchorage (group A), 19 Class II:1 subjects treated with mandibular banded premolar-molar anchorage (group B), 34 Class II:1 subjects (group C), and 18 Class 11:2 subjects treated with mandibular cast splint anchorage (group D) were screened. The subjects in all four groups were treated for an average period of 0.6 years. Lateral head films were analyzed on four occasions ie, before treatment, after treatment, two years after, and four years after treatment. Mandibular anchorage loss during treatment was measured by the amount of anterior movement and proclination of the lower incisors as well as the anterior movement of the lower molars. During the treatment period a pronounced (P < .001) anchorage loss was found for all anchorage forms ie, incisor anterior movement/proclination as well as anterior movement of the lower molars. When comparing the cast splint anchorage in the Class II:1 and Class II: 2 cases, no difference was found with respect to the amount of mandibular anchorage loss of the incisors. After Herbst treatment, relapsing tooth movements were found in all three anchorage forms. It can be concluded that none of the three mandibular anchorage forms used in Herbst treatment could prevent an anchorage loss. Against all expectations, the cast splint anchorage was not better than the two banded anchorage forms. PMID- 15747810 TI - Obstructive sleep apnea patients with the oral appliance experience pharyngeal size and shape changes in three dimensions. AB - Pharyngeal size and shape differences between pre- and posttrials of a mandible protruding oral appliance were investigated using cine computerized tomography (CT). Fourteen patients diagnosed with obstructive sleep apnea whose apnea hypopnea index was higher than 5 and arousal index higher than 20 underwent a second overnight sleep study to evaluate the effectiveness of the oral appliance. Three-dimensional changes in pharyngeal shape measured on cross-sectional CT images during two respiratory cycles after oral appliance delivery were estimated by three variables: (1) lateral dimension, (2) anterior-posterior dimension, and (3) cross-sectional area at five vertical levels. Apnea indices improved significantly when the appliance was used. During apnea, measurements at retropalatal and retroglossal levels decreased most. However, the cross-sectional area of these levels appeared to increase significantly (P < .05) with the appliance in place during wakefulness. The oral appliance appears to enlarge the pharynx to a greater degree in the lateral than in the sagittal plane at the retropalatal and retroglossal levels of the pharynx, suggesting a mechanism for the effectiveness of oral appliances that protrude the mandible. PMID- 15747812 TI - Early predictive variables for upper canine impaction as derived from posteroanterior cephalograms. AB - The aim of this study was to identify a model of cephalometric variables for early diagnosis of the displacement of upper permanent canine and early prognosis of upper canine impaction. Postero-anterior (PA) cephalograms of 43 subjects (22 males and 21 females) with different types of malocclusions in the mixed dentition were analyzed at the time of first observation (mean age 8 years and 5 +/- 9 months). All subjects were reevaluated at the mean age of 14.3 years. At this time the sample was divided into 2 groups according to the presence of canine impaction: nonimpacted canine group (31 subjects) and impacted canine group (12 subjects). Stepwise variable selection on the measurements at the time of first observation identified 2 predictive variables on PA cephalograms, ie, the distance between the center of the canine crown and the midsagittal plane (A3cc to Cg Vertical) and the distance between the jugal process and the midsagittal plane (J to Cg Vertical). The closer the canine crowns to the midsagittal plane and the larger the posterior portion of the hemimaxilla, the higher the probability of canine impaction. Discriminant analysis assigned a classification power of 95.3% to the predictive model. On the basis of the equation generated by the multivariate statistical method, impaction of the maxillary canine for each new case at the age of 8 years can be predicted. The importance of PA cephalograms in the diagnosis and prediction of canine eruption disturbances is emphasized. PMID- 15747813 TI - Modulation of masticatory muscle activity by tongue position. AB - The purpose of this study was to test whether the tongue position affects the electromyographic (EMG) activities of masticatory muscles. We recorded the EMG activities of the masseter and anterior temporalis muscles in 10 skeletal Class I adults. Tongue position was monitored by two pressure transducers embedded in the midpalatal region and the lingual flange of a custom-made acrylic monoblock. We instructed subjects to assume three different tongue positions: rest, superior, and anterior. Friedman's test and Sheffe's F-test were used to statistically examine differences in muscle activities induced by changes in tongue position. Significant differences were found in masseter muscle activity between the rest and anterior positions and in anterior temporalis muscle activity between the rest and both the anterior and the superior tongue positions. We concluded that masticatory muscle activity is affected by tongue position. PMID- 15747814 TI - The association of occlusal factors with masticatory muscle tenderness in 10- to 19-year old Turkish subjects. AB - The aims of this study were (1) to investigate the relationship between occlusal factors and masticatory muscle tenderness among 10- to 19-year-old (mean 14 years eight months) Turkish subjects and (2) to identify possible sex differences between them. The sample consisted of 716 individuals (355 male and 361 female subjects). Tenderness with palpation of masseter and temporalis muscles and functional manipulation of lateral and medial pterygoid muscles was registered. The examiners recorded the Angle classification bilaterally for molars, presence of anterior and posterior crossbites, excessive overjet, open and deep bites, functional shift, and severity of anterior crowding. Associations between the occlusal factors and muscle tenderness according to sex were evaluated with chi square analysis. Statistically significant associations were found between masticatory muscle tenderness and all the investigated occlusal factors except posterior crossbite and functional shift. Masseter, medial, and lateral pterygoid muscle tenderness was higher in female subjects. Medial and lateral pterygoid muscle tenderness in Class I cases and masseter and medial pterygoid muscle tenderness in Class II, division 1 malocclusion cases were higher in female subjects (P < .05). In open-bite cases, medial pterygoid muscle tenderness (P < .05), in deep-bite cases, masseter (P < .01) and medial pterygoid (P < .05) muscle tenderness, and in excessive overjet cases, masseter muscle tenderness (P < .05) were also higher in female subjects. These results suggest that greater masticatory muscle tenderness in female subjects may contribute to the greater prevalence of temporomandibular disorders in them. PMID- 15747816 TI - Condylar disc relationships and vibration energy in asymptomatic class I 9- to 12 year olds. AB - A proper diagnosis is the key to successful treatment. The purpose of this study was to evaluate the temporomandibular joints (TMJ) by means of lateral and anteroposterior cephalograms, transcranial temporomandibular radiographs, magnetic resonance imaging (MRI), and TMJ electrovibratography (EVG) in asymptomatic and orthodontically untreated Angle Class I subjects. The study sample comprised 31 (13 boys, 18 girls) asymptomatic Class I children with a mean age of 9.7 years. The lateral cephalometric findings revealed that the subjects were mesofacial with an Angle Class I skeletal relationship, and the anteroposterior cephalograms showed a symmetrical craniofacial skeleton. Transcranial TMJ radiographs showed that the TMJs on both sides were symmetrically positioned. The time-frequency distributions of sounds from both right and left TMJs showed a wide range, and the vibrations measured by EVG were considerable. The MRI revealed unilateral disc displacement with reduction in four of the 31 subjects (13%), bilateral disc displacement with reduction in three subjects (10%), and bilateral disc displacement without reduction in one subject (3%). The data confirm that a standardized clinical examination to determine the status of the joint is not an efficient tool. This study suggests that the clinical diagnosis should be supported by extensive TMJ evaluation techniques. PMID- 15747815 TI - A longitudinal evaluation of the skeletal profile of treated and untreated skeletal Class II individuals. AB - This study evaluated the changes in the skeletal profile of Class II subjects submitted to orthodontic treatment. The experimental group comprised the lateral cephalograms of 30 Brazilian subjects (17 female and 13 male subjects) obtained between the mean ages of 11.1 years (initial) and 15.1 years (final) and treated with cervical headgear and edgewise appliance. The control group comprised the lateral cephalograms of 30 Canadian individuals (13 females and 17 male individuals) at the ages of 6, 9, 12, 14, and 16 years from the Burlington Growth Study University of Toronto, Canada, who did not receive any kind of orthodontic treatment. The results demonstrated a reduction in the convexity of the skeletal profile of both groups. However, this change was only significant for the Canadian sample of 6- to 9-year olds (P < .01), whereas reduction was greater for the Brazilian group and was significant between the initial and final ages (P < .01). In the Canadian control group, the maxilla presented a tendency toward a forward displacement (P < .01), which was not observed in the experimental group (P < .01). The mandible presented a forward displacement in both groups; yet, only the Canadian group demonstrated a significant difference, which occurred between 9 and 16 years (P < or = .01). PMID- 15747817 TI - Cephalometric features of Filipinos with Angle Class I occlusion according to the Munich analysis. AB - The purpose of this study was (1) to establish cephalometric norms for soft tissue, skeletal and dental relationships among Filipino adults; and (2) to compare these norms with the accepted German standards. Eighty-one Filipino subjects, 44 men and 37 women, were selected from the student population of the Manila Central University on the basis of the following criteria: (1) natural born ethnic Filipino, traced up to their great-grandparent's generation; (2) good facial aesthetics; (3) Angle Class I occlusion with no crowding; (4) all teeth present (third molars may or may not be present); and (5) no previous history of orthodontic treatment. Clinical examinations and interviews were conducted to ensure that the established criteria were observed properly. The German subjects, 78 men and 123 women, were selected from Hamburg and Munich on the basis of the same criteria. Each lateral headfilm was traced and digitized, and differences between the cephalometric measurements of the two groups were compared using the Student's t-test and cephalometric superimpositions. Significant differences between the two groups were seen predominantly in the lower third of the face. The Filipinos showed more posteriorly inclined apical bases and a less prominent chin. Dentally, they exhibited a bimaxillary dental protrusion resulting in an acute dental pattern and a convexity of the soft tissue profile. These findings suggest that ethnic differences in facial traits exist and awareness of the dentofacial pattern of each ethnic group will ensure better success of treatment in establishing optimal facial harmony. PMID- 15747818 TI - Changes in dentoalveolar and facial heights during early and late growth periods: a longitudinal study. AB - This longitudinal study examined the dentoalveolar structures during early and late growth periods in 62 subjects (26 male, 36 female) with normal facial patterns and acceptable occlusions. Hand-wrist radiographs were assessed and subjects grouped accordingly as either early stage (group 1) or late stage (group 2). Group 1 comprised 30 subjects with skeletal development maturation stages between PP2= and MP3=, H or S. Group 2 comprised 32 subjects with skeletal development maturation stages between MP3u and Ru. Skeletal and dentoalveolar measurements and ratios were assessed at the beginning and end of the observation periods. Differences in measurements and ratios within each group were examined using paired t-tests. Differences between the 2 groups were evaluated using Student's t-tests. Increases in alveolar heights (except for upper posterior alveolar heights) differed between the groups. Vertical alveolar dimensions increased substantially in group 1 in comparison with group 2. With the exception of upper anterior alveolar heights, all increases in group 2 were statistically significant. As a result of differential growth in alveolar dimensions, lower facial ratio (Co-Go/ANS-Me) and overbite remained constant in both stages. Vertical alveolar growth exhibited regional differentiation according to the pubertal growth periods. Although this growth plays a role in the establishment of normal facial patterns and occlusion, it should also be considered with respect to the treatment and treatment stability of patients showing vertical facial discrepancies. PMID- 15747819 TI - Frictional evaluations of dental typodont models using four self-ligating designs and a conventional design. AB - Abstract: After a previous study using typodonts and three standardized archwire (AW) sizes, the frictional evaluations of four self-ligating brackets were directed toward the optimal AW-bracket system. Four participating manufacturers suggested three AWs, which were a representation of the three stages of orthodontic treatment, to be coupled with their respective self-ligating design. Four replicated typodont models were mounted with a self-ligating design, and a fifth model was mounted with a conventional design that served as a control. The first experiment evaluated the manufacturer-suggested AWs against the respective self-ligating design. Because no third-stage AWs could engage their respective designs, a second experiment was implemented to gain more detailed analyses of the designs. This experiment included any successful manufacturer-suggested AWs from the first experiment against the four self-ligating designs and the control design. All self-ligating designs performed with the efficiency and reproducibility associated with expectations. Specifically, self-ligation outperformed the conventional brackets when coupled with up to 0.020- x 0.020 inch wires. The clearance of the various AW sizes and alloys changed with malocclusion. Furthermore, the parameter that best correlated with drawing forces was the bending stiffness of the AW, which was directly associated with the nominal dimension of each wire. The best AW-bracket system can be selected, when taking into account the stiffness (elastic modulus and size of the AW) along with the amount of malocclusion present, once the treatment plan is determined. PMID- 15747820 TI - Patient discomfort: a comparison between lingual and labial fixed appliances. AB - This study was undertaken to determine the discomfort differences between patients treated with lingual and labial orthodontic brackets. The study sample consisted of two groups of 30 adolescent patients. Group LI was treated with lingual appliances, and group LA was treated with labial appliances. After three months of treatment, each patient completed a seven-part survey with 12 questions, evaluating intraoral discomfort; tongue-lip-cheek soreness; eating, speech, and oral care difficulties; adaptation period; and general problems. In the LI group, tongue soreness and speech difficulties were significantly greater (P < .001) than in the LA group, whereas cheek (P < .001) and lip (P < .05) soreness were greater in group LA than in group LI. No statistically significant differences were found between the groups in reported intraoral discomfort and eating and oral care difficulties, but adhering of food particles was greater (P < .05) in the LI group. In this study, speech difficulty was the most severe problem for the lingual group. All patients in the LA group and 76.7% of the patients in the LI group reported that the problem was solved at the end of 30 days. However, 23.3% of the LI group claimed that at the end of three months they were still having a problem while speaking. Even the maximum adaptation period was longer in the LI group (90 days) than in the LA group (30 days). General evaluation of this study suggested that after the initial discomfort period, only a small percentage (10%) of lingual orthodontic patients reported some hindrance because of their treatment. PMID- 15747821 TI - Comparison of metal release from new and recycled bracket-archwire combinations. AB - Most orthodontic brackets and archwires made from stainless steel or nickel (Ni) titanium (Ti) alloys can release metal ions into the saliva. This study measures the amounts of metal released from simulated fixed orthodontic appliances. Sixty simulated fixed orthodontic appliances were manufactured in different ways and divided into four equal groups as follows: new brackets and new archwires (group 1 controls), new brackets and recycled archwires (group 2), recycled brackets and new archwires (group 3), and recycled brackets and recycled archwires (group 4). The bracket bases coated with adhesives were heated at 350 degrees C for 30 minutes. Recycled archwires were sterilized by autoclaving. Each appliance was soaked in artificial saliva of pH 7 at 37 degrees C for 45 days. At the end of this period, a sample of the artificial saliva was collected for analysis. Concentrations of Ni, chromium (Cr), iron (Fe), manganese, copper (Cu), and Ti ions were measured by atomic absorption and recorded in micrograms per liter. Analysis of variance and the Duncan multiple-range test indicated significant differences among the groups (P < .001). The study revealed the following: (1) group 4 released higher amounts of Cr, Fe, and Ti than any of the other three combinations; (2) Ni release was similar in groups 1 and 2 and in groups 2, 3, and 4; (3) the amounts of Cu, Cr, and Ti ions released from groups 3 and 4 were significantly greater than the amounts released from the other two combinations. PMID- 15747822 TI - Fracture strength of ceramic bracket tie wings subjected to tension. AB - One of the most common areas of ceramic bracket fracture is within the tie-wing complex. When an archwire is ligated into position, tensile forces are placed under the tie wing. However, no study, to date, has focused specifically on the fracture resistance of the tie-wing complex. The aim of this study is to compare the tensile fracture strength of seven currently available ceramic brackets (Inspire, Fascination, Mystique, InVu, Clarity, Virage, and Luxi) as a function of bracket brand and bracket configuration, semitwin vs true-twin. Based on a power analysis of pilot data, 10 maxillary central incisor brackets per group were tested to failure with a tensile load placed directly under the distoincisal tie wing. The results ranged from a maximum mean fracture strength of 147.71 (5.87) MPa with Fascination brackets to a minimum mean fracture strength of 84.28 (7.01) MPa with Luxi brackets. The statistical analysis indicated a significant effect on fracture strength as a function of bracket brand (P < .05) and that semitwin brackets, Fascination, Mystique, and Virage, had significantly higher fracture strength than true-twin brackets, Clarity, lnVu, and Luxi (P < .05). Interestingly, the only monocrystalline bracket in the study, Inspire, could not be fractured using the investigation protocol. In fact, the steel ligature fixture wire would break before tie-wing fracture at a mean fixture failure of 198.65 MPa. PMID- 15747823 TI - Longitudinal measurements of tooth mobility during orthodontic treatment using a periotest. AB - The present study was designed to investigate the alteration of tooth mobility through orthodontic treatment using a Periotest. Eighty-three crowding cases without severe skeletal discrepancies were used as subjects. For each subject, the mobility of the central and lateral incisors on both arches (U1, U2, L1, and L2) was measured immediately before and after orthodontic treatment (T1 and T2 stages) and after retention (T3 stage) by use of a Periotest. No significant differences of all the measurements were found between the left and right sides. At T1 stage, the mean values were 10.8, 10.4, 9.3, and 7.4 for U1, U2, L1, and L2, respectively. At T2 stage, the Periotest values for all the teeth increased in comparison with those at T1 stage, and significantly decreased at T3 stage compared with those at T2 stage. Negative correlations were found between the Periotest value and age, which imply that tooth mobility decreases with age. The durations of treatment and retention had low correlation with the change of tooth mobility for all the teeth. It is suggested that the tooth mobility before treatment and after retention may be one of the useful indicators for determination of the retention period. PMID- 15747824 TI - The use of Ormocer as an alternative material for bonding orthodontic brackets. AB - As new adhesives, composite resins, and bonding techniques were introduced, orthodontists adopted some of these innovations and added them to their armamentarium. The purpose of this study was to compare the shear bond strength (SBS) of two adhesive materials; one with an organically modified ceramic matrix, Admira (Voco, Cuxhaven, Germany) and another that contains the traditional Bis GMA matrix namely Transbond XT (3M Unitek, Monrovia, Calif). The new materials have a lower wear rate and are more biocompatible than traditional composites. Forty molar teeth were randomly divided into two groups: 20 teeth bonded with the Transbond adhesive system and the other 20 teeth with the Admira bonding system. Student's t-test was used to compare the SBS of the two adhesives. Significance was predetermined at P < or = .05. The results of the t-test comparisons (t = 0.489) of the SBS indicated that there was no significant (P = .628) difference between the two adhesives tested. The mean SBS for Admira was 5.1 +/- 3.3 MPa and that for Transbond XT was 4.6 +/- 3.2 MPa. It was concluded that the new material, Ormocer, which is an organically modified ceramic restorative material can potentially have orthodontic applications if available in a more flowable paste. These new materials are more biocompatible and have lower wear rate including bonding orthodontic brackets to teeth. PMID- 15747825 TI - A comparison of the shear bond strength of a resin cement and two orthodontic resin adhesive systems. AB - The object of this study was to compare the shear bond strength and the quantity of adhesive remaining on the tooth after the debonding of brackets bonded with two light-cured orthodontic resin adhesive systems (Transbond XT and Light-Bond) and a dual-cured resin cement (RelyX Unicem). Seventy-five premolars were divided into three groups. In each group, brackets were bonded with one of the adhesives according to the manufacturer's instructions. Shear bond strength was measured using a universal test machine at a crosshead speed of one mm/min, and adhesive remnant was quantified using image analysis equipment. Our results showed that the resin cement produced significantly lower bond strength than the two orthodontic resin adhesive systems. It was also observed that the bond strength produced by Light-Bond was significantly greater than that of Transbond XT. RelyX left significantly less remnant adhesive than Transbond XT and Light-Bond. Between the two orthodontic systems, Light-Bond left significantly less adhesive on the tooth than Transbond XT. PMID- 15747827 TI - C-orthodontic microimplant for distalization of mandibular dentition in Class III correction. AB - A 16-year-old male patient with a Class III malocclusion and 2 lower missing central incisors presented for treatment. The treatment plan consisted of asymmetrically distalizing the lower dentition and regaining space for lower anterior prosthetic work. C-implants were to be used as anchorage for Class III intermaxillary elastics, and two C-orthodontic microimplants (C-implants) were placed in the interdental spaces between the upper second premolars and first molars. The particular design of the C-implant head minimized gingival irritation during the orthodontic treatment. Sliding jigs were applied on the buccal for distalization of the lower posterior teeth. The correct overbite and overjet were obtained by distalizing the entire lower dentition into its proper position with C-implant anchorage, which contributed to an improvement in facial balance. It took 15 months to treat this case. The application of this new microimplant, considerations for case selection, and the sequence of treatment are presented. PMID- 15747826 TI - Effect of time on the flexural strength of glass ionomer and composite orthodontic adhesives. AB - The purpose of this study was to compare the effects of time on the flexural strength of a resin-reinforced glass ionomer and a composite adhesive system, specifically at three time frames corresponding to the three stages of polymerization of Fuji Ortho LC. Ten rectangular specimens of each material were prepared in a metal mold (25 x 2 x 2 mm) and then stored at 37 degrees C and 100% humidity in an incubator. Six test groups were created, in which each specimen was fractured using a 3-point-bending test at a crosshead speed of 0.5 mm/min. The test results indicated that there were significant differences among the groups (P = .0001). The flexural strengths were significantly higher in the two groups (III and VI) that were fractured after seven days. This was true for both the Fuji Ortho LC (x = 77 +/- 6.1 MPa) and the Concise (x = 103.9 +/- 4.2 MPa). The flexural strength of the resin-modified glass ionomer adhesive was significantly lower than that for the composite whatever the time of fracture, 10 minutes, one hour, or seven days. The analysis of the strength-deformation curve of the group of Fuji Ortho LC, which was fractured within 10 minutes after setting (group I), showed viscoplastic behavior, whereas that of all the others groups showed elastic behavior. According to this study, clinicians must consider the mechanics of Fuji Ortho LC setting and, when this material is used, wait for at least one hour to ligate initial or repaired arch wires. PMID- 15747828 TI - Lever-arm and mini-implant system for anterior torque control during retraction in lingual orthodontic treatment. AB - Anterior torque control during retraction is difficult to achieve with lingual orthodontic treatment. This article describes the use of a lever-arm and mini implant system as absolute anchorage for controlled retraction of the anterior teeth during lingual orthodontic treatment and evaluates 2 protrusion cases treated with this system. Various clinical situations are discussed and analyzed from a biomechanical standpoint. Mini-implants are needed to control the point of force application in the posterior area and produce en masse retraction with no anchorage loss. When the length of the lever-arm is adjusted to the position of the mini-implant, the desired line of action of the retraction force with respect to the center of resistance of the anterior segment is selected. Controlled retraction of the anterior teeth was achieved with no loss of anchorage. The mini implant, in conjunction with the lever-arm, is useful not only for absolute anchorage but also for anterior torque control during retraction in lingual orthodontic treatment. PMID- 15747829 TI - Re: Kaneko K, et al. Degradation in performance of orthodontic wires caused by hydrogen absorption during short-term immersion in 2.0% acidulated phosphate fluoride solution. Angle Orthod. 2004;74: 487-495. PMID- 15747830 TI - Using photodynamic therapy to treat peri-implantitis. Interview. PMID- 15747831 TI - [Effect of nocodazole on the activation of transcription factors STAT1 and STAT3 in A431 cells]. AB - The STAT transcription factors (signal transducers and activators of transcription), STAT1 and STAT3, are involved in signal transduction from growth factors and different cytokine receptors. STAT1 and STAT3 activation mechanisms are not sufficiently investigated, but they are known to depend upon both cell type and stimulus for either of them. Recently, we have shown that nocodazole blocked EGF-induced STAT1 transport to the nucleus. Here, we have compared STAT1 and STAT3 activation in response to IFNgamma, IFNalpha and epidermal growth factor (EGF) in A431 cells. We have shown the STAT1 activation by all these agents; unlike, STAT3 was activated by EGF only. STAT1 and STAT3 activation upon EGF is blocked by both nocodazole and Src-kinase family inhibitor. STAT1 activation upon IFNgamma influence is blocked by nocodazole, but does not depend on the activity of Src-family kinases. The increased STAT3 phosphorylation results from a combined action of Src-kinase inhibitor and IFNgamma. IFNalpha induced activation of STAT1 was not inhibited by either nocodazole or Src-kinase inhibitor. Taken together, the data obtained suggest that the activation of both STAT1 and STAT3 in A431 cells is accomplished by different mechanisms. PMID- 15747832 TI - [Age dymamics of stable chromosome aberration frequency in humans with natural and pathological senescence]. AB - The age dynamics of stable chromosome aberration (SCA) frequency was analysed by fluorescent in situ hybridization (FISH) in human blood lymphocytes derived from donors, irradiated by low doses of ionizing radiation (Chernobyl clean-up workers, nuclear weapon testers, etc.) and patients with hereditary premature aging--Werner's syndrome and Hutchinson-Gilford's syndrome. It was found that the level of SCA was age-dependent and increased in irradiated persons. So, the SCA level may be really an index of a so-called "radiation senescence", and may show a real biological age of irradiated persons. The patients with Werner's syndrome demonstrate increased SCA level in blood lymphocytes, corresponding to the premature aging of the organisms. But in the case of another form of premature aging--Hutchinson--Gilford's syndrome-- no rise of SCA level was found. Some possible reasons of such results are discussed. PMID- 15747833 TI - [The ultrastructure of Brassica junceae chloroplast as a criterion of salt resistance]. AB - Two salt resistant forms of the mustard Brassica junceae were obtained. These forms demonstrated no chlorophyll defects. SR2 and SR3 salt resistant forms have constant parameters of productivity in condition of NaCl salting, in comparison with the initial plants of sort Donskaya-5. Results of a comparative ultrastructural analysis of plastids show that the salt influences the plastid ultrastructure of SR2 and SR3 lines of mustard in a lesser degree in comparison with the sort plants. The plastids of SR2 and SR3 salt resistant lines of mustard are functionally quite active under stress conditions. PMID- 15747834 TI - [Mesenchymal stem cell transplantation for myocardial reparation of rat experimental heart failure]. AB - Mesenchymal stem cells (MSC) are resident pluripotent cells of bone marrow stroma. MSC have the ability to differentiate into osteoblasts, chondroblasts and adipocytes, neurons, glia and also into cardiomyocytes. The problem of MSC use in cell therapy of various diseases and in myocardial infarction therapy is widely discussed at present. The experiments were carried out on the inbred line Wistar- Kyoto rats. Myocardial experimental infarction (EI) was induced by left descending coronary artery ligation. MSC were isolated from bone marrow, cultivated in vitro and injected into the tail vein on the day of experimental infarction operation. It was shown that the structure of injured myocardium in experimental group significantly differed from that in control group. MSC transplantation led to inflammatory process acceleration and to increased angiogenesis in the damaged myocardium; also, live cardiomyocyte layers were detected in the scar. As a result, ventricular dilatation and overload of the border zone of infarct region decreased, no features of infarction relapse were shown in the border zone. PMID- 15747835 TI - [Intracellular distribution of tyrosine-phosphorylated actin-binding proteins in A431 cells spread on different ligands]. AB - Spreading A431 cells on extracellular matrix elements fibronectin, laminin 2/4 and antibody to EGF receptor (5A9 clone) leads to tyrosine phosphorylation of actin-binding proteins, which participate in focal adhesions formation. Tyrosine phosphorylation of the proteins is retained for 1 h of cell spreading. When cells interact with ligands, focal adhesion kinase (FAK) becomes tyrosine phosphorylated, and eventually phosphorylates the target proteins. The cooperative effect of integrins and EGF receptor in FAK autophosphorylation at cell spreading on antibody to EGF receptor is discussed. PMID- 15747837 TI - [Complex evaluation of wound healing process by a deep wound rat model with implanted polychlorvinyl camera]. AB - A simultaneous study of wound proteolytic activity and morphological picture of the first stages of wound healing on rat deep wound model has been shown. The process of wound healing can be evaluated by dynamics of matrix metalloproteinase activities in wound fluid. Changes in activities of different matrix metalloproteinases correlate with different stages of healing. Implantation of polychlorvinyl camera in the wound makes it possible to obtain the volume of wound fluid sufficient for a complex evaluation of healing at the initial stages of wound process. PMID- 15747836 TI - [alpha-Actinin-4 and p65/RelA subunit of NF-kappaB transcription factor are co localized and migrate together into the nucleus in EGF-stimulated A431 cell]. AB - The NF-kappaB/Rel family of transcription factors in mammalian cells regulates inducible transcription of a large number of genes in response to diverse stimuli. Despite a great number of publications on this subject, little is known about precise NF-kappaB localization in the cytoplasm. As previously demonstrated, in normal rat fibroblast and human epidermoid carcinoma A431 cells p65/RelA subunit of NF-kappaB is co-localized in the cytoplasm with actin structures. However, the mechanism of NF-kappaB interaction with actin remains unclear. We have investigated localization of p65/RelA subunit NFkappaB and alpha actinin isoforms during cell activation by epidermal growth factor (EGF). Using confocal microscopy, we have shown that alpha-actinin-4 and p65/RelA subunit of NF-kappaB transcription factor are co-localized in A431 cells. Cell treatment with EGF leads to translocation of the proteins to membrane ruffles, and eventually to migration into the nucleus. Pretreatment of A431 cells with cytochalasin D or wortmannin prior to EGF treatment increases p65/RelA and alpha actinin-4 accumulation in nuclear extracts. Co-localization of alpha-actinin-4 with p65/RelA subunit of NF-kappaB was found in nuclei isolated from stimulated cells. These results support the notion that actin cytoskeleton reorganization and alpha-actinin-4 are involved in NF-kappaB signaling. PMID- 15747838 TI - [Transcription of c-fos gene and DNA binding activity of transcription factor AP 1 increase upon differentiation of mouse F9 teratocarcinoma cells]. AB - Retinoic acid (RA) causes differentiation of mouse F9 embryonic carcinoma cell line into primitive and parietal (with dibutiril-cAMP) endoderm. The role of AP-1 transcription factor during RA-induced differentiation was studied in F9 cell line. It was shown that differentiated cells acquired protein complexes, which are specifically bound to well characterized AP-1 32P-labeled binding sites from collagenase (Col-AP-1) and c-jun (Jun2-AP-1) promoters. These complexes contain c Fos/c-Jun with Col-AP-1 site and c-Jun/ATF-2 with Jun2-AP-1 site as revealed by supershift analysis. DNA-binding activity of these complexes is high in parietal endoderm but low-detectable in undifferentiated cells. DNA-binding activity of AP 1 transcription factor correlates with increased expression of c-fos and c-jun genes. RT-PCR analysis showed an increase in steady-state level of c-fos and c jun gene transcription at the stage of parietal endoderm (terminally differentiated F9 cells). Transcription of immediate early c-fos and c-jun genes and DNA-binding activity of c-Fos/c-Jun complex are serum dependent. The rate of c-fos and c-jun gene transcription and DNA-binding activity of c-Fos/c-Jun complex decreased in serum-starved cells, but was rapidly induced upon stimulation with serum. Undifferentiated F9 cells contain a very low level of c fos mRNA, with may be a consequence of repressive chromatin structure in promoter region. Histone deacetylase (HDAC) activity is necessary to restrict expression of specific number of genes, also HDAC inhibitors are well known inductors of differentiation and anticancer agents. Frow cytometry analysis showed a decreased rate of proliferation of F9 cells after their incubation with HDAC inhibitors, sodium butirate and trichostatin A. Also, these ihibitors induced the transcription of c-fos gene. So, we conclude that HDAC activity may be necessary to sustain a high proliferative rate of undifferentiated F9 cells. PMID- 15747839 TI - [Determination of a mechanism of the coming of death by morphological methods in strangulation mechanical asphyxia]. AB - Described in the paper are morphological data characterizing the tempo and type of thanatogenesis in strangulation asphyxia. The cerebral thanatogenesis was shown to prevail in mechanical asphyxia. The results of determination of a degree of hydration of the brain by drying its samples are described. Medullary substance was demonstrated to be hypohydrated in such death. PMID- 15747841 TI - [Dynamic morphological changes observed in the respiratory system immediately after craniocerebral trauma]. AB - Microscope and morphometry examinations of specific structural changes observed in the respiratory system in death immediately after severe craniocerebral trauma denoted profound lesions with a certain sequence of progression. The extent of histomorphological changes in the lungs is shown to be influenced by a life span of victims after inflicted trauma, i.e. by the prescription of trauma. PMID- 15747840 TI - [The specificity of healing of fractures in the skull base]. AB - On the basis of actual observations we elucidated the main processes occurring in the reparation of fractures in the skull base. The specificity of reparation was shown to be conditioned by the structure of base bones and of surrounding tissues as well as by the nature of blood supply to them. Criteria are suggested for the determination if the lesions were inflicted in life time and when. PMID- 15747842 TI - [The specificity of injuries inflicted by shots made from remade and self-made guns with silencers]. AB - Discussed in the paper is a complex forensic medical evaluation of injuries inflicted by shots made from a variety of hand guns with silencers. An experimental study denoted the specificity of close-distance entry wounds at shots made from remade or self-made guns with silencers; the quantitative and qualitative differential diagnostics was possible in such cases. PMID- 15747843 TI - [Morphofunctional reaction of human adrenals at the pulmonary variation of thanatogenesis]. AB - A comparative morphometric study of male adrenals was conducted in case pulmonary thanatogenesis. A response was registered, it was associated with a dominating weight of the left adrenal (LA) and of its cortical substance over the similar parameters of the RA. The intensity and the time of the onset of reactions in different LA morphofunctional zones were found to be different in mechanical asphyxia and in carbon-monoxide intoxication. It is suggested that the occurring changes can reflect the action mechanism and result from the embryogenesis conditioned specificity of functioning of paired organs. PMID- 15747844 TI - [The specificity of thermometry of cadavers of newborns in prescribing the time of the coming of death]. AB - It is distinctly demonstrated on the basis of 7908 measurements of rectal temperature of cadavers of newborns that such thermometry should be made at a depth of at least 5.5 cm in order to ensure an accurate prescription of the coming of death (PCD) in expertise of cadavers of newborns. An optimal method of calculating the sought results as applicable to different situations and made with the help of correction factors is described. The results of lifetime thermometry of 60 newborns are presented, which prove that the length of intrauterine life of a baby affects essentially its values. An accurate PCD of a newborn is a technical study result. PMID- 15747845 TI - [An algorithm of computerized processing of molecular-genetic data for personality identification based on the outcome of events involving numerous victims]. AB - Computerized systems of data processing are a high-efficiency tool for the storage and processing of big volumes of molecular genetic data as well as for the forensic medical identification of personality in events involving enormous unidentified bodies coming from zones of calamity or natural disaster. Described in the paper is a sequence of actions undertaken by the expert when the DNAdacto and mDNAbase soft tools are in use. The algorithm of computer-based expert analysis of typing of chromosome and mitochondrial DNA was tested at the 124th Central laboratory of medical-criminal identification of Russia's defense ministry; it was proven to be highly effective for identification made in events with numerous unidentified bodies. PMID- 15747846 TI - [ELISA-based determination of opiates in purulent transformation of cadaveric material]. AB - Abuse of drugs has been a topical issue up to the present-day. Opiates like morphine and heroin are still prevalent. The routine methods of determination of drugs in blood made within the forensic medical expertise are time-consuming and samples must be specially prepared for them. ELISA, when used for the determination of opiates in blood and urine, significantly cuts the test time. According to our research results, the diagnosis of opiates by ELISA is advisable at the preliminary stage of the forensic expertise of cadaver; the method is informative, which is of extra importance in cases of purulent transformation of cadaveric material. PMID- 15747847 TI - [Forensic medical diagnostics of nitrate and nitrite intoxication]. AB - Nitrates and nitrites, accumulating in toxic concentrations in soil, plants, water and foodstuffs, can cause lethal intoxication. The method of isolation, identification and quantitation has been elaborated for their diagnosis. It is based on the isolation of water and subsequent chromogenic test plus photoelectrocolorimetry. PMID- 15747848 TI - [Efficiency of hardware-assisted tools in forensic expertise]. AB - The authors analyze the efficiency of special-purpose hardware-assisted tools (SHT) in forensic expertise. Unlike clinical medicine, in which the methods of quantitative and qualitative analysis are comprehensively used in choosing drugs, issues related with providing the state forensic-expertise facilities with specialized equipment have not been virtually elaborated up to now. The qualitative analysis based on the subjective evaluation of experts is used presently for the purposes. Therefore, development of objective criteria of evaluation of SHT for forensic expertise is an important and topical issue. PMID- 15747849 TI - [Digital photography as an object of forensic medical examination]. AB - The possibility of diagnosing skin lesions by means of digital pictures was investigated. A "Zenit ET" film camera, HP flatbed scanner, amateur "'Panasonic" and "Sony" videocameras, miniature USB camera and digital photocameras were used to obtain images. Above 1000 images were studied. Criteria were fixed for the required quality of images. A method was developed to formalize images by means of a color etalon and mathematical assessment of formalized skin lesions. Stages were suggested for the computer-based analysis of images. Computer software was developed for the automated formalization of images and for the statistical processing of values of image elements. It was demonstrated that digital photography can be an independent informative object of forensic medical examination. PMID- 15747850 TI - [The clinical-and-morphological specificity of myocarditis with lethal outcome]. PMID- 15747852 TI - [Modern potentialities of examination of imprints on exhibits]. PMID- 15747851 TI - [Naphthalene intoxication]. PMID- 15747853 TI - [S. A. Gromov--founder of home forensic medicine]. PMID- 15747854 TI - [80 years of forensic medicine in the Maritime Territory]. PMID- 15747856 TI - Community perceptions of important camel diseases in Lapur Division of Turkana District, Kenya. AB - This paper presents the results of a study conducted in Lapur Division of Turkana District, Kenya, to estimate the incidence and mortality of camel trypanosomosis using participatory methods. Four livestock camps ('adakars') were conveniently selected for the study. Four informant groups comprising 6 8 key persons were used for the participatory exercises. The camel diseases identified by the pastoralists in their order of importance according to annual incidence were: trypanosomosis (11.4%); mange (10.8%); tick infestation (7.9%); haemorrhagic septicaemia (7.7%); and non-specific diarrhoea (7.6%). Almost half (49.3%) of the camel population suffered from at least one disease over the previous year. The annual incidence and mortality rates of trypanosomosis were estimated at 15%, and 9.9%, in adult camels and 6.9% and 5.2%, in young camels, respectively. There was a seasonal occurrence of trypanosomosis, with most cases reported in the dry season. The prevalence levels of the disease reportedly declined from about 100%, in 1978 to an almost stable state of about 15% in 2002. This study revealed that camel trypanosomosis is still an important disease in Turkana District, exacting a heavy toll in terms of morbidity and mortality. The economic losses due to the disease were likely to have been great owing to the central role the camel plays in this arid district of Kenya. PMID- 15747855 TI - Haemolytic complement activity and humoral immune responses to sheep red blood cells in indigenous chickens and in eight German Dahlem Red chicken lines with different combinations of major genes (dwarf, naked neck and frizzled) of tropical interest. AB - A total of 376 chickens from different ecotypes were immunized with the non pathogenic multi-determinant antigen sheep red blood cells (SRBC). The ecotypes included indigenous chickens from various locations in Tanzania (n=102), India (n=86) and Bolivia (n=89). In addition, eight German Dahlem Red (GDR) chicken lines with different major genes (dwarf, naked neck and frizzled) of tropical interest were also immunized with SRBC. Immune competence of the breeds was assessed by measuring complement haemolytic activity, both from the classical calcium-dependent complement pathway (CPW) and alternative calcium-independent complement pathway (APW), alongside IgTotal, IgG and IgM antibody responses to SRBC at 7 days post immunization. Large variations in complement activity and antibody responses to SRBC were observed within and between the indigenous breeds. Many indigenous chickens, especially from Bolivia, showed decreased complement activity (APW) following immunization with SRBC. Breeds from India showed the highest CPW activity and humoral (especially IgM) responses to SRBC, suggesting high immune competence. In contrast, Bolivian chickens were characterized by low CPW activity, low APW activity and low antibody levels to SRBC suggesting an overall low immune competence. In the GDR chickens, characterized by high CPW activity and high IgG antibody responses to SRBC, the major genes for naked neck, frizzling and dwarfism had no significant effect on the antibody responses and complement activity to SRBC. PMID- 15747857 TI - In vivo effects of Fagara leaves on sheep infected with gastrointestinal nematodes. AB - Infections of the gastrointestinal tract with parasitic nematodes, and particularly Haemonchus contortus, represent one of the main pathological constraints on the breeding of small ruminants in tropical countries. Owing to the constant development of anthelmintic resistance in worm populations, alternative control methods are being evaluated, including an assessment of the value of the traditional plants. The effect of the consumption of Fagara leaves on sheep infected with gastrointestinal nematodes was therefore measured both under controlled conditions and in a field survey. In sheep experimentally infected with 2500 third-stage larvae of H. contortus, a three-day administration of Fagara was associated with a decrease in egg excretion and a significant reduction in the fertility of female worms, without changes in the worm number. A reduction in egg output was also confirmed from the results obtained of a survey or naturally infected sheep performed in smallholder farms. This survey also indicated that the repeated consumption of small amounts of Fagara leaves was more effective than a single three-day administration. Results of both studies suggest that Fagara leaves might be an alternative option to chemotherapy to modulate the epidemiology of H. contortus in tropical countries. PMID- 15747858 TI - Prevalence of infection with Fasciola gigantica and its relationship to carcase and liver weights, and fluke and egg counts in slaughter cattle and buffaloes in southern Mindanao, Philippines. PMID- 15747860 TI - Growth performance of lambs in Phangrang, Vietnam: effects of a dietary supplement containing prickly-pear cactus. AB - In Phanrang, Vietnam, sheep production is limited by the sparse availability of green roughage. Although prickly-pear cactus (Opuntia elator) is abundant in the area, it is not used. It was considered relevant therefore to test whether prickly-pear cactus could be used a component of feed supplements. With the use of the cactus and other local products--namely groundnut cake, fish sauce by product, molasses, rice bran and cassava flour--two supplements were prepared. The level of cactus in the two supplements was 670 g/kg product (320 g/kg dry matter). The cactus was burned to remove the spines, chopped and sun-dried before use. The cactus preparation contained only 12 g crude protein/kg (68 g/ kg dry matter). The protein content of the two supplements was raised with either groundnut cake or fish sauce by-product. Sheep that were grazing during the day were offered either no supplement or one of the supplements when confined during the night. During a period of 3 months, non-supplemented control sheep gained 98 +/- 10.5 g/day (mean +/- SD, n=10). Sheep supplemented with cactus and groundnut gained 145 +/- 12.3 g/day, and those given cactus and fish sauce gained 130 +/- 11.7 g/day. It is concluded that prickly-pear cactus has potential as component of feed supplements for sheep. PMID- 15747859 TI - Anthelmintic activity of extracts of Spondias mombin against gastrointestinal nematodes of sheep: studies in vitro and in vivo. AB - This study was carried out to validate the efficacy of Spondias mombin, used locally as an anthelmintic, and to standardize the effective dose of the plant extract required for worm control in livestock. In vitro and in vivo studies were conducted to determine the direct anthelmintic effect of ethanolic and aqueous extracts of S. mombin towards different ovine gastrointestinal nematodes. A larval development assay (LDA) was used to investigate the in vitro effect of extracts on strongyle larvae. Another study was conducted in vivo to evaluate the therapeutic efficacy of the extracts administered orally at dose rates of 125, 250, 500 mg/kg to sheep naturally infected with gastrointestinal nematodes. Twenty sheep were selected on the basis of positive faecal egg counts (750 epg). The sheep were allocated randomly to a non-medicated control group (A) or to groups given 125 mg/kg (B), 250 mg/kg (C) or 500 mg/kg (D) of extract, respectively. Sheep in groups B-D were given extracts orally on two days. Individual faecal egg counts were performed on days 0, 3, 6, 9 and 12. The presence of S. mombin extracts in in vitro cultures of larvae decreased the survival of L3 larvae. The LC50 of the aqueous extract of S. mombin was 0.907 mg/ml, while the LC50 of the ethanolic extract was 0.456 mg/ml. This difference in LC50 was statistically significant (p > 0.05). The mean percentage faecal egg reduction of sheep drenched with 500 mg/kg S. mombin extracts was 15.0%, 27.5%, 65.0%, 65.0%, 100.0% against Haenmonchus spp., Trichostrongylus spp., Oesophagostomunm spp., Strongyloides spp. and Trichuris spp. respectively, on day 12. Extracts of S. mombin could find application in the control of helminths in livestock. PMID- 15747861 TI - Study on coccidiosis in Kombolcha Poultry Farm, Ethiopia. AB - A study on the occurrence of coccidiosis and distribution of Eimeria species in dead chickens 1-60 days of age, at Kombolcha Poultry Multiplication and Research Center (KPMRC), Ethiopia was conducted from November 2002 to April 2003. Out of the 965 dead birds, 370 (38.34%) were found to have clinical coccidiosis. The Eimeria species identified in this study were Eimeria brunetti, E. tenella, E. acervulina and E. necatrix with prevalence rates of 45.3%, 40.8%, 9.7%, and 4.1%, respectively. In the current study, E. brunetti was reported for the first time in Ethiopia. It was noted that clinical coccidioisis was more prevalent in those between 5 and 6 weeks of age. A statistically significant difference (p < 0.05) was observed in clinical coccidiosis prevalence among the different age groups studied. Various managerial problems that are associated with this high prevalence of clinical coccidiosis are identified and appropriate control strategies are recommended. PMID- 15747862 TI - Controlling health costs and improving health care quality for retirees. PMID- 15747863 TI - [Anti-measles DNA-immunization in experiment: immunogenicity and safety]. AB - The plasmid DNA pCDNA3.1-H encoding the N-terminal sequence of the measles hemagglutinin (H) protein was constructed. Virus-specific particles (VSP) containing the plasmid DNA pCDNA3.1-H coated by the spermidine-polyglucin complex. The mice were immunized by VSP. ELISA, HAIT and immunoblot showed a shaping specific humoral response. The sera of immunized animals were proven to neutralize the wild strain of the NOV96 measles virus. The formation of the specific cell immunity was confirmed by erythrocyte proliferation assay and ELISpot. PCR was used to detect the presence of the plasmid DNA in different intestines and tissues of animals after a single immunization. It was not detected at any time interval in the brain, liver, thymus and blood. And it was present on days 7 and 14 in the red bone marrow, spleen, muscular tissue, lungs and fatty tissue. On day 21 the plasmid DNA was not detected in any of the investigated organs. PMID- 15747864 TI - [Quantitative laboratory methods for the diagnosis of cytomegalovirus infection in premature infants]. AB - 31 prematures with signs of the cytomegalovirus infection (CMV) were examined. The blood and urine samples were tested for direct viral markers, i.e. for infectious CMV by the rapid culture method (RCM) and for viral DNA by quantitative PCR. Besides, the parameters of the specific immune response were studied in the babies. CMV was detected by RCM and/or PCR in 25 of the 31 examined babies during their 1st life week. The highest content of CMV within the investigated samples, i.e. 100 antigen-containing cells per 2.5 x 10(5) culture cells and above 2000 copies/ml of viral DNA was detected in 8 (32%) children. The quantity of viral DNA did not exceed 1000 copies/ml and one to three of stained cells was detected by PCR in 13 (42%) children. A study of anti-CMV in sera revealed high-titer of AT IgG in all 30 children. High avidity of anti-CMV-IgG was demonstrated to correlate with a low viral load and a low CMV infection activity in the newborns. According to the results, at least 3 laboratory diagnosis tools should be used in the diagnosis, they are PCR, RCM and determination of the anti-CMV avidity. PMID- 15747865 TI - [Detection of direct markers of cytomegalovirus. Research of the spectrum and avidity of antiviral antibodies in infants and preschool children]. AB - Thirty-three children aged 1 month to 3 years were examined within the case study. spELISA, immunoblot (IB), shell vial method (SVM) and PCR, were used for the detection of anti-CMV IgM and IgG, in the diagnosis of cytomegalovirus (CMV). Clinical signs of CMV infection (CMVI) were registered in 20 children (group 1); no CMVI specific signs were detected in the remaining 13 children (group 2). Class M antibodies were identified in 50% of group-1 sera. Around 80% of children in the group had anti-CMV-IgG. AI < 0.6 was in 3 (20%) of 15 examinees. Direct CMV markers (DNA and infection activity) were detected in 13 (65%) of 20 children. Sera of 13 children with non-specific symptomatology (group 2) had no anti-CNV-IgM, while IgG were found in 54% examinees in the group. The infectious active virus was not detected in a single baby. The used laboratory tools enhance the efficiency of CMVI diagnosis and denote a disease variation. PMID- 15747866 TI - [Gene expression of the interferon and cell-immunity systems in human blood samples]. AB - The interferon (IF) and cell-apoptosis (CA) systems are interrelated and regulate the protective reactions in body by means of a complex of biologically active proteins. The transcription levels of mRNA were determined by the RT-PCR semi quantitative method in order to compare the constitutive expression levels of IF (alpha, beta, gamma) genes, IF-dependent enzymes of 2'5' oligoadenylatesynthetyase (OAS), RNAase L, dsRNA-protein kinase (dsPK) and CA effectors (Fas-Ag, bcl-2 and gamma-actin) in human blood microsamples. cDNA dilutions, different numbers of amplification cycles (PCR with specific pairs of primers) as well as PCR-products' dilutions for dot-hybridization with specific probes were made use of to detect and evaluate levels of 9 mRNAs. The constitutive levels of gene expression of the IF and CA systems were found to differ essentially from others (1000-fold). The studied mRNA types were shared between 5 groups according to their transcription levels: very high--alpha-IF, high--RNAase L, medium--gamma-actin and bcl-2, low--beta-IF and very low (detectable after induction only)--gamma-IF, OAS, dsPK and Fas-Ag. The used detection method has a sufficiently high sensitivity and can be recommended for studies of IF inductors with unknown action mechanisms. PMID- 15747867 TI - [ELISA and RT-PCR-based research of viruses in the ticks collected in the foci of Crimean-Congo fever in Kazakhstan and Tajikistan in 2001-2002]. AB - Different species of ticks were found, in the territories of Kazakhstan and Tajikistan, to be infected with the virus of Crimean-Congo hemorrhagic fever (CKHF). The virologic evaluation included determination of antigen and RNA of the CKHF virus by ELISA and RT-PCR, respectively. The below tick species were found to be involved in the epidemic process: Hyalomma asiaticum, Dermacentor niveus (Kazakhastan) and Hyalomma anatolicum (Tajikistan). The results testify to the fact that Hyalomma ticks are the main carrier of the above virus in the Middle Asia. At the same time, Dermacentor niveus ticks are infection carriers in Kazakhstan. PMID- 15747868 TI - [Residual neurovirulence of the chimera of Langat and Denge-4 flaviviruses in intracerebral infection of monkeys]. AB - The residual neurovirulence of the previously obtained chimeric virus Tp21/DEN4, containing the RNA region encoding the pre M and E structural enzymes, strain Tp21, virus Langat, and the remaining part of the genome from the Denge 4 virus, was studied in experiments with monkeys Cercopithecus aethiops involving the intracerebral administration of the virus. The tick-borne encephalitis virus, strain Absettarov, was used as positive control. A comparative analysis of the experimental and published data showed the chimeric virus to be less virulent by its degree of morphological affection in the CNS zones, its spread into the CNS and by a percentage share of animals with viremia versus the Tp21 parent strain and Elantsev strain of the Langat virus. PMID- 15747870 TI - [Herpesvirus infection in patients with chronic glomerulonephritis]. AB - Virological examinations of blood, urine and saliva in 75 patients with chronic glomerulonephritis (CG) revealed, in 95% of them, herpes-virus infections caused by herpes simplex virus, type 1 (34.4%), herpes simplex virus, type 2 (2.6%) and cytomegalovirus (12%) or mixed infections (46%). The infection rate in the control group of children without renal pathology was reliably lower. A majority of CG patients (94%) had a diagnostically significant level of antiherpetic antibodies, class IgG, which also evidence to chronic herpes-virus infection. PMID- 15747869 TI - [Lissavirus in bats residing in the South of West Siberia]. AB - We caught, 2002-2003 in caves of Novosibirsk Region and Altai Territory, 52 bats (Myotis daubentoni, Myotis brandtii, Murina leucogaster, Plecotus auritus). Examinations of the animals' brain samples by fluorescence antibody microscopy and RT-PCR revealed lyssaviruses, genotype 1, which were close to strains provoking rabies in the local animals. According to the results, bats can be involved in keeping alive the rabies foci in Western Siberia, which must be taken into account in antiepizootic measures. PMID- 15747871 TI - [Hepatotropic action of the virus of West Nile fever]. AB - An experimental infection of mice with West Nile Virus (WNFV) showed pronounced dystrophic changes in the hepatic parenchyma and expression of WNV antigens in the endothelium of hepatic capillaries and in hepatocyte cytoplasm, which testifies to the tropic action of WNFV to hepatic tissue. PMID- 15747873 TI - [Cultivation of the vaccine strain of the virus of porcine classic plague]. AB - Several cell cultures were compared and examined for their capability to support reproduction of Classical Swine Fever Virus (CSFV). Experimental conditions of virus cultivation were optimized. The subculture of primary lamb testicular (LT) cells was selected for large cell cultivation of CSFV. In contrast to many cell lines, these cells were free of Bovine Viral Diarrhea Virus (BVDV) contamination partly due to liquid nitrogen storage and gamma-ray sterilization of serum used in the growth media. The vaccine strain CS of CSFV was obtained in the titers of 6.7-7.7 TCID50/ml under the optimal conditions of virus reproduction in the LT cells. PMID- 15747872 TI - [Optimization of RT-PCR for the identification of VP4 gene of group A rotaviruses and evaluation of its diagnostic efficiency]. AB - A method is suggested for the detection of the human rotavirus VP4 gene by RT PCR. The method is universal for all [P] types of group A rotavirus in clinical samples. As against RNA electrophoresis and ELISA, it provides for a more accurate detection of rotaviruses by 18.8 and 26.5%, respectively. It was established by RT-PCR in February, 2002, in Nizhny Novgorod, that rotaviruses were responsible for 67% of acute gastroenteritis in hospitalized children. PMID- 15747874 TI - Anatomical study of meandering and functions of human intralaryngeal artery. AB - In recent years, partial laryngectomy and partial reconstruction are increasingly intended for conservation of functions of phonation and swallowing. In partial reconstruction, it is important to comprehend morphological characteristics of the blood vessels distributed in the larynx, but there have been only few reports discussing detailed information about them. Previous reports on laryngeal blood vessels have shown that branches of some arteries show remarkable "meandering". In the present study, we devised a method for objectively determining the morphological nature, "meandering" and assessed functions of the arteries. Intralaryngeal arteries were excised from the larynx of cadavers prepared for practice in anatomy, and images of the "meandering" artery were analyzed with NIH Image. The extent of "meandering" was expressed mainly as the ratio of the total length of the blood vessel to the distance between the starting point and the end point of meandering. The results showed that there was a significant difference in the extent of meandering between superior posterior and medial posterior branches of superior laryngeal artery. These arteries, which were distributed in the arytenoid region, were found to be of primary importance in partial laryngectomy and partial reconstruction of the larynx. PMID- 15747875 TI - No raphe identified in the orbicularis oculi muscle. AB - This study was performed to elucidate whether the raphe of the orbicularis oculi muscle (raphe) exists or not. Nine upper eyelids of 6 Oriental cadavers with ages at death ranging from 72 to 91 years were dissected; 6 for gross dissections and 3 for histological slice sections. After removing the lateral half of the eyelid skin, the lateral part of the orbicularis oculi muscle and its subjacent tissue were observed macroscopically. The full layered tissue of the 8 mm lateral part from the orbital rim was incised perpendicularly and sections sliced, which were then observed microscopically after staining with the hematoxylin and eosin. The raphe was not identified macroscopically or microscopically. The lateral part of the orbicularis oculi muscle was continuous without the tendinous intercalation; under it, fibrous connective tissue corresponding to the lateral orbital thickening was observed, and in which the band configuration, microscopically the tendinous structure, was formed. The raphe was not identified. The physiological role of the lateral part of the orbicularis oculi muscle is maintained by a less tight attachment of the lateral orbital thickening, but not by the raphe. PMID- 15747876 TI - Three ligaments reinforce the lower eyelid. AB - This study was performed to elucidate the ligamentous structures in the lower eyelid other than the Lockwood ligament. Ten lower eyelids (right 5, left 5) of 5 Oriental post-mortems were dissected. The orbital septum was incised along the orbital rim, and then turned over toward the eyelid margin. The orbital fat was exposed and removed as much as possible without injuring the ligamentous structures. The ligamentous structures in the lower eyelid were then investigated macroscopically. There were two other ligaments in the lower eyelid in addition to the Lockwood ligament. All three ligaments originated from the posterior lacrimal crest, but went in different directions. The superior ligament ran on the junction of the orbital septum and the capsulopalpebral fascia, and in the lateral area, it ran on the posterior aspect of the orbital septum and attached to the periosteum of the lateral orbital rim. The inferior ligament went toward the posterior aspect of the inferior oblique muscle after separating from the Lockwood ligament. Ligaments other than the Lockwood ligament were revealed in the lower eyelid. These ligaments are thought to reinforce the fragility of the lower eyelid. PMID- 15747877 TI - Distribution of facial motoneurons innervating the common facial muscles of the rabbit and rat. AB - The distribution of the facial neurons that innervate several facial muscles was determined in the rabbit and the rat by examining the retrograde transport of horseradish peroxidase (HRP). The target muscles were musculus levator nasolabialis, m. levator labii superioris, m. zygomaticus, and m. buccinator pars buccalis, as well as m. parietoauricularis and m. depressor anguli oris in the rabbit and m. levator auricularis posterioris in the rat. Localization of the retrogradely labeled neurons within the ipsilateral facial nucleus was confirmed for all facial muscles examined. Our results showed that m. levator nasolabialis was innervated by neurons located in the dorsal subnucleus, while the motoneurons innervating m. buccinator pars buccalis were distributed within the dorsal part of the intermediate subnucleus of the facial nucleus in the both species. Localization of the labeled motoneurons innervating m. zygomaticus and m. levator labii superioris showed the difference in the distribution within the facial nucleus among the species. Neurons innervating m. parietoauricularis and m. levator auricularis posterioris were localized in somewhat different subregions of the medial subnucleus in these species. M. depressor anguli oris was innervated by the neurons distributed within the intermediate subnucleus of the facial nucleus in the rabbit. Thus, our findings revealed that there is species specific motor innervation pattern in rabbits and rats, despite several movement of the face is supplied by the homologous facial muscles. PMID- 15747878 TI - Projection from the facial nucleus to the auricular muscles of rabbits. AB - The auricular muscles correspond to the group of facial muscles that allows the auricle to move freely. The shape of the auricle and the location of the muscles on the head differs among species. The rabbit has eight auricular muscles that are innervated by motoneurons originating from the facial nucleus located on the ventral side of the medulla oblongata. However, the distribution within the facial nucleus of the motoneurons that innervate each auricular muscle has not been defined clearly. In the present study, retrograde transport of horseradish peroxidase was used to identify groups of facial motoneurons in the rabbit. Our results show that the medial subnucleus of the facial nucleus projects primarily to the auricular muscles, and that each auricular muscle is innervated by a specific group of motoneurons distributed in a particular region of the medial subnucleus. This motoneuron patterning appears to be a species-specific characteristic of the innervation of the auricular muscles. PMID- 15747879 TI - [Is it effective to make plans for the future?]. PMID- 15747880 TI - [Shift-work management in nursing]. PMID- 15747881 TI - ["Have you now changed the face?"]. PMID- 15747882 TI - ["What can I put in motion here?"]. PMID- 15747883 TI - [Health promotion in family nursing]. PMID- 15747884 TI - [Potential for optimization in nursing documentation]. PMID- 15747885 TI - [Fast Track--or the fastest possible treatment method]. PMID- 15747886 TI - [Accompanying children into their death]. PMID- 15747887 TI - [Ensuring the future--shaping the future]. PMID- 15747888 TI - [Pleading for a discussion]. PMID- 15747889 TI - [Being well prepared for a future foundation]. PMID- 15747890 TI - [The certain vacation]. PMID- 15747891 TI - Abuse in the elderly--a perennial problem. PMID- 15747892 TI - Impact of new prostheses on the oral health related quality of life of edentulous patients. AB - OBJECTIVE: A study was conducted to evaluate the impact of the placement of complete dentures by using the Global Oral Health Assessment Index (GOHAI). BACKGROUND: Oral health quality of life indicators can be used to evaluate the effects of dental treatments. MATERIAL AND METHODS: The 26 participants were treated in a French University Clinic during 2002. They were randomly divided into two groups. Each group received new prostheses, but evaluation of the quality of life was made at different periods [baseline, denture placement (group 1), 6 and 12 weeks (group 2) after placement]. A questionnaire was used to collect information on patient's satisfaction with the previous and new prostheses. Nonparametric tests were used to test the relationships between patients' satisfaction or baseline data and GOHAI variations with time as well as to compare mean values of GOHAI within each group. RESULTS: At baseline, the impact of oral health problems was apparent; the mean GOHAI-Add score was 45.8 (10.2). Six weeks after placement of the new denture, there was no difference in GOHAI scores compared with the initial assessment. An improvement in GOHAI score was observed 12 weeks after the participants received their new dentures (p < 0.05). Change in GOHAI-Add scores was negatively correlated with the initial GOHAI-Add score. Patients who preferred the new prosthesis enjoyed a positive change in GOHAI scores (p < 0.001). There was a relationship between participants' satisfaction with the new dentures and change in GOHAI scores (p < 0.05). CONCLUSION: The GOHAI can be used to evaluate needs for and effect of the making of new complete dentures. PMID- 15747893 TI - The masticatory handicap of wearing removable dentures in elderly men. AB - OBJECTIVE: This study aimed at describing the masticatory handicap of wearing removable dentures measured by some masticatory tests and the intake of hard and soft foods. MATERIALS AND METHODS: The subjects were participants in a comprehensive health examination of 67-68-year-old men living in Malmo, Sweden. Four hundred eighty-three men took part in a clinical oral examination which recorded the number of teeth and removable dentures. One hundred and one had complete dentures in both jaws and 131 had removable partial dentures in different combinations. Masticatory tests used almonds to record the number of stokes to the first swallow, and two-coloured chewing gums for recording bolus kneading and shaping. A nutritionist paid a home visit to assess dietary habits including the consumption of hard and soft foods. The data could be split into groups of different tooth and denture situations which were large enough to enable statistical analysis to be carried out. The results of a regression analysis of the group with different numbers of natural teeth (NT) and no removable dentures could be used as a reference for correlation with the masticatory capacity of removable denture wearers. RESULTS: Number of strokes to the first swallow revealed no masticatory limitations of wearing removable dentures, while chewing gum colour-mixing and shaping revealed more differentiated impairments equivalent to the function of five to 16 teeth in a remaining natural dentition. Hard food intake for the removable denture groups was comparable to 17-19 NT. Soft food intake was not influenced by denture wearing. In a ranking of oral conditions, those with more than 24 NT had the highest test values for all tests, and those with complete sets of dentures the lowest except for the number of strokes to the first swallow of an almond. CONCLUSION: Removable denture wearing can be regarded as a handicap when measured with objective masticatory tests using chewing gum and the intake of hard foods. The number of chewing strokes to the first swallow of an almond and the intake of soft foods is not affected by the wearing of removable dentures. PMID- 15747894 TI - Oral health-related well-being of the long-term hospitalised elderly. AB - OBJECTIVE: To investigate oral health-related well-being of the long-term hospitalised elderly as reported by their primary nurses in relation to subject's oral health assessed either by primary nurses or by a clinical dental examination. BACKGROUND DATA: Little is known about oral health-related well being of the medically compromised, long-term hospitalised elderly, most of whom are unable to express their feelings and opinions. MATERIALS AND METHODS: A cross sectional study using a questionnaire for primary nurses about oral health and oral health-related well-being regarding functional, pain/discomfort-related, and psychosocial limitations of the subjects (n = 255) and assessment of oral health by clinical examination. The total number of limitations and the number of limitations in each category was calculated. RESULTS: Most (77%) of our subjects were unable to eat normal food. Functional limitations dominated followed by psychosocial and pain/discomfort-related limitations. Overall assessment by each subject's primary nurse ranked oral health of as good for 9% of subjects, as moderate for 44%, and as poor for 47%. Clinical examination-based assessment ranked oral health as good for 19%, as moderate for 33%, and as poor for 48% of our subjects, with good oral health being ranked as good for more men than women (26% vs. 16%; p = 0.045). Fewer limitations were recorded for those with better oral health assessed both by primary nurse and by clinical examination. CONCLUSION: More efforts are called for to maintain the oral health of the long term hospitalised elderly so as to improve their well-being. PMID- 15747895 TI - The advantages of occupational therapy in oral hygiene measures for institutionalised elderly adults. AB - OBJECTIVE: To investigate a new method in teaching and supervising tooth and denture brushing activities by employing occupational therapy techniques. MATERIALS AND METHODS: Sixty-one residents, 44 women and 17 men, with an average age of 85.7 +/- 6.6 years (range 72-97 years) living in a Long-Term Care home (LTC) in Geneva were enrolled in a randomised controlled trial. They were divided at random into experimental (EG) and control groups (CG) with matched age and sex distribution. Two subjects passed away during the 3-month experimental period. Following medical history, plaque scores and tooth brushing habits were evaluated within the context of a comprehensive clinical assessment. Furthermore, a Mini Mental State and a vision test were taken. Based on the results of these health assessments both the EG and the CG were divided into an 'assisted' (IA) and an 'independent' (II) subgroup. In the EG, tooth brushing was initially taught and in the IA monitored and re-educated once a week by an occupational therapist. In contrast, the CG-IA group received a weekly placebo activity such as manicure by the same person. RESULTS: From the individual movements taught and monitored by the occupational therapist, opening a tube of toothpaste (n.s.) and denture brushing (p < 0.05) were performed more independently after 3 months. Both the occupational therapy and the placebo activity led to a significant improvement in oral (p < 0.01 and 0.05) and in denture hygiene (p < 0.001 and 0.05). From all participants, the EG-IA subgroup presented the most significant amelioration in plaque (p < 0.01) and denture hygiene scores (p < 0.001). This group consisted mostly of subjects with an impaired cognitive state. CONCLUSIONS: Despite the marked placebo effect, the results indicate that occupational therapy is particularly useful to improve the oral and denture hygiene in dependent and cognitively impaired LTC residents and may promote their autonomy in the execution of activities of daily life such as denture brushing. PMID- 15747896 TI - Age and perception of dental appearance and tooth colour. AB - OBJECTIVES: To determine the prevalence of satisfaction with dental appearance and own tooth colour in the UK and investigate the impact of ageing on these perceptions. METHODS: Data about ageing and satisfaction with dental appearance and tooth colour was gathered by means of a questionnaire completed at face-to face interview. The questionnaire was first tested and then used by the Office of National Statistics (ONS) in a stratified random sample of adults drawn from the UK. A total of 3384 adults were interviewed. RESULTS: Three quarters of the population were satisfied with their own dental appearance and two-thirds were satisfied with their tooth colour. Subjects over the age of 55 were significantly more likely to be satisfied with their dental appearance and tooth colour (p < 0.05) compared with younger age groups. CONCLUSIONS: Ageing is not necessarily associated with negative self-perception of dental appearance or tooth colour. Greater dissatisfaction with dental appearance or colour in younger ages may suggest that perceived appearance is linked to cognitive factors other than social or cultural ones. PMID- 15747897 TI - Adjunctive subantimicrobial dose doxycycline in the management of institutionalised geriatric patients with chronic periodontitis. AB - OBJECTIVE: To assess the efficacy of subantimicrobial dose doxycycline (SDD; 20 mg doxycycline twice daily) as an adjunct to scaling and root planing (SRP) in the treatment of moderate-severe chronic periodontitis (CP) in institutionalised elderly patients aged 65 years or older. BACKGROUND: Previous studies have shown that SDD is of clinical benefit in the treatment of CP. However, the benefits of SDD in geriatric populations (65+ years) have not been determined. MATERIAL AND METHODS: A 9-month, double-blind, randomised, placebo-controlled pilot study was conducted. 24 institutionalised geriatric patients (65 years or older) with evidence of CP manifested by baseline clinical attachment levels (CAL) 5-9 mm, probing depths (PD) 4-9 mm and bleeding on probing (BOP) were recruited. At baseline, patients were treated by a standardised episode of SRP, and randomised to receive either adjunctive SDD or placebo. Full mouth PD and CAL were measured using the manual UNC-15 periodontal probe at 3, 6, and 9 months post-baseline to assess the response to treatment. Periodontal sites were stratified by baseline PD value: sites with PD 4-5 mm were considered moderately diseased and sites with PD > or = 6 mm severely diseased. RESULTS: The SRP + placebo resulted in PD reductions similar to those reported previously in the literature. At all time points and in both moderate and deep sites, SRP + SDD resulted in significantly greater PD reductions relative to baseline than SRP + placebo. At month 9, in moderate sites, mean PD reductions of 1.57 +/- 0.11 mm were reported in the adjunctive SDD group, compared with 0.63 +/- 0.11 mm in the adjunctive placebo group (p < 0.001). In deep sites at month 9, mean PD reductions of 3.22 +/- 0.29 mm were reported in the adjunctive SDD group, compared with 0.98 +/- 0.31 mm in the adjunctive placebo group (p < 0.05). Similar improvements were observed for CAL in the SDD group compared with the placebo group. Significantly lower BOP scores were also recorded at month 9 in the SDD group (7.5%) compared with the placebo group (71.2%) (p < 0.01). CONCLUSION: SDD used as an adjunct to SRP provides significant benefit for elderly patients with CP compared with SRP alone. PMID- 15747898 TI - Number of teeth in relation to oral health behaviour in dentate elderly patients in Lithuania. AB - AIM: To describe oral self-care practices and the use of dental services among dentate elderly patients attending public dental service in Kedainiai, Lithuania, and relate these parameters to the number of remaining teeth. DESIGN: Cross sectional questionnaire study. SETTING: Two public dental offices in Kedainiai, Lithuania. PARTICIPANTS: Dentate patients aged 60+. METHODS: A self-administered questionnaire covering oral hygiene practices, use of sugar, utilisation of dental services, and number of teeth. Age, gender, and education served as background factors. RESULTS: In all, 174 dentate elderly people responded, their mean age being 69.2 years (CI 95% 68.2-70.2) and mean number of teeth reported 16.2 (CI 95% 15.4-17.1). Of these respondents, 30% reported that they brushed their teeth twice daily and 57% that they always used fluoridated tooth paste. Having 21+ teeth was strongly associated (p < 0.001) with these habits, with brushing in the evening, and with enrolment in higher education, but not with gender. After controlling for various self-care and background factors, frequent tooth brushing remained as the only significant factor in the logistic regression model (OR 2.0, CI 95% 1.2-3.3, p = 0.01) to explain subjects' retention of 21+ teeth. CONCLUSIONS: Elderly Lithuanians' oral self-care is far from meeting recommendations. To preserve their natural teeth, dentate elderly people should improve their oral self-care practices, in particular as regards tooth brushing. To reach that goal, all efforts by the community and dental profession are welcomed. PMID- 15747899 TI - Healthcare compunetics. AB - Changes in life expectancy, healthy life expectancy and health seeking behaviour are having an impact on the demand for care. Such changes could occur across the whole population, or for specific groups. Changes for specific groups will be particularly affected by policy initiatives, while both these and wider changes will be affected by people's levels of engagement with their health and the health service itself. Levels of education, income and media coverage of health issues are also important. These factors could also encourage an increase in people caring for themselves and their families or community. People are now expecting a patient-centred service with safe high quality treatment, comfortable accommodation services, fast access and an integrated joined-up system. The uptake of integrated Information and Communication technologies (ICT) will be crucial. Healthcare Compunetics, the combination of computing and networking customised for medical and care, will provide the common policy and framework for combined multi-disciplinary research, development, implementation and usage. PMID- 15747900 TI - The changes caused by modification of biomechanical properties of a bone. AB - The paper presents the distribution of the parameters of mechanical stiffness in a sample of animal bone. The parameters are measured at different distances from the cortical bone. As it is widely known, bone properties vary significantly depending on the length of the bone. Additionally, the properties of trabecular bone also depend on its distance from the cortical bone. The study presents the results for both samples of normal bone tissue and samples of bone tissue with chemically modified biomechanical properties. For each sample a matrix of measurement points was defined. For each measurement point, a micro hardness test was performed in order to define hardness of the micro volume and to measure the Young's modulus. This test was carried out using Micro-Combi-Tester equipment. Micro hardness was measured with Vickers diamond, and the analysis of deformation properties was realized by the Olivier and Pharr method. The application of Oliver and Pharr method enabled the calculation of Young's modulus directly from the unloading curve. Additionally, mineral density for all samples was measured using Dual Energy X-ray Absorptiometry method. This study allowed for comparison of bone mineral density parameter with Young's modulus. The aim of this comparison was to demonstrate changes caused by modification of biomechanical properties of bone. PMID- 15747901 TI - Web-based asthma collaboration management and public awareness. AB - Recent studies have shown that long-term monitoring of asthma severity can reduce asthma exacerbations, optimise drug therapy and decrease the cost of asthma management. The management of a chronic patient is a collective and cooperative enterprise that may exploit Information Technologies (IT) to improve the overall quality of care. The aim of this paper is to present a web based asthma tool that significantly enhances public information and awareness to support illness prevention, patients independent living through user profiling and personalisation and collaborative work between health professionals, therapists, caregivers and patients through Tele-Care and Tele-Consultation. The system has been tested through a preliminary survey that took place in UK and Greece. PMID- 15747902 TI - Extended possibilities of pharmaceuticals delivery to patients using dematerialized prescriptions. AB - This paper considers the requirement for e-commerce enabled delivery of pharmaceutical prescriptions. First, currently available solutions are critically reviewed and an ideal solution is specified and then the concept of the proposed solution--Dematerialized Prescription (DP), is outlined. Next, the information flows required to enable such a service are considered. The paper then considers a number of possible services which could be made available with DP to deliver medicals to patients. Finally, a proposed solution, which enables physician to fill dematerialized prescriptions online using inexpensive mobile Internet devices is presented in detail, the advantages of such a model are summarized and future research directions are suggested. PMID- 15747903 TI - Vibroacoustic sound therapy: case studies with children with profound and multiple learning difficulties and the elderly in long-term residential care. AB - This paper describes the development of Vibroacoustic Sound Therapy, an approach which is being developed for use in special schools for children with profound and multiple learning difficulties, and in long-term care homes for the elderly and the elderly mentally infirm. Using non-invasive techniques and music/sound technology, children and the elderly are being empowered and enabled to (re)discover self expression and communication skills. Two case studies drawn from the world of the young disabled, and from the elderly, illustrate the potential for improving quality of life and well-being in these institutional settings. PMID- 15747904 TI - An implementation of the trigram phrase matching method for text similarity problems. AB - The representation of texts by term vectors with element values calculated by a TFIDF method yields to significant results in text similarity problems, such as finding related documents in bibliographic or full-text databases and identifying MeSH concepts from medical texts by lexical approach and also harmonizing journal citation in ISI/SciELO references and normalizing author's affiliation in MEDLINE. Our work considered "trigrams" as the terms (elements) of a term vector representing a text, according to the Trigram Phrase Matching published by the NLM's Indexing Initiative and its logarithmic Term Frequency-Inverse Document Frequency method for term weighting. Trigrams are overlapping 3-char strings from a text, extracted by a couple of rules, and a trigram matching method may improve the probability of identifying synonym phrases or similar texts. The matching process was implemented as a simple algorithm, and requires a certain amount of computer resources. An efficiency-focused C-programming was adopted. In addition, some heuristic rules improved the efficiency of the method and made it feasible a regular "find your scientific production in SciELO collection" information service. We describe an implementation of the Trigram Matching method, the software tool we developed and a set of experimental parameters for the above results. PMID- 15747906 TI - Mobile guidelines for pharmacologic treatment of hypertension. AB - Mobile health technologies provide case studies with potentially rather huge impacts. Examples are provided e.g. by guidelines involving pharmacologic treatment. Knowledge and reasoning need to interact with information management, and often involves utility of various devices. Well organised databases for pharmacological information are also necessary for successful engineering of mobile extensions in these case studies. PMID- 15747905 TI - An electronic patient record implementation using clinical document architecture. AB - Electronic patient records (EPRs) provide the means for integrated access to patient information that may be scattered across dispersed healthcare organizations that, in general, use heterogeneous systems in order to support their internal functions. XML language and Clinical Document Architecture (CDA) provides a mechanism for defining, structuring, manipulating and visualizing patient medical data using the same semantics through web. In this paper, a prototype implementation of a web-based electronic patient record (EPR) system using XML for data format and CDA for defining and structuring patient clinical documents is presented. PMID- 15747907 TI - Artificial neural networks as an engine of Internet based hypertension prediction tool. AB - Hypertension is the most common cause of death. Therefore it is recognized as a major civilization disease next to diabetes, hyperuricemia, asthma etc. The objective was to use artificial neural networks (ANNs) to handle demographic data and to produce system of hypertension risk prediction. Database used in the development of hypertension risk model was obtained from CDC (BRFSS--Behavioral Risk Factor Surveillance System). Screening for optimal ANN architecture was performed among various backpropagation and fuzzy neural networks with use of 10 fold cross-validation scheme. Single ANNs as well as experts committees were tested. Best results were found to be around 75%--expressed as total classification rate. Java applet was designed to be the interface between ANN system and end user. Spreadsheet form was chosen to facilitate navigation and used by healthcare non-specialists. Free of charge Internet publication is expected soon at the address [url: see text]. PMID- 15747909 TI - Development of an expert system for differentiating tension type headaches from migraines. AB - We created a program, written in the CLIPS language for expert systems, to distinguish migraines from headaches. The system can be web-based, takes seconds to input data and quickly converges to the correct diagnosis. Over time such an application can result in enormous savings to doctors, patients, and health care institutions. PMID- 15747908 TI - Development of a clinical pathways analysis system with adaptive Bayesian nets and data mining techniques. AB - The use and development of software in the medical field offers tremendous opportunities for making health care delivery more efficient, more effective, and less error-prone. We discuss and explore the use of clinical pathways analysis with Adaptive Bayesian Networks and Data Mining Techniques to perform such analyses. The computation of "lift" (a measure of completed pathways improvement potential) leads us to optimism regarding the potential for this approach. PMID- 15747910 TI - Quality management issues for medical ICT. AB - Information and Communication Technology (ICT) assist healthcare professionals in the process of diagnosis, treatment, monitoring, medication prescription, referral, information retrieval and communication, documentation and transactions. This applies to intramural, transmural and extramural conditions. About 67% of patients involved in safety incidents believe that the incidents could have been prevented had an electronic health record been available. However, preliminary research also suggests that about 30% of patient safety incidents relate to software problems. ICT might improve quality of healthcare provided that ICT products, the ICT administration and use comply to essential requirements. This paper presents information about the state of the art of software quality, the measuring of software quality, quality management systems and a evaluation and certification method. PMID- 15747911 TI - MET system: a new approach to m-health in emergency triage. AB - The MET (Mobile Emergency Triage) system is an m-health application that supports emergency triage of various types of acute pain at the point of care. The system is designed for use in the Emergency Department (ED) of a hospital and to aid physicians in disposition decisions. Given patient's condition, MET recommends a triage by consulting decision rules stored in the system's knowledge base. The rules have been created using a data mining method (based on rough set methodology) applied to data collected during a retrospective chart study and verified by the clinicians. MET is designed following the extended client-server architecture, suited for weak-connectivity conditions, where stable connection between clients and a server cannot be provided. The MET server interacts with the hospital's patient information system in order to retrieve information about patients admitted to the ED. It also stores current patients' demographic and clinical data to be exchanged with mobile clients. The MET mobile client, running on a Personal Digital Assistant (PDA), is used for collecting clinical data and supporting triage decisions. The support function runs solely on the client side, thus it can be invoked anytime and anywhere, even if there is no communication link with the server (e.g., there is no wireless network available in the ED). Due to implementation on PDAs and working in weak-connectivity conditions, the MET system is very well suited for use in the ED and fits seamlessly into the regular clinical workflow without introducing any hindrances or disruptions that are often reported when using stationary (i.e., working on desktop computers) clinical systems. The system facilitates patient-centered service and timely, high quality patient management. It provides recommendations using a limited amount of clinical data, normally available at the point of care. Furthermore, it provides a possibility for the structured evaluation of this data by an attending physician. PMID- 15747912 TI - Robotic technology applied to meet the requirements of tools to support microsurgery and cellular surgery. AB - Microsurgery and processes involving cell manipulation or cell surgery are clinical practices where the operator works at or beyond the threshold of human dexterity. Current tools available are conventional in their design, and this limits consistency and the level of reliability and achievement. Surgical robotic devices have been explored to improve precision in minimal access surgical procedures to augment control of tool points in tissues, and have enabled feedback of sensory data from which the operator is able to deduce information on the tool at the working site. In this paper, relevant technologies are described that can be harnessed to improve perception of tool point interaction with tissues at the working site and to improve tool control at the small scale required in clinical practice for microsurgery and for cell surgery. PMID- 15747913 TI - Emotional Prosody Measurement (EPM): a voice-based evaluation method for psychological therapy effectiveness. AB - The voice embodies three sources of information: speech, the identity, and the emotional state of the speaker (i.e., emotional prosody). The latter feature is resembled by the variability of the F0 (also named fundamental frequency of pitch) (SD F0). To extract this feature, Emotional Prosody Measurement (EPM) was developed, which consists of 1) speech recording, 2) removal of speckle noise, 3) a Fourier Transform to extract the F0-signal, and 4) the determination of SD F0. After a pilot study in which six participants mimicked emotions by their voice, the core experiment was conducted to see whether EPM is successful. Twenty-five patients suffering from a panic disorder with agoraphobia participated. Two methods (story-telling and reliving) were used to trigger anxiety and were compared with comparable but more relaxed conditions. This resulted in a unique database of speech samples that was used to compare the EPM with the Subjective Unit of Distress to validate it as measure for anxiety/stress. The experimental manipulation of anxiety proved to be successful and EPM proved to be a successful evaluation method for psychological therapy effectiveness. PMID- 15747914 TI - Errors in medical practice: identification, classification and steps towards reduction. AB - We present a new taxonomy of medical errors, with emphasis on human errors. We illustrate errors due to medication, errors due to diagnosis, errors due to hospital treatment procedures, and errors related to clerical procedures. We also discuss a database of 143 papers on medical errors which we have developed. PMID- 15747915 TI - Using the sense of smell in telemedical environments. AB - It is widely recognized in the medical community that the sense of smell plays a very important role in the diagnosis of various associated medical disorders. With the advent of computerized scent detection and scent production devices, smell as a medical diagnostic and educational tool could be carried over into the world of telemedicine, tele-surgery, and distance education. PMID- 15747916 TI - Power to the patient, using DI@L-log. AB - Chronic care patients are demanding to be more actively involved in the care of their condition. Self-monitoring of blood glucose and blood pressure is strongly advocated for people with type 2 diabetes. We are developing a DI@L-log system that replaces the traditional paper logbook used by diabetes patients, enabling them to send their data to the point of care on a weekly basis using spoken dialogue technologies over the telephone. The motivation for our system is to enhance care by providing patients with a voice and therefore empowering individuals to become active self-managers of diabetes. PMID- 15747917 TI - Interactive telemedicine as a tool to avoid a digital divide in the world. AB - An efficient way of closing the gap in medical care between the EU and its Mediterranean partners is the use of satellite-based networks for telemedical applications. Using standard hardware components and specially designed communication software (WinVicos) various telemedical applications (teleconsultation, telementoring, teleteaching, etc.) have been realized. Two telemedical networks which have been designed and developed recently, MEDASHIP (Medical Assistance for Ships) and EMISPHER (Euro-Mediterranean Internet Satellite Platform for Health, medical Education and Research), are presented. PMID- 15747918 TI - Bioavailability and bioequivalence trials: statistics & pharmacokinetic principles. AB - This document has been prepared as a set of workshop material, which will be presented in the International Congress on Medical and Care Compunetics in June 2004 in Den Hague, The Netherlands. The workshop is divided into two parts and deals with the fundamentals of the bioavailability and bioequivalence studies. First part of the workshop deals with the pharmacokinetic principles, and the second part discusses the statistical approaches and the data analysis using the Statistical Analysis Software (SAS). PMID- 15747919 TI - The impact of structuring multidisciplinary team conferences mediated by ICT in the treatment of patients with rheumatic diseases. AB - Communication processes are pervasive in the daily practice of health professionals. Reorganizing these daily practices by introducing ICT, inevitably effects one or more communication processes. Understanding exactly what these effects are, is a major problem in designing and implementing ICT-applications. In this paper we present an analysis of these effects, based on a theory of communication processes. The concept of 'decoupling' is pivotal in our analysis. Based on the identified effects, and some preconditions that have to be met in order for these effects to take place, we derive a number of guidelines for reorganizing communication processes by means of applying ICT. The application of these quidelines will be demonstrated and discussed in the context of the reorganization of a team conference at the Rehabilitation Clinic of the Rheumatology Department of the Leiden University Medical Center (RCRD/LUMC), a multidisciplinary team care setting. PMID- 15747920 TI - Enhanced communication between care professionals and patients with hemophilia by using a web enabled electronic logbook. AB - Hemophilia is an X-linked clotting disorder. Treatment consists of intravenous substitution of the missing clotting factor. Although very effective, clotting factor products are very expensive. Most of the clotting factors are administered at home by the patients themselves. Documentation of the amount used and reasons for administrating of these scarce and expensive clotting factors in a paper logbook is a problem. The use of an electronic logbook results in improved compliance in documentation of clotting factor use and therefore in improved quality of treatment of patients with hemophilia. PMID- 15747921 TI - Tele-homecare for chronically ill persons: pilot trials, medical outcomes and future perspectives. AB - Socio-economic trends including the ageing of populations in the developed world have let to markedly increased interest in improving society's ability to deliver effective care to older and chronically ill patients at home. The paper reports progress from recent European research on three key fronts faced in the diffusion of tele-homecare: Pilot applications to explore the features of as yet untried care domains, full clinical trials to present hard evidence of desired medical outcomes, and R&D to expand the capability of tele-homecare systems to meet the needs of a yet wider variety of chronic illnesses and patient groups. PMID- 15747922 TI - Remote training and monitoring. PMID- 15747923 TI - ICT applications for tele-consultation to increase efficiency and effectiveness of care. PMID- 15747924 TI - Information technology and behavioral medicine: impact on autism treatment & research. AB - Families, clinicians and researchers involved with varying neurological disorders face amazing challenges to understand, treat, and assist the people they are serving. Autism brings unique challenges and serves as an important model for the application of important concepts in information technology and telemedicine. The rising incidence of autism with limited professional resources has led to more consideration for using information technology and related specialties to link families and professionals, and to implement strategies which implement information technology to improve the outcomes for individuals with autism and their families. These are reviewed in context of the unique health, education, and the research issues facing those dealing with autism. PMID- 15747925 TI - The challenges of imaging based computational fluid dynamics. AB - Image based Computational Fluid Dynamics (CFD) simulation of the cardiovascular system is increasingly becoming important and its application in everyday medical practice can already be envisaged. The goal of this workshop is to address all the factors involved in the development of a computational framework/software for modelling and analyses of the cardiovascular system and provide examples. The development of such framework, requires integration, management and interpretation of data from several technology areas such as a) feature detection and extraction of arterial geometry from imaging data, b) adaptive grid generation techniques for 3-D asymmetric geometries c) hemodynamic modelling, disparate length-scale model, and fluid-tissue interaction with high-performance computing, d) CFD data validation, e) feature extraction/detection and visualisation algorithms, f) graphical user interface to allow remote visualisation of post processed data. These computational tools are employed to study flow in specific problem sites in the vascular tree such as the carotid, femoral, coronary and abdominal arteries. Such studies provide understanding of the factors involved in the initialisation and evolution of arterial disease due to altered flow conditions (as a result of plaque formation) such as flow separation and reversal, and low and oscillatory wall shear stress. It is also used to study the effect of various clinical procedures such as the implantation of stents, vascular grafts, vascular prostheses and artificial valve implants on local and global hemodynamics. This workshop will address a new emerging paradigm in clinical practice known as predictive medicine for effective surgical planning and post surgical rehabilitation. The workshop will also address the difficulties in the implementation of some of the technology areas in this application with examples of carotid, femoral, and abdominal artery simulations. PMID- 15747926 TI - Flow patterns and wall shear stresses in patient-specific models of the abdominal aortic aneurysm. AB - Abdominal Aortic Aneurysms (AAAs) are a major cause of death in Western societies. However, the factors responsible for the growth and ultimate rupture of AAA are not fully understood. Using computation fluid dynamics combined with CT images, the flow patterns and wall shear stress (WSS) in the AAA of a patient were studied. AAA geometry was obtained from images acquired during a routine CT scan, and a realistic abdominal aortic flow waveform was used as an upstream boundary condition. The computational results showed the presence of multiple flow recirculation zones and overall low WSS in the bulge of the aneurysm. This may provide some insight into the growth of thrombus in AAA. PMID- 15747927 TI - Vascular specialists training, the practice and the future. PMID- 15747928 TI - Quantitative analysis of vascular images, in particular of abdominal aorta aneurysms from 3D CTA data sets. AB - This article presents a combination of well known image processing techniques to automatically segment CTA images of the Abdominal Aortic Aneurysm. Current results are that about 80% of the contours need no manual corrections. The remaining 20% fail due to calcified plaque close to the lumen border. After correction a 3D surface model is created from the 2D contours which is used as input for flow simulations and for parameter extraction of the AAA by clinicians for selecting the proper size and shape endograft, and to plan the placement procedure of this endograft in the patient. PMID- 15747929 TI - The EAG tool: a decision support system for selection of abdominal aorta aneurysm endografts. AB - This article describes a decision support system (the EAG tool = Effective AAA Graftmanship) that assists the vascular surgeon in deciding whether a patient is fit for an endograft procedure and that assists the vascular surgeon in selecting the proper graft and in planning the endograft procedure. The EAG tool is the first step in the development of a learning environment for AAA procedures. As such the EAG tool is part of the VREST development project aiming to create a complete set of Virtual Reality Educational Surgical Tools. The EAG tool has been validated by using the expert knowledge of five experienced AAA physicians: three vascular surgeons and two interventional radiologists. In over 3,000 assessments, the EAG tool showed a false-positive rate of 0.2%, or in other words: in only 1 out of 500 cases the EAG tool indicated the possible use of an endograft, whereas the combined group of experts believed that it was not possibe. The EAG tool proved to be more prudent than the group of experts in having a false-negative rate of 3.7%. The EAG tool thus proved to be useful in daily clinical practice. Further developments are ongoing to use the EAG tool in a learning environment, specifically for unexperienced physicians. PMID- 15747930 TI - Stereopsis in medical virtual-learning-environments. AB - In surgical training a lot of effort is directed at developing virtual learning environments (VLE's). These environments usually include one or more of the following features: haptic feedback, the sending of a slightly different computer generated image to both eyes (affording an user's experience of stereoptical depth), and a transparant interface that allows 'natural' interaction. Now that many of these VLE's are approaching a state of maturity, time has come to start evaluating these environments, as basic questions regarding the effectiveness of the features mentioned above remain unanswered. The purpose of this article is to present an experiment aimed at evaluating the effects of stereopsis during a learning phase on visuo-spatial reasoning in two subsequent tasks that are related to the medical practice of diagnosis. PMID- 15747931 TI - Good healthcare innovation practice (GHIP)--a network-driven, knowledge-conscious approach for innovation in the (Dutch) healthcare sector. AB - In this article we describe why and how a new approach has been developed for innovation projects in the Dutch healthcare sector. There are three key aspects in the approach: Innovation should be seen as a (fair) process, the process is owned by all stakeholders (which calls for a Network approach) and, finally, the process should be knowledge-enabled, i.e. in each process step it should be clear what knowledge is needed, what knowledge becomes available and how the flow of knowledge should be managed. PMID- 15747933 TI - Cognition, interaction and ageing: an Internet workshops exploratory study. AB - Gerontology is known more and more as an interdisciplinary and functional knowledge. Geriatrics as one of its branches intends to make possible longevity with health. World population ageing occurs along with important economical and social inequalities for elder people, which are likely to be more affected by deficiencies (physical and/or cognitive) than younger groups. With the purpose of minimizing these deficiencies, Internet Workshops were accomplished, with a retired group of senior persons. This research was developed to bring together principles of Human-Computer Interaction, informatics, accessibility and gerontology in order to promote Digital Inclusion to this growing population and a methodology to virtual cognitive rehabilitation. PMID- 15747932 TI - Early detection of skin cancer (EDISCIM) through the use of non-invasive confocal imaging. AB - In the past 15 years the number of malignant melanomas and non-melanoma skin cancer, (i.e. squamous cell carcinoma, basal cell carcinoma), have increased dramatically throughout the whole world, in particular among people with white skin. For example, in Germany the number of new cases of melanoma is about 10 to 12 per 100,000 people, (for basal cell carcinoma as a non-melanoma skin cancer 140 per 100,000), the absolute number of annual new cases of melanoma is an estimated 9,000 to 10,000, and the increasing rate is 5 to 10%. However, there are more than 40 differential diagnoses for skin cancer which makes it difficult even for expert dermatologists to give correct diagnosis, (estimated 75% correct diagnoses). This figure is even worse for doctors from other specialities. There is only a chance of high cure rates when skin tumours are detected at an early stage. The prognosis of skin cancer is dependent very much on the thickness, (as a sign of invasiveness), of the tumour and the number of mitotic figures in the tumour, (as a sign of proliferation activity). However, the final diagnosis of skin cancer is usually done by biopsy, the sample being investigated by a specialist. This is an invasive method, which is painful for the patient and might require several cuts and samples taken before being absolutely certain. Due to the potential risk of dissemination of tumour cells, taking an incisional biopsy is obsolete in malignant melanoma. Therefore the correct diagnosis of malignant melanoma can be made only after a total removal of the lesion and consecutive histopathological examination. The EDISCIM project intends to develop a system for the early diagnosis of skin cancer which uses confocal imaging for the non-invasive diagnosis of the upper layers of the skin that aids the physician in the analysis of the images and therefore with the diagnosis, allows for potential remote diagnosis by specialist dermatologists via tele-diagnosis if need be. The objectives of the system are Captures microscopic images of the skin by confocal imaging, Processes and records these images in real time, Compares these images against a knowledge base of known skin symptoms, Displays the results in a suitable interface to the physician performing the diagnosis, And supports the online telematic support by specialists. PMID- 15747934 TI - A service for remote screening of physiological data and follow up of patients using a home gateway and domestic wireless technologies. AB - The availability and the reduced costs of more sophisticated electronic devices makes possible to develop and use sophisticated systems also for domestic use. This opportunity beside new emerging market needs is bringing to new life the Home-Care scenario. This paper will present a Home Care system developed in the context of the FUTUREHOME IST project. Its key features are interacting with external health-care service providers, such as the Hospitals, Private Clinics or Emergency Departments, from our own houses by means of innovative solutions. It enables the telemonitoring of vital parameters of the home/family members. Using wireless connections, data collected from different devices (weight scale, ox meter, glucose meter, etc...), are stored into the Home Network Service Point (HNSP), the FUTUREHOME network's core, from where they are used for further consultations (e.g. patients follow-up). PMID- 15747935 TI - Mobile patient monitoring: the MobiHealth system. AB - The forthcoming wide availability of high bandwidth public wireless networks will give rise to new mobile health care services. Towards this direction the MobiHealth project has developed and trialed a highly customisable vital signals' monitoring system based on a Body Area Network (BAN) and an m-health service platform utilizing next generation public wireless networks. The developed system allows the incorporation of diverse medical sensors via wireless connections, and the live transmission of the measured vital signals over public wireless networks to healthcare providers. Nine trials with different health care cases and patient groups in four different European countries have been conducted to test and verify the system, the service and the network infrastructure for its suitability and the restrictions it imposes to mobile health care applications. PMID- 15747936 TI - JUST in time health emergency interventions: an innovative approach to training the citizen for emergency situations using virtual reality techniques and advanced IT tools (the Web-CD). AB - This paper reports the results of the first of the two systems developed by JUST, a collaborative project supported by the European Union under the Information Society Technologies (IST) Programme. The most innovative content of the project has been the design and development of a complementary training course for non professional health emergency operators, which supports the traditional learning phase, and which purports to improve the retention capability of the trainees. This was achieved with the use of advanced information technology techniques, which provide adequate support and can help to overcome the present weaknesses of the existing training mechanisms. PMID- 15747937 TI - JUST in time health emergency interventions: an innovative approach to training the citizen for emergency situations using virtual reality techniques and advanced IT tools (the VR Tool). AB - This paper reports the results of the second of the two systems developed by JUST, a collaborative project supported by the European Union under the Information Society Technologies (IST) Programme. The most innovative content of the project has been the design and development of a complementary training course for non-professional health emergency operators, which supports the traditional learning phase, and which purports to improve the retention capability of the trainees. This was achieved with the use of advanced information technology techniques, which provide adequate support and can help to overcome the present weaknesses of the existing training mechanisms. PMID- 15747938 TI - Building knowledge driven and dynamically networked communities within European healthcare systems. AB - COCOON is an innovative project developed in the e-health area of the Sixth Framework Programme co-financed by European Commission. It is an Integrated Project aimed at supporting health care professional in reducing risk management in their daily practices by building knowledge driven and dynamically adaptive networked communities within European healthcare systems. PMID- 15747939 TI - Team work based care in speech and language therapy through web-based tools and methods. AB - Speech and language therapists (SLTs) are faced daily with a diversity of speech and language disabilities, which are associated with a variety of conditions ranging from client groups with overall cognitive deficits to those with more specific difficulties. It is desirable that those working with such a range of problems and with such a demanding workload, plan care efficiently. Therefore, the introduction of methodologies, reference models of work and tools, which significantly improve the effectiveness of therapy, are particularly welcome. This paper describes the first web-based tool for diagnosis, treatment and e Learning in the field of Language and Speech therapy. The system allows SLTs to find the optimum treatment for each patient, it also allows any non-specialist user--SLT, patient or helper (relative etc.)--to explore their creativity, by designing their own communication aid in an interactive manner, with the use of editors such as: configuration and vocabulary. The system has been tested and piloted by potential users in Greece and the UK. PMID- 15747940 TI - Cost-effective health services for interactive continuous monitoring of vital signs parameters--the e-Vital concept. AB - The objective of the e-Vital project is the validation of the market concerning the provision of a novel remote telemedicine service aimed at large sensitive parts of the European population, the "at-risk" citizens, who are usually patients with a stable medical condition that allow a near normal life but may suddenly deteriorate and put life at risk. This service will increase their quality of life and their feeling of safety concerning their health. The e-Vital project focuses on the implementation and exploitation of a modular and ambulatory secure telemedicine platform, which is using easily wearable vital signs monitoring devices, causing minimal discomfort to patients, and which transfer in real time and on-line critical vital parameters to doctors and/or medical experts/consultants, regardless of their location, while getting feedback to increase their feeling of comfort or in case of alarm. The interactive continuous monitoring promises cost effective health services, more active involvement of patients in their own care, and a new sense of realism in making a diagnosis. PMID- 15747941 TI - Ambulatory monitoring for chronic cardiac and pulmonary patients. AB - In the Greek pilot of the e-Vital project, remote telemedicine services are provided to chronic cardiac and pulmonary patients who are not confined to a hospital (i.e. receiving home/ambulatory health care) with the aim of exploring the dynamics of interactive continuous chronic patient monitoring, particularly focusing on the impact on patient's quality of life, the patient's active involvement in their own care and according impact on the overall quality of healthcare provision, as well as the benefits for healthcare providers (time management, patient management, savings, etc). The current trial in progress validates the business potential of remote monitoring services and demonstrates the technical environment that enables patient-doctor interaction regardless of location and the according communication modes and protocols. The domain for applying the envisaged service is the private healthcare sector and the users groups include individual chronic patients with cardiac and pulmonary diseases. So far, from the patient's point of view, the service mainly appeals to patients with arrhythmias due to lightweight technology and easy processes involved in the transmission of 1-lead ECG. Asthma and COPD patients are also enthusiastic, as they can receive doctor advice in real time. From the health professional's point of view the service has real added value in the fields of diagnosis, prevention, monitoring and follow up. In the case of asthma and COPD the value of the service in diagnosis is even more apparent as these patients may be asymptomatic, when they visit their doctor and have exacerbations when they are at home. e-Vital in this case ensures that the healthcare professional will acquire the complete image of the patient condition. Finally, the experiences gained so far indicate that e-Vital could be particularly efficient in isolated areas, with shortage of experienced scientific personnel. PMID- 15747942 TI - A new model for home care for COPD. AB - A new model for home care of COPD patients is investigated, as a part of a coordinated provision model across levels of care. In the Spanish pilot of the e Vital project, relevant vital signs for COPD are closely monitored and used for early detection of deterioration in the state of the patient and all prompt treatment. This can also reduce the need for in-person check-ups and re-admission to hospital. Later analysis aims to determine whether the system has significant economic impact, and whether it will provide more convenience for patients and healthcare professionals. The present trial focuses on the impact of community based nurses visiting each patient in the home as a part of this shared care. The visits are part of the regular follow-up for patients with chronic conditions who belong to any of the home care programs currently available in our institution. In each one of these visits, the nurse performs a number of tasks in order to assess patient status. Vital signs are collected and may be transmitted to the data monitor centre automatically using 2.5-3G technology. Sensors communicate through a Body Area Network (BAN) with a PDA configured to display the data and transfer it to the server using GPRS. Patients can be trained to use the equipment on their own. Results so far are encouraging. In the previous phase, a similar set-up without monitoring facilities at the patient's home showed improvements in several clinical indicators (ER visits, SGRQ, Quality of life, LOS and costs) for a home hospitalisation program and in a prevention of exacerbation program. The current set-up aims at increasing such benefits and further extending the target population. PMID- 15747943 TI - A new monitoring service for long term residential care. AB - eVital, a European funded project, aims to investigate the issues in the provision of vital signs monitoring services in the community. For the UK pilot, services to monitor the vital signs of patients in residential care homes by remote health care workers have been established. In particular, the aim was to allow the resident's own GP to observe the ECG, blood pressure, SpO2, temperature and respiration whilst still in the health centre and be able to advise on the most appropriate action should a crisis occur. As outcomes, this pilot project was designed to determine feasibility, channels of communication between key players and assess the technology. Trials have been under way for four months and preliminary results are promising. Residential home staff have commented on the extra level security they feel in being able to "summon" medical support for residents for whom they may have concern. Patient's relatives have also commented on the reassurance they experience from knowing that the resident is being monitored. PMID- 15747944 TI - A reference architecture for telemonitoring. AB - The Telecare Interactive Continuous Monitoring System exploits GPRS to provide an ambulatory device that monitors selected vital signs on a continuous basis. Alarms are sent when parameters fall outside preset limits, and accompanying physiological data may also be transmitted. The always-connected property of GPRS allows continuous interactive control of the device and its sensors, permitting changes to monitoring parameters or even enabling continuous monitoring of a sensor in emergency. A new personal area network (PAN) has been developed to support short-range wireless connection to sensors worn on the body including ECG and finger worn SpO2. Most notable is use of ultra low radio frequency to reduce power to minimum. The system has been designed to use a hierarchical architecture for sensors and "derived" signals, such as HR from ECG, so that each can be independently controlled and managed. Sensors are treated as objects, and functions are defined to control aspects of behaviour. These are refined in order to define a generic set of abstract functions to handle the majority of functions, leaving a minimum of sensor specific commands. The intention is to define a reference architecture in order to research the functionality and system architecture of a telemonitoring system. The Telecare project is funded through a grant from the European Commission (IST programme). PMID- 15747945 TI - MDA-based EHR application security services. AB - Component-oriented, distributed, virtual EHR systems have to meet enhanced security and privacy requirements. In the context of advanced architectural paradigms such as component-orientation, model-driven, and knowledge-based, standardised security services needed have to be specified and implemented in an integrated way following the same paradigm. This concerns the deployment of formal models, meta-languages, reference models such as the ISO RM-ODP, and development as well as implementation tools. International projects' results presented proceed on that streamline. PMID- 15747946 TI - Implementing MDA-based distributed, interoperable, flexible, scalable, portable, and secure EHR systems. AB - Electronic Health Record (EHR) systems provide the kernel application of health information systems and health networks which should be independent of complexity, localisation constraints, platforms, protocols, etc. Based on shared care information systems' requirements for high level interoperability, a generic component architecture has been introduced. For implementing, running and maintaining acceptable and useable health information systems components, all views of the ISO Reference Model-Open Distributed Processing have to be considered. Following the Model Driven Architecture paradigm, a reference model as well as concept-representing domain models both independent of platforms must be specified, which are combined and harmonised as well as automatically transferred into platform-specific models using appropriate tools. PMID- 15747947 TI - A peculiar aspect of patients' safety: the discriminating power of identifiers for record linkage. AB - Today, one of the main stake of the interfaced health information systems or networks is to be able to gather the different parts of the medical record of a patient without any risk to mix them with those of one other patient. This objective could appear easy to reach but only in theory because in practice many name are misspelled or erroneous and a great attention has to be paid to define what is the best identifier to link medical record. As a linkage using less informative identifiers could lead to linkage errors, it is essential to quantify the information associated to each identifier. The aim of this study was to estimate the discriminating power of different identifiers susceptible to be used in a record linkage process. This work showed the interest of three identifiers when linking data concerning a same patient using an automatic procedure based on the method proposed by Jaro; the date of birth, the first and the last names seemed to be the more appropriate identifiers. Including a poorly discriminating identifier like gender did not improve the results. Moreover, adding a second christian name, often missing, increased linkage errors. On the contrary, it seemed that using a phonetic treatment adapted to the French language could improve the results of linkage in comparison to the Soundex. However, whatever, the method used it seems necessary to improve the quality of identifier collection as it could greatly influence linkage results. PMID- 15747948 TI - High level security policies for health care establishments. AB - Health Care Establishments (HCE) are today highly dependent upon Information and Communications Technologies (ICT). This increasing reliance upon ICT has stressed the need to foster security in Healthcare Information Systems (HIS). Security policies may have a significant contribution to this effort, but they could become the cause of portability and interoperability problems. Moreover, policies that fail to take into account all the aspects of HIS security, the legal and regulatory requirements, and the existence of several stakeholders may lead to ineffective or inefficient security measures. Policies of a special category, named Generic Security Policies (GSP), should be developed to provide policy level harmonisation and guidance to policy-makers within HCE. Six such policies are comparatively reviewed herein. PMID- 15747949 TI - High level security policies for health: from theory to practice. AB - The design and implementation of a security policy for a healthcare organisation is by no means trivial but it is, at least, feasible, taking into account the wide range of information security and privacy enhancing technologies that are currently available. Considering, however, a shared care environment with the participation of many independent healthcare organisations and the requirement for exchanging electronic healthcare records, the situation becomes much more complex since the implementation of global security policy may turn out to be an over ambitious task. This paper aims to highlight the main sources of complexity and to provide pointers for managing or/and resolving them. PMID- 15747950 TI - Access control management in practical settings. AB - Looking at the subject of access control from a practical side, focussing on the electronic health record, we discuss two special cases: the need of providing emergency access, and the possibilities of having a completely automated access control facility. We conclude that the first is essential, and suggest that the latter might not even be desirable. PMID- 15747951 TI - Policy management and access control in practice. AB - This paper reflects how the combined CEN and ISO standard Privilege Management and Access Control (PMAC) Part 1 to 3 is intended to work. It also gives experiences from a test implementation. PMID- 15747952 TI - Security infrastructure services for electronic archives and electronic health records. AB - Communication and co-operation in the domain of healthcare and welfare require a well-defined set of security services based on a Public Key Infrastructure and provided by a Trusted Third Party (TTP). These services describe both status and relation of communicating principals, corresponding keys and attributes, and the access rights to applications and data. Additional services are needed to provide trustworthy information about dynamic issues of communication and co-operation such as time and location of processes, workflow relations, and system behaviour. Legal, social, behavioural and ethical requirements demand securely stored patient information and well-established access tools and tokens. Electronic (and more specifically digital) signatures--as important means for securing the integrity of a message or file--along with certified time stamps or time signatures are especially important for purposes of data storage in electronic archives and electronic health records (EHR). While just mentioning technical storage problems (e.g. lifetime of the storage devices, interoperability of retrieval and presentation software), this paper identifies mechanisms of securing data items, files, messages, sets of archived items or documents, electronic archive structures, and life-long electronic health records. Other workshop contributions will demonstrate related aspects of policies, patient privacy, and privilege management. PMID- 15747953 TI - Secondary use of the EHR via pseudonymisation. AB - The Electronic Health Record (EHR) has many secondary uses, such as health economy and health care research, or disease specific clinical or epidemiological research. For these uses in general the patient identity is not needed, therefore the data must be anonymised or pseudonymised. Whereas for one-time use of the data this procedure is straightforward, long-term data accumulation or the necessity of re-identification require a more sophisticated approach. This paper describes possible model architectures, developed for medical research networks, but useful in other contexts as well. PMID- 15747954 TI - Use of the ISO/IEC 17799 framework in healthcare information security management. AB - Shared care implies sharing information. This requires a common concept of information security among healthcare providers and a system to maintain compliance to the security requirements within the healthcare community. This paper describes the use of the Code of Practice for Information Security Management ISO/IEC 17799 as a general framework for establishing a set of controls for information security in a particular organisation and as a framework for standards on information security in healthcare and their implementation. PMID- 15747955 TI - Security requirements in EHR systems and archives. AB - EHR system is a system for recording, retrieving, and manipulating information in electronic health care records. Archive is an organisation that intends to preserve health records for access and use for an identified group of consumers. There exist many combinations of EHR-systems and archives. EHR-system can be a single on-line system with integrated archiving functions or archive and EHR system are co-operative or federated systems. This paper describes both common security requirements for EHR-systems and archives and security requirement specific for archives. Requirements are derived from ethical and legal principles. From principles a set of security requirements are derived. Safeguards for implementing security are discussed. In practise EHR-system and archive share many security services. This document is proposing that inside a security domain both the archive and EHR-system have a common security policy. In addition to this the archiving organisation needs a documented policy for information preserving and a policy for access and distribution of information between other archives. PMID- 15747956 TI - Electronic health record on cards. AB - The paper is dedicated to the brief evaluation of smart card technology in terms of implementing EHR. Basic standards and recommendation are mentioned, so as currently running European activities. It has to be stressed, that despite the health care (and social insurance) is controlled throughout EU on national basis, strong international harmonisation and interoperability is being perceived. EFMI WG Cards contribution in this sense is drafted as closing remark to the paper. PMID- 15747957 TI - Non-telephone healthcare: the role of 4G and emerging mobile systems for future m health systems. AB - The next generation of "m-health technologies" is a new and evolving topic in the areas of telemedical and telecare systems. These technologies involve the exploitation of mobile telecommunication and multimedia technologies to provide better access to healthcare personnel on the move, by removing the key disadvantage of trailing wires in current systems. These technologies provide equal access to medical information and expert care by overcoming the boundaries of separation that exist today between different users of such medical information. A great benefit to all users will be a more efficient use of resources and far greater location independence. In this paper we will address some notes and future trends in these emerging areas and their applications for m health systems. Especially we will discuss the role of 4G and emerging mobile systems for future m-health systems. The new technologies can make the remote medical monitoring, consulting, and health care more flexible and convenient. But, there are challenges for successful wireless telemedicine, which are addressed in this paper. PMID- 15747958 TI - Telemedicine development: setting the record straight. AB - When a topic so synonymous with advanced technology as telemedicine needs a written history, perhaps a discipline has come of age. Telemedicine has had an oral history among visionaries and technophiles over the last thirty years or so. Tales of grand projects, technical failures and fabulous successes abound at telemedicine meetings. One is reminded of the film footage of early efforts at flight where extravagant engineering approaches failed time and time again until the elegant solution of the Wright brothers took to the air on the beach at Kitty Hawk in December 1903. It is hard to imagine the frustration and excitement of initial efforts at telemedicine or manned flight when current expectations are for information on demand and on time departures. However, from very early efforts in telemedicine the vision was consistent: overcome the huge liability of distance in patient care by creating an electronic continuum between the site of need and the site of expertise. The vision continues to strive toward technical transparency and clinical results, which assure a patient that no matter who, no matter where, no matter when medicine will care for you with consistent professionalism. Technology took us from the patient home to the technology-rich hospital. Perhaps technology will transfer the majority of patient management back to the comforts of home, convenience of workplace or the exigencies of travel and mobility. PMID- 15747959 TI - Instead of a prologue. Establishing telemedicine in the Balkans: from Berlin to Prishtina via Mars--a personal journey. PMID- 15747960 TI - Changing the medical world order with technological advances: the future has only begun. AB - It is a great honor to contribute in a small way to this book which in a way celebrates the continued evolution of telemedicine in Kosova. I was privileged to visit this brave land on several occasions in the last years. Severe circumstances were repeatedly resolved through great personal and professional sacrifice. Revolutionary solutions have been applied where evolutionary development was simply too slow in the realization of a new land in a complex world. In some ways telemedicine is emblematic of such struggle and success. We aspire to world health when world peace is elusive and hunger haunts us in the midst of plenty. Telecommunications have made it easy to report the failings of medicine, the injustice of health care and the unmet promise of political endeavor. It is the promise of telemedicine to use those same channels of information to empower, unify and advance the cause of health rather than only report the failings. Telemedicine entails the use of telecommunications and information technology to support the delivery of health care at a distance. There are critics who believe telemedicine is a waste of precious resources, which are needed urgently for higher health priorities. Telemedicine is dismissed as an expensive irrelevance, another distraction from the real needs of medicine in a chaotic world. That is patently ridiculous. Telemedicine is a part of the wider phenomenon of information and information is arguably the strongest change agent in play for medicine and other societal elements as well. A well-informed public armed with the tools for self-determination and the evidence for efficient action cannot be corrupted. Telemedicine is a part of the great change information brings to the world order, a drastic change toward a better world of health and justice. PMID- 15747961 TI - E-leadership in the digital age: the equilibrium, the codes and the digital gap. AB - Malaysia's experience in implementing the Integrated Telehealth Project has placed her way ahead in the arena of world Telehealth. Thus, she has become the focus point, reference point and benchmark for similar endeavors around the world. In fact, it would not be presumptuous to state that the Integrated Telehealth project is a trail-blazing pioneer with e-leadership experience and skills developed over the last few years. It is hoped that the Integrated Telehealth concept will find acceptance and credence globally. PMID- 15747962 TI - Challenges and barriers to development of telemedicine programs. AB - The collection of data is a vital step in assuring that a system works, a method is correct and the process is adequate. As data is collected databases are created. These databases could be nothing more than a box or it could be in a computer-based, indexed database. Over the past 200 years, technology has played an increasingly larger role in health care delivery. With the advent of telecommunications capabilities and information systems, medicine has embraced these two disciplines, resulting in a new tool - telemedicine. This new tool is undergoing rigorous peer review and scientific challenge. This has lead to a plethora of data, which indicates telemedicine is an excellent tool for medicine. PMID- 15747964 TI - Ten critical steps for a successful telemedicine program. AB - This chapter is based on a study of the characteristics that drive successful and unsuccessful telemedicine programs. In 2000 the International Society for Telemedicine (ISfT) asked AMD Telemedicine to present our observations of what characteristics appeared to be common to successful telemedicine and telehealth programs. To find out, AMD Telemedicine conducted a study of over 60 telemedicine programs in three countries. During this review, we identified ten basic points and documented successful and unsuccessful approaches to each. There are exceptions, but the approach to each of these issues appears to maximize the likelihood of success or failure. The findings were initially presented to the ISfT membership at their conference in Denmark. The study has been updated continuously as new information becomes available. The most current observations are as follows. PMID- 15747963 TI - Creating telehealth networks from existing infrastructures. AB - Implementing a telehealth program can be a financial challenge due to associated costs such as hardware, software, networking, administration and clinical expertise. To maximize potential and minimize costs, it makes sense to leverage existing telecommunications infrastructures and tailor the program based on what is available. PMID- 15747965 TI - Data mining challenges in the design of telemedicine platforms. AB - The evolution of telemedicine information systems involves the general processes of acquiring useful knowledge from medical data sets for diagnosis, intelligent efficient patient record transmission and autonomous adaptation of biomedical devices and their related software environments using quality of service attributes. Knowledge engineering concepts and methods allow application to the design of intelligent telemedicine platforms, satisfying the performance requirements and the quality assurance criteria of each specialized telemedicine application. PMID- 15747966 TI - Integrated telehealth: putting it all together and addressing the challenges. AB - The Integrated Telehealth Project of Malaysia is considered a principal enabler for the nation's Vision 2020 as well as the National Health Vision. Being in such an unenviable position, of being not only the pioneer for such an integrated project, but also with no benchmark to compare with, the project implementers have faced manifold challenges along the way. This chapter deals with some of the challenges and lessons learnt that have accumulated as the project progressed. PMID- 15747967 TI - Telemedicine models for primary care. AB - The provision of healthcare from the distance is extremely important in primary care for all nations in the world and especially as a humanitarian response for developing countries. International experience has demonstrated the clinical value and the technical feasibility of telemedicine-based patient care and service collaboration in primary care. Various models are proposed herein on the basis of the available technological driving forces, arrangements and current practices. PMID- 15747968 TI - Experiences and perspectives in using telematic prevention on sensitive health issues. AB - The new information and communication technologies, telematics - such as the Internet, telephone services and videoconferencing - are simultaneously both an instrument and a symbol - a sign of progress - but also a potential addiction problem. Sensitive topics - like substances or mental health - bring out all these characteristics of telematics. Therefore the computer world, substances and addictions are closely connected. PMID- 15747969 TI - Telemedicine in trauma and disasters--from war to earthquake: are we ready? AB - Every year many disasters cause thousands of injuries, deaths, refugees. Earthquakes and war often cause severe injuries (burns; amputations; Crush Syndrome; gunshots; landmines; nuclear, biological or chemical warfare / hazardous material; infectious diseases; pediatric specialties). Referring to big earthquakes in the last few years up to 20.000 thousand people were killed (India 2001). 310.000 deaths were caused by war in 2001. The Mass Casualty Incident is characterized by the disbalance between victims and the normal community emergency response. Because of this a lot of different institutions and organizations are involved in coping with the disaster. This produces an extensive demand of qualified Command, Control and Communication (C3). Furthermore a lot of data has to be collected during the treatment and the injuries need special medical treatment. The use of health telematics in disaster response helps to cope with the scenario. Modern technologies provide support for building up medical aid although the normal infrastructure is destroyed. To cope with disaster scenarios there are some telematic tools which can be used: computer based Command and Control System, telemedical support, and data resources-network/Medical Intelligence. The International Center for Telemedicine at the University of Regensburg Medical Center provides support for Health Care Professionals as a competence center for telemedicine. For the eastern part of Bavaria it develops a telemedical network with many components: The mobile emergency care system NOAH (Notfall-Organisations- und Arbeits-Hilfe) supports the Emergency Medical Service. Local Health Networks and the Clinical Network of Eastern Bavaria connect physicians and hospitals with the Regensburg Medical Center. With an online-education tool participants from all over the country can take part in trainings and courses. PMID- 15747971 TI - Home health and telemedicine: where are we today? AB - This chapter will review the current state-of-the-art of home health services in the telemedicine environment. Special attention will be paid to some of the studies that our telemedicine program has conducted in the past few years. The chapter will conclude with some recommendations and a look to the future. PMID- 15747972 TI - Education and distance learning: changing the trends. AB - Training and instruction are activities deeply ingrained in human relations and derive from the critical need for the young to learn survival skills. The responsibility in primitive society for such training almost certainly fell to parents who continued their pedagogical role after childhood issues to include hunting, gathering, fine motor activities and other life skills needed for personal or family survival. Such instruction only ended when the young were ready for independent life and- contribution to tribal well-being. Delegation of teaching to others was inevitable. Teaching has become a specialty and has at least one interesting story in ancient literature. Ulysses was certain to be away at the Trojan War and subsequent adventures for many years. He would not be able to provide his son, Telemachus, with the guidance and training to prepare him for adulthood. Therefore, he asked Mentor to act In Loco Parentis and instruct the young man toward competence and adult success. Teaching as a profession and discipline has been through many stages and many controversies. Socrates was a great teacher with a distinct technique for learning by questioning. His influence on his students was profound. Plato was such a good student he recorded all the master's works. Socrates has never been credited with even the briefest lecture note. As great as he was Socrates was forced to drink the bitter hemlock because his teaching was considered a corruption of youth rather than a proper preparation for effective adulthood. Dissonance between the expectations of learners, parents and teachers has a rich history. Certainly even now education is not something the professoriate may invent for the naive learner and then expect grateful acquiesce with faithful learning. Learning has -dimensions in human psychology and communication. The learners' autonomy, privacy and motivation cannot be denied. Learning is collaboration with teacher and the endpoint is the acquisition of new knowledge or skill. PMID- 15747970 TI - Role of telemedicine in neurosciences. AB - That distribution of specialists in neurological sciences worldwide is lopsided is an accepted fact. That, increasing the number of specialists, providing them with the requisite infrastructure and maintaining high standards to provide neurological and neurosurgical surgical care worldwide is just not possible, is also accepted. Patients from suburban areas are sent to the limited number of tertiary neurosciences referral centers, incurring expense and time and resulting in overwork at these centers. It is easier to set up an excellent telecommunication infrastructure, to increase the reach of the limited number of urban specialists, than to put them in suburban areas. PMID- 15747973 TI - From blackboard to whiteboard: the old style of classroom education is dead. AB - At the Department of Surgery of the Karl-Franzens University Hospital, a number of programs are available that use new technologies for training of students, interns and residents. The first project, the Virtual Medical Campus Graz (vmc.unigraz.at) is one of the "New Media in Teaching at Austrian Universities" projects. The general objectives of this initiative are innovation in teaching, intensification of contacts and easier access to education: The second project is virtual laparoscopy and surgical training. Unlike the traditional training modalities, computer-based training provides efficient and effective learning through an objective training tool that can assess learners' performance. It is to be expected that new media and technologies will continue to provide innovative approaches to learning and that the competition between new and traditional systems will continue to enliven the educational scene. PMID- 15747974 TI - Virtual reality in laparoscopic surgery. AB - Although the many advantages of laparoscopic surgery have made it an established technique, training in laparoscopic surgery posed problems not encountered in conventional surgical training. Virtual reality simulators open up new perspectives for training in laparoscopic surgery. Under realistic conditions in real time, trainees can tailor their sessions with the VR simulator to suit their needs and goals, and can repeat exercises as often as they wish. VR simulators reduce the number of experimental animals needed for training purposes and are suited to the pursuit of research in laparoscopic surgery. PMID- 15747975 TI - Use of information technology to improve quality of healthcare: Kosova's telemedicine project and international virtual e-hospital as an example. AB - The idea to create the Telemedicine Project of Kosova and the International Virtual e-Hospital Network of Kosova, was presented at G8-Meeting, in Berlin May 4-5, 2000, by the senior author (RL) then Assistant Professor of Surgery at Virginia Commonwealth University, in Richmond, Virginia during the presentation, the "The anatomy of war and destruction of Kosova: The alumni view on reconstruction of heath in Kosova". Following wide spread acceptance by many countries and institutions around the world of the idea of creating a virtual e hospital in Kosova,, became a reality. To ensure the creation of a telemedicine center and realization of the project in Kosova, the Telemedicine Association of Kosova was created in Prishtina, September 30, 2000. On February 2, 2001 a Memorandum of Understanding (MOU) for the implementation of TPK was signed between all stakeholders in health in Kosova. After extensive preparation, with the funding from the European Agency for Reconstruction, the Telemedicine Center of Kosova (TCK) was inaugurated on December 10, 2002, making way for the official beginning of the first phase of development of the TPK. This historical moment for Kosova and for the Balkan countries received extensive media coverage locally and abroad. Currently we are at the second year and phase of TPK realization. The Telemedicine Center of Kosova (TCK) is a state-of-the-art telemedicine center, the best in the Balkan and southeastern Europe. As such TCK is providing basic foundations for development of educational medical programs within the Kosova's medical system; as well it is establishing the standards of regional and international consultations and collaboration in the Balkan Peninsula. Furthermore, it is providing a solid basis for creation of human capacity that will lead and implement telemedicine program in the nine regional telemedicine centers of Kosova, for many years and decades to come. PMID- 15747976 TI - Telemedicine and mobile surgery in extreme conditions: the Ecuadorian experience. AB - Whenever we perform surgical procedures in modern operating rooms with state of the art technology, or attend patients in the comfort of our office, we should reflect on the fact that millions of people have no access to the most elemental surgical care. At the same time that mankind progresses, the gap between those who can benefit from the advances of science and the underprivileged societies, widens every day. We believe that surgeons have the obligation to strive for excellence in their practice; however, the benefits of their art and science should also extend those unfortunate populations in distant and remote areas. Therefore, efforts to develop means of reaching out to such countries and communities should also be the responsibility of conscientious physicians. PMID- 15747977 TI - Telemedicine in the Indian context: an overview. AB - One sixth of humanity accounting for 1100 million people live in India. India is generally considered to be a developing country and certainly there is considerable scope for improving the per capita income and the average standard of life. India however is a paradox. We now produce and launch our own satellites. Information is being gathered regarding the feasibility of launching a HEALTHSAT - a satellite exclusively for purposes of health care. There has been an unprecedented growth and development in Information Technology. Satellite transmission, fiber optic cables, increasing band width, fall in computer prices, licensing of private internet service providers, internet thro' cable etc have become the buzz words even in suburban and rural India. Theoretically, it is easier to set up an excellent telecommunication infrastructure in suburban and rural India, to increase the reach of the limited number of urban specialists, than to place hundreds of specialists in places devoid of specialists. Telemedicine therefore is the answer. It is universally acknowledged that Indians have made enormous contributions in the field of information technology. We no longer have to follow or piggy back. We now leap frog!! PMID- 15747978 TI - Telehealth: the backbone of healthcare financing. AB - Malaysia, like many other countries faces major challenges in meeting increasing demands with limited resources. Changes in demography, life-style diseases, increasing consumer expectations, new medical technologies and rapid economic growth all fuel demand for more healthcare services. There are problems related to the distribution and delivery of healthcare services, and there is inadequate integration of healthcare delivery and continuity of care is a major concern. Resources tend to be concentrated in the very expensive hospital sector, although services would be cost-effectively and conveniently delivered at primary care level. There is no ideal healthcare system, and how healthcare is supported and organized for service delivery influences the country's social, economic and political well-being. Like many other countries, Malaysia is undergoing health reform in meeting these challenges, and is becoming more reliant on telemedicine and telehealth. PMID- 15747979 TI - Telepresence and telemedicine in trauma and emergency care management. AB - The use of telemedicine is long-standing, but only recently has been applied to the specialties of trauma, emergency care, and surgery. Subsequently the concepts of teletrauma, telepresence, and telesurgery have evolved and are being integrated into modern care of trauma and surgical patients. This chapter will review the current applications and future endeavors of telemedicine and telepresence to trauma and emergency care as the new frontiers of telemedicine application. PMID- 15747980 TI - Telepresence and telementoring in surgery. AB - Telesurgical mentoring has evolved as an important subset of telemedicine, yet has remained an underutilized technique when all its potential is considered. As applied to surgery, telementoring is used when an experienced surgeon assists or directs another less experienced surgeon who is operating at a distance. Two- and three-dimensional, video-based laparoscopic procedures are an ideal platform for real-time transmission and thus for applying telementoring to surgery. The images viewed by the operating surgeon can easily be transmitted to a central "telesurgical mentor" and permit an intraoperative interaction. Several studies have demonstrated the practicality, effectiveness and safety of surgical telementoring. The goal of this application of telemedicine is to improve surgical education and training, expand patient care and improve health care delivery by allowing access to surgical specialists. Eventually, surgical telementoring could assist in the provision of surgical care to underserved areas and potentially facilitate the teaching of advanced surgical skills worldwide. PMID- 15747981 TI - Medical care from space: telemedicine. PMID- 15747983 TI - Creating a virtual sanctuary for scholars. AB - When people work together digitally, material can be added to a safe shared environment at anytime and can be accessed anywhere. The solutions generate new questions and new problems that then become interconnected. Cultivating this new and rapidly evolving technology of peer-to-peer platforms (1) may prove to be an extremely cost effective way to solve complex institutional or individual dilemmas. The solutions reflect the richness of two or more protected and engaged scholars or institutions working together with a refined degree of connection. After all, as Philip Condit, the CEO of the Boeing Company is often quoted as saying ... "none of us is as smart as all of us." The path to global connection hinges on relationships that are made and sustained one at a time. Progress together along the path similarly is made one step at a time. Sustained progress requires compelling reasons to do this in ways that are consonant with what it means to be human. PMID- 15747982 TI - The French language virtual medical university. AB - The work program of the French Language Virtual Medical University started about 2 years ago, and entered into a real active phase of development and implementation in 2002. Various national programs and organizational initiatives should accelerate and facilitate further appropriation of modem pedagogical approaches by knowledge providers and all users of the FSVMU, so as to advance on the way of virtual education. The French Language Virtual Medical University under the auspices of both the National Conference of Deans of Medical Schools and the International Conference of Deans of French-Speaking Medical Schools has been developed to create a major Internet portal for French-speaking distance medical learning and teaching. This construct should be representative of all medical schools in France and French-speaking countries. Contents will also be translated into English, Spanish and Arabic. All medical disciplines with their various levels of teaching are to be included. Cross-related fields are also going to be present in order to offer full range programs. The latter are intended to provide both initial and continuing education for medical students as well as all other categories of health professionals and medical and scientific research workers. To develop the appropriate technology and make such a portal, on one hand correspond to the specific educational requirements and proper training for health professionals, and on the other hand provide a general access to e-learning in all schools of medicine; in keeping with such goals, the following approaches should be stressed upon. To build a virtual space where individual patients, their families, patient associations as well as the general public, can obtain medical information of good quality for the purposes of both education and prevention. Providing such categories with reliable and validated sources of information, and offering an ethical basis for the increasing practice of e-medicine, represent in today's and tomorrow's world one of the great modern challenges for the medical community. Beyond the sole medical domain, the model offered by the FSVMU should be useful for the French-speaking Universities in all the fields of knowledge and training which are candidate to benefit from new educational means and methods. PMID- 15747984 TI - [Diabetes mellitus--definition, classification and diagnosis]. AB - Diabetes mellitus comprises of a group of heterogeneous disorders which have an increase in blood glucose concentrations in common. The current classifications for diabetes mellitus type 1-4 are described and the main features of type 1 and type 2 diabetes are compared to allow for better discrimination between these diabetes types. Furthermore, the criteria for the correct biochemical diagnosis during fasting and during oral glucose tolerance tests are summarized. These data form the basis of the recommendations of the Austrian Diabetes Association for the clinical practice in diabetes. PMID- 15747985 TI - [Type 2 diabetes mellitus--screening and prevention]. AB - The prevalence of diabetes is increasing in westernized countries. In addition, about half of all patients suffering from diabetes are not diagnosed. The current article represents the recommendations of the Austrian Diabetes Association for the screening and prevention of type 2 diabetes, based on currently available evidence. PMID- 15747986 TI - [Life-style modification]. AB - Diabetes education, nutritional advice, exercise recommendations and training for smoking cessation comprise the mainstay of diabetes treatment before starting drug therapy. Prevention as well as treatment of diabetes mellitus is positively influenced by life-style modification. In addition cardiovascular risk factors can be reduced by such measures. The article gives in brief recommendations leading towards life-style modification based on current evidence. PMID- 15747987 TI - [Therapy with oral antidiabetic drugs]. AB - Hyperglycemia contributes to morbidity and mortality in diabetic patients. Reaching treatment targets with regard to control of glycemia is thus a central goal in the treatment of diabetic patients. The present article represents the recommendations of the Austrian Diabetes Association for the practical use of oral antidiabetic drugs according to current scientific evidence. PMID- 15747988 TI - [Therapy with insulin]. AB - Hyperglycemia contributes to morbidity and mortality in diabetic patients. Reaching treatment targets with regard to control of glycemia is thus a central goal in the treatment of diabetic patients. The present article represents the recommendations of the Austrian Diabetes Association for the practical use of insulin according to current scientific evidence. PMID- 15747989 TI - [Hypertension]. AB - Hypertension is a highly prevalent comorbidity of diabetes mellitus. It contributes substantially to morbidity and mortality in those patients. The present article represents the recommendations of the Austrian Diabetes Association for the treatment of hypertension in diabetic patients according to current scientific evidence. PMID- 15747990 TI - [Lipids--diagnosis and therapy in type 2 diabetes]. AB - Hyper- and Dyslipidemia contribute to cardiovascular morbidity and mortality in diabetic patients. Pharmacological therapy with statins has convincingly proved to reduce cardiovascular risk in diabetic patients. The present article represents the recommendations of the Austrian Diabetes Association for the use of lipid-lowering drugs in diabetic patients according to current scientific evidence. PMID- 15747991 TI - [Inhibition of platelet aggregation]. AB - Acute atherothrombotic complications, as part of the accelerated atherosclerosis, contribute to cardiovascular morbidity and mortality in diabetic patients. Inhibition of platelet aggregation can reduce the risk for acute atherothrombosis. The present article represents the recommendations of the Austrian Diabetes Association for the use of antiplatelet drugs in diabetic patients according to current scientific evidence. PMID- 15747992 TI - [Diabetic neuropathy]. AB - These are the guidelines for diagnosis and treatment of diabetic neuropathy. This diabetic late complication comprises a number of mono- and polyneuropathies, plexopathies, radiculopathies and autonomic neuropathy. The position statement summarizes characteristic clinical symptoms and techniques for diagnostic assessment of diabetic neuropathy. Recommendations for the therapeutic management of diabetic neuropathy, especially for the control of pain in sensorimotoric neuropathy, are provided. PMID- 15747993 TI - [The diabetic foot]. AB - These are the guidelines for preventive care, diagnosis and treatment of diabetic foot syndrome. Diabetic neuropathy, peripheral vascular disease, bone deformity and altered biomechanics are foot-related risk conditions. The position statement is focused on screening methods and recommendations for clinical care for diabetics, who currently have no foot ulcers. A decision pathway is offered with respect to diagnosis and management of diabetic patients at increased risk or who manifest injuries. PMID- 15747994 TI - [Diabetic nephropathy]. AB - Diabetes mellitus is the leading single cause for renal replacement therapy. Its development and progression, however, can be ameliorated by adequate therapy. The present article represents the recommendations of the Austrian Diabetes Association for the prevention and treatment of diabetic nephropathy. PMID- 15747995 TI - [Diagnosis, therapy and follow-up of diabetic eye diseases]. AB - Diabetes mellitus causes diabetic retinopathy and maculopathy, optical nerve neuropathy, cataract and defects of the eye muscles. The incidence of these defects correlates with duration and quality of the metabolic control. The recommendations of the Austrian Diabetes Association for the diagnosis, the therapeutic measures and requirements for adequate follow-up depending on the stages of the different forms of diabetic eye diseases are summarized. PMID- 15747996 TI - [Gestational diabetes mellitus]. AB - Gestational diabetes (GDM) is defined as any degree of glucose intolerance with onset or first recognition during pregnancy and is associated with increased feto maternal morbidity as well as long-term complications in mothers and offspring. GDM is diagnosed by an oral glucose tolerance test (OGTT) or fasting glucose concentrations in the diabetic range. In case of a high risk for GDM/type 2 diabetes (history of GDM or prediabetes [impaired fasting glucose or impaired glucose tolerance]; malformation, stillbirth, successive abortions or birth weight > 4500 g in previous pregnancies) performance of the OGTT (120 min; 75 g glucose) is recommended already in the first trimester and--if normal--the OGTT should be repeated in the second/third trimester. In case of clinical symptoms of diabetes (glucosuria, macrosomia) the test has to be performed immediately. All other women should undergo a diagnostic test between 24 and 28 gestational weeks. If fasting plasma glucose exceeds 95 mg/dl, 1 h 180 mg/dl and 2 hrs 155 mg/dl after glucose loading (OGTT) the woman is classified as GDM (one pathological value is sufficient). In this case a strict metabolic control is mandatory. All women should receive nutritional counseling and be instructed in blood glucose self-monitoring. If blood glucose levels cannot be maintained in the normal range (fasting < 95 mg/dl and 1 h after meals < 130 mg/dl) insulin therapy should be initiated. Maternal and fetal monitoring is required in order to minimize maternal and fetal/neonatal morbidity and perinatal mortality. After delivery all women with GDM have to be reevaluated as to their glucose tolerance by a 75 g OGTT (WHO criteria). PMID- 15747997 TI - [Research methods in dentistry. 8. Methods for longitudinally detecting differences in bone density: digital subtraction radiography]. AB - Subtraction radiography has been developed in 1934 by the Dutch radiologist B. Ziedses des Plantes. The technique is based on the subtraction of two radiographic images. As a result, only the differences remain visible, and structures which have been unaltered in between both exposures, are suppressed. The technique was introduced also in dental radiology in the eighties in the last century. Since then, many studies have been performed showing the benefit of this technique in dental diagnosis. A requirement for reliable results is that the two radiographs which are used for the subtraction have equal brightness and contrast, as well as a completely identical projective geometry. Since radiographic images can be acquired digitally, methods have been developed to make density and contrast equal for both images used for the subtraction process even after the images have been taken, using dedicated software programs. The projection geometry of two images can be made identical in this way as well. Because of this, the technique can also be applied in general dental practice. PMID- 15747998 TI - [Premalignant lesions of the oral mucosa. Prognosis, treatment and follow-up]. AB - To consider the value of prognostic factors in the development of a squamous cell carcinoma from a leukoplakia of the oral mucosa, a retrospective study was performed. Clinical and histological data of 104 patients with oral leukoplakia were analyzed. Leukoplastic lesions with dysplasia in the initial biopsy (n = 38) had been treated by excision (n = 28), by laser evaporation (n = 6) or a combination of these treatments (n = 4). Non-dysplastic lesions (n = 66) had been excised (n = 48), evaporised (n = 17) or treated by excision as well as laser evaporation (n = 1). During follow-up of maximal 6 years (mean 3.6 years), 12 patients had developed an infiltrative squamous cell carcinoma at the site of the primary lesion, 2 within a period of 24 months. No relation could be found between on the one hand size (p > 0.2), clinical aspect (p > 0.2), location (p > 0.45), and primary treatment (p > 0.15) of the lesion, and on the other hand the risk of developing a squamous cell carcinoma. Only a relation could be found between (the intensity of) dysplasia and the development of a squamous cell carcinoma (p < 0.001). It was concluded that because of the high risk of developing a squamous cell carcinoma, patients with a dysplastic mucosal oral lesion should be followed during a prolonged time. PMID- 15748000 TI - [Dissertations 25 years after date 4. Periodontal surgery an important component of periodontal treatment]. AB - The aim of periodontal treatment is to make and keep the periodontal tissues healthy. A proper diagnosis including the assessment of the different risk factors is essential before successful treatment can be carried out. Oral hygiene instruction together with supra- and subgingival cleaning is a basic part of the treatment. Antibiotic treatment and smoking cessation programs also contribute to a good clinical result. Until the seventies, when almost nothing was known about riskfactors and antibiotic therapy, no study was performed to assess the long term effect of periodontal surgery being part of the treatment. Therefore a study was performed to assess clinically, radiographically and histologically the effect of periodontal surgery and oral hygiene on artificial periodontal defects. The results were reported in the thesis 'Surgery and oral hygiene on artificial periodontal defects in beagle dogs'. This study established that periodontal surgery can contribute to a healthy periodontium. A good oral hygiene is a prerequisite. These results have been confirmed by a long-term study in private practice. Pocketdepths of 5 mm and more after initial treatment is a good criterion in the decision making for periodontal surgery. PMID- 15748001 TI - [A painful swelling of the tongue]. AB - A 45-year-old patient had a progressive ulcer of the tongue. Routine examination, including a biopsy was not conclusive and could not entirely rule out a malignancy. The final diagnosis was made after generalised lesions developed, after which specific serological tests confirmed the diagnosis 'syphilis'. The patient was treated with benzathine benzylpenicillin and his complaints disappeared. In view of the increasing incidence of this disease remains an important consideration in case of ulcerative or other lesions in the oral cavity. PMID- 15748002 TI - [From a hippopotamus tooth to a porcelain tooth]. AB - The history of dentistry is written as if thousands of years of obscurity were followed by a couple of hundred years of enlightenment. The turning point was the work of the Father of Modern Dentistry Pierre Fauchard (1678-1761). But historically speaking this notion of progress has theoretical and practical disadvantages. In this article a practical example is given, that of porcelain. The production developed completely in the times of obscurity and only a simple pharmacist just saw the possibilities of the result in dentistry. PMID- 15748003 TI - Grant aims to decrease State's uninsured. PMID- 15748004 TI - Nurses reaching out to the working uninsured: one clinic's experience. PMID- 15748005 TI - Perceived barriers to adherence as described by individuals with HIV disease. PMID- 15748007 TI - Examining breastfeeding practices among low-income black women. PMID- 15748006 TI - Spirituality as a component in nursing care. AB - Narayanasamy's (2002) study supported spiritual coping mechanisms as an important part of coping and recovery in chronically ill patients. The results of this study can be used to support a research utilization project to teach nurses the importance of supporting and promoting patient spirituality as a component of nursing care. A feasibility issue would include the need for spiritual assessment tools. Future research could focus on the importance of spirituality in patients of different age groups. PMID- 15748008 TI - Demystifying bioterrorism: misinformation and misperceptions. AB - The true threat of bioterrorism remains mysterious and elusive to the common citizen. It principally has become the dominion of a few "experts", many of whom have limited apparent expertise, who have failed to effectively communicate the risks and realities to society, and have instead created an air of uncertainty surrounding the topic. Unlike the great classic deceptions of modern life (e.g., "the check is in the mail"), the misinformation and misperceptions associated with bioterrorism can be dangerous and are not merely humorous. Indeed, it is possible to grasp the facts as well as fallacies associated with bioterrorism, and, as a result, demystify this nightmare scenario and prepare for the "unthinkable". PMID- 15748009 TI - Predictors of emotional adjustment following traumatic injury: personal, social, and material resources. AB - INTRODUCTION: The identification of factors influencing emotional adjustment after injury may elucidate the design of assessment and treatment procedures in emergency medicine settings and suggest targets for early intervention to prevent the later development of psychological impairment. Personal, social, and material resources may be influential factors and require further evaluation. HYPOTHESES: Greater experiential avoidance, social constraints on discussing the trauma experience, and loss of material resources would be associated with more of the symptoms of post-traumatic stress and depression following traumatic injury. METHODS: Participants (n = 47) at a mean of 7.4 months post-injury, completed a telephone interview assessment, including evaluation of sociodemographic characteristics, conservation of resources, social constraints, acceptance and commitment, and symptoms of post-traumatic stress disorder (PTSD) and depression. Hypotheses were tested using multivariate regression analyses. RESULTS: Only greater social constraints were uniquely predictive of greater PTSD symptomatology. Higher levels of experiential avoidance, social constraints, and loss of material resources all were associated with greater levels of depression. CONCLUSION: Assessment of personal coping style, receptivity of social network, and loss of potential material resources following traumatic injury may facilitate identification of individuals at-risk for poorer post-injury adaptation. Psychosocial interventions targeting such individuals may be promising. PMID- 15748010 TI - The incident command system in disasters: evaluation methods for a hospital-based exercise. AB - OBJECTIVES: No universally accepted methods for objective evaluation of the function of the Incident Command System (ICS) in disaster exercises currently exist. An ICS evaluation method for disaster simulations was derived and piloted. METHODS: A comprehensive variable list for ICS function was created and four distinct ICS evaluation methods (quantitative and qualitative) were derived and piloted prospectively during an exercise. Delay times for key provider-victim interactions were recorded through a system of data collection using participant- and observer-based instruments. Two different post-exercise surveys (commanders, other participants) were used to assess knowledge and perceptions of assigned roles, organization, and communications. Direct observation by trained observers and a structured debriefing session also were employed. RESULTS: A total of 45 volunteers participated in the exercise that included 20 mock victims. First, mean, and last victim delay times (from exercise initiation) were 2.1, 4.0, and 9.3 minutes (min) until triage, and 5.2, 11.9, and 22.0 min for scene evacuation, respectively. First, mean, and last victim delay times to definitive treatment were 6.0, 14.5, and 25.0 min. Mean time to triage (and range) for scene Zones I (nearest entrance), II (intermediate) and III (ground zero) were 2.9 (2.0-4.0), 4.1 (3.0-5.0) and 5.2 (3.0-9.0) min, respectively. The lowest acuity level (Green) victims had the shortest mean times for triage (3.5 min), evacuation (4.0 min), and treatment (10.0 min) while the highest acuity level (Red) victims had the longest mean times for all measures; patterns consistent with independent rather than ICS-directed rescuer activities. Specific ICS problem areas were identified. CONCLUSIONS: A structured, objective, quantitative evaluation of ICS function can identify deficiencies that can become the focus for subsequent improvement efforts. PMID- 15748011 TI - Public hospital preparations for SARS outbreak: experience of Alexandra Hospital. AB - Alexandra Hospital (AH) was one of the public hospitals in Singapore that responded to the severe acute respiratory syndrome (SARS) crisis. Being the only public hospital that remained "SARS-free", i.e., with no documented intra institutional spread of disease, AH had to tackle a sudden, two-fold surge in hospital attendances and patient volume. Being the oldest hospital with a traditional open ward design and lack of proper isolation facilities, tough command and control policies had to be implemented to reduce the risk of a SARS outbreak. Stringent infection control measures, screening and triage, clinical procedures, and administrative policies all were important factors in helping the hospital balance the need to run routine operations while "fighting" SARS. Staff and people management also were crucial in keeping the workforce healthy and maintaining their morale and confidence during this difficult period. PMID- 15748012 TI - Current status of International Emergency Medicine fellowships in the United States. AB - OBJECTIVE: A consensus panel of Emergency Physicians with experience in international health has published a recommended curriculum for a formal fellowship in International Emergency Medicine. This article reviews the current International Emergency. Medicine (IEM) fellowships available to residency trained Emergency Physicians in the United States. METHODS: Every allopathic Emergency Medicine (EM) residency program in the United States was contacted via e-mail or telephone. Programs that reported having an IEM fellowship were asked detailed information about their program, including: (1) the number of years the program has been offered; (2) the duration of the program; (3) the number of fellows taken each year; (4) the number of fellowship graduates from each program and their current practice patterns; (5) how the fellowship is funded; and (6) whether a Masters Degree in Public Health (MPH) is offered. RESULTS: All 127 allopathic EM residency programs responded. Eight (6.8%) of these programs offered IEM fellowships. Of a total of 29 graduates identified, 23 (79.3%) were employed in academic medicine. All of the fellowships offered formal public health training and were funded by a combination of clinical billing and project specific grants and scholarships. All IEM fellowships described a curriculum that reflected the previously published recommendations. CONCLUSION: Opportunities in formal training in international health are increasing for graduates of EM residencies in the United States. The proposed curriculum for IEM fellowships seems to have been implemented and graduates of IEM fellowships seem to be applying their training in international projects. PMID- 15748013 TI - Successful development and implementation of' a tactical emergency medical technician training program for United States federal agents. AB - INTRODUCTION: The emerging need for tactical operations in law enforcement often places personnel involved at risk. Tactical operations often are carried out in environments in which access to emergency care is limited. With the war against terrorism expanding, special operations involving United States federal agents are occurring worldwide. Currently, there are very few tactical medicine curricula training traditional emergency medical services (EMS) providers to operate in these high-risk missions. Trainees in existing programs must have previous EMS experience, and are selected from a wide range of backgrounds. The goal of this study is to examine a Special Agent Emergency Medical Technician (SAEMT) training curriculum developed specifically for federal special agents with prior experience in tactical operations, but without previous medical training. METHODS: An analysis of the SAEMT Program given to federal agents of the Drug Enforcement Administration (DEA) in Quantico, Virginia between July 2000 and April 2002 was performed. The SAEMT curriculum provided enrolled agents 181.5 hours of training in tactical emergency topics, including medical mission planning, logistics, operations, evacuation, and weapons training. In addition, SAEMT concurrently provides emergency medical technician (EMT) training. All of the participants were DEA agents with no previous medical training. Upon completion of the course, all participants took the National Registry of EMT Basic examination. Measured endpoints included course completion rate and performance on certifying examinations. RESULTS: Ninety-five agents were enrolled and successfully completed the SAEMT course between July 2000 and April 2002. Of the agents enrolled, 84 (88%) passed the National Registry of EMTs-Basic examination within two attempts. CONCLUSION: The SAEMT Program provides basic emergency medical training to federal special agents with no previous medical experience. The design of this program provides a useful template to meet the expanding demand for tactical emergency medical personnel. PMID- 15748014 TI - Triage decisions of United Kingdom police firearms officers using a multiple casualty scenario paper exercise. AB - INTRODUCTION: British police officers authorized to carry firearms may need to make judgments about the severity of injury of individuals or the relative priority of clinical need of a group of injured patients in tactical and non tactical situations. Most of these officers receive little or no medical training beyond basic first aid to enable them to make these clinical decisions. Therefore, the aim of this study is to determine the accuracy of triage decision making of firearms-trained police officers with and without printed decision support materials. METHODS: Eighty-two police firearms officers attending a tactical medicine course (FASTAid) were recruited to the study. Data were collected using a paper-based triage exercise that contained brief, clinical details of 20 adults and 10 children. Subjects were asked to assign a clinical priority of immediate or priority 1 (P1); urgent or priority 2 (P2); delayed or priority 3 (P3); or dead, to each casualty. Then, they were provided with decision-making materials, but were not given any instruction as to how these materials should be used. Subjects then completed a second triage exercise, identical to the first, except this time using the decision-support materials. Data were analyzed using mixed between-within subjects analysis of variance. This allowed comparisons to be made between the scores for Exercise 1 (no decision support material) and Exercise 2 (with decision-support material). It also allowed any differences between those students with previous triage training and those without previous training to be explored. RESULTS: The use of triage decision-making materials resulted in a significant increase in correct responses (p < 0.001). Improvement in accuracy appears to result mainly from a reduction in the extent of under-triage. There were significant differences (p < 0.05) between those who had received previous triage training and those who had not, with those having received triage training doing slightly better. CONCLUSION: It appears that significant improvements in the accuracy of triage decision-making by police firearms officers can be achieved with the use of appropriate triage decision support materials. Training may offer additional improvements in accuracy, but this improvement is likely to be small when decision-support materials are provided. With basic clinical skills and appropriate decision-support materials, it is likely that the police officer can make accurate triage decisions in a multiple-casualty scenario or make judgments of the severity of injury of a given individual in both tactical and non-tactical situations. PMID- 15748016 TI - Information technology and public health management of disasters--a model for South Asian countries. AB - This paper highlights the use of information technology (IT) in disaster management and public health management of disasters. Effective health response to disasters will depend on three important lines of action: (1) disaster preparedness; (2) emergency relief; and (3) management of disasters. This is facilitated by the presence of modern communication and space technology, especially the Internet and remote sensing satellites. This has made the use of databases, knowledge bases, geographic information systems (GIS), management information systems (MIS), information transfer, and online connectivity possible in the area of disaster management and medicine. This paper suggests a conceptual model called, "The Model for Public Health Management of Disasters for South Asia". This Model visualizes the use of IT in the public health management of disasters by setting up the Health and Disaster Information Network and Internet Community Centers, which will facilitate cooperation among all those in the areas of disaster management and emergency medicine. The suggested infrastructure would benefit the governments, non-government organizations, and institutions working in the areas of disaster and emergency medicine, professionals, the community, and all others associated with disaster management and emergency medicine. The creation of such an infrastructure will enable the rapid transfer of information, data, knowledge, and online connectivity from top officials to the grassroots organizations, and also among these countries regionally. This Model may be debated, modified, and tested further in the field to suit the national and local conditions. It is hoped that this exercise will result in a viable and practical model for use in public health management of disasters by South Asian countries. PMID- 15748015 TI - Effects of prehospital spinal immobilization: a systematic review of randomized trials on healthy subjects. AB - OBJECTIVE: To evaluate the effects of spinal immobilization on healthy participants. METHODS: A systematic review of randomized, controlled trials of spinal immobilization on healthy participants. RESULTS: Seventeen randomized, controlled trials compared different types of immobilization devices, including collars, backboards, splints, and body strapping. For immobilization efficacy, collars, spine boards, vacuum splints, and abdominal/torso strapping provided a significant reduction in spinal movement. Adverse effects of spinal immobilization included a significant increase in respiratory effort, skin ischemia, pain, and discomfort. CONCLUSIONS: Data from this review provide the best available evidence to support the well-recognized efficacy and potential adverse effects of spinal immobilization. However, comparisons of different immobilization strategies on trauma victims must be considered in order to establish an evidence base for this practice. PMID- 15748017 TI - Disasters and development: Part 2: Understanding and exploiting disaster development linkages. AB - This lesson is a continuation of Disasters and Development: Part 2: Understanding and Exploiting Disaster-Development Linkages published in Prehospital and Disaster Medicine in Volume 17, Number 3. It identifies the goals of a specific damage mitigation project that can be incorporated into a regular development project and the mechanisms for obtaining the mitigation component of such a project. Mechanisms for assessing the success of such a project are discussed. It stresses the importance of the application of building codes, associated training programs, and more extensive use of zoning regulations in urban development that decrease the population at risk and the likelihood of damage to industrial facilities. Disasters can elevate the development potential of a society at risk for damage from a hazard. The political impact of damage and disruption can be a catalyst for change. Development opportunities often are compromised because of an excessive focus on relief assistance. Interventions designed to mitigate the damage from a given hazard are particularly effective when they focus on areas at particularly high risk for actualization of the hazard. Support from the private sector, including the non-formal sector, is a key element of successful reconstruction management. The period of recovery is an opportunity for general assistance to government with administrative procedures, including enhanced management training programs. PMID- 15748018 TI - Disasters and development: Part 3: Assessing trade-offs in investing in vulnerability reduction. AB - This lesson describes how a government decides whether and how much it should spend on vulnerability reduction. There are techniques and methods by which decision-makers compare development alternatives. The differences between the risk that a potentially catastrophic event will occur and uncertainty are described, with uncertainty providing greater difficulty in economic analyses. There is a range of methods for identifying the complex mix of competing costs and benefits associated with any restructuring of investment priorities to accomplish disaster mitigation. The possibilities are described in terms of the opportunity costs and present value. Impact and consequent losses include: (1) direct monetary effects; (2) indirect monetary effects; (3) direct, non-monetary effects; (4) indirect, non-monetary effects; and (5) loss of non-renewable natural resources. The difficulties in assigning values to these effects are described, as well as the means of judging the cost-effectiveness of such interventions. An advantage of screening projects using a framework of analytical methods is that it can assist in focusing on a variety of possible outcomes and make the factors influencing these outcomes quite explicit. PMID- 15748019 TI - [Immunopathological and histopathological aspects of ectopic implantation with possible effect on fertility disorders]. AB - OBJECTIVE: To evaluate leucocytic infiltration of fetomaternal interface in ectopic pregnancy and to evaluate the changes in cell immunity against trophoblast (AT-CMI) in women with extrauterine pregnancy (GEU) in their medical history. To assess the effect of these factors on possible fertility disorders in a woman. DESIGN: A retrospective study. SETTING: Mother and Child Care Institute, Prague. METHODS: In most of the patients, we addressed GEU through laparoscopy. The tube was extirpated in toto and immediately fixed in Baker's solution. Thereafter, it was prepared in a dissection microscope and then processed in a standard way. In order to identify the intensity of AT-CMI, we used the leucocyte migration inhibition test. The cytotrophoblastic cell line JAR was used as an antigen. The degree of inhibition of the migration was monitored by means of a computer image analyser. Inhibition of migration below 75% was rated as favourable. RESULTS: We monitored the presence of inflammatory infiltrate in the place of implantation and correlated the findings with the hCG levels and the presence of the foetal ovum or its part in the tube. In 28 patients (23.5%) of the total number of 119 patients in the group, we observed an inflammatory infiltrate in the place of implantation. In these patients, the hCG levels were lower and in 17 of them (60.7%) we did not prove the presence of a foetal ovum or its parts. In women with GEU in their medical history, the AT-CMI positivity was established in 61.1% of the women 1 year after surgery, in 56.8% of the women 1-3 years after surgery and in 41.2% of the women 3 years after surgery. CONCLUSION: Ectopic pregnancy involves a pathological fetomaternal interface. The leucocytic infiltrate in the area of implantation may be of secondary character and may cause gradual destruction of the ectopically positioned product of conception. The results of our study indicate a possible participation of the increased AT CMI in the destruction of the ectopically located trophoblast. Persisting anti trophoblast immunity may influence the occurrence and course of further gravidities. PMID- 15748020 TI - [Malignant trophoblastic tumors (MTT) treated in the years 1955-2004 in trophoblastic disease center in the Czech Republic (TDC-CZ): clinical pathological features, curability, typing, pathogenesis]. AB - OBJECTIVE: Clinical- pathological features, typing, curability and pathogenesis of malignant tumors of trophoblast (MTT). DESIGN: A retrospective analysis. SETTING: Trophoblastic Disease Center in the Czech Republic (TDC-CZ), Mother and Child Care Institute, 3rd Medical Faculty, Charles University, Prague. METHODS: Revision of 379 MITT cases treated at TDC-CZ and comparison of their histological picture with developmental stages of orthologic trophoblast from the standpoint of MTT typing and pathogenesis. The determination of curability of different histological types and risk stages (RS) used in TDC-CZ and a comparison with RS axccording to FIGO, WHO and NIH. Differentiation of undifferentiated choriocarcinoma (CH-Und) alias Epitheloid Trophoblastic Tumor (ETT) from the given cohort of MIT and establishment of its clinical and biological properties, curability and formal pathogenesis. RESULTS: Three hundred and seventy nine MTT cases were classified into 5 histological types onthe basis of analogy with developmental stages of 7 to 14 days old trophoblast. 1: typical "classical" choriocarcinoma - No Special Type (CH-NST), which forms more than 80% of MTT. Moreover, the degree of differentiation of tumorous trophoblast and its prevailing (cytological) type made it possible to define other 4 types, i.e.: 2: CH-syncytio-trophoblastic (CH-Syn), representing 3.8%; 3: CH-cytotrophoblastic CH Cyt with 3.3%; 4: CH-dissociated (CH-Dis), representing 6%; 5: CH undifferentiated (CH-Und) with 6.8%. Mortality due to MIT of all mentioned types reached 94% until 1963, decreased to 43% until 1980 and has been 5.8% in the period of 1981-2004. In the latter period of time (1981-2004), mortality due to CH-Cyt proved to be 40%, that due to CH-Dis being 11%, and CH-Und 18%, though. Mortality of s.c. Placental Site Trophoblastic Tumor, which includes our CH-Cyt and CH-Dis therefore forms 21.4%. We have been using four RS in TDC-CZ. The following outline includes only the main features: 1st RT includes CH-NST < 30mm limited to uterus in connection with mole. 2nd RS includes CH-NST > 30mm after birth. 3rd RS includes CH-NST with multiple metastases outside GIT and CNS and MTT with the CH-Cyt, Dis, Und component < 75%. 4th RS includes CH-NST with metastases in CNS or GIT. MTT with CH-Cyt Dis, Ned component < 75% with metasteses and MTT with the same components > 75% In the last 23 years 1st RS and 2nd RS includes 85 % of all MTT in the TDC-CZ and curability is 100%. In the 3rd RS curability decreases to 64.3% and decreases to 55.6% in the 4th RS. According to FIGO classification the 1st RS forms 48%, 2nd RT represents 17% and 100% curability applies for both of them. 3rd a RS includes 20% of 100% curability, 3rd bc RS forms 10% with 67% curability and 4th abc RS includes 5 % with 50% curability. In using the WHO classification with four RS, their percentage representation is similar to our classification with similar curability; nevertheless the 1st RS and 2nd RS did not detect almost 8% of MTT, which ended with exitus. 3rd RS according to FIGO is overestimated in view of 100% curability and the abc degree in 1st and 2nd RS are only of theoretical significance and irrelevant for the choice of treatment. The closest results comparable with our classification were those of NIH. A very careful clinical-pathological analysis of 25 CH-Und-ETT, detected among 379 MTT revealed that CH-Und-ETT is anaggressive malignant form of CH, which is best derived from undifferentiated 7-8 days old trophoblast. It is insidious for its seemingly primary extragenital symptomatology in seven out of 25 cases, low hCG values and poor sensitivity to chemotherapy. CONCLUSION: 1) The comparison of histological pictures of 379 MTT with developmental stages of orthologic trophoblast of 7-14 days old embryo was the basis for classification of 5 types of choriocarcinoma (CH): 1. Differentiated CH "No Special Type" (CH-NST), 2. Syncytiotrophoblastic CH (CH Syn), 3. Cytotrophoblastic CH (CH-Cyt), 4. Dissociated CH (CH-Dis), and 5. Non differentiated CH (CH-Und); 2) We have determined their percentual (and absolute) occurrence in the group of 379 MTT treated in CTN in the years 1955-2004. 3) We have described biological properties of individual types of CH and established the way they influence curability. 4) Four degrees of risk (RS) were specified in relation to 7 types of risk factors observed (1. size of tumor, 2. type of preceding pregnancy, 3. interval from pregnancy to the diagnosis, 4. histological type of CH, 5. number of metastases, 6. localization of metastases, 7. values of hCG). It has become obvious how RS influenced curability of CH (1st and 2nd RS forms 85% of all CH's and their curability is 100% (!), 3rd and 4th RS are represented in 15% and their curability is 64% in the 3rd RS and 55% in the 4th RS. 6) The curability reached in CTN was compared with that determined according to FIGO, WHO and NIH, respectively. The results proved to be similar, but in case of FIGO the 3rd degree was overestimated and the degrees abc in the 1st and 2nd RS were of theoretical importance only, therefore being of no values for the choice of treatment. Low and medium score according to WHO did not detect 8% of women who had died. The CH curability according to RS, having been recommended by NIH and used in the American Centers was virtually the same as our results. 7) It has been proved that the histological type of CH significantly influenced the determination of RS in the given patient. 8) CH-Und-ETT represents the least differentiated form of MTT, in other words choriocarcinoma. This is associated with a low production of HCG, mostly between 10(1) and 10(3) mIU/ml. 9) Pathogenesis of CH-Und ETT-ETT we derive from the earliest, undifferentiated stage of orthological trophoblast. The origin from the differentiated intermediate trophoblast chorion leeve we considerei improbable. 10) There are continuous transitions from CH-Und-ETT and PSTT to CH-NST, representing an analogy to grading in other malignant epithelial tumors. PMID- 15748021 TI - [Immunologically conditioned fertility disorders in men--experience of the immunobiological department of the Institute for Maternal and Child Care]. AB - OBJECTIVE: To provide an overview of the results obtained from the more than ten year systematic monitoring of the contribution of immunopathological mechanisms to the ethiopathogenesis of fertility disorders in men. DESIGN: A summarising retrospective study. SETTING: Mother and Child Care Institute, Prague. METHODS: The data source is a selected group of a total of 3,800 couples, who were examined in the Immunobiological Department (ID) of the Mother and Child Care Institute (MCCI) in Prague- -Podoli for fertility disorders in the past. From among the laboratory quantities, the following ones were systematically monitored: a) sperm parameters; b) cell-mediated immunity against spermatozoa; c) the presence of anti-spermatozoa auto-antibodies attached to spermatozoa; d) in a part of the clinical group, the serum concentrations of FSH, testosterone, SHBG and the free androgen index (FAI) were monitored. RESULTS: After selection was made according to defined criteria, the group comprised of a total of 1,680 men, of whom 49.4% were normozoospermic and 50.6% suffered from some form of seminal pathology. Increased cell-mediated immunity against spermatozoa was identified in 10.2% of fertile men, in 18.5% of normozoospermic men and in 66.3% of azoospermic men. In asthenozoospermic and teratozoospermic men, the increased cell-mediated immunity against spermatozoa was identified in 48.3% and 53.1% of them, respectively. The auto-antibodies attached to spermatozoa were identified in 3 out of every 42 fertile men (7%), while in asthenozoospermic men, it was a total of 21% (IgA antibodies) and 22% (IgG antibodies). As concerns the concentration of free androgens (FAI) in the serum, there was no difference among the individual subgroups of men. In oligoasthenozoospermic men, FSH was significantly higher on the average in comparison with normospermic men. CONCLUSION: The significantly higher incidence of increased cell-mediated immunity against spermatozoa in men with a pathological spermiogram in comparison with the control group (fertile men and normozoospermic men) indicates that cell-mediated immunity participates in the pathogenesis of seminal pathologies. PMID- 15748022 TI - [Possibilities of using 3D ultrasonography in imaging of lower urinary pathways]. AB - OBJECTIVE: To draw attention to the possibilities of using 3D-ultrasonography in imaging lower urinary pathways and vagina. DESIGN: A pilot study. SETTING: Mother and Child, Care Institute Prague. METHODS: the examinations were performed in patients within the framework of a complex urogynecological examination following the urodynamic examination. The 3D analysis and reconstruction was performed in asymptomatic nulliparas, in patients with primary stress incontinence and also in women with relapse of stress incontinence. All reconstructions were performed by the author. CONCLUSIONS: 3D ultrasonography represents a noninvasive imaging method which, in indicated cases, may be suitable for completion of 2D ultrasound examination, representing the gold standard of lower urinary pathways imaging. By no means is the 3D ultrasonography a method competing with 2D ultrasonography. Both methods could suitably supplement each other especially in patients with relapsing stress incontinence after urogynecological operations performed. PMID- 15748023 TI - [Complications of the invasive prenatal diagnostic methods]. AB - OBJECTIVE: To provide a literary overview of the complications associated with the invasive prenatal diagnostic methods in the 1st and 2nd trimester of pregnancy. DESIGN: A summarising article. SETTING: Mother and Child Care Institute, Prague. METHODS: Compilation of data from professional literature. RESULTS AND CONCLUSION: The global trend is characterised by the effort to move prenatal diagnostics into the period of the first trimester. The invasive diagnostic methods of this period - sampling of chorionic villi and timely amniocentesis are subjects of a number of discussions, which deal primarily with their reliability and safety. The main indicator is the evaluation of foetal losses occurring in connection with the action. So far, the safest method is second-trimester amniocentesis; for early diagnostics, the trans-abdominal sampling of chorionic villi is preferred to transcervical one and to early amniocentesis. PMID- 15748024 TI - [Incidence of birth defects in children of mothers using medications in the 1st trimester of pregnancy in the Czech Republic 1996-2002]. AB - OBJECTIVE: To process the results on the incidence of birth defects in the children of mothers using medicines in the first trimester of pregnancy in the Czech Republic during 1996-2002. DESIGN: A retrospective analysis of data from the National Register of birth Defects in the Czech Republic. METHODS: The data from the nationwide register of birth defects administrated in the Institute of Health Information and Statistics of the Czech Republic for the period 1996-2002 were used for the study. The data on the incidences of selected types of birth defects identified in children born to mothers using medicines in the first trimester of pregnancy were compiled and an analysis of these medicines was carried out in relation to the diagnosed types of birth defects. The data on healthy children born to mothers using medicines during the 1st trimester of pregnancy were used as the control sample. RESULTS: In our retrospective analysis covering the years 1996-2002, a total of 1,146 birth defects were identified in children of women using medication in the 1st trimester of pregnancy and a total of 891 children affected in this way were identified. CONCLUSION: The study presents the updated results of these analyses based on nationwide registration of birth defects for a seven-year period. It analyses the types of birth defects in the children of women using medication in the 1st trimester of pregnancy and the types of medicines used by these women. In our study, we have found a significantly higher incidence in the population of the children born to women using medicines in the 1st trimester of pregnancy in several cases. From among the group of cleft defects affecting the central nervous system, this concerns for example anencephalia and spina bifida, as well as congenital hydrocephalus, anophthalmy/microphthalmy, auricle abnormalities and limb reduction deformities. From these results that we produced, it is of course not possible to draw significant conclusions with a clinical impact. However, pregnant women should avoid any medicines in the first trimester of pregnancy, except for those cases, which are strictly indicated from the medical point of view and which are recognised as reasonably safe. As concerns the use of other medicines, their genetic and teratogenic risk should be assessed comprehensively by a specialised clinical geneticist on a case-by-case basis. PMID- 15748026 TI - [Survival of children born with selected types of birth defects in Czech Republic]. AB - OBJECTIVE: A retrospective study of data with an analysis of the prenatal and postnatal incidence of selected types of birth defects in the Czech Republic during the period from 1994 to 2003. The objective was also to analyse the percentage of deceased children born with selected types of congenital defects from the total number of deceased children. DATA SOURCE AND METHODS: The data on prenatal diagnostics were obtained from particular department of medical genetics in the Czech Republic. The data on the incidences of birth defects in the born children were obtained from the data of the National Health Registers, the administration of which has been entrusted to the Institute of Health Information and Statistics of the Czech Republic (IHIS CR). Specifically, the data from the National Register of Congenital Defects and the National Register of Neonates were used. The analysis of the frequencies of prenatally and postnatally diagnosed cases of twelve selected types of birth defects was carried out for the Czech Republic for the period from 1994 to 2003. For the cases of postnatally diagnosed defects, an analysis of survival and mortality of the affected individuals during the 1st year of life was carried out for selected types of birth defects. RESULTS: In the period 1994-2003, a total of 932,153 children were born in the Czech Republic. Including the cases of successful prenatal diagnostics, we analysed 268 cases of anencephalia, 384 cases of spina bifida and 80 cases of encephalocele, 425 cases of congenital hydrocephalus, 261 cases of omphalocele and 258 cases of gastroschisis, 225 cases of congenital defects of oesophagus and 279 anorectal malformations; 232 cases of diaphragmatic hernia, 411 cases of agenesis/hypoplasia of kidneys and 438 cystic kidneys; and 1,412 cases of Down's Syndrome. The success rate of secondary prevention was high for the defects of neural tube (55-96%), for the defects of abdominal wall (55-78%) and for the Down's Syndrome (59%). As concerns the defects treatable by surgery, the survival rate in the first year of life of the born children was lowest in the case of congenital hydrocephalus (63%) and in the case of neural tube defects (75-84%). However, in the case of GIT congenital defects (the defects of abdominal wall, diaphragmatic hernia, congenital defects of oesophagus and anorectal malformations), 82 to 91% of the born children survived. CONCLUSION: Immaturity and concurrent incidence of additional birth defects prolong the period of hospitalisation, UPV and total parenteral nutrition in the case of the affected neonates. This deteriorates the prognosis of the children born with birth defects. Possible reduction of the incidence of these cases could be achieved through improvement in prenatal diagnostics, specifically through more timely identification of mainly the serious types of defects with the presence of associated congenital defects. Another possibility to improve the survival rate of children is the development of surgical procedures and specialised neonatal and infant intensive care. PMID- 15748027 TI - [Week of pregnancy in which birth defects were prenatally diagnosed in the Czech Republic in 1996-2002]. AB - OBJECTIVE: To evaluate the week of pregnancy in which birth defects were prenatally identified and the week of pregnancy in which pregnancy was terminated due to these defects in the Czech Republic in the period from 1996 to 2002. As regards data on Down's Syndrome, comparison to the data from other countries. DESIGN: A retrospective study, which identifies the week of pregnancy when congenital defects are identified through invasive as well as non-invasive prenatal diagnostics and the week of pregnancy when the pregnancy is terminated because of these defects. METHODS: The data were obtained from individual medical genetics facilities. The data are stated for all defects prenatally diagnosed as well as for the defects, which are most frequently prenatally diagnosed - from among neural tube defects, anencephaly and spina bifida were covered; from among abdominal fissures, omphalocele and gastroschisis were covered. Out of inborn chromosomal aberrations, the most frequently diagnosed chromosomal aberration - Down's Syndrome - was covered. RESULTS: The average time of prenatal diagnostics of the birth defects was 19.48 week and the average time of termination of pregnancy due to these defects was 20.11 week. Anencephaly was diagnosed after 17.16 weeks and terminated after 17.84 weeks of the pregnancy on the average. Prenatal diagnostics of spina bifida was carried out after 19.25 weeks and terminated after 19.70 weeks. Omphalocele was identified after 17.55 weeks and terminated after 17.99 weeks. The second type of defect of abdominal wall - gastroschisis was identified after 17.93 weeks and terminated after 18.65 weeks. On the average, Down's Syndrome was diagnosed after 19.81 weeks and terminated after 20.43 weeks. It can be said about all the defects that during the monitored years there was a slight reduction of the period after which the defect was identified as well as of the period after which pregnancy was terminated. CONCLUSION: For all the presented selected birth defects as well as for the defects as a whole, it can be said that during the monitored years there was a slight reduction of the average period after which the defects were identified and as well as of the average period after which pregnancy was terminated. PMID- 15748025 TI - [Condition of the children born after assisted reproduction in Czech Republic]. AB - OBJECTIVE: A retrospective study of data, with an analysis of bio-social factors and the incidence of birth defects in the children born out of pregnancy resulting from assisted reproduction in the Czech Republic in the period from 1995 to 2002. Also, the objective was to analyse these factors in comparison with a group of children born during the same period after spontaneous conception. DATA SOURCE AND METHODS: The data were obtained from the National Health Registers, the administration of which has been entrusted to the Institute of Health Information and Statistics of the Czech Republic (IHIS CR). Specifically, the National Register of Birth Defects and the National Register of Neonates were used. Groups of children born after spontaneous conception and after assisted reproduction were analysed. We analysed the frequency of pregnancy, the gestation week at birth and birth weight, the incidence of immaturity at birth, hypotrophy, hypertrophy, the incidence of birth defects in prenatally and postnatally diagnosed cases and the mortality rate during the 1st year of life. RESULTS: In the period 1995-2002, a total of 731,617 children were born in the Czech Republic. During the same period, a total of 14,243 children were born after assisted reproduction. A statistically significant higher (p<0.0001) proportion of the children born out of multiple pregnancies (40% in the assisted reproduction group as compared with 1.7% in the spontaneous conception group) was identified. The overall mortality rates (early neonatal mortality and infant mortality) are significantly influenced by the number of children within a pregnancy, and as a consequence of the unfavourable distribution in the assisted reproduction group (statistically significant higher representation of children from multiple pregnancies), the rates are higher in the assisted reproduction group. CONCLUSION: The frequencies of children born with a birth defect in the assisted reproduction group and in the group of other born children were not statistically significant. The differences in mortality rates, birth weights and the gestation week are caused by the unfavourable demographic distribution in the assisted reproduction group. PMID- 15748028 TI - [Prognosis of the reproduction health care in the Czech Republic until 2010. I. Perinatal care]. AB - OBJECTIVE: A prognosis of the development of perinatal care in the Czech Republic until 2010 DESIGN: An epidemiological study to be used as a basis for the prognosis. SETTING: Mother and Child Care Institute, Prague. METHODS: The primary source data on the indicators of the level of perinatal care as well as on some circumstances under which this care is being developed and on which their prognosis is based were drawn from the information obtained from their annual nationwide analysis. RESULTS: The prognosis of the development of perinatal care until 2010 builds on the previous prognosis from the period 1980-2000, whose results, when analysed retrospectively, proved a significant dependence of professional development on political, demographic, social and economic circumstances. Increase of the level of perinatal care, evaluated according to internationally recognised criteria, resulted in decreasing the maternal, perinatal and infant mortality rates to levels reported by a few countries in the world. The fact that these levels have been stagnating since 2000 is a result of two conflicting processes. On one hand, there is the improvement in the standard of care; on the other hand, there is deterioration in some of the above-mentioned circumstances. The fact that the reserves for further decrease in professional care have been used up and, conversely, the risk that some of the above-mentioned circumstances will deteriorate have raised concerns that some of the indicators of this care will deteriorate as shown by the extrapolation of the trend in these indicators prevalent in the last years carried out for the period until 2010. Two prognoses with differently serious impacts were formulated on the basis of this extrapolation. CONCLUSION: The development of perinatal care is dependent on the circumstances under which it is being developed. The fact that the reserves in development and organisation of this specific field have been used up, which resulted in decreasing the maternal, perinatal and infant mortality rates to levels, which are among the lowest ones in the world, and, conversely, the deterioration in some of the above-mentioned circumstances have raised concerns that the internationally recognised indicators of this care will deteriorate. PMID- 15748030 TI - [Further development of neonates with severe pneumopathy]. AB - OBJECTIVE: To assess the risk and the degree of possible unfavourable development in neonates with bronchopulmonal dysplasia and with congenital diaphragmatic hernia. DESIGN: Longitudinal monitoring of the mental, motoric and neurological development until the age of 2 years. Comparison of the findings with the result of polygraphic examination carried out at the expected time of birth. METHODS: The following children were monitored: a) 38 children with bronchopulmonal dysplasia born between the 24th and 33rd post-conception weeks; b) 18 children with congenital diaphragmatic hernia, of whom 3 children were born between the 34th and 36th post-conception weeks, while the remaining 15 children were born on the expected date. All children underwent polygraphic examinations (EEG - 8 channels, respiration, EKG, EOG and EMG) between the 38th and 42nd post conception weeks. During the 1st and the 2nd years, their psychomotoric development and neurological status were repeatedly evaluated according to Bayley and Vlach, respectively. RESULTS: Up until the 2nd year of life, approximately 2/3 of the children in both monitored groups were developing favourably or satisfactorily. However, the development of the remaining 1/3 was not favourable: The children were mentally or motorically retarded and/or cerebral palsy developed in them. The polygraphic examination in the neonatal period predicted the favourable or unfavourable development of the monitored children with high probability. CONCLUSION: The study showed that in accordance with the data stated in the literature, the development of neural functions is seriously affected in a relatively high percentage of neonates with severe pneumopathy. Neonatal polygraphy can contribute to timely determination of the prognosis for these children. Children with serious neonatal pneumopathies need to be provided with dispensary care on a long-term basis, and professional consultancy and intervention programmes need to be ensured for them. PMID- 15748029 TI - [Prognosis of the reproduction health care in the Czech Republic until 2010: II. Planned parenthood]. AB - OBJECTIVE: To prepare a prognosis of development of contraception, artificial termination of pregnancy and asisted reproductive technologies until 2010, based on the trend in their indicators in the previous years. DESIGN: A retrospective epidemiological study to be used as a basis for the prognosis SETTING: Mother and Child Care Institute, Prague - Podoli METHODS: For outlining the probable trend in the use of various types of contraceptives (IUD) as well as for artificial termination of pregnancy, evaluated according to their internationally recognised indicators, the extrapolation of the trends in these criteria prevalent in the period 1995-2002 was used. RESULTS: The prognosis of increase in the number of female users of hormonal contraception at the expense of IUD as well as the decrease in the number of women solving their problem with unwanted pregnancy by artificial termination of pregnancy (ATP), based on the said extrapolation of their indicators with a prospect until 2010, is probably overly optimistic. Due to probable changes in the political, social and economic circumstances as well as in the women's attitudes, it is expected that this trend will slow down. This is why it is expected that the number of female users of hormonal contraception will increase merely to 500 and that the women using IUD will decrease to 50 per 1,000 fertile women. Also, as concerns the number of ATPs, it is expected that it will decrease merely just below 10 for every 1,000 fertile women and to 20 for every 100 births. Even if the trends of all these indicators slow down, it can be expected that they will reach the average levels reported in the countries of Western Europe. As for the trend in asisted reproductive technologies (ART), whose share in the total number of births will rise, the main emphasis will be put on consistent nationwide registration, amendments to legislation and research mainly in the area of psycho-neurological development of these children, as recommended by the WHO. CONCLUSION: The expected rate of growth of the number of female users of effective contraception and the expected rate of decline of the number of ATPs, corresponding to the trend of the recent years, will probably decrease by 2010 but the average levels reported by the countries of Western Europe will still be reached. According to the recommendation of the WHO, the trend in some forms of ART will require research in the field of psycho neurological development of the children from these pregnancies until they reach adulthood as well as some amendments to legislation. PMID- 15748031 TI - [The incidence of serious complications in neonates with birth weight < 1,500 g in the Czech Republic (in the period 2000-2002)]. AB - OBJECTIVE: To analyse the incidence of serious complications in neonates with birth weight lower than 1,500 g in the Czech Republic in the years 2000-2002. DESIGN: A retrospective epidemiological study. METHODS: Frequency analysis of the data from the National Register of Neonates, administrated by the Institute of Health Information and Statistics of the Czech Republic. RESULTS: The incidence of PIVH of level III-IV in neonates with weight of up to 1,500 g in the Czech Republic in the years 2000-2002 was 6.8-10.3%; the incidence of cPVL reached 6.4 7.6%; the incidence of ROP of level III and of a higher level was 6.6-7.9% and the incidence of NEC was between 4.3 and 5.7% of all live births. Among the surviving neonates, the percentages of those who did not have any of the PIVH, ROP, HIE or meningitis diagnoses in the reference period were as follows (according to their birth weights): 500-749g 50-63.8%; 750-999 g 69.7-74.8%; 1,000-1,499 g 86.8-91.4%; 1,500-1,999g 98-98.2%; 2,000-2,499 g 99.4-99.5%; and in the group with weight above 2,500g, the percentage was 99.9%. CONCLUSION: The incidence of serious complications in neonates with birth weight of up to 1,500 g in the Czech Republic in the years 2000-2002 was comparable with the data stated in the literature, except for cPVL, the incidence of which was higher in the Czech Republic. The proportion of neonates, who were released and did not have any of the complications, which pose a risk to normal neurosensory development, does not virtually change for birth weights higher than 1,500 g and reaches 98 and more percent. However, among the surviving neonates with birth weight of up to 750, their proportion is mere 50-64%. PMID- 15748032 TI - [New Apgar score? A multi-centre study dealing with the evaluation of the neonate NEOMOD scoring system for the first day of life]. AB - OBJECTIVE: To compare the predictive accuracy of the NEOMOD score and the Apgar score as concerns prediction of mortality and affection of the central nervous system (CNS) in neonates. DESIGN: Prospective multi-centre clinical study. SETTING: Mother and Child Care Institute, Prague; Thomayer's Faculty Hospital with a Policlinic, Prague. METHODS: All neonates with a gestation age > or =35 weeks of pregnancy, hospitalised in the intensive care units of the cooperating hospitals in the period 1998-2002, were included in the study. For all the neonates, the NEOMOD score was established in the first 24 hours and the Apgar score was established in the 1st, 5th and 10th minutes of their lives. The aim was to establish the predictive accuracy of the scoring systems for mortality and affection of CNS in the first 28 days of life. RESULTS: 620 patients participated in the study; 16 patients (1.90%) died by the 28h day. The predictive accuracy for mortality expressed as AUS - the area under receiver operating characteristic (ROC) curve reached the highest level for NEOMOD AUC = 0.96. AUC for the Apgar score was 0.84 in the 1st minute; it was 0.84 in the 5th minute and it was 0.88 in the 10th minute. The predictive accuracy for affection of CNS was AUC = 0.76 in the case of NEOMOD; in the case of Apgar, AUC was 0.71, 0.74 and 0.77 in the 1st, 5th and 10th minute, respectively. CONCLUSION: The NEOMOD system proved a high accuracy in the prediction of neonatal mortality in a group of patients with a gestation age > or =35 weeks of pregnancy. The predictive accuracy of NEOMOD was higher than that of the Apgar score. The predictive accuracy of both the NEOMOD and Apgar scores for affection of CNS in the neonatal period was low. PMID- 15748033 TI - [Influence of oral colonization with probiotic E. coli strain after birth on frequency of recurrent infections, allergy and development of some immunologic parameters. Long-term studies]. AB - OBJECTIVE: prevention of repeated infections and allergies in children of allergic mothers by oral colonization with probiotic E. coli strain.The development of some immunologic parameters. Long - term studies. DESIGN: Original contribution SETTING: Mother and Child Care Institute of Prague. METHODS AND RESULTS: The results of our long-term studies confirmed that orally administered probiotic E. coli strain after birth rapidly colonized the gastrontestinal tract of the newborn and remained dominant for many weeks. The long-term presence of the strain in the intestine stimulated local and serum antibody response. Early induction of secretory IgA production is important particularly in formula-fed infants. The long-term presence of the E. coli strain in the intestine decreased the numer of pathogens colonizing intestinal and other mucous membranes , the frequency of infections and reduced need for antibiotics in premature and high risk infants. Ten years later, there was still a lower frequency of repeated infections (23%) in comparison with control children (58%). Colonization with probiotic E. coli strain in infants treated in protected (pathogen-free) environment represented effective prevention of nosocomial infections In the colonized group infections occured in 16% of infants and 130 isolates and 7 genera of pathogens were demonstrated. In the group treated in conventional environment 40% of infants had nosocomial infections, 238 isolates and 10 genera of pathogens were proved. The hospitalization period was shorter in the first group (26 versus 34 days). Intentional colonization with probiotic E. coli after birth reduced incidence of allergies after 10 and 20 years (being 12% and 16% in the colonized groups and 33 and 32% in controls). In the present long - term study (evaluated after the first year) colonization with vaccine COLINFANT after birth influenced the levels of some cytokines ( IL-4, IFN-gama,TGF-beta) and also clinical manifestation of allergy (there were no signs of allergy in colonized infants of allergic mothers, but 25% of infants of control allergic mothers had clinical manifestations of allergies). CONCLUSIONS: By replacement of the natural but incidental ( event. pathogenic ) colonization of the intestine by a targeted orally administered E. coli strain after birth we may have come upon the possibility of how to prevent nosomial infections particularly in formula-fed and high-risk infants and prevent occurence of allergies in infants of allergic mothers. PMID- 15748034 TI - [Prediction of type 1 diabetes in the neonates of diabetic mothers or fathers]. AB - OBJECTIVE: Verification of the possibility to predict diabetes in the neonates of mothers and fathers with Type 1 diabetes. DESIGN: Prospective clinical study. SETTING: Mother and Child Care Institute, Prague. Paediatric Clinic of the 2nd Faculty of Medicine at the Charles University, Prague. METHODS: In 31 neonates of mothers and fathers with Type 1 diabetes, the long-term and short-term risk of the occurrence of Type 1 diabetes was established. The genotypification of HLA, DQB1, HLA DQA1 and DRB1 *04 was carried out by using the PCR method to establish the long-term risk and according to the result of the examination, the examined child was included into one of the five categories of genetic risk. In all the monitored persons, the levels of antibodies against GAD65, IA-2 and IRA insulin were repeatedly identified by means of the methods, which are the markers of autoimmune insulitis and show the short-term risk of the occurrence of diabetes. The function-related examination of secretion of beta cells was carried out by using the intravenous glucose tolerance test (i.v. GTT) in children with significant titres of one or more antibodies. RESULTS: A very high risk of the occurrence of Type 1 diabetes was identified in 1 child with the genotype DQA1*03 DQB1*0201/DQA1*05-DQB1*0302 (3.23%); an increased risk was identified in 12 children (38.71%); a medium risk was identified in 11 children (35.48%); a relatively low risk was identified in 3 child (9,68%) and a low risk was identified in 4 children (12,90%). In 4 children (12.9%), a strongly protective alelle DQB1*0602 was found. In 4 children, positivity for one of the antibodies was identified. In 1 child of a diabetic father with an increased genetic risk, there was a decrease in the titre of antibodies in the case of repeated check and function-related examination of the insulin secretion (FPIR) will be carried out. In another child, disappearance of antibodies was identified when samples were taken for verification; function-related examination of insulin secretion produced normal results, and the child has remained without clinical manifestation of diabetes. In a third child, the positivity of the antibodies from the umbilical blood was only temporary and was probably caused by a passive transfer from the mother; now, when repeated checks were made, the antibodies have remained negative. In a fourth child, the parents refused further examinations after antibody positivity was observed; the child has been without clinical manifestation of diabetes up until now. CONCLUSION: This scheme for predicting diabetes by means of immunogenetic and immunological examination of risk individuals is a rational measure aimed at timely identification of autoimmune insulitis, which precedes the occurrence of Type 1 diabetes, and it should become a standard part of diabetological care. PMID- 15748035 TI - [The first experience with linezolide in treatment of nosocomial septic condition in premature neonates]. AB - OBJECTIVE: Nosocomial infections caused by the coagulase-negative staphylococci are among the most frequent complications in an intensive care unit for neonates. The authors describe three cases of successful treatment of this complication in which the new antibiotic Linezolide was used. DESIGN: Analysis of three clinical cases. SETTING: Mother and Child Care Institute, Prague. METHODS: Description of cases of nosocomial sepsis in 3 immature neonates where the usual treatment of multiresistant strains of coagulase-negative staphylococci, based on the use of Vankomycin, was not successful. CONCLUSION: Administration of Linezolide proved to be very effective in all of the described cases. In spite of the high effectiveness and relative safety of this preparation, its precise role in the treatment of infection-related complications caused by coagulase-negative staphylococci has not yet been established. For the time being, we recommend administering this preparation only in strictly indicated cases after the usual therapy fails. PMID- 15748037 TI - [Effect of early nutrition on growth parameters and psychomotor development of children of very low birth weight]. AB - OBJECTIVE: The aim of the study was to evaluate influence of early nutrition on growth parameters and psychomotor development of children with very low birth weight (VLBW). DESIGN: A prospective clinical study. SETTING: Institute for Care of Mother and Child, Prague. METHODS: Thirty nine children of birth weight 1,000 1,499 were followed up to one year of their corrected age in a prospective study. The group was divided in two groups according to type of nutrition: 17 children (group A) were fed with milk of own mother - "preterm milk", 22 children (group B) were orally fed with mature milk from the Bank of mother milk - "term milk", which was fortified with BMF preparation (Nutricia, Netherlands). Both groups were comparable in basic anthropometric parameters (weight, lenght, circumference of head and thotax) and in psychosocial characteristics of their mothers. Growths parameters were monitored in weekly intervals for approximatelly eight weeks. In the period between 11th and 15th month of corrected age, the children were evaluated by a clinical psychologist on a blind basis in mental a motor development by using Bayley Scales of Infant Development (BSID-II). Statistical analysis was performed by chi-square test and t-test. RESULTS: No statistically significant differences between the two groups in evaluating the growth parameters were observed. The psychological examination demonstrated statistically significant differences in the motor development. The psychomotor developmental index (PVI) proved to be 84.4 +/- 14.6 in the group A and 94.3 +/- 12.5 in the group B (t-test = 2.28, p<0.05). There was not any statistically significent difference in metal development between the two groups. The mean mentel developmental index (MVI) was 98.2 +/- 10.2 in the A group and 101.0 +/- 13.3 in the group B. CONCLUSION: Result of the study indicate favorable effect of fortification of breast milk in VLBW newborns, especially in view of the observed favorable influence of fortfication on motor development of the children. PMID- 15748036 TI - [Umbilical stump care in full-term neonates]. AB - OBJECTIVE: The objective of the study was to evaluate the umbilical stump care in the Perinatological Centres in the Czech Republic and to propose a uniform therapeutic procedure in relation to the issue of early release of full-term neonates. DESIGN: An epidemiological data analysis. SETTING: Mother and Child Care Institute, Prague - Podoli. METHODS: The representatives of all the Perinatological Centres (PCs) in the Czech Republic (12 centres) were contacted over the telephone with the aim of evaluating the umbilical stump care in full term neonates in relation to the period of hospitalisation. The telephonic questionnaire evaluated the following aspects: the treatment method used in the first days after birth; the time of surgical removal and/or leaving the umbilical stump; the technique of surgical access; the treatment after ablation of umbilical stump and the period of hospitalisation. RESULTS: In the first days, antiseptics are used in all the Perinatological Centres of the Czech Republic. All PCs carry out surgical removal of umbilical stump (in 7 centres it is cut off and in 5 centres it is removed through abscission). In 11 centres, the surgical removal takes place 36-48 hours after birth; in 1 centre it takes place 60-72 hours after birth. After removal, antibiotics are applied locally in 2 centres and antiseptics are applied locally in 10 centres. The period after which neonates were released was > or = 72 hours in 8 centres and > or = 96 hours in 4 centres in the case of full-term neonates after spontaneous birth. CONCLUSION: The uniform practice of the Perinatological Centres in the Czech Republic is sterile surgical removal of umbilical stump. However, with regard to early release and after-care, the current trend is to prefer a non-surgical method with spontaneous mummification and detachment of umbilical cord. PMID- 15748038 TI - [Psychomotor development of children with signs of intrapartum hypoxia and monitored by intrapartum fetal pulse oxymetry]. AB - OBJECTIVE: The following questions were investigated: 1) the psychomotor development in newborns with pathological cardiotocographic recording (CTG), having been born spontaneously due to a normal oxygen saturation monitored by intrapartum pulse oxymetry (IFPO)(group I), 2) possible difference in development of children in group I in comparison with those with pathological CTG without IFPO monitoring delivered by cesarean section (group II), 3) possible differences in children in these two groups with signs of intrapartum hypoxia (groups I and II, respectively) from children delivered spontaneously during physiological delivery without signs of hypoxia (group III). DESIGN: A prospective clinical study SETTING: Institute for the Care of Mother and Child METHODS: There have been 88 children born in the years 2000-2001 from singleton pregnancies in gestational ages > 36 weeks. Three groups of children were established: 1) in the group I (IFPO, n=32) there were children born spontaneously having pathological CTG detected by cardiotocograph HP50A and simultaneously monitored by intrapartum fetal pulse oxymeter NELLCOR Puritan Benett A400 with the NELLCOR FE 14 sensor with normal values of oxygen saturation. The children of group II (CTG-SC, n=25) were born by cesarean section due to acute intrapartum fetal hypoxia (IH) indicated by CTG recording without IFPO. The group III (physiological, n=31) was formed by children from physiological deliveries without signs of IH. All children underwent psychological examination at the average age of 21.8 months by the Bayley Scales of Infant Development (BSID-II). The statistical analysis of data was performed according to the variables using t-test or chi-square test, respectively. RESULTS: Group I (IFPO) did not differ significantly from group II (CTG-SC) in any area of psychological evaluation. The prevention of cesarean section due to IFPO method in the group I therefore prevented a significant increase of psychological morbidity in children of this early age. In spite of that the children of the group I (IFPO) reached the lowest evaluation and differed significantly from the most favorably evaluated group III (physiological) in motor development, behavior, and in a total number of deviations. Group II (CTG-SC) was evaluated significantly worse in comparison with group III (physiological) only in behavior items. On the other hand the occurrence of children of above-average mental development in the groups I and II were the same as in group III. The results indicate complex character of the problem. CONCLUSION: The prevention of CS due to IFPO prevented a development of psychological morbidity in children. The increased occurrence of deviations in psychomotor development of children with signs of acute IH (groups I and II) in comparison with children of group III (physiological) will be analyzed in detail in relation to further data. PMID- 15748039 TI - [Are there any new procedures for treating necrotising enterocolitis in neonates with very low birth weight?]. AB - OBJECTIVE: To evaluate some perinatal and neonatal risk factors of occurrence of necrotising enterocolitis (NEC) and the current possibilities of treatment. DESIGN: A retrospective data analysis. SETTING: Mother and Child Care Institute, Prague. METHODS: The clinical course in 28 neonates treated for NEC on the worksite of the authors in the period 1999-2003. Only neonates with NEC in the second and third Bell's stages were included in the group. RESULTS: From a group of 14,275 neonates born during the reference period, NEC was diagnosed in 28 neonates. In all cases, they were neonates with very low birth weight (VLBW). The average gestation age was 26.6 weeks (the range was 23-35; the median was 25.5); the average birth weight was 822 grams (the range was 340-1,490g; the median was 735). Incidence was between 1.08 and 2.37/1,000 live births (the average was 1.94). On the average, NEC occurred on the 17th day after birth (the range was 5 59; the median was 14). 14 neonates (50%) in the acute stage of NEC were operated on; 9 children (32%) were only treated conservatively. In eight children at the acute stage, who were operated on, divided ileostomy was created; in four children, a T-drain was used for removal of the intestinal content and an abdominal drain was used for this purpose in one child. Repeated surgery was necessary in five children. Of the conservatively treated ones, five children recovered as a result of conservative treatment and four children died as a result of fast progression of NEC without surgical treatment. All the children treated for patent ductus arteriosus (PDA) with Indometacin died. The overall mortality in the group was 39.3% (11 children died). CONCLUSION: Prematurity seems to be the common denominator of the occurrence of NEC. Treatment of NEC in highly immature neonates with birth weight below 1,500g requires comprehensive cooperation of a neonatologist and a children's surgeon. Resection of a necrotic intestine with creation of a stomy remains a standard procedure, which is followed in unstable neonates with a localised form of the illness. Prevention of premature birth, antenatal steroids, frequent feeding (trophic feeding) and the corresponding correction of water management seem to be a reasonable strategy to reduce the incidence of NEC. The resulting incidence, mortality, the time of the occurrence of NEC and the therapeutic procedures are within the limits stated in the international literature. PMID- 15748040 TI - [Influence of antenatal administration of corticoids on heart activity--case report]. AB - OBJECTIVE: to draw attention to possible influencing of cardiotocographic recording of antenatal administration of corticoids, on the use of Doppler ultrasonographic parameters in the diagnostics of hypoxia-endangered fetus. DESIGN: Case report. SETTING: Mother and Child Care Institute of Prague. METHOD: Case report. CONCLUSION: Antenatal administration of corticoids is presently an inherent part of therapeutic management of premature delivery. This administration significantly diminishes morbidity and mortality of immature newborns. Nevertheless, their administration results in changing intrauterine behavior of the fetus and his/her heart activity. These changes may be mistakenly diagnosed as signs of evolving hypoxia. For the evaluation of acute condition of the fetus and obtaining help in decision making how the further management of delivery should be done, Doppler ultrasonographic examination of maternal, fetus maternal and fetal circulation may prove useful. PMID- 15748041 TI - Alcohol and illicit drug dependence. AB - OBJECTIVES: This article estimates the prevalence of alcohol and illicit drug dependence among Canadians aged 15 or older Comorbidity with depression is examined. DATA SOURCES: The data are from the 2002 Canadian Community Health Survey: Mental Health and Well-being and the National Population Health Survey. ANALYTICAL TECHNIQUES: Cross-tabulations were used to estimate the prevalence of alcohol and illicit drug dependence by selected characteristics. Multiple logistic regression models were used to determine if associations persisted after controlling for potentially confounding factors, and to test temporal relationships between frequent heavy drinking and depression. MAIN RESULTS: In 2002, an estimated 641,000 people (2.6% of the household population aged 15 or older) were dependent on alcohol, and 194,000 (0.8%), on illicit drugs. These people had elevated levels of depression compared with the general population. Heavy drinking more than once a week was a risk factor for a new episode of depression, and depression was a risk factor for new cases of frequent heavy drinking. PMID- 15748042 TI - Bipolar I disorder, social support and work. AB - OBJECTIVES: This article reports the estimated lifetime prevalence of bipolar I disorder in the household population and describes characteristics of people of working age (25 to 64) affected by this disorder. The relationship between social support and employment status is examined in people with the disorder. DATA SOURCE: Data are from the 2002 Canadian Community Health Survey: Mental Health and Well-being. ANALYTICAL TECHNIQUES: Weighted frequencies and cross-tabulations were used to estimate the prevalence of bipolar I disorder. Multiple logistic regression modeling was used to examine four dimensions of social support in relation to having a job, in people with bipolar I disorder. MAIN RESULTS: An estimated 444,000 (2.6%) people aged 25 to 64 had lifetime bipolar I disorder. Alcohol dependence, asthma, migraine, obesity and panic disorder were far more prevalent among these people, compared with the general population. People with bipolar I disorder who reported readily accessible tangible support had higher odds of being employed, compared with those with less available tangible support. PMID- 15748043 TI - Panic disorder and coping. AB - OBJECTIVES: This article presents prevalence estimates of panic disorder in the household population aged 15 or older Associations between panic disorder and measures of physical and mental health, work status and coping behaviour are examined. DATA SOURCE: Data are from the 2002 Canadian Community Health Survey: Mental Health and Well-being. ANALYTICAL TECHNIQUES: 2002 prevalence rates are presented for people with a history of panic disorder. Characteristics associated with current and past panic disorder are examined. Multiple logistic regression models are used to examine work status and coping behaviour, and chronic physical and other mental health problems. MAIN RESULTS: In 2002, an estimated 1.5% of the population had current panic disorder, and 2.1%, a past history. Average age of onset was 25. People with current panic disorder were less likely to work and more likely to be permanently unable to work, compared with those who had never had the condition. Negative coping behaviours, including alcohol or drug use and smoking, were more common among those with panic disorder. PMID- 15748045 TI - Some of the age-old communicable diseases remain a significant public health problem. PMID- 15748044 TI - Social anxiety disorder--beyond shyness. AB - OBJECTIVES: This article presents prevalence estimates of social anxiety disorder (social phobia) among the Canadian household population aged 15 or older. The relationship between this mental disorder and others is examined. Selected aspects of functional impairment are compared for people with current, past, and no history of the condition. DATA SOURCE: Data are from the 2002 Canadian Community Health Survey: Mental Health and Well-being. ANALYTICAL TECHNIQUES: Cross-tabulations were used to estimate the prevalence of social anxiety disorder, to determine socio-economic factors associated with prevalence, and to examine relationships with other mental disorders. Associations between social anxiety disorder and selected impairment variables were examined using multivariate analysis that controlled for socio-economic factors and other aspects of mental and physical health. MAIN RESULTS: In 2002, 750,000 Canadians aged 15 or older (3%) had social anxiety disorder. These people had a higher risk of having major depressive disorder, panic disorder and substance dependency than the general population. Social anxiety disorder was associated with higher rates of disability, negative perceptions of physical and mental health, and dissatisfaction with life. PMID- 15748046 TI - The special issue for tropical disease research: a tool to disseminate research results for more effective disease control. PMID- 15748047 TI - Antimalarial drug resistance in the Eastern Mediterranean Region. AB - Studies done between 1997 and 2003 are reviewed to give an overall picture of antimalarial drug resistance in the Eastern Mediterranean Region of the World Health Organization (WHO). The situation in 8 countries where resistance has been reported is detailed. It has been difficult to abandon chloroquine as first-line treatment even though resistance to it is widespread. Resistance to sulfadoxine pyrimethamine has also been detected. The spread of resistance could be slowed down by the adoption of effective national policies and control programmes. Coordination between counties and with other WHO regions, as opposed to working in isolation, could further reduce the spread of resistance. PMID- 15748048 TI - Controlling tuberculosis: we can't do it if we don't find the cases. AB - The current international strategy for the control of tuberculosis emphasizes two different targets: a high success rate for the treatment of tuberculosis cases with an emphasis on positive sputum smear cases and a high case detection rate. Recent analyses indicate that while progress is being made towards successful treatment, targets for case-finding are not being achieved. PMID- 15748049 TI - Recent advances in research and control of malaria, leishmaniasis, trypanosomiasis and schistosomiasis. AB - In the Eastern Mediterranean Region of the World Health Organization (WHO), malaria, schistosomiasis, leishmaniasis and trypanosomiasis are the parasitic diseases of major importance. Our review focuses on recent advances in the control and treatment of these diseases with particular reference to diagnosis, chemotherapy, vaccines, vector and environmental control. The Roll Back Malaria Programme, for example, emphasizes the use of insecticide treated bednets in Africa and targets a 30-fold increase in treated bednet use by 2007. Increasing risk factors for leishmaniasis include urbanization, extended agricultural projects and civil unrest and the increase in patients with Leishmania infantum and HIV co-infection in the Region may signal a new threat. In the past 20 years, human African trypanosomiasis has resurged in sub-Saharan Africa; within the Region it has become more common in the southern Sudan where anthroponotic and zoonotic sub-species infections overlap. Schistosomiasis in the Region is caused by either Schistosoma haematobium or S. mansoni and large-scale control efforts include providing regular treatment to at-risk groups and supporting drug delivery through schools. PMID- 15748051 TI - Estimation of malaria transmission in high-risk provinces of Morocco. AB - The malaria transmission level of Plasmodium vivax was monitored in four high risk provinces in Morocco. Intensive mosquito collection by light traps and manual catches resulted in the capture of four species: Anopheles labranchiae, An. sergenti, An. cinereus, and An. claviger. All An. sergenti and An. labranchiae females collected were tested for the presence of two phenotypes of P. vivax (PVK210 and PVK247) antigen by enzyme-linked immunosorbent assay (ELISA). No P. vivax antigen was detected in 1347 mosquitoes analysed. A parallel parasitological investigation was conducted. Of 2665 slides examined from a population of 4343 people for detection of P. vivax, no slide was positive. The results confirm the break in malaria transmission in residual foci. The use of ELISA is recommended in future epidemiological studies of human malaria. PMID- 15748050 TI - Lymphatic filariasis in the Eastern Mediterranean Region: current status and prospects for elimination. AB - Lymphatic filariasis (LF) represents a major public health problem in tropical and subtropical regions of the world. The disease is endemic or suspected in several countries of the Eastern Mediterranean Region. Recent advances in diagnosis and therapy led the World Health Assembly to pass a resolution in 1997 calling for "the elimination of lymphatic filariasis as a public health problem." The elimination strategy is based on rounds of mass drug administration of an annual single-dose of combined drug regimens for 5-6 consecutive years. Subsequent steps included formation of a Regional Programme Review Group to orient national LF control programmes towards the concept of elimination, provide advice, review each national plan of action and review annual reports. To date, Egypt and the Republic of Yemen have active national LF elimination programmes, however, elimination activities in the Republic of Yemen are still restricted to certain identified endemic regions. Other countries in the Region are on their way to verifying the situation and if LF is proved to be endemic, will start mapping endemic localities. This review sheds light on the status of LF elimination activities in the Region and highlights some of the major accomplishments. PMID- 15748052 TI - GIS-based prediction of malaria risk in Egypt. AB - Environmental variables in a malaria geographic information system (GIS) database were analysed to discriminate between governorates at high and low risk of malaria. Only Fayoum governorate was categorized as a high risk area for malaria during the last 2 decades. Discriminant models correctly classified 96.3% of the risk categories and indicated that the most important predictor of risk is hydrogeology. Further GIS spatial analysis indicated that the high malaria risk in Fayoum is associated with a unique environmental envelope of biotic (presence of both efficient malaria vectors) and abiotic (hydrogeology and soil) variables. Recommendations for surveillance and control are discussed. PMID- 15748053 TI - Stratification of Khartoum urban area by the risk of malaria transmission. AB - A cross-sectional community-based study was carried out in Khartoum urban area aimed at stratifying the area by risk of malaria transmission. Two thousand households (8092 individuals) were surveyed during the cold, dry and rainy seasons of 2002. Households with screened windows, using bednets or both were 5.0% (95% CI: 4.1-6.0), 10.9% (95% CI: 9.5-12.3) and 1.8% (95% CI: 1.2-2.5) respectively. Access to health services was high. Only 3.8% (95% CI: 2.9-4.7) of households kept antimalarial drugs at home. The parasite rate, spleen rate and fever rate were very low (0.21%, 0.17% and 0.97% respectively). Overall, people in Greater Bahry suffer a greater burden of malaria than those in Greater Khartoum or Greater Omdurman, with a seasonal pattern of transmission. PMID- 15748055 TI - Determinants of acquiring malaria among displaced people in Khartoum state, Sudan. AB - A cross-sectional descriptive study was carried out among displaced people in Khartoum state to determine risk factors associated with malaria. Data were collected from 856 households about sociodemographic characteristics, history of malaria, and knowledge, attitudes and treatment-seeking behaviour. Overall, 68.2% reported a malaria attack among household members in the previous year. Risk of malaria attack was significantly associated with tribe, language, education, water supply and food expenditure. The highest rates of attack were among local language speakers (85.5%) and illiterate residents (70.4%). Half the respondents (50.2%) delayed seeking treatment for malaria. Knowledge, attitudes and practices had no association with malaria attacks, except for a 4.7-fold increased risk of malaria when obtaining water from carts rather than wells. PMID- 15748056 TI - Determinants of malaria mortality among displaced people in Khartoum state, Sudan. AB - To find the determinants of malaria mortality among displaced people, across sectional descriptive study using verbal autopsy was carried out in 2 camps in Khartoum state, Sudan. The heads of 856 households were interviewed about household characteristics, and malaria knowledge, attitudes and behaviour. They reported 81 malaria deaths during the previous year, 3.5% of all household members; 70 (86.4%) were children. Fever, diarrhoea, vomiting and headache were the most prevalent symptoms before death. Having a malaria death in the household was significantly associated with poor knowledge about malaria and, surprisingly, with better education. Poor treatment-seeking behaviour and poor attitudes towards malaria were not associated with higher mortality. However, mortality was significantly higher among households obtaining water by cart than from a well. PMID- 15748054 TI - Plasmodium falciparum infection during pregnancy in an unstable transmission area in eastern Sudan. AB - A 1-year prospective community-based study of malaria during pregnancy was conducted in an area of seasonal and unstable malaria transmission in eastern Sudan. At a village antenatal clinic, 89 non-pregnant controls and 86 pregnant women were enrolled and followed every 2 weeks until 6 weeks after delivery. The incidence of Plasmodium falciparum infection was significantly higher among pregnant than control women (17.4% versus 5.6%) with no difference between primigravidae and multigravidae (22.2% versus 15.2%). There was no significant difference in the mean haemoglobin concentration between infected and uninfected mothers (9.1 +/- 1.3 versus 9.5 +/- 0.6 g/dL) but the mean birth weight of their babies was significantly lower (2.72 +/- 0.26 versus 2.95 +/- 0.05 kg) despite prompt case management of infected women. PMID- 15748057 TI - Household survey of treatment of malaria in Hajjah, Yemen. AB - The practice of self-medication is widespread in the Republic of Yemen. The objectives of this study were to describe the treatment of malaria in households and to promote rational treatment. We surveyed 201 households with family members suffering from malaria or being treated with antimalarials. Numbers of prescribed and non-prescribed drugs were recorded and treatment rationality assessed. Common patterns of irrational treatment of malaria were observed. Polypharmacy was common, with an average of 3.8 total drugs and 1.3 antimalarials found per encounter. Misused and over use of injectables antimalarials was common. People practised self-medication because of belief, experience, lack of confidence in health services and cost of treatment. Most had no knowledge concerning possible risks of antimalarials. PMID- 15748058 TI - Prescribing rationality and availability of antimalarial drugs in Hajjah, Yemen. AB - This study in 3 districts of Hajjah governorate, Yemen, used WHO core indicators to investigate irrational prescribing of antimalarial drugs in public and private health facilities. Laboratory diagnosis rates were low in public facilities (21.2% of encounters). Informal prescriptions were issued in > 70% of encounters (public and private). Important patient and drug information was missing from many prescriptions. Both public and private facilities had high rates of prescribing multiple drugs (mean 3.0 and 4.0 respectively per encounter, maximum 11), brand-name drugs (32.9% and 64.2%) and injections (17.2% and 33.5%). The total number of antimalarial drugs registered in the country was found to be 98, with 52 different formulations and strengths of chloroquine. Efforts should be made to promote rational prescribing of antimalarials. PMID- 15748059 TI - Study of larvivorous fish for malaria vector control in Somalia, 2002. AB - An intervention study was conducted on the introduction of the larvivorous fish Oreochromis spilurus spilurus as a method of malaria vector control in Kalabeydh village, northern Somalia. This species is resistant to chlorine in water up to a concentration of 1.0 mg/L. Fish were introduced into 25 berkit (reservoirs). After 1 month the number of larvae in each berkit was reduced by between 16.5% and 78.6% (mean 52.8%). Community acceptance and participation was good. The introduction of larvivorous fish is a cheap method of malaria vector control, but its sustainability needs special consideration and education of the community is important, especially to cover the dry season, when most of the berkit dry up. PMID- 15748060 TI - Comparison of three larviciding options for malaria vector control. AB - A field study was carried out over 27 weeks in the south Batinah region of Oman to assess the efficiency and cost-effectiveness of different strategies for vector control of malaria. Three larviciding strategies for Anopheles spp. were applied to intervention areas and compared with a control area, with over 2000 breeding sites monitored for 6 months. The normal method of spraying 1 ppm temephos larvicide fortnightly was found to be less efficient and less cost effective than using 0.5 ppm temephos applied weekly. A third, more environmentally favourable method, to search for vector larvae and treat only those breeding places, was more effective than fortnightly spraying but less effective than the weekly half dose and was the most expensive strategy. PMID- 15748061 TI - Operational use of neem oil as an alternative anopheline larvicide. Part A: Laboratory and field efficacy. AB - We conducted a study to determine the laboratory and field efficacy of neem oil towards anopheline larvae. No difference in LC50 was observed between laboratory and field strains for temephos, chlorpyriphos-methyl/fenitrothion and neem oil. No difference in susceptibility was found after 3 months of application every 2 weeks. Water treated with a single application of traditional larvicides was free of larvae after 4 weeks; neem oil-treated water, however, was free after 2 weeks but not at 4 weeks. Application of chlorpyriphos-methyl/fenitrothion and neem oil every 2 weeks for 7 rounds resulted in dramatic reduction in larval density with no statistically significant differences. An adult survey after larviciding also showed no significant difference. The efficacy of crude neem oil appears to be below that of conventional larvicides. PMID- 15748062 TI - Operational use of neem oil as an alternative anopheline larvicide. Part B: Environmental impact and toxicological potential. AB - This study was conducted to investigate the preliminary environmental and mammalian toxicology of neem oil, temephos and chlorpyriphos-methyl/fenitrothion. Culex pipiens, Daphnia magna and Gambusia affinis were used to study environmental impact. A high level of toxicity was observed, with slight differences between organisms. The emulsifiers individually also displayed toxicity towards the tested organisms. Up to 90 days daily oral crude neem oil treatment (5 g/kg body weight) of laboratory mice did not cause any significant changes in weekly body weight gain, nor in serum liver damage indicators, direct bilirubin or total bilirubin. Blood parameters of treated mice up to 90 days were not statistically different from those of control mice. Neem oil could be used as an environmentally friendly alternative to the traditional chemical anopheline larvicides. PMID- 15748063 TI - Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran. AB - A diagnostic polymerase chain reaction (PCR) assay using species-specific primers and direct sequencing was used to identify members of the Anopheles maculipennis complex in the north-west and central regions of the Islamic Republic of Iran. Specimens were collected from 9 provinces during 2 seasonal activities in 2001 2002, identified morphologically and subjected to PCR assay and direct sequencing. Results showed that only 2 species, An. maculipennis Meigen, and An. sacharovi Faver, were present in the area of study. This was confirmed by the high similarity (99.2%-100%) of their sequences with those available in GenBank. The molecular data and relative distribution of these species in relation to their vectorial capacity and the epidemiology of malaria in the region are discussed. PMID- 15748064 TI - Prevalence of pulmonary tuberculosis in Karachi juvenile jail, Pakistan. AB - Jail inmates may be at increased risk of contracting tuberculosis (TB). We studied 386 detainees (mean age 17.7 years) in Karachi juvenile jail to determine the prevalence of TB and possible risk factors for contracting TB. We found a 3.9% prevalence of TB among the inmates, significantly higher than the estimated 1.1% prevalence in the general population of Pakistan. Positive family history of TB was a significant risk factor for TB. Poor adherence of previously diagnosed patients to anti-TB treatment was found. Our study highlights the vulnerability of inmates to TB owing to the presence of highly infectious cases, along with environmental conditions such as overcrowding and poor ventilation. This study strongly indicates the need for an effective treatment programme in the jails as well in the general community. PMID- 15748066 TI - Predictors of treatment failure among tuberculosis patients under DOTS strategy in Egypt. AB - Treatment failure is a serious problem facing some national tuberculosis (TB) control programmes. Irregularity of treatment is a factor that can lead to treatment failure. A case-control study was carried out in TB centres in Egypt during April 2001-December 2002 aimed at investigating the predictors of treatment failure. We interviewed 119 people with treatment failure and an equal number of cured cases (controls) and their families regarding sociodemographic characteristics, information about TB, information about drugs, treatment compliance, family support and patient-family interaction. Significant risk factors for treatment failure were non-compliance to treatment, deficient health education to the patient, poor patient knowledge regarding the disease and diabetes mellitus as co-morbid condition. PMID- 15748065 TI - Active tuberculosis among Iraqi schoolchildren with positive skin tests and their household contacts. AB - In a prospective cohort study in Iraq, schoolchildren with a positive tuberculin skin test during the nationwide survey in 2000 were followed up in 2002 to determine prevalence of latent tuberculosis (TB) infection and risk factors among household contacts. Of 205 children, 191 remained skin-test positive in 2002. Based on X-ray and clinical examination, 9 children (4.4%) were active TB cases. Among 834 household contacts, there were 144 new TB cases, giving a cumulative incidence of 17.3%. Risk factors for TB among household contacts were: age > or = 15 years; technical/professional job; smoking; low body mass index; diabetes mellitus; steroid therapy; and closeness of contact with the index cases. Based on past history of TB in index children and their contacts, 77.2% of new TB cases were attributable to household contacts. PMID- 15748067 TI - Does routine home visiting improve the return rate and outcome of DOTS patients who delay treatment? AB - Over a period of 6 months the effect of home visits on compliance with directly observed therapy, short course (DOTS), was studied on 480 new smear-positive tuberculosis patients who had delayed collecting their drugs on one occasion. Patients registered at 15 tuberculosis treatment centres in Baghdad, Iraq, were randomized to an intervention group (receiving home visits from trained personnel) or a control group. Home visits were highly effective in improving the return to treatment of patients who were late for treatment (231/240, 96.3%). The intervention group showed a higher treatment success rate (94.2% versus 76.7%), lower default rate (0.8% versus 10.0%) and higher smear conversion rate after the end of treatment (92.9% versus 75.0%) than controls. Home visiting by trained personnel significantly improves patient compliance with DOTS. PMID- 15748068 TI - Impact of community participation on treatment outcomes and compliance of DOTS patients in Iraq. AB - We assessed the effect of community participation on treatment outcomes for tuberculosis patients undergoing directly-observed therapy, short course (DOTS). From February to December 2001 172 newly diagnosed patients in Baghdad were allocated into 2 treatment groups. The intervention group were visited daily at home for the 2-month initial phase by trained members of the Iraqi Women's Federation while the control group attended the local health centre for treatment. Cure rates for patients treated at home were significantly better than controls (83.7% versus 68.6%), so too was compliance (100.0% versus 14.0%). Smear conversion rates were significantly better in intervention cases compared with controls at all stages. Default rates were similar in both groups (11.6% versus 10.5%), as was mortality (1 patient each). PMID- 15748069 TI - Knowledge, attitudes and practices survey among health care workers and tuberculosis patients in Iraq. AB - A cross-sectional study was made of 500 patients and 500 health care workers randomly selected from 250 primary health care centres throughout Iraq to evaluate knowledge, attitudes and practices towards tuberculosis (TB). Using structured questionnaire interviews, the study showed 64.4% of patients had good knowledge, while 54.8% had negative attitudes and practices towards TB. The 2 most important sources of patient information about TB were physicians and television. Of health care workers, 95.5% had good knowledge about TB and this was significantly associated with age and job duration. By contrast, health care workers' practice was poor: only 38.2% handled suspected TB cases correctly. The national TB programme in Iraq has had a good impact on knowledge of TB patients and health care workers. PMID- 15748070 TI - Gender perspectives on knowledge and practices regarding tuberculosis in urban and rural areas in Pakistan. AB - We investigated gender differences in knowledge of and attitude towards tuberculosis (TB) in urban and rural communities in Sindh province, Pakistan. Knowledge of symptoms was generally deficient, particularly in rural females. Regarding TB prevention, 22.4% of rural and 14.4% of urban males said completing treatment was important; only 9.8% of rural and 7.1% of urban females agreed. Doctors were an important source of information in rural areas and 60.9% of rural males said they would only stop treatment on a doctor's advice. In contrast, > 65% of respondents in urban areas said they would stop treatment when symptoms ended. Our study highlights the need to increase population awareness about TB in Sindh. PMID- 15748071 TI - Gender differences in health care utilization and outcome of respiratory tuberculosis in Alexandria. AB - A study of gender differences in health care utilization and outcome of respiratory tuberculosis was carried out in Alexandria, Egypt. A cohort of 334 patients was followed-up for 8 months; 69.2% of cases were males. The pattern of tuberculosis symptoms was similar for both sexes. Women started treatment earlier than men. Women had significantly lower scores in knowledge, beliefs and attitudes about tuberculosis than men. Compliance was unsatisfactory for both sexes. Men tended to be more adherent to drugs and to sputum and X-ray examinations but there were no sex differences in compliance with health education and medical examinations. No significant sex differences in treatment outcome were found: the overall cure rate was 60.5% and treatment failure was 4.8%. Multiple regression analysis showed satisfaction with medical care was the only significant predictor of treatment failure. PMID- 15748072 TI - Gender differences and tuberculosis in the Syrian Arab Republic: patients' attitudes, compliance and outcomes. AB - In a prospective study of gender and TB outcomes, 552 newly diagnosed smear positive patients throughout the Syrian Arab Republic were interviewed at recruitment (from January 2002 to July 2002) and followed until the end of treatment. Delay in diagnosis was significantly longer among males. Significant differences between males and females were noted in relation to the place they usually use to seek care. Gender did not seem to affect knowledge and attitudes of TB patients. Although the women reported more barriers to seeking care, compliance with treatment tended to be higher and the treatment success rate was significantly higher among females than males. Multivariate analysis showed that male sex was a significant predictor of a negative treatment outcome, after controlling for other significant sociodemographic and health care related variables. PMID- 15748074 TI - Do private doctors follow national guidelines for managing pulmonary tuberculosis in Pakistan? AB - As private medical practitioners play a major role of in providing care to pulmonary tuberculosis (TB) patients, a survey was made of knowledge and practice in 2 cities in Pakistan. Only 1 of the 245 physicians was aware that cough > 3 weeks alone is the main symptom suggesting pulmonary TB. The majority diagnosed (80%) and treated (83%) cases themselves without referral. Less than 1% relied on sputum microscopy alone for diagnosis. None of the practitioners were following National TB Control guidelines for prescribing drugs and none ensured compliance with anti-TB treatment under supervision of a doctor/health worker. Only 3% kept records of pulmonary TB patients. None of the physicians assessed the effectiveness of treatment with sputum microscopy alone; the majority (76%) used only clinical assessment. PMID- 15748073 TI - Tuberculosis diagnosis and treatment practices of private physicians in Karachi, Pakistan. AB - In a densely populated urban area of Karachi, Pakistan, a questionnaire survey was made of the knowledge and practices of 120 private general practitioners about the diagnosis and treatment of tuberculosis (TB). The majority knew that cough, fever and weight loss were the main symptoms of TB, but less than half knew that blood in sputum, poor appetite and chest pain were associated with the disease. Only 58.3% of physicians used sputum microscopy for diagnosing TB and 35.0% used it as a follow-up test. Only 41.7% treated TB patients themselves, the remaining referring their patients to specialists. Around 73.3% of the doctors were aware of the 4 first-line anti-TB drugs. Efforts to improve the knowledge of private practitioners, and strategies to enhance public-private collaboration forTB control in urban areas are urgently required. PMID- 15748075 TI - Do doctors in north-western Somalia follow the national guidelines for tuberculosis management? AB - A cross-sectional study assessed the knowledge and practices of registered practitioners in management of tuberculosis (TB) in north-western Somalia. Of 100 registered doctors, 53 were interviewed. Of these, 32 (64%) had treated TB patients during the previous year, but only 1 had notified the authorities, 33 (66%) knew the most important symptoms and 32 (64%) identified sputum smear microscopy as the most important diagnostic test. Only 4 doctors prescribed the correct regimen and only 7 advocated direct observation. Suboptimal knowledge was more common among doctors working in private practice (relative risk: 2.1; 95% CI: 1.1-4.3). Patients are being treated in the private sector, but few doctors follow national treatment guidelines. Training in diagnosis and case management is needed to improveTB control in Somalia. PMID- 15748076 TI - Adherence of the private sector to national tuberculosis guidelines in the Islamic Republic of Iran, 2001-02. AB - We determined the knowledge and practice of private sector physicians in three cites regarding management of pulmonary tuberculosis (TB) and their determinants. A random sample questionnaire survey was conducted of 732 private physicians. Stratified analysis and logistic regression were used to identify the adjusted determinants of the two outcomes. A high proportion of the doctors had correct knowledge about the major diagnostic criteria but there was a low level of knowledge and practice of TB management. Significant risk factors for poor knowledge were age > or = 36 years, being a GP rather than a specialist and no attendance in TB training courses or attendance in courses held by institutions other than the public health system. Age > or = 36 years was the only significant risk factor for poor practice. PMID- 15748077 TI - Epidemiology of cutaneous leishmaniasis in the endemic area of Jericho, Palestine. AB - This study of cutaneous leishmaniasis in Jericho city and the adjacent Aqbat Jaber refugee camp investigated the seroprevalance of Leishmania major and the risk factors associated with acquiring the disease. Clinical and parasitology identification of cases showed children and young men were more affected, with the head most affected in children. Enzyme-linked immunosorbent assay (ELISA) was used to test sera from 190 individuals. The overall seroprevalence of cutaneous leishmaniasis was 26.3%. A case-control study of 247 individual in 37 households showed that a higher level of education of the head of the household and having children sleep under bed nets were significantly related to a lower incidence of cutaneous leishmaniasis. PMID- 15748078 TI - Epidemiological study in a new focus of cutaneous leishmaniasis in the Islamic Republic of Iran. AB - An outbreak of cutaneous leishmaniasis (CL) in Sabzevar county prompted this study of the epidemiology and the ecology of vectors and reservoirs. Examination of 541 schoolchildren showed rates of 9.4% for scars and 5.9% for ulcers. Among 807 inhabitants of 4 villages, 10.4% had scars and 3.0% had active lesions. The most highly infected age group was 0-4 years with a rate of 5.9%. A total of 12 849 sandflies representing 7 species were collected in the study area. Leptomonad infection was found in Phlebotomus papatasi, P. caucasicus and Sergentomyia sintoni. Parasites from man, P. papatasi and Rhombomys opimus, were isolated and characterized as Leishmania major. Based on this survey, this is an epidemic of zoonotic CL, with R. opimus the main reservoir host, and P. papatasi the main vector. PMID- 15748079 TI - Factors affecting variations in exposure to infections by Leishmania donovani in eastern Sudan. AB - A cross-sectional survey was carried out in Gedaref state, eastern Sudan to investigate the prevalence of positive leishmanin skin tests and environmental factors related to Leishmania donovani infection. A total of 3835 people living in 11 villages in 3 regions were screened. Soil types and tree densities were determined in 33 villages inhabited by 44 different tribes. The highest rates of positive skin tests were in Rahad region (33.9%), Atbara (21.6%) and Gedaref (10.6%), with an average of 21.1% for the state. Risk of infection by L. donovani varied significantly between different tribes. Higher densities of Acacia and Balanites spp. trees were in Masaleet villages, suggesting that the relatively high risk of L. donovani exposure in this tribe is due to environmental factors. PMID- 15748080 TI - Antimony-resistant Leishmania donovaniin eastern Sudan: incidence and in vitro correlation. AB - A longitudinal study was done in a leishmaniasis -endemic region in eastern Sudan during the period November 2001-February 2003 to determine the incidence of failure of sodium stibogluconate treatment. We studied 820 confirmed visceral leishmaniasis patients. All were treated with sodium stibogluconate, 20 mg/kg body weight for at least 28 days. Parasites were isolated from lymph node aspirates from 22 participants identified as relapsed patients. All isolates were typed as Leishmania donovanibased on polymerase chain reaction (PCR) amplification of parasite kDNA. Six parasites showed in vitro resistance to sodium stibogluconate using murine J774 macrophage amastigote testing method. The resistant isolates showed different restriction profiles when the amplified kDNA PCR products were digested with ALU1 restriction enzyme, indicating that resistance was mediated by different parasite clones. PMID- 15748081 TI - Field evaluation of latex agglutination test for detecting urinary antigens in visceral leishmaniasis in Sudan. AB - A latex agglutination test to detect urinary antigens for visceral leishmaniasis (VL) was studied. In 204 patients with suspected VL, KAtex had a sensitivity of 95.2% with good agreement with microscopy smears but poor agreement with 4 different serology tests. It was also positive in 2 confirmed VL cases co infected with HIV. In all K4tex-positive confirmed cases actively followed up after treatment, the test became negative 1 month after completion of treatment. While IC4tex had a specificity of 100% in healthy endemic and non-endemic controls, the direct agglutination test (DAT) was positive in 14% of the KAtex negative healthy endemic controls. KAtex is a simple addition to the diagnostics of VL particularly at field level and as a complementary test for the diagnosis of VL in smear-negative cases with positive DAT results. PMID- 15748082 TI - Visceral leishmaniasis in the Syrian Arab Republic: early detection using rK39. AB - Leishmaniasis causes significant morbidity and mortality in areas where it is endemic. A seroprevalence survey was conducted in 2 endemic villages in Daraa, Syrian Arab Republic, where 80 out of 345 children (23.2%) tested positive for visceral leishmaniasis (VL) using rK39 dipstick test. Only 10 cases were symptomatic (12.5%), and 27.5% were positive by ELISA test. All the sera (N = 138) obtained from the control village were negative. Of the rK39 initially positive cases, 52 had seroconverted to negative 9 months later, 55 remained ELISA negative, and none developed the full-blown disease. Being faster and less expensive than other diagnostic tests, rK39 is a rapid, sensitive and specific diagnostic tool for symptomatic cases of VL in remote areas with poor accessibility to health services. PMID- 15748083 TI - Filariasis elimination in Egypt: impact of low microfilaraemics as sources of infection for mosquitoes. AB - The elimination strategy for lymphatic filariasis aims at reducing blood microfilaraemia to levels at which vector transmission cannot be sustained. We aimed to determine whether patients with pre-treatment low or ultra-low microfilaria (MF) counts could be a reservoir of infection after mass drug administration (MDA) with a combined regimen. Laboratory-reared mosquitoes were fed on 30 volunteers after 2 rounds of MDA. Microfilaria uptake, infectivity rates and number of Wuchereria bancrofti L3 per mosquito were assessed. One year after MDA-1, 6 subjects transmitted MF, but up to 9 months after MDA-2 transmission failed. Six months after MDA-2 > 90% had clear MF smears and either failed to transmit MF or transmitted MF that did not develop to L3. We conclude that the transmission cycle is seriously weakened after MDA-2. PMID- 15748084 TI - Friction-cost method as an alternative to the human-capital approach in calculating indirect costs. PMID- 15748085 TI - The friction-cost method : replacement for nothing and leisure for free? AB - The friction-cost method has been put forward as an alternative to the human capital method as it allows more realistic estimates of productivity costs to be calculated for use in economic evaluations. The possibility of replacement of (long-term) absentees is at the heart of the friction-cost method. It recognises that society will restore initial production levels after some period of adaptation, the length of which may depend on the availability of labour and, hence, on unemployment. The friction-cost method has received two main criticisms in the literature: (i) it has no theoretical underpinning; and (ii) it treats leisure time as having no value. We demonstrate in a simple 'theoretical' time allocation model how time use shifts in the friction-cost method and that leisure is not treated as having no value. Rather, it is considered to be valued in terms of QALYs--as is normally the case in economic evaluation. The time-allocation model also demonstrates that when using the friction-cost or human-capital method the changes in the amount of unpaid work and leisure time need to be valued separately. Unpaid production losses from the previously unemployed may be larger than the gain in unpaid production gain of the absentee, resulting in a societal loss of unpaid work or the sacrifice of leisure in order to make up for lost unpaid work. These changes should be incorporated into economic analyses. PMID- 15748086 TI - The diffusion of health economics knowledge in Europe : The EURONHEED (European Network of Health Economics Evaluation Database) project. AB - This paper overviews the EURONHEED (EUROpean Network of Health Economics Evaluation Databases) project. Launched in 2003, this project is funded by the EU. Its aim is to create a network of national and international databases dedicated to health economic evaluation of health services and innovations. Seven centres (France, Germany, Italy, The Netherlands, Spain, Sweden and the UK) are involved covering 17 countries. The network is based on two existing databases, the French CODECS (COnnaissance et Decision en EConomie de la Sante) database, created in 2000 by the French Health Economists Association (College des Economistes de la Sante), and the UK NHS-EED (NHS Economic Electronic Database), run by the Centre for Reviews and Dissemination, University of York, York, England. The network will provide bibliographic records of published full health economic evaluation studies (cost-benefit, cost-utility and cost-effectiveness studies) as well as cost studies, methodological articles and review papers. Moreover, a structured abstract of full evaluation studies will be provided to users, allowing them access to a detailed description of each study and to a commentary stressing the implications and limits, for decision making, of the study. Access will be free of charge. The database features and its ease of access (via the internet: http://www.euronheed.org) should facilitate the diffusion of existing economic evidence on health services and the generalisation of common standards in the field at the European level, thereby improving the quality, generalisability and transferability of results across countries. PMID- 15748087 TI - The case for stratified cost-effectiveness analysis by baseline health-related QOL: theory and sensitivity analysis. AB - The purpose of this paper is to argue that systematic data heterogeneity exists when the objective of pharmacotherapy is improved health-related quality of life (HR-QOL), and that the pharmacotherapy's cost effectiveness will vary inversely with the patient's baseline HR-QOL (BHR-QOL), the patient's HR-QOL just prior to treatment. It is argued that when improved HR-QOL is the primary objective of a pharmacotherapy, the magnitude of the HR-QOL response to treatment may be negatively correlated with BHR-QOL and that the net cost of the pharmacotherapy may be positively correlated with BHR-QOL. It is also argued that the value placed on a given increment in HR-QOL declines as BHR-QOL rises. The case for sample stratification by BHR-QOL, and for conducting cost-effectiveness analysis (CEA) at the stratum level, is illustrated with numerical examples using hypothetical data and the incremental net monetary benefit (INMB) criterion. Sensitivity analysis is used to explore the response of the INMB at the stratum level to different degrees of data heterogeneity across the BHR-QOL strata. This paper demonstrates that because of cumulative effects, even relatively minor data heterogeneity related to BHR-QOL results in substantial differences in the cost effectiveness of treatments across BHR-QOL strata. CEA stratified by BHR-QOL enables a more efficient and equitable allocation of public healthcare funding than standard cost-effectiveness screening of pharmaceutical drugs based on full sample averages. PMID- 15748088 TI - Cost effectiveness of treating low HDL-cholesterol in the primary prevention of coronary heart disease. AB - BACKGROUND: A low serum level of high-density lipoprotein (HDL)-cholesterol is an independent risk factor for coronary heart disease (CHD). Fibrates, particularly gemfibrozil, have been shown to raise HDL-cholesterol levels and reduce the incidence of CHD. The literature on fibrate cost effectiveness is quite limited. OBJECTIVE: The objective of this analysis is to determine the cost effectiveness of the fibrates gemfibrozil and fenofibrate in the primary prevention of CHD. The target population includes patients with low levels of HDL-cholesterol, but without pre-existing CHD or other CHD risk factors sufficiently elevated to indicate drug therapy. STUDY DESIGN AND METHODS: From a societal perspective, a lifetime incremental cost-effectiveness model was developed to calculate baseline and treatment costs, life-years gained and QALYs gained. Model parameter values were taken from existing literature. In this 'backward induction' model, the expected costs and outcomes for each 5-year time-interval are utilised in subsequent 5-year time period calculations over the patient's entire lifetime. The study population consisted of a hypothetical cohort of males and females in the US aged 45-74 years, with low levels of HDL-cholesterol and no prior history of CHD. The base-case CHD risk factors for this population were obtained from the VA-HIT (Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial) population baseline characteristics, but assuming no prior CHD history. Estimates for the reduction in CHD risk associated with fibrate therapy reduction are also taken from the VA-HIT study. RESULTS: Using a societal cost-effectiveness threshold of US$50, 000 per QALY, primary prevention of CHD in patients with low HDL-cholesterol levels using generic gemfibrozil therapy is cost effective for all age and sex categories, in contrast to fenofibrate therapy, which is cost effective for males, but not for females at baseline risks levels. In the base case scenario, because of their higher CHD lifetime risk, it is more cost effective to treat males than females with either gemfibrozil or fenofibrate. For males and females the cost per QALY decreases with age for most age intervals. Gemfibrozil is more cost effective than fenofibrate for all age-sex categories because of the assumed equal efficacy and the higher fenofibrate drug cost. In the comparison scenario, generic lovastatin was more cost effective than gemfibrozil for men except at age 45 years and women at all ages, and more cost effective than fenofibrate for both men and women. CONCLUSIONS: This analysis suggests that fibrate therapy, particularly with generic gemfibrozil, is cost effective in the primary prevention of CHD in individuals with low HDL cholesterol levels, with or without elevated triglyceride levels. Certain patient subgroups, such as those with elevated triglyceride levels, smokers and those with diabetes mellitus are likely to achieve both CHD risk reduction and overall savings in net expected medical care costs. Comparable cost-effectiveness results are also shown for lovastatin therapy in the target patient population. Gemfibrozil dominates fenofibrate because of the lower cost of therapy (direct and indirect costs). These conclusions are robust to reasonable changes in model parameter values. PMID- 15748089 TI - Providing systematic guidance in pharmacoeconomic guidelines for analysing costs. AB - BACKGROUND: Despite the considerable amount of work that has gone into the standardisation of costs, a recent survey indicated that there is still a wide variation in methods used in costing studies in the published literature. Part of the answer to the lack of standardisation may lie in pharmacoeconomic guidelines themselves. OBJECTIVE: The purpose of this paper is to categorise the current recommendations for the analysis of costs within an economic evaluation, and to use these categories to assess variations in guidelines. DESIGN: Based on identified costing methodology, we developed a detailed categorisation. We then surveyed existing guidelines to determine similarities and differences in the guidelines. RESULTS: Our framework had five categories: general design items; resource identification and classification; resource measurement; resource valuation; and overall reporting. We noted differences in perspective, some of which were due to methodological uncertainty and differences in the purposes of the studies covered by the guidelines. There was very little guidance in how to put the basic cost measure into operation. In addition, the preferred concept for resource utilisation was that of resource use under actual (rather than experimental) conditions, and there was very little guidance as to how to achieve this measure. CONCLUSION: There are still large variations, as well as gaps, in guidelines in the costing area. This is due, in part, to wide-ranging objectives for studies, methodological uncertainty and a neglect of the fundamentals of costing. We recommend that the contents of the basic measure of 'average cost' should be explained in more detail, with recurring, capital and allocated overhead costs being reported separately. A target capacity level (e.g. 80%) should be reported, even if in a sensitivity analysis. More guidance should be provided regarding the translation of utilisation under experimental care to that under routine care. For resource valuation, adjustments to the 'average cost' measure should be made explicit. There should be more guidance on how to project system-wide costs from the costs of one hospital. The manner in which overall results are reported should be addressed, including the identification of how costs should be categorised. PMID- 15748090 TI - Evaluation of the cost effectiveness of escitalopram versus venlafaxine XR in major depressive disorder. AB - AIM: To assess the relative cost effectiveness of escitalopram compared with venlafaxine XR in patients with major depressive disorder (MDD). METHODS: An economic evaluation was conducted alongside a double-blind, multinational, randomised clinical trial and examined the costs and quality of life of 251 patients taking escitalopram versus venlafaxine. Outpatients fulfilling criteria for MDD were randomised to receive oral escitalopram 10-20 mg/day or venlafaxine 75-150 mg/day for 8 weeks. Patient-reported outcomes (EuroQOL questionnaire, Quality of Life Depression Scale), use of medical services and absence from work (relating to the previous 3 months) were recorded at baseline, with repeated measurements at week 8. Unit costs in year values were applied to the resource utilisation data. A cost-effectiveness analysis was performed using the EuroQOL score as the effectiveness measure. The perspective was that of the healthcare payer, with a societal perspective considered in a sensitivity analysis. RESULTS: Statistically significant improvements in patient-reported outcomes (vs baseline) were observed in both groups after 8 weeks' treatment. Patients treated with escitalopram tended to report fewer problems on the EuroQOL dimensions than venlafaxine recipients. Mean per-patient costs in euros (euro, year 2003 values) for the escitalopram group, compared with the venlafaxine group, were 32% lower (110 euros vs 161euros) from a healthcare perspective, although this was not a statistically significant difference. Differences were related to lower drug acquisition costs and fewer hospitalisations for escitalopram than venlafaxine recipients. A multivariate model adjusting for baseline characteristics showed that escitalopram reduced direct costs compared with venlafaxine (p = 0.007). Bootstrapped distributions of the incremental cost-effectiveness ratios also showed similar effectiveness but lower costs for escitalopram compared with venlafaxine. Inclusion of indirect costs led to similar results. CONCLUSION: This prospective economic analysis suggests that escitalopram has similar effectiveness to venlafaxine in the treatment of MDD, but may be associated with lower healthcare costs. These findings are consistent with previously published economic evaluations. PMID- 15748091 TI - Cost-minimisation analysis of sivelestat for acute lung injury associated with systemic inflammatory response syndrome. AB - OBJECTIVES: To conduct a cost-minimisation analysis of sivelestat sodium hydrate treatment for patients in the intensive care unit (ICU) with acute lung injury (ALI) associated with systemic inflammatory response syndrome (SIRS) caused by infection. DESIGN: The analysis was performed based on data from a phase III randomised, multicentre, double-blind, controlled clinical study of up to 14 days treatment with sivelestat, in which the effect of intravenous sivelestat at a high dose (0.20 mg/kg/h; the sivelestat group) was compared with that at a low dose (0.004 mg/kg/h, effectively a placebo; the control group). PATIENTS: Patients with ALI associated with SIRS caused by infection, who began their treatment under mechanical ventilation management in the ICU. METHODS: A four stage Markov model was constructed to represent the possible conditions of an ALI patient: ICU plus intubated mechanical ventilation; ICU plus weaned from a mechanical ventilator; admission to the general ward; and death. The base-case analysis used a mechanical ventilator weaning daily rate of 2.9% for the control group and 4.0% for the sivelestat group, and the same mortality (1.2%) for both groups at all stages of the Markov model. Medical costs were estimated from standard fees and Japanese National Health Insurance drug prices included fees for hospitalisation within the ICU and general wards, mechanical ventilation, examinations and drug expenditure. Costs were in 2001 values. Sensitivity analyses were performed by varying the weaning rate, mortality, time between weaning and discharge to the general ward, and drug costs. PERSPECTIVE: Payers of healthcare costs. MAIN OUTCOMES: The expected 30-day medical costs per patient in the control and sivelestat groups were Japanese yen (yen) 4,144,887 and 3,975,451 yen, respectively; a difference of 169,436 yen. Drug expenditure accounted for more than half of the medical costs for each group. The periods under mechanical ventilation management and in the ICU for the sivelestat group were shorter than those for the control group by 2 and 1.8 days, respectively. This was of significance in the reduction of the medical costs. A sensitivity analysis suggested that the expected costs for the sivelestat group exceeded those for the control group when the daily weaning rate for the sivelestat group was <3.5%, and also when mortality rates were set at 0.9% in the sivelestat group and 1.4% in the control group. CONCLUSIONS: This analysis suggests that from the Japanese healthcare payer perspective, treatment with sivelestat may reduce medical costs compared with standard care for patients with ALI associated with SIRS caused by infection. PMID- 15748092 TI - The impact of anaemia and its treatment on employee disability and medical costs. AB - OBJECTIVE: Anaemia is a common haematological complication of cancer and cytotoxic treatment. The incremental economic burden associated with medical care and short-term disability of anaemia in patients with malignancy and receiving chemotherapy has not been well documented. This study evaluates the effect of anaemia on the costs associated with cancer treated with chemotherapy. METHODS: Patients receiving chemotherapy within 6 months of their initial cancer diagnosis were identified in a commercial claims database for 1999-2000. Data for these individuals were linked to their employers' short-term disability records via unique encrypted personal identification numbers provided by employers. Patients with anaemia were identified by a diagnosis of anaemia or treatment with transfusion or erythropoietin alfa (EPO). Healthcare expenditure and short-term disability leave were observed for up to 6 months following initial cancer diagnosis and were summarised into monthly averages. Exponential conditional mean models and zero-inflated negative binomial models were used to analyse mean monthly healthcare expenditures and short-term disability days. RESULTS: Twenty five percent of the 619 newly diagnosed cancer patients treated with chemotherapy had anaemia. The presence of anaemia and longer length of transfusion therapy were associated with increased expenditures, while longer length of EPO treatment was associated with lower expenditures. The incremental costs due to anaemia among patients receiving chemotherapy were US$5,538 (year 2001 values) per month in the first 6 months following cancer diagnosis, 10.8% of which were costs related to short-term disability leave. CONCLUSION: Anaemia in patients undergoing chemotherapy presents a substantial burden to employers and payers. The findings also suggest that patients with anaemia treated with erythropoietin alfa can achieve expenditure levels similar to those patients without anaemia. PMID- 15748093 TI - Memantine: a pharmacoeconomic review of its use in moderate-to-severe Alzheimer's disease. AB - Memantine (Ebixa, Namenda, Axura) is an uncompetitive NMDA receptor antagonist used in the management of patients with moderate-to-severe Alzheimer's disease. It is currently the only drug approved for use in these more advanced stages of the disease. Significant reductions in functional and cognitive decline have been demonstrated with memantine relative to placebo in randomised, double-blind trials in this patient population. Clinical trial and postmarketing surveillance data indicate that the drug is generally well tolerated. Two fully published modelled cost-effectiveness analyses of memantine in moderate-to-severe Alzheimer's disease have been conducted in the UK and Finland, in which patient progression was simulated through health states related to dependency, residential setting and cognitive function. Although the specific costs included in the analyses varied, as did the study perspective and geographical location, results of the base-case analyses consistently showed that memantine was dominant over no pharmacological treatment. In the UK and Finnish analyses, memantine increased the duration of independence by 1.3 and 4.1 months, respectively, and the time to institutionalisation by 0.8 and 1 month. Mean total per-patient costs were reduced by 1963 pounds over 2 years (2003 costs) in the UK analysis and by 1687 eurossover 5 years (2001 costs) in the Finnish analysis. Memantine was also associated with a small gain in quality-adjusted life expectancy in the UK model. In sensitivity analyses, memantine remained dominant for almost all plausible changes to key variables. Memantine reduced total societal costs by $1090 per patient per month (1999 costs) compared with no pharmacological treatment over 28 weeks in a resource utilisation and cost analysis conducted alongside a pivotal US trial in patients with moderate-to-severe Alzheimer's disease. Results were primarily driven by reductions in total caregiver costs, which included the opportunity cost of time spent in caregiving tasks, and in direct nonmedical costs, which included the cost of care in a nursing home or similar institution.In conclusion, in patients with moderate-to-severe Alzheimer's disease, memantine is associated with significant reductions in functional and cognitive decline compared with no pharmacological treatment. Available pharmacoeconomic data from Europe and the US, despite some inherent limitations, support the use of memantine as a cost-effective treatment in this patient population, although definitive conclusions are not feasible because of limited data. PMID- 15748095 TI - Contraceptive vaccines. AB - The world's population is growing at a tremendous rate, affecting growth and development. Apart from this population growth, unintended pregnancies resulting in elective abortions continue to be a major public health issue. In over half of these unintended pregnancies, the women have used some type of contraception. Thus, there is an urgent need for a better method of contraception that is acceptable, effective and available. The contraceptive choices available to women at this time include steroid contraceptives, intrauterine devices, barrier methods, spermicides, natural family planning, male and female sterilisation, and recently available emergency contraceptives. Contraceptive vaccines (CVs) may provide viable and valuable alternatives that can fulfill most, if not all, properties of an ideal contraceptive. Since both the developed and most of the developing nations have an infrastructure for mass immunisation, the development of vaccines for contraception is an exciting proposition. The molecules that are being explored for CV development either target gamete production (gonadotropin releasing hormone, follicle-stimulating hormone and luteinising hormone), gamete function (zona pellucida [ZP] proteins and sperm antigens) or gamete outcome (human chorionic gonadotropin [hCG]). Disadvantages of CVs targeting gamete production are that they affect sex steroids and/or show only a partial effect in reducing fertility. CVs targeting gamete function are better choices. Vaccines based on ZP proteins are quite efficacious in producing contraceptive effects. However, they invariably induce oophoritis affecting sex steroids. Sperm antigens constitute the most promising and exciting targets for CVs. Several sperm specific antigens have been delineated in several laboratories and are being actively explored for CV development. Antisperm antibody-mediated immunoinfertility provides a naturally occurring model to indicate how an antisperm vaccine will work in humans. Vaccines targeting gamete outcome primarily focus on the hCG molecule. The hCG vaccine is the first vaccine to undergo phase I and II clinical trials in humans. Both the efficacy and the lack of immunotoxicity have been reasonably well demonstrated for this vaccine. The present studies focus on increasing the immunogenicity and efficacy of this birth control vaccine. PMID- 15748096 TI - Treatment of Legionnaires' disease. AB - Legionnaires' disease is pneumonia, usually caused by Legionella pneumophila, which can range in severity from mild to quite severe. While it is commonly acquired in the community, it can just as easily be acquired nosocomially from water sources that have not been appropriately decontaminated. While historically initial treatment was always with erythromycin, current case series and treatment recommendations suggest that outpatients receive immediate treatment with one of the following antibacterials: azithromycin, erythromycin, clarithromycin, telithromycin, doxycycline or an extended-spectrum fluoroquinolone. If the symptoms are severe enough to warrant hospitalisation then the patient should receive treatment with parenteral azithromycin or extended-spectrum fluoroquinolones followed by step-down to oral formulations to complete the regimens. While a shorter course of 7-10 days for more severe infections may be possible for intravenous/oral azithromycin, other antibacterials should be administered for a total of 10-21 days and started as soon as possible upon presentation to optimise outcomes. PMID- 15748094 TI - Inhaled corticosteroids in chronic obstructive pulmonary disease: is there a clinical benefit? AB - Chronic obstructive pulmonary disease (COPD) is a serious illness that affects over 5% of the adult population. It is one of the few conditions for which the mortality and morbidity are still increasing. Experts expect COPD to become the third leading cause of death and the fifth leading cause of disability worldwide by the year 2020. Thus far, the only treatments that have been shown to make a difference to survival are smoking cessation and the use of oxygen supplements for those who are hypoxaemic at rest. The use of inhaled corticosteroids as monotherapy or in combination with a long-acting beta2-adrenoceptor agonist for COPD is controversial. Experimental data indicate that the inflammatory process in COPD may be resistant to the anti-inflammatory effects of corticosteroids. However, several large clinical studies have shown that inhaled corticosteroids in relatively high doses (e.g. budesonide 800 microg/day or fluticasone propionate 1 mg/day) reduce exacerbations by 20-30% and improve the health status of COPD patients by a similar amount compared with placebo. Withdrawal of inhaled corticosteroids may increase clinical exacerbation rates by 50% in COPD patients and by 2-fold in those with severe disease. Combined therapy with inhaled corticosteroids and long-acting beta2-adrenoceptor agonists may be superior to individual component therapy in reducing exacerbations. However, these medications must be used cautiously, as they have been associated with certain adverse effects. Inhaled corticosteroids, for instance, increase the risk for dysphonia and oral thrush by 2- to 3-fold. Skin bruising is also more common in users than in non-users of inhaled corticosteroids. On balance, for those with moderate-to-severe COPD and those who experience frequent exacerbations, judicious use of inhaled corticosteroids alone or in combination with long-acting beta2-adrenoceptor agonists appears reasonable. PMID- 15748097 TI - Combating chronic renal allograft dysfunction : optimal immunosuppressive regimens. AB - Kidney transplantation is the best treatment for patients with end-stage renal disease, both in terms of survival benefit and quality of life. The major limitation is the continuing shortage of kidneys suitable for transplantation, reinforcing the need to maximise graft survival. After the first year of transplantation, chronic renal allograft dysfunction (CRAD) is the first cause of late graft deterioration and failure. CRAD has been defined as a progressive renal dysfunction, independent of acute rejection, drug toxicity and recurrent or de novo nephropathy, with features on biopsy of chronic allograft nephropathy (CAN) characterised by vascular intimal hyperplasia, tubular atrophy, interstitial fibrosis and chronic transplant glomerulopathy. Protocol biopsy based studies have demonstrated a high and early prevalence of CAN lesions during the first year in patients with normal and stable renal function. Beyond 1 year, the injuries associated with calcineurin inhibitors (CNIs) appear to be very common. The physiopathology of CRAD is complex and multifactorial. Both alloantigen-dependent factors (acute rejection, HLA matching, donor-specific antibodies, inadequate immunosuppression) and alloantigen-independent factors (donor age, brain death, ischaemia/reperfusion injuries, hypertension, hyperlipidaemia, cytomegalovirus, CNI-related nephrotoxicity) are involved. Consequently, CRAD appears as a dynamic process, evolving with time, and immunosuppressive regimens need to be modulated in order to provide the most suitable treatment at the different phases of its natural history. On the basis of this scheme, the new paradigm would be the use of a CNI-based regimen during the period of maximal risk of (subclinical) acute rejection, followed by a conversion to a CNI-free regimen in order to avoid the long-term consequences of nephrotoxicity. Fortunately, new agents are being introduced in clinical practice providing a large range of combinations and allowing individualisation of immunosuppressive regimens. Large, prospective, multicentre trials are warranted, and the challenge is to define new endpoints of CRAD and to determine the best therapeutic strategy. PMID- 15748098 TI - Amprenavir or fosamprenavir plus ritonavir in HIV infection: pharmacology, efficacy and tolerability profile. AB - Amprenavir is an HIV-1 protease inhibitor, the first in vitro activity studies of which were published in 1995. During in vivo development, it became clear that the pharmacokinetics of the drug would result in patients taking a large number of pills daily. The first comparative studies of amprenavir versus other protease inhibitors showed it had comparatively weak activity. Thus, studies using low doses of ritonavir to enhance the pharmacokinetic profile of amprenavir were first communicated in 2000. Only a small number of clinical trials in HIV-1 infected patients have been published. The pharmacokinetics of amprenavir have been documented in both healthy individuals and in HIV-1-infected patients. Amprenavir trough plasma concentrations increase 3- to 10-fold and the area under the concentration-time curve (AUC) increases 2- to 3-fold when using amprenavir 450 or 600 mg combined with ritonavir 100mg twice daily. Peak concentrations of amprenavir are less influenced by ritonavir coadministration, with a 1- to 2-fold increase. As there is no pharmacokinetic advantage to increasing ritonavir doses, the combination has only been used with low doses of ritonavir (100mg twice daily or 200 mg once or twice daily). Concomitant use of currently available non nucleoside reverse transcriptase inhibitors (NNRTIs)--efavirenz or nevirapine--is possible when amprenavir is coadministered with ritonavir, despite the pharmacokinetic interactions described when they are used with amprenavir alone. Fosamprenavir (GW 433908) is a prodrug of amprenavir primarily metabolised to amprenavir in the epithelial cells of the intestine. At steady state, plasma trough concentrations and AUC are slightly greater with fosamprenavir (two pills of 700 mg twice daily) than amprenavir (eight soft gel capsules of 150 mg twice daily). The clinical adverse effects of amprenavir are similar whether administered unboosted or in combination with ritonavir. Skin rashes do not appear to be more frequent. With regard to lipid profiles, the addition of ritonavir to amprenavir induces an increase in cholesterol and triglyceride levels; however, prospective comparative studies are lacking. In short-term prospective trials in antiretroviral-naive individuals, virological suppression with highly active antiretroviral therapy containing amprenavir plus ritonavir is similar to or higher than with unboosted amprenavir, with a smaller pill intake. Few comparative data are available in treatment-experienced patients. In several small studies, different salvage regimens which included amprenavir plus ritonavir achieved undetectable viral levels in half of the patients. Although the I50V amino acid substitution is the key mutation conferring resistance to amprenavir, the accumulation of several mutations is needed to increase the IC50 (concentration that produces 50% inhibition) of amprenavir. When used with ritonavir, the accumulation of six or more mutations among L10F/I/V, K20M/R, E35D, R41K, I54V, L63P, V82A/F/T/S and I84V leads to clear decrease in viral response to treatment. In salvage regimens, coadministration of amprenavir with lopinavir/ritonavir induces variations in lopinavir and amprenavir concentrations (decrease or increase in both drug concentrations) compared with the combination with ritonavir alone. Currently, close pharmacokinetic follow-up is mandatory when such combinations are used. There are sufficient data available today to support coadministration of reduced doses of amprenavir with low doses of ritonavir. Compared with amprenavir alone, this results in the administration of fewer pills with equivalent or higher efficacy, but without new clinical adverse effects. The concentrations of amprenavir achieved are high enough for use in treatment-experienced patients who have an accumulation of amino acid substitutions in the HIV-1 protease gene. It also allows combinations with NNRTIs. The pharmacokinetic properties of fosamprenavir and the first clinical trials in treatment-naive and treatment-experienced patients should allow it to be considered as a better alternative to amprenavir in countries where fosamprenavir is already available. PMID- 15748101 TI - A new three-dimensional method of assessing facial volumetric changes after orthognathic treatment. AB - OBJECTIVE: To validate a new method of facial volumetric assessment that is dependent on the use of stereophotogrammetric models and a software-based Facial Analysis Tool. DESIGN: The method was validated in vitro with three-dimensional (3D) models of a lifelike plastic female dummy head and in vivo with a male subject head. METHODS: Thirty facial silicone explants were added in the nasal and perioral regions of each head, and their volumes were obtained by three different algorithms. These were compared with the actual values obtained by a "water displacement" method. RESULTS: The least mean error was found with the "tetrahedron formation" method followed by the "projection" method and the "back plane construction" method. The error with the tetrahedron formation method was 0.071 cm(3) (95% confidence interval [CI]: -0.074 to 0.2161 cm3) with the in vitro models and 0.314 cm3 (95% CI: -0.080 to 0.708 cm3) with the in vivo models. The increased volumetric assessment error observed in vivo was attributed to the registration procedure and possible changes in facial expression. CONCLUSIONS: These results encourage the use of this method in the 3D assessment of orthognathic surgical outcome, provided a standardized facial expression is used for image acquisition. PMID- 15748100 TI - Gatifloxacin: a review of its use in the treatment of bacterial infections in the US. AB - Gatifloxacin (Tequin) is an 8-methoxy fluoroquinolone approved in the US for use in the treatment of community-acquired pneumonia (CAP), acute exacerbations of chronic bronchitis (AECB), acute sinusitis, uncomplicated and complicated urinary tract infections (UTIs), pyelonephritis, gonorrhoea and uncomplicated skin and skin structure infections. Gatifloxacin has a broad spectrum of antibacterial activity in vitro and good clinical and bacteriological efficacy in patients with indicated infections following once-daily administration by the intravenous or oral routes. It is generally well tolerated; the most common adverse events are associated with the gastrointestinal tract and CNS. Recent approvals for the use of gatifloxacin in the treatment of CAP due to multidrug-resistant Streptococcus pneumoniae (MDRSP) and in uncomplicated skin and skin structure infections extend the role of this drug in the treatment of bacterial infections in the US. PMID- 15748102 TI - A new protocol for maxillary protraction in cleft patients: repetitive weekly protocol of alternate rapid maxillary expansions and constrictions. AB - OBJECTIVE: It was hypothesized that, through a repetitive weekly protocol of Alternate Rapid Maxillary Expansions and Constrictions (Alt-RAMEC), the maxilla in cleft patients could be protracted more effectively than with a single course of rapid maxillary expansion (RME). METHODS: Twenty-six consecutive unilateral cleft lip and palate patients at the age of mixed dentition were included in this prospective clinical study. The rapid maxillary expansion group included the first 16 consecutive patients undergoing 1 week of rapid maxillary expansion (1 mm/day) followed by 5 months, 3 weeks of maxillary protraction. The Alternate Rapid Maxillary Expansions and Constrictions group included the next 10 consecutive patients undergoing 9 weeks of Alternate Rapid Maxillary Expansions and Constrictions followed by 3 months, 3 weeks of maxillary protraction. Daily activation of the weekly expansion or constriction was 1.0 mm. Two-hinged expanders and intraoral maxillary protraction springs were used in both groups. Treatment results were evaluated cephalometrically. RESULTS: The amount of maxillary anterior displacement by the 2-hinged expander in the Alternate Rapid Maxillary Expansions and Constrictions group was 3.0 +/- 0.9 mm at A point, significantly greater than the 1.6 +/- 1.0 mm in the rapid maxillary expansion group. The amount of maxillary advancement with intraoral protraction springs in the Alternate Rapid Maxillary Expansions and Constrictions group was 2.9 +/- 1.9 mm at A point, significantly greater than the 0.9 +/- 1.1 mm in the rapid maxillary expansion group. The overall amount of maxillary advancement in the Alternate Rapid Maxillary Expansions and Constrictions group was 5.8 +/- 2.3 mm at A point. This result remained stable, without significant relapse after 2 years. CONCLUSIONS: Maxillary protraction using the 2-hinged expander, a repetitive weekly protocol of Alternate Rapid Maxillary Expansions and Constrictions, and intraoral protraction springs is most effective, with stable results at 2-year follow-up. PMID- 15748099 TI - Efficacy, tolerability and cost effectiveness of disease-modifying antirheumatic drugs and biologic agents in rheumatoid arthritis. AB - Over the last decade, several new drugs have become available for the treatment of patients with rheumatoid arthritis. These agents include the new disease modifying antirheumatic drug (DMARD) leflunomide and the biologic agents, tumor necrosis factor (TNF)-alpha antagonists and an interleukin (IL)-1 receptor antagonist. Methotrexate is commonly used as the first DMARD, has a well documented clinical efficacy and slows radiological deterioration. Sulfasalazine appears to have similar properties, albeit to a lesser extent. Leflunomide has similar efficacy as methotrexate but it is less tolerated than sulfasalazine. The adverse effect profiles of these three drugs makes regular laboratory monitoring mandatory. Several combination therapies with DMARDs were proven to be more effective than mono-DMARD therapy. However, until now these strategies have not been widely adopted. TNF antagonists are potent anti-inflammatory drugs, with a rapid onset of effects compared with traditional DMARDs. The IL-1 receptor antagonist, anakinra, has an intermediate place between methotrexate and the TNF antagonists with respect to efficacy. The adverse effects of TNF antagonists include an increased incidence of common and opportunistic infections. Thus far, anakinra has not been associated with an enhanced rate of opportunistic infections. Some of the biologic agents have been associated with worsening heart failure and demyelinating disease. The limited long-term safety data of the biologic agents are a point of concern because, at present, an enhanced risk for malignancies, particularly lymphoma, can not be excluded. Drug costs of traditional DMARDs are up to US dollars 3000 per year, whereas for the biologics the yearly drug costs range between US dollars 16,000 and > US dollars 20,000. Cost-effectiveness analyses are necessary to determine whether or not these high costs are justified. Unfortunately, adequate, prospective, economic evaluations are not yet available. Until these become available, treatment decisions will be based on the balance of direct costs and indirect costs and expected cost savings in the future. PMID- 15748103 TI - Application of limited cone beam computed tomography to clinical assessment of alveolar bone grafting: a preliminary report. AB - OBJECTIVES: The aim of this study was to demonstrate the clinical applicability of limited cone beam computed tomography (Dental 3D-CT) for assessment of bone grafted alveolar cleft. PATIENTS AND METHODS: Seventeen bone bridges were examined after alveolar bone grafting in 13 patients with cleft lip and palate. All bone bridges, including cleft-adjacent teeth, were examined by plain radiography and the Dental 3D-CT imaging system. RESULTS: The plain radiographs showed the approximate condition of the bone bridge and cleft-adjacent teeth. The Dental 3D-CT images clearly showed precise three-dimensional (3D) morphology of the bone bridge, 3D relationships between the bone bridge and the roots of cleft adjacent teeth, and their periodontal condition. In addition, the conditions surrounding dental implants installed in the bone bridge could be observed three dimensionally. CONCLUSIONS: The results indicate that the Dental 3D-CT imaging system is suitable for clinical assessment of alveolar bone grafting before and after installation of dental implants or orthodontic treatment of the cleft adjacent teeth. PMID- 15748104 TI - Ventral frontal cortex morphology in adult males with isolated orofacial clefts: relationship to abnormalities in social function. AB - OBJECTIVE: In a previous magnetic resonance imaging (MRI) study, men with nonsyndromic clefts of the lip and/or palate (NSCLP) were found to have abnormalities in the structure of the frontal lobe of the brain. Moreover, many subjects with nonsyndromic clefts of the lip and/or palate have been described as being socially inhibited. A subregion of the frontal lobe, the ventral frontal cortex (VFC), has been shown to be related to social function. This study was designed to evaluate the morphology of the ventral frontal cortex in men with nonsyndromic clefts of the lip and/or palate, and the morphology's relationship to social function. METHODS: Subjects were 46 men with nonsyndromic clefts of the lip and/or palate and 46 sex matched controls. Social function was assessed using a standardized scale. The morphology of the ventral frontal cortex (composed of the orbitofrontal cortex [OFC] and the straight gyrus [SG]) was obtained from magnetic resonance imaging scans using the software BRAINS. RESULTS: After controlling for frontal lobe gray matter, the patient group had significant reductions in orbitofrontal cortex volume and area. The straight gyrus was not morphologically abnormal. Measures of orbitofrontal cortex morphology were significantly correlated to measures of social function--the greater the structural abnormality, the greater the social dysfunction. CONCLUSION: Compared with healthy controls, subjects with nonsyndromic clefts of the lip and/or palate showed morphologic abnormalities in the cortical surface anatomy of a brain region known to govern social function, the orbitofrontal cortex. Moreover, the structural abnormality in this brain region was directly correlated with social function. PMID- 15748105 TI - Methods of assessment of cleft-related facial deformity: a review. AB - OBJECTIVE: To review the stimulus media, measurements and scoring system, rater's characteristics, and facial region used in assessment of the appearance of cleft related deformity. METHODS: A review of the literature was conducted using MedLine from January 1966 to June 2003. Key orthodontic journals and The Cleft Palate-Craniofacial Journal were hand searched as well. RESULTS: Most studies used a static two-dimensional means of assessment. Direct clinical assessment, three-dimensional media, or both have been used to a lesser extent. Only one study utilized all three assessment media. Three studies, however, attempted to compare the direct clinical assessment with that obtained using two-dimensional media. Most of the studies demonstrated that appearance of the cleft-related deformity could be measured by a variety of judges. The reliability and validity of the three different media as methods of assessment have received limited attention. The facial area assessed has varied from the full face, nasolabial area, or specific features of the lip or nose. CONCLUSIONS: Wide variation exists in the study design related to assessment of cleft-related deformity. An internationally agreed objective method of assessment for this facial deformity is required. A three-dimensional, rather than a two-dimensional, means of assessment should be adopted. PMID- 15748106 TI - Comparison of plaster casting with three-dimensional cranial imaging. AB - OBJECTIVE: The development of a new cranial imaging system to capture a three dimensional (3D) model of an infant's head has been previously reported. The accuracy of this new system has been independently established. However, before replacing the traditional plaster casting technique, the two methods require a comparison to ensure that the models they produce are equivalent. METHODS: Ten sequential infants were digitized by the 3D imaging system and the plaster casting technique following previously reported protocols. The cast models were also digitized so they could be compared with the digitized images. The two models (3dImage and Cast) were then imported into dimensional analysis software and aligned and registered with well-established registration algorithms. Difference maps that identified the variation between the two surfaces were generated for each pair, and descriptive statistics of these differences were recorded. RESULTS: The mean difference between the cast and the digitized models was 0.052 mm (standard deviation = 0.988), with a root mean square (RMS) difference of 1.028 mm. Two-dimensional slices obtained from the registered 3D surfaces demonstrated excellent agreement between the cranial contours. CONCLUSIONS: In this investigation, the 3D models created by the new imaging system were found to be within 1.0 mm RMS of the models created by the plaster casting technique. Two-dimensional cranial contours demonstrated excellent agreement between the two methods. The results of this investigation confirmed that the new cranial imaging system and the traditional plaster casting technique yield equivalent models. PMID- 15748107 TI - Human prenatal nasal bone lengths: normal standards and length values in fetuses with cleft lip and cleft palate. AB - OBJECTIVE: The aim of this study was to present prenatal autopsy standards for nasal bone length in normal fetuses, as well as to compare nasal bone lengths in human fetuses with cleft lip and palate with those standards. MATERIAL AND METHODS: The material consisted of human fetuses investigated in connection with diagnosed and legally approved abortions and spontaneous abortions; 40 were normal and 26 had cleft lip and palate (7 isolated cleft lip, 12 isolated cleft palate, and 7 combined cleft lip and palate). Menstrual ages (MA) ranged from 12 to 25 weeks, crown-rump length (CRL) from 55 to 210 mm and foot length (FL) from 7 to 44 mm. All fetuses were radiographed in lateral projection, and, using these images, the nasal bone lengths were measured with a digital caliper. Regression analyses were performed. RESULTS: Standards for normal nasal bone length at different ages are expressed as menstrual age, crown-rump length, or foot length. Comparisons with normal standards showed that the isolated cleft lip group had a significantly smaller nasal bone length than the normal fetuses had. Fetuses with isolated cleft palate and fetuses with combined cleft lip and palate showed no significant deviation from normal standards. CONCLUSION: Normal nasal bone standards can contribute to information on deviations in nasal bone length for aborted fetuses with known and unknown genotypes. The differences in nasal bone lengths in fetuses with different lip and palate clefting can contribute to understanding the pathogenesis of cleft lip and palate. PMID- 15748108 TI - Infant orthopedics in UCLP: effect on feeding, weight, and length: a randomized clinical trial (Dutchcleft). AB - OBJECTIVE: To study the effects of infant orthopedics (IO) on feeding, weight, and length. DESIGN: Prospective two-arm randomized controlled trial in three academic Cleft Palate Centers. Treatment allocation was concealed and performed by means of a computerized balanced allocation method. SETTING: Cleft Palate Centers of Amsterdam, Nijmegen, and Rotterdam, the Netherlands. PATIENTS: Infants with complete unilateral cleft lip and palate (UCLP), no other malformations. INTERVENTIONS: One group (IO+) wore passive maxillary plates during the first year of life, but the other group (IO-) did not. All other interventions were the same for both groups. MAIN OUTCOME MEASURES: Bottle feeding velocity (mL/min) at intake, 3, 6, 15, and 24 weeks (T0 to T24); weight-for-age, length-for-age, and weight-for-length using z scores; reference values from the Netherlands' third nationwide survey on growth. RESULTS: Feeding velocity increased with time from 2.9 to 13.2 mL/min in the IO- group and from 2.6 to 13.8 mL/min in the IO+ group; no significant differences were found between groups. Weight-for-age, length-for age, and weight-for-length (z scores) did not differ significantly between groups, but overall the infants with unilateral cleft lip and palate in both groups had significantly lower mean z scores for weight-for-age and height-for age than the reference during the first 14 months, and had lower mean values for weight-for-length after soft palate closure. CONCLUSION: Infant orthopedics with the aim of improving feeding and consequent nutritional status in infants with unilateral cleft lip and palate can be abandoned. PMID- 15748110 TI - Family environment of individuals with oral clefts in Argentina. AB - OBJECTIVE: The purpose of this investigation was to study the social environment of families of children with different types of nonsyndromic oral clefts (OC) and to compare these groups with a control population of families of children without clefts. DESIGN: The study compared three nonsyndromic oral cleft groups and the control group using the Moos Family Environment Scale, which examines cohesion, expressiveness, conflict, independence, achievement-orientation, intellectual cultural orientation, active-recreational orientation, moral-religious emphasis, organization, and control. SETTING: All parents of children with nonsyndromic oral clefts from a large craniofacial clinic in Buenos Aires, Argentina, were identified and were enrolled in this study between June 2000 and August 2001. Control families were ascertained from the pediatrics service of a hospital located in the vicinity of the craniofacial clinic. PARTICIPANTS: One hundred and sixty-five parents were selected, based on having a child with nonsyndromic unilateral cleft lip with or without cleft palate (UCL/P), bilateral cleft lip with or without cleft palate (BCL/P), or isolated cleft palate (ICP). One hundred and eighty control parents with no family history of congenital anomalies were selected, as well. RESULTS: There was no major difference in the social environment of families of children with different types of nonsyndromic oral clefts. When compared with families in the control group, families of children with nonsyndromic oral clefts scored better in all three subdimensions of family relationship, revealed a high level of independence, and showed better structure and organization than control families did; however, families of children with nonsyndromic oral clefts reported participating in fewer recreational activities. CONCLUSIONS: Overall, families of children with nonsyndromic oral clefts displayed a good social environment. Efforts should be focused to involve them in recreational activities. PMID- 15748109 TI - Report of altered sensation in patients with cleft lip. AB - OBJECTIVE: To determine whether patients with cleft lip have normal perioral sensation. DESIGN: Each subject was carefully questioned about the following: sensation in the face at rest, light touch of different areas, and sensation in natural situations (e.g., exposure to cold weather) that reveal sensory abnormalities. A cotton-tip applicator stick was stroked lightly across the facial skin. The subject's descriptions of the evoked sensations were used to identify and outline areas with abnormal sensation. SETTING: Data were obtained from subjects participating in a longitudinal, university-based study of the functional outcomes of lip revision surgery. PATIENTS: Seventeen patients with cleft lip and 12 control subjects (aged 7 to 22 years, mean 12.9 years) participated. RESULTS: In contrast to control subjects, 9 of 16 patients (56%) reported loss in sensation, described as decreased touch, scratch, tickle, or tingle intensity. Six other patients (38%) reported additional sensation, described as increased scratch, tickle, or tingle intensity. In eight patients, the altered sensation was restricted to the skin area flanking and including the visible scar, encompassing no more than 25% of the total area bound by the inferior nose, nasolabial grooves and inferior vermilion. In seven patients (unilateral cleft), the altered area extended to the contralateral, noncleft side of the upper lip or onto the philtrum. CONCLUSIONS: In contrast to the literature, sensation in the upper lip of many patients with cleft lip is not normal. Loss in sensation is exhibited most commonly and limited largely to the skin overlying tissues traumatized during reconstructive surgery. PMID- 15748111 TI - Bilateral asymmetry in Chinese families with cleft lip with or without cleft palate. AB - OBJECTIVE: To determine if Chinese individuals with nonsyndromic cleft lip with or without cleft palate (CL/P) display more bilateral asymmetry than do their unaffected relatives. DESIGN/SUBJECTS: A case-control study of 313 individuals with CL/P from Shanghai, China, with 201 unaffected relatives as controls. METHODS: Size-adjusted asymmetry scores were defined by data on middle-finger length, palm length, palpebral fissure width, and ear length. Case-control comparisons used a multivariate repeated measures analysis of variance, paired t tests, and the Wilcoxon signed rank test. RESULTS: The ear-length measure showed a significant increase in fluctuating asymmetry (FA) in individuals with CL/P compared with their unaffected relatives, which was most pronounced in the female cleft lip and palate subgroup (p = .04). No other measures showed any increase in FA. CONCLUSION: Evidence was found for increased FA, as measured by overall ear length, in Chinese individuals with nonsyndromic CL/P, compared with their unaffected family members. The use of bilateral measurements other than dermatoglyphics may prove to be a valuable means of assessing overall developmental stability in individuals with developmental malformations and in their families. PMID- 15748112 TI - Morphology of the auditory tube and palatal muscles in a case of bilateral cleft palate. AB - OBJECTIVE: There is an increased incidence of otitis media in children with cleft palate, which may be related to the pathology of the auditory tube and palatal muscles. In the present study, the head of a human on term born fetus with bilateral palatal cleft was serially sectioned and the anatomy of the auditory tube and palatal muscles were studied by computer-aided three-dimensional reconstruction. RESULTS: The results showed a nearly horizontal course of the auditory tube. The tensor veli palatini muscle had a bony attachment on either side. The levator veli palatini muscle also showed an abnormal course. CONCLUSIONS: This abnormal course may result in obstruction of the auditory tube during contraction. These pathological findings may explain the higher frequency of otitis media in children with cleft palate. PMID- 15748113 TI - Perceptual evaluation of hypernasality compared to HONC measures: the role of experience. AB - OBJECTIVES: Nasality ratings from experienced and inexperienced listeners were compared to accelerometric measures using the Horii Oral Nasal Coupling (HONC) Index to determine if one group's ratings are more closely related to the HONC Index measures. The reliability of listener ratings was studied to determine if experienced listeners had better reliability than inexperienced listeners. The influence of phonetic content was analyzed to learn if ratings of sentences with stop consonants yielded higher correlations with HONC scores than sentences containing glides. DESIGN: Experienced and inexperienced listeners' ratings of hypernasality were correlated to the HONC measures for two nonnasal sentence productions. Analysis of variance compared experienced versus inexperienced listener ratings. PARTICIPANTS: Ten listeners who were speech-language pathologists with at least 10 years of experience in assessing hypernasality and 10 listeners who were graduate students in communicative disorders with less than 1 year of experience. Speech samples were acquired from 13 children with varying degrees of hypernasality and 5 children with normal nasality. RESULTS: Correlations between ratings of hypernasality and HONC scores were .60 for the experienced group and .52 for the inexperienced group. In general, the experienced listeners rated the hypernasality of the speakers as less severe. Both groups had similar intrajudge reliability. Hypernasality ratings were not influenced by sentence context. CONCLUSIONS: Hypernasality can be rated in a reliable fashion regardless of listener experience. The correlations between the objective measure of nasalization (HONC) and the perceptual ratings were not as high as expected. Factors contributing to obtaining only moderate correlations will be discussed. PMID- 15748114 TI - Cleft palate and Beckwith-Wiedemann syndrome. AB - OBJECTIVE: Patients with Beckwith-Wiedemann syndrome suffer numerous anomalies, which vary somewhat from case to case. Cleft palate in combination with this syndrome has rarely been reported in the literature. Through two cases, this report examines the staging of the surgical repairs and the role of macroglossia in cleft palate and the consequences of the scarred palate on mandibular development. RESULTS: Of four patients with Beckwith-Wiedemann syndrome, only two had a cleft palate. The timing of the repair in these two children was different. Speech development was satisfactory in the first case but mediocre in the second. This result seemed to be related to a poor social environment. Mandibular prognathism persisted in both cases. CONCLUSION: The treatment of patients with cleft palate and Beckwith-Wiedemann syndrome remains complex. It is preferable not to operate on a cleft palate before performing a tongue reduction plasty, but rather to combine these two surgical interventions. This would reduce the risks of anesthesia and enable the palate to heal more efficiently. Surgical treatment should be performed after the age of 6 months and before problems in speech development occur. An orthognathic surgery at adolescence could be performed if prognathism persists. While the origin of the cleft palate is still being discussed, we cannot claim that macroglossia is related to the development of cleft palate, nor that the scarred palate has an impact on the mandibular development. PMID- 15748115 TI - Pacifier sucking in a child with cleft palate: occlusal consequences. AB - OBJECTIVE: The case of a child with a repaired cleft palate and a pacifier habit is presented. The influence of both the cleft and the habit on the malocclusion is discussed. The malocclusion disappeared as the habit ceased. PMID- 15748119 TI - Outcome measures measure outcomes, not effects of intervention. PMID- 15748120 TI - Do early quadriceps exercises affect the outcome of ACL reconstruction? A randomised controlled trial. AB - A prospective, blinded, randomised controlled trial investigated the effectiveness of quadriceps exercises following anterior cruciate ligament reconstruction. A treatment group (Quadriceps exercise group) performed straight leg raises and isometric quadriceps contractions throughout the first two postoperative weeks, and a second group (No quadriceps exercise group) did not. A battery of outcome measures assessed subjects postoperatively at day one, two weeks, and one, three and six months. A total of 103 patients (Quadriceps exercise n = 48, No quadriceps exercise n = 55) commenced the study with 91 subjects available at final follow up (Quadriceps exercise n = 47, No quadriceps exercise n = 44). Performance of quadriceps exercises significantly improved a number of knee flexion and extension range of motion measurements (p = 0.01 to 0.04). No significant differences were found between the two groups at any postoperative period for quadriceps lag (p = 0.36), functional hop testing (p = 0.49 to 0.51), isokinetic quadriceps strength (p = 0.70 to 0.72), the majority of numerical analogue scores (p = 0.1 to 0.94) and Cincinnati scores (p = 0.10 to 0.84). Subjects performing quadriceps exercises reported significantly higher pain scores with exercise on the first postoperative day (p = 0.02). At six months postoperatively, the Quadriceps exercise subjects reported significantly more favourable Cincinnati scores for symptoms (p = 0.005) and problems with sport (p = 0.05). While average knee laxity was not significantly different between treatment groups over time (p = 0.27 to 0.94), quadriceps exercise performance was associated with a significantly lower incidence of abnormal knee laxity. Isometric quadriceps exercises and straight leg raises can be safely prescribed during the first two postoperative weeks and confer advantages for faster recovery of knee range of motion and stability. It remains to be proven whether the magnitude of differences between groups is clinically significant. PMID- 15748121 TI - Movement detection at the ankle following stroke is poor. AB - This study assessed whether sense of movement is impaired at the ankle in persons post-stroke who are able to walk independently. Eleven chronic post-stroke subjects (> 4 months post stroke) who were ambulatory with or without walking aids and living within the community, and 10 healthy age-matched control subjects volunteered to participate. Proprioceptive acuity at the ankle, measured by sense of movement, was tested at three velocities, 0.1, 0.5, and 2.5 deg/sec, in random order. In addition, ankle range of motion and the distance that subjects walked in 6 minutes were assessed. Stroke subjects were significantly poorer (p < 0.001) at detecting movement at the affected ankle compared with either the unaffected ankle or with the control group at each of the velocities tested. Six out of 11 stroke subjects demonstrated significant impairment in movement detection compared to controls. The usual primary impairments following stroke are loss of strength and loss of co-ordination. However, reduced proprioceptive acuity at the affected ankle may also contribute to a person's ability to position and load the foot during walking. This could explain the moderate relationship found between proprioceptive acuity and walking endurance in persons following stroke (Spearman's rho = 0.63 to 0.77). PMID- 15748122 TI - Effects of manual hyperinflation and suctioning in respiratory mechanics in mechanically ventilated patients with ventilator-associated pneumonia. AB - Ventilator-associated pneumonia results from bacterial colonisation of the aerodigestive tract or aspiration of contaminated secretions into the lower airways. As a consequence of infection of the lung parenchyma and alveolitis, accumulation of inflammatory exudates and infiltration of airway mucosa can lead to unfavourable respiratory mechanics in ventilator-associated pneumonia. Tracheal suction is often employed by nursing staff in the management of mechanically ventilated patients with ventilator-associated pneumonia but this technique has the potential to increase respiratory resistance. Manual hyperinflation is used by physiotherapists to improve lung volume and mobilise secretions and has been shown to increase lung compliance. The effect of manual hyperinflation on airway resistance has not been studied. This study aims to demonstrate an additional mechanical benefit to the respiratory system when manual hyperinflation and suction techniques are combined, by comparing the application of manual hyperinflation and suction with suction alone on static lung compliance (C(L)) and inspiratory resistance (R(AW)) in mechanically ventilated patients with ventilator-associated pneumonia. Fifteen adult patients with ventilator-associated pneumonia were recruited and acted as their own controls. Manual hyperinflation followed by suction (manual hyperinflation plus suction) and suction alone were applied consecutively, in random order, on two occasions, four hours apart. Respiratory variables, C(L) and R(AW), were measured five times and the averaged value documented. Data were recorded before, immediately after, and 30 minutes after each intervention protocol. C(L) increased by 22% and R(AW) decreased by 21%, up to 30 minutes after manual hyperinflation plus suction, but not after suction alone. This study suggests that manual hyperinflation in conjunction with suction induces beneficial changes in respiratory mechanics in mechanically ventilated patients with ventilator associated pneumonia. PMID- 15748123 TI - Physiotherapy management of low back pain: does practice match the Dutch guidelines? AB - The purpose of this study is to explore adherence by Dutch physiotherapists to the physiotherapists' guideline for non-specific low back pain. For this study data from the National Information Service for Allied Health Care were used. This is a registration network that continuously collects information about physiotherapy patients and their treatment episodes. Within this network, adherence to the low back pain guideline was assessed by three criteria based on the guideline. These criteria concerned the number of sessions, the treatment goals, and the interventions. Data from patients with 'non-specific low back pain' as the reason for referral and a completed treatment episode were selected (n = 1254); 90 therapists in 40 practices treated these patients. The criterion concerning the number of sessions applied only for patients with acute complaints and was met in 17% of these patients. In about half of the patients the criterion for the treatment goals as well as the criterion relating to the interventions was met. Treatment goals are aimed mainly at improving mobility functions and changing body position. In more than three-quarters of the treatment episodes manual interventions (massage or manual manipulation) and exercise therapy were used frequently. As considerable variation in guideline adherence was shown to exist among therapists, there is clearly room for improvement in the quality of the care. PMID- 15748124 TI - Do changes within a manual therapy treatment session predict between-session changes for patients with cervical spine pain? AB - Physiotherapists often use within-session changes to provide a guide for refining treatment application. This study tested the validity of within-session changes as predictors of between-session changes for patients with neck pain receiving manual therapy treatment. A total of 70 pairs of treatments from 29 patients with sub-acute non-specific neck pain receiving manual therapy were assessed to determine the relationship between within-session and between-session changes in range of motion (ROM), pain intensity, and centralisation. Measurements were taken of ROM of the more limited direction on each axis of flexion, extension, lateral-flexion and rotation, and pain (intensity and location) before and after treatment. The same measurements were repeated before the following treatment. Regression analysis demonstrated that within-session change accounted for 26% to 48% of the variability in between-session change for ROM and six per cent for pain intensity. The proportion of the within-session change for ROM maintained between sessions ranged from 42% to 63% (95% CI 25% to 88%). The odds ratios for within-session improved/not improved categorisation to predict between-session category for ROM ranged from 2.5 (95% CI 0.6 to 4.3) to 21.3 (95% CI 10.1 to 96.1), for pain intensity 4.5 (95% CI 1.2 to 14.4) and for pain centralisation 9.2 (95% CI 2.2 to 38.7) indicating greater likelihood of between-session improvement after within-session improvement. The between-session results for most patients (71% to 83%) could be classified correctly by their within-session category. The results support the use of within-session changes in ROM, centralisation, and possibly pain intensity as predictors of between-session changes for musculoskeletal disorders of the cervical spine. PMID- 15748125 TI - Widespread brain activity during an abdominal task markedly reduced after pain physiology education: fMRI evaluation of a single patient with chronic low back pain. AB - The way people with chronic low back pain think about pain can affect the way they move. This case report concerns a patient with chronic disabling low back pain who underwent functional magnetic resonance imaging scans during performance of a voluntary trunk muscle task under three conditions: directly after training in the task and, after one week of practice, before and after a 2.5 hour pain physiology education session. Before education there was widespread brain activity during performance of the task, including activity in cortical regions known to be involved in pain, although the task was not painful. After education widespread activity was absent so that there was no brain activation outside of the primary somatosensory cortex. The results suggest that pain physiology education markedly altered brain activity during performance of the task. The data offer a possible mechanism for difficulty in acquisition of trunk muscle training in people with pain and suggest that the change in activity associated with education may reflect reduced threat value of the task. PMID- 15748126 TI - The shortage of kidneys for transplantation in Australia. PMID- 15748127 TI - Helicobacter pylori infection in Indigenous Australians: a serious health issue? AB - If Indigenous Australians are at high risk of H. pylori infection, the associated risks of peptic ulceration and gastric cancer may make screening and treatment necessary. PMID- 15748128 TI - Immunisation: reducing health inequality for Indigenous Australians. AB - Vaccination programs can act as a paradigm for effective health programs in Indigenous people. PMID- 15748129 TI - Prevalence of Helicobacter pylori in Indigenous Western Australians: comparison between urban and remote rural populations. AB - OBJECTIVES: To determine and compare the prevalence of Helicobacter pylori in an urban and a remote rural Western Australian Indigenous community. DESIGN: Cross sectional study of Helicobacter pylori status determined by urea breath tests between mid-January 2003 and the end of June 2004. PARTICIPANTS: 520 self selected fasting participants, comprising 270 members of the Martu community at Jigalong, Punmu and Parnngurr in the East Pilbara region (129 men, 141 women; age range, 2-90 years) and 250 people from the Perth Indigenous community (96 men, 154 women; age range, 3-75 years. RESULTS: The overall prevalence of H. pylori was 76%, but the prevalence in the remote rural community was 91%, compared with 60% in the urban community. The odds of having H. pylori were six times greater for rural than for urban participants (odds ratio [OR], 6.34; 95% CI, 3.89 10.33). Further, the overall odds of H. pylori infection in males (rural and urban combined) were greater than for females (OR, 1.61; 95% CI, 1.02-2.54). In both communities, the prevalence of infection remained relatively constant after the age of 10. CONCLUSIONS: The prevalence of H. pylori in the two Indigenous communities was two to three times higher than that in the non-Indigenous Australian population and higher than that shown in previous studies in Indigenous Australians. PMID- 15748130 TI - Effect of a consultation teaching behaviour modification on sleep performance in infants: a randomised controlled trial. AB - OBJECTIVE: To evaluate the effect of a behaviour modification program, taught to parents in a single visit to a trained nurse, in improving sleep performance in newborn infants, Australia. DESIGN: Randomised controlled trial. SETTING AND PARTICIPANTS: 268 families with normal newborn infants in the community, recruited between October 1996 and March 1997 from birth notices published in a South Australian daily newspaper. INTERVENTION: A 45-minute consultation with a nurse 2-3 weeks after the birth, including a tutorial discussion on normal sleep patterns in newborn infants, supported by retained written material and, for infants with weight gain < 30 g daily, referral to their usual postnatal care provider. MAIN OUTCOME MEASURES: Hours of daytime sleep (0600-1800), night sleep (1800-0600) and total sleep per 24 h; and number of daily records with total sleep >/= 15 h per 24 h, assessed by 7-day sleep diary at ages 6 and 12 weeks. RESULTS: 268 families returned at least one sleep diary (137/171 intervention, 131/175 control), recording 3273 days. Two intervention infants were referred for low weight gain. Total sleep time was 15 h or more per 24 h on 62% of recorded days in the intervention group, compared with 36% in the control group (P < 0.001). At 6 weeks of age, intervention infants slept a mean 1.3 h per day more than control infants (95% CI, 0.95-1.65), comprising a mean 0.5 h more night sleep (95% CI, 0.32-0.69) and 0.8 h more daytime sleep (95% CI, 0.56-1.07). At 12 weeks, intervention infants slept a mean 1.2 h per day more (95% CI, 0.94-2.14), comprising 0.64 h more night sleep (95% CI, 0.19-0.89) and 0.58 h more daytime sleep (95% CI, 0.39-1.03). There was no significant difference in crying time between the groups. CONCLUSIONS: A single consultation supported by written material in the first 3 weeks of a child's life improves sleep performance at 6 weeks of age. This improvement is maintained at 3 months. PMID- 15748131 TI - Impact of smoking, diabetes and hypertension on survival time in the elderly: the Dubbo Study. AB - OBJECTIVE: To study the impact of various risk factors on survival time in a cohort of elderly Australians. DESIGN, SETTING AND PARTICIPANTS: A longitudinal, prospective cohort study conducted in Dubbo, NSW. Participants were men and women aged 60 years or over living in the community, first assessed in 1988-1989 and followed for 15 years. MAIN OUTCOME MEASURES: Mortality rates; risk factors; survival times. RESULTS: There were 668 deaths in 1233 men (54%) and 625 deaths in 1572 women (40%). Coronary heart disease was the major cause of death, rates being higher in men than women until age group 80+ years; stroke death rates were similar in both sexes; cancer and respiratory death rates were higher in men than women across all ages. In a proportional hazards model, the independent predictors of mortality were cigarette smoking, diabetes, very high blood pressure (BP), impaired peak expiratory flow (PEF), physical disability, and zero intake of alcohol. Over 15 years, the average reductions in survival time associated with various risk factors, in men and women respectively, were smoking, 22 and 15 months; diabetes, 18 and 18 months; very high BP, 16 and 9 months; impaired PEF, 14 and 17 months; physical disability, 16 and 12 months; zero alcohol intake, 9 and 5 months. Combinations of selected risk factors were associated with a multiplier effect. CONCLUSION: The reduction in survival time in elderly citizens demonstrated in the presence of smoking, diabetes and hypertension highlights a potential benefit to healthy ageing to be gained from prevention and intervention. PMID- 15748133 TI - Achieving equal standards in medical student education: is a national exit examination the answer? AB - Although it is commonly assumed that the quality of medical school education in Australia is uniformly high, there is no national process for assessing its outcomes. There is substantial variability in the content of medical school curricula, and the process of curriculum change is becoming more challenging because of intense competition for time and space in the course. A national exit examination could provide a uniform standard of assessment for all medical school graduates in Australia, as well as foreign graduates applying to work in Australia. Such an examination could assess medical school outcomes, monitor the effects of curriculum change, and provide a benchmark for new medical schools that would help medical curricula evolve to better meet society's needs. PMID- 15748134 TI - A new focus of Rickettsia honei spotted fever in South Australia. AB - We recently diagnosed rickettsial spotted fever in four patients from the south eastern coastal region of South Australia near Adelaide, an area not known to be endemic for this infection. All infections were acquired within the geographic range of Aponomma hydrosauri, the tick vector of Rickettsia honei. Infection by R. honei was confirmed in two patients. This extension of the known geographic range of R. honei infection may be explained, in part, by alterations in host parasite ecology. PMID- 15748132 TI - Outcome of overseas commercial kidney transplantation: an Australian perspective. AB - Lack of donors has led to a worldwide increase in commercial kidney transplantation programs where recipients acquire kidneys either from executed prisoners or live non-related donors. Commercial transplantation is prohibited by legislation in Australia. Our centres have had 16 patients who have travelled overseas to receive a commercial kidney transplant; five have subsequently died. As has been found previously, patients who received commercial transplants were more likely to develop infections such as HIV, hepatitis B virus, cytomegalovirus and fungal infections. Previous reports have found that patient and graft survival were comparable to local results, whereas we found that patient and graft survival were worse than transplantation within Australia. Patients considering the option of overseas commercial donation should be advised that heightened risks to life and graft survival exist. PMID- 15748135 TI - Acute interstitial nephritis secondary to esomeprazole. PMID- 15748136 TI - Transdermal progesterone creams for postmenopausal women: more hype than hope? AB - Various claims have been made about the benefits of transdermal progesterone creams for relieving symptoms of menopause. Peer-reviewed articles have reported that the creams can raise plasma progesterone levels slightly, but have no effect on vasomotor, psychosexual or mood symptoms, bone metabolism or plasma lipid levels. Currently available progesterone creams can not be recommended for treatment of symptoms associated with menopause. PMID- 15748137 TI - 5. Constipation and toileting issues in children. AB - Constipation is common in children, with prevalences ranging from 0.3% up to 28%. In most children, constipation is functional (ie, without objective evidence of a pathological condition). Painful defecation has been proposed as the primary precipitant of functional faecal retention in early childhood. Faecal soiling is often secondary to constipation, and may occur during spontaneous relaxation of the sphincters precipitated by rectal distension. Management in general practice combines behavioural modification techniques with prolonged courses of laxatives. Treatment is usually successful, but may take up to 6-12 months. Significant numbers of children with initially good responses to therapy for constipation relapse in the long term. Long-term relapse is more frequent in children under 4 years at onset of symptoms and in whom there is a history of faecal soiling associated with constipation. PMID- 15748138 TI - Impact of a formal removal policy for central venous catheters on duration of catheterisation. PMID- 15748139 TI - Severe childhood pneumonitis caused by the Queensland strain of community acquired methicillin-resistant Staphylococcus aureus. PMID- 15748140 TI - Inhalation-device polypharmacy in asthma. PMID- 15748141 TI - Smoking and pregnancy. PMID- 15748142 TI - Trends in the use of hospital beds by older people in Australia: 1993-2002. PMID- 15748143 TI - Whistleblowing in the Australian public hospital system. PMID- 15748144 TI - The direct thrombin inhibitor melagatran/ximelagatran. PMID- 15748145 TI - Prescription Shoppers Line. PMID- 15748146 TI - Australasian Association of Doctors' Health Advisory Services. PMID- 15748147 TI - Increasing the culture efficiency of hybridoma cells by the use of integrated metabolic control of glucose and glutamine at low levels. AB - The metabolism of HAb18 hybridoma cells was shifted to decrease metabolite accumulation and to improve culture efficiency by integrated metabolic control of glucose and glutamine at low levels. When glucose and glutamine levels were decreased to 0.5 and 0.3 mM respectively, lactate and ammonia production were reduced by 62.6 and 74% respectively, glucose-to-cell yield was increased from 0.23x10(9) to 0.66x10(9) cells.mmol-1, and glutamine-to-cell yield from 0.18x10(9) to 1.95x10(9) cells.mmol-1. Compared with high-level glucose and glutamine fed-batch cultures, low-level glucose and glutamine led to higher cell density (1.0x10(7) versus 0.3x10(7) cells.ml-1), longer culture span (14 as opposed to 8 days) and higher antibody yield (250 as against 150 mg.l-1). These results indicate that hybridoma culture efficiency would be increased by the integrated control of glucose and glutamine at 0.5 and 0.3 mM respectively. In contrast with previously reported glucose-and/or-glutamine-level-controlled fed batch cultures, we demonstrated an efficient strategy of nutrient level selection and amino acid feeding. More importantly, our accurately and well-distributed Equable Feeding Control System opens a new avenue for reducing metabolites to low levels by controlling nutrients at low levels. PMID- 15748148 TI - Heterodimerization of opioid receptor-like 1 and mu-opioid receptors impairs the potency of micro receptor agonist. AB - Nociceptin activation of ORL1 (opioid receptor-like 1 receptor) has been shown to antagonize mu receptor-mediated analgesia at the supraspinal level. ORL1 and mu opioid receptor (muR) are co-expressed in several subpopulations of CNS neurons involved in regulating pain transmission. The amino acid sequence of ORL1 also shares a high degree of homology with that of mu receptor. Thus, it is hypothesized that ORL1 and muR interact to form the heterodimer and that ORL1/muR heterodimerization may be one molecular basis for ORL1-mediated antiopioid effects in the brain. To test this hypothesis, myc-tagged ORL1 and HA-tagged muR are co-expressed in human embryonic kidney (HEK) 293 cells. Co immunoprecipitation experiments demonstrate that ORL1 dimerizes with muR and that intracellular C-terminal tails of ORL1 and muR are required for the formation of ORL1/muR heterodimer. Second messenger assays further indicate that formation of ORL1/muR heterodimer selectively induces cross-desensitization of muR and impairs the potency by which [D-Ala(2),N-methyl-Phe(4),Gly-ol(5)]enkephalin (DAMGO) inhibits adenylate cyclase and stimulates p42/p44 mitogen-activated protein kinase (MAPK) phosphorylation. These results provide the evidence that ORL1/muR heterodimerization and the resulting impairment of mu receptor-activated signaling pathways may contribute to ORL1-mediated antiopioid effects in the brain. PMID- 15748149 TI - Resistance to fipronil in Drosophila simulans: influence of two point mutations in the RDL GABA receptor subunit. AB - The Eyguieres 42 strain of Drosophila simulans, obtained by laboratory selection, displayed approximately 20,000-fold resistance to the insecticide fipronil. Molecular cloning of the cDNA encoding the RDL GABA receptor subunit of this strain revealed the presence of two mutations: the Rdl mutation (A301G) and an additional mutation in the third transmembrane domain (T350M). In order to assess the individual and combined roles of the two mutations in fipronil resistance, the functional properties of wild-type, A301G, T350M and A301G/T350M homomultimeric RDL receptors were compared by expression in Xenopus oocytes. In wild-type receptors, the inhibition of GABA (EC(30))-induced currents by fipronil and picrotoxin was enhanced by repeated GABA applications. The A301G mutation nearly abolished this effect, decreased the sensitivity to fipronil and picrotoxin and increased the reversibility of inhibition. The T350M mutation also reduced the sensitivity to both antagonists. Of the four receptor variants tested, the double mutant showed the highest resistance to fipronil, following repeated GABA applications. In conclusion, the present study emphasizes new aspects of the pharmacological alterations induced by the Rdl mutation and shows that resistance to GABA receptor-directed insecticides may implicate a mutation distinct from Rdl. PMID- 15748150 TI - Phosphorylation state of postsynaptic density proteins. AB - The postsynaptic density (PSD) is an electron-dense structure located at the synaptic contacts between neurons. Its considerable complexity includes cytoskeletal and scaffold proteins, receptors, ion channels and signaling molecules, in line with the role of PSDs in signal transduction and processing. The phosphorylation state of components of the PSD is central to synaptic transmission and is known to play a role in synaptic plasticity, learning and memory. The presence of a range of kinases and phosphatases in the PSD defines potential key players in this context. However, the substrates that these enzymes target have not been fully identified to date. We analyzed the protein composition of purified PSD samples from adult mouse brains by strong cation exchange chromatography fractionation of a tryptic digest followed by nano reverse phase liquid chromatography coupled with electrospray ionization quadrupole time of flight tandem mass spectrometry. This led to the identification of 244 proteins. To gain an insight into the phosphoproteome of the PSD we then purified phosphorylated tryptic peptides by immobilized metal ion affinity chromatography. This approach for the specific enrichment of phosphopeptides resulted in the identification of 42 phosphoproteins in the PSD preparation, 39 of which are known PSD components. Here we present a total of 83 in vivo phosphorylation sites. PMID- 15748151 TI - In vitro 1H NMR spectroscopy shows an increase in N-acetylaspartylglutamate and glutamine content in the hippocampus of amygdaloid-kindled rats. AB - We examined energy metabolism and amino acid content in the hippocampus of amygdaloid-kindled rats using (1)H NMR spectroscopy. Three weeks after the last stage 5 seizure, kindled rats were killed by microwave irradiation. The hippocampus was dissected out and subjected to MeOH/CHCl(3) extraction. All (1)H spectra were analyzed to quantify absolute concentrations using a non-linear least squares method, combined with a prior knowledge of chemical shifts. Saturation effects were compensated for by the T1 measurement of each component. Levels of energy metabolism-related compounds, phosphocreatine, creatine, glucose and succinate were the same in both kindled rats and sham controls. Lactate concentration had a tendency to increase, although this was not statistically significant. When compared with sham controls, levels of aspartate, glutamate, glycine and glutamine, as well as GABA and inositol, were increased in the ipsilateral but not the contralateral hippocampus. In contrast, levels of taurine, alanine and threonine were unchanged. Finally, N-acetylaspartylglutamate content was elevated, whereas N-acetyl-l-aspartate content was unaltered in the ipsilateral hippocampus of kindled animals. Our results suggest that amygdala kindling may affects amino acid metabolism, but not energy metabolism. PMID- 15748152 TI - Pharmacological modulation of the endocannabinoid system in a viral model of multiple sclerosis. AB - Theiler's virus infection of the central nervous system (CNS) induces an immune mediated demyelinating disease in susceptible mouse strains and serves as a relevant infection model for human multiple sclerosis (MS). Cannabinoids have been shown to exert beneficial effects on animal models of MS and evidence suggests that the endocannabinoid system plays a role in the tonic control of spasticity. In this study we show that OMDM1 [(R)-N-oleoyl-(1'-hydroxybenzyl)-2' ethanolamine] and OMDM2 [(S)-N-oleoyl-(1'-hydroxybenzyl)-2'-ethanolamine], two selective inhibitors of the putative endocannabinoid transporter and hence of endocannabinoid inactivation, provide an effective therapy for Theiler murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD). Treatment of TMEV-infected mice with OMDM1 and OMDM2 enhanced anandamide levels in the spinal cord and ameliorated motor symptoms. This was associated with a down-regulation of inflammatory responses in the spinal cord. In addition we show that OMDM1 and OMDM2 down-regulate macrophage function by (i) decreasing the surface expression of major histocompatibility complex (MHC) class II molecules, (ii) inhibiting nitric oxide synthase-2 (NOS-2) expression and (iii) reducing the production of the pro-inflammatory cytokines interleukin-1beta (IL-1beta) and interleukin-12 (IL-12p40). Taken together, these results point to the manipulation of the endocannabinoid system as a possible strategy to develop future MS therapeutic drugs. PMID- 15748153 TI - Hypoxia sensitizes human malignant glioma cells towards CD95L-induced cell death. AB - Death ligands such as CD95 ligand (CD95L) have limited activity against glioma cells under normoxic conditions. Hypoxia is a critical aspect of the microenvironment of gliomas in vivo. We investigated the effect of co-exposure to acute hypoxia and CD95 ligand in three human malignant glioma cell lines with different susceptibility to CD95L under normoxic conditions. Hypoxia sensitized all three cell lines towards CD95L-induced cell death. Co-exposure resulted in apoptotic changes in the early phase, with gradual conversion to secondary necrosis with increasing length of hypoxia. The mitochondrial injury induced by hypoxia was enhanced by co-treatment, and caspase cleavage became prominent. Inhibition of the epidermal growth factor receptor (EGFR), although sensitizing glioma cells to CD95L under normoxia, protects glioma cells from hypoxia by reducing energy consumption. However, the opposing effects of EGFR signalling on death induced by CD95L or hypoxia were neutralized by co-exposure to hypoxia and CD95L. Furthermore, inhibition of protein synthesis by cycloheximide also reduced glucose consumption and conferred protection from hypoxia, but did not modulate CD95L-induced cell death under hypoxic conditions. These results suggest that death ligands may be useful to target hypoxic tumour cells resistant to conventional therapies or to complement strategies aiming at the induction of tumour hypoxia. PMID- 15748154 TI - Short- and long-term enzymatic regulation secondary to metabolic insult in the rat retina. AB - Changes in oxygen and/or glucose availability may result in altered levels of ATP production and amino acid levels, and alteration in lactic acid production. However, under certain metabolic insults, the retina demonstrates considerable resilience and maintains ATP production, and/or retinal function. We wanted to investigate whether this resilience would be reflected in alterations in the activity of key enzymes of retinal metabolism, or enzymes associated with amino acid production that may supply their carbon skeleton for energy production. Enzymatic assays were conducted to determine the activity of key retinal metabolic enzymes total ATPase and Na(+)/K(+)-ATPase, aspartate aminotransferase and lactate dehydrogenase. In vitro anoxia led to an increase in retinal lactate dehydrogenase activity and to a decrease in retinal aspartate aminotransferase activity, without significant changes in Na(+)/K(+)-ATPase activity. In vivo inhibition of glutamine synthetase resulted in a short-term significant decrease in retinal aspartate aminotransferase activity. An increase in retinal aspartate aminotransferase and lactate dehydrogenase activities was accompanied by altered levels of amino acids in neurons and glia after partial inhibition of glial metabolism, implying that short- and long-term up- and down-regulation of key metabolic enzymes occurs to supply carbon skeletons for retinal metabolism. ATPase activity does not appear to fluctuate under the metabolic stresses employed in our experimental procedures. PMID- 15748155 TI - Pure albumin is a potent trigger of calcium signalling and proliferation in microglia but not macrophages or astrocytes. AB - Microglial activation is implicated in the neurotoxicity of neurodegenerative diseases. Raised intracerebral levels of albumin are associated with the pathology of Alzheimer's disease, multiple sclerosis, and stroke where blood brain barrier damage is evident. We report here that treatment of primary cultured microglia and the N9 microglial cell line with pure albumin, or albumin in which fatty acids and immunoglobulins remain attached (fraction V), induced a rise in intracellular calcium. This rise in intracellular calcium was mediated via Src tyrosine kinase and phospholipase C. The albumin-induced calcium response was coupled to microglial proliferation, which was prevented by BAPTA, U73122 or PP2 but not mimicked by thapsigargin. In contrast, peritoneal macrophages were resistant to albumin- or fraction V-induced calcium responses and proliferation, whilst primary cultured astrocytes or the TSA-3 astrocyte cell line were responsive to fraction V albumin but not pure albumin. Furthermore, cerebellar granule neurones did not respond to albumin. These data suggest that albumin may play a role in microglial activation in pathological situations involving blood brain barrier impairment, and that the specific responses of microglia to albumin allow a distinction to be made between the signalling responses of microglia, blood-borne macrophages, astrocytes and neurones. PMID- 15748156 TI - Changes in phosphorylation of the NMDA receptor in the rat hippocampus induced by status epilepticus. AB - Systemic administration of pilocarpine preceded by lithium induces status epilepticus (SE) that results in neurodegeneration and may lead to the development of spontaneous recurrent seizures. We investigated the effect of Li/pilocarpine-induced SE on phosphorylation of the NMDA receptor in rat hippocampus. Phosphorylation of NR1 by PKC on Ser890 was decreased to 45% of control values immediately following 1 h of SE. During the first 3 h following the termination of SE, phosphorylation of Ser890 increased 4-fold before declining to control values by 24 h. Phosphorylation of NR1 by PKA was also depressed relative to controls immediately following SE and transiently increased above control values upon the termination of SE. SE was accompanied by a general increase in tyrosine phosphorylation of hippocampal proteins that lasted for several hours following the termination of seizures. Tyrosine phosphorylation of the NR2A and NR2B subunits of the NMDAR increased 3-4-fold over control values during SE, continued to increase during the first hour following SE and then declined to control levels by 24 h. SE resulted in the activation of Src and Pyk2 associated with the postsynaptic apparatus, suggesting a role for these enzymes in the SE-induced increase in tyrosine phosphorylation. Changes in phosphorylation of the NMDA receptor may play a role in the pathophysiological consequences of SE. PMID- 15748157 TI - Neuroprotective effects of atorvastatin against glutamate-induced excitotoxicity in primary cortical neurones. AB - Statins [3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors] exert cholesterol-independent pleiotropic effects that include anti-thrombotic, anti-inflammatory, and anti-oxidative properties. Here, we examined direct protective effects of atorvastatin on neurones in different cell damage models in vitro. Primary cortical neurones were pre-treated with atorvastatin and then exposed to (i) glutamate, (ii) oxygen-glucose deprivation or (iii) several apoptosis-inducing compounds. Atorvastatin significantly protected from glutamate induced excitotoxicity as evidenced by propidium iodide staining, nuclear morphology, release of lactate dehydrogenase, and mitochondrial tetrazolium metabolism, but not from oxygen-glucose deprivation or apoptotic cell death. This anti-excitototoxic effect was evident with 2-4 days pre-treatment but not with daily administration or shorter-term pre-treatment. The protective properties occurred independently of 3-hydroxy-3-methylglutaryl-CoA reductase inhibition because co-treatment with mevalonate or other isoprenoids did not reverse or attenuate neuroprotection. Atorvastatin attenuated the glutamate-induced increase of intracellular calcium, which was associated with a modulation of NMDA receptor function. Taken together, atorvastatin exerts specific anti-excitotoxic effects independent of 3-hydroxy-3-methylglutaryl-CoA reductase inhibition, which has potential therapeutic implications. PMID- 15748158 TI - Combined pharmacological, mutational and cell biology approaches indicate that p53-dependent caspase 3 activation triggered by cellular prion is dependent on its endocytosis. AB - We have previously established that cellular prion PrP(c) elicited p53-dependent caspase 3 activation in various transfected cells and primary cultured neurons. Although we showed that PrP(c) modulates p53 expression at both transcriptional and post-transcriptional levels, it remained unclear as to whether cellular prion signals at the membrane to trigger intracellular messages or if prion proapoptotic activity necessitated its translocation into the cytoplasm. Here, we compare the processing and cell death-related functions of PrP(c) with those of a mutated PrP(c) protein (N-3F4 MoPrP(c)) in which three basic N-terminal residues responsible for PrP(c) internalization had been mutated. As expected, N-3F4 MoPrP(c) remains exclusively located at the membrane, whereas PrP(c) partitions between membrane-associated and intracellular compartments, but both, proteins undergo constitutive and protein kinase C-regulated disintegrin-mediated proteolysis, leading to N1 fragment production. Unlike PrP(c), N-3F4 MoPrP(c) expression does not induce caspase 3 activation after stimulation by staurosporine and was inert on p53 expression and promoter transactivation in both human cells and TSM1 mouse neurons. Interestingly, PrP(c)-induced caspase 3 activation was closely linked to its endocytosis. This phenotype was enhanced by proteasomal inhibition and prevented by sucrose treatment. Accordingly, immunohistochemical analysis showed that protection towards degradation increased intracellular PrP(c)-like immunoreactivity, while sucrose treatments fully abolished PrP(c) intracellular expression and co-localization with transferrin. Altogether, we, establish here, using combined biochemical, mutational and cell biology approaches, that the caspase 3 activation associated with cellular prion is closely related to its ability to undergo endocytosis. This is, to our knowledge, the first direct description of an endocytosis-dependent PrP(c) associated function. PMID- 15748159 TI - D2 dopamine receptor activation of potassium channels is selectively decoupled by Galpha-specific GoLoco motif peptides. AB - The GoLoco motif is a short polypeptide sequence found in G-protein signaling regulators such as regulator of G-protein signaling proteins type 12 and 14 and activator of G-protein signaling protein type 3. A unique property of the GoLoco motifs from these three proteins is their preferential interaction with guanosine diphosphate (GDP)-bound Galpha(i1), Galpha(i3) and, sometimes, Galpha(i2) subunits over Galpha(o) subunits. This interaction prevents both spontaneous guanine nucleotide release and reassociation of Galpha(i)-GDP with Gbetagamma. We utilized this property of the GoLoco motif to examine dopamine (D2 and D3) and somatostatin receptor coupling to G-protein-regulated inwardly rectifying potassium (GIRK) channels in mouse AtT20 cells. GoLoco motif peptides had no effect on either basal channel activity or the initial responses to agonists, suggesting that the GoLoco motif cannot disrupt pre-formed G-protein heterotrimers. GoLoco motif peptides did, however, interfere with human D2((short)) receptor coupling to GIRK channels as demonstrated by the progressively diminished responses after repeated agonist application. This behavior is consistent with some form of compartmentalization of D2 receptors and GIRK channels such that Gbetagamma subunits, freed by local receptor activation and prevented from reforming a heterotrimeric complex, are not functionally constrained within the receptor-channel complex and thus are unable to exert a persistent activating effect. In contrast, GoLoco motif peptides had no effect on either D3 or somatostatin coupling to GIRK channels. Our results suggest that GoLoco motif-based peptides will be useful tools in examining the specificity of G-protein-coupled receptor-effector coupling. PMID- 15748160 TI - ATP inhibits the hypoxia response in type I cells of rat carotid bodies. AB - Summary During hypoxia, ATP was released from type I (glomus) cells in the carotid bodies. We studied the action of ATP on the intracellular Ca(2+) concentration ([Ca(2+)](i)) of type I cells dissociated from rat carotid bodies using a Ca(2+) imaging technique. ATP did not affect the resting [Ca(2+)](i) but strongly suppressed the hypoxia-induced [Ca(2+)](i) elevations in type I cells. The order of purinoreceptor agonist potency in inhibiting the hypoxia response was 2-methylthioATP > ATP > ADP >> alpha, beta-methylene ATP > UTP, implicating the involvement of P2Y(1) receptors. Simultaneous measurements of membrane potential and [Ca(2+)](i) show that ATP inhibited the hypoxia-induced Ca(2+) signal by reversing the hypoxia-triggered depolarization. However, ATP did not oppose the hypoxia-mediated inhibition of the oxygen-sensitive TASK-like K(+) background current. Neither the inhibition of the large-conductance Ca(2+) activated K(+) (maxi-K) channels nor the removal of extracellular Na(+) could affect the inhibitory action of ATP. Under normoxic condition, ATP caused hyperpolarization and increase in cell input resistance. These results suggest that the inhibitory action of ATP is mediated via the closure of background conductance(s) other than the TASK-like K(+), maxi-K or Na(+) channels. In summary, ATP exerts strong negative feedback regulation on hypoxia signaling in rat carotid type I cells. PMID- 15748161 TI - PKC site mutations reveal differential modulation by insulin of NMDA receptors containing NR2A or NR2B subunits. AB - Insulin modulates N-methyl-d-aspartate (NMDA) receptors in the CNS and potentiates currents of recombinant NMDA receptors in a subunit-specific manner in Xenopus oocytes. Previously we identified two sites in the NR2B C-terminus as targets for direct phosphorylation by C-type protein kinases (PKCs). Mutating these sites reduced insulin potentiation of currents by one half, reflecting the PKC-mediated portion of the NR2B insulin effect. The PKC-proline rich tyrosine kinase (Pyk2)-Src family kinase pathway may also mediate insulin potentiation. A dominant negative Pyk2 mutant significantly reduced insulin potentiation when co expressed with NR2B-containing receptors, suggesting that Pyk2 and downstream Src family tyrosine kinases are involved, along with PKCs, in insulin potentiation of NR2B. The NR2A C-terminus contains two residues homologous to the NR2B PKC targets. Mutating both these sites eliminated insulin potentiation of NR2A containing receptors, while co-expression of dominant negative Pyk2 had no effect. Together, these data indicate that PKCs alone mediate the NR2A insulin effect. When tested individually for importance in insulin potentiation, the two PKC sites showed an additive effect in potentiation of NR2A-containing receptors. Insulin modulation of NR2A-containing receptors is mediated solely by PKCs, whereas insulin modulation of NR2B-containing receptors is mediated by PKCs and tyrosine kinases (PTKs). PMID- 15748162 TI - Inhibition of microglial inflammation by the MLK inhibitor CEP-1347. AB - CEP-1347 is a potent inhibitor of the mixed lineage kinases (MLKs), a distinct family of mitogen-activated protein kinase kinase kinases (MAPKKK). It blocks the activation of the c-Jun/JNK apoptotic pathway in neurons exposed to various stressors and attenuates neurodegeneration in animal models of Parkinson's disease (PD). Microglial activation may involve kinase pathways controlled by MLKs and might contribute to the pathology of neurodegenerative diseases. Therefore, the possibility that CEP-1347 modulates the microglial inflammatory response [tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and monocyte chemotactic protein-1 (MCP-1)] was explored. Indeed, the MLK inhibitor CEP-1347 reduced cytokine production in primary cultures of human and murine microglia, and in monocyte/macrophage-derived cell lines, stimulated with various endotoxins or the plaque forming peptide Abeta1-40. Moreover, CEP-1347 inhibited brain TNF production induced by intracerebroventricular injection of lipopolysaccharide in mice. As expected from a MLK inhibitor, CEP-1347 acted upstream of p38 and c-Jun activation in microglia by dampening the activity of both pathways. These data imply MLKs as important, yet unrecognized, modulators of microglial inflammation, and demonstrate a novel anti-inflammatory potential of CEP-1347. PMID- 15748163 TI - Induction of galanin receptor-1 (GalR1) expression in external granule cell layer of post-natal mouse cerebellum. AB - Galanin is a modulator of fast transmission in adult brain and recent evidence suggests that it also acts as a trophic factor during neurogenesis and neural injury and repair. Previous studies in our laboratory have identified galanin mRNA in Purkinje cells of adult and developing rat (but not adult mouse) cerebellum; and galanin-binding sites in adult mouse (but not rat) cerebellum. The post-natal development of the cerebellum provides a unique and convenient model for the investigation of developmental processes and to learn more about putative cerebellar galanin systems, the current study examined the presence and distribution of galanin-like-immunoreactivity (- LI), [(125)I]-galanin binding sites and galanin receptor-1 (GalR1) mRNA in post-natal mouse cerebellum. Using autoradiography and in situ hybridization, [(125)I]-galanin binding sites and GalR1 mRNA were first detected on post-natal day 10 (P10) in the external germinal layer of all lobes and high levels were maintained until P14. Quantitative real-time PCR assays detected GalR1 mRNA in whole cerebellum across the post-natal period, with a strong induction and peak of expression at P10. Assessment of galanin levels in whole cerebellum by radioimmunoassay revealed relatively similar concentrations from P5 to P20 and in adult mice (80-170 pg/mg protein), with a significantly higher concentration (250 pg/mg, p < 0.01) detected at P3. These concentrations were some four- to six-fold lower than those in adult forebrain samples. Using immunohistochemistry, galanin-like-immuno reactivity was observed in prominent fibrous elements within the white matter tracts of the cerebellum at P3-5 and in more punctate elements in the internal granule cell layer and associated with the Purkinje cell layer at P12 and P20. Increased levels of GalR1 mRNA and galanin binding (attributed to GalR1) in the external granule cell layer at P10-12/(14) coincide with granule cell migration from the external to the inner granule cell layer and together with demonstrated effects of other neuropeptide-receptor systems suggest a role for GalR1 signalling in regulating this or related developmental processes. PMID- 15748164 TI - Signal transduction pathways implicated in neural recognition molecule L1 triggered neuroprotection and neuritogenesis. AB - The signal transduction pathways involved in adhesion molecule L1-triggered neuritogenesis and neuroprotection were investigated using the extracellular domain of mouse or human L1 in fusion with the Fc portion of human immunoglobulin G or L1 purified from mouse brain by affinity chromatography. Substrate L1 triggered neuritogenesis and neuroprotection depended on distinct but also overlapping signal transduction pathways and on the expression of L1 at the neuronal cell surface. PI3 kinase inhibitors, Src family kinase inhibitors as well as mitogen-activated protein kinase kinase inhibitors reduced both L1 triggered neuritogenesis and neuroprotection. In contrast, fibroblast growth factor receptor inhibitors, a protein kinase A inhibitor, and an inhibitor of cAMP-mediated signal transduction pathways, blocked neuritogenesis, but did not affect L1-triggered neuroprotection. Proteolytic cleavage of L1 or its interaction partners is necessary for both L1-mediated neuritogensis and neuroprotection. Furthermore, L1-triggered neuroprotection was found to be associated with increased phosphorylation of extracellular signal-regulated kinases 1/2, Akt and Bad, and inhibition of caspases. These observations suggest possibilities of differentially targeting signal transduction pathways for L1 dependent neuritogenesis and neuroprotection. PMID- 15748166 TI - Trophic factors attenuate nitric oxide mediated neuronal and axonal injury in vitro: roles and interactions of mitogen-activated protein kinase signalling pathways. AB - Inflammation in the central nervous system occurs in diseases such as multiple sclerosis and leads to axon dysfunction and destruction. Both in vitro and in vivo observations have suggested an important role for nitric oxide (NO) in mediating inflammatory axonopathy. The purposes of this study were to model inflammatory axonopathy in vitro and identify modulators of the process. Rat cortical neurones were cultured and exposed to an NO-donor plus potential protective factors. Cultures were then assessed for neuronal survival, axon survival and markers of intracellular signalling pathways. The NO-donor produced dose-dependent neuronal loss and a large degree of axon destruction. Oligodendrocyte conditioned medium (OCM) and insulin-like growth factor type-1 (IGF-1), but not glial cell line-derived neurotrophic factor (GDNF), improved survival of neurones exposed to NO donors. In addition p38 MAP kinase was activated by NO exposure and inhibition of p38 signalling led to neuronal and axonal survival effects. OCM and IGF-1 (but not GDNF) reduced p38 activation in NO-exposed cortical neurones. OCM, IGF-1 and GDNF improved axon survival in cultures exposed to NO, a process dependent on mitogen-activated protein kinase/extracellular signal-related kinase signalling. This study emphasizes that different mechanisms may underlie neuronal/axonal destructive processes, and suggests that trophic factors may modulate NO-mediated neurone/axon destruction via specific pathways. PMID- 15748165 TI - Protection by cholesterol-extracting cyclodextrins: a role for N-methyl-D aspartate receptor redistribution. AB - Cyclodextrins (CDs) are cyclic oligosaccharides composed of a lipophilic central cavity and a hydrophilic outer surface. Some CDs are capable of extracting cholesterol from cell membranes and can affect function of receptors and proteins localized in cholesterol-rich membrane domains. In this report, we demonstrate the neuroprotective activity of some CD derivatives against oxygen-glucose deprivation (OGD), N-methyl-D-aspartic acid (NMDA) and glutamate in cortical neuronal cultures. Although all CDs complexed with NMDA or glutamate, only beta-, methylated beta- and sulfated beta-CDs displayed neuroprotective activity and lowered cellular cholesterol. Only CDs that lowered cholesterol levels redistributed the NMDA receptor NR2B subunit, PSD-95 (postsynaptic density protein 95 kDa) and neuronal nitric oxide synthase (nNOS) from Triton X-100 insoluble membrane domains to soluble fractions. Cholesterol repletion counteracted the ability of methylated beta-CD to protect against NMDA toxicity, and reversed NR2B, PSD-95 and nNOS localization to Triton X-100 insoluble membrane fraction. Surprisingly, neuroprotective CDs had minimal effect on NMDA receptor-mediated increases in intracellular Ca(2+) concentration ([Ca(2+)](i)), but did suppress OGD-induced increases in [Ca(2+)](i). beta-CD, but not Mbeta-CD, also caused a slight block of NMDA-induced currents, suggesting a minor contribution to neuroprotection by direct action on NMDA receptors. Taken together, data suggest that cholesterol extraction from detergent-resistant microdomains affects NMDA receptor subunit distribution and signal propagation, resulting in neuroprotection of cortical neuronal cultures against ischemic and excitotoxic insults. Since cholesterol-rich membrane domains exist in neuronal postsynaptic densities, these results imply that synaptic NMDA receptor subpopulations underlie excitotoxicity, which can be targeted by CDs without affecting overall neuronal Ca(2+) levels. PMID- 15748167 TI - Specific induction of macrophage inflammatory protein 1-alpha in glial cells of Sandhoff disease model mice associated with accumulation of N-acetylhexosaminyl glycoconjugates. AB - Sandhoff disease is a lysosomal storage disease caused by simultaneous deficiencies of beta-hexosaminidase A (HexA; alphabeta) and B (HexB; betabeta), due to a primary defect of the beta-subunit gene (HEXB) associated with excessive accumulation of GM2 ganglioside (GM2) and oligosaccharides with N acetylhexosamine residues at their non-reducing termini, and with neurosomatic manifestations. To elucidate the neuroinflammatory mechanisms involved in its pathogenesis, we analyzed the expression of chemokines in Sandhoff disease model mice (SD mice) produced by disruption of the murine Hex beta-subunit gene allele (Hexb-/-). We demonstrated that chemokine macrophage inflammatory protein-1 alpha (MIP-1alpha) was induced in brain regions, including the cerebral cortex, brain stem and cerebellum, of SD mice from an early stage of the pathogenesis but not in other systemic organs. On the other hand, little changes in other chemokine mRNAs, including those of RANTES (regulated upon activation, normal T expressed and secreted), MCP-1 (monocyte chemotactic protein-1), SLC (secondary lymphoid tissue chemokine), fractalkine and SDF-1 (stromal derived factor-1), were detected. Significant up-regulation of MIP-1alpha mRNA and protein in the above mentioned brain regions was observed in parallel with the accumulation of natural substrates of HexA and HexB. Immunohistochemical analysis revealed that MIP 1alpha-immunoreactivity (IR) in the above-mentioned brain regions of SD mice was co-localized in Iba1-IR-positive microglial cells and partly in glial fibrillary acidic protein (GFAP)-IR-positive astrocytes, in which marked accumulation of N acetylglucosaminyl (GlcNAc)-oligosaccharides was observed from the presymptomatic stage of the disease. In contrast, little MIP-1alpha-IR was observed in neurons in which GM2 accumulated predominantly. These results suggest that specific induction of MIP-1alpha might coincide with the accumulation of GlcNAc oligosaccharides due to a HexB deficiency in resident microglia and astrocytes in the brains of SD mice causing their activation and acceleration of the progressive neurodegeneration in SD mice. PMID- 15748168 TI - Spinal p38beta isoform mediates tissue injury-induced hyperalgesia and spinal sensitization. AB - Antagonist studies show that spinal p38 mitogen-activated protein kinase plays a crucial role in spinal sensitization. However, there are two p38 isoforms found in spinal cord and the relative contribution of these two to hyperalgesia is not known. Here we demonstrate that the isoforms are distinctly expressed in spinal dorsal horn: p38alpha in neurons and p38beta in microglia. In lieu of isoform selective inhibitors, we examined the functional role of these two individual isoforms in nociception by using intrathecal isoform-specific antisense oligonucleotides to selectively block the expression of the respective isoform. In these rats, down-regulation of spinal p38beta, but not p38alpha, prevented nocifensive flinching evoked by intraplantar injection of formalin and hyperalgesia induced by activation of spinal neurokinin-1 receptors through intrathecal injection of substance P. Both intraplantar formalin and intrathecal substance P produced an increase in spinal p38 phosphorylation and this phosphorylation (activation) was prevented when spinal p38beta, but not p38alpha, was down-regulated. Thus, spinal p38beta, probably in microglia, plays a significant role in spinal nociceptive processing and represents a potential target for pain therapy. PMID- 15748169 TI - Inducers of oxidative stress block ciliary neurotrophic factor activation of Jak/STAT signaling in neurons. AB - Generation of reactive oxygen species (ROS) with the accumulation of oxidative damage has been implicated in neurodegenerative disease and in the degradation of nervous system function with age. Here we report that ROS inhibit the activity of ciliary neurotrophic factor (CNTF) in nerve cells. Treatment with hydrogen peroxide (H(2)O(2)) as a generator of ROS inhibited CNTF-mediated Jak/STAT signaling in all cultured nerve cells tested, including chick ciliary ganglion neurons, chick neural retina, HMN-1 motor neuron hybrid cells, and SH-SY5Y and BE(2)-C human neuroblastoma cells. H(2)O(2) treatment of non-neuronal cells, chick skeletal muscle and HepG2 hepatoma cells, did not inhibit Jak/STAT signaling. The H(2)O(2) block of CNTF activity was seen at concentrations as low as 0.1 mm and within 15 min, and was reversible upon removal of H(2)O(2) from the medium. Also, two other mediators of oxidative stress, nitric oxide and rotenone, inhibited CNTF signaling. Treatment of neurons with H(2)O(2) and rotenone also inhibited interferon-gamma-mediated activation of Jak/STAT1. Depleting the intracellular stores of reduced glutathione by treatment of BE(2)-C cells with nitrofurantoin inhibited CNTF activity, whereas addition of reduced glutathione protected cells from the effects of H(2)O(2). These results suggest that disruption of neurotrophic factor signaling by mediators of oxidative stress may contribute to the neuronal damage observed in neurodegenerative diseases and significantly affect the utility of CNTF-like factors as therapeutic agents in preventing nerve cell death. PMID- 15748170 TI - Impaired proteasome activity and accumulation of ubiquitinated substrates in a hereditary neuropathy model. AB - Accumulation of misfolded proteins and alterations in the ubiquitin-proteasome pathway are associated with various neurodegenerative conditions of the CNS and PNS. Aggregates containing ubiquitin and peripheral myelin protein 22 (PMP22) have been observed in the Trembler J mouse model of Charcot-Marie-Tooth disease type 1A demyelinating neuropathy. In these nerves, the turnover rate of the newly synthesized PMP22 is reduced, suggesting proteasome impairment. Here we show evidence of proteasome impairment in Trembler J neuropathy samples compared with wild-type, as measured by reduced degradation of substrate reporters. Proteasome impairment correlates with increased levels of polyubiquitinated proteins, including PMP22, and the recruitment of E1, 20S and 11S to aggresomes formed either spontaneously due to the Trembler J mutation or upon proteasome inhibition. Furthermore, myelin basic protein, an endogenous Schwann cell proteasome substrate, associates with PMP22 aggregates in affected nerves. Together, our data show that in neuropathy nerves, reduced proteasome activity is coupled with the accumulation of ubiquitinated substrates, and the recruitment of proteasomal pathway constituents to aggregates. These results provide novel insights into the mechanism by which altered degradation of Schwann cell proteins may contribute to the pathogenesis of certain PMP22 neuropathies. PMID- 15748171 TI - An adenosine A receptor agonist induces sleep by increasing GABA release in the tuberomammillary nucleus to inhibit histaminergic systems in rats. AB - The adenosine A(2A) receptor (A(2A)R) has been demonstrated to play a crucial role in the regulation of the sleep process. However, the molecular mechanism of the A(2A)R-mediated sleep remains to be elucidated. Here we used electroencephalogram and electromyogram recordings coupled with in vivo microdialysis to investigate the effects of an A(2A)R agonist, CGS21680, on sleep and on the release of histamine and GABA in the brain. In freely moving rats, CGS21680 applied to the subarachnoid space underlying the rostral basal forebrain significantly promoted sleep and inhibited histamine release in the frontal cortex. The histamine release was negatively correlated with the amount of non rapid eye movement sleep (r = - 0.652). In urethane-anesthetized rats, CGS21680 inhibited histamine release in both the frontal cortex and medial pre-optic area in a dose-dependent manner, and increased GABA release specifically in the histaminergic tuberomammillary nucleus but not in the frontal cortex. Moreover, the CGS21680-induced inhibition of histamine release was antagonized by perfusion of the tuberomammillary nucleus with a GABA(A) antagonist, picrotoxin. These results suggest that the A(2A)R agonist induced sleep by inhibiting the histaminergic system through increasing GABA release in the tuberomammillary nucleus. PMID- 15748172 TI - Purified Wnt-5a increases differentiation of midbrain dopaminergic cells and dishevelled phosphorylation. AB - The Wnt family of lipoproteins regulates several aspects of the development of the nervous system. Recently, we reported that Wnt-3a enhances the proliferation of midbrain dopaminergic precursors and that Wnt-5a promotes their differentiation into dopaminergic neurones. Here we report the purification of hemagglutinin-tagged Wnt-5a using a three-step purification method similar to that previously described for Wnt-3a. Haemagglutinin-tagged Wnt-5a was biologically active and induced the differentiation of immature primary midbrain precursors into tyrosine hydroxylase-positive dopaminergic neurones. Using a substantia nigra-derived dopaminergic cell line (SN4741), we found that Wnt-5a, unlike Wnt-3a, did not promote beta-catenin phosphorylation or stabilization. However, both Wnt-5a and Wnt-3a activated dishevelled, as assessed by a phosphorylation-dependent mobility shift. Moreover, the activity of Wnt-5a on dishevelled was blocked by pre-treatment with acyl protein thioesterase-1, indicating that palmitoylation of Wnt-5a is necessary for its function. Thus, our results suggest that Wnt-3a and Wnt-5a, respectively, activate canonical and non canonical Wnt signalling pathways in ventral midbrain dopaminergic cells. Furthermore, we identify dishevelled as a key player in transducing both Wnt canonical and non-canonical signals in dopaminergic cells. PMID- 15748175 TI - Module one: introduction to research ethics. AB - This module will introduce you to the ethical concepts underlying applied ethical decision-making in the area of research involving human participants. We will also learn what the issues are that people involved in research on research ethics are concerned with. Ethics without an understanding of historical and legal context makes arguably little sense. It is for this reason that this module will begin with a brief history of research ethics and ends with a brief overview of the relevant national and international guidelines pertaining to ethical issues in research involving human participants. PMID- 15748176 TI - Module two: informed consent. AB - The objective of this module is to familiarise you with the concept of informed consent, its ethical basis, its elements, and typical problems that are encountered even by the most well intentioned researchers when trying to achieve genuine informed consent. PMID- 15748177 TI - Module three: vulnerable/special participant populations. AB - ABSTRACT This module is designed to sensitise you to the special needs of participants who belong to populations that are more vulnerable than other participant populations. These populations typically include incompetent persons women who may or may not be pregnant, children, prisoners and refugees, impoverished people, and ethnic minorities. These and similar groups deserve special consideration for a number of important ethical and historical reasons, specifically those that surround the potential for exploitation, problems with informed consent, and concerns about respect for participant autonomy. This module introduces modus operandi that are based on national and international research guidelines for dealing with vulnerable/special participant populations, offering contextually-dependent advice and relevant ethical considerations/arguments for and against their involvement in scientific research endeavours. PMID- 15748178 TI - Module four: standards of care and clinical trials. AB - This module examines ethical debates about the level of care that should be provided to human research participants. Particular attention is placed on the question of what should be considered an ethically acceptable control arm. You will also learn what relevant international and domestic regulatory documents say about standards of care. PMID- 15748179 TI - Module five: implementation of ethics review. AB - The objective of this module is to inform you on issues of concern for Research Ethics Committee members and investigators during the review process. The many guidelines on research ethics, including those from the South African Department of Health and the World Health Organisation, will be referred to extensively to educate you on the requirements of Research Ethics Committees. The evolution of the review process in South Africa will be detailed. PMID- 15748180 TI - Module six: special issues. AB - The objective of this module is to cover ground that was not covered in-depth in any of the other modules, including: scientific misconduct, issues concerning the publication and ownership of research results (authorship guidelines - who is eligible to be considered an author, or contributor to a scientific paper etc.), special problems occurring in social science and epidemiological research, and the problems pertaining to conflicts of interest the various players in biomedical research activities could encounter. PMID- 15748181 TI - Otoplasty: a review of the surgical techniques. AB - Otoplasty surgery carries a high satisfaction rate for participants (96%). Surgeons are more critical of the results of otoplasty than the participants or parents (92% satisfied). There is no statistical difference in qualitative results with different surgical techniques when comparing the published data. Data on otoplasty surgery is numerous, but quantitative assessment of surgical outcome is rare. The authors propose the measurement of cephaloauricular distance to allow comparison between studies. PMID- 15748182 TI - Superior canal dehiscence: review of a new condition. AB - A new cause of sound and pressure induced vertigo, superior canal dehisence, is described. Auditory manifestations include hyperacusis to bone-conducted sounds and conductive hearing loss with normal acoustic reflexes. The diagnosis is reached by a directed history, documentation of upward and torsional nystagmus evoked by sound and pressure, and radiology. Acoustic reflexes and VEMP (vestibular evoked myogenic potentials) aid in the identification of patients with an apparent conductive loss with normal acoustic reflexes or have an asymptomatic dehiscense on radiology. Treatment involves avoidance of the precipitating stimuli. Surgical treatment, by resurfacing the dehiscence, is considered in patients with more severe symptoms. PMID- 15748183 TI - The relationship between self-reported difficulty with hearing in the worse ear and the better ear. AB - OBJECTIVES: To investigate the relationship of self-reported hearing difficulty with hearing in the worse ear and the better ear at 1 and 4 kHz. DESIGN: Receiver operating characteristic (ROC) curve analysis in community-residing population. SETTINGS: The Settsu City Health Center (Osaka, Japan). PARTICIPANTS: A total of 919 consecutive participants who were provided health check-ups. MAIN OUTCOMES MEASURES: Comparison is based on the measurement of the area under the curve (AUC) of ROC. RESULTS: At 1 kHz, the area under the curve (AUC) of the worse ear (0.625 +/- 0.021) was significantly larger than that of the better ear (0.570 +/- 0.021) (P < 0.01). At 4 kHz, the AUC of the worse ear (0.665 +/- 0.019) was significantly larger than that of the better ear (0.622 +/- 0.020) (P < 0.01). Excluding the subjects with noise exposure, tinnitus and ear disease, at both frequencies, the AUC of the worse ear was not significantly larger than that of the better ear at both 1 and 4 kHz. Hearing in the worse ear was significantly more related to self-reported hearing difficulty than in the better ear without excluding those with noise exposure, tinnitus and ear disease. CONCLUSIONS: ROC curve analysis would provide a method to help us judge the difference between the worse hearing ear and the better hearing ear on the subject of hearing loss. PMID- 15748184 TI - A prospective evaluation of the Essen titanium stapes prosthesis. AB - OBJECTIVES: This is the first clinical trial to evaluate the suitability of a new titanium stapes prosthesis, which we developed jointly with the Kurz Company (Dusslingen). DESIGN: In a prospective clinical study, patients with otosclerosis underwent stapes surgery using our new titanium stapes prosthesis during a period of 14 months. One year after surgery pre- and postoperative audiograms were performed and all patients participated in a structured interview. SETTING: Tertiary otorhinolaryngological university department PARTICIPANTS: Of 49 patients with otosclerosis, entering the study, two patients were excluded, because they were not located any more. MAIN OUTCOME MEASURES AND RESULTS: All patients experienced a hearing improvement of 21 dB on average except one patient. Sixty percentage of patients achieved a reduction of the air-bone gap to less than 10 dB and 31% of patients having their air-bone gap closed to within 20 dB--averaged across 0.5, 1.0, 2.0 and 4 kHz. The hearing level improved significantly in the air and in the bone conduction thresholds. CONCLUSIONS: This paper presents our first '1 year postimplantation' results of our titanium stapes prosthesis. We consider the development of this implant to be beneficial. The results confirm significant improvement in hearing status and tinnitus levels. PMID- 15748185 TI - The effect of mental alerting on gaze-evoked nystagmus. AB - OBJECTIVES: To determine whether mental alerting (MA) makes a significant difference to nystagmus recorded during gaze testing using electronystagmography (ENG). DESIGN: A prospective study, blinded interpretation of test results, alternate order of testing sequence. SETTING: Vestibular assessment clinic in a tertiary referral centre. PARTICIPANTS: A total of 80 patients were seen by the authors (R.L.H., K.F.L.M.) for vestibular assessment, between August 2002 and February 2004. Patients cognitively unable to perform MA tasks or with limited English were omitted. Patients all underwent gaze-testing using ENG. Each patient was tested twice, once without MA and once with an MA task, in an alternate test order. MAIN OUTCOME MEASURES: Slow-phase velocity values of recorded nystagmus, number of patients with clinically significant nystagmus (slow-phase velocity > or = 6 deg/s). RESULTS: A total of eight patients had significant gaze-evoked nystagmus when tested without MA and a total of six patients when tested with MA. Only four of these patients were common to both groups. When all nystagmus values (regardless of significance) were considered, no statistically significant differences between nystagmus measured with and without alerting were found (Wilcoxon signed rank test, P > 0.05). CONCLUSIONS: Mental alerting during gaze testing did not result in a significant difference in nystagmus and is just as likely to result in an exacerbation as a reduction in slow-phase velocity. In addition, MA is likely to result in a noisy (and therefore difficult to interpret) recording and increases the impact of the test on the patient. In view of these findings, we propose that MA is contraindicated during the gaze-testing component of the vestibular test battery. PMID- 15748186 TI - A blinded in-vitro study to compare the efficacy of five topical ear drops in clearing grommets blocked with thick middle ear effusion fluid. AB - OBJECTIVE: To compare the efficacy of 5% NaHCO3, 3% H2O2, Sofradex (dexamethasone sodium metasulphobenzoate 0.05%, framycetin sulphate 0.5%, gramicidin 0.005%), 0.33% acetic acid and 0.9% NaCl eardrops in clearing grommets blocked with harvested thick middle ear effusion fluid. STUDY DESIGN: A blinded in-vitro study. SETTING: District general hospital. PARTICIPANTS: A total of 473 grommets were blocked with freshly harvested unpooled thick middle ear effusion fluid obtained from 68 patients. MAIN OUTCOME MEASURES: Patency of the grommets before and 7 days after intervention was ascertained by tympanometry and close visual inspection. RESULTS: Instillation of eardrops leads to a statistically significant increase in the clearance of grommets as compared with not using any drops (chi2 = 14.3, d.f. = 5, P = 0.006). The numbers needed to treat were 2.8 for NaHCO3, 3.2 for 0.9% NaCl, 3.9 for 0.33% acetic Acid, 4.4 for Sofradex and 9.5 for H2O2 eardrops. Pair-wise comparison was only significant for comparison between 5% NaHCO3 and 3% H2O2 eardrops (Bonferroni corrected P = 0.01, odds ratio = 4.3, CI = 1.9-9.9). CONCLUSIONS: Use of eardrops leads to a clinically and statistically significant increase in the clearance of blocked grommets. Of the five drops used, 5% NaHCO3 was the most efficacious and 3% H2O2 the least efficacious. Limitations of this in-vitro study are recognized and a prospective in-vivo double blind randomized controlled trial is planned. PMID- 15748187 TI - Evidence for correlation of objective and subjective measures of nasal airflow in patients with common cold. AB - OBJECTIVES: To utilize posterior rhinomanometry and conductance, as units of measurement, to further investigate the relationship between subjective and objective measures of nasal airflow. DESIGN: A prospective, observational study. SETTING: Common Cold Research Centre. PARTICIPANTS: Sixty healthy volunteers from the staff and student population of Cardiff University with an upper respiratory tract infection. MAIN OUTCOMES MEASURES: To determine correlations between visual analogue scores (VAS) and posterior rhinomanometry for total, unilateral, high and low conductance groups. RESULTS: No correlation was found between total VAS and total conductance (r = 0.17, P = 0.10). A substantially significant correlation was found between unilateral VAS and unilateral conductance (rho = 0.50, P < 0.001). The unilateral VAS and conductance were highly correlated for the low total conductance group (rho = 0.61, P < 0.001). CONCLUSIONS: Posterior rhinomanometry allows actual measurement of the combined and unilateral conductance of nasal passages. The units of conductance, as opposed to resistance, allow totally obstructed nasal passages to be included in analysis. Visual analogue scores and conductance correlate strongly in unilateral measures for participants with a low total nasal conductance. Posterior rhinomanometry and units of conductance are recommended for future studies investigating the relationship between objective and subjective measures of nasal airflow. PMID- 15748188 TI - Adenoid size assessment: a comparison of palpation, nasendoscopy and mirror examination. AB - OBJECTIVES: To compare the digital, mirror and nasendoscopic assessment of adenoid size and posterior choanal obstruction in patients undergoing adenoidectomy. DESIGN: Prospective, blinded study. SETTING: Otorhinolaryngology department at a London teaching hospital. PARTICIPANTS: Twenty-eight consecutive patients undergoing adenoidectomy in conjunction with tonsillectomy or myringotomy under general anaesthesia, aged 17 months to 16 years. MAIN OUTCOME MEASURES: Adenoid size and postnasal space obstruction as assessed by digital examination, nasendoscopy and trans-oral mirror visualization. These examination methods were each compared with each other. RESULTS: Nasendoscopy and mirror examination correlated well (Spearman's R(S) = 0.71, P < 0.0001) but Passing and Bablock regression analysis demonstrated that mirror examination consistently underestimated the degree of choanal obstruction in comparison with nasendoscopy. There was no significant correlation between nasendoscopy and palpation (R(S) = 0.26, P = 0.17) and only a moderate correlation between mirror examination and palpation (R(S) = 0.46, P = 0.014). CONCLUSION: If nasendoscopy is considered the gold standard, then palpation is a poor measure of adenoid hypertrophy and mirror examination consistently underestimates choanal occlusion. PMID- 15748189 TI - Quality of life and voice following endoscopic resection or radiotherapy for early glottic cancer. AB - OBJECTIVES: To assess whether proposed voice and quality of life (QoL) outcome measures were likely to be acceptable to patients previously treated for early glottic cancer by either radiotherapy or endoscopic resection, as well as looking for differences in QoL and voice between treatments. DESIGN: Questionnaire-based cohort study. SETTING: Secondary care, three centres. PARTICIPANTS: All patients treated for T1a or in situ glottic carcinoma between 1997 and 2003. Fifty-three patients were identified; those who had undergone salvage surgery or radiotherapy were excluded. A proportion refused to participate or could not be contacted and two patients had died of unrelated causes. Thirty-six patients completed the trial with 18 from each treatment arm. MAIN OUTCOME MEASURES: Quality of voice as assessed by three questionnaires, Voice Handicap Index (VHI), Vocal Performance Questionnaire (VPQ), Voice Symptom Score (VoiSS) and perceptual analysis of voice by Grade, Roughness, Breathiness, Asthenia and Strained (GRBAS) assessment of vocal recordings. Quality of life as assessed by the Hospital Anxiety and Depression Scale (HADS), University of Washington Quality of Life Questionnaire (UW-QoL), and the Functional Assessment of Cancer Therapy (FACT) questionnaire. RESULTS: All patients included in the trial were able to complete the questionnaires; however, 19% required assistance of some kind. GRBAS assessment showed no difference between groups for any criteria. All QoL questionnaires gave equivalent good scores. All of the voice questionnaires showed no statistical difference between groups except for the emotional subscale of the VoiSS which showed a significantly better score for the radiotherapy arm (P = 0.04). CONCLUSION: All outcome measures were applicable and acceptable to the patient group. Overall QoL and voice appears similar despite treatment arm, apart from the emotional subscale of the VoiSS. A randomized controlled trial is required to further assess this question. PMID- 15748190 TI - Does thyroid subspecialization alter practice and outcome? A completed 4-year audit loop. AB - OBJECTIVE: To assess the effect of subspecialization on thyroid surgical practice and outcome. DESIGN: The multidisciplinary thyroid clinic was established in November 2000. A retrospective study of all thyroid surgery covering the period November 1998 to November 2000 was carried out. Best practice principles were implemented and prospective data were collected over a further 2 years to complete the audit loop. Practice and results were compared. SETTING: The project was carried out through the Hull and East Yorkshire Multidisciplinary Thyroid Clinic, a tertiary referral clinic for the management of thyroid disease. PARTICIPANTS: All patients who underwent thyroid surgery through the Department of Otolaryngology Head and Neck Surgery for 2 years before the establishment of the thyroid clinic (39 patients) and all patients who underwent thyroid surgery through the department for 2 years after the establishment of the clinic (108 patients) were included. MAIN OUTCOME MEASURES: Referral patterns, preoperative workup, types of surgery, histopathological diagnosis and complications rates were compared. RESULTS: The audit loop revealed that permanent vocal cord palsy rates had fallen from 8% to 0%, haematoma rates had fallen from 5% to 2% and permanent hypoparathyroidism rates had fallen from 8% to 2%. In addition, caseload had markedly increased, preoperative investigations were reduced and types of surgery standardized. CONCLUSION: It is clear that thyroid subspecialization is beneficial as far as overall outcome is concerned. However, this may lead to increased throughput which must be borne in mind for future activity planning. PMID- 15748192 TI - An important, randomized controlled trial of adenotonsillectomy. PMID- 15748191 TI - Treatment of hypopharyngeal carcinoma: analysis of nationwide study in the Netherlands over a 10-year period. AB - OBJECTIVE: To analyse different treatment strategies and treatment results of hypopharyngeal carcinoma in the Netherlands. DESIGN: Retrospective study. SETTING: Eight head and neck centres in the Netherlands. PARTICIPANTS: A total of 893 patients were treated between 1985 and 1994. Patients were mostly treated with radiotherapy alone, combined surgery and radiotherapy and surgery alone. RESULTS: The 5-year survival for the whole group was 26%. The 5-year survival for patients treated with curative intention was 32% and treated with palliative intention was 5%. The 5-year disease-free survival after radiotherapy alone was 37%, after surgery alone 41% and after combined therapy 47%. The role of chemotherapy could not be investigated because of a small number of patients treated with chemotherapy in this period. CONCLUSION: Combined therapy with surgery and radiotherapy has a better survival for patients with a hypopharyngeal carcinoma in comparison with radiotherapy alone. The N-stage is more important for the prognosis than the T-stage. PMID- 15748193 TI - Effectiveness of adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy: open, randomised controlled trial. AB - OBJECTIVE: To assess the effectiveness of adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy. DESIGN: Open, randomised controlled trial. SETTING: 21 general hospitals and three academic centres in the Netherlands. PARTICIPANTS: 300 children aged 2-8 years requiring adenotonsillectomy. INTERVENTION: Adenotonsillectomy compared with watchful waiting. MAIN OUTCOME MEASURES: Episodes of fever, throat infections, upper respiratory tract infections, and health related quality of life. RESULTS: During the median follow up period of 22 months, children in the adenotonsillectomy group had 2.97 episodes of fever per person year compared with 3.18 in the watchful waiting group (difference -0.21%, 95% confidence interval -0.54 to 0.12), 0.56 throat infections per year compared with 0.77 (-0.21, -0.36 to 0.06), and 5.47 upper respiratory tract infections per person year compared with 6.00 (-0.53, -0.97 to -0.08). No clinically relevant differences were found for health related quality of life. Adenotonsillectomy was more effective in children with a history of three to six throat infections than in those with none or two. 12 children had surgery related complications. CONCLUSION: Adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy has no major clinical benefits over watchful waiting. PMID- 15748194 TI - How we do it: radiofrequency-turbinectomy for nasal obstruction symptoms. AB - Radiofrequency energy has been reported to be effective to reduce nasal obstruction because of inferior turbinate hypertrophy. A total of 158 patients who had undergone such surgery are reported. Three to 30 months postoperatively 85% of the patients reported a marked decrease in nasal obstruction. This was irrespective of a short-term or a long-term follow-up, and was applicable to patients that had septal deviations in addition. Radiofrequency-turbinectomy is a simple and effective method for treatment of nasal obstruction as a result of hypertrophy of the inferior turbinates, even if combined with a moderate septal deviation. PMID- 15748195 TI - How we do it: voice therapy to improve vocal constitution and endurance in female student teachers. AB - Vocal hypofunction should be regarded as risk factor of occupational voice disorders. Voice therapy is effective to improve vocal endurance and voice constitution. Elucidation of constitutionally weak voices in teachers is recommended prior to their professional career. PMID- 15748196 TI - How we do it: blood contamination during management of epistaxis--awareness, utilization and availability of barrier protection. AB - There is a significant risk of blood contamination during the non-surgical management of epistaxis. Contamination beyond gloves happens in 55% of cases and the contamination risk to eyes as high as 18%. There is a lack of awareness and a majority of ENT doctors underestimate this risk. Protective barrier equipment and eye protection in particular are suboptimally available in all clinical areas where epistaxis is dealt with. Consequently, there is poor utilization of protective barrier equipment. PMID- 15748197 TI - How we think: debate on the aetiology of chronic sinusitis. PMID- 15748198 TI - Interobserver perceptual analysis of smokers voice. PMID- 15748199 TI - Use of disposable curette in the treatment of Chondrodermatitis Nodularis Helicis. PMID- 15748201 TI - Nasal fracture manipulation: a comparative study of general and local anaesthesia techniques. PMID- 15748202 TI - Endaural or postaural incision for myringoplasty: does it make a difference to the patient? PMID- 15748206 TI - Lipid based particulate formulations for the delivery of antigen. AB - Particulate adjuvant systems are largely classified according to their functional characteristics, such as the nature of the typical immune response they induce, or their perceived mode of action. From a formulation science perspective, it is practical to classify antigen delivery systems according to the physical nature of the formulations. This article discusses lipid based particulate systems, grouped according to the nature of their predominant lipid constituent. PMID- 15748207 TI - Histidine-rich glycoprotein: A novel adaptor protein in plasma that modulates the immune, vascular and coagulation systems. AB - Histidine-rich glycoprotein (HRG) is an abundant plasma glycoprotein that has a multidomain structure, interacts with many ligands, and has been shown to regulate a number of important biological processes. HRG ligands include Zn(2+) and haem, tropomyosin, heparin and heparan sulphate, plasminogen, plasmin, fibrinogen, thrombospondin, IgG, FcgammaR and complement. In many cases, the histidine-rich region of the molecule enhances ligand binding following interaction with Zn(2+) or exposure to low pH, conditions associated with sites of tissue injury or tumour growth. The multidomain nature of HRG indicates that it can act as an extracellular adaptor protein, bringing together disparate ligands, particularly on cell surfaces. HRG binds to most cells primarily via heparan sulphate proteoglycans, binding which is also potentiated by elevated free Zn(2+) levels and low pH. Recent reports have shown that HRG can modulate angiogenesis and additional studies have shown that it may regulate other physiological processes such as cell adhesion and migration, fibrinolysis and coagulation, complement activation, immune complex clearance and phagocytosis of apoptotic cells. This review outlines the molecular, structural, biological and clinical properties of HRG as well as describing the role of HRG in various physiological processes. PMID- 15748208 TI - ISCOM-based vaccines: the second decade. AB - The immunostimulating complex or 'iscom' was first described 20 years ago as an antigen delivery system with powerful immunostimulating activity. Iscoms are cage like structures, typically 40 nm in diameter, that are comprised of antigen, cholesterol, phospholipid and saponin. ISCOM-based vaccines have been shown to promote both antibody and cellular immune responses in a variety of experimental animal models. This review focuses on the evaluation of ISCOM-based vaccines in animals over the past 10 years, as well as examining the progress that has been achieved in the development of human vaccines based on ISCOM adjuvant technology. PMID- 15748210 TI - NK gene complex haplotype variability and host resistance alleles to murine cytomegalovirus in wild mouse populations. AB - The NK gene complex (NKC) on mouse chromosome 6 encodes receptors that are expressed on NK cells, such as Ly49H, and is involved in regulating NK cell control of virus infections, such as murine cytomegalovirus (MCMV). In the present study, we investigated the level of allelic heterogeneity in NKC loci in populations of outbred wild mice. This work revealed extensive levels of heterogeneity within two wild mouse populations. Analysis of MCMV replication in a population of specific pathogen-free outbred wild mice revealed that low viral titres, which are normally associated with the Cmv1(r) allele of the Cmv1 host resistance locus, were not prevalent in the mice tested. Hence, NKC-mediated resistance associated with Cmv1(r)/Ly49H-like effects was rare in this population. Overall, these data indicate that the NKC region is highly polymorphic and thus it is very likely that it confers on mice sufficient variability to cope with infection by a range of pathogens. PMID- 15748209 TI - Marked structural and functional heterogeneity in CXCR4: separation of HIV-1 and SDF-1alpha responses. AB - CXCR4, the chemotactic cell receptor for SDF-1alpha, is essential for immune trafficking and HIV infection. CXCR4 is remarkably heterogeneous and the purpose of this study was to better identify the isoforms expressed by cells and compare their structure and function. We found that cells express either a predominant isoform or multiple isoforms. These were best resolved on SDS-PAGE using sucrose gradient-fractionated, triton-insoluble, membrane extracts. We hypothesized that glycosyl modification may underpin some of this heterogeneity and that cell isoform(s) differences may underscore CXCR4's multiple cell functions. A comparison of wild-type (WT) and dual N-linked glycosylation site, N11A/N176A, mutant CXCR4 expressed in 3T3 and HEK-293 cells served to implicate variabilities in glycosylation and oligomerization in almost half of the isoforms. Immunoprecipitation of CXCR4 revealed monomer and dimer non-glycosylated forms of 34 kDa and 68 kDa from the N11A/N176A mutant, compared with glycosylated 40 kDa and 47 kDa and 73 kDa and 80 kDa forms from WT. The functional specificity of isoform action was also implicated because, despite CEMT4 cells expressing high levels of CXCR4 and 11 different isoforms, a single 83 kDa form was found to bind gp120 for HIV-1 IIIB infection. Furthermore, comparative studies found that in contrast to SDF-1alpha-responsive Nalm-6 cells that expressed similar levels of a single isoform, CEMT4 cells did not show a Ca(++) flux or a chemotactic response to SDF-1alpha. Thus, CXCR4 can differ both structurally and functionally between cells, with HIV-1 infection and chemotaxis apparently mediated by different isoforms. This separation of structure and function has implications for understanding HIV-1 entry and SDF-1alpha responses and may indicate therapeutic possibilities. PMID- 15748211 TI - Protein kinase A affects Galleria mellonella (Insecta: Lepidoptera) larval haemocyte non-self responses. AB - We used the protein kinase A (PKA) specific activator Sp-8-Br-cAMPS and type I inhibitor Rp-8-Br-cAMPS alone and in combination to define the role of PKA in the non-self responses of larval Galleria mellonella haemocytes in vitro and in vivo. Active PKA depressed haemocyte responses whereas PKA inhibition enhanced activities, including bacterial phagocytosis, the number of haemocytes with adherent bacteria, bacterial-induced haemocytic protein release and haemocyte adhesion to slides in vitro, as well as in vivo bacterial removal from the haemolymph. Non-attached haemocytes had more PKA activity than attached haemocytes; therefore, active PKA limited haemocyte response to foreign materials. We found that (i) PKA inhibitor alone induced non-self responses, including haemocyte protein discharge and lowered haemocyte counts in vivo, and induced nodulation; (ii) the enzyme activator produced effects opposite to those of the inhibitor; and (iii) together, the modulators offset each others' effects and influenced haemocyte lysate PKA activity. These findings establish PKA as a mediator of haemocytic non-self responses. PMID- 15748212 TI - Increased peripheral blood T-cell apoptosis and decreased Bcl-2 in chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is an inflammatory airway disease, usually associated with cigarette smoking. Stimulated peripheral blood T cells from patients with COPD have an increased propensity to undergo apoptosis. The mitochondrial apoptotic pathway is regulated by pro-apoptotic proteins (including p53 and Bax) as well as anti-apoptotic proteins (e.g. Bcl-2) and cytokines (IL-2, IL-4 and IL-7). We hypothesized that alterations in expression of these apoptosis related proteins, cytokines and cytokine receptors may be important in determining the susceptibility of T cells to undergoing apoptosis in COPD. We further hypothesized that inhaled corticosteroids (GCS) contribute to the increased rates of T-cell apoptosis observed in COPD. The process of apoptosis (assessed by Annexin V and ssDNA staining), as well as Bcl-2, Bax, p53, IL-2, IL 4 and receptors IL-7R, IL-4R and IL-2Rgamma were investigated in PHA-stimulated peripheral blood-derived T cells, using flow cytometry. Fifteen patients with COPD receiving inhaled GCS (four of who received additional prednisolone), eight patients with COPD receiving symptom control medication, and 16 control subjects were studied. T cells (CD4(+) and CD8(+)) from GCS-treated COPD patients showed an increased propensity to undergo apoptosis, associated with significantly decreased Bcl-2 and IL-7 receptor expression. No significant differences were observed for the COPD patients who were receiving symptom control medication. These findings may suggest a negative peripheral effect of inhaled GCS on the immune system in COPD, although the clinical significance of these effects remains uncertain. PMID- 15748213 TI - Fibronectin on activated T lymphocytes is bound to gangliosides and is present in detergent insoluble microdomains. AB - Fibronectin (FN) is a multifunctional extracellular matrix glycoprotein, which participates in cell migration and signalling to adhering cells. Due to alternative splicing and post-translational modifications, different isoforms of FN are generated by a wide variety of cells. T lymphocytes synthesize a surface associated isoform of FN, containing the two 'extradomains' EDA and EDB. In the present study, we identified gangliosides within the T-cell membrane as specific binding sites for the N-terminal 30 kDa fragment of FN. When T cells were activated with anti-CD3 coated beads, FN, together with the ganglioside GM1, converged at the contact zone. Moreover, endogenous FN was present in the detergent insoluble microdomain. The function of FN in T cells is still under investigation; however, its presence together with gangliosides at the activation site suggests participation in T-cell signalling. PMID- 15748214 TI - Cooperation between CD4 T helper cells is required for the generation of alloantigen-specific, IFN-gamma-producing human CD4 T cells. AB - We have used MLR to assess the cell interactions that enable human PBL stimulated with irradiated, allogeneic PBL to give rise to allospecific, IFN-gamma-producing CD4 T cells. We were able to generate alloantigen-specific, IFN-gamma-producing T cells from peripheral blood. The responder and stimulator cell numbers required for the optimal generation of such cells, enumerated using an enzyme-linked immunospot assay, were separately determined for various combinations of responders and stimulators. The number of antigen-specific, IFN-gamma-producing cells generated per 10(6) responder cells, seeded at the initiation of culture, is critically dependent on the density of the responder cells, with minimal generation of IFN-gamma-producing T cells at low responder densities. These and further observations with fractionated responding PBL show that CD4 T-cell/CD4 T cell interactions, which are completely independent of CD8() T cells, are necessary for the in vitro generation of alloantigen-specific, IFN-gamma producing CD4 T cells. PMID- 15748215 TI - Does memory improve with age? CD85j (ILT-2/LIR-1) expression on CD8 T cells correlates with 'memory inflation' in human cytomegalovirus infection. AB - CMV-specific memory CD8(+) T cells accumulate over time to reach high frequencies amongst peripheral blood lymphocytes - a phenomenon termed 'memory inflation'. Using tetramer staining on samples from a large number of subjects and multivariate regression analysis, we were able to relate this to the phenotype of CD8(+) T cells. We made the following observations: (i) CD85j (ILT-2/LIR-1) was highly expressed alongside CD57 - an established effector memory marker - on CMV specific CD8(+) T cells; (ii) on CD8(+) T cells as a whole, with increasing age, CD57 and CD85j (ILT-2/LIR-1) expression increased whereas CCR7 expression decreased, indicating increasing maturation of the total CD8(+) T-cell compartment with age; (iii) unit increases in the percentage of CMV-specific CD8(+) T cells expressing CD57 and CD85j (ILT-2/LIR-1) were associated with incremental expansion of these T-cell populations; (iv) CMV seropositivity is associated with a marked effect on the overall phenotype of CD8(+) T cells (at any given age, CMV seropositivity is associated with an 18.7% increase in CD85j (ILT-2/LIR-1) expression); and (v) from our observations we estimated from this an apparent 'ageing effect' of CMV on CD8(+) T cells of 35.4 years. The data presented are consistent with a predictable, unidirectional and linear model of virus specific T-cell differentiation and maturation. PMID- 15748216 TI - Blockade of IFN-gamma does not affect the arthritogenicity of T cells generated during the induction of adjuvant arthritis but exacerbates the polyarthritis produced by adoptive transfer of arthritogenic effector cells. AB - IFN-gamma production is prominent in some models of autoimmune disease, including adjuvant arthritis (AA), but the role of IFN-gamma in the pathogenesis of these diseases is uncertain. Experimental manipulation (administration of cytokine, blocking cytokine action with specific antibody, disruption of genes encoding the cytokine or its receptor) has revealed both pro- and anti-inflammatory effects, depending on the nature of the manipulation and the timing of the treatment. We examined separately the effects of cytokine blockade during the afferent and efferent phases of AA in Dark Agouti rats, using an adoptive transfer system. Effects of IFN-gamma on the efferent phase were investigated by treating recipients with mAb DB-1, which blocks the activity of rat IFN-gamma. When treatment was commenced before cell transfer, the resulting polyarthritis was more severe than in controls treated with normal IgG. Commencing treatment after the adoptively transferred disease had become established caused neither amelioration nor exacerbation, but did cause some delay in resolution. In contrast, treatment of donors did not appear to affect the generation of arthritogenic cells. The main effect of IFN-gamma appears to be modulation of the arthritogenicity of the migratory effector T cells that can transfer AA. PMID- 15748217 TI - Evidence of Toll-like receptor molecules on human platelets. AB - Platelets are primarily involved in thrombosis and haemostasis, and they have recently been shown to have a role in innate immunity and in inflammation. We have determined the markers of innate immunity that are expressed by platelets, specifically the Toll-like receptors (TLR), originating from mixes of platelet concentrates (MPC, n = 5) between day zero and day five after blood collection. The surface membrane and intracellular expression of TLR were measured, both after and without permeabilization, using flow cytometry. We observed weak expression of TLR2, TLR4 and TLR9 on the surface of CD41(+) platelets. The expression levels of TLR4 were high (59 +/- 2.2%). Moreover, there was a significant expression of TLR2 (47.5 +/- 4.8%), TLR4 (78.8 +/- 1.3%) and TLR9 (34.2 +/- 7.5%) in the cytoplasm of CD41(+) platelets. The expression of the three receptors did not change significantly during the course of the 5 day observation period. The percentage of TLR expression is significantly modulated between activated versus non-activated platelets, both after and without permeabilization (P < 0.01). Study of the expression of TLR could increase our knowledge of the level of platelet participation during an immune reaction and inflammation. In the same way as the platelet ligand/receptor pair CD40L/CD40 is, the TLR are expressed by platelets, and could serve as a link between innate and adaptive immunity. PMID- 15748220 TI - Evaluation of interactive online courses for advanced practice nurses. AB - PURPOSE: The purpose of this study was twofold: (a) to examine the effectiveness of two interactive online learning modules for advanced practice nurses (APNs) and (b) to examine the participants' demographic characteristics and their perceptions of the strengths and weaknesses of online learning. DATA SOURCES: A purposive sample of 73 registered nurses and APNs enrolled in graduate study or continuing education completed an online learning course. These participants were pretested and post-tested using a knowledge test developed by the authors. Participants were also surveyed for demographic characteristics and perceptions toward online learning. CONCLUSIONS: The findings of this study support case based online learning as a successful method in the education of APNs. The evaluation of the online modules and the enthusiasm from students indicated success of this teaching/learning method. The majority of learners responded positively toward the online method of learning and included comments that indicated they enjoyed learning using this method, would like to have the site available at their practice locations for reference, and would like to have other courses designed and delivered in this manner. IMPLICATIONS FOR PRACTICE AND EDUCATION: The effectiveness and quality of the online instruction ultimately matter most to student satisfaction in online courses. The future of online education is dependent on well-structured, interactive, and substantive programs. Educators are challenged to continually assess and evaluate the changing learning needs of APNs. PMID- 15748221 TI - Homocysteine and cardiovascular disease: a review of current recommendations for screening and treatment. AB - PURPOSE: To review the literature for, and provide advanced practice nurses (APNs) with, current recommendations for screening and treatment of hyperhomocysteinemia. DATA SOURCES: Medscape literature search of selected research studies and related journal articles. CONCLUSIONS: While data from most epidemiologic studies support the argument that hyperhomocysteinemia is an independent risk factor for cardiovascular disease, the debate continues as to when screening and treating patients is appropriate. The consensus is that more randomized controlled trials are needed to further study the benefits of routine screening and the efficacy of treating hyperhomocysteinemia. IMPLICATIONS FOR PRACTICE: Until the results of ongoing clinical trials are available, APNs should follow the American Heart Association guidelines for screening for elevated levels of homocysteine and continue to promote a well-balanced diet that includes foods rich in folic acid as part of health promotion through primary prevention. PMID- 15748222 TI - Recognizing health risks in childhood cancer survivors. AB - PURPOSE: To provide information about the special health needs of childhood cancer survivors through a discussion of Wilms' tumor, a common childhood renal tumor, and case studies of two survivors of Wilms' tumor, who encountered significant health problems as adults. This information may be generalized to many cancer survivors. DATA SOURCES: Actual case studies as well as extensive review of the current literature on childhood cancer survivorship. CONCLUSIONS: Adult healthcare practitioners may help to decrease morbidity of childhood cancer survivors in adulthood when they are knowledgeable about the risks of this population. Resources are readily available, both in the medical literature and on the Internet. IMPLICATIONS FOR PRACTICE: With well-established success in the treatment of childhood cancer, there are many young and middle-aged adult survivors of the disease. However, as many as two-thirds of survivors may have chronic long-term effects of treatment. Most survivors eventually discontinue follow-up with their pediatric oncologists and receive primary health care from adult providers, many of whom are unaware of survivors' health risks. Awareness of primary care providers regarding the special health needs of childhood cancer survivors is critical to decrease morbidity and improve quality of life. The ever increasing number of childhood cancer survivors is creating a public health issue as they transition into adulthood. PMID- 15748223 TI - When cure is care: diagnosis and management of pulmonary arterial hypertension. AB - PURPOSE: The purpose of this article is to provide nurse practitioners with an understanding of the pathophysiology of pulmonary arterial hypertension (PAH) disease, clinical manifestations, diagnostic evaluation, drug therapy, strategies for health promotion, and relevant care issues for patients and families. DATA SOURCES: Selected clinical and research articles, as well as current government guidelines. CONCLUSIONS: Symptoms expressed are more apparent as PAH disease progresses, leaving fewer treatment options in advanced disease stages. New drugs are currently being tested for the treatment of PAH; however, the costs of many of the currently approved treatments may be prohibitive. IMPLICATIONS FOR PRACTICE: Earlier recognition of disease symptoms leads to prompt initiation of diagnostic evaluation and referral to specializing medical centers. Upon referral, specialty centers may begin appropriate treatment regimens earlier in the disease process, which could improve clinical outcomes and quality of life. PMID- 15748224 TI - Osteoporosis risk and interest in strength training in men receiving androgen ablation therapy for locally advanced prostate cancer. AB - PURPOSE: The overall purpose of this preliminary study to a clinical trial is to explore the feasibility of recruiting men receiving androgen ablation therapy (AAT) for locally advanced prostate cancer to a future strength training study for the prevention of osteoporosis. The threefold specific purpose of this comparative and correlational study is to (a) describe the prevalence of risk factors for osteoporosis, (b) compare functional status and symptom distress between those interested and not interested in a future strength training study, and (c) examine relationships among self-efficacy for strength training, functional status, symptom distress, years since cancer diagnosis, and cumulative dose of AAT in this at-risk population. METHOD: Data were obtained from 40 Caucasian men (mean age = 75.8 years) with locally advanced prostate cancer, capable of self-care, and receiving AAT. The European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaires (QLQ) EORTC QLQ C30 and EORTC QLQ-PR25 (prostate cancer specific) were used to measure functional status and symptom distress. A 6-item, investigator-developed questionnaire was used to measure self-efficacy for learning and overcoming barriers to strength training. CONCLUSIONS: The majority of these men had multiple risk factors for osteoporosis, in addition to receiving AAT, and had not completed bone mineral density (BMD) testing. A high percentage (67.5%) expressed interest in a future strength training protocol for osteoporosis prevention; functional status and symptom distress did not appear to influence this interest. Several aspects of functional status and symptom distress were significantly related to self efficacy for learning and overcoming barriers to strength training. Years since cancer diagnosis and cumulative dose of AAT were not significantly related to self-efficacy for strength training, but age was related. This study informs a planned future experimental study by establishing the recruitment potential and the decreased likelihood of symptom distress interfering with self-efficacy for participation in strength training. IMPLICATIONS FOR PRACTICE: Advanced practice nurses should complete assessments for osteoporosis risk factors, monitor height loss, order BMD testing and bone enhancing medications, and discuss lifestyle modifications including strength training to improve bone health in men receiving AAT for prostate cancer. PMID- 15748225 TI - Uncertain times for research on hemophilia and allied disorders. PMID- 15748226 TI - Coley's toxin revisited: immunotherapy or plasminogen activator therapy of cancer? PMID- 15748227 TI - Antithrombotic strategies for atrial fibrillation: on the threshold of changes? Yes. PMID- 15748228 TI - Antithrombotic strategies for atrial fibrillation: on the threshold of changes? No. PMID- 15748229 TI - Factor Xa inhibitors in acute coronary syndromes: moving from mythology to reality. PMID- 15748230 TI - First experience with direct, selective factor Xa inhibition in patients with non ST-elevation acute coronary syndromes: results of the XaNADU-ACS Trial. AB - BACKGROUND: Unfractionated heparin is widely used in patients with non-ST elevation acute coronary syndromes but has important limitations. Anticoagulants with predictable kinetics and anticoagulant effects, better efficacy, and greater safety are needed. OBJECTIVE: To investigate the efficacy and safety of a direct, selective factor Xa inhibitor, DX-9065a (Daiichi Pharmaceuticals LTD, Inc.) compared with heparin, in patients with non-ST-elevation acute coronary syndromes. PATIENTS AND METHODS: Patients (n = 402) from the USA, Canada, and Japan were randomized to blinded, weight-adjusted heparin, low-dose DX-9065a, or high-dose DX-9065a. RESULTS: The primary efficacy endpoint of death, myocardial infarction, urgent revascularization, or ischemia on continuous ST-segment monitoring occurred in 33.6%, 34.3%, and 31.3% of patients assigned to heparin, low-dose DX-9065a, and high-dose DX-9065a (P = 0.91 for heparin vs. combined DX 9065a). The composite of death, myocardial infarction, or urgent revascularization occurred in 19.5%, 19.3%, and 11.9% (P = 0.125 for heparin vs. high-dose DX-9065a) of patients; major or minor bleeding occurred in 7.7%, 4.2%, and 7.0% of patients; and major bleeding in 3.3%, 0.8%, and 0.9% of patients. Higher concentrations of DX-9065a were associated with a lower likelihood of ischemic events (P = 0.03) and a non-significant tendency toward a higher likelihood of major bleeding (P = 0.32). CONCLUSIONS: In this small phase II trial, there was a non-significant tendency toward a reduction in ischemic events and bleeding with DX-9065a compared with heparin in patients with acute coronary syndromes. The absence of an effect on ST-monitor ischemia warrants further investigation. These data provide the rationale for adequately powered studies of DX-9065a in acute coronary syndromes or percutaneous intervention. PMID- 15748231 TI - One more way that mice and men are different. PMID- 15748232 TI - Megakaryocytes endocytose and subsequently modify human factor V in vivo to form the entire pool of a unique platelet-derived cofactor. AB - Factor Va (FVa), derived from plasma or released from stimulated platelets, is the essential cofactor in thrombin production catalyzed by the prothrombinase complex. Plasma-derived factor V (FV) is synthesized in the liver. The source(s) of the platelet-derived cofactor remains in question. We identified a patient homozygous for the FV(Leiden) mutation, who received a liver transplant from a homozygous wild-type FV donor. Eighteen days post-transplant, phenotypic analysis of the patient's platelet-derived FV indicated that the platelets were acquiring wild-type FV, consistent with the temporal differentiation of megakaryocytes and subsequent platelet production. Nine months post-transplant, the platelet-derived FV pool consisted entirely of wild-type FV. Consequently, megakaryocyte endocytosis of plasma-derived FV must account for the entire platelet-derived pool, because blood-borne platelets cannot bind or endocytose FV. Subsequent to this endocytic process, the patient's platelet-derived FV was cleaved to a partially active cofactor, and rendered resistant to phosphorylation catalyzed by a platelet-associated kinase, and hence less susceptible to activated protein C catalyzed inactivation. These data provide the first in vivo demonstration of an endocytosed plasma protein undergoing intracellular modifications that alter its function. This process results in the sequestration of active FVa within the platelet compartment, poised for immediate action subsequent to release from platelets at a site of injury. PMID- 15748233 TI - Asymptomatic thrombophilia--a family affair. PMID- 15748235 TI - Vessel wall apoptosis and atherosclerotic plaque instability. AB - Atherosclerotic cardiovascular disease remains the leading cause of death in the industrialized world. Most cardiovascular deaths result from acute coronary syndromes, including unstable angina pectoris and acute myocardial infarction. Coronary syndromes often arise from acute coronary thrombosis, itself commonly a result of disruption or rupture of the fibrous cap of a lipid-laden atherosclerotic plaque. Despite this huge clinical burden of atherosclerotic plaque instability, our understanding of the molecular mechanisms mediating atherosclerotic plaque disruption and rupture, at a cellular level, remains limited. Placed in a clinical context, this review discusses our current understanding of the molecular basis for atherosclerotic plaque instability, with particular emphasis on the process of apoptosis-or programmed cell death-seen increasingly as playing a key role in a number of cell types within the vessel wall. PMID- 15748234 TI - Risk of a first venous thrombotic event in carriers of a familial thrombophilic defect. The European Prospective Cohort on Thrombophilia (EPCOT). AB - BACKGROUND: Reliable risk estimates for venous thrombosis in families with inherited thrombophilia are scarce but necessary for determining optimal screening and treatment policies. OBJECTIVES: In the present analysis, we determined the risk of a first venous thrombotic event in carriers of a thrombophilic defect (i.e. antithrombin-, protein C- or protein S deficiency, or factor V Leiden). PATIENTS AND METHODS: The asymptomatic carriers had been tested prior to this study in nine European thrombosis centers because of a symptomatic carrier in the family, and were followed prospectively for 5.7 years on average between March 1994 and January 2001. Annually, data were recorded on the occurrence of risk situations for venous thrombosis and events (e.g. venous thrombosis, death). RESULTS: Twenty-six of the 575 asymptomatic carriers (4.5%) and seven of the 1118 controls (0.6%) experienced a first deep venous thrombosis or pulmonary embolism during follow-up. Of these events, 58% occurred spontaneously in the carriers compared with 43% in the controls. The incidence of first events was 0.8% per year (95% CI 0.5-1.2) in the carriers compared with 0.1% per year (95% CI 0.0-0.2) in the controls. The highest incidence was associated with antithrombin deficiency or combined defects, and the lowest incidence with factor V Leiden. CONCLUSIONS: The incidence of venous events in asymptomatic individuals from thrombophilic families does not exceed the risk of bleeding associated with long-term anticoagulant treatment in the literature (1 3%). PMID- 15748236 TI - Primary and secondary prophylaxis of venous thromboembolism with low-molecular weight heparins: prolonged thromboprophylaxis, an alternative to vitamin K antagonists. AB - Low-molecular-weight heparins (LMWHs) are used widely in the treatment and prevention of venous thromboembolism (VTE). The LMWHs dalteparin and enoxaparin reduce the rate of VTE by at least 50% if administered for 4-5 weeks following major orthopedic surgery, compared with in-hospital prophylaxis for 7-15 days. Meta-analyses have confirmed that the size of the reduction is similar for both clinical and asymptomatic VTE. Vitamin K antagonists (VKAs) have been shown to be associated with significantly higher bleeding rates compared with LMWH when used as prolonged prophylaxis against VTE following major orthopedic surgery. Patients with cancer are a recognized group at high risk of VTE, and those undergoing major surgery for their malignancy are at particular risk. Evidence from clinical trials is amassing to show that prolonged prophylaxis with LMWH (dalteparin, enoxaparin) in these patients can significantly reduce the rate of postoperative VTE. In cancer patients with acute VTE, the traditional approach is to initiate acute treatment with unfractionated heparin or LMWH followed by long-term treatment with VKA to prevent recurrence. However, clinical trial data have confirmed that the LMWH dalteparin, when administered for 6 months, is significantly more effective than VKA in preventing recurrence, cutting the rate of VTE by 52% without increasing the risk of bleeding. A new and intriguing area of interest is whether LMWH can enhance survival in patients with cancer. Preliminary data suggest that a biological effect of LMWH may act to prolong survival in patients with cancer. PMID- 15748237 TI - Mutations in GPIIIa molecule as a cause for Glanzmann thrombasthenia in Indian patients. AB - BACKGROUND: Glanzmann thrombasthenia (GT) results from a quantitative or qualitative defect of GPIIb-IIIa complex, the fibrinogen receptor on platelets, which plays a very important role in platelet aggregation. In this report we describe the molecular studies on 22 patients with Glanzmann Thrombasthenia at our institute. OBJECTIVES: The main objective was to identify the mutations present in our GT population in order to establish a strategy for genetic counseling and antenatal diagnosis. METHODS: Twenty-two patients with GT were included in the present study. Complete blood count (CBC), platelet aggregation, flow cytometry, Western blot, single strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) were performed in all the patients. The patients showing an abnormal migration pattern in SSCP or DGGE were sequenced further on an automated sequencer. RESULTS: Of the 22 patients studied, mutations were detected in 12 individuals. Of these, 11 were novel mutations and one mutation Y115C was reported earlier. Flow cytometric analysis showed the absence of receptors in type I GT, highly reduced levels in type II GT and normal levels in type III GT. The DGGE analysis and SSCP analysis of the patients showed different migration patterns. Sequencing was performed in all patients showing an abnormal migration pattern. Of the 22 cases studied mutations could be detected in 12 cases of GT. We could detect six patients with point mutations, four patients with insertions and five patients with deletion mutations. Exon 4 has been found to be the most common site for mutations in our patients. CONCLUSION: This study has shown a wide array of mutations present in our GT patients which would be extremely useful in genetic counseling and prenatal diagnosis, essential in preventing these disorders in succeeding generations. PMID- 15748238 TI - Insertion of a C in the exon 28 of integrin alphaIIb gene leading to a frameshift mutation is responsible for Glanzmann thrombasthenia in a Japanese case. AB - BACKGROUND: Glanzmann thrombasthenia (GT) is a hereditary bleeding disorder caused by a qualitative or quantitative defect in the integrin alphaIIbbeta3. OBJECTIVE: Our objective is to identify the gene mutation that resulted in GT. PATIENTS AND METHODS: The patient was a 66-year-old male with a history of frequent bleeding. The expression levels of the integrin proteins in the platelets were determined by flow cytometry and Western blot analysis. The sequences of genomic DNA and mRNA encoding for alphaIIb and beta3 were analyzed by the dye-terminator cycle sequencing method. For transfection experiments, expression vectors encoding for wild-type alphaIIb, mutated alphaIIb, beta3, green fluorescent protein (GFP) fusion wild-type alphaIIb, GFP fusion mutated alphaIIb and DsRed fusion beta3 were constructed. These vectors were transfected to COS-7 cells, and the expression levels were determined. RESULTS: The alphaIIb protein was remarkably reduced in the patient's platelets, and gene analysis showed that the patient possessed compound heterozygous mutations in the alphaIIb gene. One was a C --> G substitution at the splice acceptor site (- 3) of exon 26 (CAG -->GAG) and the other was the insertion of an additional C at the region including six C bases between 2911 and 2916 in exon 28 (InsC). Transfection experiments using COS-7 cells showed that alphaIIb containing InsC had expressed and formed a complex with beta3, but had not been transported to the Golgi apparatus. CONCLUSIONS: In the present study the novel mutation InsC, leading to a frameshift that affects the transmembrane domain and the cytoplasmic tail, was found to be responsible for GT. PMID- 15748239 TI - Protein Z, protein S levels are lower in patients with thrombophilia and subsequent pregnancy complications. AB - OBJECTIVE: We posit that low levels of protein S (PS) and protein Z (PZ) contribute to adverse pregnancy outcome (APO). PATIENTS: We evaluated 103 women with subsequent normal pregnancy outcome (NPO), 106 women with APO, and 20 women with thrombophilia (TP). METHODS: We compared first trimester (1st TRI) PZ levels in 103 women with NPO, 106 women with APO, and in 20 women with TP. We compared plasma levels of PZ and free PS antigen during the second (2nd TRI) and third trimesters (3rd TRI) of pregnancy in 51 women with APO and 51 matched women with NPO. RESULTS: The mean 1st TRI PZ level was significantly lower among patients with APO, compared to pregnant controls (1.81 +/- 0.7 vs. 2.21 +/- 0.8 microg mL( 1), respectively, P < 0.001). Of patients with known TP, those with APO had a tendency for lower mean PZ levels compared to those TP women with NPO (1.5 +/- 0.6 vs. 2.3 +/- 0.9 microg mL(-1), respectively, P < 0.0631). There was a significant decrease in the PZ levels in patients with APO compared to NPO (2nd TRI 1.5 +/- 0.4 vs. 2.0 +/- 0.5 microg mL(-1), P < 0.0001; and 3rd TRI 1.6 +/- 0.5 vs. 1.9 +/- 0.5 microg mL(-1), P < 0.0002). Protein S levels were significantly lower in the 2nd and 3rd TRIs among patients with APO compared to patients with NPO (2nd TRI 34.4 +/- 11.8% vs. 38.9 +/- 10.3%, P < 0.05, respectively; and 3rd TRI 27.5 +/- 8.4 vs. 31.2 +/- 7.4, P < 0.025, respectively). CONCLUSIONS: We posit that decreased PZ and PS levels are additional risk factors for APO. PMID- 15748240 TI - Thrombophilic risk factors in patients with severe carotid atherosclerosis. AB - Carotid stenosis and atrial fibrillation are the strongest risk factors for ischemic stroke. Ongoing prevention efforts include the identification of novel factors that increase the risk for carotid atherosclerosis. The aim of this study was to determine the thrombophilic risk profile of patients with severe carotid stenosis by evaluating a number of genetic and metabolic risk factors [factor (F)II G20210A, factor V Leiden, MTHFR C677T polymorphisms, anticardiolipin antibodies (aCL), lipoprotein(a) (Lp(a)), and homocysteine (Hcy)]. The study population consisted of 615 patients [(410 M/205 F; median age 73 (26-94) years] with severe (> 70%) carotid stenosis, and 615 apparently healthy subjects [(410 M/205 F; age 73 (31-92) years]. On multivariate analysis, independent risk factors were elevated Hcy [odds ratio (OR) 7.6, 95% confidence interval (CI) 4.8, 11.8] and Lp(a) levels (OR 2.9, 95% CI 2.1, 3.9), the presence of aCL (OR 5.7, 95% CI 3.1, 10.4) and heterozygosity for FII G20210A polymorphism (OR 2.8, 95% CI 1.3, 5.9). In the subgroup of women, independent risk factors for severe carotid atherosclerosis were: high levels of Hcy and Lp(a) and the presence of aCL, whereas hyperhomocysteinemia, elevated Lp(a) levels, aCL, FII G20210A and MTHFR 677TT polymorphisms remained independent risk factors in the subgroup of men. The results of the present study demonstrate that the prevalence of the thrombophilic risk factors is increased in patients with severe carotid atherosclerosis. PMID- 15748241 TI - The incidence and prognostic significance of elevated cardiac troponins in patients with submassive pulmonary embolism. AB - Although the incidence and prognostic significance of elevated cardiac troponins are known in patients with massive pulmonary embolism (PE), few studies have addressed this issue in patients with hemodynamically stable, submassive PE, who comprise the majority of patients presenting with PE. This prospective cohort study was, therefore, designed to determine the incidence and prognostic significance of elevated cardiac troponins in patients with submassive PE. Consecutive patients with acute, symptomatic, submassive PE that was confirmed by objective diagnostic testing were studied. All patients received treatment with either unfractionated heparin or fondaparinux followed by a coumarin derivative and underwent clinical follow-up for 3 months. Cardiac troponin I (cTnI) levels were measured within 24 h of clinical presentation. An elevated cTnI was defined as > 0.5 microg L(-1) and indicated myocardial injury. Major myocardial injury, that is associated with myocardial infarction, was defined by a cTnI > 2.3 microg L(-1). The clinical outcomes were recurrent venous thromboembolism and all-cause death. In 458 patients with submassive PE, the incidence of cTnI > 0.5 microg L( 1) was 13.5%[95% confidence interval (CI): 10.4-16.7], and the incidence of cTnI > 2.3 microg L(-1) was 3.5% (95% CI: 2.0-5.6). An elevated cTnI > 0.5 microg L( 1) was associated with an increased risk of all-cause death [odds ratio (OR) = 3.5; 95% CI: 1.0-11.9], but did not appear to confer an increased risk of recurrent venous thromboembolism (OR = 1.1; 95% CI: 0.2-4.9). In patients who present with submassive PE, an elevated cTnI occurs in about one in seven patients and is associated with a 3.5-fold increased risk of all-cause death. PMID- 15748242 TI - In vitro and in vivo studies of the novel antithrombotic agent BAY 59-7939--an oral, direct Factor Xa inhibitor. AB - BAY 59-7939 is an oral, direct Factor Xa (FXa) inhibitor in development for the prevention and treatment of arterial and venous thrombosis. BAY 59-7939 competitively inhibits human FXa (K(i) 0.4 nm) with > 10 000-fold greater selectivity than for other serine proteases; it also inhibited prothrombinase activity (IC(50) 2.1 nm). BAY 59-7939 inhibited endogenous FXa more potently in human and rabbit plasma (IC(50) 21 nm) than rat plasma (IC(50) 290 nm). It demonstrated anticoagulant effects in human plasma, doubling prothrombin time (PT) and activated partial thromboplastin time at 0.23 and 0.69 microm, respectively. In vivo, BAY 59-7939 reduced venous thrombosis (fibrin-rich, platelet-poor thrombi) dose dependently (ED(50) 0.1 mg kg(-1) i.v.) in a rat venous stasis model. BAY 59-7939 reduced arterial (fibrin- and platelet-rich) thrombus formation in an arteriovenous (AV) shunt in rats (ED(50) 5.0 mg kg(-1) p.o.) and rabbits (ED(50) 0.6 mg kg(-1) p.o.). Slight inhibition of FXa (32% at ED(50)) reduced thrombus formation in the venous model; to affect arterial thrombosis in the rat and rabbit, stronger inhibition of FXa (74%, 92% at ED(50)) was required. Calculated plasma levels in rabbits at the ED(50) were 14-fold lower than in the rat AV shunt model, correlating with the 14-fold lower IC(50) of FXa inhibition in rabbit compared with rat plasma; this may suggest a correlation between FXa inhibition and antithrombotic activity. Bleeding times in rats and rabbits were not significantly affected at antithrombotic doses (3 mg kg(-1) p.o., AV shunt). Based on these results, BAY 59-7939 was selected for clinical development. PMID- 15748243 TI - Does activated protein C-resistant factor V contribute to thrombin generation in hemophilic plasma? AB - In this study we assessed the role of factor V (FV) inactivation in hemophilic plasma with particular reference to the activated protein C (APC)-resistant variants FV-R506Q (FV Leiden) and FV-R306T (FV Cambridge). Purified recombinant full-length FV carrying these single substitutions and FV-R306T/R506Q were used in thrombin generation experiments. Plasma was first immunodepleted of FV, and subsequently of factors VIII, IX, or combinations thereof. Thrombin generation was initiated by low concentrations of recombinant tissue factor. Recombinant soluble thrombomodulin (TM) was used to trigger the APC system. Surprisingly, TM concentrations that reduced thrombin generation in normal plasma by no more than 50% virtually abolished thrombin formation in plasma deficient in the factor VIII/IX complex. This was already apparent at TM levels as low as 0.1 nmol L(-1). By varying the concentrations of purified (activated) protein C to plasma that was additionally depleted of protein C, we confirmed that impaired thrombin generation indeed was the result of the action of APC. In contrast, this did not occur when FV-depleted plasma had been reconstituted with FV-R306T/R506Q. Addition of FV-R306T or FV-R506Q partially reduced prothrombin activation, demonstrating the involvement of both APC cleavage sites. FV inactivation also occurred on the surface of human microvascular endothelial cells. Apparently, these cells express sufficient TM to down-regulate thrombin production via the APC pathway. We further conclude that in hemophilic plasma this pathway can induce a secondary defect because of premature FV inactivation. It therefore seems conceivable that APC-resistant FV has the potential of alleviating hemophilic bleeding. PMID- 15748244 TI - The synthetic pentasaccharide fondaparinux reduces coagulation, inflammation and neutrophil accumulation in kidney ischemia-reperfusion injury. AB - Ischemia-reperfusion (I/R) injury is associated with activation of coagulation and inflammation. Interestingly, various anticoagulants have been shown to reduce both coagulation and inflammation in animal models of kidney I/R injury. Fondaparinux is a synthetic pentasaccharide that selectively inhibits factor Xa (FXa) in the coagulation cascade. The aim of this study was to investigate the effect of fondaparinux in a lethal murine model of kidney I/R injury. A murine model of kidney I/R was established. In this model, we measured activation of the coagulation cascade and induction of inflammation. Administration of fondaparinux to I/R-injured mice reduced fibrin deposition in the kidney, reduced serum creatinine levels and increased survival from 0 to 44% compared with saline treated control mice. Fondaparinux also reduced interleukin-6 and macrophage inflammatory protein-2 expression and decreased neutrophil accumulation in the injured kidneys. Finally, we showed that fondaparinux reduced thioglycollate induced recruitment of neutrophils into the peritoneum and inhibited the binding of U937 cells to P-selectin in vitro. Our data suggest that fondaparinux reduces kidney I/R injury primarily by inhibiting the recruitment of neutrophils. PMID- 15748245 TI - Endocytosis of plasma-derived factor V by megakaryocytes occurs via a clathrin dependent, specific membrane binding event. AB - Megakaryocytes were analyzed for their ability to endocytose factor V to define the cellular mechanisms regulating this process. In contrast to fibrinogen, factor V was endocytosed by megakaryocytes derived from CD34(+) cells or megakaryocyte-like cell lines, but not by platelets. CD41(+)ex vivo-derived megakaryocytes endocytosed factor V, as did subpopulations of the megakaryocyte like cells MEG-01, and CMK. Similar observations were made for fibrinogen. Phorbol diester-induced megakaryocytic differentiation of the cell lines resulted in a substantial increase in endocytosis of both proteins as compared to untreated cells that did not merely reflect their disparate plasma concentrations. Factor IX, which does not associate with platelets or megakaryocytes, was not endocytosed by any of the cells examined. Endocytosis of factor V by megakaryocytes proceeds through a specific and independent mechanism as CHRF-288 cells endocytosed fibrinogen but not factor V, and the presence of other plasma proteins had no effect on the endocytosis of factor V by MEG-01 cells. Furthermore, as the endocytosis of factor V was also demonstrated to occur through a clathrin-dependent mechanism, these combined data demonstrate that endocytosis of factor V by megakaryocytes occurs via a specific, independent, and most probably receptor-mediated, event. PMID- 15748246 TI - The von Willebrand factor self-association is modulated by a multiple domain interaction. AB - BACKGROUND: Platelet adhesion and aggregation at sites of vascular injury exposed to rapid blood flow require von Willebrand factor (VWF). VWF becomes immobilized by binding to subendothelial components or by a self-association at the interface of soluble and surface-bound VWF. OBJECTIVES: As this self-association has been demonstrated only under shear conditions, our first goal was to determine whether the same interaction could be observed under static conditions. Furthermore, we wanted to identify VWF domain(s) important for this self-association. RESULTS: Biotinylated VWF (b-VWF) interacted dose-dependently and specifically with immobilized VWF in an enzyme-linked immunosorbent assay (ELISA) assay, showing that shear is not necessary to induce the VWF self-association. Whereas anti-VWF monoclonal antibodies (mAbs) had no effect on the self-association, the proteolytic VWF-fragments SpII(1366-2050) and SpIII(1-1365) inhibited the b-VWF VWF interaction by 70 and 80%, respectively. Moreover, a specific binding of b VWF to immobilized Sp-fragments was demonstrated. Finally, both biotinylated SpII and SpIII were able to bind specifically to both immobilized SpII and SpIII. Similar results were observed under flow conditions, which confirmed the functional relevance of our ELISA system. CONCLUSION: We have developed an ELISA binding assay in which a specific VWF self-association under static conditions can be demonstrated. Our results suggest a multiple domain interaction between immobilized and soluble VWF. PMID- 15748248 TI - Laboratory monitoring of low-molecular-weight heparin therapy--part II. Monitoring LMWH therapy? For the moment a non-question. PMID- 15748247 TI - Platelets adhered to endothelial cell-bound ultra-large von Willebrand factor strings support leukocyte tethering and rolling under high shear stress. AB - Leukocyte rolling on vascular endothelium is mediated by an interaction between P selectin expressed on endothelial cells and P-selectin glycoprotein ligand-1 on leukocytes. This interaction reduces the velocity of leukocyte movements to allow subsequent firm adhesion and transmigration. However, the interaction has so far been observed only under low venous shear stress and cannot explain the accumulation of monocytes in atherosclerotic plaques found in arteries, where shear stress is much higher. We have previously shown that newly released ultra large von Willebrand factor (ULVWF) forms extremely long string-like structures to which platelets tether. Here, we investigated whether platelets adhered to ULVWF strings are activated and form aggregates. We also determined whether activated platelets on ULVWF strings can support leukocyte tethering and rolling under high shear stresses. We found that platelets adhered to ULVWF expressed P selectin and bound PAC-1, suggesting their rapid activation. We also found that leukocytes tethered to and rolled on these platelet-decorated ULVWF strings, but not directly on endothelial cells, under high shear stresses of 20 and 40 dyn/cm(2) in a P-selectin dependent manner. These results suggest that the endothelial cell-bound ULVWF provide an ideal matrix to aggregate platelets and recruit leukocytes to endothelial cells under high shear stress. The observed phenomenon delineates a mechanism for leukocytes to be tethered to arterial endothelial cells under high shear, providing a potential link between inflammation and thrombosis. PMID- 15748249 TI - Laboratory monitoring of low-molecular-weight heparin therapy-part II. PMID- 15748250 TI - Laboratory monitoring of low-molecular-weight heparin therapy-part II. PMID- 15748251 TI - Laboratory monitoring of low-molecular-weight heparin therapy-part II. PMID- 15748252 TI - Hypercoagulability preceding cancer. Does hypercoagulability awaken dormant tumor cells in the host? PMID- 15748253 TI - Hypercoagulability preceding cancer. The oral cancer model. PMID- 15748254 TI - Hypercoagulability preceding cancer. Are anticoagulants also antineoplastic? PMID- 15748255 TI - Hypercoagulability preceding cancer. Venous thromboembolism and subsequent cancer risk. PMID- 15748256 TI - Hypercoagulability preceding cancer. The iron hypothesis. PMID- 15748257 TI - Mechanisms for acquired activated protein C resistance in cancer patients. PMID- 15748258 TI - Diagnosis of deep venous thrombosis and pulmonary embolus in patients with mucinous variant adenocarcinoma. PMID- 15748259 TI - Application of pulmonary embolism rule-out criteria to the UK Manchester Investigation of Pulmonary Embolism Diagnosis (MIOPED) study cohort. PMID- 15748260 TI - Homozygous type I Protein C deficiency in neonatal purpura fulminans with a novel frame-shift deletion of 10 base pairs in exon 8 of PROC gene. PMID- 15748261 TI - Treatment of patients with acquired inhibitors. PMID- 15748262 TI - The 46C-->T polymorphism in the factor XII gene (F12) and the risk of venous thrombosis. PMID- 15748263 TI - Should we screen Eastern Mediterranean sickle beta-thalassemia patients for inherited thrombophilia? PMID- 15748264 TI - A novel fibrinogen gamma chain mutation (gamma 239 Gln-->His) is the cause of dysfibrinogenemia Vicenza. PMID- 15748265 TI - Macrothrombocytopenia in velocardiofacial syndrome. PMID- 15748266 TI - The biphasic waveform in plasma: identifying the sepsis--coagulation crossroad? A rebuttal. PMID- 15748268 TI - Risk of bleeding with vitamin K antagonists compared with low-molecular-weight heparin after orthopedic surgery: a rebuttal. PMID- 15748270 TI - More on: Prevention of venous thromboembolism after orthopedic surgery: the EXPRESS study. PMID- 15748271 TI - More on: Properties of recombinant human thromboplastins that determine the International Sensitivity Index (ISI). PMID- 15748272 TI - More on: Measuring circulating cell-derived microparticles. PMID- 15748273 TI - More on: Fluvastatin inhibits upregulation of tissue factor expression by antiphospholipid antibodies on endothelial cells. PMID- 15748275 TI - Talking about ... PMID- 15748279 TI - Measuring body composition in overweight individuals by dual energy x-ray absorptiometry. AB - BACKGROUND: Dual energy x-ray absorptiometry (DXA) is widely used for body composition measurements in normal-weight and overweight/obese individuals. The limitations of bone densitometers have been frequently addressed. However, the possible errors in assessing body composition in overweight individuals due to incorrect positioning or limitations of DXA to accurately assess both bone mineral density and body composition in obese individuals have not received much attention and are the focus of this report. DISCUSSION: We discuss proper ways of measuring overweight individuals and point to some studies where that might not have been the case. It appears that currently, the most prudent approach to assess body composition of large individuals who cannot fit under the scanning area would be to estimate regional fat, namely the regions of thigh and/or abdomen. Additionally, using two-half body scans, although time consuming, may provide a relatively accurate measurement of total body fat, however, more studies using this technique are needed to validate it. SUMMARY: Researchers using bone densitometers for body composition measurements need to have an understanding of its limitations in overweight individuals and address them appropriately when interpreting their results. Studies on accuracy and precision in measurements of both bone and soft tissue composition in overweight individuals using available densitometers are needed. PMID- 15748278 TI - CyProQuant-PCR: a real time RT-PCR technique for profiling human cytokines, based on external RNA standards, readily automatable for clinical use. AB - BACKGROUND: Real-time PCR is becoming a common tool for detecting and quantifying expression profiling of selected genes. Cytokines mRNA quantification is widely used in immunological research to dissect the early steps of immune responses or pathophysiological pathways. It is also growing to be of clinical relevancy to immuno-monitoring and evaluation of the disease status of patients. The techniques currently used for "absolute quantification" of cytokine mRNA are based on a DNA standard curve and do not take into account the critical impact of RT efficiency. RESULTS: To overcome this pitfall, we designed a strategy using external RNA as standard in the RT-PCR. Use of synthetic RNA standards, by comparison with the corresponding DNA standard, showed significant variations in the yield of retro-transcription depending the target amplified and the experiment. We then developed primers to be used under one single experimental condition for the specific amplification of human IL-1beta, IL-4, IL-10, IL 12p40, IL-13, IL-15, IL-18, IFN-gamma, MIF, TGF-beta1 and TNF-alpha mRNA. We showed that the beta-2 microglobulin (beta2-MG) gene was suitable for data normalisation since the level of beta2-MG transcripts in naive PBMC varied less than 5 times between individuals and was not affected by LPS or PHA stimulation. The technique, we named CyProQuant-PCR (Cytokine Profiling Quantitative PCR) was validated using a kinetic measurement of cytokine transcripts under in vitro stimulation of human PBMC by lipopolysaccharide (LPS) or Staphylococcus aureus strain Cowan (SAC). Results obtained show that CyProQuant-PCR is powerful enough to precociously detect slight cytokine induction. Finally, having demonstrated the reproducibility of the method, it was applied to malaria patients and asymptomatic controls for the quantification of TGF-beta1 transcripts and showed an increased capacity of cells from malaria patients to accumulate TGF-beta1 mRNA in response to LPS. CONCLUSION: The real-time RT-PCR technique based on a RNA standard curve, CyProQuant-PCR, outlined here, allows for a genuine absolute quantification and a simultaneous analysis of a large panel of human cytokine mRNA. It represents a potent and attractive tool for immunomonitoring, lending itself readily to automation and with a high throughput. This opens the possibility of an easy and reliable cytokine profiling for clinical applications. PMID- 15748280 TI - Heterologous expression in Tritrichomonas foetus of functional Trichomonas vaginalis AP65 adhesin. AB - BACKGROUND: Trichomonosis, caused by Trichomonas vaginalis, is the number one, nonviral sexually transmitted infection that has adverse consequences for the health of women and children. The interaction of T. vaginalis with vaginal epithelial cells (VECs), a step preparatory to infection, is mediated in part by the prominent surface protein AP65. The bovine trichomonad, Tritrichomonas foetus, adheres poorly to human VECs. Thus, we established a transfection system for heterologous expression of the T. vaginalis AP65 in T. foetus, as an alternative approach to confirm adhesin function for this virulence factor. RESULTS: In this study, we show stable transfection and expression of the T. vaginalis ap65 gene in T. foetus from an episomal pBS-ap65-neo plasmid. Expression of the gene and protein was confirmed by RT-PCR and immunoblots, respectively. AP65 in transformed T. foetus bound to host cells. Specific mAbs revealed episomally-expressed AP65 targeted to the parasite surface and hydrogenosome organelles. Importantly, surface-expression of AP65 in T. foetus paralleled increased levels of adherence of transfected bovine trichomonads to human VECs. CONCLUSION: The T. vaginalis AP65 adhesin was stably expressed in T. foetus, and the data obtained using this heterologous system strongly supports the role of AP65 as a prominent adhesin for T. vaginalis. In addition, the heterologous expression in T. foetus of a T. vaginalis gene offers an important, new approach for confirming and characterizing virulence factors. PMID- 15748281 TI - Inhibition of breathing after surfactant depletion is achieved at a higher arterial PCO2 during ventilation with liquid than with gas. AB - BACKGROUND: Inhibition of phrenic nerve activity (PNA) can be achieved when alveolar ventilation is adequate and when stretching of lung tissue stimulates mechanoreceptors to inhibit inspiratory activity. During mechanical ventilation under different lung conditions, inhibition of PNA can provide a physiological setting at which ventilatory parameters can be compared and related to arterial blood gases and pH. OBJECTIVE: To study lung mechanics and gas exchange at inhibition of PNA during controlled gas ventilation (GV) and during partial liquid ventilation (PLV) before and after lung lavage. METHODS: Nine anaesthetised, mechanically ventilated young cats (age 3.8 +/- 0.5 months, weight 2.3 +/- 0.1 kg) (mean +/- SD) were studied with stepwise increases in peak inspiratory pressure (PIP) until total inhibition of PNA was attained before lavage (with GV) and after lavage (GV and PLV). Tidal volume (Vt), PIP, oesophageal pressure and arterial blood gases were measured at inhibition of PNA. One way repeated measures analysis of variance and Student Newman Keuls-tests were used for statistical analysis. RESULTS: During GV, inhibition of PNA occurred at lower PIP, transpulmonary pressure (Ptp) and Vt before than after lung lavage. After lavage, inhibition of inspiratory activity was achieved at the same PIP, Ptp and Vt during GV and PLV, but occurred at a higher PaCO2 during PLV. After lavage compliance at inhibition was almost the same during GV and PLV and resistance was lower during GV than during PLV. CONCLUSION: Inhibition of inspiratory activity occurs at a higher PaCO2 during PLV than during GV in cats with surfactant-depleted lungs. This could indicate that PLV induces better recruitment of mechanoreceptors than GV. PMID- 15748283 TI - Witch-hunt. AB - Beginning two years ago, the US Dept of Health and Human Services began "special reviews" of all current research grants that involved harm reduction, sex and drugs, and continues its ban on funding of needle exchange. With Bush's second term, the campaign was extended to all US funded international programs that dealt with these issues and populations. And, most recently, the US has again undertaken to dominate the discourse within international organizations charged with drug control and AIDS policies - especially those of the UN. But the international harm reduction and human rights community is fighting back in several important ways, including "An Open Letter to the delegates of the Forty eighth session of the Commission on Narcotic Drugs (CND) of the UN" prepared by a group of 334 well respected public health experts and human rights advocates, protesting U.S. pressure on the U.N. to withdraw its support from harm reduction. This editorial includes the letter and signatures as well as French, Spanish, and Russian versions of the letter as additional files. PMID- 15748282 TI - The effects of quercetin on SW480 human colon carcinoma cells: a proteomic study. AB - BACKGROUND: High fruit and vegetable intake is known to reduce the risk of colon cancer. To improve understanding of this phenomenon the action of different phytochemicals on colon cells has been examined. One such compound is quercetin that belongs to the group known as flavonoids. The purpose of this study was to determine the influence of quercetin on the proteome of the SW480 human colon adenocarcinoma cell line, specifically to identify proteins that could be the molecular targets of quercetin in its amelioration of the progression of colon cancer. To this end, two-dimensional gel electrophoresis and mass spectrometry were used to identify proteins that underwent a change in expression following treatment of the cells with 20 muM quercetin. This could elucidate how quercetin may reduce the progression of colon cancer. RESULTS: Quercetin treatment of the SW480 human colon cancer cells was found to result in the decreased expression of three proteins and the increased expression of one protein. The identified proteins with decreased expression were type II cytoskeletal 8 keratin and NADH dehydrogenase Fe-S protein 3. The other protein with decreased expression was not identified. The protein with increased expression belonged to the annexin family. CONCLUSION: Several proteins were determined to have altered expression following treatment with quercetin. Such changes in the levels of these particular proteins could underlie the chemo-protective action of quercetin towards colon cancer. PMID- 15748284 TI - On the methodological unification in electroencephalography. AB - BACKGROUND: This paper presents results of a pursuit of a repeatable and objective methodology of analysis of the electroencephalographic (EEG) time series. METHODS: Adaptive time-frequency approximations of EEG are discussed in the light of the available experimental and theoretical evidence, and applicability in various experimental and clinical setups. RESULTS: Four lemmas and three conjectures support the following conclusion. CONCLUSION: Adaptive time frequency approximations of signals unify most of the univariate computational approaches to EEG analysis, and offer compatibility with its traditional (visual) analysis, used in clinical applications. PMID- 15748285 TI - HLA class I and II genotype of the NCI-60 cell lines. AB - Sixty cancer cell lines have been extensively characterized and used by the National Cancer Institute's Developmental Therapeutics Program (NCI-60) since the early 90's as screening tools for anti-cancer drug development. An extensive database has been accumulated that could be used to select individual cells lines for specific experimental designs based on their global genetic and biological profile. However, information on the human leukocyte antigen (HLA) genotype of these cell lines is scant and mostly antiquated since it was derived from serological typing. We, therefore, re-typed the NCI-60 panel of cell lines by high-resolution sequence-based typing. This information may be used to: 1) identify and verify the identity of the same cell lines at various institutions; 2) check for possible contaminant cell lines in culture; 3) adopt individual cell lines for experiments in which knowledge of HLA molecule expression is relevant. Since genome-based typing does not guarantee actual surface protein expression, further characterization of relevant cell lines should be entertained to verify surface expression in experiments requiring correct antigen presentation. PMID- 15748286 TI - Arginase attenuates inhibitory nonadrenergic noncholinergic nerve-induced nitric oxide generation and airway smooth muscle relaxation. AB - BACKGROUND: Recent evidence suggests that endogenous arginase activity potentiates airway responsiveness to methacholine by attenuation of agonist induced nitric oxide (NO) production, presumably by competition with epithelial constitutive NO synthase for the common substrate, L-arginine. Using guinea pig tracheal open-ring preparations, we now investigated the involvement of arginase in the modulation of neuronal nitric oxide synthase (nNOS)-mediated relaxation induced by inhibitory nonadrenergic noncholinergic (iNANC) nerve stimulation. METHODS: Electrical field stimulation (EFS; 150 mA, 4 ms, 4 s, 0.5 - 16 Hz) induced relaxation was measured in tracheal preparations precontracted to 30% with histamine, in the presence of 1 microM atropine and 3 microM indomethacin. The contribution of NO to the EFS-induced relaxation was assessed by the nonselective NOS inhibitor L-NNA (0.1 mM), while the involvement of arginase activity in the regulation of EFS-induced NO production and relaxation was investigated by the effect of the specific arginase inhibitor nor-NOHA (10 microM). Furthermore, the role of substrate availability to nNOS in EFS-induced relaxation was measured in the presence of various concentrations of exogenous L arginine. RESULTS: EFS induced a frequency-dependent relaxation, ranging from 6.6 +/- 0.8% at 0.5 Hz to 74.6 +/- 1.2% at 16 Hz, which was inhibited with the NOS inhibitor L-NNA by 78.0 +/- 10.5% at 0.5 Hz to 26.7 +/- 7.7% at 8 Hz (P < 0.01 all). In contrast, the arginase inhibitor nor-NOHA increased EFS-induced relaxation by 3.3 +/- 1.2-fold at 0.5 Hz to 1.2 +/- 0.1-fold at 4 Hz (P < 0.05 all), which was reversed by L-NNA to the level of control airways in the presence of L-NNA (P < 0.01 all). Similar to nor-NOHA, exogenous L-arginine increased EFS induced airway relaxation (P < 0.05 all). CONCLUSION: The results indicate that endogenous arginase activity attenuates iNANC nerve-mediated airway relaxation by inhibition of NO generation, presumably by limiting L-arginine availability to nNOS. PMID- 15748287 TI - Are decisions using cost-utility analyses robust to choice of SF-36/SF-12 preference-based algorithm? AB - BACKGROUND: Cost utility analysis (CUA) using SF-36/SF-12 data has been facilitated by the development of several preference-based algorithms. The purpose of this study was to illustrate how decision-making could be affected by the choice of preference-based algorithms for the SF-36 and SF-12, and provide some guidance on selecting an appropriate algorithm. METHODS: Two sets of data were used: (1) a clinical trial of adult asthma patients; and (2) a longitudinal study of post-stroke patients. Incremental costs were assumed to be 2000 dollars per year over standard treatment, and QALY gains realized over a 1-year period. Ten published algorithms were identified, denoted by first author: Brazier (SF 36), Brazier (SF-12), Shmueli, Fryback, Lundberg, Nichol, Franks (3 algorithms), and Lawrence. Incremental cost-utility ratios (ICURs) for each algorithm, stated in dollars per quality-adjusted life year (dollars/QALY), were ranked and compared between datasets. RESULTS: In the asthma patients, estimated ICURs ranged from Lawrence's SF-12 algorithm at 30,769 dollars/QALY (95% CI: 26,316 to 36,697) to Brazier's SF-36 algorithm at 63,492 dollars/QALY (95% CI: 48,780 to 83,333). ICURs for the stroke cohort varied slightly more dramatically. The MEPS based algorithm by Franks et al. provided the lowest ICUR at 27,972 dollars/QALY (95% CI: 20,942 to 41,667). The Fryback and Shmueli algorithms provided ICURs that were greater than 50,000 dollars/QALY and did not have confidence intervals that overlapped with most of the other algorithms. The ICUR-based ranking of algorithms was strongly correlated between the asthma and stroke datasets (r = 0.60). CONCLUSION: SF-36/SF-12 preference-based algorithms produced a wide range of ICURs that could potentially lead to different reimbursement decisions. Brazier's SF-36 and SF-12 algorithms have a strong methodological and theoretical basis and tended to generate relatively higher ICUR estimates, considerations that support a preference for these algorithms over the alternatives. The "second generation" algorithms developed from scores mapped from other indirect preference-based measures tended to generate lower ICURs that would promote greater adoption of new technology. There remains a need for an SF-36/SF-12 preference-based algorithm based on the US general population that has strong theoretical and methodological foundations. PMID- 15748288 TI - Admission to hospital following head injury in England: incidence and socio economic associations. AB - BACKGROUND: Head injury in England is common. Evidence suggests that socio economic factors may cause variation in incidence, and this variation may affect planning for services to meet the needs of those who have sustained a head injury. METHODS: Socio-economic data were obtained from the UK Office for National Statistics and merged with Hospital Episodes Statistics obtained from the Department of Health. All patients admitted for head injury with ICD-10 codes S00.0-S09.9 during 2001-2 and 2002-3 were included and collated at the level of the extant Health Authorities (HA) for 2002, and Primary Care Trust (PCT) for 2003. Incidence was determined, and cluster analysis and multiple regression analysis were used to look at patterns and associations. RESULTS: 112,718 patients were admitted during 2001-2 giving a hospitalised incidence rate for England of 229 per 100,000. This rate varied across the English HA's ranging from 91-419 per 100,000. The rate remained unchanged for 2002-3 with a similar magnitude of variation across PCT's. Three clusters of HA's were identified from the 2001-2 data; those typical of London, those of the Shire counties, and those of Other Urban authorities. Socio-economic factors were found to account for a high proportion of the variance in incidence for 2001-2. The same pattern emerged for 2002-3 at the PCT level. The use of public transport for travel to work is associated with a decreased incidence and lifestyle indicators, such as the numbers of young unemployed, increase the incidence. CONCLUSION: Head injury incidence in England varies by a factor of 4.6 across HA's and PCT's. Planning head injury related services at the local level thus needs to be based on local incidence figures rather than regional or national estimates. Socio-economic factors are shown to be associated with admission, including travel to work patterns and lifestyle indicators, which suggests that incidence is amenable to policy initiatives at the macro level as well as preventive programmes targeted at key groups. PMID- 15748289 TI - Patterns of lung cancer mortality in 23 countries: application of the age-period cohort model. AB - BACKGROUND: Smoking habits do not seem to be the main explanation of the epidemiological characteristics of female lung cancer mortality in Asian countries. However, Asian countries are often excluded from studies of geographical differences in trends for lung cancer mortality. We thus examined lung cancer trends from 1971 to 1995 among men and women for 23 countries, including four in Asia. METHODS: International and national data were used to analyze lung cancer mortality from 1971 to 1995 in both sexes. Age-standardized mortality rates (ASMR) were analyzed in five consecutive five-year periods and for each five-year age group in the age range 30 to 79. The age-period-cohort (APC) model was used to estimate the period effect (adjusted for age and cohort effects) for mortality from lung cancer. RESULTS: The sex ratio of the ASMR for lung cancer was lower in Asian countries, while the sex ratio of smoking prevalence was higher in Asian countries. The mean values of the sex ratio of the ASMR from lung cancer in Taiwan, Hong Kong, Singapore, and Japan for the five 5 year period were 2.10, 2.39, 3.07, and 3.55, respectively. These values not only remained quite constant over each five-year period, but were also lower than seen in the western countries. The period effect, for lung cancer mortality as derived for the 23 countries from the APC model, could be classified into seven patterns. CONCLUSION: Period effects for both men and women in 23 countries, as derived using the APC model, could be classified into seven patterns. Four Asian countries have a relatively low sex ratio in lung cancer mortality and a relatively high sex ratio in smoking prevalence. Factors other than smoking might be important, especially for women in Asian countries. PMID- 15748290 TI - Local therapy with CpG motifs in a murine model of allergic airway inflammation in IFN-beta knock-out mice. AB - BACKGROUND: CpG oligodeoxynucleotides (CpG-ODN) are capable of inducing high amounts of type I IFNs with many immunomodulatory properties. Furthermore, type-I IFNs have been proposed to play a key role in mediating effects of CpG-ODN. The precise role of IFN-beta in the immunomodulatory effects of CpG-ODN is not known. OBJECTIVE: Here, we aimed to elucidate the role of IFN-beta in the anti-allergic effect of CpG motifs. METHODS: We assessed the immune response in OVA-primed/OVA challenged IFN-beta knockout (-/-) mice compared to wild type (WT) control, after intranasal and systemic treatment with synthetic CpG motifs. RESULTS: Vaccination with CpG-ODN reduced the number of cells in airways of OVA-sensitized WT but not IFN-beta-/- mice. Although airway eosinophilia was reduced in both treated groups, they were significantly higher in IFN-beta-/- mice. Other inflammatory cells, such as lymphocytes and macrophages were enhanced in airways by CpG treatment in IFN-beta-/- mice. The ratio of IFN-gamma/IL-4 cytokines in airways was significantly skewed to a Th1 response in WT compared to IFN-beta-/- group. In contrast, IL-4 and IgE were reduced with no differences between groups. Ag specific T-cell proliferation, Th1-cytokines such as IFN-gamma, IL-2 and also IL 12 were significantly lower in IFN-beta-/- mice. Surprisingly, we discovered that intranasal treatment of mice with CpG-ODN results in mild synovitis particularly in IFN-beta-/- mice. CONCLUSION: Our results indicate that induction of Th1 response by therapy with CpG-ODN is only slightly and partially dependent on IFN beta, while IFN-beta is not an absolute requirement for suppression of airway eosinophilia and IgE. Furthermore, our finding of mild synovitis is a warning for possible negative effects of CpG-ODN vaccination. PMID- 15748292 TI - Elevated creatine kinase activity in primary hepatocellular carcinoma. AB - BACKGROUND: Inconsistent findings have been reported on the occurrence and relevance of creatine kinase (CK) isoenzymes in mammalian liver cells. Part of this confusion might be due to induction of CK expression during metabolic and energetic stress. METHODS: The specific activities and isoenzyme patterns of CK and adenylate kinase (AdK) were analysed in pathological liver tissue of patients undergoing orthotopic liver transplantation. RESULTS: The brain-type, cytosolic BB-CK isoenzyme was detected in all liver specimens analysed. Conversely, CK activity was strongly increased and a mitochondrial CK (Mi-CK) isoenzyme was detected only in tissue samples of two primary hepatocellular carcinomas (HCCs). CONCLUSION: The findings do not support significant expression of CK in normal liver and most liver pathologies. Instead, many of the previous misconceptions in this field can be explained by interference from AdK isoenzymes. Moreover, the data suggest a possible interplay between p53 mutations, HCC, CK expression, and the growth-inhibitory effects of cyclocreatine in HCC. These results, if confirmed, could provide important hints at improved therapies and cures for HCC. PMID- 15748291 TI - Duration of streptozotocin-induced diabetes differentially affects p38-mitogen activated protein kinase (MAPK) phosphorylation in renal and vascular dysfunction. AB - BACKGROUND: In the present study we tested the hypothesis that progression of streptozotocin (STZ)-induced diabetes (14-days to 28-days) would produce renal and vascular dysfunction that correlate with altered p38- mitogen-activated protein kinase (p38-MAPK) phosphorylation in kidneys and thoracic aorta. METHODS: Male Sprague Dawley rats (350-400 g) were randomized into three groups: sham (N = 6), 14-days diabetic (N = 6) and 28-days diabetic rats (N = 6). Diabetes was induced using a single tail vein injection of STZ (60 mg/kg, I.V.) on the first day. Rats were monitored for 28 days and food, water intake and plasma glucose levels were noted. At both 14-days and 28-days post diabetes blood samples were collected and kidney cortex, medulla and aorta were harvested from each rat. RESULTS: The diabetic rats lost body weight at both 14-days (-10%) and 28-days ( 13%) more significantly as compared to sham (+10%) group. Glucose levels were significantly elevated in the diabetic rats at both 14-days and 28-days post-STZ administration. Renal dysfunction as evidenced by renal hypertrophy, increased plasma creatinine concentration and reduced renal blood flow was observed in 14 days and 28-days diabetes. Vascular dysfunction as evidenced by decreased carotid blood flow was observed in 14-days and 28-days diabetes. We observed an up regulation of inducible nitric oxide synthase (iNOS), prepro endothelin-1 (preproET-1) and phosphorylated p38-MAPK in thoracic aorta and kidney cortex but not in kidney medulla in 28-days diabetes group. CONCLUSION: The study provides evidence that diabetes produces vascular and renal dysfunction with a profound effect on signaling mechanisms at later stage of diabetes. PMID- 15748294 TI - Theodor Hellbrugge: 85 years of age - Ad multos transannos, sanos, fortunatos et beatos. AB - We honor Theo Hellbrugge's acclaimed endeavors in the rehabilitation, or rather the prehabilitation of handicapped children. So far, he has focused on obvious handicaps, and we trust that he will include concern for everybody's silent handicaps in the future by screening for abnormal variability inside the physiological range. Therein, we introduce cis- and trans-years, components of transdisciplinary spectra that are novel for biology and also in part for physics. These components have periods, respectively, shorter and longer than the calendar year, with a counterpart in magnetoperiodism. Transyears characterize indices of geomagnetic activity and the solar wind's speed and proton density. They are detected, alone or together with circannuals, in physiology as well as in pathology, as illustrated for sudden cardiac death and myocardial infarction, a finding calling for similar studies in sudden infant death syndrome (SIDS). As transyears can beat with circannuals, and depend on local factors, their systematic mapping in space and time by transdisciplinary chronomics may serve a better understanding of their putative influence upon the circadian system. Longitudinal monitoring of blood pressure and heart rate detects chronome alterations underlying cardiovascular disease risk, such as that of myocardial infarction and sudden cardiac death. The challenge is to intervene in a timely fashion, preferably at birth, an opportunity for pediatricians in Theo Hellbrugge's footsteps. PMID- 15748293 TI - Global transcriptome analysis of the C57BL/6J mouse testis by SAGE: evidence for nonrandom gene order. AB - BACKGROUND: We generated the gene expression profile of the total testis from the adult C57BL/6J male mice using serial analysis of gene expression (SAGE). Two high-quality SAGE libraries containing a total of 76 854 tags were constructed. An extensive bioinformatic analysis and comparison of SAGE transcriptomes of the total testis, testicular somatic cells and other mouse tissues was performed and the theory of male-biased gene accumulation on the X chromosome was tested. RESULTS: We sorted out 829 genes predominantly expressed from the germinal part and 944 genes from the somatic part of the testis. The genes preferentially and specifically expressed in total testis and testicular somatic cells were identified by comparing the testis SAGE transcriptomes to the available transcriptomes of seven non-testis tissues. We uncovered chromosomal clusters of adjacent genes with preferential expression in total testis and testicular somatic cells by a genome-wide search and found that the clusters encompassed a significantly higher number of genes than expected by chance. We observed a significant 3.2-fold enrichment of the proportion of X-linked genes specific for testicular somatic cells, while the proportions of X-linked genes specific for total testis and for other tissues were comparable. In contrast to the tissue specific genes, an under-representation of X-linked genes in the total testis transcriptome but not in the transcriptomes of testicular somatic cells and other tissues was detected. CONCLUSION: Our results provide new evidence in favor of the theory of male-biased genes accumulation on the X chromosome in testicular somatic cells and indicate the opposite action of the meiotic X-inactivation in testicular germ cells. PMID- 15748295 TI - A normalization strategy applied to HiCEP (an AFLP-based expression profiling) analysis: toward the strict alignment of valid fragments across electrophoretic patterns. AB - BACKGROUND: Gene expression analysis based on comparison of electrophoretic patterns is strongly dependent on the accuracy of DNA fragment sizing. The current normalization strategy based on molecular weight markers has limited accuracy because marker peaks are often masked by intense peaks nearby. Cumulative errors in fragment lengths cause problems in the alignment of same length fragments across different electropherograms, especially for small fragments (< 100 bp). For accurate comparison of electrophoretic patterns, further inspection and normalization of electrophoretic data after fragment sizing by conventional strategies is needed. RESULTS: Here we describe a method for the normalization of a set of time-course electrophoretic data to be compared. The method uses Gaussian curves fitted to the complex peak mixtures in each electropherogram. It searches for target ranges for which patterns are dissimilar to the other patterns (called "dissimilar ranges") and for references (a kind of mean or typical pattern) in the set of resultant approximate patterns. It then constructs the optimal normalized pattern whose correlation coefficient against the reference in the range achieves the highest value among various combinations of candidates. We applied the procedure to time-course electrophoretic data produced by HiCEP, an AFLP-based expression profiling method which can detect a slight expression change in DNA fragments. We obtained dissimilar ranges whose electrophoretic patterns were obviously different from the reference and as expected, most of the fragments in the detected ranges were short (< 100 bp). The normalized electrophoretic patterns also agreed well with reference patterns. CONCLUSION: The normalization strategy presented here demonstrates the importance of pre-processing before electrophoretic signal comparison, and we anticipate its usefulness especially for temporal expression analysis by the electrophoretic method. PMID- 15748296 TI - Clotting state after cardioversion of atrial fibrillation: a haemostasis index could detect the relationship with the arrhythmia duration. AB - BACKGROUND: Fibrin D-dimer levels have been advocated as an useful clinical marker of thrombogenesis. HYPOTHESIS: We hypothesized that i) there is a hyperclotting state after the return of atrial fibrillation to sinus rhythm, ii) the measurement of plasma D-Dimer levels might be a good screening tool of this clotting status, and iii) the duration of arrhythmia influences the haemostasis measured by plasma D-Dimer levels. METHODS: Forty-two patients with atrial fibrillation undergoing cardioversion were divided into two groups: in Group A (n = 24,14 male, 56 +/- 11 years) the duration of atrial fibrillation was 72 hours or more (142.7 +/- 103.8 hours), in Group B (n = 18, 10 male, 61 +/- 13 years) the duration of atrial fibrillation was less than 72 hours (25 +/- 16 hours). Plasma fibrin D-dimer levels were measured by enzyme immunoassay before, and 36 hours after, cardioversion. The change of plasma D-dimer levels 36 hours after cardioversion was calculated as delta-D-dimer. RESULTS: There were no significant differences in demographic, clinical, and echocardiographic data, and the success of cardioversion between the two groups. Compared to the control, the baseline D dimer levels were significantly higher in both groups. The delta D-dimer levels were significantly higher in Group A than in Group B (p < 0.005). Furthermore, plasma D-dimer levels 36 hours after cardioversion (r = 0.52, p = 0.0016) and delta-D-dimer levels (r = 0.73, p < 0.0001) showed significant correlations with the duration of atrial fibrillation. CONCLUSION: The longer duration of the atrial fibrillation episode could lead to a more prominent cardiovascular hyperclotting state after cardioversion, and the mean changes of plasma D-Dimer levels could be used as an useful clinical marker of the clotting state after atrial systole return. PMID- 15748297 TI - Cross-cultural adaptation of the VISA-A questionnaire, an index of clinical severity for patients with Achilles tendinopathy, with reliability, validity and structure evaluations. AB - BACKGROUND: Achilles tendinopathy is considered to be one of the most common overuse injuries in elite and recreational athletes and the recommended treatment varies. One factor that has been stressed in the literature is the lack of standardized outcome measures that can be used in all countries. One such standardized outcome measure is the Victorian Institute of Sports Assessment - Achilles (VISA-A) questionnaire, which is designed to evaluate the clinical severity for patients with Achilles tendinopathy. The purpose of this study was to cross-culturally adapt the VISA-A questionnaire to Swedish, and to perform reliability, validity and structure evaluations. METHODS: Cross-cultural adaptation was performed in several steps including translations, synthesis of translations, back translations, expert committee review and pre-testing. The final Swedish version, the VISA-A Swedish version (VISA-A-S) was tested for reliability on healthy individuals (n = 15), and patients (n = 22). Tests for internal consistency, validity and structure were performed on 51 patients. RESULTS: The VISA-A-S had good reliability for patients (r = 0.89, ICC = 0.89) and healthy individuals (r = 0.89-0.99, ICC = 0.88-0.99). The internal consistency was 0.77 (Cronbach's alpha). The mean [95% confidence interval] VISA A-S score in the 51 patients (50 [44-56]) was significantly lower than in the healthy individuals (96 [94-99]). The VISA-A-S score correlated significantly (Spearman's r = -0.68) with another tendon grading system. Criterion validity was considered good when comparing the scores of the Swedish version with the English version in both healthy individuals and patients. The factor analysis gave the factors pain/symptoms and physical activity CONCLUSION: The VISA-A-S questionnaire is a reliable and valid instrument and comparable to the original version. It measures two factors: pain/symptoms and physical activity, and can be used in both research and the clinical setting. PMID- 15748299 TI - Health care reform and job satisfaction of primary health care physicians in Lithuania. AB - BACKGROUND: The aim of this research paper is to study job satisfaction of physicians and general practitioners at primary health care institutions during the health care reform in Lithuania. METHODS: Self-administrated anonymous questionnaires were distributed to all physicians and general practitioners (N = 243, response rate - 78.6%), working at Kaunas primary health care level establishments, in October - December 2003. RESULTS: 15 men (7.9%) and 176 women (92.1%) participated in the research, among which 133 (69.6%) were GPs and 58 (30.4%) physicians. Respondents claimed to have chosen to become doctors, as other professions were of no interest to them. Total job satisfaction of the respondents was 4.74 point (on a 7 point scale). Besides 75.5% of the respondents said they would not recommend their children to choose a PHC level doctor's profession. The survey also showed that the respondents were most satisfied with the level of autonomy they get at work - 5.28, relationship with colleagues - 5.06, and management quality - 5.04, while compensation (2.09), social status (3.36), and workload (3.93) turned to be causing the highest dissatisfaction among the respondents. The strongest correlation (Spearmen's ratio) was observed between total job satisfaction and such factors as the level of autonomy - 0.566, workload - 0.452, and GP's social status - 0.458. CONCLUSION: Total job satisfaction of doctors working at primary health care establishments in Lithuania is relatively low, and compensation, social status, and workload are among the key factors that condition PHC doctors' dissatisfaction with their job. PMID- 15748298 TI - Quantitative inference of dynamic regulatory pathways via microarray data. AB - BACKGROUND: The cellular signaling pathway (network) is one of the main topics of organismic investigations. The intracellular interactions between genes in a signaling pathway are considered as the foundation of functional genomics. Thus, what genes and how much they influence each other through transcriptional binding or physical interactions are essential problems. Under the synchronous measures of gene expression via a microarray chip, an amount of dynamic information is embedded and remains to be discovered. Using a systematically dynamic modeling approach, we explore the causal relationship among genes in cellular signaling pathways from the system biology approach. RESULTS: In this study, a second-order dynamic model is developed to describe the regulatory mechanism of a target gene from the upstream causality point of view. From the expression profile and dynamic model of a target gene, we can estimate its upstream regulatory function. According to this upstream regulatory function, we would deduce the upstream regulatory genes with their regulatory abilities and activation delays, and then link up a regulatory pathway. Iteratively, these regulatory genes are considered as target genes to trace back their upstream regulatory genes. Then we could construct the regulatory pathway (or network) to the genome wide. In short, we can infer the genetic regulatory pathways from gene-expression profiles quantitatively, which can confirm some doubted paths or seek some unknown paths in a regulatory pathway (network). Finally, the proposed approach is validated by randomly reshuffling the time order of microarray data. CONCLUSION: We focus our algorithm on the inference of regulatory abilities of the identified causal genes, and how much delay before they regulate the downstream genes. With this information, a regulatory pathway would be built up using microarray data. In the present study, two signaling pathways, i.e. circadian regulatory pathway in Arabidopsis thaliana and metabolic shift pathway from fermentation to respiration in yeast Saccharomyces cerevisiae, are reconstructed using microarray data to evaluate the performance of our proposed method. In the circadian regulatory pathway, we identified mainly the interactions between the biological clock and the photoperiodic genes consistent with the known regulatory mechanisms. We also discovered the now less-known regulations between crytochrome and phytochrome. In the metabolic shift pathway, the casual relationship of enzymatic genes could be detected properly. PMID- 15748301 TI - Pertinent issues related to laparoscopic radical prostatectomy. AB - PURPOSE: We describe the critical steps of the laparoscopic radical prostatectomy (LRP) technique and discuss how they impact upon the pertinent issues regarding prostate cancer surgery: blood loss, potency and continence. RESULTS: A major advantage of LRP is the reduced operative blood loss. The precise placement of the dorsal vein complex stitch associated with the tamponading effect of the CO(2) pneumoperitoneum significantly decrease venous bleeding, which is the main source of blood loss during radical prostatectomy. At the Cleveland Clinic, the average blood loss of our first 100 patients was 322.5 ml, resulting in low transfusion rates. The continuous venous bleeding narrowed pelvic surgical field and poor visibility can adversely impact on nerve preservation during open radical prostatectomy. Laparoscopy, with its enhanced and magnified vision in a relatively bloodless field allows for excellent identification and handling of the neurovascular bundles. During open retropubic radical prostatectomy, the pubic bone may impair visibility and access to the urethral stump, and the surgeon must tie the knots relying on tactile sensation alone. Consequently, open prostatectomy is associated with a prolonged catheterization period of 2 - 3 weeks. Comparatively, during laparoscopic radical prostatectomy all sutures are meticulously placed and each is tied under complete visual control, resulting in a precise mucosa-to-mucosa approximation. CONCLUSION: The laparoscopic approach may represent a reliable less invasive alternative to the conventional open approach. Despite the encouraging preliminary anatomical and functional outcomes, prospective randomized comparative trials are required to critically evaluate the role of laparoscopy for this sophisticated and delicate operation. PMID- 15748302 TI - Quantification of tumor extension in prostate biopsies - importance in the identification of confined tumors. AB - OBJECTIVE: To assess the importance of quantifying the adenocarcinoma in prostate biopsies when determining the tumor's final stage in patients who undergo radical prostatectomy. To identify the best methodology for obtaining such data. PATIENTS AND METHODS: Prostate biopsies from 132 patients were examined, with determination of Gleason histological grade and tumor volume in number of involved fragments, tumor extent of the fragment mostly affected by the tumor and the total percentage of tumor in the specimen. Theses parameters were statistically correlated with the neoplasia's final stage following the evaluation of radical prostatectomy specimens. RESULTS: An average of 12 and a median of 14 biopsy fragments were evaluated per patient. In the univariate analysis the Gleason histological grade, the largest tumor extent in one fragment and the total percentage of tumor in the specimen were correlated with tumor stage of the surgical specimen. In the multivariate analysis, the Gleason histological grade and the total percentage of tumor were strongly correlated with the neoplasia's final stage. The risk of the tumor not being confined was 3 for Gleason 7 tumors and 10.6 for Gleason 8 tumors or above. In cases where the tumor involved more than 60% of the specimen, the risk of non-confined disease was 4.4 times. Among 19 patients with unfavorable histological parameters, Gleason > 7 and extension greater than 60% the tumor final stage was pT3 in 95%. CONCLUSION: When associated to the Gleason histological grade, tumor quantification in prostate biopsies is an important factor for determining organ confined disease, and among the methods, total percentage of tumor is the most informative one. Such data should be included in the pathological report and must be incorporated in future nomograms. PMID- 15748303 TI - The impact of prior prostatic surgery on urinary continence in patients undergoing orthotopic ileal neobladder. AB - OBJECTIVE: To establish if previous surgery for benign prostatic hyperplasia (transurethral resection of the prostate or open prostatectomy), age, and preservation of prostatic apex can influence postoperative urinary continence in patients submitted to radical cystectomy and orthotopic ileal neobladder. PATIENTS AND METHODS: We analyzed 62 patients with bladder cancer who were treated with radical cystectomy and orthotopic ileal neobladder between 1987 and 1998 and had been followed for at least 24 months. The average age and median follow up were 61 years and 53 months, respectively. Postoperative urinary continence was correlated with 3 factors: patient age, preservation of prostatic apex during surgical excision and prior prostatic surgery for benign disease. Patients were defined as incontinent when they had to use more than 1 protective pad at the daytime. RESULTS: The overall incidence of urinary incontinence was 12.9%(8 out of 62 patients). The only statistically significant factor that impacted upon urinary continence was previous prostatic surgery, with respectively 33% versus 7% rate of incontinence for patients previously operated on and for those without previous operation (p = 0.023 odds ratio = 6.5, 95% confidence interval). Preservation of prostatic apex did not reach difference, 12% versus 13%, for those with and without preservation, and age also did not influence the postoperative continence rate. CONCLUSIONS: Prior prostatic surgery for benign prostatic hyperplasia probably can increases the risk for postcystectomy incontinence and preservation of prostate apex did not affect the continence rate. This issue deserves to be considered by the surgeon and must be discussed previously with the patients when planning an orthotopic bladder replacement. PMID- 15748304 TI - The accuracy of (99m)Tc-DTPA scintigraphy in the evaluation of acute renal graft complications. AB - PURPOSE: Renal scintigraphy has been used for many years in the evaluation of renal transplants and can help in the diagnosis of graft complications, leading to prompt clinical management and preventing further deterioration of renal function. The purpose of this study was to evaluate the overall accuracy of renal scintigraphy with (99m)Tc-DTPA in the diagnosis of acute renal graft complications. MATERIALS AND METHODS: Seventy-six scintigraphic studies performed in 55 patients (ages ranging from 6 to 65 years), were reviewed. Scintigraphy results were compared to biopsies performed within 5 days of imaging. (99m)Tc DTPA study was performed within a mean time of 19 days after kidney transplants. Dynamic images were performed in the anterior position of the abdomen and pelvis every 2 seconds for 80 seconds (flow phase) and every 15 seconds for 30 minutes (functional phase), after an intravenous injection of 370 MBq (10 mCi) of (99m)Tc DTPA. RESULTS: The scintigraphic results were concordant with the biopsies in 86% of the cases studied. The sensitivities of renal scintigraphy for detection of acute tubular necrosis (ATN), acute rejection (AR) and cortical necrosis (CN) were 98%, 87% and 100%, respectively. Specificities and accuracies for detection of ATN, AR and CN were 89%, 86% and 100%, and 95%, 87% and 100%, respectively. CONCLUSION: Renal scintigraphy with (99m)Tc-DTPA showed a good overall accuracy in the detection of acute renal graft complications. It can be used as a reliable tool in the routine evaluation of these patients. PMID- 15748305 TI - Complete response of metastatic renal cancer with dendritic cell vaccine. AB - INTRODUCTION: We report a case of metastatic renal cell carcinoma that presented involution following therapy with dendritic cells. CASE REPORT: Male, 51-year old patient underwent left radical nephrectomy in September 1999 due to renal cell carcinoma, evolved with recurrence of the neoplasia in January 2002, confirmed by resection of the lesion. A vaccine therapy based on dendritic cells was then performed during 5 months (4 applications). After this period, there was occurrence of new lesions, whose resection revealed areas of necrosis and inflammatory infiltrate. DISCUSSION: The outcome of renal cell carcinoma is influenced by prognostic factors that confer more aggressive tumor characteristics. However, in cases of recurrence, the systemic therapy with dendritic cells-based vaccine can be associated with a better outcome with regression of disease. PMID- 15748306 TI - Port site recurrence after laparoscopic adrenalectomy for metastatic melanoma. AB - Port site metastasis after adrenal surgery is a rare entity. We describe the case of a 70-year-old man with a solitary adrenal metastasis from malignant melanoma, which was laparoscopically removed. Twelve months later, he presented a recurrence near one of the laparoscopic port sites. This lesion was surgically removed and after a 18-month follow-up, the patient presents no evidence of disease. PMID- 15748307 TI - Testicular chondrosarcoma. AB - A case of primary chondrosarcoma of the testis is reported. A 40-year-old man presented a painless swelling of the right testis that he has been observing for 3 years. Gross examination of the resected specimen showed an encapsulated, gray to tan colored, roughly rounded tumor. Histologically, the tumor revealed a well differentiated chondrosarcoma. PMID- 15748308 TI - Male perineal sling with autologous aponeurosis and bone fixation - description of a technical modification. AB - Post-prostatectomy urinary incontinence is an uncommon complication of adenomectomies, occurring in approximately 1% of cases and being more frequent following radical prostatectomies. There is a significant implication in the quality of life for these patients. The surgical techniques employed for its treatment are the implantation of an artificial sphincter, peri-urethral injections and suburethral slings. Considering the low efficacy of peri-urethral injections and the high cost of artificial sphincters, we present in this work a technical modification of the suburethral sling, whose preliminary results are satisfactory. The fundamental modification in this technique is due to the replacement of the synthetic material usually employed for making the sling for autologous tissue, constituted by an aponeurotic strip taken from the rectus muscle of abdomen. This modification aims to minimize risks of urethral erosion that, despite it was not described in this population due to the use of synthetic materials, is a possibility when facing the tension that is used over the bulbar urethra. In addition to such aspects the autologous aponeurosis does not have a cost except for a short prolongation of the surgical act. PMID- 15748309 TI - Ureteropelvic junction obstruction in children: two variants of the same congenital anomaly? AB - OBJECTIVE: To compare the characteristics of prenatally and postnatally diagnosed ureteropelvic junction obstruction (UPJO) in children. PATIENTS AND METHODS: We reviewed the records of 74 children who underwent pyeloplasty or nephrectomy for UPJO between 1995 and 2000. The patients were divided into 2 groups: prenatally and postnatally diagnosed UPJO. In each group, we compared age at surgery, gender, affected side, anteroposterior diameter (APD) of the renal pelvis, surgical findings, and renal function as determined by creatinine clearance. RESULTS: Of the 74 children, 44 (59.4%) had a prenatal diagnosis of UPJO and 30 (40.6%) had a postnatal diagnosis despite the fact that all had had a fetal ultrasonography. Median age at the time of surgery was 6.3 years (4 months to 16 years) for children with postnatal UPJO and 3.6 months (1 month to 4 years) for the prenatal group. Forty-three percent of the children in the postnatal group and 25% in the prenatal group were females. Clinical manifestations in children with postnatal UPJO included abdominal pain in 13 (43%) patients, pyelonephritis in 7 (23%), urinary tract infection in 5 (16.6%), and occasional findings upon ultrasound in 5 (16.6%). Excretory urography suggested obstruction in most children. The surgical findings included ureteral kinks due to adhesions in 93.3% of postnatally diagnosed UPJO cases and in 27.3% of prenatal cases (p < 0.01). A reduction in mean creatinine clearance of hydronephrotic kidneys was observed for both groups when compared to reference values for the respective ages, but this difference was not statistically significant. CONCLUSIONS: Postnatally diagnosed UPJO may be considered, at least in part, an entity different from prenatally detected obstruction due to its peculiar characteristics, i.e., postnatal UPJO more frequently affects females, manifests later in life with urinary infection or abdominal pain, and is frequently associated with ureteral kinking. PMID- 15748310 TI - Comparative study between intravenous urography and renal scintigraphy with DMSA for the diagnosis of renal scars in children with vesicoureteral reflux. AB - PURPOSE: To assess the value of intravenous urography (IVU) in detecting and grading the renal scar, comparing its results with those of scintigraphy with dimercaptosuccinic acid (DMSA). MATERIALS AND METHODS: The study included 43 children investigated by DMSA and IVU, who had vesicoureteral reflux diagnosed and classified through voiding cystourethrography. RESULTS: Among the kidneys with reflux, there was agreement between the results of DMSA and IVU concerning the presence and the absence of scars in 82.4% of the cases. Based on the results obtained, IVU would have a sensitivity of 66.6%, specificity of 94.4%; accuracy of 82.5%; positive predictive value (PPV) of 90% and negative predictive value (NPV) of 79%, when compared with DMSA results. Our data also confirm the close relation between the reflux grade and the presence of renal scar, since 75% of the kidneys with grade IV and V reflux presented scars. In relation to the grading of nephropathy, in 78% of patients the classification of the scar by both methods was identical. The highest disagreement was verified in the group with segmental scar on DMSA, where 41.6% of the kidneys were classified as normal on IVU. CONCLUSION: The data obtained confirm that the scintigraphy with DMSA is essential in the investigation of patients with renal scar, and cannot be replaced by IVU, due to its low sensitivity and lower ability of satisfactory grading. PMID- 15748331 TI - Efficacy of tamsulosin in the medical management of juxtavesical ureteral stones. PMID- 15748311 TI - Pubovaginal sling in the treatment of stress urinary incontinence for urethral hypermobility and intrinsic sphincteric deficiency. AB - PURPOSE: This study was undertaken to evaluate the use of pubovaginal sling for the treatment of female stress urinary incontinence in patients with intrinsic sphincteric deficiency and patients with urethral hypermobility. MATERIALS AND METHODS: Sixty-two patients aging 22 to 73 years-old (mean = 49.6) with a median parity of 4.1 (range 0 - 14) who underwent pubovaginal autologous fascial sling procedures for stress urinary incontinence from August/1999 to August/2002 were prospectively analyzed. Objective pre and postoperative urodynamic evaluation was performed in all cases. The patients were divided into 2 groups: thirty-nine patients (62.9%) with urethral hypermobility (Valsalva leak point pressure equal or superior to 60 cm of H(2)O) and twenty-three patients (37.1%) with intrinsic sphincteric insufficiency (Valsalva leak point pressure below 60 cm of H(2)O). RESULTS: The average follow-up period was 24.8 months, ranging from 3 to 38 months. Three patients (4.8%) had detrusor overactivity before the operation, and 36 patients (58.1%) had voiding dysfunction before surgery. The postoperative objective cure rate was 88.7% for stress urinary incontinence. The study also showed that 32.2% of the patients had voiding dysfunction and 11.3% had detrusor overactivity. The mean hospital stay was 3.1 days (range 2 - 4). No difference in the above parameters was noticed between patients with intrinsic sphincteric deficiency and those with urethral hypermobility. CONCLUSION: Construction of a pubovaginal sling is an effective technique for the relief of severe stress urinary incontinence, for both patients with urethral hipermobility and with intrinsic sphincteric deficiency, having a cure rate of 88.7%. The high frequency of postoperative voiding urgency was not related to the detrusor overactivity as evaluated by urodynamic studies. PMID- 15748332 TI - Urinary stone size: comparison of abdominal plain radiography and noncontrast CT measurements. PMID- 15748333 TI - International Brazil Journal Urology. PMID- 15748334 TI - Shock wave lithotripsy monotherapy for renal calculi. AB - Shock wave lithotripsy (ESWL) remains the most common treatment for renal calculi. In this article, recent literature pertaining to ESWL monotherapy of renal calculi was reviewed, with the goal of improving ESWL results through better case selection. When selecting the optimal surgical approach for a patient, multiple factors must be considered. Factors to consider include stone related factors (size, number, composition and location), renal anatomical factors, and patient-related factors. Each of these factors is presented in detail, with the discussion limit to non-staghorn renal calculi. Children, the elderly, patients with hypertension, and patients with impaired renal function, may be at increased risk of ESWL complications and adverse effects and care should be taken to limit the number and energy of shock waves applied in these special cases. Absolute contraindications to ESWL remain pregnancy, distal obstruction, untreated infection, and uncorrected coagulopathy. PMID- 15748335 TI - Endopyelotomy with the Acucise catheter. AB - OBJECTIVES: Determine the efficiency of the Acucise catheter in the treatment of ureteropelvic junction (UPJ) obstruction. MATERIALS AND METHODS: Between May 1997 and April 1999, 30 patients with primary or secondary ureteropelvic junction obstruction were treated using Acucise endopyelotomy catheter. The diagnosis was based on intravenous urography and diuretic renography. None of the patients underwent imaging studies for identification of blood vessels at the UPJ region. Twenty-one patients presented mild pyelocalyceal dilation and 9 had severe pyelocalyceal dilation. As for etiology, primary stricture was found in 24 patients and secondary stricture in 6. Three patients had UPJ obstruction associated with homolateral renal lithiasis. Retrograde Acucise endopyelotomy was performed in 27 patients; the antegrade access was used in 3 patients that had associated renal calculi. Incision of the stricture was performed laterally in all patients under fluoroscopic guidance. A 7F double-J catheter was placed immediately following the procedure, and removed after 6 weeks. Antegrade pyelotomy required a hospital stay of 3 days and retrograde pyelotomy required 1 day. RESULTS: Postoperative intravenous urography and diuretic renography demonstrated good results in 26 patients (86.6%). CONCLUSIONS: Acucise endopyelotomy represents a minimally invasive therapeutic option and can be used both in adults and in children 8 years old and above. Patients with UPJ obstruction and severe pyelocalyceal dilation had poorer results than those with mild pyelocalyceal dilation. PMID- 15748336 TI - Rigid ureteroscopy for diagnosis and treatment of ureteral calculi during pregnancy. AB - OBJECTIVE: To evaluate ureteroscopy as a treatment option for women presenting ureteral calculi during pregnancy. MATERIALS AND METHODS: Eighteen pregnant patients presenting renal colic and indication of surgical treatment for ureteral calculi were analyzed. Patients were 20 to 34 years old (medium = 28), and the gestation period ranged from 12 to 34 weeks (medium = 18). Lumbar pain was present in 14 patients, and 4 had diffuse abdominal pain. Four patients were febrile in the occasion of the examination. Thirteen patients presented microscopic hematuria, 8 leucocituria, and 4 positive urine culture. The stone was detected by ultrasonography (US) in 12 patients. Magnetic resonance imaging (MRI) was performed in 2 cases, and did not demonstrate calculi. The stone location was: 1 in the superior ureter (pregnancy of 15 weeks), 4 in the medium ureter (pregnancy of 12, 15, 18 and 20 weeks), 12 in the inferior ureter, and 1 was not determined. The surgical indication was difficult pain control, fever, and presence of uterine contractions. RESULTS: Double-J insertion, as single treatment, was possible in 4 patients and it was kept in place for up to 2 weeks after delivery. Among the patients submitted to ureteroscopy, the calculi retrieval was always possible, except in 1 case where the calculus was not located by US, MRI or ureteroscopy. In 2 patients, the ultrasonic lithotriptor was used and in 11 the stone was removed intact with a basket. There were no complications due to the procedure and all pregnancies were carried to full term. CONCLUSION: Rigid ureteroscopy for extraction of ureteral calculi during pregnancy is efficient and safe. PMID- 15748337 TI - CT-virtual endoscopy of the urinary tract. AB - OBJECTIVE: To demonstrate the various applications of CT-virtual endoscopy, for the assessment of urinary tract abnormalities. MATERIALS AND METHODS: Sixty-three patients were evaluated by CT-virtual endoscopy (49 CT-cystoscopy; 14 CT pyeloureteroscopy). CT-cystoscopy was obtained for follow-up of bladder tumor (n=21), radiologic suspicion (n=12) or radiologic evidence of urinary tract lesion (n=16). CT-pyeloureteroscopy was done due to neoplasm (n=5), calculi (n=3) and extrinsic compressions (n=3). RESULTS: In 49 patients submitted to CT cystoscopy, 27 tumors were detected intraoperatively (ranging 0.5-4.8cm). CT cystoscopy revealed 21 tumors (78%); all tumors larger than 0.6 cm were detected. Tumor within a bladder diverticulum was seen by CT-cystoscopy but not by endoscopy, in two patients. Useful additional information such as extension of tumors into the anterior portion of the bladder neck (n=2) and adequate characterization of bladder diverticulum in a child (n=1) was also obtained. CT pyeloureteroscopy detected 6 of 9 tumors (67%), and was useful in the differential diagnosis of pelvic/ureteral tumor versus calculi (n=8) and intrinsic versus extrinsic ureteral lesion (n=3). CONCLUSION: CT-virtual endoscopy is a useful procedure, particularly in the following situations: a)- Follow-up of bladder tumors; b)- Complimentary evaluation of areas of difficult approach by endoscopy; c)- Differential diagnosis of intrinsic versus extrinsic lesion of the renal pelvis and ureter. PMID- 15748338 TI - Periprostatic local anesthesia in transrectal ultrasound-guided prostate biopsy: is it possible to improve pain tolerance? AB - OBJECTIVE: Evaluate objectively pain tolerance in transrectal ultrasound-guided prostate biopsy (TRUS) using local periprostatic anesthesia applied intrarectally, compared to the conventional method. MATERIAL AND METHODS: Forty patients were submitted to TRUS-guided prostate biopsy due to clinical suspicious of neoplasia. Patients were randomized in 2 groups: group-I, with 20 patients submitted to local anesthesia by 4 periprostatic injection of 2.5 mL 1% lidocaine, without epinephrine, TRUS-guided; and group-II, with 20 controls, with no sedatives or analgesia. After biopsy, patients were questioned about pain intensity during the procedure, using a grading scale from 0 to 5, correlating numbers, colors, and pain intensity. Pain related to probe manipulation or biopsy punctures, acceptance of an eventual re-biopsy, side effects of the drug used, and later complications of the procedure were also evaluated. RESULTS: Both groups were consistent comparing PSA levels, and prostate volume. As for pain intensity, 18/20 patients had severe or intolerable pain on the group submitted to conventional biopsy, while for those submitted to periprostatic blockage this event occurred in 3/20 patients (Chi(2 ) =22.50; p<0.01). The most important pain component was manipulation of the transrectal probe in 28% of patients, and puncture itself in 72%. Acceptance of re-biopsy as a pain evaluation criterion occurred in only 45% of patients submitted to conventional biopsy, compared to 100% of those submitted to periprostatic anesthesia (Chi (2)=15.17; p<0.01). CONCLUSION: TRUS-guided prostate biopsy is a traumatic and painful experience, but the periprostatic blockage use is clearly associated with more tolerance and patient comfort during the exam. PMID- 15748339 TI - Localized prostatic cancer in patients submitted to renal transplant. AB - OBJECTIVE: An attempt is made to evaluate the incidence of prostate cancer in patients who have previously undergone a kidney transplant surgery and to determine the best therapeutic approach to this target group. MATERIAL AND METHODS: All kidney transplant male patients over 40 years of age were studied with respect to diseases unrelated to the transplants, which later affected them, mainly focusing on neoplastic disease and, more specifically, prostate cancer. RESULTS: Of 397 kidney-transplanted patients, 146 (37%) were males, at least 40 years old. Among the 10 of them (6.8%) who developed neoplastic diseases, there were two cases (1.4%) of prostatic cancer. Both were treated with a radical retropubic prostatectomy with no technical difficulty, in spite of the presence of a graft in one of the iliac fossa. CONCLUSIONS: Prostate cancer incidence in kidney transplant patients is still low (1.8%), but it will certainly heighten as transplants are performed in increasingly older people and as better immunosuppressive drugs are introduced to lengthen the survival of grafts and patients. Because these cancers are expected to be more aggressive as a consequence of continuous immunosuppression, early diagnosis is of critical importance, and those patients should be made aware of the need for frequent screening for prostate cancer. PMID- 15748340 TI - Laparoscopic removal of seminal vesicle cyst with ectopic ureteral insertion and renal remnant. AB - Seminal vesicle cysts associated with ectopic ureter and renal agenesis is a rare condition. We report on a 23-year-old man with a history of pelvic discomfort and post-coital testicular pain. The investigation disclosed a left seminal vesicle cyst, and an absent left kidney. The patient was successful submitted to resection of the left seminal vesicle, ureter, and dysplastic renal tissue altogether, through laparoscopic approach. Laparoscopy has shown to be an excellent treatment option for this rare condition. PMID- 15748341 TI - Rectourethral fistulas. AB - A recent review of the literature shows an increasing number of studies on this subject, most of which are retrospective and presented solely from a clinical point of view. Some of them are no more than case-reports. There is a lack of valid epidemiologic data about the incidence of rectourethral fistulas. According to the literature, these fistulas result from many different causes. Rectourethral fistulas may have a congenital or acquired origin. Acquired cases include inflammatory, neoplastic, or traumatic etiologies. The diagnostic algorithm is clear and very classical. The new diagnostic approach represents an MRI investigation of the male urethra. It is important to recognize this diversity of etiology because each type requires a different surgical strategy. The aim of the surgical approach is the closure of all types of fistulas. Spontaneous closure of the recto-urethral fistulas after double diversion or by means of a one-stage procedure is possible only in a few cases. In most cases, the treatment proceeds in three stages (double diversion-urinary and bowel, closure technique, undiversion). An endoscopic approach using biological sealants seems to be promising. Few urologists and general surgeons have attained wide experience in the management of rectourethral fistulas. No single procedure has been proven most effective or even universally applicable. PMID- 15748342 TI - Transperitoneal laparoscopic nephrectomy in children: surgical technique with 3 trocars. AB - INTRODUCTION: The first videolaparoscopic nephrectomy in children was performed in 1992, and since then, little experience, and small series of pediatric patients have been reported. The technique, described by Clayman and accepted worldwide, requires the insertion of 4 or 5 trocars. Introduction of trocars is an important cause of complication in videolaparoscopic surgery. The authors report laparoscopic transperitoneal nephrectomy in children using only 3 trocars, to minimize risk of vascular injury or visceral perforation. SURGICAL TECHNIQUE: The patient is placed in supine position with flank rotated at approximately 45(degrees). After pneumoperitoneum is established, the first trocar is introduced in umbilicus for the laparoscope. Under direct vision, the second trocar is placed at ipsilateral midclavicular line, and the third and last trocar in the epigastric region. Laparoscopic transperitoneal nephrectomy was performed in 3 children aged 7, 8, and 14 years old. Right nephrectomy was performed in 2 cases, and left nephrectomy in one. Mean operative time was 163 min (100 to 230 min), and no transfusion was necessary. Patients were discharged from hospital on day 2 to 4 after the procedure. There were no conversions to open surgery, and no intra or postoperative complications. COMMENTS: Every trocar and instrument introduction into the abdominal cavity presents an important risk of vascular injury or visceral perforation. The risk per patient is naturally increased with the number of trocars utilized. Injuries during laparoscopic procedures can theoretically damage every intra- or retroperitoneal organ. The majority of these lesions will need immediate or delayed open surgery, due to hematoma formation, postoperative bleeding, abscess, or peritonitis. Transperitoneal videolaparoscopic nephrectomy in children can be performed using only 3 trocars. The technique allows a better cosmetic result, and reduces the risk of trocar introduction injuries, like vascular and visceral lesions. PMID- 15748343 TI - Is the anterior vaginal wall sling a good alternative for intrinsic sphincteric insufficiency? AB - OBJECTIVES: We present our experience with the anterior vaginal wall sling, in the treatment of patients with stress urinary incontinence (SUI) due to urethral intrinsic sphincteric deficiency. MATERIAL AND METHODS: Forty-five women (mean age 53.4 years) with urodynamically proven intrinsic sphincteric deficiency (Valsalva Leak Point Pressure below 60cm H(2)O) were studied, prospectively. Coexisting bladder neck hypermobility was assessed using transperineal ultrasound. Patients with severe pelvic prolapse (grade 3 or 4) were excluded. Multivariable logistic regression was used to identify the variables that influenced the outcome (statistical significance was established for p<0.05). Follow-up ranged from 26 to 61 months (mean 40 months). RESULTS: Complete SUI cure was achieved in 14 women (31.1%) and 17 other women (37.8%) described SUI improvement and were satisfied with the outcome. Statistical analysis showed that factors such as age below 35 years (p=0.0251), and preoperative bladder neck hypermobility (p=0.0176), were strongly related to postoperative continence. CONCLUSIONS: We concluded that the vaginal wall sling has a high rate of failure in the treatment of patients with intrinsic sphincteric deficiency. This technique should not be proposed if hypermobility is not associated, especially in the case of elderly patients. PMID- 15748344 TI - Volume-weighted mean glomerular volume in spontaneously hypertensive rats treated with different doses of spironolactone. AB - OBJECTIVES: Ascertain alterations in volume-weighted glomerular volume in monotherapy of spontaneously hypertensive rats with different doses of spironolactone. MATERIAL AND METHODS: We have studied young adult male spontaneously hypertensive rats (SHR) 20 weeks old, divided in 4 groups of 5 animals each: Controls, spironolactone lower dose (5mg/kg/day), spironolactone average dose (10mg/kg/day), and spironolactone higher dose (30mg/kg/day) dissolved in drinking water (for 12 weeks). Serum albumin, blood urea nitrogen (BUN), and creatinine, and the volume-weighted mean glomerular volume (VWGV) were measured. RESULTS: Common progress of blood pressure (BP) in young adult SHR was greatly altered by spironolactone treatment, by either attenuation or reversion of BP increasing tendency. Biochemical evaluation of kidney function indicated normal levels for albumin and creatinine, and high levels for BUN in SHR in all groups. Thickened intrarenal arteries and venous stasis were observed in all groups. Treated SHR spironolactone average and higher doses glomeruli presented smaller and more regular profiles; glomerular hypertrophy with glomerular tuft gross disarray in controls and treated SHR spironolactone lower dose was observed. VWGV was significantly greater in control and treated SHR spironolactone lower dose than in treated SHR spironolactone average and higher doses. CONCLUSIONS: Monotherapy with spironolactone may affect glomerular size and shape in a dose-dependent way; spironolactone showed a significant effect in preserving VWGV, and may be used associated with other drugs in antihypertensive therapy to prevent secondary effects of hypertension in the kidney. PMID- 15748345 TI - Role of adjunctive medical therapy with nifedipine and deflazacort after extracorporeal shock wave lithotripsy of ureteral stones. PMID- 15748347 TI - Routine postoperative imaging is important after ureteroscopic stone manipulation. PMID- 15748346 TI - When is medical prophylaxis cost-effective for recurrent calcium stones? PMID- 15748348 TI - Ureteroscopic treatment of lower pole calculi: comparison of lithotripsy in situ and after displacement. PMID- 15748349 TI - Needle core length in sextant biopsy influences prostate cancer detection rate. PMID- 15748350 TI - Prostate cancer involving the bladder neck: recurrence-free survival and implications for AJCC staging modifications. PMID- 15748351 TI - Extraprostatic spread of clinically localized prostate cancer: factors predictive of pT3 tumor and of positive endorectal MR imaging examination results. PMID- 15748352 TI - Blunt renal trauma: minimally invasive management with microcatheter embolization - experience in nine patients. PMID- 15748353 TI - Sonic hedgehog signaling from the urethral epithelium controls external genital development. PMID- 15748354 TI - Predictive value of testicular histology in secretory azoospermic subgroups and clinical outcome after microinjection of fresh and frozen-thawed sperm and spermatids. PMID- 15748355 TI - Sperm aneuploidy rates in younger and older men. PMID- 15748356 TI - Microsurgical replantation of sexual organs in three patients. PMID- 15748357 TI - Vaginal reconstruction for ambiguous genitalia and congenital absence of the vagina: a 27-year experience. PMID- 15748358 TI - Multi-institutional experience with buccal mucosa onlay urethroplasty for bulbar urethral reconstruction. PMID- 15748359 TI - Critical evaluation of the problem of chronic urinary retention after orthotopic bladder substitution in women. PMID- 15748360 TI - Early results of pubovaginal sling lysis by midline sling incision. PMID- 15748362 TI - Increased urinary nitrite excretion in primary enuresis: effects of indomethacin treatment on urinary and serum osmolality and electrolytes, urinary volumes and nitrite excretion. PMID- 15748361 TI - Early hospital discharge for intravesical ureteroneocystostomy. PMID- 15748363 TI - International Brazil Journal Urology. PMID- 15748364 TI - Comparison of laparoscopic live donor nephrectomy versus the traditional open technique. AB - OBJECTIVES: Laparoscopic live donor nephrectomy (LDN) is a minimally invasive technique for kidney procurement that may decrease the donor disincentives. In addition, recent studies have demonstrated that LDN has equal graft and recipient survival when compared to the standard open approach. We report our experience with LDN and compare the results with the most recent open donor nephrectomy (ODN) group performed at our institutions. MATERIAL AND METHODS: The records of 70 consecutives left sided LDN performed between October 1998 and March 2001 were retrospectively reviewed and compared to 40 ODN performed between April 1996 and January 2000. RESULTS: Average blood loss (127 ml vs. 317 ml; p < 0.001), time to PO intake (25 hrs vs. 34.6 hrs; p < 0.001), and hospital stay (2.7 d vs. 4.2 d; p < 0.001) were statistically significant better for the LDN group when compared to ODN group. The average warm ischemia time in the LDN group was 138 seconds (range 55 - 360). The major complication rate in both laparoscopic (4 cases) and open (2 cases) donor groups was similar (5.7% and 5%, respectively). The average post operative day (POD) 90 recipient creatinine was similar for both groups (1.5+/ 0.9 vs. 1.5+/-0.8 ng/dL; p= 0.799). Similar rates of recipient ureteral complications occurred in the LDN and ODN groups: 1.4% (1 case) and 2.5% (1 case), respectively. Likewise, acute rejection was also similar at 22.8% (16 cases) and 27.5% (11 cases) in the LDN and ODN respectively. CONCLUSIONS: At our institutions, LDN was superior to ODN with regards to donor operative blood loss, time to PO intake, and length of hospital stay. In addition, similar complication rates, and 3-month recipient kidney function were demonstrated. PMID- 15748365 TI - Minimally invasive procedures for urethral incontinence: is there a role for laparoscopy? AB - This article focuses on the minimally invasive surgical approaches for the treatment of stress urinary incontinence (SUI). The role of laparoscopic suspension is reviewed and compared with other minimally invasive techniques, such as the pubovaginal sling procedure and injection of the urethral bulking agents. The role of laparoscopic Burch colposuspension remains ill defined in 2002. Once this minimally invasive technique is shown to duplicate the success rate of the open Burch procedure, it could be offered as a first-line therapy to patients with SUI. At this time, the pubovaginal sling (PVS) offers the best long term results with acceptable low complication rates of urinary retention, urgency, and sling erosion or infection. These complications are rarely seen with the laparoscopic repair but the incidence of bladder injuries is higher. The PVS operation can be performed as a salvage procedure, in obese patients, and concomitant with cystocele and rectocele repair whereas data for laparoscopy in these conditions are lacking. Until the long-term efficacy of the laparoscopic repair is clearly defined, offering it to patients as a minimally invasive therapy denies them of procedures with superior efficacy. PMID- 15748366 TI - Prevalence of high-grade prostatic intraepithelial neoplasia and its relationship to serum prostate specific antigen. AB - OBJECTIVE: We assessed the incidence of high-grade prostatic intraepithelial neoplasia (HGPIN) in prostates removed for bladder cancer, and evaluated its correlation with preoperative serum prostate specific antigen (PSA). MATERIALS AND METHODS: We reviewed the histology of the prostate from men who underwent cystoprostatectomy for bladder cancer without known prostate pathology prior to surgery. Preoperative serum PSA levels in patients with HGPIN were analyzed. RESULTS: 61 cystoprostatectomy specimens from May 1992 to April 1999 were reviewed. High-grade prostatic intraepithelial neoplasia (PIN) was found in 46/61 (75%) of the prostate specimens, including 21/21 (100%) patients with prostatic cancer, and 25/40 (63%) patients without prostatic cancer. The mean serum PSA in men with PIN without evidence of prostatic adenocarcinoma was 1.9 ng/mL (SD=2.026). CONCLUSION: The incidence of isolated high-grade PIN was 63%. The presence of high-grade PIN does not result in a significant elevation of serum PSA. PMID- 15748367 TI - Evaluating the efficiency of a combination of Pygeum africanum and stinging nettle (Urtica dioica) extracts in treating benign prostatic hyperplasia (BPH): double-blind, randomized, placebo controlled trial. AB - OBJECTIVES: In spite of its historical use, published data about phytotherapic products is characterized by the absence of well conducted studies, leading to conflictive and indefinite results about efficiency and safety of theses drugs. In that sense, we have analyzed the results of a combination of Pygeum africanum and stinging nettle (Urtica dioica) extracts in patients with benign prostatic hyperplasia (BPH), based in a double-blind, randomized, placebo-controlled protocol. MATERIALS AND METHODS: We have selected, according to inclusion and exclusion criteria, only patients with >or=50 years, presenting urinary symptoms assessed by the International Prostatic Symptoms Score (IPSS), with minimum score of 12, and Quality of Life (QoL) index of at least 3 points, rectal examination consistent with BPH, and maximum urinary flow rate (Q(max)) between 5 and 15 mL/s. Phytotherapic and placebo groups were formed by 27 and 22 patients, respectively. The major variables analyzed during the study were IPSS variation, Q(max), and side effects. Reduction of >or=30% and >or=50% in IPSS were the parameters used to define a clinically significant response (CSR). We have also analyzed >or=30% and >or=50% Q(max) increases. RESULTS: After six months of treatment we did not observe significant differences in clinical improvement potential between the phytotherapic combination and placebo groups. Percent IPSS drop of 21.6% in the phytotherapic group was similar to 19.7% obtained in the placebo group (p=0.928). Neither we observed any difference (p=0.530) for QoL improvement between phytotherapic (9.26%) and placebo (5.98%) groups. The alterations of Q(max) followed the trend line observed in clinical data, with no significant difference (p=0.463) in Q(max) increasing percent between phytotherapic (17.2%) and placebo (13.3%) groups. The CSR evaluation of clinical and urodynamic data was also similar between the groups. CONCLUSION: The combination of 25mg Pygeum africanum and 300mg stinging nettle extracts produced clinical and urodynamic effects similar to placebo in a group of HBP patients. PMID- 15748368 TI - Treatment of patients with superficial bladder cancer stratified by risk groups treated with lyophilized Moreau-Rio de Janeiro BCG strain. AB - INTRODUCTION: The Moreau-Rio de Janeiro BCG strain is considered the most effective to stimulate immunologic activity in mice. The objective of this prospective study was to evaluate BCG results for patients with superficial bladder cancer stratified by risk groups. MATERIAL AND METHODS: From April 1988 to May 2000, 100 patients were treated by transurethral resection for bladder tumor, followed by intravesical instillation of 40 mg BCG, with induction and maintenance cycles. Fisher exact test and Chi-square test, with 95% significance, were used to evaluate possible associations among variables. The Kaplan-Meier method was used to evaluate the disease-free interval and patients' survival, while log-rank test was used to compare the curves among the groups. RESULTS: The median follow-up was 69.3 months and varied from 10 to 153 months. Overall recurrence and progression rates were 55% and 13%, respectively. The medium time to recurrence was 9.4 months and to progression was 24.4 months. The cancer specific survive was 90%. Univariate analysis revealed that tumor recurrence was significantly associated with weekly BCG failure (p=0.011), multifocality (p=0.001), number of recurrences after primary therapy (p=0.001) and the need to Mitomycin C instillation (p=0.001). However, no variable was significantly associated with recurrence in multivariate analysis. There were significant associations, in univariate analysis, between disease progression and the following variables: tumor grade, weekly and 15-days BCG failure, both as first line and second line therapy, recurrence and need of Mitomycin C therapy. Independent variables to progression were 6.7 relative risk to weekly BCG failure, tumor grade and 15-days BCG (p= 0.08; CI=0.79-56.7), 2.4 (p= 0.11; CI=0.80-7.15) and 1.5 (p=0.23; CI=1.05-2.13), respectively. Patient stratification by risk groups were able to predict progression (p=0.045), but not recurrence (p=0.311). Disease progression rates were 3.2%, 12.2% e 25%, in low, intermediate and high risk groups, respectively. The BCG administration was well tolerated, and 21 patients (21%) didn't present any side effects. CONCLUSIONS: Intravesical instillation of BCG was overall well tolerated. Adjuvant BCG didn't decrease significantly recurrence rates, and 16% of the patients underwent alternative therapy with intravesical Mitomycin to prevent new recurrences. The risk group classification was able to select patients with high risk to progression. Tumor grade, BCG failure as first and second line therapies, were predictive factors of poor prognosis. BCG of Moreau-Rio de Janeiro strain was well tolerated, similar to other strains used in literature. PMID- 15748369 TI - Hydronephrosis secondary to a seminoma in abdominal cryptorchid testis. AB - The incidence of testicular cancer in cryptorchidism is 30 to 50 times higher than in general population. We present a case of hydronephrosis caused by a seminoma in an abdominal cryptorchid testis. A 22 years old male presented abdominal and right flank pain, and haematuria. Despite the fact that he could not feel right testis since he was 7 years old, he had never looked for treatment. A huge mass was palpable in hypogastric, and IVP and CT Scan demonstrated functional exclusion of the right kidney, and a solid tumor localized at the right iliac fossa, near the bladder wall. The patient was submitted to laparotomy with resection of the mass, and the histopathologic study showed classic seminoma. Abdominal testes have higher incidence of malignancy than those in the inguinal position. The orchiopexy is recommended for abdominal testis after 2 years old to facilitate the investigation and identification of testicular cancer. PMID- 15748370 TI - Dismembered laparoscopic pyeloplasty with antegrade placement of ureteral stent: simplification of the technique. AB - Laparoscopic pyeloplasty is an effective treatment option for ureteropelvic junction obstruction, with success rates superior to other minimally invasive approaches. We describe our technique of laparoscopic pyeloplasty with antegrade placement of ureteral stent with a laparoscopic-guided abdominal puncture. This technique decreases the use of fluoroscopy and also facilitates renal pelvis dissection and ureteropelvic anastomosis. PMID- 15748371 TI - Sexual rehabilitation after radical retropubic prostatectomy: new technique using ilio-inguinal nerve graft. AB - Sural nerve grafts have been used to repair the cavernous nerves and allow sexual potency recovery after radical prostatectomy. In order to overcome the drawbacks related to the harvesting of the sural nerve, the authors developed an original technique using ilio-inguinal nerve grafts to repair the cavernous innervation during radical prostatectomy. This technique used in a group of new patients proved its feasibility and absence of intra- and postoperative complications. Compared to sural nerve the use of ilio-inguinal graft has some advantages: 1)- Greater familiarity for urologists to harvest the ilio-inguinal nerve; 2)- Less time to acquire the graft; 3)- Greater postoperative comfort, avoiding inferior limb pain and edema, that delay patients' mobilization and recovery; 4)- Absence of anesthesia or paresthesia in the calcaneal area; 5)- Absence of sympathetic reflex dystrophy in inferior limbs. PMID- 15748372 TI - Urodynamic alterations in patients with HTLV-1 infection. AB - OBJECTIVES: The HTLV-1 is a retrovirus that causes several diseases, including tropical spastic paraparesis or HTLV-1 associated myelopathy (or TSP/HAM, as designated by the World Health Organization - WHO) described in 1985. In Brazil, the first cases were reported in 1989. In order to evaluate the urodynamic alterations of infected patients, 48 cases were studied: 26 TSP/HAM and 22 non TSP/HAM. MATERIAL AND METHOD: Evaluation was performed by testing, cystometry, abdominal pressure, differential pressure, detrusor leak point pressure, maximum flow pressure, and electromyography. RESULTS: 80.76% TSP/HAM patients showed hyperreflexic bladder, and 34.16% had detrusor-sphincter dyssinergia; 82.6% of this group had abnormal uroflow tests. Non-TSP/HAM patients had hyperreflexic bladders in 22.72% of the cases, and detrusor-sphincter dyssinergia was not assessed. For these patients, uroflow rate was normal in 70% of the cases. CONCLUSIONS: Patients infected by the HTLV-1, with or without myelopathy, present significant urodynamic abnormalities and must have a complete urologic and urodynamic evaluation. PMID- 15748373 TI - Tension-free vaginal tape (TVT): minimally invasive technique for stress urinary incontinence (SUI). AB - OBJECTIVE: The procedure o tension-free vaginal tape (TVT) was recently introduced in the therapeutic armamentarium of stress urinary incontinence (SUI). It is a pubovaginal sling applied in mid-urethra. The authors describe their experience with this technique. MATERIALS AND METHODS: During the period of May, 1999 to March, 2001, 110 patients (median age=53 years) were submitted to TVT implant for the treatment of stress urinary incontinence. Clinical exam and urodynamic studies were performed previously to surgery. All patients referred symptoms consistent with stress incontinence, and 35.7% complained of associated voiding urgency. Sixty-eight percent of the group reported previous continence procedure. Urethral hypermobility was diagnosed in 42% of patients (mean Valsalva leak point pressure under median effort of 110 cmH(2)O), and 58% had intrinsic sphincter deficiency (mean Valsalva leak point pressure under effort of 55 cmH(2)O). RESULTS: Mean patient follow-up was 18 months. Ninety-two percent of the patients were submitted to anesthetic blockade and 8% to local anesthesia. Mean length of the procedure was 30 minutes. Dystopy correction, when present, was performed in the same procedure. Mean hospital stay was 24 hours. There was bladder perforation in 13% of cases of TVT. There was no urethral or vaginal erosion. Twenty patients (18%) developed urgency symptoms during post-operative period, and 5 (4.5%) presented urge incontinence. During this follow-up period, 81% remained continent, 9% referred improvement of the symptoms compared to before the procedure and 10% were not satisfied. CONCLUSION: Our data allow us to conclude that TVT procedure is safe, rapid and promoting adequate levels of continence in mean term. PMID- 15748374 TI - Bladder augmentation in rabbits with anionic collagen membrane, with or without urotelial preservation. Cistometric and hystologic evaluation. AB - INTRODUCTION: The use of bowel segments to perform bladder augmentation is associated with several metabolic and surgical complications. A great variety of synthetic materials, biodegradable or not, have been tested. Collagen-based biomaterials have shown effectiveness for the regeneration and obtainment of a functional bladder. OBJECTIVE: Assess the functional and histological response of the rabbit bladder to anionic collagen membrane (ACM), either when it is anastomosed to the bladder or it is placed onto bladder after vesicomyectomy. MATERIALS AND METHODS: In 15 male rabbit a partial cystectomy was performed. After 4 weeks they were divided in 3 groups. Group 1 (G1) - bladder augmentation with ACM. Group 2 (G2) ACM is placed onto bladder after vesicomyectomy. Group 3 (G3) control group. Maximal bladder capacity (MBC) and weight were assessed with 4 (M1), 8 (M2) and 12 (M3) weeks after partial cystectomy. In M3 was performed the sacrifice and extraction of the bladder and kidneys for anatomopathologic study. RESULTS: There were neither bladder stones, nor implant extrusion in M3. There was a significant increase in MBC in G1 and G2 (p<0.05), but no statistical differences in G3 (p=0.35). There is no significant difference comparing G1 and G2. In M3, both groups have shown a bigger MBC than G3 (p<0.05). The microscopic assessment showed an inflammatory reaction in the bladder augmented, with urothelium preserved. CONCLUSIONS: The ACM was effective for the increase of MBC. The bladders with preservation of the urothelium have shown an extensive inflammatory process. PMID- 15748375 TI - Kidney damage and renal functional changes are minimized by waveform control that suppresses cavitation in shock wave lithotripsy. PMID- 15748376 TI - Safety and efficacy of holmium: YAG laser lithotripsy in patients with bleeding diatheses. PMID- 15748377 TI - Laparoscopic linear cutting stapler failure. PMID- 15748378 TI - One versus two proficient laparoscopic surgeons for laparoscopic live donor nephrectomy. PMID- 15748379 TI - Percent Gleason grade (4/5) as prognostic factor in prostate cancer diagnosed at transurethral resection. PMID- 15748380 TI - Phenotypic, molecular and ultrastructural studies of a novel low grade renal epithelial neoplasm possibly related to the loop of Henle. PMID- 15748382 TI - Testicular microlithiasis: prospective analysis of prevalence and associated tumor. PMID- 15748381 TI - Evaluation of sonographically guided percutaneous core biopsy of renal masses. PMID- 15748383 TI - Autologous penile corpora cavernosa replacement using tissue engineering techniques. PMID- 15748384 TI - Effects of various nitric oxide donating agents on the contractility and cyclic nucleotide turnover of human seminal vesicles in vitro. PMID- 15748385 TI - Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: a prospective study. PMID- 15748386 TI - Lack of standardization in performance of the semen analysis among laboratories in the United States. PMID- 15748387 TI - Phenotypic and functional characterization of in vivo tissue engineered smooth muscle from normal and pathological bladders. PMID- 15748388 TI - Tunica albuginea acellular matrix graft for penile reconstruction in the rabbit: a model for treating Peyronie's disease. PMID- 15748389 TI - Perioperative single dose instillation of epirubicin or interferon-alpha after transurethral resection for the prophylaxis of primary superficial bladder cancer recurrence: a prospective randomized multicenter study - Finnbladder III long term results. PMID- 15748390 TI - Factors explaining recurrence in patients undergoing chemoimmunotherapy regimens for frequently recurring superficial bladder carcinoma. PMID- 15748391 TI - Correlation of cystoscopy with histology of recurrent papillary tumors of the bladder. PMID- 15748393 TI - Outcome of the artificial urinary sphincter in female patients. PMID- 15748392 TI - Long-term results of Ingelman-Sundberg denervation procedures for urge incontinence refractory to medical therapy. PMID- 15748394 TI - 46,XY Intersex individuals: phenotypic and etiologic classification, knowledge of condition, and satisfaction with knowledge in adulthood. PMID- 15748395 TI - The single testis: paternity after presentation as unilateral cryptorchidism. PMID- 15748396 TI - International Brazil Journal Urology. PMID- 15748397 TI - Laparoscopic total and partial nephrectomy. AB - Laparoscopic radical nephrectomy has established its role as a standard of care for the management of renal neoplasms. Long term follow-up has demonstrated laparoscopic radical nephrectomy has shorter patient hospitalization and effective cancer control, with no significant difference in survival compared with open radical nephrectomy. For renal masses less than 4cm, partial nephrectomy is indicated for patients with a solitary kidney or who demonstrate impairment of contralateral renal function. The major technical issue for success of laparoscopic partial nephrectomy is bleeding control and several techniques have been developed to achieve better hemostatic control. Development of new laparoscopic techniques for partial nephrectomy can be divided into 2 categories: hilar control and warm ischemia vs. no hilar control. Development of a laparoscopic Satinsky clamp has achieved en bloc control of the renal hilum in order to allow cold knife excision of the mass, with laparoscopic repair of the collecting system, if needed. Combination of laparoscopic partial nephrectomy with ablative techniques has achieved successful excision of renal masses with adequate hemostasis without hilar clamping. Other techniques without hilar control have been investigated and included the use of a microwave tissue coagulator. In conclusion, laparoscopic radical nephrectomy for renal cell carcinoma has clearly demonstrated low morbidity and equivalent cancer control. The rates for local recurrences and metastatic spread are low and actuarial survival high. Furthermore, laparoscopic partial nephrectomy has demonstrated to be technically feasible, with low morbidity. With short term outcomes demonstrating laparoscopic partial nephrectomy as an efficacious procedure, the role of laparoscopic partial nephrectomy should continue to increase. PMID- 15748398 TI - Normal renal dimensions in a specific population. AB - INTRODUCTION: Renal dimensions (RD) are important for the diagnostic and the prognostic of nephropathies. MATERIALS AND METHODS: We selected 904 Brazilians subjects with normal excretory urographies, showing dense nephrogram at the 5th minute of the exam, serum creatinine < 1.3mg/dl, and absence of any disease that could modify RD. Length, width, and area of both kidneys were correlated with gender, age, height, and body weight. Five hundred and eighty one subjects were men (64.3%) and 323 were women (35.7%). Age ranged from 21 to 87 years old, body weight from 40 to 106kg (69.9+/-9.5 for men and 62.4+/-9.7 for women), and height from 1.37 to 1.94m (1.68+/-0.07 for men and 1.57+/-0.07 for women). RESULTS: There was an association (one-way Anova test) between length, width, and area, for each kidney and for both, with height (p<0.001), body weight (p<0.001), and gender (p<0.001). After adjustment for height (covariance analysis), both gender and body weight did not show influence on RD. Renal length and area reduced with aging (p<0.001), from the 7th decade compared to the others. Excluding these patients, height was the only variable to show association with RD, justifying data stratification by this variable. CONCLUSIONS: Renal length in this population showed that the normal patterns defined by other studies are inadequate for our population. Adjusting the data by height, gender, and body weight did not influence RD; however, the left kidney was bigger than the right kidney. Also, the influence of height was more pronounced below 1.66m. PMID- 15748399 TI - Predictive role of non-contrast spiral computerized tomography on spontaneous passage of ureteral stones. AB - PURPOSE: We assessed the role of non-contrast spiral computerized tomography (CT) for prediction of a favorable clinical outcome in patients with ureterolithiasis, presenting with acute flank pain. MATERIALS AND METHODS: Consecutive 185 patients having acute flank pain were prospectively evaluated with physical examination, urinalysis and hemogram, and non-contrast spiral CT. Size (greatest width in mm), location, perinephric fat stranding, the degree of hydronephrosis, tissue rim sign and perinephric fluid were assessed with spontaneously passed and unpassed stones. RESULTS: Urinary stone disease was investigated in 173 (93.5%) patients out of 185 (mean age = 41.1 years) by non-contrast spiral CT in whom ureterolithiasis was diagnosed in 96 (94 unilateral, 2 bilateral). Spontaneous passage was assumed in 79 patients with ureterolithiasis. Only 38 patients spontaneously passed ureteral stones with less than 7.4mm diameter. The greatest width difference was statistically significant between passed and unpassed group [(2.0-7.4mm; mean 4.37+/-1.63) vs. (4.0-10.0mm; mean 7.35+/-1.81), p<0.05]. CONCLUSIONS: Spiral CT seems to be a sensitive imaging modality for the detection of ureteral calculi. In addition, size and location of the ureteral stones and its effects on ureteral wall, as periureteral inflammation and edema, demonstrated by the rim sign, present an important predictive value on spontaneous passage of uretreral stones. PMID- 15748400 TI - Complications in radical cystectomy performed at a teaching hospital. AB - OBJECTIVES: Radical cystectomy with urinary diversion has been the standard treatment to infiltrative bladder neoplasms. We have analyzed a series of radical cystectomies performed by residents at a school hospital and compared the rates of complications in this series with those reported in literature. MATERIALS AND METHODS: During the period of December 1996 to December 2000, 59 surgeries were performed by 8 residents in our department, always oversaw by the same assistant. We have analyzed age, gender, pathological staging, type of diversion used, surgical time, need for blood transfusion and blood volume used during surgery, immediate complications (1 to 30 days), late complications, and follow-up time. RESULTS: There were 51 men and 8 women. Mean age was 61.3 years (28 to 89), and the majority of the group was over 60 years (60%). Mean follow-up time was 8.5 months (3 to 36). Camey II ileal neobladder was used in 36 (62%) cases, ureterosigmoidostomy in 8 (13.8%), Bricker in 12 (20.7%), and Mainz-Pouch II diversion in 2 (3.5%) cases. Blood transfusion was required in 25 (42%) cases. Immediate complications were observed in 19% of the patients, and late complications in 19% as well. CONCLUSION: When oversaw by an experienced surgeon, a resident gathers the conditions to perform a radical cystectomy, without significant increase in complication rates. PMID- 15748401 TI - Efficiency of short and long term antimicrobial therapy in transrectal ultrasound guided prostate biopsies. AB - OBJECTIVE: Prostate biopsy is a frequent diagnostic measure to detect prostatic conditions, including primarily prostate cancer. Its performance does not follow a pattern, mainly regarding preparation for the examination through antimicrobial prophylaxis. The aim of this study is to compare the efficiency of short and long term antimicrobial prophylaxis in transrectal ultrasound-guided prostate biopsies. MATERIALS AND METHODS: A clinical study was conducted with a total of 140 patients submitted to prostate biopsy with transrectal ultrasonographic control. Patients were randomly separated in two groups: Group 1 receiving norfloxacin 400mg single dose before the procedure and Group 2 receiving norfloxacin 400mg initiating before the procedure and then bid up to 6 doses. Efficiency control was determined by the incidence of urinary tract infection (UTI) and complications in both groups after statistical analysis. RESULTS: The incidence of minor complications in our study was 75%, consistent with data found in the literature. Among the patients with diagnosis of simple UTI, 23% belonged to Group 1 and 8% to Group 2 (p=0.08). Among the patients with complicated UTI, 37% belonged to Group 1 and none belonged to Group 2 (p=0.44). CONCLUSIONS: Several advances made transrectal ultrasound-guided prostate biopsies a useful and safe diagnostic tool in the workup of urologic patients. There is no optimal prophylactic preparation to the performance of this procedure. Long term antimicrobial prophylaxis presents a trend toward lower incidence of infectious complications. PMID- 15748402 TI - Bowel perforation during percutaneous renal surgery. AB - INTRODUCTION: We report the case of a 74 years old patient presenting bowel lesion in a percutaneous renal surgery to extract a 2.5cm diameter stone. CASE REPORT: The access to percutaneous nephrostolithotomy was performed between the 11th and 12th left ribs at the posterior axillary line. An abdominal transversal pad was placed and a guide was passed down the ureter with no difficulty. Surgical time was of approximately 40 minutes. Antegrade pielography performed at the end of the procedure was normal. In the first post-operative day, the patient presented severe pain and abdominal defense. An additional descendent pielography was performed gently pulling the nephrostomy catheter, thus contrasting the bowel. The patient underwent an exploratory laparotomy where lacerations in left colon external wall and a bowel transfixing lesion were found. Non-absorbable sutures in two layers were performed. The patient presented a good progress with no occurrences during post-operative period, and was discharged in one week. PMID- 15748403 TI - Textiloma nine years after nephrectomy. AB - INTRODUCTION: Foreign bodies after surgical procedures are not very reported in the literature. It is estimated to have 1 case for 1,300 operations, although in practice the frequency might probably be higher. CASE REPORT: A woman, 38 years, submitted to left nephroureterectomy for renal transplantation in 1993. During 9 years, she was asymptomatic, and then she presented intermittent left flank pain. Radiographic workup demonstrated a textiloma. DISCUSSION: In a review of the literature since 1950, solely 8 cases of textiloma in renal surgeries were reported, probably due to legal implications. PMID- 15748404 TI - Urolithiasis in children. AB - In general, the criteria for the treatment of urolithiasis in children are the same as those for adults. Today, extracorporeal shock wave lithotripsy (ESWL) is the method of choice in the treatment of most pediatric urinary stones. Stone free rates between 67% and 93% at short-term follow-up, and 57% to 92% at long term follow-up, have proven the efficacy of ESWL treatment in children. Nevertheless, the demand for auxiliary measures still remains. In order to achieve the most beneficial success rates under low complications, it is advisable to perform this type of ESWL in centers that claim the experience necessary for ESWL and endourological measures in children. PMID- 15748405 TI - Botulinum toxin treatment of urethral and bladder dysfunction. AB - There has been tremendous excitement with the use of botulinum toxin for the treatment of various urethral and bladder dysfunction over the past several years. Botulinum toxin is the most lethal naturally occurring toxin known to humankind. Why, then, would an urologist want to use this agent to poison the bladder or urethral sphincter? In this article, we will review the mechanisms underlying the effects of botulinum toxin treatment. We will discuss the current usage of this agent within the urologic community and will provide perspectives on future targets of botulinum toxin. PMID- 15748406 TI - Magnetic resonance imaging in the diagnosis of pelvic floor disorders. AB - Vaginal prolapse due to pelvic floor dysfunction occurs frequently in postmenopausal women. The disease usually involves all compartments of the vagina, so that isolated defects are uncommon. In advanced disease, it can be difficult to identify which organs are prolapsed, owing to the large bulge in vaginal area. Accurate diagnosis of pelvic floor defects, actual prolapsed organs, and presence of any coexisting abnormalities are essential to correctly plan surgical reconstruction and minimize the risk of recurrence. In this review, we discuss the existing imaging modalities available to evaluate pelvic prolapse, emphasizing the role of dynamic magnetic resonance imaging. PMID- 15748407 TI - Analysis of bladder histology and urodynamics in female rats submitted to bladder reconstruction with rectus abdominis muscle. AB - INTRODUCTION: Bladder histology and measures of maximal bladder pressure and maximal bladder volume during urinary loss in female rats submitted to bladder reconstruction with a myoperitoneal flap of rectus abdominis muscle were studied. MATERIALS AND METHODS: Fifty female adult Wistar rats were studied, separated in 5 groups: Group 1 (immediate): submitted to laparotomy, cystostomy, and cystometry; Group 2 (sham): submitted to laparotomy, anterior bladder wall incision, immediate suture and cystostomy; Group 3 (cystectomy): laparotomy, partial cystectomy, cystostomy; Group 4 (cystoplasty): midline laparotomy, partial cystectomy, augmentation with rectus abdominis muscle flap peritonized, cystostomy, total cystectomy, the rats were sacrificed and an histological study of the augmented bladder performed; Group 5 (cystostomy): midline laparotomy and cystostomy. Cystometry was always performed in post-operative day 7, excepting for Group 1. Assessment was comparative maximal bladder pressure and maximal bladder volume during urinary loss among the different groups. RESULTS: In cystoplasty group, mean maximal bladder pressure during urinary loss was lower than in immediate, sham, and cystostomy groups. It was also observed that maximal bladder volume during urinary loss presented mean and median values very close in each group, and cystectomy group showed much lower values. The group submitted to cystoplasty presented mean maximal bladder volume during urinary loss higher than all groups analyzed. Histological analysis of myoperitoneal flap augmented bladders showed partial and/or total epithelization in the muscular flap interspersed region with transitional cells, squamous metaplasia region and chronic inflammatory process. CONCLUSION: The use of peritonized rectus abdominis muscle flap to perform bladder augmentation was technically viable in the animals, showing urothelial epithelization in the muscular region of the flap, and a satisfactory gain of capacity and maintenance of low bladder pressures. PMID- 15748409 TI - Effect of low-carbohydrate high-protein diets on acid-base balance, stone-forming propensity, and calcium metabolism. PMID- 15748408 TI - Effect of potassium citrate therapy on stone recurrence and residual fragments after shockwave lithotripsy in lower caliceal calcium oxalate urolithiasis: a randomized controlled trial. PMID- 15748411 TI - Randomized prospective blinded study validating acquisition of ureteroscopy skills using computer based virtual reality endourological simulator. PMID- 15748412 TI - Adenocarcinoma of the prostate in young men: clinical and pathologic features. PMID- 15748410 TI - Is lower pole caliceal anatomy predictive of extracorporeal shock wave lithotripsy success for primary lower pole kidney stones? PMID- 15748413 TI - Why linear extent, not percent, of cancer should be used to measure cancer in needle biopsies. PMID- 15748415 TI - Renal angiomyolipoma: relationships between tumor size, aneurysm formation, and rupture. PMID- 15748414 TI - Prostate: high-frequency Doppler US imaging for cancer detection. PMID- 15748417 TI - Immunohistochemical localization of the retinoic acid receptors in human prostate. PMID- 15748416 TI - Effects of unilateral grade I testicular injury in rat. PMID- 15748418 TI - Laser welded vesicourethral anastomosis in an in vivo canine model: a pilot study. PMID- 15748420 TI - Newly diagnosed bladder cancer: the relationship of initial symptoms, degree of microhematuria and tumor marker status. PMID- 15748419 TI - After cystectomy, is it justified to perform a bladder replacement for patients with lymph node positive bladder cancer? PMID- 15748421 TI - Intravesical bacillus Calmette-Guerin reduces the risk of progression in patients with superficial bladder cancer: a meta-analysis of the published results of randomized clinical trials. PMID- 15748422 TI - Sexual function after using tension-free vaginal tape for the surgical treatment of genuine stress incontinence. PMID- 15748424 TI - Ultrasonography is unnecessary in evaluating boys with a nonpalpable testis. PMID- 15748423 TI - Does post-voiding residual volume get less as mobility improves in a rehabilitation ward for older adults? PMID- 15748425 TI - A ventral rotational skin flap to improve cosmesis and avoid chordee recurrence in epispadias repair. PMID- 15748426 TI - [Basic and clinical studies of the gene product-targeting therapy based on leukemogenesis--editorial]. AB - In the last twenty years, using all-trans retinoic acid (ATRA) as a differentiation inducer, Shanghai Institute of Hematology has achieved an important breakthrough in the treatment of acute promyelocytic leukemia (APL), which realized the theory of reversing phenotype of cells and provided a successful model of differentiation therapy in cancers. Our group first discovered in the world the variant chromosome translocation t(11;17)(q23;q21) of APL, and cloned the PML-RAR alpha, PLZF-RAR alpha and NPM-RAR alpha fusion genes corresponding to the characterized chromosome translocations t(15;17); t(11;17) and t(5;17) in APL. Moreover, establishment of transgenic mice model of APL proved their effects on leukemogenesis. The ability of ATRA to modify the recruitment of nuclear receptor co-repressor with PML-RAR alpha but not PLZF-RAR alpha caused by the variant chromosome translocation elucidated the therapeutic mechanism of ATRA from the molecular level and provides new insight into transcription-modulating therapy. Since 1994, our group has successfully applied arsenic trioxide (As(2)O(3)) in treating relapsed APL patients, with the complete remission rate of 70% - 80%. The molecular mechanism study revealed that As(2)O(3) exerts a dose-dependent dual effect on APL. Low-dose As(2)O(3) induced partial differentiation of APL cells, while the higher dose induced apoptosis. As(2)O(3) binds ubiquitin like SUMO-1 through the lysine 160 of PML, resulting in the degradation of PML-RAR alpha. Taken together, ATRA and As(2)O(3) target the transcription factor PML-RAR alpha, the former by retinoic acid receptor and the latter by PML sumolization, both induce PML-RAR alpha degradation and APL cells differentiation and apoptosis. Because of the different acting pathways, ATRA and As(2)O(3) have no cross-resistance and can be used as combination therapy. Clinical trial in newly diagnosed APL patients showed that ATRA/As(2)O(3) in combination yields a longer disease-free survival time. With the median survival of 18 months, none of the 20 cases in combination treatment relapsed, whereas 7 relapsed in 37 cases in mono-treatment. This is the best clinical effect achieved in treating adult acute leukemia to this day, possibly making APL the first adult curable leukemia. Based on the great success of the pathogenetic gene target therapy in APL, this strategy may extend to other leukemias. Combination of Gleevec and arsenic agents in treating chronic myeloid leukemia has already make a figure both in clinical and laboratory research, aiming at counteracting the abnormal tyrosine kinase activity of ABL and the degradating BCR-ABL fusion protein. In acute myeloid leukemia M(2b), using new target therapy degradating AML1-ETO fusion protein and reducing the abnormal tyrosine kinase activity of c kit will also lead to new therapeutic management in acute leukemias. PMID- 15748427 TI - [First report on assessment of the status of engraftment after allogeneic hematopoietic stem cell transplantation by using denaturing high-performance liquid chromatography]. AB - Monitoring engraftment of donor cells after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is supposed to be important for the early diagnosis of graft failure or relapse of malignancy. Several techniques have been reported for this purpose. PCR-based assays analyzing polymorphic short tandem repeats (STR) as markers are attractive because they are sensitive and can be performed rapidly. The intent of this study was to test a novel approach for assessment of donor engraftment using denaturing high-performance liquid chromatography (DHPLC) combined with STR-PCR. The feasibility of this assay and the accuracy of semi quantitative results were tested in-vitro by using serial DNA mixtures from unrelated individuals. The results showed that dilution experiments of the mock chimerism sample revealed a clear correlation between the percentage of donor or recipient DNA and the proportion of allele peak areas, with the limit of detection for a minor DNA percentage being 5%. Discrimination between donor and recipient was possible in all patients analyzed (n = 51) except for 5 patients whose pre-transplant samples were not available and identical twins in one case. STR results were the same as values obtained by capillary electrophoresis combined with fluorescence labeling multiply PCR. Results were also compared with data obtained with FISH analysis in a subgroup of patients receiving grafts from sex-mismatched donors or with PCR-detectable disease-specific gene products analysis. The results of the microsatellite analysis correlated well with the corresponding clinical findings. Full donor chimerism (FDC) were detected in all patients; decreasing values of donor chimerism were detected concomitantly with the appearance of relapse of disease in 3 patients. Samples from eight patients receiving HLA mismatched-haploidentical transplants from related donors together with cord blood transplants from unrelated donors were analyzed by this method. The results showed all 8 patients achieved FDC derived from related donors. It is concluded that this novel approach allows a rapid, sensitive, economical, auto mated and non-isotopic STR-PCR testing, thus provides a reliable alternative for assessment of the status of engraftment after allo-HSCT. PMID- 15748428 TI - [Peripheral blood T cell immuno-tolerance in PBSCT donors induced by rhG-CSF in vivo]. AB - The study was aimed to investigate the mechanism of T cell tolerance in human peripheral blood induced by rhG-CSF in vivo. Dendritic cell (DC) subsets, CD8(+)CD28(-) T suppressor cells and the expression of CD28 on T cells of peripheral blood before and after mobilization were analyzed by multicolor flow cytometry. The results showed that after mobilization by rhG-CSF in vivo, the relative counts of CD3(+)CD28(+) cells increased significantly (P < 0.01), and so did the CD8(+)CD28(+) cells (P < 0.01). The mean fluorescence intensity of CD28 expression on CD3(+) cells decreased greatly (P < 0.05), but there were no significant changes of the relative fluorescence intensity of CD28 overall expression on T cells (P > 0.05). The percentages of DC2 before mobilization were significantly lower as compared with normal bone marrow (P < 0.01). After using rhG-CSF, the DC2 count was significantly higher in the apheresis graft than in peripheral blood and bone marrow before mobilization (P < 0.01), while the DC1:DC2 ratios were lower (P < 0.01) and there was no significant difference of DC1 before and after mobilization (P > 0.05). The percentages of CD8(+)CD28(-) T suppressor cells increased significantly also after mobilization (P < 0.05). It is concluded that the higher numbers of DC2 and CD8(+)CD28(-) T suppressor cells in peripheral blood grafts may contribute to the ability of tolerance in peripheral blood T cells induced by rhG-CSF in vivo. PMID- 15748429 TI - [Effects of ligustrazine on expression of VCAM-1/VLA-4 in syngenic bone marrow transplantation of mice]. AB - To explore the effect of ligustrazine on the expression of adherent molecule VCAM 1/VLA-4 of bone marrow cells in syngenic bone marrow transplantation (BMT) mice, the mice were divided into 3 groups: normal group (which received no treatment), BMT control group and ligustrazine-treated groups. BMT mouse models were established. The BMT control group and the ligustrazine-treated group were orally administered 0.2 ml saline per mouse and 2 mg ligustrazine per mouse, respectively, twice a day. On the day 7, 14, 21, 28 after BMT, mice were respectively killed. Bone marrow nucleated cells were detected, and then the expression of VCAM-1/VLA-4 was assayed by immunohistochemistry, RT-PCR and flow cytometry analysis, respectively. The results showed that in ligustrazine-treated group, the accounts of bone marrow nucleated cells on the day 7, 14, 21, 28 after BMT were all higher than that in BMT control group. The expression level in the ligustrazine-treated group was significantly higher than that in the BMT control group (P < 0.05 or P < 0.01). It is concluded that ligustrazine can enhance VCAM 1/VLA-4 expression in bone marrow after syngenic bone marrow transplantation in mice, which may be related to the mechanisms underlying the ligustrazine accelerating hematopoietic reconstitution in allogenic bone marrow transplantation. PMID- 15748430 TI - [Detection of engraftment evidence after allogeneic hematopoietic stem cell transplantation by STR-PCR]. AB - The purpose of this study was to observe the chimera status of 15 patients received allogeneic hematopoietic stem cell transplantation by using STR-PCR method. DNA from peripheral blood or bone marrow of donors and recipients in different time were extracted, 5 STR loci with high polymorphism were amplified by PCR. The PCR products were analyzed by PAGE and silver staining. The results showed that 15 patients had different level of engraftment. 10 patients displayed complete chimerism, five patients showed mixed chimerism. 10 patients were keeping continuance of remission, 4 patients died and one patient relapsed but still alive. The decrease of donor DNA amounts in mixed chimerism foreshowed the early graft rejection or relapse. Incidence of I-II degree aGVHD high significantly correlated with mixed chimerism. It is concluded that STR-PCR is the sensitive and accurate method for analyzing the chimera status after allogeneic hematopoietic stem cell transplantation. Mixed chimerism is a prophetic role for leukemia relapse and chimera status guides the treatment. PMID- 15748431 TI - [Complications of successively double autologous hemopoietic stem cell transplants]. AB - In order to get clinical information about safety and feasibility of successively double autologous hemopoietic stem cell transplants (SD-AHSCT) in malignant hematological disease patients, the complications and hematological reconstitution after SD-AHSCT in 20 patients were analyzed retrospectively. 20 patients with hematologic malignancies received autologous peripheral blood stem/progenitor cell transplantation at the first transplant, and then were given autologous bone marrow transplantation as the second transplant at 4-10 months. The results showed that all the patients tolerated mobilization and collection of peripheral blood stem/progenitor cells as well as bone marrow collection. All the patients got enough hematological stem/progenitor cells for SD-AHSCT and achieved hematological reconstitution after SD-AHSCT. The speed of hematological reconstitution was positively correlated with the transfused quantity of hematological stem/progenitor cells (r = 0.968). The hematological reconstitution after the first autologous hemopoietic stem cell transplant (AHSCT) was earlier than that of the second (P < 0.05). There was no statistical difference between the first and the second AHSCT for the incidence of skin or mucous membrane bleeding (P > 0.05). No patients occurred massive hemorrhage during SD-AHSCT. The quantity of platelet transfusion in the second AHSCT was larger than that in the first AHSCT (P < 0.01). The incidence of oral ulcer in the first AHSCT was significantly higher than that in the second (P < 0.01). No statistical difference between the first and the second AHSCT was there in infectious sites, infectious pathogens and infection incidence (P > 0.10). All the complications were improved or cured, and no patients died of SD-AHSCT complications. In conclusion, SD-AHSCT is safe and feasible, and worthy to be further popularized. PMID- 15748432 TI - [Effects of HGF on GVHD and Th1/Th2-related cytokines in ALL mice after allo BMT]. AB - To observe the effects of hepatocyte growth factor (HGF) on graft-versus-host disease (GVHD) and Th1/Th2 related cytokines in mice with acute lymphoblastic leukemia (ALL) after allogenic bone marrow transplantation (allo-BMT), BALB/c mice were conditioned by total body irradiation with 11 Gy and then were transplanted with allogeneic bone marrow after establishing ALL model. BALB/c mice were divided into groups A and B. The mice of group A were injected subcutaneously with HGF from day 0 to 7 after allo-BMT, and the mice of group B were injected subcutaneously with PBS from day 0 to 7 after allo-BMT. The symptoms of GVHD and the GVHD pathological changes of liver and small intestine and skin were observed. The serum levels of both IFN-gamma and IL-4 were determined by ELISA. The results showed that the score of GVHD in group A was lower than that in group B (P < 0.05). The levels of IFN-gamma in both groups A and B were all higher than that in normal group (P < 0.05 and P < 0.001, respectively), However, the level of IFN-gamma in group A was lower than that in group B (P < 0.01). The levels of IL-4 in both group A and B were all lower than that in normal group (P < 0.05), but the level of IL-4 in group A was higher than that in group B (P < 0.05). It is concluded that HGF can alleviates the severity of GVHD, because of its balancing the Th1/Th2-related cytokines after allo-BMT. PMID- 15748433 TI - [ABO-incompatible nonmyeloablative allogeneic peripheral blood stem cell transplantation]. AB - To explore the effects of ABO incompatibility between recipient and donor on HLA matched nonmyeloablative allogeneic peripheral blood stem cell transplantation (NAST), a retrospective, cohort study was performed. Among 24 HLA-matched NAST, 15 were major ABO-incompatible and 9 minor. Control group included 24 HLA-matched NAST with ABO-compatible grafts. Nonmyeloablative conditioning regimens consisted of CTX, Ara-C and ATG. The patients were given cyclosporine A and mycophenolate mofetile for prophylaxis of acute GVHD. The ABO-incompatible patients received grafts depleted erythrocytes by hydroxyethyl starch (HES) sedimentation. The results showed that successful and stable engraftment was established in 23 patients. No recipient developed clinically immediate hemolysis during graft infusion, but 2 recipients experienced delayed hemolysis attributable to the ABO incompatibility. The median time of granulocyte counts >0.5 x 10(9)/L and platelet >30 x 10(9)/L was 11 and 14.9 days, respectively. In ABO major incompatible group, the onset of erythropoiesis after NAST was delayed. One out of 10 recipients with blood group "O" in this group developed pure red cell aplasia (PRCA), lasting 5 months. The acute GVHD occurred in 7 out of the 24 patients. The chronic GVHD occurred in 5 of 21 cases. Relapse was observed in 2 patients with acute leukemia. The actuarial probability of disease-free survival at 2 years was 63.3%. In conclusion, ABO-incompatible grafts for NAST have no adverse effect on engraftment, recovery of platelets, incidence of GVHD, relapse rate or survival. ABO-incompatible NAST is fairly safe if there is indication, however, the onset of erythropoiesis is delayed when major ABO mismatched. PMID- 15748434 TI - [Effect of human bone marrow mesenchymal stem cell on allogeneic T lymphocyte phenotype in vitro]. AB - The purpose was to study the effect of mesenchymal stem cell (MSC) on immune function, and to explore the new strategy to prevent graft versus host disease (GVHD) and host versus graft reaction (HVGR) in allogeneic bone marrow transplantation. MSCs from human bone marrow cells were isolated and cultured. The purity of MSCs were identified with the spindle-fibroblastic morphological characterization by microphotography, and the phenotypes were tested by flow cytometry. MSCs were planted in 6-well plates (8 x 10(4)/well for group A, 4 x 10(4)/well for group B and 2 x 10(4)/well for group C), and cocultured for 7 days with T cell isolated from peripheral blood. Peripheral blood T cells non cocultured with MSC acted as the control group. CD3, CD4, CD8, and CD25 expressed on T cells were analyzed by flow cytometry after coculture with MSCs for 0, 24, 72 hours and 7 days respectively. The results showed that CD4(+)CD25(+) T cells and CD8(+) T cells of allogeneic T lymphocytes cocultured with bone marrow MSCs (group A and group B) increased obviously as compared with control group (T lymphocytes uncocultured with MSCs), and there were no difference between groups A and B. CD3, CD4, CD8 and CD25 expressed on T cells had no significant difference between experimental groups and control group. The result suggested that CD4(+)CD25(+)-regulatory T cells and CD8(+) T cells were increased apparently when cocultured with allogeneic MSCs. It is concluded that MSCs pretreatment may be useful in the prevention of GVHD and HVGR in allogeneic bone marrow transplantation and provides a new strategy to induce transplantation tolerance. PMID- 15748435 TI - [Osteoblasts derived from mesenchymal stem cells harbor immunoregulatory effect]. AB - In an attempt to study the immunoregulatory effect of osteoblasts derived from mesenchymal stem cells (MSC), MSC was induced to differentiate into osteoblasts for one week. The growth pattern and the phenotype were evaluated by MTT and flow cytometry respectively. The immunoregulatory effect was tested by the inhibitory effect on T cell proliferation. The result showed that during the differentiation cells grew fast and there was no significant change in the phenotypes but keeping CD73, CD105, CD44, CD29 positive and CD34, CD45, HLA-DR, CD86 negative. Osteocyte derived from MSC also showed immunosuppressive effect on T cell proliferation in adose-dependent manner. It is concluded that osteoblasts derived from MSC also harbored immunoregulatory effect. PMID- 15748436 TI - [Comparative study on various subpopulations in mesenchymal stem cells of adult bone marrow]. AB - To explore the difference of biological characteristics between two subpopulations of adult bone marrow mesenchymal stem cells (MSC), this study was designed to observe the morphological feature and immunophenotype of the adult MSC in the ex vivo culture, the mononuclear cells isolated from normal adult bone marrow were cultured in DMEM with 10% fetal bovine serum. Cell morphology, immunophenotype and cell cycle of two different subgroups were investigated. Cells from 80% confluence were passed through a 10 microm filter, then the fillered cells were cultured in the semisolid methylcellulose medium. The results showed that (1) two different subpopulations were observed in the ex vivo culture. The fibro-like cell was called mature MSC (mMSC) and the smaller round cell was defined rapidly as MSC self-renewing cells (RS cells); (2) the average proportion of cells in G(0)/G(1) of RS cells was approximately 99%, but that of mMSCs was 90%; (3) both of the two populations were negative on the lineage committed antigen (such as CD34, CD45, CD3, CD19, CD33, HLA-DR, CD38), while positive on the expression of CD90, CD105, C166, CD29, CD44, CD49e, CD54, CD13. However, the expression of these antigens on RS cells was weaker than that on mMSC, but CD117 and KDR were higher expressed when compared with the mMSC; (4) after 4 to 5 week semisolid culture, no hematopoietic progenitor cell colonies were observed. It is concluded that adult MSCs are heterogeneous in that distinct morphological populations exist. The RS cells appear to be the more primitive with greater potential for self-renewal and multilineage differentiation. PMID- 15748438 TI - [Expression and its significance of TRAIL and its receptors in cells of patients with acute myeloid leukemia]. AB - This study was aimed to detect the expression of TNF related apoptosis-inducing ligand (TRAIL) and its receptors on acute myeloid leukemic (AML) cells, and explore its possible role in leukemia therapy. RT-PCR and flow cytometry were used to detect the expression of TRAIL and its receptors on AML cells of 39 cases (patient group), AML cells of 18 cases with complete remission (CR group) and BMMNC or PBMNC of 21 normal persons (control group). The results showed that (1) TRAIL, DR4 and DR5 were highly expressed in both patient group and CR group, while the DcR1 and DcR2 were poorly expressed. (2) The level of DR5 expression in CR group was higher than that in patient group. (3) The level of DR5 was higher than DR4 in both patient group and CR group. (4) TRAIL and its receptors were expressed similarly in different subtypes of AML. In conclusion, there are differences between the expressions of TRAIL and its receptors in AML cells. DR5 may play an important role in TRAIL-inducing apoptosis of AML cells. PMID- 15748437 TI - [Observation on the biological behavior of human umbilical cord blood adherent cells]. AB - To study the possibility of separation and culture of human umbilical cord blood adherent cell (HUCBAC), the umbilical cord blood CD34(+) cells were cultured in Dexter system in order to evaluate and observe the biological behavior of adherent cells in vitro. The results showed that all cells were cultured with Dexter system. By day 9-14 (at a median of 11.2 days), adherent cell colonies formed and reached their maximum at 15-22 days (mean 19.6 days), by day 28, all adherent cells spread over the bottom of Petri dish. By means of light microscopy, these cells were found to differentiate into three kinds of cells in culture of 28 days: fibroblast-liked cell, macrophage liked cell and small-round cells. The ratio of these three kinds of cells was 56.8%, 38%, 5.5% respectively. Cytochemistry assay revealed that the positive rate reached 100% in NSE stain and PAS stain; the adherent cell by ALP stain were shown 35% positive, but in POX stain the result was negative. Immunohistochemistry stain revealed that the positive rate of cord adherent cells for CD106, CD29, CD44, CD45, CD50, Fn, Ln, collagen IV etc reached 96%, 93%, 98%, 68%, 72%, 92%, 74%, 83% respectively. It is concluded there are hematopoietic adherent precursors in cord blood CD34(+) cells and the HUCBAC shows some biological behavior of hematopoietic stromal cells. PMID- 15748439 TI - [Expressions of transcription factor GATA-1 and GATA-2 genes in bone marrow stromal cells from patients with leukemia]. AB - In order to investigate expressions of transcription factor GATA-1 and GATA-2 genes in the bone marrow stromal cells (BMSCs) from patients with leukemia or normal controls, bone marrow stromal cells from 34 normal cases and 42 cases with leukemia were cultured long-term in vitro. Nonadherent cells (bone marrow hematopoietic cells) and amplified adherent cells (BMSC) were collected separately. Expressions of GATA-1 and GATA-2 genes were analyzed by using RT-PCR ELISA; the semi-quantitative expression levels of GATA genes in the BMSCs from patients with leukemia were compared with normal controls. The results showed that expressions of GATA-1 and GATA-2 genes could be detected in the BMSCs and the bone marrow hematopoietic cells from both normal controls and the cases of leukemia. The expression ratio of GATA-1 in the BMSCs from acute lymphocytic leukemia (ALL) (85.7%) was similar to the normal controls (88.2%), whereas the expression ratios in BMSCs from acute myelocytic leukemia (AML) (55.6%) and chronic myelocytic leukemia (CML) (41.2%) were significant lower than the normal controls (P < 0.05). The rank of expression level of GATA-1 gene in the BMSCs was "ALL>AML>normal>CML". There was no difference in the expression level of GATA-2 gene within the BMSCs from normal controls and patients with leukemia. The ranks of expression levels of GATA-1 and GATA-2 genes in bone marrow hematopoietic cells were "AML>normal>ALL>CML" and "AML>CML>ALL>normal". The dominant expression of GATA-2 gene was found in the BMSCs from AML, CML or normal controls. It is inferred that the expressions of GATA-1 and GATA-2 genes in the BMSCs of normal controls and patients with leukemia may influence the regulation of hematopoiesis in the bone marrow stroma and it is worthy of further study to explore their roles in pathogenesis and development of leukemia. PMID- 15748440 TI - [Clinical significance of detection of AML1/ETO fusion transcripts in childhood AML using real-time quantitative reverse transcription polymerase chain reaction]. AB - This study was aimed to investigate the clinical value of quantification of AML1/ETO fusion transcripts using real-time reverse transcription PCR. Fourteen AML1/ETO positive children out of 52 AML children were selected. A serial dilution of AML1/ETO plasmid was used as a template for the AML1/ETO real-time PCR. AML1/ETO was quantified according to the expression of the GAPDH housekeeping gene at new diagnosis and during/after chemotherapy and transplantation. SPSS statistics was used to analyze the data. The results showed that the ratio of AML1/ETO: GAPDH expression level at new diagnosis varied in the range 0.219-2.080 (median 0.648) among the patients, without relevance with percentage of blasts. The detection sensitivity was up to the dilution of 1:10(5). Six patients showed a slight decline of AML1/ETO (higher than 5 x 10( 2)) at 1 month, three of whom relapsed in the early stage and one later. Five patients had a higher level than 5 x 10(-3) at 3 months, three of whom relapsed. Four patients with always a higher level than 5 x 10(-3) all relapsed in early stage. After six months, four out of them with constant low-level expression (10( 4) - 10(-6)) were in continuous complete hematological remission (CCR). In another patient, a rapid rise of AML1/ETO transcripts could be detected at CR stage and he relapsed 5 months later. The AML1/ETO gene expression leveling off by 10(-5) - 10(-6) could be detected in 3 patients at their complete remission after 9 months. It is concluded that real-time RT-PCR is a suitable approach for quantifying AML1/ETO transcripts in monitoring of AML patients with t(8;21) during/after chemotherapy and provides data of diagnostic relevance. PMID- 15748441 TI - [RT-PCR detecting NUP98-HOX fusion gene in leukemia]. AB - To investigate whether there are NUP98-HOXA, NUP98-HOXB, NUP98-HOXC, NUP98-HOXD fusion genes in leukemia patients in Xinjiang, cellular total RNA was extracted from the bone marrow mononuclear cells, the formaldehyde-agarose gel electrophoresis was used to judge whether RNA was intact, the 17 RT-PCR primers were designed to amplify the predicted fusion junctions and 412 bp GAPDH was used as an internal control, NUP98-HOXA fusion genes were amplified by nested-PCR following reverse transcription. One-step PCR was performed to amplify the other predicted fusion genes. The results showed that RNA was proved to be intact and expression of GAPDH was found in every sample. However, no predicted fusion transcripts were detected in leukemia patients. In conclusion, no NUP98-HOX fusion genes were detected in the samples from Xinjiang. PMID- 15748442 TI - [STI571 used in treating acute myeloid leukemia with Philadelphia chromosome]. AB - The objective was to use STI571, a kind of tyrosine kinase inhibitor, to treat acute myeloid leukemia (AML) with Philadelphia chromosome. 2 AML cases with Philadelphia chromosome, were collected to observe effect of STI571. One of them was given STI571 after routine chemotherapy failed to respond to treatment, other case received routine chemotherapy only. Both of them underwent HLA matched/mismatched sibling hematopoietic stem cell transplantation (HSCT). The results showed that cytogenetic and hematologic CR was acquired in case 1 with STI571 while another one gained hematologic CR by accepting routine chemotherapy. Recovery of hemopoiesis was found at 18 and 11 days after HSCT respectively without serious graft-versus-host-disease. The case 1 has been surviving in disease-free state for 5 months since HSCT. The case 2 died from interstitial pneumonia at 8 months after HSCT. In conclusion, STI571 is one of choice for the treatment of Philadelphia chromosome positive AML. PMID- 15748443 TI - [Expression of bFGF and VEGF in acute leukemia and its effects on HL-60 cell growth]. AB - The study was to investigate the expression levels of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in the serum of patients with acute leukemia and supernatants of leukemia cell lines as well as effects of VEGF-specific antisense oligodeoxynucleotides (ASODN) on the growth of HL-60 cells. Meanwhile the methods to evaluate the VEGF level in the serum of patients with acute leukemia were explored. The levels of bFGF and VEGF in the serum from 32 patients with acute leukemia and 10 healthy subjects and in the supernatants of 5 various human leukemia cell lines were quantified by means of the enzyme-linked immunosorbent assay (ELISA) and were compared. VEGF levels were evaluated not only without standardization but also after standardized by platelet and finally expressed as VEGF/PLT (pg/10(6)). After with different concentrations of VEGF ASODN, HL-60 cell viability was examined with MTT assay and VEGF levels in supernatants were measured with ELISA, respectively. The results showed that bFGF was detected (3 pg/ml) in 14 out of 32 serum samples from patients with acute leukemia, and the positive (37.5%) was significantly higher than that in healthy controls (10%) (P < 0.01). 3 out of 5 supernanant samples obtained from leukemia cell lines demonstrated positive for bFGF as well. There is no difference of the serum VEGF levels between leukemia patients and healthy controls, but the serum VEGF levels in the serum from leukemia patients were significantly higher than those in healthy controls (P < 0.05) after standardization. 4 out of 5 leukemia cell (U937 excluded) were found to express VEGF in the supernanant. After exposure of HL-60 cells to VEGF ASODN at a concentration of 0.5, 1 and 5 micromol/L for 24 hours, the cell viability gradually dropped down to lower levels (P < 0.05 vs controls). After treatment of HL-60 cells with VEGF ASODN at a concentration of 1, 5 and 20 micromol/L for 24 hours, the VEGF levels in supernatants of target cells decreased (P < 0.05 vs controls). The patients with acute leukemia represented the higher levels of serum bFGF and VEGF than controls. Most of leukemia cell lines expressed bFGF and VEGF at different levels. It is concluded that bFGF and VEGF both have effects on regulations of angiogenesis in acute leukemia, but VEGF plays a pivotal role. VEGF-specific ASODN may have a role in them VEGF expression downregulated. Different results may be obtained in the evaluation of VEGF levels in the serum of patients with acute leukemia if different calculation methods are used. The methods reported can measure leukemia associated VEGF more accurately. PMID- 15748444 TI - [Effect of tetrandrine combined with Droloxifen on the expression of bcr/abl of K562 at both mRNA and protein levels]. AB - To observe the effect of Tetrandrine (tet) combined with Droloxifen (DRL) on the expression of bcr/abl mRNA and P(210) BCR/ABL protein of K562 cell line, after K562 cells were cultured in the medium containing Tet (1 micromol/L), DRL (5 micromol/L) separately or in their combination for some time, the changes of bcr/abl mRNA and protein expression were detected by RT-PCR and Western blot respectively. The results showed that the application of single drug of Tet or DRL had no effect on bcr/abl mRNA and BCR/ABL protein expression in K562 cell line. However, Tet in combination with DRL began to downregulate bcr/abl mRNA and P(210) BCR/ABL expression of K562 cells at 48 h and 72 h, respectively. It is concluded that tetrandrine in combination with Droloxifen can downregulate the expression of bcr/abl mRNA and P(210) BCR/ABL protein and the combination may be involved in the mechanism underlying the reverse effects on multidrug resistance in leukemia. PMID- 15748445 TI - [Protein kinase C inhibitor Go6976 sensitizes arsenic trioxide-induced cell apoptosis in chronic myeloid leukemic cells]. AB - To investigate the As(2)O(3)-chemosensitization of Go6976 in K562 cells by its abrogation of As(2)O(3)-induced G(2)/M cell cycle arrest, K562 cells were treated with As(2)O(3) (5 micromol/L) and Go6976 with various concentrations, the distributions of cell cycles were detected by flow cytometry, the cell viability was observed by trypan blue exclusion test and cell proliferation was tested by MTT assay. The results indicated that having treated by As(2)O(3) for 24 h and 48 h, the proportion of K562 cells in G(2)/M phase were (38.02 +/- 7.70)% and (32.58 +/- 7.43)% respectively, and no obvious cell apoptosis appeared. 50 nmol/L Go6976 combined with As(2)O(3) decrease the proportion of cells in G(2)/M phase to (23.24 +/- 2.93)% and (16.18 +/- 1.60)% respectively and increase the proportion of cells in subG(1) phase to (11.82 +/- 2.31)% and (27.80 +/- 2.89)% respectively. Go6976 abrogated G(2)/M cell cycle arrest induced by As(2)O(3) and increased cell apoptosis in a concentration- and time-dependent manner. Additionally, comparing to the control group, Go6976 combined with As(2)O(3) decreased the cell viability and depressed the cell proliferation, but Go6976 alone showed no same effect on them. In conclusion, the effects of AS(2)O(3) chemosensitization of Go6976 on K562 cells is associated with its abrogation of As(2)O(3)-induced G(2)/M cell cycle arrest. PMID- 15748447 TI - [Study on changes of blood coagulation factors in rats with hemorrhagic shock]. AB - The study was aimed to observe the changes of blood coagulation factors in the SD rats suffered from hemorrhagic shock, and to investigate the mechanism of coagulation cascade reaction in the course of shock. The model of hemorrhagic shock was established. 40 SD rats were randomized into eight groups: pre-shock, and 1, 2, 4, 6, 8, 12 and 24 hours after shock, and the levels of plasma FVIII, vWF, TF, D-dimer, FIB, APTT and PT were detected respectively. The result showed that APTT and PT were gradually prolonged, which were significant within 4-6 hour after shock (P < 0.05). APTT and PT were 59.7 seconds and 30.2 seconds respectively. The level of plasma D-dimer markedly increased, and peaked at 8 hour after shock. The level of fibrinogen, TF, vWF and FVIIIa increased in the initial stage of shock. With the development of shock, fibrinogen markedly reduced from 2nd hour (P < 0.05) and dropped to the minimum at 7 hours after shock. Plasma TF, vWF, FVIII significantly decreased after 6 hours and 8 hours (P < 0.001). The ratios of the consumed coagulation factors: FVIII of (86.1 +/- 1.8)%, fibrinogen of (89.6 +/- 0.6)%, vWF (55 +/- 1.4)%, TF (62 +/- 2.5)%. Thus, coagulation factor I (fibrinogen) and FVIII were preferentially consumed. The extrinsic coagulation pathway was dominantly activated, whereas the intrinsic coagulation pathway played a less important role. Fibrinogen and D-dimer might be valuable for the prognosis of patients suffered from shock. It is concluded that hemorrhagic shock trigger the coagulation cascade reaction, and the coagulation factors are greatly consumed. Unbalance of coagulation system plays an important role in the progress of shock. PMID- 15748446 TI - [Study on the high expression of brain-derived neurotrophic factor in multiple myeloma patients and its possible mechanism]. AB - In order to investigate the expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in multiple myeloma patients and the in vitro and in vivo proangiogenic effects of BDNF, the plasma concentrations of BDNF and VEGF in MM patients and control group were determined by ELISA, the effect of BDNF on the in vitro proliferation of human umbilical vein endothelial cells (HUVEC) was examined by MTT assay; the effects of BDNF on HUVEC migration and tube formation were studied by modified Boyden chamber assay and tube formation assay, respectively. Matrigel plug assay and chorioallantoic membrane assay were used to evaluate the effect of BDNF on angiogenesis in vivo. The results demonstrated that the concentration of BDNF was (4.22 +/- 0.64) ng/ml and (2.03 +/- 0.38) ng/ml in MM group and control group, respectively, (P = 0.01). There was also a significant difference between VEGF levels of two groups [(79.35 +/- 13.25) pg/ml vs (34.41 +/- 1.78) pg/ml, P = 0.006]. The levels of BDNF and VEGF correlated significantly (r = 0.430, P = 0.025). BDNF stimulated the migration and tube formation in vitro significantly, although it had no effect on the proliferation of HUVEC. BDNF also stimulated angiogenesis both in matrigel plug of mouse model and in chick chorioallantoic membrane. It is concluded that the concentrations of BDNF and VEGF in MM patients' peripheral blood are at high level; BDNF can stimulate the angiogenesis markedly in vitro and in vivo. Therefore, BDNF may act as an important regulator in angiogenesis of MM. PMID- 15748448 TI - [Significant decrease of recent thymic output function in patients with severe benzene intoxication]. AB - The aim of the present study was to investigate the level of T-cell receptor excision DNA circles (TREC) in peripheral blood mononuclear cells (PBMNC) of patients with severe benzene poison, thereby to evaluate the content of naive T cells and the recent thymic output function. Quantitative detection of TREC in DNA of PBMNCs from 16 normal individuals and 8 cases with severe benzene poison was preformed by real-time PCR using TaqMan technique. The results showed that TREC level was 6.69 +/- 4.79/1 000 PBMNCs in normal individuals, however, significant decrease was shown in patients with severe benzene poison (1.03 +/- 0.44/1 000 PBMNCs, P < 0.01). The TREC level was persistently low in period of benzene poisoning, even if peripheral blood cell counts were at normal levels. In conclusion, the recent thymic output function is remarkably decreased in patients with severe benzene intoxication, that may obviously damage the T cell immune function. PMID- 15748449 TI - [Effect of phytohemagglutinin on proliferation and cytotoxicity of cytokine induced killer cells]. AB - The purpose was to investigate the effect of phytohemagglutinin (PHA) on proliferation and cytotoxicity of cytokine-induced killer (CIK). Peripheral blood mononuclear cells (PBMNCs) from healthy donors were divided into two groups. Cells were resuspended and maintained in complete medium containing of 10% autologous plasma. CIK cells were cultured by traditional method in group one. The other group cells were added PHA to stimulate PBMNCs for 24 hours, then cultured like incubating CIK cells. Their cytotoxicity to different target cells was evaluated by (51)Cr release assay. The results showed that the proliferation multiples of CIK and PHA-CIK cells were both high, however, the latter was much higher than CIK with significance (P < 0.05). Cells in each group cells showed high cytotoxicity. At the same high effector/target ratio PHA-CIK cells cytotoxicity was stronger than CIK cells when targets were K562 cells or acute leukemia cells (P < 0.05). In conclusion, PHA-CIK cells exhibit stronger proliferation and cytotoxicity than CIK cells, and the result provides an experimental basis for biotherapy. PMID- 15748450 TI - [Effects of blood serum from rats with combined radiation-thermal injury on the bone marrow hematopoietic progenitor cells growth]. AB - To observe the effects of blood serum from rats with radiation injury, thermal injury and combined radiation-thermal lesions on growth of hematopoietic progenitor cells and the change of their serum cytokine levels, total body irradiation of rats was performed with 12 Gy gamma ray from a (60)Co source, and 30% total body surface area III degree thermal lesion on the back was inflicted with a 5 kW bromotungsten lamp. The blood serum from these animals was collected at 3, 12, 24, 48, 72 and 96 hours after injury. Then the blood serum was added to the culture medium of erythrocyte progenitor cells (CFU-E, BFU-E) or granulocyte macrophage progenitor cells (CFU-GM) at final concentration of 10 microg/ml. The results showed that the colony number of CFU-E, BFU-E and CFU-GM formed after addition of the blood serum from rats with thermal or combined radiation-thermal injury was significantly higher than that from normal rats at 3, 12, 24, 48, 72 and 96 hours after injury and reached its peak value at 24 hours after injury (342.8, 261.6 and 228.4% respectively from burned rats, 252.4, 205.1 and 174.2% respectively from rats with combined radiation-thermal injury as compared with that of normal rats). However, a few CFU-E, BFU-E or CFU-GM formation was found after addition of the blood serum from irradiated rats. At the same time, the level of TNF alpha and IL-6 in serum of burn group and combined radiation-thermal injury group was markedly higher than that of normal group, even more higher than that of irradiation injury group (P < 0.01). It is concluded that the blood serum from rats with thermal lesion or combined radiation-thermal injury improves the growth of erythrocyte and granulocyte progenitor cells. On the contrary, the blood serum from the irradiated rats shows the inhibiting effects, definitely related to their serum cytokines changes. PMID- 15748451 TI - [MAPK signal pathway plays a role in proliferation of Jurkat cells induced by ouabain]. AB - The object was to study the effect of ouabain on Jurkat cells and its possible mechanism. The effect of ouabain of low concentration on Jurkat cells was confirmed by MTT, while c-myc gene transcription was measured by RT-PCR, and the phosphorylation of MAPK (ERK1/2) as well as the expression of c-myc gene was tested by Western blot respectively. The results showed that ouabain at low concentration could induce the proliferation of Jurkat in a time-and dose dependent manner. At the same time, the phosphorylation of MAPK (ERK1/2) and the expression of c-myc gene was enhanced. In conclusion, ouabain stimulates the intracellular MAPK signal pathway by acting on the Na, K-ATPase, and thus induce the proliferation of Jurkat cells, in which the regulation of c-myc gene expression may be involved. PMID- 15748452 TI - [Determination of human RHD gene rhesus box and its significance]. AB - The aim was to determine RHD zygosity, further to investigate genetic structure of RHD gene, and to predict hemolytic disease of newborn (HDN). The upstream box, downstream box, and hybrid box of RHD gene were determined by PCR-SSP with 4 primers under the same conditions. The results showed that only hybrid box could be determined in RHD(-)/RHD(-) homozygosity. All the upstream box, downstream box, and hybrid box could be determined in RHD(+)/RHD(-) heterozygosity, while upstream box and downstream box except hybrid box could be determined in RHD(+)/RHD(+) homozygosity. Out of 50 cases of RhD(+), 5 cases (10%) were RHD(+)/RHD(-) heterozygosity, and the others (90%) were RHD(+)/RHD(+) homozygosity. 54 cases (55.1%), 36 cases (36.7%) and 8 cases (8.2%) were RHD( )/RHD(-) homozygosity, RHD(+)/RHD(-) heterozygosity, and RHD(+)/RHD(+) homozygosity respectively in 98 unrelated cases of RhD(-) Chinese Hans. 2 cases of weak D were proved to be RHD(+)/RHD(-) heterozygosity. Out of 16 D(el) types, the upstream box, downstream box, and hybrid box could be determined in 10 cases (37.5%) and the upstream box and downstream box except hybrid box could be determined in 6 cases. Results detecting of RHD 10 exons in above samples proved the correctness of the method. It is concluded that the method is suitable for clinical application with its simplicity and veracity. There are many noneffective RHD genes (44.9%) in Chinese Hans with RhD(-) phenotype. PMID- 15748453 TI - [Molecular genetic analysis for the A3 alleles]. AB - To study four A(3) subgroup samples identified by serologic tests, among which two belong to a family, three were A(3) subgroup, one was A(3)B subgroup. All four samples were genotyped by PCR-SSP method, and the nucleotide sequences of Exon 6, Exon 7 and part introns at the ABO locus for these samples were detected by ABI Prism 3100 DNA sequencer. Comparison with the consensus of A101 was performed. The results showed that haplotypes of two A(3) subgroups were common A102 allele and O1-2 allele, and haplotypes of one A(3) subgroup were common A102 allele and rare O(1v)-4 allele. Unexpectedly, a synonymous substitution 838C-->T had been found in A allele of the A(3)B subgroup sample, which predict a Leu280Phe alteration. The results suggested that molecular genetic background of the A(3) phenotypes is polymorphic. Possibly, the missense mutation 838C-->T is the molecular genetic basis of A(3)B subgroup that lead to low activity of the glycosyltransferases. PMID- 15748454 TI - [Whole exon 5 and intron 5 replaced by RHD/CE in partial D phenotype DVa (Hus)]. AB - The study was purposed to analyze DNA and allele structure of the partial D phenotypes D(Va) and D(VI) of the Rhesus blood group in Chinese. Through polymerase chain reaction (PCR) and direct genomic DNA sequencing, the RHD gene was detected in three weak D individuals identified serologically. The results showed that among the three weak D individuals, one was identified as partial D phenotype D(Va) (Hus) type and genotyped DccEe; another two were testified as D(VI) III type and genotyped DCcee. Moreover, the breakpoints of the replaced region by RHCE in D(Va) (Hus) were 5' end of the exon 5 and 3' end of the intron 5, and there were 7 novel polymorphisms in intron 5: 23-25(GCA)2, 98G>A, 168 169insG, 205-206insT, 494-495insA, 1256-1257insC, 1347G>T. In conclusion the whole exon 5 and intron 5 are replaced by RHCE in D(Va) (Hus) detected in Chinese. PMID- 15748455 TI - Processing and cryopreservation for 1963 units of human umbilical cord blood. AB - The study was aimed to establish a standard procedure for human umbilical cord blood bank. The hematopoietic nucleated cells in cord blood were processed by using sedimentation and centrifugation method. After finishing CD34(+) cell counting, hematopoietic progenitor cell assay, microbial culture, infectious disease test and HLA typing, cord blood units were stored in the liquid nitrogen for further application. The results showed that nucleated cells of cord blood were (10.94 +/- 2.74) x 10(8) per unit; recovery rate of nucleated cells was (79.82 +/- 17.76)%. CD34(+) cells in cord blood were counted as (51.62 +/- 30.53) x 10(5) per unit. Eight units of cord blood were thawed after two years of cryopreservation, the recovery rate of nucleated cells, CD34(+) cells and CFU-GM were (91.4 +/- 6.0)%, (84.6 +/- 20.0)% and (85.8 +/- 14.9)% respectively. It is suggested that the methods and procedure reported for processing and cryopreservation of hematopoietic stem/progenitor cells in the human umbilical cord blood is effective. PMID- 15748456 TI - [G6PD Gene Mutations in Shui people in Sandu of Guizhou]. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. To date, about 126 mutations in the G6PD gene have been detected, among which 17 mutations were found in Chinese. The most common mutations are: 1376 G-->T and 1388 G-->A, both in exon 12; 95 A-->G in exon 2, which amounted to more than 50% of mutations representing various regions and ethnic groups in China. A large-scale screening and genotypic analysis was held in Shui people in Sandu of Guizhou. To investigate the incidence and the molecular basis of G6PD deficiency of Guizhou Shui people, NBT qualitative and G6PD/6PGD quantitative methods were used to detect G6PD deficiency in 1,090 Shui people from the general people belonging to Sandu of Guizhou. By means of mis-matched primers amplified the G6PD gene, the products were 234 bp, 280 bp and 345 bp in length, then restriction enzyme analysis was used to detect the most common Chinese G6PD mutations, 1376 G-->T, 1388 G-->A and 95 A-->G. The results showed that out of the 1,090 samples, 98 G6PD deficiency samples were found. The incidence of G6PD deficiency was 8.99%. 24 cases of 1376 G-->T, 12 cases of 1388 G-->A, 9 cases of 95 A-->G were detected. A sample with 1376 G-->T and 95 A-->G mutation was found in a girl. It was reported for the first time. IN CONCLUSION: 1376 G-->T, 1388 G- >A, 95 A-->G mutations are the common G6PD mutations in Shui people in Sandu of Guizhou. The results indicates that different national minorities of Chinese may originated from a common ancestor. PMID- 15748457 TI - [Clinical efficacy of recombinant human interleukin 11 on thrombocytopenia in pre aplastic anemia]. AB - To evaluate the clinical efficacy of recombinant human Interleukin 11 in the treatment of pre-aplastic anemia, six patients with pre-aplastic anemia were injected with rhIL-11 of 6 million units once a day during 7-14 days. Blood platelet counts were taken on day 8, 15, 30 and 60 after the treatment, and bone marrow examination was performed on day 15 as compared with those before treatment. The results showed that platelet counts in 3 out of 6 patients increased remarkably (50%), one of the six increased moderately (16.7%), another case of the six increased slightly (16.7%), platelet in one out of six did not significantly increase (16.7%), the total efficacy rate is 83.3%, the amount of megakaryocyte in bone marrow of all six patients increased, the side effect of the rhIL-11 treatment was light. In conclusion, the efficacy of recombinant human Interleukin-11 in the treatment of thrombocytopenia patients with pre-aplastic anemia is satisfactory. As the number of the cases is too small to conclude, further exploration needs accumulation of more applications. PMID- 15748458 TI - [Quantification of human ermap by using real-time FQ-PCR]. AB - To develop a real-time FQ-PCR method for quantifying human ermap, a set of primers and a fluorescent probe were designed by primer express 2.0. pBluescriptSK(+) plasmid contained ermap cDNA was transcribed to generate calibration standards for quantification. A real time FQ-PCR method was established. The results showed that when the concentrations of DNA to be amplified were ranged from 1.725 x 10(7) to 1.725 x 10(10) cps/ml, there was a good correlation between template concentration and cycle threshold, and the correlation coefficient reached to -0.999376. In conclusion, real time FQ-PCR which is specific, sensitive and accurate can be used to further research on human ermap. PMID- 15748459 TI - [Application of mesenchymal stem cell in immunotherapy--review]. AB - There has been an increasing interest in recent years on mesenchymal stem cell (MSC). It is well known that MSCs are capable of self-renewal and differentiating into many cell lineages. MSC can be expended to a large quantity that is required for clinical transplantation. Recent studies show that MSC have potential application in immune diseases due to their unique immunologic characteristics, such as low immunogenicity and immunoregulatory function. But their immunoregulatory mechanism is not yet clear. This review discusses the advances in researches on the mechanism of MSCs' immunoregulatory function and potential clinical application in immune disease and organ transplantation. PMID- 15748460 TI - [AGM region and hematopoiesis during ontogeny--review]. AB - During mammalian ontogeny, hematopoietic activity can be found in distinct anatomical sites, which con-tribute to primitive or definite hematopoiesis. The origin of the hematopoietic stem cell (HSC) has been a controversial issue in the field of hematopoiesis. It has long been believed that the origin derives from the extra-embryonic yolk sac. However, there is now considerable evidence that the first adult repopulating HSC is autonomously generated from a distinct region within the embryonic mesoderm, the aorta-gonad-mesonephros (AGM) region. This review describes the origin and precise location of HSC in the embryo and in AGM region, the hematopoietic microenvironment and the hematopoietic regulatory mechanisms in AGM region. PMID- 15748463 TI - Predictors of local recurrence after breast-conservation therapy. AB - Breast-conserving therapy (BCT) is a proven local treatment option for select patients with early-stage breast cancer. This paper reviews pathologic, clinical, and treatment-related features that have been identified as known or potential predictors for ipsilateral breast tumor recurrence in patients treated with BCT. Pathologic risk factors such as the final pathologic margin status of the excised specimen after BCT, the extent of margin involvement, the interaction of margin status with other adverse features, the role of biomarkers, and the presence of an extensive intraductal component or lobular carcinoma in situ all impact the likelihood of ipsilateral breast tumor recurrence. Predictors of positive repeat excision findings after conservative surgery include young age, presence of an extensive intraductal component, and close or positive margins in prior excision. Finally, treatment-related factors predicting ipsilateral breast tumor recurrence include extent of breast radiation therapy, use of a boost to the lumpectomy cavity, use of tamoxifen or chemotherapeutic agents, and timing of systemic therapy with irradiation. The ability to predict for an increased risk of ipsilateral breast tumor recurrence enhances the ability to select optimal local treatment strategies for women considering BCT. PMID- 15748464 TI - Feasibility of quantifying the effects of epoetin alfa therapy on cognitive function in women with breast cancer undergoing adjuvant or neoadjuvant chemotherapy. AB - Impaired cognition, fatigue, and diminished quality of life (QOL) are commonly associated with breast cancer chemotherapy. This randomized, double-blind, placebo-controlled pilot trial assessed the feasibility of quantifying the effects of epoetin alfa on cognitive function and mood, and evaluated its effects on fatigue and QOL in patients with breast cancer treated with anthracycline based adjuvant or neoadjuvant chemotherapy. Patients were randomized to receive epoetin alfa 40,000 U subcutaneously once weekly or placebo at the beginning of 4 cycles of chemotherapy administered over 12 weeks. Cognitive function was assessed by Executive Interview (EXIT25) and Clock Drawing Tasks; mood by Profile of Mood States; anemia-related symptoms, including fatigue, by the Functional Assessment of Cancer Therapy-Anemia (FACT-An) subscale; and QOL by Linear Analog Scale Assessment. Ninety-four patients were evaluable for efficacy and safety. Mean change in EXIT25 scores from baseline to cycle 4 in the epoetin alfa group was 1.3 +/- 3.3; the mean change was 0.3 +/- 2.4 in the placebo group (a negative change indicates improved executive function). There was no difference between groups in mean change in EXIT25 score from baseline to 6-month follow-up assessment. Mean hemoglobin levels were higher in the epoetin alfa group compared with the placebo group after 4 cycles of chemotherapy. Epoetin alfa recipients had less of a decrease in FACT-An subscale scores from baseline to cycle 4 and improvement in FACT-An subscale scores at 6-month follow-up assessment compared with placebo. Epoetin alfa therapy was well tolerated. These data suggest that epoetin alfa may have attenuated the cognitive impairment and fatigue that occurred during adjuvant breast cancer chemotherapy. PMID- 15748466 TI - Management of excessive tearing as a side effect of docetaxel. AB - Docetaxel is a highly effective chemotherapeutic agent for metastatic or locally advanced breast cancer as well as other malignancies. One of the side effects of weekly administration of docetaxel is epiphora (excessive tearing) caused by canalicular stenosis and nasolacrimal duct obstruction. This article summarizes experience with epiphora and canalicular stenosis as a side effect of docetaxel and provides specific recommendations for management of this side effect. PMID- 15748465 TI - Weekly docetaxel/carboplatin as primary systemic therapy for HER2-negative locally advanced breast cancer. AB - To evaluate the effectiveness and safety of weekly docetaxel/carboplatin as primary systemic therapy (PST) for locally advanced breast cancer, we conducted a phase II study. Forty-four patients with HER2-negative locally advanced or inflammatory breast cancer (IBC) received docetaxel 35 mg/m(2) and carboplatin to an area under the curve of 2 mg/mL/min for 3 of 4 weeks over 16 weeks. After completion of PST, patients had breast surgery and then received 4 cycles of adjuvant cyclophosphamide/doxorubicin, standard radiation therapy, and, for hormone receptorpositive tumors, tamoxifen. The mean tumor size was 9.3 cm (range, 5-24 cm). Thirty-seven patients (85%) had palpable lymph nodes; 13 patients (30%) had matted or fixed nodes (N2). Eight patients had IBC. There were 11 clinical complete responses (25%) and 29 clinical partial responses (66%), resulting in 40 objective responses (91% [95% CI, 78%-96%]). Invasive disease disappeared (pathologic complete response) from the breast and axilla in 6 patients (14% [95% CI, 5%-27%]) and from the axilla in 17 patients (39% [95% CI, 24%-55%]). The only significant adverse hematologic event was grade 3 neutropenia in 4 patients (9%). The most common adverse nonhematologic events were fatigue (84% of patients) and alopecia (84%), which were usually grade 1/2. Weekly docetaxel/carboplatin appears to be active and feasible as PST in patients with large breast tumors. PMID- 15748468 TI - [Analysis of surface electromyography on repetitive lifting task-induced fatigue of back muscles]. AB - OBJECTIVE: Using surface electromyography (SEMG) technique to evaluate repetitive lifting task-induced fatigue of back muscles. METHODS: Thirteen volunteers lifted and lowered an 8 kg weight from floor to waist level for 100 times. Fatigue in the erector spinae muscles was quantified by comparing the frequency content of the EMG signal during static contractions performed before, and immediately after the 100 lifts. RESULTS: EMG average amplitude rose gradually during 100 lifts, the difference was significant at T10 right (P < 0.05) and L3 left (P < 0.01), the difference was not significant at T10 left and L3 right (P > 0.05). The median frequency intercept at T10 right, T10 left, L3 right, L3 left erector spinae muscles decreased by 2.0% (P > 0.05) 10.9% and 29.9% (P < 0.05), 27.9% (P < 0.01), respectively. The mean power frequency intercept decreased by 9% at L3 left erector spinae muscle (P < 0.05), the decrease was not statistically significant at other sites (P > 0.05). CONCLUSION: Repetitive lifting may induce measurable fatigue in the erector spinae muscles. Erector spinae muscle at L3 is more easily fatigued than at T10. Using the median frequency intercept to assess muscle fatigue is more sensitive than using mean power frequency intercept. PMID- 15748469 TI - [Estimation of intramuscular load of the upper limb in static postures and repetitive work by surface electromyography]. AB - OBJECTIVE: To evaluate the intramuscular loads of the upper limb during static postures and repetitive work by surface electromyography. METHODS: Twenty-six male college student volunteers were recruited for the experiment. The surface electromyography (SEMG) singal were recorded from the brachioradialis, biceps brachii, deltoid and trapezius of right arm during static postures including forward elevating, abducting, extending and a repetitive performance at different height of the bench, and root mean square (RMS) values were educed from the singal. RESULTS: The SEMG amplitudes from forward elevating and abducting were in direct proportion to the angle of the elevating and abducting (r > 0.9, P < 0.01). The maximal voluntary electrical activation (MVE) of the deltoid were 6.4%, 10.1%, 12.6%, 16.2% and 20.8% while the arm elevated forward at an angle of 0 degrees , 45 degrees , 90 degrees , 135 degrees and 180 degrees respectively. The repetitive work showed that the height of the bench and the duration had more effects on deltoid and trapezius than the other muscles. The MVE% of the deltoid were 13.0%, 14.4% and 15.6% while the bench was 74, 79 and 84 cm in height respectively (P < 0.01). CONCLUSION: SEMG which is suitable for determining and reflecting the muscle strain during static postures and repetitive work may be a reasonable indicator for the assessment of manual workload and the ergonomic design. PMID- 15748470 TI - [Influence of oral sodium selenite and vitamin E on thyroid hormones in patients with cardiovascular disease at altitude]. AB - OBJECTIVE: To investigate the effect of oral sodium selenite and vitamin E on thyroid hormones in patients with cardiovascular disease at altitude. METHODS: Ninety patients with cardiovascular disease were divided into A group (n = 42, sodium selenite + VE), B group (n = 28, sodium selenite only) and control group (n = 20). Serum selenium (Se), plasma glutathione peroxidase (GSH-Px), plasma malondialdehyde (MDA) and serum T(3) and T(4) were determined before and after 6 month treatment. RESULTS: Serum Se in A and B group after 6 month treatment were higher than before [(0.71 +/- 0.22) micromol/L vs (0.31 +/- 0.17) micromol/L, (0.68 +/- 0.18) micromol/L vs (0.33 +/- 0.14) micromol/L respectively, P < 0.01], and so were plasma GSH-Px levels [(87.12 +/- 13.61) U/L vs (58.43 +/- 18.93) U/L, (84.79 +/- 12.13) U/L vs (57.12 +/- 17.36) U/L respectively] while plasma MDA were lower than before [(4.86 +/- 1.18) nmol/ml vs (8.66 +/- 0.96) nmol/ml, (4.18 +/- 1.23) nmol/ml vs (8.71 +/- 0.87) nmol/ml respectively, P < 0.01]. Serum T(3) and T(4) levels in A and B group were also obviously decreased (P < 0.01). The levels of plasma GSH-Px were positively correlated with those of serum Se (r = 0.781, P < 0.01). The levels of plasma MDA and serum T(3) and T(4) were negatively correlated with those of serum Se (r = -0.385, -0.687, -0.412 respectively, P < 0.05). 31 cases (73.81%) in A group and 20 cases (71.42%) in B group completely recovered to normal; 4 cases (9.52%) in A group and 2 cases (7.43%) in B group partly recovered. The recovered rates were significantly different from that of control (P < 0.05). CONCLUSION: Supplementation of adequate selenium may correct the abnormal function of secretion in thyroid hormones of patients because of lack of selenium at altitude areas. PMID- 15748471 TI - [Relationship between heat stress protein 70 gene polymorphisms and the risk of acute mountain sickness]. AB - OBJECTIVE: To investigate the relationship between heat stress proteins 70 (HSPs70) gene polymorphism and the risk of acute mountain sickness. METHODS: Fifty-six soldiers with acute mountain sickness and 173 soldiers without that were chosen as cases and controls. HSP70-1, HSP70-2 genotypes were analyzed by using the polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) technique. RESULTS: The HSP70-1 polymorphism was similar in two groups. The genotype frequency of HSP70-2 B/B in acute mountain sickness group (23.2%) was significantly higher than that in the control (6.9%, P < 0.05, OR = 4.02). CONCLUSION: There is a significantly increased association of HSP70-2 B/B genotype with the risk of acute mountain sickness. Individuals with HSP70-2 B/B genotype may have weaker adaptive ability than those without this genotype under altitude stress. The results contribute to provide scientific bases for finding these individuals in specified occupational people, ensuring their health and enhancing work efficiency. PMID- 15748472 TI - [Investigation on the association of safety perception and safety behaviors with occupational injuries in steel-workers]. AB - OBJECTIVE: To explore the association of occupational injuries, with their individual safety perception and safety behaviors in steel workers, so as to provide basis for preventing and controlling occupational injuries. METHODS: Case control design was used to compare the difference in safety perception and safety behaviors between the injury group and the control, and also to compare the difference in safety behaviors between different safety perception groups. RESULTS: There were remarkable differences in attitude toward the safety degree of the work (chi(2) = 5.444, P < 0.05), and accidents happening (chi(2) = 4.552, P < 0.05) between case group and control group. There were remarkable difference in safety behaviors including manual operations instead of facilities (chi(2) = 10.015, P < 0.01), cleaning up, examining or adjusting machine during work (chi(2) = 7.351, P < 0.05), attention diversion (chi(2) = 10.937, P < 0.01) and unsafe wearing (chi(2) = 7.521, P < 0.05) between case group and control group. There were also significant differences in many safety behaviors between those who thought the job was safe or unsafe. CONCLUSION: There is some association of occupational injuries with safety perception and safety behaviors. To reduce the occurrence of occupational injury, measures should been focused on strengthening safety management and controlling unsafe behaviors. PMID- 15748473 TI - [Relationship of plasma nitric oxide concentration and the activity of nitric oxide synthase with coronary heart disease]. AB - OBJECTIVE: To study the levels of nitric oxide synthase (NOS) and nitric oxide (NO) in patients with coronary heart disease (CHD), and the factors affecting them. METHODS: Concentrations of NO and the activities of NOS in plasma in 136 patients and 206 controls using the corresponding Kits were measured. The data were analyzed using covariance and multiple linear regression analysis with SAS 8.1. RESULTS: The levels of NO [(217.05 +/- 153.31) micromol/L] and NOS [(14.09 +/- 7.14) U/ml] in patients were significantly higher than that in the healthy controls [(140.69 +/- 90.96) micromol/L, (7.75 +/- 3.79) U/ml, respectively, P < 0.01]. Smoking and drinking were the independent risk factors for NOS, while sex was the independent risk factor for NO (P < 0.01). CONCLUSIONS: The levels of plasma NO and NOS are closely related with coronary heart disease. PMID- 15748474 TI - [Measurement of psychosocial factors in work environment: application of two models of occupational stress]. AB - OBJECTIVE: To test the reliability and validity of the Chinese version of Job Content Questionnaire (JCQ) and Effort-Reward Imbalance Questionnaire (ERI) based on the Job Demand-Control Model and Effort-Reward Imbalance Model respectively. METHODS: 928 health care workers in hospitals were investigated in this study. The self-administered questionnaires were used to assess the psychosocial work environment, including job demand, control, social support, extrinsic effort, reward, and overcommitment. Mean while, the internal consistency reliability, construct validity and criterion validity were evaluated. RESULTS: The Cronbach's alpha coefficients for 6 scales were acceptable (0.56 approximately 0.82); factor analysis showed the close fitness with the theoretical constructs, though the "job demand" and "overcommitment" scales were still questioned; moreover, both demand-control and effort-reward imbalance were independently related to depressive symptom (OR: 1.63 approximately 2.64, P < 0.01), indicating the satisfactory criterion validity. CONCLUSION: Both Job Demand-Control Model and Effort-Reward Imbalance Model, with their relevant questionnaires were reliable and valid methods for measuring the psychosocial work environment and assessing the occupational stress among Chinese working population and further revision is necessary. PMID- 15748475 TI - [One case of ethylene dichloride poisoning with wrong diagnosis as epilepsy]. PMID- 15748476 TI - [Norm study of occupational stress on the professionals in Chengdu and Chongqing area]. AB - OBJECTIVE: To establish the norm of occupational stress on the professionals in Chengdu and Chongqing area. METHODS: According to the "Occupation Sorting Canon in the People's Republic of China", the professionals of the above-mentioned area were sorted out and randomly stratified. The normative data were derived from a sample of 2 064 participants. The modified Occupation Stress Inventory Revised Edition (OSI-R) was used to test the targets' occupational stress level (including stressors, strain and coping resources). RESULTS: (1) In these professionals, the sex ratio was about 1:1, average age (36.4 +/- 9.4) years, average length of service (15.9 +/- 10.0) years, 63.07% people were college educated and over 80% married. (2) Descriptive statistics for OSI-R scale scores for the total normative sample, for gender samples and for occupational group samples were modulated. (3) The gender norm showed that there were heavier role overload and stronger occupational strains in males than those in females (P < 0.01). The score of personal resources in the males was not different from that in females (P > 0.05). (4) In terms of different occupations, the scores of occupational role and personal strain among various groups indicated significant differences (P < 0.01). The score of personal resources did not manifest difference (P > 0.05). (5) The major factors influencing OSI-R were sex, occupation and length of service. CONCLUSION: The norm established can basically represent the occupational stress on professionals in Chengdu and Chongqing area. PMID- 15748478 TI - [The mental health situation and its influencing factors in general hospital nurses]. AB - OBJECTIVE: To study the mental health status and its affecting factors in nurses of general hospitals. METHODS: Four hundred and seventy nurses in general hospitals were investigated with SCL-90, Self-Rating Life Events and coping style questionnaires. RESULTS: The scores of SCL-90 factors, such as anxiety, obsessive compulsion, phobia, somatization, were higher than the norm (P < 0.05). 22% of the nurses had two factors higher than 2, and 5.3% ever had suicidal idea. The total score of SCL-90 and some factorial scores in < 40 years age group were higher than in older age groups (P < 0.01), and in III grade hospital nurses were also higher than in II grade hospital nurses (P < 0.01). The scores of negative life events, negative coping style and work load from abnormal psychology group were higher than normal group, while positive coping score was lower than normal group (P < 0.05, P < 0.01). Stepwise regression analysis showed that, negative life events, coping style, hospital grade had predictive value for total score of SCL-90 and anxiety, depression etc factorial scores (P < 0.01). Age had negative predictive value for anxiety, hostility and phobia (P < 0.05). The total score from work events had predictive value for obsession and hostility, and positive events from work had predictive value for anxiety and the total score of SCL-90 (P < 0.01). The total interpersonal relationship events and negative interpersonal relationship events had predictive value for hostility and interpersonal sensitivity. CONCLUSION: General hospital nurses especially in ages < 40 group had higher incidences of mental health problems. The main influencing factors are negative life events, coping style, work load, interpersonal relationship, age and hospital grade. PMID- 15748477 TI - [The scale and application of the norm of occupational stress on the professionals in Chengdu and Chongqing area]. AB - OBJECTIVE: To establish the scale of the norm of occupational stress on the professionals and put it into practice. METHODS: T scores were linear transformations of raw scores, derived to have a mean of 50 and a standard deviation of 10. The scale standard of the norm was formulated in line with the principle of normal distribution. RESULTS: (1) For the occupational role questionnaire (ORQ) and personal strain questionnaire (PSQ) scales, high scores suggested significant levels of occupational stress and psychological strain, respectively. T scores >/= 70 indicated a strong probability of maladaptive stress, debilitating strain, or both. T scores in 60 approximately 69 suggested mild levels of maladaptive stress and strain, and in 40 approximately 59 were within one standard deviation of the mean and should be interpreted as being within normal range. T scores < 40 indicated a relative absence of occupational stress or psychological strain. For the personal resources questionnaire (PRQ) scales, high scores indicated highly developed coping resources. T scores < 30 indicated a significant lack of coping resources. T scores in 30 approximately 39 suggested mild deficits in coping skills, and in 40 approximately 59 indicated average coping resources, where as higher scores (i.e., >/= 60) indicated increasingly strong coping resources. (2) This study provided raw score to T score conversion tables for each OSI-R scale for the total normative sample as well as for gender, and several occupational groups, including professional engineer, professional health care, economic business, financial business, law, education and news. OSI-R profile forms for total normative samples, gender and occupation were also offered according to the conversion tables. CONCLUSION: The norm of occupational stress can be used as screening tool, organizational/occupational assessment, guide to occupational choice and intervention measures. PMID- 15748479 TI - [Studies on the microwave leakage of the interphone]. AB - OBJECTIVE: To assay and study the microwave leakage of 4 types of interphones. METHODS: The radiation intensities of four types of 199 interphones were determined by the microwave leakage measure instrument of model ML-91 made in China. RESULTS: The average intensities of microwave leakage at a distance of 5 cm from aerial part and other parts of interphones during launching [(1 316.0 +/- 144.3), (971.0 +/- 131.6) microW/cm(2) respectively] were significantly higher than during waiting [(14.4 +/- 5.3), (13.2 +/- 4.9) microW/cm(2) respectively] (P < 0.01). The average intensities of microwave leakage at a distance of 50 cm from different parts were (357.3 +/- 27.8) microW/cm(2). The daily average intensity of microwave leakage to which the head, chest and abdomen exposed was (945.5 +/- 447.1) microW.h/cm(2) in total, that exceeded the hygienic standard of microwave in China (400 microW.h/cm(2)), during the normal communication by interphones. CONCLUSION: The microwave leakage was higher during launching than during waiting, and was the highest at the aerial part of the interphones. The microwave radiation of most interphones was higher than the current national standard. It may lead to potential effects on the owner of interphone, so protection against it should be made. PMID- 15748480 TI - [Protection of zinc on hippocampal cholecystokinin of lead-exposed rat]. AB - OBJECTIVE: To investigate the relationship between the effects of zinc on hippocampal cholecystokinin (CCK) positive neurons and learning and memory ability of lead-exposed rats. METHODS: Thirty-six Wistar rats were divided into control group, lead-exposed group (drunk 6.15 mmol/L of lead solution) and lead zinc group (drunk 6.15 mmol/L of lead + 3.10 mmol/L of ZnSO(4) solution) randomly. Y-maze test was used to study learning and memory ability in rats; Atomic absorption method was used to determine serum and hippocampal lead content; ABC immunohistochemistry and quantitative graphic analysis were used to investigate the changes of CCK positive neurons in different hippocampal subfields in lead-exposed rats. RESULTS: The learning and memory ability in lead exposed rats were significantly lower (P < 0.05) while the serum and hippocampal lead content in lead-exposed rat were significantly higher (P < 0.05) than those in control and lead-zinc group. The number and optical density of CCK positive neurons in CA(1) and CA(3) areas of lead-exposed rats were significantly lower (P < 0.05) than those in control and lead-zinc group. No differences in these indexes between the control and lead-zinc group were found (P > 0.05). CONCLUSION: Lead may damage the learning and memory ability and affect the number of CCK positive neurons in lead-exposed rats. Zinc might play an important role in preventing lead-induced damages. PMID- 15748481 TI - [Effects of selenium on benzo[a] pyrene-induce DNA damage in mouse lung cells]. AB - OBJECTIVE: To explore the effects of selenium on DNA damage induced by benzo[a] pyrene (BaP) in mouse lung cells. METHODS: Sodium selenite was given to Kunming male mice by i.p. and BaP was given by oral gavage. The control group was given solvent only with the same method. DNA damage was detected by single cell gel electrophoresis (or comet assay). RESULTS: The damage degrees in mice treated with 125, 250 and 500 mg/kg of BaP were more severe than that of control (P < 0.01). The rates of comet cells in the BaP-treated groups (43.50%, 84.00%, 95.63%) were significantly higher than that of control (9.75%, P < 0.01), and there was obvious dose-response relationship. 0.75, 1.50 and 3.00 mg/kg of sodium selenite presented antagonistic effects against DNA damage induced by 250 mg/kg of BaP in mouse's lung cells. The antagonistic effect of sodium selenite at the dose of 1.50 mg/kg was better than those of sodium selenite at the doses of 0.75, 3.00 mg/kg. CONCLUSION: BaP at the doses of 125 approximately 500 mg/kg could significantly induce DNA damage of lung cells in mice. 0.75 approximately 3.00 mg/kg of sodium selenite could inhibit DNA damage of lung cells in mice induced by 250 mg/kg of BaP. PMID- 15748483 TI - [Effect of video terminal operation on the health of operators]. PMID- 15748482 TI - [Inhibitory effect of grape procyanidin on the cell apoptosis induced by radiation]. AB - OBJECTIVE: To study the inhibition effect of grape procyanidin (GPC) on the cell apoptosis and injury of proliferation induced by radiation. METHODS: Three indices including apoptosis rate, proliferation rate and expression of bcl-2 and bax protein were examined in the mice pancreas after taking different dose GPC by mouth and radiation by (60)Co-gamma ray once. RESULTS: The cell proliferation and bcl-2 expression in high dose GPC group (3.16% +/- 0.13% and 49.8% respectively), were higher than those in radiation control group (0.64% +/- 0.11%, 29.7%), but the cell apoptosis rate and bax expression (19.8% and 55.0% respectively), were lower than those in radiation control group (35.6%, 85.7%). All the above differences were statistically significant (P < 0.05). CONCLUSION: GPC has certain protective effect against the mice pancreatic cell apoptosis and the abnormal expression of bcl-2 and bax protein induced by (60)Co-gamma. PMID- 15748484 TI - [Relationship between female occupational stress and work ability in oil-field]. PMID- 15748485 TI - [The exposure level of microwave radiation for the workers enclosed in metal separation rooms in the workplace]. PMID- 15748486 TI - [Follow-up study on the patients with chronic radiation sickness by external irradiation]. PMID- 15748487 TI - [Physical activity prevalence study in Shanghai city]. PMID- 15748488 TI - [Investigation on accidents happened during 1966-2003 in one petrochemical enterprise]. PMID- 15748489 TI - [Development of rat model of seasickness adaptation]. PMID- 15748490 TI - [Cloning of the differentially expressed genes from the brain stem of seasickness adaptive rats]. PMID- 15748491 TI - [Clinical analysis on the bone marrow cell picture of benzene-induced acute monocytic leukemia]. PMID- 15748492 TI - [Determination of myocardial enzymes to assess acute CO poisoning-induced myocardial damage]. PMID- 15748493 TI - [Three cases of cholesteatoma otitis media in pilots]. PMID- 15748494 TI - [A case of tetramine poisoning-induced basal ganglion damage]. PMID- 15748495 TI - [Changes of cardiac pathology in patients with cor pulmonale and coal workers' pneumoconiosis]. PMID- 15748496 TI - [Accurate comprehension and enforcement of "Diagnostic Criteria of Occupational Hand-arm Vibration Disease"]. PMID- 15748497 TI - [Method for detecting boron in the animal's urine and bone ashes]. PMID- 15748498 TI - [Effect of military solid-sport-beverage on expression of C3b-receptors on macrophage surface]. PMID- 15748499 TI - [Assessment of extreme low frequency magnetic field from some domestic instruments]. PMID- 15748500 TI - [Review on carpal tunnel syndrome induced by keyboard operation]. PMID- 15748501 TI - [Genetic polymorphisms of NQO1, GSTT1, GSTM1 and susceptibility to chronic benzene poisoning]. AB - OBJECTIVE: To explore the relationship between genetic polymorphism of quinone oxidoreductase 1 (NQO1), glutathione S-transferase theta 1 (GSTT1), glutathiones S-transferase mu 1 (GSTM1) and susceptibility to chronic benzene poisoning (BP). METHODS: The genotypes of NQO1, GSTT1, GSTM1 for 100 patients with benzene poisoning and 90 workers exposed to benzene who were engaged in the same working time and job title as patients with benzene poisoning were detected by PCR-RFLP and multi-PCR. RESULTS: There was a 2.82-fold (95% CI: 1.42 approximately 5.58, P < 0.05) increased risk of BP in the subjects with NQO1 C609T mutation genotype (T/T) compared with those carrying heterozygous (C/T) and wild type (C/C), and there was a 2.94-fold (95% CI: 1.25 approximately 6.90, P < 0.05) increased risk of BP in the subjects with NQO1 C609T T/T genotype compared with those carrying C/C genotype. The subjects with GSTT1 null genotype had a 1.91-fold (95% CI: 1.05 approximately 3.45, P < 0.05) increased risk of BP compared with those with GSTT1 non-null genotype. The interaction of two genes showed that there was a increased risk of BP in subjects with any two genotypes of NQO1 C609T T/T genotype and GSTT1 null genotype and GSTM1 null genotype, compared to the individual with any two genotypes of NQO1 C609T C/C genotype and GSTT1 non-null genotype and GSTM1 non-null genotype. The interaction of three genes showed that there was a 20.41 fold (95% CI: 3.79 approximately 111.11, P < 0.01) increased risk of BP in subjects with NQO1 C609T T/T genotype and GSTT1 null genotype and GSTM1 null genotype compared with those carrying NQO1 C609T C/T genotype and C/C genotype and GSTT1 non-null genotype and GSTM1 non-null genotype. CONCLUSIONS: The interaction of multi-genes may be an important role to BP. The genetic polymorphisms of 3 genes (NQO1, GSTT1 and GSTM1) led to declining of detoxifying ability in benzene metabolism, so the individual with NQO1 C609T T/T genotype, GSTT1 null genotype and GSTM1 null genotype is most susceptive to benzene. The results were consistent with that of the theoretic presumption. It could be suggested as a biomarker to assess the risk of benzene poisoning for individuals. PMID- 15748502 TI - [Effects of ethylene oxide and propylene oxide on human health]. AB - OBJECTIVE: To study the hazards of demulsifier production process. METHODS: The air of production environment was monitored, and physical examinations were conducted on 52 exposure workers and 53 non-exposure workers (control), all the medical checks including clinical, electromyography and laboratory checks. RESULTS: The air concentration of propylene oxide was 4.6 mg/m3, and the concentration of Ethylene oxide was 4.8 mg/m3. There was significant difference between exposure group and control group in terms of the incidence of neurasthenic syndromes, lachrymation, laryngopharyngeal ache, inappetence, tired and quadriplegia (P <0.01). The incidence of Achilles's tendon reflection, knee jerk slacking up, chronic pharyngitis, coryza, acne, hair lose and pneumonia were significantly higher in study group than that in control group (P <0.01). Peripheral nerve damage of impairment incidence rate was 40.4%, significantly higher than that in control group (9.4%). No significant difference was found between groups on IgG, IgA and IgM. And also did the rate of chromosome aberrance (CA), sister chromatid exchange (SCE) and micronuclie cell (MC) of peripheral blood (P >0.05). CONCLUSION: The production environment of demulisier could stimulate respiratory channel and affect peripheral nerve and central nerve, but no aberrance effects were found. PMID- 15748503 TI - [Comparing three methods of estimating concentration of worksite benzene.]. AB - OBJECTIVE: benzene; Air pollution; Risk assessment in the air of workshop and validate the methods. METHODS: Expert-reference, BAYES statistics and expert measurement were separately used to estimate benzene concentration in a slurry applying workshop and modeling workshop of a rubber factory. The estimations were compared with current newly monitored data. RESULTS: The estimated averages of expert-reference, BAYES statistics and expert-measurement were 100.0, 16.9, 33.2 mg/m3 respectively for the slurry-applying workshop and 100, 156, 115 mg/m3 for the modeling workshop. No significant difference was observed between the estimations and validating sample except to that of the expert-reference for the former. CONCLUSION: BAYES statistics and expert-measurement were precise and expert-reference was candidate if no measurements in spite of its subjectivity. PMID- 15748504 TI - [The intervention of nicotinamide on skin melanocyte's cell proliferation after UVA (365 nm) exposed.]. AB - OBJECTIVE: To investigate the interference effect of nicotinamide on UVA-induced cell proliferation in human skin melanocyte. METHODS: To apply the optimum UVA dose expected to cause cell proliferation: 0.2 cm2, nicotinamide was added after the 0.2 cm2 UVA exposure immediately or 48 h later, then the rate of cell proliferation, calcium concentration and the activities of Na+-K+, Ca2+-ATP enzymes of melanocytes were measured respectively. RESULTS: After treatment with 1.000 mg/ml nicotinamide following UVA exposure, the rate of cell proliferation was decreased significantly 24 hours later. Treatment with 0.125 mg/ml nicotinamide 48 hours after UVA exposure also significantly inhibited the cell proliferation; 1.25 mg/ml nicotinamide increased calcium concentration in cells; 0.250 mg/ml nicotinamide increased the activities of Na+-K+, Ca2+-ATP enzymes in melanocytes (P < 0.05). CONCLUSION: Nicotinamide has more obvious effect on inhibiting melanocyte's proliferation if added immediately following UVA exposure. Our discovery indicated that nicotinamide may affect the melanocyte through modulating the calcium concentration. It is possible to consider nicotinamide as an efficient and safe sun screen to provide a certain level of protection for UVA exposed skin. PMID- 15748505 TI - [Effect of cadmium on estrogen receptor from rat uterus in vitro.]. AB - OBJECTIVE: To investigate the effect of cadmium (Cd) on estrogen receptor and to assess its endocrine disrupting action. METHODS: The estrogen receptor rich supernatant was prepared from the ovariectomized Sprague-Dawley rats. The effects of cadmium on estrogen binding were performed using a sing-dose ligand-binding assay. Extract from uterus were treated with various concentrations of cadmium (0, 10(-3), 10(-5) or 10(-7) mol/L) for various pre-incubation time (0, 30, 60, 90 min) by means of orthogonal experimental design with orthogonal layout of L16(4(5)) (the experiment was repeated for 5 times). In addition to the radioinert competitor, each assay included a zero tube and a DES standard curve for quality control purpose. Data for cadmium and the DES standard curve were plotted as percent [3H]-E2 bound versus log (molar concentration), and the IC50 for cadmium was determined. The RBA for cadmium was calculated by dividing the IC50 of DES in terms of the IC50 of cadmium. RESULTS: Cadmium could not block the binding of estradiol to the receptor because hormone binding did not change with increasing cadmium concentration or increasing preincubation time. The results showed that the binding of [3H]-estradiol to uterine cytosols was not significant (P > 0.05). The Bmax (its unit is pmol/mg protein) of various concentrations of cadmium (0, 10(-3), 10(-5) or 10(-7) mol/L) for pre-incubating 0 min is 203.15 +/ 75.16, 203.41 +/- 22.78, 220.82 +/- 45.35, 209.10 +/- 49.66 respectively; The Bmax of them for pre-incubating 30 min is 215.67 +/- 92.97, 139.79 +/- 53.78, 205.27 +/- 23.60, 172.63 +/- 55.09 respectively. The Bmax of them for pre incubating 60 min is 197.11 +/- 50.68, 203.24 +/- 66.33, 183.92 +/- 31.89, 183.33 +/- 32.70, respectively. The Bmax of them for pre-incubating 90 min is 229.69 +/- 76.88, 175.70 +/- 70.28, 164.26 +/- 24.46, 150.78 +/- 65.97 respectively. Mean IC50 for cadmium is 10(-4) - 10(-3) M. If the affinities of DES binding to estrogen receptors was taken to be 100%, the relative binding affinities of cadmium was 10(-6) - 10(-7). The results indicated that cadmium had only a very poor affinity with estrogen receptor. CONCLUSION: In vitro assay cadmium did not have distinct disrupting effect on binding of estradiol to estrogen receptors from rat uterine. PMID- 15748506 TI - [Observation on the protective effect of hyperoxia solution on the acute lung injury caused by phosgene poisoning.]. AB - OBJECTIVE: To study the protective effect of hyperoxia solution on acute lung injury caused by phosgene poisoning by observing the changes of PaO2 and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity in serum and Glutathione (GSH/GSSG) contents in lung tissues. METHODS: The rabbits were divided into normal control group, hyperoxia solution (H0) and balance salt (BS) groups. Group HO and Group BS inhaled phosgene and the former was given intravenously hyperoxia solution (which was replaced by balance salt solution in Group BS). The content of MDA and the activity of SOD in serum were observed at different time points, the amount of GSH and GSSG in lung tissue were also measured. RESULTS: (1) The serum MDA contents increased and PaO2, SOD activity decreased significantly in Group HO and Group BS along with time increasing as compared with control group. The contents of GSH in lung tissue decreased in two groups compared with that in control group, however the contents of GSSG ascended instead. (2) At 3 and 8 h of the experiment, PaO2 of Group HO [(9.91 +/- 0.49), (9.15 +/- 0.46) mm Hg respectively] were significantly higher than those of Group BS [(9.03 +/- 0.76), (8.11 +/- 0.57) mm Hg respectively] (P < 0.01). The contents of MDA of Group HO (3.66 +/- 0.35), (5.31 +/- 0.15) micromol/L respectively] were lower than those of Group BS [(4.32 +/- 0.26), (7.4 +/- 0.33) micromol/L respectively] (P < 0.01). SOD activity in Group HO [(237.37 +/- 29.96), (208.10 +/- 18.80) NU/ml respectively] were higher than those of Group BS [(195.02 +/- 21.44), (144.87 +/- 21.26) NU/ml respectively] (P < 0.05 or P < 0.01). The content of GSSG lung tissue in Group HO (423.67 +/- 38.21) micromol/L were lower than those of Group BS (523.85 +/- 43.14) mol/L (P < 0.01). There were no significant differences in the content of GSH in lung tissues between Group HO and group BS. CONCLUSION: Hyperoxia solution can reduce acute lung injury of rabbits following phosgene poisoning. PMID- 15748508 TI - [The effect of lead exposure in utero on the teeth eruption and enamel development of rat offspring.]. AB - OBJECTIVE: To investigate the effects of lead exposure at different levels in utero on the teeth eruption and enamel development of rat offsprings. METHODS: 27 pregnant SD rats were divided into three groups randomly: high level lead group (HLG), low level lead group (LLG) and control group with nine rats in each group. The three groups from the gestation day to the end of the gestation were given either deionized water in control group or deionized water containing 200 mg/L Pb2+ as lead acetate through drinking method in high level lead experimental group and 50 mg/L Pb2+ as lead acetate through drinking method in low level lead experimental group. The incisors of newborn rats were marked at the level of the gingival papilla on the 26th day after birth. On the 36th day, the incisors of newborn rats were marked again at the same level. Then the rat offsprings were killed and their blood was collected for lead analysis. The mandible incisors of rat offspring were separated and the content of Pb in incisors was determined by using a graphite furnace atomic absorption spectrometric method. The teeth of rat offspring were observed and the distance between two marks were measured by means of stereomicroscope. The ratio of calcium to phosphate of enamel of rat offspring was compared by electron probe microanalyses. RESULTS: The level of blood lead in 200 mg/L, 50 mg/L treated rat offspring groups was higher than that in control group. The tooth lead of 200 mg/L, 50 mg/L treated rat offspring groups [(77.3 +/ 6.3), (27.8 +/- 4.5) microg/g] were higher than the control [(6.6 +/- 0.8) microg/g, P < 0.01]. Compared with the control group, the teeth of lead exposure experimental groups were smaller and severity of attrition was obvious and pulpal perforations were often observed. These appearances was more distinct in rats of high level lead experimental group. The incisors of lead-treated rat offspring erupted [(0.25 +/- 0.08), (0.30 +/- 0.09) mm/d] more slowly than control ones [(0.39 +/- 0.09) mm/d, P < 0.01]. The ratio of calcium to phosphate (Ca/P) decreased with the increase of lead exposure. It was found that Ca/P in lead exposure experimental groups (1.68 +/- 0.54), (1.37 +/- 0.47) was significantly lower than that in the control group (2.14 +/- 0.33). CONCLUSION: Lead exposure in utero affects the normal eruption of teeth and enamel formation and the degree was related with the lead exposure level. PMID- 15748507 TI - [A study on relationship between rat brain tissues hurts of the gas explosion and the expression of protein kinase CamRNA and c-fos gene.]. AB - OBJECTIVES: To explore relationship between rat brain tissues hurts of gas explosion and the expression of Protein Kinase C alpha mRNA. METHODS: Build up rat hurt model of gas explosion. In Situ Hybridization (IDH) technique was used to test Protein Kinase C alpha mRNA. Immunohistochemical Assays (IHA) was used to determine c-fos gene protein. RESULTS: Only a little a mount expression of PKC alpha mRNA and c-fos of the control group was detected. The expression of the cerebral cortex and hippocampus of PKC alpha mRNA 24 h, 48 h and the 48 h increased obviously, and the 48 h reached the peak of expression; (t = 4.12 P < 0.01). The expression of c-fos protein of the cerebral cortex and hippocampus started to increase obviously at 0.5 h and the 4 h reached the peak, then the strength lowered gradually and the expression level came back normal level on fifth day. CONCLUSION: The anoxia of brain tissues due to the gas explosion may promote the expression of PKCamRNA, and PKCamRNA could regulate the expression of the gene of c-fos. Both PKCamRNA and the gene of c-fos are involved in harmful processes to the nerve cells. PMID- 15748509 TI - [Expression of protein p53 in workers occupationally exposed to benzidine and bladder cancer patients.]. AB - OBJECTIVE: To study expression of mutant p53 protein in workers occupationally exposed to benzidine and bladder cancer patients. METHODS: Mutant p53 protein in serum from the workers occupationally exposed to benzidine and bladder cancer patients were determined with Immuno-PCR, while exfoliated urothelial cells in the urine samples were classified with Papanicolau grading. RESULTS: Positive rate of mutant p53 protein increased with the exposed intensity index in workers occupationally exposed to benzidine. The positive rate of mutant p53 protein in bladder cancer patients (83.3%) was significantly higher than that in the group 1 of exposed intensity index. The average scanning integrals of PCR amplified band in the group of bladder cancer patients and group 2 of exposed intensity index were both higher than that in the group 1 significantly. Workers in the groups of different exposed intensity indices were further stratified according to Papanicolau grades. In the group 2 of exposed intensity index, the average scanning integrals of PCR amplified band in the stratum of Papanicolau grade II and III were significantly higher than that in the strata of Papanicolau grade I. And in the group 3 of exposed intensity index, the positive rate of mutant p53 protein in the strata of Papanicolau grade III was higher than that in the strata of Papanicolau grade I significantly. CONCLUSION: The increase of exposed intensity may not only result in the positive rate of mutant p53 protein, but also the quantity of mutant p53 protein in serum within the low range of benzidine exposure. Once the exposed intensity was beyond that spectrum, the positive rate of mutant p53 protein in serum and the average scanning integrals of PCR amplified band were no longer enhanced with the increase of exposed intensity. There was tight correlation between Papanicolau grade of exfoliated urothelial cells and the positive rate or the quantity of mutant p53 protein for the higher benzidine exposure intensity. PMID- 15748511 TI - [The delayed protection of exercise preconditioning from the relative myocardial ischemia-reperfusion injury.]. AB - OBJECTIVE: To explore the delayed protection of exercise preconditioning from the relative myocardial ischemia-reperfusion injury. METHODS: The experiment included the vivo experiment and the vitro experiment, 32 Wistar rats in each experiment were divided into 4 groups randomly: control group (CN), relative ischemia reperfusion group (IR), exercise preconditioning group (EP) and Exercise preconditioning + relative ischemia-reperfusion group (EI). We detected the third loading exercise time, the levels of MDA in serum in vivo experiment and the Cardiac function parameter, the levels of MDA in coronary effluent in vitro experiment. RESULTS: (1) The vivo experiment: The third loading exercise time of EI group [(71.67 +/- 9.00) min] increased significantly compared with that of IR group [(58.67 +/- 4.13) min] (P < 0.05); The levels of MDA in serum of EP group (107.00 +/- 35.99) micromol/L and EI group [(152.23 +/- 29.94) micromol/L] decreased significantly contrasted to IR group (313.20 +/- 43.40 micromol/L) (P < 0.05). (2) The vitro experiment: The PRP (heart rate * left ventricular developed pressure) in reperfusion period of CN group and EP group were stable relatively, while it reached the peak after 30 minutes and almost recovered to the level before ischemia in EI group. The parameter of IR group recovered slightly but was lower significantly than that before ischemia. There was significant difference between the recovery rate of Cardiac function of EI group and that of IR group. The increase of MDA in coronary effluent after Ischemia-reperfusion of EP group (0.34 +/- 0.24 micromol/L) and EI group [(0.41 +/- 0.26) micromol/L] decreased significantly contrasted to that of IR group [(1.27 +/- 0.52) micromol/L] (P < 0.05). CONCLUSION: EP has the obvious delayed protection from the relative myocardial ischemia-reperfusion injury. PMID- 15748510 TI - [The ultrastructure change of heart capillary in athlete's heart.]. AB - OBJECTIVE: To see the change of capillary of heart in Athlete's Heart, so that to discover the mechanism of pathologic change. METHOD: 18 male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (swimming for 75 min every day), and overload group (swimming for 180 min with 5% weight of its body every day). After 5 days per week, 12 weeks, exercise training stopped and heart of rats were observed under Transmission Electron Microscope. RESULTS: In aerobic exercise group, the capillary cavities in heart expand, the walls of capillary become thick; the number of mitochondrion increases; endothelium cells become active in growth. However, after overload exercise, the walls of capillary cockle and protuberances appear. The mitochondrion swell and the cristae become disorder. Most of endosomes expand and their number increases. The karyons become abnormity in shape and uniformity in electronic density, besides the nuclear envelope cockle. The basilar membranes become thick and unclear. CONCLUSION: After exercise training, both physical and pathologic changes in heart capillary are found. In suitable exercises group, the capillaries change physically; the pathologic changes are becoming visible after overload exercise however. PMID- 15748512 TI - [Study on the genotoxicity of exhausts of diesel engine with ethanol-diesel blending fuel]. AB - OBJECTIVE: To study the genotoxicity of components of diesel engine exhausts with ethanol-diesel blending fuel. To provide scientific arguments to find more economical and less polluted fuels. METHODS: Ames test, comet assay and GC-MS technique were used to test the genotoxicity and 16 kinds of PAHs on diesel engine exhausts with different proportions of ethanol (E0, E5, E10, E20). RESULTS: Both Ames test and comet assay were positive. It shows that diesel engine exhausts can lead to mutation and DNA damage, especially in pure diesel oil. But the content of 16 kinds of PAHs and DNA damage level decreased in exhausts of E5. With the increase of ethanol proportion in diesel oil, the content of 16 kinds of PAHs and DNA damage level increased. CONCLUSION: Compared with pure diesel oil and high proportion of ethanol fuel, E5 can reduce the genotoxicity and the brake specific exhausts of PAHs. PMID- 15748513 TI - [The toxic effects of cadmium on pancreas.]. AB - OBJECTIVE: In order to explore the toxic effects of cadmium on functions of endocrine and exocrine of pancreas. METHODS: 96 SD rats were administered with cadmium at different doses (0, 50, 100, 200 mg/L) by drinking water for 30 days, 60 days and 90 days. The contents of cadmium and zinc in the blood and pancreas, also the glucose level in blood and urine, the levels of insulin and the activity of amylase were determined. The gene expression of metallothionein (MT), insulin and pancreatic amylase were also measured. RESULTS: The results showed that the contents of cadmium in the serum and pancreas were higher than that of the control groups (P < 0.05). The contents of zinc in serum were decreased in the groups of 100 and 200 mg/L cadmium at the 90-day. As well as increased zinc in pancreas. The gene expression of insulin was not different compared with those of the control group except the middle-dose group at the 60-day. And the expression of amylase were higher in the groups of 100 and 200 mg/L cadmium at the 60-day and the 90-day. The expression of MT-1 and -2 were higher in the pancreas after cadmium administration. CONCLUSION: It is suggested that cadmium could be accumulated in the pancreas and caused the change of the zinc levels. Then it resulted in the change of the expression of gene and protein, and influence of the functions of both endocrine and exocrine in pancreas. PMID- 15748514 TI - [The study on pathogenesis of latex allergy.]. AB - OBJECTIVE: In order to study the pathogenesis of latex allergy and the significance in the prevention and cure of occupational diseases. METHODS: 651 cases in the out-patient department were tested with skin prick test (SPT), and the specific IgE (sIgE) to latex were detected by means of disk ELISA and Western blot. RESULTS: It was found that the positive rate of latex SPT (37.5%) and sIgE (31.25%) were rather higher in patients in comparison with those of the normal. The positive rate of latex sIgE was much higher in the high-risk group than that of the low-risk group and the normal. The serum of the patients can react with multi-bands in the latex glove extracts. CONCLUSION: The prevalence of latex allergy is rather high, this disease is mediated by IgE. The people in high-risk should be tested by latex allergy in order to take proper occupational and daily protection. PMID- 15748515 TI - [Study on occupational health hazards in building Qingzang railway and effectiveness of preventive measures.]. PMID- 15748517 TI - [Study on thigh-complex-potentials of rats immersed in hypothermic sea water.]. PMID- 15748518 TI - [Review on the methods of occupational exposure assessment.]. PMID- 15748519 TI - [Liver toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin.]. PMID- 15748520 TI - [Review on job burnout.]. PMID- 15748521 TI - [Risk of leukemia related with benzene and chromosomal aberrations.]. PMID- 15748522 TI - [Determination of methanol and ethanol in serum by gas chromatography.]. PMID- 15748523 TI - [The Test kit for biochemical identification and drug sensitivity of bacteria]. PMID- 15748524 TI - [One case of encephalopathic caused by lead poisoning]. PMID- 15748527 TI - [One case of hepatic disease by dimethylformamide poisoning.]. PMID- 15748528 TI - [56 cases of acute acrylonitrile poisoning]. PMID- 15748529 TI - Pathophysiology and clinical meaning of small resistance artery remodeling. PMID- 15748530 TI - The metabolic effects of angiotensin-receptor blockers. PMID- 15748533 TI - The metabolic syndrome in the asian population. AB - With the rapid emergence of obesity in Asia, the metabolic syndrome is becoming a clinically important issue. The studies on migrants from Asian countries to Western countries indicate that Asian populations may be more susceptible to the Western diet and lifestyle than whites in developing the metabolic syndrome. The World Health Organization (WHO) Expert Consultation recently reported that the cut-off point for observed risk in Asian populations may be smaller than that for non-Asian populations, varying from 22 kg/m(2) to 25 kg/m(2). This variant susceptibility may be caused by gene-environmental interactions and low birth weight in Asian populations. PMID- 15748532 TI - Advanced-glycation end products in insulin-resistant states. AB - Insulin resistance is a central component of a number of clinical conditions, including the metabolic syndrome, diabetes, and hypertension. There is emerging evidence that the consequent hyperinsulinemia and visceral adiposity may be directly responsible for the excess cardiovascular morbidity and mortality seen in these conditions. Advanced-glycation end products, a chemically diverse group of compounds found in higher levels in insulin-resistant states, have also been shown to adversely affect endothelial function as well as activate numerous intracellular signaling pathways implicated in the atherosclerotic pathway. In this review, we summarize the factors thought to be important in both the initiation and exacerbation of the insulin-resistant state, and directly examine the potential role of advanced-glycation end products in this process. PMID- 15748531 TI - Metabolic syndrome and endothelial dysfunction. AB - The incidence of metabolic syndrome is rapidly increasing in the United States. Metabolic syndrome is associated with increased cardiovascular morbidity and mortality, and endothelial dysfunction is an early pathogenetic event in the metabolic syndrome. Endothelial dysfunction of either the coronary, the peripheral, or the cerebral vasculature is a predictor of vascular events and appears to be a marker of uncontrolled atherosclerotic risk that adds to the burden of the genetic predisposition to cardiovascular disease. Clinically and experimentally, endothelial dysfunction can be restored by several agents, including blockers/inhibitors of the renin-angiotensin-aldosterone system, as well as statins. Nevertheless, it would be premature, and most likely inappropriate, to use improvement of endothelial function as a surrogate end point to predict reduction in cardiovascular morbidity and mortality. However, a clear understanding of the mechanisms of endothelial dysfunction in the metabolic syndrome may allow the development of preventive and early therapeutic measures targeting cardiovascular disease. PMID- 15748534 TI - Metabolic syndrome therapy: prevention of vascular injury by antidiabetic agents. AB - More than 65 million Americans are currently obese. Type 2 diabetes mellitus, frequently seen in obese subjects, affects 17 million adults in the United States, with a continuous and alarmingly increasing rate. To prevent development of diabetes in those who are at high risk, it is recommended to optimize meal planning and enhance physical activity to make sustained weight reduction possible. In addition to lifestyle changes, various oral antidiabetic agents are available, with diverse mechanisms of action. Some target defective insulin secretion (sulphonylureas, benzoic acid derivatives) or glucose absorption (glycosidase inhibitors), whereas others target insulin resistance (metformin, thiazolidinediones). Patients with metabolic syndrome and diabetes have an increased risk for cardiovascular disease linked to a higher prevalence of hypertension, dyslipidemia, microalbuminuria, and altered hemostasis--parameters that may be modified by antidiabetic agents. In this article, we review the oral agents used to treat type 2 diabetes and the metabolic syndrome, and their effects on vascular tissue. PMID- 15748535 TI - ACE inhibitors in patients with vascular disease: should the PEACE trial change medical practice? PMID- 15748536 TI - BENEDICT in the treatment of hypertension. PMID- 15748537 TI - Is angiotensin II made inside or outside of the cell? AB - Angiotensin synthesis at tissue sites is well-established, and depends largely, if not completely, on kidney-derived renin. The exact tissue site of angiotensin generation (extracellular fluid, cell surface, intracellular compartment) is still being debated. In this review, we discuss the various possibilities, taking into consideration the intracellular occurrence/absence of prorenin, renin, angiotensinogen, angiotensin-converting enzyme, and angiotensin receptors; the local activation of prorenin to renin; the differences between in vivo and in vitro studies; and the methodologic difficulties related to angiotensin measurements. It is eventually concluded that angiotensin generation at tissue sites occurs extracellularly, most likely on the cell surface. PMID- 15748538 TI - Role of angiotensin III in hypertension. AB - The hyperactivity of the brain renin-angiotensin system (RAS) has been implicated in the development and maintenance of hypertension in several types of experimental and genetic hypertension animal models. Among the main bioactive peptides of the brain RAS, angiotensin (Ang) II and Ang III display the same affinity for type 1 and type 2 Ang II receptors. Both peptides, injected intracerebroventricularly, similarly increase blood pressure (BP); however, because Ang II is converted in vivo to Ang III, the identity of the true effector is unknown. In this article, we review new insights into the predominant role of brain Ang III in the control of BP, underlining the fact that brain aminopeptidase A (APA), the enzyme-forming central Ang III, could constitute a putative central therapeutic target for the treatment of hypertension. This justifies the development of potent systemically active APA inhibitors, such as RB150, as prototypes of a new class of antihypertensive agents for the treatment of certain forms of hypertension. PMID- 15748539 TI - Molecular evidence of tissue renin-angiotensin systems: a focus on the brain. AB - Hypertension remains one of the largest human health problems, because hypertensive patients carry increased risk for ischemic heart disease, stroke, atherosclerosis, and renal failure. The renin-angiotensin system (RAS) has been intensively investigated for more than 100 years because it is a powerful regulator of blood pressure, and the antihypertensive benefits of RAS inhibitors are very clear. Despite a wealth of clinical and basic studies, the precise mechanisms by which the RAS regulates blood pressure remains incomplete. In this chapter, we review data demonstrating the existence and function of intrinsic tissue RAS, with a primary focus on the brain. PMID- 15748543 TI - Omalizumab: a recombinant humanized monoclonal IgE-blocking antibody. AB - Omalizumab (Xolair) is a recombinant humanized monoclonal antibody that blocks the high-affinity Fc receptor of immunoglobulin E (IgE). It is used to treat patients with moderate-persistent to severe-persistent asthma; patients must be older than 12 years, have a positive skin test to a perennial aeroallergen (e.g., dust mites, cats, dogs, and mold), and be symptomatic with inhaled corticosteroids. Omalizumab has a low incidence of side effects, and it is very expensive. The exact dosage of omalizumab dosage is determined by body weight and pretreatment serum total IgE levels. Omalizumab may have a role in the treatment of atopic dermatitis when IgE plays a causal role. PMID- 15748542 TI - Treatments for mild-to-moderate recalcitrant plaque psoriasis: expected clinical and economic outcomes for first-line and second-line care. AB - The cost effectiveness of treatments for psoriasis has been evaluated previously by several different investigators. Such evaluations should be updated as new products or data become available. To this end, a comparison of expected treatment-related clinical and economic outcomes is undertaken from the payer perspective using a disease-intervention model, decision analyses, and newly emergent information. The model is based on academy guidelines and recommended clinical practice. Model inputs (clinical and cost data) are culled from the medical literature and advisory clinical assessment surveys. Comparable therapies are various topical pharmacotherapies and phototherapies, including the 308-nm excimer laser (XTRAC, PhotoMedex, Montgomeryville, PA). Analytic results indicate that clinical and economic outcomes are influenced by treatment selections but are muted by the rotational nature of treatment regimens. Multiple analyses are required to reveal individual product performance. On the basis of these analyses, the addition of the 308-nm excimer laser to the rotational mix of treatments commonly utilized as second-line therapies for mild-to-moderate plaque psoriasis is expected to add incremental clinical benefit for patients without incremental cost for payers, because the laser can replace both more costly and less costly alternatives for appropriately selected patients who require a different therapeutic modality to maintain or improve their responsiveness. PMID- 15748541 TI - Cellular mechanisms and treatment of diabetes vascular complications converge on reactive oxygen species. AB - High glucose activates a myriad of signaling and gene expression pathways in non insulin-dependent target cells causing diabetes complications. One of the earliest responses to high glucose by vascular cells is the generation of reactive oxygen species (ROS) that act directly on intracellular proteins and DNA, or indirectly as second messengers, transforming these cells into disease phenotypes. ROS are produced by mitochondria and/or NADPH oxidase in all target cells exposed to high glucose studied to date. Reports using cell cultures and diabetic animal models indicate that inhibition of ROS generation prevents the amplification of signaling and gene expression that are implicated in vascular complications. These models convincingly demonstrate that maneuvers preventing ROS production attenuate or completely abrogate early micro- and macrovascular end-organ damage of diabetes, including nephropathy, retinopathy, and large vessel atherosclerosis. Attention now turns to the development of more effective antioxidants that could be used in clinical trials in the prevention and treatment of diabetes complications. PMID- 15748540 TI - The salt paradox and its possible implications in managing hypertensive diabetic patients. AB - Diabetes mellitus is one of the leading causes of end-stage renal disease. The pathogenesis of diabetic nephropathy is still poorly understood, but glomerular injury has been ascribed, at least in part, to glomerular hyperfiltration, which occurs early in the course of diabetes mellitus. Therefore, a better understanding of the early dysfunctions observed in the diabetic kidney may permit the development of new strategies to prevent diabetic nephropathy. In this review, we discuss the pathophysiology for the paradoxical relationship between dietary salt and glomerular filtration rate observed in early diabetes mellitus and possible implications in managing diabetic patients. PMID- 15748544 TI - The potential immunomodulatory effects of topical retinoids. AB - New research has refined our understanding of the immunopathophysiology of acne. Various immune factors, including both innate and adaptive immune responses, have been implicated in the pathophysiology of inflammatory acne. Topical retinoids such as tretinoin, adapalene, and tazarotene, exhibit immunomodulatory effects that may help to explain their efficacy in the resolution of inflammatory lesions. PMID- 15748545 TI - Cutaneous leishmaniasis in Pakistan. AB - Leishmaniasis is a major health problem worldwide. It is also a particular problem in the rural areas of Pakistan. The disease occurs in varying presentations, from the self-limited and even self-healing cutaneous forms to fatal systemic disease. Lesions of cutaneous leishmaniasis may occur anywhere on the body but the most likely sites are the exposed parts. The initial papule rapidly gives rise to an ulcer. Systemic leishmaniasis is rarer in Pakistan and invariably fatal if not treated promptly. It affects the internal body organs, particularly the spleen and the liver. Leishmaniasis is transmitted by an infected female sandfly. Cutaneous lesions are usually single and often self healing, but a presentation with multiple ulcers resulting from multiple bites from the sandfly is not rare in Pakistan. The disease has a very long history and lesions like leishmaniasis have been described dating back to the ninth century (Balkan sore). Cutaneous leishmaniasis has been given various names in different civilizations such as "Delhi boil" in India, "Baghdad boil" in Iraq, and "saldana" in Afghanistan. The organism responsible for leishmaniasis was discovered 100 years ago but the disease has not been eradicated; rather it is on rise in many parts of the world. If control measures are not taken, it might emerge as a major health problem. Pakistan has a burden of cutaneous and visceral leishmaniasis, the mucocutaneous form being almost nonexistent. The physicians need to know the diagnostic criteria as well as the treatment of the disease. Because of a scarcity of dermatologists in the rural areas, most of these cases present to general practitioners. Control of this disease is further complicated by an inadequate supply of appropriate drugs. PMID- 15748546 TI - Primary, systemic amyloidosis and the dermatologist: where classic skin lesions may provide the clue for early diagnosis. AB - Primary amyloidosis may present with skin lesions as a primary or sole expression of underlying plasma-cell dyscrasia. Classic skin lesions of primary, systemic amyloidosis are listed, and features suggestive of the diagnosis are discussed. Where this condition is considered in the dermatologist's differential, the investigations described may lead to an early diagnosis. When therapy can be initiated before the onset of organ failure, survival may be prolonged. PMID- 15748547 TI - Dermatitis herpetiformis and gluten-free diet. PMID- 15748548 TI - Punch biopsies of melanoma: a diagnostic peril. AB - Excisional or incisional biopsies of melanoma are used to determine depth of tumor invasion and to plan subsequent treatment. Accurate determination of depth of melanoma invasion is critical for treatment decisions and prognosis. Incisional or punch biopsies can be perilous for histopathologic determination of invasion, and both over- and underestimation of invasion can occur when using incisional methods. Likewise, histologic factors can lead to over- and underestimation of invasion. Prognosis and treatment of melanoma are primarily determined by depth of tumor invasion. We discuss several scenarios that can lead to over- and underestimation of depth of invasion in incisional biopsy specimens. We therefore discourage incisional or punch biopsies of suspected melanoma and recommend that depth of invasion not be reported on these types of specimens. PMID- 15748549 TI - Medical student dermatology research in the United States. AB - The development of academic researchers is important for the future success of dermatology, but few dermatology trainees are entering academic practice in the United States. Because many successful researchers begin to focus on research skills during medical school, an electronic survey was conducted to evaluate dermatology research opportunities and benchmarks for U.S. medical students. First- and second-year medical students participated in case studies, clinical research projects, and laboratory benchwork, but participation significantly increased during the third and fourth years. Funded programs had higher student participation than nonfunded programs and were also more likely to have medical students present their research at meetings. A wide variety of research opportunities are currently offered to U.S. medical students, so a lack of research opportunities in medical school does not explain the dwindling supply of academic dermatologists in the United States. Because funding appears to be an important factor for successful medical student research, new mechanisms of funding should be identified and developed. PMID- 15748550 TI - The use of apraclonidine eyedrops to treat ptosis after the administration of botulinum toxin to the upper face. AB - A side effect of the injection of botulinum toxin into the upper third of the face is ptosis or lid droop. A therapy recommended to treat ptosis resulting from administration of botulinum toxins A and B is Iopidine (apraclonidine 0.5 %) eye drops. Apraclonidine is an alpha2-adrenergic agonist, which causes Muller muscles to contract quickly elevating the upper eyelid 1-3 mm. Little published data discusses the use of apraclonidine to treat such ptosis. This communication discusses the extant literature on this usage. Research needs to be done to establish the utility and dosing of apraclonidine for botulinum toxin-induced ptosis. PMID- 15748551 TI - Modified Burow's advancement flaps for two circular adjoining cutaneous defects: the "8"-to-"S"-plasty. AB - The traditional method to close two closely approximated cutaneous defects (i.e., a figure "8"-shaped defect) sacrifices healthy skin and does not give optimal aesthetic results. We report a case with modified Burow's advancement flaps describing less sacrificed skin and shorter scars. Modified Burow's advancement flaps ("8"-to-"S" plasty) were applied, sparing two Burow skin triangles. Application of that modified operation technique leads to shorter scars in comparison to traditional Burow's advancement flaps. A modified technique is presented here for skin defects shaped like an "8"--namely, two similar round and adjoining lesions. The suture line after closing resembles an "S". With this 8-to S plasty no additional incisions are necessary. PMID- 15748552 TI - Burow's advancement flap closure of adjacent defects. AB - When two surgical defects are closely approximated, primary closure may be difficult because of tension on the tissue between the defects. We outline a technique using a Burow's-triangle advancement flap in which the advanced Burow's triangle contains the second defect. The defects are easily closed with a single flap that utilized the second defect. This flap is useful when there are two closely approximated surgical defects of which primary closure is limited by tension on the tissue between the defects. PMID- 15748553 TI - Systemic argyria associated with ingestion of colloidal silver. PMID- 15748554 TI - Pemphigus vegetans in a patient with lung cancer. AB - A 66-year-old woman with a history of lung cancer treated with radiotherapy 2 years prior to admission, was seen for mucosal and skin lesions of 3-month's duration. She had pustules involving intertriginous areas and erosions involving the oral mucosa. Histopathology of skin lesions and the results of direct and indirect immunofluorescence studies were consistent with the Hallopeau-type pemphigus vegetans. Additionally, circulating antibodies against skeletal muscles were detected in patient's serum. The patient was treated with immunosuppressive therapy and had an almost complete remission of skin and mucosal lesions within 1 month; however, the patient developed pneumonia with pyothorax, apparently related to recurrence of lung cancer. PMID- 15748555 TI - Tattoo pigment in sentinel lymph nodes: a mimicker of metastatic malignant melanoma. AB - Tattoo pigment in the sentinel lymph nodes of melanoma patients represents a clinical challenge. If a tattoo is present in the area of the primary melanoma, the draining lymph nodes are likely to contain tattoo pigment, as well as being the site for metastatic deposits of melanoma. We describe a case report involving an elderly Caucasian male diagnosed with a Clark level-4 nodular malignant melanoma, wherein intraoperatively we encounter a darkly pigmented lymph node highly suspicious for metastatic disease. The patient had a tattoo in the vicinity of the malignant melanoma The specimen is sent for histological examination and is found to contain pigmented macrophages, but metastatic malignant melanoma is not identified. Histological confirmation of an enlarged pigmented node is essential before radical surgery is performed. PMID- 15748556 TI - Apert syndrome. PMID- 15748557 TI - A case with widespread cutaneous metastases of unknown primary origin: grave prognostic finding in cancer. AB - We present a 52-year-old man with widespread cutaneous metastases (CMs) of unknown primary origin. Although we performed many of the investigations, we could not find out a primary origin of malignancy. There are no practical algorithms to identify the primary of cutaneous metastatic tumors of unknown origin. An algorithm in cancer patients with CMs seems to be needed to manipulate such cases. We believe that more reports related with this issue must be published to form an algorithm in such cases. PMID- 15748558 TI - Collagen XVIII and fibronectin involvement in bullous scleroderma. AB - BACKGROUND: Endostatin, an anti-angiogenic C-terminal fragment of collagen XVIII, has been recently reported to play a role in scleroderma pathogenesis, but collagen XVIII immunohistochemistry in scleroderma skin has still not been performed. Bullous scleroderma, a rare form of scleroderma, may have altered angiogenic and lymphangiogenic characteristics. OBJECTIVE: Our aim is to report a rare case of bullous scleroderma, studying the presence of fibronectin and collagens type I, III and XVIII in sclerodermic skin. METHODS: We describe the progression of bullous scleroderma in a 67-year-old patient since the first symptoms. Histological and immunohistochemical aspects of skin biopsies are compared to normal skin from a patient without scleroderma and are correlated with the pathogenesis of the disease. Indirect immunofluorescence measured by laser confocal microscopy allows quantitative determination of fibronectin and collagens type I, III and XVIII. RESULTS AND CONCLUSIONS: Dermo-epidermal cleavage, fibrosis and inflammation are the main histological findings. The dermal distribution and amounts of collagens and in the scleroderma patient are similar to normal skin. Conversely, both fibronectin and collagen XVIII are increased in scleroderma skin, suggesting their involvement in the pathogenesis of bullous scleroderma. PMID- 15748559 TI - Seborrheic keratosis-like porokeratosis: a case report. PMID- 15748560 TI - Disseminated cutaneous rhinosporidiosis. AB - Rhinosporidiosis is a chronic granulomatous disorder caused by Rhinosporidium seeberi. It frequently involves the nasopharynx and occasionally affects the skin. We herewith report a 55-year-old man who has disseminated cutaneous rhinosporidiosis. He presents with multiple reddish lesions over the nose of 10 year's duration. In the past year, he develops skin lesions over the right arm and over back. Histopathological examination of the skin biopsy specimen from the representative cutaneous lesions shows hyperplastic epithelium with numerous globular cysts of varying shape, representing sporangia in different stages of development. His serology for HIV infection by ELISA is negative. On the basis of these clinical and histopathological findings, a diagnosis of nasal rhinosporidiosis with cutaneous dissemination is made. PMID- 15748561 TI - Genital leiomyoma: surgical excision for both diagnosis and treatment of a unilateral leiomyoma of the male nipple. AB - We describe a rare case of unilateral leiomyoma of the nipple in a man presenting with pruritus of the nipple for 3 months. A conservative surgical excision is performed for diagnosis. Histologic examination and immunostaining confirmed the diagnosis of leiomyoma. The leiomyoma is completely excised, and the patient denies pruritus after surgery. Conservative surgical excision of a persistently hard and pruritic nipple can be effectively used as both diagnostic measure and treatment modality. PMID- 15748562 TI - Multiple cutaneous hemangiomas accompanied by hepatic hemangiomas. PMID- 15748563 TI - Localized forms of steatocystoma multiplex: case report and review of the literature. AB - A 70-year-old man presents with multiple asymptomatic, yellow to skin-colored firm papules scattered on the scalp; the lesions had been present for about 30 years. Histological findings are consistent with the diagnosis of steatocystoma. We review the English-language literature of the published cases of localized forms of steatocystoma multiplex. PMID- 15748564 TI - Syphilis presenting as erosive papules on the palate. PMID- 15748565 TI - Cutaneous metastases from carcinoma of the prostate: a case report. PMID- 15748566 TI - Stevens-Johnson syndrome associated with single high dose of lamotrigine in a patient taking valproate. PMID- 15748568 TI - Pityriasis rosea with palmoplantar plaque lesions. AB - Pityriasis rosea is a skin disease characterized by sharply defined pruritic red patches covered by fine scales. It affects mostly adolescent and young adults. Typical lesions usually affect the trunk in a Christmas-tree pattern. The eruption usually resolves after 6 weeks but symptomatic treatment may be needed. Two patients are reported with classic presentation of pityriasis rosea except for the unusual associated palmoplantar lesions; both patients had negative RPR (with dilutions) and MHA-TP. They responded to 2-week courses of either oral erythromycin or Clarithromycin with complete resolution. PMID- 15748567 TI - Primary epidermoid carcinoma of the submandibular salivary gland. PMID- 15748569 TI - Videodermatoscopy: a useful tool for diagnosing cutaneous dystrophic calcifications. AB - A suspected diagnosis of cutaneous calcification is based on the presence of slightly elevated or ulcerated firm, white papules and plaques; these lesions often extrude white, chalky material. In clinical practice, however, a biopsy is required to confirm clinical suspicions. We attempted to demonstrate cutaneous calcinosis in vivo by using Raman spectroscopy, a nondestructive analytical method based on the analysis of light scattered from the investigated material. We used digital videodermatoscopy to improve the detection of cutaneous calcifications in vivo in two patients with CREST syndrome. A characteristic flower-like arrangement was observed at x-50 magnification. We believe this pattern may be highly indicative of cutaneous dystrophic calcinosis. PMID- 15748570 TI - Clinical and histopathological improvement of psoriasis with oral chondroitin sulfate: a serendipitous finding. AB - We describe the clinical and histopathological results of plaque psoriasis in eleven adult patients with knee osteoarthritis and long-standing, moderate to severe psoriasis resistant to conventional therapy treated with chondroitin sulfate. Patients received 800 mg per day of chondroitin sulfate for 2 months. Skin biopsies were obtained before and after treatment. All patients but one presented a dramatic improvement of the condition of the skin, with a reduction of swelling, redness, flaking, and itching (clearance of psoriasis in one patient), increase in the hydration and softening of the skin, and amelioration of scaling. Histopathologically, there was a statistically significant decrease in epidermal thickness, a decrease in the thickness between the stratum basale and the stratum granulosum, a significant improvement of the degree of psoriasis activity, and a decrease in the number of keratinocytes stained with Ki-67. The confirmation of these serendipitous findings in controlled prospective studies could represent an important advance in the therapeutic armamentarium for patients with psoriasis given the excellent safety profile of chondroitin sulfate. PMID- 15748571 TI - Telangiectasia macularis eruptiva perstans. AB - A 45-year-old man presented with lesions clinically and histologically consistent with telangiectasis macularis eruptiva perstans (TMEP). TMEP is a rare form of mastocytosis, which is localized to the skin, although systemic involvement has been reported. A mutation in the c-kit proto-oncogene is thought to be involved in mast cell hyperplasia in some patients with mastocytosis. Treatment depends on the presence of systemic involvement or clinical symptoms. In case reports, therapeutic benefit has been reported in TMEP with the use of the 585-flashlamp pumped dye lasers and of electron beam radiation. PMID- 15748572 TI - Rubinstein-Taybi syndrome (broad thumb-hallux syndrome). AB - Rubinstein-Taybi syndrome is a rare genodermatosis with characteristic features that include downward sloping palphebral fissures, broad thumbs and halluces, and mental retardation. Dermatologic manifestations include capillary malformations, keloid formation, and pilomatricomas. Systemic features may involve the cardiac, audiologic, ophthalmologic, endocrine, neurologic, and respiratory systems. The syndrome is sporadic in nature and has been linked to microdeletion at 16p13.3 encoding CREB-binding protein gene (CREBBP). PMID- 15748574 TI - Hyperimmunoglobulin E syndrome. AB - A 63-year-old man presented with erythroderma, peripheral blood eosinophilia, elevated serum IgE levels, and a history of dermatitis, furunculosis, and a cold abscess. Hyperimmunoglobulin E syndrome is a rare, multisystem disorder that is characterized by cutaneous and sinopulmonary infections, dermatitis, and elevated serum IgE levels. Traditional therapy includes good skin care, antibiotics for skin and pulmonary infections, incision and drainage of abscesses, and emollients plus topical glucocorticoids for dermatitis. PMID- 15748573 TI - Cowden's syndrome. AB - A 59-year-old woman initially presented with facial papules, which were consistent with trichilemmomas on histopathologic examination. Her course was complicated by breast, endometrial, and renal-cell carcinomas, as well as a multinodular goiter that necessitated a thyroidectomy. Cowden's syndrome is a rare autosomal dominant syndrome that is characterized by hamartomas of ectodermal, mesodermal, and endodermal origin as well as an increased risk of breast, thyroid, and endometrial neoplasias. Medical management includes screening for breast cancer and thyroid abnormalities, and performing other age specific examinations. Treatment of facial papules includes topical 5 fluorouracil, isotretinoin, curettage, laser ablation, or surgical excision. PMID- 15748575 TI - Familial leiomyomatosis cutis et uteri. AB - A 45-year-old woman presented with multiple, small, asymptomatic, hyperpigmented to skin-colored, smooth, dermal papules on the right temple as well as with uterine fibroids. She has a family history of uterine fibroids and cutaneous leiomyomas. An autosomal dominant disorder of multiple cutaneous leiomyomas and uterine fibroids (Reed syndrome) has been localized to a gene on chromosome 1q42.3-43. This gene encodes fumarate hydratase, which is an enzyme in the Kreb cycle, that acts as a tumor suppressor in this familial disorder. A subset of people may be at risk for papillary renal cell carcinoma. PMID- 15748576 TI - Papulonodular mucinosis with subacute cutaneous lupus erythematosus. AB - A 30-year-old woman with subacute cutaneous lupus erythematosus presented with a facial eruption, painful and swollen ulcers of her digits, and asymptomatic lesions on her abdomen. The laboratory findings were consistent with subacute cutaneous lupus erythematosus with an anticardiolipin antibody. A biopsy specimen of the asymptomatic lesions on her abdomen showed mucin within the upper reticular dermis. Papulonodular mucinosis is a rare but well-documented finding associated with systemic and cutaneous forms of lupus erythematosus. The etiology and pathogenesis of this finding remain to be elucidated. PMID- 15748577 TI - Dermatomyositis. AB - A 51-year-old woman presented with weakness in her arms and legs and an eruption on the dorsal aspects of the hands, upper back, and face. Histopathologic features showed vacuolar alteration of the basal layer, a thick basement membrane, and deposits of connective-tissue mucin in the papillary dermis and the upper reticular dermis. Dermatomyositis is an idiopathic disease that is characterized by specific cutaneous manifestations and myopathy, which may be associated with occult malignancy. Treatment options include corticosteroids and other immunosuppressive agents such as methotrexate, cyclosporine, mycophenolate mofetil, and azathioprine. PMID- 15748578 TI - Pityriasis lichenoides chronica. AB - A 19-year-old woman presented with a five-year history of guttate macules and yellow to skin-colored papules with collarette of fine scale on the trunk and the upper and lower extremities. Guttate pityriasis lichenoides chronica is an uncommon presentation of this T-cell-mediated disease. It is of unknown etiology; however, an infectious agent has been suspected. It is considered in a spectrum with pityriasis lichenoides et varioloformis acuta and rarely mycosis fungoides and CD30 lymphomas. Improvement has been shown after treatment with oral tetracyclines, ultraviolet B and UVA1 phototherapy, and PUVA photochemotherapy. PMID- 15748579 TI - Acrodermatitis continua. AB - A 54-year-old woman presented with a 5-year history of erythema and pustules on the distal portion of her left index finger. Acrodermatitis continua of Hallopeau is a rare, chronic, sterile pustular eruption affecting the distal aspects of the digits. It is often considered to be a variant of pustular psoriasis that tends to be resistant to both topical and systemic treatments for psoriasis. PMID- 15748580 TI - Granulomatous cheilitis. AB - A 54-year-old man presented with lip swelling, which was not altered by a trial of medication discontinuation. Patch tests showed reactions to bromonitropropane and gold, which were not clinically relevant. Treatment included topical and intralesional glucorticoids as well as tacrolimus ointment. Granulomatous chelitis an idiopathic disorder that is characterized by painless lip edema. It may be found as part of the triad of Melkersson-Rosenthal syndrome. Treatment includes intralesional glucocorticoids as well as systemic therapies. PMID- 15748581 TI - Carney complex. AB - A 55-year-old man with a history of two atrial myxomas presented for evaluation and removal of skin lesions. Histopathologic examination showed a well circumscribed lesion that was consistent with a myxomatous polyp. Carney complex is a syndrome with multiple neoplasias, which is characterized by mucocutaneous myxomas, cardiac myxomas, lentigines, and endocrine abnormalities. Patients with Carney complex need to be monitored for the development of cardiac myxomas and endocrine tumors. PMID- 15748582 TI - Cutaneous Rosai-Dorfman disease. AB - Presented is a case of a 31-year-old woman with cutaneous Rosai-Dorfman disease, which is a rare, benign, generally self-limited histiocytic proliferative disorder that is characterized by painless lymphadenopathy, fever, neutrophilia, an elevated erythrocyte sedimentation rate, and polyclonal hypergammaglobulinemia. It primarily involves lymph nodes but can affect extranodal sites such as the skin, where it typically appears as indistinct erythematous papules and nodules. On histopathologic examination, emperipolesis is a consistent finding. The etiology is unknown, and treatment is based on clinical manifestations. PMID- 15748583 TI - Primary (idiopathic) cold urticaria and cholinergic urticaria. AB - A 76-year-old man with a longstanding history of cold sensitivity developed wheals after the application of an ice cube. Cold urticaria is a type of physical urticaria that is characterized urticaria and angioedema after exposure to cold. It may be idiopathic or secondary to hematologic or infectious diseases. Treatment of primary cold urticaria includes antihistamines; however, ketotifen, doxantrazole, zafirlukast, cyclosporine, and cold-tolerance induction may be tried in refractory cases. PMID- 15748584 TI - Behcet's disease. AB - A 29-year-old man presented with oral and genital ulcers, erythematous papules and pustules on his back and chest, and deep vein thrombi. A diagnosis of Behcet disease was made. Behcet disease is a relapsing disorder that affects the mucocutaneous surfaces. It presents usually as ulcers on the orogenital mucosae, but can also present as an acneiform eruption. The International Study Group on Behcet disease has established criteria that consist of oral and genital lesions, ocular involvement, skin findings, and a pathergy. Treatment of choice is colchicine or prednisone. PMID- 15748585 TI - Birt-Hogg-Dube syndrome. AB - 27-year-old woman presented with multiple growths on her face and upper body, which she has had for several years. She also reported that her mother and maternal grandmother have similar lesions and that her mother has been diagnosed with Birt-Hogg-Dube syndrome. Histopathologic features of one papule demonstrated basaloid cells emanating from a dilated follicular infundibula surrounded by a well-circumscribed fibromucinous stroma. Birt-Hogg-Dube syndrome is an autosomal dominant-inherited condition characterized by fibrofolliculomas, trichodiscomas, and acrochordons. It is associated with renal tumors, spontaneous pneumothoraces, and possibly intestinal polyps. Treatment includes evaluation for associated renal, lung, and intestinal findings as well as cosmetic removal of skin lesions. PMID- 15748586 TI - Acquired perforating disorder. AB - A 53-year-old man presented with asymptomatic, hyperkeratotic papules and hyperpigmented macules of the face and upper extremities. Treatment was initiated with topical tretinoin and intralesional glucocorticoids. Acquired perforating disorder is a disorder of keratinization that is associated with diabetes mellitus and uremia. Classic presentation involves hyperkeratotic papules with a central core, which occur most frequently on the lower extremities. Treatment includes topical tretinoin, glucocorticoids, keratolytics, systemic retinoids and glucocorticoids, and ultraviolet B phototherapy. PMID- 15748587 TI - Tuberous sclerosis complex (Bourneville disease). AB - A 26-year-old woman with tuberous sclerosis complex had numerous cutaneous manifestations and systemic manifestations that included subependymal nodules, pulmonary lymphangioleiomyomatosis, renal cysts, and bilateral renal angiomyolipomas. Tuberous sclerosis is a hamartomatous disease with defects in either hamartin (TSC1) or tuberin (TSC2) that can be of autosomal dominant inheritance or spontaneous mutation. PMID- 15748588 TI - Necrobiosis lipoidica. AB - A 68-year-old woman presented with a 3-year history of asymptomatic, yellow-brown plaques on the lower extremities with trauma-related superficial ulcers. A biopsy specimen was consistent with necrobiosis lipoidica. Necrobiosis lipoidica is a chronic granulomatous dermatitis that is associated with diabetes mellitus; however, its pathogenesis remains unclear. Topical glucocorticoids are first-line therapy; however, no treatment has proven efficacy in double-blind, placebo controlled studies. PMID- 15748589 TI - Multiple eruptive keratoacanthomas, de novo. AB - An 89-year-old man presented with the sudden eruption of multiple, erythematous papules and nodules on his lower extremities. A skin biopsy specimen showed keratoacanthoma. After 3 months of treatment with topical imiquimod cream, the keratoacanthomas failed to resolve. We review the different clinical subtypes, histologic features, and treatment options for multiple eruptive keratoacanthomas. PMID- 15748590 TI - Axillary granular parakeratosis. AB - A 71-year-old man developed a pruritic axillary eruption. Histopathologic examination showed laminated orthokeratosis, parakeratosis, and hypergranulosis. There changes were consistent with a diagnosis of axillary granular parakeratosis. Axillary granular parakeratosis is an intertriginous eruption that is usually found in the axillae of middle-aged women and is characterized clinically by pruritic, erythematous, hyperkeratotic plaques and histologically by parakeratosis with retention of keratohyaline granules. Pathophysiology is thought to involve a defective profilaggrin-filaggrin pathway. Evidence-based treatment of this disorder is not available. PMID- 15748591 TI - Mycobacterium marinum infection. AB - A 49-year-old man presented with nodules on his right hand after a history of Mycobacterium marinum infection recently treated with rifampin and clarithromycin. The patient has an aquarium with Betta fish (Siamese fighting fish). PMID- 15748592 TI - Follicular mucinosis associated with mycosis fungoides. AB - A 62-year-old man with a 13-year history of mycosis fungoides presented with a 2 month history of alopecia of the scalp. The mycosis fungoides had remained untreated for the previous 3.5 years. A biopsy specimen from the scalp showed follicular mucinosis in association with mycosis fungoides. PMID- 15748593 TI - Rapp-Hodgkin syndrome. AB - A 5-year-old boy with a history of a bifid uvula and a submucosal cleft palate presented for evaluation of brittle nails. The physical examination demonstrated cup-shaped ears, a broad nasal root, thin upper lip, mid-facial hypoplasia, coarse hair, and twenty-nail dystrophy. The clinical presentation of ectodermal dysplasia with cleft palate was consistent with Rapp-Hodgkin syndrome, which is one of several allelic diseases associated with mutations in the TP63 gene. The clinical manifestations of Rapp-Hodgkin as well as other ectodermal dysplasias with clefting are discussed. PMID- 15748594 TI - Sarcoidosis. AB - A 48-year-old woman with an 8-year history of intermittently pruritic papules on the lower extremities suddenly developed pruritus and generalized spread of the lesions over a 1-month period. The lesions appeared as smooth, flat-topped, violaceous, round papules on all extremities, trunk, and back. A biopsy specimen showed sarcoidosis. Cutaneous sarcoidosis has many morphologic presentations and often mimics other dermatologic diseases. Despite widespread cutaneous involvement, she has no systemic involvement. Treatment options are reviewed. PMID- 15748595 TI - Mitochondrial DNA analysis in clinical laboratory diagnostics. AB - Mitochondrial disorders are increasingly being diagnosed, especially among patients with multiple, seemingly unrelated, neuromuscular and multi-sytem disorders. The genetics are complex, in particular as the primary mutation can be either on the nuclear or the mitochondrial DNA (mtDNA). mtDNA mutations are often maternally inherited, but can be sporadic or secondary to autosomally inherited mutations in nuclear genes that regulate mtDNA biosynthesis. mtDNA mutations demonstrate extreme variable expressivity in terms of clinical manifestations and severity, even within a family. Disease is often episodic. Several well-defined clinical syndromes associated with specific mutations are described, yet the genotype-phenotype correlation is fair at best and most patients do not fit within any defined syndrome and have rare or novel mutations. In most patients, mutant and wild-type mtDNA coexist ("heteroplasmy"), although homoplasmic mtDNA mutations also are known. "Standard" mtDNA clinical diagnostics usually consists of a PCR-based assay to detect a small number of relatively common point mutations and Southern blotting (or PCR) for large (>500 bp) rearrangements. In selected cases testing negative, additional analyses can include real-time PCR for mtDNA depletion, and full mtDNA genome screening for the detection of rare and novel point mutations by a variety of methods. Prenatal diagnosis is problematic in most cases. PMID- 15748596 TI - Circulating cardiovascular markers and mediators in acute illness: an update. AB - An update is given of the circulating markers and mediators of cardiovascular dysfunction in acute illness. Some of these circulating markers reflect mediator action on the peripheral vasculature, such as endothelium-derived endothelin and nitrite/nitritate, the stable end products of nitric oxide. Other markers mainly reflect actions on the heart, such as the natriuretic peptide family, released from the heart upon dilatation, serving as a marker of congestive heart failure and potentially having negative inotropic effects. Indeed, some factors may be both markers as well as mediators of cardiovascular dysfunction of the acutely ill and bear prognostic significance. Assessing circulating levels may help refine clinical judgment of the cardiovascular derangements encountered at the bedside, together with clinical signs and hemodynamic variables. For instance, assessing natriuretic peptides in patients with pulmonary edema of unclear origin may help to diagnose congestive heart failure and cardiogenic pulmonary edema, when the pulmonary capillary wedge pressure is not measured or inconclusive. Future aligning of hemodynamic abnormalities with patterns of circulating cardiovascular markers/mediators may help to stratify patients for inclusion in studies to assess the causes, response to therapy and prognosis of cardiovascular derangements in the acutely ill. PMID- 15748597 TI - Autoantibodies in Type 1 diabetes. AB - BACKGROUND: Autoimmune diseases are associated with autoantibodies which recognise specific antigens. METHODS: Autoantibodies can be detected by various methods including immunofluorescence and radioimmunoassays using disease-specific radiolabelled recombinant autoantigens As recombinant assay have high-throughput potential, it is now even possible to screen whole populations. RESULTS: Diabetes associated autoantibodies including autoantibodies to glutamic acid decarboxylase are valuable in disease classification and disease prediction. Some autoantibodies are more predictive of diabetes than others. CONCLUSIONS: Autoantibodies, and the antigens they recognise, are potentially valuable in the prediction and prevention respectively of type 1 diabetes as well as in the classification of autoimmune diabetes. Screening the general population for type 1 diabetes susceptibility is now feasible. Trials to prevent type 1 diabetes using disease-associated autoantigens are now in progress. PMID- 15748598 TI - A sensitive assay of red blood cell sorbitol level by high performance liquid chromatography: potential for diagnostic evaluation of diabetes. AB - BACKGROUND: The level of sorbitol within erythrocyte or other tissue is increased during diabetic complications such as neuropathy, retinopathy, and nephropathy and is thus considered as an indicator for the diagnosis and monitoring the diabetic status. A highly sensitive method of estimation of RBC sorbitol may be very useful in determining the state of diabetes. In this study a high performance liquid chromatography method was developed to analyze RBC sorbitol level. METHOD: Benzoylation was used for the precolumn derivatization of RBC polyol prior to analyze by HPLC using C-18 reversed phase column and UV detector. RESULTS: The sensitivity was 5 ng/ml using standard sorbitol. The linearity was highly satisfactory up to 2.5 mug/ml which was adequate to measure RBC sorbitol in both diabetic and normal subjects. The values were significantly higher for the diabetic patient compared to normal. Within-run assay CVs were <10% and <14% for between-run. CONCLUSION: This method could be useful in estimating RBC sorbitol content for the clinical evaluation of diabetic patients. PMID- 15748600 TI - Impairment of sodium pump and Na/H exchanger in erythrocytes from non-insulin dependent diabetes mellitus patients: effect of tea catechins. AB - BACKGROUND: Tea catechins (EGCG, EGC, ECG and EC) possess many important biological properties. We evaluated the effect of tea catechins on erythrocyte membrane Na(+)/K(+)-ATPase and sodium/hydrogen exchanger (NHE) activity in normal (control) and NIDDM subjects. METHODS: Erythrocyte membrane Na(+)/K(+)-ATPase and NHE activity were determined in normal and non-insulin dependent diabetes mellitus (NIDDM) patients. In vitro effect of tea catechins was studied by incubating membrane/intact erythrocytes in assay medium prior to Na(+)/K(+) ATPase/NHE activity determination. RESULTS: A 24.2% decrease in the activity of Na(+)/K(+)-ATPase (p<0.001) and 39.37% increase in activity of NHE (p<0.02) were observed in NIDDM subjects compared to normal. Tea catechins inhibited the activity of Na(+)/K(+)-ATPase and NHE in both normal and NIDDM erythrocytes, the effect was concentration-dependent. The inhibitory effect of EGCG and ECG at micromolar concentrations was greater compared to EGC and EC on Na(+)/K(+) ATPase. On NHE the inhibition of tea catechins was in the order: EC>EGC>ECG>EGCG at concentrations up to 10 micromol/l. CONCLUSIONS: This data may help to explain the anti-carcinogenic and cardioprotective effects of tea catechins. The effect of tea catechins on Na(+)/K(+)-ATPase and NHE may be explained due to a direct effect of these compounds on plasma membrane leading to a change in membrane fluidity. PMID- 15748599 TI - Relationship between plasma lipid concentrations and HDL subclasses. AB - BACKGROUND: It is generally accepted that different high-density lipoprotein (HDL) subclasses have distinct but interrelated metabolic functions. HDL is known to directly influence the atherogenic process and changes in HDL subclasses distribution may be related to the incidence and prevalence of atherosclerosis. METHODS: Apo-AI contents(mg/l) of plasma HDL subclasses were determined by 2 dimensional gel electrophoresis coupled with immunodetection for apo-AI. Four hundred forty-two Chinese adults subjects aged 33 to 78 years were assigned to different groups according to the third Report of NCEP (ATP III) guidelines. The subjects were first divided into 2 groups, normal and high TG, then further classified by plasma TC, HDL-C and LDL-C concentrations. The subjects were also divided into TC desirable and TC high groups. RESULTS: Apo-A contents of prebeta(1)-HDL were higher while HDL(2b) were lower in high TG subjects vs. the corresponding normal TG subjects according to plasma TC and LDL-C concentrations. With the increase of plasma TC concentrations, apo-AI contents of prebeta(1)-HDL were significantly higher in high TC subgroup vs. TC desirable subgroup in normal TG subjects. With the decrease of HDL-C concentrations, apo-AI contents of HDL(2b) tended to decrease in normal TG subjects. And, with the increases of LDL C concentration, in normal TG subjects, apo-AI contents of prebeta(1)-HDL and HDL(3b) were significantly higher and those of HDL(2b) were significantly lower in very high LDL-C subgroup vs. LDL-C optimal subgroup. On the other hand, apo-AI contents of prebeta(1)-HDL and HDL(3a) were significantly higher, while HDL(2a) and HDL(2b) were significantly lower in high TG and very high TG subgroup vs. normal TG subgroup within either TC desirable or TC high subjects. In a multivariate linear regression model, TG and TC concentrations were all associated independently and positively with high prebeta(1)-HDL; however, HDL-C were inversely associated with high prebeta(1)-HDL. And TG and TC concentrations were all associated independently and negatively with low HDL(2b), but HDL-C and apo-AI were positively associated with low HDL(2b). CONCLUSIONS: With the increase of plasma TG, TC, LDL-C or the decrease of plasma HDL-C concentrations, there was a general shift toward smaller-sized HDL, which, in turn, indicates that reverse cholesterol transport might be weakened and HDL maturation might be abnormal. Plasma TG concentration is a more important factor than TC concentration on the changes of HDL subclass distribution. Moreover, when TG is normal and HDL-C decreased, large-size HDL particles tended to decrease. PMID- 15748602 TI - Serological diagnosis of infectious mononucleosis by chemiluminescent immunoassay using capsid antigen p18 of Epstein-Barr virus. AB - BACKGROUND: Infectious mononucleosis is the common clinical manifestation of primary Epstein-Barr virus (EBV) infection in young children. We evaluated a chemiluminescent immunoassay method for the determination of serum anti-viral capsid antigen IgM antibody and its clinical value in the diagnosis of infectious mononucleosis. METHODS: Concentrations of the antibody in serum samples from 187 children measured by chemiluminescent immunoassay were compared with those measured by ELISA. RESULTS: Assessment of technologic quality (methodology) in diagnostic tests demonstrated that sensitivity of CLIA was 0.64 U/ml and the functional sensitivity was <0.9 U/ml. The within-assay and the between-assay imprecisions of different concentrations were all <5%. Recoveries were all in 93 107%. The linear regression equation between expected values and measured values was y=0.0967+1.0093x, correlation coefficient was 0.9996 (p<0.0001). The ROC curve showed that the sensitivity and specificity of the CLIA both were >90%. The area under the curve was 0.992, which was significantly higher than that of ELISA (p<0.05). CONCLUSION: The CLIA was the excellent method for EBV-VCA IgM measurement at present and can improve the clinical diagnosis of infectious mononucleosis. PMID- 15748601 TI - Relationship between lipocalin-type prostaglandin D synthase and alpha glucosidase in azoospermia seminal plasma. AB - BACKGROUND: [corrected] To determine the correlation of lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase in semen. METHODS: We analyzed 68 seminal plasmas for lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase, L-PGDS was analyzed by ELISA. The semen donors were categorized in 3 groups: normal, obstructive and non-obstructive azoospermia. We then evaluated their correlation. RESULTS: The difference of L-PGDS concentration (P<0.001) and alpha-glucosidase activity (P<0.001) among the 3 clinical groups was statistically significant. Correlation between L-PGDS concentration and alpha glucosidase was also statistically significant. L-PGDS concentration correlated positively with alpha-glucosidase activity (r=0.882). CONCLUSIONS: L-PGDS in seminal plasma, like alpha-glucosidase, suggests an obstruction of the seminal ducts and may be a potential marker that may aid in the differential diagnosis of obstructive and non-obstructive azoospermia. PMID- 15748603 TI - Analysis of plasma amino acids by HPLC with photodiode array and fluorescence detection. AB - BACKGROUND: Plasma amino acids are usually analyzed by ion-exchange chromatography (IEC), a reproducible but time consuming method. Here, we test whether plasma amino acids can be analyzed using reverse-phase high performance liquid chromatography (HPLC). METHODS: Filtered plasma, with S-carboxymethyl-l cysteine as the internal standard, was derivatized and analyzed by an Agilent 1100 HPLC system. Primary amino acids were derivatized with o-phthalaldehyde 3 mercaptopropionic acid (OPA) and detected by a diode array detector. Secondary amino acids were derivatized with 9-fluorenylmethyl chloroformate (FMOC) and detected fluorometrically. Chromatographic separation is achieved by two gradient elutions (two injections per sample), starting at different pHs, on a reverse phase Agilent Zorbax Eclipse C(18) column AAA (4.6 x 150 mm). RESULTS: The HPLC method evaluated correlated well with IEC (0.897 g/l and intralipid >5 g/l. No interference was observed from bilirubin or ascorbic acid. The assay was in good agreement with values obtained by HPLC and fluorescence polarization immunoassay. CONCLUSIONS: The performance of the Catch homocysteine assay was acceptable on the Hitachi 917 chemistry analyzer. PMID- 15748608 TI - Anti-cyclic citrullinated peptide autoantibodies in IgM rheumatoid factor positive patients. AB - BACKGROUND: Antibodies to citrullinated proteins have been described in patients with RA and these appear to be the most specific markers of the disease. The objective of this study was to analyse the improvement in diagnostic accuracy of anti-cyclic citrullinated peptide autoantibodies and IgA rheumatoid factor in patients with clinical suspicion of RA and who were IgM rheumatoid factor positive. Anti-CCP antibodies were measured with three different second generation enzyme immunoassays. METHODS: We chose 133 serum samples with IgM RF levels greater than 20 IU/mL sent to our Laboratory from Specialized Care Units. Subsequently, patients were classified according to their clinical records. Eighty-seven had rheumatoid arthritis and 46 had other diseases. In all samples anti-CCP and IgA RF were measured by the corresponding ELISAs. RESULTS: Comparison of the three anti-CCP second-generation ELISAs revealed differences between them. Likewise, clinical performances in terms of sensitivity, specificity, and positive and negative likelihood ratios were different. In patients with IgM RF higher than 20 IU/mL, anti-CCP antibodies increased the clinical efficiency of IgM RF and offered better performance as compared with IgA RF. CONCLUSIONS: The use of anti-CCP antibodies affords good clinical efficiency and modifies the pre-test probability of the occurrence of RA in patients with IgM rheumatoid factor higher than 20 IU/mL. PMID- 15748609 TI - G(M1)-ganglioside degradation and biosynthesis in human and murine G(M1) gangliosidosis. AB - BACKGROUND: Gangliosides are building blocks of cell membranes and their biosynthesis and degradation have been extensively studied in the past. Regulation of the metabolism of these glycolipids controls fundamental cell functions. G(M1)-gangliosidosis, a neurodegenerative glycosphingolipid storage disease, is caused by deficiency of lysosomal beta-galactosidase with consequent disruption of the normal degradative pathway of G(M1)-ganglioside. We studied the impact of G(M1)-ganglioside accumulation on its biosynthetic enzyme in cells and tissues from human patients and from the G(M1)-gangliosidosis mouse model. METHODS: We tested the qualitative and quantitative pattern of gangliosides by thin layer chromatography and N-acetylneuraminic acid dosage, respectively. Regulation of G(M1)-ganglioside biosynthesis was evaluated by G(M1) synthase assay in human and murine samples. RESULTS: G(M1)-ganglioside accumulation has an inhibitory effect on the human but not on the mouse G(M1) synthase. We present evidence that G(M1) synthase activity in human and murine cells are regulated by different mechanisms. CONCLUSIONS: Alternative pathways in the mouse may account for these results and possibly explain some of the phenotypical differences between the human and mouse forms of this disorder. PMID- 15748610 TI - Age-related changes in the vasodilating actions of Ginkgo biloba extract and its main constituent, bilobalide, in rat aorta. AB - BACKGROUND: Age-related modulation in vasodilating actions induced by Ginkgo biloba extract (GBE) and bilobalide, a main constituent of GBE, were examined using rat aorta ring strips. METHODS: Wistar rats from 5 to 25 weeks old were used, and the isolated aorta ring strips were fixed in Krebs-Henseleit solution. RESULTS: GBE and bilobalide concentration-dependently dilated norepinephrine (NE) induced vasoconstriction in all aged rats. The vasodilating actions generally decreased in accordance with aging. GBE at 1 mg/ml decreased from 28.4+/-3.8% (n=5) in 5-week-old rats to 23.7+/-7.1 (n=7) in 25-week-old rats, but not significantly. GBE (3 mg/ml)-induced vasodilation was maximum by 73.7+/-2.1% (n=4, P<0.001) in 10-week-old rats. GBE had the marked vasodilation at younger ages and further decreased it with developing ages. In the rats older than 20 weeks, however, GBE tended to rather increase the strength of vasodilating action. On the other hand, the vasorelaxation induced by 30 micromol/l bilobalide significantly decreased from 11.8+/-1.4% (n=4) in 5-week-old rats to 2.3+/-1.5% (n=5, P<0.01) in 25-week-old rats, and by 100 micromol/l from 20.2+/-3.4% (n=4) to 5.6+/-2.5% (n=5, P<0.01), respectively. Bilobalide had the similar age-related actions. The age-dependent attenuation was produced milder by bilobalide than by GBE. At lower concentrations, however, bilobalide caused the weak vasocontriction in 20- and 25-week-old rats. CONCLUSION: GBE and bilobalide possess a similar characteristic for age-related modification, clinically suggesting the more effective actions of GBE for elder persons. PMID- 15748611 TI - Quantitative determination of vigabatrin and gabapentin in human serum by gas chromatography-mass spectrometry. AB - BACKGROUND: Published methods for routine clinical monitoring of vigabatrin and gabapentin are often very laborious. A simple GC-MS method was developed for the simultaneous quantitative determination of vigabatrin and gabapentin in human serum. METHODS: After protein precipitation, the compounds are derivatized by methylation and analysed on a polydimethylsiloxane column using splitless injection. Cyclobarbital is used as the internal standard. To attain maximal sensitivity, detection is performed in selected ion monitoring mode. RESULTS: The method was fully validated and linear calibration curves were obtained in the concentration ranges from 5 to 80 microg/mL for vigabatrin and from 5 to 30 microg/mL for gabapentin. The within-day and day-to-day relative standard deviations at three different concentration levels were <10% and <15%, respectively. The limit of quantitation was 2 mug/mL for both compounds. CONCLUSIONS: The presented method provides high chromatographic resolution, good sensitivity and unequivocal identification potential and can be used for simultaneous analysis of both antiepileptics. PMID- 15748612 TI - eNOS gene intron 4 a/b VNTR polymorphism is a risk factor for coronary artery disease in Southern Turkey. AB - BACKGROUND: Nitric oxide (NO) plays a major role in the regulation of vascular tone Associations between NO genotypes, coronary artery disease (CAD) and other risk factors have been described by many authors. The aim of this study was to investigate the role of endothelial nitric oxide synthase (eNOS) gene intron 4 a/b variable number of tandem repeats (VNTR) polymorphism and other risk factors in the development of CAD in subjects living in Southern Turkey. METHODS: Two hundred and sixty-six patients (154 males and 112 females, aged between 30 and 80 years, mean 52.4+/-10.3) whose coronary arteries were evaluated by means of coronary angiography were enrolled in the study. Of the total, 133 had CAD (Group I) and the remaining had normal coronary arteries (Group II). The eNOS gene intron 4 a/b VNTR polymorphism was analyzed by polymerase chain reaction. The plasma lipid levels and other risk factors were also determined in all subjects. RESULTS: The a allele frequencies and genotypes carrying a allele were significantly higher in Group I. Plasma lipids, except HDL-C, were also increased in this group. We found that hypertension (HT), diabetes mellitus (DM), male gender, age and smoking were the independent predictors of CAD. CONCLUSION: a allele of eNOS intron 4 a/b VNTR polymorphism is not an independent predictor of CAD. eNOS intron 4 a/b polymorphism (presence of a allele) is a risk factor in addition to HT, DM, male gender, age and smoking for the development of CAD in Southern Turkey. PMID- 15748614 TI - Screening of newborns and high-risk group of children for inborn metabolic disorders using tandem mass spectrometry in South Korea: a three-year report. AB - BACKGROUND: Mass screening using tandem mass spectrometry(MS/MS) was initiated to determine if the incidence of metabolic disorder is sufficiently high to meet the criteria for newborn screening, and whether or not early medical intervention might be beneficial to the patients. METHODS: Newborns and children in a high risk group were screened using MS/MS from April 2001 to March 2004. Blood spots of newborns were collected between 48 and 72 h after birth. The dried blood spots was extracted with 150 microl of methanol, and analyzed by MS/MS. RESULTS: From April 2001 to March 2004, 79,179 newborns were screened for organic, amino and fatty acid metabolism disorders, which account for approximately 5.4% of annual births in South Korea. Twenty-eight newborns were diagnosed with one of the metabolic disorders and the collective estimated prevalence amounted to 1 in 2800 with a sensitivity of 97.67%, a specificity of 99.28%, a recall rate of 0.05%, and a positive predictive value of 6.38%. 6795 infants/children at high risk were screened and 20 were confirmed to have metabolic disorders. CONCLUSION: The collective total prevalence of 1:2800 in newborns indicates an underestimation of the incidence of metabolic disorders prior to implementing MS/MS screening in South Korea. PMID- 15748613 TI - Counteraction of oxalate induced nitrosative stress by supplementation of l arginine, a potent antilithic agent. AB - BACKGROUND: Our understanding of nitrosative stress in the process of urolithiasis is far from complete. Earlier studies carried out in our laboratory demonstrate the presence of nitrated THP in stone formers, l-arginine (l-arg) a precursor of nitric oxide (NO), attenuates the endothelial dysfunction caused by reactive nitrogen species. We investigated the role of l-arg in ethylene glycol (EG)-induced urolithic rat model and observed its antilithic and antioxidative properties. METHODS: Hyperoxaluria was induced using 0.75% EG in drinking water. l-arg [1.25 g/kg body weight] was given orally for a period of 28 days. RESULTS: EG-treated rats showed significant loss in body weight and increase in the activities of oxalate synthesizing enzymes such as glycollic acid oxidase in liver. Lactate dehydrogenase activity in liver and kidney was increased. The activity of the free radical producing enzyme xanthine oxidase, tissue oxalate and calcium levels were significantly increased in EG-treated rats. Depletion in the antioxidant enzymes, membrane bound ATPases and thiol status was observed in these rats. l-arg co-supplementation to EG-treated rats maintained the activities of the oxalate synthesizing enzymes and free radical producing enzymes with in the normal range. Tissue oxalate and calcium levels were also maintained near normal in l-arg treated hyperoxaluric rats. l-arg, by its cytoprotective effect, maintained the thiol status, thereby preserving the activities of the membrane bound ATPases and preventing proteinuria and subsequent weight loss in EG-treated rats. CONCLUSION: l-arg feeding prevents the retention of calcium oxalate crystals in hyperoxaluric rats by way of protecting the renal cells from oxidative injury and also by providing a second line of defense through the normalization of the oxalate metabolism. It reduces the risk of stone formation, by curtailing free radicals and hyperoxaluria as both of them have to work in close association to form stones. PMID- 15748615 TI - Diagnosis accuracy of a new challenger for thrombosis exclusion, the Stratus CS DDMR. AB - BACKGROUND: The diagnostic accuracy of a new Point of Care, rapid and quantitative D-dimer assay (Stratus CS DDMR from Dade Behring) was evaluated. METHODS: Vidas test from bioMerieux was used as reference method in 279 patients recruited from a management study in progress in our institution. RESULTS: Both assays show comparable reproducibility (2.9% with the Stratus CS DDMR and 4.4% with the D-dimer Vidas in the cut-off range) and good correlation (R(2)=0.9057). The overall test performance as assessed by the area under the curve of the ROC curves is 0.801 for the Stratus CS DDMR assay and 0.798 for the D-dimer Vidas assay. By using assay regression curves, likelihood ratios or test agreement approaches, a 440-450 ng/ml value is evidenced as the threshold value for the Stratus CS DDMR, which nears at best the performances of the D-dimer Vidas 500 or 550 ng/ml threshold value. This proposed exclusion value ensures a 95% sensitivity and a 45% specificity. The few false negative results using the two assays only evidenced sub-popliteal thromboses, that would not have been considered as having thrombosis if an above-the-knee test had been performed. In these conditions, sensitivity would have been 100%. CONCLUSIONS: A new quantitative D-dimer assay, the Stratus CS DDMR, demonstrated performances comparable with those of the DD Vidas test. PMID- 15748616 TI - Detection and allele-frequencies of the 833T>C, 844ins68 and a novel mutation in the cystathionine beta-synthase gene. AB - BACKGROUND: The most common 833T>C/844ins68 in cis double mutation in the cystathionine beta synthase (CBS) gene probably is non-pathogenic because the 68 bp insertion eliminates the 833T>C mutation due to alternative splicing. However, allele frequency and effects of the isolated 833T>C mutation are unclear. METHOD: DNA was isolated from 500 volunteers and used directly for PCR-RFLP of CBS gene exon-8. A new primer design was developed to create annealing sites upstream and downstream of exon-8 for forward and reverse primers, respectively. The design was made to exclude sequence homology of the forward primer with the insert fragment and to introduce an internal Bsr1 digestion site. RESULTS: A new 9276G>A mutation was found in intron 8. Because of this mutation, an extra Bsr1 digestion site is created in intron 8. In Caucasian volunteers, the following allele frequencies were found: 833T>C=0.2%, 833T>C/844ins68=10.2%, and 9276G>A=0.2%. CONCLUSION: The developed PCR-RFLP method is able to detect the 833T>C mutation, the 833T>C/844ins68 polymorphism as well as a new 9276G>A mutation in intron 8. Further study should explore the effect of the isolated 9276G>A mutation. PMID- 15748617 TI - Long-chain omega6 polyunsaturated fatty acids in erythrocyte phospholipids are associated with insulin resistance in non-obese type 2 diabetics. AB - BACKGROUND: Associations have been described between insulin action and long chain omega6 polyunsaturated fatty acids in skeletal muscle and erythrocytes of healthy individuals. We evaluated whether there was an association between insulin resistance and long-chain omega6 polyunsaturated fatty acids in erythrocyte phospholipids of type 2 diabetics. METHODS: Erythrocyte phospholipid fatty acid composition was determined in 43 adult non-obese type 2 diabetics. Insulin resistance was estimated by the HOMA mathematical approximations. Individual omega6 fatty acids and the sum of long-chain omega6 polyunsaturated fatty acids were correlated with insulin resistance. The same association was assessed for fatty acid product/precursor ratios taken as measures of Delta5 desaturase, Delta6 desaturase and their combined action. RESULTS: Except for 18:2omega6, 20:2omega6 and 20:3omega6, the individual omega6 fatty acids tested and the sum of long-chain omega6 polyunsaturated fatty acids correlated negatively with insulin resistance at p<0.005. Likewise, a negative correlation was found for the combined action of Delta5 and Delta6 desaturases (p<0.005). CONCLUSIONS: Our results indicate that formation of long-chain omega6 fatty acids from 18:2omega6 by desaturation-elongation is impaired under insulin resistant conditions and reduced desaturase activities may be involved. In view of our results, we hypothesize that in insulin resistant conditions, insulin-mediated activation of Delta5 and Delta6 desaturases may not occur. PMID- 15748618 TI - Major discrepancy between digoxin immunoassay results in a context of acute overdose: a case report. PMID- 15748619 TI - Effects of simvastatin on exercise-induced myocardial ischemia and plasma endothelin-1 concentrations in patients with stable angina. PMID- 15748620 TI - Prognostic value of postoperative cTnI in cardiac surgery: comparison of Abbott and Dade assays. PMID- 15748621 TI - Prevalence of increased Tn I measurements using different assays. PMID- 15748622 TI - Characterization of chromanol 293B-induced block of the delayed-rectifier K+ current in heart-derived H9c2 cells. AB - The effects of chromanol 293B on ion currents in rat embryonic heart-derived H9c2 cells were investigated in this study. Chromanol 293B suppressed the amplitude of delayed rectified K+ current (I(K)) in a concentration-dependent manner. The IC50 value for chromanol 293B-induced inhibition of I(K) was 8 microM. The I(K) present in these cells, the electrical properties of which resembled those for the Kv2.1-related K+ current, was sensitive to inhibition by quinidine or dendrotoxin, yet not by pandinotoxin-Kalpha, E-4031 or apamin. Chromanol 293B reduced the activation time constant of I(K) and the effective gating charge of this channel. However, little or no modification in the steady-state inactivation of I(K) in response to long-lasting conditioning pulses could be demonstrated in the presence of chromanol 293B. These results clearly demonstrate that chromanol 293B can effectively interact with the K+ channel functionally expressed in H9c2 myoblasts. The chromanol 293B-induced inhibition of these channels could primarily be attributed to open channel block. PMID- 15748623 TI - Effect of thiopental, pentobarbital and diethyl ether on early steps of insulin action in liver and muscle of the intact rat. AB - A large number of experimental studies have investigated insulin signaling in rats. In these studies different anaesthetics have been used to anaesthetize rats. However, the direct effects of anaesthetics on the regulation of the early steps of insulin action are not known. In the present study, we investigated the effect of thiopental, pentobarbital and diethyl ether on the plasma glucose disappearance rate, IR, IRS-1 and IRS-2 tyrosine phosphorylation, IRSs association with PI 3-kinase, Akt and Erk phosphorylation, in liver and muscle of rats. Fasting plasma glucose levels were higher in animals anaesthetized with ether. No differences in plasma glucose disappearance rates were observed, however. Insulin-induced IR, IRS-1 and IRS-2 tyrosine phosphorylation, association of these substrates with PI 3-kinase and Akt and ERK phosphorylation were similar in the three groups of animals in both tissues. These data suggest that both thiopental and pentobarbital may be used in studies where changes in insulin signaling are being measured and where adequate general anaesthesia is required. PMID- 15748624 TI - Toxicogenomics of resveratrol in rat liver. AB - Resveratrol, a polyphenolic compound found in grape skin and peanuts has been shown to prevent many diseases including cardiovascular diseases and cancer. To better understand resveratrol's potential in vivo toxicity, we studied the dose response using cDNA stress arrays coupled with drug metabolizing enzymatic (DME) assays to investigate the expression of stress-responsive genes and Phase I and II detoxifying enzymes in rat livers. Male and female CD rats were treated with high doses of resveratrol (0.3, 1.0 and 3.0 gm/kg/day) for a period of 28 days. Total RNA from rat liver was reverse-transcribed using gene-specific primers and hybridized to stress-related cDNA arrays. Among female rats, Phase I DME genes were repressed at 0.3 and 1.0 gm/kg/day doses, while genes such as manganese superoxide dismutase, cytochrome P450 reductase, quinone oxidoreductase and thiosulfate sulfurtransferase demonstrated a dose-dependent increase in gene expression. The modulation of these liver genes may implicate the potential toxicity as observed among the rats at the highest dose level of resveratrol. Real-Time PCR was conducted on some of the Phase II DME genes and anti-oxidant genes to validate the cDNA array data. The gene expression from real-time PCR demonstrated good correlation with the cDNA array data. UGT1A genes were amongst the most robustly induced especially at the high doses of resveratrol. We next performed Phase I and Phase II enzymatic assays on cytochrome P450 2E1 (CYP2E1), cytochrome P450 1A1 (CYP1A1), NAD(P)H:quinone oxidoreductase (NQO1), glutathione S-transferase (GST) and UDP-glucuronosyl transferase (UGT). Induction of Phase II detoxifying enzymes was most pronounced at the highest dose of resveratrol. CYP1A1 activity demonstrated a decreasing trend among the 3 dose groups and CYP2E1 activity increased marginally among female rats over controls. In summary, at lower doses of resveratrol there are few significant changes in gene expression whereas the modulation of liver genes at the high dose of resveratrol may implicate the potential toxicity observed. PMID- 15748626 TI - Local effect of vanadate on interstitial glucose and lactate concentrations in human skeletal muscle. AB - The aim of this study was to investigate the local effect of the insulin-mimetic agent vanadate on glucose metabolism in human skeletal muscle in vivo. Interstitial concentrations of glucose and lactate were determined by microdialysis at a low flow rate in the quadriceps femoris muscle of 18 men. In the same leg two microdialysis catheters were inserted. In one catheter, the perfusion medium was supplemented with sodium metavanadate (10-100 mM) after a basal period, the other catheter served as control. In the catheter perfused with metavanadate, the interstitial glucose concentration was decreased by 13-50% compared to the control catheter (p<0.05). The lactate concentration was higher in the 50 mM and 100 mM metavanadate catheters compared to control (39-89%, p<0.05). There was no difference between control and metavanadate catheters in urea concentrations. Five of the subjects were insulin-resistant and for them the results were similar, although the effect was somewhat smaller. The decreased interstitial glucose concentration, and the increased lactate concentration, in the vicinity of the microdialysis catheter most likely reflects an increased cellular glucose uptake. The present study thus indicates that vanadate mimics the effect of insulin in human skeletal muscle in vivo. PMID- 15748625 TI - Inhibition of inducible nitric oxide synthase and cyclooxygenase II by Platycodon grandiflorum saponins via suppression of nuclear factor-kappaB activation in RAW 264.7 cells. AB - Saponins are glycosidic compounds present in many edible and inedible plants. They exhibit potent biological activities in mammalian systems, including several beneficial effects such as anti-inflammation and immunomodulation. In this study, we investigated the effects of seven platycodin saponins on the activities of inducible nitric oxide synthase (iNOS) and cyclooxygenase II (COX-2) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. We found that 2"-O-acetyl polygalacin D (S1), platycodin A (S2), platycodin D (S3), and polygalacin D (S6) inhibited LPS-induced NO production in a concentration-dependent manner. Furthermore, these compounds inhibited the expression of LPS-induced iNOS and COX 2 protein and mRNA without an appreciable cytotoxic effect on RAW 264.7 macrophages, and could suppress induction by LPS of pro-inflammatory cytokines such as prostaglandin E2 (PGE2). Treatment with these compounds of RAW 264.7 cells transfected with a reporter construct indicated a reduced level of LPS induced nuclear factor-kappaB (NF-kappaB) activity and effectively lowered NF kappaB binding as measured by electrophoretic mobility shift assay (EMSA). The suppression of NF-kappaB activation appears to occur through the prevention of inhibitor kappaB (IkappaB) degradation. In vivo, platycodin saponin mixture (PS) and S3 protected mice from the lethal effects of LPS. The 89% lethality induced by LPS/galactosamine was reduced to 60% and 50% when PS and S3, respectively, were administered simultaneously with LPS. These results suggest that the main inhibitory mechanism of the platycodin saponins may be the reduction of iNOS and COX-2 gene expression through blocking of NF-kappaB activation. PMID- 15748627 TI - Effect of excitotoxic lesions of the neonatal ventral hippocampus on the immobility response in rats. AB - Rats with neonatal ventral hippocampal (nVH) lesions show postpubertal hypersensitivity to dopamine agonists, which may be reversed by neuroleptic treatment. In addition, the immobility response (IR) may be regulated by dopaminergic activity. We investigated the influence of the IR caused by clamping the neck of rats that had received bilateral ibotenic acid lesions of the ventral hippocampus at postnatal day 7 (PD7). At both ages, prepubertal (PD35) and postpubertal (PD56), the duration of the IR was significantly increased in animals with lesions when compared to controls. These findings indicate that nVH damage results in behavioral changes, such as enhancement of the IR, related to mesolimbic dopaminergic transmission. PMID- 15748628 TI - Evidence for a spinal serotonergic control of the peripheral inflammation in the rat. AB - We investigated the effect of serotonergic agonists and antagonists injected intrathecally by direct punction of the spinal cord at the lumbar level (between L5-L6) on peripheral inflammatory edema. Edema was induced by carrageenan injected subcutaneously in one hindpaw 30 min after spinal treatments. Serotonin (0.1, 1, 10 pmol) caused a graded-inhibition of the inflammatory paw edema. The corticosteroid inhibitor aminoglutethimide (100 mg/kg, p.o. 1.5 h before spinal treatment) did not modify this effect. The 5-HT1A agonist buspirone and the 5 HT1B/1D agonist sumatriptan (0.1, 1.0 and 10 nmol) also inhibited paw edema. The 5-HT1,2 antagonist methysergide (10 and 100 pmol) enhanced edema, but higher doses ( 4 and 8 nmol) diminished edema. NAN-190 (5-HT1 antagonist; 1 and 10 nmol) increased paw edema, while ritanserin (5-HT2 antagonist; 1 nmol) inhibited paw edema. Ondansetron (5-HT3 antagonist; up to 10 nmol) did not affect edema, but metoclopramide (5-HT3 antagonist / 5-HT4 agonist; 5, 10 and 30 pmol) inhibited edema. These data suggest that a tonic release of serotonin in the spinal cord may occurs during ongoing peripheral inflammation, modulating the neurogenic component of edema either by an inhibitory action on 5-HT1 receptors or by a stimulatory action on 5-HT2 receptors. A disfunction in such mechanism may be involved in the pathophysiology of certain types of headaches or migraine, which seem to depend on neurogenic vasodilation, and may also help to explain the therapeuthic effectiveness of some serotonergic agents in these conditions. PMID- 15748629 TI - Cerebral salt wasting in subarachnoid hemorrhage rats: model, mechanism, and tool. AB - Cerebral salt wasting (CSW) frequently occurs concomitantly with aneurysmal subarachnoid hemorrhage (SAH). CSW induces excessive natriuresis and osmotic diuresis, and reduces total blood volume. As a result, the risk of symptomatic cerebral vasospasm may be elevated. Therefore, it is important to determine the mechanism of CSW. The purpose of this study was to evaluate whether the rat SAH model exhibits CSW and to investigate the relationship between CSW and natriuretic peptides. A SAH model was produced in 24 rats by perforating a cerebral artery with a nylon thread up through the common carotid artery. To evaluate CSW, urine was cumulatively collected from SAH onset to 12 hours and sodium (Na) excretion was analyzed. Body weight and hematocrit were analyzed before and after SAH onset. Concentrations of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in plasma were also analyzed. Urine volume and total Na excretion of SAH rats were significantly higher than those of sham rats (p<0.05). Body weight of SAH rats significantly decreased and hematocrit significantly increased (p < 0.05). ANP concentration was significantly decreased in SAH rats (p<0.05). However, BNP concentrations did not change. This study demonstrated for the first time that a rat SAH model exhibited CSW. It was suggested that the cause of CSW was neither ANP nor BNP. In addition, this rat SAH model will be useful for study of CSW after SAH. PMID- 15748630 TI - Physiological stress response to video-game playing: the contribution of built-in music. AB - Recent studies on video game playing have uncovered a wide range of measurable physiological effects on the organism, such as increases in cardiovascular activity and breathing responses. However, the exact source of these effects remains unclear. Given the well-known effects of sound on physiological activity, especially those of noise and of music, and on the secretion of the stress hormone cortisol in particular, we hypothesized that music may be a major source of stress during video game playing. We thus examined the effect of built-in music on cortisol secretion as a consequence of video game playing. Players were assigned quasi-randomly to either a Music or a Silence condition. Four saliva samples were taken, that is, after practice (T1), immediately after having played for 10 minutes (T2), 15 minutes after the end of the experiment (T3), and 30 minutes after the end of the experiment (T4). The results show that the Music group had significantly higher cortisol levels at T3, that is, when cortisol levels are assumed to reflect the stress induced by the game. These findings suggest for the first time that the auditory input contributes significantly to the stress response found during video game playing. PMID- 15748631 TI - Metabolite-cytochrome P450 complex formation by methylenedioxyphenyl lignans of Piper cubeba: mechanism-based inhibition. AB - Five methylenedioxyphenyl lignans, (-)-clusin (1), (-)-dihydroclusin (2), (-) yatein (3), (-)-hinokinin (4), and (-)-dihydrocubebin (5), were isolated from Piper cubeba as potent and selective inhibitors against cytochrome P450 3A4 (CYP3A4). In this study, we investigated the mechanism of inhibition of CYP3A4 by these lignans and the possibility of their mechanism-based inhibition. Using [N methyl-14C]erythromycin as a substrate, all lignans appear to be showed mixed type of inhibition with apparent Ki of 1.96-4.07 microM. Furthermore, all lignans (1-5) inhibited CYP3A4 in a time-, concentration-, and NADPH-dependent manners and thus appear to be the mechanism-based inhibitors of CYP3A4. The apparent inactivation parameter, K(I) for these compounds were in the range of 0.054-0.373 microM, whereas the k(inact) values were 0.225-0.320 min-1. Among them, (-) clusin (1) and (-)-dihydroclusin (2) were found to be the most potent CYP3A4 inactivator with apparent K(I) and k(inact) values of 0.082, 0.054 microM and 0.253, 0.310 min-1, respectively. Spectral scanning of microsomes with these lignans yielded an absorbance at 455 nm, suggesting that all of them appear to inactivate the cytochrome P450 via the formation of a metabolite intermediate complex. This pattern is consistent with the metabolism of the methylenedioxyphenyl compounds. These results indicate that (-)-clusin (1), (-) dihydroclusin (2), (-)-yatein (3), (-)-hinokinin (4), and (-)-dihydrocubebin (5) are effective mechanism-based inhibitors of CYP3A4. PMID- 15748632 TI - Lack of effect of ghrelin treatment on melatonin production in rat pineal and Harderian glands. AB - The effects of ghrelin, a peptide hormone secreted from the stomach, on melatonin remain unknown. The aim of the study was to investigate possible ghrelin melatonin interactions by studying the effect of ghrelin treatment on melatonin production in rat pineal and Harderian glands. Young (9 weeks) and old (20 months) male Wistar rats, maintained under a light:dark cycle regimen of 12:12, were assigned randomly to either a single subcutaneous (s.c.) injection of saline or ghrelin (1 microg/rat or 15 microg/rat) 1 h before sacrifice in the middle of the dark phase, or repeated s.c. saline or ghrelin injections (15 microg/rat), 3, 2 and 1 h before sacrificed in the middle of the dark phase. Neither ghrelin doses (1 microg/rat or 15 microg/rat) nor type of treatment (acute or repeated) influenced melatonin levels or the melatonin synthesizing enzymes N acetyltransferase and hydroxyindole-O-methyltransferase activities, either in pineal gland or in Harderian glands. At the concentrations used, ghrelin does not influence melatonin production in rat pineal and Harderian glands, and therefore is not involved in the regulation of melatonin secretion, at least under our experimental conditions. PMID- 15748634 TI - Ultraviolet radiation-mediated damage to cellular DNA. AB - Emphasis is placed in this review article on recent aspects of the photochemistry of cellular DNA in which both the UVB and UVA components of solar radiation are implicated individually or synergistically. Interestingly, further mechanistic insights into the UV-induced formation of DNA photoproducts were gained from the application of new accurate and sensitive chromatographic and enzymic assays aimed at measuring base damage. Thus, each of the twelve possible dimeric photoproducts that are produced at the four main bipyrimidine sites can now be singled out as dinucleoside monophosphates that are enzymatically released from UV-irradiated DNA. This was achieved using a recently developed high-performance liquid chromatography-tandem mass spectrometry assay (HPLC-MS/MS) assay after DNA extraction and appropriate enzymic digestion. Interestingly, a similar photoproduct distribution pattern is observed in both isolated and cellular DNA upon exposure to low doses of either UVC or UVB radiation. This applies more specifically to the DNA of rodent and human cells, the cis-syn cyclobutadithymine being predominant over the two other main photolesions, namely thymine-cytosine pyrimidine (6-4) pyrimidone adduct and the related cyclobutyl dimer. UVA irradiation was found to generate cyclobutane dimers at TT and to a lower extent at TC sites as a likely result of energy transfer mechanism involving still unknown photoexcited chromophore(s). Oxidative damage to DNA is also induced although less efficiently by UVA-mediated photosensitization processes that mostly involved 1O2 together with a smaller contribution of hydroxyl radical mediated reactions through initially generated superoxide radicals. PMID- 15748635 TI - Mutations induced by ultraviolet light. AB - The different ultraviolet (UV) wavelength components, UVA (320-400 nm), UVB (280 320 nm), and UVC (200-280 nm), have distinct mutagenic properties. A hallmark of UVC and UVB mutagenesis is the high frequency of transition mutations at dipyrimidine sequences containing cytosine. In human skin cancers, about 35% of all mutations in the p53 gene are transitions at dipyrimidines within the sequence 5'-TCG and 5'-CCG, and these are localized at several mutational hotspots. Since 5'-CG sequences are methylated along the p53 coding sequence in human cells, these mutations may be derived from sunlight-induced pyrimidine dimers forming at sequences that contain 5-methylcytosine. Cyclobutane pyrimidine dimers (CPDs) form preferentially at dipyrimidines containing 5-methylcytosine when cells are irradiated with UVB or sunlight. In order to define the contribution of 5-methylcytosine to sunlight-induced mutations, the lacI and cII transgenes in mouse fibroblasts were used as mutational targets. After 254 nm UVC irradiation, only 6-9% of the base substitutions were at dipyrimidines containing 5-methylcytosine. However, 24-32% of the solar light-induced mutations were at dipyrimidines that contain 5-methylcytosine and most of these mutations were transitions. Thus, CPDs forming preferentially at dipyrimidines with 5 methylcytosine are responsible for a considerable fraction of the mutations induced by sunlight in mammalian cells. Using mouse cell lines harboring photoproduct-specific photolyases and mutational reporter genes, we showed that CPDs (rather than 6-4 photoproducts or other lesions) are responsible for the great majority of UVB-induced mutations. An important component of UVB mutagenesis is the deamination of cytosine and 5-methylcytosine within CPDs. The mutational specificity of long-wave UVA (340-400 nm) is distinct from that of the shorter wavelength UV and is characterized mainly by G to T transversions presumably arising through mechanisms involving oxidized DNA bases. We also discuss the role of DNA damage-tolerant DNA polymerases in UV lesion bypass and mutagenesis. PMID- 15748636 TI - UVR-induced G-C to C-G transversions from oxidative DNA damage. AB - Many oxidizing agents induce G-C to T-A and G-C to C-G transversions, and the frequency largely depends on the oxidative conditions. Guanine is the most oxidizable base among natural bases. The typical oxidative lesion product 8 oxoguanine (8-oxoG) is responsible for G-C to T-A transversion but not for G-C to C-G transversion, and 8-oxoG is more readily oxidized than guanine because of its lowered ionization potential. Recently, imidazolone (Iz), guanidinohydantoin (Gh) and spiroiminodihydantoin (Sp) have been demonstrated as oxidative lesion products of guanine and 8-oxoG, which could be responsible for G-C to C-G transversions by forming specific base pair formations. PMID- 15748637 TI - The role of UV induced lesions in skin carcinogenesis: an overview of oncogene and tumor suppressor gene modifications in xeroderma pigmentosum skin tumors. AB - Xeroderma pigmentosum (XP), a rare hereditary syndrome, is characterized by a hypersensitivity to solar irradiation due to a defect in nucleotide excision repair resulting in a predisposition to squamous and basal cell carcinomas as well as malignant melanomas appearing at a very early age. The mutator phenotype of XP cells is evident by the higher levels of UV specific modifications found in key regulatory genes in XP skin tumors compared to those in the same tumor types from the normal population. Thus, XP provides a unique model for the study of unrepaired DNA lesions, mutations and skin carcinogenesis. The high level of ras oncogene activation, Ink4a-Arf and p53 tumor suppressor gene modifications as well as alterations of the different partners of the mitogenic sonic hedgehog signaling pathway (patched, smoothened and sonic hedgehog), characterized in XP skin tumors have clearly demonstrated the major role of the UV component of sunlight in the development of skin tumors. The majority of the mutations are C to T or tandem CC to TT UV signature transitions, occurring at bipyrimidine sequences, the specific targets of UV induced lesions. These characteristics are also found in the same genes modified in sporadic skin cancers but with lower frequencies confirming the validity of studying the XP model. The knowledge gained by studying XP tumors has given us a greater perception of the contribution of genetic predisposition to cancer as well as the consequences of the many alterations which modulate the activities of different genes affecting crucial pathways vital for maintaining cell homeostasis. PMID- 15748638 TI - After sun reversal of DNA damage: enhancing skin repair. AB - UV-induced DNA damage has been directly linked to skin cancer, and DNA repair is an important protection against this neoplasm. This is illustrated by the genetic disease xeroderma pigmentosum wherein a serious defect in DNA repair of cyclobutane pyrimidine dimers dramatically increases the rate of skin cancer. In other instances in which skin cancer rates are elevated, deficits in DNA repair may also be one of the causal factors. For example, solid organ transplant patients have elevated rates of skin cancer that are correlated with the dose and length of exposure to immunosuppressive drugs (predominantly cyclosporine A (CsA) and ascomycin (FK506)-related tacrolimus). We have found that treatment of cultured epidermal cells with CsA or ascomycin inhibits their removal of DNA damage by about 20% at 24 h. In a further example, people with a polymorphism in the DNA repair gene 8-oxo-guanine glycosylase (OGG1) have an increased risk of skin cancer. We have found that the cells with this variant polymorphism have an increased sensitivity of about 20% to a broad range of cytotoxic agents. The DNA deficits caused by immunosuppressive drugs or the OGG1 polymorphism can be overcome by the delivery of DNA repair enzymes in liposomes. The data suggests that deficits in DNA repair, even if they are not as severe as in the case of XP, may contribute to increased rates of cancer, and that topical therapy with DNA repair enzymes may be a promising avenue for after-sun protection. PMID- 15748639 TI - Genetic tumor archeology: microdissection and genetic heterogeneity in squamous and basal cell carcinoma. AB - Carcinogenesis is a multi-step series of somatic genetic events. The complexity of this multi-hit process makes it difficult to determine each single event and the definitive outcome of such events. To investigate the genetic alterations in cancer-related genes, sensitive and reliable detection methods are of major importance for generating relevant results. Another critical issue is the quality of starting material which largely affects the outcome of the analysis. Microdissection of cells defined under the microscope ensures a selection of representative material for subsequent genetic analysis. Skin cancer provides an advantageous model for studying the development of cancer. Detectable lesions occur early during tumor progression, facilitating molecular analysis of the cell populations from both preneoplastic and neoplastic lesions. Alterations of the p53 tumor suppressor gene are very common in non-melanoma skin cancer, and dysregulation of p53 pathways appear to be an early event in the tumor development. A high frequency of epidermal p53 clones has been detected in chronically sun-exposed skin. The abundance of clones containing p53 mutated keratinocytes adjacent to basal cell (BCC) and squamous cell carcinoma (SCC) suggests a role in human skin carcinogenesis. Studies using p53 mutations as a clonality marker have suggested a direct link between actinic keratosis, SCC in situ and invasive SCC. Microdissection-based studies have also shown that different parts of individual BCC tumors can share a common p53 mutation yet differ with respect to additional alterations within the p53 gene, consistent with subclonal development within tumors. Here, we present examples of using well defined cell populations, including single cells, from complex tissue in combination with molecular tools to reveal features involved in skin carcinogenesis. PMID- 15748640 TI - Dose-dependent effects of UVB-induced skin carcinogenesis in hairless p53 knockout mice. AB - Exposure to (solar) UVB radiation gives rise to mutations in the p53 tumor suppressor gene that appear to contribute to the earliest steps in the molecular cascade towards human and murine skin cancer. To examine in more detail the role of p53, we studied UVB-induced carcinogenesis in hairless p53 knock-out mice. The early onset of lymphomas as well as early wasting of mice interfered with the development of skin tumors in p53 null-mice. The induction of skin tumors in the hairless p53+/- mice was accomplished by daily exposure to two different UV-doses of approximately 450 J/m2 and 900 J/m2 from F40 lamps corresponding to a fraction of about 0.4 and 0.8 of the minimal edemal dose. Marked differences in skin carcinogenesis were observed between the p53+/- mice and their wild type littermates. Firstly, at 900 J/m2, tumors developed significantly faster in the heterozygotes than in wild types, whereas at 450 J/m2 there was hardly any difference, suggesting that only at higher damage levels loss of one functional p53 allele is important. Secondly, a large portion (25%) of skin tumors in the heterozygotes were of a more malignant, poorly differentiated variety of squamous cell carcinomas, i.e. spindle cell carcinomas, a tumor type that was rarely observed in daily UV exposed wild type hairless mice. Thirdly, the p53 mutation spectrum in skin tumors in heterozygotes is quite different from that in wild types. Together these results support the notion that a point mutation in the p53 gene impacts skin carcinogenesis quite differently than allelic loss: the former is generally selected for in early stages of skin tumors in wild type mice, whereas the latter enhances tumor development only at high exposure levels (where apoptosis becomes more prevalent) and appears to increase progression (to a higher grade of malignancy) of skin tumors. PMID- 15748641 TI - Cellular and molecular events leading to the development of skin cancer. AB - The transition from a normal cell to a neoplastic cell is a complex process and involves both genetic and epigenetic changes. The process of carcinogenesis begins when the DNA is damaged, which then leads to a cascade of events leading to the development of a tumor. Ultraviolet (UV) radiation causes DNA damage, inflammation, erythema, sunburn, immunosuppression, photoaging, gene mutations, and skin cancer. Upon DNA damage, the p53 tumor suppressor protein undergoes phosphorylation and translocation to the nucleus and aids in DNA repair or causes apoptosis. Excessive UV exposure overwhelms DNA repair mechanisms leading to induction of p53 mutations and loss of Fas-FasL interaction. Keratinocytes carrying p53 mutations acquire a growth advantage by virtue of their increased resistance to apoptosis. Thus, resistance to cell death is a key event in photocarcinogenesis and conversely, elimination of cells containing excessive UV induced DNA damage is a key step in protecting against skin cancer development. Apoptosis-resistant keratinocytes undergo clonal expansion that eventually leads to formation of actinic keratoses and squamous cell carcinomas. In this article, we will review some of the cellular and molecular mechanisms involved in initiation and progression of UV-induced skin cancer. PMID- 15748642 TI - Inflammation, gene mutation and photoimmunosuppression in response to UVR-induced oxidative damage contributes to photocarcinogenesis. AB - Ultraviolet (UV) radiation causes inflammation, gene mutation and immunosuppression in the skin. These biological changes are responsible for photocarcinogenesis. UV radiation in sunlight is divided into two wavebands, UVB and UVA, both of which contribute to these biological changes, and therefore probably to skin cancer in humans and animal models. Oxidative damage caused by UV contributes to inflammation, gene mutation and immunosuppression. This article reviews evidence for the hypothesis that UV oxidative damage to these processes contributes to photocarcinogenesis. UVA makes a larger impact on oxidative stress in the skin than UVB by inducing reactive oxygen and nitrogen species which damage DNA, protein and lipids and which also lead to NAD+ depletion, and therefore energy loss from the cell. Lipid peroxidation induces prostaglandin production that in association with UV-induced nitric oxide production causes inflammation. Inflammation drives benign human solar keratosis (SK) to undergo malignant conversion into squamous cell carcinoma (SCC) probably because the inflammatory cells produce reactive oxygen species, thus increasing oxidative damage to DNA and the immune system. Reactive oxygen or nitrogen appears to cause the increase in mutational burden as SK progress into SCC in humans. UVA is particularly important in causing immunosuppression in both humans and mice, and UV lipid peroxidation induced prostaglandin production and UV activation of nitric oxide synthase is important mediators of this event. Other immunosuppressive events are likely to be initiated by UV oxidative stress. Antioxidants have also been shown to reduce photocarcinogenesis. While most of this evidence comes from studies in mice, there is supporting evidence in humans that UV-induced oxidative damage contributes to inflammation, gene mutation and immunosuppression. Available evidence implicates oxidative damage as an important contributor to sunlight-induced carcinogenesis in humans. PMID- 15748643 TI - Melanogenesis: a photoprotective response to DNA damage? AB - Exposure to ultra violet radiation (UVR) is associated with significant long-term deleterious effects such as skin cancer. A well-recognised short-term consequence of UVR is increased skin pigmentation. Pigmentation, whether constitutive or facultative, has widely been viewed as photoprotective, largely because darkly pigmented skin is at a lower risk of photocarcinogenesis than fair skin. Research is increasingly suggesting that the relationship between pigmentation and photoprotection may be far more complex than previously assumed. For example, photoprotection against erythema and DNA damage has been shown to be independent of level of induced pigmentation in human white skin types. Growing evidence now suggests that UVR induced DNA photodamage, and its repair is one of the signals that stimulates melanogenesis and studies suggest that repeated exposure in skin type IV results in faster DNA repair in comparison to skin type II. These findings suggest that tanning may be a measure of inducible DNA repair capacity, and it is this rather than pigment per se which results in the lower incidence skin cancer observed in darker skinned individuals. This evokes the notion that epidermal pigmentation may in fact be the mammalian equivalent of a bacterial SOS response. Skin colour is one of most conspicuous ways in which humans vary yet the function of melanin remains controversial. Greater understanding of the role of pigmentation in skin is vital if one is to be able to give accurate advice to the general public about both the population at risk of skin carcinogenesis and also public perceptions of a tan as being healthy. PMID- 15748644 TI - MC1R and the response of melanocytes to ultraviolet radiation. AB - The constitutive color of our skin plays a dramatic role in our photoprotection from solar ultraviolet radiation (UVR) that reaches the Earth and in minimizing DNA damage that gives rise to skin cancer. More than 120 genes have been identified and shown to regulate pigmentation, one of the key genes being melanocortin 1 receptor (MC1R) that encodes the melanocortin 1 receptor (MC1R), a seven-transmembrane G protein-coupled receptor expressed on the surface of melanocytes. Modulation of MC1R function regulates melanin synthesis by melanocytes qualitatively and quantitatively. The MC1R is regulated by the physiological agonists alpha-melanocyte-stimulating hormone (alphaMSH) and adrenocorticotropic hormone (ACTH), and antagonist agouti signaling protein (ASP). Activation of the MC1R by binding of an agonist stimulates the synthesis of eumelanin primarily via activation of adenylate cyclase. The significance of cutaneous pigmentation lies in the photoprotective effect of melanin, particularly eumelanin, against sun-induced carcinogenesis. Epidermal melanocytes and keratinocytes respond to UVR by increasing their expression of alphaMSH and ACTH, which up-regulate the expression of MC1R, and consequently enhance the response of melanocytes to melanocortins. Constitutive skin pigmentation dramatically affects the incidence of skin cancer. The pigmentary phenotype characterized by red hair, fair complexion, inability to tan and tendency to freckle is an independent risk factor for all skin cancers, including melanoma. The MC1R gene is highly polymorphic in human populations, and allelic variation at this locus accounts, to a large extent, for the variation in pigmentary phenotypes and skin phototypes (SPT) in humans. Several allelic variants of the MC1R gene are associated with the red hair and fair skin (RHC) phenotype, and carrying one of these variants is thought to diminish the ability of the epidermis to respond to DNA damage elicited by UVR. The MC1R gene is considered a melanoma susceptibility gene, and its significance in determining the risk for skin cancer is of tremendous interest. PMID- 15748645 TI - Photochemoprevention of ultraviolet B signaling and photocarcinogenesis. AB - Exposure to solar radiation, particularly its ultraviolet (UV) B component, has a variety of harmful effects on human health. Some of these effects include sunburn cell formation, basal and squamous cell cancers, melanoma, cataracts, photoaging of the skin, and immune suppression. Amongst these various adverse effects of UV radiation, skin cancer is of the greatest concern. Over the years, changes in lifestyle has led to a significant increase in the amount of UV radiation that people receive, and this consequently has led to a surge in the incidence of skin cancer. The development of skin cancer is a complex multistage phenomenon involving three distinct stages exemplified by initiation, promotion and progression stages. Each of these stages is mediated via alterations in various cellular, biochemical, and molecular changes. Initiation, the first step in the carcinogenesis process is essentially an irreversible step in which genetic alterations occur in genes that ultimately leads to DNA modification and fixation of mutation. Tumor promotion is the essential process in cancer development involving clonal expansion of initiated cells giving rise to pre-malignant and then to malignant lesions, essentially by alterations in signal transduction pathways. Tumor progression involves the conversion of pre-malignant and malignant lesions into an invasive and potentially metastatic malignant tumor. All these processes for skin cancer development involve stimulation of DNA synthesis, DNA damage and proliferation, inflammation, immunosuppression, epidermal hyperplasia, cell cycle dysregulation, depletion of antioxidant defenses, impairment of signal transduction pathways, induction of cyclooxygenase, increase in prostaglandin synthesis, and induction of ornithine decarboxylase. Photochemoprevention has been appreciated as a viable approach to reduce the occurrence of skin cancer and in recent years, the use of agents, especially botanical antioxidants, present in the common diet and beverages consumed by human population have gained considerable attention as photochemopreventive agents for human use. Many such agents have also found a place in skin care products. Although this is more common in oriental countries, its popularity is significantly growing in western countries. In this article, we have summarized the available information of laboratory studies on UVB-mediated signaling that can be exploited as targets for photochemoprevention. We suggest that the use of skin care products supplemented with proven chemopreventive agents in conjunction with the use of sunscreens along with educational efforts may be an effective strategy for reducing UV-induced photodamage and skin cancer in humans. The mechanistic basis for the use of such products is discussed. PMID- 15748646 TI - Importance of UVA photoprotection as shown by genotoxic related endpoints: DNA damage and p53 status. AB - In order to demonstrate the importance of photoprotection in the UVA range (320 400 nm), an in vitro approach where sun formulations are spread on a quartz slide, and placed over human keratinocytes in culture is proposed as a convenient test for photoprotection assessment at the DNA level. Using the comet assay, DNA strand breaks, oxidative DNA damage or drug-induced DNA breaks were assessed. Accumulation of p53 protein was also studied as a marker for UV-induced genotoxic stress. Such a method was used to compare two formulations with different photostability. Spectroradiometry showed that a photounstable formulation lost its effectiveness in UVA screening when pre-irradiated by simulated sunlight (UVB+UVA). As a consequence, it was also shown that this formulation was not as protective as the photostable one at the genomic level. These data demonstrate that the loss of absorbing efficiency within UVA wavelengths due to photounstability may have detrimental consequences leading to impairments implicated in genotoxic events. PMID- 15748647 TI - Mechanisms underlying UV-induced immune suppression. AB - Skin cancer is the most prevalent form of human neoplasia. Estimates suggest that in excess of one million new cases of skin cancer will be diagnosed this year alone in the United States (www.cancer.org/statistics). Fortunately, because of their highly visible location, skin cancers are more rapidly diagnosed and more easily treated than other types of cancer. Be that as it may, approximately 10,000 Americans a year die from skin cancer. The cost of treating non-melanoma skin cancer is estimated to be in excess of US dollars 650 million a year, and when melanoma is included, the estimated cost of treating skin cancer in the United States is estimated to rise to US dollars 2.9 billion annually (www.cancer.org/statistics). Because the morbidity and mortality associated with skin cancer is a major public health problem, it is important to understand the mechanisms underlying skin cancer development. The primary cause of skin cancer is the ultraviolet (UV) radiation found in sunlight. In addition to its carcinogenic potential, UV radiation is also immune suppressive. In fact, data from studies with both experimental animals and biopsy proven skin cancer patients suggest that there is an association between the immune suppressive effects of UV radiation and its carcinogenic potential. The focus of this manuscript will be to review the mechanisms underlying the induction of immune suppression following UV exposure. Particular attention will be directed to the role of soluble mediators in activating immune suppression. PMID- 15748648 TI - Ultraviolet radiation: effects on risks of prostate cancer and other internal cancers. AB - Governmental and research agencies worldwide have strongly advocated sun avoidance strategies in an attempt to counter marked increases in skin cancer incidence. Concurrently, there are reports describing widespread Vitamin D3 deficiency. Because 1,25-dihydroxyvitamin D3, through interaction with the Vitamin D receptor, exerts pleiotrophic effects, such deficiency might be expected to have clinical consequences. Indeed, various reports indicate that exposure to ultraviolet radiation (UVR) exerts a protective effect on development of some common diseases including internal cancers and multiple sclerosis. We describe studies indicating that modest exposure reduces risk of prostate cancer. The effect of UVR is mediated by skin type; at lower levels of exposure a relative inability to effect skin pigmentation is protective presumably because it allows more efficient Vitamin D3 synthesis. Polymorphic variants in genes associated with pigmentation including melanocyte stimulating hormone receptor and tyrosinase are also associated with prostate cancer risk. Overall, though preliminary and requiring cautious interpretation, these data indicate that moderate UVR exposure together with characteristics linked with less effective tanning confer reduced prostate cancer risk. Clearly, it is important to define safe levels of UVR that do not result in increased risk of skin cancers such as malignant melanoma. PMID- 15748649 TI - Solar ultraviolet radiation-induced DNA damage in aquatic organisms: potential environmental impact. AB - Continuing depletion of stratospheric ozone and subsequent increases in deleterious ultraviolet (UV) radiation at the Earth's surface have fueled the interest in its ecological consequences for aquatic ecosystems. The DNA is certainly one of the key targets for UV-induced damage in a variety of aquatic organisms. UV radiation induces two of the most abundant mutagenic and cytotoxic DNA lesions, cyclobutane pyrimidine dimers (CPDs) and pyrimidine pyrimidone photoproducts (6-4PPs) and their Dewar valence isomers. However, aquatic organisms have developed a number of repair and tolerance mechanisms to counteract the damaging effects of UV on DNA. Photoreactivation with the help of the enzyme photolyase is one of the most important and frequently occurring repair mechanisms in a variety of organisms. Excision repair, which can be distinguished into base excision repair (BER) and nucleotide excision repair (NER), also play an important role in DNA repair in several organisms with the help of a number of glycosylases and polymerases, respectively. In addition, mechanisms such as mutagenic repair or dimer bypass, recombinational repair, cell cycle checkpoints, apoptosis and certain alternative repair pathways are also operative in various organisms. This review deals with the UV-induced DNA damage and repair in a number of aquatic organisms as well as methods of detecting DNA damage. PMID- 15748650 TI - Dynamic response of plant genome to ultraviolet radiation and other genotoxic stresses. AB - Oligonucleotide microarray technology was used to identify genes, which are responding after exposure to UV-C radiation and to other agents causing genotoxic stress. The effect of these conditions on recombinational DNA repair was monitored in parallel. Global changes in gene expression were investigated in Arabidopsis wild-type plants challenged with UV-C, bleomycin, another abiotic agent and xylanase, a biotic factor, all leading to elevated homologous recombination frequencies. The comparison of the expression profile of each treatment allowed defining genes specifically involved in the dynamic response to UV. In the future, the potential roles of such genes in the different forms of stress recognition, signal transduction, and their roles in DNA repair processes will be assessed by using reverse genetic tools available for Arabidopsis thaliana. PMID- 15748651 TI - The solar UV environment and bacterial spore UV resistance: considerations for Earth-to-Mars transport by natural processes and human spaceflight. AB - The environment in space and on planets such as Mars can be lethal to microorganisms because of the high vacuum and high solar radiation flux, in particular UV radiation, in such environments. Spores of various Bacillus species are among the organisms most resistant to the lethal effects of high vacuum and UV radiation, and as a consequence are of major concern for planetary contamination via unmanned spacecraft or even natural processes. This review focuses on the spores of various Bacillus species: (i) their mechanisms of UV resistance; (ii) their survival in unmanned spacecraft, space flight and simulated space flight and Martian conditions; (iii) the UV flux in space and on Mars; (iv) factors affecting spore survival in such high UV flux environments. PMID- 15748652 TI - Damage and repair of ancient DNA. AB - Under certain conditions small amounts of DNA can survive for long periods of time and can be used as polymerase chain reaction (PCR) substrates for the study of phylogenetic relationships and population genetics of extinct plants and animals, including hominids. Because of extensive DNA degradation, these studies are limited to species that lived within the past 10(4)-10(5) years (Late Pleistocene), although DNA sequences from 10(6) years have been reported. Ancient DNA (aDNA) has been used to study phylogenetic relationships of protists, fungi, algae, plants, and higher eukaryotes such as extinct horses, cave bears, the marsupial wolf, the moa, and Neanderthal. In the past few years, this technology has been extended to the study of infectious disease in ancient Egyptian and South American mummies, the dietary habits of ancient animals, and agricultural practices and population dynamics of early native Americans. Hence, ancient DNA contains information pertinent to numerous fields of study including evolution, population genetics, ecology, climatology, medicine, archeology, and behavior. The major obstacles to the study of aDNA are its extremely low yield, contamination with modern DNA, and extensive degradation. In the course of this review, we will discuss the current aDNA literature describing the importance of aDNA studies as they relate to important biological questions and the difficulties associated with extracting useful information from highly degraded and damaged substrates derived from limited sources. In addition, we will present some of our own preliminary and published data on mechanisms of DNA degradation and some speculative thoughts on strategies for repair and restoration of aDNA. PMID- 15748653 TI - Epoxide hydrolases: their roles and interactions with lipid metabolism. AB - The epoxide hydrolases (EHs) are enzymes present in all living organisms, which transform epoxide containing lipids by the addition of water. In plants and animals, many of these lipid substrates have potent biologically activities, such as host defenses, control of development, regulation of inflammation and blood pressure. Thus the EHs have important and diverse biological roles with profound effects on the physiological state of the host organisms. Currently, seven distinct epoxide hydrolase sub-types are recognized in higher organisms. These include the plant soluble EHs, the mammalian soluble epoxide hydrolase, the hepoxilin hydrolase, leukotriene A4 hydrolase, the microsomal epoxide hydrolase, and the insect juvenile hormone epoxide hydrolase. While our understanding of these enzymes has progressed at different rates, here we discuss the current state of knowledge for each of these enzymes, along with a distillation of our current understanding of their endogenous roles. By reviewing the entire enzyme class together, both commonalities and discrepancies in our understanding are highlighted and important directions for future research pertaining to these enzymes are indicated. PMID- 15748654 TI - Multiple forms of secretory phospholipase A2 in plants. AB - Multiple secretory phospholipase A2 (sPLA2) genes have been identified in plants and encode isoforms with distinct regulatory and catalytic properties. Elucidation of this genetic and biochemical heterogeneity has provided important clues to the regulation and function of the individual enzymes. An increasing body of evidence shows that their lipid products, lysophospholipids and free fatty acids, mediate a variety of cellular responses, including plant growth, development, and responses to stress and defense. This review discusses the newly acquired information on plant sPLA2s including the molecular and biochemical characteristics, and signaling functions of each isoform. PMID- 15748655 TI - Advanced strategies in liposomal cancer therapy: problems and prospects of active and tumor specific drug release. AB - Tumor specific drug delivery has become increasingly interesting in cancer therapy, as the use of chemotherapeutics is often limited due to severe side effects. Conventional drug delivery systems have shown low efficiency and a continuous search for more advanced drug delivery principles is therefore of great importance. In the first part of this review, we present current strategies in the drug delivery field, focusing on site-specific triggered drug release from liposomes in cancerous tissue. Currently marketed drug delivery systems lack the ability to actively release the carried drug and rely on passive diffusion or slow non-specific degradation of the liposomal carrier. To obtain elevated tumor to-normal tissue drug ratios, it is important to develop drug delivery strategies where the liposomal carriers are actively degraded specifically in the tumor tissue. Many promising strategies have emerged ranging from externally triggered light- and thermosensitive liposomes to receptor targeted, pH- and enzymatically triggered liposomes relying on an endogenous trigger mechanism in the cancerous tissue. However, even though several of these strategies were introduced three decades ago, none of them have yet led to marketed drugs and are still far from achieving this goal. The most advanced and prospective technologies are probably the prodrug strategies where non-toxic drugs are carried and activated specifically in the malignant tissue by overexpressed enzymes. In the second part of this paper, we review our own work, exploiting secretory phospholipase A2 as a site-specific trigger and prodrug activator in cancer therapy. We present novel prodrug lipids together with biophysical investigations of liposome systems, constituted by these new lipids and demonstrate their degradability by secretory phospholipase A2. We furthermore give examples of the biological performance of the enzymatically degradable liposomes as advanced drug delivery systems. PMID- 15748656 TI - Design of a radiation surveillance unit for an unmanned aerial vehicle. AB - This paper describes a prototype of a compact environmental radiation surveillance instrument designed for a Ranger unmanned aerial vehicle. The instrument, which can be used for tracking a radioactive plume, mapping fallout and searching for point sources, consists of three different detector types (GM, NaI(Tl) and CZT) and an air sampling unit. In addition to the standard electronics for data acquisition, the system contains an onboard computer, a GPS receiver and environmental sensors, all enclosed in a single housing manufactured of fiberglass-reinforced composite material. The data collected during the flight is transmitted in real-time to the ground station via a TETRA radio network. The radiation surveillance unit is an independent module and as such can be used in, for example, airplanes, helicopters and cars. PMID- 15748657 TI - Natural radioactivity in sand used in thermal therapy at the Red Sea Coast. AB - The development of climatotherapy in Safaga opens the field of medical tourism in Egypt, in order to detect any harmful radiation that would affect the patients during treatment and is becoming important economic resource. Studies and survey of natural radiation and radioactivity in upper Egypt conducted since 1990, included monitoring of the concentration of natural radionuclides in environmental samples. The results of the study reveals that, for all sand samples, the mean activity concentration of 40K (618+/-122-548+/-82 Bq kg(-1)) are much higher than that of both 226Ra (25.3+/-14-20.6+/-10 Bq kg(-1)) and 232Th (21.4+/-10-22.4+/-10 Bq kg(-1)). Different radiation hazard indices were calculated, the radiation dose to which workers are subjected is not negligible (26.5-50.9 nGy h(-1)), although depending on the inhalation of dust. PMID- 15748658 TI - Characterization of uranium and plutonium containing particles originating from the nuclear weapons accident in Thule, Greenland, 1968. AB - To improve long-term radioecological impact assessment for the contaminated ecosystem of Bylot Sound, Greenland, U and Pu containing particles have been characterized with respect to particle size, elemental distribution, morphology and oxidation states. Based on scanning electron microscopy with XRMA, particles ranging from about 20 to 40 microm were isolated. XRMA and mu-XRF mapping demonstrated that U and Pu were homogeneously distributed throughout the particles, indicating that U and Pu have been fused. Furthermore, mu-XANES showed that U and Pu in the particles were present as mixed oxides. U was found to be in oxidation state IV whereas Pu apparently is a mixture of Pu(III) and Pu(IV). As previous assessments are based on PuO2 only, revisions should be made, taking Pu(III) into account. PMID- 15748659 TI - Accumulation and release of 241Am by a macrophyte of the Yenisei River (Elodea canadensis). AB - The source of radioactive contamination of the Yenisei River floodplain, including contamination with transuranic elements, is the Mining-and-Chemical Combine of the Russian Ministry of Atomic Energy, which has for many years been producing weapons-grade plutonium. Transuranic elements have been detected not only in the soil and sediment of the river but also in the biomass of aquatic plants. This work is an investigation of accumulation and release of 241Am by a submerged macrophyte of the Yenisei River (Elodea canadensis) in laboratory experiments. In 2000-2003, laboratory experiments were carried out with biomass of E. canadensis Mich. and filtered river water. The samples were collected from the Yenisei River upstream of the discharge of the Combine's radioactive effluent. The experiments showed that 241Am is accumulated by Elodea biomass: the activity concentration of 241Am can reach 3280+/-240 Bq/g, with the concentration factor for 241Am 16 600+/-2200l/kg. Results of chemical fractionation have proved that in the course of 241Am accumulation by Elodea biomass, 241Am tightly bound to biomass increases from 11% to 27% of the total 241Am in the biomass. Release of 241Am from the decaying Elodea biomass has been evaluated experimentally. By the end of the experiment (lasting up to 127 days), the Elodea plants had lost up to 65% of their initial 241Am activity and the rate of 241Am release into the water environment reached 23 Bq/day. PMID- 15748660 TI - Radioactivity concentration in liquid and solid phases of scale and sludge generated in the petroleum industry. AB - Scales and sludge generated during oil extraction and production can contain uranium, thorium, radium and other natural radionuclides, which can cause exposure of maintenance personnel. This work shows how the oil content can influence the results of measurements of radionuclide concentration in scale and sludge. Samples were taken from a PETROBRAS unit in Northeast Brazil. They were collected directly from the inner surface of water pipes or from barrels stored in the waste storage area of the E&P unit. The oil was separated from the solids with a Soxhlet extractor by using aguarras at 90+/-5 degrees C as solvent. Concentrations of 226Ra and 228Ra in the samples were determined before and after oil extraction by using an HPGe gamma spectrometric system. The results showed an increase in the radionuclide concentration in the solid (dry) phase, indicating that the above radionuclides concentrate mostly in the solid material. PMID- 15748661 TI - 137Cs concentration in meat of wild boars (Sus scrofa) in Croatia a decade and half after the Chernobyl accident. AB - Gamma-spectrometric measurements of 137Cs activities in meat of wild boars collected in Croatia at several locations with different levels of 137Cs contamination are presented. Samples were collected during the period between 2000 and 2002, about 15 years after the Chernobyl accident. 137Cs concentrations ranged over three orders of magnitude: 0.4-611.5 Bq kg(-1). On the basis of these results, 137Cs concentrations at researched areas could be categorized into three groups: (i) the area of Slavonski Brod, Lipik and Slunj with 137Cs concentrations in meat of only a few Bq kg(-1); (ii) the area of Vrbovsko and Sirac with 137Cs concentrations of a few tens of Bq kg(-1); and (iii) the Fuzine area with 137Cs values in wild boar meat of a few hundreds of Bq kg(-1). In areas with approximately equal contamination level, 137Cs concentrations in wild boar meat varied over two orders of magnitude. This fact suggests that the main reason for high 137Cs values in wild boar meat could be due to food consumed by wild boars, and only secondarily in contamination level of area where they live. Intensive mushroom consumption during autumn months could be one of the factors responsible for high 137Cs values in wild boar meat. An average dose arising from 137Cs due to ingestion of wild boar meat in Croatia is below radiological health concern except in the area of Fuzine, and only in cases of high annual wild boar meat intake, probably by hunters or members of their families. PMID- 15748662 TI - 90Sr, 137Cs and (239,240)Pu concentration surface water time series in the Pacific and Indian Oceans--WOMARS results. AB - Under an IAEA's Co-ordinated Research Project "Worldwide Marine Radioactivity Studies (WOMARS)" 90Sr, 137Cs and (239,240)Pu concentration surface water time series in the Pacific and Indian Oceans have been investigated. The Pacific and Indian Oceans were divided into 17 latitudinal boxes according to ocean circulation, global fallout patterns and the location of nuclear weapons test sites. The present levels and time trends in radionuclide concentrations in surface water for each box were studied and the corresponding effective half lives were estimated. For the year 2000, the estimated average 90Sr, 137Cs and (239,240)Pu concentrations in surface waters of the Pacific and Indian Oceans varied from 0.1 to 1.5 mBq/L, 0.1 to 2.8 mBq/L, and 0.1 to 5.2 microBq/L, respectively. The mean effective half-lives for 90Sr and 137Cs in surface water were 12+/-1 years for the North, 20+/-1 years for the South and 21+/-2 years for the Equatorial Pacific. For (239,240)Pu the corresponding mean effective half lives were 7+/-1 years for the North, 12+/-4 years for the South and 10+/-2 years for the Equatorial Pacific. For the Indian Ocean the mean effective half-lives of 137Cs and (239,240)Pu were 21+/-2 years and 9+/-1 years, respectively. There is evidence that fallout removal rates before 1970 were faster than those observed during recent decades. The estimated surface water concentrations of 90Sr, 137Cs and (239,240)Pu in latitudinal belts of the Pacific and Indian Oceans for the year 2000 may be used as the average levels so that any new contribution from nuclear facilities, nuclear weapons test sites, radioactive waste dumping sites and from possible nuclear accidents can be identified. PMID- 15748663 TI - Soil-fungi transfer coefficients: Importance of the location of mycelium in soil and of the differential availability of radionuclides in soil fractions. AB - Soil-fungus transfer coefficients are usually defined as the ratio between the content of the fruiting bodies and that of the soil. Since, however, the methodology of how to determine the soil content is not firmly established, there exist a variety of definitions in the literature. We analyzed the 137Cs, 90Sr, 40K, and 226Ra content of mushroom and soil samples from two pine-wood ecosystems in Spain. The location of the mycelium in the soil profiles of these ecosystems was determined by means of the ergosterol concentration. The results showed the mycelium to generally be localized in the surface layer of soil (0-5 cm). We also carried out a speciation procedure for this layer of soil to determine the different degrees of association of the radionuclides in the soil. The results led us to propose some variations to the traditional definition used in quantifying radionuclide transfer. With these modifications, we were able to analyze Cs-K competition in several species of mycorrhizal and saprophytic fungi. PMID- 15748664 TI - Radionuclide content of concrete building blocks and radiation dose rates in some dwellings in Ibadan, Nigeria. AB - Thirty-two samples of concrete building blocks were collected from different block making industries in Ibadan. The radioactivity concentrations of the natural radionuclides in the samples were determined by gamma-ray spectrometry with a NaI(Tl) detector. The radioactivity concentrations varied from 6.2 to 57.5 Bq kg(-1), 12.4 to 64.9 Bq kg(-1) and 95.3 to 766.1 Bq kg(-1) for 226Ra, 232Th and 40K, respectively. The radium equivalent activities of the 32 samples varied from 51.3 to 175.7 Bq kg(-1). Radiation exposure levels in 30 dwellings were determined using LiF thermoluminescent dosimeters. The annual equivalent dose rates varied from 0.318 to 0.657 mSv y(-1) with a mean of 0.433 mSv y(-1). The annual effective dose rate to the whole body was calculated as 0.236 mSv y(-1), which is less than that (mean) estimated by UNSCEAR for normal background areas. PMID- 15748666 TI - Canadian Aboriginal people's experiences with HIV/AIDS as portrayed in selected English language Aboriginal media (1996-2000). AB - This paper describes the portrayal of HIV/AIDS in 14 mass print newspapers directed towards the Canadian Aboriginal population and published between 1996 and 2000. Based on qualitative content analysis the research examines both manifest and latent meanings. Manifest results of this study indicate that women and youth are under represented as persons with HIV/AIDS. The latent results note the frequent references to Aboriginal culture, and the political and economic position of Aboriginal Canadians when discussing the disease, the person with the disease, the fear of the disease and the reaction of the community to the person with the disease. Unlike mainstream media where the medical frame is dominant, HIV/AIDS are here contextualized by culture, identity, spirituality and political economic issues. PMID- 15748665 TI - Extended family's and women's roles in safeguarding orphans' education in AIDS afflicted rural Zimbabwe. AB - The extended family forms the basis for orphan care and education in sub-Saharan Africa. Initial absence followed by emergence of differentials in primary school enrollment between orphans and non-orphans have been attributed to the strength and subsequent HIV/AIDS-induced breakdown of extended family orphan care arrangements. Yet, few attempts have been made to describe how these arrangements are affected by HIV/AIDS or how they relate to observed patterns of childhood outcomes by sex and orphan status. We use a combination of quantitative and qualitative data to show that maternal orphans but not paternal or double orphans have lower primary school completion rates than non-orphans in rural Zimbabwe, and that these patterns reflect adaptations and gaps in extended family orphan care arrangements. Sustained high levels of primary school completion amongst paternal and double orphans--particularly for girls--result from increased residence in female-headed households and greater access to external resources. Low primary school completion amongst maternal orphans results from lack of support from fathers and stepmothers and ineligibility for welfare assistance due to residence in higher socio-economic status households. These effects are partially offset by increased assistance from maternal relatives. These findings indicate that programmes should assist maternal orphans and support women's efforts by reinforcing the roles of extended families and local communities, and by facilitating greater self-sufficiency. PMID- 15748667 TI - Commercial sex work, drug use, and sexually transmitted infections in St. Petersburg, Russia. AB - The relationships between commercial sex work, drug use, and sexually transmitted infections (STI) in St. Petersburg, Russia were assessed using qualitative research methods and an examination of existing research, surveillance and epidemiology data. The rapid assessment methodology included in-depth qualitative interviews with key informants, naturalistic observations of commercial sex work and drug use sites, geo-mapping, and a critical review of the available surveillance, epidemiology, and sociological data. Patterns of commercial sex work and drug use in St. Petersburg are described. The existing surveillance data attributes infections to injected drug use over and above any other risk category. However, examination of the clinic and epidemiology data suggests that HIV infection may be increasing fastest among groups that are acquiring HIV through sexual transmission. Targeted screening studies of STI and HIV morbidity among populations that are not included in the surveillance algorithm are needed, such as commercial sex workers, street youth, and the homeless. Sexual history taking to better characterize the proportion of cases that result from sex between male partners would also be helpful. PMID- 15748668 TI - What girls need: recommendations for preventing violence among urban girls in the US. AB - The last decade saw increases in arrests of girls for violent behavior and a corresponding concern that girls' involvement in violence was increasing in the USA. However, there are few empirical studies of the dynamics of violence by girls, leaving providers of violence prevention programs and policy-makers without evidence on which to base gender-appropriate prevention strategies. To address this gap, qualitative interviews were conducted with a diverse sample of 61 urban girls aged 11-17. Findings were compared with quantitative interviews from the prospective cohort of 961 girls from whom these respondents were drawn, from the Project on Human Development in Chicago Neighborhoods. Mixed-method techniques were employed. Qualitative data were analyzed for girls' recommendations for preventing involvement in violence. Data from the larger cohort were used to test these recommendations quantitatively. Due to study design, in the qualitative sample, 36 girls (64%) were involved in recent violence, most often with or against other girls. Pro-social behavior was common among both violent and nonviolent girls. In the overall cohort sample, 24.9% of girls reported violent perpetration and 97% reported pro-social activities. Eight themes regarding staying safe and preventing violence emerged from the qualitative interviews: girls stayed safe by staying home, avoiding dangerous people, staying busy with after-school activities, remaining calm when confronted, using escorts, and fighting back if attacked. Girls' protective influences included: empathic parental involvement, positive relationships with peers and older youth, and involvement in safe and constructive activities. These findings emphasize that safety in community, school, and family settings is critical for girls in avoiding violence and other risky behaviors. Violence prevention programs should focus on enhancing girls' relationships with mothers, older girls, and friends their age. PMID- 15748669 TI - Community level effects of gender inequality on intimate partner violence and unintended pregnancy in Colombia: testing the feminist perspective. AB - Violence against women, especially by intimate partners, is a serious public health problem that is associated with physical, reproductive, and mental health consequences. The effect of intimate partner violence on women's ability to control their fertility and the mechanisms through which these phenomena are related merit further investigation. Building on findings from a previous analysis in which a statistically significant relationship between intimate partner violence and unintended pregnancy in Colombia was found, this analysis examines the effect of gender inequality on this association using data from the 2000 Colombian Demographic and Health Survey. Specifically, the objective of this analysis is to explore whether gender inequality (as measured by women's autonomy, women's status, male patriarchal control, and intimate partner violence) in municipalities partially explains the association between intimate partner violence and unintended pregnancy in Colombia. Results of logistic regression analysis with multi-level data show that living in a municipality with high rates of male patriarchal control significantly increased women's odds of having an unintended pregnancy by almost four times. Also, living in a municipality with high rates of intimate partner violence increased one's odds of unintended pregnancy by more than 2.5 times, and non-abused women living in municipalities with high rates of intimate partner violence were at a significantly increased risk of unintended pregnancy. In addition, abused women living in a municipality with high personal female decision-making autonomy had more than a fourfold increased risk of having an unintended pregnancy. These findings demonstrate the need for reproductive health programs to target areas at particularly high risk for unintended pregnancy by reducing intimate partner violence and gender inequality. PMID- 15748670 TI - Race and ethnic differences in determinants of preterm birth in the USA: broadening the social context. AB - Preterm births occur in 9.7% of all US singleton births. The rate for blacks is double that of whites and the rate is 25% higher for Hispanics than for whites. While a number of individual correlates with preterm birth have been identified, race and ethnic differences have not been fully explained. Influenced by a growing body of literature documenting a relationship among health, individual income, and neighborhood disadvantage, researchers interested in explaining racial differences in preterm birth are designing studies that extend beyond the individual. No studies of adverse birth outcomes have considered contextual effects beyond the neighborhood level. Only a handful of studies, comparing blacks and whites, have evaluated the influence of neighborhood disadvantage on preterm birth. This study examines how preterm birth among blacks, whites and Hispanics is influenced by social context, broadly defined to include measures of neighborhood disadvantage and cumulative exposure to state-level income inequality, controlling for individual risk factors. Neighborhood disadvantage is determined by Census tract data. Cumulative exposure to income inequality is measured by the fraction of the mother's life since age 14 spent residing in states with a state-level Gini coefficient above the median. The results for neighborhood disadvantage are highly sensitive across race/ethnicities to the measure used. We find evidence that neighborhood poverty rates and housing vacancy rates increased the rate of very preterm birth and decreased the rate of moderately preterm birth for blacks. The rate of very preterm increased with the fraction of female-headed households for Hispanics and decreased with the fraction of people employed in professional occupations for whites. We find direct effects of cumulative exposure to income inequality only for Hispanics. However, we do find indirect effects of context broadly defined on behaviors that increased the risk of preterm birth. PMID- 15748671 TI - The effects of racial density and income incongruity on pregnancy outcomes. AB - This study shows that living in a better area reduces the risk of adverse pregnancy outcomes but, among African-American women, living in an area in which they are in a racial minority may increase the risk. Using the 1991 cohort of single infants born to African-American women in Chicago, we measured census tract socioeconomic status and defined women as having "positive income incongruity" if they lived in wealthier tracts than the average African-American woman of comparable education and marital status. We examined whether or not the effect of positive income incongruity differed according to whether or not African-American women lived in predominantly black, or mixed tracts. Among the women living in predominantly black census tracts, positive income incongruity was associated with a lower risk of low birth weight (odds ratio (OR)=0.91) and preterm delivery (OR=0.83). These effects were modest, but statistically significant for gestation (p-value=0.01). In contrast, among the women living in mixed tracts positive income incongruity was not associated with low birth weight (OR=1.04) or preterm delivery (OR=1.11). In mixed areas the expected benefits of positive income incongruity are completely offset by the racial density effect, suggesting that the positive effects of a better socioeconomic context may be countered for minority women by the adverse effects of racism or racial stigma. PMID- 15748672 TI - Does chronic illness place constraints on positive constructions of identity? Temporal comparisons and self-evaluations in people with schizophrenia. AB - It is often suggested that people in potentially threatening situations might engage in self-enhancing temporal comparisons that allow them to view themselves and their experience in a more positive light. Data from semi-structured interviews with 12 individuals in the UK diagnosed as having schizophrenia were content analyzed to explore patterns of temporal comparison. The study found that the onset of schizophrenic symptoms created a new baseline in participants' representations of their past, with different types of temporal comparisons occurring before and after this point. Although comparisons with past selves after onset supported the suggestion that people may select and construct their past in such a manner that permits them to see their present circumstances more positively and envisage a better future, comparisons with past selves before onset were more negative. The findings suggest that the Theory of Temporal Self Appraisals (Ross & Wilson, 2000) needs to be elaborated to include people who have experienced major life changes. PMID- 15748673 TI - Constructions of masculinity following prostatectomy-induced impotence. AB - Large numbers of Australian men are diagnosed and treated for prostate cancer each year. The incidence is exceeding mortality, and men are living longer with prostate cancer and the common treatment[s] side effect of impotence. Despite these epidemiological trends there is little research about men's experiences of impotence following treatment. An ethnographic study of Anglo-Australian men with localized prostate cancer explored participants' experiences of impotence following prostatectomy. In-depth semi-structured interviews with 15 men were analyzed using a social constructionist gendered framework. In particular, the effect of impotence on participants' masculinity, sexuality and intimate relationships was explored. The findings show that participants rationalized forgoing potency prior to surgery as a way of living longer. However, diverse complex reactions accompanied impotence. Whilst most participants redefined masculine ideals of phallocentric sex, the way in which this occurred varied greatly. The findings disrupt essentialist constructions of male sexuality and impotence, and provide valuable insight for clinical practice. PMID- 15748674 TI - Sickness absence in female- and male-dominated occupations and workplaces. AB - Previous research suggests that both men's and women's level of sickness absence may be systematically related to the gender composition of their workplace as well as of their occupational category. The number of studies is, however, low and the composition of the occupational category has often been used as a proxy for the composition of the workplace. This paper employs a large data set broadly representative of the employed population of Norway. The data make it possible to take workplace and occupation simultaneously into account. Thus, the relationship between the gender composition of the workplace and sickness absence is estimated with detailed control for differences between occupational categories. Likewise, the importance of the gender composition of the occupation is assessed with control for between workplace variation. Men's sickness absence turns out to be largely unrelated to the gender composition of the workplace. For women the level of sickness absence tends to be higher in female-dominated workplaces, but the relationship is weak. These findings provide evidence against theories suggesting that the minority sex in the workplace faces special problems and is therefore more absent. They are to some extent consistent with the idea that female dominated workplaces develop norms that are more tolerant towards sickness absence. The relationship of sickness absence to the gender composition of the occupational category is similar to the U-shaped pattern found in several previous studies (highest sickness absence both in strongly male-dominated and strongly female-dominated occupations), but again the relationship is weak. PMID- 15748675 TI - 'Choosing' to work when sick: workplace presenteeism. AB - Presenteeism is a concept used to describe the phenomenon of working through illness and injury. This paper is based on interviews and focus groups undertaken at three different work sites in New Zealand: a small private hospital, a large public hospital and a small factory. The research suggests that presenteeism is a prominent phenomenon in the lives of workers at these different sites, but the way in which it is rationalised and the factors that foster presenteeism are quite distinct. Exploring the way in which presenteeism links to economic and social constraints and workplace cultures provides insights into these rationalisations. The powerful forces promoting presenteeism tempers the research community's concern with absenteeism. A presenteeism discourse needs to be more prominently articulated to oppose both the absenteeism discourse, and to moderate the views taken by some postmodernist theorists on choice in relation to health practices in workplace settings. PMID- 15748676 TI - 'Supply' and 'demand': breastfeeding as labour. AB - This paper presents findings from a recent critical ethnographic study conducted in two maternity units in England, UK. The study explored the influences upon 61 women's experiences of breastfeeding within the postnatal ward setting. Participant observations of 97 encounters between midwives and postnatal women, 106 focused interviews with postnatal women and 37-guided conversations with midwives were conducted. Basic, organising and global themes were constructed utilising thematic networks analysis. The metaphor of the production line, with its notions of demand and efficient supply, illustrated the experiences of breastfeeding women. They conceptualised breastfeeding as a 'productive' project, yet expressed deep mistrust in the efficacy of their bodies. Their emphasis centred upon breast milk as nutrition rather than relationality and breastfeeding. Women referred to the demanding and unpredictable ways in which their baby breached their temporal and spatial boundaries. They sought strategies to cope with the uncertainty of this embodied experience in combination with their concerns regarding returning to a 'normal' and 'productive' life. The hospital setting and health worker practices played a contributing and reinforcing role. The paper discusses ways of re-establishing trust in women's bodies and breastfeeding, while respecting difference and diversity. It argues for embracing the concepts of embodiment and relationality whilst avoiding a return to essentialism. This requires collective efforts to erode deeply embedded cultural understandings of women's bodies centering upon disembodied and efficient production. PMID- 15748677 TI - Mothers reframing physical activity: family oriented politicism, transgression and contested expertise in Australia. AB - Mothers of young children are a population sub-group with one of the lowest levels of physical activity. This paper presents the findings from a qualitative study with 40 Australian mothers of children under school age. The research aimed to understand the tensions, dilemmas and trade-offs which women experience around physical activity within the contexts of their everyday lives as mothers of young children. The analysis shows that, in contrast to health promotion messages which frame physical activity as a positive and healthy behaviour, mothers of young children perceive activity as being both enhancing and threatening to their health and social relationships. Restrictive stereotypes of the 'good' mother make it difficult for many women to prioritise their own physical activity needs over their childrearing and domestic responsibilities. Nevertheless, women's involvement in physical activity is often underpinned by the maternal 'ethic of care' as something which can help them cope better with the challenges of being a mother and contribute to the wellbeing of the family. This article takes as its departure point the notion that the maternal 'ethic of care' creates previously unrecognised opportunities in relation to physical activity. For many mothers, physical activity can also be a way of challenging hegemonic discourses and extending what it means to be a good mother in contemporary society. Although largely overlooked by contemporary health promotion, it is women's family oriented politicism and resistance to dominant meanings about motherhood, health and the 'ideal' body which create alternative possibilities for their participation and enjoyment of physical activity during early motherhood. PMID- 15748678 TI - Physician-elderly patient-companion communication and roles of companions in Japanese geriatric encounters. AB - Although the triadic encounter of physician, patient, and an accompanying family member is a common phenomenon in geriatrics, previous research on the communication in medical encounters has primarily focused on dyadic interactions between physician and patient. This study aimed to explore the triadic communication and communication roles of patient companions in Japanese geriatric encounters. Among elderly patients aged 65 or over who were under continuous care of nine attending physicians at a university affiliated geriatric clinic in Tokyo, 63 accompanied patients and 82 unaccompanied patients were included for this study. The consultation was audiotape recorded and analyzed using the Roter Interaction Analysis System (RIAS) with additional categories developed to code aspects of companion communication. In dyadic encounters, the average proportions of physician's talk and patient's talk were 54% and 46%, respectively, while in triadic encounters the average talk proportions of physician, patient, and companion were 49%, 29%, and 22%. Companions made a significant contribution to the communication during the visit by providing information and asking the physician questions, as well as facilitating patient's talk. The companion's communication may influence not only the patient's but also the physician's communication. The patient's expectation of the companion's role during the visit and the companion's intention regarding their role were generally related to one another, and had positive associations with the companion's actual behavior during the visit. Nevertheless, companions often anticipated playing a more direct communication role during the visit, including the provision of information and asking of questions, than patients expected of them. Further investigation is needed to explore the communication dynamics in triads and dyads, and its relation to patient outcomes. PMID- 15748679 TI - Quality of life and coping strategies among lung transplant candidates and their family caregivers. AB - Although numerous studies have examined coping strategies and quality of life (QOL) among patients with chronic diseases and their family caregivers, no studies have examined the reciprocal effects of patient and caregiver coping strategies on their dyad partner's QOL. Because most people who cope with stressful health experiences do so within the context of interpersonal relationships, it is important to understand the ways in which the two partners' coping strategies may reciprocally affect each other's QOL. Adult lung transplant candidates and their caregivers (N=114 pairs) participated in semi-structured interviews that included measures of QOL and coping with patients' health-related problems. Multivariate, canonical correlation analyses were performed to examine unique patterns of associations between coping and QOL in patient-caregiver dyads. Better patient QOL, across multiple domains, was associated with better caregiver QOL. Multiple elements of patients' coping, including greater use of active coping and emotionally oriented coping were related to generally poorer patient QOL in psychosocial and physical domains. Similarly, caregivers who used more emotionally oriented coping had poorer QOL. There was no statistically reliable relationship between either (a) patient and caregiver use of coping strategies, or (b) caregiver coping and patient QOL. However, patients' coping strategies were important correlates of caregivers' QOL. These findings belie common clinical beliefs that family members' coping responses to patients' health are likely to affect patient well-being. Instead, patients' coping and QOL may be critical for understanding caregiver well-being, especially in the current era in which caregivers are assuming increased responsibility for providing patient care. PMID- 15748681 TI - Parents' perceptions of pediatric day surgery risks: unforeseeable complications, or avoidable mistakes? AB - A decline in people's trust of the US health care system has been documented. But do parents of pediatric patients have specific safety worries? If so, what are they? And what cultural factors inform them? To help answer these questions, in San Diego 35 English- and Spanish-speaking parents' (or guardians') self-reports of perceptions of their child's risk for experiencing a medical error during day surgery were collected using open-ended rapid assessment interviews, and analyzed using qualitative content analysis methods. (The innovative data collection and analysis protocol is described so as to be easily replicable in other settings for other types of highly focused quality improvement initiatives that rely on the inclusion of parent or patient perspectives and entail limited time and resources). Twelve key themes were identified. Anthropological discourse analysis techniques (modified for rapid use with rapidly collected interview data) were then applied to the transcripts in order to generate ideas regarding cultural factors underlying the themes. Discursive patterns of risk rationalization or refutation related to cultural expectations for parenting, children's bodies, and health care consumerism were found. Neither the (careful, loving, good) parent nor the (wisely chosen or well-reputed) care team was to blame for medical errors. Errors were cast as the inevitable results of latent, unanticipatable vulnerabilities inherent in the (defenseless) children undergoing surgery. The anxiety-generating potential of the surgical consent process also was noted. The implications of the findings for action as well as for theories of risk and vulnerability and of childhood and parenthood are discussed. PMID- 15748680 TI - Participatory action research: considerations for ethical review. AB - This paper addresses the distinctive nature of participatory action research (PAR) in relation to ethical review requirements. As a framework for conducting research and reducing health disparities, PAR is gaining increased attention in community and public health research. As a result, PAR researchers and members of Research Ethics Boards could benefit from an increased understanding of the array of ethical concerns that can arise. We discuss these concerns in light of commonly held ethical requirements for clinical research (social or scientific value, scientific validity, fair subject/participant selection, favourable risk benefit ratio, independent review, informed consent, and respect for potential and enrolled participants) and refer to guidelines specifically developed for participatory research in health promotion. We draw from our community-based experiences in mental health promotion research with immigrant and culturally diverse youth to illustrate the ethical advantages and challenges of applying a PAR approach. We conclude with process suggestions for Research Ethics Boards. PMID- 15748682 TI - The impact of advertising on nicotine replacement therapy demand. AB - While much is known about the economic determinants of tobacco use, very little is known about the economic determinants of nicotine replacement therapy (NRT) use. This paper is the first econometric study to examine the impact of advertising on NRT demand. Pooled cross-sectional time-series scanner-based data for 50 major metropolitan markets in the USA covering the period between the second quarter of 1996 and the second quarter of 2002 are used in the analysis. Fixed-effects modeling is employed to estimate the NRT demand equation. The estimates indicate that increased advertising of Nicoderm CQ transdermal patches and Nicotrol transdermal patches increases per-capita sales of established Nicoderm CQ and Nicotrol products, respectively. However, increased advertising of Nicorette polacrilex (gum) was found not to significantly increase sales of established Nicorette products. Moreover, decreases in the price of NRT and increases in the price of cigarettes were found to increase per-capita sales of NRT products. Given the documented efficacy of NRT, measures to increase peoples' awareness of NRT products through advertising, measures to decrease the price of NRT, and measures to increase the price of cigarettes would be effective means to increase the use of NRT, likely leading to decreased cigarette smoking and reductions in the future public health burden caused by tobacco use. PMID- 15748683 TI - Governance in gridlock in the Russian health system; the case of Sverdlovsk oblast. AB - Epidemiological, demographic and environmental crises, transition to a new political regime and exceptionally severe economic crises were powerful stimuli to health sector reform in Russia. The Russian Federation responded by introducing medical insurance whilst decentralising public administration. Yet despite intense contextual pressures to do so and a new policy climate, Russian hospitals found it difficult to reprofile services and reallocate their resources. A case study analysing governance structures in Sverdlovsk oblast reveals that medical insurance created incentives to reduce costs by reducing bed days, but if hospitals did so they would lose money under the formulae through which decentralised local government still allocated around three-quarters of hospital income. If instead hospitals tried to increase budgetary income by increasing numbers of bed-days, the insurance system would penalise them. This specific form of policy mess can be called 'governance in gridlock'. The juxtaposition of two overlapping but incompatible sets of governance structures practically immobilised official hospital management systems. It is as one-sided to blame residues of the Soviet system for this gridlock as it is to blame the medical insurance system. Gridlock resulted from the interaction of the two, a problem to which all health system reform is potentially vulnerable. PMID- 15748684 TI - Credit program outcomes: coping capacity and nutritional status in the food insecure context of Ethiopia. AB - This paper presents findings of a survey that was primarily intended as (1) an assessment of coping capacity in drought and food insecure conditions and (2) a microfinance program outcome study. A three group cross-sectional survey of 819 households was conducted in May 2003 in two predominantly rural sites in Ethiopia. Established clients of the WISDOM Microfinance Institution were compared with similar incoming clients and community controls. No overall pattern of enhanced prevalence of coping mechanisms was observed in any participant group, suggesting that participation in the lending program did not affect coping capacity at the household level. No significant differences in mean mid-upper arm circumference or prevalence of acute malnutrition were found in males or females when the total sample was assessed. In the primary survey site, Sodo, female clients and their children had significantly better nutritional status than other comparison groups: the odds of malnourishment in female community controls compared to established female clients was 3.2 (95% CI: 1.1-9.8) and the odds of acute malnutrition in children 6-59 months of age were 1.6 times greater in children of both male clients and community controls (95% CI: .78-3.32). Household food security among female client households in Sodo was significantly better than in other comparison groups according to a variety of indicators. As compared to female clients, male clients and community controls, respectively, were 1.94 (95% CI: 1.05-3.66) and 2.08 (95% CI: 1.10-4.00) times more likely to have received food aid during the past year. Findings of the present study suggest that microfinance programs may have an important impact on nutritional status and well-being of female clients and their families. That female clients were significantly less likely to be food aid recipients suggests that microfinance programs may be successful in reducing vulnerability to prolonged drought and food insecurity. PMID- 15748685 TI - Beyond intention: do specific plans increase health behaviours in patients in primary care? A study of fruit and vegetable consumption. AB - Increasing evidence suggests that implementation intentions are effective in moving people towards achieving health behaviour goals. However, the type of health behaviours for which they work best is unclear. Furthermore, implementation intentions appear to be less effective when studied in clinical rather than student populations. This prospective study tested implementation intentions with a complex, repeated health behaviour in a patient sample. A total of 120 cardiac patients in the UK were asked to increase their daily fruit and vegetable consumption by two portions and to maintain this over 3 months. Participants were randomly assigned to three groups (control, Theory of Planned Behaviour (TPB) questionnaire, TPB questionnaire+implementation intention) and telephoned at 7, 28 and 90 days follow-up to record daily consumption (24-h recall measure); 94 participants completed the study. Daily fruit and vegetable consumption increased from 2.88 portions (SD=1.67) at recruitment to 4.28 portions (SD=2.25) at 90 days. A 4x3 (time by group) mixed design ANCOVA was computed with daily fruit and vegetable consumption at recruitment entered as a covariate. This revealed a significant time effect (F (3, 270)=29.79, p<0.001) (eta2=0.25) but non-significant group (F (2, 90)=0.32, p=0.73) (eta2=0.07) and time by group effects (F (6, 270)=0.48, p=0.82) (eta2=0.01). There was also a significant main effect of the covariate (F (1, 90)=48.51, p<0.001) (eta2=0.35) and a significant time by covariate effect (F (3, 270)=12.14, p<0.001) (eta2=0.12). Substantial increases in fruit and vegetable consumption were achieved particularly by participants who were eating low levels at recruitment. Consumption was not improved by implementation intentions. These findings are discussed in the context of the targeted health behaviour and sample. PMID- 15748686 TI - A pharmacist model of perceived responsibility for drug therapy outcomes. AB - Pharmacists in community and ambulatory care settings are in a unique position to reduce drug-related morbidity and to optimize patient outcomes by identifying, resolving, and preventing drug therapy problems. This particular approach to pharmacy practice expands traditional pharmacist responsibilities of dispensing pharmaceuticals and providing drug information to optimizing patients' drug therapy outcomes. However, pharmacists in general, and community pharmacists in particular, have yet to incorporate this expanded professional role into daily practice. The objective of this study was to examine the validity of a pharmacist model of perceived responsibility for drug therapy outcomes based on the triangle model of responsibility. A survey instrument was tested among community and ambulatory care pharmacists in Florida, USA. The survey instrument contained the following pharmacist-related constructs from the model: clarity of standards, personal control, professional duty, and perceived responsibility for drug therapy outcomes. The model was examined by testing hypothesized relationships between the model constructs and pharmacists' reports of providing pharmaceutical care. The survey response rate was 40.9% (525/1283). All of the study measures exhibited Cronbach alpha values greater than .70. A measurement model was tested using confirmatory factor analysis. The chi2/df ratio (3.02), CFI (.95), and residual (.051) indicated a good fit of the item data to the constructs. According to path analysis, clarity of standards, personal control, and professional duty were significantly related to perceived responsibility for drug therapy outcomes, which in turn, was significantly related to pharmaceutical care provision. Perceived responsibility for drug therapy outcomes acted as a mediator of the effects of clarity of standards, personal control, and professional duty on pharmaceutical care provision. These findings have implications for pharmacy practice and research. PMID- 15748687 TI - Cloneless genomic DNA analysis: an efficient and simple methods for de novo genomic sequencing projects and gap filling. AB - The utility of using genomic DNA directly in agarose, i.e. cloneless libraries, in place of large clone libraries, radiation hybrid panels, or chromosome dissection was demonstrated. The advantage of the cloneless library approach is that, in principle, a targeted genomic resource can be developed rapidly for any genomic region using any genomic DNA sample. Here, a human chromosome 20 Not I fragment library was generated by slicing a pulsed field gel lane containing fractionating Not I cleaved DNA from a monosomic hybrid cell line into 2 mm pieces. A reliable PCR method using agarose embedded DNA was developed. InterAlu PCR generated unique patterns of products from adjacent slices (e.g. fractions). Further, the specificity of the interAlu products was demonstrated by FISH analysis and in other hybridization experiments to arrayed interAlu products. STS content mapping was used to order the fractions and also demonstrate the unique content of the library fractions. PMID- 15748689 TI - Micropillar array chip for integrated white blood cell isolation and PCR. AB - We report the fabrication of silicon chips containing a row of 667 pillars, 10 by 20 microm in cross-section, etched to a depth of 80 microm with adjacent pillars being separated by 3.5 microm. The chips were used to separate white blood cells from whole blood in less than 2 min and for subsequent PCR of a genomic target (eNOS). Chip fluid dynamics were validated experimentally using CoventorWare microfluidic simulation software. The amplicon concentrations were determined using microchip capillary electrophoresis and were >40% of that observed in conventional PCR tubes for chips with and without pillars. Reproducible on-chip PCR was achieved using white blood cell preparations isolated from whole human blood pumped through the chip. PMID- 15748690 TI - Effect of low oxygen concentrations on growth and alpha-amylase production of Aspergillus oryzae in model solid-state fermentation systems. AB - Oxygen transfer in the fungal mat is a major concern in solid-state fermentation (SSF). Oxygen supply into the mycelial layers is hampered by diffusion limitation. For aerobic fungi, like Aspergillus oryzae, this oxygen depletion can be a severely limiting factor for growth and metabolite production. This paper describes the effects of a low oxygen concentration on growth at the levels of individual hyphae, colonies and overcultures, and on alpha-amylase production in overcultures. PDA medium was used to study the effect of a low oxygen concentration on hyphal elongation rate and branching frequency of hyphae, and radial extension rate of colonies of A. oryzae. We found similar saturation constants (K(O2)) of 0.1% (v/v in the gas phase) for oxygen concentration described with Monod kinetics, for branching frequency of hyphae and colony extension rate. When A. oryzae was grown as an over-culture on wheat-flour model substrate at 0.25% (v/v) oxygen concentration, the reduction in growth was more pronounced than as individual hyphae and a colony on PDA medium. Experimental results also showed that the specific alpha-amylase production rate under the condition of 0.25% (v/v) oxygen was reduced. Because the value of K(O2) is relatively low, it is reasonable to simplify the kinetics of growth of A. oryzae to zero-order kinetics in coupled diffusion/reaction models. PMID- 15748688 TI - A human biotin acceptor domain allows site-specific conjugation of an enzyme to an antibody-avidin fusion protein for targeted drug delivery. AB - We have previously constructed an antibody-avidin (Av) fusion protein, anti transferrin receptor (TfR) IgG3-Av, which can deliver biotinylated molecules to cells expressing the TfR. We now describe the use of the fusion protein for antibody-directed enzyme prodrug therapy (ADEPT). The 67 amino acid carboxyl terminal domain (P67) of human propionyl-CoA carboxylase alpha subunit can be metabolically biotinylated at a fixed lysine residue. We genetically fused P67 to the carboxyl terminus of the yeast enzyme FCU1, a derivative of cytosine deaminase that can convert the non-toxic prodrug 5-fluorocytosine to the cytotoxic agent 5-fluorouracil. When produced in Escherichia coli cells overexpressing a biotin protein ligase, the FCU1-P67 fusion protein was efficiently mono-biotinylated. In the presence of 5-fluorocytosine, the biotinylated fusion protein conjugated to anti-rat TfR IgG3-Av efficiently killed rat Y3-Ag1.2.3 myeloma cells in vitro, while the same protein conjugated to an irrelevant (anti-dansyl) antibody fused to Av showed no cytotoxic effect. Efficient tumor cell killing was also observed when E. coli purine nucleoside phosphorylase was similarly targeted to the tumor cells in the presence of the prodrug 2-fluoro-2'-deoxyadenosine. These results suggest that when combined with P67-based biotinylation, anti-TfR IgG3-Av could serve as a universal delivery vector for targeted chemotherapy of cancer. PMID- 15748691 TI - Fine structural and cytochemical mapping of enamel organ during the enameloid formation stages in gars, Lepisosteus oculatus, Actinopterygii. AB - During cap enameloid formation in gars (Lepisosteus oculatus), the dental epithelial cells that constitute the enamel organ were observed by means of transmission electron microscopy and enzyme cytochemistry to detect the hydrolytic enzyme activities, alkaline phosphatase (ALPase), acid phosphatase (ACPase), calcium-dependent adenosine triphosphatase (Ca-ATPase) and potassium dependent p-nitrophenylphosphatase (K-NPPase) (sodium, potassium-activated adenoshine triphosphatase (Na-K-ATPase)). The enameloid formation process in gars was divided into three stages: matrix formation, mineralisation and maturation. The enamel organ consisted of the outer dental epithelial (ODE) cells, stellate reticulum (SR), stratum intermedium (SI) and the inner dental epithelial (IDE) cells during the whole of the cap enameloid formation stages. During the matrix formation stage, many cisternae of rough endoplasmic reticulum and widely distributed Golgi apparatus, in which the procollagen granules containing cross striations were often found, were remarkable elements in the IDE cells. During the stage of mineralisation, the IDE cells were tall columnar, and infoldings of distal plasma membrane of the IDE cells became marked. The most developed Golgi apparatus was visible at this stage, and large secretory granules containing fine granular or tubular materials were found in the distal cytoplasm that was close to the infoldings of the distal end. Many lysosomes that were ACPase positive were seen near the Golgi apparatus and in the distal cytoplasm of the IDE cells. ACPase positive granules often contained the cross-striation structure resembling procollagen, suggesting that the procollagen is degenerated in the IDE cells. During the maturation stage, the distal infoldings became unclear, and there were no large granules containing tubular materials, but many ACPase positive lysosomes were still present in the IDE cells. Non-specific ALPase was detected at the plasma membrane of the IDE cells at the mineralisation and maturation stages. K-NPPase was markedly detected at the plasma membrane of the IDE cells at the maturation stage. These results demonstrate that the IDE cells might be mainly involved in the removal of degenerated organic matrix from enameloid during the later formation stages. Strong Ca-ATPase activity was observed at the entire plasma membrane of the stratum intermedium cells, and there was slightly weak activity at the plasma membrane of the IDE cells during the mineralisation and maturation stages, implying that these cells are related to the active Ca transport to the maturing enameloid. It is likely that although the structure of the enamel organ is different, the function, especially at the mineralisation and maturation stages, is similar to other actinopterygians having well-mineralized cap enameloid. PMID- 15748692 TI - GD3 synthase gene found expressed in dental epithelium and shown to regulate cell proliferation. AB - GD3 synthase is one of the key enzymes involved with ganglioside synthesis, and its activity regulates the main profile of ganglioside expression. We analyzed the expression of the GD3 synthase gene in laser-dissected teeth germs using RT PCR. The GD3 synthase gene was found expressed in brain, thymus, and tooth germ tissues, however, not in liver or skin specimens. Further, it was highly expressed during the early stage of tooth germ development (embryonic day 14.5), especially in dental epithelia, which gradually reduced in the molar site until postnatal day 7, whereas it was not in dental mesenchyme tissues. In addition, dental epithelial cells transiently transfected with the GD3 synthase gene showed enhanced proliferation. These results indicate that the GD3 synthase gene may be involved in early tooth development, particularly in the proliferation of dental epithelium. PMID- 15748693 TI - Origin and developmental fate of vestigial tooth primordia in the upper diastema of the field vole (Microtus agrestis, Rodentia). AB - OBJECTIVE: Odontogenesis in voles is a convenient model to test hypotheses on tooth development generated from investigations in the mouse. Similar to other rodents, the functional dentition of the vole includes a toothless diastema. At its mesial end, a vestigial tooth bud has been found in the upper jaw of vole embryos. The aim of this study was to analyse the developmental dynamics of vestigial tooth structures in the upper diastema of the field vole and to compare it with the situation in the mouse. DESIGN: The development of odontogenic structures in the upper diastema of the field vole was investigated using serial histological sections and three-dimensional (3D) computer-aided reconstruction. RESULTS: A transient continuous dental lamina in the upper diastema of the field vole extended mesially to the first molar primordium, but was not continuous with the dental lamina in the incisor region. At its mesial limit, a large vestigial tooth primordium was regularly present. A further distinct vestigial bud was located mesially to the first molar primordium. The segmentation of the dental lamina suggested a potential to give rise to further vestiges in the upper diastema of the vole. CONCLUSIONS: In the prospective diastema of the vole exists as in the mouse a continuous dental lamina. Beside the prominent vestigial tooth bud in the mesial diastema, a further large bud was transiently located in front of the molars. The incorporation of dental epithelium into the first upper molar (M(1)) primordium in the vole differs from that in the mouse. PMID- 15748694 TI - Ultrasound to stimulate early bone formation in a distraction gap: a double blind randomised clinical pilot trial in the edentulous mandible. AB - OBJECTIVE: In a double blind randomised clinical pilot trial, it was investigated whether low intensity pulsed ultrasound therapy stimulates early bone formation in a distraction gap created in a severely resorbed mandible. DESIGN: Eight patients underwent a mandibular vertical distraction over an average distance of 6.6+/-1.1mm. Ultrasound self-therapy or placebo therapy was started on the first day of distraction and continued daily until the implants were inserted. After 31+/-3.8 days of consolidation, the distraction device was removed, a transmandibular biopsy was taken, and two endosseous implants were inserted. RESULTS: All patients complied well with ultrasound therapy. During an average of 30.1+/-4.1 months follow-up, no complications did occur. Microradiographic examination of the biopsies revealed a comparable mean area of mineralised tissue in the distraction gap of 1.9+/-1.7mm(2) in the ultrasound treatment group and 1.9+/-1.3mm(2) in the placebo treatment group. Histological examination indicated that active woven bone was present within the distraction gap just adjacent to the osteotomy plane, with no apparent differences between the treatment groups. The lamellar bone formation outside the distraction gap appeared to have started as well. CONCLUSION: During a 31-day consolidation period, ultrasound treatment does not appear to stimulate bone formation in the severely resorbed vertical distracted mandible and it seems that this period is too short to evaluate properly if there is an effect. Therefore, a longer consolidation period has to be studied. PMID- 15748695 TI - Effect of intermittent PTH administration in the periodontitis-associated bone loss in ovariectomized rats. AB - OBJECTIVE: Parathyroid hormone intermittent administration has been considered to treat bone mass decrease in osteoporotic individuals. The present study evaluates whether PTH can affect alveolar bone loss in ovariectomized rats, since estrogen deficiency has been proposed as a risk factor for periodontal disease. DESIGN AND METHODS: Thirty female rats were set in groups: ovariectomized (Ovx) and Sham operated. Ovx were divided in two groups: Ovx-PTH (1-34) treated and Ovx, which received vehicle. After 1 week, cotton ligature was placed around one lower first molar of all animals to induce periodontal disease. Ovx treated received PTH doses of 40 microg/kg, three times a week for 30 days. After that, the animals were sacrificed, the mandibles extracted, X-rayed and samples prepared for histological evaluation. Histomorphometry was performed using image analyzer software. Scanning electron microscopy (SEM) of the tibias was also performed in all animals to evaluate possible changes in bone structure caused by the estrogen deficiency. Optical densities of the radiographs were measured by aluminum step wedge equivalent thickness. RESULTS: Histomorphomery indicated the anabolic PTH effect in ovariectomized rats with significant inhibition of periodontitis manifestation (p<0.05) thus neutralizing the periodontitis inductor effects. The photo densitometry showed a lower mandibular optical density in the ovariectomized group that did not receive PTH (p<0.05). SEM image confirmed the early effect of estrogen deficiency in osseous tissue and PTH anabolic effect. CONCLUSION: PTH systemic intermittent administration was able to reduce alveolar bone loss in ovariectomized rats, despite the presence of a periodontal disease inductor and estrogen deficiency. PMID- 15748696 TI - The differentiation profile of the epithelium of the human lip. AB - The aim of this study was to analyse the immunohistochemical differentiation profile of the stratified squamous epithelium of the adult human lip. Full thickness lower lips taken from 31 cadavers were analysed. Sections were stained with haematoxylin and eosin, periodic acid-Schiff (PAS), cytokeratins (CK), loricrin, involucrin, profilaggrin and filaggrin. The stratified squamous epithelium covering the lip could be divided into: (i) appendage-bearing, orthokeratinised epidermis; (ii) orthokeratinised vermilion which had a more prominent rete pattern than the epidermis; (iii) parakeratinised, PAS-positive intermediate zone; and (iv) non- or parakeratinised labial mucosal epithelium. Epithelial thickness increased gradually from the skin to the mucosal aspect. The CK pattern changed across the intermediate zone, with gradual loss of CK 1 and 10 from the skin, and CK 4, 13 and 19 from the mucosal, aspect. CK 5 and 14 were consistently expressed basally, and variably expressed suprabasally. Apart from labelling Merkel cells, CK 8, 18 and 20 were negative. Involucrin, which was present at all sites, was restricted to the stratum granulosum in skin, but extended into the stratum spinosum, and gradually into parabasal keratinocytes, across the vermilion and mucosa. Loricrin, profilaggrin and filaggrin were present in the stratum granulosum of orthokeratinised sites, but expression was abruptly lost at the junction between the vermilion and the intermediate zone. In conclusion, the phenotype of the stratified squamous epithelium covering the lip changes at, or across, the intermediate zone of the adult vermilion. It is possible that changes in the composition of the stratified squamous epithelium affect the colour of the vermilion. PMID- 15748697 TI - Distinct fibro-vascular arrangements in the periodontal ligament of the horse. AB - OBJECTIVE: The periodontal ligament (PDL) of the hypsodont cheek teeth of the horse meets specific requirements of tooth support: (1) to fix the tooth; (2) to withstand pressure and tension during the laterolateral mastication movement; (3) to restore the original tooth position. These requirements are assumed to be met by the periodontal collagen fiber apparatus and by the vascular system. Distinct fibro-vascular arrangements are described here in order to reveal the structural prerequisites for these distinct functions. DESIGN: Specimens from nine horses were processed for (1) collagen labeling with fluorescein, (2) oxytalan fiber staining, (3) immunostaining of smooth muscle actin, and (4) TEM examinations. RESULTS: Blood vessels were found to be integrated in the PDL in three distinct ways: CONCLUSIONS: The sheaths formed by veil cells (type-I arrangement) isolate distinct groups of blood vessels from deformations of the surrounding tissue. In contrast, the distinct fibro-vascular arrangements of anchored blood vessels (type-II arrangement) and ballooned venules (type-III arrangement) are assumed to accept the forces generated by masticatory tooth movements. The anchored blood vessels sustain tractive forces by means of what is called a lateral compression mechanism, whereas the ballooned venules act as a hemodynamic cushion. PMID- 15748698 TI - Mechanical deformation inhibits IL-10 and stimulates IL-12 production by mouse calvarial osteoblasts in vitro. AB - The skeleton is continuously remodelled throughout life, a process that is orchestrated by cells of the osteoblast lineage. Remodelling involves a complex network of cell-cell signalling involving systemic hormones, locally produced cytokines, growth factors and the mechanical environment of the cells. Here, we report on the effect of mechanically-induced strain on the synthesis by mouse calvarial osteoblasts in monolayer culture of IL-10 and IL-12, two cytokines that inhibit osteoclast formation in bone marrow cultures; IL-10 also suppresses osteoblast differentiation suggesting a role for both cytokines in bone physiology. A tensile strain was applied to the cells intermittently for 6s, every 90s, for 2-96h. After 2-h culture, supernatants from deformed cells contained significantly less IL-10 than control cultures. In contrast, mechanical deformation had a stimulatory effect on IL-12 synthesis; however, by 48h both had returned to control levels. These data suggest that IL-10 and IL-12 can be added to the growing list of mechanical stress-responsive genes. The down-regulation of IL-10 and stimulation of IL-12 further suggests that the initial response of the cells to mechanical deformation was an osteogenic one. PMID- 15748699 TI - Inhibitory effects on human eosinophil chemotaxis in vitro by BAY 41-2272, an activator of nitric oxide-independent site of soluble guanylate cyclase. AB - This study was designed to investigate the effects of the 5-cyclopropyl-2-[1-(2 fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-pyrimidin-4-ylamine (BAY 41-2272) on formyl-methionyl-leucyl-phenylalanine (fMLP; 10(-7)M)-induced human eosinophil chemotaxis, cyclic guanosine-3',5'-monophosphate (cGMP) and cyclic adenosine 3',5'-monophosphate (cAMP) levels. Human eosinophils were pretreated or not with 3-isobutyl-l-methyl-xanthine (IBMX; 500microM), and then exposed to BAY 41-2272 (0.1-10.0microM) for either short (10min) or prolonged (90min) time periods. Exposition of eosinophils with BAY 41-2272 for either 10min or 90min markedly inhibited the eosinophil chemotaxis, independently of IBMX pretreatment. Inhibition of fMLP-induced eosinophil chemotaxis by BAY 41-2272 (in absence of prior treatment with IBMX) was about of the same irrespective if cells were exposed for 10min or 90min with this compound. In IBMX-pretreated eosinophils, the inhibition of fMLP-induced chemotaxis by BAY 41-2272 in the 10-min exposure protocols was even higher in comparison with the 90-min protocols. Incubation of IBMX-treated eosinophils for 90min with BAY 41-2272 resulted in 2.0-2.5 times higher levels of cGMP and cAMP compared with the 10-min protocols. The BAY 41 2272-induced cGMP increases were abolished by pre-incubation of eosinophils with the soluble guanylate cyclase inhibitor 1H-[1,2,4]-oxidiazolo[4,3-a] quinoxalin-1 one (ODQ). No eosinophil toxicity was observed in any experimental condition, according to 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. Our findings show that inhibitory effects of fMLP-induced human eosinophil chemotaxis by BAY 41-2272 at short-term or prolonged exposition time are accompanied by significant elevations of cGMP and cAMP, but we could not detect a clear correlation between chemotaxis inhibition and elevation of cyclic nucleotide levels. PMID- 15748700 TI - The role of adenosine A2A and A2B receptors in the regulation of TNF-alpha production by human monocytes. AB - Adenosine is an endogenous nucleoside that regulates many physiological processes through the activation of its four receptors: A(1), A(2A), A(2B) and A(3). Previous studies have identified the involvement of A(2) receptors in the inhibitory activity of adenosine analogues on tumor necrosis factor-alpha (TNF alpha) production by lipopolysaccharide (LPS) activated monocytes, but the relative contributions of A(2A) versus A(2B) receptors have not been determined in human primary monocytes. Nor has the role of A(1) and A(3) been clearly identified in the system. The lack of such information impacts on the selection of adenosine receptor agonists for disease intervention. Using LPS-stimulated human primary monocytes, we found that the adenosine receptor agonist, 5'-N ethylcarboxamidoadenosine (NECA) or the A(2A) receptor agonist, 4-[2-[[6-amino-9 (N-ethyl-b-d-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride (CGS21680) produced a concentration-dependent inhibition of TNF-alpha production, with IC(50)s of 58.4nM (32.7-104.5nM, 95% confidence interval) and 49.2nM (22.7-105.9nM, 95% confidence interval), respectively. The selective A(2A) receptor blocker, 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3 a][1,3,5]triazin-5-ylaminso]ethyl)phenol (ZM241385, 30nM), antagonized the effects of NECA and CGS21680 (pK(B) estimates were 8.7+/-0.1 and 8.9+/-0.1, respectively), while the selective A(2B) antagonist, N-(4-cyano-phenyl)-2-[4-(2,6 dioxo-1,3-dipropyl-2,3,4,5,6,7-hexahydro-1H-purin-8-yl)-phenoxy]-acetamide (MRS1754, 100nM), failed to antagonize the effects of either agonist. Furthermore, neither the A(1) receptor agonist, 2-chloro-N(6) cyclopentyladenosine (CCPA) nor the A(3) receptor agonist, 1-[2-chloro-6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-1-deoxy-N-methyl-b-d-ribofuranuronamide (2-Cl-IB-MECA) showed significant inhibitory activity at concentrations that effectively bind to their respective receptors. We conclude that A(2A) receptor activation is predominantly responsible for the inhibitory effects of adenosine receptor agonists on TNF-alpha production from LPS-stimulated monocytes. PMID- 15748701 TI - Sodium nitroprusside-induced osteoblast apoptosis is mediated by long chain ceramide and is decreased by raloxifene. AB - Release of high levels of nitric oxide (NO) is associated with osteoblastic cell death. The mechanisms of NO-induced cytotoxicity are not well documented and it is presently not known if estrogenic compounds prevent this effect. We studied the role of ceramides in cell death induced by the NO donor sodium nitroprusside (SNP) and we tested the possibility that 17beta-estradiol, the anti-estrogen ICI 182.780 and two selective estrogen receptor modulators raloxifene and tamoxifen modify osteoblastic cell apoptosis. SNP dose-dependently decreased MC3T3-E1 osteoblast viability, increased NO production in the culture media and enhanced the release of intracellular ceramides C22 and C24. Cell death induced by SNP was partially inhibited when MC3T3-E1 cells were pretreated with raloxifene and tamoxifen but was not modified when the cells were pretreated with 17beta estradiol or ICI 182.780. Cell death induced by SNP resulted from apoptosis as demonstrated by Annexin-V and propidium iodide labeling and a reduction of SNP induced MC3T3-E1 apoptosis was confirmed in the presence of raloxifene and tamoxifen. SNP induction of C22 and C24 production was inhibited by a pretreatment with raloxifene but not with 17beta-estradiol. Moreover, the synthetic ceramide C24 (0.75 and 1microM) decreased MC3T3-E1 cell viability and osteoblast cell death induced by C24 was partially decreased by raloxifene and to a lesser extent by 17beta-estradiol. These data demonstrate that SNP-induced cell death is mediated by the long chain ceramides C22 and C24 and that raloxifene protected osteoblast from apoptosis induced by SNP, an effect that might be relevant to its pharmacological properties on bone remodeling. PMID- 15748702 TI - Antioxidant, prooxidant and cytotoxic activity of hydroxylated resveratrol analogues: structure-activity relationship. AB - Resveratrol (trans-3,4',5-trihydroxystilbene), a naturally occurring hydroxystilbene, is considered an essential antioxidative constituent of red wine possessing chemopreventive properties. However, resveratrol and even more its metabolite piceatannol were reported to have also cytostatic activities. In order to find out whether this is related to antioxidative properties of those compounds, we synthesized five other polyhydroxylated resveratrol analogues and studied structure-activity relationships between pro-/antioxidant properties and cytotoxicity. Radical scavenging experiments with O(2)(*-) (5,5-dimethyl-1 pyrroline-N-oxide/electron spin resonance (DMPO/ESR)) and 2,2-diphenyl-1 picrylhydrazyl (DPPH) (photometry) revealed that 3,3',4',5-tetrahydroxystilbene (IC(50): 2.69microM; k(9): 443000M(-1)s(-1)), 3,4,4',5-tetrahydroxystilbene (IC(50): 41.5microM; k(9): 882000M(-1)s(-1)) and 3,3',4,4',5,5' hexahydroxystilbene (IC(50): 5.02microM), exerted a more than 6600-fold higher antiradical activity than resveratrol and its two other analogues. Furthermore, in HL-60 leukemic cells hydroxystilbenes with ortho-hydroxyl groups exhibited a more than three-fold higher cytostatic activity compared to hydroxystilbenes with other substitution patterns. Oxidation of ortho-hydroxystilbenes in a microsomal model system resulted in the existence of ortho-semiquinones, which were observed by ESR spectroscopy. Further experiments revealed that these intermediates undergo redox-cycling thereby consuming additional oxygen and forming cytotoxic oxygen radicals. In contrast to compounds with other substitution patterns hydroxystilbenes with one or two resorcinol groups (compounds 1 and 3) did not show an additional oxygen consumption or semiquinone formation. These findings suggest that the increased cytotoxicity of ortho-hydroxystilbenes is related to the presence of ortho-semiquinones formed during metabolism or autoxidation. PMID- 15748703 TI - Mitochondrial-dependent, reactive oxygen species-independent apoptosis by myricetin: roles of protein kinase C, cytochrome c, and caspase cascade. AB - Abrogation of mitochondrial permeability and induction of reactive oxygen species (ROS) production have been observed in chemical-induced apoptosis; however, the relationship between the mitochondria and intracellular ROS levels in apoptosis is still unclear. In the present study, myricetin (ME) but not its respective glycoside, myricitrin (MI; myricetin-3-O-rhamnose) reduced the viability of human leukemia HL-60 cells via apoptosis, characterized by the occurrence of DNA ladders and hypodiploid cells. Results of Western blotting and caspase activity assays showed that activation of caspases 3 and 9 but not caspases 1, 6 or 8 with cleavage of PARP and D4-GDI proteins is involved in ME-induced apoptosis. A reduction in mitochondrial functions characterized by a decrease in the Bcl-2/Bax protein ratio and translocation of cytochrome c (cyt c) from the mitochondria to the cytosol in accordance with a decrease in mitochondrial membrane potential were observed in ME-treated HL-60 cells. No significant induction of intracellular ROS levels by ME was observed by the DCHF-DA assay, DPPH assay or plasmid digestion assay, and antioxidants including N-acetyl-cysteine (NAC), catalase (CAT), superoxide dismutase (SOD), and tiron (TIR) showed no protective effects on ME-induced apoptosis. A PKC activator, 12-O-tetradecaoylphorbol-13 acetate (TPA) significantly attenuated ME-induced apoptosis via preventing cytochrome c release to the cytosol and maintaining the mitochondrial membrane potential by inhibiting the decrease in the Bcl-2/Bax protein ratio; these effects were blocked by protein kinase C (PKC) inhibitors including GF-109203X, H7, and staurosporin. Removing mitochondria by ethidium bromide (EtBr) treatment reduced the apoptotic effect of ME. Results of SAR studies showed that the presence of OH at C3', C4', and C5' is important for the apoptosis-inducing activities of ME, and that ME induces apoptosis in another leukemia cell line, Jurkat cells, but not in primary human polymorphonuclear (PMN) cells or in murine peritoneal macrophages (PMs). The results of the present study suggest that apoptosis induced by ME occurs through a novel mitochondrion-dependent, ROS independent pathway; TPA protects cells from ME-induced apoptosis via PKC activation which prevents the occurrence of mitochondrial destruction during apoptosis. PMID- 15748704 TI - Suppression of Ca2+ influx by unfractionated heparin in non-excitable intact cells via multiple mechanisms. AB - Effect of unfractionated heparin (UFH), described as a cell-impermeant IP3 receptor antagonist, was studied on the capacitive Ca(2+) entry in non permeabilized, intact cells, measuring the intracellular Ca(2+) levels using fluorescence microplate technique. Ca(2+) influx induced via Ca(2+) mobilization by histamine in Hela cells or evoked by store depletion with thapsigargin in RBL 2H3 cells was dose-dependently suppressed by UFH added either before or after the stimuli. UFH also prevented the spontaneous Ba(2+) entry indicating that the non capacitive Ca(2+) channels may also be affected. In addition, UFH caused a significant and dose-dependent delay in Ca(2+), and other bivalent cation inflow after treatment of the cells with Triton X-100, but it did not diminish the amount of these cations indicating that UFH did not act simply as a cation chelator, but modulated the capacitive Ca(2+) entry possibly via store operated Ca(2+) channels (SOCCs). Inhibitory activities of UFH and 2-aminoethyl diphenyl borate on the capacitive Ca(2+) influx was found reversible, but the time courses of their actions were dissimilar suggesting distinct modes of action. It was also demonstrated using a fluorescence potentiometric dye that UFH had a considerable hyperpolarizing effect and could alter the changes of membrane potential during Ca(2+) influx after store depletion by thapsigargin. We presume that the hyperpolarizing property of this agent might contribute to the suppression of Ca(2+) influx. We concluded that UFH can negatively modulate SOCCs and also other non-capacitive Ca(2+) channels and these activities might also account for its multiple biological effects. PMID- 15748705 TI - Involvement of SULT1A3 in elevated sulfation of 4-hydroxypropranolol in Hep G2 cells pretreated with beta-naphthoflavone. AB - Pretreatment of Hep G2 cells with beta-naphthoflavone (BNF 1-25microM) significantly increased cytosolic sulfation activities of 4-hydroxypropranolol (4 OH-PL) racemate. The profile was similar to those of sulfations towards dopamine and triiodothyronine in the same cytosolic fractions. Kinetic studies of 4-OH-PL sulfation in Hep G2 cytosolic fractions revealed that V(max) values increased but apparent K(m) values remained unchanged following the BNF pretreatment. Among five recombinant human SULT isoforms (SULT1A1, -1A3, -1B1, -1E1 and -2A1) examined, only SULT2A1 did not show 4-OH-PL sulfation activities under the conditions used. SULT1A3 and -1E1 exhibited an enantioselectivity of 4-OH-R-PL sulfation>4-OH-S-PL sulfation, which agreed with that of BNF-pretreated Hep G2 cells as well as of nontreated cells, whereas SULT1A1 and -1B1 showed a reversed enantioselectivity (R S as the group VIII atom > distance separating the N and group VIII atom, as the major determinants contributing to the substrate behavior for low-molecular weight amine substrates of TG 2. PMID- 15748708 TI - L-arginine regulates neuronal nitric oxide synthase production of superoxide and hydrogen peroxide. AB - Tetrahydrobiopterin (H(4)B) in the absence of L-arginine has been shown to be an important factor in promoting the direct formation of hydrogen peroxide (H(2)O(2)) at the expense of superoxide (O(2)(*-)) by neuronal nitric oxide synthase (NOS1) [Rosen GM, Tsai P, Weaver J, Porasuphatana S, Roman LJ, Starkov AA, et al. Role of tetrahydrobiopterin in the regulation of neuronal nitric-oxide synthase-generated superoxide. J Biol Chem 2002;277:40275-80]. Based on these findings, it is hypothesized that L-arginine also shifts the equilibrium between O(2)(*-) and H(2)O(2). Experiments were designed to test this theory. As the concentration of L-arginine and N(omega)-hydroxyl-L-arginine increases, the rate of NADPH consumption for H(4)B-bound NOS1 decreased resulting in lower rates of both O(2)(*-) and H(2)O(2) generation, while increasing the rate of nitric oxide (*NO) production. At saturating concentrations of L-arginine or N(omega)-hydroxyl L-arginine (50microM), NOS1 still produced O(2)(*-) and H(2)O(2). Both L-arginine and N(omega)-hydroxyl-L-arginine have greater impact on the rate of generation of O(2)(*-) than on H(2)O(2). PMID- 15748709 TI - Biodistribution and metabolism of immunostimulatory oligodeoxynucleotide CPG 7909 in mouse and rat tissues following subcutaneous administration. AB - To evaluate pharmacokinetics (PK) and biodistribution, CPG 7909, a 24-mer immunostimulatory fully phosphorothioated oligodeoxynucleotide (PS-ODN), was administered by subcutaneous injection at 2, 5 and 12.5mg/kg to mice and at 9mg/kg to rats. Parent compound and metabolites were isolated from plasma and tissues and quantified by capillary gel electrophoresis with UV detection (CGE UV) and molecular masses were determined by matrix-assisted-laser-desorption ionization time of flight detection (MALDI-TOF). An established method for PS-ODN isolation from plasma and tissue was modified to prevent oxidation of the phosphorothioate bonds during the extraction process, significantly increasing sensitivity in the subsequent MALDI-TOF analysis. Concentrations of CPG 7909 and metabolites were highest at the injection site (>600mg/kg at 4h). Maximal concentrations in local (draining) lymph nodes (LLN), kidney and liver were 10 15% of that at the injection site. The highest total amount of PS-ODN (percentage of administered dose) was found in the liver (32% at 4h), followed closely by the injection site (23% at 4h). Only very low levels of CPG 7909 and metabolites were found in plasma and only during the first hours. Metabolites identified by MALDI TOF were similar for both species and all analyzed tissues, although the relative amounts of the different metabolites varied with tissue and over time. Degradation of CPG 7909 in vivo occurred predominantly by 3'exonucleases with additional cleavage by endonucleases. PMID- 15748710 TI - Restored expression and activity of organic ion transporters rOAT1, rOAT3 and rOCT2 after hyperuricemia in the rat kidney. AB - We previously reported that in hyperuricemic rats, renal impairment occurred and organic ion transport activity decreased, accompanied with a specific decrease in the expression of rat organic anion transporters, rOAT1 and rOAT3, and organic cation transporter, rOCT2. In the present study, we investigated the reversibility of the organic ion transport activity and expression of organic ion transporters (slc22a) during recovery from hyperuricemia. Hyperuricemia was induced by the administration of a chow containing uric acid and oxonic acid, an inhibitor of uric acid metabolism. Four days after discontinuance of the chow, the plasma uric acid concentration returned to the normal level, and renal functions such as creatinine clearance and BUN levels were restored, although the recovery of tubulointerstitial injury was varied in sites of the kidney. Basolateral uptake of p-aminohippurate (PAH) and tetraethylammonium (TEA), and both protein and mRNA levels of rOAT1, rOAT3 and rOCT2 in the kidney gradually improved during 14 days of recovery from hyperuricemia. Basolateral PAH transport showed a higher correlation with the protein level of rOAT1 (r(2)=0.80) than rOAT3 (r(2)=0.34), whereas basolateral TEA transport showed a strong correlation with rOCT2 protein (r(2)=0.91). The plasma testosterone concentration, which is a dominant factor in the regulation of rOCT2, was gradually restored during the recovery from hyperuricemia, but the correlation between the plasma testosterone level and rOCT2 protein expression in the kidney was not significant. These results suggest that the regulation of organic ion transporters, rOAT1, rOAT3 and rOCT2, by hyperuricemia is reversible, and the organic ion transport activity restores according to the expression levels of these transporters. PMID- 15748711 TI - Two types of aromatase with different encoding genes, tissue distribution and developmental expression in Nile tilapia (Oreochromis niloticus). AB - We isolated a novel type of aromatase cDNA from a Nile tilapia (Oreochromis niloticus) ovary cDNA library. Because this aromatase is phylogenetically related to brain aromatase (CYP19b) of goldfish, zebrafish and sea bass, we named it tilapia CYP19b (tCYP19b). tCYP19b encodes a protein that is predicted to consist of 495 residues and have 63.8% homology with the aromatase (tCYP19a) we previously isolated from the same source. In vitro transient transfection of cultured COS7 cells demonstrated that tCYP19b codes a functional protein to catalyze estrogen production from an androgen substrate. RT-PCR and Northern hybridization analysis showed that tCYP19b was expressed at a high level in the brain and at a low level in a wide variety of other tissues, whereas tCYP19a was mainly present in the ovary and its level significantly increased during the vitellogenic stage. RT-PCR also detected tCYP19b expression in brain and gonad tissues of both female and male tilapia during sex differentiation, but tCYP19a was only found in the ovary of the fry at that period. These results suggest that tCYP19a plays a key role in sex differentiation and ovarian development. We also isolated genes of two tilapia aromatases. Based on the location of the transcription initiation site, we predicted that there is one promoter for tCYP19a and three promoters for tCYP19b. Although the two aromatase isoforms have similar gene structures in the coding region, we found that the binding regions of SF-1/Ad4 BP region, WT1-KTS and SRY, which are sex-determining factors in mammals, are present in the 5' flank region of tCYP19a but not tCYP19b. A similar situation is present in promoters of zebrafish and goldfish aromatase isoforms. This data indicates that CYP19a plays a decisive role in sex differentiation of those species. The unique presence of the ERE motif in the tCYP19b promoter and the high expression of tCYP19b in the brain support that CYP19b is mainly involved in estrogen-mediated neural estrogen synthesis. PMID- 15748712 TI - Estrogen regulation of in vitro brain tyrosine hydroxylase activity in the catfish Heteropneustes fossilis: interactions with cAMP-protein kinase A and protein kinase C systems in enzyme activation. AB - In the present in vitro study, interactions of both cAMP-protein kinase A (PKA) and protein kinase C (PKC) systems were investigated in the estradiol-17beta (E2) regulation of forebrain (hypothalamus and telencephalon) tyrosine hydroxylase (TH) activity in the female catfish Heteropneustes fossilis in vitellogenic phase. E2 produced biphasic effects on TH activity: low concentrations (10(-12) 10(-5) M) stimulated, and high concentrations (10(-3)-10(-4) M) inhibited enzyme activity (Tukey's test, P<0.05). Co-incubations of the enzyme preparations with cAMP (1.0 mM), IBMX (1.5 mM) or theophylline (1.5 mM) and a low concentration of E2 (10(-9) M) increased TH activity significantly. However, the co-incubations with a high concentration of E2 (10(-3) M) decreased it significantly. Pre incubations of the enzyme preparations with cAMP (0.1 mM), followed by different concentrations of E2 (10(-12), 10(-9), 10(-4), and 10(-3) M) produced concentration-dependent biphasic effects. The pre-incubations with a low concentration of E2 (10(-9) M), followed by different concentrations of cAMP (0.05-1.0 mM) produced a significant concentration-dependent stimulation of TH activity and that with a high concentration of E2 (10(-3) M) produced a significant decrease in TH activity. Co-incubations of high and low E2, with or without cAMP, and PKA inhibitor (H-89) decreased TH activity significantly. The incubations with H-89 abolished the stimulatory effect of low E2 or low E2+cAMP and intensified the inhibitory effect of high E2 or high E2+cAMP combination. Co incubations with PKC inhibitor (calphostin C) did not influence the stimulatory effect of low E2 but lowered the stimulatory effect of low E2+cAMP treatment. Kinetic studies showed that the stimulatory effect of a low E2 concentration was due to a decrease in apparent Km and an increase in apparent Vmax for both cofactor and substrate, and the inhibitory effect of a high E2 concentration was due to reverse changes in the kinetics. The stimulatory effect of cAMP alone or in combination with low E2 was related to decreased Km and increased Vmax for the cofactor. The inhibitory effect of PKA and PKC blockers, alone or in combination with E2 and/or cAMP was due to increased Km and decreased Vmax of the enzyme for the cofactor. The present data suggest that E2 modulates the short-term activation of brain TH activity differentially and may involve mainly the cAMP PKA system. PMID- 15748713 TI - Effects of external pH on hormonally regulated ovarian follicle maturation and ovulation in Atlantic croaker. AB - In vitro studies of ovarian follicle maturation and ovulation in teleost fishes typically are conducted within a narrow range (7.5-7.8) of constant external (medium) pH, although there is evidence that pH can influence ovulation. Therefore, this study with Atlantic croaker investigated the effects of external pH on hormonally regulated in vitro maturation and ovulation as well as changes in the pH of ovarian fluid during in vivo maturation and ovulation. For the in vitro experiments, follicles were first incubated with human chorionic gonadotropin (hCG) to induce maturational and ovulatory competencies, and then with maturation-inducing hormone (MIH) to induce completion of maturation and ovulation. At a constant external pH within the range of 7.0-8.2, the lower pH levels (7.0-7.3) generally inhibited or slowed down hormonally induced maturation and ovulation whereas higher pH (7.6-8.2) facilitated these processes. When ovarian follicles were incubated at a constant pH of 7.6 during the priming incubation with hCG, changing the external pH during the incubation with MIH had relatively little effect on oocyte maturation or ovulation. Thus, the inhibitory effect of constant low levels of external pH (7.0-7.3) on maturation and ovulation may be primarily due to disruptions in the gonadotropin-dependent acquisition of maturational and ovulatory competencies. The pH of ovarian fluid remained constant at 8.5 during in vivo ovarian follicle maturation and ovulation. Subsequent in vitro tests showed that external pH of 8.5 enhances hormonally induced maturation and ovulation relative to pH of 7.6. These observations suggest that attention should be paid to the pH of incubation media used in basic research and in biotechnological applications relying on in vitro maturation and ovulation in teleosts. Further, an understanding of the physiological significance of the enhancing effect of alkaline pH on maturation and ovulation will require determination of the intrafollicular pH around the oocyte during the periovulatory period. PMID- 15748714 TI - Influence of hormonal status on enkephalin-degrading aminopeptidase activity in the HPA axis of female mice. AB - Opioids are involved in the regulation of hypothalamus-pituitary-adrenal (HPA) axis activity under physiological conditions. In the present work, we analyzed the influence of ovariectomy and estradiol (E), progesterone (P) or estradiol plus progesterone (E+P) replacement on soluble (S) and membrane-bound (MB) enkephalin-degrading aminopeptidase activity (EDA) in the HPA axis. Female mice (Balb/C) were distributed in 15 groups of 10 animals each: sham-operated controls (C), ovariectomized controls (OV-C), and ovariectomized mice treated with increasing doses of E (10, 20 or 40 mg/kg), P (100, 200 or 400 mg/kg) or E+P (10+100, 20+200 or 40+400 mg/kg). In hypothalamus, ovariectomy increased both S and MB EDA activities, whereas E replacement returned them to control levels, although MB EDA activity increased again after the replacement with 40 mg/kg E. P replacement increased S EDA activity, but returned MB EDA activity to control levels. The replacement of E+P returned both S and MB EDA activities to control levels, although MB EDA activity was lower than control values after the replacement with 10+100 mg/kg E+P. In pituitary, neither ovariectomy nor the replacement of E or E+P changed S EDA, although the highest concentrations of P increased S EDA activity. However, ovariectomy increased MB EDA and E replacement returned the activity to control or below control levels, depending on the concentration used. However, P administration returned the activity to control or below control levels depending on the concentration used, although 200 mg/kg P had no effects on MB EDA. E+P replacement returned pituitary MB EDA activity to control levels. In adrenal glands, ovariectomy did change either S or MB EDA. However, E, P or E+P replacement decreased S EDA activity in different degrees, depending of the dose administrated. No changes were detected in MB EDA after hormone replacement. These results indicate that female steroid hormones influence EDA activity at different levels of HPA axis. PMID- 15748715 TI - Studies on retinol-binding protein during vitellogenesis in the Rainbow Trout (Oncorhynchus mykiss). AB - Retinoids are important regulatory signaling molecules during embryonic development and therefore, should be present in the eggs of oviparous animals that develop independently of the maternal organism. Studies were initiated in Rainbow Trout to elucidate the role of retinol-binding protein (RBP), the specific retinol carrier protein in vertebrate plasma, during vitellogenesis. Plasma levels of RBP in pre-vitellogenic, vitellogenic, and post-vitellogenic females were compared to plasma of male trout, using a rabbit polyclonal antiserum that was generated to His-tagged RBP recombinant protein. Western-blot analyses showed that there were no differences in the relative plasma levels of RBP between pre-vitellogenic, vitellogenic or post-vitellogenic females and similar levels were also found in males. In contrast, strong elevation of vitellogenin (VTG) was observed in the plasma from vitellogenic females. Northern blot analysis of hepatic mRNA revealed that there were no dramatic changes in the abundance of RBP transcripts in the liver of females during vitellogenesis, but showed a significant increase in the expression of VTG in the livers of vitellogenic females. These results indicate differences in the regulation of RBP and VTG during vitellogenesis, suggesting that RBP may not be the main transporter protein for retinoids to fish egg. Recent publications on the association of retinal with VTG in fish and the occurrence of RBP transcripts in ovarian tissues raise the need for reevaluation of the role of RBP during vitellogenesis in oviparous non-mammalian vertebrate species. PMID- 15748717 TI - Ontogeny of GnRH-like immunoreactive neuronal systems in the forebrain of the Indian major carp, Cirrhinus mrigala. AB - GnRH immunoreactivity appeared in the medial olfactory placode very early in the development of Cirrhinus mrigala. The immunoreactive elements were divisible into distinct migratory and non-migratory components. The migratory component appeared as a patch of intensely immunoreactive cells located close to the olfactory epithelium in day 6 post-fertilization larvae. Subsequently, these neurons migrate caudally along the ventromedial aspect of the developing forebrain and enroute give rise to GnRH immunoreactive neurons in the (1) nervus terminalis located in ventral and caudal part of the olfactory bulb (day 8), and (2) basal telencephalon (day 9). The non-migratory GnRH immunoreactive component appeared in the olfactory placode of day 1 post-fertilization larvae. It consisted of few olfactory receptor neuron (ORN)-like cells with distinct flask-shaped somata, dendrites that communicate with the periphery and a single axon on the basal side; GnRH immunoreactivity was seen throughout the neuron. Considerable increase in the number of immunoreactive ORNs was encountered in day 2 post-fertilization larvae. On day 3, the dendrites of ORNs sprout bunches of apical cilia, while on the basal side the axonal outgrowths can be traced to the olfactory bulb. GnRH immunoreactive fibers were distributed in the olfactory nerve layer in the periphery of the bulb and glomeruli-like innervation was clearly established in 5 days old larvae. The innervation to the olfactory bulb showed a considerable increase in GnRH immunoreactivity in 9 and 19 days old larvae. However, GnRH immunoreactivity in non-migratory as well as migratory components gradually diminished and disappeared altogether by the age of 68 days. Results of the present study suggest that GnRH may serve a neurotransmitter role in the ORNs during early stages of development in C. mrigala. PMID- 15748716 TI - Steroid-induced changes in the morphology of GnRH neurons in the male leopard frog, Rana pipiens: correlation with plasma gonadotropin and gonadal size. AB - Previously, we reported that hypothalamic explants isolated from male leopard frogs (Rana pipiens) implanted with 17beta-estradiol (E2), but not 5alpha dihydrotestosterone (DHT), released significantly higher levels of gonadotropin releasing hormone (GnRH) in response to a veratridine challenge. In this study, we measured changes in GnRH soma size, circulating luteinizing hormone (LH), and gonadosomatic index (GSI) in response to these two steroid hormones to further assess the impact of these hormones on the hypothalamic-pituitary-gonadal axis. Sexually mature male R. pipiens were implanted with silastic capsules containing cholesterol (Ch; control), E2, or DHT for 20 days. GnRH immunocytochemistry (ICC) revealed that both E2 and DHT significantly enlarged GnRH soma size without affecting the total number of GnRH neurons in the forebrain. The effects of E2 and DHT were specific, since neither hormone altered the soma size of tyrosine hydroxylase neurons in the dorsomedial posterior tuberculum. Circulating LH levels were significantly reduced in animals treated with both steroid hormones, with E2 exerting the most potent inhibitory effect. A significant inverse correlation was observed between the GSI and GnRH soma size in Ch controls, suggesting animals with larger GnRH neurons tended to have smaller gonads. Overall, our results showed that both steroid hormones induced the accumulation of GnRH and ultimately the swelling of the GnRH soma. Further, larger GnRH neurons were associated with smaller gonads and lower circulating levels of LH, suggesting a link between enlarged GnRH neurons and an overall decrease in the reproductive activity of R. pipiens. PMID- 15748718 TI - Prolactin acts centrally to enhance newt courtship behavior. AB - The effects of intracerebroventricular (ICV) and intraperitoneal (IP) injections of ovine prolactin (PRL), antiserum against newt PRL, and antibody against the newt PRL receptor on the expression of courtship behavior of male newts, Cynops pyrrhogaster, were studied to see whether PRL acts centrally or peripherally to induce the behavior. Injections of PRL by either route into gonadotropin-primed males enhanced the expression of the behavior dose-dependently. The minimum effective amount of ovine PRL administered intracerebroventricularly was 0.1 microg, whereas it was 100 microg when injected intraperitoneally. ICV injection of antiserum against newt PRL blocked the spontaneously occurring male courtship behavior when the anti-newt PRL serum was given either intracerebroventricularly or intraperitoneally. The minimum effective dose of the antiserum administered intracerebroventricularly was 0.05 microl, whereas it was 20 microl when injected intraperitoneally. Neither ICV nor IP injection of preimmune serum affected the expression of the behavior. Furthermore, ICV, but not IP, administration of 0.3 microg of anti-newt PRL receptor antibody, purified from antiserum against newt PRL receptor by use of an antigen-conjugated affinity column, blocked the spontaneously occurring courtship behavior in sexually developed males. Neither ICV nor IP injection of the same amount of normal rabbit IgG affected the expression of the behavior. The results strongly suggest that endogenous PRL enhances the behavior by acting centrally through the PRL receptors localized in the brain area. PMID- 15748720 TI - Production of symmetrical flatfish by controlling the timing of thyroid hormone treatment in spotted halibut Verasper variegatus. AB - The thyroid hormone plays an essential role in the metamorphosis in flatfish, during which external asymmetry (for example, eye relocation and pigmentation) is established. However, no information is available on the expression mechanisms of metamorphic asymmetry. Since the presence of malformed juveniles (having either ocular or blind side characteristics on both sides) was known in spotted halibut (Verasper variegatus), the effect of the timing of thyroid hormone treatment was investigated. When thyroxine (T4, 10 or 30 ng/ml) was administered to the hypothyroid larvae (continuously receiving 30 microg/ml thiourea) from 25 days after hatching (DAH), the occurrence of one type of symmetrical juvenile (symmetrical pseudoalbino having blind side characteristics on both sides) increased to more than three times, and the occurrence was significantly greater than that observed in the control group. In the fish in which T4 treatment was initiated prior to 15 DAH or after 60 DAH, the occurrence of another type of symmetrical juvenile (symmetrical ambicolorate having ocular side characteristics on both sides) became more than two times, and its occurrence was significantly greater than that observed in the control group. These results suggest that both sides of the larval body independently possess the potential to become either the ocular or the blind side of the juvenile body and that the timing of thyroid hormone increase is the determining factor of "which side to become." Based on previous studies, since thyroid hormone is expected to induce blind side characteristics, the differential responsiveness to thyroid hormone-longer in the left side (blind side in normal juvenile) than the right-is strongly suggested as the central mechanism for metamorphic asymmetry in spotted halibut. PMID- 15748719 TI - Different effects of peripheral and central ghrelin on regulation of food intake in the Japanese quail. AB - When rat or human ghrelin is administered to chickens or rats it stimulates the release of growth hormone, however, its effects on food intake differ between the two species. To investigate this discrepancy, we measured plasma ghrelin concentrations before and after food intake, and determined the effects of central (intracerebroventricular, icv) and peripheral (intraperitoneal, ip) injections of various ghrelin doses on food intake and body temperature in the Japanese quail. In control quails, plasma ghrelin levels were significantly increased in the fasting state; subsequent feeding produced a reduction, suggesting that ghrelin may act as an orexigenic signal in Japanese quails as well as mammals. Food intake was stimulated by ip, but not icv, injections of small doses of ghrelin, whereas both ip and icv injections of larger doses inhibited feeding. A large dose of ghrelin also increased body temperature. These results suggest that an increase in peripheral ghrelin may act as a hunger signal to induce food intake through stimulation of the afferent vagal nerve in the Japanese quail, as in rats, whereas central ghrelin may inhibit feeding in different ways in the two species. PMID- 15748721 TI - Molecular cloning of the cDNA encoding follicle-stimulating hormone beta subunit of the Chinese soft-shell turtle Pelodiscus sinensis, and its gene expression. AB - Follicle-stimulating hormone (FSH) is a member of the pituitary glycoprotein hormone family. These hormones are composed of two dissimilar subunits, alpha and beta. Very little information is available regarding the nucleotide and amino acid sequence of FSHbeta in reptilian species. For better understanding of the phylogenetic diversity and evolution of FSH molecule, we have isolated and sequenced the complementary DNA (cDNA) encoding the Chinese soft-shell turtle (Pelodiscus sinensis, Family of Trionychidae) FSHbeta precursor molecule by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) methods. The cloned Chinese soft-shell turtle FSHbeta cDNA consists of 602-bp nucleotides, including 34-bp nucleotides of the 5' untranslated region (UTR), 396-bp of the open reading frame, and 3'-UTR of 206-bp nucleotides. It encodes a 131-amino acid precursor molecule of FSHbeta subunit with a signal peptide of 20 amino acids followed by a mature protein of 111 amino acids. Twelve cysteine residues, forming six disulfide bonds within beta-subunit and two putative asparagine-linked glycosylation sites, are also conserved in the Chinese soft-shell turtle FSHbeta subunit. The deduced amino acid sequence of the Chinese soft-shell turtle FSHbeta shares identities of 97% with Reeves's turtle (Family of Bataguridae), 83-89% with birds, 61-70% with mammals, 63-66% with amphibians and 40-58% with fish. By contrast, when comparing the FSHbeta with the beta-subunits of the Chinese soft-shell turtle luteinizing hormone and thyroid stimulating hormone, the homologies are as low as 38 and 39%, respectively. A phylogenetic tree including reptilian species of FSHbeta subunits, is presented for the first time. Out of various tissues examined, FSHbeta mRNA was only expressed in the pituitary gland and can be up-regulated by gonadotropin releasing hormone in pituitary tissue culture as estimated by fluorescence real time PCR analysis. PMID- 15748722 TI - Perceptual ratings of vocal characteristics and voicing features in untreated patients with unilateral vocal fold paralysis. AB - This study used visual analog scales to obtain perceptual ratings of features of voice production in subjects with unilateral vocal fold paralysis (UVFP), including clarity of laryngeal articulation, consistency of loudness across the utterance and the voiced/voiceless distinction. Recordings of repeated /i/, /isi/, and /izi/ from subjects diagnosed with UVFP and control subjects were randomly re-recorded, and then rated by five listeners. Significant differences in ratings (Smirnov test, p < 0.01) were found between groups for "aphonia", "severity", "clarity of articulation", "overall loudness", "consistency of loudness" and "amount of effort". Four of five raters agreed on the accuracy of /s/ or /z/ productions for only 54% of the samples from the subjects with UVFP. Voiceless and voiced cognates were equally likely to be rated as inaccurate. Results suggested that these variables were sensitive to changes in voice production resulting from paralysis, and may be useful in measuring treatment outcomes and spontaneous recovery of function. LEARNING OUTCOMES: As a result of reading this manuscript the reader will (1) gain an understanding of types of perceptual scales and how to develop the set of vocal characteristics to be used in distinguishing patients with UVFP and those without, (2) learn which vocal characteristics listeners are able to use to successfully distinguish between patients with UVFP and those without and (3) understand the possible role for perceptual ratings in tracking changes in vocal characteristics in subjects over time following treatment or spontaneous recovery of function. PMID- 15748723 TI - The effects of levodopa on word intelligibility in Parkinson's disease. AB - Dysarthria is a common manifestation in patients with idiopathic Parkinson's disease. This study investigated the effects of levodopa on intelligibility in patients with Parkinson's disease. Ten participants were tested during on- and off-states using the Yorkston and Beukelman intelligibility test (1980). Intelligibility as scored by a panel of speech therapists was significantly improved in the on-condition. No correlation was found, however, between intelligibility and overall severity of the disease or severity of the motor problems. EDUCATIONAL OUTCOMES: As a result of this activity the participant will be able to discuss the effects of levodopa on intelligibility in patients with Parkinson's disease. PMID- 15748724 TI - Language sampling for kindergarten children with and without SLI: mean length of utterance, IPSYN, and NDW. AB - Language sample analysis measures have long been promoted as exhibiting greater ecological validity than formal testing in the assessment of language disorder in children. In practice, their use is often restricted to preschool children, owing to lack of normative information, as well as criticisms of the validity of commonly used measures for the language of older children. This study compared scores of kindergarten children (mean age 6 years) with and without specific language impairment (SLI) on three commonly used language sample analysis measures: mean length of utterance in morphemes (MLU-m), the index of productive syntax (IPSyn), and number of different words (NDWs). Mean scores of the children with SLI were significantly lower for all three measures, though not for all subtests of the IPSyn. A number of individual differences were observed; notably, several children with SLI scored as well as those without. The problems and promise of language sampling for children beyond the preschool years are discussed in light of these results. LEARNING OUTCOMES: (1) Readers will gain an understanding of strengths and weaknesses of language sample measures in assessing kindergarten children with language impairment. (2) The reader will become aware of the utility of MLU in differentiating between young school age children with and without language impairment. PMID- 15748725 TI - Acoustic analysis of clear versus conversational speech in individuals with Parkinson disease. AB - A number of studies have been devoted to the examination of clear versus conversational speech in non-impaired speakers. The purpose of these previous studies has been primarily to help increase speech intelligibility for the benefit of hearing-impaired listeners. The goal of the present study was to examine differences between conversational and clear speech in individuals with Parkinson disease (PD). Twelve individuals were recorded producing conversational and clear speech. Acoustic analysis revealed that individuals with PD used some of the same clear speech strategies used by non-impaired speakers. Specifically, clear speech in PD was characterized by decreased articulation rate, increased mean fundamental frequency (F(o)), and increased speaking F(o)S.D. compared to conversational speech. The discussion examines the possibility that individuals with PD may have been independently applying a clear speech strategy based on their habitually increased percent pause values. Discussion also focuses on implications of the present findings to management of individuals with PD, and research implications. LEARNING OUTCOMES: As a result of this activity, the participant will be able to (1) describe the characteristics of clear speech produced by non-disordered individuals, (2) describe the acoustic characteristics of clear and conversational speech produced by individuals with Parkinson disease; and (3) describe the strategies individuals with Parkinson disease use when asked to produce clear speech. PMID- 15748726 TI - Dysarthria associated with traumatic brain injury: speaking rate and emphatic stress. AB - Prosodic abnormality is common in the dysarthria associated with traumatic brain injury (TBI), and adjustments of speaking rate and emphatic stress are often used as steps in treating the speech disorder in patients with TBI-induced dysarthria. However, studies to date do not present a clear and detailed picture of how speaking rate and emphatic stress are affected in this speech disorder. This study, based on the acoustic analyses of syllable repetitions and sentence speech samples, reports on speaking rate and emphatic stress for 12 subjects with TBI and 8 healthy controls. For speaking rate, the subjects with TBI had (1) both slow speaking and articulation rates, (2) smaller phonation proportion and larger pause proportion, and (3) larger percentage change in speaking rate and smaller percentage change in articulation rate. For emphatic stress, the subjects with TBI had (1) significant increases in the difference and percentage change between pre-stressed and pre-unstressed pause durations, (2) significantly smaller difference between stressed and unstressed word durations, but not the percentage change between stressed and unstressed word durations, and (3) significantly reduced differences in f(0) movement and f(0) slope between stressed and unstressed words, but not in RMS range. This study demonstrates the multidimensional nature of prosodic deficits in the dysarthria related to TBI and illustrates the ability of acoustic measures to give a picture of the dysprosody related to TBI-induced dysarthria. LEARNING OUTCOMES: As a result of this activity, the participant will be able to (1) describe the prosodic disturbances that have been reported in studies of dysarthria associated with TBI; (2) define acoustic measures appropriate to the analysis of changes in speaking rate and emphatic stress; and (3) discuss the importance of prosody to spoken communication. PMID- 15748727 TI - Hidden Markov analysis of mechanosensitive ion channel gating. AB - Patch clamp data from the large conductance mechanosensitive channel (MscL) in E. coli was studied with the aim of developing a strategy for statistical analysis based on hidden Markov models (HMMs) and determining the number of conductance levels of the channel, together with mean current, mean dwell time and equilibrium probability of occupancy for each level. The models incorporated state-dependent white noise and moving average adjustment for filtering, with maximum likelihood parameter estimates obtained using an EM (expectation maximisation) based iteration. Adjustment for filtering was included as it could be expected that the electronic filter used in recording would have a major effect on obviously brief intermediate conductance level sojourns. Preliminary data analysis revealed that the brevity of intermediate level sojourns caused difficulties in assignment of data points to levels as a result of over estimation of noise variances. When reasonable constraints were placed on these variances using the better determined noise variances for the closed and fully open levels, idealisation anomalies were eliminated. Nevertheless, simulations suggested that mean sojourn times for the intermediate levels were still considerably over-estimated, and that recording bandwidth was a major limitation; improved results were obtained with higher bandwidth data (10 kHz sampled at 25 kHz). The simplest model consistent with these data had four open conductance levels, intermediate levels being approximately 20%, 51% and 74% of fully open. The mean lifetime at the fully open level was about 1 ms; estimates for the three intermediate levels were 54-92 micros, probably still over-estimates. PMID- 15748728 TI - When do mixotrophs specialize? Adaptive dynamics theory applied to a dynamic energy budget model. AB - In evolutionary history, several events have occurred at which mixotrophs specialized into pure autotrophs and heterotrophs. We studied the conditions under which such events take place, using the Dynamic Energy Budget (DEB) theory for physiological rules of the organisms' metabolism and Adaptive Dynamics (AD) theory for evolutionary behavior of parameter values. We modeled a population of mixotrophs that can take up dissolved inorganic nutrients by autotrophic assimilation and detritus by heterotrophic assimilation. The organisms have a certain affinity for both pathways; mutations that occur in the affinities enable the population to evolve. One of the possible evolutionary outcomes is a branching point which provides an opportunity for the mixotrophic population to split up and specialize into separate autotrophs and heterotrophs. Evolutionary branching is not a common feature of the studied system, but is found to occur only under specific conditions. These conditions depend on intrinsic properties such as the cost function, the level of the costs and the boundaries of the trait space: only at intermediate cost levels and when an explicit advantage exists to pure strategies over mixed ones may evolutionary branching occur. Usually, such an advantage (and hence evolutionary branching) can be induced by interference between the two affinities, but this result changes due to the constraints on the affinities. Now, only some of the more complicated cost functions give rise to a branching point. In contrast to the intrinsic properties, extrinsic properties such as the total nutrient content or light intensity were found to have no effect on the evolutionary outcomes at all. PMID- 15748729 TI - Biofilms and the plasmid maintenance question. AB - Can a conjugative plasmid encoding enhanced biofilm forming abilities for its bacterial host facilitate the persistence of the plasmid in a bacterial population despite conferring diminished growth rate and segregative plasmid loss on its bearers? We construct a mathematical model in a chemostat and in a plug flow environment to answer this question. Explicit conditions for an affirmative answer are derived. Numerical simulations support the conclusion. PMID- 15748730 TI - Controlling emerging infectious diseases like SARS. AB - To control emerging infectious diseases like SARS, it is necessary to resort to basic control measures that limit exposures to infectious individuals. These measures include isolating cases at diagnosis, quarantining household members and tracing contacts of diagnosed cases, providing the community with advice on how to reduce exposures, and closing schools. To justify such intervention it is important to understand how well each of these measures helps to limit transmission. In this paper, we determine the effect of a number of different interventions on the effective reproduction number and estimate requirements to achieve elimination of the infectious disease. We find that the strategy of tracing and quarantining contacts of diagnosed cases can be very successful in reducing transmission. PMID- 15748731 TI - Percolation of annotation errors through hierarchically structured protein sequence databases. AB - Databases of protein sequences have grown rapidly in recent years as a result of genome sequencing projects. Annotating protein sequences with descriptions of their biological function ideally requires careful experimentation, but this work lags far behind. Instead, biological function is often imputed by copying annotations from similar protein sequences. This gives rise to annotation errors, and more seriously, to chains of misannotation. [Percolation of annotation errors in a database of protein sequences (2002)] developed a probabilistic framework for exploring the consequences of this percolation of errors through protein databases, and applied their theory to a simple database model. Here we apply the theory to hierarchically structured protein sequence databases, and draw conclusions about database quality at different levels of the hierarchy. PMID- 15748732 TI - Interval observers for biochemical processes with uncertain kinetics and inputs. AB - This paper concentrates on the state observation in bioprocesses when there is uncertainty on the process parameters and/or the process inputs. An interval observer is designed on the basis of the cooperativity properties of the model for a standard stirred tank bioreactor model with a single microbial growth and a kinetic model depending on the substrate concentration. Further assumptions are the (lower and upper) boundedness of the specific growth rate and the inlet substrate concentration. Mathematical analysis of the stability and convergence of the interval observer is performed both in absence and in presence of uncertainty on the measurements. It is shown in particular that when the process inputs are known, the static observation error on the unknown state is inversely proportional to one of the observer gains. The performance of the interval observer are also illustrated through numerical simulation. PMID- 15748733 TI - Estimating the life-span of oligodendrocytes from clonal data on their development in cell culture. AB - This paper presents a new method to analyze clonal data on oligodendrocyte development in cell culture. The process of oligodendrocyte generation from precursor cells is modelled as a multi-type Bellman-Harris branching process as suggested in an earlier paper [K. Boucher, A. Zorin, A.Y. Yakovlev, M. Mayer Proschel, M. Noble, An alternative stochastic model of generation of oligodendrocytes in cell culture, J. Math. Biol. 43 (2001) 22]. This model has been extended to allow for death of oligodendrocytes as well as a dissimilar distribution of the first mitotic cycle duration as compared to the subsequent cycles of precursor cells, which lengths are assumed to be independent and identically distributed random variables. Since the time-span of oligodendrocytes is not directly observable in clonal data, plausible parametric assumptions are invoked to make estimation problems tractable. In particular, the time to cell death follows a two-parameter gamma distribution, while the lapse of time between the event of cell death and the event of cell disintegration is assumed to be exponentially distributed. A simulated pseudo maximum likelihood method for estimation of model parameters has been developed using simulation-based approximations of the expected numbers and variance-covariance matrices for different types of cells. Finite sample properties of the estimation procedure are studied by computer simulations. The proposed method is illustrated with an analysis of the clonal development of O-2A progenitor cells isolated from the rat optic nerve and the corpus callosum. PMID- 15748734 TI - Immunocytochemical localization of tubulins in spermatids and spermatozoa of Euptoieta hegesia (Lepidoptera: Nymphalidae). AB - A comparative analysis of the distribution of tubulin types in apyrene and eupyrene sperm of Euptoieta hegesia butterflies was carried out, also verifying the presence of tubulin in lacinate appendages of the eupyrene sperm. Ultrathin sections of LR White embedded spermatids and spermatozoa were labeled for alpha, beta, gamma, alpha-acetylated and alpha-tyrosinated tubulins. Apyrene and eupyrene spermatids show the same antibody recognition pattern for tubulins. All tubulin types were detected in axonemal microtubules. Alpha and gamma tubulins were also detected on the cytoplasmic microtubules. However, for beta and tyrosinated tubulins only scattered labeling was detected on cytoplasmic microtubules and acetylated tubulin was not detected. In apyrene and eupyrene spermatozoa only the axoneme labeling was analyzed since cytoplasmic microtubules no longer exist in these cells. Alpha, beta and tyrosinated tubulins were easily detected on the apyrene and eupyrene axoneme; gamma tubulin was strongly marked on eupyrene axonemes but was scattered on the apyrene ones. Acetylated tubulin appeared with scattered labeling on the axoneme of both sperm types. Our results demonstrate significant differences in tubulin distribution in apyrene and eupyrene axonemal and cytoplasmic microtubules. Extracellular structures, especially the lacinate appendages, were not labeled by antibodies for any tubulin. PMID- 15748735 TI - Follicular diameter range based on morphological features in Synbranchus marmoratus (Bloch, 1795) (Teleostei, Synbranchiformes, Synbranchidae) from the South-central region of Brazil. AB - The morphological characteristics of Synbranchus marmoratus female germ cells in various development stages were described in details; then measurements of ovarian follicle diameters were taken from primary and secondary growth as during these development stages the oocyte size varied considerably along the fish growth. The results were correlated to total fish length, using the individuals division in six size classes. It was possible to group oocytes by stages according to histological characteristics but not according to morphometric diameter, as there was a wide variation in diameter in each stage and overlap between different maturation stages. These data make available new information on the reproductive biology of Synbranchidae. PMID- 15748736 TI - Reexamination of phenoloxidase in larval circulating hemocytes of the silkworm, Bombyx mori. AB - We have developed a modified method to detect phenoloxidase activity on hemocytes by using freshly prepared l-DOPA (1 mg/ml in 35% ethanol) to fix and incubate larval hemocytes. This method is more sensitive than the common method, in which hemocytes were fixed in 4% formaldehyde and then incubated with 2 mg/ml l-DOPA in water separately. Phenoloxidase assayed using this modified method can be inhibited by phenyltiourea (phenoloxidase inhibitor). After incubation with l DOPA solution in ethanol, most prohemocytes, all plasmatocytes and young granulocytes are stained brown due to oxidation of l-DOPA into pigments, indicating that they have phenoloxidase. Oenocytoids are dimly stained because many of their cell inclusions have been released during the treatment. Large propidium-iodide-negative prohemocytes have strong phenoloxidase activity and are easily misunderstood as propidium-iodide-positive oenocytoids if the fluorescent method is not used for identification. Thus, in addition to oenocytoids and plasmatocytes, some prohemocytes and granulocytes in the silkworm also have phenoloxidase. PMID- 15748737 TI - Spermatozoal ultrastructure of four Sparidae fishes: Acanthopagrus berda, Acanthopagrus australis, Lagodon rhomboids and Archosargus probatocephus. AB - The mature spermatozoa of two Taiwan protandrous hermaphrodite Sparidae Acanthopagrus berda and Acanthopagrus australis are investigated and compared with those of other two Sparidae (Lagodon rhomboids and Archosargus probatocephus) from the Western hemisphere. Ultrastructurally the spermatozoon of these four species has a spherical, homogeneously electron-dense nucleus with an axial nuclear fossa. The midpiece contains one to four spherical mitochondria and encircles the basal body of the flagellum. The mature spermatozoa of the four species are of the primitive or ect-aquasperm form and conform to the teleostean type I spermatozoon with the flagellar axis inserts perpendicular and medial to the nuclear fossa. Variation in the depths of the nuclear fossa and mitochondria number is substantial in these four Sparidae species. This study provide useful systematic characters to the existing knowledge of comparative spermatology of Sparidae. PMID- 15748738 TI - Ultrastructural studies of mosquito ovogenesis. AB - The ovogenesis of four mosquito species belonging to the genera Aedes, Anopheles and Culex, are investigated using Electron microscopes. Three ovogenetic phases named previtellogenesis, vitellogenesis, postvitellogenesis and mature eggs are described using transmission electron and light microscopes. Egg ornamentations are described with scanning electron microscopy. The controversial nomenclature of the mosquito egg envelopes is discussed. PMID- 15748739 TI - Morphology and ultrastructure of the earthworm Dendrobaena veneta (Lumbricidae) coelomocytes. AB - Microscope techniques, light microscope (LM), transmission electron microscope (TEM), scanning electron microscope (SEM) were employed to describe and classify coelomocytes of the oligochaete Dendrobaena veneta. Three main cell types were distinguished in the coelomic fluid: eleocytes, amoebocytes and granulocytes. Eleocytes are large, oval cells containing characteristic granules called chloragosomes. Amoebocytes are most numerous coelomocytes and have been divided into two types (I and II). Both amoebocytes of the types I and II often form aggregations of a few to about a dozen cells. Granulocytes are oval cells with spherical nuclei and cytoplasm containing polymorphic, electron dense granules. Contrary to the amoebocytes, the granulocytes do not form aggregations. Morphology and ultrastructure of coelomocytes are presented on micrographs: similarities and differences are compared to coelomocytes of related species. PMID- 15748740 TI - Midgut epithelium formation in Thermobia domestica (Packard) (Insecta, Zygentoma). AB - The origin of midgut epithelium may begin either from yolk cells (energids), tips of stomo- and proctodaeum (ectoderm), inner layer (endoderm) or from both kinds of the above mentioned cells. The origin of the midgut epithelium in wingless insects (Apterygota) has still not been determined. In Thermobia domestica the formation of midgut is much delayed, and it completes in the post-embryonic stage, while the stomo- and the proctodaeum are well-developed in the embryonic period. The energids, which remain inside the yolk, start to migrate to its periphery, where they arrange singly close to cell membrane. The yolk mass with the energids at the 14th day of embryogenesis are referred to as the primary midgut. During the first instar larval stage more and more energids migrate to the yolk periphery and the cell membrane starts to form numerous foldings surrounding the groups of energids, which in turn lead to formation of isolated regenerative cell groups. Eventually the cell membrane invaginations reach the center of the yolk mass. Large cells of the primary epithelium, surrounding the newly formed midgut lumen are formed. The cells of the primary epithelium are filled with yolk and are equipped with microvilli pointing to the midgut lumen. As the yolk is being digested, the process of the primary epithelium cells degeneration begins. The cells are getting shorter and start to degenerate. The definitive midgut epithelium is formed from proliferating regenerative cells. It consists of regularly spaced regenerative cell groups as well as the epithelial cells. The ultrastructure of both these cell groups has been described. PMID- 15748741 TI - Attachment of columnar airway epithelial cells in asthma. AB - Shedding of airway epithelial cells is a common finding in asthma. In this study, the attachment of the airway epithelial cells to the basal lamina (BL) was investigated by transmission electron microscopy (TEM) of biopsies from patients with atopic asthma and healthy controls. The following parameters were quantitatively determined: the height of the epithelium and of the columnar cells, the number of basal cells per 100 microm of basal lamina, the contact surfaces of basal cells or columnar cells with the basal lamina, and between basal cells and columnar cells. In order to compare the quantitative method with previous literature data, measurements were also carried out on rat airway epithelium. Compared to the rat, the columnar cell height in the human is increased, basal cells are smaller, and there is a larger contact area between basal cells and basal lamina, as well as between basal and columnar cells. The contact area between columnar cells and basal lamina is hence less in the human airway. The contact area between columnar cells and basal lamina in asthmatics is significantly less than in healthy controls, due to larger intercellular spaces. It is concluded that attachment of columnar cells to the basal lamina occurs mainly indirectly, via desmosomal attachment to basal cells, and that direct attachment of columnar cells to the basal lamina is weakened in asthmatics. PMID- 15748742 TI - Branchial chamber tissues in two caridean shrimps: the epibenthic Palaemon adspersus and the deep-sea hydrothermal Rimicaris exoculata. AB - The structure of the epithelia of the branchial chamber organs (gills, branchiostegites, epipodites) and the localization of the Na(+),K(+)-ATPase were investigated in two caridean shrimps, the epibenthic Palaemon adspersus and the deep-sea hydrothermal Rimicaris exoculata. The general organization of the phyllobranchiate gills, branchiostegites and epipodites is similar in P. adspersus and in R. exoculata. The gill filaments are formed by a single axial epithelium made of H-shaped cells with thin lateral expansions and a basal lamina limiting hemolymph lacunae. In P. adspersus, numerous ionocytes are present in the epipodites and in the inner-side of the branchiostegites; immunofluorescence reveals their high content in Na(+),K(+)-ATPase. In R. exoculata, typical ionocytes displaying a strong Na(+),K(+)-ATPase specific fluorescence are observed in the epipodites only. While the epipodites and the branchiostegites appear as the main site of osmoregulation in P. adspersus, only the epipodites might be involved in ion exchanges in R. exoculata. In both species, the gill filaments are mainly devoted to respiration. PMID- 15748743 TI - Introduction to the Special Issue on mechanisms in metal toxicology. PMID- 15748744 TI - Cadmium in the shore crab Carcinus maenas: seasonal variation in cadmium content and uptake and elimination of cadmium after administration via food. AB - The uptake and assimilation efficiency of cadmium administered via the food in the shore crab Carcinus maenas were investigated together with elimination kinetics and seasonal variations in cadmium content. The majority of shore crabs assimilated between 41 and 86% of the cadmium administered in their food. More than 90% of the cadmium taken up from food was retained in midgut gland. Elimination of cadmium after uptake from one meal of radioactively labelled soft parts of blue mussels could be described by a three-compartment model (percent 109Cd-retained = 64 x e(-0.001107 x t) + 25 x e(-0.0385 x t)+11 x e(-0.888 x t)). The biological half-life for cadmium in the most slowly exchanging compartment (containing 64% of the body burden) was 626 days. Groups of male and female shore crabs were collected from an uncontaminated site in the period May till October and the concentrations of cadmium in midgut gland and gills were determined. Male crabs had higher cadmium concentrations in the midgut gland in June and August (mean 2.7 microg Cd g(-1) dry weight) than they had in May, September and October (mean 1.7 microg Cd g(-1) dry weight). Females generally had slightly lower cadmium concentrations in the midgut gland than the males, except for a relatively high concentration in May. The cadmium concentrations in gills generally ranged between 0.3 and 0.5 microg Cd g(-1) dry weight) except for male values in October (mean 1 microg Cd g(-1) dry weight). Some of the seasonal changes in cadmium content of the crabs might plausibly be explained by changes in cadmium uptake from water, i.e. changes during the moult cycle and changes in cadmium uptake rates from water brought about by changes in ambient factors such as salinity and temperature. However, uptake of cadmium from water and transfer to the midgut gland take place at a rate that is two orders of magnitude too low to account for the increase in the cadmium concentrations in midgut gland in male crabs between May and June. The distribution of cadmium among tissues in crabs collected at uncontaminated sites also corresponds better with results obtained after administration of cadmium via the food than via water, and the exposure of the crabs to cadmium via the food is large enough to explain the increase in concentration between May and June. PMID- 15748745 TI - Association between cadmium and calcium uptake and distribution during the moult cycle of female shore crabs, Carcinus maenas: an in vivo study. AB - Net influxes into the haemolymph and tissue distribution of 45Ca and 109Cd were studied in vivo in female Carcinus maenas at different moult stages. Net influxes of 45Ca and 109Cd from water were higher in postmoult (A and B) C. maenas than in C3- and C4-intermoult crabs and the net influx of calcium was higher in C3 intermoult crabs than in C4-intermoult crabs. The net influxes of 45Ca and 109Cd increased in postmoult C. maenas with decreasing external calcium concentrations at constant salinity. At all external calcium concentrations a significant correlation existed between 45Ca and 109Cd accumulated in the haemolymph of individual animals. In vivo exposure of postmoult C. maenas to external lanthanum decreased the 45Ca and 109Cd uptake rates to 30 and 10%, respectively, of the control values. About 30% of injected 109Cd were found in the midgut gland, 10 20% in the gills and only a few (1-2) percent was lost to the seawater 24 h after injection. No major variations in tissue distribution of 109Cd were observed between moult stages in these tissues. Premoult crabs retained more cadmium in the haemolymph 24 h after injection than other moult stages, and postmoult crabs retained more in muscle. Between 20 and 40% of the injected 45Ca were excreted to the water, while only a few percent of the injected 45Ca were found in the soft tissues 24 h after injection. Large moult stage variations, however, were observed in the tissue distribution of internalised 45Ca. This study demonstrates that cadmium and calcium uptakes are elevated in postmoult C. maenas. The results indicate that cadmium and calcium in this stage are taken up via Ca2+-channels located in the apical membrane of gill epithelium cells. When internalised, however, cadmium and calcium are metabolised in fundamentally different ways, determined by the chemical properties and biological significance of the two metals. PMID- 15748746 TI - In vivo and in vitro cadmium accumulation during the moult cycle of the male shore crab Carcinus maenas--interaction with calcium metabolism. AB - The effect of moult stage on cadmium accumulation and distribution was investigated in vivo in male shore crabs Carcinus maenas exposed to 1 mg Cd l(-1) for 7 days. The accumulation of cadmium in all tissues examined was markedly higher in postmoult (A(1-2) and B(1-2)) compared to intermoult (C1, C3 and C4) and premoult (D(0-3)). In addition, elevated levels of cadmium were found in gills of late premoult (D(2-3)) animals. The total amount of cadmium accumulated in the tissues (haemolymph, gills, midgut gland and muscle) increased from 43 microg Cd in early premoult (D(0-1)) to 391 microg Cd in late postmoult (B(1-2)). Gills and midgut gland were the primary cadmium accumulating tissues in C4 intermoult and premoult (D(0-3)); in early postmoult (A(1-2)) haemolymph and midgut gland were the main cadmium containing tissues, while midgut gland dominated in late postmoult (B(1-2)) and early intermoult (C1 and C3). A detailed account of calcium distribution in haemolymph, gills, midgut gland, muscle and exoskeleton during the moult cycle is presented. Mechanistic links between cadmium and calcium uptake in posterior gills of C4-intermoult and early postmoult (A(1-2)) crabs were explored using an in vitro gill perfusion technique. Calcium and cadmium influxes were markedly higher in postmoult compared to intermoult. No differences between intermoult and postmoult effluxes were found for either calcium or cadmium. From intermoult to postmoult net influx increased from 2.4 to 29 micromol Ca2+ g(-1) ww(gill) h(-1) and from 0.24 to 25 nmol Cd2+ g(-1) ww(gill) h(-1). The results indicate that the postmoult increase in cadmium influx is due to increased active transport of cadmium, at least partly, by accidental uptake via calcium transporting proteins. The in vitro net influx rates corresponded accurately to the observed in vivo accumulation of both cadmium and calcium. Although cadmium accumulation and distribution are clearly linked to changes in calcium requirements, cadmium did not interfere with calcium accumulation or distribution at any stage during the moult cycle. PMID- 15748747 TI - Cadmium, zinc and the uptake of calcium by two crabs, Carcinus maenas and Eriocheir sinensis. AB - The uptake of dissolved cadmium and zinc by crustaceans can usually be explained by the passive process of facilitated diffusion involving a transport protein in the membranes of permeable surfaces. Cadmium ions will also enter via uptake routes for calcium, given the similar size of the two free ions. This study has investigated the interaction of cadmium (and comparatively zinc) and calcium uptake in two crabs that show different permeability responses to changes in salinity, with consequently different effects on the uptake of cadmium and zinc with salinity change. Ca uptake rates in Carcinus maenas decreased in reduced salinity (33-15) with the decreased Ca concentration of the medium and increased if the Ca concentration was increased at salinity 20. It is concluded that Ca uptake over the salinity range 33-15 is via apical Ca channels in gill ionocytes, passively down an electrochemical gradient. The Ca uptake rate of Eriocheir sinensis showed no significant decrease over the salinity range 33-10 (probably because of the small differences in an already low Ca uptake rate in this crab against a background of inter-individual variability), but decreased significantly at salinity 5. Added calcium increased the Ca uptake rate of E. sinensis at salinities 15 and 5, supporting the interpretation that Ca uptake in gills is typically passive via apical Ca channels. Cadmium (but not zinc) inhibited calcium uptake in both crabs at 15 salinity, indicating sharing of Ca channels by Cd, but not at salinity 5 (E. sinensis only) when Ca may be taken up into gill ionocytes by another (active?) physiological process. PMID- 15748748 TI - Mechanism of acute silver toxicity in marine invertebrates. AB - In freshwater crustaceans and in both freshwater and marine fish, the key mechanism of acute silver toxicity involves ionoregulatory impairment. An inhibition of the Na+ ,K+-ATPase located at the basolateral membrane of the gill epithelium seems to be the key site for silver toxicity. However, studies to determine if the same mechanism of toxicity is occurring in marine invertebrates, which also are ionoregulators, had not been done. Thus, the present study was carried out to determine acute silver effects on hemolymph osmo- and ionoregulation in three marine invertebrates: the shrimp Penaeus duorarum, the sea hare Aplysia californica, and the sea urchin Diadema antillarum. Animals were exposed to silver (1 or 10 microg/L), as silver nitrate, in seawater for 48 h. Results show that acute silver exposure did not affect hemolymph osmolality or ion concentration (Na+, Cl-, K+, Ca2+ and Mg2+) in the three species studied. However, silver induced significant changes in the water content in shrimp gill and sea hare gill and hepatopancreas. Silver also caused significant changes in Na+ ,K+-ATPase activity and in both total and intracellular ion (Cl-, Na+, K+, Mg2+, and Ca2+) concentrations in different tissues of the three species studied. Overall, these results show that the key mechanism of acute silver toxicity in marine invertebrates is not associated with an osmotic or ionoregulatory impairment at the hemolymph level, as observed in freshwater fish and crustaceans and in seawater fish. However, they indicate that acute waterborne silver induces significant changes in Na+ ,K(+)-ATPase activity and probably affects other mechanisms involved in water and ion transport at the cell membrane level, inducing impairments in water and ion regulation at the cellular level in different tissues of marine invertebrates. These results indicate the need to consider other "toxic sites" than gills in any future extension of the biotic ligand model (BLM) for seawater. PMID- 15748749 TI - Cadmium-handling strategies in two chronically exposed indigenous freshwater organisms--the yellow perch (Perca flavescens) and the floater mollusc (Pyganodon grandis). AB - Laboratory experiments on a variety of aquatic organisms suggest that metallothionein-like proteins (MT) play an important role in the regulation of essential metals, and in the sequestration and detoxification of non-essential metals (e.g., Cd). However, the importance of metallothionein production relative to alternative strategies of metal detoxification, and its effectiveness in metal detoxification, remain largely unexplored in field situations. In the present study we explored metal-handling strategies in an adult benthic bivalve (Pyganodon grandis) and in juvenile yellow perch (Perca flavescens), exposed to Cd in their natural habitat. The two biomonitor species were collected from lakes located along a Cd concentration gradient. Ambient dissolved Cd concentrations were determined by in situ dialysis as a measure of metal exposure. Sub-cellular Cd partitioning was determined in target tissues (bivalve gills and digestive gland; perch liver) by differential centrifugation, and metallothionein was measured independently by a mercury-saturation assay in the bivalve tissues. Malondialdehyde concentrations were measured as a potential indicator of oxidative stress. Ambient dissolved Cd concentrations ranged from 0.06 to 0.57 nM in the nine lakes from which bivalves were collected, and from < 0.3 to 6.7 nM in the eight lakes from which yellow perch were sampled. Bioaccumulated Cd also varied from lake to lake, more so for the bivalve than for the yellow perch; the [Cd]max/[Cd]min ratios for the various tissues decreased in the order: bivalve gill Cd 28 > bivalve digestive gland Cd 18 > perch hepatic Cd 14. In the two lakes that were common to both the bivalve and perch studies, i.e. lakes Opasatica and Vaudray, accumulated Cd concentrations were consistently higher in the bivalve than in the perch. Cadmium-handling strategies were similar in the bivalve digestive gland and perch liver, in that Cd was mainly associated with the heat-stable protein (HSP) fraction. Furthermore, in these organs the contributions from the "mitochondria" and "lysosomes + microsomes" fractions were consistently higher than in the gill tissue. In the bivalve gill, the HSP fraction could only account for a small proportion (10+/-3%) of the total Cd burden, and the metal was instead largely sequestered in calcium concretions (58+/-13%). Along the Cd-exposure gradient, Cd detoxification appeared to be reasonably effective in the bivalve gill and digestive gland, as judged from the protection of the heat-denaturable protein (HDP) fraction. However, in both organs Cd concentrations did increase in potentially metal-sensitive organelles (mitochondria), and malondialdehyde concentrations increased along the exposure gradient in the gills (but not in the digestive gland). Cadmium detoxification seemed less effective in juvenile yellow perch. As total hepatic Cd increased, Cd concentrations increased in all sub-cellular fractions, including the HDP fraction that was well protected in the bivalve. The relative proportions of Cd in the various fractions did not vary appreciably along the exposure gradient and there was no evidence of a threshold exposure concentration below which sensitive metal pools were protected. PMID- 15748750 TI - Effects of dietary calcium and cadmium on cadmium accumulation, calcium and cadmium uptake from the water, and their interactions in juvenile rainbow trout. AB - The objective of this study was to examine the effects of chronically elevated dietary Ca2+ (as CaCO3), alone and in combination with elevated dietary Cd, on survival, growth, and Cd and Ca2+ accumulation in several internal compartments in juvenile rainbow trout (Oncorhynchus mykiss). In addition, effects on short term branchial uptake and internal distribution of newly accumulated waterborne Ca2+ and Cd during acute waterborne Cd exposure (50 microg/L as CdNO3 for 3 h) were monitored using radiotracers (45Ca, 65Cd). Fish were fed with four diets: 20 mg Ca2+/g food (control), 50 mg Ca2+/g food, 300 microg Cd/g food, and 50 mg Ca2+/g + 300 microg Cd/g food for 30 days. There were no significant effects on growth, mortality, or total body Ca2+ accumulation. The presence of elevated Ca2+, Cd, or Ca2+ + Cd in the diet all reduced waterborne Ca2+ uptake in a short term experiment (3 h), though the inhibitory mechanisms appeared to differ. The effects were marked after 15 days of feeding, but attenuated by 30 days, except when the diet was elevated in both Ca2+ and Cd. The presence of elevated Ca2+ in the diet had only modest influence on Cd uptake from the water during acute Cd challenges but greatly depressed Cd uptake from the diet and accumulation in most internal tissues. None of the treatment diets prevented the decreases in waterborne Ca2+ uptake and new Ca2+ accumulation in internal tissues caused by acute exposure to waterborne Cd. In conclusion, there are complex interactions between waterborne and dietary effects of Ca2+ and Cd. Elevated dietary Ca2+ protects against both dietary and waterborne Cd uptake, whereas both waterborne and dietary Cd elevations cause reduced waterborne Ca2+ uptake. PMID- 15748751 TI - Renal function in the freshwater rainbow trout (Oncorhynchus mykiss) following acute and prolonged exposure to waterborne nickel. AB - Renal function was investigated in adult rainbow trout following acute and prolonged exposure to waterborne Ni in moderately hard Lake Ontario water (approximately 140 mgL(-1) as CaCO3). Fish were exposed for 36 days to a sublethal concentration of 442 microg Ni L(-1), followed by 96 h of exposure to 12,850 microg Ni L(-1) (approximately 33% of the 96 h LC50). Prolonged exposure markedly affected only the renal handling of Ni, with no substantial effect on the plasma concentration, urinary excretion rate (UER) or clearance ratio (CR) of Na+, Cl-, K+, Ca2+, Mg2+, inorganic phosphate (P(i)), glucose, lactate, total ammonia (T(amm)), protein and free amino acids (FAA). Glomerular filtration rate (GFR) was reduced by 75% over 96 h of acute Ni challenge in both fish previously exposed to Ni and naive fish, with no significant change in urine flow rate (UFR), suggesting a substantial reduction in water reabsorption to maintain urine flow and water balance. Renal Mg2+ handling was specifically impaired by acute Ni challenge, leading to a significantly increased UER(Mg2+) and significantly decreased plasma [Mg2+] only in naive fish. Previously-exposed fish were well protected against Ni-induced Mg2+ antagonism, indicating true acclimation to Ni. Only in naive, acutely challenged fish was there an increased UER of titratable acidity (TA-HCO3), net acidic equivalents, P(i), T(amm) and K+. Again, all of these parameters were well-conserved in previously-exposed fish during acute Ni exposure, strongly suggesting that prolonged, sublethal exposure protected against acute Ni-induced respiratory toxicity. PMID- 15748752 TI - The changes to apical silver membrane uptake, and basolateral membrane silver export in the gills of rainbow trout (Oncorhynchus mykiss) on exposure to sublethal silver concentrations. AB - Juvenile rainbow trout acclimated to softwater were exposed to 0 or 8.3 nM Ag (added as silver nitrate) for 21 days. On days 1, 7 and 21 gill, kidney and liver levels of silver; branchial Na+ influx, efflux and net flux rate; gill and kidney K+ -dependent p-nitrophenol phosphatase activity; and gill and liver accumulation of "new" Ag were measured. In addition, the concentration-dependent uptake of Ag by gill basolateral membrane vesicles (BLMV) was assessed in control fish and those exposed to 8.3 nM Ag for 7 days. Ag induced a significant increase in Na+ efflux following 1 day of exposure that resulted in an increase in net loss of Na+ and a reduction in Na+ influx. By day 21 this perturbation to Na+ balance had been corrected, but kidney K+ -dependent p-nitrophenol phosphatase activity was significantly reduced. Unexpectedly, the Ag concentrations in the liver of Ag exposed fish only significantly increased (two-fold) following 7 days of exposure and were not elevated when compared to controls on day 21. In contrast, the gill and kidney accumulated significant concentrations of Ag (20-fold increase) following 7 days of exposure, and the Ag concentration in these tissues remained similar on day 21. The gills of Ag exposed fish accumulated significantly less "new" Ag than the controls on days 7 and 21 following exposure, suggesting a down regulation of branchial Ag uptake. The BLMV of Ag exposed fish showed a significant increase in V(max) [control fish BLMV V(max) = 2811.9+/-190.8 pmol (110 m)Ag/(mg protein x min) and Ag exposed fish BLMV V(max) = 3688.3+/-659.8 pmol (110 m)Ag/(mg protein x min) (P = 0.033)], suggesting that they are able to increase export of Ag from the gills on exposure to Ag. The results from this study demonstrate a complex array of physiological processes that control the bioreactive concentrations of Ag in the gills, including: cytoplasmic sequestration, a down-regulation of apical entry and potentially an increase in basolateral membrane extrusion. PMID- 15748753 TI - Dose-dependent inorganic mercury absorption by isolated perfused intestine of rainbow trout, Oncorhynchus mykiss, involves both amiloride-sensitive and energy dependent pathways. AB - The trophic transfer and nutritional toxicity of mercury (Hg) in aquatic food chains is well known, but there is limited information on the mechanism of mercury uptake across the gut. In this study, isolated whole gut sacs from rainbow trout were used to identify the regions of the gut involved in Hg absorption, and then perfused intestines were used to investigate Hg uptake. Exposure of whole gut sacs to 100 micromol l(-1) Hg as HgCl2 in the luminal solution caused Hg accumulation primarily in the mucosa (78% or more), with the intact mid and hind gut supporting 59% of the accumulated Hg. Luminal exposure to [Hg] between 0 and 100 micromol l(-1) for 4 h in perfused trout intestines showed a non-linear dose-dependent accumulation with a maximum Hg uptake rate of about 103 nmol g(-1) h(-1), and suggests carrier mediated transport into the gut cells and the blood. Additions of 2 mmol l(-1) amiloride depressed Hg accumulation by the mid and hind gut by 40-50%, whilst additions of the Ca chelator 1 mmol l(-1) EGTA increased Hg levels in the tissue. Symmetrical additions of 10 mmol l(-1) cyanide did not prevent tissue accumulation of Hg, but caused a 3.4-fold decline in net Hg flux to the serosal compartment. We conclude that Hg absorption across the gut is partly carrier mediated and involves both amiloride sensitive, and energy-dependent pathways. PMID- 15748754 TI - Influence of water chemistry and natural organic matter on active and passive uptake of inorganic mercury by gills of rainbow trout (Oncorhynchus mykiss). AB - To distinguish physiologically regulated uptake from passive uptake of inorganic Hg in fish, rainbow trout (Oncorhynchus mykiss) were exposed to inorganic Hg (0.5, 1, or 2 microM total Hg) in ion-poor water with various treatments. Addition of ions to the water (mM concentrations of Ca, K, Cl) did not consistently alter Hg accumulation by trout gills, although there was a trend to higher Hg accumulation at higher ion concentrations. The apical Ca channel blockers Verapamil and lanthanum also did not consistently affect Hg accumulation by trout gills. Pre-treatment of trout with the Na channel blocker Phenamil decreased Hg uptake by about half. These results suggest a combination of physiologically regulated and passive uptake of Hg by trout gills. Strong complexing agents of Hg (EDTA, NTA, ethylenediamine, cysteine) decreased Hg binding by trout gills in a dose-dependent manner. From these data, a conditional equilibrium binding constant for Hg to the gills was estimated as logK(Hg-gill) = 18.0, representing very strong binding of Hg to the gills. This value is a first step in creating a biotic ligand model (BLM) for inorganic Hg and fish. Natural organic matter (2-10 mg C/L) also decreased Hg-binding by trout gills, although mM concentrations of Na, K, and Cl interfered with this effect. At low concentrations of these ions, natural organic matter samples isolated from various sources bound Hg to similar degrees, as judged by Hg accumulation by trout gills. A conditional binding constant to natural organic matter (NOM) was estimated as logK(Hg-NOM) = 18.0 with about 0.5 micromol binding sites per mg C, representing strong binding of Hg to NOM. PMID- 15748756 TI - Tuberculosis and brucellosis prevalence survey on dairy cattle in Mbarara milk basin (Uganda). AB - We determined the prevalence of tuberculosis and brucellosis reactors in the dairy herds in the Mbarara district of Uganda in 2002. This is one of the most important dairy-production areas of the country and includes both pastoral and agro-pastoral zones. A total of 340 (of 11,995) randomly selected herds were tested for tuberculosis, using the intradermal tuberculosis-skin test and 315 (of 10,562) herds tested for brucellosis using the serum Rose Bengal test. The herd prevalence for tuberculosis reactors was 74.1% (95% confidence intervals 69, 78), the individual-animal prevalence was of 6.0% (5.6, 6.5) and within-herd range was 1-50% (up to 100% if suspicious reactors were included). The herd prevalence for brucellosis was 55.6% (50, 61.2) individual-animal prevalence 15.8% (14.8, 16.7) and within-herd range 1-90%. The reactor prevalence increased with the age of the animals for both tuberculosis and brucellosis. Tuberculosis reactor prevalences were higher in animals from the agro-pastoral zone. However, the individual animal and herd prevalences of brucellosis seroprevalences were higher in the pastoral zone. PMID- 15748755 TI - The impact of badger removal on the control of tuberculosis in cattle herds in Ireland. AB - In Ireland, the herd prevalence of bovine tuberculosis has remained stable for several decades, and in common with several other countries, progress towards eradication has stalled. There is evidence in support of the potential role of infected badgers (Meles meles, a protected species) in bovine tuberculosis in Ireland and Britain. However, this evidence on its own has not been sufficient to prove disease causation. Field trials are likely to offer the best opportunity to define this role. Building on the earlier East Offaly project, our objectives were to assess the impact of badger removal on the control of tuberculosis in cattle herds in Ireland. The study was conducted from September 1997 to August 2002 in matched removal and reference areas (average area of 245.1km(2)) in four counties: Cork, Donegal, Kilkenny and Monaghan. Badger removal was intensive and proactive throughout the study period in the removal areas, but reactive (in response to severe tuberculosis outbreaks in cattle) in the reference areas. Removal intensity in the removal and reference areas during the first 2 years of the study averaged 0.57 and 0.07 badgers/km(2)/year, respectively. The outcome of interest was restriction of cattle herds due to confirmed tuberculosis, where tuberculous lesions were detected in one or more animals. Data were analysed using logistic regression (modelling the probability of a confirmed herd restriction) and survival analysis (modelling time to a confirmed herd restriction). During the study period, there was a significant difference between the removal and reference areas in all four counties in both the probability of and the time to a confirmed herd restriction due to tuberculosis. In the final year of the study, the odds of a confirmed herd restriction in the removal (as compared to the reference areas) were 0.25 in Cork, 0.04 in Donegal, 0.26 in Kilkenny and 0.43 in Monaghan. Further, the hazard ratios (removal over reference) ranged from 0.4 to 0.04 (a 60-96% decrease in the rate at which herds were becoming the subject of a confirmed restriction). PMID- 15748757 TI - Evaluation of surveillance strategies for bovine tuberculosis (Mycobacterium bovis) using an individual based epidemiological model. AB - The Netherlands holds the bovine tuberculosis-free (BTB-free) status according to European Union standards, but in recent years small outbreaks of the infection have occurred. After the last outbreak in 1999 with 10 infected herds the question raised if the current surveillance system, visual inspection of carcasses at the slaughterhouse, is efficient enough to detect infected cattle in time and to maintain the official BTB-free status. Through epidemiological modelling, the risk of a major outbreak is quantified, using one of six surveillance strategies. These are the currently used visual inspection of carcasses at the slaughterhouse (SL), the ELISA test on blood samples of carcasses at the slaughterhouse (ELISA-B), the gamma-interferon test on blood samples of carcasses at the slaughterhouse (GAMMA-B), comparative tuberculination of the herd (CT), the combined method of single and comparative tuberculination of the herd (ST+CT) and the ELISA test on samples of bulk milk (ELISA-M). Test frequency of the last three methods was varied as well. A stochastic individual based model (IBM) was developed to simulate a chain of infected herds, where each individual animal is followed in time. The model mimics the nation-wide situation after the introduction of one infected animal into one herd. BTB-transmission is simulated with an S-E(1)-E(2)-I state transition model. Output is time until detection of the infection, prevalence in the detected herd and the number of infected herds at the time of detection. For the assessment 500 simulations were used, representing 500 BTB-introductions. Model robustness to parameter values was analysed with Monte Carlo elasticity analysis, for which 1000 simulations were used. Results of median time until detection and median number of infected farms at detection for SL (302 weeks and seven farms) were in agreement with estimates from an outbreak in the Netherlands in 1999. ELISA-B and GAMMA-B performed better than SL with a much lower median time until detection (189 and 97 weeks, respectively). The results for the tuberculination methods (ST+CT and CT) and ELISA-M depended heavily on the frequency in which the tests were performed. The tuberculination methods ST+CT and CT yield comparable results and detect the infection sooner than SL, also at the lowest tested frequency of once in 5 years. ELISA-M is comparable with SL at frequencies of once in 4 or 5 years, and this test works well at frequencies of once a year or higher. Our study results are used for an economical optimisation analysis of the six surveillance strategies. PMID- 15748758 TI - A case study of capture-recapture methodology using scrapie surveillance data in Great Britain. AB - We applied capture-recapture methodology (CRC) to data from three surveillance sources (statutory notification, abattoir survey (AS) and fallen stock (FS) survey) to estimate the number of holdings infected with scrapie in Great Britain and to assess the sensitivity of the surveillance network. Between January 1, 2002 and March 31, 2003, 144 holdings were identified by the three sources. Using CRC modelling techniques, we estimated a minimum lower bound for the total number of holdings infected as 642. A biologically plausible positive dependence between the statutory reporting and the fallen stock survey was found statistically significant. The sensitivity of the three sources combined was very low. The integration of the three overlapping sources provided a better understanding of the interactions within the surveillance network. However, the scarcity of the data and reduced overlapping among sources only allowed for very cautious inferences to be drawn about the true proportion of scrapie affected holdings in the national population. Future surveys and surveillance activities should be planned such that the resulting data can be used more effectively as part of CRC modelling approaches. PMID- 15748759 TI - Combined effect of the methanol utilization (Mut) phenotype and gene dosage on recombinant protein production in Pichia pastoris fed-batch cultures. AB - An important number of heterologous proteins have been produced in the methylotrophic yeast Pichia pastoris using the alcohol oxidase promoter. Two factors that drastically influence protein production and cultivation process development in this system are gene dosage and methanol assimilation capacity of the host strain (Mut phenotype). Using a battery of four strains which secrete a Rhizopus oryzae lipase (ROL), the combined effects of gene dosage and Mut phenotype on recombinant protein production in Pichia pastoris was studied in fed batch cultures. Regarding the effect of phenotype, the specific productivity and the Y(P/X) were 1.29- and 2.34-fold higher for Mut(s)ROL single copy strain than for Mut+ROL single copy strain. On the contrary, the productivity of Mut+ROL single copy strain was 1.34-fold higher than Mut(s)ROL single copy strain. An increase in ROL gene dosage seems to negatively affect cell's performance in bioreactor cultures, particularly in Mut(s) strains. Overall, the Mut(s) strain may be still advantageous to use because it allows for easier process control strategies. PMID- 15748760 TI - Three forms of thermostable lactose-hydrolase from Thermus sp. IB-21: cloning, expression, and enzyme characterization. AB - Three thermostable lactose-hydrolases, namely, two beta-glycosidases (bglA and bglB) and one beta-galactosidase (bgaA) genes were cloned from the genomic library of Thermus sp. IB-21. The bglA, bglB, and bgaA consisted of 1311 bp (436 amino acid residues), 1296 bp (431 aa), and 1938 bp (645 aa) of nucleotides with predicted molecular masses of 49,066, 48,679, and 72,714 Da, respectively. These enzymes were overexpressed in Escherichia coli BL21(DE3) using pET21b(+) vector system. The recombinant enzymes were purified to homogeneity by a heat precipitation (70 degrees C, 40 min) and a Ni2+-affinity chromatography. The molecular masses of the purified enzymes estimated by SDS-PAGE agreed with their predicted values. All the purified enzymes showed their optimal pH at around 5.0 6.0. In contrast, the temperature profiles for activity and thermostability patterns were different for each enzyme. BglB beta-glycosidase displayed the best lactose hydrolysis activity of the three enzymes without substrate inhibition up to 200 mM lactose at 70 degrees C and pH 7.0. The specific activities (U/mg) of BglA, BglB, and BgaA on 138 mM lactose at 70 degrees C and pH 7.0 were 36.8, 160.3, and 8.5, respectively. PMID- 15748761 TI - Investigations of the transfructosylation reaction by fructosyltransferase from B. subtilis NCIMB 11871 for the synthesis of the sucrose analogue galactosyl fructoside. AB - The exo-fructosyltransferase produced from B. subtilis NCIMB 11871 strain transfers the fructose moiety from donor alpha12 linked saccharides such as sucrose, raffinose and stachyose to the acceptor d-galactose, leading to the sucrose analogue, galactosyl-fructoside. Here, we report detailed kinetic studies. The enzyme showed a remarkably high optimal temperature at 50 degrees C and was effectively immobilised on Eupergit C 250 L and Trisopor-Amino. This is also the first report about the equilibrium of the transfructosylation reaction, its activation energy determination, the structure of the product and its preparative scale isolation. PMID- 15748762 TI - Enhanced activity of recombinant beta-secretase from Drosophila melanogaster S2 cells transformed with cDNAs encoding human beta1,4-galactosyltransferase and Galbeta1,4-GlcNAc alpha2,6-sialyltransferase. AB - beta-Secretase (betaSEC) was expressed in Drososphila melanogaster Schneider 2 (S2) cells transformed with cDNAs encoding beta1,4-galactosyltransferase (GalT) and Galbeta1,4-GlcNAc alpha2,6-sialyltransferase (ST). The apparent molecular weight of recombinant beta-secretase was increased from 56kDa to 61kDa. A lectin blot analysis indicated that recombinant beta-secretase from S2betaSEC/GalT-ST cells (S2 cells co-transformed with cDNAs encoding beta-secretase, glycosyltransferases, GalT, and ST) contained the glycan residues of beta1,4 linked galactose and alpha2,6-linked sialic acid. Two dimensional electrophoresis revealed that recombinant beta-secretase from S2betaSEC/GalT-ST cells had a lower isoelectric point compared to beta-secretase from control S2betaSEC cells (S2 cells transformed only with beta-secretase cDNA). Recombinant beta-secretase from transformed S2 cells was also present as heterogeneous forms. The enzyme activity of recombinant beta-secretase from S2betaSEC/GalT-ST cells was enhanced up to 260% compared to control S2betaSEC cells. We have shown that an exogeneous human glycosyltransferases cDNA can be introduced into S2 cells to extend the N-glycan processing capabilities of the insect cell line, and that the extended glycosylation improves the activity of recombinant beta-secretase. PMID- 15748763 TI - Application of Aspergillus niger-treated agrowaste residue and Glomus mosseae for improving growth and nutrition of Trifolium repens in a Cd-contaminated soil. AB - The microbial transformation of sugar beet (SB) agrowaste with or without rock phosphate (RP) has utility for the improvement of plant growth in a Cd (5 microg g-1) artificially contaminated soil, particularly when the soil is co-inoculated with arbuscular mycorrhizal (AM) fungus Glomus mosseae isolated from a Cd polluted area. Under such Cd-polluted conditions, the limited growth, mineral nutrition, symbiotic developments (nodulation and AM-colonization) and soil enzymatic activities were stimulated using SB or SB+RP as soil amendments and G. mosseae as inoculant. G. mosseae enhanced plant establishment in a higher extent in amended soil; it is probably due to the interactive effect increasing the potential fertility of such compounds and its ability for decreasing Cd transfer from soil to plant. The amount of Cd transferred from soil solution to biomass of AM-colonized plants ranged from 0.09 microg Cd g-1 (in SB+RP-amended soil) to 0.6 microg Cd g-1 (in non-amended soil). Nodule formation was more sensitive to Cd than AM-colonization, and both symbioses were stimulated in amended soils. Not only AM-colonization but also amendments were critical for plant growth and nutrition in Cd-polluted soil. The high effectiveness of AM inoculum increasing nutrients and decreasing Cd in amended soil indicated the positive interaction of these treatments in increasing plant tolerance to Cd contamination. PMID- 15748764 TI - Transglutaminase treatment of wool fabrics leads to resistance to detergent damage. AB - Biological detergents are now routinely used in domestic laundry because the enzymes they contain provide the added benefit of low temperature washes with improved cleaning performance. One of the key enzymes found in these detergents are proteases, which if exposed to natural protein fibres such as wool or silk can cause irreversible damage, leading to loss of fabric strength, shape and poor colour fastness. Transglutaminases (TGases) are protein cross-linking enzymes capable of adding tensile strength to wool proteins, and as a consequence are capable of remediating the damage caused by previous chemical treatments, and more importantly, by proteases. In this paper we treated dyed wool fabric with TGase and then washed the fabric with biological and non-biological detergents to investigate whether TGases would protect wool garments from damage by the undue use of biological detergents in domestic laundry. We demonstrate using different cycles of detergent washes containing biological and non-biological detergents and different TGase treatments, that wool fabric treated previously with TGase release less dye into the washing liquor and in addition maintain fabric strength at levels greater than the washed controls. As a consequence, wool garments previously treated with TGase are likely to have increased resistance to domestic washing and thus provide increased longevity. PMID- 15748765 TI - Gas holdup, foaming and oxygen transfer in a jet loop bioreactor with artificial foaming media and yeast culture. AB - A concentric draft tube jet loop bioreactor (10.5 m3) was used to study the influence of aerated liquid height (above the draft tube) on the amount of surfactant addition allowable without foaming. Sodium lauryl sulfate (SLS) and defatted soybean flour in tap water were used as model artificial media. The amount of surfactant required to develop foaming and the maximum gas holdup achieved prior to foaming were notably influenced by aerated liquid height. Decreasing the aerated liquid height from 1.50 to 0.05 m increased the amount of SLS allowed without foaming from 2.2 to 12.1 g, the gas holdup in the riser from 0.18 to 0.31 and the gas holdup in the downcomer from 0.12 to 0.25. Similar behavior was observed for soybean flour. Decreasing the aerated liquid height from 1.45 to 0.05 m increased the amount of soybean flour allowed without foaming from 822 to 3200 g, the gas holdup in the riser from 0.17 to 0.26 and the gas holdup in the downcomer from 0.10 to 0.19. Data from a representative continuous yeast culture are reported to show that operation at low aerated liquid heights (0.5 m) can also be used to produce a culture broth with large gas holdup and oxygen transfer but without foaming. PMID- 15748766 TI - Immobilization of Candida rugosa lipase on a pH-sensitive support for enantioselective hydrolysis of ketoprofen ester. AB - Candida rugosa lipase (Lipase OF) was immobilized by covalent binding to a pH sensitive support showing reversibly soluble-insoluble characteristics with pH change. The immobilized lipase could carry out the enantioselective hydrolysis of ketoprofen ester in a soluble form yet be recovered after precipitation by simply adjusting pH. Its activity and enantioselectivity for hydrolysis of 2-chloroethyl ester of ketoprofen were enhanced 1.5-fold and 8.7-fold compared with those of free lipase. After eight catalytic cycles, the immobilized enzyme was still 46% active and its enantioselectivity remained unchanged. PMID- 15748767 TI - Evaluation of a new annular capacitance probe for biomass monitoring in industrial pilot-scale fermentations. AB - The four-pin electrode capacitance probe has already shown to be a valuable tool for on-line monitoring viable biomass concentration in industrial-type fermentations. A new prototype annular probe was developed and its performance in real-time monitoring the concentration of viable cells during industrial pilot scale fermentation for the production of an Active Pharmaceutical Ingredient (API) was investigated and compared to the four-pin probe. A set of 14 fermentations was monitored on-line: four of them with the four-pin probe, the remaining with the annular probe. The performance of both the annular and the four-pin electrode probe were compared against each other and against off-line measurements (viscosity and packed mycelial volume). The prototype annular probe showed to have higher signal intensity and sensitivity than the standard four-pin probe, with higher signal-to-noise ratio. Furthermore, its new design and construction proved to be easier to handle in an industrial environment. PMID- 15748768 TI - Liquid-liquid extraction of commercial glucose oxidase by reversed micelles. AB - Aim of this work was to establish the optimum operating conditions for the extraction and recovery by cationic reversed micelles of commercial glucose oxidase (GOX) from Aspergillus niger, in view of possible application to raw cell homogenates. The influence of pH, temperature, electric conductivity and solvent/co-solvents ratio on the extraction was investigated by a fractional factorial design of 2(3-1) type, conjugated with a mixture experimental design, using the residual enzyme activity to evaluate the results. The best conditions for GOX extraction were ensured using isooctane as solvent and hexanol and butanol as co-solvents at 76/6/18 volume ratio, pH 6.0, 0.2 M cetyl trimethylammonium bromide (CTAB) as cationic surfactant, and electrical conductivity (kappa) of 4.8 mS cm-1. The highest yield of GOX activity recovery (about 90%) was in fair accordance with the value predicted by the model. PMID- 15748769 TI - [Evaluation after four years of exercise therapy for chronic low back pain]. AB - OBJECTIVE: To assess at four years follow-up the efficiency of exercise therapy for chronic low back pain in terms of pain, physical ability, quality of life and return to work. MATERIALS AND METHODS: Patients who graduated from a functional restoration program between April 1997 and June 1999, answered a questionnaire at one year follow-up. The patients who had answered this questionnaire were evaluated 48 months later: they first answered another questionnaire and then most underwent a clinical examination assessing pain on a visual analogue scale, flexibility by use of the Schober index and the finger-ground distance test, endurance of abdominal and spinal muscles, and quality of life as assessed by the Dallas pain questionnaire. RESULTS: Thirty-four patients graduated from the program; 26 were studied at four-year follow-up. Pain intensity was significantly reduced. Improvement in flexibility after the program remained at four-year follow-up, whereas improvement in endurance did not. Scores on the Dallas pain questionnaire remained as improved as those at one-year follow-up. Initially, 23 of the 26 patients had a job; 19 were on sick leave for an average of 35 weeks. At four-year follow-up, 16 patients were still working, and 56% had changed jobs. Six patients pursued a regular activity before entering the program; at four-year follow-up, 17 were regularly active. No significant relationship was found between the pursuit of physical activity and return to work. Nevertheless, a significant relationship was found between having a regular physical activity and improvement of the finger-ground distance and endurance of spinal muscles. This observation was not true for the Dallas pain questionnaire scores. CONCLUSION: Despite the lack of a control group, this study seems to favour a benefit in the functional restoration program. PMID- 15748770 TI - [The neurobehavioral rating scale-revised: assessment of concurrent validity]. AB - Cognitive and behavioral impairment are a major source of disability in daily living of patients with traumatic brain injury (TBI). The Neurobehavioral Rating Scale-Revised (NRS-R) is a short, easy-to administer interview tool developed to improve assessment by clinicians. Data are available on its criterion validity and reliability, but the concurrent validity of the French NRS-R was not yet documented. OBJECTIVE: To assess the concurrent validity of the NRS-R with current psychometric tests. METHOD: One hundred and four patients with TBI enrolled in a community adjustment program underwent concurrent examination with the NRS-R, cognitive tests assessing memory, attention, and executive functions, and scales of anxiety (STAI) and depressive mood (CES-D). Intercorrelations were undertaken between these variables and the five factors of the NRS-R: F1, intentional behavior; F2, lowered emotional state; F3, survival-oriented behaviour/hightened emotional state; F4, arousal state; and F5, language. Patients were 82 men and 22 women, the mean age was 28.5 years, and 70% had severe TBI (Glasgow coma score [GCS] below 8 on admission). They were assessed 52 months on average after their injury. RESULTS: Factor F1 was correlated with results on the GCS (P<0.05), the Tower of London test (TL, P<0.01), the Trail Making Test (TMT, P<0.01), divided attention (DA) and inhibition (IN) subscales of the Zimmermann and Fimm's Attention battery (TEA) (P<0.01) and reverse digit span (DS, P<0.05). Factor F2 was positively related to age at injury, time since injury (TSI) (P<0.05) and CESD and STAI scores (P<0.001). Factor F3 was related to DA (P<0.01) TL scores and TSI (P<0.05). Factor F4 was related to TL, TMT, DA, flexibility, DS (P<0.05), TSI, duration of post-traumatic amnesia, CES-D score (P<0.05) and STAI scores (P<0.01). Factor F5 was related to GCS, DA (P<0.05), and reaction time on the subscales IN and Go/nogo (GO) of the TEA battery (P<0.01). The NRS-R total score was related to CESD, STAI scores, TMT score, DA (P<0.01) and TL score, IN and GO scores and TSI (P<0.05). DISCUSSION: As McCauley et al found with the English version of the NRS-R, significant relationships were found between NRS-R factor scores, cognitive tests and emotional scales. Relationships were also found between factor scores and indicators of injury severity and time since injury. These data suggest fair concurrent validity of the NRS-R. PMID- 15748771 TI - [Botulinum toxin and medical liability: is the patient sufficiently informed?]. AB - INTRODUCTION: The use of botulinum toxin injection therapy is soaring significantly today, with an ever-wider field of applications despite well-known side effects of the treatment. This article aims at analysing the medicolegal practices of practitioners who use this therapy, especially the information given to patients and finding a common practice for providing that information. METHODS: We sent a questionnaire to 340 practitioners who might use the therapy (physiatrists, neurologists, ophthalmologists, ENT specialists, plastic surgeons) working in hospitals and in physical therapy and rehabilitation centres in France. Besides mentioning the possible side effects of the therapy, the questionnaire focused on how such information was transmitted before the injection. RESULTS: Data collection and analysis were performed by use of a spreadsheet software programme. A total of 124 questionnaires were analysed. We did not analyse the items dealing with side effects. Sixty-five percent of the responders said they did not seek statutory authorisation for injections. Only 31% provided written, detailed information and 12% required a signed consent form. Complaints were rare, approximately 12%, were written or verbal, and were always dismissed. DISCUSSION: Side effects after botulinum toxin injection are clearly described in the medical literature. Therefore, it is of utmost importance for this product to be used therapeutically and only by experienced therapists who will carefully respect the product's standard rules of use and inform their patients to the best of their ability. Issuing a detailed letter of information describing all the side effects seems necessary. We suggest a model information letter such as that provided to the patients in our facility. CONCLUSION: Botulinum toxin is a very worthwhile product for numerous abnormalities but has side effects, often brief, at the site of the injection. Therefore it is our duty to inform patients effectively. PMID- 15748772 TI - [Injection of botulinum toxin and medical liability. Is the patient sufficiently informed?]. PMID- 15748773 TI - [Isokinetic strength measurement and training of the shoulder: methodology and results]. AB - OBJECTIVE: To evaluate the contribution of isokinetic methods of shoulder strength measurement and training. METHOD: A Medline search of English and French publications, including referenced articles, allowed us to analyse non-indexed publications. Eighty-seven articles were retained for analysis. RESULTS: The isokinetic evaluation of the shoulder is valid. Although reproducibility of shoulder evaluation is inferior to that of the knee, it is nevertheless satisfactory when a rigorous test method is used. Normal values for the rotators, abductors-adductors, and extensors-flexors depend on diverse parameters such as age, gender, fat mass, and the type and intensity of physical activity. The agonist to antagonist ratio is particularly informative in pathological conditions. The ratio is modified in cases of impingement syndrome and shoulder instability, and this modification appears to be a cause rather than a consequence of pathologic features. The ratio generally remains modified post surgery, and normalization must be a major focus of post-surgery rehabilitation. CONCLUSION: Isokinetic measurement, particularly disturbances in the agonist antagonist balance, is a reference method for evaluating shoulder muscle strength and detecting deficits in specific muscle groups seen in certain shoulder abnormalities. Such measurement is a valuable tool for orienting rehabilitation towards the deficient muscle groups, complements classical techniques of muscle strengthening, and is an accurate means for following the rehabilitation progress. PMID- 15748774 TI - [Detraining and retraining after injury in a high-level cyclist]. AB - OBJECTIVE: To define retraining after injury in a high-level cyclist by taking into account the consequences of detraining. METHOD: From three clinical cases and from the analysis of the consequences of detraining, three principles of retraining were determined. RESULTS: 1. The high-level cyclist is not protected and loses cycling capacity after four weeks of inactivity. The delay in recovery is longer the higher the adaptations. 2. Recovery of cycling capacity is based on bicycle exercises that are greater in intensity than quantity, taking into account delays in injury consolidation. 3. Retraining requires appreciating the individual physiological level by evaluating force and endurance before envisaging the resumption of training and competition. CONCLUSION: The injury of a high-level cyclist is at the origin of detraining, which has been evaluated so that sports rehabilitation may enable the cyclist to find a previous state without relapse, complication or overtraining. PMID- 15748775 TI - [Constituted adult equine from the ankle: surgical treatment by Ilizarov external fixator]. AB - INTRODUCTION: Ilizarov's external fixator indications are classically used in children to lengthen the lower limbs. This technique is used for adults to gradually correct articular limitations, especially in the knee, ankle and foot. METHOD AND RESULTS: We report a case of a 53-year-old patient with distal microangiopathy of the lower limbs leading to amputation of the toes and a bilateral, direct, irreducible, isolated equine feature? from the back foot to 0/50/60 degrees? on retraction of the Achilles tendon. The aim of patient management was to regain walking ability. Because of vascular and cutaneous fragility, the classical surgical treatment was contra-indicated. Progressive reduction by Ilizarov's external fixator was tried. Dorsal inflection was obtained by twice-daily screwing until a radiological angle of 90 degrees was obtained between the tibia and talus. At ten weeks post-surgery, the dorsal voluntary inflection ankle amplitude was symmetrical to 0/10/60 degrees. The patient walked with orthopaedic shoes compensating for the few degrees of residual equine. DISCUSSION: Correction of adult equine without osseous deformation by Ilizarov's external fixator should be considered when conventional surgery is contra-indicated. PMID- 15748776 TI - Vitamin E levels, cognitive impairment and dementia in older persons: the InCHIANTI study. AB - There is conflicting evidence that antioxidants contribute to maintaining cognitive function in elderly subjects. We investigated whether vitamin E plasma levels are related to the presence of dementia and cognitive impairment in a population-based cohort study conducted in Italy. A total of 1033 participants aged at least 65 years received clinical and neuropsychological examinations, donated blood for vitamin E analysis and had their diets assessed. Participants with plasma vitamin E levels in the bottom tertile had a significantly higher probability of being demented (OR 2.6, 95% CI 1.0-7.1) and also of suffering from cognitive impairment (OR 2.2, 95% CI 1.2-4.2) compared to those in the highest vitamin E tertile after adjustment for age, gender, education, lipid levels, energy intake, vitamin E intake, and smoking. This study supports the notion that higher vitamin E plasma levels might provide significant protection against cognitive impairment and dementia in elderly subjects. PMID- 15748777 TI - Caloric restriction attenuates Abeta-deposition in Alzheimer transgenic models. AB - Dietary influences on Alzheimer disease (AD) are gaining recognition. Because many aging processes are attenuated in laboratory mammals by caloric restriction (CR), we examined the effects of short-term CR in two AD-transgenic mice, APP(swe/ind) (J20) and APP(swe) + PS1(M146L) (APP + PS1). CR substantially decreased the accumulation of Abeta-plaques in both lines: by 40% in APP(swe/ind) (CR, 6 weeks), and by 55% in APP + PS1 (CR, 14 weeks). CR also decreased astrocytic activation (GFAP immunoreactivity). These influences of CR on AD transgenic mice are consistent with epidemiological reports that show that high caloric diets associate with the risk of AD, and suggest that dietary interventions in adult life might slow disease progression. PMID- 15748778 TI - Interaction of herpesviridae, APOE gene, and education in cognitive impairment. AB - While high age, low level of education and APOE epsilon4 allele are known to predict dementia, there is recent data suggesting that certain viruses and subtypes of APOE epsilon3 could be involved, too. We investigated these relationships in a home-dwelling cohort of 357 elderly people with various cardiovascular diseases (DEBATE study). MMSE score below 24 was used to define cognitive impairment (n = 58). When adjusted for age and the presence of diabetes, multivariate analysis demonstrated maximally increased risk of cognitive impairment in association with a combination of three factors: seropositivity for herpesviridae, presence of APOE epsilon4, and low education (risk ratio 6.1, 95% CI 2.4-15.2). In the subcohort of APOE3/3 individuals (n = 216) homozygosity for the -219G epsilon3 haplotype showed a similar association (risk ratio 8.8, 95% CI 2.6-29.8). These results demonstrate an interaction of specific genetic (APOE) and environmental (education and herpesviridae) risk factors in the development of cognitive impairment and indicate that not only the epsilon4 allele of APOE but also the epsilon3 haplotype is a risk factor for dementia. PMID- 15748779 TI - The association between promoter polymorphism of the interleukin-10 gene and Alzheimer's disease. AB - The importance of the role of inflammation has been suggested in the pathogenesis of Alzheimer's disease (AD). Interleukin-10 (IL-10) is an anti-inflammatory cytokine that may modulate the progression of the disease through the inhibition of the action of pro-inflammatory cytokines. In this study, three polymorphisms in the regulatory region of the IL-10 gene (-1082, -819 and -592) in 95 Chinese AD patients and 117 age-matched healthy Chinese subjects were investigated. We found that among the Chinese population, the A and C alleles at the -592 position are strongly linked to the T and C alleles at the -819 position, respectively. A strong association with AD was found for these two IL-10 polymorphisms, which are in complete linkage disequilibrium (-592C and -819C), and the odds ratio of AD is 4.03 (95% CI 1.23-13.23; p = 0.011). The functional significance of the IL-10 genotype was further supported by the significant association between plasma IL 10 concentrations and genotypes that were found in an independent sample of 160 healthy male volunteers. No interaction effect between the ApoE and IL-10 genotypes is found. Therefore, we concluded that the functional polymorphisms of the IL-10 gene act as a risk factor for AD. PMID- 15748780 TI - Genetic association between matrix metalloproteinase MMP-9 and MMP-3 polymorphisms and Japanese sporadic Alzheimer's disease. AB - Recent studies suggested that matrix metalloproteinases (MMPs) might play an important role in the pathophysiology of Alzheimer's disease (AD). MMP-9 and MMP 3 are reported to degrade amyloid beta and have several functional polymorphisms associated with other common diseases. Four common polymorphisms in each of MMP-9 and MMP-3 were examined in AD cases and normal control individuals. Common polymorphisms of MMP-9, rs3918248, rs2664538, rs2250889 and rs2274756 showed no association with risk for AD. We observed strong linkage disequilibrium (LD) between rs2664538 and rs2250889 in our Japanese samples. The polymorphisms of MMP 3; 5A/6A insertion polymorphism in the promoter, rs3025079, rs520540 and rs679620 also did not influence risk for AD. LD of the 5A/6A polymorphism with rs679620 was relatively strong. These results suggest that the common polymorphisms of MMP 9 and MMP-3 investigated here are not associated with AD. PMID- 15748781 TI - Tau truncation during neurofibrillary tangle evolution in Alzheimer's disease. AB - The microtubule-associated protein, tau, is a highly soluble molecule that is nonetheless capable of self-association into filamentous deposits characteristic of a number of neurodegenerative diseases. This state change is thought to be driven by phosphorylation and/or C-terminal truncation events resulting in intracellular inclusions, such as the neurofibrillary tangles (NFTs) in Alzheimer's disease (AD). Previously, we reported the existence of a novel truncation event, cleavage at aspartic acid(421), presumably by a caspase, and also described a monoclonal antibody (Tau-C3) specific for tau cleaved at this site. Here, we report the timing of this cleavage event relative to other antibody-targeted alterations in the tau molecule during the course of NFT evolution in AD. Immunohistochemical studies indicate that cleavage at aspartic acid(421) occurs after formation of the Alz50 epitope but prior to formation of the Tau-66 epitope and truncation at glutamic acid(391) (formation of the MN423 epitope). Thus, creation of the Tau-C3 epitope appears to occur relatively early in the disease state, contemporaneous with the initial Alz50 folding event that heralds the appearance of filamentous tau in NFTs, neuropil threads, and the dystrophic neurites surrounding amyloid plaques. PMID- 15748782 TI - Human-like rodent amyloid-beta-peptide determines Alzheimer pathology in aged wild-type Octodon degu. AB - It is generally accepted that human Alzheimer's disease (AD) neuropathology markers are completely absent in rodent brains. We report here that an aged wild type South American rodent, Octodon degu, expresses neuronal beta-amyloid precursor protein (beta-APP695) displaying both intracellular and extracellular deposits of amyloid-beta-peptide (Abeta), intracellular accumulations of tau protein and ubiquitin, a strong astrocytic response and acetylcholinesterase (AChE)-rich pyramidal neurons. The high amino acid homology (97.5%) between deguAbeta and humanAbeta sequences is probably a major factor in the appearance of AD markers in this aged rodent. Our results indicate that aged O. degu constitutes the first wild-type rodent model for neurodegenerative processes associated to AD. PMID- 15748783 TI - Estradiol or estradiol/progesterone treatment in older women: no strong effects on cognition. AB - The relevance of estrogens for cognition in older women is still debated. In this double-blind experiment hysterectomized women (age 58-75 years) received placebo (n = 13), estradiol (n = 12) or estradiol/progesterone (n = 10) treatment. Cognitive testing (nine different tests) took place at baseline, after 4 and 24 weeks of treatment. Strong hormone increases occurred in both active treatment groups. However, no beneficial effects in any of the cognitive tests could be detected. This study, therefore, does not support the notion that treatment with sex hormones has beneficial effects on cognition in older hysterectomized women. The human brain might loose its responsiveness to gonadal steroids with aging or prolonged hormone depletion. PMID- 15748784 TI - Regulation of cytochrome oxidase activity in the rat forebrain throughout adulthood. AB - Measures of metabolic activity can provide useful indices of the effects of aging on neural function, since sustained changes in neural activity alter metabolic demand and the activity of metabolic enzymes. Previous reports of effects of aging on key enzymes for oxidative metabolism are mixed, however, with some reports that activity declines in the aging brain and others that activity remains stable or increases. We used high-resolution, quantitative histochemistry to test whether cytochrome oxidase (CO) activity changes in the forebrain during adulthood and senescence, measuring activity in each layer of the hippocampus and several cerebral cortical areas. In most forebrain regions, average cytochrome oxidase activity was slightly higher in middle-aged than in young adult rats but did not differ between middle-aged and old rats. Thus, there was no significant change in cytochrome oxidase activity with senescence. Additional analyses indicated that cytochrome oxidase activity is regulated regionally in the brain, as well as focally, and that differences in regional regulation may contribute to variation in CO activity among individuals, which was greater in young and old rats than in middle-aged animals. PMID- 15748785 TI - Age-related metabolic changes in the upper brainstem tegmentum by MR spectroscopy. AB - Several neurodegenerative disorders have a profound metabolic and structural impact on the brainstem. MR spectroscopy provides metabolic information non invasively and has the potential to characterize the changes associated with normal aging and differentiate them from neurodegenerative alterations. The present work was aimed at studying the upper brainstem tegmentum at the midbrain and pontine levels in 57 adult normal volunteers, aged 23-79 years, with long echo time proton MR spectroscopy to evaluate possible regional differences and the effect of age. Higher ratios of N-acetyl aspartate (NAA)/total creatine (Cr) and choline-containing compounds (Cho)/Cr were observed in the pons compared to the midbrain, resulting from higher net NAA and Cho content. In the midbrain, there was a linear decline of NAA and Cho with age in subjects over 50, most probably related to neuronal tissue loss. In the pons, such an aging effect was not observed, with subjects over 50 showing higher Cr and Cho than the under-50 subjects. Our findings provided evidence of regional differences and suggest different effects of age on the two studied brainstem segments, hitherto undescribed. PMID- 15748786 TI - Aging and subcellular localization of m2 muscarinic autoreceptor in basalocortical neurons in vivo. AB - By using immunohistochemical approaches at the light and electron microscopic levels, we have shown that aging modifies the subcellular distribution of the m2 muscarinic autoreceptor (m2R) differentially at somato-dendritic postsynaptic sites and at axonal presynaptic sites in cholinergic basalocortical neurons, in vivo. In cholinergic perikarya and dendrites of the nucleus basalis magnocellularis (NBM), aging is associated with a decrease of the density of m2R at the plasma membrane and in the cytoplasm, suggesting a decrease of the total number of m2R in the somato-dendritic field. In contrast, the number of substance P receptors per somato-dendritic surface was not affected. In the frontal cortex (FC), we have shown a decrease of cytoplasmic m2R density also leading to a decrease of the number of m2R per surface of varicosities but with no change of the density of m2R at the membrane. Our results suggest that the decrease of m2R in the somato-dendritic field of the NBM, but not a modification of the number of presynaptic m2 autoreceptors at the plasma membrane in the FC, could contribute to the decrease of the efficacy of cholinergic transmission observed with aging in the rat. PMID- 15748787 TI - Aging and cholinergic deafferentation alter GluR1 expression in rat frontal cortex. AB - Previously, we demonstrated that plasticity of frontal cortex is altered in aging rats: lesions of the nucleus basalis magnocellularis (NBM) produce larger declines in dendritic morphology in frontal cortex of aged rats compared to young adults. Cholinergic afferents from the NBM modulate glutamatergic transmission in neocortex, and glutamate is known to be involved in dendritic plasticity. To begin to identify possible mechanisms underlying age-related differences in plasticity after NBM lesion, we assessed the effect of cholinergic deafferentation on expression of the AMPA receptor subunit GluR1 in frontal cortex of young adult and aging rats. Young adult, middle-aged, and aged rats received sham or 192 IgG-saporin lesions of the NBM, and an unbiased stereological technique was used to estimate the total number of intensely GluR1 immunopositive neurons in layer II-III of frontal cortex. While the number of GluR1-positive neurons was increased in both middle-aged and aged rats, lesions markedly increased the number of intensely GluR1-immunopositive neurons in frontal cortex of young adult rats only. This age-related difference in lesion induced expression of AMPA receptor subunit protein could underlie the age related differences in dendritic plasticity after NBM lesions. PMID- 15748788 TI - Age-dependent changes of gene expression in the Drosophila head. AB - Previous gene expression profiling studies in Drosophila have provided clues for understanding the aging process at the gene expression level. For a detailed understanding, studies of specific regions of the body are necessary. We therefore employed microarray analysis to examine gene expression changes in the Drosophila head during aging. Six hundred and eighty-four of the 5405 genes present in the microarray showed significant age-dependent changes as determined by significance analysis of microarray (SAM) (q < 0.05). The biological significance of the changes was analyzed using the gene annotations provided by the Gene Ontology Consortium. Major changes involved genes affecting energy metabolism (proton transport, energy pathways, oxidative phosphorylation) and neuronal function, especially responses to light. Genes involved in protein catabolism and several other metabolic processes also showed age-dependent changes. Most of the changes were reductions in gene expression and occurred before day 13 of adult life. After day 13, the age-dependent gene expression changes were relatively smaller than earlier life. Interestingly, the two biological processes of major gene expression changes are related to the two known environmental changes that increase life span in Drosophila: caloric restriction and light reduction. Our findings suggest that light signaling and energy metabolism may be important biological processes affected by aging and be interesting targets for the further investigation related to the longevity in Drosophila. PMID- 15748789 TI - Preservation of hippocampal volume throughout adulthood in healthy men and women. AB - To address controversies regarding the effect of age on the hippocampus, volumes of hippocampus and a comparison structure, temporal cortex, were measured on magnetic resonance imaging (MRI) in 84 healthy men and 44 healthy women (20-85 years). Neither men nor women showed significant correlations between hippocampal volumes and age, despite significant age-related decline in temporal volumes. Absence of hippocampus age relationships endured when restricting analyses to older individuals (> or =50 years) and considering menopause and hormone replacement therapy. PMID- 15748791 TI - Spontaneous fos expression in the suprachiasmatic nucleus of young and old mice. AB - The senescence-accelerated mouse (SAMP8) is an animal model of aging that displays an array of circadian rhythm disruptions as early as 7 months of age. The present study explored the physiological basis for age-related changes in circadian rhythms by measuring c-Fos immunostaining. Cellular activity in the SCN "core" and "shell" was examined for 2-, 7-, and 12-month-old SAMP8 at circadian times (CTs) 2 and 14. Consistent with previous studies in rats, we observed higher levels of cellular activity at CT2 than at CT14, and higher levels of activity in the "shell" than in the "core" of the SCN. However, there was no effect of age on the pattern of cellular activity in either the "core" or the "shell" of the SCN. These results are discussed in the context of current research on spontaneous and light-induced c-Fos expression in the SCN of rodents. PMID- 15748790 TI - Place cells of aged rats in two visually identical compartments. AB - Aged rats perform poorly on spatial learning tasks, a cognitive impairment which has been linked to the failure of hippocampal networks to fully encode changes in the external environment [Barnes CA, Suster MS, Shen J, McNaughton BL. Multistability of cognitive maps in the hippocampus of old rats. Nature 1997;388(6639):272-5; Wilson IA, Ikonen S, Gureviciene I, McMahan RW, Gallagher M, Eichenbaum H, et al. Cognitive aging and the hippocampus: how old rats represent new environments. J Neurosci 2004;24(15):3870-8]. To examine whether the impairment in hippocampal processing extends to conditions in which self motion provides the cues for environmental change, we have analyzed spatial firing patterns of hippocampal pyramidal neurons in young and aged rats, as well as in young rats with selective cholinergic lesions, another model of cognitive aging. The rats walked between two visually identical environments, pitting self motion cues that indicated environmental change against visual inputs that indicated no differences between environments. Our results indicated that place cells in both aged and cholinergic-lesioned rats were equally likely as those of young rats to create new spatial representations in the second compartment. These findings suggest that the hippocampal network of aged rats is able to process changes in internally generated cues without rigidity, but that incomplete processing of external landmark cues may lead to impaired spatial learning. PMID- 15748793 TI - Skin irritation: prevalence, variability, and regulatory classification of existing in vivo data from industrial chemicals. AB - In vivo rabbit data for skin irritation registered in the European New Chemicals Database (NCD) and an ECETOC Database were evaluated to characterise the distribution of irritation potential among chemicals and to assess the variability of the animal test. These databases could be used to determine experimental and rudimentarily within-laboratory variability, but not between laboratory variability. Our evaluation suggests that experimental variability is small. Using two classification systems--the system currently used in Europe and the Globally Harmonised System (GHS)--the prevalence of skin irritation data obtained from NCD was analysed. This analysis revealed that out of 3121 chemicals tested, less than 10% showed an irritation potential in rabbits which would require an appropriate hazard label and 64% did not cause any irritation. Furthermore, it appears that in practical use the European classification system introduces bias towards overclassification. Based on these findings, we conclude, that the classification systems should be refined taking prevalence into account. Additionally, prevalence should be incorporated into the design and analysis of validation studies for in vitro test methods and in the definition of testing strategies. PMID- 15748792 TI - Age-related decline in caloric intake and motivation for food in rhesus monkeys. AB - Human studies have documented age-related declines in caloric intake that are pronounced at advanced ages. We examined caloric intake from a longitudinal study of aging in 60 male and 60 female rhesus monkeys (Macaca mulatta) collected for up to 10 years. Monkeys were provided a standardized, nutritionally fortified diet during two daily meals, and intake was measured quarterly. About half of the monkeys were on a regimen of caloric restriction (CR) representing about a 30% reduction in caloric intake compared to controls (CON) of comparable age and body weight. CR was applied to determine if this nutritional intervention retards the rate of aging in monkeys similar to observations in other mammalian studies. Following reproductive maturity at 6 years of age, there was a consistent age related decline in caloric intake in these monkeys. Although males had higher intake than females, and CON had higher intake compared to CR, the sex and diet differences converged at older ages (>20 years); thus, older CR monkeys were no longer consuming 30% less than the CON. When adjusted for body weight, an age related decline in caloric intake was still evident; however, females had higher intake compared to males while CR monkeys still consumed less food, and again differences converged at older ages. Motivation for food was assessed in 65 of the monkeys following at least 8 years in their respective diet groups. Using an apparatus attached to the home cage, following an overnight fast, monkeys were trained to reach out of their cage to retrieve a biscuit of their diet by pushing open a clear plastic door on the apparatus. The door was then locked, and thus the biscuit was irretrievable. The time spent trying to retrieve the biscuit was recorded as a measure of motivation for food. We observed an age-related decline in this measure, but found no consistent differences in retrieval time between CR and CON groups of comparable age and time on diet. The results demonstrate an age related decline in food intake and motivation for food in rhesus monkeys paralleling findings in humans; however, we found no evidence that monkeys on a long-term CR regimen were more motivated for food compared to CON. Examining the relationship of selected blood proteins to food intake following 7-11 years on the study, we found a negative correlation between globulin and intake among males and females after accounting for differences in age. In addition, a positive correlation was observed between leptin and intake in males. PMID- 15748795 TI - Quantitative measurement of protein digestion in simulated gastric fluid. AB - The digestibility of novel proteins in simulated gastric fluid is considered to be an indicator of reduced risk of allergenic potential in food, and estimates of digestibility for transgenic proteins expressed in crops are required for making a human-health risk assessment by regulatory authorities. The estimation of first order rate constants for digestion under conditions of low substrate concentration was explored for two protein substrates (azocoll and DQ-ovalbumin). Data conformed to first-order kinetics, and half-lives were relatively insensitive to significant variations in both substrate and pepsin concentration when high purity pepsin preparations were used. Estimation of digestion efficiency using densitometric measurements of relative protein concentration based on SDS-PAGE corroborated digestion estimates based on measurements of dye or fluorescence release from the labeled substrates. The suitability of first order rate constants for estimating the efficiency of the pepsin digestion of novel proteins is discussed. Results further support a kinetic approach as appropriate for comparing the digestibility of proteins in simulated gastric fluid. PMID- 15748794 TI - Reassessment of ecotoxicities of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane in Japanese quail under the OECD draft new avian one-generation reproduction test guideline. AB - Ecotoxicological hazards of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (p,p' DDT) were investigated by a one-generation reproduction study using Japanese quail (Coturnix coturnix japonica) under an Organization for Economic Co operation and Development (OECD) draft new test guideline 206 following acute and subchronic toxicity studies. In the subchronic feeding toxicity study, tremors, convulsions, and deaths were observed with a clear sex difference, males being more susceptible than females. The estimated total number of sperm tended to decrease in a dose-dependent manner at the end of 6-week treatment. In the one generation reproduction study conducted at dose levels of 0, 6, 30, and 150 ppm, the estimated total number of sperm tended to decrease in a dose-dependent manner with a significant difference at 150 ppm. Tremors were observed in the majority of hatchlings in the 150 ppm group and at lower incidences in the 30 ppm group. Significantly high mortality rate in chicks persisted from treatment week 3-6 in the 150 ppm group and at treatment weeks 4 and 5 in the 30 ppm group. Despite of these severe adverse effects of p,p'-DDT on hatchlings and chicks, fertilization, egg laying, eggshell thickness or embryonic development was hardly impaired by p,p'-DDT or its metabolites. From these results, it appears that the OECD draft new avian one-generation reproduction test guideline is effective for ecological hazard assessment of chemicals. PMID- 15748796 TI - Smoke composition and predicting relationships for international commercial cigarettes smoked with three machine-smoking conditions. AB - The study objectives were to determine the effects of smoking machine puffing parameters on mainstream smoke composition and to express those effects as predicting relationships. Forty-eight commercial Philip Morris USA and Philip Morris International cigarettes from international markets and the 1R4F reference cigarette were machine-smoked using smoking conditions defined by the International Organization of Standardization (ISO), the Massachusetts Department of Public Health (MDPH), and Health Canada (HC). Cigarette tobacco fillers were analyzed for nitrate, nicotine, tobacco-specific nitrosamines (TSNA), and ammonia. Mainstream yields for tar and 44 individual smoke constituents and "smoke pH" were determined. Cigarette constituent yields typically increased in the order ISO5000 mg/kg (oral), >2000 mg/kg (dermal), and >26,411 ppm (inhalation: reversible anaesthetic effects, e.g., lethargy, seen at this exposure concentration). Other short-term tests (skin and eye irritation, skin sensitisation, genotoxicity tests in vitro and in vivo, cardiac sensitisation) were also performed, and no hazardous properties were identified. Effects of repeated exposure by inhalation were examined in rats over test periods of 5, 14, 28, and 90 days. Effects on embryo foetal development in the rat have also been studied. The 28-day, 90-day and developmental studies were performed using nominal HFPE concentrations of 1000, 3300, and 10,000 ppm (6h/day: actual exposures confirmed by test atmosphere analysis), and the highest tested concentration proved to be an NOAEL in each study. Major observed effects were elevated urinary (inorganic) fluoride levels and increased liver weights with centrilobular hepatocyte hypertrophy (considered an adaptive response, linked to hepatic metabolism of absorbed material). PMID- 15748798 TI - Analyses of neurobehavioral screening data: dose-time-response modeling of continuous outcomes. AB - Neurotoxic effects are a non-cancer endpoint for health risk, and neurobehavioral screening tests can serve as a first tier investigation of neurotoxicity [US EPA, Federal Register 63 (1998) 26926]. Analysis of neurobehavioral screening data such as those of the functional observational battery (FOB) traditionally relies on analysis of variance (ANOVA). ANOVA is designed to detect whether there are dose-effects, but does not model the underlying dose-response relationship and subsequent risk assessment fails to utilize the shape of the underlying dose response. In contrast, dose-response modeling interpolates toxic effects between experimental points, and permits prediction of toxic effects within the experimental range. Additionally it is also a prerequisite for estimating a benchmark dose. This paper discusses dose-time-response modeling of longitudinal neurotoxicity data and illustrates the methods using three continuous FOB outcomes from an EPA study involving acute exposure to triethyltin (TET). Several mathematical functions are presented as candidate dose-time-response models. The use of random effects is discussed to characterize inter-subject variation. The results indicate that it is feasible to use simple mathematical functions to model empirical dose-time-response observed in existing longitudinal neurotoxicological data. Further research is needed on the types of design and data required to reliably approximate the true underlying dose-time-response. PMID- 15748799 TI - Fire assisted pastoralism vs. sustainable forestry--the implications of missing markets for carbon in determining optimal land use in the wet-dry tropics of Australia. AB - Using Cape York Peninsula, Queensland, Australia as a case study, this paper combines field sampling of woody vegetation with cost-benefit analysis to compare the social optimality of fire-assisted pastoralism with sustainable forestry. Carbon sequestration is estimated to be significantly higher in the absence of fire. Integration of carbon sequestration benefits for mitigating future costs of climate change into cost-benefit analysis demonstrates that sustainable forestry is a more socially optimal land use than fire-assisted pastoralism. Missing markets for carbon, however, imply that fire-assisted pastoralism will continue to be pursued in the absence of policy intervention. Creation of markets for carbon represents a policy solution that has the potential to drive land use away from fire-assisted pastoralism towards sustainable forestry and environmental conservation. PMID- 15748800 TI - Capacity building for sustainability: towards community development in coastal Scotland. AB - Capacity building is widely recognised as a central feature of Integrated Coastal Zone Management (ICZM). Yet, although increased community capacity can potentially empower local communities with the ability to tackle the impacts of socio-economic change, an evaluation of ICZM performance reveals a limited interest in advancing community development initiatives. This weakness is particularly apparent in Scotland. Whilst coastal communities in Scotland have been found to suffer from a range of significant socio-economic problems, Scottish approaches to ICZM have failed to improve local development trajectories. In response, this paper considers the degree to which community based partnerships can assist in the delivery of sustainable development benefits. PMID- 15748801 TI - Comparisons of pastoralists perceptions about rangeland resource utilisation in the Middle Awash Valley of Ethiopia. AB - Pastoralism is the most dominant land use form in the arid rangelands of Sub Saharan Africa, but this rangeland-based lifestyle is under threat. As a consequence a study was conducted in the Middle Awash Valley of Ethiopia with the main objectives of assessing and comparing the broad perceptions of two pastoral groups (the Oromo ethnic group living in Kereyu-Fantale and the Afar ethnic group living in Awash-Fantale) on the usage of the existing rangeland resources, and their views on constraints and possible solutions. Data were collected from 90 Oromo and 55 Afar households. Despite the difference in ethnicity both of these groups share common problems. They derive their main income from the sale of animals and animal products, but with the difference that milk products rank first in the case of the Afar and last in the case of the Oromo. Both pastoral groups depend heavily on native grasses for animal feed and to a lesser extent on woody plants as a source of browse. The majority of respondents were of the opinion that the condition of the rangelands is poor, mainly due to overgrazing, droughts and increases in the human population. Availability of water is also regarded as a problem, mainly by the Oromo. Migration is the first measure taken to solve shortages of livestock feed, but many of the pastoralists replied that migration is an undesirable practise which is mostly done out of necessity. Because of the limited resources most respondents of both groups still prefer communal land tenure where resources are shared. It is concluded that the problems facing the pastoralists in the Middle Awash Valley have been created over many years and the solutions will also require time. With the current approach of the communal grazing systems, sustainable utilisation of the rangeland ecosystem is not possible. Solutions to the poor condition of the existing rangelands will require a definite commitment and full participation not only of the pastoralists, but also of government and non-governmental organisations that are directly or indirectly involved in rangeland resources utilisation, management, conservation and other related activities. PMID- 15748802 TI - Delivering sustainability therapy in sustainable development projects. AB - This paper explores the apparent contradiction between the 'linearity' of most Sustainable Development projects, with time-bound and defined outputs achieved at a fixed cost, and an implied 'circularity' of the theory whereby there is no 'end'. Projects usually have clear parameters within which they are implemented, and the inclusion of elements such as the need for accountability, measurable impact and 'value for money' have grown in importance. It could be argued that we live in a 'projectified' and therefore linear world. The paper explores the potential contradiction between 'linearity' and 'circularity', and suggests that one way around this is to frame the project within a form of the Kolb Learning Cycle heuristic. This will facilitate a rationalisation from those implementing the sustainable development project as to why decisions are being made and for whom. If these questions are opened up to the project stakeholders, including beneficiaries, then the Kolb cycle could encourage learning and understanding by all involved. It could also provide Sustainability Therapy to those trapped in processes, which they find orthogonal to their own perceptions. It is suggested that such learning, therapy and reflective practice should be a valid output of the sustainable development project, although typically the focus is only upon the final outputs and how they feed into policy. Ironically funders would be well advised to take a broader perspective in order to achieve true 'value for money' within such projects, even if learning is not an easily measurable or tangible outcome. These points are explored within the context of the wider literature and experience with a sustainable development project undertaken in Malta. PMID- 15748803 TI - Selection of hydrothermal pre-treatment conditions of waste sludge destruction using multicriteria decision-making. AB - The effectiveness of hydrothermal treatment for the destruction of the organic content of sludge waste was investigated. The sludge sampled in this study contained approximately 2% solids. The experimental program consisted of hydrothermal treatment experiments conducted in a batch reactor at temperatures between 100 and 250 degrees C, with the addition of an oxidant (hydrogen peroxide) in the range of 0-150% with reference to TCOD, and reaction times of up to 60 min. The results suggested that the availability of oxidant, reaction temperature and reaction time were the determining factors for COD removal. A significant fraction of the COD remaining after treatment consisted of the dissolved COD. The results confirmed that hydrothermal treatment proceeds through hydrolysis resulting in the production of dissolved organic products followed by COD removal through oxidation. Two MCDM chemometrics methods, PROMETHEE and GAIA, were applied to process the large data matrix so as to facilitate the selection of the most suitable hydrothermal conditions for sludge destruction. Two possible scenarios were produced from this analysis-one depended on the use of high temperatures and no oxidant, while the second offered a choice of compromise solutions at lower temperatures but with the use of at least some oxidant. Thus, for the final choice of operating conditions, the decision maker needs local knowledge of the costs and available infrastructure. In principle, such information could be added as further criteria to the data matrix and new rankings obtained. PMID- 15748804 TI - Co-management: concepts and methodological implications. AB - Co-management, or the joint management of the commons, is often formulated in terms of some arrangement of power sharing between the State and a community of resource users. In reality, there often are multiple local interests and multiple government agencies at play, and co-management can hardly be understood as the interaction of a unitary State and a homogeneous community. An approach focusing on the legal aspects of co-management, and emphasizing the formal structure of arrangements (how governance is configured) runs the risk of neglecting the functional side of co-management. An alternative approach is to start from the assumption that co-management is a continuous problem-solving process, rather than a fixed state, involving extensive deliberation, negotiation and joint learning within problem-solving networks. This presumption implies that co management research should preferably focus on how different management tasks are organized and distributed concentrating on the function, rather than the structure, of the system. Such an approach has the effect of highlighting that power sharing is the result, and not the starting point, of the process. This kind of research approach might employ the steps of (1) defining the social ecological system under focus; (2) mapping the essential management tasks and problems to be solved; (3) clarifying the participants in the problem-solving processes; (4) analyzing linkages in the system, in particular across levels of organization and across geographical space; (5) evaluating capacity-building needs for enhancing the skills and capabilities of people and institutions at various levels; and (6) prescribing ways to improve policy making and problem solving. PMID- 15748805 TI - Sorption studies of mixed chromium and chlorinated ethenes at the field and laboratory scales. AB - The sorption and desorption behavior of hexavalent chromium and chlorinated ethenes in a sandy ground water aquifer with a low reduction capacity was evaluated by performing a variety of analyses and experiments at the laboratory (batch and column studies) and field (in situ injection-withdrawal experiment) scales. The partitioning coefficients determined from the field and laboratory experiments are generally similar. Both sets of experiments yielded relatively low partition coefficients for chromium and chlorinated ethenes. The column studies and injection-withdrawal experiment indicate that chromium has the potential to leach from aquifer sediments and act as a secondary source of contamination. However, the magnitude of the secondary source effect is not significant due to low concentrations of leachable contamination. The chromium sorption isotherm data were also simulated using the triple layer surface complexation model (TLM). The isotherm data were modeled using the TLM, illustrating the applicability of geochemical modeling for sorption of chromium to these sediments under variable pE-pH conditions. PMID- 15748806 TI - Species diversity or biodiversity? AB - Species diversity and biodiversity are widely used terms in ecology and natural resource management. Despite this, they are not easily defined and different authors apply these terms with varying connotations. The term biodiversity, in particular, has the dubious honour of being widely used but rarely defined. Is it simply the number of species or is it something more? Here I consider what these terms might really mean and their value. I also briefly discuss the rationale for studying and protecting species diversity or biodiversity. PMID- 15748807 TI - Selection of immunodominant fragments from envelope gene for vaccine against Japanese encephalitis virus in DNA priming-protein boosting protocols. AB - Fragmentation of E gene of JEV into smaller fragments, none of the fragments either in plasmids form or in recombinant protein form can induce optimal protection against the virus infection. It is only when DNA priming-protein boosting strategies are used then the N-terminal E(A) and the C-terminal E(B) showed full protection against JEV as those induced by commercial vaccine, provided both fragments are preceded in the N-terminal by a signal peptide M(15) derived from C-terminal of prM gene in JEV genome. When the subfragments of E(A): E(A1) and E(A2) and E(B): E(B1) and E(B2) are tested, only E(A1) subfragment can replace E(A) in protein boosting to induce optimal protection against JEV, E(A2), E(B1), E(B2) in plasmid or protein forms are not. Therefore, along the E gene (978-2330 bp) N-terminal, E(A1) (978-1580 bp) and C-terminal E(B) (1851-2330 bp) are the most effective in inducing immunity against JEV but not the middle fragment E(A2) (1518-1877 bp) (see for orientation of E(A1), E(A2) and E(B) in E gene). Under the notion that molecular complexity determines the outcome of immune response of the host, E(B) being shorter, simpler in molecular structure and can be easily expressed in soluble form in E. coli (as opposed to insoluble E(A1)), E(B) probably will be the choice as a candidate vaccine to protect the host against JEV infection. PMID- 15748808 TI - Comparative evaluation of apoptosis induced by Shiga toxin 1 and/or lipopolysaccharides in human monocytic and macrophage-like cells. AB - The enteric pathogens Shigella dysenteriae serotype 1 and Shiga toxin-producing Escherichia coli share the property of expressing the structurally and functionally related cytotoxins that comprise the Shiga toxin (Stx) family. Stx producing bacteria are causative agents of bloody diarrheal diseases that may progress to life threatening complications involving the destruction of blood vessels in the kidneys and the central nervous system (CNS). The precise mechanisms of toxin transport across the gut epithelial barrier, and the role of innate immunity in the development of systemic complications, remain to be fully characterized. Earlier studies suggested that Stxs and lipopolysaccharides (LPS) induce the expression of proinflammatory cytokines from differentiated (macrophage-like) THP-1 cells. These cytokines may exacerbate vascular damage by up-regulating the expression of toxin receptors on endothelial cells. Purified Stxs have also been shown to induce apoptosis of epithelial and endothelial cells in vitro, but a comparative evaluation of Stx-induced apoptosis of monocytes and macrophages has not been reported. We used FACS, TUNEL, and DNA laddering analyses to show that Shiga toxin-1 (Stx1) and LPS induce apoptosis in undifferentiated and differentiated THP-1 cells, although the kinetics and extent of apoptosis induction differ between monocytic and macrophage-like cells. Stx1 induced apoptosis is A-subunit-dependent. Stx1 and LPS trigger DNA fragmentation and caspase-3 activation, as evidenced by the cleavage of poly(ADP-ribose) polymerase (PARP). Induction of apoptosis in response to Stx1 and/or LPS treatment occurs without the widespread transcriptional activation of apoptosis related genes. Finally, we present a model of the role of macrophages and monocytes in the pathogenesis of disease caused by Stxs. PMID- 15748809 TI - Comparison of wild type with recombinant Clostridium difficile toxin A. AB - Toxins A and B from Clostridium difficile are single-chain proteins of 308,000 and 270,000 Da, respectively. They possess transferase activity to monoglucosylate proteins of the Rho GTPase family whereby Rho, Rac, and Cdc42 are the canonical substrates. For application of these toxins as specific Rho GTPase inhibitors the highest possible purity is of crucial interest. We, therefore, expressed recombinant His-tagged toxin A using the Bacillus megaterium expression system. Specific antisera raised against the native toxin A from C. difficile and the recombinant toxin, respectively, showed identical sensitivity and specificity in Western blot and ELISA analyses towards both toxins. By comparison of both toxins in functional studies we showed that the recombinant toxin was about two times more cytotoxic than the native toxin, and the glucosyltransferase-activity of the recombinant toxin was even 10-fold increased. However, recombinant toxin A showed one essential difference to the classically purified one. The reported transferase-independent effect of toxin A to release cytochrome c from isolated mitochondria was not exhibited by the recombinant toxin A. This putative mitochondrial effect decreased with increased purity of toxin A, and was absent with recombinant toxin, strongly suggesting an clostridial contamination responsible. In summary, we tested the recombinant toxin A to be at least an adequate substitute for the native toxin, bearing the advantage of a rapid single step purification and the absence of biological active contaminations. PMID- 15748810 TI - Binding of Porphyromonas gingivalis gingipains to human CD4(+) T cells preferentially down-regulates surface CD2 and CD4 with little affect on co stimulatory molecule expression. AB - The role of Porphyromonas gingivalis cysteine proteinases (gingipains) in the evasion of host cell-mediated immunity has not been fully determined. In this study, modulation by gingipains of accessory and co-stimulatory molecule expression on human CD4(+) T cells was evaluated. Arg-gingipain rather than Lys gingipain binds to resting CD4(+) T cells in the presence of serum. The constitutive expression of CD28 on T cells was slightly up-regulated following challenge with gingipains, whereas CD45 and CD3 were not affected. Binding of anti-CD2 and anti-CD4 monoclonal antibodies (mAbs) was reduced after challenge of T cells with gingipains, but restored to 50 and 100%, respectively, of control levels, after 48h of incubation in medium depleted of gingipains. The induced expression, by anti-CD3 mAb, of CTLA-4, CD25, and CD40 ligand (CD40L) was decreased following incubation of T cells with gingipains which also led to decreased response to anti-CD3 and anti-CD28 mAbs as shown by reduction of interleukin-2 (IL-2) production. Cumulatively, these results indicate that activated gingipains attach to T cells and preferentially cleave CD2 and CD4 molecules, with potential to impair T cell responses at periodontal sites. PMID- 15748811 TI - Metalloproteinase-7 contributes to joint destruction in Staphylococcus aureus induced arthritis. AB - Septic arthritis induced by Staphylococcus aureus causes a rapid destruction of joint cartilage and periarticular bone. The mechanisms behind this phenomenon are not fully understood. Earlier studies have shown that cytokines and metalloproteinases are of importance in bone metabolism. Matrix metalloproteinase 7 (MMP-7) has pleiotropic function including facilitating migration of both macrophages and neutrophils. The aim of this study has been to investigate the significance of MMP-7 expression in septic arthritis. MMP-7 deficient mice and congeneic controls were intravenously inoculated with an arthritogenic dose of S. aureus LS-1. This study shows that MMP-7 deficient mice exposed to S. aureus developed significantly less severe arthritis both clinically and histologically. Despite this finding, bacterial growth in the deficient animals was significantly increased. In vitro responses to staphylococcal antigens and superantigens did not differ between MMP-7(+/+) and MMP-7(-/-) mice with respect to cytokine production and if anything increased the production of certain chemokines. In addition MMP-7(-/-) mice exhibited decreased numbers of peripheral blood mononuclear cells before and one day after bacterial inoculation, but increased numbers of peripheral granulocytes on day 1. In conclusion, MMP-7 contributes to the development of a destructive course of septic arthritis despite decreased bacterial load. In addition, expression of MMP-7 is of importance for the distribution of peripheral leukocytes. PMID- 15748812 TI - Burkholderia pseudomallei invasion and activation of epithelial cells requires activation of p38 mitogen-activated protein kinase. AB - Burkholderia pseudomallei is a causative agent of melioidosis. This gram-negative bacterium is able to survive inside the macrophages and also able to invade non phagocytic cells including epithelial cells. Interaction of pathogenic bacteria to the host cells is frequently associated with activation of mitogen-activated protein (MAP) kinases signaling activity. In this study, we demonstrated that B. pseudomallei stimulated p38 MAP kinase of human alveolar lung epithelial cell line (A549). Phosphorylation of p38 was observed after 15 min, attained a maximal level at 60 min after the infection. A specific inhibitor of p38 phosphorylation, SB 203580, was able to inhibit invasion of this bacterium into the cells suggesting that invasion of B. pseudomallei required activation of p38. In contrast, wortmannin which is a specific inhibitor of phosphoinositide 3-kinase (PI3-kinase) failed to inhibit the invasion. Moreover, SB 203580 can also interfere with IkappaBalpha degradation and IL-8 mRNA expression, indicating that the phosphorylation of p38 occurred upstream of NF-kappaB activation. Cytochalasin D, an inhibitor of actin polymerization needed for internalisation of bacteria, did not have any effect on the phosphorylation of p38. These results indicate that B. pseudomallei stimulate phosphorylation of p38 making by initial contact with the cell surface components and do not require internalisation and interaction with intracellular cytoplasmic components of the cells. PMID- 15748814 TI - Apoptotic activity and sub-cellular localization of a T4SS-associated CagE homologue in Actinobacillus actinomycetemcomitans. AB - A potent virulence factor, cagE homologue of Actinobacillus actinomycetemcomitans, was identified via an expression cloning strategy and periodontitis-associated CD4(+)T-cells of a humanized mouse model. Through the immuno-gold labeling with transmission electron microscopy, immunofluorescent staining, in vitro co-cultures and Western blot studies, the resulting data clearly demonstrate that: (i) in CagE-homologue treated human epithelia in vitro, there are ultra-structural features of plasma membrane blebbing, sub-cellular disorganization with condensed and marginalized chromatins along the nuclear membrane, consistent with the pro-apoptotic characteristics, (ii) the disturbed membrane integrity detected above is associated with localization of the CagE proteins on target cell surface, and (iii) CagE-homologue is located in the cytoplasm of A. actinomycetemcomitans and associated with a bacterial type-IV secretion system (T4SS), suggesting that its translocation is required for secretion. Thus, CagE-homologue may be critically involved in A. actinomycetemcomitans-induced tissue destruction, inflammation and subsequent adverse immunity in periodontal pathogenesis. PMID- 15748813 TI - Evaluation of host immune responses to pulmonary cryptococcosis using a temperature-sensitive C. neoformans calcineurin A mutant strain. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that threatens individuals with impaired cell-mediated immunity (CMI). Presently, there are no standardized vaccines available to prevent cryptococcal infections and conventional anti-fungal drug therapy does not induce host immune reactivity and thus cannot efficiently resolve C. neoformans infections in immunocompromised individuals. The present study was designed to characterize pulmonary immune responses following infection with an avirulent temperature-sensitive (ts) mutant, calcineurin A1 (cna1) compared to the pathogenic C. neoformans strain H99 and its potential to induce protective anti-cryptococcal immunity. Host CMI responses in cna1-inoculated mice were observed to be dose-dependent, and comprise increases in pulmonary macrophages and CD4(+) T lymphocytes. However, cytokine analysis demonstrated a mixed pulmonary cytokine response (increases in IL-4, and MCP-1) with no induction of IFN-gamma. Also, pre-immunization with the ts cna1 mutant did not result in protection from a subsequent secondary pulmonary infection with the pathogenic C. neoformans strain H99. Taken together, these results suggest that host pulmonary CMI responses to the ts cna1 mutant that is eventually eliminated from the host without the induction of IFN-gamma appear to be dose-dependent, diverse, and require further stimulation to induce C. neoformans-specific Th1-type cytokine responses to resolve subsequent experimental pulmonary cryptococcal infections. PMID- 15748816 TI - Effect of compatibility on the pharmacokinetic characteristics of ginsenosides. AB - Pharmacokinetic research, which is one of the most important parts of preclinic research, plays an important role in guiding medicine compatibility and preparation improvement. In this paper, the influence of the compatibility of GGV on pharmacokinetics of ginsenosides (GS) was studied, which consisted of ginsenosides, ginkgo biloba extract (GBE) and a chemical monomer V. The result indicated that the addition of either GBE or V could influence the pharmacokinetic parameters of ginsenosides and the influence was different when different administering routes were adopted. Therefore, it could be concluded that there are interactions among GS, GBE and V, and the synergetic and inhibitory effects of the three ingredients contribute to the pharmacological effect of GGV. PMID- 15748815 TI - 'In vivo' studies on the pathophysiological mechanism of Vibrio parahaemolyticus TDH(+)-induced secretion. AB - The thermostable direct haemolysin (TDH) is considered to be the major virulence factors of Vibrio parahaemolyticus; however, poor information is available about its mechanism of action. In our study we examined the capacity of two V. parahaemolyticus TDH-producers (strains 2067 and 3305) to induce fluid secretion in rat ileal loop and to reveal the role of calcium ions (Ca(2+)), calmodulin (CaM), and protein kinase C (PKC) in V. parahaemolyticus TDH(+)-induced fluid secretion. The results show that V. parahaemolyticus TDH(+) strains were able to induce secretion in small intestine; on the contrary, this ability was not evidenced in the V. parahaemolyticus TDH(-) strain used as negative control. The data suggest an enterotoxic activity of haemolysin. Calcium ionophore A23187 and 1-verapamil (calcium channel blocker), when injected alone, induced fluid accumulation in the control loops. A further increase in fluid accumulation (P<0.001) was noted when calcium ionophore was injected along with bacterial suspension of both TDH(+) strains and a significant decrease (P<0.001) in experimental loops when 1-verapamil was inoculated along with bacterial suspension. The other modulating agents increased fluid accumulation in both control and experimental loops, without significant differences with respect to the positive control. Our findings suggest that Ca(2+) appears to be an important messenger involved in the stimulation of intestinal secretion, contrary to PKC and calmodulin which do not appear to have any role. PMID- 15748817 TI - Properties of galactocerebroside layers transferred to glassy carbon electrodes: effect of an applied electric field. AB - Galactocerebroside films deposited onto glassy carbon electrodes have been previously studied through the electrochemical response of a redox couple present in solution. Those experiments indicated that the film is inhomogeneous and that there are lipid-free places. In this work, we present experimental results indicating that those bare regions are formed when the electrode is introduced in an aqueous solution, and that the size and/or amount of uncovered domains increase when negative potentials are applied to the film. The experimental techniques employed for these findings are epifluorescence microscopy and ellipsometry. PMID- 15748818 TI - Surface thermodynamics study of monolayers formed with heteroacid phospholipids of biological interest. AB - The interaction of 1-palmitoy-2-oleoyl-sn-glycero-3-phosphocoline (POPC) and 1 palmitoy-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), two of the major components in biological membranes, were investigated using the monolayer technique at the air-water interface. The pressure-area isotherms indicate that both phospholipids are miscible through all range of compositions. POPE-POPC form stable mixtures, with a minimum for the Gibbs energy of mixing at X(POPC) = 0.4. A virial equation of state was fitted to the experimental values. Positive values found for the second virial coefficient indicate repulsion between POPC and POPE. The interaction parameter was evaluated which indicated that a corresponding decrease in the repulsion occurs when POPC molar fraction is low. This effect suggests the existence of hydrogen bonds between POPE and the water beneath the interface. PMID- 15748819 TI - Na,K-ATPase reconstituted in liposomes: effects of lipid composition on hydrolytic activity and enzyme orientation. AB - In this paper, the reconstitution of Na,K-ATPase in liposomes (formed by single or mixed phospholipids and cholesterol) was investigated and the enzyme orientation was determined on kinetic basis using only specific inhibitors of ATP hydrolysis. A condition of foremost importance for enzyme reconstitution is the achievement of complete solubilization of the lipid in the initial stage of the cosolubilization process for the subsequent formation of the liposomes and/or proteoliposomes. PC-liposomes showed that increasing the fatty acid chain length increases the percentage of Na,K-ATPase incorporated. The average diameter of the proteoliposomes also increases in proportion, reaching a maximum with phospholipids with 16 carbon chains, resulting in 75.1% protein reconstitution and 319.4 nm diameter size, respectively. Binary lipid systems with PC and PE were efficient for incorporation of Na,K-ATPase, depending on the lipid:protein ratio used, varying from 15 to 80% recovery of total ATPase activity. The best results for Na,K-ATPase reconstitution using PC and PE mixture were obtained using a lipid:lipid ratio 1:1 (w/w) and lipid:protein 1:3 (w/w). Integrity studies using calcein release mediated by detergent or alamethicin, in association with inhibition of ATPase activity (ouabain and vanadate) showed that the enzyme is oriented inside-out in DPPC:DPPE proteoliposomes. In these vesicular systems, the enzyme is reconstituted with about 78.9% ATPase activity recovery and 89% protein incorporation, with an average diameter of 140 nm. Systems constituted by DPPC:DPPE, DPPC:DLOPE or DLOPC:DLOPE showed approximately 80, 71 and 70% of recovery of total ATPase activity, but no homogeneity in the distribution of Na,K-ATPase orientation. Reconstitution of Na,K-ATPase in DPPC:DPPE:cholesterol or DPPC:DLOPE:cholesterol systems (55% of cholesterol) showed recovery of about 86 and 82%, respectively, of its total ATPase activity. The results point to an important effect of the lipid acyl chain length and lipid protein ratio in relation to the composition of the lipid matrix to finely tune the structural asymmetry and the amount of enzyme that can be incorporated a lipid bilayer vesicle while preserving membrane permeability. PMID- 15748821 TI - Properties of various phospholipid mixtures as emulsifiers or dispersing agents in nanoparticle drug carrier preparations. AB - The characteristics of mixed phospholipids were examined when used as dispersing agents and emulsifiers. Synthesized phospholipids were mixed to investigate the potential effects of different hydrophilic or lipophilic groups on emulsification and dispersion. To examine the effects of the hydrophilic polar head group on the dispersing or emulsifying potency of phospholipids, l-alpha-phosphatidylcholine dimyristoyl (DMPC) and l-alpha-phosphatidylethanolamine dimyristoyl (DMPE) were mixed in various ratios. Moreover, all combinations of two kinds of phosphatidylcholines (PCs) out of l-alpha-phosphatidylcholine dilauroyl (DLPC), DMPC, l-alpha-phosphatidylcholine dipalmitoyl (DPPC) and l-alpha phosphatidylcholine distearoyl (DSPC) were tested (50:50, w/w) to examine the effects of the hydrophobic carbon chains on the dispersing or emulsifying potency of phospholipids. Mean diameters of vesicles and O/W emulsions prepared by sonication were measured. Vesicles prepared with DMPC-DMPE mixtures gave larger particle sizes than those of DMPC alone. Particle sizes of vesicles prepared with a mixture of two kinds of PCs increased when adding a PC with a longer carbon chain, while particle sizes in a mixture with a PC having a shorter carbon chain was comparable to those in pure PC. In vesicles that were generated by hydration of phospholipids and had a bilayer form, the physical form of the phospholipids consisting of bilayers was thought to be an important factor influencing particle sizes. Among the emulsions, DMPC-DMPE mixtures gave a similar droplet size to DMPC alone. Droplet size in emulsions prepared with a mixture of two kinds of PCs had a strong positive correlation with the total number of carbons, which corresponds to hydrophilic-lipophilic balance (HLB). In O/W emulsions, in which phospholipids were absorbed at water-oil interfaces and which have a single layer form, HLB was thought to be a major factor in the determination of particle size; likewise with non-ionic emulsifiers. PMID- 15748820 TI - Effect of soft segment length on properties of fluorinated polyurethanes. AB - The effects of soft segment length on the variations in morphology, surface composition, and hydrophilicity have been studied in fluorinated polyurethanes (FPUs) and correlated with their preliminary blood compatibility as evidenced by in vitro platelet adhesion experiments. The fluorinated polyurethanes were obtained using hexamethylene diisocyanate (HDI) and chain extender of 2,2,3,3 tetrafluoro-1,4-butanediol (TF) as the hard segment as well as various soft segments-polytetramethyl oxides (PTMO) with molecular weights of 650, 1000, 1400, and 2000. The increased phase separation in hard-segment domains with lengthening soft segment was observed by FT-IR, which is believed to result in enhanced strength of hydrogen bonds and good hard-segment order arrangement. Thin-film XRD results indicate at least three lateral distances existing between adjacent hard segments in the crystallized hard segment. Their distribution depends strongly on the length of soft segment. Lengthening soft segment promotes the formation of dense arrangement of crystallized hard segments. Compared with the effect of phase separation, surface composition was found to exert a major influence on the preliminary blood compatibility of fluorinated polyurethanes. Increasing fluorine content by decreasing soft segment length promotes reduction in platelet adhesion and activation on polyurethane surfaces. PMID- 15748822 TI - Imaging of reconstituted purple membranes by atomic force microscopy. AB - The organization of bacteriorhodopsin (bR) within reconstituted purple membranes (RPM) was examined using atomic force microscopy (AFM). Five reconstituted species were examined: RPM 3 (bR/native polar lipids/dimyristoylphosphatidylcholine (DMPC) in a 1:9:14 molar ratio), RPM 4 (bR/native polar lipids in a 1:7 molar ratio), RPM 5 (bR/native polar lipids/1,2 di-O-phytanyl-sn-glycerol in a 1:3.5:6.1 molar ratio), RPM 6 (bR/native polar lipids/1,2-di-O-phytanyl-sn-glycero-3-phosphocholine in a 1:3.5:4.9 molar ratio), and RPM 7 (bR/native polar lipids/1,2-diphytanoyl-sn-glycero-3-[phospho-L-serine] in a 1:3.5:4.6 molar ratio). RPM 3 patches adsorbed onto mica exhibit domains of crystallized bR trimers arranged in a hexagonal packing structure, similar to those found in native purple membrane (NPM). These domains are enclosed by DMPC rich regions. RPM 4 patches were observed to have larger domains of crystallized bR, with trimer orientation 30 degrees different from that found in NPM. The bR rich domains are enclosed by a large, protein-free, lipid-rich region. The topography of RPM 5 was difficult to resolve as the surface had no discernable patterns or structure. The topographies of RPM 6 and 7 were similar to that found in RPM 3 in that higher domains were formed within the patch adsorbed onto mica. They may contain protein-rich regions, but clear images of protein arrangement could not be obtained using AFM. This may be a result of imaging limitations or of the lack of organization of bR within these domains. PMID- 15748823 TI - Yeast cells long-term interaction with asbestos fibers. AB - Saccharomyces cerevisiae was supported on chrysotile, crocidolite and lixiviated chrysotile. Samples of the supported cells and free cells were observed by confocal laser scanning microscopy. After 30 days, the free cells showed no viability when stored at 30 degrees C, and a viability of 40% when stored at 4 degrees C. Supported cells stored at 30 degrees C were more viable than the free cells at early times, but showed no viability after 30 days. Samples stored at 4 degrees C showed that the adhered cells are more viable than the free cells, up to 30 days. Cells supported on chrysotile and lixiviated chrysotile had 80% viability, and on crocidolite 70% viability. Scanning electron microscopy showed that cells supported on lixiviated chrysotile are fully covered by the support, but crocidolite fibers adhere less, since they are stiffer. Fermentation experiments performed after 3 years storage showed that four from the five lixiviated chrysotile samples and one of the three crocidolite samples were active. In all cases, a delay time for the onset of fermentation was observed indicating a state of latency. PMID- 15748824 TI - Properties of two-component Langmuir monolayer of single chain perfluorinated carboxylic acids with dipalmitoylphosphatidylcholine (DPPC). AB - The surface pressure (pi)- and the surface potential (DeltaV)-area (A) isotherms were obtained for two-component monolayers of four different perfluorocarboxylic acids (FCns; perfluorododecanoic acid: FC12, perfluorotetradecanoic acid: FC14, perfluorohexadecanoic acid: FC16, perfluorooctadecanoic acid: FC18) with dipalmitoylphosphatidylcholine (DPPC) on substrate solution of 0.15 M NaCl (pH 2.0) at 298.2 K as a function of compositions in the mixtures by employing the Wilhelmy method, the ionizing electrode method, the fluorescence microscopy, and the atomic force microscopy. The data for the two-component monolayers on these systems were analyzed in terms of the additivity rule. Assuming a regular surface mixture, the Joos equation which allows one to describe the collapse pressure of a two-component monolayer with miscible components was used to declare the miscibility of the monolayer state, and an interaction parameter and an interaction energy were calculated. The new finding was that FCns and DPPC are miscible or immiscible depending on chain length increment of fluorocarbon part. That is, FC12/DPPC monolayer was perfectly miscible, and FC14/DPPC, and FC16/DPPC (0 < or = X(FC16) < or = 0.3) monolayers were partially miscible. While FC16/DPPC (0.3 < X(FC16) < 1) and FC18/DPPC systems are immiscible in the monolayer state. Furthermore, the mean molecular area, the surface dipole moment, and the phase diagrams enabled us to estimate the molecular orientation of four different perfluorocarboxylic acids/DPPC in the two-component monolayer state. One type of phase diagrams was obtained and classified into the positive azeotropic type. The miscibility of FCns and DPPC in the monolayer was also supported by fluorescence microscopy and atomic force microscopy. FC12/DPPC, FC14/DPPC and FC16/DPPC (0 < or = X(FC16) < or = 0.3) two-component monolayers on 0.15 M NaCl (pH 2) showed that FC12, FC14 and FC16 (0 < or = X(FC16) < or = 0.3) can dissolve or partially dissolve the ordered solid DPPC domains formed upon compression. This indicates that these fluorinated amphiphiles soften or harden the lipid depending on their chain length. PMID- 15748825 TI - The fabrication of a colloidal gold-carbon nanotubes composite film on a gold electrode and its application for the determination of cytochrome c. AB - Colloid Au (Au(nano)) with a diameter of about 20 nm was prepared and used in combination with the multi-wall carbon nanotubes (MWNTs) to modify a gold electrode. Dihexadecylphosphate (DHP) dispersed in Au(nano) aqueous solution was used to solubilize MWNTs. Deposition of Au(nano) on MWNTs was realized as illustrated by TEM micrographs. The DHP formed a network that connected Au(nano) and MWNTs to the gold electrode surface. The Au(nano)-MWNTs-DHP composite film on the gold electrode surface was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammmetry (CV). The composite film modified gold electrode was used to detect cytochrome c and a pair of well-defined redox waves was obtained. It was found that the composite film promoted the redox of horse heart cytochrome c and its effect was developed for the determination of cytochrome c. PMID- 15748826 TI - Dialkylphosphatidylcholine and egg yolk lecithin for emulsification of various triglycerides. AB - Synthesized saturated phosphatidylcholine (PC) and egg yolk lecithin (EYL) were investigated to explore their influence on particle sizes in emulsions when dispersing various triglycerides (TG). One of four different kinds of synthesized saturated PC (DLPC, DMPC, DPPC and DSPC) or three different kinds of EYL (purified EYL (PEL) and hydrogenated purified EYL with two different iodine values (IV), R-20 and R-5), 2.5% (w/w) glycerol solution and one of four kinds of TG (tricaprylin, tricaprin, trilaurin and trimyristin) were sonicated five times for 1 min with intervals of 0.5 min. When using four kinds of synthesized saturated PCs as emulsifiers, the carbon numbers of each PC had a strong correlation with the mean diameters of the emulsion when analyzed with each of the four kinds of TG used in the study (regression function ranged from 0.811 to 0.915). The carbon numbers of the TG had less correlation with the mean diameters than the PC in simple regression analysis (regression function ranged from 0.236 to 0.875). Multiple regression analysis using the carbon numbers both of the PC and TG as independent variables was remarkably significant in the regression function (2.0 x 10(-14)) and all regression coefficients (2.7 x 10(-13), 5.8 x 10(-7) and 1.9 x 10(-9) for PC, TG and intercept, respectively). Among the regression coefficients, the contribution of the carbon number of the PC was the most significant. These results indicated that a multiple regression function should be useful to estimate the mean diameters of emulsion droplets in any combinations of PC and TG used in this study. In the experiments using three kinds of EYL, the mean diameters also tended to increase according to the order of PEL, R-20 and R-5, which corresponds to the order of degrees of saturation (IV = 75, 20 and 2, respectively). The experimental values for EYL were compared with the estimated values calculated by the multiple regression function derived from synthesized PC data using the arithmetic carbon number, based on the components of each EYL. The estimated mean diameters were at comparable levels to the corresponding experimental mean diameters in the most saturated hydrogenated lecithin (R-5), while those were larger than the experimental mean diameters in two less saturated kinds of lecithin (R-20 and purified EYL). These findings gave useful information on the mean diameters of emulsion droplets when designing an emulsion formulation using a particular combination of a phospholipid and triglyceride. PMID- 15748827 TI - Effect of nomegestrol acetate on estrogen biosynthesis and transformation in MCF 7 and T47-D breast cancer cells. AB - Although ovaries serve as the primary source of estrogen for pre-menopausal women, after menopause estrogen biosynthesis from circulating precursors occurs in peripheral tissues by the action of several enzymes, 17beta-hydroxysteroid dehydrogenase 1 (17beta-HSD1), aromatase and estrogen sulfatase. In the breast, both normal and tumoral tissues have been shown to be capable of synthesizing estrogens, and this local estrogen production can be implicated in the development of breast tumors. In these tissues, estradiol (E(2)) can be synthesized by three pathways: (1) estrone sulfatase transforms estrogen sulfates into bioactive estrogens, (2) 17beta-HSD1 converts estrone (E(1)) into E(2), (3) aromatase which converts androgens into estrogens is also present and contributes to the in situ synthesis of active estrogens but to a far lesser extent than estrone sulfatase. Quantitative assessment of E(2) formation in human breast tumors indicates that metabolism of estrone sulfate (E(1)S) via the sulfatase pathway produces 100-500 times more E(2) than androgen aromatization. Breast tissue also possesses the estrogen sulfotransferase involved in the conversion of estrogens into their sulfates that are biologically inactive. In the present review, we summarized the action of the 19-nor-progestin nomegestrol acetate (NOMAC) on the sulfatase, 17beta-HSD1 and sulfotransferase activities in the hormone-dependent MCF-7 and T47-D human breast cancer cell lines. Using physiological doses of substrates NOMAC blocks very significantly the conversion of E(1)S to E(2). It inhibits the transformation of E(1) to E(2). NOMAC has a stimulatory effect on sulfotransferase activity in both cell lines, with a strong stimulating effect at low doses but only a weak effect at high concentrations. The effects on the three enzymes are always stronger in the progesterone-receptor rich T47-D cell line as compared with the MCF-7 cell line. Besides, no effect is found for NOMAC on the transformation of androstenedione to E(1) in the aromatase rich choriocarcinoma cell line JEG-3. In conclusion, the inhibitory effect provoked by NOMAC on the enzymes involved in the biosynthesis of E(2) (sulfatase and 17HSD pathways) in estrogen-dependent breast cancer, as well as the stimulatory effect on the formation of the inactive E(1)S, can open attractive perspectives for future clinical trials. PMID- 15748828 TI - Aromatase expression in ovarian epithelial cancers. AB - Our study focused on aromatase cytochrome P450 (CYP19) expression in ovarian epithelial normal and cancer cells and tissues. Aromatase mRNA expression was analyzed by real-time PCR in ovarian epithelial cancer cell lines, in human ovarian surface epithelial (HOSE) cell primary cultures, and in ovarian tissue specimens (n=94), including normal ovaries, ovarian cysts and cancers. Aromatase mRNA was found to be expressed in HOSE cells, in BG1, PEO4 and PEO14, but not in SKOV3 and NIH:OVCAR-3 ovarian cancer cell lines. Correlation analysis of aromatase expression was performed according to clinical, histological and biological parameters. Aromatase expression in ovarian tissue specimens was higher in normal ovaries and cysts than in cancers (P<0.0001). Using laser capture microdissection in normal postmenopausal ovaries, aromatase was found to be predominantly expressed in epithelial cells as compared to stromal component. Using immunohistochemistry (IHC), aromatase was also detected in the epithelium component. There was an inverse correlation between aromatase and ERalpha expression in ovarian tissues (P<0.001, r=-0.34). In the cancer group, no significant differences in aromatase expression were observed according to tumor histotype, grade, stage and survival. Aromatase activity was evaluated in ovarian epithelial cancer (OEC) cell lines by the tritiated water assay and the effects of third-generation aromatase inhibitors (AIs) on aromatase activity and growth were studied. Letrozole and exemestane were able to completely inhibit aromatase activity in BG1 and PEO14 cell lines. Interestingly, both AI showed an antiproliferative effect on the estrogen responsive BG1 cell line co-expressing aromatase and ERalpha. Aromatase expression was found in ovarian epithelial normal tissues and in some ovarian epithelial cancer cells and tissues. This finding raises the possibility that some tumors may respond to estrogen and provides a basis for ascertaining an antimitogenic effect of AI in a subgroup of ovarian epithelial cancers. PMID- 15748829 TI - Endogenous aromatization of testosterone results in growth stimulation of the human MCF-7 breast cancer cell line. AB - Estrogens produced within breast tumors may play a pivotal role in growth stimulation of the breast cancer cells. However, it is elusive whether the epithelial breast cancer cells themselves synthesize estrogens, or whether the surrounding tumor stromal cells synthesize and supply the cancer cells with estrogen. The aromatase enzyme catalyzes the estrogen production, aromatizing circulating androgens into estrogens. The aim of this study was to investigate aromatase expression and function in a model system of human breast cancer, using the estrogen responsive human MCF-7 breast cancer cell line. Cells were cultured in a low estrogen milieu and treated with estrogens, aromatizable androgens or non-aromatizable androgens. Cell proliferation, expression of estrogen-regulated proteins and aromatase activity were investigated. The MCF-7 cell line was observed to express sufficient aromatase enzyme activity in order to aromatize the androgen testosterone, resulting in a significant cell growth stimulation. The testosterone-mediated growth effect was completely inhibited by the aromatase inhibitors letrozole and 4-hydroxy-androstenedione. Expression studies of estrogen-regulated proteins confirmed that testosterone was aromatized to estrogen in the MCF-7 cells. Thus, the results indicate that epithelial breast cancer cells possess the ability to aromatize circulating androgens to estrogens. PMID- 15748830 TI - Androgen receptor content following heavy resistance exercise in men. AB - The purpose of the present investigation was to examine androgen receptor (AR) content in the vastus lateralis following two resistance exercise protocols of different volume. Nine resistance-trained men (age=24.3+/-4.4 years) performed the squat exercise for 1 (SS) and 6 sets (MS) of 10 repetitions in a random, counter-balanced order. Muscle biopsies were performed at baseline, and 1h following each protocol. Blood was collected prior to, immediately following (IP), and every 15 min after each protocol for 1h. No acute elevations in serum total testosterone were observed following SS, whereas significant 16-23% elevations were observed at IP, 15, and 30 min post-exercise following MS. No acute elevations in plasma cortisol were observed following SS, whereas significant 31-49% elevations were observed for MS at IP, 15, and 30 min post exercise. Androgen receptor content did not change 1h following SS but significantly decreased by 46% following MS. These results demonstrated that a higher volume of resistance exercise resulted in down-regulation of AR content 1h post-exercise. This may have been due to greater protein catabolism associated with the higher level of stress following higher-volume resistance exercise. PMID- 15748831 TI - Effects of chronic anabolic steroid treatment on tonic and reflex cardiovascular control in male rats. AB - The aim of this study was to analyze the cardiovascular effects of chronic stanozolol administration in male rats. The rats were randomly assigned to one of three groups: (1) control (n=12), (2) chronic treatment with low dose of stanozolol (LD, n=18, 5 mg/kgweek) and; (3) treatment with high dose of stanozolol (HD, n=28, 20 mg/kgweek). Mean arterial pressure (MAP) was higher in both HD (128+/-2.2 mmHg) and LD (126+/-2.5 mmHg) than control (116+/-2 mmHg). The LD group showed an increase in cardiac output (control 121+/-2.5, LD 154+/-5.9 ml/min), whereas in the HD group total peripheral resistance increased (control 1.03+/-0.07, HD 1.26+/-0.07 mmHg/ml/min). Acute sympathetic blockade caused a similar decrease in MAP in all groups. In conscious rats, the baroreflex index for bradycardia (control -3.7+/-0.4, LD -2.0+/-0.1 beat/mmHg) and tachycardia (control -3.6+/-0.3, LD -4.7+/-0.2 beat/mmHg) responses changed only in the LD group. Cardiac hypertrophy was observed in both treated groups (P<0.05). In conclusion, hypertension with differential hemodynamic changes and alterations in the reflex control in heart rate is seen at different stanozolol doses, which may be important variables in the cardiovascular effects of anabolic steroids. PMID- 15748832 TI - Localization of type 7 17beta-hydroxysteroid dehydrogenase in mouse tissues. In situ hybridization studies. AB - The enzyme type 7 17beta-hydroxysteroid dehydrogenase (17beta-HSD) selectively catalyzes the conversion of estrone (E1) into estradiol (E2). In order to obtain detailed information about the exact sites of action of type 7 17beta-HSD, we have studied the cellular localization of type 7 17beta-HSD mRNA in mouse tissues using in situ hybridization (ISH). In parallel studies, we also measured the enzyme mRNA levels by quantitative real time (RT)-PCR. In the ovary, strong hybridization signal was restricted to corpus luteum cells. In the female mammary gland, type 7 17beta-HSD mRNA was found to be expressed in stromal cells surrounding the ducts. In the clitoral and preputial glands, specific labeling was observed in the epithelial cells of both acini and small ducts. In the adrenal gland, hybridization signal was observed in the zona fasciculata and reticularis in the cortex. In the liver, hybridization signal was found in all the hepatocytes. In the colon, type 7 17beta-HSD mRNA expression was restricted to epithelial cells of the mucosa. From the results obtained with quantitative real time RT-PCR, it appears, with a very few exceptions, that in tissues exhibiting low mRNA expression no ISH signal could be detected. The present data suggest that E2 can be formed through the action of type 7 17beta-HSD in specific cell types in the ovary and peripheral tissues, in addition to type 1 17beta-HSD, thus providing tissues with an alternative route of formation of E2. PMID- 15748833 TI - Sex hormone regulation of anti-bacterial activity in rat uterine secretions and apical release of anti-bacterial factor(s) by uterine epithelial cells in culture. AB - In mature female rats, sex hormones regulate the reproductive (estrous) cycle to optimize mating and fertility. During the part of the estrous cycle when mating occurs, and when estrogen is the dominant sex hormone, the uterus is susceptible to infection with bacteria that can be deleterious for survival and fertility. The present study investigated whether sex hormones regulate innate immunity in the female reproductive tract by affecting the secretion of an anti-bacterial factor(s) in the rat uterus. Uterine fluids from intact rats at the proestrous stage of the estrous cycle significantly inhibited Staphylococcus aureus growth. When ovariectomized rats were treated with estradiol, anti-bacterial activity against both S. aureus and Escherichia coli increased in uterine secretions with hormone treatment. In contrast, rats injected with either progesterone and estradiol or progesterone alone displayed no bactericidal activity indicating that progesterone reversed the stimulatory effect of estradiol on anti-bacterial activity. In other studies, isolated uterine epithelial cells from intact animals were grown to confluence and high transepithelial resistance on cell inserts. Analysis of apical secretions indicated that a soluble factor(s) is released by polarized epithelial cells which inhibits bacterial growth. These results demonstrate that sex hormones influence the presence of a broad-spectrum bactericidal factor(s) in luminal secretions of the rat uterus. Further these studies suggest that epithelial cells which line the uterine lumen are a primary source of anti-bacterial activity. PMID- 15748834 TI - Testicular adrenal rest tumours in salt wasting congenital adrenal hyperplasia (in vivo and in vitro studies). AB - We describe the case of a 20-year-old patient with salt-wasting congenital adrenal hyperplasia (CAH) related to 21-hydroxylase deficiency. Bilateral craggy testicular tumours were found, requiring histological evaluation. Prior to the surgical procedure, the patient was treated with dexamethasone (he presented cortisol deficiency) and was stimulated with ACTH. High levels of 11beta-OH steroids measured in the gonadal vein, compared with peripheral blood samples suggested the presence of adrenal rests. Incubation of the tumours (which could not be differentiated histologically, from Leydig tissue), with radioactive steroid precursors was carried out. The results revealed the testicular tumours were of adrenal tissue origin, associated with 21-hydroxylase deficiency. The patient's non-compliance to glucocorticoid treatment was the main cause of his hypogonadotropic hypogonadism. PMID- 15748835 TI - Inhibitory effects of flavonoids on the reduction of progesterone to 20alpha hydroxyprogesterone in rat liver. AB - The first aim of this study is to characterize the reduction of progesterone in rat liver. Progesterone was mainly reduced to 20alpha-hydroxyprogesterone in the cytosolic fraction of rat liver. The amount of 20alpha-hydroxyprogesterone formed from progesterone in the cytosolic fraction was significantly larger in the males than in the females and this enzyme reaction proceeded not only in the presence of NADPH, but also in the presence of NADH. Furthermore, we attempted to evaluate the inhibitory effects of 15 flavonoids on the NADPH-dependent reduction of progesterone to 20alpha-hydroxyprogesterone in liver cytosol of male rats. The order of the inhibitory potencies was luteolin>apigenin>quercetin>myricetin=fisetin=kaempferol. Other flavonoids exhibited lower inhibitory potencies. Energy-minimized molecular models demonstrated that a planar benzopyrone ring (A and C rings) with a coplanar phenyl ring (B ring) is a structural characteristic determining the inhibitory effects of flavonoids other than isoflavones. PMID- 15748836 TI - Metabolism of dehydroepiandrosterone by rodent brain cell lines: relationship between 7-hydroxylation and aromatization. AB - The rate of aromatization of 4-androstenedione (AD) and 7-hydroxylation of dehydroepiandrosterone (DHEA) by different neuronal cell lines from fetal rat and mouse brain was compared to that of embryonic rat hippocampal cells in primary culture. The (3)H-labeled steroids were incubated with the cells and the metabolites extracted and separated by thin layer chromatography (TLC), as well as analyzed by high-performance liquid chromatography (HPLC) for further identification. All cell types produced estrone (E(1)) and estradiol (E(2)) from [(3)H]AD but the rate of aromatization was lowest with the rat hippocampal cells in primary culture. With [(3)H]DHEA, BHc.2 mouse hippocampal cells and E(t)C.1 neurons behaved like the mixed cells from rat hippocampus, forming 7-hydroxy DHEA as the almost exclusive product. In contrast, mouse brain BV2 microglia were virtually unable to hydroxylate DHEA at C-7 and yielded estrogen and more testosterone (T) than other cell types tested. These experiments highlight the pivotal role of 3beta-hydroxysteroid dehydrogenase/ketoisomerase in the control of AD formation for its subsequent aromatization to estrogen. It raises the possibility that differences in metabolism of DHEA by certain brain cells could account for differences in their immunomodulatory and neuroprotective functions. Some could exert their effects by converting DHEA to its 7-hydroxylated form while others, like BV2 microglia, by converting DHEA primarily to other C-19 steroids and to estrogen by way of AD. PMID- 15748837 TI - Homing in on intracellular Abeta? AB - In this issue of Neuron, a study by Billings et al. points to intracellular Abeta as a possible cause of neuronal dysfunction. In a mouse model of Alzheimer's disease, Billings et al. link appearance of intraneuronal Abeta to cognitive impairments and then show that "clearance" of intraneuronal Abeta by anti-Abeta antibodies restores cognitive deficits. PMID- 15748838 TI - Courting a cure for fragile X. AB - Fragile X syndrome is the most common heritable cause of mental retardation. Previous work has suggested that overactive signaling by group I metabotropic glutamate receptors (mGluRs) may be a mechanism underlying many of the disease symptoms. As a test of this theory, McBride et al. show that in a Drosophila model for Fragile X syndrome, treatment with mGluR antagonists can rescue short term memory, courtship, and mushroom body defects. PMID- 15748839 TI - Turn the other cheek: viewpoint aftereffects for faces and objects. AB - In this issue of Neuron, Fang and He demonstrate that viewing a face or object from a particular viewpoint leads to an aftereffect whereby later neutral views of the same stimulus appear to be biased away from the initial viewpoint. This new aftereffect suggests that the human brain contains populations of neurons tuned to the angle from which an object is viewed. PMID- 15748840 TI - Unmanageable motivation in addiction: a pathology in prefrontal-accumbens glutamate transmission. AB - Prime diagnostic criteria for drug addiction include uncontrollable urges to obtain drugs and reduced behavioral responding for natural rewards. Cellular adaptations in the glutamate projection from the prefrontal cortex (PFC) to the nucleus accumbens have been discovered in rats withdrawn from cocaine that may underlie these cardinal features of addiction. A hypothesis is articulated that altered G protein signaling in the PFC focuses behavior on drug-associated stimuli, while dysregulated PFC-accumbens synaptic glutamate transmission underlies the unmanageable motivation to seek drugs. PMID- 15748841 TI - Deep brain stimulation for treatment-resistant depression. AB - Treatment-resistant depression is a severely disabling disorder with no proven treatment options once multiple medications, psychotherapy, and electroconvulsive therapy have failed. Based on our preliminary observation that the subgenual cingulate region (Brodmann area 25) is metabolically overactive in treatment resistant depression, we studied whether the application of chronic deep brain stimulation to modulate BA25 could reduce this elevated activity and produce clinical benefit in six patients with refractory depression. Chronic stimulation of white matter tracts adjacent to the subgenual cingulate gyrus was associated with a striking and sustained remission of depression in four of six patients. Antidepressant effects were associated with a marked reduction in local cerebral blood flow as well as changes in downstream limbic and cortical sites, measured using positron emission tomography. These results suggest that disrupting focal pathological activity in limbic-cortical circuits using electrical stimulation of the subgenual cingulate white matter can effectively reverse symptoms in otherwise treatment-resistant depression. PMID- 15748842 TI - Coexpression of two functional odor receptors in one neuron. AB - One of the most fundamental tenets in the field of olfaction is that each olfactory receptor neuron (ORN) expresses a single odorant receptor. However, the one receptor-one neuron principle is difficult to establish rigorously. Here we construct a receptor-to-neuron map for an entire olfactory organ in Drosophila and find that two receptor genes are coexpressed in one class of ORN. Both receptors are functional in an in vivo expression system, they are only 16% identical in amino acid sequence, and the genes that encode them are unlinked. Most importantly, their coexpression has been conserved for >45 million years. Expression of multiple odor receptors in a cell provides an additional degree of freedom for odor coding. PMID- 15748843 TI - Differential encoding of behavior and spatial context in deep and superficial layers of the neocortex. AB - Rodent hippocampal activity is correlated with spatial and behavioral context, but how context affects coding in association neocortex is not well understood. The cellular distribution of the neural activity-regulated immediate-early gene Arc was used to monitor the activity history of cells in CA1, and in deep and superficial layers of posterior parietal and gustatory cortices (which encode movement and taste, respectively), during two behavioral epochs in which spatial and behavioral context were independently manipulated while gustatory input was held constant. Under conditions in which the hippocampus strongly differentiated behavioral and spatial contexts, deep parietal and gustatory layers did not discriminate between spatial contexts, whereas superficial layers in both neocortical regions discriminated well. Deep parietal cells discriminated behavioral context, whereas deep gustatory cortex neurons encoded the two conditions identically. Increased context sensitivity of superficial neocortical layers, which receive more hippocampal outflow, may reflect a general principle of neocortical organization for memory retrieval. PMID- 15748844 TI - Intraneuronal Abeta causes the onset of early Alzheimer's disease-related cognitive deficits in transgenic mice. AB - Progressive memory loss and cognitive dysfunction are the hallmark clinical features of Alzheimer's disease (AD). Identifying the molecular triggers for the onset of AD-related cognitive decline presently requires the use of suitable animal models, such as the 3xTg-AD mice, which develop both amyloid and tangle pathology. Here, we characterize the onset of learning and memory deficits in this model. We report that 2-month-old, prepathologic mice are cognitively unimpaired. The earliest cognitive impairment manifests at 4 months as a deficit in long-term retention and correlates with the accumulation of intraneuronal Abeta in the hippocampus and amygdala. Plaque or tangle pathology is not apparent at this age, suggesting that they contribute to cognitive dysfunction at later time points. Clearance of the intraneuronal Abeta pathology by immunotherapy rescues the early cognitive deficits on a hippocampal-dependent task. Reemergence of the Abeta pathology again leads to cognitive deficits. This study strongly implicates intraneuronal Abeta in the onset of cognitive dysfunction. PMID- 15748845 TI - Regulators acting in combinatorial codes also act independently in single differentiating neurons. AB - In the Drosophila ventral nerve cord, a small number of neurons express the LIM homeodomain gene apterous (ap). These ap neurons can be subdivided based upon axon pathfinding and their expression of neuropeptidergic markers. ap, the zinc finger gene squeeze, the bHLH gene dimmed, and the BMP pathway are all required for proper specification of these cells. Here, using several ap neuron terminal differentiation markers, we have resolved how each of these factors contributes to ap neuron diversity. We find that these factors interact genetically and biochemically in subtype-specific combinatorial codes to determine certain defining aspects of ap neuron subtype identity. However, we also find that ap, dimmed, and squeeze additionally act independently of one another to specify certain other defining aspects of ap neuron subtype identity. Therefore, within single neurons, we show that single regulators acting in numerous molecular contexts differentially specify multiple subtype-specific traits. PMID- 15748846 TI - GDNF and GFRalpha1 promote differentiation and tangential migration of cortical GABAergic neurons. AB - Cortical GABAergic neurons are generated in the ventral telencephalon and migrate dorsally into the cortex following a tangential path. GDNF signaling via GFRalpha1 was found to promote the differentiation of ventral precursors into GABAergic cells, enhancing their neuronal morphology and motility. GDNF stimulated axonal growth in cortical GABAergic neurons and acted as a potent chemoattractant of GABAergic cells. These effects required GFRalpha1 but neither RET nor NCAM, the two transmembrane signaling receptors known for GDNF. Mutant mice lacking GDNF or GFRalpha1, but neither RET nor NCAM, showed reduced numbers of GABAergic cells in the cerebral cortex and hippocampus. We conclude that one of the normal functions of GDNF signaling via GFRalpha1 in the developing brain is to promote the differentiation and migration of cortical GABAergic neurons. The lack of involvement of RET or NCAM in these processes suggests the existence of additional transmembrane effectors for GDNF. PMID- 15748847 TI - Vimentin-dependent spatial translocation of an activated MAP kinase in injured nerve. AB - How are phosphorylated kinases transported over long intracellular distances, such as in the case of axon to cell body signaling after nerve injury? Here, we show that the MAP kinases Erk1 and Erk2 are phosphorylated in sciatic nerve axoplasm upon nerve injury, concomitantly with the production of soluble forms of the intermediate filament vimentin by local translation and calpain cleavage in axoplasm. Vimentin binds phosphorylated Erks (pErk), thus linking pErk to the dynein retrograde motor via direct binding of vimentin to importin beta. Injury induced Elk1 activation and neuronal regeneration are inhibited or delayed in dorsal root ganglion neurons from vimentin null mice, and in rats treated with a MEK inhibitor or with a peptide that prevents pErk-vimentin binding. Thus, soluble vimentin enables spatial translocation of pErk by importins and dynein in lesioned nerve. PMID- 15748848 TI - The beta subunit determines the ligand binding properties of synaptic glycine receptors. AB - Inhibitory glycine receptors (GlyRs) regulate motor coordination and sensory signal processing in spinal cord and other brain regions. GlyRs are pentameric proteins composed of membrane-spanning alpha and beta subunits. Here, site directed mutagenesis combined with homology modeling based on the crystal structure of the acetylcholine binding protein identified key ligand binding residues of recombinant homooligomeric alpha1 and heterooligomeric alpha1beta GlyRs. This disclosed two highly conserved, oppositely charged residues located on adjacent subunit interfaces as being crucial for agonist binding. In addition, the beta subunit was found to determine the ligand binding properties of heterooligomeric GlyRs. Expression of an alpha1beta tandem construct and affinity purification of metabolically labeled GlyRs confirmed a subunit stoichiometry of 2alpha3beta. Because the beta subunit anchors GlyRs at synaptic sites, our results have important implications for the biosynthesis, clustering, and pharmacology of synaptic GlyRs. PMID- 15748849 TI - Generation of silent synapses by acute in vivo expression of CaMKIV and CREB. AB - The transcription factor CREB is critical for several forms of experience dependent plasticity in a range of species and is commonly activated in neurons by calcium/calmodulin-dependent protein kinase IV (CaMKIV). Surprisingly, little is known about the neural circuit adaptations caused by activation of CaMKIV and CREB. Here, we use viral-mediated gene transfer in vivo to examine the consequences of acute expression of constitutively active forms of CaMKIV and CREB on synaptic function in the rodent hippocampus. Acute expression of active CaMKIV or CREB caused an enhancement of both NMDA receptor-mediated synaptic responses and long-term potentiation (LTP). This was accompanied by electrophysiological and morphological changes consistent with the generation of "silent synapses," which provide an ideal substrate for further experience dependent modifications of neural circuitry and which may also be important for the consolidation of long-term synaptic plasticity and memories. PMID- 15748850 TI - Pharmacological rescue of synaptic plasticity, courtship behavior, and mushroom body defects in a Drosophila model of fragile X syndrome. AB - Fragile X syndrome is a leading heritable cause of mental retardation that results from the loss of FMR1 gene function. A Drosophila model for Fragile X syndrome, based on the loss of dfmr1 activity, exhibits phenotypes that bear similarity to Fragile X-related symptoms. Herein, we demonstrate that treatment with metabotropic glutamate receptor (mGluR) antagonists or lithium can rescue courtship and mushroom body defects observed in these flies. Furthermore, we demonstrate that dfmr1 mutants display cognitive deficits in experience-dependent modification of courtship behavior, and treatment with mGluR antagonists or lithium restores these memory defects. These findings implicate enhanced mGluR signaling as the underlying cause of the cognitive, as well as some of the behavioral and neuronal, phenotypes observed in the Drosophila Fragile X model. They also raise the possibility that compounds having similar effects on metabotropic glutamate receptors may ameliorate cognitive and behavioral defects observed in Fragile X patients. PMID- 15748851 TI - Dopaminergic stimulation of local protein synthesis enhances surface expression of GluR1 and synaptic transmission in hippocampal neurons. AB - The use-dependent modification of synapses is strongly influenced by dopamine, a transmitter that participates in both the physiology and pathophysiology of animal behavior. In the hippocampus, dopaminergic signaling is thought to play a key role in protein synthesis-dependent forms of synaptic plasticity. The molecular mechanisms by which dopamine influences synaptic function, however, are not well understood. Using a GFP-based reporter, as well as a small-molecule reporter of endogenous protein synthesis, we show that dopamine D1/D5 receptor activation stimulates local protein synthesis in the dendrites of hippocampal neurons. We also identify the GluR1 subunit of AMPA receptors as one protein upregulated by dopamine receptor activation, with increased incorporation of surface GluR1 at synaptic sites. The insertion of new GluRs is accompanied by an increase in the frequency of miniature synaptic events. Together, these data suggest a local protein synthesis-dependent activation of previously silent synapses as a result of dopamine receptor stimulation. PMID- 15748852 TI - Spatial structure of complex cell receptive fields measured with natural images. AB - Neuronal receptive fields (RFs) play crucial roles in visual processing. While the linear RFs of early neurons have been well studied, RFs of cortical complex cells are nonlinear and therefore difficult to characterize, especially in the context of natural stimuli. In this study, we used a nonlinear technique to compute the RFs of complex cells from their responses to natural images. We found that each RF is well described by a small number of subunits, which are oriented, localized, and bandpass. These subunits contribute to neuronal responses in a contrast-dependent, polarity-invariant manner, and they can largely predict the orientation and spatial frequency tuning of the cell. Although the RF structures measured with natural images were similar to those measured with random stimuli, natural images were more effective for driving complex cells, thus facilitating rapid identification of the subunits. The subunit RF model provides a useful basis for understanding cortical processing of natural stimuli. PMID- 15748853 TI - Viewer-centered object representation in the human visual system revealed by viewpoint aftereffects. AB - Are there neurons representing specific views of objects in the human visual system? A visual selective adaptation method was used to address this question. After visual adaptation to an object viewed either 15 or 30 degrees from one side, when the same object was subsequently presented near the frontal view, the perceived viewing directions were biased in a direction opposite to that of the adapted viewpoint. This aftereffect can be obtained with spatially nonoverlapping adapting and test stimuli, and it depends on the global representation of the adapting stimuli. Viewpoint aftereffects were found within, but not across, categories of objects tested (faces, cars, wire-like objects). The magnitude of this aftereffect depends on the angular difference between the adapting and test viewing angles and grows with increasing duration of adaptation. These results support the existence of object-selective neurons tuned to specific viewing angles in the human visual system. PMID- 15748854 TI - Neural correlates of reaching decisions in dorsal premotor cortex: specification of multiple direction choices and final selection of action. AB - We show that while a primate chooses between two reaching actions, its motor system first represents both options and later reflects selection between them. When two potential targets appeared, many (43%) task-related, directionally tuned cells in dorsal premotor cortex (PMd) discharged if one of the targets was near their preferred direction. At the population level, this generated two simultaneous sustained directional signals corresponding to the current reach options. After a subsequent nonspatial cue identified the correct target, the corresponding directional signal increased, and the signal for the rejected target was suppressed. The PMd population reliably predicted the monkey's response choice, including errors. This supports a planning model in which multiple reach options are initially specified and then gradually eliminated in a competition for overt execution, as more information accumulates. PMID- 15748855 TI - Digestive morphology and enzyme activity in the Andean toad Bufo spinulosus: hard wired or flexible physiology? AB - Gut plasticity is a trait with implications on animal performance. However, and despite their importance as study models in physiology, research on gut flexibility in amphibians is scarce. In the present work, we analyse digestive adjustments of Bufo spinulosus adult individuals to cope with changes in diet quality and quantity at two organizational levels (i.e., digestive morphology and enzymes). We found that changes in gut size are related to the amount of food ingested, but not to diet composition. This is in agreement with "the gut seasonal change" hypothesis and offers a proximal explanation for this change. Digestive enzymatic activity (maltase and aminopeptidase-N) did not change with diet quality or quantity, which agrees with the hypothesis of "hard-wired physiology in adult amphibians". Both hypotheses are in agreement with the general theoretical framework of gut phenotypic flexibility when interpreted in light of amphibian natural history. In addition, our results indicate that the correlation between feeding frequency and the level of gut up-regulation proposed for interspecific comparisons may also be found at the intraspecific level. PMID- 15748856 TI - Resting metabolic expenditure as a potential source of variation in growth rates of the sagebrush lizard. AB - Along an elevational gradient on SW Utah, sagebrush lizards (Sceloporus graciosus) exhibit an unexpected pattern of growth. Lizards from a high elevation population grow faster than lizards from two populations at lower elevations despite shorter daily and seasonal activity. Results from a common environment study of growth suggest that the differences in growth are not due to adaptation to local environmental conditions. In this study, I test the hypothesis that higher growth rates in lizards from high elevation may be attributable to reduced resting metabolic expenditure compared to that of lizards from populations at two lower elevations. Resting metabolic rates were measured for individuals from each of the study populations across different times of day and over a broad range of temperatures. Under the same laboratory conditions, field-caught lizards from the high elevation population exhibited lower metabolic rates when compared to lizards from lower elevations. Daily resting metabolic expenditures were calculated using the observed metabolic rates coupled with estimates of daily activity. Daily resting metabolic expenditure was 50% greater for individuals from the two lower elevation populations, which could result in 12.5% more energy that could be potentially allocated to growth for lizards from high elevation. Such energetic savings may be able to explain differences in the patterns of growth observed in nature. PMID- 15748857 TI - Temperature sensitivity of calcium binding for parvalbumins from Antarctic and temperate zone teleost fishes. AB - Parvalbumin (PV) is a soluble calcium-binding protein that is especially abundant in fast-twitch muscles of fish and other lower vertebrates. Despite its prevalence in ectothermic taxa, few data address the effects of temperature on PV binding function. In this study, calcium dissociation constants (KD) were measured as a function of temperature (0-25 degrees C) for PV from two Antarctic (Gobionotothen gibberifrons and Chaenocephalus aceratus) and two temperate zone fish species (Cyprinus carpio and Micropterus salmoides). Measurements by fluorometric competitive binding assay show that KD values for PVs from the Antarctic species were significantly higher at all assay temperatures and were less sensitive to temperature relative to carp and bass. However, estimates of KD are fundamentally similar for PVs from the Antarctic and temperate zone species when examined at their native physiological temperature. Variation in pH and ionic strength within a physiologically relevant range had only modest effects on KD. Thermodynamics of calcium binding to PV from G. gibberifrons and C. carpio was measured by isothermal microcalorimetry. When measured at 15 degrees C, the Gibbs free energy change (deltaG) was significantly greater for calcium binding to PV from G. gibberifrons than from carp (-43.4+/-1.5 kJ mol(-1) and -46.6+/-3.0 kJ mol(-1), respectively), and the relative contribution of entropy to deltaG for calcium binding to PV from the Antarctic species was about twice that of carp (deltaS=16.0+/-0.8 J degrees C(-1) mol(-1) for G. gibberifrons; deltaS=7.5+/-0.8 J degrees C(-1) mol(-1) for C. carpio). PMID- 15748858 TI - Effect of photoperiod on cultured granulosa cells of the bank vole, Clethrionomys glareolus. AB - Gonadal function of the bank vole females depends on the photoperiod. This experiment was to show whether photoperiod applied on the whole animal in vivo would affect the function of ovarian cells in vitro. Granulosa cells from large ovarian follicles of bank vole reared in long or short photoperiod were cultured as monolayers in control or luteinizing hormone supplemented media. Formation of cell colonies, activity of delta5, 3beta-hydroxy steroid dehydrogenase and progesterone secretion were investigated. First colonies of long day cells were formed already on day 1. On day 2 they enlarged and became abundant. Short day cells formed colonies only on day 2. Colonies of similar size to 2 day colonies of long day cells appeared only on day 6. There were also differences in steroid dehydrogenase activity and in progesterone secretion between long and short day control and hormone treated cultures. We conclude that photoperiod applied in vivo affects ovarian cell function in vitro. PMID- 15748859 TI - Adaptations to fasting in the American mink (Mustela vison): carbohydrate and lipid metabolism. AB - The aim of this study was to investigate whether the actively wintering American mink Mustela vison is strictly dependent on continuous food availability or if it has evolved physiological adaptations to tolerate nutritional scarcity. Fifty farm-bred male minks were divided into a fed control group and four experimental groups fasted for 2, 3, 5 or 7 days. The rate of weight loss was several-fold higher (1.5-3.2% day(-1)) in the mink than recorded previously in larger carnivores utilizing passive wintering strategies. The minks remained normoglycaemic, although their liver glycogen stores and glucose-6-phosphatase activities decreased during fasting. Adipose tissue constituted approximately 36% of their body mass after 7 days of food deprivation. Intra-abdominal fat, especially retroperitoneal but also mesenteric adipose tissue, were the most important fat depots to be hydrolyzed, but the ability of the mink to utilize its body lipids during fasting may be limited. The increased liver size, hepatic triacylglycerol accumulation and increases in the activities of plasma aminotransferases indicated liver dysfunction. Food deprivation also affected the red blood cell indices, and the blood monocyte and lymphocyte counts decreased suggesting immunosuppression during fasting. The results of the present study suggest that the mink has not evolved sophisticated adaptations to wintertime fasting. PMID- 15748860 TI - Distribution of proglucagon mRNA and GLP-1 in the brainstem of chicks. AB - Glucagon-like peptide-1 (GLP-1), structurally similar to glucagon, synthesized from the precursor proglucagon, is a well known anorexigenic peptide in the brain of several animal species. However, there are no previous reports concerning GLP 1-containing neurons in the chick brain. The aim of the present study was to investigate the distribution of proglucagon mRNA and GLP-1-immunoreactive (GLI) perikarya in various regions of the chick brain. We detected proglucagon mRNA in the brainstem, and to a lesser extent in the telencephalon. In the brainstem, a study using immunohistochemistry revealed that GLI perikarya were present in the nucleus motorius nervi facialis pars dosalis, nucleus motoris dorsalis nervi vagi and nucleus tractus solitarii. Furthermore, we found that proglucagon mRNA expression in the brainstem decreased after 24 h fasting. The present findings support the idea that endogenous GLP-1 is involved in feeding behavior of chicks. PMID- 15748861 TI - Heat shock protein induction in the freshwater prawn Macrobrachium malcolmsonii: acclimation-influenced variations in the induction temperatures for Hsp70. AB - The intracellular build-up of thermally damaged proteins following exposure to heat stress results in the synthesis of heat shock proteins (Hsps). In the present study, the upper thermal tolerance and expression of heat shock protein 70 (Hsp70) were examined in juveniles of the freshwater prawn Macrobrachium malcolmsonii that had been acclimated at two different temperatures, i.e. 20 degrees C (group A) and 30 degrees C (group B), in the laboratory for 30 days. Upper thermal tolerance was determined by a standard method. For heat-shock experiments, prawns in groups A and B were exposed to various elevated temperatures for 3 h each, followed by 1 h recovery at the acclimation temperature. Endogenous levels of Hsp70 were determined in the gill, heart, hepatopancreas and skeletal muscle tissues by Western blotting analysis of one dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE). The critical thermal maximum (CT max) for prawns in groups A and B was 37.7+/-0.27 degrees C and 41.41+/-0.16 degrees C, respectively. In general, Western blotting analysis for Hsp70 revealed one band at the 70 kDa region, containing both constitutive (Hsc70) and inducible (Hsp70) isoforms, in the gill and heart tissues; these were not detected in the hepatopancreas and skeletal muscle tissues. The onset temperature for Hsp70 induction in both gill and heart tissues was 30 degrees C for prawns in group A and 34 degrees C for those in group B. The optimum induction temperatures (at which Hsp70 induction was maximum) were found to be 34 degrees C and 32 degrees C, respectively, in the gill and heart tissues of group A prawns, and 38 degrees C and 36 degrees C, respectively, for group B prawns. These results suggest that the temperature at which acclimation occurs influences both upper thermal tolerance and Hsp70 induction in M. malcolmsonii. PMID- 15748862 TI - Role of adiposity hormones in the mouse during fasting and winter acclimatization. AB - The influence of fasting and winter-acclimatization (cold and short-day acclimatization) on mouse plasma leptin, ghrelin, growth hormone (GH) and melatonin concentrations was determined from blood samples taken at mid-day and midnight. A 16-h fast decreased the plasma leptin but almost doubled the plasma ghrelin concentrations which contribute to energy saving, appetite stimulation and, in the case of leptin, to inhibition of reproduction. Winter-acclimatization did not affect plasma ghrelin concentrations, whereas leptin decreased to the same level as in fasting. The low leptin concentrations possibly enable an increased caloric intake for the purpose of thermogenesis. Fasting and winter acclimatization seemed to abolish the diurnal leptin rhythm, but had no effect on that of ghrelin. Plasma melatonin concentration correlated negatively with ghrelin, suggesting a possible role for melatonin in the regulation of ghrelin concentration. SNS-activity and insulin appear to be the main regulators of leptin plasma concentration in the mouse, rather than melatonin as in some seasonal mammals. Interestingly, endogenous ghrelin did not stimulate GH secretion, which is a well-documented reaction to exogenous ghrelin injections. PMID- 15748863 TI - Exhaustive exercise and the cellular stress response in rainbow trout, Oncorhynchus mykiss. AB - The goal of this study was to examine the cellular response to exhaustive exercise in male and female rainbow trout to determine if HSPs are involved in the early stages of the recovery process. Levels of HSPs and key metabolic parameters were measured in white muscle, heart plasma, and blood plasma throughout 6 h of recovery from exhaustive burst exercise. Plasma creatine kinase (CK) was also quantified as an indicator of exercise-induced tissue damage. The observed trends in ATP and lactate were consistent with established patterns of exhaustion and the beginnings of metabolic recovery. However, no upregulation of hsp70, hsp30, or hsp90 was evident in heart or muscle tissue of males or females, and plasma CK measurements suggest that tissue damage was minimal. Our results indicate that hsp70, hsp30, and hsp90 are not part of the early recovery process from burst exercise in fish, perhaps due to the maintenance of core temperatures as well as a lack of exercise-induced tissue damage. PMID- 15748864 TI - Effects of hypoxia on the venous circulation in rainbow trout (Oncorhynchus mykiss). AB - Hypoxia in fish is generally associated with bradycardia while cardiac output (Q) remains unaltered or slightly increased due to a compensatory increase in stroke volume (SV). Rainbow trout (Oncorhynchus mykiss) were subjected to severe (P(W)O2=7.3+/-0.2 kPa) or mild (P(W)O2=11.5+/-0.2 kPa) hypoxia. Central venous pressure (P(ven)), dorsal aortic pressure (P(da)), heart rate (f(H)) and Q, were recorded in vivo. Both levels of hypoxia triggered a significant increase in P(ven). Severe hypoxia was associated with bradycardia and unaltered Q, whereas mild hypoxia was associated with a small but significant increase in Q and no bradycardia. These findings indicate that an increase in P(ven) promotes an increase in SV during hypoxia. Since mild hypoxia increased P(ven), Q and SV without bradycardia or reduced systemic resistance (R(sys)), we hypothesize that an active increase in venous tone serving to mobilize blood to the central venous compartment in order to increase cardiac preload and consequently SV, is an important cardiovascular trait associated with hypoxia. Pharmacological pre treatment with prazosin (1 mg kg(-1)) did not conclusively reveal the underlying mechanisms to the observed changes in P(ven). This study discusses the influence of venous pooling, reduced R(sys) and altered venous tone on changes in P(ven) observed during hypoxia. PMID- 15748866 TI - Adiponectin and peptide YY in the fasting blue fox (Alopex lagopus). AB - Adiponectin (Acrp30) and peptide YY (PYY) are weight-regulatory hormones participating in the control of energy homeostasis. This study investigated the effects of long-term wintertime fasting on plasma Acrp30 and PYY levels in the carnivorous blue fox, a farm-bred variant of the arctic fox (Alopex lagopus). Plasma Acrp30 and PYY concentrations were determined with radioimmunoassays during a 22-day period of fasting, which led to a 20.3% reduction in body mass of the animals (n=32). Sixteen fed blue foxes served as the control group. Acrp30 and PYY were present in blue fox plasma at similar or lower levels as reported previously for other mammals. Fasting had no acute effects on Acrp30 or PYY concentrations of the blue foxes. However, the Acrp30 levels of the fasted blue foxes were 24%-48% higher than in the fed animals between days 8-22 of fasting. Fasted blue foxes also had 6.2-fold higher plasma PYY concentrations after 15 days of fasting. Acrp30 and PYY seem to play roles in the body weight-regulation of the blue fox during long-term fasting, but their specific functions and physiological significance remain to be determined. PMID- 15748865 TI - Haemolymph glucose concentrations of juvenile rock lobsters, Jasus edwardsii, feeding on different carbohydrate diets. AB - Postprandial changes in haemolymph glucose concentration ([Glc]H) were measured in 4-day-fasted juvenile intermoult spiny lobsters, Jasus edwardsii, provided with meals composed of glycogen, maltose, sucrose, glucose, or fructose in a gelatine base, or with gels of the algal glycans agar, alginate and carrageenan. Baseline [Glc]H was 0.61+/-0.02 mmol L(-1). After consumption of glycogen, maltose or sucrose, [Glc]H approximately doubled, peaked after 3 h and returned to baseline between 12 and 24 h. Glucose and fructose meals were followed by periods of sustained hyperglycaemia lasting more than 24 h (peaking at approximately 2.5 times baseline at 6 and 3 h respectively). Suggested explanations for augmented hyperglycaemic responses to glucose and fructose are: 1) these monosaccharides by-passed contact digestion and absorption in the R cells of the digestive gland, directing them away from storage and toward transepithelial scavenging routes; or 2) glucose and fructose directly elicited release of crustacean hyperglycaemic hormone via a chemosensory reflex. Agar and alginate induced significant postprandial glycaemic responses, consistent with reports of carbohydrases in this species and indicating their potential for inclusion in artificial diets as both binders and energy sources. Carrageenan, a highly sulphated galactan, did not produce a glycaemic response. The measurement of glycaemic responses is a quick method of obtaining nutritional information on carbohydrates considered for inclusion in formulated diets prior to lengthy growth trials. PMID- 15748867 TI - The differences between high and low-dose administration of VEGF to dopaminergic neurons of in vitro and in vivo Parkinson's disease model. AB - Vascular endothelial growth factor (VEGF) has previously been shown to display neuroprotective effects on dopaminergic (DA) neurons. In this study, we investigated whether the effects of VEGF were dose-dependent or not. First, VEGF was shown to be neuroprotective on 6-hydroxydopamine (6-OHDA)-treated murine DA neurons in vitro, although the 1 ng/ml of VEGF displayed more neuroprotective effects than 100 ng/ml. Furthermore, using 2 sizes of capsules (small/large) with different secreting quantities, 6-OHDA-treated rats receiving the small capsule filled with VEGF-secreting cells (BHK-VEGF) into the striatum showed a significant decrease in amphetamine-induced rotational behavior in number and a significant preservation of TH-positive fibers compared to those receiving the large BHK-VEGF capsule as well as those receiving BHK-Control capsule. Rats receiving the large BHK-VEGF capsule showed much more glial proliferation, angiogenesis, and brain edema around the capsule than those with the small one. High-dose administration of VEGF might cause poor circulation related to brain edema, although low-dose administration of VEGF displays neuroprotective effects on DA neurons. Our results demonstrate the importance of administration dose of VEGF, suggesting that low-dose administration of VEGF might be desirable for Parkinson's disease. PMID- 15748868 TI - Regional expression of constitutive and inducible transcription factors following transient focal ischemia in the neonatal rat: influence of hypothermia. AB - Ischemia is a potent modulator of gene expression. Differential expression of transcription factors after focal ischemia may reflect the potential for neuronal recovery in peri-ischemic regions. Previously, we demonstrated that hypothermia reduces the volume of damage in a model of neonatal focal ischemia. In the present study, immunocytochemistry was used to assess the temporal and spatial profiles of the transcription factors Fos and pCREB under normal and hypothermic conditions in this neonatal model of focal ischemia. At 7 days of age, rat pups underwent a permanent middle cerebral artery occlusion (MCAo) coupled with a temporary 1-h occlusion of the common carotid artery (CCAo). They were maintained at 37 degrees C throughout ischemia and reperfusion (Normothermic), or given 1 h of hypothermic conditions (28 degrees C) either during the occlusion (Intraischemic Hypothermia) or during the second hour of reperfusion (postischemic hypothermia). In normothermic pups, Fos immunoreactivity peaked at early time points (4-8 h post-ischemia) in a narrow band in peri-ischemic regions. By later stages of reperfusion (12-24 h), there was a more widespread induction in peri-ischemic regions including the ipsilateral cortex. In contrast with Fos, the constitutive transcription factor pCREB was reduced in core regions at all time points examined. Both the c-fos induction in peri-ischemic regions and the reduction of pCREB in the core were attenuated by intraischemic hypothermia. Postischemic hypothermia altered the distribution of Fos immunoreactivity without significantly changing the number of Fos- and pCREB immunoreactive cells compared to normothermic rats. Both intra- and postischemic hypothermia reduced the number of caspase-immunoreactive cells. Thus, focal ischemia in the P7 rat produces different distributions of Fos and pCREB than what has been observed in adult rats subjected to focal ischemia, and expression of these transcription factors can be altered by hypothermia. PMID- 15748869 TI - Neuronal and astroglial alterations in the hippocampus of a mouse model for type 1 diabetes. AB - The influence of diabetes mellitus on brain pathology is increasingly recognized. Previous contributions of our laboratory demonstrated in models of type 1 diabetes (nonobese diabetic and streptozotocin (STZ)-treated mice), a marked astrogliosis and neurogenesis deficit in hippocampus and increased expression of hypothalamic neuropeptides. In the present investigation, we further analyzed alterations of astroglia and neurons in the hippocampus of mice 1 month after STZ induced diabetes. Results showed that these STZ-diabetic mice presented: (a) increased number of astrocytes positive for apolipoprotein-E (Apo-E), a marker of ongoing neuronal dysfunction; (b) abnormal expression of early gene products associated with neuronal activation, including a high number of Jun + neurons in CA1 and CA3 layers and dentate gyrus, and of Fos-expressing neurons in CA3 layer; (c) augmented activity of NADPH-diaphorase, linked to oxidative stress, in CA3 region. These data support the concept that uncontrolled diabetes leads to hippocampal pathology, which adjoin to changes in other brain structures such as hypothalamus and cerebral cortex. PMID- 15748870 TI - Vasopressinergic hypothalamic neurons are recruited during the audiogenic seizure of WARs. AB - The Wistar Audiogenic Rat (WAR) is a genetic model of reflex epilepsy with seizures induced by high-intensity sound stimulation (120 dB SPL). In spite of the known neural substrates involved in WAR seizure phenotype, neuroendocrine hypothalamic neurons were never investigated. In this work, AVP immunohistochemistry in the hypothalamus and radioimmunoassay (RIA) in plasma and in hypothalamic and hypophysial tissues were performed on both controls and WARs in order to evaluate the dynamics of AVP release due to seizure induction. Susceptible animals (WARs) displayed at least tonic-clonic convulsions followed by clonic spasms, while resistant Wistar rats (R) had no convulsive behavior. Animals were sacrificed at 3 instances: basal condition (without stimulus) and at 3 and 10 min after sound stimulation. For the immunohistochemistry AVP study, brains were harvested and processed by the avidin-biotin-peroxidase detection method. Optic densitometry was used for quantifying AVP labeling in supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. SON presented higher densitometry levels (%D--relative to background) for both WARs and R when compared to PVN. Nevertheless, both nuclei presented a marked decrease, referenced to basal levels, in %D for WARs at 3 min (approximately 35%) against a discrete change for R (approximately 90%). RIA results were significantly higher in the hypophysis of WARs when compared to R rats, at 3 min. Also, at 3 min, plasma AVP in WARs (89.32 +/- 24.81 pg/mL) were higher than in R (12.01 +/- 2.39 pg/mL). We conclude, based on the AVP releasing profiles, that vasopressinergic hypothalamic neurons are recruited during the audiogenic seizure of WARs. PMID- 15748871 TI - Neural precursor cells division and migration in neonatal rat brain after ischemic/hypoxic injury. AB - Ischemia/hypoxia (I/H) causes severe perinatal brain disorders such as cerebral palsy. The neonatal brain possesses much plasticity, and to enhance new cell production would be an innovative means of therapy for such disorders. In order to elucidate the dynamic changes of neural progenitor cells in the neonatal brain after ischemia, we investigated new cells production in the subventricular zone and subsequent migration of these cells to the injured area. Newly produced cells were confirmed by incorporation of bromodeoxyuridine (BrdU), and attempt for differentiation was investigated by immunohistochemistry for molecular markers of each cellular lineage. In the sham-control brain, there were many BrdU-labeled cells which gradually decreased as the animal becomes older. Many of these cells were oligodendroglial progenitor or microglial cells. Although there were only few neuronal cells labeled for BrdU in the sham-control, they dramatically increased after I/H. They were located at just beneath the subventricular zone where the progenitor cells reside and to the injured area, indicating that newly produced cells migrated to the infarct region and differentiated into neuronal precursor cells in order to compensate the lost neural cells. We found that BrdU labeled astroglial, oligodendroglial progenitor, and microglial cells were also increased after I/H, suggesting that they also play active roles in recovery. Progenitor cells may have potential for treating perinatal brain disorders. PMID- 15748872 TI - Methanesulfonyl fluoride, an acetylcholinesterase inhibitor, attenuates simple learning and memory deficits in ischemic rats. AB - Methanesulfonyl fluoride (MSF), a highly selective CNS inhibitor of acetylcholinesterase, has been recently demonstrated to promote improvement in cognitive performance in patients with senile dementia of Alzheimer type. Because a similar cognitive impairment may accompany stroke, we investigated in the present study whether treatment with MSF could produce beneficial effects in adult rats subjected to an experimental stroke model. Sprague-Dawley rats received transient 60 min intraluminal occlusion of the right middle cerebral artery (MCAo) and were given i.p. injections of either MSF (1 mg/kg at 24 and 48 h post-MCAo and 0.3 mg/kg thereafter every other day) or the vehicle, peanut oil, for 4 weeks. Behavioral tests and biochemical assays were performed at 28 days post-surgery. MSF treatment produced about 90% inhibition of acetylcholinesterase in the brain. Ischemic animals that received the vehicle displayed significant elevated body swing biased activity (84.8 +/- 10%) and significantly prolonged acquisition (398 +/- 62 s) and shortened retention (79 +/- 26 s) of the passive avoidance task. Interestingly, while the ischemic animals that received the MSF exhibited elevated body swing biased activity (87.7 +/- 8%), they performed significantly better in the passive avoidance task (255 +/- 36 s and 145 +/- 18 s in acquisition and retention) than the vehicle-treated animals. Moreover, whereas brains from both groups of animals revealed similar extent and degree of cerebral infarction, the MSF-treated ischemic animals showed more intense immunoreactivity, as well as a significantly higher number (10-15% increase) of septal choline acetyltransferase-positive cells than the vehicle-treated ischemic animals. These results show that MSF, possibly by preserving a functional cholinergic system, attenuated stroke-induced deficits in a simple learning and memory task. PMID- 15748873 TI - Progesterone's 5 alpha-reduced metabolite, 3 alpha,5 alpha-THP, mediates lateral displacement of hamsters. AB - 5 alpha-Pregnan-3 alpha-ol-20-one (3 alpha,5 alpha-THP), progesterone (P4)'s 5 alpha-reduced, 3 alpha-hydroxysteroid oxidoreduced product, facilitates lordosis of rodents in part via agonist-like actions at GABA(A)/benzodiazepine receptor complexes in the ventral tegmental area (VTA). Whether 3 alpha,5 alpha-THP influences another reproductively-relevant behavior, lateral displacement, of hamsters was investigated. Lateral displacement is the movement that female hamsters make with their perineum towards male-like tactile stimulation. This behavior facilitates, and is essential for, successful mating. Hamsters in behavioral estrus had greater lateral displacement responses when endogenous progestin levels were elevated compared to when progestin levels were lower. Administration of P4, a prohormone for 3 alpha,5 alpha-THP, dose-dependently (500 > 200 > 100, 50, or 0 microg) enhanced lateral displacement of ovariectomized hamsters that had been primed with SC estradiol benzoate (5 or 10 microg). Inhibiting P4's metabolism to 3 alpha,5 alpha-THP by co-administering finasteride, a 5 alpha-reductase inhibitor, or indomethacin, a 3 alpha hydroxysteroid oxidoreductase inhibitor, either systemically or to the VTA, significantly decreased lateral displacement and midbrain progestin levels of naturally receptive or hormone-primed hamsters compared to controls. These data suggest that lateral displacement is progestin-sensitive and requires the formation of 3 alpha,5 alpha-THP in the midbrain VTA. PMID- 15748874 TI - Gustatory stimulation influences the processing of intranasal stimuli. AB - OBJECTIVES: Taste and smell interact. The aim of this study was to examine this interaction using gustatory and olfactory stimuli applied at the same time, which exhibited perceptual compatibility and incompatibility. METHODS: Thirty-two, young, healthy normosmic subjects (16 men, 16 women) took part in two randomized sessions. Event-related potentials (ERP) were recorded in response to vanillin, or gaseous CO2. These two conditions were combined with three "taste conditions" including sweet taste, sour taste, and the intraoral presentation of an empty taste dispenser. RESULTS: Vanillin responses were largest for the "sweet" condition, while they were smaller for the "sour condition". In contrast, responses to CO2 were largest under the "sour" condition, and smallest under the "sweet" condition. Moreover, during the "sweet" condition the latencies of P1 and N1 were shorter than in the "sour" condition, which was the other way around for CO2. CONCLUSIONS: Results of the present investigation suggested that (1) the early processing of intranasal chemosensory stimuli is modulated through concomitant gustatory stimulation, and that (2) this modulation may depend, at least in part, on the contextual compatibility between intranasal and intraoral stimuli. PMID- 15748875 TI - Spinal nerve ligation increases alpha2-adrenergic receptor G-protein coupling in the spinal cord. AB - Intrathecal and epidural administration of the alpha2-adrenergic receptor agonist clonidine in humans results in analgesia to both acute nociceptive and chronic neuropathic pain. The potency of clonidine increases with hypersensitivity to mechanical stimuli after nerve injury, although the reasons for this change are unknown. In the present study, we tested the hypothesis that peripheral nerve injury alters either spinal alpha2-adrenergic receptor-mediated G-protein activity or alpha2-adrenergic receptor number. Rats were randomized to left spinal nerve ligation (SNL) or sham surgery. Tactile hypersensitivity in the hindpaw was confirmed and lumbar spinal cords were removed for binding assays. To examine agonist-induced G-protein coupling, [35S]GTP gamma S binding experiments were performed in spinal cord membranes and sections using norepinephrine as an alpha2-adrenergic agonist. SNL was associated with an increase in maximal efficacy, but not potency, of norepinephrine-stimulated [35S]GTP gamma S binding in dorsal horn. SNL had no effect on basal [35S]GTP gamma S binding or on muscarinic cholinergic-stimulated [35S]GTP gamma S binding. [35S]GTP gamma S autoradiography showed that this increase in alpha2-adrenergic-activated G proteins occurred both ipsilateral and contralateral to SNL surgery. SNL did not alter total alpha2-adrenergic receptor number or affinity to [3H]-rauwolscine binding, and displacement studies with the alpha2A-adrenergic antagonist BRL44408 revealed that most of the binding was associated with the alpha2A-adrenergic subtype. These data suggest that the increased potency of clonidine in neuropathic pain could reflect increased efficiency of G-protein coupling from spinal alpha2-adrenergic receptors. PMID- 15748876 TI - Failure of FK506 (tacrolimus) to alleviate apomorphine-induced circling in rat Parkinson model in spite of some cytoprotective effects in SH-SY5Y dopaminergic cells. AB - The mechanism of action of the neurotoxin 6-hydroxydopamine (6-OHDA) is thought to involve the generation of free radicals and subsequent apoptotic processes. We have demonstrated in vitro that the neuroimmunophilin, FK506 (10-100 nM), dose dependently and significantly restored the ROS production to the control level, increased the Bcl-2 protein level, partly inhibited the cytochrome C release from mitochondria and reduced the caspase-3 activation in SH-SY5Y cells. On the other hand, there was no significant restoration of the ATP level by FK506 and the toxin activated proteins, p53 and Bax, were not normalized by FK506. In support of these latter results, daily administration of FK506 for 7 days to rats (0.5, 1 and 3 mg/kg i.p.) did not significantly prevent the apomorphine-induced contralateral circling, measured 2 weeks after unilateral nigral lesioning. Moreover, FK506 pretreatment did not significantly lower the toxin elevated lipid peroxidation levels, indicating that oxidative stress was present even after the FK506 treatment in the lesioned striatum. Taken together, our results with FK506 are inconsistent. We confirm the antioxidant nature of FK506, that is, it blocks ROS production in SH-SY5Y cells. However, there were no significant protective effects in any apoptotic analyses in SH-SY5Y cells and in animal studies, a 7-day FK506 pre-treatment was not able to reverse the toxic effect of 6-OHDA in a rat model of Parkinson's disease. PMID- 15748877 TI - The effect of glutamate receptor blockers on glutamate release following spinal cord injury. Lack of evidence for an ongoing feedback cascade of damage --> glutamate release --> damage --> glutamate release --> etc. AB - It is widely hypothesized that excitotoxicity of released glutamate following a CNS insult is propagated by the cyclic cascade: glutamate release --> damage --> glutamate release --> further damage --> etc. We tested this hypothesis by determining the effects of attempting to interrupt the loop by administering glutamate receptor antagonists and Na(+)-channel blockers on glutamate release following spinal cord injury (SCI). The effects of administering the NMDA receptor blockers MK-801 and memantine, the AMPA/kainate receptor blockers NBQX and GYKI 52466, the AMPA receptor desensitization blocker cyclothiazide and the sodium channel blockers riluzole, mexiletine and QX-314 on post-SCI were determined. Agents were administered into the site of injury by direct injection, by microdialysis or systemically. None of these agents had an appreciable effect on glutamate release following SCI. Thus, it is unlikely that the above cascade produces significant secondary glutamate release and ongoing damage following SCI, although such cascades may worsen other CNS insults. We attribute our results to overwhelming effects of much greater release by direct mechanical damage and reversal of transport following SCI. PMID- 15748878 TI - Magnesium deficiency impairs fear conditioning in mice. AB - Magnesium (Mg2+) is one of the most abundant cations found in the body. In the central nervous system, Mg2+ plays an important role in the function of N-methyl D-aspartate (NMDA)-type glutamate receptors, which are centrally involved in memory processing. Despite the relatively large concentration of Mg2+ in the CNS, little is known about the behavioral consequences of Mg2+ deficiency. The purpose of this study was to address this issue by assessing fear conditioning and related behaviors in mice maintained on normal or Mg(2+)-deficient diets. Young adult male C57Bl/6J mice were placed on a control or Mg(2+)-deficient diet, and testing was conducted between 10 and 21 days later. Magnesium-deficient mice exhibited impairments in contextual and cued fear conditioning. These impairments could not be attributed to changes in locomotor activity, exploration, or pain sensitivity. Furthermore, Mg(2+)-deficient mice were more sensitive to the convulsant effects of a peripheral injection of NMDA (100 mg/kg, IP). The results suggest that magnesium deficiency can lead to specific impairments in emotional memory. Such impairments may be related to hypersensitivity of NMDA-type glutamate receptors in Mg(2+)-deficient mice. PMID- 15748879 TI - Peptide 19 in the rat vagal and glossopharyngeal sensory ganglia. AB - Peptide 19 (PEP 19) is a 7.6-kDa polypeptide which binds to calmodulin and inhibits calcium-calmodulin signaling. In this study, PEP 19-immunoreactivity (PEP 19-IR) was examined in the rat vagal and glossopharyngeal sensory ganglia. Twenty-nine percent, 59%, and 41% of sensory neurons contained PEP 19-IR in the jugular, petrosal, and nodose ganglia, respectively. These neurons were of various sizes (jugular, mean +/- SD = 635.8 +/- 392.6 microm2, range = 105.9 1695.9 microm2; petrosal, mean +/- SD = 370.9 +/- 228.5 microm2, range = 57.7 1662.7 microm2; nodose, mean +/- SD = 380.5 +/- 157 microm2, range = 87.5-950.4 microm2) and scattered throughout these ganglia. Double immunofluorescence method revealed that PEP 19-IR neurons which had parvalbumin-IR were rare in the ganglia (jugular, 4%; petrosal, 10%; nodose, 8%). PEP 19-IR neurons which contained calbindin D-28k were abundant in the petrosal (20%) and nodose (22%) ganglia but not in the jugular ganglion (8%). Retrograde tracing method indicated that many PEP 19-IR neurons projected to the circumvallate papilla and soft palate. In the soft palate, taste buds were innervated by PEP 19-IR nerve fibers. The present study suggests that PEP 19-IR neurons include chemoreceptors in the vagal and glossopharyngeal sensory ganglia. PMID- 15748881 TI - Effects of CGS-12066A on medial vestibular nuclear neurons. AB - This study aims to explore the effects of a selective 5-HT1B receptor agonist, CGS-12066A, on the neuronal excitability of the rat medial vestibular nuclear neurons. The spontaneous firing rate was decreased, and the membrane potential was hyperpolarized by CGS-12066A. The whole potassium currents were inhibited by CGS-12066A. After the calcium-dependent potassium, currents were blocked, however, CGS-12066A did not inhibit the potassium currents, suggesting that the 5 HT action site is calcium-dependent potassium currents. PMID- 15748880 TI - Involvement of cyclic GMP-dependent protein kinase in nitrous oxide-induced anxiolytic-like behavior in the mouse light/dark exploration test. AB - The second messenger cyclic GMP (cGMP) plays a role in the anxiolytic-like behavioral response of mice to nitrous oxide (N2O). This study was conducted to determine whether this behavioral effect of N2O is affected by inhibition of cGMP dependent protein kinase (PKG). N2O-induced behavior in the light/dark exploration test was significantly attenuated by the PKG inhibitors H-8 and Rp-8 pCPT-cGMPS but not Rp-8-pCPT-cAMPS, an inhibitor of cAMP-dependent protein kinase. These findings implicate PKG in the mediation or modulation of the anxiolytic-like behavioral response to N2O. PMID- 15748882 TI - Overexpression of Rab22a hampers the transport between endosomes and the Golgi apparatus. AB - The transport and sorting of soluble and membrane-associated macromolecules arriving at endosomal compartments require a complex set of Rab proteins. Rab22a has been localized to the endocytic compartment; however, very little is known about the function of Rab22a and inconsistent results have been reported in studies performed in different cell lines. To characterize the function of Rab22a in endocytic transport, the wild-type protein (Rab22a WT), a hydrolysis-deficient mutant (Rab22a Q64L), and a mutant with reduced affinity for GTP (Rab22a S19N) were expressed in CHO cells. None of the three Rab22a constructs affected the transport of rhodamine-dextran to lysosomes, the digestion of internalized proteins, or the lysosomal localization of cathepsin D. In contrast with the mild effect of Rab22a on the endosome-lysosome route, cells expressing Rab22a WT and Rab22a Q64L presented a strong delay in the retrograde transport of cholera toxin from endosomes to the Golgi apparatus. Moreover, these cells accumulated the cation independent mannose 6-phosphate receptor in endosomes. These observations indicate that Rab22a can affect the trafficking from endosomes to the Golgi apparatus probably by promoting fusion among endosomes and impairing the proper segregation of membrane domains required for targeting to the trans-Golgi network (TGN). PMID- 15748883 TI - Geranylgeranylated, but not farnesylated, RhoB suppresses Ras transformation of NIH-3T3 cells. AB - RhoB is a low molecular weight GTPase that is both farnesylated (RhoB-F) and geranylgeranylated (RhoB-GG) in cells. Based on data from rodent cell models, it has been suggested that RhoB displays differential effects on cell transformation, according to the nature of its prenylation. To test directly this hypothesis, we generated GTPase-deficient RhoB mutants that are exclusively either farnesylated or geranylgeranylated. We show that in Ras-transformed murine NIH-3T3 cells, RhoB-F enhances, whereas RhoB-GG and RhoB (F/GG) suppresses anchorage-dependent and -independent cell growth as well as tumor growth in nude mice. We then demonstrate that Ras constitutive activation of the tumor survival pathways Akt and NF-kappa B are blocked by RhoB-GG, but not by RhoB-F, providing further support for the opposing role of RhoB-F and RhoB-GG in Ras malignant transformation in NIH-3T3 cells. In addition, both RhoB (F/GG) and RhoB-GG induce apoptosis in Ras-transformed NIH-3T3 cells whereas RhoB-F has no effect. Our data demonstrate that RhoB-F and RhoB-GG which differ only by a 5-carbon isoprene behave differently in rodent cells highlighting the important role of prenyl groups in protein function and emphasize the potency of RhoB to regulate negatively the oncogenic signal. PMID- 15748884 TI - Localization of UDP-GlcNAc 2-epimerase/ManAc kinase (GNE) in the Golgi complex and the nucleus of mammalian cells. AB - The bifunctional enzyme UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE) is essential for early embryonic development and catalyzes the rate limiting step in sialic acid biosynthesis. Although epimerase and kinase activities have been attributed to GNE, little is known about the regulation, differential expression, and subcellular localization of GNE in vivo. Mutations in GNE cause a rare inherited muscle disorder in humans called hereditary inclusion body myopathy (HIBM). However, the role of GNE in HIBM pathogenesis has not been defined yet. Here, we show that the GNE protein is expressed in various mammalian cells and tissues with highest levels found in cancer cells and liver. In human skeletal muscle, GNE protein is developmentally regulated: high levels are found in immature myoblasts but low levels in mature skeletal muscle. The GNE protein colocalizes with resident proteins of the Golgi compartment in a variety of human cells including muscle. Drug-induced disruption of the Golgi and subsequent recovery reveals co-distribution of GNE along with Golgi-targeted proteins. This subcellular localization of GNE is in good agreement with its established role as the key enzyme of sialic acid biosynthesis, since the sialylation of glycoconjugates takes place in the Golgi complex. Surprisingly, GNE is also detected in the nucleus. Upon nocodazole treatment, GNE redistributes to the cytoplasm suggesting that GNE may act as a nucleocytoplasmic shuttling protein. A regulatory role for GNE shifting between the nuclear and the Golgi compartment is proposed. Further insight into GNE regulation may promote the understanding of HIBM pathogenesis. PMID- 15748885 TI - Redox control of angiogenic factors and CD31-positive vessel-like structures in mouse embryonic stem cells after direct current electrical field stimulation. AB - The molecular mechanisms driving angiogenesis in tissues derived from embryonic stem (ES) cells are currently unknown. Herein we investigated the effects of direct current (DC) electrical field treatment on endothelial cell differentiation and angiogenesis of mouse ES cells. Treatment of ES cell-derived embryoid bodies with field strengths ranging from 250 V/m to 750 V/m, applied for 60 s, dose-dependently increased the capillary area staining positive for the endothelial-specific marker platelet endothelial cell adhesion molecule-1 (PECAM 1), indicating stimulation of endothelial cell differentiation and angiogenesis. Consequently, increased expression of hypoxia-inducible factor-1alpha (HIF 1alpha) and vascular endothelial growth factor (VEGF) within 24 h was observed. Electric field treatment raised reactive oxygen species (ROS) generation for at least 48 h, which was blunted by NADPH-oxidase inhibitors diphenylen iodonium chloride (DPI) as well as 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), and increased the expression of NADPH-oxidase subunits p22-phox, p47-phox, p67-phox, and gp91-phox within 24 h. Electrical field treatment resulted in activation of extracellular regulated kinase 1,2 (ERK1,2), p38, as well as c-Jun NH2-terminal kinase (JNK). Pretreatment with the JNK inhibitor SP600125 resulted in a significant decrease in capillary areas under control conditions as well as under conditions of electrical field treatment, whereas the p38 inhibitor SB203580 was without effects. By contrast, the ERK1,2 antagonist UO126 inhibited electrical field-induced angiogenesis, whereas angiogenesis under control conditions was unimpaired. The increase in capillary areas and VEGF expression as well as activation of JNK and ERK1,2 was significantly inhibited in the presence of the free radical scavenger vitamin E underscoring the role of ROS in electrical field induced angiogenesis of ES cells. PMID- 15748886 TI - The cytoplasmic C-terminus of polycystin-1 increases cell proliferation in kidney epithelial cells through serum-activated and Ca(2+)-dependent pathway(s). AB - Polycystin-1 (PC1) is a large transmembrane protein important in renal differentiation and defective in most cases of autosomal dominant polycystic kidney disease (ADPKD), a common cause of renal failure in adults. Although the genetic basis of ADPKD has been elucidated, molecular and cellular mechanisms responsible for the dysregulation of epithelial cell growth in ADPKD cysts are still not well defined. We approached this issue by investigating the role of the carboxyl cytoplasmic domain of PC1 involved in signal transduction on the control of kidney cell proliferation. Therefore, we generated human HEK293 cells stably expressing the PC1 cytoplasmic tail as a membrane targeted TrkA-PC1 chimeric receptor protein (TrkPC1). We found that TrkPC1 increased cell proliferation through an increase in cytoplasmic Ca2+ levels and activation of PKC alpha, thereby upregulating D1 and D3 cyclin, downregulating p21waf1 and p27kip1 cyclin inhibitors, and thus inducing cell cycle progression from G0/G1 to the S phase. Interestingly, TrkPC1-dependent Ca2+ increase and PKC alpha activation are not constitutive, but require serum factor(s) as parallel component. In agreement with this observation, a significant increase in ERK1/2 phosphorylation was observed. Consistently, inhibitors specifically blocking either PKC alpha or ERK1/2 prevented the TrkPC1-dependent proliferation increase. NGF, the TrkA ligand, blocked this increase. We propose that in kidney epithelial cells the overexpression of PC1 C-terminus upregulates serum-evoked intracellular Ca2+ by counteracting the growth-suppression activity of endogenous PC1 and leading to an increase in cell proliferation. PMID- 15748887 TI - Effects of PGF2alpha on human melanocytes and regulation of the FP receptor by ultraviolet radiation. AB - Prostaglandins are potent lipid hormones that activate multiple signaling pathways resulting in regulation of cellular growth, differentiation, and apoptosis. In the skin, prostaglandins are rapidly released by keratinocytes following ultraviolet radiation and are chronically present in inflammatory skin lesions. We have shown previously that melanocytes, which provide photoprotection to keratinocytes through the production of melanin, express several receptors for prostaglandins, including the PGE2 receptors EP1 and EP3 and the PGF2alpha receptor FP, and that PGF2alpha stimulates melanocyte dendricity. We now show that PGF2alpha stimulates the activity and expression of tyrosinase, the rate limiting enzyme in melanin synthesis. Analysis of FP receptor regulation showed that the FP receptor is regulated by ultraviolet radiation in melanocytes in vitro and in human skin in vivo. We also show that ultraviolet irradiation stimulates production of PGF2alpha by melanocytes. These results show that PGF2alpha binding to the FP receptor activates signals that stimulate a differentiated phenotype (dendricity and pigmentation) in melanocytes. The regulation of the FP receptor and the stimulation of production of PGF2alpha in melanocytes in response to ultraviolet radiation suggest that PGF2alpha could act as an autocrine factor for melanocyte differentiation. PMID- 15748888 TI - Cell responses to FGFR3 signalling: growth, differentiation and apoptosis. AB - FGFR3 is a receptor tyrosine kinase (RTK) of the FGF receptor family, known to have a negative regulatory effect on long bone growth. Fgfr3 knockout mice display longer bones and, accordingly, most germline-activating mutations in man are associated with dwarfism. Somatically, some of the same activating mutations are associated with the human cancers multiple myeloma, cervical carcinoma and carcinoma of the bladder. How signalling through FGFR3 can lead to either chondrocyte apoptosis or cancer cell proliferation is not fully understood. Although FGFR3 can be expressed as two main splice isoforms (IIIb or IIIc), there is no apparent link with specific cell responses, which may rather be associated with the cell type or its differentiation status. Depending on cell type, differential activation of STAT proteins has been observed. STAT1 phosphorylation seems to be involved in inhibition of chondrocyte proliferation while activation of the ERK pathway inhibits chondrocyte differentiation and B-cell proliferation (as in multiple myeloma). The role of FGFR3 in epithelial cancers (bladder and cervix) is not known. Some of the cell specificity may arise via modulation of signalling by crosstalk with other signalling pathways. Recently, inhibition of the ERK pathway in achondroplastic mice has provided hope for an approach to the treatment of dwarfism. Further understanding of the ability of FGFR3 to trigger different responses depending on cell type and cellular context may lead to treatments for both skeletal dysplasias and cancer. PMID- 15748889 TI - ACTH-induced caveolin-1 tyrosine phosphorylation is related to podosome assembly in Y1 adrenal cells. AB - Y1 adrenocortical cells respond to ACTH with a characteristic rounding-up that facilitates cAMP signaling, critical for transport of cholesterol to the mitochondria and increase in steroid secretion. We here demonstrate that caveolin 1 participates in coupling activation of protein kinase A (PKA) to the control of cell shape. ACTH/8-Br-cAMP induced reorganization of caveolin-1-positive structures in correlation with the cellular rounding-up. Concomitant with this change, there was an increase in the phosphorylation of caveolin-1 (Tyr-14) localized at focal adhesions (FA) with reorganization of FA to rounded, ringlike structures. Colocalization with phalloidin showed that phosphocaveolin is present at the edge of actin filaments and that after ACTH stimulation F-actin dots at the cell periphery become surrounded by phosphocaveolin-1. These observations along with electron microscopy studies revealed these structures as podosomes. Podosome assembly was dependent on both PKA and tyrosine kinase activities because their formation was impaired after treatment with specific inhibitors [myristoylated PKI (mPKI) or PP2, respectively] previous to ACTH/8-Br-cAMP stimulation. These results show for the first time that ACTH induces caveolin-1 phosphorylation and podosome assembly in Y1 cells and support the view that the morphological and functional responses to PKA activation in steroidogenic cells are related to cytoskeleton dynamics. PMID- 15748890 TI - The actin cytoskeleton-associated protein zyxin acts as a tumor suppressor in Ewing tumor cells. AB - Changes in cell architecture, essentially linked to profound cytoskeleton rearrangements, are common features accompanying cell transformation. Supporting the involvement of the microfilament network in tumor cell behavior, several actin-binding proteins, including zyxin, a potential regulator of actin polymerization, may play a role in oncogenesis. In this work, we investigate the status of zyxin in Ewing tumors, a family of pediatric malignancies of bone and soft tissues, which are mainly associated with a t(11;22) chromosomal translocation encoding the EWS-FLI1 oncoprotein. We observe that EWS-FLI1 transformed murine fibroblasts, as well as human Ewing tumor-derived SK-N-MC cells, exhibit a complete disruption of their actin cytoskeleton, retaining very few stress fibers, focal adhesions and cell-to-cell contacts. We show that within these cells, zyxin is expressed at very low levels and remains diffusely distributed throughout the cytoplasm, instead of concentrating in actin-rich dynamic structures. We demonstrate that zyxin gene transfer into EWS-FLI1 transformed fibroblasts elicits reconstitution of zyxin-rich focal adhesions and intercellular junctions, dramatic reorganization of the actin cytoskeleton, decreased cell motility, inhibition of anchorage-independent growth and impairment of tumor formation in athymic mice. We observe similar phenotypic changes after zyxin gene transfer in SK-N-MC cells, suggesting that zyxin has tumor suppressor activity in Ewing tumor cells. PMID- 15748891 TI - Dynamics and compartmentation of the nucleolar processing machinery. AB - In active nucleoli, machineries involved in the biogenesis of ribosomal RNAs (rRNAs) are compartmentalized. The late rRNA processing proteins are localized in the granular component (GC). Here we investigate the behavior of these proteins when production of 28S is impaired and when this blockage is reversed. The 5,6 dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) provokes dispersion of rDNA clusters and we demonstrate that DRB induces disconnection of the late rRNA processing proteins from the transcription sites. These processing proteins are still associated in independent masses without detectable 28S rRNA, indicating that compartmentation of the late rRNA processing machinery is not necessarily linked to processing activity. Removing DRB reverses this disconnection and promotes rRNA processing. Nucleolar reformation occurs in two successive steps, dynamic recruitment to transcription sites of the processing proteins, followed by rDNA compaction. We demonstrate that both steps are sensitive to temperature, suggesting an energy-dependent process. Traffic of processing proteins analyzed by fluorescence recovery after photobleaching is similar in masses disconnected from transcription sites and in the granular component of the active nucleolus. This suggests that protein dynamics and interactions, and not only their processing activity, determine compartmentation of the nucleolar machineries. PMID- 15748892 TI - Keratin-containing inclusions affect cell morphology and distribution of cytosolic cellular components. AB - Many neurodegenerative diseases are characterized by the presence of protein aggregates bundled with intermediate filaments (IFs) and similar structures, known as Mallory bodies (MBs), are observed in various liver diseases. IFs are anchored at desmosomes and hemidesmosomes, however, interactions with other intercellular junctions have not been determined. We investigated the effect of IF inclusions on junction-associated and cytosolic proteins in various cultured cells. We performed gene transfection of the green fluorescent protein (GFP) tagged cytokeratin (CK) 18 mutant arg89cys (GFP-CK18 R89C) in cultured cells and observed CK aggregations as well as loss of IF networks. Among various junction associated proteins, zonula occludens-1 and beta-catenin were colocalized with CK aggregates on immunofluorescent analyses. Similar results were obtained on immunostaining for cytosolic proteins, 14-3-3 zeta protein, glucose-6-phosphate dehydrogenase and DsRed. E-cadherin, a basolateral membrane protein in polarized epithelia, was present on both the apical and basolateral domains in GFP-CK18 R89C-transfected cells. Furthermore, cells containing CK aggregates were significantly larger than GFP-tagged wild type CK18 (GFP-WT CK18)-transfected or non-transfected cells (P < 0.01) and sometimes their morphology was significantly altered. Our data indicate that CK aggregates affect not only cell morphology but also the localization of various cytosolic components, which may affect the cellular function. PMID- 15748893 TI - Kinesin is involved in protecting nascent microtubules from disassembly after recovery from nocodazole treatment. AB - Upon recovery from nocodazole treatment, microtubules from cultured epithelial cells exhibit unusual properties: they re-grow as fast as any highly dynamic microtubule, but they are also protected against disassembly when challenged with nocodazole like the stable microtubules of steady-state cells. Exploring the mechanism that underlies this protection, we found that it was sensitive to ATP treatment and that it involved conventional kinesin. Kinesin localized at the growing end or along nascent microtubules. Its inhibition using a dominant negative construct for cargo binding, or by micro-injecting an anti-kinesin heavy chain antibody that impairs motor activity, resulted in the partial or total loss of microtubule protection. Finally, in an ex vivo elongation assay, we found that kinesin also participates in the control of microtubule re-growth. Altogether, our findings suggest that kinesin is involved in an early microtubule protection process that is linked to the control of their dynamics during their early growth phase. PMID- 15748894 TI - Matrix metalloproteinase inhibition impairs the processing, formation and mineralization of dental tissues during mouse molar development. AB - Organotypic cultures of embryonic mouse tooth germs were used to investigate the developmental expression and roles of MMPs in the formation and mineralization of dentin and enamel matrices. The spatially and temporally regulated expression of MMP-2, MMP-9 and MMP-20 in developing mouse tooth germs in vivo was maintained in culture. The inhibition of metalloproteinases activity by marimastat altered the morphogenesis and mineralization of the tooth germs associated with an inhibition of the activation of both MMP-20 and MMP-2. MMP inhibition deregulated the molecular processing of two major dental matrix proteins, amelogenin and dentin sialoprotein (DSP). This coincided with their accumulation and the loss of their normal distribution within the extracellular matrix, resulting in a defective mineralization of dentin and enamel matrices. These findings demonstrate the critical role of MMPs in the processing and maturation of the dental matrix. PMID- 15748895 TI - Myosin phosphatase targeting subunit 1 affects cell migration by regulating myosin phosphorylation and actin assembly. AB - Myosin II plays important roles in many contractile-like cell functions, including cell migration, adhesion, and retraction. Myosin II is activated by regulatory light chain (RLC) phosphorylation whereas RLC dephosphorylation by myosin light chain phosphatase containing a myosin phosphatase targeting subunit (MYPT1) leads to myosin inactivation. HeLa cells contain MYPT1 in addition to a newly identified human variant 2 containing an internal deletion. RLC dephosphorylation, cell migration, and adhesion were inhibited when either or both MYPT1 isoforms were knocked down by RNA interference. RLC was highly phosphorylated (60%) when both isoforms were suppressed by siRNA treatment relative to control cells (10%) with serum-starvation and ROCK inhibition. Prominent stress fibers and focal adhesions were associated with the enhanced RLC phosphorylation. The reintroduction of MYPT1 or variant 2 in siRNA-treated cells decreased stress fibers and focal adhesions. MYPT1 knockdown also led to an increase of F-actin relative to G-actin in HeLa cells. The myosin inhibitor blebbistatin did not inhibit this effect, indicating MYPT1 likely affects actin assembly independent of RLC phosphorylation. Proper expression of MYPT1 or variant 2 is critical for RLC phosphorylation and actin assembly, thus maintaining normal cellular functions by simultaneously controlling cytoskeletal architecture and actomyosin activation. PMID- 15748897 TI - The novel tumor suppressor p33ING2 enhances UVB-induced apoptosis in human melanoma cells. AB - The roles of p33ING2 as a tumor suppressor candidate have been shown through regulation of gene transcription, induction of cell cycle arrest, and apoptosis. As p33ING2 shares 58.9% homology with p33ING1b, we hypothesized that p33ING2 shares functional similarities with p33ING1b. We previously found that p33ING1b cooperates with p53 to enhance UVB-induced apoptosis. Here, we report that overexpression of p33ING2 enhanced apoptosis in UVB-irradiated and non-irradiated melanoma MMRU cells. We demonstrate that enhancement of apoptosis by p33ING2 requires the presence of functional p53. Furthermore, we found that overexpression of p33ING2 significantly downregulated the expression of Bcl-2 after UVB irradiation, resulting in an increased Bax/Bcl-2 ratio. Moreover, we found that p33ING2 promoted Bax translocation to mitochondria, altered the mitochondrial membrane potential, and induced cytochrome c release and thus the activation of caspases 9 and 3. In addition, we showed that under non-stress conditions p33ING2 upregulates Fas expression and activates caspase 8. Taken together, we demonstrate that p33ING2 cooperates with p53 to regulate apoptosis via activation of both the mitochondrial/intrinsic and death-receptor/extrinsic apoptotic pathways. PMID- 15748896 TI - Interferon-beta-induced activation of c-Jun NH2-terminal kinase mediates apoptosis through up-regulation of CD95 in CH31 B lymphoma cells. AB - Type I interferon (IFN)-induced antitumor action is due in part to apoptosis, but the molecular mechanisms underlying IFN-induced apoptosis remain largely unresolved. In the present study, we demonstrate that IFN-beta induced apoptosis and the loss of mitochondrial membrane potential (delta psi m) in the murine CH31 B lymphoma cell line, and this was accompanied by the up-regulation of CD95, but not CD95-ligand (CD95-L), tumor necrosis factor (TNF), or TNF-related apoptosis inducing ligand (TRAIL). Pretreatment with anti-CD95-L mAb partially prevented the IFN-beta-induced loss of delta psi m, suggesting that the interaction of IFN beta-up-regulated CD95 with CD95-L plays a crucial role in the induction of fratricide. IFN-beta induced a sustained activation of c-Jun NH2-terminal kinase 1 (JNK1), but not extracellular signal-regulated kinases (ERKs). The IFN-beta induced apoptosis and loss of delta psi m were substantially compromised in cells overexpressing a dominant-negative form of JNK1 (dnJNK1), and it was slightly enhanced in cells carrying a constitutively active JNK construct, MKK7-JNK1 fusion protein. The IFN-beta-induced up-regulation of CD95 together with caspase 8 activation was also abrogated in the dnJNK1 cells while it was further enhanced in the MKK7-JNK1 cells. The levels of cellular FLIP (c-FLIP), competitively interacting with caspase-8, were down-regulated by stimulation with IFN-beta but were reversed by the proteasome inhibitor lactacystin. Collectively, the IFN-beta induced sustained activation of JNK mediates apoptosis, at least in part, through up-regulation of CD95 protein in combination with down-regulation of c-FLIP protein. PMID- 15748898 TI - Tumor necrosis factor-alpha stimulates HIV-1 production in primary culture of human adipocytes. AB - Adipose tissue of HIV-1-infected patients shows severe abnormalities such as profound changes in adipose tissue morphology and metabolism. Does HIV-1 infect the adipose cell remains an unsolved question since previous attempts showed that HIV-1 poorly infects human adipocytes in vitro. In the present study, preadipose cells from human subcutaneous fat pads were differentiated in vitro, checked for HIV receptor expression, then infected with R5 and X4 HIV1 strains. Using a sensitive RT-PCR assay, we showed that HIV-1 tat and rev early viral transcripts were expressed in infected adipocytes giving a clear evidence of HIV-1 transcriptional activity in these cells. However, at the same time, no sign of productive infection was demonstrated since infected adipocytes did not efficiently produce Gag p24 antigen. We hypothesized that such a limitation could result from the lack of activation of adipocyte-signaling pathways able to stimulate HIV-1 gene expression in quiescent adipocytes. Indeed, a significant increase in Gag p24 production was observed after stimulation of infected adipocytes with pro-inflammatory cytokines, such as tumor necrosis factor alpha or interleukin-1-beta. Taken together, these results demonstrate that HIV-1 does infect human adipose cells in vitro and suggest that the initial limited infection can be overcome upon pro-inflammatory cytokine treatment. PMID- 15748899 TI - Early BrdU-responsive genes constitute a novel class of senescence-associated genes in human cells. AB - We identified genes that immediately respond to 5-bromodeoxyuridine (BrdU) in SUSM-1, an immortal fibroblastic line, with DNA microarray and Northern blot analysis. At least 29 genes were found to alter gene expression greater than twice more or less than controls within 36 h after addition of BrdU. They took several different expression patterns upon addition of BrdU, and the majority showed a significant alteration within 12 h. When compared among SUSM-1, HeLa, and TIG-7 normal human fibroblasts, 19 genes behaved similarly upon addition of BrdU. In addition, 14 genes, 9 of which are novel as regards senescence, behaved similarly in senescent TIG-7 cells. The genes do not seem to have a role in proliferation or cell cycle progression. These results suggest that the early BrdU-responsive genes represent early signs of cellular senescence and can be its new biomarkers. PMID- 15748900 TI - Dynamic reorganization of chemokine receptors, cholesterol, lipid rafts, and adhesion molecules to sites of CD4 engagement. AB - T cell polarization and redistribution of cellular components are critical to processes such as activation, migration, and potentially HIV infection. Here, we investigate the effects of CD4 engagement on the redistribution and localization of chemokine receptors, CXCR4 and CCR5, adhesion molecules, and lipid raft components including cholesterol, GM1, and glycosyl-phosphatidylinositol (GPI) anchored proteins. We demonstrate that anti-CD4-coated beads (alpha CD4-B) rapidly induce co-capping of chemokine receptors as well as GPI-anchored proteins and adhesion molecules with membrane cholesterol and lipid rafts on human T cell lines and primary T cells to the area of bead-cell contact. This process was dependent on the presence of cellular cholesterol, cytoskeletal reorganization, and lck signaling. Lck-deficient JCaM 1.6 cells failed to cap CXCR4 or lipid rafts to alpha CD4-B. Biochemical analysis reveals that CXCR4 and LFA-1 are recruited to lipid rafts upon CD4 but not CD45 engagement. Furthermore, we also demonstrate T cell capping of both lipid rafts and chemokine receptors at sites of contact with HIV-infected cells, despite the binding of an HIV inhibitory mAb to CXCR4. We conclude that cell surface rearrangements in response to CD4 engagement may serve as a means to enhance cell-to-cell signaling at the immunological synapse and modulate chemokine responsiveness, as well as facilitate HIV entry and expansion by synaptic transmission. PMID- 15748901 TI - Multi-lobulation of the nucleus in prolonged S phase by nuclear expression of Chk tyrosine kinase. AB - Chk tyrosine kinase phosphorylates Src-family tyrosine kinases and suppresses their kinase activity. We recently showed that Chk localizes to the nucleus as well as the cytoplasm and inhibits cell proliferation. To investigate the role of nuclear Chk in proliferation, various Chk mutants were constructed and expressed. Nuclear localization of Chk-induced dynamic multi-lobulation of the nucleus and prolonged S phase of the cell cycle. The N-terminal domain of Chk and a portion of its kinase domain but not the kinase activity were responsible for induction of the multi-lobulation. Cell sorting analysis revealed that nuclear multi lobulated cells were enriched in late S phase. Multi-lobulated nuclei were surrounded with lamin B1 that was particularly concentrated in concave regions of the nuclei. Furthermore, treatment with nocodazole or taxol disrupted multi lobulation of the nucleus. These results suggest that nuclear multi-lobulation in late S phase, which is dependent on polymerization and depolymerization of microtubules, may be involved in nuclear Chk-induced inhibition of proliferation. PMID- 15748902 TI - Both lamin A and lamin C mutations cause lamina instability as well as loss of internal nuclear lamin organization. AB - We have applied the fluorescence loss of intensity after photobleaching (FLIP) technique to study the molecular dynamics and organization of nuclear lamin proteins in cell lines stably transfected with green fluorescent protein (GFP) tagged A-type lamin cDNA. Normal lamin A and C proteins show abundant decoration of the inner layer of the nuclear membrane, the nuclear lamina, and a generally diffuse localization in the nuclear interior. Bleaching studies revealed that, while the GFP-tagged lamins in the lamina were virtually immobile, the intranuclear fraction of these molecules was partially mobile. Intranuclear lamin C was significantly more mobile than intranuclear lamina A. In search of a structural cause for the variety of inherited diseases caused by A-type lamin mutations, we have studied the molecular organization of GFP-tagged lamin A and lamin C mutants R453W and R386K, found in Emery-Dreifuss muscular dystrophy (EDMD), and lamin A and lamin C mutant R482W, found in patients with Dunnigan type familial partial lipodystrophy (FPLD). In all mutants, a prominent increase in lamin mobility was observed, indicating loss of structural stability of lamin polymers, both at the perinuclear lamina and in the intranuclear lamin organization. While the lamin rod domain mutant showed overall increased mobility, the tail domain mutants showed mainly intranuclear destabilization, possibly as a result of loss of interaction with chromatin. Decreased stability of lamin mutant polymers was confirmed by flow cytometric analyses and immunoblotting of nuclear extracts. Our findings suggest a loss of function of A type lamin mutant proteins in the organization of intranuclear chromatin and predict the loss of gene regulatory function in laminopathies. PMID- 15748903 TI - FoxA1 binding to the MMTV LTR modulates chromatin structure and transcription. AB - Novel binding sites for the forkhead transcription factor family member Forkhead box A (FoxA), previously referred to as Hepatocyte Nuclear Factor 3 (HNF3), were found within the mouse mammary tumor virus long terminal repeat (MMTV LTR). The effect of FoxA1 on MMTV LTR chromatin structure, and expression was evaluated in Xenopus laevis oocytes. Mutagenesis of either of the two main FoxA binding sites showed that the distal site, -232/-221, conferred FoxA1-dependent partial inhibition of glucocorticoid receptor (GR) driven MMTV transcription. The proximal FoxA binding segment consisted of two individual FoxA sites at -57/-46 and -45/-34, respectively, that mediated an increased basal MMTV transcription. FoxA1 binding altered the chromatin structure of both the inactive- and the hormone-activated MMTV LTR. Hydroxyl radical foot printing revealed FoxA1 mediated changes in the nucleosome arrangement. Micrococcal nuclease digestion showed the hormone-dependent sub-nucleosome complex, containing approximately 120 bp of DNA, to be expanded by FoxA1 binding to the proximal segment into a larger complex containing approximately 200 bp. The potential function of the FoxA1 mediated expression of the MMTV provirus for maintenance of expression in different tissues is discussed. PMID- 15748905 TI - Scanning electron microscopy study of neutrophil membrane tubulovesicular extensions (cytonemes) and their role in anchoring, aggregation and phagocytosis. The effect of nitric oxide. AB - We have shown that human neutrophils develop dynamic thin and very long tubulovesicular extensions (cytonemes) upon adhesion to fibronectin, if cell spreading was blocked by Na(+)-free medium or by 4-bromophenacyl bromide, N ethylmaleimide, 7-chloro-4-nitrobenz-2-oxa-1,3-diazole and cytochalasin D (S. I. Galkina, G. F. Sud'ina and V. Ullrich, (2001). Exp. Cell Res. 266, 222-228). In the present work we found that similar in size and behavior tubulovesicular extensions were formed on the neutrophil cell bodies upon adhesion to fibronectin coated substrata in the presence of the nitric oxide donor diethylamine NONOate. In the presence of the nitric oxide synthase inhibitor N-omega-nitro-L-arginine methyl ester, neutrophils were well spread and had no microextensions. Using scanning electron microscopy, we demonstrated that tubulovesicular extensions of neutrophils executed long-range adhesion and binding objects for phagocytosis, such as serum-opsonized zymosan particles and erythrocytes. Tubulovesicular extensions anchored neutrophils to substrata in a beta1 and beta2 integrin independent, but L-selectin-dependent manner. BODIPY-sphingomyelin impaired development of tubulovesicular extension, and heparitinase 1 played a role in their destruction. Membrane tubulovesicular extensions are supposed to represent protrusions of an intracellular exocytotic traffic and serve as cellular sensory and adhesive organelles. Nitric oxide seems to play a role in regulation of tubulovesicular extensions formation, thus affecting neutrophil adhesive interactions and phagocytosis. PMID- 15748904 TI - Epidermal growth factor receptor coexpression modulates susceptibility to Herceptin in HER2/neu overexpressing breast cancer cells via specific erbB receptor interaction and activation. AB - BACKGROUND: Growth factors and Herceptin specifically and differentially modulate cell proliferation of tumor cells. However, the mechanism of action on erbB receptor level is incompletely understood. We evaluated Herceptin's capacity to modulate erbB-receptor activation and interaction on the cell surface level and thereby potentially impair cell proliferation of HER2/neu (c-erbB2) overexpressing breast cancer cells, both in the presence and absence of relevant growth factors. METHODS: BT474 and SK-BR-3 breast cancer cell lines were treated with Epidermal Growth Factor (EGF), Heregulin, and with Herceptin in different combinations. Kinetics of cell proliferation were evaluated flow cytometrically based on BrdU-labeling. Fluorescence Resonance Energy Transfer, ELISAs and phosphorylation site specific Western Blotting was performed to investigate erbB receptor interaction and activation. RESULTS: EGF induced EGFR/EGFR and EGFR/c erbB2 interactions correlate with stimulation of cell proliferation in BT474 cells. Both homo- and heterodimerization are considerably less pronounced in SK BR-3 cells and heterointeraction is additionally reduced by EGF treatment, causing inhibition of cell proliferation. Heregulin stimulates cell proliferation extensively in both cell lines. Herceptin drives BT474 cells more efficiently into quiescence than it does with SK-BR-3 cells and thereby blocks cell cycle progress. In SK-BR-3 Herceptin treatment causes c-erbB2 phosphorylation of Y877 and Y1248, EGF induces Y877 and Y1112 phosphorylation. The Y1112 phosphorylation site, activated by EGF in SK-BR-3 cell, is bypassed in BT474. In addition the inhibitory capacity of Herceptin on BT474 and SK-BR-3 cell proliferation depends on the presence and absence of growth factors to a various extent. CONCLUSION: The growth inhibitory effect of Herceptin on c-erbB2 overexpressing breast cancer cells is considerably modulated by EGFR coexpression and consequently EGFR/c erbB2 homo- and heterointeractions, as well as the presence or absence of growth factors. C-erbB2 overexpression alone is insufficient to predict the impact of growth factors and antibodies on cell proliferation. The optimization and specification of therapeutic approaches based on erbB-receptor targeting requires to account for EGFR coexpression as well as the potential presence of erbB receptor relevant growth factors. PMID- 15748906 TI - Ligand depletion negatively controls the mitogenic activity of epidermal growth factor. AB - EGF activates the ErbB1 receptor, but there appears only a limited correlation between its receptor binding affinity and mitogenic activity. This is indicated by our present observation that in cells with high ErbB1 expression, including SUM102 breast tumor cells, low affinity EGF/Notch chimeras have similarly high mitogenic activity as EGF, in spite of the fact that EGF is superior in inducing receptor tyrosine phosphorylation and p42/p44 MAP-kinase activity. However, as a result of receptor-mediated internalisation high-affinity ligands such as EGF are depleted much more rapidly from the extracellular medium than low-affinity EGF/Notch chimeras. As a consequence, the mitogenic activity of EGF on ErbB1 overexpressing cells is limited by substantial degradation of internalised ligand in the period before cells enter S-phase, a phenomenon that is not observed for low affinity mutant ligands. The mitogenic activity of EGF on ErbB1 overexpressing cells does therefore not only depend on the applied concentration but also on the total amount of ligand added, and is strongly underestimated when tested in a limited assay volume. No such dependence on the incubation volume was observed for EGF activity on cells with low ErbB1 expression levels and on cells for which EGF is growth inhibitory. PMID- 15748907 TI - Food toxicity contributes to the beneficial effects of calorie restriction. PMID- 15748908 TI - Temperature compensation in circadian clock models. AB - Circadian clock of organisms has a free-running period that does not change much with ambient temperature. This property "temperature compensation" is studied when the rate of all reaction steps increase with temperature in the biochemical network generating the rhythm. The period becomes shorter when all the rate parameters are enhanced by the same factor. However, the period becomes longer as degradation rate of proteins and/or transcription rate of the clock gene increase (their elasticity is positive). This holds for a wide range of models, including N-variable model, and PER-TIM double oscillator model, provided that (1) branch reactions (e.g. degradation of proteins or mRNAs) are strongly saturated, and that (2) the cooperativity of transcription inhibition by nuclear proteins is not very large. A strong temperature sensitivity of degradation of PER proteins and/or temperature-sensitive alternative splicing of per gene, known for Drosophila, can be mechanisms for the temperature compensation of circadian clock. PMID- 15748909 TI - Simulating the impact of a molecular 'decision-process' on cellular phenotype and multicellular patterns in brain tumors. AB - Experimental evidence indicates that human brain cancer cells proliferate or migrate, yet do not display both phenotypes at the same time. Here, we present a novel computational model simulating this cellular decision-process leading up to either phenotype based on a molecular interaction network of genes and proteins. The model's regulatory network consists of the epidermal growth factor receptor (EGFR), its ligand transforming growth factor-alpha (TGF alpha), the downstream enzyme phospholipaseC-gamma (PLC gamma) and a mitosis-associated response pathway. This network is activated by autocrine TGF alpha secretion, and the EGFR dependent downstream signaling this step triggers, as well as modulated by an extrinsic nutritive glucose gradient. Employing a framework of mass action kinetics within a multiscale agent-based environment, we analyse both the emergent multicellular behavior of tumor growth and the single-cell molecular profiles that change over time and space. Our results show that one can indeed simulate the dichotomy between cell migration and proliferation based solely on an EGFR decision network. It turns out that these behavioral decisions on the single cell level impact the spatial dynamics of the entire cancerous system. Furthermore, the simulation results yield intriguing experimentally testable hypotheses also on the sub-cellular level such as spatial cytosolic polarization of PLC gamma towards an extrinsic chemotactic gradient. Implications of these results for future works, both on the modeling and experimental side are discussed. PMID- 15748911 TI - Selection of initial conditions for recursive production of multicellular organisms. AB - The development of a multicellular organism is a dynamic process. Starting from one or a few cells, the organism becomes a set of cells with different types that form well-determined patterns. It is rather surprising that differentiation in cell types and formation of controlled patterns are compatible, because the former gives morphogenetic diversification whereas the latter implies recursive production of a cell ensemble, reducing individual differences. We studied this problem by taking a simple cell model with intracellular reaction dynamics of chemical concentrations, cell-cell interactions, and increase in cell numbers. We observed successive differentiation from a cell type with diverse chemicals and chaotic concentration dynamics to cell types with oscillatory or fixed-point dynamics, leading to morphogenetic diversity in a spatial pattern. We further show that, by starting from an initial object consisting of both the former cell type with diverse chemicals and the latter differentiated cell type, the recursive production of a multicellular organism with morphogenetic diversity is possible. By relating the former type to a cell in the vegetal pole and the latter to one in the animal pole, classic experimental results with separation of blastomeres in sea urchin eggs are coherently explained, while some predictions are made for in vitro morphogenesis from embryonic stem cells. PMID- 15748910 TI - The kinetics of FGF-2 binding to heparan sulfate proteoglycans and MAP kinase signaling. AB - Binding of growth factors to specific cell surface receptors is the first step in initiating cell signaling cascades that ultimately result in diverse activities such as proliferation, differentiation, and apoptosis. Dimerization and phosphorylation of tyrosine kinase transmembrane receptors is the typical paradigm for this activation but, for many growth factors, cell surface interactions are not limited to a single receptor type. In particular, heparin binding growth factors, such as fibroblast growth factor-2 (FGF-2), bind to heparan sulfate proteoglycans (HSPG) on the cell surface and within the extracellular matrix (ECM), and these molecules have been viewed as accessory co receptors serving to facilitate tyrosine kinase receptor binding. Recent studies, however, have indicated that HSPG can directly participate in signal transduction in response to FGF-2 binding. Thus, in the present study, we used mathematical modeling to examine whether the kinetics of formation of the various FGF-2 bound complexes on the cell surface correlate with the activation of the downstream mediators of FGF-2 response, Erk1/2. We find that FGF-2 binding to its receptor correlates well with Erk1/2 activation and that HSPG can modulate this response through its ability to stabilize these ligand receptor complexes. Moreover, we also observed that FGF-2 binding to HSPG correlates strongly with Erk1/2 activation under conditions where there is a loss of receptor activity, and we demonstrate that the relative amounts of signaling and non-signaling HSPG on the cell surface, as well as the presence of competing HSPG in the ECM, can impact the signal potential via this pathway. Thus, the selective regulation of specific HSPG might provide a mechanism for fine tuned modulation of heparin-binding growth factor signaling in cells where signal intensity and duration could direct cellular response toward growth, migration or differentiation. PMID- 15748912 TI - Scale-free neurodegeneration: cellular heterogeneity and the stretched exponential kinetics of cell death. AB - Neurodegenerative disorders are an insidious group of diseases characterized by severe physical and cognitive effects that often have devastating consequences for the lives of affected individuals and their families. One feature common to a significant proportion of these diseases is that affected neurons commit to undergoing an active form of degeneration known as programmed cell death, or apoptosis. Although intense effort over the past several years has resulted is a remarkable increase in our understanding of the molecular events involved in neurodegeneration, our knowledge regarding the cellular and tissue properties that determine the temporal patterns of neuronal attrition is limited. We recently demonstrated that neurodegenerative kinetics in various diseases fit well to exponential decay functions, and proposed a universal one-hit switch mechanism in which mutant and injured neurons exist in a viable state characterized by an increased but constant risk of initiating apoptosis (Nature, 406, p. 195). Here we show that a heavy-tailed stretched exponential function is better able to account for neurodegenerative kinetic data. Moreover, normalization of all available data according to their corresponding best-fit stretched exponential parameters suggest that the generalized model is consistent with a universal mechanism of neuronal cell death that is greatly improved over the constant risk model. In contrast to the original model in which all cells exhibit an identical risk of initiating apoptosis, the stretched exponential model is consistent with each neuron experiencing a constant risk that is different from that experienced by other cells in the degenerating population, perhaps due to spatial differences in the cellular microenvironment. Intriguingly, the predicted distribution of risk across the cell population can be fit by a power-law function, further suggesting that scale-free properties of degenerating neuronal tissues might act as potent regulators of the kinetics of cell death in neural tissue. PMID- 15748913 TI - The triplet genetic code had a doublet predecessor. AB - Information theoretic analysis of genetic languages indicates that the naturally occurring 20 amino acids and the triplet genetic code arose by duplication of 10 amino acids of class-II and a doublet genetic code having codons NNY and anticodons GNN. Evidence for this scenario is presented based on the properties of aminoacyl-tRNA synthetases, amino acids and nucleotide bases. PMID- 15748914 TI - Quantifying macrophage defects in type 1 diabetes. AB - Macrophages from animals prone to autoimmune (type 1) diabetes differ from those of diabetes-resistant animals in processing and clearing apoptotic cells. Using in vitro time-course assays of the number of engulfed apoptotic cells observed within macrophages, we quantified these differences in non-obese diabetic (NOD) versus Balb/c mice. Simple models lead to several elementary parameter estimation techniques. We used these to compute approximate rates of macrophage engulfment and digestion of apoptotic cells from basic features of the data (such as initial rise-times, phagocytic index and percent phagocytosis). Combining these estimates with full fitting of a sequence of model variants to the data, we find that macrophages from normal (Balb/c) mice engulf apoptotic cells up to four times faster than macrophages from the diabetes-prone (NOD) mice. Further, Balb/c macrophages appear to undergo an activation step before achieving their high engulfment rate. In NOD macrophages, we did not see evidence for this activation step. Rates of digestion of engulfed apoptotic cells by macrophages are similar in both types. Since macrophage clearance is an important mechanism of disposal of self-antigen, these macrophage defects could potentially be a factor in predisposition to type 1 diabetes. PMID- 15748915 TI - Recurrent epidemics in small world networks. AB - The effect of spatial correlations on the spread of infectious diseases was investigated using a stochastic susceptible-infective-recovered (SIR) model on complex networks. It was found that in addition to the reduction of the effective transmission rate, through the screening of infectives, spatial correlations have another major effect through the enhancement of stochastic fluctuations, which may become considerably larger than in the homogeneously mixed stochastic model. As a consequence, in finite spatially structured populations significant differences from the solutions of deterministic models are to be expected, since sizes even larger than those found for homogeneously mixed stochastic models are required for the effects of fluctuations to be negligible. Furthermore, time series of the (unforced) model provide patterns of recurrent epidemics with slightly irregular periods and realistic amplitudes, suggesting that stochastic models together with complex networks of contacts may be sufficient to describe the long-term dynamics of some diseases. The spatial effects were analysed quantitatively by modelling measles and pertussis, using a susceptible-exposed infective-recovered (SEIR) model. Both the period and the spatial coherence of the epidemic peaks of pertussis are well described by the unforced model for realistic values of the parameters. PMID- 15748916 TI - Reduction of intrinsic kinetic and thermodynamic barriers for enzyme-catalysed proton transfers from carbon acid substrates. AB - Many enzymes catalyse the heterolytic abstraction of the alpha-proton from a carbon acid substrate. Gerlt and Gassman have applied Marcus formalism to such proton transfer reactions to argue that transition states for concerted general acid-general base catalysed enolization at enzyme active sites occur late on the reaction coordinate (J. Am. Chem. Soc. 115 (1993) 11552). We postulate that as an enzyme evolves, it may decrease deltaG++ for a proton transfer step associated with substrate enolization by following the path of steepest descent on the two dimensional surface corresponding to deltaG++, as defined by Marcus formalism. We show that for an enzyme that has decreased deltaG++ following the path of steepest descent, the values of the intrinsic kinetic (deltaG++(int,E)) and thermodynamic (deltaG(E)0) barriers for proton transfer reactions on the enzyme may be predicted from the known values of deltaG++(int,N) and deltaG(N)0 for the corresponding non-enzymic reaction and the free energy of activation on the enzyme (deltaG++(E)). In addition, the enzymic transition state will occur later on the reaction coordinate than the corresponding non-enzymic transition state (i.e. x++(E)>x++(N)) if the condition (6 - square root 2)/82deltaG++(int,N). PMID- 15748917 TI - Sampling rate effects on measurements of correlated and biased random walks. AB - When observing the two-dimensional movement of animals or microorganisms, it is usually necessary to impose a fixed sampling rate, so that observations are made at certain fixed intervals of time and the trajectory is split into a set of discrete steps. A sampling rate that is too small will result in information about the original path and correlation being lost. If random walk models are to be used to predict movement patterns or to estimate parameters to be used in continuum models, then it is essential to be able to quantify and understand the effect of the sampling rate imposed by the observer on real trajectories. We use a velocity jump process with a realistic reorientation model to simulate correlated and biased random walks and investigate the effect of sampling rate on the observed angular deviation, apparent speed and mean turning angle. We discuss a method of estimating the values of the reorientation parameters used in the original random walk from the rediscretized data that assumes a linear relation between sampling time step and the parameter values. PMID- 15748920 TI - College health. PMID- 15748918 TI - Engineers have more sons, nurses have more daughters: an evolutionary psychological extension of Baron-Cohen's extreme male brain theory of autism. AB - In his extreme male brain theory of autism, Baron-Cohen postulates that having a typically male brain was adaptive for ancestral men and having a typically female brain was adaptive for ancestral women. He also suggests that brain types are substantially heritable. These postulates, combined with the insight from the Trivers-Willard hypothesis regarding parental ability to vary offspring sex ratio, lead to the prediction that people who have strong male brains should have more sons than daughters, and people who have strong female brains should have more daughters than sons. The analysis of the 1994 US General Social Survey data provides support for this prediction. Our results suggest potentially fruitful extensions of both Baron-Cohen's theory and the Trivers-Willard hypothesis. PMID- 15748921 TI - A case study on the use of physician assistants in college health. AB - This article describes how the use of physician assistants can maximize high quality, cost-effective patient care. In addition to serving as staff members of the health care team and in administrative capacities, physician assistants can serve in a number of functions in college health. PMID- 15748922 TI - Common medical problems of the college student. AB - The college health physician cares for college students who present with a wide variety of medical disorders. This article reviews the management of four common medical problems: infectious mononucleosis, asthma, migraine headaches, and urinary tract infections. College students can become ill, and it is important that they have health care services designed to deal with their health care issues. PMID- 15748923 TI - The college athlete. AB - Participation in sports is important to many college students. Student athletes come from different levels of previous sport experience as they enter collegiate athletics. The primary source of student medical care is the campus student health center. The health care providers at student health centers attend to many of the sports-related concerns of student athletes. Preparticipation evaluation provides an opportunity to assess the general health of the student athlete and to identify conditions that might increase the risk of further injury. Sudden cardiac death and sports-associated concussions have generated much interest and are reviewed in this article. Other areas reviewed here include use of drugs and supplements, ankle sprains, acute knee ligament injuries, back pain, and shoulder impingement syndrome. PMID- 15748924 TI - Understanding college students with learning disabilities. AB - The percentage of full-time college freshmen reporting disabilities at 4-year institutions remained stable (6-8%) between 1988 and 2000. By 2000, two in five freshmen with disabilities (40%) reported having a learning disability. In 2002, learning disabilities accounted for 51% of special education classifications. The effects of chronic school failure frequently shape the experience of students with learning disabilities. This article reviews common learning disabilities found in college students and provides suggestions for helping these students. PMID- 15748925 TI - Attention-deficit/hyperactivity disorder psychopharmacology for college students. AB - This article is written as a practical guide for the use of medications in the management of attention-deficit/hyperactivity disorder in college students. It includes a review of the common medications used and an algorithm of treatment using these medications. Practical guidelines are presented along with comments regarding more common side effects. The article also addresses some concerns about abuse of psychostimulants by college students. PMID- 15748926 TI - Eating disorders in college. AB - Anorexia nervosa and bulimia nervosa are serious chronic psychiatric disorders that can result in significant medical and psychologic outcomes. These multifaceted disorders affect the emotions, thinking, behavior, and physical health of afflicted individuals. Symptoms often peak during the years many youth are attending college. The intersection of issues of emancipation, individuation, intimacy, and eating disorders may be part of the reason that researchers report a high incidence of eating disorders in this specialized population. This article presents an overview of eating disorders in the college population and covers psychologic and psychopharmacologic treatment. PMID- 15748927 TI - Anxiety and depressive disorders in college youth. AB - Fear and anxiety are common experiences throughout childhood and adolescence. Anxiety disorders, along with depression and dysthymia, are characterized as internalizing disorders; they stand in distinction from the externalizing disorders representing such conditions as oppositional-defiant disorder and conduct disorder. Clinicians evaluating anxiety disorders in college youth face the task of differentiating normal, transient, developmentally appropriate expression of anxiety from pathologic states of anxiety. The developmental course of anxiety and depression, its appropriateness, and its boundaries are areas of research and interest. A wide clinical perspective is necessary to effectively engage the assessment process. The treatment of anxiety and depression in youth is ideally multi-modal, involving medication, psychotherapy, and psychosocial interventions. This article focuses on assessment and treatment of anxiety and depression. These are presented separately, although areas of overlap often are encountered in practice. PMID- 15748928 TI - Contraception for college students. AB - An important issue for human beings is the acquisition of normal sexual health, including the understanding and application of reproductive health when needed. Comprehensive sexuality education is not a topic provided to many children, adolescents, or college students in the United States, however. Students who were sexually active in high school may continue to be at risk for unwanted pregnancies and sexually transmitted diseases in their college life; those who chose abstinence in high school may abandon this concept in college, choosing coital behavior at all levels of university life-freshman through graduate levels. Most American college students are sexually active and many have multiple partners. This article reviews current contraceptive methods available to college students. College health providers and pediatricians are urged to be vigilant about the reproductive health needs of the college students they serve. PMID- 15748929 TI - Human papilloma virus, papanicolaou smears, and the college female. AB - This article discusses human papilloma virus (HPV), Papanicolaou smears and the rational for new screening guidelines. PMID- 15748930 TI - Menstrual disorders in the college age female. AB - College-age young women frequently experience a variety of menstrual-related complaints, including dysmenorrheal, menorrhagia, irregular menses, and menstrual related mood changes. These problems deserve careful evaluation; they may reflect normal ovulatory menstrual symptoms or be suggestive of significant pathology that can have a major impact on future reproductive and general health. The menstrual cycle is a vital sign whose normalcy suggests an overall good health and whose abnormality requires evaluation. Eating disorders and the female athlete triad increase the risk of osteoporosis; polycystic ovary syndrome is associated with future cardiovascular risks. Diagnosis and management of these problems will not only improve a young woman's current health, sense of well being, and overall quality of life but may also lower her risks for future disease and ill-health. This article addresses normal menstrual function, excessive bleeding, infrequent or absent menses, pain with menses, menstrual related mood disorders, and recommendations about routine gynecologic examinations and evaluation. PMID- 15748931 TI - Genitourinary issues in the male college student: a case-based approach. AB - Five percent of all ambulatory visits by men 18 years of age or older include genitourinary symptoms as a reason for the visit. In this article, using typical, unusual, or otherwise instructive cases, the authors review a select group of genitourinary issues in the college-age male. Warts (human papilloma virus), is the most common sexually transmitted infection, and it may mimic other disease. Testicular cancer is one of the most serious diseases to confront health care providers. Varicoceles are the most common scrotal mass. Urethritis is a common presentation of sexually transmitted infection in the young adult male. Acute prostatitis is an unusual condition in the young adult, but it is easily treatable. Sexual dysfunction causes great distress in the young adult, but a systematic approach usually leads to a treatable psychological or environmental cause. With understanding of these medical conditions, the practitioner should feel comfortable addressing the most challenging genitourinary health needs of this population. PMID- 15748932 TI - Sexually transmitted infections: new guidelines for an old problem on the college campus. AB - Approximately 80% of college-age adolescents are sexually active and at risk for sexually transmitted infections (STIs). Over 4 million STIs occur in teenagers annually and young adults between the ages of 18 and 24, while adolescents 15 to 17 years of age have higher rates of STIs than any other age group in the United States. Thus, the prevention, diagnosis, and treatment of STIs are a critical part of college health care. This article will discuss the epidemiology, diagnosis, and management of some of the most common STIs encountered in the college-age group, with an emphasis on new guidelines for treatment. PMID- 15748933 TI - Immunizations for the college student: a campus perspective of an outbreak and national and international considerations. AB - Although vaccine-preventable diseases have declined to record-low levels in the United States, infectious disease "epidemics" on college campuses continue. A large student body with variable immunization status makes a college campus fertile ground for the spread of communicable diseases. The presence of international students and an increasingly large number of students traveling abroad make it essential that individuals charged with defining and instituting health-related policies for the university have knowledge about health issues occurring in foreign countries as well. Several safe and effective vaccines are available that offer protection to young adults from a variety of infectious diseases in the United States. Because vaccine-preventable diseases can cause both human and economic problems for colleges and universities, administrators should take steps to assure that the students on college campuses benefit from these vaccines. PMID- 15748934 TI - Cardiovascular disorders in the college student. AB - This article provides an overview of diagnosis and treatment of college students with possible congenital or acquired heart problem issues. Preventive cardiac concerns and issues regarding risk factors for atherosclerotic heart disease are discussed. College students with corrected or palliated heart conditions are included because they are an expanding segment of the adult cardiac population. Selected major syndromes with associated cardiac anomalies and complications are included. Finally, reinforcement and review of cardiac diagnosis or management issues occurring in college students (six case scenarios) are included in the appendix. PMID- 15748935 TI - Diabetes on the college campus. AB - The college campus presents a unique scenario where older adolescents and young adults find themselves in an independent environment. The students with pre existing diabetes face immense responsibility regarding their diabetes care and decision making, without the immediate presence of their parents and the pediatric diabetes team. In addition, there are many other students who may be faced with a diagnosis of diabetes mellitus first identified in college. Current diabetes management strategies offer comprehensive care, which results in improved glycemic control and near-normal lifestyle. Continued effort at comprehensive diabetes education goes a long way toward giving these students healthy lives. This article reviews issues involving care of college students with diabetes. PMID- 15748936 TI - Substance abuse on the college campus. AB - Substance abuse is a major health and behavioral concern in college students. Alcohol and marijuana are the most commonly abused drugs on college campuses. Others include tobacco, 3,4-methylenedioxymethamphetamine, gamma-hydroxybutyrate, flunitrazepam (Rohypnol), lysergic acid, ketamine, methamphetamine, phencyclidine, cocaine, and psilocybin mushrooms. This article reviews the use of these drugs by college students. Substance use is a major contributing factor in poor academic performance and failure to successfully complete a college education. PMID- 15748939 TI - Determination of the sequential degradation of myelin proteins by macrophages. AB - Demonstration of different myelin proteins (myelin basic protein [MBP], proteolipid protein [PLP] and myelin oligodendrocyte glycoprotein [MOG]) is used as a tool to determine the stage of MS lesions in autopsy tissue. Since such tissue can never be obtained at well-defined stages of lesion formation, the time course of myelin degradation in MS lesions can only be estimated. In order to obtain a more precise indication on the sequence of events of myelin degradation in MS lesions, the breakdown of human myelin by human monocytes was studied in vitro. Human monocytes were fed with myelin; next cytocentrifuge preparations were made on several time points (day 0 until day 6). The cytospots were immunocytochemically stained with mono- and polyclonal antibodies directed against various myelin proteins (MOG, MBP, PLP). We found that MOG is degraded after 1 day, whereas PLP and MBP can be detected for a longer period, 2 and 3 days, respectively. The exact time frame of myelin degradation in our in vitro assay cannot be extrapolated to the MS lesion formation in vivo, but our data allow conclusions on the sequence of events as well as a rough indication of the time frame of myelin degradation by macrophages in MS lesions. PMID- 15748938 TI - Low immunogenicity of in vitro-expanded human neural cells despite high MHC expression. AB - The ability to expand human neural precursor cells in vitro offers new possibilities for future cell therapies. However, concern over immunologically based rejection of in vitro-expanded human neural cells confounds their use as donor cells. Here, we demonstrate that the expression of human leukocyte antigen (HLA) class I and II molecules, but not the co-stimulatory proteins CD40, CD80 and CD86, substantially increase during expansion of neurospheres. Furthermore, peripheral lymphocytes were unresponsive when co-cultured with in vitro-expanded neural cells. Taken together, these results suggest a low immunogenicity of these cultured human neural cells despite HLA incompatibility and high HLA expression. PMID- 15748940 TI - Role of nociceptin/orphanin FQ (Noc/oFQ) in murine experimental colitis. AB - Nociceptin/orphanin (Noc/oFQ), endogenous agonist for nociceptin receptor (NOR), is thought to be a stimulator of neurogenic inflammation. We investigated the possible role of Noc/oFQ in the development of colitis using NOR-deficient mice treated with dextran sulfate sodium (DSS). Colitis was significantly improved in NOR-deficient mice against wild-type mice. Expression level of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) and infiltrating cells also significantly decreased in NOR-deficient mice against wild-type mice. Nociceptin expression increased in wild-type mice after DSS treatment. These results suggest stimulation by Noc/oFQ deteriorates colonic inflammation via up-regulation of adhesion molecule. PMID- 15748941 TI - Severe T-cell depletion from the PALS leads to altered spleen composition in common marmosets with experimental autoimmune encephalomyelitis (EAE). AB - Recent data suggest that the spleen is a crucial component of the immune system in the development of experimental autoimmune encephalomyelitis (EAE) in marmoset monkeys. Using immunohistochemistry, we investigated changes in the distribution of leukocytes in the spleen associated with clinical symptoms of EAE. Animals without EAE displayed well-developed T- and B-cell areas, germinal centers and red pulp. In contrast, a marked depletion of periarteriolar T cells with preservation of other elements was found in animals with clinical EAE. These findings suggest that immune responses within the spleen are impaired during a paralysing inflammatory process in the central nervous system. PMID- 15748942 TI - Delineation of the minimal encephalitogenic epitope of proteolipid protein peptide(91-110) and critical residues required for induction of EAE in HLA-DR3 transgenic mice. AB - Previously, we have reported that proteolipid protein (PLP) peptide 91-110 can induce experimental autoimmune encephalomyelitis (EAE) in HLA-DR3 transgenic (tg) mice. Here we, report that residues spanning 97-108 are the minimal epitope required for induction of EAE in DR3 mice. Utilizing a series of alanine substituted peptides, positions 99, 101, 102, 103, 104, and 106 are identified as residues necessary for an immune response. Further analysis indicated that amino acid isoleucine (99), aspartate (102) and lysine (104) are anchor residues facilitating binding to HLA-DR3 molecules. These results may have applications in the future design of peptide based immunotherapy. PMID- 15748944 TI - Diabetes increases expression of ICAM after a brief period of cerebral ischemia. AB - The objective of present study was to determine whether leukocyte-endothelial cell adhesive molecule, intercellular cell adhesion molecule-1 (ICAM-1) was increased after ischemia in diabetic rats. The immunohistochemistry of ICAM showed that numbers of ICAM-1 positively stained microvessels in the cortex were markedly increased at 3 days of reperfusion in diabetic, but not in non-diabetic rats. These were further confirmed by Western analysis. Western analyses also showed that interlukin-1beta (IL-1beta), but not TNF-alpha, was increased at 3 days of the reperfusion in diabetic rats. The results suggest that inflammatory responses may mediate diabetic hyperglycemia-aggravated brain damage induced by ischemia and reperfusion. PMID- 15748943 TI - The activation of IL-8 receptors in cultured guinea pig Muller glial cells is modified by signals from retinal pigment epithelium. AB - Interleukin 8 (IL-8, CXCL8) is a pro-inflammatory chemokine which attracts neutrophils to sites of inflammation via an activation of the G-protein-coupled receptors, CXCR1 and CXCR2. However, both IL-8 and IL-8 receptors are widely expressed in various tissues and cell types, and have been suggested to be involved in other functions such as angiogenesis, tumor growth, or brain pathology. We examined the expression of IL-8 and IL-8 receptors in highly enriched primary cultures of guinea pig Muller glial cells. Immunoreactivity for CXCL8, CXCR1 and CXCR2 was observed in all cultured Muller cells. The expression of CXCL8 was confirmed by PCR, and the secretion of the CXCL8 protein from Muller cells was revealed by ELISA. Western blots showed prominent bands at approximately 40 kDa by using antibodies specific for human CXCR1 and CXCR2, and the expression of a putative CXCR2 receptor in Muller cells was confirmed by PCR. Furthermore, cultured Muller cells responded to application of recombinant human IL-8 with an increase of the cytosolic Ca(2+) concentration. If supernatants of cultured human retinal pigment epithelium (RPE) cells were applied to the Muller cell cultures, no obvious changes were observed in the CXCL8, CXCR1 and CXCR2 expression but (i) Muller cell proliferation was stimulated, and (ii) there was an increased number of CXCL8-responsive Muller cells and the amplitudes of the evoked calcium responses were enhanced. It is concluded that Muller glial cells may participate in the inflammatory response(s) of the retina during ocular diseases, and that this contribution may be modified by interactions with RPE cells. PMID- 15748946 TI - Immunodetection of anti-MAG IgM antibody by cross-reactivity to LA-N-1 neuroblastoma cells. AB - Demyelinating polyneuropathiy associated with IgM paraproteinemia and high titers of anti-MAG IgM antibodies (MAG-PN) is considered different from chronic inflammatory demyelinating polyneuropathy, particularly because of the poorer response to treatment of MAG-PN patients. Therefore, anti-MAG anitbodies may have relevant prognostic value. Available anti-MAG antibody assays require central nervous system myelin proteins from autopsied human brains. This study investigated the feasibility of detecting anti-MAG antibody by immunofluorescence and flow cytometry using a panel of human neuroblastoma cell lines as targets. We report here on the evaluation of the LA-N-1 cell line as an appropriate substrate for the detection of anti-MAG antibody by indirect immunoflourescence. PMID- 15748945 TI - The immunostimulatory activity of CpG oligonucleotides on microglial N9 cells is affected by a polyguanosine motif. AB - Oligonucleotides (ODN) with hexameric motifs containing central unmethylated CpG dinucleotides are immunostimulatory. Also ODN with continuous guanosines (polyG motif) show a wide range of immunological activity. Depending on the position, the chemical property of the ODN backbone and the cell type, polyG motifs have either an enhancing or a suppressing effect on the immunostimulatory activity of the CpG-ODN. Microglial cells are central components of the innate immune system of the brain and are activated by CpG-ODN in vitro and in vivo. Here we present the analysis of the immunomodulatory effects of CpG-ODN carrying a polyG motif on the microglial cell line N9. Our data show that N9 cells express Toll-like receptor 9 (TLR9) and are activated by CpG-ODN, which leads to expression of interleukin-12p40 (IL12p40), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS). A 3'-end polyG motif inhibits phosphothioate (PS) CpG-ODN immunostimulatory activity but enhances the immunostimulatory activity of phosphodiester (PE) CpG-ODN. Correspondingly, a 3' end polyG motif improves the cellular uptake of PE CpG-ODN but does not change their cellular distribution pattern. Furthermore, PE CpG-ODN with a 3'-end polyG motif interact with a much higher number of cellular proteins than PE CpG-ODN. These data indicate that the 3'-end polyG motif could enhance the immunostimulatory activity of PE CpG-ODN in microglial N9 cells through increasing interaction with cellular proteins. Therefore PE CpG-ODN containing a 3'-end polyG motif resulting in increased immunostimulatory activity might be promising alternate analogues for studies in the central nervous system. PMID- 15748947 TI - Gene expression and regulation of nerve growth factor in atopic dermatitis mast cells and the human mast cell line-1. AB - The gene expression and regulation of nerve growth factor (NGF) in atopic dermatitis (AD) and the human mast cell line (HMC)-1 was investigated at the molecular level. NGF-stimulation of HMC-1 cells resulted in increases in tryptase activity and histamine contents, paralleled by an increase of tryptase and histamine at the transcriptional level. Also, an increased expression of NGF was found in AD lesions, in association with increased systemic NGF plasma levels. Further cutaneous sources for increased NGF levels were keratinocytes and fibroblasts. These findings demonstrate an increased expression of NGF in AD and effects on tryptase and histamine. Mast cells may be major mediators of neurotrophin effects in AD. PMID- 15748948 TI - Interleukin-1beta potentiates neuronal injury in a variety of injury models involving energy deprivation. AB - The purpose of this study was to develop a suitable in vitro model system to study the biochemical pathway(s) by which interleukin-1beta (IL-1beta) contributes to the pathogenesis of cerebral ischemia. Thus, the effect of IL 1beta on a number of injury paradigms associated with energy deprivation was investigated using murine mixed cortical cell cultures. While IL-1beta by itself was not neurotoxic, pre-treatment-but not concurrent or post-treatment-with this cytokine potentiated neuronal injury induced by depriving cultures of either oxygen, glucose, or both oxygen and glucose. Cytotoxicity was abolished by an IL 1beta-neutralizing antibody. Together, these results demonstrate the establishment of reliable and reproducible in vitro models that will now allow detailed investigation of the cellular and molecular mechanisms relating to IL 1beta-mediated neuronal cell death. PMID- 15748949 TI - A paradoxical role of APCs in the induction of intravenous tolerance in experimental autoimmune encephalomyelitis. AB - The central role of T cells in the induction of tolerance to autoantigens has been well documented. However, the role of antigen-presenting cells (APCs) in this process is not yet fully understood. To better understand the contribution of APCs in tolerance, we studied the interaction of purified APCs and CD4(+) T cells in a model of intravenous (i.v.) tolerance to experimental autoimmune encephalomyelitis (EAE). As expected, we found that T cells were tolerized to the autoantigen after i.v. injection. However, purified APCs obtained from MOG-i.v. treated mice were paradoxically immuno-stimulatory, as they induced a non specific Th1-type response both in vitro and in vivo. We conclude that blocking such APC activation would enhance the effectiveness of tolerance induction. PMID- 15748950 TI - Peroxisome proliferator-activated receptor-gamma agonists inhibit the activation of microglia and astrocytes: implications for multiple sclerosis. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma agonists, including thiazolidinediones (TZDs) and 15-deoxy-Delta(12,14) prostaglandin J(2) (15d PGJ(2)), have been shown to be effective in the treatment of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). This study aimed to compare the anti-inflammatory actions of three TZDs - rosiglitazone, pioglitazone, and ciglitazone - with those of 15d-PGJ(2) on stimulated mouse microglia and astrocytes. The results show that TZDs and 15d PGJ(2) are effective in inhibiting production of nitric oxide, the pro inflammatory cytokines TNF-alpha, IL-1beta, and IL-6, and the chemokine MCP-1 from microglia and astrocytes. However, 15d-PGJ(2) was a more potent suppressor of pro-inflammatory activity than the TZDs. These studies suggest that PPAR-gamma agonists modulate EAE, at least in part, by inhibiting the activation of microglia and astrocytes. The studies further suggest that PPAR-gamma agonists may be effective in the treatment of MS. PMID- 15748951 TI - Estrogen and brain inflammation: effects on microglial expression of MHC, costimulatory molecules and cytokines. AB - To model the effects of estrogen on adaptive immunity in the brain, we examined the effects of 17beta-estradiol on microglial parameters related to antigen presentation and T cell activation. Specifically, the effects of 17beta-estradiol on basal and LPS-induced surface staining of Class I and II MHC, as well as CD40, CD80, CD86, CD152, CD28, CD8, CD11b, Fas, FasL, and also ERalpha and ERbeta, were examined in N9 microglial cells. Additionally, the effects of 17beta-estradiol on basal and LPS-induced release of cytokines (TNF-alpha, IFN-gamma, IL-2, IL-4, and IL-10) were determined. Data indicate that estrogen increases IL-10 while decreasing TNFalpha and IFNgamma release from resting and LPS-stimulated N9 cells. Additionally, LPS-induced surface staining of MHC Class I, CD40, and CD86 was significantly attenuated by estrogen pretreatment. The basal percentage of cells positive for MHC Class I and II, CD40, and CD152, Fas, and FasL was significantly decreased by estrogen exposure. However, CD8, CD86, CD11b, and CD28 were unaffected by estrogen, and CD80 cell surface staining significantly increased following estrogen exposure. Taken together, these data indicate that estrogen can significantly decrease components of adaptive immunity in microglial cells, and highlight the multi-faceted regulatory effects of estrogen on microglial parameters related to antigen presentation and T cell interaction. PMID- 15748952 TI - Time course and role of the pineal gland in photoperiod control of innate immune cell functions in male Siberian hamsters. AB - The time course of select phagocyte and natural killer activities to short days was determined. In advance of testes regression, circulating granulocyte and monocyte cell numbers in hamsters decreased while lymphocyte numbers increased; phagocytosis and oxidative burst activity also decreased. To determine whether the pineal gland influences these innate immune cell functions, hamsters were exposed to constant light. Photoperiod control of testes weight and basal oxidative burst activity was abolished by treatment with constant light; other phagocyte activities and leukocyte proportions in circulation were not affected. The findings suggest that photoperiod and pineal gland function may regulate certain innate immune activities. PMID- 15748953 TI - Ocular inflammation alters swelling and membrane characteristics of rat Muller glial cells. AB - Ocular inflammation is a common cause of retinal edema that may involve swelling of Muller glial cells. In order to investigate whether endotoxin-induced ocular inflammation in rats alters the swelling and membrane characteristics of Muller cells, lipopolysaccharide (LPS; 0.5%) was intravitreally injected. At 3 and 7 days after treatment, hypotonic challenge induced swelling of Muller cell somata that was not observed in non-treated control eyes. Muller cells of LPS-treated eyes displayed a downregulation of inward K(+) currents and upregulation of A type K(+) currents that was associated with a decreased expression of Kir4.1 protein in retinal slices. The data suggest that ocular inflammation induces alterations of both the swelling characteristics and the K(+) channel expression of Muller cells. PMID- 15748954 TI - Disparate MHC class II haplotypes in myelin oligodendrocyte glycoprotein- and myelin basic protein-induced experimental autoimmune encephalomyelitis. AB - The major histocompatibility complex (MHC) regulates multiple sclerosis (MS) and its model experimental autoimmune encephalomyelitis (EAE). We created four new intra-MHC recombinant rat strains, between the MHC haplotypes RT1(n) (BN) and RT1(l) (LEW) on the LEW background, to define disease regulation and localization within the MHC. Immunization with recombinant myelin oligodendrocyte glycoprotein (a.a.1-125; MOG)/IFA induced EAE in strains expressing the MHC class II allele RT1.B(n), whereas strains expressing the RT1.B(l) were resistant. In myelin basic protein peptide (MBP(GP)63-88)/CFA-induced EAE, RT1.B(l) expressing strains were susceptible whereas strains expressing the RT1.B(n) were resistant. High levels of antigen-specific IFN-gamma secreting lymphoid cells and antigen-specific serum IgG antibodies were only recorded in rats with an MHC class II allele that permitted MOG- or MBP-EAE, respectively. Genetically, we localized the MHC regulation of the investigated EAE models to the central part of the MHC, containing the MHC class II (RT1.B/D) and the centromeric parts of the MHC class III. No influences were evident from the classical MHC class I (RT1.A), the telomeric parts of the MHC class III or the non-classical MHC class I (RT1.C/E/M) in contrast to previous reports. The MHC class II haplotype-specific regulation of EAE induced with two different CNS antigens demonstrates a strikingly specific MHC-association even within the same target organ. PMID- 15748955 TI - Changes in innate and acquired immune responses in mice with targeted deletion of the dopamine transporter gene. AB - The dopamine transporter (DAT) is responsible for the re-uptake of dopamine into presynaptic nerve terminals and thereby controls dopaminergic neurotransmission. Deletion of DAT results in a hyperdopaminergic phenotype and DAT(-/-) mice are characterized by pituitary hypoplasia, impaired maternal behavior, and increased locomotion. From earlier studies, we have evidence that the activity of the central dopaminergic system may play a role in determining immune reactivity and disease susceptibility. To further explore the functional relation between the dopaminergic system and the immune system, we investigated the activity of the immune system in DAT(-/-) mice. We show that in vitro, splenocytes from DAT(-/-) mice displayed reduced natural killer cell activity and reduced mitogen-induced cytokine responses. In contrast, LPS-induced cytokine production by macrophages was enhanced. In vivo, the cellular response to immunization with ovalbumine (OVA induced delayed type hypersensitivity response) was significantly reduced. Interestingly, the OVA-induced humoral response (anti-OVA IgG) was increased in DAT(-/-) mice compared to wild-type animals. Plasma levels of catecholamines and corticosterone did not differ significantly between DAT(-/-) and wild-type animals. In conclusion, we show in the present study that interfering with the dopaminergic system has major consequences for both the acquired and the innate immune response. PMID- 15748956 TI - Interferon-beta-1a induces increases in vascular cell adhesion molecule: implications for its mode of action in multiple sclerosis. AB - We investigated soluble vascular cell adhesion molecule-1 (sVCAM) levels and MRI lesions over 24 weeks in 15 Relapsing Remitting MS (RRMS) patients randomized prospectively to receive once-weekly (qw) IFN-beta-1a 30 mug intramuscularly (IM) (Group I, 8 patients) or three-times-weekly (tiw) IFN-beta-1a 44 mug subcutaneously (SC) (Group II, 7 patients). Both groups demonstrated a significant increase in sVCAM during treatment when compared to pre-treatment levels. Patients on IFN-beta-1a 44 mug SC tiw had a significant (p<0.0001) mean increase in sVCAM of 321.9 ng/ml which was significantly greater (p<0.0001) than with IFN-beta-1a 30 mug IM qw (68.6 ng/ml). There was a negative correlation between combined unique (CU) MRI lesions and sVCAM levels within the IFN-beta-1a 44 mug SC tiw group (slope=-0.00106, p=0.009). We postulate that the mode of action of IFN-beta therapy in MS may involve the induction of an increase in sVCAM. sVCAM could bind VLA-4 on T-cells and intercept their adhesion to the blood brain barrier (BBB). This mechanism is consistent with the observed clinical effect of IFN-beta in reducing MRI contrast enhancing lesions. PMID- 15748957 TI - Expression of a killer cell receptor-like gene in plastic regions of the central nervous system. AB - A property common to the immune system and the nervous system is regulation by a highly complex and adaptable network of cellular interactions. Major histocompatibility complex (MHC) class I molecules, which are ligands of antigen specific receptors on CD8 T cells and of inhibitory receptors on natural killer cells, have an important and surprising role in the control of activity-dependent neuronal plasticity in the central nervous system (CNS). While expression of MHC class I molecules in neurons has been reported, corresponding immune receptors have not been identified in the CNS. Here we show selective expression of a gene related to killer cell immunoglobulin-like receptor (KIR) genes in subregions of the mouse brain where synaptic plasticity and neurogenesis occur, including olfactory bulbs, rostral migratory stream and dentate gyrus of hippocampus. These results suggest new functions for KIR-like molecules in the CNS. PMID- 15748958 TI - Fas polymorphisms are associated with the presence of anti-ganglioside antibodies in Guillain-Barre syndrome. AB - Polymorphisms in genes involved in regulation of immune homeostasis may be a susceptibility factor in the induction of cross-reactive anti-ganglioside antibodies after infection in patients with Guillain-Barre syndrome (GBS). In this study we assessed whether polymorphisms in the promoter region of Fas and FasL and sFas and sFasL are related to GBS or its distinct clinical or serological subgroups. We show that the A(-670)G SNP in the promoter region of Fas and high levels of sFas are associated with the presence of anti-ganglioside antibodies, suggesting that Fas-FasL interaction is involved in the production of cross-reactive antibodies in GBS. PMID- 15748959 TI - In vivo detection of myelin proteins in cervical lymph nodes of MS patients using ultrasound-guided fine-needle aspiration cytology. AB - Cervical lymph nodes (CLN) have been described to be the first lymphoid draining site of the brain. In this study we used ultrasound guided fine needle aspiration cytology (USgFNAC) to obtain cells, in vivo, from non-enlarged CLN of multiple sclerosis (MS) patients and HCs (HC), and investigated whether myelin proteins could be detected. Macrophages containing myelin basic protein (MBP) and proteolipid protein (PLP) were found in CLN of all MS patients, whereas these could only be detected in a minority of HC. This novel approach allows investigation of the first draining site of the brain for in vivo analysis of the immune regulation of MS. PMID- 15748960 TI - Platelet-activating factor receptor gene polymorphism in Japanese patients with multiple sclerosis. AB - We evaluated the association of the platelet-activating factor receptor (PAFR) gene polymorphism (A224D) with the susceptibility and severity of multiple sclerosis (MS) in a Japanese population. DNA was collected from 162 Japanese patients with clinically definite 'conventional' MS (MS) and 245 healthy controls. The missense mutation A224D that impairs PAF-PAFR signaling was determined by polymerase chain reaction restriction fragment length polymorphism. The frequency of the AD/DD genotypes was significantly higher in MS patients (21.0%) than in healthy controls (13.5%) (p=0.045; odds ratio (OR), 1.71; 95% confidence interval (CI), 1.01-2.89). Moreover, the frequency of D allele in MS patients (11.7%) was also significantly higher than those in healthy controls (6.9%) (p=0.019; OR, 1.78; 95% CI, 1.10-2.89). These findings suggest that the PAFR gene missense mutation has a relation to the susceptibility for MS. PMID- 15748961 TI - The role of markers of bone remodeling in multiple myeloma. AB - Osteolytic bone disease is a frequent complication of multiple myeloma, resulting in skeletal complications that are a significant cause of morbidity and mortality. A characteristic feature of myeloma bone disease is that the lesions rarely heal and bone scans are often negative in myeloma patients who have extensive lytic lesions, offering very little in the follow-up of bone disease. X rays are also of limited value in monitoring bone destruction during anti-myeloma or anti-resorptive treatment. Biochemical markers of bone turnover, such as N- and C-terminal cross-linking telopeptide of type I collagen (NTX, CTX/ICTP, respectively), and newer ones such as the tartrate resistant acid phosphatase isoform 5b, provide information on bone dynamics that in turn may reflect disease activity in bone. Several studies have shown bone markers to be elevated in myeloma patients and reflect the extent of bone disease, while in some of them bone resorption markers correlate with survival. These markers may also be helpful in identifying those patients likely to respond to bisphosphonate treatment, and monitoring the effectiveness of bisphosphonate therapy in the management of myeloma bone disease. This review attempts to summarize the existing data for the role of markers of bone remodeling in assessing the extent of bone destruction in myeloma and monitoring bone turnover during specific anti myeloma treatment. We also discuss some novel markers that may be of particular interest in the near future. PMID- 15748962 TI - Management of acquired aplastic anaemia. AB - Outcome of patients with aplastic anaemia (AA), whether treated with allogeneic BMT or immunosuppressive therapy has steadily increased over the last three decades. However, there is a difference in quality of outcome between these two therapeutic modalities. There is no plateau for survival after ATG as patients are at later risk of transformation to myelodysplasia (MDS) or acute myeloid leukaemia (AML), paroxysmal nocturnal haemoglobinuria and relapse of their aplasia. In contrast, AA patients are not at risk of these later complications if they have undergone successful bone marrow transplantation. Long term survival after HLA identical sibling BMT is 80-90%, but GVHD and graft rejection remain to be addressed. The results of unrelated donor BMT for AA have shown considerable improvement over the last five years. Difficulties remain for those patients who fail immunosuppressive therapy and in whom BMT is not possible, since alternative immunosuppressive agents have so far proven to be somewhat disappointing. PMID- 15748963 TI - Kinetics of engraftment following allogeneic hematopoietic cell transplantation with reduced-intensity or nonmyeloablative conditioning. AB - Nonmyeloablative or reduced-intensity conditioning regimens have been used to condition elderly or ill patients with hematological malignancies for allogeneic hematopoietic cell transplantation (HCT). Initial mixed donor/host chimerism (i.e. the coexistence of hematopoietic cells of host and donor origin) has been observed in most patients after such transplants. Here, we describe both factors affecting engraftment kinetics in patients given a nonmyeloablative or a reduced intensity conditioning, and associations between peripheral blood cell subset chimerism levels and HCT outcomes. PMID- 15748966 TI - Management of infants with chronic lung disease of prematurity in various parts of the world. PMID- 15748964 TI - Follicular lymphoma: time for a re-think? AB - Follicular lymphoma (FL) is a malignancy of follicle centre B cells that have at least a partially follicular pattern, and is the commonest type of indolent Non Hodgkin's lymphoma. Except in the subset of patients with localized disease, FL should still be regarded as an incurable malignancy with a relentless relapsing/remitting course. However, the provocative new data covered by this review (including anti-CD20 antibody therapy, BCL-2, radioimmunotherapy, new chemotherapeutic agents and anti-idiotype vaccination), provides much cause for excitement and guarded optimism. Rituximab represents a novel treatment approach for a variety of disease settings, with a proven excellent efficacy and toxicity profile. Long-term data is required to establish whether its use translates into survival benefit. As the clinical activity of rituximab and other new therapeutic approaches becomes established, it will be important to determine how best to integrate these results into the standard care of patients with follicular lymphoma. PMID- 15748967 TI - Management of infants with chronic lung disease of prematurity in Australasia. AB - Chronic lung disease is common in extremely preterm infants born in Australasia. In 2002, 53% of surviving infants born before 28 weeks' gestation remained either oxygen-dependent or on other respiratory support at 36 weeks' postmenstrual age. In the first weeks of life oxygenation should be kept generally "lower", although what is the most appropriate level remains uncertain. During the mid-phase of the neonatal course, functional oxygen saturation levels around 90-95% probably confer the best benefit/risk balance. The most appropriate target saturation range for infants on home oxygen also remains uncertain. Definitive data to guide clinical practice is lacking regarding the use of postnatal corticosteroids, bronchodilators, and diuretics for either the treatment or prevention of chronic lung disease. Home oxygen programmes are effective in avoiding prolonged hospitalisation for infants with chronic lung disease, but require the coordination of a large, multidisciplinary team. PMID- 15748968 TI - Management of infants with chronic lung disease of prematurity in Chile. AB - Despite advances in the prevention and management of respiratory distress syndrome, chronic lung disease of prematurity (CLD) remains a major cause of morbidity and mortality in preterm babies in Chile. Its incidence varies from 10% to 60% in different regions of Chile. Since 1998, the management of CLD after discharge from neonatal unit follows national guidelines. Target oxygen saturation is 85% to 91% in the first 1 week of life, 91% to 94% from 1 to 2 weeks and over 95% after 44 weeks postconceptional age. National home oxygen program has improved outcome in infants with CLD. Other specific treatments are used with caution. Diuretics are used for pulmonary oedema. The adverse neurological outcome in infants treated with postnatal steroids restricts its use to infants who cannot be weaned from mechanical ventilation. Inhaled steroids and bronchodilators may reduce asthma-like symptoms in established CLD. Prevention of RSV infection in CLD babies is paramount. The preterm infant population has been maintained under surveillance nationally since 1998. PMID- 15748969 TI - Management of infants with chronic lung disease of prematurity in China. AB - With improved survival of very low birth weight infants in China over the last decade, chronic lung disease of prematurity (CLD) is only now becoming prevalent. As a result the management of CLD in China is only now beginning. In this paper, we describe the practice of managing these infants with as much evidence base as possible but often the management is based on other published papers in China and elsewhere and other people's personal experience. It appears that oxygen therapy is important to the survival of CLD infants but blood oxygen concentrations must be monitored closely in infants needing oxygen supplementation. We aim for a target range for oxygen saturation range of between 90%-95% to prevent retinopathy of prematurity. Although dexamethasone is effective in the treatment of CLD particularly extubation of preterm infants from mechanical ventilation, we restrict its use in severe infant due to their side effects. We have little experience of home oxygen and are only now setting up management protocols for oxygen use for CLD in both hospital and at home. We hope that the survival and outlook for these infants with CLD will improve over the next few years. PMID- 15748970 TI - Management of infants with bronchopulmonary dysplasia in Germany. AB - Although official guidelines for diagnosis and treatment of bronchopulmonary dysplasia (BPD) exist in Germany the practice in tertiary care neonatology centres differs considerably. There is no consensus about the appropriate level of oxygen saturation for infants at risk for BPD or infants with established BPD. Targeting oxygen saturation just below 90% in the first weeks and in the lower nineties thereafter seems to be a reasonable approach. Systemic corticosteroids must be used very restrictively because of adverse short- and long-term outcomes. Diuretics, inhaled corticosteroids, and bronchodilators may be used based on a stringent assessment of the individual response; their routine use cannot be recommended. Domiciliary oxygen is a therapy rarely prescribed in Germany although, if carefully planned and organised, it is safe and effective. Infants with home oxygen need a close follow-up by neonatologists and other specialists. Routine vaccination is recommended from the postnatal age of 3 months onwards. PMID- 15748971 TI - Management of infants with chronic lung disease of prematurity in the United Kingdom. AB - The management of chronic lung disease of prematurity (CLD) is challenging for the neonatologist. There are few well-powered randomised controlled trials to inform practice and longer-term outcomes of some interventions have only recently been identified (for example the possible association between the use of corticosteroids and neurodevelopmental abnormalities). As a result, many neonatologists rely on empirical management derived from the evidence available. We describe, in this article, our own approach to the management of CLD, and acknowledge that practice may vary between units within the United Kingdom. PMID- 15748972 TI - Management of infants with bronchopulmonary dysplasia in North America. AB - The in-hospital management of infants with BPD includes minimizing the duration of mechanical ventilation and avoiding the use of high inspired oxygen concentrations while maintaining adequate oxygenation. Fluid restriction, bronchodilators, and diuretic therapy can improve lung function and reduce the need for supplemental oxygen and high ventilator settings, but do not change the ultimate course of these infants. Corticosteroids also improve lung function and accelerate weaning from oxygen and mechanical ventilation, but their use during the first weeks of life is associated with worse neurological outcome. Adequate nutrition plays an important role in lung injury protection and recovery. Infants with severe BPD frequently develop pulmonary hypertension and may benefit from the use of pulmonary vasodilators. Outpatient management must be carefully planned and carried out by experienced multidisciplinary teams. Social and financial issues must be addressed with the family and caregivers. Home oxygen and mechanical ventilation therapy are used frequently after discharge and require specialized staff and equipment. Maintenance of oxygenation and proper nutritional support are critical aspects in the post-discharge management of these infants. Immunizations and RSV prevention are also important to prevent infections in these vulnerable immunocompromised patients. PMID- 15748973 TI - Maternal trait anxiety, depression and life event stress in pregnancy: relationships with infant temperament. AB - AIMS: To assess whether links exist between maternal trait anxiety (STAI), perceived life event (LE) stress and depression (Edinburgh scale) and infant temperament. STUDY DESIGN AND SUBJECTS: Women in the third trimester of pregnancy returned psychological self-report questionnaires; infant temperament was evaluated at 4 and 6 months by maternal and paternal report, while depression (concurrent Edinburgh scale) was also assessed at four and six months. As data were returned inconsistently at 4 and 6 months, we combined these two time points for simplicity of reporting and optimisation of numbers. RESULTS: Univariate logistic regressions on 970 subjects indicated that the pregnancy STAI (>40) scores were associated with 2.56- and 1.57-fold increases (maternal and paternal, respectively), in the odds of "difficult" infant temperament at 4 or 6 months. Concurrent Edinburgh scores (OR of 3.06 and 2.64 for maternal reports, respectively) were also predictive of infant temperament. Age, education, income, marital status, obstetric complications, infant gender and prematurity were not predictive of infant temperament. In stepwise multiple logistic regression analyses, the antenatal trait STAI (odds ratio 1.96) significantly predicted maternal reports of "difficult" temperament at 4 or 6 months independent of both antenatal and postnatal depression scores. There were similar trends for paternal reports of "difficult" temperament but these were not significant. Antenatal depression and perceived LE stress were not predictive of temperament. Finally, women (N=14) reporting domestic violence (DV) in pregnancy had highly significant increased Edinburgh and STAI scores. CONCLUSIONS: Maternal trait anxiety was predictive of "difficult" infant temperament, independent of "concurrent" depression and key sociodemographic and obstetric risk factors. These findings, while needing replication using objective measures of infant temperament, suggest that antenatal psychological interventions aimed at minimising anxiety may optimize infant temperament outcomes. There may be some benefit in shaping specific interventions to women reporting specific risk factors such as DV or past abuse. PMID- 15748974 TI - Urinary citrate in preterm and term babies. AB - OBJECTIVES: (1) To determine a normal range for urinary citrate for term babies. (2) To compare urinary citrate measured in ex preterm babies at term with this normal range. (3) To evaluate whether urinary citrate was related to presence of nephrocalcinosis (NC) and chronic Lung Disease (CLD) in these ex preterm babies. STUDY DESIGN: Urinary citrate was measured in 38 healthy term babies (mean birth weight 3.52 kg, mean gestation 41 weeks) at a mean postnatal age of 3 days (1-5 days) and in 53 ex preterm babies (<32 weeks gestation at birth) at term. These preterm babies were part of a larger study on NC in which two renal ultrasound scans were performed at 1 month and term. RESULTS: The normal range for urinary citrate in term babies was 0.025-2.97 (mean 1.03) mmol/l and citrate/creatinine ratio 0.0011-0.852 (mean 0.27). In the ex-preterm urinary citrate was not significantly different (mean 1.1 vs. 1.03, p=0.7232) but urine citrate /creatinine ratio was significantly higher (mean 1.27 vs. 0.27, p=0.0005). There was no significant difference in urinary citrate or ratios of citrate/creatinine and calcium/citrate in the 11 (20.7%) with NC or in the 17 (32%) babies with CLD. There was no significant relationship found between duration of TPN and urinary citrate measured at term. CONCLUSION: We have determined a normal range for urinary citrate in healthy term babies in the first week of life. The range was very wide. Ex preterm babies had similar values at term and there was no association between urinary citrate and NC or CLD. PMID- 15748975 TI - Duration of long-chain polyunsaturated fatty acids availability in the diet and visual acuity. AB - BACKGROUND: Little is known about the critical period during which the dietary supply of long-chain polyunsaturated fatty acids (LCPUFAs) may influence the maturation of visual cortical function in term infants. AIM: To define the relationship between duration of dietary LCPUFA supply and visual acuity at 52 weeks of age. STUDY DESIGN: Data from 243 infants who participated in four randomized clinical trials of LCPUFA supplementation of infant formula at a single research center were combined. The primary outcome was visual acuity at 52 weeks of age as measured by swept visual evoked potentials (sweep VEP). RESULTS: Longer duration of LCPUFA supply was associated with better mean acuity at 52 weeks of age (r=-0.878; p<0.001). The relationship between duration of dietary LCPUFA supply and sweep VEP acuity at 52 weeks was similar whether the LCPUFAs were provided via formula containing 0.36% DHA and 0.72% ARA or human milk. Duration of breast-feeding was associated with individual infants' sweep VEP acuity outcomes at 52 weeks (r=-0.286; p<0.005). The duration of LCPUFA supply during infancy has a similar relationship to sweep VEP acuity at 52 weeks in breastfed infants regardless of birth order. CONCLUSION: A continued benefit from a supply of LCPUFAs is apparent in infants through 52 weeks of age, suggesting that the brain may not have sufficient stores of LCPUFAs from an early postnatal supply to support the optimal maturation of the visual cortex. PMID- 15748976 TI - Twin-arginine-specific protein export in Escherichia coli. AB - In many prokaryotic organisms, secretory proteins harboring a twin-arginine consensus motif are exported in a fully folded conformation via the twin-arginine translocation (Tat) pathway. In Escherichia coli, Tat involves the three structurally and functionally different membrane proteins TatA, TatB, and TatC. Whereas TatC proteins function in the specific recognition of substrate, TatA might be the major pore-forming subunit. PMID- 15748977 TI - Detection of the cytolethal distending toxin locus cdtB among diarrheagenic Escherichia coli isolates from humans in Iran. AB - Cytolethal distending toxin (CDT) represents an emerging family of newly described bacterial products that are produced by a number of pathogens. Genes encoding this toxin were identified as a cluster of three adjacent genes, cdt-A, cdt-B and cdt-C. Five cdt genetic variants, designated cdt-I, cdt-II, cdt-III, cdt-IV and cdt-V, have been identified so far. To determine the presence of cdt genes among Escherichia coli isolates, a PCR assay was employed. Using multiplex primers designed for detection of E. coli cdt genes, PCR analysis indicated the presence of cdt genes in 37.5% of these isolates. While specific primers were located in the cdt-B locus for detection of cdt-I, cdt-II, cdt-III and cdt-IV, in multiplex PCR positive isolates indicated that a cdt-I-like gene was present in 45.3% of these isolates. However, in 52% of the isolates, the cdt-III-like gene could be detected. It should be mentioned that in 30.8% of these isolates, cdt-II and -III were detected simultaneously. In 2.6% of the isolates, the cdt-IV gene was present. The cnf-1 gene was detected in 29.4% of strains carrying the cdt-I gene; however, the cnf-2 gene was detected only in 23.1% of the cdt-III-like strains. Furthermore, the data obtained by PCR analysis indicated the presence of cdt-like genes among our E. coli isolates, although in the CHO cell assay, all isolates showed a cytopathic effect characteristic of CDT. PMID- 15748978 TI - Structural and biochemical analysis of the asc operon encoding 6-phospho-beta glucosidase in Pectobacterium carotovorum subsp. carotovorum LY34. AB - An asc operon of Pectobacterium carotovorum subsp. carotovorum LY34 (Pcc LY34) was isolated from a genomic library in a screen for beta-glucosidase activities. Sequence analysis of the 5618-bp cloned DNA fragment (accession number AY622309) showed three open reading frames (ascG, ascF, and ascB) that are predicted to encode 375, 486, and 476 amino acid proteins, respectively. The AscG ORF shared a high similarity with the Escherichia coli AscG repressor. The AscF ORF shared 81% identity with the E. coli AscF PTS enzyme II(asc), while the AscB ORF was highly similar to 6-phospho-beta-glucosidases and is a member of the glycosyl hydrolase family 1. The purified AscB enzyme hydrolyzed salicin, arbutin, pNPG, and MUG. It exhibited maximal activity at pH 7.0 and 40 degrees C, and its activity was enhanced in the presence of Mg(2+) and Ca(2+). The molecular weight of the enzyme was estimated to be 53 000 Da by SDS-PAGE. Two conserved glutamate residues (Glu(182) and Glu(374)) were shown to be important for AscB activity. PMID- 15748979 TI - Safety evaluation of Lactobacillus delbrueckii subsp. lactis UO 004, a probiotic bacterium. AB - Lactobacillus delbrueckii subsp. lactis UO 004 was evaluated for its use as a potential probiotic from a safety point of view. The strain did not exhibit mucinolytic or other enzymatic activities that might be detrimental, such as those involving glycosidases (beta-D-glucosaminidase or alpha-D-galactosidase) or arylamidases (factor Xa and quimotrypsin-like activities), frequently present in Lactobacillus strains isolated from patients with endocarditis, although it was able to express protein Ca and kallikrein-like activities. On the other hand, the presence of the strain did not interfere with the growth of certain species of normal intestinal microbiota, such as Enterococcus fecalis, Escherichia coli, Bifidobacterium bifidum or Bacteroides fragilis. Moreover, the potential probiotic strain UO 004 is sensitive to antibiotics with transmissible resistance mechanisms in Lactobacillus such as chloramphenicol, erythromycin, tetracycline and vancomycin. In addition, strain L. delbrueckii UO 004 was not able to translocate towards the intestinal barrier of mice or produce changes in their activity or general health status. PMID- 15748980 TI - Sequence heterogeneity in the lacSZ operon of Streptococcus thermophilus and its use in PCR systems for strain differentiation. AB - Sequences of the lacSZ operon of 29 Streptococcus thermophilus strains from different dairy products were determined. Differences in sequence among the strains were detected within LacS more often than in the LacZ gene. The sequences were aligned and compared and it was possible to gather the strains into three groups of similarity on the basis of the LacS gene sequence. The dairy environment of origin did not seem to be related to the lacSZ operon sequence and thus to the similarity shown. Nucleotide variability was investigated and a total of 139 nucleotide changes were found in the LacS gene while 40 nucleotide changes were found in the sequences of the LacZ gene. Moreover, the influence of the nucleotide changes on the amino acid sequence of the LacS transporter and of the beta-galactosidase enzyme were discussed. Sequence variability within the region upstream from the LacS gene was used to develop group-specific PCR systems capable of distinguishing S. thermophilus at the strain level. A strain-specific primer set was designed allowing the specific detection of 11 out of 29 strains of S. thermophilus. Moreover, LacS-PCR-SSCP analysis of the 29 strains provided 2 different profiles, whereas 4 strain-specific profiles were detected by LacS-PCR DGGE, indicating the potential to use these techniques for profiling and monitoring population of strains of S. thermophilus in food products. The results are discussed with reference to the potential of these PCR methods for ascertaining strain dominance and starter fitness in dairy processes. PMID- 15748981 TI - Glutamine amidotransferase activity of NAD+ synthetase from Mycobacterium tuberculosis depends on an amino-terminal nitrilase domain. AB - NAD(+) synthetase (NadE; E.C. 6.3.5.1) from Mycobacterium tuberculosis utilizes both glutamine and ammonia to catalyze NAD(+) production, in contrast to the corresponding NH(3)-dependent enzymes from other prokaryotes. Here we report the site-directed mutagenesis of amino acids located in the N-terminal domain and predicted to be essential for glutamine hydrolysis. The residues forming the putative catalytic triad (Cys176, Glu52 and Lys121) were replaced by alanine; the mutated enzymes were expressed in the Escherichia coli Origami (DE3) strain and purified. The three mutants completely lost their glutamine-dependent activity, clearly indicating that Cys176, Glu52 and Lys121 are crucial for this activity. In contrast, the C176A and E52A variants, respectively, retained 90 and 30% of the original NH(3)-dependent specific activity, while the K121A mutant lost this activity. The results show that glutamine-amidotransferase activity is mediated by an N-terminal domain belonging to the superfamily of nitrilases. This domain, a new type of glutamine amide transfer (GAT) domain, is the first to be characterized in bacterial NAD(+) synthetases. PMID- 15748982 TI - The role of isocitrate lyase and the glyoxylate cycle in Escherichia coli growing under glucose limitation. AB - Escherichia coli changes its metabolism in response to environmental circumstances, and metabolic adaptations are evident in hungry bacteria growing slowly in glucose-limited chemostats. The role of isocitrate lyase (AceA) was examined in E. coli growing under glucose limitation. AceA activity was elevated in a strain-dependent manner in the commonly used E. coli K-12 laboratory strains MG1655 and MC4100, but an aceA disruption surprisingly increased fitness under glucose limitation in both strains. However, in bacteria adapted to limiting glucose in long-term chemostats, mutations outside aceA changed its role from a negative to a positive influence. These results suggest that a recently proposed pathway of central metabolism involving the glyoxylate cycle enzymes is redundant in wild-type bacteria, but may take on a beneficial role after context adaptation. Interestingly, the aceA gene sequence did not alter during prolonged selection, so mutations in unidentified genes changed the metabolic context of unaltered AceA from a negative to a positive influence in bacteria highly adapted to limiting glucose. PMID- 15748983 TI - Nitrogen nutrition and isotopic discrimination in tropical ectomycorrhizal fungi. AB - It is known that many ectomycorrhizal (EcM) fungi are able to utilise complex organic sources of nitrogen. Two hypotheses were tested using isolates of tropical EcM fungi grown in vitro: (i) EcM fungi isolated from mineral soils of tropical rain forests are less able to utilise organic sources of nitrogen than mineral sources; and (ii) nitrogen isotope discrimination patterns follow those of the nitrogen source utilised. Pisolithus albus and Tomentella sp. represented tropical EcM fungi and they were grown along with Thelephora terrestris. All three species were able to utilise bovine serum albumen as a nitrogen source and P. albus produced the greatest biomass on this source of nitrogen. Nitrate was generally less well utilised than ammonium although all three species were able to grow on this nitrogen source. The nitrogen source which led to the greatest biomass also led to the highest fungal nitrogen concentration in P. albus and Tomentella sp., but not T. terrestris. All three species discriminated against (15)N when grown on BSA and NO(3) but there were interspecific differences in isotope discrimination when grown on NH(4). From a limited number of isolates, it was found that EcM fungi from tropical soils utilise protein nitrogen as well as mineral nitrogen and there can be considerable nitrogen isotope discrimination during the utilisation of all these nitrogen sources. PMID- 15748984 TI - Characterization of Saccharomyces cerevisiae natural populations for pseudohyphal growth and colony morphology. AB - In this work we have analyzed the colony and cellular morphologies of natural populations of Saccharomyces cerevisiae strains in response to different environmental stimuli. Among one thousand strains grown on YPD medium, 2.5% exhibited a rough (R) colony phenotype versus a smooth (S) phenotype. When grown on the ammonium-deficient medium SLAD, 56% of the strains showed a filamentous phenotype, often associated (43.8%) with an invasive phenotype, while 4.7% of the strains exhibited only an invasive phenotype. The rough phenotype on YPD was always associated with the filamentous phenotype on SLAD. A subset of 52 strains was further characterized for the growth phenotype under different stimuli (nitrogen deprivation, addition of alcohols, growth on proline as sole nitrogen source). On 27 strains, genetic analysis of the spore products was also performed. The entire set of data showed a wide distribution of dimorphism in the yeast population and great variability with respect to the dimorphic switch capability. Some strains grew with peculiar colony morphologies under different environmental stimuli and some showed colony morphology variations. Ecological implications of the wide spreading of dimorphic behavior and the occurrence of peculiar colony morphologies in natural yeasts are discussed. PMID- 15748986 TI - Transcriptional response of Rickettsia conorii exposed to temperature variation and stress starvation. AB - Rickettsia conorii is an obligate intracellular bacterium transmitted to humans by Rhipicephalus sanguineus ticks. The success of this microorganism at surviving in nature implicates the ability to efficiently adapt to different environments, including the arthropod vector and the mammalian host. Numerous bacterial species possess a highly evolved system for stress adaptation. This so-called stringent response is mediated by guanosine 3',5'-bispyrophosphate and guanosine 3' diphosphate 5'-triphosphate which are under spoT control in some Gram-negative bacteria. Interestingly, annotation of the R. conorii genome evidenced 5 spoT paralogs. We hypothesized that these spoT genes play a role in adaptation to environmental changes specifically encountered by rickettsiae during their different life cycles. Transcription of the spoT paralogs was examined by RT-PCR from infected Vero cells maintained in rich or deficient culture media, from infected C6/36 insect cells cultured at various temperatures and from infected ticks. Our results demonstrated that the 5 spoT genes can be transcribed. SpoT1 (RC0374) is only transcribed upon stringent response. Transcription of spoT3 (RC0888) was never observed in arthropod cells or ticks, but was specific to R. conorii RNA extracted from infected Vero cells. These results indicate that rickettsial spoT paralogs are independently transcribed, depending on the different infected hosts and the adaptive capacity of the pathogen. Bioinformatics analysis of these possibly encoded proteins is also reported. PMID- 15748985 TI - T-RFLP analysis of bacterial communities in cyclodextrin-amended bioreactors developed for biodegradation of polychlorinated biphenyls. AB - In this study, T-RFLP analysis was used to determine the structure and spatial distribution of the indigenous bacterial community of an actual-site PCB contaminated soil treated in aerobic packed-bed loop reactors (PBLRs) in the absence or in the presence of a mixture of randomly methylated beta-cyclodextrins (RAMEB) at 0.5 or 1% w/w. RAMEB was found to significantly enhance the aerobic bioremediation of soil with effects that increased proportionally with the concentration at which it was applied. At the end of treatment (180 days), T-RFLP analysis of the soil samples collected from the top and bottom regions of the PBLRs showed a series of 50 single T-RFs. Remarkably, the number of T-RFs was significantly lower (13-22) in samples collected from different sections of the RAMEB-amended bioreactors with respect to equivalent samples collected from the RAMEB-free reactor. Cluster analysis based on the presence or the absence of T RFs peaks revealed high similarity, inside each reactor, between the top and bottom parts of its soil bed. Soil samples collected at the top and bottom regions of the two bioreactors amended with RAMEB, clustered together while the equivalent samples of the bioreactor without RAMEB formed a separate cluster which was distantly related to the soil samples obtained from the parallel amended bioreactor. Notably, T-RFLP analyses combined with extensive sequencing of 16S rDNA allowed us to tentatively allocate a series of bacterial species corresponding to specific peaks of the T-RFLP profiles and to determine their phylogenetic affiliation. PMID- 15748987 TI - Cloning and characterization of the hsp 18.55 gene, a new member of the small heat shock gene family isolated from wine Lactobacillus plantarum. AB - Using a molecular approach based on PCR, RT-PCR and northern blot analysis, a new member of the small heat shock family of wine, Lactobacillus plantarum, was cloned and characterized. The protein sequence deduced from the isolated gene had a calculated molecular mass of 18.548 kDa and was therefore named HSP 18.55. The gene codes for a protein homologous to the previously characterized HSP 19.3 and HSP 18.5 and is co-transcribed with an upstream gene of unknown function. Analysis of the 5' flanking region of the hsp 18.55 gene revealed the presence of putative cis elements able to bind alternative sigma factor sigma(B). Based on its structure, the gene was classified as belonging to class II of the heat shock genes according to Bacillus subtilis nomenclature for shock-responsive genes. Expression of the newly identified small heat shock gene, analyzed by RT-PCR and northern blot analysis, was induced by a wide range of abiotic stresses including heat, cold and ethanol, suggesting that the small family of heat shock genes is probably involved in the general stress response in wine L. plantarum. Moreover, the expression of hsp 18.5, hsp 18.55 and hsp 19.3 genes, analyzed over a complete culture cycle, revealed that early growing cells contained substantial amounts of hsp 18.5, hsp 18.55 and hsp 19.3 mRNAs, which rapidly declined upon entry into stationary phase. PMID- 15748988 TI - Unique stress response to the lactoperoxidase-thiocyanate enzyme system in Escherichia coli. AB - Using a differential fluorescence induction approach, we screened a promoter trap library constructed in a vector with a promoterless gfp gene for Escherichia coli MG1655 promoters that are induced upon challenge with the antimicrobial lactoperoxidase-thiocyanate enzyme system. None of the thirteen identified lactoperoxidase-inducible open reading frames was inducible by H(2)O(2) or by the superoxide generator plumbagin. However, analysis of specific promoters of known stress genes showed some of these, including recA, dnaK and sodA, to be inducible by the lactoperoxidase-thiocyanate enzyme system. The results show that the lactoperoxidase-thiocyanate enzyme system elicits a distinct stress response different from but partly overlapping other oxidative stress responses. Several of the induced genes or pathways may be involved in bacterial defense against the toxic effects of the lactoperoxidase-thiocyanate enzyme system. PMID- 15748989 TI - SulA-dependent hypersensitivity to high pressure and hyperfilamentation after high-pressure treatment of Escherichia coli lon mutants. AB - High-pressure treatment (>100 MPa) is known to induce several heat shock proteins as well as an SOS response in Escherichia coli. In the current work, we have investigated properties with respect to high-pressure treatment of mutants deficient in Lon, a pressure-induced ATP-dependent protease that belongs to the heat shock regulon but that also has a link to the SOS regulon. We report that lon mutants show increased pressure sensitivity and exhibit hyperfilamentation during growth after high-pressure treatment. Both phenotypes could be entirely attributed to the action of the SOS protein SulA, a potent inhibitor of the cell division ring protein FtsZ and a specific target of the Lon protease, since they were suppressed by knock-out of SulA. Introduction of the lexA1 allele, which effectively blocks the entire SOS response, also suppressed the high pressure hypersensitivity of lon mutants, but not their UV hypersensitivity. These results indicate the existence of a SulA-dependent pathway of high-pressure-induced cell filamentation, and suggest involvement of the SOS response, and particularly of SulA, in high-pressure-mediated cell death in E. coli strains which are compromised in Lon function. PMID- 15748990 TI - Adhesion of bacteria to resected human colonic tissue: quantitative analysis of bacterial adhesion and viability. AB - Adhesion to the intestinal mucosa is considered to be one of the main selection criteria of lactic acid bacteria for probiotic use. Adhesive probiotics are, for example, considered to provide better antagonism against pathogenic bacteria when compared to non-adhesive strains. Here a new model is described for studying adhesion and interaction of probiotic and pathogenic bacteria in the intestinal mucus in which the intestinal microbiota is present. The model is based on the use of human intestinal tissue, fluorescent-tagged bacteria and confocal laser scanning microscopy (CLSM) in adhesion measurements as well as human intestinal mucus and bioluminescent-tagged bacteria in viability measurements. Use of CLSM enabled, for the first time, real-time three-dimensional observations of live probiotic bacteria in their natural environment, the intestinal mucosa. When the real-time measurement of bacterial adhesion was combined with the real-time sensitive measurement of bacterial viability, it could be studied whether or not the adherent pathogens were alive. The model was used to study the interaction between Lactobacillus rhamnosus GG and Salmonella enterica serovar Typhimurium. We show that L. rhamnosus GG did not affect the adhesion or the viability of S. enterica serovar Typhimurium. Instead S. enterica serovar Typhimurium was shown to decrease the adhesion of L. rhamnosus GG in displacement assays. Moreover, the method is suitable for studies in which the interaction of two or more bacteria is examined in an environment in which other bacteria are present. PMID- 15748991 TI - A new family of mobilizable suicide plasmids based on broad host range R388 plasmid (IncW) and RP4 plasmid (IncPalpha) conjugative machineries and their cognate Escherichia coli host strains. AB - We describe the construction of the pSW family of conditionally replicating plasmids which are based on the IncX oriV origin (oriV(R6Kgamma)) of replication that is dependent on the pir-encoded protein. We constructed several Escherichia coli derivatives expressing pir from different chromosomal loci, and the pir gene could be transduced by phage P1 to any E. coli strain. These chromosomal constructions generate dapA and thyA knockouts, which lead to diaminopimelate or thymidine auxotrophies, respectively, and they serve to provide absolute counterselection even in rich media. These strains can be easily counterselected if used in plasmid transfer experiments into markerless recipients, and they have been demonstrated to work efficiently in E. coli xVibrio or E. coli xBartonella matings. We constructed different pSW plasmids carrying either the oriT(RP4) or the oriT(R388), and we demonstrated that these derivatives can be efficiently transferred using RP4 and R388 conjugation machineries, respectively. We also constructed two plasmids expressing the R388 conjugation machinery, but lacking the oriT(R388). We demonstrated that these plasmids enabled efficient and exclusive transfer of a pSW-oriT(R388) derivative from E. coli to V. cholerae, and we offer an alternative to the popular RP4-based delivery system. PMID- 15748992 TI - Lactobacillus casei DN-114 001 inhibits the increase in paracellular permeability of enteropathogenic Escherichia coli-infected T84 cells. AB - Probiotics are living microorganisms which, when ingested in adequate amounts, exert health benefits toward the host. For instance, probiotics might act through reinforcement of the intestinal epithelial barrier function. The goal of the present study was to determine whether Lactobacillus casei DN-114 001 could abrogate the increase in paracellular permeability induced by enteropathogenic Escherichia coli. We used the human colon T84 cell line infected with a wild-type enteropathogenic E. coli (strain E2348/69). Paracellular permeability was followed by monitoring transepithelial electrical resistance variations and by observing zonula occludens-1 distribution. Two infection procedures were used: co incubation (the pathogenic and probiotic strains were simultaneously incubated with T84 cells) and post-infection (the probiotic was added in the presence of pathogenic bacteria 3 h after the beginning of the infection). We also investigated the effect of L. casei on enteropathogenic E. coli adhesion. L. casei DN-114 001 inhibited, in a dose-dependent-manner, the decrease in enteropathogenic E. coli-induced transepithelial electrical resistance and zonula occludens-1 redistribution using two different infection procedures. However, L. casei did not inhibit pathogenic strain adhesion. L. casei DN-114 001 inhibited the increase in EPEC-induced paracellular permeability. This property could partially explain the previously observed health benefits of this probiotic for human natural defenses, such as those associated with prevention of diarrhea. PMID- 15748993 TI - Pathology of Anticarsia gemmatalis larvae infected by two recombinant A. gemmatalis multicapsid nucleopolyhedroviruses. AB - Light and stereomicroscopy examinations of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV)-infected insects were performed in order to follow infection in its host, A. gemmatalis. Fourth-instar A. gemmatalis larvae were infected by administration of occluded virus (polyhedra) from two recombinant AgMNPV viruses (vAgEGTDelta-lacZ or vAgGalA2) directly into the larvae foregut. The recombinant virus vAgEGTDelta-lacZ has the beta-galactosidase gene (lac-Z) of Escherichia coli under the control of a constitutive promoter (hsp70 from Drosophila melanogaster). The vAgGalA2 virus has the reporter gene lac-Z under the control of the AgMNPV very late polyhedrin gene promoter. At different times post-infection (p.i.) the infected larvae were dissected, fixed, and the product of the expression of the lac-Z gene detected by incubating the insects in a buffer containing X-gal. This allowed us to follow the infection through the blue cells (due to the degradation of X-gal by the enzyme Lac-Z). Insect larvae inoculated with polyhedra from the recombinant viruses showed midgut cells to be infected first, followed by tracheal cells, hemolymph, fat body, Malpighian tubules and brain cells. The infection was similar for the two recombinant viruses, with blue cells appearing earlier in insects infected with the vAgEGTDelta-lacZ virus when compared to the vAgGalA2 virus. PMID- 15748994 TI - Topological analysis of integral membrane constituents of prokaryotic ABC efflux systems. AB - The ATP binding cassette (ABC) superfamily consists of dozens of families of transport systems, each of which catalyzes uptake or efflux of a specific type of molecule using ATP hydrolysis to energize transport. While all of the ATP hydrolyzing subunits in the superfamily are homologous, a monophyletic origin of the integral membrane constituents is not established. We have identified a subset of these transmembrane proteins that have a basal unit of four transmembrane alpha-helical segments (TMSs) with a large extracytoplasmic domain between TMSs 1 and 2. These homologues were found to exhibit 4, 8 or 10 putative TMSs per polypeptide chain. The two larger topological types exhibit a 4 TMS repeat element resulting from an internal gene duplication event, and the 10 TMS proteins display an extra two putative TMSs between the two repeat units. Rare intragenic deletions in these homologues gave rise to truncated proteins lacking the extracytoplasmic domain, and some phylogenetic clusters of the 4 TMS membrane proteins (but not the 8 or 10 TMS proteins) are fused N-terminal (never C terminal) to ATP hydrolyzing domains. Bioinformatic analyses lead to the suggestion that in the larger homologues, the second repeat units are more important for function than the first repeat units. Operon analyses suggest that the 4 TMS proteins form heterodimeric complexes while the 8 and 10 TMS proteins incorporate the equivalent of these complexes into single integral membrane polypeptide chains. Different gene compositions of the operons encoding the 4 versus 8 and 10 TMS homologues suggest that these two structural types of transporters act on different types of substrates and serve dissimilar functions. Significant sequence similarity between the integral membrane constituents of the ABC efflux pumps analyzed here and those of other ABC transporters could not be detected. These studies define the evolutionary pathway taken for the appearance of a subset of ABC transmembrane transport proteins and provide clues regarding their mechanistic and functional characteristics. PMID- 15748995 TI - Genome-wide identification and mapping of variable sequences in the genomes of Burkholderia mallei and Burkholderia pseudomallei. AB - Burkholderia mallei and Burkholderia pseudomallei, closely related Gram-negative bacteria, are the causative agents of such serious infectious diseases of humans and animals as glanders and melioidosis, respectively. Despite numerous studies of these pathogens, the detailed mechanisms of their pathogenesis is still poorly understood. One of the serious obstacles to revealing factors responsible for pathogenicity lies in the considerable natural variability of B. pseudomallei and B. mallei, which is also a challenge to development of rapid and efficient diagnostic tools facilitating unambiguous identification of the infectious agents. To gain a deeper insight into B. mallei and B. pseudomallei interspecies divergence and intraspecies polymorphism, we compared the genomes of B. mallei C 5 and B. pseudomallei C-141 strains using a subtractive hybridization technique. A library of DNA fragments specific for B. mallei C-5 and absent from B. pseudomallei C-141 was obtained and analyzed. Some of the differential sequences detected were also not found in the recently sequenced genome of B. pseudomallei K96243. However, a multitude of B. mallei C-5 sequences absent from the B. pseudomallei C-141 genome were detected in the genome of B. pseudomallei K96243. On the other hand, some sequences identified as constituents of the B. mallei C-5 genome were not found in the genome of B. mallei ATCC 23344. Some of the differential DNA fragments displayed similarity to different mobile elements that have not yet been described for B. mallei, whereas the others matched fragments of various prophages, or, when translated into protein sequences, components of active transport systems and different enzymes. A substantial proportion of the differential clones had no database matches either at the nucleotide or amino acid sequence level. The results suggest great genome-wide intra- and interspecies variability of B. mallei and B. pseudomallei. The differences identified may be useful as molecular signatures for identification of B. mallei strains. PMID- 15748996 TI - From the editor's desk. PMID- 15748997 TI - Versican protects cells from oxidative stress-induced apoptosis. AB - Oxidant injury plays a critical role in the degenerative changes that are characterized by a decline in parenchymal cell numbers and viability, and occur with aging and in the etiology of many diseases. The extracellular proteoglycan versican is widely distributed in the extracellular matrix surrounding the cells. This study examines whether versican plays a role in protecting cells from free radical-induced apoptosis. Stable expression of versican or its C-terminal domain significantly decreased H(2)O(2)-induced cellular apoptosis. Cells in adherent monolayer were more resistant to H(2)O(2)-induced apoptosis than cells cultured in suspension. While vigorous trypsinization caused integrin cleavage and rendered the cells more susceptible to H(2)O(2)-induced damages, expression of versican or its C-terminal domain enhanced cell attachment and expression of beta1 integrin and fibronectin. Enhanced cell-matrix interaction by addition of manganese (MnCl(2)) to cultures also significantly diminished H(2)O(2)-induced apoptosis. The results suggest that versican plays an important role in reducing oxidant injury through an enhancement of cell-matrix interaction. PMID- 15748998 TI - Heparan sulphate interacts with tropoelastin, with some tropoelastin peptides and is present in human dermis elastic fibers. AB - A number of reports point to the presence of proteoglycans and/or glycosaminoglycans within elastic fibers in normal and in pathological conditions. We present data that heparan sulphate (HS)-containing proteoglycans are associated with normal elastic fibers in human dermis and that isolated HS chains interact in vitro with recombinant tropoelastin and with peptides encoded by distinct exons of the human tropoelastin gene (EDPs). By immunocytochemistry, HS chains were identified as associated with the amorphous elastin component in the human dermis and remained associated with the residual elastin in the partially degenerated fibers of old subjects. HS appeared particularly concentrated in the mineralization front of elastic fibers in the dermis of patients affected by pseudoxanthoma elasticum (PXE). In in vitro experiments, HS induced substantial changes in the coacervation temperature and in the aggregation properties of recombinant tropoelastin and of synthetic peptides (EDPs) corresponding to sequences encoded by exons 18, 20, 24 and 30 of the human tropoelastin gene. In particular, HS modified the coacervation temperature and favoured the aggregation into ordered structures of tropoelastin molecules and of EDPs 18, 20 and 24, but not of EDP30. These data strongly indicate that HS elastin interactions may play a role in tissue elastin fibrogenesis as well as modulating elastin stability with time and in diseases. PMID- 15748999 TI - The antiangiogenic effect of thrombospondin-2 is mediated by CD36 and modulated by histidine-rich glycoprotein. AB - Thrombospondins-1 and -2 (TSP-1, TSP-2) are matricellular glycoproteins with potent antiangiogenic activity. We have previously shown that the antiangiogenic activity of TSP-1 is mediated by the interaction of the type I repeats (TSR) with the receptor CD36, although other domains of TSP-1 have also been implicated. We now show that the antiangiogenic activity of TSP-2, which contains three TSRs but, unlike TSP-1, lacks the capacity to activate TGF-beta, is similarly dependent on CD36. Using the corneal pocket assay we found that TSP-2 did not inhibit bFGF-induced angiogenesis in CD36 null mice. We then demonstrated that (125)[I]-TSP-2 bound to murine macrophages and that binding was diminished by 70% by anti-CD36 antibody or by using cells from CD36 null animals. Solid-phase binding studies revealed that (125)[I]-TSP-2 bound to CD36/glutathione-S transferase (GST) fusion proteins encoding the region spanning amino acids 93 120, but not amino acids 298-439. This 93-120 amino acid region, previously identified as the TSP-1 binding site, is homologous to domains on other TSP binding proteins, such as LIMP-2 and histidine-rich glycoprotein (HRGP). Finally, we showed with an immunoabsorbent binding assay that TSP-2 bound HRGP with high affinity and that HRGP blocked the antiangiogenic activity of TSP-2, acting like a "decoy" receptor. These data suggest that modulation of the TSR/CD36 system may play an important role in the regulation of the angiogenic "switch," and may provide a target for therapeutic interventions. PMID- 15749000 TI - Cholesterol secretion and homeostasis in chondrocytes: a liver X receptor and retinoid X receptor heterodimer mediates apolipoprotein A1 expression. AB - Cholesterol is required for chondrocyte differentiation and bone formation. Apolipoprotein A1 (apoA-1) plays a major role in lipoprotein clearance and cholesterol redistribution. We report here that apoA-1 is expressed during chondrocyte differentiation in vitro and in vivo. In differentiating chondrocytes, the expression of the liver X receptor (LXR) is modulated and its expression correlates to the expression of apoA-1. The expression of other LXR target genes related to cholesterol homeostasis such as ABCA1 cholesterol transporter and sterol regulatory element-binding protein 1 (SREBP1) is similarly regulated. Small molecule ligands activating either LXR or retinoid X receptor (RXR) lead to a dramatic increase in apoA-1 mRNA and protein expression in cultured chondrocytes. These ligands strongly induce ABCA1 cholesterol transporter expression and effectively mediate cholesterol efflux from hypertrophic chondrocytes. In addition, we report that, in the same cells, the ligands down modulate Serum Amyloid A expression induced by bacterial lipopolysaccharide. Our studies provide evidence that LXR/RXR mediate a fine regulation of cholesterol homeostasis in differentiating chondrocytes. PMID- 15749001 TI - Quantitative analysis and comparative regional investigation of the extracellular matrix of the porcine temporomandibular joint disc. AB - Characterization of the extracellular matrix of the temporomandibular joint (TMJ) disc is crucial to advancing efforts in tissue engineering the disc. However, the current literature is incomplete and often contradictory in its attempts to describe the nature of the TMJ disc matrix. The aim of this study was to identify the variation of key matrix components along the three axes of the porcine disc using ELISAs to quantify these matrix components, immunohistochemistry to identify their regional distribution, and SEM to examine collagen fiber diameter and orientation. The overall GAG content of the TMJ disc (including the dermatan sulfate proteoglycans) was 5.3+/-1.2% of the dry weight. Chondroitin sulfate, which comprised 74% of this total GAG content, was 4.4, 8.2, and 164 times more abundant than dermatan sulfate proteoglycan, keratan sulfate, and hyaluronic acid, respectively. In general, these GAGs were most concentrated in the intermediate zone of the TMJ disc, appearing in dense clusters, and least concentrated in the posterior band. Additionally, chondroitin sulfate was more abundant medially than laterally. Collagen II was discovered in trace amounts, with higher relative amounts in the intermediate zone. Collagen fibers were observed to run primarily in a ring-like fashion around the periphery of the disc and anteroposteriorly through the intermediate zone, with a mean fiber diameter of 18+/-9 mum. Characterization studies of the TMJ disc, including prior biomechanical and cell studies along with the current study of the extracellular matrix, collectively reveal a distinct character of the intermediate zone of the disc compared to its anterior and posterior bands. PMID- 15749002 TI - High-level production of human collagen prolyl 4-hydroxylase in Escherichia coli. AB - The collagen prolyl 4-hydroxylases (C-P4Hs), enzymes residing within the lumen of the endoplasmic reticulum, play a central role in the synthesis of all collagens. The vertebrate enzymes are alpha(2)beta(2) tetramers in which the two catalytic sites are located in the alpha subunits, and protein disulfide isomerase serves as the beta subunit. All attempts to assemble an active C-P4H tetramer from its subunits in in vitro cell-free systems have been unsuccessful, but assembly of a recombinant enzyme has been reported in several cell types by coexpression of the two types of subunit. An active type I C-P4H tetramer was obtained here by periplasmic expression in Escherichia coli strains BL21 and RB791. Further optimization for production by stepwise regulated coexpression of its subunits in the cytoplasm of a thioredoxin reductase and glutathione reductase mutant E. coli strain resulted in large amounts of human type I C-P4H tetramer. The specific activity of the C-P4H tetramer purified from the cytoplasmic expression was within the range of values reported for human type I C-P4H isolated as a nonrecombinant enzyme or produced in the endoplasmic reticulum of insect cells, but the expression level, about 25 mg/l in a fermenter, is about 5-10 times that obtained in insect cells. The enzyme expressed in E. coli differed from those present in vivo and those produced in other hosts in that it lacked the N glycosylation of its alpha subunits, which may be advantageous in crystallization experiments. PMID- 15749003 TI - Identification and characterization of a second fibronectin gene in zebrafish. AB - Fibronectin (FN) is a highly conserved extracellular matrix protein that plays crucial roles in vertebrate embryogenesis. Previously, it was reported that zebrafish possess a single FN gene (fn1a). Here we report the presence of a second zebrafish FN gene (fn1b) that encodes a protein with a predicted primary structure that is similar to FNs identified in other vertebrates possessing 12 type I, 2 type II and 17 type III repeats including two alternative splice sites (EIIIA and EIIIB) and a variable region (V). Zebrafish FN1b exhibits 62.0% amino acid identity with zebrafish FN1a, 54% with human and 55% with Xenopus laevis FNs respectively. Employing RT-PCR analysis, we demonstrate that EIIIB- and V- isoforms are produced by alternative splicing of a single fn1b transcript. The FN1b EIIIA- isoform was not detected in zebrafish embryos or adult tissues nor were EIIIA, EIIIB or V region splice variants of fn1a found. FN1b mRNA was detected by RT-PCR in embryos at the gastrula-stage (8hpf) through 72hpf and in various adult tissues. EIIIB- and V+ are the predominant forms of FN1b present in the zebrafish embryo. Unlike FN1a, which is present at a relatively high amount at the embryonic stages before gastrulation, the FN1b isoforms are present at very low amounts at the early cleavage stage. The presence of multiple isoforms of FN1b along with a different pattern of expression compared to FN1a indicates that the two fn genes have separate roles in zebrafish development. PMID- 15749004 TI - Impact of setup uncertainty in the dosimetry of prostate and surrounding tissues in prostate cancer patients treated with Peacock/IMRT. AB - The purpose of this paper was to assess the effect of setup uncertainty on dosimetry of prostate, seminal vesicles, bladder, rectum, and colon in prostate cancer patients treated with Peacock intensity-modulated radiation therapy (IMRT). Ten patients underwent computed tomography (CT) scans using the "prostate box" for external, and an "endorectal balloon" for target immobilization devices, and treatment plans were generated (T1). A maximum of +/-5-mm setup error was chosen to model dosimetric effects. Isodose lines from the T1 treatment plan were then superimposed on each patient's CT anatomy shifted by 5 mm toward the cephalad and caudal direction, generating 2 more dosimetric plans (H1 and H2, respectively). Average mean doses ranged from 74.5 to 74.92 Gy for prostate and 73.65 to 74.94 Gy for seminal vesicles. Average percent target volume below 70 Gy increased significantly for seminal vesicles, from 0.53% to 6.26%, but minimally for prostate, from 2.08% to 4.4%. Dose statistics adhered to prescription limits for normal tissues. Setup uncertainty had minimum impact on target dose escalation and normal tissue dosing. The impact of target dose inhomogeneity is currently evaluated in clinical studies. PMID- 15749005 TI - Interfractional dose variation due to seromas in radiotherapy of breast cancer. AB - We investigated the interfractional dose variation due to seromas in radiotherapy of breast cancer patients. For 3 patients who received seroma aspiration during the period of radiotherapy, 2 sets of computed tomography (CT) scanning images were obtained before and after seroma aspiration. Three sets of plans employing a conventional technique and an intensity-modulated radiotherapy (IMRT) technique were generated: the first set of plans was the optimal plan for the CT images before seroma aspiration, the second set was the plans that applied the treatment parameters with the first plans but used CT images obtained after seroma treatment, and the third set was the optimal plans for CT images taken after seroma treatment. From the analysis of each set of plans, we found that the patient anatomy change had little effect on the prescription dose (1-2% variation for both techniques) but had significant effects on the dose homogeneity in the treatment volume, which increased the dose inhomogeneity up to 13.9% for conventional treatment and 20.7% for IMRT treatment, respectively. PMID- 15749006 TI - An assessment of biologically-based optimization (BORT) in the IMRT era. AB - The purpose of this study was to investigate the role of biological-based IMRT (BORT) in treatment planning development; more specifically, to assess the possible advantages of BORT over the classic 3-dimensional conformal radiation therapy (3DRT) and dose-based IMRT based on quantitative and qualitative indices. Three clinical cases are presented to evaluate the differences of BORT, IMRT, and 3DRT. 3DRT, IMRT, and BORT plans were generated for each case using a commercially available treatment planning system (Pinnacle by Philips). The plans were compared by evaluating biological endpoints such as tumor control probability (TCP), normal tissue control probability (NTCP), and uncomplicated tumor control probability (P+), as well as isodose line distribution, dose-volume histograms (DVHs), and dose uniformity. In all cases of this study, BORT yielded improved isodose coverage and P+. Our preliminary results suggest that BORT could play an important role in treatment planning optimization, especially as biological models and predictive assays become more accurate. Further case studies are needed to establish a definitive role for this type of optimization. PMID- 15749007 TI - Comparison of manual vs. automated multimodality (CT-MRI) image registration for brain tumors. AB - Computed tomgoraphy-magnetic resonance imaging (CT-MRI) registrations are routinely used for target-volume delineation of brain tumors. We clinically use 2 software packages based on manual operation and 1 automated package with 2 different algorithms: chamfer matching using bony structures, and mutual information using intensity patterns. In all registration algorithms, a minimum of 3 pairs of identical anatomical and preferably noncoplanar landmarks is used on each of the 2 image sets. In manual registration, the program registers these points and links the image sets using a 3-dimensional (3D) transformation. In automated registration, the 3 landmarks are used as an initial starting point and further processing is done to complete the registration. Using our registration packages, registration of CT and MRI was performed on 10 patients. We scored the results of each registration set based on the amount of time spent, the accuracy reported by the software, and a final evaluation. We evaluated each software program by measuring the residual error between "matched" points on the right and left globes and the posterior fossa for fused image slices. In general, manual registration showed higher misalignment between corresponding points compared to automated registration using intensity matching. This error had no directional dependence and was, most of the time, larger for a larger structure in both registration techniques. Automated algorithm based on intensity matching also gave the best results in terms of registration accuracy, irrespective of whether or not the initial landmarks were chosen carefully, when compared to that done using bone matching algorithm. Intensity-matching algorithm required the least amount of user-time and provided better accuracy. PMID- 15749008 TI - Rectal wall sparing by dosimetric effect of rectal balloon used during intensity modulated radiation therapy (IMRT) for prostate cancer. AB - The use of an air-filled rectal balloon has been shown to decrease prostate motion during prostate radiotherapy. However, the perturbation of radiation dose near the air-tissue interfaces has raised clinical concerns of underdosing the prostate gland. The aim of this study was to investigate the dosimetric effects of an air-filled rectal balloon on the rectal wall/mucosa and prostate gland. Clinical rectal toxicity and dose-volume histogram (DVH) were also assessed to evaluate for any correlation. A film phantom was constructed to simulate the 4-cm diameter air cavity created by a rectal balloon. Kodak XV2 films were utilized to measure and compare dose distribution with and without air cavity. To study the effect in a typical clinical situation, the phantom was computed tomography (CT) scanned on a Siemens DR CT scanner for intensity-modulated radiation therapy (IMRT) treatment planning. A target object was drawn on the phantom CT images to simulate the treatment of prostate cancer. Because patients were treated in prone position, the air cavity was situated superiorly to the target. The treatment used a serial tomotherapy technique with the Multivane Intensity Modulating Collimator (MIMiC) in arc treatment mode. Rectal toxicity was assessed in 116 patients treated with IMRT to a mean dose of 76 Gy over 35 fractions (2.17-Gy fraction size). They were treated in the prone position, immobilized using a Vac Loktrade mark bag and carrier-box system. Rectal balloon inflated with 100 cc of air was used for prostate gland immobilization during daily treatment. Rectal toxicity was assessed using modifications of the Radiation Therapy Oncology Group (RTOG) and late effects Normal Tissue Task Force (LENT) scales systems. DVH of the rectum was also evaluated. From film dosimetry, there was a dose reduction at the distal air-tissue interface as much as 60% compared with the same geometry without the air cavity for 15-MV photon beam and 2x2-cm field size. The dose beyond the interface recovered quickly and the dose reductions due to air cavity were 50%, 28%, 11%, and 1% at 2, 5, 10, and 15 mm, respectively, from the distal air-tissue interface. Evaluating the dose profiles of the more clinically relevant situation revealed the dose at air-tissue interface was approximately 15% lower in comparison to that without an air cavity. The dose built up rapidly so that at 1 and 2 mm, there was only an 8% and 5% differential, respectively. The dosimetric coverage at the depth of the posterior prostate wall was essentially equal with or without the air cavity. The median follow-up was 31.3 months. Rectal toxicity profile was very favorable: 81% (94/116) patients had no rectal complaint while 10.3% (12/116), 6.9% (8/116), and 1.7% (2/116) had grade 1, 2, and 3 toxicity, respectively. There was no grade 4 rectal toxicity. DVH analysis revealed that none of the patients had more than 25% of the rectum receiving 70 Gy or greater. Rectal balloon has rendered anterior rectal wall sparing by its dosimetric effects. In addition, it has reduced rectal volume, especially posterior and lateral rectal wall receiving high-dose radiation by rectal wall distension. Both factors may have contributed to decreased rectal toxicity achieved by IMRT despite dose escalation and higher than conventional fraction size. The findings have clinical significance for future very high-dose escalation trials whereby radiation proctitis is a major limiting factor. PMID- 15749009 TI - Support of large breasts during tangential irradiation using a micro-shell and minimizing the skin dose--a pilot study. AB - Tangential radiotherapy delivered to women with large breasts can be problematic due to the excessive skin folds and the way that the breast falls into the axilla. This may necessitate excessive lung irradiation to cover the posterior part of the breast volume adequately. Conventional breast rings used to move the breast anteriorly can be very difficult to reproduce and may substantially increase the skin dose and hence skin toxicity due to the bolus effect. An in house designed microshell device was constructed to improve setup reproducibility and minimize skin dose. Dose comparisons using a phantom were made between this device and 2 other commonly used devices. The microshell successfully reduced the surface dose compared to the other breast rings tested. This device was then investigated on 8 patients under clinical conditions. Skin doses measured on the trial patients were within acceptable limits. During this small pilot study, no patients suffered excessive skin toxicity or required treatment breaks. Due to the microshell's expandable nature, ease of application, which increases patient comfort compared to other breast rings, and the lower surface dose, the microshell is the preferred breast stabilization device for this department when treating patients with large pendulous breasts. We encourage other departments to consider their current method of breast stabilization and compare them to our results. PMID- 15749010 TI - Setup errors in patients treated with intensity-modulated whole pelvic radiation therapy for gynecological malignancies. AB - Intensity-modulated whole pelvic radiation therapy (IM-WPRT) has decreased the incidence of gastrointestinal complications by reducing the volume of normal tissue irradiated in gynecologic patients. However, IM-WPRT plans result in steep dose gradients around the target volume, and thus accurate patient setup is essential. To quantify the accuracy of our patient positioning, we examined the weekly portal films of 46 women treated with IM-WPRT at our institution. All patients were positioned using a customized immobilization device that was indexed to the treatment table. Setup errors were evaluated by comparing portal images to simulation images using an algorithm that registers user-defined open curve segments drawn on both sets of film. The setup errors, which were separated into systematic and random components, ranged from 1.9 to 3.7 mm for the translations and 1.3 degrees to 4.4 degrees for the 2 in-plane translations. The systematic errors were all less than the respective random errors, with the largest error in the anterior/posterior direction. In addition, there was no correlation between the magnitude of these errors and patient-specific factors (age, weight, height). In the future, we will investigate the effect of these setup errors on the delivered dose distribution. PMID- 15749011 TI - SRprises along a messenger's journey. AB - SR (serine/arginine-rich) proteins were originally identified as essential splicing factors. Recent work, however, has implicated these proteins in numerous additional steps of mRNA metabolism, including nuclear export, RNA stability, mRNA quality control, and translation. PMID- 15749012 TI - The dynamics of histone H1 function in chromatin. AB - Over 80% of the nucleosomes in chromatin contain histone H1, a protein family known to affect the structure and activity of chromatin. Genetic studies and in vivo imaging experiments are changing the traditional view of H1 function and mechanism of action. H1 variants are partially redundant, mobile molecules that interact with nucleosomes as members of a dynamic protein network and serve as fine tuners of chromatin function. PMID- 15749014 TI - Dissecting the mechanism and assembly of a complex virulence mycobacterial lipid. AB - Mycobacterium tuberculosis cell envelope is a treasure house of biologically active lipids of fascinating molecular architecture. Although genetic studies have alluded to an array of genes in biosynthesis of complex lipids, their mechanistic, structural, and biochemical principles have not been investigated. Here, we have dissected the molecular logic underlying the biosynthesis of a virulence lipid phthiocerol dimycocerosate (PDIM). Cell-free reconstitution studies demonstrate that polyketide synthases, which are usually involved in the biosynthesis of secondary metabolites, are responsible for generating complex lipids in mycobacteria. We show that PapA5 protein directly transfers the protein bound mycocerosic acid analogs on phthiocerol to catalyze the final esterification step. Based on precise identification of biological functions of proteins from Pps cluster, we have rationally produced a nonmethylated variant of mycocerosate esters. Apart from elucidating mechanisms that generate chemical heterogeneity with PDIMs, this study also presents an attractive approach to explore host-pathogen interactions by altering mycobacterial surface coat. PMID- 15749013 TI - TbMP42, a protein component of the RNA editing complex in African trypanosomes, has endo-exoribonuclease activity. AB - RNA editing in trypanosomatids is catalyzed by a high molecular mass RNP complex, which is only partially characterized. TbMP42 is a 42 kDa protein of unknown function that copurifies with the editing complex. The polypeptide is characterized by two Zn fingers and a potential barrel structure/OB-fold at its C terminus. Using recombinant TbMP42, we show that the protein can bind to dsRNA and dsDNA but fails to recognize DNA/RNA hybrids. rTbMP42 degrades ssRNA by a 3' to 5' exoribonuclease activity. In addition, rTbMP42 has endoribonuclease activity, which preferentially hydrolyzes non-base-paired uridylate-containing sequences. Gene silencing of TbMP42 inhibits cell growth and is ultimately lethal to the parasite. Mitochondrial extracts from TbMP42-minus trypanosomes have only residual RNA editing activity and strongly reduced endo-exoribonuclease activity. However, all three activities can be restored by the addition of rTbMP42. Together, the data suggest that TbMP42 contributes both endo- and exoribonuclease activity to the editing reaction cycle. PMID- 15749015 TI - 20S proteasomal degradation of ornithine decarboxylase is regulated by NQO1. AB - Ornithine decarboxylase (ODC), a key enzyme in the biosynthesis of polyamines, is a very labile protein. ODC is a homodimeric enzyme that undergoes ubiquitin independent proteasomal degradation via direct interaction with antizyme, a polyamine-induced protein. Binding of antizyme promotes the dissociation of ODC homodimers and marks ODC for degradation by the 26S proteasomes. We describe here an alternative pathway for ODC degradation that is regulated by NAD(P)H quinone oxidoreductase 1 (NQO1). We show that NQO1 binds and stabilizes ODC. Dicoumarol, an inhibitor of NQO1, dissociates ODC-NQO1 interaction and enhances ubiquitin independent ODC proteasomal degradation. We further show that dicoumarol sensitizes ODC monomers to proteasomal degradation in an antizyme-independent manner. This process of NQO1-regulated ODC degradation was recapitulated in vitro by using purified 20S proteasomes. Finally, we show that the regulation of ODC stability by NQO1 is especially prominent under oxidative stress. Our findings assign to NQO1 a role in regulating ubiquitin-independent degradation of ODC by the 20S proteasomes. PMID- 15749016 TI - The molecular architecture of the mammalian DNA repair enzyme, polynucleotide kinase. AB - Mammalian polynucleotide kinase (PNK) is a key component of both the base excision repair (BER) and nonhomologous end-joining (NHEJ) DNA repair pathways. PNK acts as a 5'-kinase/3'-phosphatase to create 5'-phosphate/3'-hydroxyl termini, which are a necessary prerequisite for ligation during repair. PNK is recruited to repair complexes through interactions between its N-terminal FHA domain and phosphorylated components of either pathway. Here, we describe the crystal structure of intact mammalian PNK and a structure of the PNK FHA bound to a cognate phosphopeptide. The kinase domain has a broad substrate binding pocket, which preferentially recognizes double-stranded substrates with recessed 5' termini. In contrast, the phosphatase domain efficiently dephosphorylates single stranded 3'-phospho termini as well as double-stranded substrates. The FHA domain is linked to the kinase/phosphatase catalytic domain by a flexible tether, and it exhibits a mode of target selection based on electrostatic complementarity between the binding surface and the phosphothreonine peptide. PMID- 15749017 TI - Structure of the human telomerase RNA pseudoknot reveals conserved tertiary interactions essential for function. AB - Human telomerase contains a 451 nt RNA (hTR) and several proteins, including a specialized reverse transcriptase (hTERT). The 5' half of hTR comprises the pseudoknot (core) domain, which includes the RNA template for telomere synthesis and a highly conserved pseudoknot that is required for telomerase activity. The solution structure of this essential pseudoknot, presented here, reveals an extended triple helix surrounding the helical junction. The network of tertiary interactions explains the phylogenetic sequence conservation and existing human and mouse TR functional studies as well as mutations linked to disease. Thermodynamic stability, dimerization potential, and telomerase activity of mutant RNAs that alter the tertiary contacts were investigated. Telomerase activity is strongly correlated with tertiary structure stability, whereas there is no correlation with dimerization potential of the pseudoknot. These studies reveal that a conserved pseudoknot tertiary structure is required for telomerase activity. PMID- 15749018 TI - Mediator requirement for both recruitment and postrecruitment steps in transcription initiation. AB - Mediator complexes are required for activators to stimulate Pol II preinitiation complex assembly on an associated promoter. We show here that for the mouse Egr1 gene, controlled largely by MAP kinase phosphorylation of the ELK1 transcription factor, the MED23 Mediator subunit that interacts with phospho-ELK1 is also required to stimulate Pol II initiation at a step subsequent to preinitiation complex assembly. In Med23-/- cells, histone acetylation, methylation, and chromatin remodeling complex association at the Egr1 promoter were equivalent to that of wild-type cells, yet Egr1 induction was greatly reduced. MAP kinase activation stimulated Pol II and GTF promoter binding. However, the difference in factor binding between wild-type and mutant cells was much less than the difference in transcription, and Pol II remained localized to the promoter in mutant cells. These results indicate that an interaction with MED23 stimulates initiation by promoter bound Pol II in addition to Pol II and GTF recruitment. PMID- 15749019 TI - GMP synthetase stimulates histone H2B deubiquitylation by the epigenetic silencer USP7. AB - The packaging of eukaryotic genomic DNA into chromatin is modulated through a range of posttranslational histone modifications. Among these, the role of histone ubiquitylation remains poorly understood. Here, we show that the essential Drosophila ubiquitin-specific protease 7 (USP7) contributes to epigenetic silencing of homeotic genes by Polycomb (Pc). We purified USP7 from embryo nuclear extracts as a stable heteromeric complex with guanosine 5' monophosphate synthetase (GMPS). The USP7-GMPS complex catalyzed the selective deubiquitylation of histone H2B, but not H2A. Biochemical assays confirmed the tight association between USP7 and GMPS in Drosophila embryo extracts. Similar to USP7, mutations in GMPS acted as enhancers of Pc in vivo. USP7 binding to GMPS was required for histone H2B deubiquitylation and strongly augmented deubiquitylation of the human tumor suppressor p53. Thus, GMPS can regulate the activity of a ubiquitin protease. Collectively, these results implicate a biosynthetic enzyme in chromatin control via ubiquitin regulation. PMID- 15749020 TI - A biological network in Saccharomyces cerevisiae prevents the deleterious effects of endogenous oxidative DNA damage. AB - In this study, we used Saccharomyces cerevisiae to identify a biological network that prevents the deleterious effects of endogenous reactive oxygen species. The absence of Tsa1, a key peroxiredoxin, caused increased rates of mutations, chromosomal rearrangements, and recombination. Defects in recombinational DNA double strand break repair, Rad6-mediated postreplicative repair, and DNA damage and replication checkpoints caused growth defects or lethality in the absence of Tsa1. In addition, the mutator phenotypes caused by a tsa1 mutation were significantly aggravated by defects in Ogg1, mismatch repair, or checkpoints. These results indicate that increased endogenous oxidative stress has broad effects on genome stability and is highly sensitive to the functional state of DNA repair and checkpoints. These findings may provide insight in understanding the consequences of various pathophysiological processes in regard to genomic instability. PMID- 15749021 TI - A role for PML3 in centrosome duplication and genome stability. AB - The promyelocytic leukemia gene (PML), which is disrupted by the chromosomal translocation t(15;17) in acute promyelocytic leukemia (APL), encodes a multifunctional protein involved in several important cellular functions. Herein, we demonstrate that PML is localized to centrosomes and that PML deficiency leads to centrosome amplification. By using PML isoform-specific antibodies, we found PML3-specific association with the centrosome and the pole of the mitotic spindle. PML3 deficiency leads to dysregulation of the centrosome duplication checkpoint. Furthermore, PML3 physically interacts with Aurora A and regulates its kinase activity. Specific knockdown of PML3 activates Cdk2/cyclin kinase activity. The results of this study implicate a direct role for PML3 in the control of centrosome duplication through suppression of Aurora A activation to prevent centrosome reduplication. PMID- 15749022 TI - The disposition of nascent strands at stalled replication forks dictates the pathway of replisome loading during restart. AB - Rescue of arrested and collapsed replication forks is essential for maintenance of genomic integrity. One system for origin of replication-independent loading of the DnaB replicative helicase and subsequent replisome reassembly requires the structure-specific recognition factor PriA and the assembly factors PriB and DnaT. Here, we provide biochemical evidence for an alternate system for DnaB loading that requires only PriC. Furthermore, the choice of which system is utilized during restart is dictated by the nature of the structure of the stalled replication fork. PriA-dependent reactions are most robust on fork structures with no gaps in the leading strand, such as is found at the junction of a D loop, while the PriC-dependent system preferentially utilizes fork structures with large gaps in the leading strand. These observations suggest that the type of initial damage on the DNA template and how the inactivated fork is processed ultimately influence the choice of enzymatic restart pathway. PMID- 15749023 TI - Direct visualization of RecBCD movement reveals cotranslocation of the RecD motor after chi recognition. AB - In Escherichia coli, chi (5'-GCTGGTGG-3') is a recombination hotspot recognized by the RecBCD enzyme. Recognition of chi reduces both nuclease activity and translocation speed of RecBCD and activates RecA-loading ability. RecBCD has two motor subunits, RecB and RecD, which act simultaneously but independently. A longstanding hypothesis to explain the changes elicited by chi interaction has been "ejection" of the RecD motor from the holoenzyme at chi. To test this proposal, we visualized individual RecBCD molecules labeled via RecD with a fluorescent nanoparticle. We could directly see these labeled, single molecules of RecBCD moving at up to 1835 bp/s (approximately 0.6 microm/s). Those enzymes translocated to chi, paused, and continued at reduced velocity, without loss of RecD. We conclude that chi interaction induces a conformational change, resulting from binding of chi to RecC, and not from RecD ejection. This change is responsible for alteration of RecBCD function that persists for the duration of DNA translocation. PMID- 15749024 TI - The interleukin-1-related cytokine IL-1F8 is expressed in glial cells, but fails to induce IL-1beta signalling responses. AB - The putative new interleukin (IL)-1 family member IL-1F8 (IL-1eta, IL-1H2) has been shown recently to activate mitogen activated protein kinases (MAPKs), extracellular signal-regulated protein kinase (ERK1/2) and c-Jun N-terminal kinase (JNK), and nuclear factor-kappa B (NFkappa B) via a mechanism that requires IL-1Rrp2 expression in cell lines. The aim of this study was to test the hypothesis that IL-1F8 contributes to brain inflammation and injury, by studying its expression and actions in the different cell types of the mouse brain in culture. Messenger RNA for IL-1F8 was detected in neurons and glia (microglial cells, oligodendrocytes progenitor cells and to a lesser extent astrocytes) by RT PCR. Bacterial lipopolysaccharide (LPS) had no effect on IL-1F8 mRNA levels in mixed glial cultures. Recombinant mouse IL-1beta induced strong activation of ERK1/2, p38, JNK and NFkappa B, and significant release of IL-6 and PGE2, which was blocked by IL-1ra. In contrast, recombinant mouse IL-1F8 did not influence any of these parameters. These results demonstrate that CNS cells may be a source of IL-1F8, but the failure of LPS to modulate IL-1F8 mRNA expression, and of recombinant IL-1F8 to induce any of the classical IL-1 responses, suggest that this cytokine has restricted activities in the brain, or that it may act via alternative pathway(s). PMID- 15749025 TI - Adiponectin expression in human epicardial adipose tissue in vivo is lower in patients with coronary artery disease. AB - BACKGROUND: Intra-peritoneal adipose tissue is recognized as a predictor of metabolic syndrome and may contribute to the risk for cardiovascular disease by the production of adipocytokines, including adiponectin. Nevertheless, there is no knowledge on whether other visceral depots of adipose tissue, including the epicardial fat, have any metabolically active role, including production of adiponectin. AIM OF THE STUDY: We sought to evaluate adiponectin protein expression in epicardial adipose tissue in vivo both in patients with severe coronary artery disease (CAD) and in subjects without CAD. METHODS: Twenty-two patients were enrolled for the study. We selected 16 patients who underwent elective coronary artery bypass graft surgery for critical CAD, 5 who underwent surgery for valve replacement and 1 for correction of an interatrial defect. Epicardial adipose tissue biopsy samples were obtained before the initiation of cardiopulmonary bypass. Adiponectin protein level in epicardial adipose tissue was evaluated by Western blotting. RESULTS: Adiponectin protein value, expressed as adiponectin/actin ratio, in epicardial adipose tissue was significantly lower in patients with severe CAD than in those without CAD (1.42 +/- 0.77 vs 2.36 +/- 0.84 p = 0.02, 95% CI 0.64-1.74). CONCLUSIONS: This study showed for the first time that human epicardial adipose tissue expresses adiponectin. Adiponectin expression is significantly lower in epicardial fat isolated from patients with CAD. PMID- 15749026 TI - Microarray analysis of primary endothelial cells challenged with different inflammatory and immune cytokines. AB - To investigate the potential molecular mediators of tissue-specific recruitment, we explored the influence of different cytokine challenges on gene expression regulation in five primary endothelial cells (ECs), representing two different phenotypes: iliac artery and aortic (macrovascular); lung, colon and dermal (microvascular). We challenged ECs with cytokines that elicit different patterns of inflammatory and immune responses in immune cells: tumor necrosis factor (TNF alpha), interferon-gamma (IFN-gamma) or interleukin-4 (IL-4), and used microarrays containing approximately 40,000 unique cDNAs, to assess changes in differential gene expression relative to untreated cells. Five hundred and sixty three sequences changed by at least 2.5 fold in one or more of the 15 possible EC /cytokine combinations. The list included highly regulated adhesion molecules, chemokines, cytokines, metalloproteases, and IFN-gamma-induced genes. Overall, IFN-gamma caused the largest number of gene expression changes and its profile was least correlated with IL-4. In addition to clusters that were predominantly EC/cytokine specific, we also observed several clusters that were regulated by more than one cytokine across several ECs. Furthermore, we identified genes that were reciprocally expressed in response to different cytokines that could serve as markers of inflammatory and immune expression. These results confirm the importance of microenvironment in primary ECs that could have important applications in developing targeted therapies for vascular diseases. PMID- 15749027 TI - Adipose tissue is a regulated source of interleukin-10. AB - White adipose tissue (WAT) is the source of pro- and anti-inflammatory cytokines and we have recently shown that this tissue is a major source of the anti inflammatory interleukin (IL)-1 receptor antagonist (IL-1Ra). We now aimed at identifying additional adipose-derived cytokines, which might serve as regulators of IL-1Ra. We demonstrate here for the first time that the antiinflammatory cytokine IL-10 is secreted by human WAT explants and that it is up-regulated by LPS and TNF-alpha in vitro, as well as in obesity in humans (2- and 6-fold increase in subcutaneous and visceral WAT, respectively) and rodents (4-fold increase). PMID- 15749029 TI - Negligible induction of IFN-gamma, IL-12 and TNF-alpha by DNA-PEI 750 kDa/albumin complexes. AB - A 750 kDa polyethylenimine (PEI 750 kDa) combined with albumin has been found to mediate in vivo a highly efficient transfection of small amounts of plasmid DNA. Using this exceptional carrier system we evaluated the inflammatory responses triggered by CpG sequences found in plasmid DNA. Using as little as 1 mug DNA transferred in vivo caused an almost negligible response from pro-inflammatory cytokines (IFN-gamma, IL-12 and TNF-alpha), as assessed in serum with a commercially available kit. Administering 750 kDa PEI/albumin/plasmid DNA complexes every three days assured a high and prolonged in vivo expression of a reporter protein. A further increase in the level of such protein was obtained by administering the investigated complexes concurrently with dexamethasone. High gene transfer capability and a relatively low pro-inflammatory response of 750 kDa PEI/albumin/DNA complexes can be exploited for recurrent gene transfer into lungs to treat (via inhalation or instillation) cancer or genetic disorders such as cystic fibrosis. PMID- 15749028 TI - IL-8 promotes cell proliferation and migration through metalloproteinase-cleavage proHB-EGF in human colon carcinoma cells. AB - Interleukin-8 (IL-8) has been reported to promote tumor cell growth in colon cancer cells after binding to its receptors, which are members of the G-protein coupled receptor (GPCR) family. Recent studies demonstrated that stimulation of GPCR can induce shedding of epidermal growth factor (EGF) ligands via activation of a disintegrin and metalloprotease (ADAM), with subsequent transactivation of the EGF receptor (EGFR). In this study, we investigated mechanisms of cell proliferation and migration stimulated by IL-8 in a human colon carcinoma cell line (Caco2). IL-8 increased DNA synthesis of Caco2 in a dose dependent manner and this was inhibited by ADAM, EGFR kinase, and MEK inhibitors. IL-8 transiently induced EGFR tyrosine phosphorylation after 5-90 min and this was completely inhibited by ADAM inhibitor. Neutralizing antibody against HB-EGF as a key ligand for EGFR also blocked transactivation of EGFR and cell proliferation by IL-8. Since IL-8-induced cell migration was further suppressed by the ADAM inhibitor and the HB-EGF neutralizing antibody, our data indicate that IL-8 induces cell proliferation and migration by an ADAM-dependent pathway, and that HB-EGF plays an important role as the major ligand for this pathway. PMID- 15749030 TI - There is more to preventing stroke after carotid surgery than shunt and patch debates. PMID- 15749031 TI - Recurrent carotid artery stenosis following endarterectomy: natural history and risk factors. AB - OBJECTIVES: To establish the incidence of restenosis (RES) following carotid endarterectomy (CEA) and evaluate clinical and technical factors related to its development. DESIGN: Prospective non-randomised cohort study. PATIENTS AND METHODS: Two hundred and twenty-four patients with 243 CEA between May 1998 and December 2002, were subjected to clinical and haemodynamic follow-up, median follow-up 23 months (1-56). There was selective use of a shunt (17.3%) and patch (61.7%). RES (> or =50%) and severe restenosis, > or =70%, (sRES) were defined as peak systolic velocities of > or =150 and > or =300cm/s (or > or =250cm/s with diastolic velocity >100cm/s), respectively. Rates of RES, symptom development and mortality were analysed using Kaplan-Meier curves. Cox's regression model (hazards ratio/95% CI) was used to evaluate prognostic factors. RESULTS: We detected 13 sRES (5.3%) (median time 6.1 months) and 30 (12.3%) moderate stenosis (mRES) (median time 3.7 months). Cumulative freedom from sRES at 23 months was 94.2%. Five sRES detected in the first 45 days after the procedure were deemed to be residual restenosis (rRES). Five (38.4%) sRES were symptomatic, 15.3% progressed to occlusion. Patient survival was 98.0 and 96.4% at 12 and 24 months, respectively. Independent risk factors for sRES: female sex (HR: 3.3, 95% CI 1.1 10 p=0.04) and diabetes (HR: 4.5, 95% CI 1.4-13.9 p=0.008). CONCLUSIONS: Carotid restenosis appears early, is usually low-grade and mostly asymptomatic. Although few stenoses progress to occlusion, women and diabetic patients were at highest risk. PMID- 15749032 TI - Long term outcome of transcranial bypass. AB - OBJECTIVES: This study examines the long term outcome of transcranial bypass in a single centre. METHODS: In a prospective study, symptoms and angiographic findings were recoded on a database for 229 consecutive transcranial bypass procedures performed in 204 patients between 1976 and 1983. One hundred and seventy-seven had carotid territory symptoms, and the predominant angiogram finding was internal carotid artery (ICA) occlusion. RESULTS: At 5 years 87% (86 patients) were free from stroke. At 20 years 83% (29 patients) were stroke free but by this time the majority had died from other cardiovascular causes and 35 were lost to follow up. CONCLUSION: Stroke is uncommon five or more years after transcranial bypass but overall survival is poor. PMID- 15749033 TI - Combined open and endovascular stent grafting of internal carotid artery fibromuscular dysplasia: long term results. AB - BACKGROUND: Symptomatic fibromuscular dysplasia (FMD) of the internal carotid artery (ICA) can present as thrombo-embolic ischemic events, spontaneous or post traumatic dissection, aneurysmal degeneration or intracranial haemorrhage and needs definitive surgical treatment. PATIENTS AND METHODS: Six patients and nine ICA with FMD were revascularised using a carotid approach with minimal exposure of the common, external and internal carotid arteries for covered stent repair. All patients were female, the age ranged from 30 to 65 years (mean 44). RESULTS: One patient suffered from a perioperative transient neurological deficit. Duplex revealed a patent stent. The patient fully recovered after 5h, not showing any changes on repeat CT scans. One patient developed a recurrent laryngeal nerve palsy. The symptoms gradually resolved within 1 month. No perioperative strokes or deaths occurred. During a mean follow up of 48 months (range 13-63) no thromboembolic neurological events, graft occlusions or haemodynamically significant stenoses occurred. CONCLUSION: ICA FMD stent grafting is an alternative to open surgery or percutaneous endovascular intervention with excellent long-term results. PMID- 15749034 TI - Primary stenting for the acute treatment of carotid artery dissection. AB - OBJECTIVES: To review and evaluate our experience with carotid artery stenting in the acute treatment of carotid artery dissection (CAD). PATIENTS AND METHODS: Reviewing the charts of our hospital between 2000 and 2001, we found two consecutive patients who benefited from primary stenting for the acute treatment of spontaneous extracranial internal CAD. RESULTS: Primary stenting of the internal carotid artery was successful in both cases without any post-operative complications. Clinical and US duplex scan follow-up confirmed the absence of neurological symptoms and the patency of the internal carotid artery with complete disappearance of the dissection at 36 and 42 months after the procedure, respectively. CONCLUSION: Despite the small number of patients, primary stenting for acute CAD seems to be safe and effective. PMID- 15749035 TI - Inflammatory abdominal aortic aneurysms. AB - BACKGROUND: Inflammatory abdominal aortic aneurysms (IAAA) are a variant of aortic aneurysm characterised by extensive peri-aneurysmal fibrosis, thickened walls and dense adhesions and represent between 3 and 10% of all abdominal aortic aneurysms (AAA). Surgery is technically challenging and is still associated with an increased morbidity and mortality. Controversy exists about aetiology and pathogenesis. METHODS: We review the literature on the current theories, the available imaging modalities and the current thinking on management of IAAA. A Medline database search was performed. Articles were cross-referenced. RESULTS AND CONCLUSIONS: Aneurysm development is multifactorial with important genetic and environmental factors. The literature supports the theory that IAAA arise from the same antigenic stimulus that is responsible for the non-IAAA, representing one extreme of an inflammatory spectrum. The results after open repair have improved and there is now little difference in the mortality between non-IAAA and IAAA repair. However, there is likely to be a role for endovascular stenting in IAAA management and this requires further study. It is clear that closer follow-up of patients after IAAA repair with either technique is necessary to monitor the inflammatory process. No evidence-based follow-up protocol exists but three to six-monthly monitoring of renal function and erythrocyte sedimentation rate (ESR) for 24 months post-repair would seem a reasonable regime. PMID- 15749036 TI - Results of endovascular repair of inflammatory abdominal aortic aneurysms. A report from the EUROSTAR database. AB - OBJECTIVES: To investigate the results following endovascular treatment of patients with inflammatory abdominal aortic aneurysms (IAAA). DESIGN: Retrospective study based on the EUROSTAR registry. MATERIAL AND METHODS: Patients included in the EUROSTAR registry with IAAA (n=52, 1.4%) were compared to those having aneurysms without aortic fibrosis (n=3613, 98.6%). The mean follow-up period in patients with IAAA was 23 months (range 1-60). In 11 of the patients detailed information on the effect of endovascular repair and perianeurysmal fibrosis and ureteral entrapment was obtained by a dedicated questionnaire. RESULTS: Twelve patients (23%) with IAAA had preoperative impairment of renal function and five had known hydronephrosis. Variables that were significantly associated with IAAA included younger age (p<.0001, mean difference 5.9, CI 3.7-7.9) and lower pulmonary risks score (OR 0.38, CI 0.19 0.74). At completion of the endovascular procedure, device stenosis was more frequently observed in patients with IAAA (OR 18.1, CI 3.52-93.0). There were no differences with regard to the rates of mortality, rupture or conversion in patients with IAAA and controls. In the majority, the aneurysm size regressed irrespective of nature of aneurysm. Of the 11 patients with a detailed assessment three had deterioration of renal function and three still had ureteral entrapment during follow-up. CONCLUSION: Despite persistence of perianeurysmal inflammation in a proportion of patients operative and midterm results of endovascular repair were comparable in the patients with inflammatory and standard AAA. PMID- 15749037 TI - Systemic dilation diathesis in patients with abdominal aortic aneurysms: a role for matrix metalloproteinase-9? AB - INTRODUCTION: Accumulating evidence suggests that patients with abdominal aortic aneurysm (AAA) suffer from a systemic dilating condition affecting all arteries. Matrix metalloproteinases (MMPs) and their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), appear to be involved in aneurysm formation, as evidenced by increased aortic tissue MMP activity and plasma MMP levels in patients with AAA. Hypothesizing that an imbalance in plasma MMP/TIMP level might be associated with a systemic dilation diathesis, we studied mechanical vessel wall properties of non-affected arteries of patients with either AAA or aorto-iliac obstructive lesions in association with plasma MMP-9 and TIMP-1 levels. METHODS: Twenty-two patients with AAA and 12 with aorto-iliac occlusive disease (AOD) were included. Diastolic diameter (d) and distension (Deltad) were measured at the level of the common carotid artery (CCA) and suprarenal aorta (SA) using ultrasonography. Distensibility (DC) and compliance (CC) were calculated from d, Deltad and brachial pulse pressure. Plasma MMP-9 and TIMP-1 were determined with specific immunoassays. RESULTS: The average (+/-SD) age was 72.3+/-5.6 and 65.0+/-8.2 years for the AAA and AOD patients, respectively, (P=0.005). CCA diameter was 9.1+/-1.3mm in AAA patients and AOD 7.8+/-1.4mm in AOD patients, P=0.009. This difference persisted after correction for age. Plasma MMP-9 and TIMP-1 did not differ significantly between AAA and AOD patients. In the total 34 patients, the MMP-9/TIMP-1 ratio was correlated inversely with distensibility (r=-0.74, P=0.002) and to compliance (r=-0.58, P=0.024) of the suprarenal aorta. CONCLUSIONS: The CCA diameter was larger in AAA patients compared to AOD patients. MMP-9/TIMP-1 ratio was associated with decreased distensibility and compliance of the suprarenal aorta. These data support the idea that AAA patients exhibit a systemic dilation diathesis, which might be attributable to MMP/TIMP imbalances. PMID- 15749039 TI - Fate of the visceral aortic patch after thoracoabdominal aortic repair. AB - OBJECTIVE: To analyse the fate of a visceral aortic patch (VAP) in patients that underwent thoracoabdominal aortic aneurysm (TAAA) repair. METHODS: We reviewed 204 consecutive patients (158 M, 46 F) treated for TAAA between 1988 and 2004. We performed VAP in 182 cases. Among the 149 survivors at 6 months, we followed 138 cases, mean follow-up 7 years (range 0.6-16 years). The mean graft diameter we used was 29mm (range 24-34mm) from 1988 to 1999 (83 patients), and 21.7mm (range 16-24mm) from 2000 to 2003 (55 patients). In 23% of cases we performed a separate bypass to the left renal artery. RESULTS: We observed 16 (12%) VAP dilatations (<5cm), 6 (4%) VAP aneurysms (>5cm) and one VAP pseudoaneurysm, at a mean time of 6 years after atherosclerotic TAAA was atherosclerotic repair. There were no VAP dilatations/aneurysms in the group of patients with separate left renal revascularization. Five VAP aneurysms were treated electively. In four cases the operation was performed with thoracophrenolaparotomy, in one case with a bilateral subcostal laparotomy. In all cases the visceral aorta was re-grafted. Reimplantation of a single undersized VAP was performed in one case, separate revascularization of visceral arteries was performed in the other four cases. Selective intraoperative hypothermic perfusion of visceral and renal arteries was used in all the patients. There was 1 perioperative death; 2 patients with preoperative renal failure required dialysis. The last VAP aneurysm has remained asymptomatic and stable at annual CT surveillance. The VAP pseudoaneurysm was successfully treated with an emergency thoracophrenolaparotomy and refashioning the left side suture line. CONCLUSIONS: Aneurysm of VAP is not uncommon in the patients operated on using larger grafts with a single VAP that includes the LRA (7.4%, 5/67 cases). Its treatment carries significant morbidity and mortality. PMID- 15749038 TI - Aminoterminal propeptide of type III procollagen and matrix metalloproteinases-2 and -9 failed to serve as serum markers for abdominal aortic aneurysm. AB - OBJECTIVES: Matrix-metalloproteinase (MMP)-2 and -9 and aminoterminal propeptide of type III collagen (NIIINP) have been reported to be elevated in patients with abdominal aortic aneurysm (AAA). The aim of our study was to test NIIINP, MMP-2 and -9 as potential serum markers for AAA in a large population group at risk for AAA. METHODS: Fifty-five to 70 year old men were screened for AAA by abdominal ultrasound. Simultaneously, blood samples were taken and the patients were interviewed for known risk factors for AAA. Patients with a dilatation of the infrarenal aorta of > or =25mm (Group 1, n=76) were compared to randomly assigned patients with normal aortic diameters (Group 2, n=83). A third group consisted of patients scheduled for operation of AAA (n=19). RESULTS: A total of 987 men were investigated with ultrasound. Seventy-six (7.7%) had an aortic dilatation > or =25mm. Aortic dilatation was correlated with age (P=0.0001). However, serum levels of NIIINP and MMP 2 were not different between the three groups of patients. For MMP-9 there was a weak inverse correlation with lower serum levels in patients with aortic dilatation (P=0.043). CONCLUSIONS: Both MMP-2 and -9 and NIIINP failed to show relevance as serum markers for aortic dilatation. Our results are, therefore, in contradiction to previous published results. AAAs cannot be diagnosed with a simple blood test. PMID- 15749040 TI - Neutrophil activation occurs in the lower-limbs of patients undergoing elective repair of abdominal aortic aneurysm. AB - OBJECTIVES: AAA repair is associated with a systemic inflammatory response, mediated in part by neutrophils. The aim of this study was to determine where neutrophil activation occurs. METHODS: Blood was sampled from the femoral vein, portal vein and radial artery of 10 patients undergoing elective AAA repair at four time-points [induction of anaesthesia (systemic sample only), pre-aortic clamp application, pre-clamp removal and after 30min of reperfusion]. Whole blood was analysed for the white cell count, neutrophil count, and for neutrophil CD11b expression. RESULTS: The white cell count and neutrophil counts increased after aortic clamp release. Neutrophil expression of CD11b was significantly higher in the femoral vein than the portal vein and systemic circulation during ischaemia [P=0.001 (FV vs. PV), P=0.017 (FV vs. systemic)] and reperfusion [P=0.001 (FV vs. PV), P=0.013 (FV vs. systemic)]. There were no significant differences in neutrophil CD11b expression between the systemic and portal vein samples at any time. CONCLUSIONS: Ischaemia and reperfusion during abdominal aortic aneurysm repair are associated with a global increase in the white cell count and neutrophil count, but with increased neutrophil CD11b expression only in the femoral vein. This suggests the lower-limbs are sensitive to aortic clamp-related reperfusion injury and may fuel the inflammatory response. PMID- 15749041 TI - Heat shock proteins in vascular disease--a review. AB - INTRODUCTION: There is growing evidence that heat shock proteins (HSPs), a family of stress-inducible proteins may be involved in the pathogenesis of atherosclerotic vascular diseases. Here, we systematically review the evidence behind this notion. METHODS: A detailed literature search and extensive bibliographic review of literature relating to HSPs and atherosclerotic vascular disease. RESULTS: Atherosclerotic vascular disease is classified into four main areas of presentation: carotid, coronary, aortic and peripheral vascular disease, for consideration in this review. In each of these vascular diseases, the evidence linking HSPs and atherosclerosis is outlined in a systematic manner. Current evidence suggests that components of the immune system may be involved in the pathogenesis of atherosclerosis, with HSPs acting as auto-antigens in the immune response. HSPs are detected in atherosclerotic lesions and antibodies to HSPs are increased in patients with vascular disease; the rise often correlating with the severity of atherosclerosis. The levels of anti-HSP antibodies have been shown to be independent predictors of risk and have prognostic value. CONCLUSION: There is a strong link between heat shock protein expression and the principal manifestations of atherosclerotic vascular diseases. A better understanding of this involvement could lead to the development of new and improved treatment strategies. PMID- 15749042 TI - Secondary prevention of atherosclerosis through chlamydia pneumoniae eradication (SPACE Trial): a randomised clinical trial in patients with peripheral arterial disease. AB - BACKGROUND: Sero-epidemiological and experimental studies suggest that Chlamydia pneumoniae infections play an important role in the development of atherosclerosis. Clinical trials have shown contradictory results regarding the efficacy of antibiotics to prevent atherosclerosis-related complications in patients with coronary artery disease. Our aim was to study the effect of a short course of azithromycin on the incidence of cardiovascular events and peripheral vascular function in patients with stable peripheral arterial disease (PAD). PATIENTS AND METHODS: Five hundred and nine PAD-patients were randomised to receive either a 3-day course of azithromycin (500 mg daily) or placebo, with 2 years of follow-up. C. pneumoniae serology was determined at baseline. Clinical endpoints were death, coronary events (myocardial infarction, unstable angina, and/or coronary revascularization procedures), cerebral events (stroke, TIA, and/or carotid endarterectomy) and peripheral arterial complications (increased PAD-symptoms with decreased ankle-brachial index (ABPI, 0.1-point decrease after 12 months), and/or peripheral revascularization procedures). RESULTS: Five hundred and nine patients (160 women) with an atherosclerotic risk factor profile were randomised, 257 patients to azithromycin and 252 to placebo. Four hundred and forty nine patients (88%) had intermittent claudication and 60 (12%) had critical limb ischemia. By 24-month follow up, 182 patients (36%) developed 252 complications (45 deaths, 34 coronary events, 34 cerebral events and 139 peripheral arterial complications). C. pneumoniae IgA-titres were associated with the development of cardiovascular events. Nevertheless, the number of complications (131 in the azithromycin group vs. 121 in the placebo group) and the number of patients that developed complications (98 (38%) in the azithromycin vs. 84 (33%) in the placebo group) was comparable in both treatment groups. Life table analysis showed no effect of azithromycin on survival or ABPI. CONCLUSION: A short-term course of azithromycin offers no benefits for survival or ankle pressure in PAD-patients. PMID- 15749043 TI - Antibacterial activity of gentamicin-bonded gelatin-sealed polyethylene terephthalate vascular prostheses. AB - OBJECTIVES: To create an antibiotic-modified vascular prosthesis with a prolonged bactericidal activity, susceptible to endothelialisation. METHODS: We used a covalent method of gentamicin sulphate immobilisation to polyethylene terephthalate prosthesis sealed with gelatin. Antibacterial activity was assayed in Luria-Bertani medium against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa strains. Prosthesis endothelialisation was performed using bovine aorta endothelial cells (BAEC). RESULTS: Gentamicin was bound to vascular prostheses in the amount of 12g per kg of prosthesis. Ninety-seven percent of antibiotic bound in covalent way and remained on the biomaterial for at least 30 days during shaking in PBS solution. Gentamicin-modified prostheses exerted bactericidal or bacteriostatic effect on growth of clinical and reference bacterial strains, prevented biofilm formation and were highly susceptible to endothelialisation. BAEC viability exceeded 90%, which indicated that gentamicin vascular prostheses were not toxic for these cells. CONCLUSIONS: Covalent gentamicin immobilisation resulted in effective antibacterial protection of vascular prostheses against clinical and reference strains of S. aureus, E. coli and P. aeruginosa and allowed for a strong adherence of endothelial cells to antibiotic-modified prostheses. PMID- 15749044 TI - The impact of adjuvant av-fistula on cuffed Femorocrural PTFE bypass grafting: flow and pressure response. AB - OBJECTIVE: The aim of the present study was to assess the effect of an adjuvant av-fistula on prosthetic bypass grafting and whether intraoperative flow measurements could predict patency and adverse events of cuffed femorocrural PTFE bypass with or without an av-fistula. METHODS: A total of 50 patients in need of vascular reconstruction for critical limb ischaemia (CLI) but with no suitable venous conduit were included. RESULTS: The flow values in patients with av fistula were significantly higher (p=0.009) than in the group without the fistula but the higher flow values did not result in improved patency. The maximum flow velocity (Vmax) in the av-fistula group was significantly higher in the immediate postoperative period (p=0.04), but there was no difference in patency. When a flow value of 50 ml/min was used as a cut-off point, patients with a higher flow had significantly better immediate patency (p=0.025). CONCLUSION: The adjuvant av fistula neither caused any adverse effects nor had any effect on patency. PMID- 15749045 TI - Thoracofemoral bypass using spliced femoral vein with removal of an infected axillobifemoral bypass graft. AB - A 63-year-old male underwent emergency repair of a ruptured juxtarenal aortic aneurysm via a transabdominal approach using an aorto-bi-iliac Dacron graft. This became infected. A right axillobifemoral bypass was placed and the infected graft was removed with oversewing of the aorta. The patient was re-admitted 8 months later with an infected axillobifemoral prosthesis. We harvested both femoral veins (FV) and spliced them to perform a left thoracobifemoral bypass with simultaneous explantation of the infected graft. The patient remains well with a patent graft 20 months post-operatively. PMID- 15749046 TI - Pentoxifylline for leg ulcers. PMID- 15749047 TI - RNA silencing as a tool for exploring gene function in ascomycete fungi. AB - We have developed a pHANNIBAL-like silencing vector, pSilent-1, for ascomycete fungi, which carries a hygromycin resistance cassette and a transcriptional unit for hairpin RNA expression with a spacer of a cutinase gene intron from the rice blast fungus Magnaporthe oryzae. In M. oryzae, a silencing vector with the cutinase intron spacer (147 bp) showed a higher efficiency in silencing of the eGFP gene than did those with a spacer of a GUS gene fragment or a longer intron (850 bp) of a chitin binding protein gene. Application of pSilent-1 to two M. oryzae endogenous genes, MPG1 and polyketide synthase-like gene, resulted in various degrees of silencing of the genes in 70-90% of the resulting transformants. RNA silencing was also induced by a pSilent-1-based vector in Colletotrichum lagenarium at a slightly lower efficiency than in M. oryzae, indicating that this silencing vector should provide a useful reverse genetic tool in ascomycete fungi. PMID- 15749048 TI - Nitric oxide has a regulatory effect in the germination of conidia of Colletotrichum coccodes. AB - Nitric oxide (NO) was first detected in mammals and has since been found in plants and in micro-organisms such as bacteria. NO is an important signalling molecule involved in a number of critical signal transduction pathways. To date, NO has not been directly detected in fungi, and little research on NO and fungi has been completed. Here, the role of NO in the germination of Colletotrichum coccodes conidia was investigated. Conidia were germinated on microscope slides, treated with chemicals to block NO, to add NO, and/or to detect NO, and assessed for their stage of development over 24 h. NO was detected in germinating conidia at all stages of development. Exogenous NO delayed germination, while treatment with NO inhibitors accelerated germination, suggesting NO may have a regulatory effect in germination. The differential effect of the various inhibitors suggests the fungal isoform of nitric oxide synthase (NOS) may be biochemically similar to mammalian constitutive NOS. PMID- 15749049 TI - Ligand interactions of the Coprinopsis cinerea galectins. AB - The basidiomycete Coprinopsis cinerea (Coprinus cinereus) expresses two fruiting body-specific isolectins (CGL1 and CGL2) that belong to the family of galectins. Understanding the role of these beta-galactoside binding lectins is still in the beginning. Even though the prerequisites for substrate binding are well understood, it is not known how discrimination between potential substrates is achieved and what kind of influence this has on the function in a distinct cellular context. Precise knowledge of the expression of galectins and their ligands will aid in elucidating their function. In Coprinopsis, the developmentally regulated ligands for galectins co-localise with galectin expression in the veil surrounding the developing primordium and the outer cells of the young stipe. In addition, galectin ligands are observed in the hymenium. The subcellular localisation of the galectin ligands suggests these to be present in cellular compartments distinct from galectin transport. The sensitivity of the in situ interactions with exogenous galectin towards detergents and organic solvents infers that these ligands are lipid-borne. Accordingly, lipid fractions from primordia are shown to contain galectin-binding compounds. Based on these results and the determined binding specificity towards substituted beta galactosides we hypothesise that beta-galactoside-containing lipids (basidiolipids) found in mushrooms are physiological ligands for the galectins in C. cinerea. PMID- 15749050 TI - The tetraspanin gene ClPLS1 is essential for appressorium-mediated penetration of the fungal pathogen Colletotrichum lindemuthianum. AB - Conservation of the molecular mechanisms controlling appressorium-mediated penetration during evolution was assessed through a functional study of the ClPLS1 gene from Colletotrichum lindemuthianum orthologous to the MgPLS1 from Magnaporthe grisea, involved in penetration peg development. These two plant pathogenic Pyrenomycetes differentiate appressoria to penetrate into plant tissues. We showed that ClPLS1 is a functional homologue of MgPLS1 in M. grisea. Loss of ClPLS1 function had no effect on vegetative growth, conidiation or on appressorium differentiation and maturation. However, Clpls1::hph mutants are non pathogenic on either intact or wounded bean leaves, as a result of a defect in the formation and/or positioning of the penetration pore and consequently in the formation of the penetration peg. These observations suggest that the fungal tetraspanins control a conserved appressorial function that could be required for the correct localization of the site where the penetration peg emerges. PMID- 15749051 TI - Identification, cloning, and functional expression of three glutathione transferase genes from Aspergillus fumigatus. AB - Analysis of the genome of the human pathogen, Aspergillus fumigatus, revealed the presence of several putative glutathione transferase (GST) open reading frames. Three A. fumigatus GST genes, termed gstA, B, and C, were cloned and recombinant proteins expressed in Escherichia coli. Functional analysis of recombinant gstA-C confirms that the enzymes exhibit GST activity and glutathione peroxidase activity. RT-PCR confirmed low basal expression of gstA and gstC which was markedly up-regulated (at least 4x-10x) in the presence of either H2O2 or 1 chloro-2,4-dinitrobenzene (CDNB). GstB expression was only observed in the presence of CDNB. These results demonstrate for the first time the existence of three functional GSTs in A. fumigatus and strongly suggest a role for these enzymes in the response of the organism to both oxidative stress and xenobiotic presence. PMID- 15749053 TI - Cloning and analysis of pppg1, an inducible endopolygalacturonase gene from the oomycete plant pathogen Phytophthora parasitica. AB - Phytophthora parasitica is an oomycete plant pathogen that causes severe disease in a wide variety of crops. Here, we report the isolation of a gene, named pppg1, which encodes an extracellular endopolygalacturonase in P. parasitica. Both cDNA and a genomic clone were isolated and sequenced. The pppg1 gene showed standard characteristics with respect to core promoter and intron sequences of Phytophthora. The predicted protein of pppg1 has a calculated molecular mass of 39.7 kDa and a pI value of 5.2, and contains a putative signal peptide of 20 amino acid residues on the N-terminus. The deduced amino acid sequence is highly conserved with those of other Phytophthora and fungal endopolygalacturonases. Analysis by reverse transcription followed by real-time quantitative polymerase chain reaction showed that transcription of pppg1 was repressed by glucose, but induced by pectin in the culture. Moreover, pppg1 is highly expressed during interaction of P. parasitica with the host plant, suggesting its involvement in the process of host infection. Heterologous expression of pppg1 in Pichia pastoris produced proteins with molecular mass ranging from 75 to 200 kDa, very likely due to differential glycosylation by the yeast. Deglycosylation of the recombinant protein resulted in a complete loss of the endopolygalacturonase activity. PMID- 15749052 TI - Functional expression of the Candida albicans alpha-factor receptor in Saccharomyces cerevisiae. AB - Candida albicans genes involved in mating have been identified previously by homology to Saccharomyces cerevisiae mating pathway components. The C. albicans genome encodes CaSte2p, a homolog of the S. cerevisiae alpha-mating pheromone receptor Ste2p, and two potential pheromones, alpha-F13 (GFRLTNFGYFEPG) and alpha F14 (GFRLTNFGYFEPGK). The response of several C. albicans strains to the synthesized peptides was determined. The alpha-F13 was degraded by a C. albicans MTLa strain but not by S. cerevisiae MATa cells. The CaSTE2 gene was cloned and expressed in a ste2-deleted strain of S. cerevisiae. Growth arrest and beta galactosidase activity induced from a FUS1-lacZ reporter construct increased in a dose-dependent manner upon exposure of transgenic S. cerevisiae to alpha-F13. Mating between the strain expressing CaSTE2 and an opposite mating type was mediated by alpha-F13 and not by the S. cerevisiae alpha-factor. The results indicated that CaSte2p effectively coupled to the S. cerevisiae signal transduction pathway. Functional expression of CaSte2p in S. cerevisiae provides a well-defined system for studying the biochemistry and molecular biology of the C. albicans pheromone and its receptor. PMID- 15749054 TI - Families of short interspersed elements in the genome of the oomycete plant pathogen, Phytophthora infestans. AB - The first known families of tRNA-related short interspersed elements (SINEs) in the oomycetes were identified by exploiting the genomic DNA sequence resources for the potato late blight pathogen, Phytophthora infestans. Fifteen families of tRNA-related SINEs, as well as predicted tRNAs, and other possible RNA polymerase III-transcribed sequences were identified. The size of individual elements ranges from 101 to 392 bp, representing sequences present from low (1) to highly abundant (over 2000) copy number in the P. infestans genome, based on quantitative PCR analysis. Putative short direct repeat sequences (6-14 bp) flanking the elements were also identified for eight of the SINEs. Predicted SINEs were named in a series prefixed infSINE (for infestans-SINE). Two SINEs were apparently present as multimers of tRNA-related units; four copies of a related unit for infSINEr, and two unrelated units for infSINEz. Two SINEs, infSINEh and infSINEi, were typically located within 400 bp of each other. These were also the only two elements identified as being actively transcribed in the mycelial stage of P. infestans by RT-PCR. It is possible that infSINEh and infSINEi represent active retrotransposons in P. infestans. Based on the quantitative PCR estimates of copy number for all of the elements identified, tRNA-related SINEs were estimated to comprise 0.3% of the 250 Mb P. infestans genome. InfSINE-related sequences were found to occur in species throughout the genus Phytophthora. However, seven elements were shown to be exclusive to P. infestans. PMID- 15749055 TI - Structure and regulation of acetyl-CoA carboxylase genes of metazoa. AB - Acetyl-CoA carboxylase (ACC) plays a fundamental role in fatty acid metabolism. The reaction product, malonyl-CoA, is both an intermediate in the de novo synthesis of long-chain fatty acids and also a substrate for distinct fatty acyl CoA elongation enzymes. In metazoans, which have evolved energy storage tissues to fuel locomotion and to survive periods of starvation, energy charge sensing at the level of the individual cell plays a role in fuel selection and metabolic orchestration between tissues. In mammals, and probably other metazoans, ACC forms a component of an energy sensor with malonyl-CoA, acting as a signal to reciprocally control the mitochondrial transport step of long-chain fatty acid oxidation through the inhibition of carnitine palmitoyltransferase I (CPT I). To reflect this pivotal role in cell function, ACC is subject to complex regulation. Higher metazoan evolution is associated with the duplication of an ancestral ACC gene, and with organismal complexity, there is an increasing diversity of transcripts from the ACC paraloges with the potential for the existence of several isozymes. This review focuses on the structure of ACC genes and the putative individual roles of their gene products in fatty acid metabolism, taking an evolutionary viewpoint provided by data in genome databases. PMID- 15749056 TI - Function of prokaryotic and eukaryotic ABC proteins in lipid transport. AB - ATP binding cassette (ABC) proteins of both eukaryotic and prokaryotic origins are implicated in the transport of lipids. In humans, members of the ABC protein families A, B, C, D and G are mutated in a number of lipid transport and metabolism disorders, such as Tangier disease, Stargardt syndrome, progressive familial intrahepatic cholestasis, pseudoxanthoma elasticum, adrenoleukodystrophy or sitosterolemia. Studies employing transfection, overexpression, reconstitution, deletion and inhibition indicate the transbilayer transport of endogenous lipids and their analogs by some of these proteins, modulating lipid transbilayer asymmetry. Other proteins appear to be involved in the exposure of specific lipids on the exoplasmic leaflet, allowing their uptake by acceptors and further transport to specific sites. Additionally, lipid transport by ABC proteins is currently being studied in non-human eukaryotes, e.g. in sea urchin, trypanosomatides, arabidopsis and yeast, as well as in prokaryotes such as Escherichia coli and Lactococcus lactis. Here, we review current information about the (putative) role of both pro- and eukaryotic ABC proteins in the various phenomena associated with lipid transport. Besides providing a better understanding of phenomena like lipid metabolism, circulation, multidrug resistance, hormonal processes, fertilization, vision and signalling, studies on pro- and eukaryotic ABC proteins might eventually enable us to put a name on some of the proteins mediating transbilayer lipid transport in various membranes of cells and organelles. It must be emphasized, however, that there are still many uncertainties concerning the functions and mechanisms of ABC proteins interacting with lipids. In particular, further purification and reconstitution experiments with an unambiguous role of ATP hydrolysis are needed to demonstrate a clear involvement of ABC proteins in lipid transbilayer asymmetry. PMID- 15749057 TI - Regulatory enzymes of phosphatidylcholine biosynthesis: a personal perspective. AB - Phosphatidylcholine is a prominent constituent of eukaryotic and some prokaryotic membranes. This Perspective focuses on the two enzymes that regulate its biosynthesis, choline kinase and CTP:phosphocholine cytidylyltransferase. These enzymes are discussed with respect to their molecular properties, isoforms, enzymatic activities, and structures, and the possible molecular mechanisms by which they participate in regulation of phosphatidylcholine levels in the cell. PMID- 15749058 TI - Phosphatidylcholine-rich acceptors, but not native HDL or its apolipoproteins, mobilize cholesterol from cholesterol-rich insoluble components of human atherosclerotic plaques. AB - To examine the potential of high density lipoproteins (HDL) to ameliorate atherosclerotic plaques in vivo, we examined the ability of native HDL, lipid free HDL apolipoproteins (apo HDL), cholesterol-free discoidal reconstituted HDL (R-HDL) comprised of apo HDL and phosphatidylcholine (PC) and PC liposomes to release cholesterol from cholesterol-rich insoluble components of plaques (ICP) isolated from atherosclerotic human aorta. Isolated ICP had a free cholesterol (FC) to phospholipid (PL) mass ratio (0.8-3.1) and a sphingomyelin (SPM) to PC mass ratio (1.2-4.2) that exceeded those of plasma membranes of cultured cells. Surprisingly, native HDL and its apolipoproteins were not able to release cholesterol from ICP. However, R-HDL and PC liposomes were effectively released cholesterol from ICP. The release of ICP cholesterol by R-HDL was dose-dependent and accompanied by the transfer of > 8 x more PC in the reverse direction (i.e., from R-HDL to ICP), resulting in a marked enrichment of ICP with PC. Compared to R-HDL, PC liposomes were significantly less effective in releasing cholesterol from ICP but were somewhat more effective in enriching ICP with PC. Native HDL was minimally effective in enriching ICP with PC, but became effective after prior in vitro enrichment of HDL with PC from multilamellar PC liposomes. The enrichment of ICP with PC resulted in the dissolution of cholesterol crystals on ICP and allowed the removal of ICP cholesterol by apo HDL and plasma. Our study revealed that the removal of cholesterol from ICP in vivo will be possible through a change in the level, composition, and physical state of ICP lipids mediated by PC-enriched HDL. PMID- 15749059 TI - High cholesterol absorption efficiency and rapid biliary secretion of chylomicron remnant cholesterol enhance cholelithogenesis in gallstone-susceptible mice. AB - The study of chylomicron pathway through which it exerts its metabolic effects on biliary cholesterol secretion is crucial for understanding how high dietary cholesterol influences cholelithogenesis. We explored a relationship between cholesterol absorption efficiency and gallstone prevalence in 15 strains of inbred male mice and the metabolic fate of chylomicron and chylomicron remnant cholesterol in gallstone-susceptible C57L and gallstone-resistant AKR mice. Our results show a positive and significant (P<0.0001, r=0.87) correlation between percent cholesterol absorption and gallstone prevalence rates. Compared with AKR mice, C57L mice displayed significantly greater absorption of cholesterol from the small intestine, more rapid plasma clearance of chylomicrons and chylomicron remnants, higher activities of lipoprotein lipase and hepatic lipase, greater hepatic uptake of chylomicron remnants, and faster secretion of chylomicron remnant cholesterol from plasma into bile. All of these increased susceptibility to cholesterol gallstone formation in C57L mice. We conclude that genetic variations in cholesterol absorption efficiency are associated with cholesterol gallstone formation in inbred mice and cholesterol absorbed from the intestine provides an important source for biliary hypersecretion. Differential metabolism of the chylomicron remnant cholesterol between C57L and AKR mice clearly plays a crucial role in the formation of lithogenic bile and gallstones. PMID- 15749061 TI - A short history of evidence-based obstetric care. AB - Evidence-based obstetric care is a relatively new concept, which had its origins in the early 1970s when Iain Chalmers and his colleagues in Oxford responded to the statement of Archie Cochrane that much of the evidence underpinning obstetric (and other) practices was flawed. They recognized the importance of the quality of evidence in informing clinical decision making, particularly evidence from randomized trials. This was a shift away from opinion-based obstetrics, which up until then had been the dominant paradigm. Since then, there has been an exponential increase in the number and quality of clinical trials in obstetrics, and with their dissemination through the Cochrane database of systematic reviews, there have been many improvements in obstetric practice, more closely aligning it with sound evidence. PMID- 15749062 TI - The tools and techniques of evidence-based medicine. AB - Evidence-based medicine is the conscientious, explicit and judicious use of the current best evidence in making decisions about the care of individual patients. Along with individual clinical expertise, it is a required core skill for clinical problem solving and it is considered to be a comprehensive component of the medical curricula. This chapter is a general overview of the steps to be followed by clinicians to search, identify and appraise the best-available evidence that could help them to resolve a particular clinical problem. It includes the principles for the identification of a clinical problem and its translation into a question, and the main sources for searching and locating the best-available evidence. References for guidelines designed for appraisal of the methods used in the original papers and for the interpretation of its results are also provided. PMID- 15749063 TI - Large trials or meta-analysis? That is not the question. AB - Selection and evaluation of evidence to identify the most effective treatment modalities is a difficult process. Randomized controlled trials are well accepted as the least biased means of evaluating medical, surgical, screening or preventive manoeuvres. The most commonly used statistical strategy to pool results from trials identified during a systematic review is the meta-analysis. Heterogeneity of trial results should be evaluated, interactions and specific populations identified for the planning of the new large trial. PMID- 15749064 TI - Systematic reviews of diagnostic tests: a guide to methods and application. AB - This chapter describes the basic steps in a systematic review to evaluate test accuracy, and the threats to validity of reviews inherent in each step. First, the problems to be addressed are specified in the form of well-structured questions (Step 1). This is a key step, as all other aspects of the review follow directly from the questions. Second, thorough literature searches are conducted to identify potentially relevant studies that shed light on the questions (Step 2). This is one essential feature that makes a review systematic. Third, the quality of the selected studies is assessed (Step 3). Fourth, the evidence concerning study characteristics and results is summarized, and differences between studies are explored (Step 4). When feasible and appropriate, meta analysis helps in collating results. Finally, inferences and recommendations for practice are generated from interpretation and exploration of clinical relevance of the findings (Step 5). These steps are illustrated using a published review concerning the cervicovaginal fetal fibronectin test. PMID- 15749065 TI - Individual patient data meta-analyses. AB - Systematic reviews help people to make well-informed decisions about health care by bringing together the relevant evidence, appraising it and, where appropriate, combining it in a meta-analysis. The ultimate aim for a systematic review should be to present all relevant data on all participants in all research judged to be eligible for the review. In reviews of the effects of interventions, this ultimate aim would be met through the collection and analyses of the results of all relevant randomized trials. Reviews that use individual patient data achieve this by the central collection and analyses of data on each participant in the trials. It allows the reviewer to overcome many of the problems associated with a reliance on published data alone, and some of the problems associated with using aggregate data supplied by the trialist, and will add to the analyses that can be performed. This chapter discusses the rationale for individual patient data reviews and describes some of their features. PMID- 15749066 TI - Evidence and practice: the magnesium sulphate story. AB - There is now strong evidence from systematic reviews of randomised trials to support the use of magnesium sulphate for the prevention and treatment of eclampsia. Magnesium sulphate more than halves the risk of eclampsia for women with pre-eclampsia (relative risk (RR) 0.41, 95% confidence interval (CI) 0.29 0.58; number needed to treat (NNT) 102 (95% CI 72-173) compared to placebo. For treatment of eclampsia, magnesium sulphate lowers the risk of maternal death (RR 0.59, 95% CI 0.37-0.94) and of recurrence of further fits (RR 0.44, 95% CI 0.34 0.57) compared to diazepam. Magnesium sulphate also reduces the risk of further fits compared to phenytoin (RR 0.31, 95% CI 0.20-0.47) and to lytic cocktail (RR 0.09, 95% CI 0.03-0.24). PMID- 15749067 TI - The evidence base for fetal medicine. AB - This chapter looks at the evidence base for the practice of fetal medicine and surgery. There is very little good-quality research and much activity is based on case reports and series. However, the philosophy of evidence-based practice is now accepted and new randomized trials are being published in the literature (e.g. fetal endoscopic tracheal occlusion for congenital diaphragmatic hernia and laser treatment of the twin-to-twin transfusion syndrome). Many well established techniques would not be suitable for randomized trials but, as new techniques related to established practice are introduced (e.g. middle cerebral artery Doppler assessment), well-designed trials are essential to ensure they are safe, clinically useful and at least as good as standard practice. In this unique area of medicine, where ethical and emotional issues interplay with clinical practice, it is important not to introduce exciting techniques without solid evidence that they are beneficial both in the short and the long term. PMID- 15749068 TI - Evidence-based care of recurrent miscarriage. AB - Between 0.5 and 1.0% of couples experience recurrent pregnancy loss (RPL), which is defined as three or more consecutive miscarriages. Losses are classified as pre-embryonic (<5 weeks), embryonic (5-10 weeks) or fetal (>10 weeks). Genetic abnormalities are responsible for RPL in 2-4% of these couples. Inadequate progesterone production has been proposed a cause of RPL and progesterone is given to prevent miscarriage, despite a lack of supportive evidence. The factor V Leiden and prothrombin G20210A mutations are common inherited thrombophilias also associated with RPL. Antenatal thromboprophylaxis is sometimes recommended although no data exist regarding efficacy. Antiphospholipid syndrome is known to cause RPL and antenatal thromboprophylaxis reduces the risk of miscarriage. Uterine abnormalities might also result in RPL. About 50% of cases of RPL have no identifiable cause. Alloimmune incompatibility has been proposed as a cause for RPL in these women. The concept of alloimmune-related RPL has not been scientifically validated. PMID- 15749069 TI - Evidence-based intrapartum care. AB - Routine care in normal labour may range from supportive care at home to intensive monitoring and multiple interventions in hospital. Good evidence of effectiveness is necessary to justify interventions in the normal process of labour. Inadequate evidence is available to support perineal shaving, routine enemas, starvation in labour and excluding the choice for home births. Evidence supports continuity of care led by midwives, companionship in labour, restricting the use of episiotomy, and active management of the third stage of labour, including routine use of 10 units of oxytocin. Both benefits and risks are associated with routine amniotomy, continuous electronic fetal heart rate monitoring, epidural analgesia, and oxytocin-ergometrine to prevent postpartum haemorrhage. More evidence is needed regarding the emotional consequences of labour interventions, home births, vaginal cleansing, opioid use, the partograph, second-stage labour techniques, misoprostol for primary prevention of postpartum haemorrhage, and strategies to promote evidence-based care in labour. PMID- 15749070 TI - Evidence-based vaginal birth after Caesarean section. AB - Caesarean section rates are rising globally. Whether vaginal birth after Caesarean (VBAC) is safe and under what circumstances is increasingly important. This chapter reviews the literature about the risks of VBAC, patient and management factors that may alter risk, and discusses ongoing research as well as suggestions for improving future research. PMID- 15749071 TI - Evidence-based care of women with a multiple pregnancy. AB - Women with a multiple pregnancy face greater risks for themselves and their infants than women pregnant with one child. Pre-pregnancy care should focus on avoiding multiple pregnancy. Early prenatal care centres on determining chorionicity and screening for fetal anomalies, with later care focusing on the presentation, prediction and management of preterm birth, and intrauterine growth restriction. The optimal timing and mode of birth are the focus of current multicentre, randomised, controlled trials. However, the data from such trials on care for women with a multiple pregnancy are limited. Many areas of care require better-quality information, including when using assisted reproductive techniques, the optimal number of embryos to be transferred, care after the diagnosis of chorionicity, and the benefits of specialised multiple pregnancy clinics. Better-quality information is required to inform clinical practice for women with complications of multiple pregnancy, including monoamniotic twin pregnancy, treatment of twin-to-twin transfusion syndrome, and care following single intrauterine fetal death. PMID- 15749072 TI - Evidence-based neonatal care. AB - Randomized controlled trials were introduced into neonatal care in the 1950s when high inspired oxygen concentrations were discovered to be the cause of an epidemic of blindness in preterm babies due to retinopathy of prematurity. Systematic reviews of many randomized controlled trials were published in an important textbook in 1992, 'Effective Care of the Newborn Infant', which was the starting point for the Neonatal Module of the Cochrane Collaboration. The 171 systematic reviews of interventions in neonatology published in the Cochrane Library provide evidence for neonatal care in many areas of the speciality. Some areas, such as management of respiratory distress with surfactant and assisted ventilation, are well covered by reviews, but others, such as resuscitation at birth and management of jaundice, are much less evidence based. Most of the systematic reviews deal with neonatal care in the developed world, and there are only a few of interest to carers in the developing world. PMID- 15749073 TI - Expression pattern of TG-interacting factor 2 during mouse development. AB - TGIF2 is a close homologue of TGIF that functions as a transcriptional corepressor by interaction with transforming growth factor-beta (TGF-beta) activated Smads. Mutations of TGIF have been found in holoprosencephaly, a genetic disease affecting forebrain and craniofacial developmental. Here we analyzed the expression pattern of TGIF2 during mouse embryogenesis. Expression of TGIF2 transcripts was detected at high level at E12.5 and E15.5 in the nervous system including neopallial cortex, mesencephalon, metencephalon, medulla oblongata, spinal cord, trigeminal ganglion, vestibulocochlear ganglion and dorsal root ganglion. In addition, TGIF2 transcripts could be detected in other tissues including heart, lung, liver, pancreas, kidney, small intestine and nasal cavity. These data indicated that TGIF2 has a wide but well controlled expression pattern during mouse development. PMID- 15749074 TI - Baalc, a marker of mesoderm and muscle. AB - Transcripts of the Brain and Acute Leukemia, Cytoplasmic (BAALC) gene are expressed in human neuroectodermal tissues and in CD34-positive bone marrow cells. High transcript levels occur in leukemic blasts from some patients with acute myeloid leukemia (AML), where high expression is an independent marker of poor prognosis. To gain insight into the hitherto unknown function of BAALC/Baalc, we studied its protein expression in embryonic and adult mouse tissue by immunohistochemical analysis. Baalc protein was mainly expressed in developing and mature muscle cells (cardiac, skeletal, and smooth) beginning on day E9 (heart). Signal was seen in the pre-muscle mesodermal cells of the dermatomyotome regions, and the derivatives of the lateral plate and intermediate mesoderm such as smooth muscle wall of the esophagus, stomach, the gut tube, bronchi, small blood vessels, and urinary bladder. This pattern continued through the late embryonic stages into adulthood. Baalc appeared to localize in the cytoplasm, adjacent to the cell membrane. This is distinctly observed in adult skeletal muscle cells. Baalc co-localized with known muscle-associated proteins but not with neural crest or neuronal markers. Scattered expression in adult bone marrow hematopoietic cells and weak expression in the brain neuropil also occurred. In conclusion, BAALC/Baalc is a marker of the mesodermal lineage, especially muscle. PMID- 15749075 TI - Expression of Stat3 in germ cells of developing and adult mouse ovaries and testes. AB - The Signal transducers and activators of transcription (Stat) family of proteins plays diverse roles during differentiation in many tissues. Stat3 is an essential mammalian gene, critical during embryonic development. In mammals, Stat3 is differentially distributed in the cytoplasm of mature oocytes and in preimplantation embryos suggesting that Stat3 may be involved in determination of polarity. Here, we report that Stat3 protein is expressed in the cytoplasm of oocytes from primordial, primary and secondary follicles in the adult ovary and in developing acrosomes of round spermatids in the adult testis. Stat3 is also expressed in gonocytes, prospermatogonia, oogonia and oocytes of embryonic and neonatal gonads. PMID- 15749077 TI - Expression patterns of the homeotic genes Scr, Antp, Ubx, and abd-A during embryogenesis of the cricket Gryllus bimaculatus. AB - We have studied embryogenesis of the two-spotted cricket Gryllus bimaculatus as an example of a hemimetabolous, intermediate germ insect, which is a phylogenetically basal insect and may retain primitive features. We observed expression patterns of the orthologs of the Drosophila homeotic genes, Sex combs reduced (Scr), Antennapedia (Antp), Ultrabithorax (Ubx) and abdominal-A (abd-A) during embryogenesis and compared the expression patterns of these genes with the more basal thysanuran insect, Thermobia domestica (the firebrat), and the derived higher dipteran insect, Drosophila melanogaster. Although Scr is expressed commonly in the presumptive posterior maxillary and labial segment in all three insects, the thoracic expression domains vary. Antp is expressed similarly in the three thoracic segments, the limbs, and the anterior abdominal region among these three insects. The early Antp expression in the firebrat and cricket obeys a segmental register in all three thoracic segments, while in Drosophila its initial expression appears in parasegments 4 and 6. Ubx is expressed in the metathoracic (T3) and abdominal segments similarly in the three insects, whereas the expression pattern in the T3 leg differs among them. abd-A is expressed in the posterior compartment of the first abdominal segment and the remaining abdominal segments in all three insects, although its posterior border varies among them. PMID- 15749076 TI - Cloning and expression of the large zebrafish protocadherin gene, Fat. AB - The cadherin superfamily members play an important role in mediating cell-cell contact and adhesion (Takeichi, M., 1991. Cadherin cell adhesion receptors as a morphogenetic regulator. Science 251, 1451-1455). A distinct subfamily, neither belonging to the classical or protocadherins includes Fat, the largest member of the cadherin super-family. Fat was originally identified in Drosophila. Subsequently, orthologues of Fat have been described in man (Dunne, J., Hanby, A. M., Poulsom, R., Jones, T. A., Sheer, D., Chin, W. G., Da, S. M., Zhao, Q., Beverley, P. C., Owen, M. J., 1995. Molecular cloning and tissue expression of FAT, the human homologue of the Drosophila fat gene that is located on chromosome 4q34-q35 and encodes a putative adhesion molecule. Genomics 30, 207-223), rat (Ponassi, M., Jacques, T. S., Ciani, L., ffrench, C. C., 1999. Expression of the rat homologue of the Drosophila fat tumour suppressor gene. Mech. Dev. 80, 207 212) and mouse (Cox, B., Hadjantonakis, A. K., Collins, J. E., Magee, A. I., 2000. Cloning and expression throughout mouse development of mfat1, a homologue of the Drosophila tumour suppressor gene fat [In Process Citation]. Dev. Dyn. 217, 233-240). In Drosophila, Fat has been shown to play an important role in both planar cell polarity and cell boundary formation during development. In this study we describe the characterization of zebrafish Fat, the first non-mammalian, vertebrate Fat homologue to be identified. The Fat protein has 64% amino acid identity and 80% similarity to human FAT and an identical domain structure to other vertebrate Fat proteins. During embryogenesis fat mRNA is expressed in the developing brain, specialised epithelial surfaces the notochord, ears, eyes and digestive tract, a pattern similar but distinct to that found in mammals. PMID- 15749078 TI - Exclusive expression of hedgehog in small micromere descendants during early embryogenesis in the sea urchin, Hemicentrotus pulcherrimus. AB - Hedgehog (hh) is a multifunctional extracellular protein, and known as an essential signal molecule in morphogenetic movement in animal embryos. We have cloned, sequenced, and studied dynamic localization of Hphh, a hedgehog homologue of the sea urchin, Hemicentrotus pulcherrimus. The origin of Hphh transcribing cells was also verified during early embryogenesis. The amino acid sequence of Hphh shows high homology to Lvhh, an hh homologue cloned in the sea urchin, Lytechinus variegatus. Reverse transcriptase polymerase chain reaction showed that the transcription of Hphh occurred at and after 19 h post-fertilization (19 hpf) mesenchyme blastula stage until, at least, 69 hpf 4-arm pluteus stage. Whole mount in situ hybridization showed Hphh transcription sites in a few cells at the tip of archenteron in 30 hpf gastrulae. At around 45 hpf 2-arm pluteus stage, the number of Hphh transcribed cells was 8, and unequally split to two groups, 5 cells in left coelomic sac and 3 cells in right coelomic sac. A cell lineage tracing by staining the small micromeres with 5-Bromo-2-deoxyuridine showed that Hphh was transcribed exclusively in all the small micromere descendants and comprised the coelomic sacs in 69 hpf plutei. PMID- 15749079 TI - Expression of Fgf10 and Fgf receptors during development of the embryonic chicken stomach. AB - Fibroblast growth factor 10 (FGF10) is involved in numerous different aspects of embryonic development and especially in active epithelial-mesenchymal interactions during morphogenesis of many organs as a mesenchymal regulator by activating its receptors (FGFR1b and FGFR2b) expressed in the epithelial tissue. FGFR2b is also activated by FGF7 although FGF7 does not bind to FGFR1b. To provide basic data to analyze function of FGFs in the developing gut, here we cloned Fgf7 and studied expression patterns of Fgf7, Fgf10 and Fgfr1-4 during the development of chicken stomach (glandular stomach; proventriculus and muscular stomach; gizzard). Fgf10 is expressed both in the proventricular and gizzard mesenchyme while Fgf7 is expressed only in gizzard mesenchyme. Fgfr1-4 are expressed both in the epithelium and mesenchyme with a different spatial expression patterns. Furthermore, RT-PCR analysis reveals that Fgfr1b and Fgfr2b are expressed only in epithelia of both organs. PMID- 15749080 TI - Expression of MUK/DLK/ZPK, an activator of the JNK pathway, in the nervous systems of the developing mouse embryo. AB - C-Jun N-terminal kinase (JNK) is implicated in regulating the various cellular events during neural development that include differentiation, apoptosis and migration. MUK/DLK/ZPK is a MAP kinase kinase kinase (MAPKKK) enzyme that activates JNK via MAP kinase kinases (MAPKK) such as MKK7. We show here that the expression of MUK/DLK/ZPK protein in the developing mouse embryo is almost totally specific for the neural tissues, including central, peripheral, and autonomic nervous systems. The only obvious exception is the liver, in which the protein is temporally expressed at around E11. The expression becomes obvious in the neurons of the brain and neural crest tissues at embryonic day 10 (E10) after neuron production is initiated. By E14, MUK/DLK/ZPK proteins are found in various neural tissues including the brain, spinal cord, sensory ganglia (such as trigeminal and dorsal root ganglia), and the sympathetic and visceral nerves. The localization of MUK/DLK/ZPK protein in neural cells almost consistently overlapped that of betaIII-tubulin, a neuron specific tubulin isoform, and both proteins were more concentrated in axons than in cell bodies and dendrites. The intensely overlapping localization of betaIII-tubulin and MUK/DLK/ZPK indicated that this protein kinase is tightly associated with the microtubules of neurons. PMID- 15749081 TI - Asymmetric expression of Syndecan-2 in early chick embryogenesis. AB - Chicken Syndecan-2 (cSyndecan-2) is the homologue of Xenopus Syndecan-2, a member of the heparan sulfate proteoglycan family with an important role in left-right patterning in frog embryos. A relationship to LR asymmetry in other species has not been reported. We show that cSyndecan-2 is expressed throughout the primitive streak between st. 1 and 3 in the chick embryo, and is restricted to the rostral and caudal tips of the primitive streak at st. 4. It displays distinct left-right asymmetry, being expressed in the right side of Hensen's node at st. 5. The asymmetric expression of cSyndecan-2 is maintained around the node between st. 5 and 7. At early somite stages, somites and neural folds express cSyndecan-2. The somite expression disappears by st. 11, but strong expression in the neural tube continues. Our data reveal a new asymmetric transcript in the chick embryo and indicate that in contrast to protein-level asymmetries, which underlie syndecan-2 function in Xenopus, chick syndecan-2 exhibits asymmetry at the mRNA level. PMID- 15749082 TI - Asymmetric expression of Gli transcription factors in Hensen's node. AB - Avian left-right (L/R) axis determination involves the establishment of asymmetric gene expression in Hensen's node, resulting in two discrete signalling pathways on the left and right sides of the embryo. The extracellular signalling molecule Sonic Hedgehog (SHH) is known to be an important left-side determinant. Transcription of Shh is initially bilateral in Hensen's node (stage 4), but is restricted to the left side by stage 5. The Gli genes (Gli1, Gli2 and Gli3) are the main transcriptional mediators of the Hedgehog pathway in vertebrates. GLI1 and GLI2 are primarily transcriptional activators of Hedgehog target genes, while GLI3 is primarily a transcriptional repressor of Hedgehog targets. In order to gain insight into the mechanisms of asymmetrical Hedgehog signal transduction in the node, we have analysed the expression patterns of the Gli genes in Hensen's node from stage 4 to stage 8. Here, we reveal that the Gli genes are asymmetrically expressed in Hensen's node: Gli1 and Gli2, are expressed on the left side, while Gli3 is expressed on the right side. PMID- 15749084 TI - Regulated expression pattern of gremlin during zebrafish development. AB - Xenopus laevis Gremlin has been isolated as a novel dorsalizing factor, belonging to a family of secreted proteins with axial patterning activity . In a search for genes that control development in zebrafish (Danio rerio), we have identified a sequence homologous to Xenopus gremlin. This paper describes the cloning of zebrafish gremlin (grm) and its expression pattern during development. Our results show that grm encodes a maternal transcript, and the zygotic transcription is turned on at the mid-blastula transition (MBT), when grm is detected in the entire blastoderm. In the gastrula grm becomes restricted to the dorsolateral region of the embryo, and during somitogenesis it is strongly expressed in the presomitic mesoderm and developing somites, and in the ventral neural tube. From 24 hpf to 48 hpf, we show that grm transcription is downregulated in the whole embryo, even though Grm protein is still present and localized into the entire myotome at 48-72 hpf. Finally, grm transcript is strongly downregulated in fibroblast growth factor-8 (fgf8) and sonic hedgehog (shh) mutants, thus implicating a putative role of Fgf/Shh signalling loop in grm expression regulation. PMID- 15749083 TI - Aortic carboxypeptidase-like protein is expressed in collagen-rich tissues during mouse embryonic development. AB - Aortic carboxypeptidase-like protein (ACLP) was originally identified in vascular smooth muscle cells and contains discoidin and catalytically inactive metallocarboxypeptidase domains. ACLP is a secreted protein that associates with the extracellular matrix and is essential for abdominal wall development and contributes to dermal wound healing. Because of these developmental and adult phenotypes, we examined the expression of ACLP by immunohistochemistry throughout mouse embryonic development. ACLP was not detected in 7.5 days post-coitum (dpc) embryos, however at 9.5 dpc low levels of expression were detected in the somites and dorsal aorta. Expression was detected in both the yolk sac and embryonic vasculature at 10.5d pc. ACLP expression increased in both large and small blood vessels at 11.5 and 13.5 dpc and intense expression was detected within the vascular smooth muscle layer in 16.5 dpc embryos. At later developmental time points, discrete areas of ACLP expression were detected in the mesenchymal cells in the dermal layer, developing skeletal structures, connective tissue, and in the umbilical ring and vessels. The predominance of ACLP immunoreactivity localized with collagen-rich regions including tendons and basement membranes. Overall, the developmental expression pattern is consistent with a regulatory or structural role in the abdominal wall, vasculature, and dermis. PMID- 15749085 TI - Spatial and temporal distribution of the traf4 genes during zebrafish development. AB - The tumor necrosis factor-associated factor 4 (TRAF4) is a particular member of the TRAF protein family since it is not involved in the Tumor Necrosis Factor (TNF) and Interleukin-1 (IL-1) signaling pathways. In the present study, we cloned two zebrafish orthologs of the human traf4, traf4a and traf4b, which are the first TRAFs described in zebrafish. During embryogenesis, traf4b expression is present in a weak ubiquitous manner. In contrast, traf4a exhibits a highly specific expression pattern in the sensorial and neural cells, and the somites of embryos. This gene is tightly regulated during embryogenesis. Together, our data show that traf4 is conserved during evolution, and traf4a is the zebrafish ortholog of traf4. PMID- 15749086 TI - Gene expression pattern of Claudin-1 during chick embryogenesis. AB - Claudins are a family of proteins that are localized to tight junctions at the apical surface of epithelial cell layers. Over 24 family members have been identified in vertebrates. Despite being well-studied with respect to their function in tight junction selectivity and permeability, the embryonic expression patterns of most claudin family members have not been thoroughly investigated. Here, we report the cloning and expression pattern of a novel chick claudin family member that is most closely related to human claudin-1. Chick claudin-1 was expressed throughout the ectoderm of stage 4-6 chick embryos. Claudin-1 expression was particularly high in the neural epithelium and open neural tube, but decreased as the neural tube closed. High levels of claudin-1 expression were also observed in the developing otic vesicle, nasal placode, ectodermal component of the pharyngeal arches, and in the apical ectodermal ridge of the limb bud from stage 17 onwards. Claudin-1 expression was also detected in scleral papillae, feather buds and migrating primordial germ cells. Lower levels of claudin-1 expression were observed in the endoderm, the ventral pharynx, and several of its derivatives including the bronchi, developing lung epithelium, esophagus, and gut. Claudin-1 expression was detected in the nephric duct and the mesonephros, which are epithelialized derivatives of the intermediate mesoderm, but not in any other mesodermal derivates, including the heart, somites and developing muscle. With the exception of the migrating primordial germ cells and the primitive streak, all other tissues that expressed significant levels of claudin-1 were epithelialized. PMID- 15749087 TI - Expression pattern of NOGO and NgR genes during human development. AB - Nogo protein has been identified as the component of central nervous system (CNS) myelin that limits axonal regeneration. We investigated the expression of the genes encoding Nogo and its receptor, NgR, between weeks eight and 23 of human embryonic development, by quantitative radioactive in situ hybridization. At 8 weeks, we detected NOGO and NgR transcripts in developing neuronal and non neuronal structures. We focused on two different structures: the brain and the dental germs. During this period of development, NOGO and NgR transcripts colocalized in the cortical and ventricular zones of the brain, with expression strongest for these two genes in the postmitotic cells of the cortical plate. In developing dental germs, NgR was more strongly expressed than NOGO at 16 and 21 weeks. NOGO and NgR were expressed in zones of epithelium-mesenchyme interaction, which induce the differentiation of ameloblasts/odontoblasts. These genes were expressed most strongly in differentiated cells. PMID- 15749088 TI - Expression of fibroblast growth factors 18 and 23 during human embryonic and fetal development. AB - Fibroblast Growth Factor (FGF) 18 and 23 are two recently identified members of the FGF family, a family of structurally related polypeptides with diverse roles in physiological and pathological processes. Studies mostly performed in rodents and chicken have demonstrated that FGF18 is a pleiotropic growth factor involved in the development of various organs, while there are no data supporting a direct role of FGF23 in cell proliferation or differentiation either in physiology or pathology in any species. However, it is now established that FGF23 can be a humoral messenger and an important regulator of phosphate homeostasis and vitamin D metabolism. As a first step towards elucidating the roles of these FGF in human development, we examined FGF18 and FGF23 mRNA expression by in situ hybridization in whole human embryos at 30 days and 8 weeks of gestation (GW) and in specific fetal tissues at different ages. We report a highly restricted expression pattern for both FGF genes in human embryonic development. PMID- 15749089 TI - Design of a portable urine glucose monitoring system for health care. AB - This paper describes the design of a monitoring system that can be used to measure urine glucose during daily life. It consists of a bio-chemical sensor, hardware with PIC microcontroller and control circuits, and signal analyzing part. To evaluate the performance, we compared the analyzed glucose levels of the developed system to a standard instrument, YSI glucose analyzer, based on regression analysis using standard glucose solutions mixed with urine. Also, standard deviation and coefficient of variation were computed. In conclusion, the developed system showed it could be used for the measurement of urine glucose. PMID- 15749090 TI - Thermoregulatory model for prediction of long-term cold exposure. AB - A multi-segmental mathematical model has been developed for predicting shivering and thermoregulatory responses during long-term cold exposure. The present model incorporates new knowledge on shivering thermogenesis, including the control and maximal limits of its intensity, inhibition due to a low core temperature, and prediction of endurance time. The model also takes into account individual characteristics of age, height, weight, % body fat, and maximum aerobic capacity. The model was validated against three different cold conditions i.e. water immersion up to 38 h and air exposure. The predictions were found to be in good agreement with the observations. PMID- 15749091 TI - Changes in the slope of the first major deflection of the ECG complex during acute coronary occlusion. AB - To evaluate the effect of acute coronary occlusion (ACO) during percutaneous coronary intervention (PCI) on the slope of the first major deflection of the QRS complex (the initial QRS slope). Standard ECG signals of 18 patients (89 leads), undergoing PCI were recorded prior to and during ACO. The initial QRS slope was calculated in the baseline state and during ACO. Changes in the standard ECG were detected in 36 of 89 leads (40%). The initial QRS slope during ACO was significantly different from baseline in 74 of 89 leads (83%). The specificity of the change in the slope during ACO was low (29%). PMID- 15749092 TI - Predicting survival time for kidney dialysis patients: a data mining approach. AB - The cost for providing care for patients on hemodialysis due to end stage kidney disease is high. Finding ways to improve patient outcomes and reduce the cost of dialysis is important. Dialysis care is intricate and multiple factors may influence patient survival. Over 50 parameters may be monitored on a regular basis in providing kidney dialysis treatments. Understanding the collective role of these parameters in determining outcomes for an individual patient and administering individualized treatments allowing specific interventions is a challenge. Individual patient survival may depend on a complex interrelationship between multiple demographic and clinical parameters, medications, medical interventions, and the dialysis treatment prescription. In this research, data preprocessing, data transformations, and a data mining approach are used to elicit knowledge about the interaction between many of these measured parameters and patient survival. Two different data mining algorithms were employed for extracting knowledge in the form of decision rules. These rules were used by a decision-making algorithm, which predicts survival of new unseen patients. Important parameters identified by data mining are interpreted for their medical significance. The concepts introduced in this research have been applied and tested using data collected at four dialysis sites. The computational results are reported in the paper. PMID- 15749093 TI - Interactive 3D segmentation using connected orthogonal contours. AB - This paper describes a new method for interactive segmentation that is based on cross-sectional design and 3D modelling. The method represents a 3D model by a set of connected contours that are planar and orthogonal. Planar contours overlayed on image data are easily manipulated and linked contours reduce the amount of user interaction.1 This method solves the contour-to-contour correspondence problem and can capture extrema of objects in a more flexible way than manual segmentation of a stack of 2D images. The resulting 3D model is guaranteed to be free of geometric and topological errors. We show that manual segmentation using connected orthogonal contours has great advantages over conventional manual segmentation. Furthermore, the method provides effective feedback and control for creating an initial model for, and control and steering of, (semi-)automatic segmentation methods. PMID- 15749094 TI - Three-dimensional hip morphology analysis using CT transverse sections to automate diagnoses and surgery managements. AB - This paper describes an image analysis method that evaluates bone morphology of hip structures including the femur stem, trochanter, neck and head, acetabulum, and pelvis to automate hip diagnoses and surgical managements. On every CT transverse section, radial B-spline curves are used to approximate the ellipse like acetabulum and femur head and stem. The femur neck is approximated as trapezoid-like and the pelvis horizontally symmetrical structure. The centers of the ellipse-like structures from transverse sections are used to determine 3D axes of the femur stem, head, and acetabulum. The centerlines of the neck or the pelvis on the sections are used to determine the neck axis or pelvis centerplane. Boundary changes of these structures are recognized as concave, convex and hole features that are then identified as fractures, tumors, and spurs. Based on the geometric evaluations of these structures and features, hip surgeries including tumor dissect and bone graft, open reduction using plates, screws and nails, and arthroplasty are automatically managed to achieve the normal hip function including dissection of tumors and reduction of dislocations and angular deviations between the hip structures. This prototype system can be used as a qualitative and quantitative tool for the diagnosis of various hip diseases and for the planning of accurate surgical procedures. A series of examples and four case studies illustrate this automated method can be used to accurately diagnose hip diseases and manage hip surgeries, and train operators. PMID- 15749095 TI - Non-poly-DNA viruses, their parasitic wasps, and hosts. PMID- 15749096 TI - Characteristics of pathogenic and mutualistic relationships of ascoviruses in field populations of parasitoid wasps. AB - Ascoviruses are disseminated among larvae in lepidopteran populations by parasitic wasps during oviposition. Ascovirus relationships with these wasps vary from pathogenic to mutualistic, and experimentally can be shown possibly to be commensal non-pathogenic virus having little or no effect. Most ascoviruses are pathogens that female wasps vector mechanically. Other ascoviruses have a more intimate relationship with their wasp vectors in that their genome is stably maintained in all wasp nuclei through several generations by vertical transmission. In this relationship, these viruses are mutualistic, enhancing the successful development of the wasp larvae by suppressing lepidopteran defence mechanisms. The DpAV4 ascovirus is a mutualist in certain Diadromus wasps but is pathogenic or not when vectored by other species of this genus. These various biologies suggest that ascovirus/wasp relationships depend on wasp regulatory factors that control virus replication. Thus, certain ascoviruses can potentially have either a pathogenic, mutualistic, or non-pathogenic relationship with a specific wasp vector, the type of relationship being dependent upon the species system in which the relationship evolved. Finally, because ascoviruses appear to be related to ichnoviruses (Polydnaviridae), the DpAV4/Diadromus system constitutes a possible interesting intermediate between the pathogenic ascoviruses and symbiotic viruses that evolved to be ichnoviruses. PMID- 15749097 TI - Phenoloxidase-like activities and the function of virus-like particles in ovaries of the parthenogenetic parasitoid Venturia canescens. AB - The ichneumonid endoparasitoid Venturia canescens successfully develops inside the hemocoel of another insect by using maternal protein secretions, including nucleic acid-free virus-like particles (VLPs), to manipulate host physiology. These VLPs consist of four major proteins, which are produced mainly in the calyx tissue and transferred into the host insect together with the egg. One of the protein-coding genes (vlp1), with similarities to phospholipid-hydroperoxide glutathione peroxidases (PHGPx), exists in allelic forms producing two protein variants with different protein properties. Here, we summarise observations indicating that oocytes and eggs are the source of reactive electrons, which potentially damage the lining and membranes of calyx tissues. We discuss the possible role of VLP1 in counteracting the damaging effects of oxidised phospholipids on membranes surrounding VLPs in the calyx lumen. PMID- 15749098 TI - Presence of a novel small RNA-containing virus in a laboratory culture of the endoparasitic wasp Venturia canescens (Hymenoptera: Ichneumonidae). AB - Parasitoid wasps use a variety of mechanisms to alter their host's physiology to the benefit of the developing endoparasite inside the host larva. Association of certain wasps with viruses and virus-like particles (VLPs) that contribute to their success in parasitism is one of the fascinating evolutionary adaptations conferring active or passive protection for the endoparasite from the host immune system. Venturia canescens has been shown to produce VLPs that provide protection for the developing parasitoid egg inside the host, Ephestia kuehniella. Here, we report on the presence of a novel small RNA-containing virus from V. canescens, designated as VcSRV, occurring in the ovaries of the wasp. The virus particles are found together with VcVLPs in the lumen of the calyx region of the ovaries and are injected together with the egg and VcVLPs into E. kuehniella larvae where they enter hemocytes. Alignment of the RNA-dependent RNA polymerase gene of VcSRV indicates that the virus most likely belongs to the recently described genus Iflavirus. PMID- 15749099 TI - Commensal and mutualistic relationships of reoviruses with their parasitoid wasp hosts. AB - During evolution, certain endoparasitoid wasps have developed mechanisms to suppress the defence systems of their hosts. For this purpose, these species, all of which belong to the families Ichneumonidae and Braconidae, inject various kinds of virus-like particles. The most studied of these particles are classified as polydnaviruses (family Polydnaviridae) which are symbiotic viruses. Over the past decade, it has also been shown that several wasp species harbour reoviruses (family Reoviridae), and that two of these suppress host defence, allowing the development of the parasitoid eggs. In this paper, we summarize the key features of these viruses and their relationships with their wasp hosts. Five reoviruses are known that appear to be non-pathogenic for the wasps. Three of these, McRVLP, HeRV, OpRVLP, use their wasp hosts as vectors, and do not appear to be involved in host defence suppression. The fourth, DpRV-1, is a commensal reovirus detected in most field populations of the wasp, Diadromus pulchellus. This reovirus is always found associated with an ascovirus, DpAV-4a, which is indispensable for host immune suppression. Although DpRV-1 has not been shown to directly increase D. pulchellus parasitic success, it may contribute to this success by retarding DpAV-4a replication in the wasp. The fifth reovirus, DpRV-2, occurs in a specific population of D. pulchellus in which DpRV-1 and DpAV-4 are absent. This virus has a mutualistic relationship with its wasp host, as its injection by females during oviposition is essential for host immunosuppression. Interestingly, these viruses belong to several different reovirus genera. PMID- 15749100 TI - Localization of intracellular calcium release in cells injured by venom from the ectoparasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) and dependence of calcium mobilization on G-protein activation. AB - Venom from the ectoparasitic wasp Nasonia vitripennis induces cellular injury that appears to involve the release of intracellular calcium stores via the activation of phospholipase C, and culminates in oncotic death. A linkage between release of intracellular Ca2+ and oncosis has not been clearly established and was the focus of this study. When BTI-TN-5B1-4 cells were treated with suramin, an uncoupler of G-proteins, venom-induced swelling and oncotic death were inhibited in a dose-dependent manner for at least 24 h. Suramin also blocked increases in free cytosolic [Ca2+], arguing that venom induces calcium mobilization through G-protein signaling pathways. Endoplasmic reticulum (ER) was predicted to be the source of intracellular calcium release, but labeling with the fluorescent probe ER-tracker revealed no indication of organelle swelling or loss of membrane integrity as would be expected if the Ca(2+)-ATPase pump was disabled by crude venom. Incubation of cell monolayers with calmodulin or nitrendipine, modulators of ER calcium release channels, neither attenuated nor augmented the effects of wasp venom. These results suggest that wasp venom stimulates calcium release from ER compartments distinct from RyRs, L-type Ca2+ channels, and the Ca(2+)-ATPase pump, or calcium is released from some other intracellular store. A reduction of mitochondrial membrane potential delta psi(m) appeared to precede a rise in cytosolic free Ca2+ as evidenced by fluorescent microscopy using the calcium-sensitive probe fluo-4 AM. This argues that the initial insult to the cell resulting from venom elicits a rapid loss of (delta psi(m)), followed by unregulated calcium efflux from mitochondria into the cytosol. Mobilization of calcium in this fashion could stimulate cAMP formation, and subsequently promote calcium release from NAADP-sensitive stores. PMID- 15749102 TI - Asobara, braconid parasitoids of Drosophila larvae: unusual strategies to avoid encapsulation without VLPs. AB - Ichneumonoidae parasitoids have been well described for their regulatory effects on host physiology which are usually associated with the activity of polydnaviruses (PDVs) or viruslike-particles (VLPs) injected by the female wasps at oviposition. Among them, parasitoids of the braconid families display specific characteristics like the required activity of secretions from the maternal venom glands or of teratocytes from embryological origin. However, none of these features were observed in two braconid species of the Asobara genus parasitizing Drosophila hosts. In the absence of PDVs and VLPs, the two species A. tabida and A. citri seem to have developed unique strategies to avoid immunity defenses and to succeed in their Drosophila larval hosts. The aim of this study is to report on the complex relationships of braconid parasitoids with their hosts and to present some of the insights from studying Drosophila parasitoids. PMID- 15749101 TI - Haemocyte changes in D. Melanogaster in response to long gland components of the parasitoid wasp Leptopilina boulardi: a Rho-GAP protein as an important factor. AB - The hymenopteran wasp Leptopilina boulardi (Figitidae) is a larval solitary parasitoid of Drosophila larvae of the melanogaster sub-group. The factors used by parasitoid females to prevent encapsulation of their eggs by the host are localized in the female long gland and reservoir. We report here the physiological effects of these factors on host haemocytes using in vivo injection experiments. The total number of haemocytes, the number of plasmatocytes and the number of crystal cells were not modified by injection of long gland extracts. In contrast, long gland extracts either from virulent or avirulent strains had a significant effect on the lamellocyte number. Compared to the Ringer control, the avirulent long gland products induced an increase of the lamellocyte number while virulent extracts induced a drastic decrease together with an alteration of the morphology of these cells. Interestingly, changes in the lamellocyte morphology were also observed following injection of the P4 protein, a major component of L. boulardi female long glands that displays a strong immune suppressive effect on Drosophila larvae. The implication of the P4 protein in suppressing the host cellular immunity is discussed in correlation with its predicted molecular function as a Rho-GAP protein. PMID- 15749103 TI - Biogenesis, structure, and immune-suppressive effects of virus-like particles of a Drosophila parasitoid, Leptopilina victoriae. AB - Drosophila melanogaster larvae are attacked by virulent strains of parasitoid wasps. Females of Leptopilina heterotoma produce virus-like particles (VLPs) that efficiently destroy lamellocytes, a major larval immune effector cell type. We report here that L. victoriae, a closely related wasp species, also produces VLPs that trigger immune suppression responses in fly hosts. We compare the ability of immune suppression of the two parasitoids using a mutant host strain hopscotch(Tumorous-lethal) (hop(Tum-l)). hop(Tum-l) larvae have two defects of hematopoietic origin: overproliferation of hemocytes and constitutive encapsulation of self-tissue by lamellocytes. The encapsulation phenotype is suppressed weakly by L. victoriae and strongly by L. heterotoma. In vitro studies on hop(Tum-l) lamellocytes show that VLP-containing fluid from either wasp species induces lamellocyte lysis, but with different kinetics. Previously undocumented precursors of L. victoriae VLPs are synthesized in the long gland and are first visible within canals connecting secretory cells to the long gland lumen. VLP assembly occurs in the lumen. VLPs show multiple electron-dense projections surrounding a central core. Maturing particles appear segmented, singly or in arrays, embedded in the reservoir matrix. In sections, mature particles are pentagonal or hexagonal; the polygon vertices extending into spikes. Our results suggest that L. victoriae is likely to promote immune suppression by an active mechanism that is mediated by VLPs, similar to that used by L. heterotoma. PMID- 15749104 TI - Drosophila serpin 27A is a likely target for immune suppression of the blood cell mediated melanotic encapsulation response. AB - Avirulent strains of the endoparasitoid Leptopilina boulardi succumb to a blood cell-mediated melanotic encapsulation response in host larvae of Drosophila melanogaster. Virulent wasp strains effectively abrogate the cellular response with substances introduced into the host that specifically target and effectively suppress one or more immune signaling pathways, including elements that control phenoloxidase-mediated melanotic encapsulation. The present study implicates involvement of the Drosophila Toll pathway in cellular innate immunity by regulating the serine protease inhibitor Serpin 27A (Spn27A), which normally functions as a negative regulator of phenoloxidase. The introduction of Spn27A into normally highly immune competent D. melanogaster larvae significantly reduced their ability to form melanotic capsules around eggs of L. boulardi. This study confirms the role of Spn27A in the melanization cascade and establishes that this pathway and associated blood cell responses can be activated by parasitization. The activation of phenoloxidase and the site-specific localization of the ensuing melanotic response are such critical components of the blood cell response that Spn27A and the signaling elements mediating its activity are likely to represent prime targets for immune suppression by L. boulardi. PMID- 15749105 TI - Comparative analysis of selected genes from Diachasmimorpha longicaudata entomopoxvirus and other poxviruses. AB - The Diachasmimorpha longicaudata entomopoxvirus (DlEPV) is the first symbiotic EPV described from a parasitic wasp. The DlEPV is introduced into the tephritid fruit fly larval host along with the wasp egg at oviposition. We sequenced a shotgun genomic library of the DlEPV DNA and analyzed and compared the predicted protein sequences of eight ORFs with those of selected poxviruses and other organisms. BlastP searches showed that five of these are homologous to poxvirus putative proteins such as metalloprotease, a putative membrane protein, late transcription factor-3, virion surface protein, and poly (A) polymerase (PAP) regulatory small subunit. Three of these are similar to those of other organisms such as the gamma-glutamyltransferase (GGT) of Arabidopsis thaliana, eukaryotic initiation factor 4A (eIF4A) of Caenorhabditis briggsae and lambda phage integrase (lambda-Int) of Enterococcus faecium. Transcription motifs for early (TGA,A/T,XXXXA) or late (TAAATG, TAAT, or TAAAT) gene expression conserved in poxviruses were identified with those ORFs. Phylogenetic analysis of multiple alignments of five ORFs and 20 poxvirus homologous sequences and of a concatenate of multiple alignments suggested that DlEPV probably diverged from the ancestral node between the fowlpox virus and the genus B, lepidopteran and orthopteran EPVs, to which Amsacta moorei and Melanoplus sanguinipes EPV, respectively, belong. The DlEPV putative GGT, eIF4A, and lambda-Int contained many conserved domains that typified these proteins. These homologues may be involved in either viral pathogenicity or enhancing parasitism via the gamma-glutamyl cycle and compensation of eIF4A levels in the parasitized fly, or via the integration of a portion of the viral genome into the wasp and/or parasitized fly. PMID- 15749106 TI - Morphogenesis and cytopathic effects of the Diachasmimorpha longicaudata entomopoxvirus in host haemocytes. AB - The Diachasmimorpha longicaudata entomopoxvirus (DlEPV), the first reported symbiotic entomopoxvirus, occurs in the venom apparatus of D. longicaudata female wasps and is introduced into Anastrepha suspensa larvae during parasitism. The DlEPV 250-300 kb double stranded DNA genome encodes putative proteins having 30 to >60% amino acid identity with poxvirus homologs such as DNA helicase, DNA dependent RNA polymerase, and the poxvirus-specific rifampicin resistance protein. Although the molecular characterization of DlEPV is progressing, little is known about its morphogenesis in and effects on host haemocytes. This paper describes (1) haemocytes of third instar A. suspensa, (2) DlEPV infection and morphogenesis, and (3) DlEPV-induced changes in haemocytes. A. suspensa third instars have 3-4 haemocyte morphotypes. Dot blots of DNA from infected haemocytes hybridized with a digoxigenin-labeled DlEPV genomic probe as early as 4 h post parasitism (hpp) and the intensity of the signal increased with time through 40 hpp. Immunofluorescence microscopy localized DlEPV proteins in cytoplasmic (but not nuclear) sites of infected haemocytes, within 24-36 hpp. Electron microscopy confirmed the presence of viral envelopes, immature spheroids with centric nucleoids, budding virus, and extracellular enveloped virus in three haemocyte types, 24-84 hpp and later. Infected haemocytes exhibited blebbing, DNA concatenation, and inability to encapsulate sephadex beads in vitro. These data indicate that DlEPV disrupts the normal function of host haemocytes, thereby insuring the successful development of D. longicaudata offspring and as such should be regarded as a symbiont of the wasp. PMID- 15749107 TI - Transmission of the Diachasmimorpha longicaudata rhabdovirus (DlRhV) to wasp offspring: an ultrastructural analysis. AB - During oviposition, the parasitic wasp Diachasmimorpha longicaudata introduces an entomopoxvirus (DlEPV) and a rhabdovirus (DlRhV) into larvae of its tephritid fruit fly host Anastrepha suspensa. DlEPV and DlRhV replicate, respectively, in host hemocytes and epidermal cells. Both viruses, like many beneficial viruses of parasitic wasps, are retained in all wasp generations but their avenue(s) of transmission are unknown. This study tests the hypothesis that DlRhV is transmitted transovarially or through larval feeding on infected host hemolymph. Transmission electron microscopy (TEM) revealed no virions in pre-vitellogenic or vitellogenic ova, or in the lateral oviduct of D. longicaudata females. However, numerous virions occurred in subchorionic regions of 33-36-h-old oviposited eggs. This suggests that DlRhV is introduced into the egg either as (a) intact virions after chorionogenesis but prior to oviposition and/or as (b) unencapsidated RNA molecules, undetectable by TEM in pre-vitellogenic ova, that subsequently replicate and assemble into mature virions. DlRhV particles also occurred in the midgut lumen of 20-24-h-old wasp first instars, suggesting that they were ingested. These virions may have been released from the egg into the hemolymph during hatching or may have come from virions introduced by the female wasp directly into the host, separate from the egg. DlRhV particles were also evident in the intracellular vesicles and intercellular spaces of the larval midgut. Taken together, these data support the hypothesis that DlRhV is transovarially transmitted as virions and/or as unencapsidated RNA. Further studies are needed to determine whether the DlRhV that ultimately resides within the female wasp's accessory gland filaments is the progeny of the virus from the egg and/or larval midgut cells. PMID- 15749108 TI - The spatial resolutions of the apposition compound eye and its neuro-sensory feature detectors: observation versus theory. AB - For 100 years three ideas dominated efforts to understand the apposition compound eye. In Muller's theory, the eye viewed the panorama through an array of little windows without overlaps and without gaps, with no details within windows. Spatial resolution then depended on the interommatidial angle (Deltaphi) and the number of ommatidia. In the second proposal, the insect detected the temporal modulation of the light, which was limited by the aperture of the lens and the wavelength, assuming good focus. Modulation is the change of intensity in the receptor, usually caused by motion of a spatial contrast in the stimulus. Thirdly, motion was detected from the successive temporal modulations at adjacent visual axes. Recently, two more principles arose. The light-sensitive elements, called rhabdomeres, project through the nodal point of the lens to the outside world, and the resolution was limited by their grain size, like the pixels in a digital camera. Finally, detection of contrast and colour was limited by the signal/noise ratio (SNR) which was improved by brighter light and more visual pigment. These five physical principles provide satisfying explanations of eye function but they all originated from theory. Actual measurements of resolution depend on the operation of the test. The visual system of the honeybee recognizes a limited variety of simple cues, but there is no evidence that the pattern of ommatidial stimulation is re-assembled, or even seen. The known cues are: the temporal modulation of groups of receptors, the direction and angular velocity of motion, some measure of the spatial disruption of the pattern or the length of edge (related to spatial frequency and contrast), colour, the intensity, the position of the centre and the size of large well-separated areas of black or colour, the angle of orientation of a bar or grating, radial or tangential edges, and bilateral symmetry. Neurons connected to more than two adjacent ommatidia collaborate in the detection of cues, and the resolution depends on the neuro sensory feature detectors at work at the time. Although some behavioural and electrophysiological measurements give a spatial resolution similar to the interommatidial angle, different spatial properties of neuro-sensory detectors predominate at different light intensities and with a diurnal rhythm. During the long history of this topic, the belief that the resolution ought to be Deltaphi has frequently been overturned by experimental measurement. PMID- 15749109 TI - Diuretic factors and second messengers stimulate secretion of the organic cation TEA by the Malpighian tubules of Drosophila melanogaster. AB - This study showed that four factors which stimulate transepithelial fluid secretion and inorganic ion transport across the main segment of the Malpighian tubules of Drosophila melanogaster also stimulate transepithelial secretion of the prototypical organic cation tetraethylammonium (TEA). TEA fluxes across the Malpighian tubules and gut were measured using a TEA-selective self-referencing (TEA-SeR) microelectrode. TEA flux across isolated Malpighian tubules was also measured using a TEA-selective microelectrode positioned in droplets of fluid secreted by tubules set up in a modified Ramsay assay. TEA flux was stimulated by the intracellular second messengers cAMP and cGMP, which increase the lumen positive transepithelial potential (TEP), and also by tyramine and leucokinin-I (LK-I), which decrease TEP. The largest increase was measured in response to 1 micromol l-1 LK-I which increased transepithelial TEA flux by 72%. TEA flux in the lower tubule was stimulated slightly (13%) by 1 micromol l-1 tyramine but not by any of the other factors. TEA flux across the midgut was unaffected by cAMP, cGMP or tyramine. This is the first study to demonstrate the effects of insect diuretic factors and second messengers on excretion of organic cations. PMID- 15749110 TI - Circadian mating activity and effect of pheromone pre-exposure on pheromone response rhythms in the moth Spodoptera littoralis. AB - Mating in moths is generally mediated by female-produced sex pheromones. Mating activity, female pheromone production/release and male pheromone responsiveness all show diurnal variations in many species. We found that the response of the male Egyptian cotton leafworm, Spodoptera littoralis, to sex pheromone gland extracts showed a diel rhythm in olfactometer tests, and the variation was persistent for at least 1 day in constant darkness. High male response to sex pheromone was correlated in time with high mating and locomotor activity. Male S. littoralis, maintained in constant darkness and exposed to pheromone gland extracts on a daily basis, showed an induced temporal variation in response after several days, in contrast to unexposed males. This suggests that in the absence of other external zeitgebers, exposure to sex pheromone may function to synchronise circadian behavioural rhythms in male moths. The daily rhythm in mating activity in S. littoralis is also shown to be persistent for at least 2 days in constant darkness. Pairs mated significantly less when either the male or female had been raised in a light:dark cycle 10 h out of phase, indicating that the proposed circadian rhythm in mating activity is composed of rhythmic mating preference/ability in both sexes. PMID- 15749111 TI - Stage-dependent strategies of host invasion in the egg-larval parasitoid Chelonus inanitus. AB - Chelonus inanitus (Braconidae) is a solitary egg-larval parasitoid which lays its eggs into eggs of Spodoptera littoralis (Noctuidae); the parasitoid larva then develops in the haemocoel of the host larva. Host embryonic development lasts approx. 3.5 days while parasitoid embryonic development lasts approx. 16 h. All stages of host eggs can be successfully parasitized, and we show here that either the parasitoid larva or the wasp assures that the larva eventually is located in the host's haemocoel. (1) When freshly laid eggs, up to almost 1-day-old, are parasitized, the parasitoid hatches while still in the yolk and enters the host either after waiting or immediately through the dorsal opening. (2) When 1-2-day old eggs are parasitized, the host embryo has accomplished final dorsal closure and is covered by an embryonic cuticle when the parasitoid hatches; in this case the parasitoid larva bores with its moving abdominal tip into the host. (3) When 2.5-3.5-day-old eggs are parasitized, the wasp oviposits directly into the haemocoel of the host embryo; from day 2 to 2.5 the embryo is still very small and the wasps, after probing, often restrain from oviposition for a few hours. PMID- 15749112 TI - Triggering and persistence of trail-laying in foragers of the ant Lasius niger. AB - In the ant Lasius niger, the ability to ingest their own desired volume is the key criterion that rules the recruiting behaviour of scouts. This volume acts as a threshold triggering the trail-laying response of foragers. In this paper, we show that this desired volume is specific to each individual and is kept constant over successive trips to a food source. This individual specificity contrasts with the variability of all individual desired volumes within the colony. In this study, it is also shown that, among L. niger foragers, 14% never participate in the formation of the chemical pathway and never lay a trail over successive trips. Among the others foragers, interindividual differences in the persistence of trail-laying behaviour over successive trips are observed but do not rely on an individual specialisation, in which some ants would lay a trail more frequently and persistently than other scouts. We discuss how an individual in the foraging behaviour can play an essential role in the regulation of food retrieval dynamics. PMID- 15749113 TI - PBAN selective antagonists: inhibition of PBAN induced cuticular melanization and sex pheromone biosynthesis in moths. AB - A D-Phe scan (sequential D-Phe replacement) library of linear peptides, synthesized on the basis of a slightly modified active sequence of PBAN (YFSPRL amide) was employed to detect potential inhibitors of cuticular melanization in Spodoptera littoralis larvae and to compare their stimulatory and inhibitory melanization activity with their pheromonotropic agonistic and antagonistic activities. A quantitative melanotropic assay was used to monitor the extent of cuticular melanization elicited by Hez-PBAN1-33NH2 in S. littoralis larvae in the presence and absence of the D-Phe peptides. The data revealed the presence of two partial melanotropic antagonists, and disclosed the presence of selective pure melanotropic agonists and pure pheromonotropic antagonists indicating differences in the inhibitory and stimulatory patterns of the library with respect to both activities. The differences between the pheromonotropic and melanotropic inhibitory patterns of the peptides hints at the possibility that sex pheromone biosynthesis in the pheromone gland of Heliothis peltigera females and induction of cuticular melanization in S. littoralis may be mediated by different receptors (that may result either from presence of different receptor sub-types or may reflect species differences in receptor structure and/or properties) despite the fact that they are induced by the same peptide (PBAN1-33NH2). PMID- 15749114 TI - Social versus individual behaviour: a comparative approach to thermal behaviour of the honeybee (Apis mellifera L.) and the American cockroach (Periplaneta americana L.). AB - To study the relationship between the individual and social thermoregulatory behaviour, we used honeybee workers and American cockroaches. Single insects or groups of 10-20 individuals were placed in a temperature gradient chamber, and their thermal preference was recorded for 48 h under natural summer photoperiod. Single bees showed diurnal changes in selected ambient temperature, which culminated at 14:00 reaching 34+/-2 degrees C, and then slowly decreased, reaching a nocturnal minimum of 28+/-2 degrees C at 04:00. In contrast, the zenith of temperature selected by groups of bees (31+/-1 degrees C) was reached at 04:00 and the nadir (29+/-2 degrees C) was recorded at 14:00. Groups of bees clustered together during the night time, and dispersed during intense day time activity. Such changes were absent in groups of cockroaches. Cockroaches selected an ambient temperature of 30+/-1 degrees C both during day and night. In conclusion, there is a striking analogy in the diurnal thermal behaviour between a colony of bees and mammals. During their nychthemeral rest phase, both of them select higher temperatures than during the activity phase and, simultaneously, they reduce their overall surface area of heat loss to conserve metabolic heat. Therefore, the colony behaves as a homeothermic superorganism. In contrast, a single bee, isolated from the colony, utilizes a heterothermic strategy to save energy for a morning warm up. PMID- 15749115 TI - Quantification of vitellogenin-mRNA during maturation and breeding of a burying beetle. AB - Burying beetles (Nicrophorus orbicollis) are unusual in that to breed they require an unpredictable and valuable resource, a small carcass. Thus the timing of reproduction is unpredictable and beetles' physiological response must be fast. We hypothesized that their pattern of vitellogenin (Vg) synthesis might reflect these requirements. We examined the expression of two Vg genes (sequenced for this study) during sexual maturation and through a reproductive bout. Vg mRNA, juvenile hormone (JH) titers, ovarian development, and hemolymph concentrations of Vg were quantified in the same individuals. All four variables gradually increased during maturation to peak 15-20 days after eclosion. Twelve hours after the discovery of a carcass, a few hours before oviposition, mRNA was high, hemolymph Vg had decreased, JH and ovarian weight had increased. After oviposition, mRNA was low, hemolymph Vg concentrations and JH were high. This is consistent with our hypothesis that beetles produce and store Vg in the hemolymph prior to the discovery of a breeding resource and replace it quickly. Partial regression of these variables (with the effect of time removed) indicated that JH was not correlated with mRNA, hemolymph Vg, or ovarian weight at any time. Thus the role of JH as a gonadotropin remains unclear. PMID- 15749116 TI - The adipokinetic hormones of Odonata: a phylogenetic approach. AB - Adipokinetic neuropeptides from the corpora cardiaca of the major families of all three suborders of the Odonata were identified by one or more of the following methods: (1) Isolation of the peptides from a methanolic extract of the corpora cardiaca by liquid chromatography, peak monitoring by fluorescence of the Trp residue and comparison of the retention time with those of known synthetic peptides of Odonata. (2) Hyperlipaemic bioassays of the HPLC-generated fractions either in Locusta migratoria or, in a few cases, in Anax imperator or Orthetrum julia. (3) Sequencing of the isolated, bioactive HPLAC fraction by Edman degradation. (4) Mass spectrometric measurement of the isolated, bioactive fraction. Sequence assignment revealed that the investigated Odonata species always contain only one adipokinetic peptide. This is always an octapeptide. The suborder Zygoptera contains the peptide code-named Psein-AKH, the Anisozygoptera and the families Aeshnidae, Cordulegastridae and Macromiidae of the Anisoptera contain Anaim-AKH, whereas Gomphidae, Corduliidae (with the exception of Syncordulia gracilis) and Libellulidae contain Libau-AKH; one species of Libellulidae has Erysi-AKH, a very conservative modification of Libau-AKH (one point mutation). When these structural data are interpreted in conjunction with existing phylogenies of Odonata, they support the following: (1) Zygoptera are monophyletic and not paraphyletic. (2) Anisozygoptera and Anisoptera are sister groups and contain the ancestral Anaim-AKH which is independently and convergently mutated to Libau-AKH in Gomphidae and Libellulidae. (3) The Corduliidae are of special interest. Only Corduliidae sensu stricto appear to contain Libau-AKH, other species placed into this family by most authorities contain the ancestral Anaim-AKH. Possibly, assignments of AKHs can untangle the paraphyly of this family. PMID- 15749117 TI - CD13/aminopeptidase N and murine cytomegalovirus infection. AB - CD13/aminopeptidase N is a membrane-bound metalloproteinase implicated in human cytomegalovirus (HCMV) infection and pathogenesis. Anti-CD13 antibodies can neutralize HCMV infectivity, and HCMV viremia after bone marrow transplantation induces anti-CD13 autoantibodies which correlate with development of chronic graft vs. host disease. We examined whether murine CD13/APN was similarly implicated in murine cytomegalovirus (MCMV) disease. MCMV infection did induce anti-CD13 antibodies in mice in a strain-specific manner. ICR and 129S mice developed high titers of anti-CD13 antibodies and anti-MCMV antibodies after MCMV infection, whereas CBA and CBAxC57BL/6 f1 hybrid mice produced antibodies against MCMV only. Unlike HCMV, no evidence was found for a correlation between host cell CD13/APN expression and infection, or for the presence of CD13/APN on MCMV particles, although APN inhibitors decreased MCMV plaque formation. Reproduction of CD13/APN autoantibody production in the murine system should make it possible to determine if these antibodies contribute to CMV pathogenesis. PMID- 15749118 TI - HCV associated glomerulopathy in Egyptian patients: clinicopathological analysis. AB - BACKGROUND: Hepatitis C virus (HCV) infection in Egypt has reached an epidemic proportion and is associated with many extra hepatic manifestations; Glomerulonephritis (GN) is one of the most consequences of HCV infection often resulting in end stage renal disease in some cases. Detection of viral genome or particles within the kidney biopsies from HCV-infected patients has proven to be difficult. Histological characterization of renal lesions still represents a major challenge. The aim of our work was to describe the histological pattern of HCV-associated nephropathy. METHODS: Fifty Patients--out of 233--presented to Mansoura Urology and Nephrology clinic with manifestations of glomerular disease were screened for HCV antibodies by a 3rd generation ELISA test. Those tested positive for HCV antibodies were confirmed by PCR for HCV-RNA and subjected to more detailed clinical, biochemical and histological study. Kidney biopsies and in appropriate cases liver biopsies were examined by LM and electron microscopy (EM). RESULTS: Histological study of renal biopsies revealed membranoproliferative (MPGN) type 1 to be the most common lesion encountered (54%), followed by focal segmental glomerulosclerosis (FSGS) (24%), mesangioproliferative GN (18%), membranous nephropathy (MN) (4%) in that order. EM examinations of renal biopsies were successful in identifying HCV like particles in frozen renal tissue. CONCLUSION: HCV-associated glomerulopathy is a distinct category of glomerulonephritis. Results of LM showed some peculiar features. In addition, we were successful in location and detection of HCV particles in renal tissues by EM. PMID- 15749119 TI - Maturation of a tetravirus capsid alters the dynamic properties and creates a metastable complex. AB - The assembly of monomeric protein subunits into a viral capsid is a finely tuned molecular process. In response to subtle changes in environmental conditions, this supramolecular complex can dramatically reorganize. Defining the forces that control this structure and the cooperative action of subunits has implications for biology and nanotechnology. The small icosahedral RNA tetravirus family members Nudaurelia omega capensis (NomegaV) and Helicoverpa armigera stunt virus (HaSV) can be purified as provirions, and maturation to capsids can be induced by a drop in pH. In this study, a comparison of capsid secondary structure using FT IR revealed that the procapsid has more alpha-helical content than the capsid, supporting the proposal that helix to coil transition may be important for maturation. The dynamic properties of the two states were probed using limited proteolysis and peptide mass mapping to identify regions of significant flexibility. Interestingly, the initial sites of protease cleavage were the N and C terminal domains that are internal in high-resolution models, and to inter subunit surfaces. Further comparison of the two particle forms using FT-IR revealed that in response to thermal stress, the provirion disassembles and unfolds in a cooperative manner over a narrow temperature range (approximately 5 degrees C). Paradoxically, the capsid form, which is stable in a wide range of pH and ionic conditions and is more resistant to proteolysis, responds to thermal stress at a lower temperature than the procapsid form. This suggests that a metastable state is the end product of assembly. PMID- 15749120 TI - An infectious West Nile virus that expresses a GFP reporter gene. AB - West Nile virus is a mosquito-borne, neurotropic flavivirus that causes encephalitis in humans and animals. Since being introduced into the Western hemisphere in 1999, WNV has spread rapidly across North America, identifying this virus as an important emerging pathogen. In this study, we developed a DNA launched infectious molecular clone of WNV that encodes a GFP reporter gene. Transfection of cells with the plasmid encoding this recombinant virus (pWNII GFP) resulted in the production of infectious WNV capable of expressing GFP at high levels shortly after infection of a variety of cell types, including primary neurons and dendritic cells. Infection of cells with WNII-GFP virus was productive, and could be inhibited with both monoclonal antibodies and interferon beta, highlighting the potential of this system in the development and characterization of novel inhibitors and therapeutics for WNV infection. As expected, insertion of the reporter gene into the viral genome was associated with a reduced rate of viral replication, providing the selective pressure for the development of variants that no longer encoded the full-length reporter gene cassette. We anticipate this DNA-based, infectious WNV reporter virus will allow novel approaches for the study of WNV infection and its inhibition both in vitro and in vivo. PMID- 15749121 TI - Biosynthesis of the IFN-gamma binding protein of ectromelia virus, the causative agent of mousepox. AB - Ectromelia virus (ECTV), the causative agent of mousepox, expresses an extracellular interferon-gamma binding protein (IFN-gammaBP) with homology to the ligand binding domains of the IFN-gamma high affinity receptor (IFN-gammaR1). Unlike the cellular receptor, the IFN-gammaBP binds IFN-gamma from several species. The IFN-gammaBP is synthesized early after infection, accumulating in the extracellular milieu as dimers composed of two protein species with Mr of 34.6 or 33.0 kDa. Homodimers are covalently linked by an interchain disulphide bond at position 216. The IFN-gammaBP has complex N-linked oligosaccharides at positions 41 and 149 as determined by site-directed mutagenesis and glycosidase treatment. Glycosylation at position 41 is required for secretion from mammalian cells and may play a role in the activity of the IFN-gammaBP. Glycosylation at position 149 is not required for secretion, and the lack of glycosylation at this site does not diminish ligand binding as measured by surface plasmon resonance (SPR) and ELISA. PMID- 15749122 TI - The vomeronasal chemosensory system as a route of neuroinvasion by herpes simplex virus. AB - We have investigated the potential of neurotropic microbes to invade the central nervous system (CNS) via the peripheral nervous system. Herpes simplex virus type 1 (HSV-1) strain KH6 and herpes simplex virus type 2 (HSV-2) strain 186 were found to infect chemosensory neurons in the vomeronasal organ (the pheromone detector) following intranasal inoculation of mice. HSV-1 strain KH6 infection was further transmitted to the accessory olfactory bulb (first relay), the medial amygdala (second relay), and the bed nucleus of the stria terminalis and the ventromedial hypothalamus (third relay). HSV-1 strain KH6 also targeted the olfactory and trigeminal systems. HSV-2 strain 186 predominantly attacked the brainstem including the trigeminal system. While both viruses did not induce apoptosis in infected chemosensory neurons, they did in infected brain tissue. These results suggest that neurotropic viruses can invade the brain by infecting vomeronasal chemosensory neurons and that the restrained induction of apoptosis in the infected neurons may facilitate viral transmission to the CNS. PMID- 15749123 TI - Ethanol potentiates HIV-1 gp120-induced apoptosis in human neurons via both the death receptor and NMDA receptor pathways. AB - Neuronal loss is a hallmark of AIDS dementia syndromes. Human immunodeficiency virus type I (HIV-1)-specific proteins may induce neuronal apoptosis, but the signal transduction of HIV-1 gp120-induced, direct neuronal apoptosis remains unclear. Ethanol (EtOH) is considered to be an environmental co-factor in AIDS development. However, whether EtOH abuse in patients with AIDS increases neuronal dysfunction is still uncertain. Using pure, differentiated, and post-mitotic NT2.N-derived human neurons, we investigated the mechanisms of HIV-1 and/or EtOH related direct neuronal injury and the molecular interactions between HIV-1 specific proteins and EtOH. It was demonstrated that NT2.N neurons were susceptible to HIV-1 Bal (R5-tropic strain) gp120-induced direct cell death. Of importance, EtOH induced cell death in human neurons in a clinically-relevant dose range and EtOH strongly potentiated HIV-1 gp120-induced neuronal injury at low and moderate concentrations. Furthermore, this potentiation of neurotoxicity could be blocked by N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B) antagonists. We analyzed human genomic profiles in these human neurons, using Affymetrix genomics technology, to elucidate the apoptotic pathways involved in HIV-1- and EtOH-related neurodegeneration. Our findings indicated significant over-expression of selected apoptosis functional genes. Significant up-regulation of TRAF5 gene expression may play an essential role in triggering potentiation by EtOH of HIV-1 gp120-induced neuronal apoptosis at early stages of interaction. These studies suggested that two primary apoptotic pathways, death receptor (extrinsic) and NMDA receptor (intrinsic)-related programmed cell-death pathways, are both involved in the potentiation by EtOH of HIV-1 gp120-induced direct human neuronal death. Thus, these data suggest rationally-designed, molecular targets for potential anti-HIV-1 neuroprotection. PMID- 15749124 TI - Identification of a critical neutralization determinant of severe acute respiratory syndrome (SARS)-associated coronavirus: importance for designing SARS vaccines. AB - The spike (S) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is not only responsible for receptor binding, but also a major antigenic determinant capable of inducing protective immunity. In this study, we demonstrated that the receptor-binding domain (RBD) of S protein is an important immunogenic site in patients with SARS and rabbits immunized with inactivated SARS-CoV. Serum samples from convalescent SARS patients and immunized rabbits had potent neutralizing activities against infection by pseudovirus expressing SARS CoV S protein. Depletion of RBD-specific antibodies from patient or rabbit immune sera by immunoadsorption significantly reduced serum-mediated neutralizing activity, while affinity-purified anti-RBD antibodies had relatively higher potency neutralizing infectivity of SARS pseudovirus, indicating that the RBD of S protein is a critical neutralization determinant of SARS-CoV during viral infection and immunization. Two monoclonal antibodies (1A5 and 2C5) targeting at the RBD of S protein were isolated from mice immunized with inactivated SARS-CoV. Both 1A5 and 2C5 possessed potent neutralizing activities, although they directed against distinct conformation-dependant epitopes as shown by ELISA and binding competition assay. We further demonstrated that 2C5, but not 1A5, was able to block binding of the RBD to angiotensin-converting enzyme 2 (ACE2), the functional receptor on targeted cells. These data provide important information for understanding the antigenicity and immunogenicity of SARS-CoV and for designing SARS vaccines. PMID- 15749125 TI - cis-Acting sequences required for coat protein binding and in vitro assembly of Potato virus X. AB - The 5' region of Potato virus X (PVX) RNA containing an AC-rich single-stranded region and stem-loop 1 (SL1) has been shown to be important for PVX replication (Miller, E.D., Plante, C.A., Kim, K.-H., Brown, J.W., Hemenway, C., 1998. Stem loop structure in the 5' region of potato virus X genome required for plus-strand RNA accumulation. J. Mol. Biol. 284, 591-608.). Here, we describe the involvement of SL1 for binding to the PVX coat protein (CP) using an in vitro assembly system and various deletion mutants of the 5' region of PVX RNA. Internal and 5' terminal deletions of the 5'-nontranslated region of PVX RNA were assessed for their effects on formation of assembled virus-like particles (VLPs). Mutant RNAs that contain the top region of SL1 or sequences therein bound to CP to form VLPs. In contrast, transcripts of mutants that disrupt SL1 RNA structure were unable to form VLPs. SELEX was used to further confirm the specific RNA recognition of PVX CP using RNA transcripts containing randomized sequences of the upper portion of SL1. Wild-type (wt) sequences along with many other sequences that resemble SL1 structure were selected after fourth and fifth rounds of SELEX (27.0% and 44.4%, respectively). RNA transcripts from several SELEX winners that are predicted to form stable stem-loop structures very closely resembling wt PVX SL1 VLPs. RNA transcripts not predicted to form secondary structures similar to SL1 did not form VLPs in vitro. Taken together, our results suggest that RNA secondary structural elements within SL1 and/or sequences therein are crucial for formation of VLPs and are required for the specific recognition by the CP subunit. PMID- 15749127 TI - Programmed remodeling of hyperacetylated histone H4 and H3 organization on the SV40 genome during lytic infection. AB - The presence of nucleosomes containing hyperacetylated histone H4 and H3 on the early, late, and promoter regions of the SV40 genome in chromosomes isolated 30 min, 8 h, and 48 h post-infection was determined by chromatin immunoprecipitation (ChIP) analysis with PCR amplification of fragmented SV40 chromatin using two complementary strategies. In chromosomes isolated at 30 min post-infection hyperacetylated H4 was found intermittently in all the three regions with no preference for one region over the other. In contrast, hyperacetylated H3 was organized primarily within the promoter region and occasionally elsewhere. At 8 h post-infection, nucleosomes with both hyperacetylated H4 and H3 were found regularly associated with all three regions of the SV40 genome. Finally, in SV40, chromosomes isolated 48 h post-infection hyperacetylated H4 and H3 were found frequently associated with all regions of the chromosome although hyperacetylated H4 was preferentially associated with the late region. The changing patterns of organization of hyperacetylated histones in SV40 chromosomes during the course of a lytic infection presumably reflects the different biological functions of the SV40 chromatin at each of the time points. PMID- 15749126 TI - Substitutions of conserved amino acids in the receptor-binding domain of the spike glycoprotein affect utilization of murine CEACAM1a by the murine coronavirus MHV-A59. AB - The host range of the murine coronavirus (MHV) is limited to susceptible mice and murine cell lines by interactions of the spike glycoprotein (S) with its receptor, mCEACAM1a. We identified five residues in S (S33, L79, T82, Y162 and K183) that are conserved in the receptor-binding domain of MHV strains, but not in related coronaviruses. We used targeted RNA recombination to generate isogenic viruses that differ from MHV-A59 by amino acid substitutions in S. Viruses with S33R and K183R substitutions had wild type growth, while L79A/T82A viruses formed small plaques. Viruses with S33G, L79M/T82M or K183G substitutions could only be recovered from cells that over-expressed a mutant mCEACAM1a. Viruses with Y162H or Y162Q substitutions were never recovered, while Y162A viruses formed minute plaques. However, viruses with Y162F substitutions had wild type growth, suggesting that Y162 may comprise part of a hydrophobic domain that contacts the MHV-binding site of mCEACAM1a. PMID- 15749128 TI - Studies on the cross-clade and cross-species conservation of HIV-1 Gag-specific CD8 and CD4 T cell responses elicited by a clade B DNA/MVA vaccine in macaques. AB - Here, we evaluate the T cell responses raised by our HIV-1 clade B DNA/MVA vaccine for recognition of a HIV-1 circulating recombinant form (CRF) AG Gag sequence (CRF-02). The cross-clade activity for the AG sequence was better conserved for CD8 than CD4 T cells. CD8 T cells exhibited 75% conservation for height and 83% conservation for breadth, whereas CD4 responses exhibited 45% conservation for height and 50% conservation for breadth. Five CD8 epitopes and 8 CD4 epitopes were mapped. Three of the 5 CD8 epitopes and 2 of the 8 CD4 epitopes were conserved across multiple HIV-1 clades. Impressively, all of the CD8 epitopes and half of the CD4 epitopes have been reported for human infections. Our results demonstrate that the clade B DNA/MVA HIV vaccine elicits T cell responses against epitopes that are conserved in multiple clades and recognized by humans and macaques. PMID- 15749130 TI - Increasing colorectal cancer screening among individuals in the carpentry trade: test of risk communication interventions. AB - BACKGROUND: Individuals in the carpentry trade, due to lifestyle habits and occupational exposures, may be at above-average risk for colorectal cancer (CRC). Based on the literature which suggests that increasing perceived risk motivates behavior change, we report on the effectiveness of four risk-communication interventions targeted to increase initial, yearly and repeat fecal occult screening (FOBT) among carpenters (N = 860) over a 3-year period. METHODS: Our 2 x 2 factorial design intervention study varied two dimensions of providing CRC risk factor information: (1) type of risk factor-one set of interventions emphasized three basic risk factors (age, family history and polyps); the other set emphasized a comprehensive set of risk factors including basic, lifestyle, and occupational factors, and (2) tailoring/not tailoring risk factor information. Participants were provided FOBTs. Outcomes were the proportion of returned FOBTs. RESULTS: Varying the amount and intensity of delivering CRC risk factors information affected neither risk perceptions nor initial, yearly, or repeat screening. However, yearly and repeat screening rates were greater among participants who received interventions addressing comprehensive set of risk factors, especially with increasing age. CONCLUSIONS: Tailoring on several CRC risk factors appears insufficient to increase and sustain elevated perceptions of CRC risks to promote screening. PMID- 15749129 TI - Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). AB - Inactivated severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been tested as a candidate vaccine against the re-emergence of SARS. In order to understand the efficacy and safety of this approach, it is important to know the antibody specificities generated with inactivated SARS-CoV. In the current study, a panel of twelve monoclonal antibodies (mAbs) was established by immunizing Balb/c mice with the inactivated BJ01 strain of SARS-CoV isolated from the lung tissue of a SARS-infected Chinese patient. These mAbs could recognize SARS-CoV-infected cells by immunofluorescence analysis (IFA). Seven of them were mapped to the specific segments of recombinant spike (S) protein: six on S1 subunit (aa 12-798) and one on S2 subunit (aa 797-1192). High neutralizing titers against SARS-CoV were detected with two mAbs (1A5 and 2C5) targeting at a subdomain of S protein (aa 310-535), consistent with the previous report that this segment of S protein contains the major neutralizing domain. Some of these S specific mAbs were able to recognize cleaved products of S protein in SARS-CoV infected Vero E6 cells. None of the remaining five mAbs could recognize either of the recombinant S, N, M, or E antigens by ELISA. This study demonstrated that the inactivated SARS-CoV was able to preserve the immunogenicity of S protein including its major neutralizing domain. The relative ease with which these mAbs were generated against SARS-CoV virions further supports that subunit vaccination with S constructs may also be able to protect animals and perhaps humans. It is somewhat unexpected that no N-specific mAbs were identified albeit anti-N IgG was easily identified in SARS-CoV-infected patients. The availability of this panel of mAbs also provided potentially useful agents with applications in therapy, diagnosis, and basic research of SARS-CoV. PMID- 15749131 TI - Focused interviews with Sudanese Americans: perceptions about diet, nutrition, and cancer. AB - BACKGROUND: This study explored perceptions regarding diet, nutrition and cancer among Sudanese-Americans; and generated information for response options, components to questions, and actual questions for a pre-conceptualized diet, nutrition and cancer-related questionnaire to be used on this population. METHODS: In a descriptive, qualitative study, data were collected from four focus group discussions with Sudanese-Americans (N = 18). RESULTS: Most participants were fearful of cancer, associating it with death. Stress, genetics, and pesticides in vegetables were given as major causes of cancer. Fatalism towards cancer pervaded among a few participants. Male participants did not believe that dietary factors could be helpful in cancer prevention, while the females believed fried, fatty, smoked and fast foods could lead to cancer. Factors perceived as influencing food choices included money, cultural traditions and social class. Cultural tradition was perceived as the major barrier to changing food habits. Frying, stewing, grilling, boiling and fermenting were the most common food preparation methods reported. CONCLUSION: Perceptions regarding dietary practices, nutrition and cancer among Sudanese-Americans, some of which might have been overlooked were identified. The focus groups generated response options, components to questions, and actual questions for a pre-conceptualized diet, nutrition and cancer-related questionnaire to be used on Sudanese Americans. PMID- 15749132 TI - Calculation of population attributable risk for bidi smoking and oral cancer in south Asia. AB - BACKGROUND: Bidi smoking, which is widely prevalent in India and in other south Asian countries, increases the risk of oral cancer as observed in case-control studies and metaanalysis. However, population attributable risk percent (PAR%) has not been determined yet. MATERIALS AND METHODS: Twelve case-control studies conducted in India, Pakistan, and Sri Lanka, which included information on bidi smoking and oral cancer, were analyzed countrywise to estimate PAR%. RESULTS: The cumulative cases and controls were 4778 and 6271, respectively, based on 10 case control studies conducted in India. Among the cases, 49.1% were bidi smokers and 7.7% cigarette smokers, while they were 19.9% and 10.3%, respectively, among controls. Pooled odds ratio (OR) of bidi smoking for oral cancer was 3.3 [95% confidence interval (CI), 3.0-3.6] and 2.6 (95% CI 1.8-3.8), respectively, based on fixed- and random-effects model. Cigarette smoking, on the other hand, did not show any significant association. PAR% of bidi smoking for oral cancer ranged from 4.7% to 51.6% on individual study basis, while they were 31.4% and 24.1%, respectively, based on OR derived from fixed- and random-effects models. PAR% was 5.8% and 8.7% based on single study estimate from Pakistan and Sri Lanka, respectively. CONCLUSIONS: Bidi smoking is considered to account for a sizeable number of oral cancers in south Asian countries, which implies that cessation programs should be formulated and implemented vigorously. PMID- 15749133 TI - Cancer screening among Latino subgroups in the United States. AB - BACKGROUND: Relying upon the Health Belief Model and a behavioral model of health care utilization, the purpose of this study was to examine current adherence to cancer screening among Latino subgroups. METHODS: Using data from the 2000 National Health Interview Survey, 5377 Latinos were surveyed for their use of Pap smear, mammogram, breast self-examination and the clinical breast exam among women, prostate specific antigen test among men, and the fecal occult blood test, sigmoidoscopy, colonoscopy, and proctoscopy among both men and women. Using sampling weights, multivariate logistic regression models were used to assess screening use. RESULTS: Dominican women had 2.4 times greater likelihood of having had mammography than other Latinos. In addition, Latinas aged 50-69, who had more years of education, a personal history of cancer, who were not current smokers, had health insurance, had visited a primary care provider over the past 12 months, and had at least one other screening test had greater use of mammography. Younger age, marriage, greater acculturation, visits to a primary care provider, health insurance, and the use of other cancer screening tests predicted the uptake of the Pap smear. Latinas were more likely to use a CBE if they were younger, had a Bachelor's degree, a personal history of cancer, were more acculturated, had visits to a primary care provider over the past 12 months, and used other cancer screening tests. Puerto Ricans, Central or South Americans had half the likelihood of having colorectal cancer screening than other groups. Ages between 50 and 69, male sex, marriage, history of visiting a health care provider, and use of other screening tests predicted use of the FOBT. Older age, greater education, male sex, history of visiting a health care provider in the previous year, use of other screening tests, and better health status influenced the uptake of endoscopy for colorectal cancer screening. Cuban males had fivefold greater utilization of PSA testing. Additionally, PSA use among Latinos was predicted by older age, history of visiting a primary care provider in the past 12 months, and use of other screening tests. CONCLUSIONS: Cancer screening programs must take into account differences among Latinos in age, gender, educational levels, marital status, cancer history, risk behaviors, insurance, health status and health services utilization. PMID- 15749135 TI - Cancer incidence among Mormons and non-Mormons in Utah (United States) 1995-1999. AB - BACKGROUND: Population-based Utah Cancer Registry data were linked with Latter day Saint (LDS or Mormon) Church membership records to obtain site-specific cancer incidence for LDS and non-LDS populations in Utah during 1995-1999. METHODS: Analyses were based on 27,631 incident cases of cancer identified among whites. Restriction to whites was made because of the small number of nonwhites, approximately 5%, in the state during the study period. The direct method was used to age-adjust the rates to the 2000 U.S. standard population. RESULTS: Significantly lower cancer incidence rates per 100,000 were observed among LDS compared with non-LDS males (287.2 vs. 321.1) and females (247.7 vs. 341.0). The lower rates are primarily explained by smoking-related cancers and female breast cancer. If the overall cancer incidence rate in LDS had occurred in the non-LDS population, 2.9% or 421 fewer cases would have occurred among males and 7.9% or 1,025 fewer cases would have occurred among females during the study period. CONCLUSIONS: Given our current knowledge of risk factors for cancer, differences between LDS and non-LDS in smoking for males and smoking and sexual and reproductive behaviors in females primarily explain the lower risk of cancer in LDS populations. PMID- 15749134 TI - Prevalence and predictors of multiple behavioral risk factors for colon cancer. AB - OBJECTIVES: This paper examines the prevalence of behavioral risk factors for colorectal cancer (CRC) (e.g., red meat consumption, fruit and vegetable intake, multivitamin intake, alcohol, smoking, and physical inactivity), co-occurrence among these behaviors, and motivation for change among patients at increased risk. METHODS: The study sample included 1,247 patients with recent diagnosis of adenomatous colorectal polyps. Within 4 weeks following the polypectomy, participants completed a baseline survey by telephone. RESULTS: Sixty-six percent of participants had not been diagnosed with polyps before. Fifty-eight percent of the sample had red meat as a risk factor, 63% had fruit and vegetable consumption as a risk factor, 54% did not take a daily multivitamin, and 44% had physical activity as a risk factor. In contrast, only 9% of the sample had alcohol consumption as a risk factor and only 14% were current smokers. The prevalence of the six individual risk factors was combined into an overall multiple risk factor score (MRF). The average number of risk factors was 2.43. Men, those with a high school education or below, those reporting fair or poor health status, and those with less self-efficacy about risk factor change had more risk factors. CONCLUSIONS: There is a need for multiple risk factor interventions that capitalize on natural intersections among intra- and interpersonal factors that maintain them. PMID- 15749136 TI - Provision of health counseling in office-based practices and hospital outpatient clinics. AB - OBJECTIVE: To compare the rates of health counseling provided during primary care visits in two different types of ambulatory care settings. METHODS: Secondary analysis of the 2000 National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS). RESULTS: Of the estimated 722 million adult ambulatory care visits during 2000, 90.8% were made to office-based physician practice settings and 9.2% to hospital-based outpatient departments. Consistent with previous reports, the demographic profile of patients who seek primary care in hospital outpatient departments differs from those seen in office based practices. Provision of health counseling for exercise [OR = 1.4; 95% confidence intervals (CI): 1-1.8], diet (OR = 1.6; 95% CI: 1.2-2.3), breast self exam (OR = 2; 95% CI: 1.1-3.6) and stress management (OR = 1.7; 95% CI: 1-2.7) during patient visits was more likely to be reported in the office-based practices than in hospital outpatient clinics. The visit-based rates of health counseling for HIV/STD prevention, tobacco use, mental health or injury prevention were low in both settings. CONCLUSIONS: There is opportunity to improve rates of preventive counseling in primary care settings and to reduce disparities that exist. Identifying the reasons for these disparities and effective interventions will be important steps in providing equitable care in the area of preventive health counseling. PMID- 15749137 TI - Can current physical activity act as a reasonable proxy measure of future physical activity? Evaluating cross-sectional and passive prospective designs with the use of social cognition models. AB - BACKGROUND: The standard methodological approach for evaluating social cognitive theories when predicting physical activity behavior is the passive prospective/longitudinal survey design. Although this design is logical, a cross sectional design may be a cost-effective alternative if the relationships between social cognitive constructs and physical activity are relatively stable. The purpose of this study was to evaluate the utility of a concurrent measure of physical activity used in a cross-sectional design in comparison to the standard prospective measure. METHODS: This study included two 6-month prediction time periods, between 1997 and 1998, for the purpose of analysis replication, and the theory of planned behavior, the transtheoretical model, protection motivation theory, and social cognitive theory as the models of interest in a population sample (N = 703). RESULTS: Results showed trivial (69% of tests; q < 0.10) to small (31% of tests; q = 0.11-0.18) differences in the correlations between social cognitive constructs and vigorous physical activity occur when using a cross-sectional or prospective design. The cross-sectional design estimated slightly larger coefficients than the prospective design. CONCLUSIONS: It appears that a measure of concurrent physical activity included in a cross-sectional design can act as a reasonable proxy measure of future behavior measured in a passive prospective/longitudinal design. These findings support the use of cross sectional designs when researchers seek a standard correlational investigation of physical activity and social cognitive constructs with the possibility that coefficients may be slightly biased upwards. PMID- 15749138 TI - Physicians' attitudes and preventive care delivery: insights from the DOPC study. AB - BACKGROUND: Interventions that modify physician attitudes to enhance preventive service delivery are common, yet other factors may be relatively more important in determining whether these services are provided. We assessed associations between physicians' attitudes and delivery of preventive care, compared with factors related to the patient, visit, or practice. METHODS: One hundred twenty eight primary care physicians rated the importance of five preventive services and their effectiveness at delivering them. We assessed whether their patients had received cervical smears, prostate-specific antigen (PSA) testing, smoking cessation advice, recommendation to use aspirin to prevent myocardial infarction, or weight-maintenance counseling, when appropriate. Multilevel models assessed associations between physician attitudinal characteristics and a patient's likelihood of being up to date for each service. RESULTS: Importance of PSA screening and tobacco cessation counseling were weakly associated with patients' receipt of preventive care; no association between attitudes and other services was observed. Factors such as having a visit for well care and use of prevention flowcharts were associated with delivery of preventive services to a greater extent. CONCLUSIONS: Physicians' attitudes toward prevention are necessary, but not sufficient in ensuring the delivery of preventive services. Future interventions should address visit- and practice-specific factors more closely associated with preventive care. PMID- 15749139 TI - Characteristics of glycemic control in elite power and endurance athletes. AB - A previous study has shown that former elite power athletes exhibited significantly greater relative risk in diabetes than that of former elite endurance athletes. It is unknown whether insulin sensitivity in elite young healthy power athletes is lower than that in elite young endurance athletes. This study includes two parts, part I and part II. In the part I of this study, an oral glucose tolerance test was performed in all of the elite juvenile track athlete subjects, specializing either in short-distance racing (jSD, N = 13, aged 12.5 +/- 0.37 years) or in long-distance racing (jLD, N = 13, aged 12.6 +/- 0.42 years). In the part II of this study, we recruited elite adult swimmers and divided them into two groups according to their specialty in swimming race distance: long-distance (aLD, N = 10, age 20.3 +/- 1.32) and short-distance groups (aSD, N = 10, age 20.2 +/- 1.31). Insulin sensitivity was significantly lower in the jSD group than that in the jLD group, as indicated by the area under the curves of insulin and glucose following a 75-g oral glucose load. Fasting plasma LDL-C and total cholesterol levels in the jSD group were significantly greater than those in the jLD group. The result of the part II of this study, similar to the result of the part I, shows that insulin sensitivity in aSD swimmers was significantly lower than that in aLD swimmers. LDL-C, total cholesterol, and triglyceride levels were also found higher in aSD swimmers than in those of aLD swimmers. These new findings implicate that the genetic makeup associated with exceptional power or endurance performance of elite athletes could also reflect on their metabolic characteristics; elite power athletes appear to be more insulin resistant than elite endurance athletes. PMID- 15749140 TI - Self-reported diabetes and posttraumatic stress disorder among adults in the community. AB - OBJECTIVE: To determine the association between self-reported diabetes and posttraumatic stress disorder (PTSD) among adults in the community and to investigate the specificity of this link. METHOD: Data were drawn from the National Comorbidity Survey (NCS), a household probability sample of adults ages 15-54 in the United States. Multiple logistic regression analyses were used to investigate the association between self-reported diabetes and PTSD (past 12 month prevalence), adjusted for differences in sociodemographic characteristics. RESULTS: Self-reported diabetes was associated with an increased likelihood of PTSD [OR = 2.3 (1.02, 5.21)], which persisted after adjusting for differences in sociodemographic characteristics. Self-reported diabetes was not associated with a significantly increased likelihood of any other mental disorders. CONCLUSIONS: These findings are consistent with and extend previous data suggesting that there is an association between self-reported diabetes and PTSD by showing that this link appears to have some specificity and is generalizable to adults in the community. These data do not address the possible mechanisms of this association. Our results do not support a link between depression and diabetes, which has been previously reported. Replication of these results is needed with longitudinal, epidemiologic data, which include ages of onset and physiologic data in diagnosis of diabetes. If these findings are replicated, further investigation into the possible mechanisms of this association may be fruitful. PMID- 15749141 TI - Is advice to stop smoking from a midwife stressful for pregnant women who smoke? Data from a randomized controlled trial. AB - BACKGROUND: There are no randomized trials examining whether intensive advice to pregnant smokers is more stressful than standard care. METHOD: Nine hundred eighteen U.K. women currently smoking on commencing antenatal care were randomized into three arms. Women in Arm A received one episode of brief advice to stop smoking. Women in Arm B were assessed for stage of change and worked through an exercise in self-help manuals on three occasions. Women in Arm C used a 20-min interactive computer program three times in addition to the intervention women in Arm B received. Stress was assessed by the change in score on the Perceived Stress Scale (PSS) from baseline to 30 weeks gestation, the month before delivery, and 10 days postpartum. RESULTS: There were small and not significant differences in the changes in PSS between the arms at all outcome times. There was no evidence that the importance women attached to pleasing their midwife by stopping, having failed to quit, or nulliparity modified the effect of intensive advice on change in stress levels. CONCLUSIONS.: Intensive advice to stop smoking was not associated with increases in stress. Advice and support for pregnant women to stop smoking should be given without fear of causing stress. PMID- 15749142 TI - Job stress and cardiovascular risk factors in male workers. AB - BACKGROUND: This study examined whether job stress (work demand and decision latitude) is associated with smoking, blood pressure, lipid level (total cholesterol, triglyceride, HDL cholesterol), and homocystein as risk factors for cardiovascular disease in Korean male workers. METHODS: Study subjects of this study were recruited from a sample of 1,071 workers in 20 companies of W city and H counties, and they were grouped into four categories (high strain group, active group, passive group, and low strain group) based on the postulation of Karasek's Job Strain Model. Of them, we invited 160 male workers (40 people each subgroup) using a stratified sampling, and finally, 152 eligible participants were analyzed. RESULTS: In multivariate analyses, we found that decision latitude was associated with cholesterol, triglyceride, and homocystein and that work demand was related to smoking and systolic blood pressure. Job strain (the combination of high work demand with low decision latitude) was significantly related to higher levels of homocystein after controlling for age, BMI, smoking, and social support at workplace. CONCLUSIONS: These results indicate that job stress is associated with cardiovascular risk factors and might contribute to the development of cardiovascular disease. Some considerations for the future research were discussed. PMID- 15749143 TI - Modulation of natural killer cell activity by supplementation of fermented milk containing Lactobacillus casei in habitual smokers. AB - BACKGROUND: Habitual smoking significantly reduces natural killer (NK) cell activity. To clarify whether the intake of fermented milk containing lactic acid bacteria restores NK cell activity in habitual smokers, we conducted a placebo controlled double-blind test. METHODS: Ninety-nine subjects with smoking habits were randomly divided into two groups and daily for 3 weeks were given fermented milk containing Lactobacillus casei or placebo. NK cell activity in the peripheral blood mononuclear cells (PBMNC) was determined before and after the intake. Average number of cigarettes smoked and number of cigarettes that the subjects smoked before giving blood after getting up on the inspection day were asked, and these data were used to adjust the influence of smoking on NK cell activity. RESULTS: NK cell activity in individuals was inversely correlated to numbers of cigarettes smoked. Averaged NK cell activity adjusted by the numbers of cigarettes was significantly higher in individuals drinking fermented milk containing L. casei than those drinking a placebo. However, the proportion of NK cells was not different between individuals drinking either fermented milk containing L. casei or the placebo. CONCLUSIONS: Intake of fermented milk containing lactic acid bacteria was considered effective for restoring the NK cell activity of habitual smokers. PMID- 15749144 TI - Prevention and health promotion in clinical practice: the views of general practitioners in Europe. AB - BACKGROUND: Disease prevention and health promotion are important tasks in the daily practice of all general practitioners (GPs). The objective of this study was to explore the knowledge and attitudes of European GPs in implementing evidence-based health promotion and disease prevention recommendations in primary care, to describe GPs' perceived barriers to implementing these recommendations and to assess how GPs' own health behaviors affect their work with their patients. METHODS: A postal multinational survey was carried out from June to December 2000 in a random sample of GPs listed from national colleges of each country. RESULTS: Eleven European countries participated in the study, giving a total of 2082 GPs. Although GPs believe they should advise preventive and health promotion activities, in practice, they are less likely to do so. About 56.02% of the GPs answered that carrying-out prevention and health promotion activities are difficult. The two most important barriers reported were heavy workload/lack of time and no reimbursement. Associations between personal health behaviour and attitudes to health promotion or activities in prevention were found. GPs who smoked felt less effective in helping patients to reduce tobacco consumption than non-smoking GPs (39.34% versus 48.18%, P < 0.01). GPs who exercised felt that they were more effective in helping patients to practice regular physical exercise than sedentary GPs (59.14% versus 49.70%, P < 0.01). CONCLUSIONS: Significant gaps between GP's knowledge and practices persist in the use of evidence-based recommendations for health promotion and disease prevention in primary care. PMID- 15749145 TI - Comparison of weight status among two cohorts of US Air Force recruits. AB - BACKGROUND: Overweight and obesity are increasing rapidly in the US and the Department of Defense (DoD). We examined whether weight trends evidenced in the general population and DoD are occurring among individuals entering US Air Force (USAF) Basic Military Training (BMT). METHODS: Individuals entering the USAF in 1996 (AF1996; N = 29,036) and 2000 (AF2000; N = 31,080), ages 17-29 years were surveyed. The two recruit cohorts were compared to age-matched individuals from the 1996 (N = 22,153) and 2000 (N = 31,861) Behavioral Risk Factor Surveillance System Survey (BRFSS). RESULTS: Crude rates for all age groups and age- and gender-standardized prevalence rates reflected significant increases in overweight/obesity among recruits. The direct standardized prevalence of overweight/obesity increased nearly 24%, from 14.8% in AF1996 to 18.3% in AF2000. The increase in overweight/obesity was particularly large among male recruits ages 25-29 (i.e., from 36.4% to 44.5%) between 1996 and 2000. CONCLUSIONS: USAF cohorts were less likely to be overweight than corresponding BRFSS samples. There were 19.1 and 20.2 percentage point differences between overall crude rates of overweight/obesity between AF1996 and BRFSS 1996 and AF2000 and BRFSS 2000, respectively. Nevertheless, overall rates of overweight and obesity are increasing among young recruits in the USAF at a fairly marked rate (approximately one percentage point per year). PMID- 15749146 TI - More than the loss of a library. PMID- 15749147 TI - Selling the silver: country house libraries and the history of science. AB - Country houses have long acted as scholarly research centres, and the contents and organization of their libraries reveal how knowledge was created and transmitted through scientific networks: they provide material evidence of intellectual and social cultures. Manuscripts are particularly appreciated in this regard because they are unique, but individual books also differ and copies of the same book are not always identical -- they might be annotated by authors and readers, and the printed text can vary as well -- so they also carry invaluable historical information. When libraries are broken up, access to the past is permanently blocked. PMID- 15749148 TI - The melancholy of anatomy. AB - In the middle of the 17th century, two founding fellows of the Royal Society -- John Evelyn and William Petty -- commissioned portraits of themselves with skulls. The paintings were commemorative, because Evelyn was celebrating his engagement and Petty had recently acquired an anatomical post at Oxford, but both sitters also intended their pictures to serve as reminders of mortality. PMID- 15749149 TI - Caroline Herschel: 'the unquiet heart'. AB - Caroline Herschel was famous in her own time as the discoverer of eight comets, but of even greater significance was the help she gave her brother William in his exploration of 'the construction of the heavens'. She acted as his amanuensis during night watches, wrote up neat copies of their observing records and prepared his papers for publication. She also compiled an index to John Flamsteed's Star Catalogue, which was published by the Royal Society at its own expense, and after William's death she reorganized his catalogues of nebulae so that his son John could revise his father's work. Yet Caroline's was a hard and largely loveless life, for which she found the recognition that came her way a scant consolation. PMID- 15749150 TI - Mary Anning: the fossilist as exegete. AB - The fossil hunter Mary Anning began collecting her 'curiosities' at a time when the age and nature of these relics from the past were little understood. Her spectacular discoveries of prehistoric marine reptiles, fossilized fish and a pterosaur touched off a geology-mania around the world. Two documents that have escaped previous analysis cast light on the religious journey of this remarkable woman. An eight-page manuscript at the Natural History Museum in London indicates a fascination with Benjamin West's painting 'Christ Rejected' and sophistication in Biblical interpretation; and a commonplace book at the Dorset County Museum in Dorchester tracks the shift in her Christian denomination from dissent to the Established Church, providing further indication of her spiritual depth - a piety that furthered, rather than hindered, her scientific progress. PMID- 15749151 TI - Carl Schmidt -- a chemical tourist in Victorian Britain. AB - Carl Schmidt is one of the founders of modern biochemistry. He wrote numerous scientific publications, and left future generations several important documents that included two extensive hand-written travel reports from 1857 and 1864. In these, Schmidt paints a lively and impressive picture of industry in Victorian Britain from a personal perspective, showing this key period of European economic history in a unique light. Thus, it is rewarding to follow Schmidt on his travels through Britain, which he regarded as the workshop of the world and a shining but unattainable example for the industrial development of his home country. PMID- 15749152 TI - Science in the 19th-century zoo. AB - The 19th century saw the advent of the modern zoological garden. The newly founded zoos not only claimed to educate and entertain their audiences, but also to serve science by providing direct access to exotic animals. However, reality did not live up to the promise of such rhetoric. The vast majority of biologists preferred to use dead bodies as the material for their morphological research. Nevertheless, there was still a strong interaction between the zoo and science. In the debate on Darwinism, the apes in the cage played a vital role. PMID- 15749153 TI - The rise, fall and resurrection of group selection. AB - The changing fate of group selection theory illustrates nicely the importance of studying the history of science. It was Charles Darwin that first used something like group selection to explain how natural selection could give rise to altruistic behavior and moral instinct. These instincts could be accommodated by his theory of evolution, he argued, if they had evolved 'for the good of the community'. By the 1960s, group selection had a new and vocal advocate in V.C. Wynne-Edwards. But this gave critics of the theory that selection might act on groups, rather than at the level of individuals or genes, a definable target, and from the mid-1960s to the 1980s group selection was considered the archetypal example of flawed evolutionary thinking. However, at the end of the 20th century ideas of group selection re-emerged as an important component of a multilevel theory of evolution. PMID- 15749154 TI - Mechanisms of platinum drug resistance. AB - Platinum-based drugs are among the most active anticancer agents available and are used widely for the treatment of a variety of human solid tumors. Although patients show high response rates to platinum drugs, most patients develop resistance to these drugs during treatment. Because the acquisition of resistance is a major obstacle to the clinical use of platinum drugs, the processes by which cells develop such resistance are of great interest and efforts have been made to overcome this problem. Both mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) cascades are involved in resistance to these drugs, and clinical trials of some small-molecule inhibitors of the MAPK and PI3K Akt cascades to overcome resistance to platinum drugs are ongoing. PMID- 15749155 TI - A possible common pharmacophore in the non-peptide antagonists of the bradykinin B1 receptor. PMID- 15749156 TI - Predicting drug-hERG channel interactions that cause acquired long QT syndrome. AB - Avoiding drug-induced cardiac arrhythmia is recognized as a major hurdle in the successful development of new drugs. The most common problem is acquired long QT syndrome caused by drugs that block human ether-a-go-go-related-gene (hERG) K(+) channels, delay cardiac repolarization and increase the risk of torsades de pointes arrhythmia (TdP). Not all hERG channel blockers induce TdP because they can also modulate other channels that counteract the hERG channel-mediated effect. However, hERG channel blockade is an important indicator of potential pro arrhythmic liability. The molecular determinants of hERG channel blockade have been defined using a site-directed mutagenesis approach. Combined with pharmacophore models, knowledge of the drug-binding site of hERG channels will facilitate in silico design efforts to discover drugs that are devoid of this rare, but potentially lethal, side-effect. PMID- 15749157 TI - Individual responder analyses for pain: does one pain scale fit all? AB - The outcomes of clinical trials are based on the mean responses of large numbers of subjects but fail to address inter-individual differences. The molecular mechanisms that underlie pain vary among individuals over time and among different types of pain to produce wide inter-individual variations in pain perception and response. Gender, ethnicity, temperament and genetic factors also contribute to individual variation in pain sensitivity and responses to analgesics. Pain measurement scales can be used differently across individuals based on the past pain experiences of individuals. We propose that individual responder analyses could be used in clinical trials to better detect analgesic activity across patient groups and within sub-groups, and to identify molecular genetic mechanisms that contribute to individual variation. PMID- 15749158 TI - Emerging role of homo- and heterodimerization in G-protein-coupled receptor biosynthesis and maturation. AB - The idea that G-protein-coupled receptors (GPCRs) can function as dimers is now generally accepted. Although an increasing amount of data suggests that dimers represent the basic signaling unit for most, if not all, members of this receptor family, GPCR dimerization might also be necessary to pass quality-control checkpoints of the biosynthetic pathway of GPCRs. To date, this hypothesis has been demonstrated unambiguously only for a small number of receptors that must form heterodimers to be exported properly to the plasma membrane (referred to as obligatory heterodimers). However, increasing evidence suggests that homodimerization might have a similar role in the receptor maturation process for many GPCRs. PMID- 15749159 TI - Interfacial inhibition of macromolecular interactions: nature's paradigm for drug discovery. AB - One of nature's strategies for interfering with molecular interactions is to trap macromolecules in transition states with their partners in dead-end complexes that are unable to complete their biological function. This type of inhibition, which we refer to as "interfacial inhibition", is illustrated by two natural inhibitors, brefeldin A (BFA) and camptothecin (CPT), whose modes of action have been elucidated fully in structural studies. Interfacial inhibition occurs at the protein-protein interface in the case of BFA and at the protein-DNA interface in the case of CPT. In both systems, the drugs take advantage of transient structural and energetic conditions created by the macromolecular complex, which give rise to "hot-spots" for drug binding. In addition to these examples, several natural compounds such as forskolin, tubulin inhibitors and immunophilins target protein interfaces. We propose that interfacial inhibition is a paradigm for the discovery of drugs that interfere with macromolecular complexes. PMID- 15749160 TI - Receptors coupled to heterotrimeric G proteins of the G12 family. AB - Much regarding the engagement of the G(12) family of heterotrimeric G proteins (G(12) and G(13)) by agonist-activated receptors remains unclear. For example, the identity of receptors that couple unequivocally to G(12) and G(13) and how signals are allocated among these and other G proteins remain open questions. Part of the problem in understanding signaling through G(12) and G(13) is that the activation of these G proteins is rarely demonstrated directly and is instead presumed usually from far removed downstream events. Furthermore, receptors that couple to G(12) and G(13) invariably couple to additional G proteins, and thus few events can be linked unambiguously to one G protein or another. In this article, we document receptors that reportedly couple to G(12), G(13) or both G(12) and G(13), evaluate the methodology used to understand the coupling of these receptors, and discuss the ability of these receptors to couple also to G(q). PMID- 15749161 TI - Angiotensin II, angiotensin II antagonists and spironolactone and their modulation of cardiac repolarization. AB - Angiotensin II and aldosterone produce pro-arrhythmic effects by several mechanisms, including the modulation of voltage-dependent K(+) channels involved in human cardiac repolarization. Drugs that inhibit the renin-angiotensin aldosterone system exert anti-arrhythmic actions that are related to the blockade of the pro-arrhythmic actions of angiotensin II and aldosterone. These anti arrhythmic actions include inhibition of electrical and structural cardiac remodeling, inhibition of neurohumoral activation, reduction of blood pressure and stabilization of electrolyte disturbances. In this article, several angiotensin II AT(1) receptor antagonists (candesartan, E3174, eprosartan, irbesartan and losartan) and aldosterone receptor antagonists (canrenoic acid and spironolactone) that directly modulate the activity of the voltage-dependent K(+) channels are reviewed; the effects of these antagonists might be useful in the prevention and treatment of cardiac arrhythmias. PMID- 15749162 TI - Endothelium-derived C-type natriuretic peptide: more than just a hyperpolarizing factor. AB - The perceived importance of C-type natriuretic peptide (CNP) in the mammalian vasculature has been raised by its recent identification as an endothelium derived hyperpolarizing factor (EDHF). This aspect of its biological activity is likely to be significant in the regulation of vascular tone, local blood flow and systemic blood pressure. However, the importance of CNP to cardiovascular homeostasis is likely to extend beyond that of a "hyperpolarizing factor" ; indeed, there is evidence that CNP has a key role in preventing smooth muscle proliferation, leukocyte recruitment and platelet reactivity. As such, endothelium-derived CNP is likely to exert a strong anti-atherogenic influence on blood vessel walls and represent a new therapeutic target in the fight against inflammatory cardiovascular disorders. Moreover, this profile of activity defines a new paradigm for the biological significance of EDHF. PMID- 15749163 TI - Why glucose transport in the brain matters for PET. AB - Neuronal activity is fueled by glucose metabolism, a phenomenon exploited in basic research and clinical diagnosis using fluorodeoxyglucose positron emission tomography (FDG-PET). According to the current view, glucose transport into the brain is not rate-limiting; thus, it cannot exert control over metabolism. This article challenges such a view by showing that basal transport hovers near its maximum, making metabolic activation unable to increase flux on its own. In the light of recent evidence on the identity of the cell type that preferentially breaks down glucose, we suggest that FDG-PET reports the synergistic activation of glucose transport and metabolism in astrocytes, rather than in neurons. PMID- 15749164 TI - Functional MRI using molecular imaging agents. AB - Contrast agents for magnetic resonance imaging (MRI) have recently been used as cellular-level probes of neural function. New in vivo labeling strategies now enable researchers to follow plasticity of brain activation patterns and cellular structure over time. On the horizon is the prospect that molecular imaging agents specifically designed for functional imaging (fMRI) on a relatively fast timescale could offer an alternative to conventional hemodynamics-based approaches. Development of several MRI sensors has defined principles by which imaging agents for "molecular fMRI" can be constructed; application of engineered sensors for cellular-level correlates of neuronal activity would allow researchers to combine the noninvasiveness of MRI with spatial resolution of tens of microns and temporal resolution of 100ms or less. Facilitated by advances in imaging-agent delivery methods and model systems appropriate for high-resolution neuroimaging, novel molecular imaging strategies continue to potentiate MRI as a tool for mechanistic investigation of neural systems. PMID- 15749165 TI - Song learning in birds: diversity and plasticity, opportunities and challenges. AB - A common trend in neuroscience is convergence on selected model systems. Underlying this approach is an often implicit assumption that mechanisms observed in one species are characteristic of all related species. Although the model system approach has been extremely productive, it might not account for all of the mechanistic differences between species that differ behaviourally. Using the neural system that regulates song learning in songbirds as an example, we demonstrate how integrating model system and comparative approaches can lead to a more complete picture of neural mechanisms, and can resolve issues raised by a focus on selected species. PMID- 15749166 TI - Surface trafficking of receptors between synaptic and extrasynaptic membranes: and yet they do move! AB - Concentration of neurotransmitter receptors at synapses is thought to result from stable binding to subsynaptic scaffold proteins. Recent data on synaptic plasticity have shown that changes in synaptic strength derive partly from modification of postsynaptic receptor numbers. This has led to the notion of receptor trafficking into and out of synapses. The proposed underlying mechanisms have under-evaluated the role of extrasynaptic receptors. Recent technological advances have allowed imaging of receptor movements at the single-molecule level, and these experiments demonstrate that receptors switch at unexpected rates between extrasynaptic and synaptic localizations by lateral diffusion. Variation in receptor numbers at postsynaptic sites is therefore likely to depend on regulation of diffusion by modification of the structure of the membrane and/or by transient interactions with scaffolding proteins. This review is part of the TINS Synaptic Connectivity series. PMID- 15749167 TI - Fishing for key players in mechanotransduction. AB - The senses of touch and hearing involve transduction of mechanical stimuli into electrical signals. The search for components of the transduction apparatus in mechanosensory neurons has benefited greatly from genetic approaches using both invertebrates and vertebrates. A consensus model has emerged that includes extracellular and intracellular structural components arranged around a central mechanically gated channel. In the sensory hair cell of the inner ear, the extracellular structural component thought to have a key role in opening the transduction channel is the tip link. Although elusive for decades, recent studies have yielded candidates for both the transduction channel and the tip link in hair cells. PMID- 15749168 TI - Orchestrating time: arrangements of the brain circadian clock. AB - Daily oscillations in physiology and behavior are regulated by a brain clock located in the suprachiasmatic nucleus (SCN). Individual cells within this nucleus contain an autonomous molecular clock. Recent discoveries that make use of new molecular and genetic data and tools highlight the conclusion that the SCN is a heterogeneous network of functionally and phenotypically differentiated cells. Neurons within SCN subregions serve distinctly separate functions in regulating the overall activity of the circadian clock: some cells within the SCN rhythmically express "clock" genes, whereas others exhibit induced expression of these genes after the organism has been exposed to a light pulse. The coordinated interaction of these functionally distinct cells is integral to the coherent functioning of the brain clock. PMID- 15749169 TI - The hypothalamic integrator for circadian rhythms. AB - Although the suprachiasmatic nucleus (SCN) is well established as providing a genetically based clock for timing circadian rhythms, the mechanisms by which the timing signal is translated into circadian rhythms of behavior and underlying physiology have only recently come to light. The bulk of the SCN outflow terminates in a column of tissue that arches upward and backward from the SCN, and which includes the subparaventricular zone (SPZ) and the dorsomedial nucleus of the hypothalamus. Neurons within the dorsal SPZ are necessary for organizing circadian rhythms of body temperature, whereas neurons in the ventral SPZ are needed for circadian rhythms of sleep and waking. Ventral SPZ neurons in turn relay to the dorsomedial nucleus, which is crucial for producing circadian rhythms of sleep and waking, locomotor activity, feeding and corticosteroid production. This multistage processor provides the animal with flexibility so that environmental cues, such as food availability, ambient temperature and social interactions, can be integrated with the clock signal to sculpt an adaptive pattern of rhythmic daily activities that maximize the chances of survival and reproduction. PMID- 15749170 TI - Space matters: local and global dendritic Ca2+ compartmentalization in cortical interneurons. AB - Dendrites of pyramidal neurons are complex, electrically active structures that can produce local and global Ca(2+) compartments. Recent studies indicate that dendrites of cortical GABAergic interneurons are also highly specialized, and that different subtypes vary in their morphology, in their intrinsic and synaptic conductances and in the Ca(2+) signals they generate. Because interneurons play a major role in oscillations, understanding their dendrites could offer key insights into rhythmogenesis. Different interneuron subtypes have different synaptic integration properties and generate differentially timed inhibition at distinct sites of the pyramidal neuraxis. In addition, interneuron dendrites generate diverse Ca(2+) signals that reflect this circuit function and probably also implement subclass-specific plasticity and homeostasis. PMID- 15749171 TI - Exercise and the endothelial cell. AB - Regular exercise is known to be effective in the prevention and treatment of cardiovascular disease. Among the cardioprotectant mechanisms influenced by exercise, the endothelium is becoming recognised as a major target. Preservation of endothelial cell structure is vital for frictionless blood flow, prevention of macrophage and lipid infiltration and, ultimately, optimal vascular function. Exercise causes various kinds of mechanical, chemical and thermal stresses, and repeated exposure to these stresses may precondition the endothelial cell to future stresses through a number of different mechanisms. This review discusses stress-induced changes in endothelial cell morphology, biochemistry and components of platelet activation and cell adhesion that impact on endothelial cell structure. An enhanced understanding of the effects of exercise on the endothelial cell will assist in directing future research into the prevention of cardiovascular disease. PMID- 15749172 TI - Positive pressure ventilation in the management of acute and chronic cardiac failure: a systematic review and meta-analysis. AB - Chronic heart failure (CHF) is a common condition and is associated with excess morbidity and mortality, in spite of the many advances in its treatment. Chronic stable heart failure is also associated with an increased incidence of sleep related breathing disorders, such as central sleep apnoea (CSA) and Cheyne Stokes respiration (CSR). Continuous positive airways pressure (CPAP) has been shown to alleviate the symptoms of CHF, improve left ventricular function and oxygenation. To a certain extent, CPAP also abolishes sleep-related breathing disorders in patients with chronic heart failure. In patients with acute pulmonary oedema, the use of positive pressure ventilation improves cardiac haemodynamic indices, as well as symptoms and oxygenation, and is associated with a lower need for intubation. However, some studies have cast doubts about its safety and suggest a higher rate of myocardial infarction associated with its use. In our opinion, non invasive positive pressure ventilation and CPAP offers an adjunctive mode of therapy in patients with acute pulmonary oedema and chronic heart failure, who may not be suitable for intubation and in those not responsive to conventional therapies. Non-invasive ventilation also helps to improve oxygenation in those patients with exhaustion and respiratory acidosis. Many trials are still ongoing and the results of these studies would throw more light on the present role of non-invasive ventilation in the management of CHF. PMID- 15749173 TI - Development of indications for cardiac resynchronisation therapy in the implantable cardioverter defibrillator population. PMID- 15749174 TI - Sixteen-row multislice computed tomography in infants with double aortic arch. AB - BACKGROUND: The introduction of multislice computed tomography (MSCT) in 1998 has led to a considerable boost of CT angiography. Four-row MSCT has been employed in the diagnostic assessment of vascular rings. Sixteen-row MSCT is expected to further increase the diagnostic power of MSCT. We report three cases of double aortic arch diagnosed by a 16-row MSCT, and discuss the possible advantages of this diagnostic tool. PATIENTS AND METHODS: From April 1, 2003, to September 1, 2003, three patients underwent 16-row MSCT at our institution to evaluate the possible presence of a vascular ring. All patients presented with stridor and feeding difficulties. MSCT was performed under bland sedation (chloral hydrate 60 mg/kg). ECG gating and breath hold were not employed. Three-dimensional reconstructions were employed to assess the presence of airway compromise as well as of vascular anomalies. RESULTS: Scanning time averaged 4 s (3.8-4.5 s). Motion artefacts were not relevant. The CT scan showed the existence of a vascular anomaly as well as of a significant tracheal compression in all cases, and was considered conclusively diagnostic by the cardiac surgeons. All three patients underwent uncomplicated corrective surgery without further investigations. CONCLUSION: Sixteen-row MSCT with 3D reconstruction allows a precise evaluation of the airway compromise and a detailed assessment of the anatomy of vascular anomalies. The very short scanning time allows to avoid deep sedation and anaesthesia, and to obtain higher image quality and spatial resolution, with a concomitant reduction of the radiation dose. PMID- 15749175 TI - Superiority of surgical versus medical treatment in patients with Staphylococcus aureus infective endocarditis. AB - BACKGROUND: We present here the clinical features and outcome of 54 patients affected by a Staphylococcus aureus infective endocarditis at the Amiens hospital between 1990 and 2000. The patients operated-on, group A (20 patients), were compared to the population of patients treated by exclusive antibiotherapy, group B (34 patients). PATIENTS AND METHOD: The male gender predominated with a sex ratio of 2.6. The mean age of the global population was 58.7+/-1.6 years. Time between onset of endocarditis symptoms and treatment (entire group) ranged from 1 to 120 days (mean 14.4 days). The main portal of entry were, respectively, for group A and group B: cutaneous 55% and 44.1%; intravascular material 5% and 8.8%; and rhinopharynx 5% and 8.8%. Seventy-five percent of the Staphylococcus aureus isolated were Methi-S. The main surgical treatment indication were: hemodynamic failure (HF) (30%), unstable infection with collapse (UI) (30%), UI+HF (10%), voluminous vegetation (20%) and embolism event (10%). RESULTS: The hospital mortality rate were respectively for the entire group, group A and group B: 25%, 35% and to 41% (ns). For group A, the operative mortality was lower(21%) after the first week. The actuarial survival rate (Kaplan-Meier) after 24 months was 54./+/-6.9% for the global population and 74+/-10.6% for group A and 43+/-8.5 for group B (p<0.001). The multivariate analysis finds severe sepsis and index of comorbidity as independent factors related to the global late mortality and, respectively, the age and the severe sepsis for group A, and the cardiac insufficiency for group B. CONCLUSION: The surgical treatment seems to be the best way to improve the results after Staphylococcus aureus endocarditis. The severity of the sepsis remains the most severe prognostic element, whatever the treatment adopted may be. PMID- 15749176 TI - Long-term outcomes after treatment of diffuse in-stent restenosis with rotational atherectomy followed by beta-radiation therapy with a 188Re-MAG3-filled balloon. AB - BACKGROUND: Intracoronary radiation therapy for in-stent restenosis has been demonstrated to reduce restenosis and major adverse cardiac events. However, long term angiographic and clinical outcomes after beta radiation therapy have not been sufficiently evaluated. METHODS: We evaluated the long-term angiographic and clinical outcomes of 50 consecutive patients who had received beta-radiation therapy with a 188Re-MAG3-filled balloon after rotational atherectomy for diffuse in-stent restenosis (lesion length>10 mm) in native coronary arteries. The radiation dose was 15 Gy at a depth of 1.0 mm into the vessel wall. RESULTS: The mean lesion length was 25.6+/-12.7 mm. Radiation was delivered successfully to all patients without any procedural or in-hospital complications. At the 6-month angiogram, the restenosis rates was 10% (5/50). There were no major adverse cardiac events (MACE), such as death, myocardial infarction, and target lesion revascularization (TLR) by 6-month follow-up. Long-term clinical follow-up data were obtained in all patients during 30.1+/-4.5 months. No myocardial infarction and one noncardiac death occurred during follow-up. Two-year follow-up angiogram was performed in 26 (58%) of 45 patients who showed a patent radiation segment at the 6-month angiogram. Significant narrowing of diameter stenosis of more than 50% occurred in 6 (23%) of 26 patients between 6 and 24 months after beta radiation. Late TLR was performed in 6 patients. The rate of 30-month death-free survival and MACE-free survival were 98.0+/-2.0% and 86.9+/-5.0%. CONCLUSION: Beta-radiation using a 188Re-MAG3-filled balloon after rotational atherectomy is associated with favorable long-term angiographic and clinical outcomes. PMID- 15749177 TI - Cardiac troponin I release in acute pulmonary embolism in relation to the duration of symptoms. AB - PURPOSE: To evaluate the release of cardiac troponin I in normotensive patients with acute pulmonary embolism in relation to the duration of symptoms. METHODS: Fifty-seven normotensive patients with acute pulmonary embolism were included in the study. Patients were divided into two groups based on the duration of symptoms at presentation: symptoms of < or =72 h, group A; symptoms of >72 h, group B. Serum cardiac troponin I levels were measured at presentation. RESULTS: Mean age was 63+/-18 years and 23 (40%) patients were males. Thirty-three (58%) patients had symptoms of < or =72 h (group A) and 24 (42%) had symptoms of >72 h (group B). Both groups had similar prevalence of right ventricular dysfunction on echocardiography (55% [n=18] in group A vs. 42% [n=10] in group B, p=NS). Sixteen patients had elevated serum cardiac troponin I (mean+/-S.D. 3.3+/-2.3 ng/ml, range 0.6-8.3 ng/ml). Elevated serum cardiac troponin I was strongly associated with right ventricular dysfunction (p=0.015). All patients with elevated serum cardiac troponin I (n=16) were in group A (p<0.0001). Twelve of 18 (67%) patients with (p=0.0005) and 4 of 15 (27%) patients without (p=NS) right ventricular dysfunction had elevated serum cardiac troponin I. Thirteen of 16 (81%) patients with elevated serum cardiac troponin I had duration of symptoms < or =24 h at presentation. CONCLUSIONS: The dynamics of cardiac troponin I release in acute pulmonary embolism in patients who present with symptoms of < or =72 h duration could be different from those who present with longer duration of symptoms. Therefore, the use of cardiac troponin I in risk stratification of acute pulmonary embolism might be limited to the patients presenting within 72 h of the onset of symptoms. PMID- 15749179 TI - Influence of etiology on ventilatory expired gas and prognosis in heart failure. AB - BACKGROUND: Mechanisms leading to heart failure (HF) are numerous. Etiology-based differences are, however, frequently not taken into account when assessing results of an exercise test. The purpose of this investigation is to: (1) compare subject characteristics and ventilatory expired gas measures demonstrating clinical value between subjects with ischemic and non-ischemic HF. (2) Examine the prognostic value of HF etiology. METHODS: 71 subjects (44 male/27 female) diagnosed with compensated HF underwent exercise testing with ventilatory expired gas analysis. Mean age and ejection fraction (EF) were 51.3% (+/-12.8) and 27.0% (+/-12.5%) for the entire group. HF etiology was ischemic in 30 subjects (16 male/14 female) and non-ischemic in 41 (28 male/13 female). RESULTS: Age (57.5+/ 10.7 vs. 46.7+/-12.5, p<0.001), minute ventilation-carbon dioxide production (VE/VCO2) slope (39.5+/-9.1 vs. 32.6+/-7.7, p=0.001), and duration of phase 1 kinetics in seconds (46.0+/-23.0 vs. 30.0+/-15.6, p=0.001) were significantly higher while peak oxygen consumption (VO2) in ml O2 kg(-1) min(-1) (12.5+/-4.7 vs. 16.0+/-5.2, p=0.006) and partial pressure of end-tidal CO2 (P(ET)CO2) in mm Hg at rest (32.1+/-4.8 vs. 36.1+/-8.0, p=0.02) and peak exercise (31.7+/-4.3 vs. 36.2+/-5.9, p=0.001) were significantly lower in the ischemic group. Difference in EF did not reach statistical significance (28.4% +/-12.5%-ischemic vs. 26.1%+/ 12.5%-non-ischemic, p=0.44). Ischemic etiology was additionally a significant predictor of cardiac-related events (p=0.04). CONCLUSIONS: These findings demonstrate noninvasive indicators of cardiac function and prognosis is poorer in subjects with ischemic HF etiology. Consideration of HF etiology may therefore be prudent, particularly when considering prognosis. PMID- 15749178 TI - Acute pulmonary embolism in elderly: clinical characteristics and outcome. AB - OBJECTIVE: To evaluate the clinical characteristics and outcome of acute pulmonary embolism in elderly in comparison to the younger patients. METHODS: Study population consisted of 136 patients with a confirmed diagnosis of acute pulmonary embolism. Clinical characteristics and thromboembolic risk factors were analyzed between the elderly (> or =65 years of age) and the younger (<65 years of age) patients. In-hospital mortality was used as a measure of outcome. RESULTS: Elderly group consisted of 70 patients (age 76.4+/-8.3 years, range 65 96 years; females 58%) and younger group of 66 patients (age 48.5+/-12 years, range 18-64 years, females 59%). Syncope was more frequent in elderly group (19% vs. 6%, P=0.03) but the symptoms of shortness of breath and pleuritic chest pain were not significantly different between groups. Malignancy was the most common risk factor for thrombo-embolism, but immobilization predominated among patients in elderly group (21% vs. 6%, P=0.01). Tachycardia was common in younger patients compared to the elderly. Ventilation-perfusion scan was used more commonly in younger patients (76% vs. 57%, P=0.02), whereas, helical computed-tomography scan was used equally in both groups. Most of the patients had lower extremity duplex study (97% in each group). Inferior vena cava filter placement was common and thrombolytic therapy rare among elderly patients. Patients in elderly group had higher in-hospital mortality (17% vs. 5%, P=0.02). CONCLUSIONS: Syncope is a more frequent presenting symptom and immobilization a common risk factor in elderly patients with acute pulmonary embolism. In addition, they have higher in-hospital mortality. PMID- 15749180 TI - Plasma thioredoxin levels in patients with unstable angina. AB - Plasma levels of human thioredoxin are indicative of the responses against oxidative stress. We measured the plasma thioredoxin levels in patients with unstable angina in order to examine the relationships between subsequent clinical course and plasma thioredoxin levels before and after treatment for unstable angina. Blood was sampled both on admission and after treatment in 44 patients with unstable angina. In addition, blood samples were obtained from 41 patients with stable exertional angina and 41 patients with chest pain syndrome after admission. The plasma levels of thioredoxin were the highest in the unstable angina group among three groups (p<0.001). Treatment of unstable angina decreased the plasma thioredoxin levels (p<0.01). We divided the patients with unstable angina into two groups according to the plasma thioredoxin levels on admission and after treatment. There was a significant difference in Braunwald's classification between the high thioredoxin and the low thioredoxin group on admission, as analyzed by the chi2 test with Yates's correction (p<0.05). Moreover, there was a significant difference in incidence of recurrent anginal attacks at rest between the high thioredoxin and the low thioredoxin group after treatment, as analyzed by the chi2 test with Yates's correction (p<0.001). The present study demonstrated that plasma thioredoxin levels are significantly increased in patients with unstable angina compared to those with stable exertional angina and chest pain syndrome. Thioredoxin levels were associated with recurrent myocardial ischemia in patients with unstable angina. PMID- 15749181 TI - Myocardiocyte apoptosis in heart failure in chronic Chagas' disease. AB - Chagas' disease is caused by the parasite Trypanosoma cruzi. The disease affects 16-18 million patients in South America and heart involvement is the major cause of morbidity and mortality of the disease. The myocarditis observed during the chronic phase affects patients independently of the clinical manifestation, although patients with heart failure present an intense degree of myocarditis and fibrosis. To address the pathogenesis of heart failure in Chagas' disease, we investigated the role of myocardial cell loss by apoptosis in patients in the chronic phase of Chagas' disease. Apoptosis was also evaluated in inflammatory cells. Twenty-two specimens of the left ventricle were obtained during autopsies. Eleven samples from patients with heart failure and equal number from patients without heart failure. The material was analyzed by TUNEL methods to identify early apoptotic events and fibrosis was evaluated on HE-stained slides. In patients with heart failure, the extent of fibrosis and the number of apoptotic myocardial and inflammatory cells were significantly higher than in specimens obtained from patients without heart failure. These results suggest that myocardial cell loss by apoptosis and fibrosis contribute to heart failure in the chronic phase of Chagas' disease. PMID- 15749182 TI - Anthropometric variables and physical activity as predictors of cardiac cachexia. AB - BACKGROUND: This study explored the frequency of cardiac cachexia in Mexican patients, the role of anthropometric variables as predictors of its development and its association with food intake and physical activity. METHODS: Seventy three patients with systolic heart failure were included in the study. Cardiac cachexia was defined as weight loss of >6.0% in 6 months. Anthropometric data, physical activity and dietary intake were evaluated by a 3-day questionnaire at the beginning of the study and 6 months later. RESULTS: After 6 months of follow up, 14 (19%) patients developed cachexia with a mean weight loss of 12.1+/-3.4%. Significant decrease in the anthropometric variables were observed in patients which developed cachexia except in the waist to hip ratio, which increased in these patients due to bigger diminish of hip circumference than in the waist one. The subjects which developed cachexia had significant less physical activity after 6 months (-6.9%) in comparison with the non cachexic group. Reported energy intake did no differ among groups. Patients with cardiac cachexia showed greater prevalence of obesity and overweight, a high body fat percentage and a low arm circumference. CONCLUSIONS: Cardiac cachexia development was not related with low energy intake or increase in the total energy expenditure (explained by the physical activity). The only variable related to cachexia development was lower physical activity. PMID- 15749183 TI - The prognostic value of troponin T and echocardiography in acute pulmonary edema. PMID- 15749184 TI - Minor myocardial damage detected by troponin T is a powerful predictor of long term prognosis in patients with acute decompensated heart failure. AB - BACKGROUND: The progression of chronic heart failure (CHF) is characterized by frequent exacerbation requiring hospitalization and high mortality. Clinical deterioration is triggered by many factors that could promote ongoing myocytes injury. We sought to determine whether a specific marker of cardiac injury, troponin T (cTnT), is associated with prognosis in acute decompensated heart failure (ADHF). METHODS: One hundred and eighty-four consecutive patients with ADHF were enrolled in the absence of an acute coronary syndrome. A cTnT value> or =0.1 ng/ml in samples drawn at 6, 12 or 24 h after hospital admission was considered abnormal. RESULTS: Increased levels of cTnT were found in 58 patients (31.5%, group 1). There were no significant differences between group 1 and patients with cTnT<0.1 ng/ml (group 2) in terms of demographic and clinical characteristics, although ischemic etiology was more prevalent in group 1 (51.7% vs. 31.7%, p=0.009). During follow-up, the mortality in groups 1 and 2 was 31% and 17.5% (p=0.038, OR=2.13, 95% CI: 1.03-4.69), respectively. The 3-year free CHF readmission survival in group 1 and 2 was 25% and 53% (log rank test p=0.015). In a Cox proportional hazard model, poor tissue perfusion (HR=2.46, 95% CI=1.31-4.6), previous infarction (HR=1.99, 95% CI=1.02-3.9) and cTnT> or =0.1 ng/ml (HR=1.74, 95% CI=1.05-2.9) emerged as the independent predictors of long term outcome. CONCLUSIONS: One third of patients with decompensated CHF had elevated levels of cTnT. Troponin T was an independent long-term prognostic marker of morbidity and mortality and it suggests a role of biochemical risk stratification in this setting. PMID- 15749185 TI - A study of lipoprotein lipase gene intron 8 polymorphisms in Chinese Han race essential hypertension patients. AB - OBJECTIVE: To investigate the association of lipoprotein lipase gene intron 8 polymorphisms and Essential Hypertension in Han race Chinese. METHODS AND RESULTS: 116 patients with Essential Hypertension were enrolled and another 116 normal people were served as controls. All cases were examined for the genotypes of intron 8 in lipoprotein lipase gene by the methods of polymerase chain reaction restriction fragment length polymorphism, and the serum lipoprotein levels were also observed. Results showed that body mass index blood pressure and the serum triglyceride level were obviously increased in the Essential Hypertension group. The genotype and allele frequency of intron 8 in lipoprotein lipase in the Essential Hypertension group showed obvious differences compared with the control group. Serum triglyceride levels were higher in the patients with H+H+ genotype than in those in non H+H+ genotype of intron 8 in lipoprotein lipase by HindIII digestion. The systolic blood pressure showed a decreasing tendency among the H+H+ genotype, the H+H- genotype and the H-H- genotype individuals. CONCLUSION: The results suggest that lipoprotein lipase may be an important genetic factor associated with the Chinese Han race Essential Hypertension patients. The polymorphisms of intron 8 in lipoprotein lipase influence the blood-lipid metabolism, induce blood vessel rebuilding and play an important role in the invasion and development of Essential Hypertension. PMID- 15749186 TI - Risk factors for first-ever acute ischemic non-embolic stroke in elderly individuals. AB - BACKGROUND: Stroke is a leading cause of mortality and subsequent serious long term physical and mental disability among survivors. In the elderly, ischemic stroke accounts for more than 80% of all strokes. OBJECTIVES: To identify major risk factors for a first-ever acute ischemic/non-embolic stroke in individuals older than 70 years. METHODS: A population-based case-control study of patients admitted to the University Hospital of Ioannina, Epirus, Greece, due to first ever ischemic/non-embolic stroke from March 1997 to January 2002. All patients were subjected to brain CT and had their serum lipids and biochemical metabolic parameters determined within 24 h from the onset of symptoms. RESULTS: A total of 163 (aged>70 years) consecutive stroke patients and 166 apparently healthy volunteers were studied. An atherogenic lipid profile and metabolic disturbances were more prevalent in the patient group than in stroke-free controls. Multivariate logistic regression analysis identified diabetes mellitus (odds ratio (OR), 1.92; 95% CI, 1.02-3.63), triglycerides (TG) (OR, 1.16; 95% CI, 1.09 1.22), HDL-cholesterol (OR, 0.57; 95% CI, 0.43-0.76), apo A-I (OR, 0.80; 95% CI, 0.70-0.92), lipoprotein(a) [LP(a)] (OR, 1.51; 95% CI, 1.25-1.79), uric acid (OR, 1.30; 95% CI, 1.06-1.59) albumin (OR, 0.38; 95% CI, 0.20-0.70) fibrinogen (OR, 1.10; 95% CI, 1.05-1.13) and the metabolic syndrome (OR 2.48, 95% CI, 1.16-5.29) as significantly associated with ischemic/non-embolic stroke. CONCLUSION: Ischemic non-embolic stroke in the elderly is associated with dyslipidemia and several predictor metabolic factors, which could be substantially modified by lifestyle changes and therapeutic intervention. PMID- 15749187 TI - Myocardial disarray: an architectural disorganization linked with adrenergic stress? AB - BACKGROUND: Myocardial disarray is a structural abnormality found in specific zones of the normal heart. In some conditions, such as hypertrophic cardiomyopathy (HCM), its occurrence represents a pathological process leading to myocardial asynergy. The incidence of "pathological" myocardial disarray in humans is still not known. It has been suggested that a link exists between adrenergic overactivity and myocardial disarray. The aim of the present study is to compare heart findings in conditions with and without chronic sympathetic overtone for evidence of possible linkage in humans. MATERIALS AND METHODS: A total of 340 hearts were studied. They were divided into seven groups: sudden/unexpected coronary death; sudden/unexpected death in silent Chagas' disease; brain haemorrhage following berry aneurysm rupture; transplanted hearts; congestive heart failure, AIDS and cocaine abuse. Findings in these hearts were compared with anatomic changes in 92 control hearts, where the decedent had died from head trauma, electrocution, or carbon monoxide intoxication. The frequency and presence of myocardial disarray were recorded and correlated to heart weight, extent of myocardial fibrosis, and contraction band necrosis (CBN). RESULTS: Hearts from patients with conditions that increased sympathetic tone showed an association of myocardial disarray and contraction band necrosis without any relationship to heart weight. CONCLUSIONS: Myocardial disarray was observed in cardiac areas where it is not found normally. It was associated with adrenergic myocardial stress morphologically expressed by a higher number of foci (p<0.01) and myocells (p<0.001) with CBN versus findings in normal subjects. The condition deserves further study as a possible myocardial asynergic and arrhythmogenic factor especially in sudden/unexpected death. PMID- 15749188 TI - Ibutilide in persistent atrial fibrillation refractory to conventional cardioversion methods. AB - BACKGROUND: Electrical cardioversion of atrial fibrillation seems to be enhanced by pretreatment with ibutilide, but only few is known about the effects of ibutilide in atrial fibrillation which failed to convert with class III antiarrhythmic agents and electrical cardioversion. The objectives of this study were to evaluate the efficacy and safety of ibutilide administration in patients with persistent atrial fibrillation refractory to long-term therapy with class III antiarrhythmic drugs and transthoracic cardioversion. METHODS: Prospective study in 22 patients (16 men and 6 women, mean age 63+/-9 years) with structural heart disease and persistent atrial fibrillation for a mean duration of 39+/-50 (range 1-145) months. All patients had failed to convert to sinus rhythm after transthoracic cardioversion while on treatment with class III antiarrhythmic drugs (amiodarone in 82%, sotalol in 18%). One milligram of ibutilide was administered in all patients and electrical cardioversion was performed again, if necessary. RESULTS: The total conversion rate to sinus rhythm was 95% (21 of 22 patients). Two patients (9%) were successfully converted after ibutilide alone and 19 patients (86%) when transthoracic cardioversion was repeated after ibutilide. The QTc intervals increased from 451+/-28 to 491+/-49 ms (p<0.001) after ibutilide. No adverse effects occurred. The rate of freedom from atrial fibrillation after 1 month of follow-up was 64%. CONCLUSIONS: The efficacy of concomitant use of ibutilide infusion and, if necessary, repeated transthoracic cardioversion for restoration of sinus rhythm in long-term persistent atrial fibrillation and previously failed antiarrhythmic and electrical cardioversion was 95%. There were no adverse effects associated with ibutilde administration. Our results suggest that this combined strategy may be safe and successful in patients with atrial fibrillation resistant to conventional cardioversion methods and may be an alternative to internal cardioversion. PMID- 15749189 TI - Augmentation index, pulse pressure amplification and superoxide anion production in patients with coronary artery disease. AB - BACKGROUND: Free oxygen radicals appear to be involved in several processes that contribute to atherogenesis and increased arterial stiffness. METHODS: The aim of our study was to evaluate arterial stiffness and the production of superoxide anions by activated polymorphonuclear neutrophils (PMN) obtained from patients with stable coronary artery disease (CAD). Thirty four consecutive patients were studied (21 men, 13 women, mean age 58 years) who underwent coronary angiography. Arterial stiffness was assessed by pulse wave analysis using a validated system (Sphygmocor Mx, AtCor Medical). Superoxide anion production by activated neutrophils was determined by a spectrophotometric method involving the measurement of cytochrome C reduction. The extent of coronary narrowing was estimated by calculation of the Gensini score. RESULTS: Superoxide anion production by stimulated PMN showed a significant positive correlation with the augmentation index (AIx) and a significant negative correlation with pulse pressure amplification (PPA), (r=0.4, p=0.02; r=-0.5 and p=0.0026 respectively). In multivariable analyses, after adjustment for age, gender and Gensini score, superoxide anions and BMI were significant predictors of AIx (R2=57.37%, p=0.001) and PPA (R2=49.04%, p=0.008). Superoxide anion production was significantly higher in the middle (52.0+/-5.8 nmol O2-/2.5x10(6) PMN/30 min) and upper teriles (62.7+/-5.6) of AIx in comparison with the first tertile 31.8+/-4.1 (p< or =0.05, p< or =0.001). Moreover, superoxide anion production in the highest tertile of PPA was significantly lower (35.6+/-4.3 nmol O2-/2.5x10(6) PMN/30 min) than that in the tertile (60.8+/-6.2, p< or =0.05). Neither the augmentation index nor pulse pressure amplification correlate with the severity of coronary atherosclerosis as indicated by the Gensini score. CONCLUSIONS: markers of arterial stiffness, AIx and pulse pressure amplification correlate with superoxide anion production but not with the severity of atherosclerosis in coronary arteries. PMID- 15749191 TI - Neurohormones as markers of right- and left-sided cardiac dimensions and function in patients with untreated chronic heart failure. AB - BACKGROUND: It is now well accepted that neuroendocrine activation is of pathophysiological and prognostic importance in patients with chronic heart failure (CHF). We hypothesized that the different neuroendocrine factors reflect different aspects of the cardiac dysfunction in CHF patients and that neuroendocrine profiling could be of value. In order to study this, we investigated the relationship between hormones and cardiac dimensions and function of both the right and left ventricle. METHODS: Twenty-three patients with newly diagnosed, untreated CHF were included. Right (RVEF) and left ventricular ejection fractions (LVEF) and volumes were measured by means of first pass and equilibrium radionuclide ventriculography. RESULTS: LVEF was 0.29 (range: 0.11-0.55). Two-thirds of the patients had dilated left ventricles with volumes above upper reference limit. Right ventricular ejection fraction was normal in all subjects as well as right ventricular volumes. Likewise, on average, the lung transit time (LTT) was normal. Brain natriuretic peptide (BNP) significantly correlated with LVEF, left ventricular end-diastolic volume index (LVEDVI) and left ventricular end-systolic volume index (LVESVI). Adrenaline correlated significantly with both right ventricular end-diastolic volume index and right ventricular end-systolic volume index. Lung transit time correlated with atrial natriuretic peptide (ANP) and BNP (only ANP in multivariate analysis). CONCLUSIONS: (1) BNP reflects the LVEF as well as diastolic and systolic dimensions; (2) adrenaline reflects the right ventricular systolic and diastolic dimensions; and (3) ANP reflects the lung transit time. We conclude that "neuroendocrine profiling" may potentially be of diagnostic and therapeutic use. PMID- 15749190 TI - Vascular dysfunction after coarctation repair is related to the age at surgery. AB - BACKGROUND: Despite repair of aortic coarctation, hypertension is frequent in adults and premature coronary and cerebrovascular disease remain of concern. Persistent impairment of arterial dilation has been suspected to contribute to abnormal blood pressure regulation. We tested the hypothesis that arterial reactivity is more likely to be impaired in patients corrected at older age. METHODS: We studied changes in brachial artery diameter in response to reactive hyperemia (FMD) and to nitroglycerin (NMD) in 36 patients and 25 controls. Depending on their age at surgery, patients were divided in group A (surgery <9 years) and group B (surgery > or =9 years). RESULTS: Cholesterol levels and percentage of smokers were similar in patients and controls, but 16 patients had arterial hypertension compared to none of the controls. Endothelium-dependent vasodilation, FMD, and endothelium-independent vasodilation, NMD, were significantly impaired in patients vs. controls (8.2+/-6.2% vs. 13.0+/-5.1%, p<0.001 and 12.9+/-8.0% vs. 18.8+/-9.2%, p<0.01, respectively), both, in hypertensives (8.3+/-6.0%, p<0.01 and 11.8+/-6.0%, p<0.05) and in normotensives (8.1+/-6.5% p<0.01 and 13.8+/-9.3%, p<0.05). However, FMD and NMD in patients of group A did not significantly differ from that in controls (10.0+/-6.7% n.s. and 15.0+/-7.6% n.s.), whereas they were lowest in patients of group B (5.5+/-4.3%, p<0.0001 and 9.6+/-7.7% p<0.001). CONCLUSIONS: Persistent impairment of FMD and NMD after repair of coarctation is more likely to be present in patients corrected at older age. It may be an important contributor to abnormal blood pressure regulation and late morbidity and mortality. PMID- 15749192 TI - No relationship between low-density lipoproteins and endothelial function in hemodialysis patients. AB - BACKGROUND: Relationships between low-density lipoprotein cholesterol and endothelial function in hemodialysis patients have yet to be investigated. Furthermore, current reporting of endothelial function data using flow-mediated dilatation has recognised limitations. The aims of the study were to determine the relationship between low-density lipoproteins and endothelial function in hemodialysis patients and to investigate the validity of determining the area under the curve for data collected during the flow-mediated dilatation technique. METHODS: Brachial artery responses to reactive hyperemia (endothelial-dependent) and glyceryl trinitrate (endothelial-independent) were assessed in 19 hemodialysis patients using high-resolution ultrasound. Lipid profiles and other factors known to effect brachial artery reactivity were also measured prior to the flow-mediated dilatation technique. RESULTS: There were no significant relationships between serum low-density lipoproteins and endothelial-dependent or -independent vasodilation using absolute change (mm), relative change (%), time to peak change (s) or area under the curve (mm x s). In hemodialysis patients with atherosclerosis, area under the curve analysis showed a significantly (p<0.05) decreased endothelial-dependent response (mean+/-S.D.: 19.2+/-17.4) compared to non-atherosclerotic patients (42.3+/-28.6). However, when analysing these data using absolute change, relative change or time to peak dilatation, there were no significant differences between the two groups. CONCLUSIONS: In summary, there was no relationship between low-density lipoproteins and endothelial function in hemodialysis patients. In addition, area under the curve analysis of flow-mediated vasodilatation data may be a useful method of determining the temporal vascular response during the procedure. PMID- 15749193 TI - Longitudinal development of psychopathology in an adult congenital heart disease cohort. AB - BACKGROUND: Little is known about the development of psychopathology in patients with congenital heart disease (ConHD) from adolescence into adulthood. In this study, the course of psychopathology in a cohort of ConHD adults, who received their first heart surgery in childhood, was determined longitudinally over a 10 year-period. METHODS: At both the first (1989-1991) and second (2000-2001) follow up, patients (n=251, aged 20-32 years) completed questionnaires during a psychological examination. Parallel instruments were used to measure psychopathology in adolescence and adulthood. RESULTS: Young ConHD female patients showed higher levels of psychopathology compared to the reference group. Larger differences in psychopathology were found between younger (20-27 years) and older (28-32 years) patients compared to the reference group; the younger patients showed more psychopathology than the older patients. Also, larger differences in psychopathology were found between male and female patients compared to the reference group; female patients showed more psychopathology than male patients. Parents and partners of patients evaluated the patients' behavior as more deviant from the norm than patients themselves. There was a stronger decline of psychopathology in ConHD patients with increasing age compared to reference groups. Female and younger patients showed more variability in trajectories of psychopathology than did male and older patients. CONCLUSIONS: Assessment and treatment of psychopathology at a younger age is highly desirable in ConHD patients. Young adult ConHD female patients deserve special attention since they might be hampered by disease-specific uncertainties. PMID- 15749195 TI - Atrioventricular conduction disturbances in a young patient with Fabry's disease without other signs of cardiac involvement. AB - A 30 year-old male patient with a history of Fabry's disease, was referred to hospital with symptoms of dizziness, hypotension and weakness. Fabry's disease had been diagnosed 2 years before, based on angiokeratoma and hypohidrosis on physical examination and complete lack of alpha-galactosidase A on laboratory examination. The ECG on admission demonstrated sinus bradycardia, with a poor response to atropine administration. Echocardiograms on admission and 2 years before were normal, as well as Holter ambulatory ECG recording. Subsequent electrophysiological study demonstrated mild AV conduction disturbances at a site proximal to His, and the patient was simply advised to be regularly followed up. It can therefore be concluded that even young patients with Fabry's disease and normal echocardiograms might develop cardiac symptoms due to AV conduction abnormalities. PMID- 15749197 TI - Ruptured abdominal aortic aneurysm with Marfan's syndrome variant, a case from the family with high incidence of aortic aneurysm. PMID- 15749196 TI - Intracranial aneurysms, coronary aneurysms and descending aortic coarctation- unreported association. AB - Rare association of coronary artery aneurysms with intra cranial aneurysms is reported. Also, association of abdominal aortic coarctation with intracranial aneurysms is rare. A 70-year-old female presented with subarachnoid hemorrhage secondary to rupture of intracranial aneurysm. On evaluation, she was found to have intracranial aneurysms in the vertebral and basilar artery, coronary aneurysms and descending thoracic aortic coarctation. This association is unreported. PMID- 15749194 TI - Rescue percutaneous coronary intervention for failed thrombolysis in a patient with anomalous coronary arteries. AB - Coronary artery anomalies (CAA) often render technically difficult the completion of coronary angiography and intervention. Their presence in patients undergoing emergency angiography for acute myocardial infarction (AMI) is particularly challenging for interventional cardiologists. In this article, we report, for the first time in the literature, a case of rescue percutaneous coronary intervention for failed thrombolysis in a patient with AMI due to occlusion of a left circumflex coronary artery with anomalous origin from right sinus of Valsalva (in an anomalous left coronary system also including an anomalous origin of the left anterior descending artery from the right sinus). In particular, the present clinical vignette emphasizes the importance of a thorough search for the culprit vessel during cardiac catheterization. Especially in the emergency setting, non invasive methods of ischemia localization, such as ST-segment elevation at the ECG and wall motion abnormalities at echocardiography, are of pivotal usefulness to guide the interventional cardiologist in identifying and treating the diseased coronary vessel in a timely and effective fashion. PMID- 15749198 TI - Prolonged QT interval with markedly abnormal ventricular repolarization in diphenhydramine overdose. AB - Diphenhydramine is a histamine-1 receptor antagonist of ethanolamine origin with anticholinergic, sedative, antivertigo, antiemetic, antidyskinetic, and local anesthetic properties. It inhibits the fast sodium channels and, at higher concentrations, also the potassium channels, inhibition of which may result in QT interval prolongation on electrocardiogram. The markedly abnormal ventricular repolarization is rare with diphenhydramine overdose. We report a case where a young woman who took moderate overdose of diphenhydramine (625 mg) along with acetaminophen developed prolonged QT interval with strikingly abnormal T waves. These changes reverted to normal with treatment. Patient did not experience torsade de pointes, possibly secondary to the protective effect of the diphenhydramine overdose-related tachycardia. As the tachycardia caused by diphenhydramine overdose seems to protect against torsade de pointes, it may be practical to avoid bradycardia in acute phase of diphenhydramine toxicity. Acetaminophen has not been shown to prolong QT interval or effect cardiac repolarization. PMID- 15749199 TI - Hypoxia inducible factor-1 expression mediates myocardial response to ischemia late after acute myocardial infarction. AB - We report hypoxia-inducible factor-1 (HIF-1) expression in myocardium of patients with recent acute myocardial infarction (AMI), supporting the hypothesis of HIF-1 as a possible mediator of response to ischemia. A potential diagnostic role of determining tissue expression of HIF-1 as a marker of ischemia, and potential therapeutic implications by trying to modulate HIF-1 activity in order to promote beneficial effects of HIF-1 related genes (e.g. expression of vascular endothelial growth factor (VEGF)) may derive. PMID- 15749200 TI - Suppression by propranolol and amiodarone of an electrical storm refractory to metoprolol and amiodarone. AB - A 60-year-old male patient with ischemic cardiomyopathy experienced an electrical storm 1 month after implantation of an internal cardioverter-defibrillator. Recurrent life-threatening episodes of ventricular tachycardia persisted despite maximal antiarrhythmic theraphy with amiodarone, metoprolol and mexiletine. After a total of more than 500 cardioversions, all antiarrhythmic medications were withdrawn, and the nonselective beta-blocker propranolol was initiated. Electrical stability was achieved and the patient was discharged on propranolol 400 mg/day. Two months later, a second arrhythmia cluster occurred that was controlled by the addition of amiodarone. The patient remains free of arrhythmia 15 months after the event with the combination of propranolol and amiodarone. PMID- 15749201 TI - A novel mutation in FKBP12.6 binding region of the human cardiac ryanodine receptor gene (R2401H) in a Japanese patient with catecholaminergic polymorphic ventricular tachycardia. AB - Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an autosomal dominant inherited disorder characterized by adrenergic induced polymorphic ventricular tachycardias and associated with sudden cardiac death. The human cardiac ryanodine receptor gene (RyR2) was linked to CPVT. A 20-year-old male was referred to our hospital because of recurrent syncope after physical and emotional stress. Routine cardiac examinations including catheterization revealed no structural abnormality. Exercise on treadmill induced premature ventricular contraction in bigeminy and bidirectional ventricular tachycardia was induced during isoproterenol infusion. Beta-blocking drug was effective in suppressing the arrhythmias. We performed genetic screening by PCR-SSCP method followed by DNA sequencing, and a novel missense mutation R2401H in RyR2 located in FKBP12.6 binding region was identified. This mutation was not detected in 190 healthy controls. Since FKBP12.6 plays a critical role in Ca channel gating, the R2401H mutation can be expected to alter Ca-induced Ca release and E-C coupling resulting in CPVT. This is the first report of RyR2 mutation in CPVT patient from Asia including Japan. PMID- 15749203 TI - Coronary rotational ablation for calcific coronary artery stenosis in a young child. AB - Coronary aneurysm formation and subsequent stenosis resulting in ischemic heart disease are the most important complications of Kawasaki disease (KD). Reported is the evolution of severe calcific coronary artery stenosis in an 8-year-old boy managed with percutaneous transluminal coronary rotational ablation (PTCRA) after failed balloon angioplasty. No restenosis occurred 6 months after RA. Reports on PTCRA in children are limited and it may be a good therapeutic option as a definite or a palliative one to postpone coronary artery bypass surgery in the young children with calcific coronary artery stenosis after Kawasaki disease. PMID- 15749202 TI - A rare cause of atrioventricular block: Mad Honey intoxication. PMID- 15749204 TI - Late anthracyclines cardiotoxicity associated with infection by Mycoplasma pneumoniae. A case report. AB - We report the case of a 15-year-old male developing progressive myocardiopathy secondary to late anthracycline cardiotoxicity. At 2 years of age, the patient received chemotherapy with anthracyclines (adriamycin) at an accumulated dose of 510 mg/m2 due to the presence of non-Hodgkin lymphoma which subsided completely and he has been asymptomatic since then. A number of recurrent respiratory infections occurred as triggering factors of the condition, and the laboratory tests evidenced positive serology for Mycoplasma pneumoniae suggesting a recent contact. PMID- 15749205 TI - Acute myocardial infarction with normal coronary arteries in a young man with the Behcet's disease. AB - Behcet's disease (BD) is an inflammatory disorder of unknown origin, which usually presents with mucocutaneous, ocular, articular, vascular, gastrointestinal and central nervous system manifestations. Although cardiac involvement is not infrequent as a manifestation of Behcet's disease, coronary arteritis is very rarely reported. We suggest that the diagnosis of coronary arteritis should be considered in patients presenting acute myocardial infarction especially in young patients as underlying cause. PMID- 15749206 TI - Swyer-James-MacLeod syndrome complicated by spontaneous multivessel coronary dissection. PMID- 15749207 TI - Recurrent coronary spasm with complete atrioventricular block in a patient with refractory acute myelocytic leukemia. PMID- 15749208 TI - Three-dimensional finite element analysis used to compare methods of fixation after sagittal split ramus osteotomy: setback surgery-posterior loading. AB - We used three-dimensional finite element analysis to compare the biomechanical stability of bilateral sagittal split ramus osteotomies fixed by lag screws with linear and triangular configuration, and double or single six-hole miniplates with monocortical screws after set-back operation. The three-dimensional finite element model contained 122,717 elements and 25,048 nodes. Posterior occlusal loads were simulated on the distal segments. MSC Marc software was used to calculate the stress fields on both the segments and the fixing appliances. We conclude that either triangular lag screw configuration or double miniplates led to better stability and lower mechanical stresses near the osteotomy than the linear lag screws or single oblique miniplates. PMID- 15749209 TI - Bilateral mandibular distraction in adult dogs with an epiperiosteal distractor. AB - We tested the function and efficiency of a new intraoral, bone-borne distraction device for mandibular lengthening, and the effect of bilateral transsection of the inferior alveolar arteries on osteogenesis in adult dogs. Nine adult Beagles had bilateral complete mandibular osteotomies and epiperiosteal placement of the distractor. After 7 days, the mandibles were distracted at 1 x 1mm/day, 1 x 0.5 mm/day or 2 x 0.25 mm/day for 15 or 30 days, respectively. Hemimandibles were harvested after 1, 14, or 42 days consolidation and evaluated clinically, radiographically and histologically. The mean distraction achieved was 11 mm (range 4-16 mm). New bone formation and clinical stability increased with increasing consolidation time and was independent of the distraction protocol used. Osteogenesis at the distal and proximal mandibular fragments did not differ. In 10/18 distraction sites, wound dehiscences compromised osteogenesis. The new epiperiosteal distractor permitted successful distraction osteogenesis despite complete bilateral osteotomy with section of the medullary vessels in adult dogs. PMID- 15749210 TI - Aplasia of the parotid gland in Down syndrome. AB - Salivary gland aplasia has not to our knowledge been previously reported in association with Down syndrome. We present a case of bilateral parotid aplasia in a patient with Down syndrome. Clinically he had aplasia of the major salivary glands and symptoms of xerostomia. Thirteen other family members over three generations were examined, and all had functional parotid glands. We reviewed publications about Down syndrome and salivary aplasia, together with the data regarding his other clinical problems and family background. His oral problems were inadequate plaque control, dental caries, and erosion of the teeth. PMID- 15749211 TI - Follow up of mandibular costochondral grafts after release of ankylosis of the temporomandibular joints. AB - The records of 55 patients with ankylosis of the temporomandibular joint (25 unilateral and 30 bilateral); were selected at random. In these patients, the resected condyles had been replaced with costochondral grafts and the patients were followed clinically and radiographically for 7-10 years. Of the 85 grafts, we found take and good remodeling in 50 (59%), reankylosis in 8 (9%), resorption of the graft in 21 (25%) and overgrowth of the graft in 3 (4%). Mouth opening was satisfactory (more than 25 mm) in 32 of the 55 patients (58%), unsatisfactory (between 5 and 25 mm) in 10 (18%), and the operation was a failure in 13 (24%). PMID- 15749212 TI - Delayed tension pneumocephalus complicating orbital exenteration. AB - A 43-year-old woman with a recurrent adenoid cystic carcinoma of the lacrimal gland was treated by orbital exenteration and reconstruction. Eight days later she developed symptoms of cerebral irritation and a computed tomogram showed intracerebral and subarachnoid air. She recovered with conservative treatment. PMID- 15749213 TI - Management of advanced and recurrent squamous cell carcinoma of the inner nasal lining. AB - Ten cases of advanced and eight cases of recurrent carcinomas of the inner nasal lining were managed in our department between June 1996 and June 2003. Six tumours were managed by operation alone, four by a combination of operation and radiotherapy, two by a combination of chemotherapy and operation, and five by a combination of chemotherapy, operation and radiotherapy. One patient died before the end of treatment. One patient had a combination of chemotherapy and radiotherapy. Eleven patients had reconstructions. RESULTS: After a mean follow up of 2.7 years (range: 3 months to 7 years), 15 patients were disease-free. Two patients had died of their disease, and one had died of an unrelated cause. Reconstruction was satisfactory in six of the eight cases that were evaluated. Prostheses were satisfactory in four of six cases. PMID- 15749214 TI - The relation between health-related quality of life, past medical history, and American Society of Anesthesiologists' ASA grade in patients having primary operations for oral and oropharyngeal cancer. AB - Pre-existing medical problems have the potential to affect postoperative survival, complications, and health-related quality of life (QoL). Our aim was to explore the relation between past medical history, American Society of Anesthesiologists' (ASA) score, health-related QoL, and survival. We collected data from 278 consecutive patients with previously untreated oral and oropharyngeal squamous cell carcinoma operated on primarily from 1995 to 1999 inclusive. Past medical history was recorded from the case notes, ASA grade from the anaesthetic record, and QoL was measured using the University of Washington Quality of Life Questionnaire (UW-QoL). Responses to questionnaires were received from (71%) at baseline (63%) at 6 months (73%) at 1 year, and (65%) 18 months or longer. Past medical history was associated with lower ASA scores. At baseline both history and ASA scores were related to the UW-QoL. Longitudinally patients in ASA grade 1 or with no past history scored better in these UW-QoL domains. Past history did not predict survival (P = 0.83), nor did the UW-QoL composite score (P = 0.30), whilst ASA was associated with crude survival (P = 0.003) and disease-specific survival (P = 0.03). When analyses were stratified for adjuvant radiotherapy, type of operation, size of tumour, and age then the relation to ASA was maintained and trends in the past history and UW-QoL remained not significant. ASA, which is often recorded as part of preoperative assessment, reflects both survival and health-related QoL, and is more useful than past history alone for predicting outcome. PMID- 15749215 TI - Role of tonsillectomy in histology for adults with unilateral tonsillar enlargement. AB - Unilateral tonsillar enlargement (UTE) may indicate malignancy. The purpose of this study was to evaluate the necessity of tonsillectomy for histological examination as a routine practice for every such case. We attempted to identify every adult who had tonsillectomy in our unit from January 1991 to January 2000 in whom the indication for operation was unilateral tonsillar enlargement. The case notes and pathology records were reviewed. One hundred and seven cases were identified but 9 case records could not be found, leaving 98 cases for analysis. Forty-eight were men and 50 were women, aged 16-85 (median age 32). Twenty-three malignancies were identified (23%). A high degree of clinical suspicion was noted pre-operatively in 37 cases, which included all 23 malignancies. Malignancy was more common in the presence of ulceration, male sex, lymphadenopathy, and age 45 years or more, and when it was the patient who first noticed the tonsillar enlargement. Current smoking and persistent pain were not significantly associated with malignancy. Night sweats and weight loss were uncommon. In many cases the apparent tonsillar enlargement was spurious due to asymmetry of the tonsillar pillars. PMID- 15749216 TI - Use of the buccal fat pad in the reconstruction and prosthetic rehabilitation of oncological maxillary defects. AB - We evaluated the effectiveness of the buccal fat pad as a pedicled flap for intraoral reconstruction after partial maxillectomy for neoplastic disease in 24 patients, and subsequently, in providing support for a denture. In all patients the buccal fat pad was covered with a split-skin graft and an acrylic plate. There was complete healing of the buccal fat pad flap within 6 weeks in 18 patients with no major complications, and minimal effects on speech and eating. In six cases there was partial dehiscence of the flap, which healed spontaneously in one patient and was repaired with local flaps in two others. There were no cases of complete breakdown of the flap. Eight patients so far have been rehabilitated with small dentures. In conclusion, the buccal fat pad flap is a simple, quick, and reliable method of reconstruction of small to medium-sized posterior maxillary alveolar defects. PMID- 15749217 TI - By what mechanism do leeches help to salvage ischaemic tissues? A review. AB - The therapeutic use of leeches in medicine dates back to 50 b.c. and was cited by ancient authors. The medicinal leech, Hirudo medicinalis, has been used with increasing frequency during the past few years by reconstructive surgeons to help salvage ischaemic tissues. We aim to summarise the anatomy, physiology, and pharmacological mechanisms of action of leeches to provide reconstructive surgeons with a theoretical basis for their use. PMID- 15749218 TI - Repair of radial free flap donor site by full-thickness skin graft from inner arm. AB - Sixty-two consecutive patients had 63 radial free flaps taken and the donor site repaired with either a full-thickness or split-skin graft. A full-thickness graft was harvested from the inner upper arm in 53 patients (85%). The donor site was assessed using a visual analogue scale, tactile sensitivity, and the Vancouver scar assessment score. All donor sites healed without complications except for two minor wound dehiscences. Both patients and surgeons rated the aesthetic outcome as good. The advantages included; single operation site; simple execution; thin hairless graft, good colour match and an inconspicuous scar causing minimal morbidity. PMID- 15749219 TI - Total mandibular reconstruction for massive osteolysis of the mandible (Gorham Stout syndrome). AB - Massive osteolysis (Gorham-Stout syndrome) is a rare condition of unknown aetiology that is thought to result from a localised endothelial proliferation of lymphatic vessels resulting in destruction and absorption of bone. The diagnosis of Gorham-Stout syndrome can be made only after first excluding osteolysis from infection, inflammation, endocrine disease, and cancer. The syndrome is rarely seen in the facial skeleton and has a variable prognosis. Many treatments have been advocated but only surgery and radiotherapy have had some success. We present a case of massive osteolysis of the entire mandible, which was reconstructed with a free vascularised fibular graft and bilateral prosthetic replacements of the temporomandibular (TMJ) joints. Four years later the fibular graft had been absorbed requiring further reconstruction with another fibula graft. PMID- 15749220 TI - Overnight-stay patients in a day surgery unit to overcome shortage of inpatient beds. AB - A dedicated 23-h stay list was organised at the Addenbrooke's Hospital day surgery unit for elective cases in oral and maxillofacial surgery of minor or moderate severity that would normally be done as inpatients. The patients were selected by the two consultant oral and maxillofacial surgeons and the operations were done by a consultant surgeon (L.C.) and a consultant anaesthetist. Over a 14 month trial period 137 patients were admitted to our 23-h stay lists. One hundred and twenty seven of whom were operated on and stayed overnight, nine went home the same day, and one patient stayed beyond the 23-h limit as he was admitted to the medical ward after a grand mal seizure. The operations were for neoplastic, traumatic, salivary gland and jaw lesions. The reasons for using the overnight stay facilities varied from social reasons to the need to monitor the airway. The fact that the theatre slots and overnight beds were guaranteed, and that the admissions were booked at outpatient consultation led to few cancellations by patients. This process reduced the inpatient waiting list and released the main operating theatre for major oral and maxillofacial operation, particularly for cancer. PMID- 15749221 TI - Treatment of chronic sclerosing osteomyelitis of the mandible with calcitonin: a report of two cases. AB - We report two patients with diffuse sclerosing osteomyelitis in the mandible in whom management with non-steroidal anti-inflammatory drugs, long-term antibiotics, and surgical debridement failed to resolve the symptoms of pain and swelling, which subsequently improved when the patients were given calcitonin. PMID- 15749222 TI - Diagnostic difficulties of complex root morphology: case study of a lower wisdom tooth. AB - We did a panoramic radiographic examination of a symptomatic lower third molar for surgical planning and risk assessment. This showed a close association with the inferior dental canal, in addition to uncertain root morphology. A periapical radiograph showed evidence of additional smaller curved roots. Axial computed tomography (CT) with coronal reformats did not provide further information about the morphology of the root. After removal of the tooth, its root form was compared with the diagnostic images. The periapical radiograph provided the most information about the morphology of the additional roots. The axial and reformatted coronal CT showed accurately the position and tortuous course of the inferior dental canal. PMID- 15749223 TI - Migrating salivary stones: report of three cases. AB - Patients with salivary calculi are normally managed by removal of the calculus or, if necessary, the affected gland. If it is left untreated, a stone may migrate into the adjacent tissues. We present three patients in whom salivary calculi tracked to the surface of the skin. Two were removed under local anaesthetic, and the third patient was lost to follow up. PMID- 15749224 TI - Stabilisation of the unstable fractured zygomatic arch with a Kirschner wire. AB - Although relatively uncommon, isolated fractures of the zygomatic arch can sometimes be difficult to stabilise following reduction. We present a simple method of stabilisation using a Kirschner wire. PMID- 15749225 TI - Submucoal lymphoid aggregates of the lower lip in a 10-year-old boy. AB - We report an unusual case of lymphoid hyperplasia that looked like a mucocele in the lower lip of a 10-year-old boy. PMID- 15749226 TI - A simple way to assist placement of a bone graft into a soft tissue tunnel. AB - We describe a simple and cost effective technique for the insertion of calverial bone grafts for augmentation rhinoplasty. PMID- 15749227 TI - Re: Gibbons AJ, Hope DA, Silvester KC, Oral endotracheal intubation in the management of midfacial fractures (Br J Oral Maxillofac Surg 2003;41:259-60). PMID- 15749228 TI - Molecular and genetic aspects of gastrointestinal and pancreatic disease. PMID- 15749229 TI - Early events in acute pancreatitis. AB - Considerable progress in the understanding of the pathogenesis of acute pancreatitis is based on the conclusive finding that the initiation of the disease occurs within the acinar cell. Two lines of evidence have contributed to the progress in understanding the disease process: (1) the identification of patients with a hereditary form of pancreatitis as carriers of germline-mutations in the genes for cationic trypsinogen and the pancreatic secretory trypsin inhibitor and (2) the use of various transgenic and knock-out mouse strains in experimental models of acute pancreatitis. On the other hand, these studies have delivered several unexpected results that appear to be incompatible with long standing dogmas and paradigms of pancreatic research. Further progress in knowledge will result if the well-characterized enzymatic properties of human enzymes that are involved in the initial activation cascade can be investigated under in vivo conditions in transgenic animals or in permanent acinar cell lines. Such studies will permit the development of effective strategies for the prevention and treatment of this disease. PMID- 15749230 TI - Inflammatory markers of disease severity in acute pancreatitis. AB - To date, CRP remains the single standard biochemical marker for predicting the severity of AP. Because the combination of clinico-physiological scores and CRP provides good information at 48 hours, research has focused on the predictive ability of various markers when applied in the initial 24 hours after admission to the hospital. After detailed review of the literature, the authors conclude that there is no single tool that serves as the optimal predictor of severity. There are, however, data supporting the use of certain tests to improve upon the clinician's early predictive ability on the subsequent course of AP. These include an APACHE II score greater than seven, IL-6 at the time of admission, and urine TAP, urine trypsinogen-2,and serum PMN-elastase at 24 hours (Box 1). These markers will only be able to help the clinician's predictive ability if they can be performed locally and if the results can be available in a timely manner. Future research should focus on markers such as procalcitonin, IL-8, IL-I ra, sTNFR,CAPAP, PLA-2, novel markers, and the combined use of more than one marker. The conventional research approach in predicting severity used in the last15 years has limitations and appears to have reached its maximal potential. Novel conceptions and approaches, such as identification of genetic polymorphisms that predispose to severe course and complications of AP, are needed for a quantum step forward. PMID- 15749231 TI - Cationic trypsinogen mutations and pancreatitis. AB - The discovery of PRSS 1 mutations in hereditary pancreatitis and analysis of how the genotype affects the presentation and progression of hereditary pancreatitis has led to a better understanding of the pathophysiology of the disease. Patients with hereditary pancreatitis present with symptoms at an early age and have a significant lifetime risk for the development of endocrine and exocrine insufficiency, albeit at a later stage than patients with either idiopathic or alcoholic chronic pancreatitis. There are distinct phenotypic differences between hereditary pancreatitis and with other types of pancreatitis. As many as 80% of patients with symptomatic hereditary pancreatitis have an underlying causative PRSS1 mutation; there are, however, few significant phenotypic differences between these PRSS1 mutations. TheR122H mutation is the most common PRSS1 mutation observed, and patients with the R122H mutation present earlier. This, however, does not necessarily translate into a more aggressive disease with respect to complications of chronic pancreatitis. Indeed, the age of presentation of symptoms may be a poor surrogate for predicting outcome, as inherited disorders of trypsinogen may cause subclinical attacks of pancreatitis, which ultimately lead to pancreatic destruction and dysfunction. All patients, irrespective of whether they carry a PRSS1 mutation, are at significant risk of developing pancreatic ductal adenocarcinoma. The risk appears to be insignificant below the age of 40 years, but it increases incrementally thereafter. Significantly, the risk of pancreatic cancer is not related to PRSS1 mutation type and does not appear to be related to the mode of inheritance. The role of SPINK1 mutations in modifying the expression of PRSS1mutations is unclear but appears to be of clinical importance. It is unlikely that they act as causative mutations per se, at least in the Western form of the disease. Additionally, they do not appear to have an impact on the penetrance of PRSS1 gene mutations in hereditary pancreatitis. PMID- 15749232 TI - Serine protease inhibitor kazal type 1 mutations and pancreatitis. AB - SPINKI is thought to play an important role in protecting the pancreas against excessive trypsinogen activation. SPINK1 mutations are associated with the development of acute and chronic pancreatitis and have been detected in all forms of chronic pancreatitis. The strong association of mutations in the PRSS1 gene and in the SPINKI gene with chronic pancreatitis supports the concept of intracellular trypsin activation as an initiating and extremely important step in the development of pancreatitis. The N34S mutation represents the most frequently observed pancreatitis-associated SPINKI variant. Because the SPINKI N34Smutation is very common in the general population, it is unlikely that this mutation alone can initiate the development of chronic pancreatitis. Thus, it rather appears that in most patients with SPINKI-associated chronic pancreatitis, this genetic variant acts as disease modifier or within a polygenic model with other yet unidentified genes or environmental co-factors. The possible interaction of mutations in the SPINK1 gene with other pancreatitis-associated susceptibility genes has to be investigated in future research efforts. The most promising candidate gene for such an interaction is the CFTR gene, because genetic alterations within the CFTRgene are also common in the general population and already have been associated with chronic pancreatitis. PMID- 15749233 TI - The impact of cystic fibrosis and PSTI/SPINK1 gene mutations on susceptibility to chronic pancreatitis. AB - This article reviews current concepts regarding the pathobiology of cystic fibrosis pancreatic disease. It summarizes recent studies on the relationship between CFTR mutations and pancreatitis, and it reviews several unresolved issues in the field. PMID- 15749234 TI - Molecular diagnosis of solid and cystic lesions of the pancreas. AB - Pancreatic cancer presents at a clinically advanced stage. Diagnosis often is delayed in those who undergo several negative investigations, and at times, the diagnosis ultimately is made at surgery. Tumor suppressor-linked microsatellite allelic loss analysis appears to improve the diagnostic accuracy of inconclusive cytology specimens. Science, however, still suffers from the lack of a reliable biomarker for the purpose of screening and early detection. The continued study of sera and pancreatic juice hopefully will lead to the discovery of molecular biomarkers capable of diagnosing preclinical disease in at-risk groups.The ability to predict the underlying biologic behavior of pancreatic cystic neoplasia is less than ideal. Tumor suppressor-linked microsatellite allelic loss and telomerase analysis show promise, but they have not been studied extensively. Proteomic analysis of pancreatic cyst fluid, although not yet reported, also may prove valuable in guiding clinical management. PMID- 15749235 TI - Genetics of hereditary pancreatic carcinoma. AB - Several tumor syndromes are known to have an increased incidence of pancreatic carcinoma. Unfortunately, the incidence of pancreatic carcinoma almost equals its mortality. Because of intense research activities in the past decade, several genetic alterations have been identified that are responsible for the so-called sporadic pancreatic carcinoma development. National and international research activities only recently have begun focusing on the molecular pathogenesis of hereditary risk factors for pancreatic carcinoma. It is hoped that these genetic studies will aid the development of clinical management strategies for pancreatic carcinoma prevention. PMID- 15749236 TI - From colonic polyps to colon cancer: pathophysiology, clinical presentation, and diagnosis. AB - Epidemiologists, basic researchers, clinicians, and public health administrators unite! Develop and implement a simple, safe, and effective preventive and screening test for colon cancer. The public will willingly and enthusiastically accept such a test. Many thousands of lives are at stake every year. PMID- 15749237 TI - Microsatellite instability and DNA mismatch repair deficiency testing in hereditary and sporadic gastrointestinal cancers. AB - The reference cancers associated with DNA mismatch repair (MMR)deficiency are the adenocarcinomas of patients with hereditary nonpolyposis colorectal cancer, also known as Lynch syndrome. Sporadic gastrointestinal (GI) carcinomas, most commonly colorectal and gastric carcinomas, may also be associated with deficiencies of DNA mismatch repair. Deficiency in cellular MMR leads to wide-spread mutagenesis and neoplastic development and progression. An important diagnostic feature of MMR-deficient tumors is the high rate of mutations that accumulate in repetitive nucleotide regions, and these mutations are known as microsatellite instability(MSI). A standard panel of markers to test for MSI in tumors has been recommended and efficiently separates tumors into those with high, low, or no microsatellite instability (MSI-H, MSI-L, or MSS). Tumors characterized by MSI-H characteristically show loss of one of the main DNA MMR proteins, mLH1 or MSH2, and rarely MSH6 and PMS2, detected by immunohistochemistry (IHC). The combination of MSI testing and IHC for MMR proteins in tumors tissues is used to identify underlying DNA MMR deficiency andis clinically relevant screen patients who might have hereditary non-polyposis colorectal cancer for DNA repair gene germline testing. Increasing evidence demonstrates that tumors with a positive MSI status have lower lymph node metastases burden, and these patients have an overall improved survival, suggesting that the MSI and MMR status may contribute to decision making regarding treatment approaches. Updated guidelines for MSI and IHC for DNAMMR testing, and the biological and potential clinical implications of MMR deficiency and microsatellite instability in GI polyps and cancers are reviewed. PMID- 15749238 TI - Molecular markers in Helicobacter pylori-associated gastric carcinogenesis. AB - Helicobacter pylori infection is a known risk factor of gastric carcino-genesis. This article presents early molecular alterations associated with H. pylori chronic gastritis and advances in the molecular characterization of preneoplastic intestinal metaplasia (IM) and premalignant gastric mucosal lesions. H. pylori infection induces changes in gene expression, genomic instability and accumulation of gene mutations in the stomach epithelium. Mutations, including LOH and microsatellite instability, and gene hypermethylation are seen not only in gastric cancer, but are already detectable in IM and gastric dysplasia/adenoma. Recent reports using microarray expression analysis identified several gastric epithelial genes that are regulated by H. pylori. Among the many genes showing altered epithelial expression in response to H. pylori, some might be useful as markers to assess gastric cancer risk. Profiles of mutagenesis and gene expression in IM and dysplasia/adenoma have been characterized and represent potential markers of preneoplastic and premalignant lesions during gastric carcinogenesis. PMID- 15749240 TI - Developmental deficits in social perception in autism: the role of the amygdala and fusiform face area. AB - Autism is a severe developmental disorder marked by a triad of deficits, including impairments in reciprocal social interaction, delays in early language and communication, and the presence of restrictive, repetitive and stereotyped behaviors. In this review, it is argued that the search for the neurobiological bases of the autism spectrum disorders should focus on the social deficits, as they alone are specific to autism and they are likely to be most informative with respect to modeling the pathophysiology of the disorder. Many recent studies have documented the difficulties persons with an autism spectrum disorder have accurately perceiving facial identity and facial expressions. This behavioral literature on face perception abnormalities in autism is reviewed and integrated with the functional magnetic resonance imaging (fMRI) literature in this area, and a heuristic model of the pathophysiology of autism is presented. This model posits an early developmental failure in autism involving the amygdala, with a cascading influence on the development of cortical areas that mediate social perception in the visual domain, specifically the fusiform "face area" of the ventral temporal lobe. Moreover, there are now some provocative data to suggest that visual perceptual areas of the ventral temporal pathway are also involved in important ways in representations of the semantic attributes of people, social knowledge and social cognition. Social perception and social cognition are postulated as normally linked during development such that growth in social perceptual skills during childhood provides important scaffolding for social skill development. It is argued that the development of face perception and social cognitive skills are supported by the amygdala-fusiform system, and that deficits in this network are instrumental in causing autism. PMID- 15749241 TI - Behavioral manifestations of autism in the first year of life. AB - In the interest of more systematically documenting the early signs of autism, and of testing specific hypotheses regarding their underlying neurodevelopmental substrates, we have initiated a longitudinal study of high-risk infants, all of whom have an older sibling diagnosed with an autistic spectrum disorder. Our sample currently includes 150 infant siblings, including 65 who have been followed to age 24 months, who are the focus of this paper. We have also followed a comparison group of low-risk infants. Our measures include a novel observational scale (the first, to our knowledge, that is designed to assess autism-specific behavior in infants), a computerized visual orienting task, and standardized measures of temperament, cognitive and language development. Our preliminary results indicate that by 12 months of age, siblings who are later diagnosed with autism may be distinguished from other siblings and low-risk controls on the basis of: (1) several specific behavioral markers, including atypicalities in eye contact, visual tracking, disengagement of visual attention, orienting to name, imitation, social smiling, reactivity, social interest and affect, and sensory-oriented behaviors; (2) prolonged latency to disengage visual attention; (3) a characteristic pattern of early temperament, with marked passivity and decreased activity level at 6 months, followed by extreme distress reactions, a tendency to fixate on particular objects in the environment, and decreased expression of positive affect by 12 months; and (4) delayed expressive and receptive language. We discuss these findings in the context of various neural networks thought to underlie neurodevelopmental abnormalities in autism, including poor visual orienting. Over time, as we are able to prospectively study larger numbers and to examine interrelationships among both early-developing behaviors and biological indices of interest, we hope this work will advance current understanding of the neurodevelopmental origins of autism. PMID- 15749242 TI - Brain overgrowth in autism during a critical time in development: implications for frontal pyramidal neuron and interneuron development and connectivity. AB - While abnormalities in head circumference in autism have been observed for decades, it is only recently that scientists have begun to focus in on the developmental origins of such a phenomenon. In this article we review past and present literature on abnormalities in head circumference, as well as recent developmental MRI studies of brain growth in this disorder. We hypothesize that brain growth abnormalities are greatest in frontal lobes, particularly affecting large neurons such as pyramidal cells, and speculate how this abnormality might affect neurofunctional circuitry in autism. The relationship to clinical characteristics and other disorders of macrencephaly are discussed. PMID- 15749244 TI - Neuroanatomic observations of the brain in autism: a review and future directions. AB - Infantile autism is a behaviorally defined disorder associated with characteristic cognitive, language and behavioral features. Several postmortem studies have highlighted areas of anatomic abnormality in the autistic brain. Consistent findings have been observed in the limbic system, cerebellum and related inferior olive. In the limbic system, the hippocampus, amygdala and entorhinal cortex have shown small cell size and increased cell packing density at all ages, suggesting a pattern consistent with development curtailment. Findings in the cerebellum have included significantly reduced numbers of Purkinje cells, primarily in the posterior inferior regions of the hemispheres. A different pattern of change has been noted in the vertical limb of the diagonal band of broca, cerebellar nuclei and inferior olive with plentiful and abnormally enlarged neurons in the brains of young autistic subjects, and in adult autistic brains, small, pale neurons that are reduced in number. These findings combined with reported age-related changes in brain weight and volume, have raised the possibility that the neuropathology of autism may represent an on-going process. PMID- 15749243 TI - Significance of abnormalities in developmental trajectory and asymmetry of cortical serotonin synthesis in autism. AB - The role of serotonin in prenatal and postnatal brain development is well documented in the animal literature. In earlier studies using positron emission tomography (PET) with the tracer alpha[(11)C]methyl-l-tryptophan (AMT), we reported global and focal abnormalities of serotonin synthesis in children with autism. In the present study, we measured brain serotonin synthesis in a large group of autistic children (n = 117) with AMT PET and related these neuroimaging data to handedness and language function. Cortical AMT uptake abnormalities were objectively derived from small homotopic cortical regions using a predefined cutoff asymmetry threshold (>2 S.D. of normal asymmetry). Autistic children demonstrated several patterns of abnormal cortical involvement, including right cortical, left cortical, and absence of abnormal asymmetry. Global brain values for serotonin synthesis capacity (unidirectional uptake rate constant, K-complex) values were plotted as a function of age. K-complex values of autistic children with asymmetry or no asymmetry in cortical AMT uptake followed different developmental patterns, compared to that of a control group of non-autistic children. The autism groups, defined by presence or absence and side of cortical asymmetry, differed on a measure of language as well as handedness. Autistic children with left cortical AMT decreases showed a higher prevalence of severe language impairment, whereas those with right cortical decreases showed a higher prevalence of left and mixed handedness. Global as well as focal abnormally asymmetric development in the serotonergic system could lead to miswiring of the neural circuits specifying hemispheric specialization. PMID- 15749245 TI - The teratology of autism. AB - Autism spectrum disorders affect behaviors that emerge at ages when typically developing children become increasingly social and communicative, but many lines of evidence suggest that the underlying alterations in the brain occur long before the period when symptoms become obvious. Studies of the behavior of children in the first year of life demonstrate that symptoms are often detectable in the first 6 months. The environmental factors known to increase the risk of autism have critical periods of action during embryogenesis. Minor malformations that occur frequently in people with autism are known to arise in the same stages of development. Anomalies reported from histological studies of the brain are consistent with an early alteration of development. Congenital syndromes with high rates of autism include somatic that originate early in the first trimester. In addition, it is possible to duplicate a number of anatomic and behavioral features characteristic of human cases by exposing rat embryos to a teratogenic dose of valproic acid at the time of neural tube closure. PMID- 15749246 TI - Autism associated with conditions characterized by developmental errors in early embryogenesis: a mini review. AB - Autism is a complex developmental disorder without an established single etiology but with significant contributions from genetic studies, functional research, and neuropsychiatric and neuroradiologic investigations. The purpose of this paper is to review the findings in five studies involving individuals manifesting the characteristic findings of autism spectrum disorder associated with malformations and dysfunctions known to result from early embryogenic defects. These investigations include two associated with teratogens (thalidomide embryopathy, Mobius sequence with misoprostol) and three (most Mobius sequence cases, CHARGE association, Goldenhar syndrome) with no known etiology. These studies suggest that early embryonic development errors often involving cranial nerve palsies, internal and external ear malformations, ophthalmologic anomalies, and a variety of systemic malformations may be associated with autism spectrum disorders statistically more frequently than expected in a normal population. Although the exact time of developmental insult for each condition cannot be identified, the evidence is that it may occur as early as week 4 to 6+ of embryogenesis. PMID- 15749247 TI - Three autism candidate genes: a synthesis of human genetic analysis with other disciplines. AB - Autism is a particularly complex disorder when considered from virtually any methodological framework, including the perspective of human genetics. We first present a review of the genetic analysis principles relevant for discussing autism genetics research. From this body of work we highlight results from three candidate genes, REELIN (RELN), SEROTONIN TRANSPORTER (5HTT), and ENGRAILED 2 (EN2) and discuss the relevant neuroscience, molecular genetics, and statistical results that suggest involvement of these genes in autism susceptibility. As will be shown, the statistical results from genetic analysis, when considered alone, are in apparent conflict across research groups. We use these three candidate genes to illustrate different problems in synthesizing results from non overlapping research groups examining the same problem. However, when basic genetic principles and results from other scientific disciplines are incorporated into a unified theoretical framework, at least some of the difficulties with interpreting results can be understood and potentially overcome as more data becomes available to the field of autism research. Integrating results from several scientific frameworks provides new hypotheses and alternative data collection strategies for future work. PMID- 15749248 TI - Neuropeptides and the social brain: potential rodent models of autism. AB - Conducting basic scientific research on a complex psychiatric disorder, such as autism, is a challenging prospect. It is difficult to dissociate the fundamental neurological and psychological processes that are disturbed in autism and, therefore, it is a challenge to discover accurate and reliable animal models of the disease. Because of their role in animal models of social processing and social bonding, the neuropeptides oxytocin and vasopressin are strong candidates for dysregulation in autism. In this review, we discuss the current animal models which have investigated oxytocin and vasopressin systems in the brain and their effects on social behavior. For example, mice lacking the oxytocin gene have profound deficits in social processing and social recognition, as do rats lacking vasopressin or mice lacking the vasopressin V1a receptor (V1aR). In another rodent model, monogamous prairie voles are highly social and form strong pair bonds with their mates. Pair bonds can be facilitated or disrupted by perturbing the oxytocin and vasopressin systems. Non-monogamous vole species that do not pair bond have different oxytocin and V1aR distribution patterns in the brain than monogamous vole species. Potential ties from these rodent models to the human autistic condition are then discussed. Given the hallmark disturbances in social function, the study of animal models of social behavior may provide novel therapeutic targets for the treatment of autism. PMID- 15749249 TI - Neuropeptides and non-human primate social deficits associated with pathogenic rearing experience. AB - There is a persuasive evidence that autism is highly heritable and likely to be substantially determined by polygenic mechanisms. Nevertheless, some intriguing findings in children raised in conditions of extreme social deprivation suggest that an autistic-like syndrome may occur as a consequence of environmental conditions. A particularly close model of this human syndrome has been studied in rhesus monkeys for almost half a century. Monkeys reared in pathogenic rearing conditions manifest considerable deficits in social interaction and increased self-directed behaviors. We have been interested in the possibility that disruptions in normal social development in non-human primates might be expressed in neuropeptide systems which have emerged in rodent studies as important candidates for a unique social biology. In recent studies, we have described persistently reduced CSF OT levels in male rhesus monkeys with significant social deficits. We also found that OT levels were positively related to the expression of affiliative social behaviors. Alterations were also detected in both CRH and AVP receptor binding patterns in limbic structures likely to influence social and emotional development. Taken together, these data suggest that abnormal rearing influences the development of brain systems critical to normal social and emotional competence in rhesus monkeys and may contribute to the development of autistic-like symptomatology associated with pathogenic rearing histories. PMID- 15749250 TI - Is autism due to brain desynchronization? AB - The hypothesis is presented that a disruption in brain synchronization contributes to autism by destroying the coherence of brain rhythms and slowing overall cognitive processing speed. Particular focus is on the inferior olive, a precerebellar structure that is reliably disrupted in autism and which normally generates a coherent 5-13 Hz rhythmic output. New electrophysiological data reveal that the continuity of the rhythmical oscillation in membrane potential generated by inferior olive neurons requires the formation of neuronal assemblies by the connexin36 protein that mediates electrical synapses and promotes neuronal synchrony. An experiment with classical eyeblink conditioning is presented to demonstrate that the inferior olive is necessary to learn about sequences of stimuli presented at intervals in the range of 250-500 ms, but not at 700 ms, revealing that a disruption of the inferior olive slows stimulus processing speed on the time scale that is lost in autistic children. A model is presented in which the voltage oscillation generated by populations of electrically synchronized inferior olivary neurons permits the utilization of sequences of stimuli given at, or faster than, 2 per second. It is expected that the disturbance in inferior olive structure in autism disrupts the ability of inferior olive neurons to become electrically synchronized and to generate coherent rhythmic output, thereby impairing the ability to use rapid sequences of cues for the development of normal language skill. Future directions to test the hypothesis are presented. PMID- 15749251 TI - A question of balance: a proposal for new mouse models of autism. AB - Autism spectrum disorder (ASD) represents a major mental health problem with estimates of prevalence ranging from 1/500 to 1/2000. While generally recognized as developmental in origin, little to nothing is certain about its etiology. Currently, diagnosis is made on the basis of a variety of early developmental delays and/or regressions in behavior. There are no universally agreed upon changes in brain structure or cell composition. No biomarkers of any type are available to aid or confirm the clinical diagnosis. In addition, while estimates of the heritability of the condition range from 60 to 90%, as of this writing no disease gene has been unequivocally identified. The prevalence of autism is three to four-fold higher in males than in females, but the reason for this sexual dimorphism is unknown. In light of all of these ambiguities, a proposal to discuss potential animal models may seem the heart of madness. However, parsing autism into its individual genetic, behavioral, and neurobiological components has already facilitated a 'conversation' between the human disease and the neuropathology and biochemistry underlying the disorder. Building on these results, it should be possible to not just replicate one aspect of autism but to connect the developmental abnormalities underlying the ultimate behavioral phenotype. A reciprocal conversation such as this, wherein the human disease informs on how to make a better animal model and the animal model teaches of the biology causal to autism, would be highly beneficial. PMID- 15749252 TI - Making and breaking serotonin neurons and autism. AB - Dysfunction of brain serotonin system development is hypothesized to contribute to autistic behaviors. The testing of this hypothesis will likely depend on a better understanding of the genes and mechanisms involved in serotonin neuron cell fate specification. In this review we summarize the main features of vertebrate serotonin neuroanatomical development and recent studies that have revealed critical steps in the molecular genetic program that controls serotonin neuron phenotype. We then discuss the potential relevance of these findings to advances in autism research and to new molecular genetic tools under development that will impact future testing of the hypothesis. PMID- 15749253 TI - Maternal administration of thalidomide or valproic acid causes abnormal serotonergic neurons in the offspring: implication for pathogenesis of autism. AB - Embryonic exposure to thalidomide (THAL) or valproic acid (VPA) before neural tube closure has been demonstrated as a useful model for human autism in rats. Abnormalities of the serotonergic system which are often observed in human autism have been shown in these rats. Thus, we examined whether early serotonergic neuronal development is perturbed by THAL/VPA. When pregnant rats were exposed to THAL or VPA on embryonic day 9, a dramatic shift of the distribution of serotonergic neurons in the dorsal raphe nucleus was observed on postnatal day 50. This alteration is thought to reflect abnormality of serotonergic neuronal differentiation and migration. In vitro studies revealed that VPA retards the maturation of serotonergic neuron from ES cell-derived neuronal progenitors, whereas exogenously added Sonic hedgehog, a morphogen that has been implicated in serotonergic cell fate, partially prevented this retardation. These results indicate that disruption of early serotonergic neuronal development might be involved in the etiology of autism. PMID- 15749254 TI - Maternal influenza infection is likely to alter fetal brain development indirectly: the virus is not detected in the fetus. AB - Epidemiological studies have shown that maternal infection can increase the risk for mental illness in the offspring. In a mouse model of maternal respiratory infection with influenza virus, the adult offspring display striking behavioral, pharmacological and histological abnormalities. Although influenza primarily infects the respiratory system, there are reports of viral mRNA and protein in the fetus of infected pregnant animals. To determine the extent of viral spread following maternal respiratory infection, we used RT-PCR to assay various maternal and fetal tissues for influenza A mRNAs coding for neuraminidase, non structural protein 2, nuclear protein and matrix protein. While infected maternal lungs exhibit uniformly very strong signals, placentae are only rarely positive, and viral RNAs are not detectable in fetal brains from infected mothers. Thus, the effects of maternal infection on fetal brain development are likely to be indirect, probably involving the maternal inflammatory response. PMID- 15749256 TI - An allegation of scientific misconduct in the Bucci et al. article concerning the effects of DIMP on mink. PMID- 15749258 TI - Pregnancy outcome after exposure to ranitidine and other H2-blockers. A collaborative study of the European Network of Teratology Information Services. AB - BACKGROUND: Published data on pregnancy outcome after exposure to H2-blockers is scarce. The aim of the present study was to evaluate the data collected by the memberships of the European Network of Teratology Information Services (ENTIS). METHODS: The patients were pregnant women who or whose doctor or midwife did contact a Teratology Information Service for risk assessment after the use of a H2-blocker in pregnancy. The data were prospectively collected, i.e. before the outcome of pregnancy was known. Standardized procedures for data collection were used by each centre. The data of the exposed women were compared to those of a control group exposed to non-teratogenic substances. RESULTS: Data on the outcome of 553 pregnancies with exposure to an H2-blocker were evaluated (ranitidine n=335; cimetidine n=113, famotidine n=75; nizatidine n=15, roxatidine n=15). Most of them had been exposed at least in the first trimester. The incidence of premature deliveries was higher in the exposed group compared to the control group. There was no increase in the incidence of major malformations. Two pregnancies with maternal use of famotidine in early pregnancy were terminated after the prenatal diagnosis of a neural tube defect. CONCLUSION: There is no indication for an increased risk of major malformations after the use of H2 blockers during pregnancy. PMID- 15749259 TI - Non-invasive method to assess genotoxicity of nocodazole interfering with spindle formation in mammalian oocytes. AB - Trisomies due to nondisjunction in oogenesis are still a major cause of genetic diseases in humans. In this study, we analysed spindle morphology of in vitro matured nocodazole-exposed mouse oocytes by novel non-invasive Polscope microscopy, and compared images to those obtained by anti-tubulin immunofluorescence of fixed oocytes. Polscope revealed a reduction in the numbers of oocytes expressing a birefringent spindle, and alterations in spindle morphology at concentrations of nocodazole below those inducing detectable aberrations in immunofluorescence. Hyperploidy increased significantly at a concentration of 40 nM nocodazole in mouse metaphase II oocytes, similar to thresholds inducing nondisjunction in cultured human lymphocytes. In conclusion, Polscope represents a novel highly sensitive, non-invasive method to identify chemicals inducing severe spindle aberrations that predispose mammalian oocytes to nondisjunction. Polscope may provide information on the functionality of the spindle in experimental studies but is also compatible with clinical trials in human assisted reproduction due to its non-invasive nature. PMID- 15749260 TI - Effect of nicotine on Oct-4 and Rex-1 expression of mouse embryonic stem cells. AB - To examine the influence that nicotine exposure has on early embryo development, the present study has applied real-time RT-PCR to investigate changes in Oct-4 and Rex-1 expression in mouse embryonic stem (ES) cells exposed to nicotine alone or in the presence of tubocurarine. Oct-4 is regarded as a candidate master regulator gene for the initiation, maintenance, and differentiation of pluripotent cells. Zfp42/Rex-1, another specific gene of pluripotent cells, also plays a critical role in maintaining stem cell character and pluripotency. Results indicated that nicotine (10-1000 nM) enhanced Oct-4 and Rex-1 expression without altering beta-actin expression. This up-regulation with nicotine (100 1000 nM) was prevented with tubocurarine (10 microM), a nicotinic acetylcholine receptor (nAChRs) antagonist. We conclude that nicotine may influence Oct-4 and Rex-1 expression in ES cells through a mechanism involving the nAChRs. PMID- 15749261 TI - Cadmium-induced postaxial forelimb ectrodactyly: association with altered sonic hedgehog signaling. AB - Administration of CdSO(4) to C57BL/6 mice at day 9.5 of gestation induces a high incidence of postaxial forelimb ectrodactyly in the offspring. We propose that Cd(2+) exposure impairs the process of anterior/posterior formation in the limb bud, a process that is directed by Sonic hedgehog (Shh) signaling. We show that exposure of the mouse embryo to Cd(2+) disrupts Shh signaling as measured by polarizing activity of mouse limb bud ZPA grafted to a host chick wing, and activity of a Gli:luciferase reporter exposed to limb bud lysates. Yet the expression of Shh and its translation are not affected by Cd(2+) exposure. We propose that teratogen exposure affects the processing of Shh in the cells in which it is made. PMID- 15749262 TI - Oxidative stress in pregnant women and birth weight reduction. AB - The purpose of this study is to evaluate the role of maternal oxidative stress in lowering neonatal birth weight. Women (N=261) with singleton pregnancy were analyzed for biomarker levels of oxidative stress after recruitment at the time of hospitalization for delivery in Korea between 2000 and 2001. Among the neonates, 247 births were full-term infants and 14 births were pre-term infants. Biomarkers measured for oxidative stress were maternal urinary 8 hydroxydeoxyguanosine (8-OH-dG) and malondialdehyde (MDA). The women with pre term infants had higher concentrations of urinary 8-OH-dG and MDA than those with full-term babies. The concentrations of maternal urinary 8-OH-dG and MDA were inversely associated with birth weight of full-term deliveries after adjusting for potential confounders including maternal age, body mass index, dietary intake, alcohol consumption, smoking exposure, occupational status, and neonatal sex (P<0.05). This study demonstrates that increase of 8-OH-dG and MDA concentrations in urine of pregnant women were associated with reduced birth weight in full-term deliveries. PMID- 15749263 TI - Secondary preeclampsia in rats with nephrotic syndrome -- experimental model. AB - The purpose of the study was to investigate the influence that pregnancy has on the progression of primary nephrotic syndrome (NS). We investigated in pregnant rats the clinical characteristics of NS, including urinary protein concentration, serum albumin, cholesterol, triglycerides, urea, creatinine and electrolytes (potassium, sodium, calcium). Proteinuria and edema increasing during pregnancy, hyponatremia, and also exudates to body cavities were signs of secondary preeclampsia. Hypertension appeared in the second half of pregnancy (from the 10th day) and increased during the perinatal period. All signs of preeclampsia disappeared after puerperium, leaving only the signs of NS existing before pregnancy. Primary, idiopathic gestosis occurs only in humans; therefore, research on that type of gestosis should be confirmed clinically. PMID- 15749264 TI - Effect of the herbicide glyphosate on liver lipoperoxidation in pregnant rats and their fetuses. AB - Glyphosate is a post-emergence herbicide that acts on the synthesis of amino acids and other endogenous metabolites in plants. It is commonly used in agriculture, forestry, and nurseries for the control or destruction of herbaceous plants. Metabolic processes during development and pregnancy could be sensitive to changes induced by glyphosate such as lipid peroxidation. The present study has investigated the effects that 1% glyphosate oral exposure has on lipoperoxidation and antioxidant enzyme systems in the maternal serum and liver of pregnant rats and their term fetuses at 21 days of gestation. The results suggest that excessive lipid peroxidation induced with glyphosate ingestion leads to an overload of maternal and fetal antioxidant defense systems. PMID- 15749265 TI - Antiandrogenic effects in male rats perinatally exposed to a mixture of di(2 ethylhexyl) phthalate and di(2-ethylhexyl) adipate. AB - Di(2-ethylhexyl) phthalate (DEHP) is a well-known testicular toxicant inducing adverse effects in androgen responsive tissues. Therefore, di(2-ethylhexyl) adipate (DEHA) is currently being evaluated as a potential substitute for DEHP. Similarities in structure and metabolism of DEHP and DEHA have led to the hypothesis that DEHA can modulate the effects of DEHP. Wistar rats were gavaged with either vehicle, DEHP (300 or 750mg/kg bw/day) or DEHP (750mg/kg bw/day) in combination with DEHA (400mg/kg bw/day) from gestation day (GD) 7 to postnatal day (PND) 17. Decreased anogenital distance (AGD) and retention of nipples in male offspring were found in all three exposed groups. Dosed males exhibited decreased weights of ventral prostate and m. levator ani/bulbocavernosus. Histopathological investigations revealed alterations in testis morphology in both juvenile and adult animals. The litter size was decreased and postnatal mortality was increased in the combination group only, which is likely a combined effect of DEHP and DEHA. However, no combination effect was seen with respect to antiandrogenic effects, as males receiving DEHP in combination with DEHA did not exhibit more pronounced effects in the reproductive system than males receiving DEHP alone. PMID- 15749266 TI - Early testicular effects in rats perinatally exposed to DEHP in combination with DEHA--apoptosis assessment and immunohistochemical studies. AB - This study aimed to characterize the effects of di(2-ethylhexyl) phthalate (DEHP) on the fetal rat testes and relate them to the effects seen in adults. Histopathological effects in fetal testes were examined with immunohistochemistry for anti-Mullerian hormone (AMH), 3beta-hydroxysteroid dehydrogenase, smooth muscle actin (SMA), proliferating cell nuclear antigen (PCNA), histone H3 and vimentin. Additionally, testicular apoptosis levels were assessed in fetal, prepubertal and adult rats. As the plasticizer di(2-ethylhexyl) adipate (DEHA) has similarities with DEHP in chemical structure and metabolism, we investigated if the testicular effects of DEHP were modulated by co-administration with DEHA. Wistar rats were gavaged during gestation and lactation with vehicle, DEHP (300 or 750 mg/kg/day), or DEHP (750 mg/kg/day) in combination with DEHA (400mg/kg/day), and male offspring were examined at gestation day (GD) 21, postnatal day (PND) 22, 26 and 190. In fetal testes, Leydig cells were found in large clusters containing AMH positive Sertoli cells. At GD 21, seminiferous chords appeared enlarged with an apparently increased number of gonocytes. However, proliferation of gonocytes did not appear increased. A few animals had a high number of TUNEL positive apoptotic cells in degenerating seminiferous tubules at PND 22 and 190, whereas most exposed animals had low levels of germ cell apoptosis at GD 21, PND 22 or PND 26, as evaluated by DNA laddering, TUNEL staining, Caspase-3 immunohistochemistry and Caspase-3 activity measurement. No differences between DEHP and DEHP+DEHA exposed groups were observed. PMID- 15749267 TI - Toxicological effects of acrylamide on rat testicular gene expression profile. AB - Toxicological effects of acrylamide on differential gene expression profile of rat testis were evaluated. Acrylamide induced morphological sperm defects, and decreased sperm concentration in cauda epididymis. Serum testosterone level and Leydig cell viability were also decreased dose-dependently, which resulted in decreased spermatogenesis. Acrylamide-induced histopathological lesions, such as formation of multinucleated giant cells and vacuolation, and numerous apoptotic cells were observed in seminiferous tubules. cDNA microarray analysis revealed that genes related to testicular-functions, apoptosis, cellular redox, cell growth, cell cycle, and nucleic acid-binding were up/down-regulated in testes isolated from acrylamide-treated group (60 mg/kg/day). Acrylamide toxicity appears to increase Leydig cell death and perturb gene expression levels, contributing to sperm defects and various abnormal histopathological lesions including apoptosis in rat testis. PMID- 15749268 TI - Multigenerational exposure to phytosterols in the mouse. AB - Due to beneficial health effects phytosterols (PS) are increasingly added to functional foods. The aim of the present study was to investigate the chronic effects of a dietary PS mixture (5mg/kg/day), containing mainly beta-sitosterol, on the reproduction of the mouse. General reproductive parameters, postnatal development, growth and survival of pups, weight of sex organs, the concentrations of plasma sex steroids and testicular testosterone were monitored across five generations (F(0)-F(4)). PS exposure increased the plasma levels of testosterone and decreased the relative uterine weights in the pups of F(2) and F(4) generations. Furthermore, PS exposure increased the concentrations of plasma estradiol in the female pups of F(3) generation. PS supplement also increased the testicular levels of testosterone in the male pups of F(2) generation. In spite of these transitory changes, PS exposure caused no permanent deleterious effects on the reproduction of the mouse. PMID- 15749269 TI - Reproductive and sexual behavior changes in male rats exposed perinatally to picrotoxin. AB - Previous studies in rats suggested that picrotoxin, a GABA(A) receptor antagonist, may cause long-term changes in male reproductive physiology and behavior in rats exposed during prenatal and postnatal periods. The present study has further examined this phenomenon. Wistar rat dams were dosed subcutaneously with 0.75 mg/kg picrotoxin in saline, or vehicle alone, during the perinatal period (day 19 of gestation, immediately after parturition, and once a day during the first 5 days of lactation). Birth weight and sexual maturation of pups were unchanged; however, plasma testosterone levels and sexual behavior was altered in male offspring. Although fertile, these males showed altered mating behavior in terms of a decrease in the mean number of mounts during a 30-min observation period with normal females. Some showed homosexual behavior when castrated and pretreated with exogenous estrogen. These findings suggest that perinatal exposure to picrotoxin alters sexual dimorphism in the developing rat brain, manifesting as altered reproductive performance and sexual behavior of males. PMID- 15749270 TI - Acute toxicity of textile dye wastewaters (untreated and treated) of Sanganer on male reproductive systems of albino rats and mice. AB - This study reports on the toxic effects of 15-days oral administration of untreated (Influent) and treated (Effluent) textile dye wastewaters on male reproductive systems of adult Swiss albino rats (age: 85-90 days) and mice (40-50 days). Textile dye wastewaters decreased body weight (7-25%) and reproductive organ weight (testis, epididymis, prostate gland and seminal vesicle, 2-48%). Similar trends were noted for total protein (14-70%), cholesterol (14-91%) and total lipid (10-30%) content of reproductive organs and spermatozoa, and for fructose levels in seminal vesicle (18-44%). Acid phosphatase activity in prostate however, was increased (11-44%) in the wastewater-exposed animals. Histopathological studies of treated animals revealed altered spermatogenesis, with higher sperm abnormalities, reduction in sperm counts (10-59%), and altered motility (14-56%). The magnitude of these abnormalities was stronger in rats versus mice, while among treatments, it was stronger in the Influent animals. Adverse effects improved when treated rats were allowed to recover 45 days in the control condition. Only recovered Effluent rats were capable of fertilizing normal females indicating that treated wastewater was less toxic; however, in comparison to control rats, litter size and body weight gains of F(1) and F(2) generations were adversely affected. Thus, the present study has established toxicity of both untreated and treated textile dye wastewater on reproductive biology of male Albino mice and rats. PMID- 15749271 TI - Pregnancy outcome after prenatal exposure to bleomycin, etoposide and cisplatin for malignant ovarian germ cell tumors: report of 2 cases. AB - Bleomycin plus etoposide and cisplatin seem to be a promising alternative for women with ovarian cancer. We are reporting two cases with favorable pregnancy outcome after exposure to these chemotherapeutic agents at the second and third trimesters of pregnancy. A pregnant woman with a stage Ic yolk-sac tumor underwent a right oophorectomy with omentectomy, and received five cycles of bleomycin, etoposide and cisplatin from the 22nd week of pregnancy until delivery. The second case was a pregnant woman with a stage Ia immature teratoma who underwent right salpingo-oophorectomy and received two cycles of bleomycin, etoposide and cisplatin starting at 30th week of pregnancy. The two patients did not have any evidence of recurrence of ovarian cancer for 6 and 2 years, respectively. Their babies did not have any evidence of minor or major malformations, and showed normal neurological development at 6 and 2 years of follow-up, respectively. PMID- 15749273 TI - Branching out into mix-and-match drug delivery. PMID- 15749274 TI - Surprise neuroprotective potential of beta-lactam antibiotics. PMID- 15749272 TI - A case of rosiglitazone exposure in the second trimester of pregnancy. AB - AIM: Data about rosiglitazone use in pregnancy is limited. We aimed to present a pregnant woman who exposed to rosiglitazone in the second trimester and the fetal outcome. SUBJECT: The case was a 42-year-old, multigravid Caucasian woman with a history of diabetes mellitus type II for 4 years prior to her current pregnancy. Her diabetes was managed by diet and exercise and she has not received any drug therapy until the 13th week of her sixth (present) and unplanned pregnancy. The case was exposed to rosiglitazone (4 mg/day) between 13th and 17th gestational weeks. After the diagnosis of pregnancy at the 17th week, rosiglitazone was stopped and insulin therapy was started. At the 37th week, she had a healthy male infant (4500 g, 50 cm). The baby was examined and no major or minor malformations were observed. CONCLUSION: This is the first case present in the literature exposed to rosiglitazone in the second trimester of her pregnancy. The data from the present case may contribute to the existing limited knowledge about rosiglitazone in pregnancy. PMID- 15749275 TI - Beyond PEGylation. PMID- 15749276 TI - Eric Kandel: a life in learning and memory. PMID- 15749277 TI - Obesity: the fat lady sings? PMID- 15749278 TI - Glycomics: coming of age across the globe. PMID- 15749279 TI - A soft approach to hard science? PMID- 15749280 TI - Mammalian carboxylesterases: from drug targets to protein therapeutics. AB - Our understanding of the detailed recognition and processing of clinically useful therapeutic agents has grown rapidly in recent years, and we are now able to begin to apply this knowledge to the rational treatment of disease. Mammalian carboxylesterases (CEs) are enzymes with broad substrate specificities that have key roles in the metabolism of a wide variety of clinical drugs, illicit narcotics and chemical nerve agents. Here, the functions, mechanism of action and structures of human CEs are reviewed, with the goal of understanding how these proteins are able to act in such a non-specific fashion, yet catalyze a remarkably specific chemical reaction. Current approaches to harness these enzymes as protein-based therapeutics for drug and chemical toxin clearance are described, as well as their uses for targeted chemotherapeutic prodrug activation. Also included is an outline of how selective CE inhibitors could be used as co-drugs to improve the efficacy of clinically approved agents. PMID- 15749281 TI - Discovery and development of biomarkers of neurological disease. AB - The identification of clinically relevant biomarkers for neurological diseases poses unique challenges. These include an historical lack of availability of relevant tissues from the site of pathology, relatively poorly matured techniques for disease diagnosis, the complexity and cellular heterogeneity of the brain, and a clear deficiency of models for functional validation of candidate biomarkers. Here, the unique challenges that neurological disorders introduce to biomarker discovery are described and how modern technological advances in genomics, proteomics and metabolomics are overcoming these obstacles and are driving the discovery of novel biomarkers to improve early diagnosis and therapeutic treatment is discussed. PMID- 15749282 TI - Cell culture-based models for intestinal permeability: a critique. AB - The model systems that are currently used to determine the intestinal permeability characteristics of discovery compounds often represent a combination of high-throughout, but less predictive, in silico and in vitro models and low throughput, but more predictive, in vivo models. Cell-based permeability models have been integrated into the discovery paradigm for some time and represent the "method of choice" across the industry. Here, in addition to an objective analysis of the utility of cell culture models for permeability screening, anticipated future trends in the field of cell culture models are discussed. PMID- 15749283 TI - From genome to antivirals: SARS as a test tube. AB - The severe acute respiratory syndrome (SARS) epidemic brought into the spotlight the need for rapid development of effective anti-viral drugs against newly emerging viruses. Researchers have leveraged the 20-year battle against AIDS into a variety of possible treatments for SARS. Most prominently, based solely on viral genome information, silencers of viral genes, viral-enzyme blockers and viral-entry inhibitors were suggested as potential therapeutic agents for SARS. In particular, inhibitors of viral entry, comprising therapeutic peptides, were based on the recently launched anti-HIV drug enfuvirtide. This could represent one of the most direct routes from genome sequencing to the discovery of antiviral drugs. PMID- 15749284 TI - Stable isotope methods for high-precision proteomics. AB - Stable isotope tagging methods provide a useful means of determining the relative expression level of individual proteins between samples in a mass spectrometer with high precision (coefficients of variation less than 10%). Because two or more samples tagged with different numbers of stable isotopes can be mixed before any processing steps, sample-to-sample recovery differences are eliminated. Mass spectrometry also allows post-translational modifications, splice variations and mutations (often unnoticed in immunoassays) to be detected and identified, increasing the clinical relevance of the assay and avoiding the issues of non specific binding and cross-reactivity observed in immunoassays. Several stable isotope tagging methods are available for use in proteomics research. We discuss the advantages and disadvantages of each technique with respect to biomarker discovery, target validation, efficacy and toxicology screening and clinical diagnostic applications. PMID- 15749285 TI - Global properties of biological networks. AB - This article discusses the most recent achievements in understanding the biological implications of the small-world and scale-free global topological properties of genetic, proteomic and metabolic networks. Most importantly, these networks are highly clustered and have small node-to-node distances. With their few very connected nodes, which are statistically unlikely to fail under random conditions, the proper functioning of these systems is maintained under external perturbations. PMID- 15749289 TI - Timeline of essential American College of Nurse-Midwives documents, clinical bulletins, and strategies for liability risk reduction. PMID- 15749290 TI - The profession of midwifery in the United States: Looking back and looking forward. PMID- 15749291 TI - Doubling back over roads once traveled: creating a national organization for nurse-midwifery. AB - The quest for a new national organization began in 1940 and concluded in November 1955 in Kansas City, Missouri, with the founding meeting of the American College of Nurse-Midwifery. This article looks at the conflicts with organized nursing about the place and role of nurse-midwives in the newly reorganized American Nurses Association and the National League for Nursing. Discussions and disagreements within nurse-midwifery over the need for a nonexclusive national organization that would set professional standards are examined. PMID- 15749292 TI - The emblem of the American College of Nurse-Midwives. PMID- 15749293 TI - Midwifery practice: Where have we been and where are we going? PMID- 15749294 TI - Presidential priorities: 50 years of wisdom as the basis of an action agenda for the next half-century. AB - Each of the living presidents of the American College of Nurse-Midwives (ACNM), past and current, was asked to select one or more issues that were of particular importance during her term(s) in office. Some of the issues identified by the presidents were amenable to review using existing quantitative data; this article presents those findings. The substantial increase in growth of ACNM membership in the 1980s and 1990s as well as the plateau that occurred at the beginning of the 21st century is documented. The relationship between ACNM and the professions of nursing, medicine, and other public health providers is illustrated. The increase in the number of Certified Nurse-Midwife/Certified Midwife (CNM/CM) education programs and the evolution and current profile of curriculum models are discussed. Finally, expansion of the scope of midwifery practice and the growing number of opportunities for entrepreneurial practice are presented in terms of current practice profiles, practice sites and settings, various employer-employee relationships, and the variety of financial reimbursement streams. This article is intended to complement the broader body of information about ACNM's history, and particularly, to supplement the richness of information derived from the qualitative and historical research inquiries conducted by others. PMID- 15749295 TI - The history and evolution of the Core Competencies for basic midwifery practice. AB - The Core Competencies in Nurse-Midwifery were first published in 1978 to provide a standard approach to nurse-midwifery education and now serve as the template for curriculum in nurse-midwifery/midwifery education. This article is an expansion of an earlier publication documenting the development of the Core Competencies document and describes its history and use in midwifery education and professional practice. The development and continued revision of this document are a significant part of ACNM and midwifery history in the United States. It defines the competencies essential to basic practice and can be used to define the scope of midwifery practice in both professional and policy arenas. PMID- 15749296 TI - Reflections on the past and future of midwifery research. PMID- 15749297 TI - Midwifery care of poor and vulnerable women, 1925-2003. AB - A systematic literature review of research on midwifery care of poor and vulnerable women from 1925 to 2003, which included topics studied, research methods used, and special issues and implications for future research, was performed; 44 studies published between 1955 and 2003 were identified. The majority were retrospective, descriptive studies. Outcomes examined included prenatal care visits, vaginal versus operative births, labor interventions, maternal and neonatal mortality and morbidity, birth weight, and cost effectiveness. Studies showed that midwives predominantly serve vulnerable women who are young, poor, immigrants, or members of racial and ethnic minorities. Preterm birth prevention is emerging as a midwifery research focus. Health system changes are making it more difficult to provide effective care and counseling to disadvantaged women, especially in managed care settings. Extensive evidence documents excellent outcomes of midwifery care for the poor in urban and rural settings over the past three quarters of a century. Future research should include more intervention studies and use both qualitative and quantitative methods to investigate midwifery processes of care and the process-outcome connection. The research focus should broaden beyond childbirth to include gynecology, family planning, and primary care issues. Health disparities, cultural studies, obstetric interventions, and poor women's experiences of childbirth and midwifery care are important topics for future research. PMID- 15749298 TI - Midwifery's research heritage: A Delphi survey of midwife scholars. AB - A Delphi survey was conducted with midwife scholars to determine the top 10 midwifery studies worthy of recognition in 2005, the 50th anniversary of the American College of Nurse-Midwives (ACNM). This survey was undertaken by students of Philadelphia University's Graduate Midwifery Program as a service-learning project on behalf of the ACNM Division of Research. Selected midwife scholars participated in 2 or 3 rounds of response and feedback to achieve consensus about research deemed historically or currently important to midwifery practice. The top 10 studies, as determined by the 19 participating midwife scholars, are presented here. These results are offered as reflection on research that has helped to shape and define the discipline of midwifery in the United States. PMID- 15749299 TI - The history of nurse-midwifery/midwifery education. PMID- 15749300 TI - The American College of Nurse-Midwives' dream becomes reality: The Division of Accreditation. AB - Recognized continuously by the US Department of Education since 1982 as a specialized accrediting agency, the American College of Nurse-Midwives' Division of Accreditation (DOA) accredits not only nurse-midwifery education programs at the postbaccalaureate or higher academic level as certificate and graduate programs for registered nurses (RNs), but also precertification programs for professional midwives from other countries who are licensed as RNs in the United States. The DOA also accredits midwifery education programs for non-nurses at the postbaccalaureate or higher academic level as certificate and graduate programs, and precertification programs for professional midwives from other countries. The accreditation process is a voluntary activity involving both nurse-midwifery and/or midwifery education programs and the DOA. Present plans include another expansion of recognition: to become an institutional accreditation agency for independent and proprietary schools and to continue as a programmatic accrediting agency. Since its inception, the accreditation process has been viewed as a positive development in nurse-midwifery education. PMID- 15749302 TI - The Journal of Midwifery & Women's Health 1955-2005: Its historic milestones and evolutionary changes. AB - From its very inception, the Journal of Midwifery & Women's Health (JMWH) played a critical role in fostering and documenting major shifts in the professional landscape of midwifery through its editorial content. This article traces the history and evolution of JMWH, commemorates its rich heritage and monumental growth over the last half century, highlights issues that have transcended its 50 year history, and acknowledges the contributions of its leadership ...past and present. PMID- 15749305 TI - Sebaceous carcinoma of the ocular region: a review. AB - Sebaceous carcinoma of the ocular region is a malignant neoplasm that is being recognized more frequently and managed by innovative techniques of local resection, cryotherapy, topical chemotherapy, and radiotherapy, resulting in improved visual and systemic prognosis. PMID- 15749306 TI - Amblyopia characterization, treatment, and prophylaxis. AB - Amblyopia has a 1.6-3.6% prevalence, higher in the medically underserved. It is more complex than simply visual acuity loss and the better eye has sub-clinical deficits. Functional limitations appear more extensive and loss of vision in the better eye of amblyopes more prevalent than previously thought. Amblyopia screening and treatment are efficacious, but cost-effectiveness concerns remain. Refractive correction alone may successfully treat anisometropic amblyopia and it, minimal occlusion, and/or catecholamine treatment can provide initial vision improvement that may improve compliance with subsequent long-duration treatment. Atropine penalization appears as effective as occlusion for moderate amblyopia, with limited-day penalization as effective as full-time. Cytidin-5' diphosphocholine may hold promise as a medical treatment. Interpretation of much of the amblyopia literature is made difficult by: inaccurate visual acuity measurement at initial visit, lack of adequate refractive correction prior to and during treatment, and lack of long-term follow-up results. Successful treatment can be achieved in at most 63-83% of patients. Treatment outcome is a function of initial visual acuity and type of amblyopia, and a reciprocal product of treatment efficacy, duration, and compliance. Age at treatment onset is not predictive of outcome in many studies but detection under versus over 2-3 years of age may be. Multiple screenings prior to that age, and prompt treatment, reduce prevalence. Would a single early cycloplegic photoscreening be as, or more, successful at detection or prediction than the multiple screenings, and more cost-effective? Penalization and occlusion have minimal incidence of reverse amblyopia and/or side-effects, no significant influence on emmetropization, and no consistent effect on sign or size of post-treatment changes in strabismic deviation. There may be a physiologic basis for better age-indifferent outcome than tapped by current treatment methodologies. Infant refractive correction substantially reduces accommodative esotropia and amblyopia incidence without interference with emmetropization. Compensatory prism, alone or post-operatively, and/or minus lens treatment, and/or wide-field fusional amplitude training, may reduce risk of early onset esotropia. Multivariate screening using continuous scale measurements may be more effective than traditional single-test dichotomous pass/fail measures. Pigmentation may be one parameter because Caucasians are at higher risk for esotropia than non-whites. PMID- 15749307 TI - Applications of liposomes in ophthalmology. AB - This review outlines the applications of liposomal formulations in ophthalmology. In ophthalmology, liposomes have been used to treat disorders of both the anterior and posterior segments. These include dry eyes, keratitis, corneal transplant rejection, uveitis, endophthalmitis, and proliferative vitreoretinopathy. Liposomes also have shown promise as vectors for genetic transfection and monoclonal antibody-directed vehicles. Furthermore, heat activated liposomes have spurred research in focal laser and heat-induced release of liposomal drugs and dyes for selective drug delivery. These techniques have been useful in selective tumor and neovascular vessel occlusion, angiography, and retinal and choroidal blood-flow studies. Although verteporfin is the only liposomal drug currently approved for use in the eye, the benefits of liposomes will likely be applied widely in all treatment, diagnostic, and research aspects of ophthalmology in the future. PMID- 15749309 TI - HMG CoA reductase inhibitors (statins): do they have a role in age-related macular degeneration? AB - Age-related macular degeneration is a progressive late onset disease affecting central vision. It is the leading cause of irreversible blindness in developed countries, and with the aging population the problem is increasing. Current treatment options are limited to the late stage of the disease when central vision is already under great threat, and even new treatments make little impact on the rate of blindness. Intervention earlier in the disease may prove more rewarding, but to date little progress has been made with this approach. Epidemiologic, genetic, and pathological evidence continues to accumulate, suggesting a possible link between risk factors for cardiovascular diseases and age-related macular degeneration. This article reviews the evidence and discusses the rationale behind the recent suggestions that cholesterol-lowering agents may be useful in the treatment of early age-related macular degeneration. The cholesterol-lowering family of drugs called statins are 3-hydroxy-3 methylglutaryl coenzyme A (HMG CoA) inhibitors with pleiotropic actions. Their therapeutic effects in cardiovascular disease and dyslipidaemia have been well proven. In this review we will outline the known actions of statins and discuss possible ways that they may impact on age-related macular degeneration. PMID- 15749308 TI - The macular xanthophylls. AB - The macular pigments are predominantly composed of three carotenoids: lutein, zeaxanthin, and meso-zeaxanthin. These carotenoids are concentrated and distributed in a selective manner. The properties of these pigments are further explored along with their methods of uptake, stabilization, and storage. The dual nature of these pigments as filters and antioxidants are elaborated upon in relation to their protective effects upon the macula, specifically in age-related macular degeneration. Evidence suggests that increased levels of macular pigment are correlated with a decreased risk of age-related macular degeneration. Many have sought to exploit this therapeutic relation. Studies reveal that oral supplementation with lutein and zeaxanthin can increase the levels of macular pigments in the retina and plasma. The effects of such supplementation on actual ocular function have yet to be fully addressed. New and standardized methods of assessing macular pigment density are discussed and future areas of research to further our understanding of macular xanthophylls as they pertain to age-related macular degeneration are highlighted. PMID- 15749310 TI - An eye for an eye; a tooth for a tooth. AB - A 24-year-old man experienced the sudden onset of a painless superior-temporal visual field defect of the left eye. Fundoscopy showed peripapillary pigmentary changes and a few nasal retinal white spots. Automated perimetry demonstrated an enlarged blind spot. The differential diagnosis of the various presumed inflammatory retinopathies and choroidopathies associated with an enlarged blind spot are reviewed and the classification of the white spot syndromes of the retina discussed. PMID- 15749311 TI - Ramon Castroviejo centenary: a life dedicated to corneal transplantation. AB - Ramon Castroviejo is widely recognized as one of the 10 most influential people in ophthalmology of the 20th century. Castroviejo was instrumental in the advancement of keratoplastic surgery, and his contributions as a researcher, surgeon, and teacher have benefited many. PMID- 15749312 TI - History of ophthalmology. Boerhaave's tale. AB - The great 18th century physician, Hermann Boerhaave, once lectured on a case that is most likely the second recorded description of transient blindness due to interference with the brain. PMID- 15749313 TI - Surveillance of hospital water and primary prevention of nosocomial legionellosis: what is the evidence? AB - Hospital-acquired Legionnaires' disease may be sporadic or may occur as part of an outbreak. As Legionella spp. are ubiquitous in many water systems, it is not surprising that hospital water may be colonized with Legionella pneumophila and other species. However, there is some controversy about the relationship between the presence of legionella in hospital water systems and nosocomial legionellosis. Primary prevention, i.e. measures to prevent legionella in a hospital or healthcare facility with no previous documented cases of nosocomial legionellosis, includes heightened awareness of hospital-acquired Legionnaires' disease with appropriate laboratory diagnostic facilities, and ensuring that the water system is well designed and maintained in accordance with national standards, e.g. the circulating hot water is maintained above 55 degrees C. Secondary prevention, i.e. preventing further cases occurring when a case has been confirmed, should include an investigation to exclude the hospital water system as a source. However, the necessity to sample hospital water routinely to detect legionella outside of outbreaks, i.e. as a component of primary prevention, is unclear. Some studies demonstrate a clear link but others do not. Differences between the patient populations studied, the methods of laboratory diagnosis of clinical cases, the analysis of hospital water and differences in the design of hospital water systems may partly explain this. Whilst further research, probably in the form of multi-centred prospective trials, is needed to confirm the relationship between environmental legionella and hospital-acquired legionellosis, including establishing the relative importance of L. pneumophila group 1 vs. non-group 1 and other Legionella spp., each hospital should consider the spectrum of patients at particular risk locally. Centres with transplant units or other patients with significant immunosuppression should, in the interim, consider routine sampling for legionella in hospital water in addition to other control measures. Therefore, infection control teams must work closely with hospital engineering and technical services departments and hospital management, as well as ensuring that physicians and others have a heightened awareness of hospital-acquired legionellosis. PMID- 15749314 TI - Comparison of low-temperature hydrogen peroxide gas plasma sterilization for endoscopes using various Sterrad models. AB - This study compared the effectiveness of sterilizing four types of endoscope using different models of the Sterrad system (Sterrad 50, 100, 100S and 200). Sterilization levels meeting international requirements were attained in all cases with carriers inoculated with Geobacillus stearothermophilus spores. The endoscopes were tested in half cycles ('overkill'). This is the first study to compare the Sterrad models marketed to date in terms of effective sterilization of endoscopes with narrow lumens. PMID- 15749315 TI - Intralaboratory reproducibility of the German test method of prEN ISO 15883-1 for determination of the cleaning efficacy of washer-disinfectors for flexible endoscopes. AB - In Europe, the evaluation of processing flexible endoscopes in washer disinfectors (WDs) is performed in accordance with prEN ISO 15883-1, which includes determination of cleaning efficacy. Recent data suggest that large differences are found between cleaning processes when the prEN ISO 15883-1 German test model is applied. Therefore, we analysed 12 separate cleaning processes, and each was assessed by determining the means of three experiments. Transparent test tubes used as test pieces (length 2m, lumen 2mm) were contaminated with a mixture of blood and Enterococcus faecium. The WD used was set to perform the cleaning process alone, which consists of three stages (pre-rinse, cleaning, intermediate rinse). Instead of a cleaning agent, demineralized water was used in all three stages of the cleaning process. The mean bacterial load was 10(11.3)cfu/mL (N=12) in the suspensions of test organism, 10(9.1)cfu/mL (N=12) in the test soils and 10(5.8)cfu/mL (N=12) in the controls (recovery). Mean log(10) reduction factors (RF) for each process, i.e. the difference in microbial loads between the control and the processed tubes, were calculated. The 12 cleaning processes with demineralized water led to a mean RF of 2.2 (range: 1.6-3.2) and excellent visible cleanliness of the test pieces. We found that the prEN ISO 15883-1 German test method leads to reproducible and valid results, and is suitable for determining the cleaning efficacy in WDs. PMID- 15749316 TI - Short antibiotic prophylaxis for bacterial infections in a neonatal intensive care unit: a randomized controlled trial. AB - We compared the effectiveness of a single dose and a three-day course of antibiotic prophylaxis in preventing bacterial infections in high-risk neonates. The study was a prospective, randomized controlled trial conducted in a 20-bed tertiary referral neonatal intensive care unit (NICU). A series of 130 neonates admitted consecutively to the NICU, fulfilling risk factors for infection, were assigned at random to receive intravenous antibiotic prophylaxis with ampicillin and netilmicin either in two daily doses for 72 h (three-day-administration group, 67 infants) or in a single bolus injection on admission (bolus group, 63 infants). Hospital-acquired infection, the main outcome measure, was defined as infection that developed at least 48 h after admission, and vertical infection (maternally transmitted) was considered to be present when clinical symptoms and abnormal laboratory findings became evident within 48 h of birth. Infections were considered as suspected when clinical and laboratory findings of infection were present, without positive cultures, and as confirmed when positive cultures were also present. No significant differences were found between the two groups of neonates studied in mean birth weight, gestational age or postnatal age on admission. The incidence of vertical infection was similar in the two groups (16/67, 23.9% vs. 14/63, 22.2%). Of the 130 newborns studied, 29 (22.3%) acquired at least one nosocomial infection during their NICU stay; total hospital-acquired infections, calculated as the incidence density of infection (the number of infective episodes divided by the number of days in the NICU), were less frequent among newborns who received the three-day course than the bolus (relative risk 0.69). This difference, although not statistically significant, depended on the different incidence density of confirmed nosocomial infections rather than on suspected infections (relative risk 0.59; 95% confidence interval 0.32-1.09; P=0.1). There were no significant differences between the two groups in overall mortality. A single bolus administration on admission is therefore likely to be as effective as a three-day course of antibiotic prophylaxis in preventing bacterial infection in high-risk infants admitted to an NICU. PMID- 15749317 TI - Systemic availability of prophylactic cefuroxime in patients submitted to coronary artery bypass grafting with cardiopulmonary bypass. AB - Cardiopulmonary bypass and hypothermia (HCPB) is a procedure commonly used during heart surgery, representing a risk factor for the patient by promoting extensive haemodilution and profound physiological changes. Cefuroxime is used for the prevention of infection following heart surgery, and several dose schemes have been suggested for prophylaxis with cefuroxime. The objective of the present study was to assess, in a comparative manner, the systemic availability of cefuroxime administered intravascularly as a bolus dose of 1.5 g to 17 patients having heart surgery with or without HCPB. Plasma cefuroxime concentrations were determined by high-pressure liquid chromatography-UV, and the following values, expressed as medians, were obtained for the study group compared with controls: 69.1 vs. 62.7 mg/L (1st h), 35.8 vs. 26.0mg/L (3rd h), 14.6 vs. 8.7 mg/L (6th h, P<0.05), 6.1 vs. 3.0mg/L (9th h, P<0.05) and 2.6 vs. 1.0mg/L (12th h, P<0.05). Despite the differences recorded during the study period as a consequence of HCPB, low antibiotic concentrations were found as early as 6h post dose for both groups investigated. Thus, the low systemic availability of cefuroxime after the administration of a 1.5-g dose may not protect against postoperative infections. The data obtained permit us to recommend a change in the dose scheme in order to maintain adequate plasma levels of cefuroxime. PMID- 15749318 TI - Surgical hand disinfection with a propanol-based hand rub: equivalence of shorter application times. AB - The aim of this study was to determine the efficacy of a propanol-based hand rub at application times shorter than 3 min. The bacterial pre-value was obtained from the finger tips (prEN 12791). Subjects treated their hands with the reference procedure (n-propanol, 60%) for 3 min or the product (crossover design). Sterillium was applied for 3, 2, 1.5 and 1 min. Four other preparations were tested for 1 min. Post-values (immediate effect) were taken from one hand, and the other hand was gloved for 3h. After the gloves were removed, the second post-value was taken (sustained effect). Sterillium was more effective than the reference procedure at 3, 2 and 1.5 min (immediate and sustained effect). The immediate effect after 1 min was significantly lower [mean log(10) reduction factor (RF): 1.91+/-0.90 vs. 2.52+/-0.95; P=0.001], whereas the sustained effect was not (mean RF: 1.81+/-1.06 vs. 2.05+/-1.14; P=0.204). All other preparations failed the efficacy requirement at 1 min for both the immediate and sustained effect. Using 2 x 3 mL Sterillium for a total of 1.5 min for surgical hand disinfection was at least as effective as the 3-min reference disinfection. PMID- 15749319 TI - Bacterial contamination of enteral nutrition in a paediatric hospital. AB - A prospective study was performed in a paediatric hospital to evaluate the incidence of bacterial contamination in enteral nutrition bags and to determine the critical points of process. During two separate one-month periods, all children receiving pump-assisted enteral nutrition were enrolled in the study. Samples for microbiological analysis were collected from enteral nutrition bags after administration in the first and second study period (sample T(2)). In the second study period, two additional samples were made at the end of the feed preparation process. One was refrigerated immediately (sample T(0)) and the other was sealed in a tube that followed the enteral nutrition solution until the end of its administration (sample T(1)). Bacterial contamination was detectable above 10(2)cfu/mL. Twenty-six out of 40 patients were included in the first study period and 14 out of 44 in the second study period. Contamination (>10(2)cfu/mL) occurred in nine of 26 samples (35%) and seven of 14 samples (50%) in the first and second study periods, respectively. Of these, five (20%) and three (21%) contained significant contamination (>/=10(4)cfu/mL). Bacteria of low pathogenicity were found in T(0) samples. Bacteria present in T(2) samples were pathogenic and multiple in 50% of cases. These results suggest that manipulation of the enteral nutrition bags at the bedside is critical for bacterial safety. PMID- 15749320 TI - Risk factors for ICU-acquired imipenem-resistant Gram-negative bacterial infections. AB - Intensive care units (ICUs) are high-risk areas for infections caused by antibiotic-resistant bacteria. This study investigated the risk factors for ICU acquired imipenem-resistant Gram-negative infections. It was conducted prospectively in three surgical ICUs and one medical ICU from April to December 2002. ICU-acquired Gram-negative infections were found in 128 patients. Of these, 42 had imipenem-resistant and 86 had imipenem-sensitive Gram-negative bacteria as the cause of infection. According to the univariate analysis results, hospital stay before ICU admission, hospitalization period before ICU admission, length of ICU stay, surgical ICU stay, surgical operation and previous antibiotic use were significant risk factors for the acquisition of imipenem-resistant infections. In the multivariate analysis, length of ICU stay, surgical operation and previous carbapenem use were independently associated with imipenem resistance. PMID- 15749321 TI - Relationship between nosocomial infection and mortality in a neurology intensive care unit in Turkey. AB - Few previous studies have evaluated the relationship between nosocomial infection and mortality in a neurology intensive care unit (ICU). In this study, patients treated for more than 24h in the neurology ICU of the Ankara Training and Research Hospital, Turkey were followed until death or two days after discharge by prospective daily surveillance. The study period was 14 months. One hundred and sixty-nine ICU-acquired infections occurred in 74 (38.9%) of 190 patients during 2006 patient-days. The overall rate of ICU-acquired nosocomial infection was 88.9/100 patients and 84.2/1000 patient-days. While the overall mortality rate was 60%, mortality in patients with nosocomial infections was 69%. In univariate analysis, infection (nosocomial and community-acquired) (P=0.002), nosocomial infection (P<0.05), mechanical ventilation (P<0.0001), presence of two or more underlying diseases (P=0.01), parenteral nutrition (P<0.0001), steroid treatment (P=0.003) and a low Glasgow Coma Scale (GCS) score (P=0.0001) were identified as risk factors for mortality. Stepwise logistic regression analysis showed nosocomial infection (P<0.05), mechanical ventilation (P=0.009), the presence of two or more underlying diseases (P<0.05) and a low GCS score (P=0.0001) to be risk factors for ICU mortality. It was concluded that nosocomial infection increases the risk of mortality by a factor of 1.69. The impact of nosocomial infection on mortality in our ICU was higher in patients with high GCS scores and patients aged between 66 and 75 years. In particular, nosocomial infection increased mortality among patients with less severe illnesses. PMID- 15749322 TI - Increased risk of bloodstream and urinary infections in intensive care unit (ICU) patients compared with patients fitting ICU admission criteria treated in regular wards. AB - Critically ill patients, eligible for admission into intensive care units (ICUs), are often hospitalized in other wards due to a lack of ICU beds. Differences in morbidity between patients managed in ICUs and elsewhere are unknown, specifically the morbidity related to hospital-acquired infection. Patients fitting ICU admission criteria were identified by screening five entire hospitals on four separate days. Hospital infections within a 30-day follow-up period were compared in ICU patients and in patients on other wards using Kaplan-Meier curves. Residual differences in the patients' case mix between ICUs and other wards were adjusted for utilizing multivariate Cox models. Of 13415 patients screened, 668 were critically ill. The overall infection rates (per 100 patient days) were 1.2 for bloodstream infection (BSI) and 1.9 for urinary tract infection (UTI). The adjusted hazard ratios in ICU patients compared with patients on regular wards were 3.1 (P<0.001) for BSI and 2.5 (P<0.001) for UTI. This increased risk persisted even after adjusting for the disparity in the number of cultures sent from ICUs compared with ordinary wards. No interdepartmental differences were found in the rates of pneumonia, surgical wound infections and other infections. Minimizing the differences between characteristics of patients hospitalized in ICUs and in other wards, and controlling for the higher frequency of cultures sent from ICUs did not eliminate the increased risk of BSI and UTI associated with admission into ICUs. PMID- 15749323 TI - Prevalence of hospital-acquired infection in a Tunisian hospital. AB - In order to estimate the prevalence of hospital-acquired infection (HAI) and research factors associated with its occurrence, a one-day prevalence survey was conducted at the Habib Bourguiba University Hospital, Tunisia. We studied 280 patients who had been present in the same ward for at least 48 h, and who had occupied a hospital bed between 17 April 2002 (midnight) and 18 April 2002 (midnight). The overall prevalence of HAI was 17.9%. The most frequently infected sites were the lungs (32%), surgical wounds (28%) and the urinary tract (20%). Microbiological documentation was available in only 28% of HAIs, and the most frequently isolated organisms were Gram-negative rods (80.8%). Results of multiple logistic regression analysis indicated that HAI is linked to the medical category, the use of intravascular devices and antibiotic prophylaxis. This survey provided information on the prevalence of HAI in Tunisian hospitals, the breakdown of infections, and HAI predisposing factors. PMID- 15749324 TI - Bacterial contamination of dental chair units in a modern dental hospital caused by leakage from suction system hoses containing extensive biofilm. AB - Within six months of opening of the new Dublin Dental Hospital in September 1998, areas of corrosion were observed on many of the baseplates of the hospital's 103 dental chair units (DCUs) at the site of attachment of the suction hoses. The corroded areas were heavily contaminated with Pseudomonas spp. and related genera posing a risk of cross-infection, particularly for immunocompromised patients. These species were used as marker organisms to investigate the source of the contamination. P. aeruginosa was the predominant species recovered from 41 selected DCU baseplates (61% prevalence), whereas P. putida (46% prevalence) and P. aeruginosa (43% prevalence) were predominant at the attachment ends of 37 selected high-volume suction hoses. Forty-one selected isolates of P. aeruginosa from 13 DCU baseplates, 16 high-volume suction hoses and 12 coarse filter housings (another suction system site) from 19 separate DCUs were serotyped to determine the similarity of isolates at each site. The majority of isolates (68.3%) belonged to serotype O:10, while the remainder belonged to serotypes O:6 (7.3%), O:11 (7.3%), O:14 (9.8%) and O:5/O:16 (7.3%). Of the isolates from DCU baseplates, additional isolates with the same serotype were recovered from other suction system sites in 10/13 (77%) cases. Isolates of only one serotype were recovered from each of the 19 DCUs investigated. Forty-one serotyped isolates were also subject to computer-assisted analysis of SpeI-generated DNA fingerprint profiles, and similarity coefficient (S(AB)s) values were calculated for each pairwise combination of isolate profiles. The data obtained showed that the isolates consisted of two distinct main populations, each containing separate clades corresponding to specific serotypes. Serotype O:6 (three isolates), O:11 (three isolates) and O:5/O:16 (three isolates) belonged to a single strain in each case. Serotypes O:14 (four isolates) and O:10 (28 isolates) belonged to two strains in each case. The two serotype O:10 strains, termed fingerprint groups I (four isolates from three DCUs) and II (24 isolates from 10 DCUs), were the most distantly related of all the strains identified. These findings demonstrated that the hospital DCUs had become colonized with a small number of P. aeruginosa strains, one of which (serotype O:10, fingerprint group II) predominated. These results also confirmed that DCU baseplate contamination was most likely to be due to leakage from suction system hoses at the baseplate attachment sites, probably due to loosening during use. Replacement hose connectors that firmly retained the suction hoses in the attachment sites so that they could not be loosened by movement of the suction hoses solved this problem, and eliminated further contamination of the DCU baseplates. PMID- 15749325 TI - Residual formaldehyde on plastic materials and medical equipment following low temperature steam and formaldehyde sterilization. AB - We measured the amount of residual formaldehyde on 16 plastic materials and five medical devices following low-temperature steam and formaldehyde (LTSF) sterilization, based on the European Standard EN14180. The amounts of formaldehyde residue on the plastic materials were compared with that on a filter paper of similar dimensions. The amount of residual formaldehyde on polyamide 6, polyurethane, natural rubber and polyacetal was higher (21.9, 15.2, 3.0 and 2.1 times, respectively) than that on the filter paper. The amount of formaldehyde recovered from a breathing circuit, anaesthesia circuit, oxygen tubing, airway tube and tweezers was 260, 240, 594, 56 and 0 microg, respectively, following LTSF sterilization. Our results emphasize the need to verify the main material composing the medical equipment before LTSF sterilization, as the amount of formaldehyde retrieved following sterilization varies according to the material used for construction. PMID- 15749326 TI - Protecting healthcare staff from severe acute respiratory syndrome: filtration capacity of multiple surgical masks. AB - Guidelines issued by the Centers for Disease Control and Prevention and the World Health Organisation state that healthcare workers should wear N95 masks or higher level protection during all contact with suspected severe acute respiratory syndrome (SARS). In areas where N95 masks are not available, multiple layers of surgical masks have been tried to prevent transmission of SARS. The in vivo filtration capacity of a single surgical mask is known to be poor. However, the filtration capacity of a combination of masks is unknown. This was a crossover trial of one, two, three and five surgical masks in six volunteers to determine the in vivo filtration efficiency of wearing more than one surgical mask. We used a Portacount to measure the difference in ambient particle counts inside and outside the masks. The best combination of five surgical masks scored a fit factor of 13.7, which is well below the minimum level of 100 required for a half face respirator. Multiple surgical masks filter ambient particles poorly. They should not be used as a substitute for N95 masks unless there is no alternative. PMID- 15749327 TI - Hand decontamination by medical staff in general medical wards. PMID- 15749328 TI - Hand-hygiene behaviour, attitudes and beliefs in first year clinical medical students. PMID- 15749329 TI - The epic project: updating the evidence base for national evidence-based guidelines for preventing healthcare-associated infections in NHS hospitals in England. A report with recommendations. PMID- 15749331 TI - Frequency of replacing peripheral venous catheters. PMID- 15749330 TI - Microbial mutation as a source of polyclonality in the gut of the critically ill. PMID- 15749332 TI - Photoperiod alters hypothalamic cytokine gene expression and sickness responses following immune challenge in female Siberian hamsters (Phodopus sungorus). AB - Rodents that live in changing environments display different immune responses mediated in part by photoperiod (day length) cues. Siberian hamsters maintained in winter-like (short) photoperiods display smaller physiological and behavioral responses to immune challenges as compared with hamsters housed in summer-like (long) photoperiods. We hypothesized that these different response patterns are attributable to altered cytokine production in the hypothalamus in response to photoperiod changes. Female hamsters were housed in long or short days for 10 weeks to induce photoperiodic alterations, then injected with either LPS (a bacterial endotoxin) or saline. Fever and food intake were assessed 3 h post injection; hypothalami and blood were collected 3, 6, and 12 h post-injection. LPS induced lower fever and reduction in food intake responses in short-day hamsters as compared with long-day hamsters. Additionally, short-day hamsters reduced IL-1beta and Tnfalpha expression in the hypothalamus 6 h after LPS injection, as measured by quantitative RT-PCR. Plasma estradiol concentrations did not differ between long- and short-day hamsters. These data suggest that differences in cytokine production in the hypothalamus may underlie the photoperiod-induced differences in sickness responses, and that these changes are not mediated by estradiol. PMID- 15749334 TI - Induction of long-term potentiation leads to increased reliability of evoked neocortical spindles in vivo. AB - Large amplitude electroencephalographic spindle waves (7-14 Hz) occur spontaneously in the neocortex during both sleep and awake immobility, and it has been proposed that synchronous neuronal activation during spindles may contribute to learning-related synaptic plasticity. Spindles can also be evoked in the sensorimotor cortex by electrical stimulation of cortical or thalamic inputs in the rat. To determine if strengthening cortical synapses can affect the initiation and maintenance of electrically evoked spindles, stimulation pulses were delivered at a range of intensities to the corpus callosum or ventrolateral thalamus in the awake rat before and after the induction of long-term potentiation (LTP) by tetanization of the corpus callosum. The morphology of evoked spindles was similar to that of naturally occurring spindles. Spindles were evoked less reliably during slow-wave sleep than during waking, and this was correlated with smaller synaptic responses during slow-wave sleep. Similar to previous findings, daily tetanization of the corpus callosum for 15 days decreased the early component and increased the late component of synaptic responses evoked by corpus callosum stimulation, but did not significantly affect synaptic responses evoked by thalamic stimulation. Similarly, LTP induction increased the reliability with which low-intensity corpus callosum stimulation evoked spindles, but increases in spindles evoked by thalamic stimulation were not significant. Synaptic potentiation and the increased reliability of spindles developed with a similar time-course over the 15-day LTP induction period. These results reflect strong correlations between the strength of cortical layer V activation and the initiation of spindles in the sensorimotor cortex, and support the idea that monosynaptic and polysynaptic horizontal collaterals of layer V neurons can play a significant role in the initiation of spindles. PMID- 15749333 TI - Neuro-overprotection? A functional evaluation of clomethiazole-induced neuroprotection following hypoxic-ischemic injury. AB - Hypoxic-ischemic (H-I) injury produces extensive damage to the hippocampus of young rats. We have recently shown that administration of 125 mg kg-1 clomethiazole (CMZ), a GABA(A)-agonist, provides complete histological protection against H-I injury if administered 3 h post-H-I (Brain Res 1035 (2005) 194). However, whether that histological protection translates into lasting functional preservation is unclear. To determine whether hippocampal-based circuits remain functionally intact in CMZ-protected H-I rats, we administered 125 mg kg-1 (high dose [CMZ-HD]) or 65 mg kg-1 (low dose [CMZ-LD]) CMZ, 3 h post-H-I, and examined numerous kindling parameters in the dorsal hippocampus 60 days following H-I. Kindling parameters included afterdischarge (AD) thresholds (ADTs), AD durations and kindling rates. Additional groups assessed included vehicle-injected H-I (VIH), hypoxic, ligated and naive rats. VIH, CMZ-HD, CMZ-LD and hypoxic rats all exhibited significantly faster kindling rates than naive rats. Thus, a previous traumatic event, even hypoxia alone, facilitated subsequent seizure propagation. Still, a significantly slower kindling rate was evident in CMZ-HD rats than in hypoxic, VIH or CMZ-LD rats. Moreover, while longer pre-kindling AD durations were observed in the damaged hippocampus of VIH compared with naive rats, this was not true for either CMZ-treated groups, hypoxic or ligated rats. Collectively, these findings suggest CMZ can suppress the epileptogenic effects of H-I. Surprisingly, however, both groups of CMZ-treated rats exhibited a four to nine times greater ADT than any other group and this effect was most profound in the CMZ-protected hippocampus. Thus, CMZ administration protected local neurons against terminal insult and left network excitability relatively normal with respect to seizure offset mechanisms but also caused profound elevation of local ADTs, which suggests a local hypoexcitability/increased inhibition. Finally, this study demonstrates, for the first time, that the kindling model can serve as a sensitive measure of function-related neuroprotective efficacy in animal models of ischemia. PMID- 15749335 TI - N-methyl-D-aspartic acid-induced and Ca-dependent neuronal swelling and its retardation by brain-derived neurotrophic factor in the epileptic hippocampus. AB - Dentate granule cell (DGC) swelling was studied by imaging changes in light transmittance from hippocampal slices in the rat pilocarpine model of epilepsy and human epileptic specimens. Brief bath-application of N-methyl-D-aspartic acid (NMDA) induced swelling in the control rat DGC (physiological swelling). Physiological swelling was short-lasting, and rapidly recovered upon removal of NMDA. In contrast, the swelling induced in the pilocarpine-treated rat hippocampus and human epileptic hippocampus (epileptic swelling) was long lasting, and often recovered slowly over an hour. Both types of swelling were blocked by the NMDA receptor (NMDAR) antagonist, D-APV, suggesting that they shared the same induction mechanism. However, the swellings differed in their sensitivity to a calcium chelator, 1.2-bis(2-aminophenoxy)ethane-N,N,N,N-tetra acetate (BAPTA), and an endoplasmic reticulum (ER) Ca2+-ATPase inhibitor, thapsigargin (TG). BAPTA and TG affected only epileptic swelling, and physiological swelling was spared. This suggested that the NMDAR-induced epileptic swelling might involve an additional mechanism for its maintenance, likely recruiting ER Ca2+ stores. Brain-derived neurotrophic factor (BDNF) slightly attenuated physiological swelling, and blocked epileptic swelling. The present study suggests a functional link between the activation of NMDAR and a release of Ca2+ from internal stores during the induction of epileptic swelling, and a neuroprotective role of BDNF on the NMDAR-induced swelling in the epileptic hippocampus. PMID- 15749336 TI - Analysis of SNAP-25 immunoreactivity in hippocampal inhibitory neurons during development in culture and in situ. AB - Synaptosomal associated protein of 25 kDa (SNAP-25) is a component of the soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor (SNARE) complex which plays a central role in synaptic vesicle exocytosis. We have previously demonstrated that adult rat hippocampal GABAergic synapses, both in culture and in brain, are virtually devoid of SNAP-25 immunoreactivity and are less sensitive to the action of botulinum toxin type A, which cleaves this SNARE protein [Neuron 41 (2004) 599]. In the present study, we extend our findings to the adult mouse hippocampus and we also provide demonstration that hippocampal inhibitory synapses lacking SNAP-25 labeling belong to parvalbumin-, calretinin- and cholecystokinin-positive interneurons. A partial colocalization between SNAP 25 and glutamic acid decarboxylase is instead detectable in developing mouse hippocampus at P0 and, at a lesser extent, at P5. In rat embryonic hippocampal cultures at early developmental stages, SNAP-25 immunoreactivity is detectable in a percentage of GABAergic neurons, which progressively reduces with time in culture. Consistent with the presence of the substrate, botulinum toxin type A is partially effective in inhibiting synaptic vesicle recycling in immature GABAergic neurons. Since SNAP-25, beside its role as a SNARE protein, is involved in additional processes, such as neurite outgrowth and regulation of calcium dynamics, the presence of higher levels of the protein at specific stages of neuronal differentiation may have implications for the construction and for the functional properties of brain circuits. PMID- 15749337 TI - Metabotropic glutamate receptor/phospholipase C pathway: a vulnerable target to Creutzfeldt-Jakob disease in the cerebral cortex. AB - Glutamate is the main excitatory neurotransmitter in the cerebral cortex. Altered glutamatergic transmission has been suggested as having a central role in many neurodegenerative diseases. Metabotropic glutamate receptors (mGluRs) are coupled to intracellular signal transduction via G proteins, and they mediate slower responses than ionotropic glutamate receptors. Group I mGluRs are positively coupled to phospholipase C beta1 (PLCbeta1). Creutzfeldt-Jakob disease (CJD) is a human transmissible spongiform encephalopathy associated with a dysfunction in the membrane glycoprotein PrP which is converted into an abnormal isoform, with a predominant beta-sheet structure, that is pathogenic and partially resistant to protease digestion. Proteins associated with the signal transduction of group I mGluRs were examined in the frontal cortex (area 8) of 12 cases with sCJD and four age-matched controls, by means of gel electrophoresis and Western blotting of total homogenates. Densitometric analysis of the bands demonstrated decreased expression levels of PLCbeta1 and PLCgamma, a non-related phospholipase which is a substrate of tyrosine kinase, in CJD cases when compared with controls. Novel protein kinase C delta (nPKCdelta) has also been found to be significantly decreased in CJD cases. However, no modifications in mGluR1 cPKCalpha expression levels are found in CJD when compared with controls. No modifications in PLCbeta1 solubility in PBS-, deoxycholate- and sodium dodecylsulphate-soluble fractions have been observed in CJD when compared with controls. Finally, no interactions between PLCbeta1 and PrP, as revealed by immunoprecipitation assays, have been found in CJD and controls. The present results show, for the first time, reduced expression levels of phospholipases, particularly PLCbeta1, which may interfere with group I mGluR signaling in the cerebral cortex in CJD. These abnormalities are not the result of abnormal PLC solubility or interactions with PrP. Selective involvement of group I mGluRs may have functional effects on glutamatergic transmission modulation and processing in CJD. PMID- 15749338 TI - Ectopic action potential generation in cortical interneurons during synchronized GABA responses. AB - In the presence of 4-aminopyridine and excitatory amino acid receptor antagonists, individual neurons in brain slice preparations exhibit large gamma aminobutyric acid (GABA)-mediated responses as a consequence of synchronous GABA release from a network of interneurons. These synchronized GABA responses are frequently associated with ectopic action potentials (EAPs), which are thought to be action potentials initiated in distal axon terminals which subsequently travel antidromically toward the soma. Ectopic action potentials feature prominently in some models of epilepsy. Neocortical synchronized GABA responses propagate across the cortex, predominantly in superficial layers. The role that EAPs may play in contributing to laminar differences in the synchronized GABA response has not been addressed. Here we examined the occurrence of EAPs during synchronized GABA responses in neurons within layers I and II/III. EAPs occurred in 78% of layer I interneurons and in 25% of layer II/III interneurons (including chandelier cells). EAPs were not observed in layer II/III pyramidal neurons. The prevalence of EAPs in layer I interneurons provides a mechanism by which layer I can support both the initiation and propagation of synchronized GABA responses. Thus, layer I interneurons are a critical component of a network capable of synchronizing a propagating wave of GABA release across the neocortex. PMID- 15749339 TI - Neuronal intranuclear inclusions and neuropil aggregates in HdhCAG(150) knockin mice. AB - We studied the development of neuronal intranuclear inclusions (NIIs), neuropil aggregates (NAs), and expression of expanded repeat polyglutamine protein in the HdhCAG(150) knockin mouse model of Huntington's disease (HD). Diffuse nuclear localization of huntingtin protein (htt) was noted initially within striatal neurons at approximately 28 weeks, followed by the development of striatal htt immunoreactive NIIs by approximately 40 weeks. Striatal NIIs were observed initially in clusters within the matrix compartment but subsequently became diffusely distributed throughout the striatum. In the oldest animals (107 weeks), NIIs were enlarged and diffuse nuclear htt immunoreactivity reduced. Expression of ubiquitin immunoreactive NIIs paralleled but lagged behind the expression of htt immunoreactive NIIs. Abundant NIIs were found by approximately 75 weeks in layers 3 and 4 of somatosensory cortex and in layer 2 of piriform cortex. In the oldest animals, greater than 100 weeks, some NIIs were found in many brain regions. NAs were found mainly within the globus pallidus and substantia nigra, perhaps reflecting expression in striatal terminals. Cyclic AMP response element binding protein (CBP) was not localized to NIIs, arguing against gross sequestration of this transcriptionally active protein. Comparison of the relative levels of a common polyglutamine epitope in HdhCAG(150) knockin and hprtCAG(146) knockin mice shows greater expression of the polyglutamine epitope in the phenotypically less aggressive HdhCAG(150) knockin line. HdhCAG(150) knockin mice may be a model of early pathologic changes in HD. PMID- 15749340 TI - Genetic background regulates semaphorin gene expression and epileptogenesis in mouse brain after kainic acid status epilepticus. AB - The host response to neural injury, which can include axonal sprouting and synaptic reorganization is likely to be under tight genetic regulatory control at the level of the genome and may be implicated in epileptogenesis. Despite its importance, however, the molecular basis of synaptic reorganization is unclear. We have studied the development of synaptic reorganization, semaphorin gene expression, and epileptogenesis in hippocampus of epileptogenic sensitive (FVB/NJ) and epileptogenic resistant (C57BL/6J) mice (i.e. distinct genetic backgrounds) after kainic acid-induced status epilepticus. Our results support the hypothesis that disruption of transcriptional regulation of axon guidance genes leads to a differential loss of tonic neuropilin-2 dependent activation of semaphorin 3F receptors on hippocampal neurons on distinct genetic backgrounds. This results in rearranged synaptic circuitry and thus promotes epileptogenesis. These findings may define biologic principles underlying the role of semaphorin signaling which may broadly apply to other systems undergoing neural regeneration. PMID- 15749341 TI - Effect of polymorphism on expression of the neuropeptide Y gene in inbred alcohol preferring and -nonpreferring rats. AB - Using animal models of alcoholism, previous studies suggest that neuropeptide Y (NPY) may be implicated in alcohol preference and consumption due to its role in the modulation of feeding and anxiety. Quantitative trait loci (QTL) analysis previously identified an interval on rat chromosome 4 that is highly associated with alcohol preference and consumption using an F2 population derived from inbred alcohol-preferring (iP) and -nonpreferring (iNP) rats. NPY mapped to the peak of this QTL region and was prioritized as a candidate gene for alcohol seeking behavior in the iP and iNP rats. In order to identify a potential mechanism for reduced NPY protein levels documented in the iP rat, genetic and molecular components that influence NPY expression were analyzed between iP and iNP rats. Comparing the iP rat to the iNP rat, quantitative real-time polymerase chain reaction detected significantly decreased levels of NPY mRNA expression in the iP rat in the six brain regions tested: nucleus accumbens, frontal cortex, amygdala, hippocampus, caudate-putamen, and hypothalamus. In addition, the functional significance of three previously identified polymorphisms was assessed using in vitro expression analysis. The polymorphism defined by microsatellite marker D4Mit7 in iP rats reduced luciferase reporter gene expression in SK-N-SH neuroblastoma cells. These results suggest that differential expression of the NPY gene resulting from the D4mit7 marker polymorphism may contribute to reduced levels of NPY in discrete brain regions in the iP rats. PMID- 15749342 TI - Amyloid-beta-stimulated plasminogen activation by tissue-type plasminogen activator results in processing of neuroendocrine factors. AB - Alzheimer's disease brain is characterized by the abundant presence of amyloid deposits. Accumulation of the major constituent of these deposits, amyloid-beta (Abeta), has been associated with decreased neurotransmission, increased neuronal cell death, and with cognitive decline. The mechanisms underlying these phenomena have not yet been fully elucidated. We have previously shown that amyloid peptides like Abeta bind tissue-type plasminogen activator (tPA) and cause enhanced plasmin production. Here we describe the identification of five major neuronal cell-produced Abeta-associated proteins and how Abeta-stimulated plasmin formation affects their processing. These five proteins are all neuroendocrine factors (NEFs): chromogranins A, B and C; truncated chromogranin B; and VGF. Plasminogen caused processing of Abeta-bound (but not soluble) tPA, chromogranin B and VGF and the degradation products were released from Abeta. Processing of the neuroendocrine factors was dependent on tPA as it was largely abrogated in tPA-/- cells or in the presence of a specific tPA-inhibitor. If plasmin indeed produces NEF-derived peptides in vivo, some of these peptides may have biological activity, for instance in regulating neurotransmitter release that may affect the pathology of Alzheimer's disease. PMID- 15749343 TI - Caspase 6 expression in the rat hippocampus during epileptogenesis and epilepsy. AB - The molecular basis of neuronal circuit reorganization during epileptogenesis is poorly understood. Such data are, however, critical for the search of new targets for the prevention of epileptogenesis. Here, we extended our previous studies on caspases in epileptogenesis by investigating the expression and activity of caspase 6 at different phases of the epileptic process in rats. Epileptogenesis was triggered by kainate-induced status epilepticus (SE) under video electroencephalography-monitoring. Caspase 6 activity was measured fluorometrically in the hippocampus 8 h, 24 h, 48 h, 1 week, and 4 weeks after SE. Caspase 6 expression was examined using Western blot and immunohistochemistry. Our data demonstrated that the SE-induced increase in the expression of cleaved caspase 6 and its intraneuronal localization were dependent on the time delay from SE induction. Double-labeling with a neuronal marker, NeuN, indicated that within the first 48 h, caspase 6 immunoreactivity was present both in the hippocampal pyramidal cells and hilar neurons, some of which were also terminal transferase dUTP-end labeling-positive. This was coincident with a transient 18-fold increase in caspase 6 enzymatic activity. At the 1-week and 4-week time points, elevated caspase 6 immunoreactivity was detected in the dendritic processes and neuropil. These findings indicate that caspase 6 expression remains elevated long after the occurrence of acute cell death during epileptogenesis and epilepsy. Further, caspase 6 protein is not exclusively located in the somata of neurons, but also in dendrites. These data suggest that caspase 6 has functions other than execution of programmed cell death in epileptogenic hippocampus. PMID- 15749344 TI - Neuroprotective properties of cultured neural progenitor cells are associated with the production of sonic hedgehog. AB - Numerous studies have shown that abnormal motor behavior improves when neural progenitor cells (NPCs) are transplanted into animal models of neurodegeneration. The mechanisms responsible for this improvement are not fully understood. Indirect anatomical evidence suggests that attention of abnormal motor behavior is attributed, at least in part, to the secretion of trophic factors from the transplanted NPCs. However, there is little direct evidence supporting this hypothesis. Here we show that NPCs isolated from the subventricular zone (SVZ) of neonatal mice are highly teratogenic when transplanted into the neural tube of developing chick embryos and are neuroprotective for fetal dopaminergic neurons in culture because they release sonic hedgehog (Shh). In addition, the neuroprotective properties of NPCs can be exploited to promote better long-term survival of transplanted fetal neurons in an animal model of Parkinson's disease. Thus, cultured NPCs isolated from the SVZ can secrete at least one potent mitogen (Shh) that dramatically affects the fate of neighboring cells. This trait may account for some of the improvement in motor behavior often reported in animal models of neurodegeneration after transplantation of cultured NPCs that were isolated from the SVZ. PMID- 15749345 TI - Synaptic and extrasynaptic GABA-A and GABA-B receptors in the globus pallidus: an electron microscopic immunogold analysis in monkeys. AB - GABA-A and GABA-B receptors mediate differential effects in the CNS. To better understand the role of these receptors in regulating pallidal functions, we compared their subcellular and subsynaptic localization in the external and internal segments of the globus pallidus (GPe and GPi) in monkeys, using pre- and post-embedding immunocytochemistry with antibodies against GABA-A (alpha1, beta2/3 subunits) and GABA-BR1 receptor subtype. Our results demonstrate that GABA-A and GABA-B receptors display a differential pattern of subcellular and subsynaptic localization in both segments of the globus pallidus. The majority of GABA-BR1 immunolabeling is intracellular, whereas immunoreactivity for GABA-A receptor subunits is mostly bound to the plasma membrane. A significant proportion of both GABA-BR1 and GABA-A receptor immunolabeling is extrasynaptic, but GABA-A receptor subunits also aggregate in the main body of putative GABAergic symmetric synapses established by striatal- and pallidal-like terminals. GABA-BR1 immunoreactivity is expressed presynaptically in putative glutamatergic terminals, while GABA-A alpha1 and beta2/3 receptor subunits are exclusively post-synaptic and often coexist at individual symmetric synapses in both GPe and GPi. In conclusion, our findings corroborate the concept that ionotropic and metabotropic GABA receptors are located to subserve different effects in pallidal neurons. Although the aggregation of GABA-A receptors at symmetric synapses is consistent with their role in fast inhibitory synaptic transmission, the extrasynaptic distribution of both GABA-A and GABA-B receptors provides a substrate for complex modulatory functions that rely predominantly on the spillover of GABA. PMID- 15749346 TI - Decreased glutathione transferase levels in rd1/rd1 mouse retina: replenishment protects photoreceptors in retinal explants. AB - Currently much attention is focused on glutathione S transferase (GST)-induced suppression of apoptosis. The objective of our studies was therefore to see if GST isoenzymes rescue photoreceptors in retinal explants from rd1/rd1 mice, in which photoreceptors degenerate rapidly. Eyes from C3H rd1/rd1 and +/+ mice were collected at various time points between postnatal day (PN) 2 and PN28. Localization and content of alpha-GST and mu-GST was investigated by immunofluorescence and semi-quantitative Western blot analysis, respectively. In addition, PN2 and PN7 retinal explants were cultured till PN28, during which they were treated with 10 ng/ml alpha-GST or mu-GST. The spatiotemporal expression of both GST isoforms was closely similar: early presence in ganglion cell layer after which staining became restricted to Muller cells (particularly in the endfeet) and horizontal cell fibers in both rd1/rd1 and +/+. Doublets of alpha GST and mu-GST were detected by Western blot analysis. Densitometry of these bands indicated steady reduction of alpha-GST content in rd1/rd1 retina starting from the second postnatal week. When alpha-GST and mu-GST were added exogenously to rd1/rd1 explants, photoreceptor rescue was produced that was more prominent in PN2 than in PN7 explants and more effective by alpha-GST than mu-GST. We propose that alpha-GST neuroprotection is mediated by reduction of tissue oxidative stress. PMID- 15749347 TI - Major sex differences in non-genomic estrogen actions on intracellular signaling in mouse brain in vivo. AB - Rapid effects of estrogen have now been identified throughout the brain but the extent to which these actions may be different in males and females is unknown. Previous work has shown that estrogen rapidly phosphorylates Ser133 of cAMP responsive element binding protein (CREB) through a non-genomic mechanism. Using this indicator, we have examined here whether non-genomic estrogen actions occur in a sexually dimorphic manner within the adult brain. Male and female mice were gonadectomized and 3 weeks later treated with 17-beta-estradiol or vehicle for 1 h prior to perfusion fixation and subsequent CREB and phosphorylated CREB (pCREB) immunostaining of brain sections. The numbers of cells expressing CREB immunoreactivity were not altered by estrogen treatment or different in males and females in any of the brain regions examined. However, estrogen treatment significantly (P<0.05) increased pCREB-immunoreactive cell numbers in the medial preoptic area, ventrolateral division of the ventromedial nucleus, medial septum and CA1 region of the hippocampus of female mice. In contrast, estrogen increased pCREB levels in the medial septum and CA1 but not in the preoptic area or ventromedial nucleus of male mice. To evaluate the extent to which non-genomic estrogen actions may be sexually differentiated within a single neuronal phenotype, dual labeling immunocytochemistry was undertaken to evaluate the gonadotropin-releasing hormone (GnRH) neuronal phenotype. Estrogen significantly (P<0.05) increased the numbers of GnRH neurons expressing pCREB in female but not male mice. Together, these results demonstrate the existence of a marked sex difference in estrogen's non-genomic effects upon brain function in vivo. PMID- 15749348 TI - The corticotropin-releasing factor (CRF)-system and monoaminergic afferents in the central amygdala: investigations in different mouse strains and comparison with the rat. AB - Corticotropin-releasing-factor (CRF) containing systems and monoaminergic afferents of the central amygdaloid nucleus (Ce) are crucial players in central nervous stress responses. For functional analyses of specific roles of these systems, numerous mouse models have been generated which lack or overexpress individual signal transduction components. Since data concerning system morphologies in murine brain are rarely available, mouse studies are usually designed and interpreted based on previous findings in rats, although interspecies differences are frequent. In the present study, in situ hybridization for CRF mRNA and correlative immunocytochemistry for CRF and monoaminergic afferents revealed numerous CRF mRNA-reactive neurons in the lateral Ce subnucleus (CeL) codistributed with dense dopaminergic fiber plexus in mice as has been demonstrated in rats. However, while in rats the lateral capsular Ce (CeLc) displays only scarce CRF immunoreactive (CRF-ir) innervation, particularly dense CRF-ir fiber plexus were observed in the CeLc in mice, with differences in labeling densities between different strains. CRF-ir terminal fibers overlap with the moderate serotonergic innervation of this subnucleus in mice. Additionally, CRF mRNA-reactive neurons were found immediately dorsal to the amygdala in the region of the interstitial nucleus of the posterior limb of the anterior commissure/amygdalostriatal transition area in both species. In mice, this region displayed dense CRF-ir fiber plexus, with variations between the strains. The results indicate that in mice and rats dopaminergic afferents represent the primary monoaminergic input to the CRF neurons in the CeL. In mice only, CRF-ir afferents provide dense innervation of CeLc neurons. Since the CeLc lacks dopaminergic input in both species but possesses moderate serotonergic afferents, CRF/serotonin interactions may occur selectively in mouse CeLc. The observed interspecies and interstrain differences in CRF input and CRF/monoaminergic interactions may influence the interpretation of findings concerning Ce functions in stress and fear in mouse models. PMID- 15749349 TI - Developmental changes in the spatio-temporal pattern of respiratory neuron activity in the medulla of late fetal rat. AB - We investigated how the spatio-temporal pattern of respiratory neuron network activity in the ventral medulla changes during the late fetal period of rat. Brainstem-spinal cord preparations isolated from rat fetuses on embryonic days 17 21 (E17-E21) were stained with a voltage-sensitive dye for optical image analysis of neuronal activity of the ventral medulla. The spatio-temporal pattern of respiratory neuron activity in the preparation from E20 to E21 was basically identical to that of neonatal rat; pre-inspiratory activity in a limited region of the rostral ventrolateral medulla, the para-facial region, preceded by several hundred milliseconds the onset of inspiratory activity in the more caudal ventrolateral medulla, the pre-Botzinger complex level. In contrast, in E17-E18 specimens, pre-inspiratory activity could not be detected in the rostral medulla at the level of the facial nucleus. Neuronal activity appeared to begin at the pre-Botzinger complex level shortly before onset of the inspiratory burst. Strong activity then developed in the facial nucleus and peaked in the post-inspiratory phase. The transition of these patterns of respiratory activity occurred at E19. We conclude that the changes in the spatio-temporal pattern of neuronal activity reflect developmental changes in the cellular elements underlying rhythm generation in the fetal respiratory neuron network. We suggest that the pre inspiratory neuron network of the para-facial region in the rostral ventrolateral medulla functions as the rhythm generator after E19/20. PMID- 15749350 TI - Fatal embolism of the anterior spinal artery after local cervical analgetic infiltration. AB - A 66-year-old man received a local infiltration of cortisone and xylocain in the left paravertebral region between the fifth and sixth cervical vertebrae. Respiratory failure occurred 2.5 h later with subsequent successful resuscitation. During the following 2 months of assisted ventilation up to the patients death, tetraplegia with concurrent full consciousness throughout was observed. Magnetic resonance imaging confirmed an ischaemic lesion of the upper anterior cervical myelon. The forensic autopsy revealed pneumonia as cause of death. Neuropathology confirmed anterior infarction at the C2/C3 level of the cervical myelon, with obstruction of the anterior spinal artery by an epithelialised fibrocartilaginous embolus. The autoptic findings suggest that this embolus stems from the site of previous analgetic infiltration, with accidental laceration of an intervertebral disc and subsequent transportation of discous material via injection canula into an artery. PMID- 15749352 TI - Preliminary study of post mortem identification using lip prints. AB - Identification using lip prints was first performed in the 1950s and was the subject of much research in the 1960s and 70s, leading to the acceptance of this technique as evidence in the criminal justice system. Previous research has focused on identifying lip print types or on methods of obtaining hidden lip prints left at the crime scene. The present study aimed to clarify characteristics of lip prints from cadavers with various causes of death (including drowning and hanging) and to determine the effects of fixation on post mortem changes in lip impressions. PMID- 15749353 TI - Factors and circumstances influencing the development of hemorrhages in livor mortis. AB - Petechial hemorrhages or ecchymoses in the skin of the face and/or in eyelids and/or conjunctivae are one important feature in postmortem diagnosis of lethal strangulation. On the other hand, petechial bleedings can occur in various causes of death, especially in cases of neck or thoracic compression, they can occur in acute cardiac failure, as a result of blood or skin diseases or as a postmortem phenomenon. The focus of this investigation (retrospective study of 279 corpses, found initially in a prone position or some other face down position) was to analyse the frequency of postmortem (hypostatic) hemorrhages and factors which may influence their development. Petechial hemorrhages in livor mortis in the skin of the trunk and extremities were found in 110 cases (39%). The frequency ranged from 41% in the side position and 44% in the kneeling position to 50% in the prone position. Increasing intensity of livor mortis resulted in an increasing frequency of hemorrhages, up to 59%. In cases with a body-mass-index (BMI) of more than 26 the frequency of hemorrhages increased up to 64%. In cases without livor mortis when the corpses were found as well as in cases with complete movement of livor mortis after turning the corpses, no hemorrhages were found. If hypostasis was partly or completely fixed, the rate of hemorrhages increased up to 50%, without additional increase in longer postmortem intervals. Obviously postmortem petechiae develop neither very soon nor days after death, but within a period of several hours after death. PMID- 15749351 TI - Chest deflection tolerance to blunt anterior loading is sensitive to age but not load distribution. AB - Ninety-three human cadaver tests are used in the development of thoracic injury risk functions with consideration of age and restraint condition. Linear logistic regression models are developed with the set of potential predictors including the maximum chest deflection, the age of the cadaver at death, gender, and the loading condition on the anterior thorax: blunt hub (41 tests), seat belt (26 tests), air bag (12 tests), and combined belt-and-bag (14 tests). Predicted outcomes were the probability of any rib fractures (onset of injury) and the probability of greater than six rib fractures (severe injury). The analysis shows that the injury risk function was not dependent on the loading condition, but was strongly dependent on age. A significant injury risk model with good ability to discriminate injury from non-injury tests (P < 0.0001, chi-square = 21.49, area under receiver operator characteristic curve (ROC) = 0.867, Kruskal's Gamma = 0.732) is presented using only maximum chest deflection and cadaver age as predictors of injury risk. The 50% risk of any rib fractures is found to occur at 35% chest deflection for a 30-year-old, but at 13% deflection for a 70-year-old. The 50% risk of severe injury is shown to occur at 33% chest deflection for a 70 year-old, but at 43% for a 30-year-old. PMID- 15749354 TI - Epidemiological case survey of medical malpractice in some medical and surgical specialties. AB - According to the common view held by the general public, doctors can never make professional mistakes, as if by virtue of their profession they were bound to be infallible. Moreover, when mistakes do occur in the medical field they are seen in a personal light, being attributed to an individual professional figure who is expected to answer for them in both the penal and the civil field. In this paper, the authors made a retrospective analysis of 37 of all the 725 legal suits filed in some hospitals of the Apulian region (South Italy) during the period between 1991 and 2000, being all those lodged against operators in the neurological, urological, otorhino-laryngoiatric and cardiosurgical fields, recorded in the Archives of the Health Services of Bari, Brindisi, Lecce and Taranto or in those of our Forensic Department following consultations on medical and surgical responsibility. PMID- 15749355 TI - Fatal and non-fatal bilateral delayed carotid artery dissection after manual strangulation. AB - Carotid artery (CA) dissection and/or thrombosis may occur in a number of medical and forensic relevant situations. However, post traumatic carotid artery dissection is considered rare, and often underestimated due to possible delayed presentation. We describe two cases of carotid artery dissection following strangulation with delayed symptoms as well as discuss medical diagnostic problems and their forensic implications. PMID- 15749356 TI - Risk factors of sudden death in the Japanese hot bath in the senior population. AB - A series of experiments were carried out to clarify the cause of death and the risk factors related to sudden death in the Japanese senior population while bathing in a Japanese style "hot bath." The biodynamic changes while bathing were carefully monitored under actual bathing situations occurring in both the winter and summer seasons. We observed double product (DP), total peripheral blood vessel resistance (TPR), cardiac output (CO), and blood vessel compliance (COMP) by measuring blood pressure, heart rate, pulse wave, and electrocardiogram (ECG). The finding of a high level of DP in the elderly suggests that more myocardial oxygen consumption is needed than for young adults, particularly in subjects with arrhythmia. Although the values for TPR and CO changed somewhat during bathing, the changes were considered normal and to be expected. However, more significant and substantial changes were observed during the winter experiment than during the summer experiment, no doubt owing to lower temperature of the bathing room. The value of COMP did not vary significantly between winter and summer subjects. Twelve subjects in the elderly developed ECG changes while bathing such as supraventricular extrasystole or ventricular tachycardia. No clinical significance was found in the biochemical analyses of the blood obtained before and after bathing. In conclusion, some subjects in the elderly showed risky changes in the above parameters and ECG, factors which may partially explain some of the causes of the many reported cases of lapse of consciousness and unexpected sudden death in the elderly while bathing especially in the winter season. Cold climate, hot water immersion, and hydrostatic pressure may affect their physiological compensation along with existing of coronary stenosis or weakness of respiratory function as a normal consequence of advanced age. PMID- 15749357 TI - Carbon monoxide-related deaths in a metropolitan county in the USA: an 11-year study. AB - Carbon monoxide (CO) poisoning as a cause of death is well documented in industrialized countries. The objective of this study was to compare demographic data in deaths due to accidents (in fires) and suicides in the same population between 1988 and 1998. Furthermore, the potential effect of a community wide education effort regarding safety in the home was assessed. Postmortem reports were reviewed for all deaths examined at the Office of the Cuyahoga County Coroner in Cleveland, OH, USA. During the study period, there were 209 accidental deaths due to fires in the home (6.5% of all accidents in the home) and 182 CO deaths by suicide (9.8% of all suicides). Demographic characteristics of the two groups differed: while males represented the majority of cases in both groups (55% of accidents, 70% suicides), race specific death rates were higher for whites than blacks (18/100,000 white, 3/100,000 black) in suicides compared with 29/100,000 deaths for blacks and 11/100,000 for whites in accidental cases. Fire deaths were prevalent in the young (0-9 years) and old (>60) whereas in the suicide group the age specific death rate was highest for those over 70 years. The majority of fire deaths occurred in the city of Cleveland but suicides were prevalent in the suburbs. More fire deaths occurred in December than any other month whereas more suicides occurred in April. In 1992, there was a community wide effort to provide free smoke detectors to residents in Cleveland. In 1992, there were 4.2/100,000 fire deaths in the city. This decreased to 0.6/100,000 in 1996, increased to 1.2/100,000 in 1997 followed by a decrease to 0.8/100,000 in 1998. This suggested that the program may have aided in decreasing these types of deaths. Deaths due to fires in the suburbs were <1/100,000 throughout the study period. PMID- 15749358 TI - A fatal poisoning caused by methomyl and nicotine. AB - A 35-year-old male was found lying in a prone position in his room. He was in cardiopulmonary arrest on arrival to hospital and was pronounced dead. There was no attempt at resuscitation. No miosis was observed on admission. At post-mortem his stomach contained 170 g greenish liquid with a small amount of shredded tobacco leaves. The serum cholinesterase activities were 47-90 IU (normal range for male: 200-440 IU). GC and GC-MS analyses showed nicotine (21.8 mg), methomyl (304 mg), and triazolam (1.69 mg) in his stomach. He had consumed tobacco leaves, Lannate containing water soluble methomyl (45%), and Halcion tablets containing 0.25 mg triazolam. Methomyl concentrations in blood were 3-8 ng/ml. Substantial amounts of methomyl (2260-2680 ng/ml) were detected in cerebrospinal fluid and vitreous humor. Nicotine concentrations in blood ranged from 222 to 733 ng/ml. A small amount of triazolam was detected only in bile (176 ng/ml) and liver (23 ng/g). The cause of death was respiratory paralysis produced by the additive effects of methomyl and nicotine shortly after consumption. PMID- 15749360 TI - Determination of synthesis route of 1-(3,4-methylenedioxyphenyl)-2-propanone (MDP 2-P) based on impurity profiles of MDMA. AB - In our study 1-(3,4-methylenedioxyphenyl)-2-propanone (MDP-2-P or PMK) was prepared by two different routes, i.e. by oxidizing isosafrole in an acid medium and by 1-(3,4-methylenedioxyphenyl)-2-nitropropene reduction. The final product MDP-2-P was subjected to GC/MS analysis. The intermediates and reaction by products were identified and the 'route specific' impurities were established. The following impurities are the markers of the greatest importance: 1-(3,4 methylenedioxyphenyl)-1-propanone (compound 10, Table 2), 1-methoxy-1-(3,4 methylenedioxyphenyl)-2-propanone (compound 11, Table 2) and 2,2,4-trimethyl-5 (3,4-methylenedioxyphenyl)-[1,3]dioxolane (compound 13, Table 2) (the 'oxidising isosafrole route') and N-cyclohexylacetamide (compound 3, Table 1), 3-methyl-6,7 methylenedioxyisoquinoline-1,4-dione (compound 15, Table 1) (the 'MDP-2 nitropropene reduction route'). Subsequently, MDMA was prepared by reductive amination of MDP-2-P using NaBH4 as reducing agent (so-called 'cool method'). Impurities were extracted with n-heptane under alkaline conditions. The impurity profiles were obtained by means of GC/MS, some reaction by-products were identified by means of the EI mass spectra including low energy EI mass spectra and 'route specific' impurities were established. 4-Methyl-5-(3,4 methylenedioxyphenyl)-[1,3]dioxolan-2-one (compound 22, Table 2), N-methyl-2 methoxy-1-methyl-2-(3,4-methylenedioxyphenyl)-ethaneamine (compound 18, Table 2), 3-methyl-6,7-methylenedioxyisoquinoline-1,4-dione (compound 15, Table 1) and N cyclohexyloacetamide (compound 3, Table 1) were found to be the synthesis markers of greatest importance. PMID- 15749359 TI - Urinary endogenous concentrations of GHB and its isomers in healthy humans and diabetics. AB - Urinary endogenous concentrations of gamma-hydroxybutyric acid (GHB), alpha hydroxybutyric acid (AHB) and beta-hydroxybutyric acid (BHB) have been investigated for both healthy humans and diabetics by using a newly optimized GC MS procedure. The endogenous concentrations in healthy volunteers' urine ranged 0.16-2.14 microg/ml for GHB, 0.10-2.68 microg/ml for AHB and 8.51-34.7 microg/ml for BHB. In diabetics, the concentrations ranged 0.17-3.03 microg/ml for GHB, 0.14-124 microg/ml for AHB and 4.94-4520 microg/ml for BHB. Although notably elevated BHB and AHB concentrations were observed for severely uncontrolled diabetics, their GHB concentrations ranged within or near the range seen in healthy humans. The results of this study confirm the previously suggested 10 microg/ml cutoff concentration of urinary GHB to distinguish exogenous GHB, even for uncontrolled diabetic patients suffering severe ketoacidosis. PMID- 15749361 TI - Employing glass refractive index measurement (GRIM) in fiber analysis: a simple method for evaluating the crystallinity of acrylics. AB - The refractive index (RI) of 40 colorless acrylic fiber samples was determined by the immersion oil and Mettler hot stage method, implemented in the glass refractive index measurement (GRIM) instrument by Foster and Freeman. Low standard deviations for nearly all the samples were obtained and differentiation of fibers even with the same trade name was possible just on the basis of RI. Wide angle X-ray scattering (WAXS) and differential scanning calorimetry (DSC) were employed to establish a linear correlation between RI and the degree of crystallinity. Insights on the structural order of this group of fibers is then obtainable by a quick and easy technique, adding value to casework comparisons. PMID- 15749362 TI - Calculating the exclusion probability and paternity index for X-chromosomal loci in the presence of substructure. AB - There has been recent interest in the use of X-chromosomal loci for forensic and relatedness testing casework, with many authors developing new X-linked short tandem repeat (STR) loci suitable for forensic use. Here we present formulae for two key quantities in paternity testing, the average probability of exclusion and the paternity index, which are suitable for X-chromosomal loci in the presence of population substructure. PMID- 15749363 TI - Apnea, glial apoptosis and neuronal plasticity in the arousal pathway of victims of SIDS. AB - Of 27,000 infants whose sleep-wake characteristics were studied under the age of 6 months, 38 died unexpectedly 2-12 weeks after the sleep recording in a pediatric sleep laboratory. Of these infants, 26 died of sudden infant death syndrome (SIDS), and 12 of definitely identified causes. The frequency and duration of sleep apneas were analysed. Sleep recordings and brainstem histopathology were studied to elucidate the possible relationship between sleep apnea and neuropathological changes within the arousal system. Immunohistochemical analyses were conducted using tryptophan hydroxylase (TrypH), a serotonin synthesizing enzyme, and growth-associated phosphoprotein 43 (GAP43), a marker of synaptic plasticity. The terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method was used for apoptosis. The pathological and physiological data were correlated for each infant. In the SIDS victims, statistically significant positive correlations were seen between the number of TrypH-positive neurons in the dorsal raphe nucleus of the midbrain and the duration of central apneas (p = 0.03), between the number of TUNEL-positive glial cells in the pedunculopontine tegmental nucleus (PPTN) and the average number of spines in GAP43-positive neurons in the PPTN (p = 0.04). These findings in the dorsal raphe nucleus of the midbrain and PPTN, that play important roles in the arousal pathway suggest a possible link between changes in arousal and SIDS. PMID- 15749364 TI - Development of a harmonized method for the profiling of amphetamines. I. Synthesis of standards and compilation of analytical data. AB - Reference material was synthesised for 21 substances that are frequently present as synthetic impurities, i.e. by-products, in illicitly produced amphetamine. Each of these substances is a typical by-product for at least one of the three approaches most often used to synthesise amphetamine, namely, the Leuckart, the reductive amination of benzyl methyl ketone, and the nitrostyrene routes. A large body of data on the substances was recorded, including the following: mass spectra, ultraviolet spectra, Fourier transform infrared spectra, infrared spectra in gas phase, and 1H NMR and 13C NMR spectra. PMID- 15749365 TI - Development of a harmonized method for the profiling of amphetamines. II. Stability of impurities in organic solvents. AB - The present study focused on the stability of 22 amphetamine impurities dissolved in six organic solvents: isooctane, toluene, ethanol, dichloromethane, ethyl acetate, and diethyl ether. The aim was to find the most inert, and thereby most suitable, solvent for amphetamine profiling. Mixtures of the impurities were prepared in the different solvents, and changes in the concentrations of the individual compounds over-time were monitored by gas chromatographic analysis after 0, 4, 12, 24, 48, and 96 h. Isooctane and toluene provided the most inert conditions, although, a few of the impurities were insufficiently stable in these two solvents. The present experiments were performed as a part of the development of a harmonized method for profiling of amphetamine. The results can be used to support the choice of organic solvents for sample preparation. They also provide information about the stability of the impurities that are found in profiles of illicit amphetamine. This is essential due to the fact, that unstable compounds can have a negative influence on the comparison of profiles. PMID- 15749366 TI - A fatal case of pure ethanol ingestion. AB - An adult male was found dead in a car with two empty bottles (500 ml x 2) labeled dehydrated ethanol (>99.5%, v/v). At autopsy, extensive pancreatic necrosis with severe hemorrhage was observed. High concentrations of ethanol were detected in blood (8.14 mg/ml), urine (8.12 mg/ml) and tissue specimens. The cause of death was determined to be an acute alcohol intoxication caused by ingesting approximately 1l dehydrated ethanol. PMID- 15749368 TI - Fatal colchicine poisoning by accidental ingestion of meadow saffron-case report. AB - A 62-year-old male died of colchicine poisoning after accidental ingestion of Colchicum autumnale (meadow saffron). He ate a salad of plant with green leaves regarded as wild garlic (Allium ursinum). A few hours later he developed symptoms of gastroenteritis and was admitted to hospital. In spite of gastric lavage, activated charcoal and supportive measures, multi-organ system failure developed over the next two days. Laboratory analysis showed highly elevated blood concentrations of hepatic enzymes, creatine kinase, lactate dehydrogenase and blood urea nitrogen, as well as leukocytopenia and thrombocytopenia. Mechanical ventilation, dopamine, noradrenaline, crystalloid solutions and fresh frozen plasma were applied but despite treatment the patient died five days after the ingestion. Post-mortem examination revealed hepatic centrilobular necrosis, nephrotoxic acute tubular necrosis, petechial bleeding in fatty tissue, blunt and shortened intestinal villi and cerebral toxic edema. Botanical identification of incriminated plant gave Colchicum autumnale which confirmed colchicine poisoning. Although the accidental ingestion of Colchicum autumnale is rare and to our knowledge only five such cases have been described in detail, this is the second fatal case in Croatia described in the last 3 years. PMID- 15749367 TI - A fatality involving metaxalone. AB - A case is presented of a 54-year-old white female found dead in a secured apartment. Postmortem toxicologic analysis of the heart blood identified acetaminophen (97 mg/L), citalopram (0.4 mg/L), gabapentin (24 mg/L) and metaxalone (21 mg/L). The metaxalone concentration is within the range of previously reported fatalities involving metaxalone. The medical examiner ruled that the cause of death was metaxalone and gabapentin intoxication and the manner of death was suicide. PMID- 15749369 TI - Human Y-specific STR haplotypes in the Western Croatian population sample. AB - Nine Y-chromosome STR loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385 and YCAII) were analysed in a sample of 101 unrelated males from Croatia. Allelic frequencies and gene diversities for each Y-STR locus and haplotype diversity were determined. Ninety-one different haplotypes were obtained from 101 unrelated males and 84 haplotypes were unique. Three most common haplotypes were shared by 3% of the sample, one of them was not found in the online Y-STR Haplotype Reference Database (http://www.ystr.org/). PMID- 15749370 TI - Genetic data on 15 STR loci in the ethnic group of Polish Tatars residing in the area of Podlasie (Northeastern Poland). AB - Population genetic data for the 15 STRs included in the AmpFlSTR Identifiler kit were obtained from a sample of 168 unrelated individuals of the ethnic group of Polish Tatars residing in the Northeastern Poland. Significant differences revealed in relation to the autochthonous Poles by using R x C test as well as F(ST) and F(IS) estimates suggest that the ethnic group of Polish Tatars is a distinct genetic community. PMID- 15749371 TI - Haplotype diversity in mitochondrial DNA hypervariable region I, II and III in northeast China Han. AB - Sequence polymorphism of the mitochondrial DNA (mtDNA) control region, hypervariable regions HVR I, HVR II and HVR III, from 51 unrelated China Han (Yan Bian area) were determined by PCR amplification and cycle sequencing. PMID- 15749372 TI - Mitochondrial DNA control region sequence polymorphism in four indigenous tribes of Chotanagpur plateau, India. AB - The analysis of genetic variation, in the nucleotide sequences of mitochondrial DNA, provides unique information in tracing of maternal lineage, determination of population diversity, pharmacogenomics and human identification. This study characterizes the HVR-I and II sequence polymorphism in 80 tribal individuals, belonging to the Austro-Asiatic linguistic family of Chotanagpur plateau, India. A total of 115 polymorphic sites were observed in the sequenced regions and 77 unique haplotypes could be identified. PMID- 15749373 TI - Identification of more sequence variations in the D8S1179 locus. AB - Routine STR-typing of 10,293 buccal swabs using different multiplex kits presented discordant D8S1179 profiles in four cases. Sequencing analysis identified a G-to-A transition upstream to the repeat, and an A-to-T transversion and a G-to-A transition downstream to the repeat. In the fourth case a four-base pair deletion downstream resulted in altered genotypes using different primer pairs. Current searching algorithms of the German DNA database are not capable of matching profiles that are divergent in only one STR-locus. Thus, to accommodate matching requirements and to avoid errors in individual genetic characterization for D8S1179, as described here, it is suggested that alternative primer pairs be used for routine genotyping as a matter of course. PMID- 15749374 TI - Allele frequencies for 70 autosomal SNP loci with U.S. Caucasian, African American, and Hispanic samples. AB - 189 samples from 3 different U.S. sample groups Caucasian (74), African American (71) and Hispanic (44) were typed for 70 autosomal genetic markers. These 70 markers are bi-allelic (C/T) short nucleotide polymorphisms (SNPs). For each sample, the 70 SNP markers were typed in 11 unique 6-plexes and a single 4-plex PCR. A total of 10 of the 210 tests (70 loci x 3 populations) for Hardy-Weinberg equilibrium indicated a statistically significant result. In order to evaluate the minimum number of SNP loci needed to distinguish all 189 samples from one another, we ranked the loci according to their levels of observed heterozygosity and p-values obtained upon testing for Hardy-Weinberg equilibrium. The top 12 loci according to these ranking criteria were tabulated along with the number of unique genotypes observed when combining subsequent SNP markers. The 12 selected SNPs possessed an observed heterozygosity of >0.45 in all three populations examined and thus would be expected to exhibit more differences between samples. All of the 189 samples in this study were individualized with a subset of 12 SNP loci. However, it is likely that the addition of more than 12 SNP loci will be required to resolve larger sets of unrelated individuals from one another. By way of comparison, in these same 189 individuals all but one pair is resolved from one another with three of the traditional short tandem repeat (STR) loci possessing the highest heterozygosity values (D2S1338, D18S51, and FGA) run with the Identifiler kit. The final pair of unrelated samples could be resolved with the combination of 4 STR loci: D2S1338, D18S51, FGA, and VWA. PMID- 15749375 TI - Fatal alcohol immersion during the SARS epidemic in Taiwan. PMID- 15749376 TI - Interpretation of postmortem alcohol concentrations. PMID- 15749377 TI - Commentary on: Mark Benecke and Larry Barksdale, Distinction of bloodstain patterns from fly artifacts. Forensic Science International. 137(2003) 152-159. PMID- 15749378 TI - S-Nitrosothiols: cellular formation and transport. AB - This review will focus on the transport and intracellular formation of S nitrosothiols in cell culture models. The major points made in this article are: (1) S-Nitrosothiols are actively metabolized by cells. (2) S-Nitrosothiols affect cells in ways distinctly different from those of nitric oxide and can act through mechanisms that do not involve the intermediacy of nitric oxide. (3) Some S nitrosothiols (S-nitrosocysteine, S-nitrosohomocysteine) can be taken up into cells via amino acid transport system L, whereas others (S-nitrosoglutathione, S nitroso-N-acetylpenicillamine) are not directly transported, but require the presence of cysteine and/or cystine before the nitroso functional group is transported. (4) Proteomic detection of intracellular S-nitrosothiols is currently possible only if cells are loaded with high levels of S-nitrosothiols, and methodological advances are required in order to examine the S-nitrosated proteome after exposure of cells to physiological levels of nitric oxide. PMID- 15749379 TI - Osmium tetroxide, used in the treatment of arthritic joints, is a fast mimic of superoxide dismutase. AB - Aqueous solutions of osmium tetroxide (OsO4) have been injected into arthritic knees for the past 45 years to chemically destroy diseased tissue, in a procedure termed "chemical synovectomy." Arthritis is an inflammatory disease. The primary inflammatory chemical species are the superoxide anion radical (O2.-) and nitric oxide (.NO), which combine to form the peroxynitrite anion (ONOO-). Here we show that OsO4 does not react with ONOO- but very efficiently catalyzes the dismutation of O2.- to O2 and H2O2. Using the pulse-radiolysis technique, the catalytic rate constant has been determined to be (1.43+/-0.04) x 10(9) M-1 s-1, independent of the pH in the 5.1-8.7 range. This value is about half that for the natural Cu,Zn-superoxide dismutase (Cu,Zn-SOD). Per unit mass, OsO4 is about 60 times more active than Cu,Zn-SOD. The catalytically active couple is OsVIII/OsVII, OsVIII oxidizing O2.- to O2 with a bimolecular rate constant of k=(2.6+/-0.1)x10(9) M-1 s-1 and OsVII reducing it to H2O2 with a bimolecular rate constant of (1.0+/-0.1)x10(9) M-1 s-1. Although lower valent osmium species are intrinsically poor catalysts, they are activated through oxidation by O2.- to the catalytic OsVIII/OsVII redox couple. The OsVIII/OsVII catalyst is stable to biochemicals other than proteins and peptides comprising histidine, cysteine, and dithiols. PMID- 15749380 TI - Central role of PKCdelta in glycoxidation-dependent apoptosis of human neurons. AB - Accumulation of advanced glycation end products (AGEs) induces alterations in the intracellular redox balance, leading cells to functional injury. Current literature reports that intracellular signaling triggered by the interaction of AGEs with their specific receptors RAGEs depends on the cell type and the state of activation/stress. In this work, NT2 human neurons were exposed for 48 h to glycated fetal serum containing 750-3000 pmol/ml pentosidine; the treatment induced an increase in apoptosis rate linear with AGE concentration up to 1500 pmol/ml, but necrotic death was elicited with the highest AGE amount employed (3000 pmol/ml pentosidine). Pentosidine at 1500 pmol/ml, which was the concentration responsible for the highest apoptotic effect (40% of apoptotic neurons), was able to determine early generation of intracellular reactive oxygen species and increase in RAGE levels. Under these conditions, protein kinase C (PKC) delta activity was increased approximately 2-fold, and DNA binding activity of redox-sensitive transcription factor activator protein-1 (AP-1) was enhanced 2.5-fold. A relationship among oxidative stress, PKCdelta activity, AP-1 activation, and apoptosis was demonstrated by pretreating neurons with 500 muM vitamin E, with 20 mug/ml Ginkgo biloba extract, or with 3 muM Rottlerin, inhibitor of PKCdelta; these pretreatments were able to protect neurons from the glycoxidation-dependent effects. PMID- 15749381 TI - Studies in humans using deuterium-labeled alpha- and gamma-tocopherols demonstrate faster plasma gamma-tocopherol disappearance and greater gamma metabolite production. AB - We hypothesized that human plasma alpha- and gamma-tocopherol concentrations reflect differences in their kinetics, especially influenced by gamma-tocopherol metabolism. Vitamin E kinetics were evaluated in humans (n=14) using approximately 50 mg each of an equimolar ratio of d6-alpha- and d2-gamma tocopheryl acetates administered orally. Mass spectrometry was used to measure deuterated plasma tocopherols, as well as plasma and urinary vitamin E metabolites, alpha- and gamma-carboxyethylhydroxychromans (CEHCs). Plasma d2 gamma-tocopherol fractional disappearance rates (FDR; 1.39+/-0.44 pools/day, mean+/-SD) were more than three times greater than those of d6-alpha-tocopherol (0.33+/-0.11, p<0.001). The d2-gamma-tocopherol half-life was 13+/-4 h compared with 57+/-19 for d6-alpha-tocopherol. Whereas neither plasma nor urinary d6-alpha CEHC was detectable (limit of detection 1 nmol/L), gamma-CEHC (labeled plus unlabeled) increased from 129+/-20 to 258+/-40 nmol/L by 12 h and returned to baseline by 48 h; at 12 h d2-gamma-CEHC represented 54+/-4% of plasma gamma-CEHC. Women compared with men had a greater d2-gamma-tocopherol FDR (p<0.004) and a greater maximal plasma d2-gamma-CEHC concentration (p<0.02) and CEHC FDR (p<0.007), as well as excreting four times as much d2-gamma-CEHC (p<0.04) in urine. Thus, gamma-tocopherol is rapidly metabolized to gamma-CEHC, and to a greater degree in women than in men, whereas alpha-tocopherol is maintained in the plasma and little is metabolized to alpha-CEHC. PMID- 15749382 TI - The biosynthesis of ascorbate protects isolated rat hepatocytes from cumene hydroperoxide-mediated oxidative stress. AB - Most animals synthesize ascorbate. It is an essential enzymatic cofactor for the synthesis of a variety of biological molecules and also a powerful antioxidant. There is, however, little direct evidence supporting an antioxidant role for endogenously produced ascorbate. Recently, we demonstrated that incubation of rat hepatocytes with 1-bromoheptane or phorone simultaneously depleted glutathione (GSH) and triggered rapid ascorbate synthesis. The present study investigates the hypothesis that endogenous ascorbate synthesis can confer protection against oxidative stress. Rat and guinea pig hepatocytes were depleted of GSH with 1 bromoheptane and subsequently treated with the oxidative stressor cumene hydroperoxide (CHP) in the presence or absence of the ascorbate synthesis inhibitor sorbinil. In rat hepatocytes, ascorbate content increased linearly (from 15.1 to 35.8 nmol/10(6) cells) over a 105-min incubation. Prior depletion of GSH increased CHP-induced cellular reactive oxygen species (ROS) production, lipid peroxidation, and cell death in rat and guinea pig hepatocytes. Inhibiting ascorbate synthesis, however, further elevated ROS production (2-fold), lipid peroxidation (1.5-fold), and cell death (2-fold) in rat hepatocytes only. This is the first time that endogenous ascorbate synthesis has been shown to decrease cellular susceptibility to oxidative stress. Protection by endogenously produced ascorbate may therefore need to be addressed when extrapolating data to humans from experiments using rodents capable of synthesizing ascorbate. PMID- 15749383 TI - Characterization and application of the biotin-switch assay for the identification of S-nitrosated proteins. AB - S-Nitrosation of protein cysteinyl residues has been suggested to be an important nitric oxide-dependent posttranslational modification. The so-called biotin switch method has been developed to identify S-nitrosated proteins. This method relies on the selective reduction of S-nitrosothiols by ascorbate. In this study we have assessed the ability of ascorbate to reduce S-nitrosothiols and show that ascorbate is a very inefficient reducing agent. We show that higher concentrations of ascorbate and longer incubation times can significantly improve immunological detection of S-nitrosothiols. We have compared immunological detection of S-nitrosothiols with the level of intracellular S-nitrosothiols measured by tri-iodide chemiluminescence and show that the biotin-switch method is capable of detecting only high (nmol/mg protein) levels of intracellular S nitrosothiols obtained after exposing cells to S-nitrosocysteine, but not the low levels observed during physiological nitric oxide formation. Preliminary proteomic analysis of protein S-nitrosothiols has identified elongation factor 2, heat shock protein 90 beta, and a 65-kDa macrophage protein homologous to human L plastin as major nitrosation targets at high intracellular nitrosation levels in the murine macrophage-derived RAW 264.7 cell line. While the biotin-switch method may be a useful tool to aid in the positive identification of protein S nitrosothiols, it cannot match the sensitivity of chemiluminescence-based methods and its use in proteomic studies likely suffers from selective detection of more easily reducible S-nitrosothiols. PMID- 15749384 TI - Effects of oxidative stress on adiponectin secretion and lactate production in 3T3-L1 adipocytes. AB - Obesity is an increasing nutritional disorder in developed countries, and oxidative stress has been identified as a key factor in numerous pathologies such as diabetes, inflammation, and atherosclerosis, which are favored by obesity. The objective of the present study was to investigate the effects of oxidative stress in 3T3-L1 adipose cells on two parameters involved in metabolic complications associated with obesity, namely adiponectin secretion and lactate production. Differentiated 3T3-L1 adipose cells were exposed to increasing concentrations of glucose oxidase. 4-Hydroxynonenal (4-HNE), a relevant lipid peroxidation by product which may affect several metabolic processes in making covalent adducts with various molecules; adiponectin secretion; and lactate production were measured in response to glucose oxidase exposure. Results show an inhibition of adiponectin mRNA expression by glucose oxidase and a significant inverse correlation between 4-HNE formation and adiponectin secretion. Furthermore, 4-HNE alone inhibits adiponectin production by 3T3-L1. On the other hand, glucose oxidase and 4-HNE significantly stimulated lactate production by 3T3-L1 adipocytes. These results demonstrate that adipose cells are highly sensitive to oxidative stress, with subsequent decreased adiponectin secretion and increased lactate production, two events involved in the development of insulin resistance. PMID- 15749385 TI - Involvement of UVB-induced reactive oxygen species in TGF-beta biosynthesis and activation in keratinocytes. AB - TGF-beta produced by keratinocytes in response to UVB (290-320 nm) is a potential mediator for effects of acute and chronic solar radiation on skin. This study was designed to determine whether reactive oxygen species (ROS) mediate UVB-induced TGF-beta biosynthesis in keratinocytes and the subsequent activation of the latent TGF-beta complex. UVB irradiation elevated both total (latent plus active) and active TGF-beta in the keratinocyte supernatants, with a greater increase in the active form. UVB irradiation induced up to a 30% increase in ROS, and the ROS were detected up to 90 min after irradiation. NAC and Trolox, cytoplasmic ROS scavengers, abolished the UVB-induced TGF-beta and intracellular ROS, suggesting that UVB-induced ROS are involved in TGF-beta regulation. Inhibitors of NADPH oxidase activity, DPI and apocynin, decreased UVB-induced ROS. The increase in NADPH oxidase activity was mediated by EGFR activation. UVB-induced ROS also activated latent TGF-beta complex by stimulating MMP-2 and -9 activities. In summary, physiological doses of UVB increase intracellular ROS, which upregulate TGF-beta biosynthesis and activation of TGF-beta through increased activity of MMPs. PMID- 15749386 TI - Vitamin C homeostasis in skeletal muscle cells. AB - In skeletal muscle, vitamin C not only enhances carnitine biosynthesis but also protects cells against ROS generation induced by physical exercise. The ability to take up both ascorbic and dehydroascorbic acid from the extracellular environment, together with the ability to recycle the intracellular vitamin, maintains high cellular stores of ascorbate. In this study, we examined vitamin C transport and recycling, by using the mouse C2C12 and rat L6C5 muscle cell lines, which exhibit different sensitivity to oxidative stress and GSH metabolism. We found that: (1) both cell lines express SVCT2, whereas SVCT1 is expressed at very low levels only in proliferating L6C5 cells; furthermore L6C5 myoblasts are more efficient in ascorbic acid transport than C2C12 myoblasts; (2) C2C12 cells are more efficient in dehydroascorbic acid transport and ascorbyl free radical/dehydroascorbic acid reduction; (3) differentiation is paralleled by decreased ascorbic acid and dehydroascorbic acid transport and reduction and increased ascorbyl free radical reduction; (4) differentiated cells are more responsive to oxidative stress induced by glutathione depletion; indeed, myotubes showed increased SVCT2 expression and thioredoxin reductase-mediated dehydroascorbic acid reduction. From our data, SVCT2 and NADPH-thioredoxin dependent DHA reduction appears to belong to an inducible system activated in response to oxidative stress. PMID- 15749387 TI - Hydroxytyrosol, a natural antioxidant from olive oil, prevents protein damage induced by long-wave ultraviolet radiation in melanoma cells. AB - Previous studies showed that long-wave ultraviolet (UVA) radiation induces severe skin damage through the generation of reactive oxygen species and the depletion of endogenous antioxidant systems. Recent results from our laboratory indicate a dramatic increase of both lipid peroxidation products (TBARS) and abnormal L isoaspartyl residues, marker of protein damage, in UVA-irradiated human melanoma cells. In this study, the effects of hydroxytyrosol (DOPET), the major antioxidant compound present in olive oil, on UVA-induced cell damages, have been investigated, using a human melanoma cell line (M14) as a model system. In UVA irradiated M14 cells, a protective effect of DOPET in preventing the uprise of typical markers of oxidative stress, such as TBARS and 2'7'-dichlorofluorescein (DCF) fluorescence intensity, was observed. In addition, DOPET prevents the increase of altered L-isoAsp residues induced by UVA irradiation. These protective effects are dose dependent, reaching the maximum at 400 microM DOPET. At higher concentrations, DOPET causes an arrest of M14 cell proliferation and acts as a proapoptotic stimulus by activating caspase-3 activity. In the investigated model system, DOPET is quantitatively converted into its methylated derivative, endowed with a radical scavenging ability comparable to that of its parent compound. These findings are in line with the hypothesis that the oxidative stress plays a major role in mediating the UVA-induced protein damage. Results suggest that DOPET may exerts differential effects on melanoma cells according to the dose employed and this must always be taken into account when olive oil-derived large consumer products, including cosmetics and functional foods, are employed. PMID- 15749388 TI - In vitro photodynamic effects of phthalocyaninatosilicon covalently linked to 2,2,6,6-tetramethyl-1-piperidinyloxy radicals on cancer cells. AB - In this paper, we have investigated the ability to sensitize the phototoxicity toward HeLa cells in vitro, of tetra-tert-butylphthalocyaninatosilicon (SiPc) covalently linked to one or two 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) radicals (R1c or R2c), which are shown as photosensitizers efficiently producing singlet oxygen (1Delta(g)). Addition of R1c or R2c encapsulated in liposomes to cultures, followed by irradiation with a 680-nm dye laser, resulted in a highly significant phototoxicity toward HeLa cells, in contrast to negligible phototoxicity observed with (dihydroxy)SiPc (R0). EPR measurements indicate that R1c and R2c exist in some degree as nitroxide radicals even in HeLa cells. Electronic absorption spectra indicate that the degree of aggregation increases in the order R2c nSMase --> ceramide --> free radical --> cell death" pathway in primary cultures of fetal rat cortical neurons. We also provided experimental evidence showing that S-nitrosoglutathione (GSNO), a potent endogenous antioxidant derived from the interaction between nitric oxide (NO) and glutathione, caused dose-dependent protective effects against Abeta/ceramide neurotoxicity via inhibition of caspase activation and production of reactive oxygen species (ROS). This GSNO-mediated neuroprotection appeared to involve activation of cGMP-dependent protein kinase (PKG), phosphatidylinositol 3-kinase (PI3K), and extracellular signal-regulated kinase (ERK). Activation of the cGMP/PKG pathway induced expression of thioredoxin and Bcl-2 that were beneficial to cortical neurons in antagonizing Abeta/ceramide toxicity. Consistently, exogenous application of thioredoxin exerted remarkable neuroprotective efficacy in our experimental paradigm. Results derived from the present study establish a neuroprotective role of GSNO, an endogenous NO carrier, against Abeta toxicity via multiple signaling pathways. PMID- 15749395 TI - Using simulation to teach resuscitation: an important patient safety tool. AB - Learning resuscitation scenarios using high-fidelity simulation can have a positive impact on real-life learning for health care professionals. Simulation sessions encourage knowledge acquisition and critical thinking for nursing students, experienced nurses, nurse anesthetists, physicians, and respiratory therapists. A novice to expert theoretical perspective can be accomplished without anxiety over scenario outcome. Results of educational endeavors support simulation use, but continued research is needed to substantiate learner outcome and technologic value. PMID- 15749393 TI - Identification of the myoglobin tyrosyl radical by immuno-spin trapping and its dimerization. AB - 5,5-Dimethyl-1-pyrroline N-oxide (DMPO) spin trapping in conjunction with antibodies specific for the DMPO nitrone epitope was used on hydrogen peroxide treated sperm whale and horse heart myoglobins to determine the site of protein nitrone adduct formation. The present study demonstrates that the sperm whale myoglobin tyrosyl radical, formed by hydrogen peroxide-dependent self peroxidation, can either react with another tyrosyl radical, resulting in a dityrosine cross-linkage, or react with the spin trap DMPO to form a diamagnetic nitrone adduct. The reaction of sperm whale myoglobin with equimolar hydrogen peroxide resulted in the formation of a myoglobin dimer detectable by electrophoresis/protein staining. Addition of DMPO resulted in the trapping of the globin radical, which was detected by Western blot. The location of this adduct was demonstrated to be at tyrosine-103 by MS/MS and site-specific mutagenicity. Interestingly, formation of the myoglobin dimer, which is known to be formed primarily by cross-linkage of tyrosine-151, was inhibited by the addition of DMPO. PMID- 15749391 TI - Levels of reactive oxygen species and primary antioxidant enzymes in WI38 versus transformed WI38 cells following bleomcyin treatment. AB - Bleomycin (BLM) is an anticancer drug that generates reactive oxygen species (ROS) after interacting with iron and oxygen. We hypothesized that BLM could cause a different status of oxidative stress in normal versus tumor cells due to possible altered redox status and gene expression in cells following transformation. In this study, the extent of cytotoxicity, levels of ROS, and activities of antioxidant enzymes were compared between normal WI38 cells and SV40-transformed WI38 (VA13) cells following BLM treatment. Basal activities of MnSOD and catalase were lower in VA13 cells and basal ROS levels were higher in VA13 cells. Although BLM caused greater growth inhibition and apoptosis in VA13 cells, it increased ROS levels at an earlier time point in WI38 cells. Moreover, BLM treatment (100 microg/ml) had no effect on the activities of MnSOD, CuZnSOD, and catalase, but increased the activities of glutathione peroxidase (GPX) in WI38 cells after a 48-h treatment and in VA13 cells after a 24- and 48-h treatment. Northern blot analysis indicated that the increase in GPX activities was due to increased transcript levels of GPX1 but not GPX4 in both cells. Our results indicate selective induction of the GPX1 gene by BLM and different redox responses to BLM between WI38 and VA13 cells. PMID- 15749392 TI - Proteomic analysis of 4-hydroxy-2-nonenal-modified proteins in G93A-SOD1 transgenic mice--a model of familial amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is an age-related, fatal motor neuron degenerative disease occurring both sporadically (sALS) and heritably (fALS), with inherited cases accounting for approximately 10% of diagnoses. Although multiple mechanisms likely contribute to the pathogenesis of motor neuron injury in ALS, recent advances suggest that oxidative stress may play a significant role in the amplification, and possibly the initiation, of the disease. Lipid peroxidation is one of the several outcomes of oxidative stress. Since the central nervous system (CNS) is enriched with polyunsaturated fatty acids, it is particularly vulnerable to membrane-associated oxidative stress. Peroxidation of cellular membrane lipids or circulating lipoprotein molecules generates highly reactive aldehydes, among which is 4-hydroxy-2-nonenal (HNE). HNE levels are increased in spinal cord motor neurons of ALS patients, indicating that lipid peroxidation is associated with the motor neuron degeneration in ALS. In the present study, we used a parallel proteomic approach to identify HNE-modified proteins in the spinal cord tissue of a model of fALS, G93A-SOD1 transgenic mice, in comparison to the nontransgenic mice. We found three significantly HNE modified proteins in the spinal cord of G93A-SOD1 transgenic mice: dihydropyrimidinase-related protein 2 (DRP-2), heat-shock protein 70 (Hsp70), and possibly alpha-enolase. These results support the role of oxidative stress as a major mechanism in the pathogenesis of ALS. Structural alteration and activity decline of functional proteins may consistently contribute to the neurodegeneration process in ALS. PMID- 15749396 TI - Implantable cardioverter defibrillator storm: nursing care issues for patients and families. AB - Implantable cardioverter defibrillators (ICDs) are being used for primary and secondary prevention of life-threatening cardiac arrhythmias, and evidence suggests that increased use is likely in the future. ICD storm, the delivery of two or more shocks within 24 hours, occurs in 10% to 20% of patients who have ICDs and can have long-lasting psychological and physical consequences. An understanding of the factors associated with ICD storm, relevant assessment, and patient and family teaching and counseling can help clinicians to better meet the needs of patients who have experienced ICD storm. PMID- 15749397 TI - Pediatric post-resuscitation care. AB - Current literature demonstrates a paucity of information on post-resuscitation care of pediatric clients. This lack of information is somewhat understandable in light of the relatively low incidence of occurrence and the statistically poor outcome. Nurses must be aware, however, of many issues when dealing with pediatric clients and their families after an arrest episode. This article explores key concepts involved with post-resuscitation care, including the outcome of cardiopulmonary resuscitation, immediate post-resuscitation needs, emotional outcomes for the child, and family stress and grief. PMID- 15749398 TI - Family presence during cardiopulmonary resuscitation. AB - A recent phenomenon in emergency and critical care settings is the presence of family members during resuscitation events. It remains controversial in most institutions, but evidence is increasing that the experience has positive benefits for family members. In this article, the origin of family presence is described and research evidence about the experience is presented. Three case studies are presented to illustrate typical events, including the potential role of the hospital chaplain. Recommendations for implementation are included. PMID- 15749399 TI - Clinical trials update: sudden cardiac death prevention by implantable device therapy. AB - Sudden cardiac death represents an enormous public health problem in all developed countries of the world. In the United States, sudden cardiac death occurs in more than 400,000 people each year and is the leading cause of death. In sudden cardiac death, the heart abruptly and unexpectedly ceases to function (cardiac arrest), presumably because of an electrical disturbance. Individuals deemed high risk for sudden cardiac death may be treated with implantable defibrillators. This article highlights evidence from randomized, controlled trials of implantable device therapy used in prevention of sudden cardiac death. PMID- 15749400 TI - Documentation of resuscitation events. AB - Resuscitation documentation assists health care professionals in trending patient status, determining what treatments may be most effective, and determining where opportunities for improvement may exist. An overview of what is known about resuscitation documentation is provided in this article, as are implications for future research related to documentation of resuscitation events. Use of the Utstein guidelines in determining essential elements of resuscitation documentation is also presented. PMID- 15749401 TI - Use of handheld devices in critical care. AB - Emergencies happen everyday in the acute care environment and demand nurses to make decisions quickly that can have serious, if not potentially fatal, ramifications. Being prepared to make decisions is partly the results of experience, but having access to ready resources can provide even the newest nurse with the potential to make critical decisions accurately. Handheld devices, such as personal digital assistants, can provide access to resources that can improve patient safety and, ultimately, patient care delivery. PMID- 15749402 TI - Recommendations of the international guidelines 2000 conference on cardiopulmonary resuscitation and emergency cardiac care: an overview. AB - The greatest potential for survival of sudden cardiac arrest can be achieved only by providing early intervention using evidence-based therapies that have been studied over time. Emergency cardiac care and the 2000 advanced cardiac life support guidelines encompass all therapies that have been shown to improve outcomes in patients who experience life-threatening events that involve the cardiovascular, cerebrovascular, and pulmonary systems. Early recognition of warning signs, activation of emergency medical systems within the community, basic cardiopulmonary resuscitation, early defibrillation, airway management, and intravenous medication administration are key factors in improving resuscitation outcomes. PMID- 15749403 TI - Update on pediatric advanced life support guidelines. AB - Accidents are a leading cause of death for children in several nations. Motor vehicle accidents are among the most common causes of cardiac arrest. Other causes include drowning, burns, gunshot wounds, poisoning, smoke inhalation, and airway obstruction caused by asphyxiation from foreign bodies. Approximately 50% to 65% of children who require cardiopulmonary resuscitation are younger than 1 year of age, with most being younger than 6 months of age. To prevent loss and improve quality of life, it is imperative to initiate measures to improve oxygenation and treat cardiac dysfunction. Guidelines for the management of life threatening emergencies in infants and children are internationally similar but not identical. The Australian Resuscitation Council, the American Heart Association, and the European Resuscitation Council current guidelines all have some basic essential techniques for management of pediatric emergencies. PMID- 15749404 TI - Medication safety issues in the emergency department. AB - Medications used in emergency medicine are often "high alert" medications that have to be handled with deliberation. These medications, used every day in the emergency department, are associated with numerous errors. Even newer medications are prone to error. This makes it important for emergency department nurses and health care professionals to develop and implement mechanisms for preventing error. Safety data has shown that these "high alert" medications must be handled appropriately. PMID- 15749405 TI - Use of vasopressors in the treatment of cardiac arrest. AB - Intervention for cardiac arrest may require intervention for electrical abnormalities or hemodynamic instability. These actions can result in ineffective cardiac functioning and systemic hypotension. Vasopressors are capable of improving severe hypotension that can result from reduced cardiovascular contractility or heart rate. These vasopressor actions are critical to successful resuscitation efforts for patients. PMID- 15749406 TI - Pediatric cardiac arrhythmias resulting in hemodynamic compromise. AB - Cardiac arrhythmias in the pediatric population generally do not result in hemodynamic compromise. There are specific scenarios, however, in which an arrhythmia poses a higher risk of deterioration in a child's overall clinical condition. To minimize this risk, clinicians must be able to recognize promptly such arrhythmias, provide rapid clinical assessment, and establish appropriate interventions to avoid cardiac arrest. This article presents a brief overview of hemodynamics in children followed by a discussion of each of the arrhythmias that have a higher-than-usual potential for resulting in hemodynamic compromise. The electrocardiographic characteristics, specific clinical interventions, and nursing implications for each of these arrhythmias also are delineated. PMID- 15749407 TI - The role of thermoregulation in cardiac resuscitation. AB - Regulating a patient's body temperature has long been within the scope of practice of the critical care nurse. Different measures and modalities have been used to achieve normothermia in the past. Recent research has demonstrated how crucial body temperature can be, not only because of its potential for neuroprotection but also because of its effects on all body systems. The general consensus of current literature is that maintaining mild hypothermia at 32 degrees to 34 degrees C (89.6 degrees-93.2 degrees F) for 12 to 24 hours after cardiac arrest may provide optimal neuroprotection with minimal complications for patients. PMID- 15749408 TI - The neurosurgical training curriculum in Australia and New Zealand is changing. Why? AB - The Neurosurgical Advanced Training curriculum of the Royal Australasian College of Surgeons (RACS) is currently undergoing change. Given the high standard of neurosurgery in Australia and New Zealand, it may be questioned why such change is necessary. However, the curriculum has not kept pace with developments in professional practice, educational practice or educational theory, particularly in the assessment of medical competence and performance. The curriculum must also adapt to the changing training environment, particularly the effects of reduced working hours, reducing caseloads due to shorter inpatient hospital stays and restricted access to public hospital beds and operating theatres, and the effects of sub-specialisation. A formal review of the curriculum is timely. PMID- 15749409 TI - Outcome scoring in neurosurgery: the cognitive outcome score, an alternative to the Glasgow outcome scale. AB - The qualitative method, the Glasgow outcome scale, remains the convention for patient outcome assessment in the neurosurgical literature. This is despite the fact that sampling methods subject to bias confer low confidence in the conclusions. An alternative, quantitative method, the cognitive outcome score (COS), decreases bias, is cost effective and delivered in less than 10 min. Utilisation of the COS in statistical correlations against common clinical parameters is discussed in theory and practice. PMID- 15749410 TI - Pediatric pituitary adenoma: a series of 42 patients. AB - Pituitary adenomas are uncommon in childhood. This report describes the presentation, endocrinological profile, management and outcome of 42 children with pituitary adenomas. The majority of the tumors were functioning adenomas (40/42, 95.2%). Only two patients (4.8%) had non-functioning tumors. Amongst the functioning tumors, there were 20 patients (47.6%) with prolactinomas, 11 patients (26.2%) with Cushing's disease and nine patients (21.4%) with growth hormone (GH)-secreting adenomas. The most common presenting features were endocrinopathy and visual disturbances. The majority of tumors were macroadenomas, with only eight patients (all with Cushing's disease) having a microadenoma. Transsphenoidal tumor decompression was performed in most cases (71.4%). Complications following transcranial surgery were more frequent when compared to transsphenoidal surgery. There was one death following surgery. Response to radiotherapy was satisfactory. Overall, 67.6% of patients achieved endocrinological remission. Of these, 89% of the children with GH-secreting tumors and 100% of the children with Cushing's disease achieved remission. We conclude that the transsphenoidal approach is effective and safe in surgery for pituitary adenomas in children and is the procedure of choice if there is no contraindication. The combination of surgery and radiotherapy, as well as medical therapy with bromocriptine, achieves good results in the management of these uncommon tumors. PMID- 15749411 TI - Tiagabine-induced generalised non convulsive status epilepticus in patients with lesional focal epilepsy. AB - PURPOSE: To report 3 cases with focal lesional epilepsy that had non-convulsive status epilepticus (NCSE) induced by treatment with tiagabine (TGB) and review the previously published cases. Drugs that enhance GABAnergic transmission are recognised to promote absence seizures in patients with generalised epilepsy syndromes and may on occasions even induce NCSE. However, that TGB can also induce NCSE in focal lesional epilepsy is not widely recognised in clinical practice. METHOD: The clinical history, EEG and MRI findings were reviewed in 3 patients with lesional focal epilepsy who presented to our epilepsy programs over a 12 month period with TGB-induced NCSE. All previously reported cases in the English medical literature were reviewed. RESULTS: The three patients had longstanding complex partial and secondarily generalised seizures refractory to multiple different anti-epileptic drugs. In two cases, MRI demonstrated a focal malformation of cortical development in the left parieto-occipital region and in the third left mesial temporal sclerosis. Following commencement of TGB in one patient and dose escalation in two, prolonged episodes of confusion and poor responsiveness were noted. Prolonged EEG monitoring demonstrated continuous high amplitude, generalised, 2-4 Hz delta activity with intermingled spikes during the episodes of unresponsiveness, consistent with NCSE. The clinical and EEG activity normalised following the administration of IV clonazepam followed by dose reduction or withdrawal of the TGB. Eleven previously reported cases of patients with partial epilepsy and a focal underlying lesion on MRI were identified, all of whom had similar features to that seen in our cases. CONCLUSIONS: These cases illustrate that TGB may induce generalised NCSE in patients with focal lesional epilepsy, in addition to those with generalised syndromes. We hypothesise that patients may have developed an acquired alteration in the sensitivity of their thalamocortical circuitry that renders them more sensitive to the effects of drugs that enhance GABAnergic activity. PMID- 15749412 TI - Electrophysiological, clinical and epidemiological study of Guillain-Barre Syndrome in Hong Kong Chinese. AB - The authors reviewed the clinical and electrophysiological features in 20 consecutive adult patients with Guillain-Barre Syndrome (GBS) admitted to a regional hospital in Hong Kong from 1993 to 1998. The majority of cases in this locality consists of the demyelinating form of GBS; epidemic, acute motor axonal neuropathy is not the predominant form. The incidence of GBS in this region of China was 0.44 per 100,000. PMID- 15749413 TI - Effect of GPi DBS on functional imaging of the brain in dystonia. AB - Five patients with idiopathic dystonic conditions, treated successfully with deep brain stimulation (DBS) of the globus pallidus internus (GPi), were studied using single-photon emission tomography (SPET) in order to evaluate brain perfusion in the presence and absence of DBS. Comparison was made between the "on" and "off" DBS scans on an individual basis and also as part of a group analysis. Whilst the individual data suggested great regional variation in cerebral perfusion between individuals, the results of the group analysis revealed several topographically similar areas of the brain where relative hyperperfusion in the absence of DBS was common to all patients. Based on these results we postulate on possible mechanisms for this phenomenon. PMID- 15749414 TI - CSF spectrophotometry in the diagnosis and exclusion of spontaneous subarachnoid haemorrhage. AB - AIMS: To assess the sensitivity and specificity of CSF spectrophotometry for the detection of xanthochromia in patients with clinical symptoms suggestive of subarachnoid haemorrhage (SAH) but normal cranial computed tomography (CT). METHODS: All consecutive patients undergoing both cranial CT and lumbar puncture for investigation of possible SAH at the Princess Alexandra Hospital, Brisbane, between January 2000 and April 2003 were included in the study. All case histories, radiology and laboratory results were retrospectively assessed. The sensitivity, specificity and positive predictive value of the spectrophotometry test were calculated. RESULTS: 253 patients were included in the study. Spectrophotometry was shown to have a sensitivity of 100% but a specificity of only 75.2%. The positive predictive value of spectrophotometry as an indicator of SAH was 3.3%. CONCLUSIONS: CSF spectrophotometry has an unacceptably low specificity and positive predictive value, which greatly limit its use as a clinical tool. PMID- 15749415 TI - Non-shaved ventriculoperitoneal shunt in Thailand. AB - The objective of this study was to determine whether non-shaved patients undergoing ventriculoperitoneal shunt operations would experience a significantly increased shunt infection rate as compared to shaved patients. Clinical trials of non-shaved scalp preparation were performed in ventriculoperitoneal shunt procedures at Songklanagarind Hospital from January 1994 to December 1999. Exclusion criteria were poor scalp condition, previous shunt procedures and immunocompromised hosts. Statistical analysis using univariate, multivariate and logistic regression was used. One hundred and nineteen patients were included in the study. Thirty-six cases were in the non-shaved group. Thirty-eight of 119 cases were less than one year of age and congenital hydrocephalus was the most common problem. The only notable risk factor for shunt infection was an operation lasting more than 59 min. The shunt infection rate in the non-shaved and shaved groups was 6.25% and 14.94%, respectively (p>0.05). In conclusion, leaving the hair intact for ventriculoperitoneal shunt surgery is safe and not associated with increased risk of shunt infection. The non-shaved method may encourage quicker restoration of the patients' self-image and facilitate their early return to normal life. PMID- 15749416 TI - Postoperative vasospasm of unruptured paraclinoid carotid aneurysms: analysis of 30 cases. AB - This study was conducted to determine the incidence, severity, and causes of delayed vasospasm after clipping of unruptured paraclinoid aneurysms of the internal carotid artery (ICA). A retrospective analysis was made of 30 patients, who underwent clipping of unruptured paraclinoid aneurysms in our institution between 1991 and 1998. We compared angiograms before and after operation and classified them into two groups: vasospasm group and non-vasospasm group. Eleven variables were assessed as to their relationship to delayed vasospasm. There were 9 patients (30%) in the vasospasm group, of which 3 patients (10%) were clinically symptomatic. For all symptomatic patients, aggressive treatment, including triple-H therapy, was conducted with good outcome. The number of clips used (p<0.04) and temporary occlusion of the ICA (p<0.005) were statistically significant factors associated with the incidence of vasospasm. It is suggested that mechanical stimulation to the vascular wall of the ICA is responsible for causing spasm in addition to intraoperative bleeding around the dural ring. PMID- 15749417 TI - Two-point discrimination following traumatic brain injury. AB - This study investigated two point discrimination (TPD) perception in survivors of traumatic brain injury (TBI). Inpatient and outpatient survivors of severe TBI and age-matched healthy controls aged between 16 and 65 years were included in the study with a mean TPD of 3.61, 3.41 and 2.61 mm respectively. Significant group effects were seen in TPD between subjects with TBI and controls. TPD deficits did not appear to be influenced by GCS or PTA duration, nor did they show evidence of improvement over time. Similarly, CT scan data did not explain the observed TPD differences in TBI survivors. Admission functional independence measure (FIM), a global measure of functional independence, had a strong negative correlation with TPD. The lack of change in TPD over time mirrors other basic markers of neurological recovery but is at odds with TBI outcome literature reporting continuing improvements in function for at least 5 years post injury. PMID- 15749418 TI - A proposed mechanism for diurnal/nocturnal bruxism: hypersensitivity of presynaptic dopamine receptors in the frontal lobe. AB - There are many reports in the literature concerning nocturnal bruxism, however, diurnal (non-sleep)/nocturnal bruxism is rarely mentioned. We report three patients with diurnal/nocturnal bruxism. They differed from the usual features of nocturnal bruxism in hypoperfusion of the left frontal lobe, a poor response to l dopa or bromocriptine therapy and a favourable response to metoclopramide. Hypersensitive presynaptic dopamine receptors may be the underlying pathology responsible for this type of bruxism. Regional differences in dopamine receptor pharmacology may explain the perplexing relationship of bruxism to both hyper- and hypo-dopaminergic states. PMID- 15749419 TI - Images in neuroscience. PMID- 15749420 TI - The effect of cyclin D expression on cell proliferation in human gliomas. AB - The expression of three cyclin D subtypes was defined immunohistochemically with cyclin D1, D2 and D3 monoclonal antibodies in 52 human glioma biopsies and eight control samples of normal brain tissue. PCNA labeling indices (LI) were used to evaluate proliferation in the glioma biopsies. LI of cyclin D1, D2 and D3 were compared with histological grade and the proliferating cell nuclear antigen (PCNA) LI. Cyclin D1 expression only was observed in normal brain tissue, but marked overexpression of cyclin D1 and cyclin D3 was observed in glioma. Cyclin D1 LI increased with malignancy, in parallel with an increase in PCNA LI. Lower expression of cyclin D2 was found in a small fraction of the gliomas, but its LI did not vary significantly with grade. Cyclin D3 was mainly expressed by malignant gliomas and was rarely observed in low-grade glioma. Cyclin D2 and D3 expression correlated with PCNA LI, but not as strongly as for cyclin D1. Expression of cyclin D1 is closely related to both the oncogenesis and progression of glioma, while cyclin D3 is associated with transformation to a malignant phenotype. Cyclin D2 is weakly expressed and shows no marked relationship with any aspect of tumorigenesis. The exact contribution of cyclin D subtypes to cell cycle progression in neoplastic and reactive cells remains to be defined. PMID- 15749421 TI - Bilateral decompression of multilevel lumbar spinal stenosis through a unilateral approach. AB - Lumbar canal stenosis due to hypertrophy and calcification of the facet joints and/or ligamentum flavum is a common condition in the elderly. Although a large number of individuals are symptom-free, the degenerative process, usually encroaching on both central and lateral pathways, may lead to symptoms of itself or decompensate a preexisting narrow canal. Even at an advanced age, decompression surgery is effective for symptomatic stenosis. Less invasive procedures preserving maximal bony and ligamentous structures have recently been recommended to reduce associated morbidity. This paper introduces a unilateral surgical approach for bilateral decompression by ligamentectomy, partial facetectomy and foraminal unroofing. Using a specially designed, one-side retractor, after the ipsilateral nerve root decompression the contralateral dural sac and nerve roots were approached through an 8 x 15 mm window in the interspinous ligament. The contralateral ligamentum flavum, facet joints and foraminal roof were resected, preserving the supraspinous ligament complex and much of the contralateral musculature. This technique, preserving anatomy and biomechanical function of the lumbar spine, is useful for surgery on multilevel lumbar canal stenoses. PMID- 15749422 TI - Trigeminal neuralgia in a patient with mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). AB - Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disease associated with multiple deletions of mitochondrial DNA in skeletal muscle. MNGIE is a multisystem syndrome affecting muscle, peripheral, and central nervous systems and the gastrointestinal tract. A 25-year-old man is presented with 3 years history of right sided trigeminal neuralgia. He has been diagnosed as MNGIE based on clinical, neurophysiological and pathological findings. He had also received medical therapy and two radiofrequency thermocoagulations for the treatment of trigeminal neuralgia. Gamma Knife radiosurgery was performed and resulted in partial relief. To our knowledge, this is the first case in the literature of MNGIE with trigeminal neuralgia. An analogy is suggested between multiple sclerosis and MNGIE as a cause for trigeminal neuralgia in this patient. PMID- 15749423 TI - "Fou rire prodromique" associated with simultaneous bilateral capsular genu infarction. AB - Pathological laughter is exaggerated, uncontrollable, and inappropriate laughter usually unrelated to a true emotion or a congruent mood. "Fou rire prodromique" is a rare form of prodromal pathological laughter of uncertain pathophysiology that heralds an ischaemic neurologic deficit. We report a case of prodromal pathological laughter marking the onset of bilateral capsular genu infarction. T2 weighted cranial magnetic resonance imaging (MRI) showed bilateral capsular genu infarction. There was also a diffusion defect in the same areas on diffusion weighted imaging (DWI). Although it is known that bilateral subcortical lesions can cause pathological laughter, this is the first demonstration of simultaneous associated bilateral capsular genu infarction on cranial DWI MRI. PMID- 15749424 TI - Functional imaging of a large demyelinating lesion. AB - PURPOSE: To determine the metabolic characterization of a large solitary demyelinating lesion. METHODS: Magnetic Resonance Spectroscopy (MRS) and Positron Emission Tomography (PET) studies with 2-deoxy-2-[F-18]fluoro-d-glucose (FDG), carbon-11-methionine (methionine) and carbon-11-choline (choline) were done on the demyelinating lesion. RESULTS: The demyelinating lesion exhibited a low glucose uptake, prominent methionine uptake and a minimal choline uptake on the PET studies. MRS data revealed an increased choline to creatine (cho/cr) ratio and a decreased N-acetyl-aspartate to creatine (NAA/cr) ratio, which demonstrated a return to near normal ratios on follow-up study. CONCLUSION: The report summarizes the metabolic characteristics of a demyelinating plaque. PMID- 15749425 TI - Aqueduct stenosis-?Benign. AB - 'Benign' aqueduct stenosis is a common cause of hydrocephalus in the paediatric population and is frequently treated by endoscopic third ventriculostomy. Occasionally, aqueduct stenosis can be a prelude to the development of other pathology, as is seen in these two cases of pineal tumours developing in patients whose hydrocephalus was successfully treated with endoscopic third ventriculostomy. The case histories are presented, along with the recommendation for increased radiological screening of patients with this usually 'benign' presentation. PMID- 15749426 TI - Posterior temporal haematoma associated with anterior communicating artery aneurysm rupture. AB - We report the case of a 68-year-old man who presented with symptoms suggesting an intracranial haemorrhage. A computed tomography scan showed subarachnoid haemorrhage associated with a remote posterior temporal intracerebral haematoma. Angiography revealed the presence of an anterior communicating artery aneurysm, which was subsequently clipped. Possible causes for the association of a distant intracerebral haematoma with rupture of an anterior communicating artery aneurysm are discussed. This uncommon association should be recognised to avoid incorrect identification of the origin of haemorrhage. PMID- 15749427 TI - Metastatic lung cancer in the cerebellopontine angles mimicking bilateral acoustic neuroma. AB - Bilateral cerebellopontine angle (CPA) tumors identified on MRI are considered bilateral acoustic neuromas, the definitive diagnostic criterion of neurofibromatosis 2 (NF-2). We report the case of a 67-year-old man with progressive bilateral hearing loss, vertigo, and imbalance. MRI revealed bilateral enhancing CPA lesions, which were suggestive of acoustic neuromas and a diagnosis of NF-2. However, autopsy showed metastatic adenocarcinoma of the lung. Therefore, metastatic carcinoma to the CPA can mimic bilateral acoustic neuromas; imaging studies alone may be insufficient to diagnose NF-2. PMID- 15749428 TI - Cerebral venous thrombosis in a gentleman presenting with fever, convulsion and frontotemporal haemorrhages. AB - Cerebral venous thrombosis (CVT) is an uncommon but serious type of stroke. Thrombosis may involve the cortical or deep veins or the venous sinuses. The presenting clinical features are non-specific. We report a 48-year-old man with CVT who presented with fever, bitemporal throbbing headache, and generalised convulsion. Computed tomography (CT) of the brain revealed acute haemorrhages over right anterior frontal and posterior temporal regions with surrounding oedema and right anterior temporal subcortical oedema. The initial diagnosis was herpes simplex encephalitis. Absence of venous flow over the right transverse and sigmoid sinuses during the venous phase of digital subtraction angiography (DSA) revealed CVT. He was anti-coagulated for 6 months. An underlying cause of CVT was not detected. A high index of suspicion is required when risk factors of CVT are present. CT brain may be normal or showing non-specific findings. Magnetic resonance imaging plus venography, CT venography, or DSA is diagnostic. PMID- 15749430 TI - Pituitary abscess in a patient with painless thyroiditis. AB - The preoperative diagnosis of pituitary abscess is difficult. The case of a 56 year-old man with a pituitary abscess and painless thyroiditis is presented. There has been no previous such report. The differential diagnosis includes pituitary abscess, lymphocytic adenohypophysitis or infundibuloneurohypophysitis, share clinical symptoms of panhypopituitarism and diabetes insipidus, and is of critical importance as the treatment of these conditions differ. The association with painless thyroiditis suggests a diagnosis of lymphocytic adenohypophysitis or infundibuloneurohypophysitis. However, the coincidence of pituitary abscess and painless thyroiditis was observed in our patient, and thus though rare, should be considered. Surgical exploration and histopathological examination are essential for the differential diagnosis of these diseases. PMID- 15749429 TI - Vertically acquired neonatal citrobacter brain abscess - case report and review of the literature. AB - Vertically acquired citrobacter meningitis in the neonate is very rare and carries a very high mortality and morbidity. Overall, approximately 30% of neonates with Citrobacter meningitis die and 50% sustain some damage to the CNS. The authors describe a case of a newborn with Citrobacter koseri meningitis with multiple brain abscesses, with a successful outcome following multiple burr-hole aspirations and prolonged antibiotic therapy. An aggressive surgical approach combined with intravenous antibiotics (including imipenems, to which the organism is very sensitive) for a minimum of 4 weeks appears to improve the outcome of infection with this virulent organism. PMID- 15749431 TI - Post-traumatic cervical kyphosis with surgical correction complicated by temporary anterior spinal artery syndrome. AB - Post-traumatic undiagnosed disco-ligamentous and osseous lesions of the cervical spine may eventually result in irreducible extreme kyphosis. Correction of such consolidated deformities requires major surgery with a combined posterior and anterior approach, aiming to correct bony impingement on neural and vascular structures, reduce deformity and to attain circumferential instrumentation and fusion in physiological alignment. This can be achieved using either a single staged or a two-staged procedure. Regardless, this type of major surgery entails considerable neurological risks. Therefore, thorough planning of the intervention and considerable surgical experience is needed. We present an elderly woman with gross restriction of forward gaze and intractable nuchal and radicular pain due to cervical spine deformity. Her cervical kyphosis was corrected using preoperative skeletal axial traction for four days and subsequent operative reduction with circumferential instrumentation and fusion. The post-operative course was complicated by a temporary anterior spinal artery syndrome despite normal intraoperative somatosensory evoked potentials (SSEP) and by a wound infection requiring removal of the implant. Nevertheless, segmental fusion in physiological alignment was successfully achieved and the patient fully recovered from the neurological deficit and infection. Quality of life was significantly improved. PMID- 15749432 TI - Conservative management of spontaneous posterior fossa subdural hematoma in a neonate. AB - Symptomatic intracranial hemorrhage in term neonates is uncommon, and is usually secondary to trauma, coagulation disorders and/or hypoxia. Posterior fossa hemorrhage in the neonate is a rare neurosurgical emergency and is usually associated with the same etiological factors. Diagnosis is with computed tomography and magnetic resonance imaging. We present a spontaneous posterior fossa subdural hematoma in a term neonate and discuss conservative management. PMID- 15749433 TI - Ruptured "giant" supratentorial dermoid cyst. AB - Intracranial dermoid and epidermoid cysts are rare lesions formed from the inclusion of ectodermal elements during neural tube closure. Although not entirely consistent, imaging characteristics on CT and MRI can aid differentiation of dermoids and epidermoids, as can age at presentation, location and tendency to rupture. The distinction between dermoid and epidermoid lesions is important prognostically and may impact on surgical management as a subtotally resected dermoid is less likely to recur than its epidermoid counterpart. The distinction of large dermoid lesions as "giant" adds little to information regarding the natural history or prognosis of these lesions and should be abandoned. PMID- 15749434 TI - Treatment of empty sella syndrome with ventriculoperitoneal shunt. AB - A symptomatic empty sella developed in a female patient undergoing bromocriptine therapy for microprolactinoma. Placement of a ventriculoperitoneal shunt dramatically improved the symptoms of headache and blurred vision. The post operative imaging showed resolution of the empty sella. She was able to resume bromocriptine therapy without recurrence of her previous symptoms and give birth to a baby 20 months later. An MRI 44 months after surgery and on bromocriptine therapy showed no recurrence of the empty sella. We conclude that ventriculoperitoneal shunt may be a simple, and durable treatment for drug induced empty sella and allows resumption of bromocriptine therapy for preexisting microprolactinoma. PMID- 15749435 TI - Progressive severe anemia due to copper deficiency five years after subarachnoid hemorrhage. AB - Patients with neurological illnesses often require prolonged enteral or parenteral nutrition. The complications of this type of feeding may be unfamiliar to neurosurgeons and neurologists, however, these complications may be an important source of ongoing morbidity. We present a case of severe anemia due to copper deficiency in a patient requiring enteral nutrition after SAH. The anemia resolved with copper supplements. A high index of suspicion is needed to recognise and treat complications of enteral nutrition. PMID- 15749436 TI - Orbitofrontal cholesterol granuloma. AB - Cholesterol granuloma of the orbital bones is a rare but readily recognisable condition. It is an osteolytic lesion with a granulomatous reaction surrounding cholesterol crystals, old haemorrhage and a fibrous capsule. There is a male preponderance and it usually occurs in young or middle-aged men. It is treatable with drainage and curettage via an orbitotomy, and craniotomy or wide bone removal is almost never required. Six cases of this condition were reviewed to highlight the typical clinical presentation, computed tomography and magnetic resonance results, and surgical management. PMID- 15749437 TI - Letter to the neurology editor. PMID- 15749438 TI - Letter to the editor. PMID- 15749439 TI - How is the tibial tray aligned to the femoral prosthesis in a total knee arthroplasty? A survey of opinion from BASK? PMID- 15749440 TI - Patellofemoral forces after total knee arthroplasty: effect of extensor moment arm. AB - Total knee arthroplasty implant designs with larger extensor moment arms theoretically should generate lower extensor forces for the same externally applied loads. This study measured knee kinematics, quadriceps forces, and patellofemoral forces under conditions of dynamic knee extension under load in two knee designs with differing quadriceps moment arms. Six human cadaver knees were tested both before implantation and after sequential implantation with two posterior cruciate retaining designs. The extensor moment arm of the LMA (long extensor moment arm design, Scorpio, Howmedica Osteonics, Rutherford, NJ) was approximately 1 cm longer than that of the Control design (7000, Howmedica Osteonics). Quadriceps tension was measured during dynamic closed kinetic chain knee extension. Patellar compressive and shear forces were also recorded using a patellar component instrumented with a custom triaxial load transducer. Knee kinematics were monitored using a three-dimensional electromagnetic tracking device. Both designs produced similar patterns of femoral rollback and tibial rotation. Quadriceps tension was lower in the LMA design compared with the Control design. Patellofemoral compressive forces were also significantly reduced in the LMA design when compared with Control (8-18% lower at angles greater than 50 degrees flexion). The design with the longer extensor moment arm required less quadriceps force to extend the knee under load and reduced patellofemoral compressive forces. Reduced quadriceps forces may facilitate postoperative rehabilitation and activities such as stair climbing. Reduction in patellofemoral forces could reduce patellar complications such as anterior knee pain, component wear, and loosening. PMID- 15749441 TI - The use of the knee joint-line balancer to control patella position in revision total knee arthroplasty. AB - In revision total knee arthroplasty (TKA), control of the patellar height and the joint line is difficult. Therefore, we developed an adjustable flexion-extension spacer, the Knee Joint-line Balancer (KJB(R)). This device simulates femur component sizes, polyethylene sizes, the joint-line level, and distal femur wedges. The goal of this study is to evaluate the use of an adjustable knee spacer to control patellar height and joint-line during revision total knee surgery. The subjects of the study were the first 10 consecutive patients who had undergone revision of a primary TKA where the KJB was used. A reference group composed of the last 10 patients treated without the use of the KJB was also evaluated. The joint-line position and the patellar height were determined before and after revision TKA. The method described by Figgie et al. was used. The patellar height in the reference group averaged 7.7 mm. Seven of 10 patients had a patella baja, and two of these patients had patellar impingement. One patient needed a proximalisation of the tuberositas. The patellar height in the KJB group averaged 14.6 mm after revision, with only one patient having a patella baja. This new device, adjustable kneespacer for revision TKA ("the KJB"), seems to provide better control of the patellar position in total knee revision. PMID- 15749442 TI - A new technique for reconstructing the medial patellofemoral ligament. AB - Many surgical techniques exist for treating patella instability. Over recent years, attention has focussed on reconstruction of the medial patellofemoral ligament (MPFL). We report a procedure involving a lateral release, distal realignment of the tibial tubercle and anatomic reconstruction of the MPFL using a graft composed of a redundant strip of medial retinacular tissue. Nineteen patients (twenty-one knees) with chronic patellofemoral instability who had failed conservative treatment underwent this procedure and were assessed clinically and radiologically at an average of 23 months post-operation. No patients were lost to follow-up. There was no recurrence of subluxation or dislocation. All achieved levels of activity comparable to or improved compared to pre-operative uninjured levels. This combined procedure incorporating a previously un-described method of reconstructing the MPFL gives superior results compared to previous studies. No graft donor site morbidity occurred and patient outcome scores, especially objective stability of the patella to lateral shift, were excellent. PMID- 15749443 TI - A safe zone for the passage of screws through the posterior tibial cortex in tibial tubercle transfer. AB - In tibial tubercle transfer, surgery drills and screws can put the popliteal vessels at risk if the posterior cortex is breached. This complication can be devastating. We have looked at arteriograms of 50 knees and identified a safe zone through which an instrument can be passed with more confidence. In our study we found no vessels directly posterior to the supero-medial aspect of the proximal metaphysis in any knee. Whilst care must still be taken, this area will allow surgeons greater confidence to obtain a stronger bicortical hold with any fixation device. PMID- 15749444 TI - Arthrofibrosis following total knee replacement; does therapeutic warfarin make a difference? AB - Arthrofibrosis following total knee replacement (TKR) is a relatively common complication which results in a reduction in knee range of movement and patient dissatisfaction. A retrospective study examined the relationship between anticoagulation with therapeutic warfarin and rates of arthrofibrosis following TKR. Arthrofibrosis was defined as less than 80 degrees of knee flexion 6-8 weeks post-TKR. Patients were warfarinised if they had a history of thrombophilic tendencies or medical conditions necessitating anti-coagulation, rather than as routine thromboprophylaxis. All other patients received thromboprophylaxis using low molecular weight heparin. A total of 728 patients underwent 874 primary TKR between 1993 and 2002 in one centre, performed by four surgeons. Mean age was 68 years (range 48-89 years) and there were 483 female and 391 male knees. Eighty cases were warfarinised post-operatively (53 female, 27 male). Overall, 83 of 874 TKRs (9%) had arthrofibrosis (57 female, 26 male) requiring manipulation under anaesthetic (MUA). In the warfarinised group, 21 knees (26%) had an MUA (15 female, 6 male). This compared to 62 cases (8%) requiring MUA in the non warfarinised group (42 female, 20 male). There was a statistically significant difference on Fisher's exact testing (P<0.0001) between groups. Following MUA, knee flexion improved in 95% cases to a minimum 95 degrees but 8 cases had a fixed flexion deformity of 5-10 degrees . In conclusion, therapeutic warfarinisation post-TKR leads to a statistically greater chance of the patient developing arthrofibrosis compared to prophylactic low molecular weight heparin and that patients should be counseled appropriately. PMID- 15749445 TI - The effect of aprotinin on blood loss in bilateral total knee arthroplasty. AB - Twenty-five patients undergoing bilateral total knee arthroplasty were given aprotinin (1-2 million units) in doses adjusted for weight, infused at the end of the first arthroplasty. Blood management was compared with our previous series of 68 patients who were not treated with aprotinin. Blood drained and re-transfused in the first 6 h was significantly less in the aprotinin group (323 ml, S.D. 320) than the untreated group (1033 ml, S.D. 539, p<0.0005). The total blood drained was also less in the aprotinin group (713 ml compared to 1613 ml, p<0.0005) but this was accounted for soley by the difference in early blood loss. The use of other blood products was less in the aprotinin group (360 ml, S.D. 357 compared to 827 ml, S.D. 434, p<0.0005). The mean postoperative haemoglobin on the day following surgery was similar at around 10 g. There were no complications or adverse reactions to the use of aprotinin. We conclude that aprotinin significantly reduces early blood loss after bilateral knee arthroplasty. In our hands, we have found it to be safe. Further work is required to determine whether postoperative use may further reduce the need for blood transfusion. PMID- 15749446 TI - Role of insulin like growth factor-I in repair response in immature cartilage. AB - OBJECTIVE: The purpose of this study was to investigate the effects of exogenous local Insulin like growth factor-I (IGF-I) on the repair of full-thickness articular cartilage defects in immature rabbits. DESIGN: Thirty-six skeletally immature New Zealand rabbits between 6 and 8 weeks old were used. A single defect, 3.5-mm-wide by 4-mm-deep full-thickness articular cartilage defect in the medial femoral condyle, was created. The defect was either filled with a collagen sponge or with a collagen sponge impregnated with 5 mug of recombinant IGF-I. The animals were sacrificed at 4, 8 or 12 weeks, and the repair tissue was examined macroscopically and histologically. Repair tissue was also examined immunohistochemically for the presence of type-I collagen, type-II collagen and PCNA at all weeks. RESULTS: Newly formed tissue in all of the defects in the IGF I group had the gross, histological and histochemical appearance of a smooth, intact hyaline articular cartilage. The average total scores on the histological grading scale were significantly better (p<0.05) for the defects treated with recombinant IGF-I at all time points. Immunostaining with an antibody against type-II collagen showed the diffuse presence of the repair cartilage in the IGF-I treated defects. The control groups demonstrated minimum staining with type-II collagen antibody. CONCLUSIONS: These findings suggest that repair of full thickness immature cartilage defects can be enhanced by recombinant IGF-I. PMID- 15749447 TI - Gait and clinical measurements in patients with knee osteoarthritis after surgery: a prospective 5-year follow-up study. AB - The aim of this prospective follow-up study was to determine if gait measurements and/or clinical measurements could detect differences in treatment outcome between two surgical interventions in patients with knee osteoarthritis (OA). The patients were followed for 5 years after surgery. Forty patients, 55-70 years of age, with unilateral knee OA were included. The patients were treated either with a high tibial osteotomy (HTO) (n=18) or a unicompartmental knee arthroplasty (UKA) (n=22). Clinical outcome measures were the British Orthopaedic Association (BOA) score, pain during walking, passive range of knee motion (PROM) and patients' subjective opinion. The gait variables were free walking speed, step frequency, step length and single and double-stance phase for each leg. The patients were examined before surgery and 3 months, 1 year and 5 years after surgery. The time-distance variables of gait could detect differences in treatment outcome, 3 months after surgery, while the clinical outcome measures, as given here, could not detect any differences between the two groups of patients. Measurements of free walking speed could be recommended for clinical evaluation, after surgical interventions, in patients with knee OA. PMID- 15749448 TI - Electromyographic analysis of the knee during jump landing in male and female athletes. AB - Many noncontact anterior cruciate ligament (ACL) injuries in female athletes occur at foot strike during jump landing when the knee is extended. This study was undertaken to determine the activation level of the quadriceps and hamstring muscles electromyographically. Fifteen healthy volunteers (eight women and seven men), all of whom were collegiate basketball players, participated in the study. The maximum voluntary contraction (MVC) of the vastus medialis (VM) at a knee flexion angle of 15-45 degrees was significantly higher in women than in men. There was no significant difference in overall mean hamstring activity in men and women over the same knee flexion range. However, when the knee flexion angle was 15 degrees , 20 degrees , and 25 degrees , hamstring activity was significantly lower in female athletes. These results suggest that female athletes have a higher risk of ACL injury during jump landing due to increased anterior tibial translation force with quadriceps muscle activity. Female athletes require greater hamstring activation, and it is suggested that exercising this muscle will increase its activity when the knee is extended, thus preventing ACL injury during actual sport motions. PMID- 15749449 TI - Mechanical analysis of fixation methods for anterior cruciate ligament reconstruction with hamstring tendon graft. An experimental study in sheep knees. AB - This study evaluates the structural properties of the femur-anterior cruciate ligament (ACL) graft-tibia complex, comparing different graft fixation techniques in sheep knees. Four fixation devices were tested both for femoral fixation (Transfix, absorbable screw, RCI screw and Linx-HT) and tibial fixation (bone plug with metal screw, absorbable screw with staple, RCI screw and cancellous screw with spiked washer). The graft used for ACL reconstruction was fresh ovine doubled Achilles tendon (DAT). Femurs and tibias were tested separately. Two mechanical test series were performed on the specimens: a load-to-failure test and a cyclic loading test. On the femoral side, transcondylar screw showed the greatest fixation strength and stiffness and the lowest elongation at cyclic loading. Tibial fixation complexes seemed to have poorer structural properties in comparison to femoral fixation. Among the tibial fixation devices, absorbable screw with staple fixation showed the greatest strength and stiffness. Spiked washer fixation showed the greatest elongation under cyclic loading. PMID- 15749450 TI - Reconstruction of ruptured patellar tendon after total knee arthroplasty: a case report and a description of an alternative fixation method. AB - Patellar tendon rupture after total knee arthroplasty is a rare, but often catastrophic complication. Many different reconstruction techniques of patellar tendon rupture have been described with variable and often discouraging results. A case report with patellar tendon rupture after a total knee arthroplasty is presented. Also, an alternative surgical technique of reconstruction of the patellar tendon using a semitendinosus-gracilis (STG) graft with an interference screw and a staple fixation enabling an immediate mobilization is described. PMID- 15749451 TI - Above-knee Ilizarov external fixation for early periprosthetic supracondylar femoral fracture--a case report. AB - Supracondylar femoral fractures above a total knee replacement are rare injuries that may be challenging to treat. We present a 60-year-old woman who sustained a supracondylar femoral fracture 10 days following a total knee replacement. This patient had multiple risk factors. The fracture was not deemed amenable to conventional treatment. This patient underwent fixation of her femoral fracture above a total knee replacement using a two-ring above-knee Ilizarov external fixator. This allowed full mobilization of the affected limb during fracture healing. The fixator was removed at 10 weeks, at which time the fracture was solidly healed. At the most recent follow-up, 14 months from injury, she was fully weight-bearing without walking aids and had a knee range of motion of 0-120 degrees . PMID- 15749452 TI - Investigation of a hybrid method of soft tissue graft fixation for anterior cruciate ligament reconstruction. AB - To increase knee stability following anterior cruciate ligament (ACL) reconstruction, development of increasingly stronger and stiffer fixation is required. This study assessed the initial pullout force, stiffness of fixation, and failure modes for a novel hybrid fixation method combining periosteal and direct fixation using porcine femoral bone. A soft tissue graft was secured by combining both an interference screw and an EndoButton (Smith and Nephew Endoscopy, Andover, MA). The results were compared with the traditional direct fixation method using a titanium interference screw. Twenty porcine hindlimbs were divided into two groups. Specimens were loaded in line with the bone tunnel on a materials testing machine. Maximum pullout force of the hybrid fixation (588+/-37 N) was significantly greater than with an interference screw alone (516+/-37 N). The stiffness of the hybrid fixation (52.1+/-12.8 N/mm) was similar to that of screw fixation (56.5+/-10.2 N/mm). Graft pullout was predominant for screw fixation, whereas a combination of graft pullout and graft failure was seen for hybrid fixation. These results indicate that initial pullout force of soft tissue grafts can be increased by using the suggested novel hybrid fixation method. PMID- 15749453 TI - Hepatotoxicity and metabolism of trabectedin: a literature review. AB - Trabectedin is a promising anticancer drug currently undergoing phase II evaluation. In preclinical studies, trabectedin was found to cause hepatotoxicity and in patients it reversibly increases plasma levels of liver enzymes. On the basis of preclinical work, it was suggested that metabolism of trabectedin contributed to the pharmacological effects of trabectedin, including hepatotoxicity in rats and increases in liver enzymes in humans. Our aim was to review the current literature on the metabolism of trabectedin and its role in the increases in liver enzymes and hepatotoxicity. We conclude that the trabectedin metabolic profile appears to predict the reversible nature of hepatotoxicity. The rat may not be the best species to investigate trabectedin hepatotoxicity because both trabectedin metabolic profile and reversibility of hepatotoxicity differs from humans. Humans and monkeys display a similar metabolic profile of trabectedin and in both species hepatotoxicity is reversible. Trabectedin is a drug with predictable hepatotoxic effects. Monitoring of plasma levels of liver enzymes ensures safe use of trabectedin in the clinic. Future investigations must be aimed at elucidating the mechanism of trabectedin hepatotoxicity. PMID- 15749454 TI - 150 kDa glycoprotein isolated from Solanum nigrum Linne enhances activities of detoxicant enzymes and lowers plasmic cholesterol in mouse. AB - This study was carried out to investigate the modulation of detoxicant enzyme activity and plasma lipidemic levels by 150 kDa glycoprotein isolated from Solanum nigrum Linne (SNL), which has been used as a hepatoprotective and anticancer agent in folk medicine. Our results in this study showed that SNL glycoprotein has a band with 150 kDa on the 10% sodium dodecyl sulfate polyacrylamide gel, and that it has a strong scavenging activity against lipid peroxyl radicals. We also evaluated the lipidemic levels of SNL glycoprotein, based on lipoproteins and activities of detoxicant enzymes in treatment with Triton WR-1339 or corn oil in vivo. When mice were treated with either Triton WR 1339 or corn oil in the absence of SNL glycoprotein, the number of plasma lipoproteins [triglyceride (TG), total cholesterol (TC) and low density lipoprotein (LDL)] increased. However, when the mice were treated with either Triton WR-1339 or corn oil in the presence of SNL glycoprotein, the plasma lipoprotein levels (TG, TC and LDL) were significantly reduced. Similar results of SNL glycoprotein treatment were also produced in the activities of detoxicant enzymes. Namely, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were remarkably increased after treatment with SNL glycoprotein. In addition, the activity of hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was reduced by SNL glycoprotein in cholestyramine treated mice. For example, we found that it inhibits the activity of cholestyramine-induced hepatic HMG-CoA reductase at 40 microg head body weight g( 1) SNL glycoprotein. Collectively, these results pointed out that SNL glycoprotein can enhance the activities of detoxicant enzymes and bring about the inhibition of HMG-CoA reductase activity in vivo. Therefore, we speculate that SNL glycoprotein can be used as a cholesterol-lowering agent even at low concentrations. PMID- 15749455 TI - Effects of endotoxic shock on neuronal NOS and calcium transients in rat cardiac myocytes. AB - OBJECTIVE: The effects of endotoxic shock on transcriptional and translational pattern of nitric oxide synthase isoforms (NOSs) and cytoplasmic calcium were investigated. METHODS: Male SD rats injected with lipopolysaccharides or saline were sacrificed after 6 and 20 h. Cardiac myocytes were enzimatically isolated from the excised hearts and evaluated for: (1) expression of constitutive (e and n) and inducible (i) NOSs by RT-PCR; (2) NOSs protein levels by Western blot, enzymatic activities by a radioimmunometric assay and nitric oxide metabolites by spectrophotometry; (3) calcium transients by Indo-1 fluorescence. RESULTS: Increase in iNOS mRNA, and decrease in e and nNOS mRNAs were observed in cardiac myocytes isolated 6h after LPS injection with recovery to basal levels at 20 h. Significant down-regulation of e and nNOS protein levels (p < 0.01) and calcium dependent activity (p < 0.05) were detected at 20 h. Serum TNF-alpha increased after 6 and 20 h (p < 0.05), whereas NO metabolites rose only after 20 h (p < 0.0001). The diastolic calcium increased 6 h from LPS injection (p < 0.0001) and remained significantly higher after 20 h. Calcium transients amplitude was not affected by LPS injection. CONCLUSIONS: Endotoxic shock stimulates iNOS and down regulates expression of nNOS in purified cardiac myocytes, but endogenous NO production does not likely affect calcium transients. PMID- 15749456 TI - Serotonin 5-HT2C receptor-mediated phosphoinositide hydrolysis in rat choroid plexus after fluoxetine and citalopram treatments. AB - Selective serotonin reuptake inhibitors (SSRIs) bind directly to various neurotransmitter receptors. The clinical effects of SSRIs appear gradually during weeks of treatment, suggesting a role for adaptive changes in neurotransmitter receptors. Most clinically used antidepressants, e.g. fluoxetine, bind to 5-HT2C receptors. When administered chronically, many antidepressants elicit adaptive regulation of 5-HT2C receptors. The present study was conducted in order to determine the effects of acute and chronic fluoxetine and citalopram treatments on the density and function of 5-HT2C receptors in the rat choroid plexus. Acute and chronic treatments followed by phosphoinositide (PI) hydrolysis assays and quantitative receptor autoradiography were performed. Acute (single-dose) treatment with neither drug significantly affected basal or 5-HT-stimulated PI hydrolysis, but acute citalopram (20 mg/kg) treatment increased both agonist and antagonist binding to 5-HT(2C) receptors. Chronic (14 days) citalopram treatment (20 mg/kg) increased the maximal PI hydrolysis response by 40%, but fluoxetine lacked this effect. The present data suggest that sensitisation of 5-HT2C receptor-mediated intracellular signal transduction may play a role in the effects of citalopram. In contrast, fluoxetine treatment does not functionally sensitise 5-HT2C receptors. Thus, functional 5-HT2C receptor sensitisation is not a common effect of antidepressants, but the differential effects may explain some of the pharmacodynamic differences seen with these drugs, especially upon repeated administration. PMID- 15749457 TI - Mangifera indica L. extract (Vimang) inhibits Fe2+-citrate-induced lipoperoxidation in isolated rat liver mitochondria. AB - The extract of Mangifera indica L. (Vimang) is able to prevent iron mediated mitochondrial damage by means of oxidation of reduced transition metals required for the production of superoxide and hydroxyl radicals and direct free radical scavenging activity. In this study we report for the first time the iron complexing ability of Vimang as a primary mechanism for protection of rat liver mitochondria against Fe2+ -citrate-induced lipoperoxidation. Thiobarbituric acid reactive substances (TBARS) and antimycin A-insensitive oxygen consumption were used as quantitative measures of lipoperoxidation. Vimang at 10 microM mangiferin concentration equivalent induced near-full protection against 50 microM Fe2+ citrate-induced mitochondrial swelling and loss of mitochondrial transmembrane potential (DeltaPsi). The IC50 value for Vimang protection against Fe2+ -citrate induced mitochondrial TBARS formation (7.89+/-1.19 microM) was around 10 times lower than that for tert-butylhydroperoxide mitochondrial induction of TBARS formation. The extract also inhibited the iron citrate induction of mitochondrial antimycin A-insensitive oxygen consumption, stimulated oxygen consumption due to Fe2+ autoxidation and prevented Fe3+ ascorbate reduction. The extracted polyphenolic compound, mainly mangiferin, could form a complex with Fe2+, accelerating Fe2+ oxidation and the formation of more stable Fe3+ -polyphenol complexes, unable to participate in Fenton-type reactions and lipoperoxidation propagation phase. The strong DPPH radical scavenging activity with an apparent IC50 of 2.45+/-0.08 microM suggests that besides its iron-complexing capacity, Vimang could also protect mitochondria from Fe2+ -citrate lipoperoxidation through direct free radical scavenging ability, mainly lipoperoxyl and alcoxyl radicals, acting as both a chain-breaking and iron-complexing antioxidant. These results are of pharmacological relevance since Vimang could be a potential candidate for antioxidant therapy in diseases related to abnormal intracellular iron distribution or iron overload. PMID- 15749458 TI - Induction of apoptosis and inhibition of telomerase activity by aqueous extract from Platycodon grandiflorum in human lung carcinoma cells. AB - The objective of the present study was to investigate the effect of aqueous extract from the root of Platycodon grandiflorum (AEPG) on the cell growth and apoptosis in human lung carcinoma cell line A549. Exposure of A549 cells to AEPG resulted in growth inhibition and induction of apoptosis in a dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometry analysis. This increase in apoptosis was associated with a decrease in Bcl-2 expression, an increase of Bax and an activation of caspase-3. AEPG treatment markedly inhibited the activity of telomerase in a dose-dependent fashion. Additionally, the expression of human telomerase reverse transcriptase (hTERT), a main determinant of the telomerase enzymatic activity, was progressively down-regulated by AEPG treatment. These findings suggest that the apoptotic events by AEPG were associated with the diminished telomerase activity and down-regulation of Bcl-2 expression. PMID- 15749459 TI - Assessment of sex differences in pharmacokinetics and pharmacodynamics of amlodipine in a bioequivalence study. AB - AIMS: This study was conducted to assess the bioequivalence between two 10-mg amlodipine tablet formulations. As secondary objectives, sex-related differences and tolerability profile were evaluated. METHODS: Thirty-six healthy volunteers (18 males and 18 females; age 20-32 years, weight 49.5-98.0 kg) were included in a randomised crossover study. Subjects were administered a single 10-mg oral dose of each formulation separated by a 14-day washout period. Plasma amlodipine levels were determined by a high performance liquid chromatographic method with tandem mass spectrometry detection. RESULTS: All subjects completed the study and 90% confidence intervals for relevant pharmacokinetic parameters were within the ranges defined by European and US Regulatory Authorities: the geometric mean and the 90% confidence interval test/reference ratios calculated from log-transformed values were 104.54 (101.46-107.72%) for AUC(0-infinity) and 100.32 (97.41 103.33%) for Cmax. There were no serious or severe adverse events. The tolerability profile appeared to be comparable for the two products. On average, bioavailability of amlodipine was slightly higher in females than in males, but these differences could be explained by the lower body weight of women. There were no sex-related differences in drug clearance. Bioequivalence was also demonstrated within each gender group. Amlodipine treatment produced a slight decrease of systolic blood pressure and an increased in heart rate, which were more pronounced in women. The incidence of adverse events was similar in men and women. CONCLUSIONS: The two formulations were considered bioequivalent. Although there were no relevant gender-related differences in the pharmacokinetics of amlodipine, women reached higher amlodipine concentrations most likely because of their lower body weight, and therefore, the reported pharmacodynamic effects were higher within this gender group. PMID- 15749460 TI - Inhibition of poly(ADP-ribose) polymerase attenuates lung tissue damage after hind limb ischemia-reperfusion in rats. AB - The objective of this study was to investigate the effects of 3-aminobenzamide (3 AB) on tissue damage in lung after hind limb ischemia-reperfusion (I/R), by assessing blood biochemical assay and histopathological analysis. Thirty-five adult Wistar rats were divided into five groups. After application of anaesthesia both hind limbs were occluded with tourniquets. Following ischemia period for 60 min, the tourniquets were removed allowing reperfusion for 120 min. The IR group received 0.5 ml of saline while the IR+AB group received 3-AB (10 mgkg(-1) intraperitoneally). The IR+DMSO group was given 0.5 ml 10% DMSO 30 min before the removal of the tourniquets. The control group received 0.5 ml saline and the AB group received 0.5 ml 3-AB (10 mgkg(-1)) intraperitoneally. At the end of the reperfusion period, mid-line sternotomy was performed. Blood samples were taken with cardiac puncture. Bronchoalveolar lavage (BAL) of the left lung was performed with saline. Right lung was preserved for histopathological evaluation and biochemical examination. Lung tissue malondialdehyde (MDA) and 3 nitrotyrosine levels, myeloperoxidase and Na+/K+ ATP-ase activities, wet to dry weight ratios, and plasma and BAL fluid MDA levels were determined. Histopathological evaluation was performed, too. Hind limb IR caused significant increase in the lung tissue 3-NT to total tyrosine ratio (p = 0.014), wet to dry weight ratio (p = 0.000), MPO activity (p = 0.000), and MDA levels (p = 0.000). The animals treated with 3-AB showed a statistically significant decrease in these values (p < 0.05). Na+/K+ ATP-ase activity which was found to be decreased significantly with IR, returned to near normal levels with 3-AB treatment. Additionally, lung tissue injury in IR group characterized with moderate interstitial congestion and neutrophil infiltration, showed remarkable amelioration following 3-AB treatment. Our results strongly support the view that poly(ADP-ribose) polymerase (PARP) plays an important role in the inflammatory process in hind limb I/R-induced lung injury and as a PARP inhibitor, 3-AB seems to have a potential to treat this inflammatory injury. PMID- 15749462 TI - The effect of hyaluronic acid on interleukin-1-induced deregulation of collagen metabolism in cultured human skin fibroblasts. AB - Although hyaluronic acid (HA) has been used in the treatment of osteoarthritis for 30 years, the mechanism of its protective action on collagen metabolism disturbances in tissues during inflammation is not known. The present study was undertaken to evaluate the mechanism of Interleukin-1 (IL-1)-induced deregulation of collagen metabolism in cultured human skin fibroblast and the effect of HA on the process. In normal fibroblasts IL-1 strongly induced inhibition of collagen biosynthesis, while HA counteracted the process. The mechanism of this phenomenon was independent of prolidase activity, an enzyme that plays an important role in collagen biosynthesis at the post-translational level. Instead, IL-1 was found to inhibit the expression of insulin-like growth factor-I receptor (IGF-IR) and MAP kinases-ERK1 and ERK2, while HA was shown to counteract this process. Since insulin-like growth factor-I (IGF-I) is a most potent stimulator of collagen biosynthesis in fibroblasts the mechanism of IL-1-dependent inhibition of collagen biosynthesis may be related to inhibition of IGF-IR expression and signaling. The data suggest that hyaluronic acid protects collagen against IL-1 induced inhibition of biosynthesis of this protein in cultured human skin fibroblasts at the level of IGF-IR signaling. PMID- 15749461 TI - Role of p38 MAP kinase in postcapillary venule leukocyte adhesion induced by ischemia/reperfusion injury. AB - Inflammation and leukocyte activation/infiltration play a major role in the initiation and progression of cardiovascular diseases including atherosclerosis and heart failure. Acute p38 mitogen-activated protein kinase (MAPK) pathway inhibition attenuates tissue damage and leukocyte accumulation in myocardial ischemia/reperfusion injury, although its effect on the acute phase of leukocyte recruitment has not been elucidated. The purpose of this study was to test the hypothesis that acute treatment of rats with a selective p38 inhibitor, SB 239063, inhibits ischemia/reperfusion-induced leukocyte-endothelial adhesion in vivo. Male Sprague-Dawley rats were treated with either SB-239063 (10 mgkg(-1)), dexamethasone (3 mgkg(-1)) or vehicle 1h prior to ischemia. Postcapillary venules were observed microscopically in exteriorized, superfused cremaster tissue. Leukocytes were fluorescently labeled in vivo using intravenous rhodamine 6G. Leukocyte adhesion, rolling, and rolling velocities were quantitated prior to 30 min ischemia, and at several time points during a 90 min reperfusion period. Ischemia caused a 3-fold increase in adherent leukocytes 5 min following reperfusion, a response that was maintained throughout the monitoring period (90 min) in vehicle-treated animals. SB-239063, at a dose known to inhibit p38 MAPK activity in vivo (10 mgkg(-1)), had no effect on ischemia/reperfusion-induced leukocyte adhesion, the number of rolling leukocytes, rolling velocities during the reperfusion period or adhesion molecule expression (P-, E-selectin, VCAM-1, ICAM-1). In contrast, dexamethasone completely blocked leukocyte adhesion in response to ischemia/reperfusion, and reduced expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). We conclude that p38 MAPK may not play a role in initial leukocyte recruitment in response to ischemia/reperfusion injury, but could affect leukocyte emigration, thereby resulting in increased leukocyte accumulation in ischemic-reperfused tissue. PMID- 15749463 TI - Protective effect of ipratropium bromide on bronchoconstriction induced by sulfur dioxide exposure during apricot sufurization processes that causes asthma-like syndrome in agricultural environment. AB - We previously showed that apricot sulfurization workers are exposed to high concentrations of sulfur dioxide (SO2), resulting in an asthma-like syndrome. The aim of this study was to investigate whether pre-treatment of ipratropium bromide protects bronchoconstriction induced by SO2 exposure during apricot sulfurization processes that causes asthma-like syndrome. Firstly, pulmonary function tests were measured before and immediately after SO2 exposure due to processes of apricot sulfurization in 21 healthy volunteer apricot sulfurization workers who did not use any medication in apricot farms. One week later, same measurements were repeated in the same workers when they were working in same farm but they were administered two puffs of ipratropium bromide (20 microg per dose) before 30 min second SO2 exposure for protection of SO2-induced bronchoconstriction. Occupational SO2 exposure caused significant decrement in forced vital capacity (FVC), forced expiratory volume (FEV1) and forced mid-expiratory flow rate (FEF25 75%) in the worker and these decrements were prevented by ipratropium bromide given 30 min before SO2 exposure. This result suggests that pre-treatment of ipratropium bromide protects SO2-induced bronchoconstriction in healthy worker during apricot sulfurization processes that causes asthma-like syndrome in agricultural environment. PMID- 15749464 TI - Blockade of the expression of mecamylamine-precipitated nicotine withdrawal by calcium channel antagonists. AB - The present study focused on the evaluation of nicotine abstinence syndrome in mice and on the influence of calcium channel blockers on the expression of the somatic signs of nicotine withdrawal. Our experimental protocol consisted of intermittent administration of nicotine, 2.5 mgkg(-1), subcutaneously (s.c.), four times daily for 7 days. In attempt to precipitate nicotine abstinence, mice were given one injection of mecamylamine (3 mgkg(-1), intraperitoneally (i.p.)), 1h after the last nicotine injection, on the test day (day 8) in the morning. Additionally, body weight changes, locomotor activity and anxiogenic responses in the elevated plus maze test were also evaluated in nicotine withdrawn mice. Our data shown that the L-type calcium channel antagonists, nimodipine, verapamil, flunarizine and diltiazem (5 and 10 mgkg(-1), i.p., each), injected before mecamylamine administration, dose-dependently attenuated the expression of nicotine withdrawal signs. Moreover, 24h after terminating nicotine treatment, we also observed additional nicotine abstinence measures, such as loss of body weight followed by a slight body weight gain, decrease of spontaneous locomotor activity and anxiogenic responses. These findings obtained using our valuable rodent model of nicotine dependence suggest the involvement of calcium-dependent mechanisms in the expression of mecamylamine-precipitated nicotine abstinence syndrome. PMID- 15749465 TI - Beneficial interaction between vigabatrin and valproate against seizures induced by pentylenetetrazole in mice. AB - The anticonvulsant effects of adding a non-protective dose of vigabatrin (VGB) to increasing single doses of sodium valproate (VPA) against seizures induced by 110 mgkg(-1) of pentylenetetrazole (PTZ) or by 4.5 mgkg(-1) of picrotoxin (PIC) were compared in CD1 mice with those of VPA alone and vice versa. Neurotoxicity was evaluated by the rotarod test. The study also assessed changes in concentrations of anticonvulsants, gamma-aminobutyric acid (GABA) and glutamate in the whole brain. VGB increased the potency ratio of VPA against PTZ (1.62, P < 0.05) but not against PIC (1.08, n.s.). VGB slightly decreased the neurotoxicity of VPA (0.93, n.s.) and the protective index of VPA was, therefore, increased from 1.93 to 3.34 for the PTZ model and from 1.40 to 1.61 for the PIC model. VGB did not modify brain concentrations of VPA, and increased brain GABA in relation to VPA alone. On the other hand, VGB did not achieve a complete protection neither against seizures induced by PTZ nor against seizures induced by PIC and a non protective dose of VPA did not significantly modify the effects of increasing doses of VGB. In conclusion, the addition of a non-protective dose of VGB increased the anticonvulsants effects and the protective index of VPA in the PTZ model. A more than expected brain GABA increase together with the lack of a pharmacokinetic interaction support a pharmacodynamic basis for this interaction. PMID- 15749466 TI - Modular cloning in plant cells. AB - New plant genes are being discovered at a rapid pace. Yet, in most cases, their precise function remains elusive. The recent advent of recombinational cloning techniques has significantly improved our ability to investigate gene functions systematically. For example, proteins fused with diverse fluorescent tags can be expressed at will using versatile cloning cassettes. In addition, novel binary T DNA vectors are now available to assemble multiple DNA fragments simultaneously, which greatly facilitate plant cell and protein engineering. PMID- 15749467 TI - Plant synapses: actin-based domains for cell-to-cell communication. AB - For many years it has been known that plants perform rapid long-distance signalling using classical action potentials that have impacts on diverse processes in plants. Plants also synthesize numerous neuronal molecules and fulfill some criteria for intelligent behaviour. Analysis of recent breakthrough data from ecophysiology studies has revealed that plant roots can discriminate between 'self' and 'non-self'; in animals, this ability to discriminate is dependent on the activities of neuronal synapses. Here, we propose that plant cells establish modes of information exchange between each other that have properties in common with neuronal synapses. Moreover, plants also assemble adhesive contacts that orchestrate cell-to-cell communication between the host cells when challenged with pathogens, parasites and potential symbionts. We propose that these adhesive contacts resemble the immunological synapses found in animals. PMID- 15749468 TI - Relief for fish stocks: oceanic fatty acids in transgenic oilseeds. AB - Three recent reports (Baoxiu Qi et al., Amine Abbadi et al. and Anthony J. Kinney et al.) describe the production of very long-chain polyunsaturated fatty acids in transgenic plants. This might lead to a sustainable source of these valuable fatty acids for use in human food and animal feed. At present they are mainly available via consumption of fish, which is a limited and endangered resource. PMID- 15749469 TI - Many ways to exit? Cell death categories in plants. AB - Programmed cell death (PCD) is an integral part of plant development and defence. It occurs at all stages of the life cycle, from fertilization of the ovule to death of the whole plant. Without it, tall trees would probably not be possible and plants would more easily succumb to invading microorganisms. Here, we have attempted to categorize plant PCD in relation to three established morphological types of metazoan cell death: apoptosis, autophagy and non-lysosomal PCD. We conclude that (i) no examples of plant PCD conform to the apoptotic type, (ii) many examples of PCD during plant development agree with the autophagic type, and (iii) that other examples are apparently neither apoptotic nor autophagic. PMID- 15749470 TI - Tall tales from sly dwarves: novel functions of gibberellins in plant development. AB - Gibberellins (GAs) are endogenous hormones controlling numerous aspects of plant growth and development. Our present understanding of GA physiology is based largely on genetic analysis in model plants such as Arabidopsis. In spite of the success of this approach, the discovery of additional physiological roles for GAs in seed development, pollen tube growth and meristem development indicates that the existing collection of GA-related mutants (identified partially or entirely on the basis of vegetative phenotypes) has failed to uncover all aspects of plant development that are controlled by GAs. The continued use of ever improving forward and reverse genetic techniques is expected to lead to the discovery of further novel roles for GAs in plant development. PMID- 15749471 TI - Leaf starch degradation comes out of the shadows. AB - During the day, plants accumulate starch in their leaves as an energy source for the coming night. Based on recent findings, the prevailing view of how the transitory starch is remobilized needs considerable revision. Analyses of transgenic and mutant plants demonstrate that plastidic glucan phosphorylase is not required for normal starch breakdown and cast doubt on the presumed essential role of alpha-amylase but do show that beta-amylase is important. Repression of the activity of a plastidic beta-amylase, the export of its product (maltose) or further metabolism of maltose by a newly identified transglucosidase impairs starch degradation. Breakdown of particulate starch also depends on the activity of glucan-water dikinase, which phosphorylates glucosyl residues within the polymer. PMID- 15749472 TI - Cereal phytochromes: targets of selection, targets for manipulation? AB - Plants respond to shading through an adaptive syndrome termed shade avoidance. In high-density crop plantings, shade avoidance generally increases extension growth at the expense of yield and can be at odds with the agronomic performance of the crop as a whole. Studies in Arabidopsis are beginning to reveal the essential role phytochromes play in regulating this process and to identify genes underlying the response. In this article, we focus on how phytochrome signaling networks have been targeted in cereal breeding programs in the past and discuss the potential to alter these pathways through breeding and transgenic manipulation to develop crops that perform better under typical high density conditions. PMID- 15749473 TI - Geminiviruses and RNA silencing. AB - Geminiviruses are single-stranded circular DNA viruses that cause economically significant diseases in a wide range of crop plants worldwide. In plants, post transcriptional gene silencing (PTGS) acts as a natural anti-viral defense system and plays a role in genome maintenance and development. During the past decade there has been considerable evidence of PTGS suppression by viruses, which is often required to establish infection in plants. In particular, nuclear replicating geminiviruses, which have no double-stranded RNA phase in their replication cycle, can induce and suppress the PTGS and become targets for PTGS. Here, we summarize recent developments in determining how these viruses trigger PTGS and how they suppress the induced PTGS, as well as how we can use the system to control these viruses in plants better and manipulate the system to study functional genomics in crop plants. PMID- 15749474 TI - Reduced progesterone action during endometrial maturation: a potential risk factor for the development of endometriosis. AB - OBJECTIVE: To discuss the role that reduced endometrial responsiveness to progesterone (P) might play in the pathophysiology of endometriosis. DESIGN: A review of experimental evidence regarding the failure of P to regulate the expression of matrix metalloproteinases (MMPs) in the endometrium of patients with endometriosis. CONCLUSION(S): Progesterone and locally produced differentiation factors act cooperatively to reduce MMP expression by maternal endometrial cells within the pro-inflammatory micro-environment of early pregnancy. Our in vitro studies with normal human endometrium demonstrate that prior P exposure not only down-regulates MMP expression, but also limits the ability of locally produced proinflammatory cytokines to stimulate expression of these enzymes. In contrast, endometrial tissues from women with endometriosis demonstrate an altered response to P, allowing a continuous expression of MMPs throughout the secretory phase. Although the factors that influence the loss of P sensitivity in the endometrium of patients with endometriosis have not yet been defined, alterations in cell-cell communication seem to contribute to dysregulated MMP expression. Specifically, proinflammatory cytokines produced by epithelial cells oppose stromal cell responses to P, inhibiting production of key differentiation factors necessary for cell-specific MMP regulation. The resulting loss in normal MMP regulation enhances the invasive capacity of endometrial tissue, promoting ectopic establishment in an experimental model. PMID- 15749475 TI - Quality of fertility clinic websites. AB - OBJECTIVE: To evaluate the overall quality of the Society for Assisted Reproductive Technology (SART)-affiliated fertility clinic websites, according to American Medical Association (AMA) Internet health information guidelines. DESIGN: Cross-sectional evaluation. SETTING: Two hundred sixty-six websites drawn from the SART directory and the World Wide Web. INTERVENTION(S): Three objective scoring scales (ownership, content, and navigation) adapted from AMA guidelines. MAIN OUTCOME MEASURES: Seven objective criteria for ownership, 8 for content, and 11 for website navigation. RESULT(S): Two thirds of SART-affiliated fertility clinics have functional websites. Of the 236 sites evaluated, 58 belong to hospital centers. Overall, the scores for the three scoring scales were low. Compared with the websites of non-hospital clinics, those of hospital centers were more likely to include information about site ownership and affiliations (89.7% vs. 60.7%) and patient privacy (34.5% vs. 20.8%). Also, contents of hospital center websites were significantly easier to distinguish from advertisements (70.7% vs. 47.7%), and reference sources for specific contents were easier to identify (27.6% vs. 8.4%). Hospital center websites were more likely to indicate affiliations and financial disclosures of authors (25.9% vs. 10.7%), to feature a site map (25.9% vs. 12.3%), and to have a search function (31.0% vs. 5.6%). CONCLUSION(S): Websites of SART-affiliated clinics fail to meet most of the AMA health information guidelines. The quality of the hospital centers' websites is better than that of private clinics. PMID- 15749476 TI - Website quality assessment: mistaking apples for oranges. AB - Fertility clinic websites are primarily a form of advertising. The American Medical Association Internet health information guidelines were not designed to assess the quality of such websites. PMID- 15749477 TI - Veracity in in vitro fertilization Web pages. AB - Huang et al. described compliance of IVF websites against the American Medical Association online health information guidelines and reported that IVF websites scored poorly. We describe a protocol for IVF websites that would inform readers about truthfulness of the page, develop standards for page construction, and establish a review process. PMID- 15749478 TI - Assisted reproductive technologies on the Web. AB - The rapid advance of the World Wide Web presents significant challenges for ensuring quality of health care information presented to a global audience. Using American Medical Association guidelines, Huang and co-authors have issued a report card for Society for Assisted Reproductive Technology-affiliated websites, indicating much room for improvement. PMID- 15749479 TI - Assessing infertility information on the Internet: challenges and possible solutions. AB - Although patients frequently visit consumer-related Internet sites to help them make informed decisions about treatment, there is no clear way for these consumers to evaluate content within these sites. This article reviews the problem and offers suggestions to remedy the situation. PMID- 15749480 TI - A call for standardization of fertility clinic websites. AB - Websites for Society for Assisted Reproductive Technology-affiliated clinics fail to meet most American Medical Association health information guidelines. Professional organizations in reproductive medicine need to standardize the accuracy and appropriateness of online reproductive health information. PMID- 15749481 TI - Postmenopausal hormone therapy: critical reappraisal and a unified hypothesis. AB - OBJECTIVE: To reconcile apparently conflicting evidence regarding the use of hormone therapy as a health-preserving strategy in postmenopausal women in light of that fact that findings from animal studies, human observation studies, and human clinical trials are consistent for outcomes such as fracture and breast cancer but differ for coronary heart disease (CHD). DESIGN: Literature review and generation of a unified hypothesis consistent with all of the data. SETTING: Animal trials, human observational studies, human studies of biologic intermediates, and human clinical trials. PATIENT(S): Premenopausal and postmenopausal women with or without antecedent CHD. MAIN OUTCOME MEASURE(S): Coronary heart disease events, proxies, risk factors, and related mechanisms. RESULT(S): The complex CHD responses to hormone therapy in recent human trials likely reflect a combination of [1] early erosion/rupture of "vulnerable" coronary plaque, which is made worse by hormone therapy, [2] long-term reduction in plaque formation, which is improved by hormone therapy, and [3] modulation of the vasculoprotective actions of estrogens by systemic progestogens. CONCLUSION(S): The unified hypothesis predicts that hormone therapy initiated at the time of menopause should produce a decrease in CHD over time. In contrast, hormone therapy begun years after menopause should produce an increase in CHD events shortly after therapy is begun, followed later by benefit. In women who require progestogens for endometrial protection, there should be greater CHD benefit from use of progestogens with less systemic activity. The unified hypothesis is consistent both with plausible biologic mechanisms and with evidence from animal studies, human observational studies, and human clinical trials such as the Women's Health Initiative. In the absence of evidence from human trials that specifically involve initiation of hormone therapy in perimenopausal women, practitioners and patients can use the unified hypothesis as a rational tool to guide decisions about clinical management. PMID- 15749482 TI - Estrogen receptor alpha dinucleotide repeat and cytochrome P450c17alpha gene polymorphisms are associated with susceptibility to endometriosis. AB - OBJECTIVE: To investigate the association of endometriosis with estrogen receptor alpha (ER alpha) and cytochrome P450c17alpha (CYP17) gene polymorphisms in light of the fact that estrogen plays a role in the pathogenesis of endometriosis and the CYP17 enzyme is involved with estrogen biosynthesis. DESIGN: Prospective study. SETTING: Genetics and gynecology units. PATIENT(S): All patients were divided into two groups: group 1, women with endometriosis (n = 119); group 2, normal controls (n = 108). INTERVENTION(S): A dinucleotide (thymine-adenine [TA]) repeat polymorphism lying upstream of the ER alpha gene and A1/A2 polymorphism of the CYP17 gene were amplified by polymerase chain reaction, enzyme restriction, and electrophoresis. MAIN OUTCOME MEASURE(S): The ER genotypes were classified into A through T (TA repeats, 10-29). The CYP17 genotypes included indigestible (A1 homozygote), heterozygote, and digestible (A2 homozygote). We compared these polymorphism distributions in both groups. RESULT(S): The percentage of genotypes D-G (TA, 13-16) in both groups were 10.5%, 29.4%, 13.0%, and 11.3% in group 1 and 7.9%, 16.7%, 19.9%, and 17.6% in group 2. The genotype E (14 TA repeats) is associated with a higher risk of endometriosis. Proportions of A1 homozygote/heterozygote/A2 homozygote for CYP17 were 26.1%/46.2%/27.7% for group 1 and 14.8%/44.5%/40.7% for group 2, respectively. The A1 homozygote and allele were associated with a higher susceptibility of endometriosis. CONCLUSION(S): ER alpha* 14 TA repeats and the CYP17* A1 allele are associated with an increased risk of endometriosis. Both polymorphisms are useful markers for predicting endometriosis susceptibility. PMID- 15749483 TI - Quality of sex life in women with endometriosis and deep dyspareunia. AB - OBJECTIVE: To characterize sexual function among women with endometriosis and deep dyspareunia (DD). DESIGN: Cross-sectional survey. SETTING: University teaching hospital. PATIENT(S): Three-hundred nine women undergoing surgery because of infertility, pelvic pain, or adnexal masses. Three groups of patients with DD were created: women with deep infiltrating endometriosis of the uterosacral ligament (group U), women with endometriosis without uterosacral ligament lesions (group E), and controls (group C). INTERVENTION(S): Laparoscopy. MAIN OUTCOME MEASURE(S): Sexual function questionnaire. RESULT(S): The prevalence of DD since the first intercourse was significantly higher among women with endometriosis than in controls (P=.029). When group U was compared with group E and C, the pain score was higher, the number of intercourses per week was reduced, the orgasm was less satisfying, and the patients felt less relaxed and fulfilled after sex. No significant difference was observed in pain score and coital frequency between subjects with monolateral and bilateral lesions of the uterosacral ligament. CONCLUSION(S): Among subjects with DD, those with deep infiltrating endometriosis of the uterosacral ligament have the most severe impairment of sexual function; the presence of bilateral lesions does not influence the severity of the symptoms. Women with endometriosis have frequently suffered DD during their entire sex lives. PMID- 15749485 TI - Differential expression of endometrial integrins and progesterone receptor during the window of implantation in normo-ovulatory women treated with clomiphene citrate. AB - OBJECTIVE: To assess the effect of clomiphene citrate (CC) on endometrial epithelial integrins and P receptors (PR) during the window of implantation. DESIGN: Controlled, prospective, clinical study. SETTING: Teaching hospital and university research laboratory. PATIENT(S): Thirty-one fertile, normo-ovulatory women participated in this trial. Thirteen women exhibited a CC-stimulated cycle with 50 mg on days 5-9, and 18 women with spontaneous menstrual cycles served as controls. INTERVENTION(S): Endometrial biopsies in the midluteal phase. MAIN OUTCOME MEASURE(S): Immunohistochemical determination and endometrial cellular localization of alpha1, alpha v, beta3, and alpha4 epithelial integrins and PR during the window of implantation. The staining intensity was assessed by a semiquantitative index (HSCORE) and compared by nonparametric Mann-Whitney test. RESULT(S): Higher plasma levels of P and E2 and delayed histologic dating of the endometrium (38%) were features of CC-treated women. In addition, a low epithelial beta3 integrin expression and persistent PR were observed in glandular epithelial cells of "out-of-phase" endometrial biopsies from CC-treated women. In contrast, in "in-phase" biopsies, neither epithelial PR nor beta3 integrin were different from spontaneous control cycles. There was no difference in the expression of alpha1, alpha v, and alpha4 between the groups studied. CONCLUSION(S): The administration of clomiphene produces aberrant endometrial beta3 integrin expression in conjunction with a failure in the down-regulation of PR during the window of implantation in a significant number of normo-ovulatory women, notwithstanding the higher plasma P levels. Therefore, CC might affect the expression of endometrial receptivity markers. PMID- 15749484 TI - Heparanase improves mouse embryo implantation. AB - OBJECTIVE: To improve mouse embryonic implantation by recombinant heparanase supplementation. Heparanase, an endoglycosidase-degrading heparan sulfate proteoglycan, may have a role in embryonic implantation because of its enzymatic, angiogenic, and adhesive properties. Increasing endometrial receptivity could improve one of the most difficult pathologies in human fertility. DESIGN: Comparison between mouse blastocysts obtained after 24-hour incubation of morulae with or without heparanase. SETTING: Experimental laboratory in a medical center. ANIMAL(S): Mice. INTERVENTION(S): Morulae were flushed from CB6F1 female mice and incubated for 24 hours at 37 degrees C in M16 medium supplemented with 0.1 mg/mL heparanase (n = 203), with albumin (n = 60), or with medium alone (n = 258). MAIN OUTCOME MEASURE(S): Blastocysts were evaluated by heparanase immunostaining (n = 10), activity assay (n = 283), and transfer to foster mice uterine horns (n = 228). The number of implantation sites was compared. RESULT(S): Immunostaining demonstrated that heparanase is constitutively expressed in mouse morulae and blastocyts. Heparanase supplementation resulted in increased staining and enzymatic activity in blastocyts. Implantation rates for the heparanase, M16 medium, and albumin groups, were 36.9%, 17.8%, and 20%, respectively (P<.01). CONCLUSION(S): Heparanase was found to be constitutively expressed by blastocyst stage embryos. Moreover, the amount of heparanase was markedly increased by incubation of morulae with recombinant heparanase, evaluated by immunostaining and enzymatic activity. Heparanase supplementation resulted in approximately a twofold increase in embryo implantation rate in vivo. Taken together, these data suggest that heparanase is actively involved in embryo implantation. PMID- 15749486 TI - Immediate ambulation after embryo transfer: a prospective study. AB - OBJECTIVE: To assess whether bed rest following the embryo transfer (ET) procedure contributes to the implantation process and pregnancy rate. DESIGN: A prospective (patient-influenced) study. SETTING: An in vitro fertilization (IVF) unit of an academic medical center. PATIENT(S): Four hundred six patients undergoing controlled ovarian hyperstimulation and IVF. INTERVENTION(S): All women undergoing in vitro fertilization-embryo transfer (IVF-ET) cycles in our unit were given a special individual counseling session before the ET procedure in which they were informed that our previous experience showed no advantage for bed rest over immediate ambulation after ET. The women were allowed to select the practice of their choice, and they were assured that their decision would have no influence on their further treatment. MAIN OUTCOME MEASURE(S): The stimulation pattern and cycle outcome were compared between the two groups (bed rest and immediate ambulation). RESULT(S): Of the 406 patients counseled during the study period, 167 preferred immediate ambulation and 239 opted to stay in the unit for 1 hour's bed rest. There were no significant differences between the groups in mean patient age, number of embryos transferred, and other variables of the assisted reproductive technique cycles. Pregnancy rates did not differ between the groups: 41 out of 167 (24.55%) in the immediate-ambulation group and 51 out of 239 (21.34%) in the bed-rest group. CONCLUSION(S): Immediate ambulation following the ET procedure has no adverse influence on the ability to conceive. PMID- 15749487 TI - Role of the endometrial tripod interleukin-18, -15, and -12 in inadequate uterine receptivity in patients with a history of repeated in vitro fertilization-embryo transfer failure. AB - OBJECTIVE: To document in the endometrium the correlation among the interleukin (IL)-12, -15, and -18 mRNA and the correlation between cytokine levels, vascular status, and endometrial natural killer (NK) cell count in the context of recurrent implantation failure. DESIGN: A pilot study. SETTING: Department of Reproductive Immunology. PATIENT(S): Women who failed to become pregnant after repeated IVF-embryo transfer and fertile control subjects. INTERVENTION(S): Ultrasonic evaluation and endometrial biopsy in luteal phase. MAIN OUTCOME MEASURE(S): Uterine artery Doppler, count of uterine CD56 bright cells/field, and quantification by real-time polymerase chain reaction (PCR) to monitor IL-12 family (IL-12p40, IL-12p35, EBI3, IL-23), the IL-18 system (IL-18, IL-18R, IL18BP), and the IL-15 mRNA ratio. RESULT(S): The uterine artery Doppler and the CD56 bright cell counts were significantly different in fertile and infertile patients. The mean uterine artery pulsatility index correlated significantly negatively with the IL-18/actin ratio suggesting a defect of the cytokine dependent vascular remodeling pathway. The number of uterine CD56 bright cells was significantly correlated with the IL-15/actin and IL-18/IL-18BP ratios. Thus, IL-18 and IL-15 seems to be involved in the local recruitment and the activation of uterine natural killer (uNK) cells. IL-18 was itself correlated with IL-15 and IL-12, suggesting a local control of uNK cells activation. CONCLUSION(S): The assessment of the tripod IL-12/-15/-18 shows distinct immune-related mechanisms that are involved in the broader context of inadequate uterine receptivity. PMID- 15749488 TI - Etiology-specific outcomes of intracytoplasmic sperm injection in azoospermic patients. AB - OBJECTIVE: To assess fertilization, pregnancy, and miscarriage rates after intracytoplasmic sperm injection (ICSI) with epididymal or testicular spermatozoa from different types of azoospermia. DESIGN: Retrospective study. SETTING: Academic medical center and private fertility center. PATIENT(S): Two hundred twelve patients underwent 257 ICSI cycles. INTERVENTION(S): Cycles of ICSI were divided into four groups according to the etiology of azoospermia: A (nonobstruction), B (postvasectomy), C (congenital obstruction), and D (obstruction due to infection). Testicular sperm aspiration and percutaneous epididymal sperm aspiration were the sperm retrieval methods used for ICSI. MAIN OUTCOME MEASURE(S): Fertilization, pregnancy, and miscarriage rates. RESULT(S): Normal fertilization rates were higher in groups C (67.7%) and B (64.1%) compared with groups A (47.3%) and D (58.9%). Although lower pregnancy rates were seen in group A, no statistical differences were detected among groups. However, the miscarriage rate was higher in group A (45.6%) compared with groups B (25.25%), C (24%), and D (22.58%). CONCLUSION(S): Although no differences were detected in the pregnancy rates across groups, fertilization and implantation rates were higher in patients with congenital obstruction of the seminal path. The pregnancy rate was higher and the miscarriage rate lower when epididymal sperm was used compared with testicular sperm. PMID- 15749489 TI - Intracytoplasmic sperm injection as a treatment for unexplained total fertilization failure or low fertilization after conventional in vitro fertilization. AB - OBJECTIVE: To determine whether IVF or intracytoplasmic sperm injection (ICSI) should be the choice of treatment in case of a previous IVF attempt with unexplained total fertilization failure or low fertilization (<25%). DESIGN: Prospective study. SETTING: Leiden University Medical Center. PATIENT(S): Thirty eight couples undergoing IVF and ICSI on sibling oocytes after a first IVF attempt with total fertilization failure or with low fertilization (<25%). INTERVENTION(S): Performing IVF and ICSI on sibling oocytes. MAIN OUTCOME MEASURE(S): Fertilization and (ongoing) pregnancy rate. RESULT(S): A total of 271 oocytes were collected in 24 oocyte retrievals in the total fertilization failure group. Hundred nine oocytes were randomly allocated to IVF and 12 were fertilized (11%); 162 sibling oocytes were allocated to ICSI and 78 were fertilized (48%). In 8 of the 24 patients fertilization occurred after IVF. The pregnancy rate after transfer of 1 IVF and 1 ICSI embryo (n = 3) was 67% and after the transfer of 2 ICSI embryos (n = 21) this was 52%. In the low fertilization group 169 oocytes were collected in 14 oocyte retrievals. Seventy-two oocytes were randomly allocated to IVF and 16 were fertilized (22%). Ninety-seven sibling oocytes were allocated to ICSI and 58 were fertilized (60%). In 7 of 14 patients fertilization occurred after IVF. The pregnancy rate after the transfer of 1 IVF and 1 ICSI embryo (n = 5) was 80% and after the transfer of 2 ICSI embryos (n = 9) this was 33%. CONCLUSION(S): Performing ICSI on some oocytes of a cohort may avoid total fertilization failures both in patients with a history of total fertilization failure and in patients with a history of low fertilization, as the percentage of fertilization is higher after ICSI compared to IVF and the recurrence of total fertilization failure and low fertilization is high after IVF treatment. PMID- 15749490 TI - Effect of sperm treatment with exogenous platelet-activating factor on the outcome of intrauterine insemination. AB - OBJECTIVE: To evaluate the effect of sperm treatment with exogenous platelet activating factor (PAF) on IUI clinical pregnancy rate. DESIGN: Prospective randomized study. SETTING: Assisted Reproduction Unit, 2nd Department of Obstetrics and Gynecology, University of Athens, Aretaieion Hospital, Athens, Greece. SUBJECT(S): Fifty-two couples with unexplained infertility, candidates for IUI. INTERVENTION(S): Sperm treatment with an exogenous mixture of PAF (final concentration, 10(-7) mol/L) in sperm-washing medium, direct swim-up technique of sperm preparation, a maximum of six IUI cycles per couple with or without PAF treatment. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (pregnancies confirmed by ultrasonography per hundred cycles). RESULT(S): The overall clinical pregnancy rate after a maximum of six IUI cycles was significantly higher when sperm was treated with PAF compared with the rate after the direct swim-up technique (23.07% vs. 7.92%). CONCLUSION(S): Treatment of sperm with exogenous PAF might improve the clinical outcome of IUI in cases of unexplained infertility. PMID- 15749492 TI - Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. AB - OBJECTIVE: To investigate whether there is an impact of different sources of reactive oxygen species (ROS) on sperm functions. DESIGN: Prospective study. SETTING: Patients at the Center for Dermatology and Andrology, Giessen, Germany. PATIENT(S): Semen collected from 63 randomly collected patients attending the IVF unit of the University of Giessen, Germany. INTERVENTION(S): Only patients with nonleukocytospermia were included in this study. MAIN OUTCOME MEASURE(S): Sperm count and motility before and after sperm separation by swim-up, morphology, DNA fragmentation, and extrinsic (by leukocytes) and intrinsic ROS production (by spermatozoa) were evaluated. RESULT(S): Leukocytes correlated significantly with extrinsic ROS production (r = 0.576), but markedly less with intrinsic ROS production (r = 0.296). Sperm count, morphology, and motility in the ejaculate were markedly more affected by extrinsic than by intrinsic ROS. The DNA fragmentation was strongly positively correlated with intrinsic ROS production, whereas this correlation was weaker for extrinsic ROS production. No correlation was found between DNA fragmentation and the number of leukocytes, whereas the correlations with motility in the ejaculate and the motile sperm count after swim up were highly significant. Moreover, significant differences were observed for extrinsic and intrinsic ROS production between groups of patients having a high (> or = 1 x 10(6)/mL) and a low number (<1 x 10(6)/mL) of leukocytes in the ejaculate. CONCLUSION(S): The origin of ROS seems to have an influence on the site of the damage. Because leukocyte counts <1 x 10(6)/mL caused a significant decrease of motility and DNA integrity, the threshold given by the World Health Organization (WHO) should be re-evaluated. PMID- 15749491 TI - Deletions in L-type calcium channel alpha1 subunit testicular transcripts correlate with testicular cadmium and apoptosis in infertile men with varicoceles. AB - OBJECTIVE: To identify and understand predictors of successful varicocelectomy. DESIGN: Examination of testicular L-type voltage-dependent calcium channel (L VDCC) mRNAs and proteins in testis biopsies and comparison of presence and absence of various mRNAs with testicular cadmium levels, with apoptosis, and with sperm count change after varicocelectomy. SETTING: University clinical urology practice and research laboratories. PATIENT(S): Infertile men with varicocele (left varicocele only, n = 18; bilateral varicoceles, n = 26) and controls (men with obstructive azoospermia undergoing testicular sperm extraction before intracytoplasmic sperm injection; n = 7). INTERVENTION(S): Left testis biopsies by percutaneous needle aspiration biopsy. Varicocele repair by subinguinal approach. MAIN OUTCOME MEASURE(S): Calcium channel mRNA sequence by reverse transcription-polymerase chain reaction and amplicon analysis; calcium channel protein distribution by immunocytochemistry; cadmium levels by atomic absorption and apoptosis by deoxynucleotidyl transferase labeling; and sperm counts in the ejaculate before and after varicocelectomy. RESULT(S): Calcium channel mRNAs are polymorphic in human testis biopsies from different men. Proteins from sequence deleted exons 7 and/or 8 localize to germ cell membranes. Expression of undeleted L-type calcium channel mRNAs correlates with normal testes cadmium and increased sperm count after varicocelectomy. Apoptosis is lower in such cases. CONCLUSION(S): Expression of normal testicular L-VDCC sequence in exons 7-8 predicts postvaricocelectomy sperm count increase. Deletions may alter calcium channel function and affect testicular cadmium and apoptosis. PMID- 15749493 TI - A longitudinal study of the effects of free testosterone and other psychosocial variables on sexual function during the natural traverse of menopause. AB - OBJECTIVE: This study examines the effect of free T levels on sexual function during the natural traverse of menopause. Other psychosocial variables, which may also contribute to change in sexual function, are studied-including participant's job satisfaction, satisfaction with financial resources, confidence in ability to manage symptoms, stressful life events, exercise, body image, and quality of personal relationships. DESIGN: Prospective clinical study. SETTING: Clinical research center, university hospital. PATIENT(S): Fifty-seven women between the ages of 45 and 55 years were enrolled at the beginning of the study. Thirty-seven women completed all but the final measurement of free T and 23 completed all aspects of the study at year 1 and year 5. INTERVENTION(S): Questionnaires were administered and blood samples obtained in year 1 and year 5. MAIN OUTCOME MEASURE(S): Free T, sexual satisfaction, and other psychosocial variables. RESULT(S): Exercise is the only variable significantly associated with sexual satisfaction. There was a lack of association between free T and sexual satisfaction both at year 1 and year 5. CONCLUSION(S): Focus on the hormonal aspects of menopause has promoted a disregard of other important psychosocial factors affecting sexual function. This study shows no correlation between T levels and sexual function, whereas exercise is clearly associated with sexual satisfaction. A key area for future research is the effect of lifestyle changes on sexual function in menopausal women. PMID- 15749494 TI - Preimplantation sex selection demand and preferences in an infertility population. AB - OBJECTIVE: To determine the demand and preferences of infertility patients for sex selection and the method and sex they would choose, and to investigate the relationship between these choices and their demographic and socioeconomic characteristics. DESIGN: Cross-sectional survey. SETTING: University hospital based fertility center. PATIENT(S): One thousand five hundred consecutive women who presented for infertility care. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Self-report questionnaire assessing the demand and preferences for sex selection. RESULT(S): Of respondents, 40.8% wanted to select the sex of their next child for no added cost. Of these patients, 45.9% had no living children and 48.4% had children all of one sex. After adjustment for observed predictors of gender preference, we found a significant preference for a female child among women who were older, not religious, willing to pay for sex selection, had more living children, had only sons, or had a diagnosis of male infertility. Nulliparous women did not significantly prefer one sex over the other. Among parous women, those with only daughters significantly desired to select a male child, whereas those with sons significantly desired to select a female child. In terms of the method of sex selection, 55.0%, 41.0%, and 4.0% of the patients would use sperm separation, preimplantation genetic diagnosis, or neither method, respectively. CONCLUSION(S): There is significant demand among infertility patients for preimplantation sex selection, with a significant portion of this demand coming from patients who do not have any children or have children all of one sex. PMID- 15749495 TI - Histological characteristics and altered expression of interleukins (IL) IL-13 and IL-15 in endometria of levonorgestrel users with different uterine bleeding patterns. AB - OBJECTIVE: To determine the relationship between uterine bleeding patterns in levonorgestrel users with endometrial histology and expression of interleukins (IL) IL-13 and IL-15. DESIGN: Prospective observational study. SETTING: Academic research center. PATIENT(S): Questionnaires were sent to patients (n = 578) who had levonorgestrel implants concerning bleeding patterns; 35 of these patients were identified to have regular cycle (n = 13), amenorrhea (n = 8), or metrorrhagia (n = 14). INTERVENTION(S): Endometrial biopsies, serum, histology, and immunostaining. MAIN OUTCOME MEASURE(S): Endometrial histological assessment and immunostaining for IL-13 and IL-15 and for blood levonorgestrel, E2, and progesterone levels by ELISA or RIA. RESULT(S): No correlation was found between circulating levonorgestrel, E2, or progesterone levels with the bleeding patterns, although a trend toward a lower E2 level was observed in patients with amenorrhea who had inactive endometrium. There was a direct correlation between bleeding patterns and endometrial histology, as well as IL-13 and IL-15 expression in patients with regular cycles and metrorrhagia, demonstrating secretory and proliferative endometrium, respectively. Some patients in each group were also identified as demonstrating endometritis. CONCLUSION(S): Endometrial histology may assist directing therapy and subsequently increasing compliance in progestin-only contraceptive users with irregular bleeding who fail to respond to standard therapies. Altered endometrial expression of IL-13 and IL 15, key cytokines in inflammatory and immune cell trafficking, may influence events, leading to irregular bleeding. PMID- 15749496 TI - Ganirelix acetate use in normal- and poor-prognosis patients and the impact of estradiol patterns. AB - OBJECTIVE: To evaluate cycle outcomes in patients with either poor or normal prognosis undergoing IVF treatment with a GnRH antagonist (ganirelix acetate) for LH suppression. DESIGN: Nonrandomized, noncontrolled, retrospective review. PATIENT(S): 204 patients, aged 23-41 years, undergoing IVF. INTERVENTION(S): Patients completed 225 consecutive cycles of IVF with a GnRH antagonist (Antagon; Organon, Roseland, NJ) for LH surge prevention. Sixty cycles were conducted in patients with a known poor prognosis, whereas 165 were conducted in patients with a normal IVF prognosis. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR), for the series as a whole and according to prognosis, and serum E2 patterns. RESULT(S): The PR per initiated cycle for the series as a whole was 33.3%. The pregnancy rate was 42.1% per ET for the entire series, with a cycle cancellation rate of 21%. When evaluated by prognosis, the poor-prognosis patients had PRs of 8.3% per attempt and 15% per transfer, whereas the normal-prognosis patients had PRs of 40% per attempt and 45% per transfer. Pregnancy rate did not vary by E2 pattern (drop, plateau, or rise). Oral contraceptive pretreatment was noted to be associated with high cancellation rates in the group of known poor responders, whereas for the group as a whole, cycle outcome was unaffected by the use of oral contraceptives. CONCLUSION(S): Use of GnRH antagonists in patients with an a priori poor IVF prognosis results in predictably poor outcomes. Patients without factors predicting poor outcome have acceptable PRs. The pattern of E2 rise immediately after initiation of GnRH antagonists does not predict cycle outcome. Oral contraceptives can be successfully used to schedule antagonist-based IVF cycles but might increase the risk of cycle cancellation in some patient populations. PMID- 15749497 TI - Risk factors for high-order multiple pregnancy and multiple birth after controlled ovarian hyperstimulation: results of 4,062 intrauterine insemination cycles. AB - OBJECTIVE: To determine factors responsible for high-order multiple pregnancy (HOMP) and high-order multiple births when multiple cycles of controlled ovarian hyperstimulation-IUI (COH-IUI) are performed. DESIGN: Retrospective analysis. SETTING: Private infertility clinic. PATIENT(S): Women (n = 2,272) who underwent 4,067 consecutive COH-IUI cycles. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): High-order multiple pregnancy rate, pregnancy rate (PR), and birth rate (PR) per cycle. RESULT(S): High-order multiple pregnancy was related to number of follicles of diameter > or = 10 mm, age, and treatment cycle. For age <32 years, HOMP was 6% for three to six follicles and 20% for seven or more follicles. For ages 32 to 37 years, HOMP was 5% for three to six follicles and 12% for seven or more follicles. In the first COH-IUI cycle, HOMP was 8% for three to six follicles and 15% for seven or more follicles. In the second cycle, HOMP did not occur unless there were more than six follicles. No HOMP occurred after the second cycle. Pregnancy rate did not increase significantly when there were more than four follicles. Continuing COH-IUI past the third cycle resulted in additional pregnancies in patients with one to eight follicles. CONCLUSION(S): High-order multiple pregnancy can be predicted by age and number of follicles of diameter > or = 10 mm. Controlled ovarian hyperstimulation is not necessary to achieve satisfactory overall pregnancy rates if ovulation induction is continued past the third cycle in low responders. PMID- 15749498 TI - Antiphospholipid antibodies associated with recurrent pregnancy loss: prospective, multicenter, controlled pilot study comparing treatment with low molecular-weight heparin versus unfractionated heparin. AB - OBJECTIVE: To evaluate the use of low-molecular-weight heparin (LMWH) in combination with low-dose aspirin (LDA) for the treatment of antiphospholipid antibody (APA)-associated recurrent pregnancy loss and to compare the results with the use of unfractionated heparin (UFH) plus LDA. DESIGN: Prospective, controlled, multicenter pilot study. SETTING: Two academically based reproductive health centers. PATIENT(S): Patients with three or more pregnancy losses and positive APA. INTERVENTION(S): Patients were treated with LMWH and LDA (n = 25) or UFH and LDA (n = 25). MAIN OUTCOME MEASURE(S): Fetal outcome and maternal complications from treatments were compared between the two treatment groups. RESULT(S): Of the 25 patients in the LMWH group, 21 (84%) delivered a viable infant and 4 (16%) miscarried. Of the 25 patients in the UFH group, 20 (80%) delivered a viable infant and 5 (20%) miscarried. These differences were not statistically significant. No major bleeding episodes occurred during pregnancy or at the time of delivery. No cases of deep venous thrombosis, thrombocytopenia, pre-eclampsia, gestational diabetes, or bone fractures were noted in either of the two groups. CONCLUSION(S): In this pilot study, the use of LDA in combination with LMWH during pregnancy for the prevention of recurrent pregnancy loss in women with antiphospholipid syndrome seems to be as safe as UFH plus LDA. Large, randomized trials will be required to determine differences in outcome with LMWH and LDA compared with treatment with UFH combined with LDA in this group of patients. PMID- 15749500 TI - Catheter type does not affect the outcome of intrauterine insemination treatment: a prospective randomized study. AB - OBJECTIVE: To compare the main outcome of IUI with the Gynetics catheter (Gynetics Medical Products, Hamont-Achel, Belgium) or the Makler cannula (Sefi Medical Instruments, Haifa, Israel). DESIGN: Prospective, randomized study. SETTING: Infertility and endocrinology unit in a university hospital. PATIENT(S): Two hundred fifty-one infertile couples undergoing 784 consecutive IUI treatments. INTERVENTION(S): Patients were randomly assigned to undergo IUI treatment with either the Gynetics catheter (124 patients) or the Makler cannula (127 patients). MAIN OUTCOME MEASURE(S): Primary outcome measures were pregnancy and cumulative pregnancy rates. Secondary outcome measures were the ease of introduction of the catheter, the presence of bleeding, and semen regurgitation after removal of the catheter. RESULT(S): The use of the Gynetics catheter resulted in similar pregnancy (10.4% vs. 9.7%) and cumulative pregnancy rates (27.9% vs. 26.4%) as compared with the Makler cannula. Difficult introduction of the catheter was more frequent in the Gynetics group than in the Makler group (19.4% vs. 8.0%, respectively), but the frequency of bleeding did not differ between groups. Sample regurgitation was observed more often in the Makler group than in the Gynetics group (49.9% vs. 17.9%, respectively). CONCLUSION(S): A lower frequency of sperm regurgitation and a higher frequency of difficult introduction was observed when the Gynetics catheter was used. However, there was no significant difference in pregnancy rates with use of either the Gynetics catheter or the Makler cannula. PMID- 15749499 TI - Antiphospholipid antibodies prevent extravillous trophoblast differentiation. AB - OBJECTIVE: We investigated the hypothesis that antiphospholipid antibodies (aPL) have a detrimental effect on human extravillous trophoblast (EVT) differentiation into giant multinucleated cells "in vitro." DESIGN: The EVT were isolated from the placental chorion using enzymatic digestion and Percoll gradient centrifugation. After 24, 36, and 48 hours in culture, giant multinuclear cells (GMC) were identified by immunohistochemistry using antibodies to cytokeratin 7 and counted. SETTING: An academic research laboratory. PATIENT(S): Placentas were donated by women having an elective cesarean section for a normal pregnancy at term. MAIN OUTCOME MEASURE(S): This model was then used to investigate the effect of two different monoclonal aPL to beta2-glycoprotein 1 (IIC5 and ID2), and control mouse IgG antibody on EVT differentiation. RESULT(S): Freshly isolated EVT were nonproliferative but moved together losing their intervening cell walls and differentiated into GMC. Maximal numbers of GMC were detected after 48 hours of culture. The aPL, IIC5, and ID2 significantly inhibited GMC formation, whereas the mouse IgG control had no effect. CONCLUSION(S): Antiphospholipid antibodies can inhibit EVT differentiation and GMC formation "in vitro" suggesting that a failure of trophoblast differentiation and subsequent uteroplacental development may be an underlying pathology in antiphospholipid syndrome-associated pregnancy loss. PMID- 15749501 TI - Hysteroscopic polypectomy in 240 premenopausal and postmenopausal women. AB - OBJECTIVE: To ascertain the therapeutic efficacy and safety of hysteroscopic polypectomy in 240 premenopausal and postmenopausal patients. DESIGN: Retrospective study. SETTING: Tertiary university hospital. PATIENT(S): Two hundred forty patients with intrauterine endometrial polyps, who mostly suffered from abnormal uterine bleeding and infertility. INTERVENTION(S): Hysteroscopic polypectomy using various instruments including microscissors, grasping forceps, or electrosurgery either with a monopolar probe or a resectoscope. MAIN OUTCOME MEASURE(S): Operating time, amount of glycine absorption, complications, resumption of normal menstruation, cumulative pregnancy rate, and recurrent rate of polyps after hysteroscopic surgery. RESULT(S): Resectoscopic polypectomy needed more operating time, had more glycine absorption and complications, but less recurrence than other hysteroscopic techniques. The resectoscope had a 0% recurrence rate and that grasping forceps had a 15% recurrence rate. A total of 21 (8.7%) complications occurred, but no major complications were noted. After long-term follow-up of 9 years and 2 months, those with abnormal uterine bleeding resumed normal menstruation in 93.1% and those with infertility had a cumulative pregnancy rate of 42.3%. There was no statistical difference in reproductive outcome between patients having polyps < or = 2.5 cm and >2.5 cm. CONCLUSION(S): We found hysteroscopic polypectomy to be effective, safe, minimally invasive procedure with low rate and mild complications. Restoration of reproductive ability did not depend on the size of the removed lesion. Resectoscopic surgery is more preferable to prevent recurrence of polyps. PMID- 15749502 TI - Embryo afterloading: a refinement in embryo transfer technique that may increase clinical pregnancy. AB - OBJECTIVE: Given the importance of ET technique during assisted reproductive technology cycles, we evaluated the effect of embryo afterloading subsequent to placement of the ET catheter on pregnancy rates vs. a standard direct ET. DESIGN: Retrospective cohort analysis. SETTING: University-based assisted reproductive technology program. PATIENT(S): Patients undergoing a fresh nondonor day 3 ET by a single provider over a 1-year period. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical pregnancy. RESULT(S): One hundred twenty-seven patients met inclusion criteria, and the overall pregnancy rate was 46.5%. There was no difference between the two groups with respect to age, basal FSH, or number of embryos transferred. The ET method used was at the discretion of the provider. There was no difference between the two groups in the presence of blood on the transfer catheter. However, there were significantly more transfer catheters with mucus contamination in the direct transfer group (25.58% vs. 5.95%). The clinical pregnancy rate in the group with ET using the afterloading technique was higher than in the direct ET group (52.4% vs. 34.9%). CONCLUSION(S): There was a trend toward an increase in pregnancy rate when an embryo afterloading technique was used. A prospective randomized trial is needed to examine this issue. PMID- 15749503 TI - Thermachoice endometrial ablation in the outpatient setting, without local anesthesia or intravenous sedation: a prospective cohort study. AB - OBJECTIVE: To determine whether Thermachoice endometrial ablation (EA) is a safe and acceptable procedure when performed in the outpatient (OP) setting without local anesthesia or IV sedation. DESIGN: Prospective cohort (double group) study. SETTING: Hysteroscopy clinic of large UK hospital. PATIENT(S): Twenty-seven women with menorrhagia. INTERVENTION(S): The first cohort of women underwent Thermachoice EA without elective analgesia. The second cohort underwent Thermachoice after taking ibuprofen. MAIN OUTCOME MEASURE(S): To map pain scores and the requirement of "rescue analgesia." To assess speed of recovery and time away from home. RESULT(S): Thermachoice EA was successfully performed in the OP setting on 89% (n = 24) of women. Four women in the first cohort required rescue analgesia compared to none in the second. During the procedure there was little difference between the groups visual analogue style pain scores-with the overall score being 3.6 (range = 0-10). Postoperatively there was a lower mean pain score in the second cohort, with fewer women experiencing nausea and vomiting. Mean time away from home was 2.6 hours and mean time to make a full recovery was 3.3 days. Women required analgesia for 2.6 days, on average, and returned to their normal activities at this time. CONCLUSION(S): Thermachoice EA without local analgesia or IV sedation can be safely and successfully performed in the OP setting and if ibuprofen is taken preoperatively few women require "rescue analgesia." PMID- 15749504 TI - Effectiveness of two-step (consecutive) embryo transfer in patients who have two embryos on day 2: comparison with cleavage-stage embryo transfer. AB - OBJECTIVE: To evaluate the effectiveness, for patients who have only two embryos on day 2, of a two-step (consecutive) embryo transfer (ET) procedure in which a cleaved embryo is transferred on day 2 and a single blastocyst is transferred on day 5. DESIGN: Observational comparative study. SETTING: Private IVF clinic. PATIENT(S): Ninety two-step ET cycles were performed in patients who had two embryos on day 2 (two-step group). Ninety day-2 ET cycles were performed in age- and infertility-matched patients who had two embryos on day 2 (control group). INTERVENTION(S): Cleaved-ET, extended culture of one embryo, and a second transfer of a blastocyst. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): The pregnancy and implantation rates in the two-step group (respectively 33.3% and 17.2%) were significantly higher than those in the control group (18.9% and 9.4%). Thirty-nine of the patients in the two-step group (43.3%) could not proceed to the second step of ET because no viable blastocyst could be obtained, but four of them conceived anyway. CONCLUSION(S): Taking advantage of both day-2 ET and blastocyst transfer, two-step ET may be an effective option for ET in patients who have an insufficient number of embryos. PMID- 15749505 TI - Bipolar radio frequency endometrial ablation compared with balloon endometrial ablation in dysfunctional uterine bleeding: impact on patients' health-related quality of life. AB - OBJECTIVE: To compare health-related quality of life (HRQoL) after bipolar radio frequency ablation and thermal balloon ablation in women with dysfunctional uterine bleeding. DESIGN: Randomized clinical trial. SETTING: Teaching hospital. PATIENT(S): Women suffering from dysfunctional uterine bleeding. INTERVENTION(S): Bipolar radio frequency ablation and thermal balloon ablation. MAIN OUTCOME MEASURE(S): Patients were asked to complete HRQoL questionnaires at baseline, and at 2 days, 2 weeks, 3 months, 6 months, and 12 months after surgery. The questionnaires contained the medical outcomes study Short-Form 36 (SF-36), the Self-rating Depression Scale, the Rotterdam Symptom Checklist, State-Trait Anxiety Inventory, and a structured clinical history questionnaire. RESULT(S): Data on HRQoL were available on at least two different time points in 115 of 126 randomized patients. HRQoL improved significantly over time in both groups, except for the domain of general health in the SF-36. None of the dimensions showed a significant difference between both groups, neither was there a significant interaction between time and treatment effect. CONCLUSION(S): Both methods significantly improved HRQoL in women with dysfunctional uterine bleeding. However, despite better amenorrhea and satisfaction rates after bipolar radio frequency ablation, there was no difference in HRQoL between the two groups. PMID- 15749506 TI - Laparoscopic treatment of pelvic pain associated with minimal and mild endometriosis with use of the Helica Thermal Coagulator. AB - OBJECTIVE: To investigate the clinical efficacy and safety of Helica Thermal Coagulator (TC) in the treatment of pelvic pain associated with minimal (stage I) and mild (stage II) endometriosis. DESIGN: A clinical observational study. SETTING: A referral center for laparoscopic treatment of endometriosis. PATIENT(S): Eighty-one women with pelvic pain symptoms associated with minimal and mild endometriosis diagnosed at laparoscopy. INTERVENTION(S): Helica TC to treat endometriotic lesions. The revised American Fertility Society (rAFS) classification was used to stage endometriosis. Pain symptoms and patient satisfaction were assessed subjectively at 3 and 6 months follow-up. MAIN OUTCOME MEASURE(S): Improvement or relief of pelvic pain symptoms, and intra- or postoperative complications. RESULT(S): A total of 79 women completed the study to 6 months follow-up. At 3 months, 59 (74.7%) women reported resolution and satisfactory improvement of symptoms, whereas 20 (25.3%) women continued to experience painful symptoms. At 6 months, 69 (87.4%) women reported resolution and satisfactory improvement of symptoms, whereas 9 (11.4%) women reported no changes and 1 (1.2%) woman experienced worsening symptoms. No significant differences were found between minimal and mild disease. No side effects or surgical complications occurred. CONCLUSION(S): Meaningful improvements and relief in clinical symptoms can be obtained with conservative laparoscopic surgery. Helica TC is a simple, effective, and safe device for the treatment of pelvic pain in women with stages I and II endometriosis. This approach requires further evaluation as part of randomized controlled trials. PMID- 15749507 TI - Ectopic mesothelial cell proliferation in cervical lymph nodes after severe ovarian hyperstimulation syndrome. AB - OBJECTIVE: To report a case of ectopic mesothelial proliferation in cervical lymph nodes after severe ovarian hyperstimulation syndrome. DESIGN: Case report. SETTING: University-affiliated teaching hospital. PATIENT(S): A 42-year-old woman underwent a successful IVF attempt and developed severe ovarian hyperstimulation syndrome and pathologically enlarged cervical lymph nodes. INTERVENTION(S): Paracentesis, cervical lymph node biopsy followed by cytology, histology, and immunohistochemistry. MAIN OUTCOME MEASURE(S): Resolution of symptoms, pregnancy outcome, correct pathological diagnosis. RESULT(S): Paracentesis resulted in resolution of symptoms of ovarian hyperstimulation. The diagnosis of ectopic mesothelial proliferation in cervical lymph nodes was made after immunohistochemical examination of cervical lymph nodes. The pregnancy progressed normally, and at 40 weeks the patient spontaneously delivered a healthy son weighing 3,060 g. CONCLUSION(S): This case describes ectopic mesothelial cell proliferation localized in and around cervical lymph nodes occurring at 9 weeks' gestation in a patient who earlier developed severe ovarian hyperstimulation syndrome 10 days after ET. Ectopic mesothelial cell proliferation in lymph nodes is an extremely rare event, and this is the first case described after ovarian hyperstimulation. Familiarity with this event is important for the clinician as well as for the pathologist in preventing the misdiagnosis of malignancy. PMID- 15749508 TI - Monochorionic triplet gestation after in vitro fertilization using donor oocytes: case report and review. AB - OBJECTIVE: To report on a patient with a monochorionic triamnionic triplet pregnancy after IVF with donor oocytes. DESIGN: Case report. SETTING: Academic tertiary care hospital. PATIENT(S): A 42-year-old woman who underwent IVF with donor oocytes. INTERVENTION(S): After failed IVF attempts, the patient chose to undergo treatment with donor oocytes. Her 23-year-old oocyte donor underwent standard controlled ovarian hyperstimulation. Retrieved oocytes were fertilized in vitro, and two embryos were transferred at the blastocyst stage. MAIN OUTCOME MEASURE(S): Intrauterine pregnancy with single gestational sac and three fetal poles with cardiac activity. RESULT(S): After extensive counseling with perinatologists about pregnancy complications, the patient elected to terminate at 10 weeks of gestation. CONCLUSION(S): Several processes have been suggested to explain the increase in monozygotic twinning after IVF. These factors include advanced maternal age, superovulation, manipulation of the zona pellucida, and prolonged culture. It is possible that other factors may also play a role, especially in high-order monozygotic multiple pregnancies. All patients should be informed of the potential risk of a high-order multiple pregnancy after IVF, even when only one or two embryos are transferred. PMID- 15749509 TI - Obstetrical complications in gestational carrier pregnancies. AB - OBJECTIVE: To report two cases of severe obstetrical complications in gestational carrier pregnancies and to review our clinical experience and compare our results with those reported in the literature. DESIGN: Retrospective analysis. SETTING: A university IVF program. PATIENT(S): Women without a functioning uterus or those whose pregnancy would exacerbate a medical condition were enrolled in the gestational carrier pregnancy program. INTERVENTION(S): IVF cycles using oocytes from genetic mothers (or oocyte donors) were performed, with ET to gestational carriers. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates, obstetrical complications, and neonatal outcomes. RESULT(S): Ten couples underwent a total of 13 cycles using gestational carriers. A clinical pregnancy rate of 69% (9/13) was achieved. An intrapartum hysterectomy and a late puerperal hysterectomy were required because of severe obstetrical complications. The late puerperal hysterectomy was performed for placenta accreta in a triplet gestation. This carrier sustained multiple cerebral infarcts and blindness. One triplet infant died secondary to a hypoplastic left ventricle and complications of prematurity. A second gestational carrier with a singleton gestation underwent a hysterectomy for a uterine rupture, and the infant has cerebral palsy. CONCLUSION(S): The past medical and obstetrical histories of potential gestational carriers must be closely scrutinized, and candidates must be thoroughly counseled about the potential risks involved in the procedure. PMID- 15749510 TI - Uterine devascularization and subsequent major intrauterine synechiae and ovarian failure. AB - OBJECTIVE: To report a case of major intrauterine synechiae associated with ovarian function failure secondary to uterine devascularization. DESIGN: Case report. SETTING: Departments of gynecology and obstetrics in two hospitals in France. PATIENT(S): A 29-year-old woman, para 1, presenting severe postpartum hemorrhage. INTERVENTION(S): Bilateral uterine, round ligament, utero-ovarian, and ovarian arteries ligation. MAIN OUTCOME MEASURE(S): Clinical and hormonal status, computed tomography scan, hysterosalpingography, laparoscopy, hysteroscopy, and histological examination. RESULT(S): Twelve months after uterine devascularization, the patient presented major intrauterine synechiae, absence of endometrium, and ovarian function failure. CONCLUSION(S): Uterine devascularization may be responsible for infertility as a result of intrauterine synechiae and ovarian function failure. PMID- 15749511 TI - Endometriosis in premenarcheal girls who do not have an associated obstructive anomaly. AB - OBJECTIVE: To identify endometriosis as a cause of chronic pelvic pain in premenarcheal girls without an obstructive anomaly of the reproductive tract. DESIGN: A case series. SETTING: Tertiary-care academic pediatric hospital. PATIENT(S): Five premenarcheal girls with chronic (>6 months) pelvic pain and negative gastrointestinal workup. INTERVENTION(S): Laparoscopic biopsies and resection, laser and/or cautery of all visible lesions. MAIN OUTCOME MEASURE(S): Visual presence of endometriosis and decrease in pelvic pain after destruction of lesions. RESULT(S): All subjects had laparoscopy with the identification of clear, red, and/or white lesions consistent with endometriosis based on the standard American Society for Reproductive Medicine Classification of Endometriosis. Postoperatively, all of the girls had marked improvement of their pelvic pain based on self-reported pain scales. Two of the subjects had subsequent repeat laparoscopies 6 and 8 years after their initial surgery, which revealed pathology-proven endometriosis. CONCLUSION(S): Endometriosis should be part of the differential diagnosis of chronic pelvic/lower abdominal pain in premenarcheal girls. The presence of lesions visibly consistent with endometriosis in premenarcheal girls without an obstructive anomaly supports the concept that some cases of endometriosis may result from an etiology other than retrograde menses. PMID- 15749512 TI - Two successive pregnancies after ursodeoxycholic acid therapy in a previously infertile woman with antimitochondrial antibody-negative primary biliary cirrhosis. AB - OBJECTIVE: To describe the benefit of ursodeoxycholic acid (UDCA) for the initiation and completion of a successful pregnancy in a previously infertile woman with primary biliary cirrhosis. DESIGN: Case report. SETTING: A university hospital with relevant departments. PATIENT(S): A 29-year-old woman with primary biliary cirrhosis and failure to conceive for 6 years. INTERVENTION(S): Establishment of diagnosis with a liver biopsy, pretreatment of patient with UDCA before conception, and continuation of UDCA after first trimester until term. UDCA was used in the second pregnancy again after the first trimester. MAIN OUTCOME MEASURE(S): Achievement of a safe conception and full-term pregnancy. RESULT(S): Two consecutive successful pregnancies, a healthy 3,250-g male infant and a healthy 3,000-g female infant. The second conception occurred in a period without the use of UDCA, implicating a latent beneficial effect of either UDCA or the previous pregnancy via some possible immune mechanism. CONCLUSION(S): Ursodeoxycholic acid could help achieve conception in infertile women with primary biliary cirrhosis. The use of UDCA after the first trimester is shown to be safe in two consecutive pregnancies. Although it cannot be conclusive, the unintentional use of UDCA in the first 20 days after conception did not result in any teratogenicity in the first child. PMID- 15749513 TI - Isochromosome of Yp in a man with Sertoli-cell-only syndrome. AB - OBJECTIVE: To address phenotype/genotype correlation in a man with i(Y)(p10). DESIGN: Case report. SETTING: University-based reproductive genetics laboratory. PATIENT(S): A 27-year-old azoospermic man with i(Y)(p10), relatively normal stature, and testicular Sertoli-cell-only syndrome. INTERVENTION(S): Testicular biopsy, cytogenetic study, Y-chromosome deletion mapping analysis, fluorescence in situ hybridization (FISH). MAIN OUTCOME MEASURE(S): Expression of Y-chromosome genes. RESULT(S): We have identified one azoospermic man with i(Y)(p10) of 312 Taiwanese men presenting with a severe spermatogenic defect. Y-chromosome deletion mapping analysis confirmed deletions of all Yq sequences, including a putative growth controlling gene. Fluorescence in situ hybridization (FISH) analysis showed duplication of Yp material. The patient had normal stature considering midparental height. He also had no germ cells in the testicular tissue (Sertoli-cell-only syndrome) resulting from the loss of azoospermia factor in Yq. CONCLUSION(S): Among structural rearrangements of the Y-chromosome, the isochromosome of Yp occurs very rarely. This case is the first reported case of an isochromosome Yp with a detailed description of testicular histology and body height. PMID- 15749514 TI - Spontaneous pregnancy after bilateral salpingectomy. AB - OBJECTIVE: To report a rare case of spontaneous pregnancy following bilateral salpingectomy. DESIGN: Case report. SETTING: Maternity unit at a district general hospital in the United Kingdom. PATIENT(S): A healthy 38-year-old woman with a history of bilateral salpingectomy. INTERVENTION(S): Ultrasound scan and serum beta hcG used to diagnose the pregnancy and for follow-up. MAIN OUTCOME MEASURE(S): Recognition and successful management of the pregnancy. RESULT(S): The patient was diagnosed with a noncontinuing pregnancy that resolved spontaneously. CONCLUSION(S): A MEDLINE search revealed that this is the first reported case of spontaneous pregnancy following bilateral salpingectomy in the English-language literature and possibly only the second in world literature. PMID- 15749516 TI - Impact of isolated teratozoospermia on the outcome of intrauterine insemination. AB - One thousand six hundred forty-one IUI cycles performed in 615 couples were categorized, depending on the semen analysis of the male partner, in three groups of: normozoospermia, teratozoospermia, and male factor infertility. Clinical pregnancies and live births per cycle were significantly decreased in the teratozoospermia group when compared to the normozoospermia group, with the exception of the first IUI attempt (comparable outcomes), whereas the cumulative live birth rate after four IUI attempts was significantly lower in the teratozoospermia and male factor infertility groups. PMID- 15749515 TI - Spontaneous pregnancy and birth of a normal female from a woman with Turner syndrome and elevated gonadotropins. AB - OBJECTIVE: To provide data for pregnancies in girls with Turner syndrome. Only 5% 10% of TS girls undergo spontaneous puberty and have menses. Spontaneous pregnancy occurs in 2%-7% of TS girls and is associated with a high rate of miscarriages, stillbirths, malformations, and chromosomal aberrations. Besides fetal problems, pregnancy in TS girls is of high risk for the mothers as well. DESIGN: Case report. SETTING: Academic unit. PATIENT(S): One patient, now aged 28 years, was referred for short stature at age 13 years after an operation for coarctation of the aorta. The karyotype was 88% 45X, 5% 46XX, 5% 47XXX, 2% XO+Fr. The gonadotropin values at presentation and on follow-up ranged from high normal to high levels. Pubertal development and menses occurred spontaneously. At the age of 20 and 21 years, dominant follicles of 14 and 17 mm, respectively, were found on sonography. She conceived spontaneously at the age of 26 years and had an uneventful pregnancy, giving birth to a normal girl with normal chromosome constitution and birth weight of 2,800 g. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy outcome. RESULT(S): Successful spontaneous pregnancy. CONCLUSION(S): Bearing in mind the serious problems of fertility and pregnancy outcome encountered in TS girls, we considered such a rare escape from the expected course of biological events to be worth reporting. PMID- 15749517 TI - Q188R, K285N, and N314D mutation-associated alleles in the galactose-1-phosphate uridyltransferase gene and female infertility. AB - In a retrospective case-control study, the frequencies of Q188R, K285N, N314D, and IVS5-24G>A mutations were determined with the use of polymerase chain reaction and restriction fragment length polymorphism in the group of infertile women and the controls. No statistically significant differences were observed in the allele frequencies between the infertile women and control groups. PMID- 15749518 TI - How do infertile couples choose their IVF centers? An Internet-based survey. AB - We evaluated and compared different factors affecting a patient's choice of initial and subsequent in vitro fertilization (IVF) clinics using an Internet based survey. The two highest-ranking factors affecting both initial and subsequent clinic choices were the success rate of the clinic and the quality of the service provided. PMID- 15749519 TI - Algorithm to predict assisted reproductive technology pregnancy outcome reveals minimal embryo synergy. AB - A mathematical model was developed to calculate the implantation probability for individual embryos based on the pregnancy outcome of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cases with multiple embryos transferred. This model was used to calculate implantation probabilities of embryos of 31 morphological types using the outcome of 1,200 IVF/ICSI cases. The algorithm was validated by comparing the calculated pregnancy probability and multiple pregnancy probability with the actual outcome of 281 separate IVF/ICSI cases. Finally, an estimation of embryo synergy was calculated. PMID- 15749520 TI - Effect of halving the daily dose of triptorelin at the start of ovarian stimulation on hormone serum levels and the outcome of in vitro fertilization. AB - Halving the standard daily dose of triptorelin at the start of ovarian stimulation in down-regulated women stimulated with recombinant FSH is enough for pituitary suppression and was associated with higher LH serum concentrations in the follicular phase. However, this did not translate into higher serum concentrations of androstenedione and E2 and had no significant effect on ovarian response and the outcome of IVF/intracytoplasmic sperm injection. PMID- 15749521 TI - An indirect quantitative method for the assessment of angiogenesis in human tissues. AB - We set up an ELISA that measures the concentration of von Willebrand factor (vWF) in human endometrial tissue and found a significant correlation with the mean vessel counts in vWF- and CD31-immunostained tissue sections. This ELISA allows an objective and quantitative evaluation of the vascular state in the endometrium and could be used as a method to evaluate the angiogenic state in other tissues. PMID- 15749523 TI - T homozygote and allele of epidermal growth factor receptor 2073 gene polymorphism are associated with higher susceptibility to endometriosis and leiomyomas. AB - Epidermal growth factor receptor (EGFR) is a regulator of angiogenesis and mediator of sex steroid-induced cell growth and differentiation. We observed that EGFR gene 2073*T-related genotypes and allele are associated with higher susceptibilities to endometriosis and leiomyoma. PMID- 15749522 TI - Angiostatic agents prevent the development of endometriosis-like lesions in the chicken chorioallantoic membrane. AB - A prospective study was performed to determine the effects of the angiostatic compounds anti-hVEGF antibody, TNP-470, endostatin, and anginex on the vascularization and on endometriosis-like lesion formation in the chicken chorioallantoic membrane model. Endometriosis-like lesion formation was significantly impaired after treatment with angiostatic agents, which was associated with decreased vessel densities in the surrounding chorioallantoic membrane and more necrosis in the endometriosis-like lesions. PMID- 15749524 TI - Characterization of oxidative stress status by evaluation of reactive oxygen species levels in whole semen and isolated spermatozoa. AB - We defined the basal levels of reactive oxygen species (ROS) in normal donors in neat (whole unprocessed) semen specimens, and in mature and immature spermatozoa isolated by a double-density gradient technique. In addition, we demonstrated that the ROS levels were significantly lower in neat semen compared with washed spermatozoa. The reference values of ROS in neat semen and mature spermatozoa can be used to define the pathologic levels of ROS in infertile men and may guide in therapeutic interventions. PMID- 15749525 TI - A Markov model of the cost-effectiveness of human-derived follicle-stimulating hormone (FSH) versus recombinant FSH using comparative clinical trial data. AB - This study compared the cost and effectiveness of highly purified, human-derived follicle-stimulating hormone (FSH) (Bravelle) to recombinant FSH (Follistim) using Markov modeling and Monte Carlo simulation. One IVF treatment cycle resulted in costs of 11,584 dollars +/- 211 dollars for human-derived FSH and 12,762 dollars +/- 170 dollars for recombinant FSH, while three treatment cycles, holding the transition probabilities of the first cycle constant for the next two cycles, resulted in costs of 22,712 dollars +/- 1,107 dollars for human-derived FSH and 24,935 dollars +/- 1,205 dollars for recombinant FSH. PMID- 15749526 TI - Influence of reactive oxygen species produced by activated leukocytes at the level of apoptosis in mature human spermatozoa. AB - Human spermatozoa incubated with polymorphonuclear leukocytes and Escherichia coli showed a significant increase in the annexin V binding (P<.05). This increase was similar to that obtained when spermatozoa were incubated with polymorphonuclear leukocytes activated by phorbol-12-myristate-13-acetate. PMID- 15749527 TI - The antral follicle count is a better marker than basal follicle-stimulating hormone for the selection of older patients with acceptable pregnancy prospects after in vitro fertilization. AB - This observational study shows that the antral follicle count is a better predictor of ongoing pregnancy in IVF patients aged >38 years of age than is basal FSH. Patients aged <44 years with a normal antral follicle count still have acceptable pregnancy rates after IVF and therefore deserve treatment. PMID- 15749529 TI - Endometriosis at 2nd surgery--residual or recurrent disease. PMID- 15749530 TI - Quality of life after excision of endometriosis. PMID- 15749532 TI - The National Atmospheric Deposition Program: 25 years of monitoring in support of science and policy: an ammonia workshop: the state of science and future needs. PMID- 15749534 TI - Trends in atmospheric ammonium concentrations in relation to atmospheric sulfate and local agriculture. AB - Ammonium (NH(4)(+)) concentrations in air and precipitation at the Institute of Ecosystem Studies (IES) in southeastern New York, USA declined over an 11-year period from 1988 to 1999, but increased from 1999 to 2001. These trends in particulate NH(4)(+) correlated well with trends in particulate SO(4)(2-) over the 1988-2001 period. The NH(4)(+) trends were not as well correlated with local cattle and milk production, which declined continuously throughout the period. This suggests that regional transport of SO(4)(2-) may have a greater impact on concentrations of NH(4)(+) and subsequent deposition than local agricultural emissions of NH(3). Ammonium concentrations in precipitation correlated significantly with precipitation SO(4)(2-) concentrations for the 1984-2001 period although NH(4)(+) in precipitation increased after 1999 and SO(4)(2-) in precipitation continued to decline after 1999. The correlation between NH(4)(+) and SO(4)(2-) was stronger for particulates than for precipitation. Particulate NH(4)(+) concentrations were also correlated with particulate SO(4)(2-) concentrations at 31 of 35 eastern U.S. CASTNet sites that had at least 10 years of data. Air concentrations of NH(4)(+) and SO(4)(2-) were more strongly correlated at the sites that were located within an agricultural landscape than in forested sites. At most of the sites there was either no trend or a decrease in NH(4)(+) dry deposition during the 1988-2001 period. The sites that showed an increasing trend in NH(4)(+) dry deposition were generally located in the southeastern U.S. The results of this study suggest that, in the northeastern U.S., air concentrations of NH(4)(+) and subsequent deposition may be more closely linked to SO(4)(2-) and thus SO(2) emissions than with NH(3) emissions. These results also suggest that reductions in S emissions have reduced NH(4)(+) transport to and NH(4)(+)-N deposition in the Northeast. PMID- 15749533 TI - Spatial and temporal trends of precipitation chemistry in the United States, 1985 2002. AB - A Seasonal Kendall Trend (SKT) test was applied to precipitation-weighted concentration data from 164 National Atmospheric Deposition Program National Trends Network (NADP/NTN) sites operational from 1985 to 2002. Analyses were performed on concentrations of ammonium, sulfate, nitrate, dissolved inorganic nitrogen (DIN, sum of nitrogen from nitrate and ammonium), and earth crustal cations (ECC, sum of calcium, magnesium, and potassium). Over the 18-year period, statistically significant (p< or =0.10) increases in ammonium concentrations occurred at 93 sites (58%), while just three sites had statistically significant ammonium decreases. Central and northern Midwestern states had the largest ammonium increases. The generally higher ammonium concentrations were accompanied by significant sulfate decreases (139 sites, 85%), and only one significant increase which occurred in Texas. In the west central United States there were significant nitrate increases (45 sites, 27%), while in the northeastern United States there were significant decreases (25 sites, 15%). Significant DIN decreases were observed in the northeastern United States (11 sites, 7%); elsewhere there were significant increases at 75 sites (46%). ECC decreased significantly at 65 sites (40%), predominantly in the central and southern United States, and increased at 11 sites (7%). The concentrations of sulfate, nitrate, and ammonium in precipitation have changed markedly over the time period studied. Such trends indicate changes in the mix of gases and particles scavenged by precipitation, possibly reflecting changes in emissions, atmospheric chemical transformations, and weather patterns. PMID- 15749535 TI - Nonlinear regression and ARIMA models for precipitation chemistry in East Central Florida from 1978 to 1997. AB - Continuous monitoring of precipitation in East Central Florida has occurred since 1978 at a sampling site located on the University of Central Florida (UCF) campus. Monthly volume-weighted average (VWA) concentration for several major analytes that are present in precipitation samples was calculated from samples collected daily. Monthly VWA concentration and wet deposition of H(+), NH(4)(+), Ca(2+), Mg(2+), NO(3)(-), Cl(-) and SO(4)(2-) were evaluated by a nonlinear regression (NLR) model that considered 10-year data (from 1978 to 1987) and 20 year data (from 1978 to 1997). Little change in the NLR parameter estimates was indicated among the 10-year and 20-year evaluations except for general decreases in the predicted trends from the 10-year to the 20-year fits. Box-Jenkins autoregressive integrated moving average (ARIMA) models with linear trend were considered as an alternative to the NLR models for these data. The NLR and ARIMA model forecasts for 1998 were compared to the actual 1998 data. For monthly VWA concentration values, the two models gave similar results. For the wet deposition values, the ARIMA models performed considerably better. PMID- 15749536 TI - Overview and assessment of techniques to measure ammonia emissions from animal houses: the case of the Netherlands. AB - In order to comply with the ammonia (NH(3)) emission reduction assigned to the Netherlands development of new measures are needed, which should be supported by fast and accurate measurements to arrive at new estimates of the NH(3) emission from each agricultural source. This paper gives an overview of the current methods used in the Netherlands to measure NH(3) emissions from animal houses, and provides alternative methods for some particular situations. For mechanically ventilated animal houses, passive flux samplers placed in the ventilation shafts of the animal house are presented as alternative to measure a larger number of animal houses (replicates) with the same housing system at a low price. For naturally ventilated animal houses, when mixing in the animal house is not good enough to allow measurements within the animal house (internal tracer gas ratio method), two measurement methods are discussed: the Gaussian plume dispersion model, which is usually not suitable for agricultural situations, and the flux frame method, which is not always applicable because of distortion of the flow around the building. Finally, for animal houses with outside yards for the animals, there are at this moment no methods available to measure the NH(3) emissions from these complex situations, although quick box methods (for the outside yards) and a combination of a backward Lagrangian stochastic model with open-path concentration measurements with a tunable diode laser (TDL), look promising. PMID- 15749537 TI - Measuring ammonia emissions from land applied manure: an intercomparison of commonly used samplers and techniques. AB - A number of techniques have been developed to quantify ammonia (NH(3)) emissions following land application of manure or fertiliser. In this study, coefficients of variation were determined for three commonly used field techniques (mass balance integrated horizontal flux, wind tunnels and the equilibrium concentration technique) for measuring emissions from a range of manure types. Coefficients of variation (CV) for absorption flasks, passive flux samplers and passive diffusion samplers were 21, 10 and 14%, respectively. In comparative measurements, concentrations measured using passive flux samplers and absorption flasks did not differ significantly, but those measured using passive diffusion samplers were on average 1.8 times greater. The mass balance technique and wind tunnels gave broadly similar results in two out of four field tests. Overexposure of passive diffusion samplers for some sampling periods meant that estimation of cumulative NH(3) emission using the equilibrium concentration technique in the field tests could not be made. For cumulative NH(3) emissions, CVs were in the range of 23-52, 46-74 and 21-39% for the mass balance, wind tunnel and equilibrium concentration techniques, respectively. Lower CVs were associated with measurements following slurry compared with solid manure applications. Our conclusions from this study are that for the measurement of absolute emissions the mass balance technique is to be preferred, and for small-plot comparative measurements the wind tunnel system is preferred to the equilibrium concentration technique. PMID- 15749538 TI - Managing ammonia emissions from livestock production in Europe. AB - Around 75% of European ammonia (NH(3)) emissions come from livestock production. Emissions occur at all stages of manure management: from buildings housing livestock; during manure storage; following manure application to land; and from urine deposited by livestock on pastures during grazing. Ammoniacal nitrogen (total ammoniacal-nitrogen, TAN) in livestock excreta is the main source of NH(3). At each stage of manure management TAN may be lost, mainly as NH(3), and the remainder passed to the next stage. Hence, measures to reduce NH(3) emissions at the various stages of manure management are interdependent, and the accumulative reduction achieved by combinations of measures is not simply additive. This TAN-flow concept enables rapid and easy estimation of the consequences of NH(3) abatement at one stage of manure management (upstream) on NH(3) emissions at later stages (downstream), and gives unbiased assessment of the most cost-effective measures. We conclude that rapid incorporation of manures into arable land is one of the most cost-effective measures to reduce NH(3) emissions, while covering manure stores and applying slurry by band spreader or injection are more cost-effective than measures to reduce emissions from buildings. These measures are likely to rank highly in most European countries. PMID- 15749539 TI - Spatial and temporal variability of the overall error of National Atmospheric Deposition Program measurements determined by the USGS collocated-sampler program, water years 1989-2001. AB - Data from the U.S. Geological Survey (USGS) collocated-sampler program for the National Atmospheric Deposition Program/National Trends Network (NADP/NTN) are used to estimate the overall error of NADP/NTN measurements. Absolute errors are estimated by comparison of paired measurements from collocated instruments. Spatial and temporal differences in absolute error were identified and are consistent with longitudinal distributions of NADP/NTN measurements and spatial differences in precipitation characteristics. The magnitude of error for calcium, magnesium, ammonium, nitrate, and sulfate concentrations, specific conductance, and sample volume is of minor environmental significance to data users. Data collected after a 1994 sample-handling protocol change are prone to less absolute error than data collected prior to 1994. Absolute errors are smaller during non winter months than during winter months for selected constituents at sites where frozen precipitation is common. Minimum resolvable differences are estimated for different regions of the USA to aid spatial and temporal watershed analyses. PMID- 15749540 TI - Deposition monitoring networks: what monitoring is required to give reasonable estimates of ammonia/ammonium? AB - Deposition is one of the main loss terms for ammonia and ammonium from the atmosphere. It is also the input for ecosystems that can lead to drastic changes and effects. Deposition networks are needed to evaluate the need and the effect of policies to reduce nitrogen emissions, but also for studying deposition parameters and for developing deposition models. As with ambient concentrations of ammonia, deposition, especially dry deposition, varies strongly in space and in time. Furthermore, the bi-directional surface-atmosphere exchange of ammonia makes the combination of ambient concentration measurements with inferential models inadequate. Developing deposition monitoring networks with reasonable accuracy and representativeness is therefore not straightforward. In Europe several projects have addressed deposition monitoring. From these results it is concluded that a monitoring strategy should consist of a network with a limited amount of super sites combined with a larger number of sites where low cost methods are applied, together with models for generalisation. PMID- 15749541 TI - Atmospheric nitrogen inputs to the Delaware Inland Bays: the role of ammonia. AB - A previous assessment of nitrogen loading to the Delaware Inland Bays indicates that atmospheric deposition provides 15-25% of the total, annual N input to these estuaries. A large and increasing fraction of the atmospheric wet flux is NH(4)(+), which for most aquatic organisms represents the most readily assimilated form of this nutrient. Particularly noteworthy is a 60% increase in the precipitation NH(4)(+) concentration at Lewes, DE over the past 20 years, which parallels the increase in poultry production on the Delmarva Peninsula over this period (currently standing at nearly 585 million birds annually). To further examine the relationship between local NH(3) emissions and deposition, biweekly integrated gaseous NH(3) concentrations were determined using Ogawa passive samplers deployed at 13 sampling sites throughout the Inland Bays watershed over a one-year period. Annual mean concentrations at the 13 sites ranged from <0.5 microg NH(3)m(-3) to >6 microg NH(3)m(-3), with a mean of 1.6+/-1.0 microg NH(3)m(-3). At most sites, highest NH(3) concentrations were evident during spring and summer, when fertilizer application and poultry house ventilation rates are greatest, and seasonally elevated temperatures induce increased rates of microbial activity and volatilization from soils and animal wastes. The observed north-to-south concentration gradient across the watershed is consistent with the spatial distribution of poultry houses, as revealed by a GIS analysis of aerial photographs. Based on the average measured NH(3) concentration and published NH(3) deposition rates to water surfaces (5-8 mm s(-1)), the direct atmospheric deposition of gaseous NH(3) to the Inland Bays is 3.0-4.8 kg ha( 1)yr(-1). This input, not accounted for in previous assessments of atmospheric loading to the Inland Bays, would effectively double the estimated direct dry deposition rate, and is on par with the NO(3)(-) and NH(4)(+) wet fluxes. A second component of this study examined spatial differences in NO(3)(-) and NH(4)(+) wet deposition within the Inland Bays watershed. In a pilot study, precipitation composition at the Lewes NADP-AIRMoN site (DE 02) was compared with that at a satellite site established at Riverdale on the Indian River Estuary, approximately 21 km southwest. While the volume-weighted mean precipitation NO(3)(-) concentrations did not differ significantly between sites, the NH(4)(+) concentration observed at Riverdale (26.3 micromoles L(-1)) was 73% greater than at Lewes (15.2 micromoles L(-1)). More recently, a NADP site was established at Trap Pond, DE (DE 99), which was intentionally located within the region of intense poultry production. A comparison of the initial two years (6/2001-5/2003) of precipitation chemistry data from Trap Pond with other nearby NADP-AIRMoN sites (Lewes and Smith Island) reveals fairly homogeneous NO(3)(-) wet deposition, but significant spatial differences ( approximately 60%) in the NH(4)(+) wet flux. Overall, these results suggest that local emissions and below cloud scavenging provide a significant contribution to regional atmospheric N deposition. PMID- 15749542 TI - Improved daily precipitation nitrate and ammonium concentration models for the Chesapeake Bay Watershed. AB - Daily precipitation nitrate and ammonium concentration models were developed for the Chesapeake Bay Watershed (USA) using a linear least-squares regression approach and precipitation chemistry data from 29 National Atmospheric Deposition Program/National Trends Network (NADP/NTN) sites. Only weekly samples that comprised a single precipitation event were used in model development. The most significant variables in both ammonium and nitrate models included: precipitation volume, the number of days since the last event, a measure of seasonality, latitude, and the proportion of land within 8km covered by forest or devoted to industry and transportation. Additional variables included in the nitrate model were the proportion of land within 0.8km covered by water and/or forest. Local and regional ammonia and nitrogen oxide emissions were not as well correlated as land cover. Modeled concentrations compared very well with event chemistry data collected at six NADP/AirMoN sites within the Chesapeake Bay Watershed. Wet deposition estimates were also consistent with observed deposition at selected sites. Accurately describing the spatial distribution of precipitation volume throughout the watershed is important in providing critical estimates of wet-fall deposition of ammonium and nitrate. PMID- 15749543 TI - Chemometrics methods for the investigation of methylmercury and total mercury contamination in mollusks samples collected from coastal sites along the Chinese Bohai Sea. AB - The development and application of chemometrics methods, principal component analysis (PCA), cluster analysis and correlation analysis for the determination of methylmercury (MeHg) and total mecury (HgT) in gastropod and bivalve species collected from eight coastal sites along the Chinese Bohai Sea are described. HgT is directly determined by atomic fluorescence spectrometry (AFS), while MeHg is measured by a laboratory established high performance liquid chromatography atomic fluorescence spectrometry system (HPLC-AFS). One-way ANOVA and cluster analysis indicated that the bioaccumulation of Rap to accumulate Hg was significantly (P<0.05) different from other mollusks. Correlation analysis shows that there is linear relationship between MeHg and HgT in mollusks samples collected from coastal sites along the Chinese Bohai Sea, while in mollusks samples collected from Hongqiao market in Beijing City, there is not any linear relationship. PMID- 15749544 TI - Effects of increased deposition of atmospheric nitrogen on an upland Calluna moor: N and P transformations. AB - This study determined the effects of increased N deposition on rates of N and P transformations in an upland moor. The litter layer and the surface of the organic Oh horizon were taken from plots that had received long-term additions of ammonium nitrate at rates of 40, 80 and 120 kg N ha(-1) yr(-1). Net mineralisation processes were measured in both field and laboratory incubations. Soil phosphomonoesterase (PME) activity and rates of N(2)O release were measured in laboratory incubations and root-surface PME activity measured in laboratory microcosms using Calluna vulgaris bioassay seedlings. Net mineralisation rates were relatively slow, with net ammonification consistently stimulated by N addition. Net nitrification was marginally stimulated by N addition in the laboratory incubation. N additions also increased soil and root-surface (PME) activity and rates of N(2)O release. Linear correlations were found between litter C:N ratio and all the above processes except net nitrification in field incubations. When compared with data from a survey of European forest sites, values of litter C:N ratio were greater than a threshold below which substantial, N input-related increases in net nitrification rates occurred. The maintenance of high C:N ratios with negligible rates of net nitrification was associated with the common presence of ericaceous litter and a mor humus layer in both this moorland as well as the forest sites. PMID- 15749545 TI - Risk to breeding success of waterbirds by contaminants in Hong Kong: evidence from trace elements in eggs. AB - Concentrations of 17 trace elements (As, Se, Rb, Sr, Ag, Sb, Cs, Tl, Pb, V, Cr, Mn, Co, Cu, Zn, Cd, and Hg) were determined in eggs of little egrets (Egretta garzetta), black-crowned night herons (Nycticorax nycticorax) and bridled terns (Sterna anaethetus) from northwestern and northeastern coastal areas of Hong Kong. Results indicated significant differences among the three species. Eggs of little egrets and black-crowned night herons appeared to be good bioindicators for monitoring selected elements (As, Pb, Zn Cr, Cu, and V) due to consistent correlations between metal concentrations in eggs and those in coastal marine sediments. Based on limited toxicological information, the predicted no effect concentrations (PNECs) of Hg and Se in the bird eggs were estimated as 0.5 to 1.8 and 0.90 to 1.45 mug g(-1), dry wt., respectively. Hazard quotients (HQs) indicated that Hg and Se might pose some risk to certain waterbirds in Hong Kong. PMID- 15749546 TI - Equilibrium partitioning of (14)C-benzo(a)pyrene and (14)C-benazolin between fractionated phases from an arable topsoil. AB - Equilibrium partitioning of hydrophobic (14)C-benzo(a)pyrene and hydrophilic (14)C-benazolin between fractionated phases from an arable topsoil of Merzenhausen (Germany) was investigated. Topsoil samples were collected from lysimeters which were incubated with different residual crops. A physical soil fractionation based on sedimentation and centrifugation steps was performed after water extraction. Four soil phases were obtained designated sediment (SED) phase (>20 microm), microaggregate (MA) phase (2 microm-20 microm), colloid (COL) phase (<2 microm) and electrolyte (EL) phase. The distribution of (14)C-benzo(a)pyrene, (14)C-benazolin and organic carbon between the soil phases was established. Enrichment factors for the two chemicals and organic carbon are higher in the COL and MA phases than in the SED phase. The distribution constant K(d) of chemicals for Merzenhausen topsoil was calculated according to two-phase or three-phase partitioning models. The three-phase partitioning model presumes the contribution of the organic carbon for the binding of chemicals. A log K(oc) of 5.55 can be calculated in the case of (14)C-benzo(a)pyrene, which is typical of the hydrophobic association with the soil organic carbon. In the case of (14)C benazolin, much higher K(d) and K(oc) values were calculated than found with parent molecules after short-time experiments. Long-term aging processes must be considered. Specific effects on the chemical distribution due to the different crop residues were not detected. PMID- 15749547 TI - The use of autecological and environmental parameters for establishing the status of lichen vegetation in a baseline study for a long-term monitoring survey. AB - In 1997 the ecological characteristics of the epiphytic species (83 lichens and two algae) of an urban area (Grenoble, France) were determined. Seven autecological indices were used to characterize the lichen ecology: illumination index, humidity index, pH of bark, nutrient status of substratum, ecological index of IAP and frequency. Six clusters (A1-A6) were defined using cluster analysis and principal component analysis. Seven environmental parameters characterizing the stations and the lichen releves were also used: elevation, parameters of artificiality (urbanization, traffic and local land use), IAP, and the percentage of nitrophytic and acidophytic species. Six clusters (B1-B6) were defined using cluster analysis and canonical correspondence analysis. Four clusters (C1-C4) were finally defined using an empirical integrated method combining the autecological and environmental parameters. This final clustering which established the status of the lichen vegetation in 1997 can be reliably used as a baseline study to effectively monitor environmental changes in this urban area. PMID- 15749548 TI - Soil remediation: humic acids as natural surfactants in the washings of highly contaminated soils. AB - The remediation of the highly contaminated site around the former chemical plant of ACNA (near Savona) in Northern Italy is a top priority in Italy. The aim of the present work was to contribute in finding innovative and environmental friendly technology to remediate soils from the ACNA contaminated site. Two soils sampled from the ACNA site (A and B), differing in texture and amount and type of organic contaminants, were subjected to soil washings by comparing the removal efficiency of water, two synthetic surfactants, sodium dodecylsulphate (SDS) and Triton X-100 (TX100), and a solution of a natural surfactant, a humic acid (HA) at its critical micelle concentration (CMC). The extraction of pollutants by sonication and soxhlet was conducted before and after the soil washings. Soil A was richer in polycyclic aromatic hydrocarbons, whereas soil B had a larger content of thiophenes. Sonication resulted more analytically efficient in the fine-textured soil B. The coarse-textured soil A was extracted with a general equal efficiency also by soxhlet. Clean-up by water was unable to exhaustively remove contaminants from the two soils, whereas all the organic surfactants revealed very similar efficiencies (up to 90%) in the removal of the contaminants from the soils. Hence, the use of solutions of natural HAs appears as a better choice for soil washings of highly polluted soils due to their additional capacity to promote microbial activity, in contrast to synthetic surfactants, for a further natural attenuation in washed soils. PMID- 15749549 TI - Liver echogenicity: relation to systemic blood pressure and other components of the metabolic syndrome. AB - We studied the impact of liver echogenicity among other potential predictors of systemic blood pressure (BP) and the metabolic syndrome. 38 persons (32 males, six females, aged 29 to 66) had their liver echogenicities scored, BPs measured and standard serum laboratory tests studied. There was a significant correlation between both systolic (r=0.438, p=0.006) and diastolic (r=0.498, p=0.001) BP and liver echogenicity. Liver echogenicity was the strongest predictor for systolic BP and the second strongest (after body mass index, BMI) for diastolic BP. Body height may modify the relation between liver echogenicity and systolic BP. Liver echogenicity also correlated significantly with BMI (r=0.527, p=0.001), serum triglycerides (r=0.472, p=0.003) and, to a lesser degree, with serum total cholesterol (r=0.305, p=0.066). Incidentally found increased liver echogenicity should alert the US performer and the clinician reading the report on the possibility of elevated BP or other features of the metabolic syndrome. PMID- 15749550 TI - Staging bladder carcinoma by three-dimensional ultrasound rendering. AB - Organ-confined staging for bladder cancer has major impact on further treatment. Most imaging techniques for this purpose are insufficient. We, therefore, assessed the value and the limitations of a new diagnostic tool, the 3-D ultrasound (US) rendering, to distinguish invasive from noninvasive bladder cancers. A total of 63 patients underwent 3-D US of the bladder before transurethral resection or radical cystectomy. The US findings were compared with the pathologic stages of the transurethral resection material or the cystectomy specimens. Superficial (pTa) carcinomas were correctly staged in 66% by 3-D US. Lamina propria infiltrating (pT1) were correctly staged in 83% and the quota of correct staging of infiltrating carcinomas (>pT1) by 3-D rendering was 100%. The overall accuracy was 79%. Three-dimensional US rendering is most valuable to discriminate between superficial stages pT1. This new technique might improve staging of bladder cancer. PMID- 15749551 TI - Identification of the layers of the bladder wall on high-frequency endoluminal ultrasonography by a needle puncture experiment. AB - The purpose of this study was to identify the layers of the bladder wall on high frequency endoluminal ultrasonography (ELUS). We performed a needle puncture experiment using five normal pig bladders. The histologic layer structure was compared with the ELUS images obtained using a 30-MHz miniature transducer. The bladder wall was depicted as five layers by ELUS and these corresponded to the histologic layers. The first layer (hyperechoic) was a margin echo that included the epithelium and the upper part of the lamina propria. The second layer (hypoechoic) corresponded to the lamina propria and the third layer (hyperechoic) was a margin echo that included the upper part of the muscle layer. The fourth layer (hypoechoic) corresponded to the muscle layer, and the fifth layer (hyperechoic) was a margin echo that included the upper part of the adventitia. PMID- 15749552 TI - Multiplanar transcranial ultrasound imaging: standards, landmarks and correlation with magnetic resonance imaging. AB - The purpose of this study was to define a standardized multiplanar approach for transcranial ultrasound (US) imaging of brain parenchyma based on matched data from 3-D US and 3-D magnetic resonance imaging (MRI). The potential and limitations of multiple insonation planes in transverse and coronal orientation were evaluated for the visualization of intracranial landmarks in 60 healthy individuals (18 to 83 years old, mean 41.4 years) with sufficient temporal bone windows. Landmarks regularly visualized even in moderate sonographic conditions with identification rates of >75% were mesencephalon, pons, third ventricle, lateral ventricles, falx, thalamus, basal ganglia, pineal gland and temporal lobe. Identification of medulla oblongata, fourth ventricle, cerebellar structures, hippocampus, insula, frontal, parietal and occipital lobes was more difficult (<75%). We hypothesize that multiplanar transcranial US images, with standardized specification of tilt angles and orientation, not only allow comparison with other neuroimaging modalities, but may also provide a more objective framework for US monitoring of cerebral disease than freehand scanning. PMID- 15749553 TI - Acceleration time in the fetal middle cerebral artery during the second half of pregnancy. AB - We made a longitudinal ultrasonographic study of 33 healthy fetuses from 22 to 38 weeks of gestation to determine the acceleration time in the middle cerebral artery (ATMCA) and its relationship with fetal cardiac output. Doppler ultrasound (US) was performed by a single observer always using the same apparatus. The Doppler tracing was obtained with a sample volume of 1 mm; placement was on the anterior middle cerebral artery, as close as possible to the skullcap. The insonation angle was kept between 5 degrees and 19 degrees and the wall filter was adjusted to a frequency of 50 to 100 Hz. The ATMCA was 40 ms at 22 and 26 weeks, 50 ms at 30 and 34 weeks and 60 ms at the 38th week. There was a significant positive correlation between gestational age and ATMCA values (r=0.45, p<0.001). The ATMCA values became significantly higher than the initial values at the 30th week of gestation (p<0.05). The ATMCA values for normal fetuses could be compared with those obtained during high-risk pregnancies; this would be useful for the detection of fetal cardiac anomalies. PMID- 15749554 TI - Computer analysis of three-dimensional power angiography images of foetal cerebral, lung and placental circulation in normal and high-risk pregnancy. AB - Three-dimensional (3-D) ultrasound (US) has greatly improved evaluation of organ circulation. The aim of this study was to explore the possible use of this new technique in normal and high-risk pregnancies. Fetal brain, lung and placenta 3-D power Doppler signal intensity were recorded in 115 normal singleton pregnancies (24 to 42 weeks gestation) and in 67 high-risk pregnancies. Mean image pixel signal intensity was calculated for each organ and a brain-lung ratio. In normal pregnancy, placental and lung signal intensity increased until 33, with a rapid decrease after 38, weeks of gestation. Fetal cerebral signal intensity increased with gestational age. Placental and fetal lung signal intensity was significantly lower in high-risk pregnancies than in the control group, with increased fetal brain and brain-lung ratios. The present results suggest a reduction of placental perfusion after 38 weeks of gestation in normal pregnancy, with redistribution of fetal circulation. Lung signal intensity increased abruptly at 32 weeks of gestation, which might reflect lung maturity. The new method showed signs of centralization of fetal circulation at the end of gestation. The results might suggest a possible clinical use for fetal surveillance in high-risk pregnancies. PMID- 15749555 TI - Development and validation of an in vivo analysis tool to identify changes in carotid plaque tissue types in serial 3-D ultrasound scans. AB - We have developed a three-dimensional (3-D) B-mode acquisition system suitable for imaging carotid plaques in vivo. A texture classification system using 157 statistical and textural algorithms, previously developed in our laboratory and shown to predict the contents of in vitro carotid plaques, was applied to in vivo 3-D image sets obtained from patients with both symptomatic and asymptomatic carotid artery plaques. Delineation of plaque boundaries is more difficult using in vivo images than in vitro images of excised plaques embedded in agar. This study has examined inter- and intraobserver variability studies to assess the degree of selectivity of the plaque region-of-interest (ROI) and assess the degree of repeatability for potential use in comparing serial scans. An interobserver limit of agreement of +/-12.9% and an intraobserver limit of repeatability of <2% were obtained. These results show that the plaque ROI selection is subjective, but is repeatable within acceptable limits. PMID- 15749556 TI - Diclofenac phonophoresis in human volunteers. AB - A quantitative study of sodium diclofenac (Voltaren Emulgel, Novartis) phonophoresis was undertaken in humans. Fourteen healthy human volunteers were submitted to ultrasound irradiation on two 225-cm2 areas on the dorsum (group A), followed by the application of the medication gel, and the plasma diclofenac mass was measured at 1, 2 and 3 h later by high performance liquid chromatography. The same procedure was repeated one month later with the same volunteers but with the ultrasound equipment switched off for the control group (group B). The plasma diclofenac mass was significantly higher in group A than in group B at 1 h (0.0987 microg/mL as opposed to 0.0389 microg/mL; p=0.01) and 2 h (0.0724 microg/mL as opposed to 0.0529 microg/mL; p=0.01), but not at 3 h (0.0864 microg/mL as opposed to 0.0683 microg/mL; p=0.16). The authors conclude that previously applied therapeutic ultrasound irradiation enhances the percutaneous penetration of the topical diclofenac gel, although the mechanism remains unclear. PMID- 15749557 TI - Comparative study of the efficacy of the topical application of hydrocortisone, therapeutic ultrasound and phonophoresis on the tissue repair process in rat tendons. AB - The purpose of this study was to compare the treatment efficacy of topical application of hydrocortisone, therapeutic ultrasound (US) and phonophoresis on the rat's Achilles tendon (tendo calcaneus) repair process after tenotomy. The two treated groups with US were made in a pulsed mode. The irradiation of US was performed at a frequency of 1 MHz and an intensity of 0.5 W/cm2 (SATA), for 5 min each session. The tendons were analyzed using the polarized light microscopy. The results showed that the treated group with the topical application of hydrocortisone has not been delivered transdermally and that the molecule of collagen responds to the ultrasonic stimulation. The treatment with phonophoresis was the more efficient method. These findings allow us to conclude that the US stimulates the acceleration of tissue repair processes and induces the transdermal delivery of hydrocortisone in a therapeutic concentration on the tendon. PMID- 15749558 TI - Association of automated and human observer lesion detecting ability using phantoms. AB - A set of tissue-mimicking phantoms containing spherical negative contrast simulated lesions was employed to associate an automated method for determining detectability with human observers. Six alternative methods for computing the lesion signal-to-noise ratio (LSNR) were employed for quantifying automated detecting ability. The six methods differ regarding effective lesion area and whether or not gradients in local mean background echo levels were accounted for. The two-alternative-forced-choice (TAFC) technique was used to associate detecting ability of human observers with LSNR values. Although the six methods gave similar results, one method exhibited the least dependency on lesion diameter and is recommended; that method accounts for gradients in local mean background echo levels and employs an effective sphere area of 2/pi times the projected sphere area. A reasonable LSNR detection threshold value of -2.0 was found to apply for nominal transducer frequencies from 4 through 6 MHz and for lesion diameters from 2 through 5 mm. This result allows rapid human-observer calibrated automated determination of the depth range of detectability as a function of sphere diameter and contrast. PMID- 15749559 TI - Texture analysis in digitally-acquired echocardiographic images: the effect of JPEG compression and video storage. AB - The analysis of texture in video-stored echocardiographic images is an established method to characterize myocardial pathologies. We investigated whether or not texture parameters calculated from video-stored images and those derived from the joint photographic expert group (JPEG) format compressed data are equivalent to those calculated from uncompressed digital images. Texture parameters were calculated using uncompressed digital data, images stored on videotape, and three forms of compressed digital data (baseline JPEG, JPEG 2000 and lossless JPEG 2000). Video storage heavily affected most texture parameters. Although first-order texture parameters derived from JPEG-compressed images were generally equivalent to those derived from the uncompressed data, several second order parameters differed significantly. We conclude that texture of video-stored images is not comparable to that of digitally-stored images and that JPEG compression changes important second-order texture parameters. This observation should be taken into account when analyzing texture of modern image data (uncompressed or compressed) and comparing the results with earlier studies utilizing video-stored data. PMID- 15749560 TI - Ultrasonic vibration dectection with wavelets: preliminary results. AB - Several arterial disorders are known to cause systolic audio vibrations in tissue: they include stenoses, vasospasm, aneurysms, bleeds and arteriovenous fistulas. High-amplitude vibrations can be discovered with conventional Doppler ultrasound (US) instruments; however, differentiating brief, low-amplitude vibrations from other nonstationary echo sources is difficult. Further, characterizing the frequency and amplitude of vibrations is not feasible with conventional Doppler US. The automated detection and estimation of both the frequency and amplitude of vibrations with durations less than 100 ms and amplitudes of a micrometer or less have remained a signal-processing challenge. These vibrations may be associated with both nonstationary colored noise and strong low-frequency clutter. The normalized continuous Morlet wavelet power spectrum analysis of quadrature Doppler echoes, followed by a binary hypothesis test for noise, results in simulated detection rates above 99.9%, with 0.1% false alarms for signal-on signal-to-noise ratios (SNRs) as low as one. Two clinical examples are included. PMID- 15749561 TI - Pressure-dependent attenuation with microbubbles at low mechanical index. AB - It has previously been shown that the attenuation of ultrasound (US) by microbubble contrast agents is dependent on acoustic pressure (Chen et al. 2002). Although previous studies have modelled the pressure-dependence of attenuation in single bubbles, this paper investigates this subject by considering a bulk volume of bubbles together with other linear attenuators. Specifically, a new pressure dependent attenuation model for an inhomogeneous volume of attenuators is proposed. In this model, the effect of the attenuation on US propagation is considered. The model was validated using experimental measurements on the US contrast agent Sonovue. The results indicate, at low acoustic pressures, a linear relationship between the attenuation of Sonovue, measured in dB, and the insonating acoustic pressure. PMID- 15749562 TI - Erythrocytes, as well as microbubble contrast agents, are important factors in improving thermal and therapeutic effects of high-intensity focused ultrasound. AB - Erythrocytes, as well as microbubble contrast agents, are important factors in improving thermal effect of high-intensity focused ultrasound (US), or HIFU, and increasing the coagulation volume produced by HIFU irradiation. In vitro experiments used human plasma with various concentrations of human erythrocytes in combination with or without Levovist. In vivo experiments used eight Japan white rabbits with three degrees of anemia. Using a 2.17-MHz transducer, HIFU was applied for 60 s, and the temperature rise and the volume of coagulation necrosis was evaluated. There was a significant correlation between the HIFU-induced temperature rise and hematocrit, with a correlation coefficient of 0.998 (p=0.0001). Although the temperature rise was smaller at low hematocrit, it was significantly increased by adding Levovist in the suspension (p<0.01). The mean volume of coagulation necrosis was significantly greater in the rabbits with higher hematocrits (p<0.01), and that in the moderate anemia group was significantly increased by using Levovist (p<0.01). PMID- 15749563 TI - High-speed optical observations of contrast agent destruction. AB - Ultrasound contrast agents are now available since a few years and used for diagnostic purposes. Improved diagnostic decisions have been made possible with new imaging methods that are mainly based on the nonlinear properties of gas microbubbles. Since it is well known that contrast agents are destroyed by ultrasound when the acoustic pressure exceeds a threshold, extremely low acoustic pressures were applied to achieve enhanced contrast image quality. However, destruction of contrast microbubbles is not necessarily undesirable, since it is beneficial in, for example, destruction/reperfusion imaging and recently in drug delivery. We investigate in this experimental study the destruction dynamics of a contrast agent consisting of nitrogen bubbles encapsulated in a double polymer/albumin wall shell. This is accomplished using an ultrafast camera Brandaris that operates at a frame rate of 25 MHz and records 128 frames. The measurements were performed with an ultrasound sine burst of 10 cycles at 1.7 MHz. Different acoustic pressures were applied and various microsphere sizes were examined. The results show three different zones depending on the applied pressure and bubble size: these are nondestruction zone, transient zone and destruction zone. The nondestruction zone is reached for either very small microspheres or low mechanical indices (MI) (<0.3). In the destruction zone lie either large microspheres (5 microm or higher) even when irradiated at low MIs or small microspheres (<5 microm) when the MI is above 0.6. The optical observations revealed that the destruction of the microspheres is characterized by shell rupture and gas release. The release of the gas gives rise to new free microbubble that lasts for a few milliseconds and then disappears due to dissolution. In the transient zone, the microspheres are mainly compressed in the first few cycles but no expansion is induced. After intense compressions, the shell fissures and gas escapes in the last cycles of the burst or during a second burst depending on the initial size and MI. These optical recordings are important to investigate contrast bubble destruction and can help in amplifying or minimizing this process. Indeed, bubble disruption remains the basis of most current sensitive methods for detecting perfusion with contrast agents and is an essential component of perfusion quantification with microbubbles, in addition to drug delivery applications and pressure measurements. PMID- 15749564 TI - Ultrasonic temperature imaging for guiding focused ultrasound surgery: effect of angle between imaging beam and therapy beam. AB - Ultrasonic temperature imaging is a promising technique for guiding focused ultrasound surgery (FUS). The FUS system is run at an initial, nonablative intensity and a diagnostic transducer images the heat-induced echo strain, which is proportional to the temperature rise. The echo strain image portrays an elliptical "hot spot" corresponding to the focal region of the therapy transducer. It is anticipated that such images will be used to predict the location of the thermal lesion that would be produced at an ablative intensity. We demonstrated in vitro that heat-induced echo strain images can visualize a spatial peak temperature rise of <2 degrees C (starting at room temperature). However, the imaging beam was perpendicular to the treatment beam in these experiments, whereas the most convenient approach in vivo would be to mount the imaging probe within the housing of the therapy transducer such that the two beams are coaxial. A previous simulation experiment predicted that echo strain images would be noisier for the coaxial configuration because sharp lateral gradients in axial displacement cause increased RF signal decorrelation within the beam width. The aim of the current study was to verify this prediction in vitro. We found, that for a temperature rise of approximately 4 degrees C, the mean contrast-to-noise ratio for coaxial and perpendicular echo strain images was 0.37 (+/-0.24) and 2.00 (+/-0.72) respectively. Furthermore, the decorrelation noise seen in the coaxial images obscured the posterior axial border of the hot spot. We conclude that the coaxial configuration will be useful for localizing the hot spot in the lateral direction. However, it may not be able to depict the axial extent of the hot spot or to portray a parameter that is directly related to temperature rise. PMID- 15749565 TI - Monitoring stiffness changes in lesions after radiofrequency ablation at different temperatures and durations of ablation. AB - The variations in the stiffness or stiffness contrast of lesions resulting from radiofrequency (RF) ablation of canine liver tissue at different temperatures and for different ablation durations at a specified temperature are analyzed. Tissue stiffness, in general, increases with temperature; however, an anomaly exists around 80 degrees C, where the stiffness of the lesion is lower than that of the lesion ablated at 70 degrees C. On the other hand, the stiffness increases monotonically with the duration of ablation. Plots illustrating the ratio of mean strains in normal canine liver tissue to mean strains in ablated thermal lesions demonstrate the variation in the stiffness contrast of the thermal lesions. The contrast-to-noise ratio (CNRe) of the lesions, which serves as an indicator of the detectability of the lesions under the different experimental imaging conditions described above, is also presented. The results presented in this paper show that the elastographic depiction of stiffer thermal lesions is better, in terms of the CNRe parameter. An important criterion in the elastographic depiction of RF-ablated regions of tissue is the trade-off between ablation temperature and duration of ablation. Tissue necrosis can occur either by ablating tissue to high temperatures for short durations or to lower temperatures for longer durations. In this paper, we attempt to characterize the elastographic depiction of thermal lesions under these different experimental conditions. This paper provides results that may be utilized by practitioners of RF ablation to decide the ablation temperature and duration, on the basis of the strain images of normal liver tissue and ablated thermal lesions discussed in this paper. PMID- 15749566 TI - Physical enviroment of 2-D animal cell aggregates formed in a short pathlength ultrasound standing wave trap. AB - 2-D mammalian cell aggregates can be formed and levitated in a 1.5 MHz single half wavelength ultrasound standing wave trap. The physical environment of cells in such a trap has been examined. Attention was paid to parameters such as temperature, acoustic streaming, cavitation and intercellular forces. The extent to which these factors might be intrusive to a neural cell aggregate levitated in the trap was evaluated. Neural cells were exposed to ultrasound at a pressure amplitude of 0.54 MPa for 30 s; a small aggregate had been formed at the center of the trap. The pressure amplitude was then decreased to 0.27 MPa for 2 min, at which level the aggregation process continued at a slower rate. The pressure amplitude was then decreased to 0.06 MPa for 1 h. Temperature measurements that were conducted in situ with a 200 microm thermocouple over a 30 min period showed that the maximum temperature rise was less than 0.5 K. Acoustic streaming was measured by the particle image velocimetry method (PIV). It was shown that the hydrodynamic stress imposed on cells by acoustic streaming is less than that imposed by gentle preparative centrifugation procedures. Acoustic spectrum analysis showed that cavitation activity does not occur in the cell suspensions sonicated at the above pressures. White noise was detected only at a pressure amplitude of 1.96 MPa. Finally, it was shown that the attractive acoustic force between ultrasonically agglomerated cells is small compared with the normal attractive van der Waals force that operates at close cell surface separations. It is concluded that the standing wave trap operates only to concentrate cells locally, as in tissue, and does not modify the in vitro expression of surface receptor interactions. PMID- 15749567 TI - The effects of low-intensity ultrasound on growing bone after sciatic neurectomy. AB - The purpose of this study was to investigate the therapeutic effects of low intensity ultrasound (US) on the sciatic-neurectomy-induced bone mass decrement in growing rats. A total of 20 male Sprague-Dawley rats (166.7+/-11.3 g) underwent right leg sciatic neurectomy. They were randomly assigned into two groups, US treatment group (UST) and control group (CON). The rats receiving US treatment were treated with a 125 mW/cm2 continuous low-intensity US stimulation for 15 min/day on the lateral site of the right leg. The control rats did not receive any US treatment. All the animals were euthanized after 4-week US treatment. Both the original data and bilateral difference ratio of femur or tibia weight, histomorphometry data and bone densitometry data showed that the sciatic neurectomy obviously reduced bone mass in the operated limbs of both groups. However, the continuous low-intensity US treatment did not ameliorate the neurectomy-induced loss of bone mass. Thus, the low-intensity US generated micromechanical strains might not induce enough bone formation activity to reverse the bone loss in this model of intact bones. PMID- 15749568 TI - The effect of size on the acoustic response of polymer-shelled contrast agents. AB - In this technical note, we study three polymer-shelled microbubble contrast agents manufactured by POINT Biomedical Corporation that have identical shell composition and mean volumetric diameters of 0.74 microm, 0.91 microm and 1.33 microm. We investigate the effect of agent size on the amplitude, frequency and probability of acoustic echoes received in response to five-cycle, 2.25-MHz pulses of varying pressure. We find that the amplitude and frequency response from the three agents is not significantly different. However, significant differences among the agents do exist in the probability of response to acoustic interrogation: at a pressure of 1.06 MPa, an echo from the 1.33 microm agent is 5 times as likely as an echo from the 0.91 microm agent and 18 times as likely as an echo from the 0.74 microm agent. We hypothesize that there exists an effective pressure-dependent threshold diameter above which single polymer-shelled agents respond to acoustic interrogation. PMID- 15749569 TI - Gender differences in the effects of family adversity on the risk of onset of DSM III-R social phobia. AB - Utilizing a random probability sample of Canadian residents aged 15-64 (n = 8116), this study assessed gender differences in the onset of social phobia and the moderating influence of gender on indicators of childhood family adversity hypothesized to increase the risk of developing the disorder. Results revealed statistically significant "gender by family adversity" interactions that varied by disorder sub-type. Among males, absence of a parent or other adult close confidant during childhood was associated with an elevated risk of developing social phobia (all diagnosed cases and the non-generalized sub-type). Risk factors unique to females included parental conflict while growing up (all diagnosed cases), childhood physical abuse by a father figure (generalized sub type), and maternal mania (non-generalized sub-type). Results highlight the importance of distinguishing between social phobia sub-types in gender-based research as well as the use of family adversity measures that capture the parenting behaviors and mental health status of both parents. PMID- 15749570 TI - The phenomenology of non-clinical panic in older adults in comparison to younger adults. AB - The prevalence and nature of panic symptoms in the elderly are poorly understood. In a replication and extension of Deer and Calamari [J. Behav. Ther. Exp. Psychiatry 29 (1998) 303], the current study sought to determine which symptoms of anxiety best differentiated elderly endorsers of panic from elderly who do not panic, and to compare these results to a reference younger sample. Based on a sample of 302 community-dwelling older adults and a comparison group of 275 younger adults, non-clinical panic (NCP) was endorsed in 26.2% of older adults, which is less frequent than the younger adults (42.7%). Although older and younger adults were similar in terms of clinical features of panic, health factors interacted significantly with panic symptoms in the elderly. When controlling for health, cognitive symptoms of anxiety were stronger predictors of panic in older adults than were physiological symptoms. Clinical implications for the assessment of panic in older adults are discussed. PMID- 15749571 TI - The effect of fear on paralinguistic aspects of speech in patients with panic disorder with agoraphobia. AB - The present study investigated the effect of fear on paralinguistic aspects of speech in patients suffering from panic disorder with agoraphobia (N = 25). An experiment was conducted that comprised two modules: Autobiographical Talking and Script Talking. Each module consisted of two emotional conditions: Fearful and Happy. Speech was recorded digitally and analyzed using PRAAT, a computer program designed to extract paralinguistic measures from digitally recorded spoken sound. In addition to subjective fear, several speech characteristics were measured as a reflection of psychophysiology: rate of speech, mean pitch and pitch variability. Results show that in Autobiographical Talking speech was slower, had a lower pitch, and a lower pitch variability than in Script Talking. Pitch variability was lower in Fearful than in Happy speech. The findings indicate that paralinguistic aspects of speech, especially pitch variability, are promising measures to gain information about fear processing during the recollection of autobiographical memories. PMID- 15749572 TI - Using facial expressions as CSs and fearsome and disgusting pictures as UCSs: affective responding and evaluative learning of fear and disgust in blood injection-injury phobia. AB - Two experiments examine use of an evaluative conditioning (EC) paradigm in the acquisition of fear and disgust responding to neutral facial expressions. In Experiment 1, 60 participants were randomly assigned to one of three evaluative learning conditions in which neutral facial expressions were paired with fearsome, disgusting, or neutral pictures. No statistically significant differences were detected between the three conditions. However, significant differences emerged within subjects as post-exposure of fear and disgust ratings were higher among expressions that were paired with pictorial stimuli. Experiment 2 sought to examine if an analogue sample of BII phobics would be more susceptible than nonphobic controls to fear and disgust EC utilizing a similar experimental design, given the co-occurrence of fear and disgust in BII-phobic responding. Results failed to demonstrate an EC effect specific to the analogue phobic group, although both groups showed an evaluative shift toward disgust for those facial expressions paired with BII-relevant pictures. Consistent with previous findings, examination of picture rating data suggested that analogue BII phobics rated the BII pictures as significantly more disgusting than fearful. The role of EC processes and a priori expectancy biases in the associative learning of disgust in BII phobia is discussed. PMID- 15749573 TI - Level of emotional awareness as a differentiating variable between individuals with and without generalized anxiety disorder. AB - Using Mennin, Heimberg, Turk, and Fresco's [Emotion regulation deficits as a key feature of generalized anxiety disorder: Testing a theoretical model, submitted for publication] conceptualization of generalized anxiety disorder (GAD) as a syndrome involving emotion dysregulation and an overuse of cognitive control strategies, this study sought to differentiate individuals with GAD from controls by offering differences in emotional awareness as one of the central distinctions between these groups. This study employs the Levels of Emotional Awareness Scale (LEAS) [Lane et al., 1990Lane, R. D., Quinlan, D. M., Schwartz, G. E., Walker, P. A., & Zeitlin, S. B. (1990). The Levels of Emotional Awareness Scale: a cognitive developmental measure of emotion. Journal of Personality Assessment, 55, 124-134] a rater-coded measure, to assess level of emotional awareness, a methodological improvement over previous tests of the model, which relied upon self-report. Individuals with GAD scored significantly higher than controls on emotional awareness. These findings are discussed in light of the theoretical implications for GAD. PMID- 15749574 TI - The Yale-Brown Obsessive Compulsive Scale: factor analysis, construct validity, and suggestions for refinement. AB - The Yale-Brown Obsessive Compulsive Scale (Y-BOCS) is widely acknowledged as the gold standard measure of obsessive-compulsive disorder (OCD) symptom severity. Despite its popularity, a number of questions remain regarding the Y-BOCS' psychometric properties including: (a) whether obsessional and compulsive symptoms contribute independently to global OCD severity and (b) whether the Y BOCS subscales are valid with respect to other measures of OCD. We examined these issues in a sample of 100 patients with a diagnosis of OCD. While our confirmatory factor analyses failed to reproduce any previously reported models of the Y-BOCS factor structure, exploratory factor analysis indicated a two factor solution that assessed symptom severity (i.e., time, distress, and interference from obsessions and compulsions) as separate from resistance and control of obsessions and compulsions. In contrast to the Resistance/Control Subscale, the Severity Subscale demonstrated good psychometric properties and construct validity. Based on our findings we recommend revisions to scoring the Y BOCS. PMID- 15749575 TI - Induced arousal and reexperiencing in acute stress disorder. AB - This study investigated the relationship of hyperarousal and intrusive symptoms in acute stress disorder (ASD). Civilian trauma survivors with ASD (n = 18) and without ASD (n = 14) completed a hyperventilation provocation test (HVPT) and then completed the Physical Reactions Scale. All participants provided a narrative describing their hyperventilation experience that was audiotaped and independently coded. Individuals with ASD reported greater numbers of intrusive memories and reported greater distress than non-ASD participants. More ASD than non-ASD participants experienced a flashback-type reaction during the hyperventilation. Intrusive symptoms were significantly correlated with elevated arousal following the HVPT. The findings provide evidence that reexperiencing is directly associated with elevated states of arousal. PMID- 15749578 TI - Gastrointestinal disease in adolescents. PMID- 15749576 TI - Sleep paralysis in adults reporting repressed, recovered, or continuous memories of childhood sexual abuse. AB - Sleep paralysis typically occurs as individuals awaken from rapid eye movement sleep before motor paralysis wanes. Many episodes are accompanied by tactile and visual hallucinations, often of threatening intruders in the bedroom. Pendergrast [Victims of Memory: Incest Accusations and Shattered Lives, HarperCollins, London, 1996] proposed that individuals who report repressed or recovered memories of childhood sexual abuse (CSA) may misinterpret episodes of sleep paralysis as reemerging fragments of dissociated ("repressed") memories of CSA. To investigate this issue, we administered a sleep paralysis questionnaire to people reporting either repressed (n = 18), recovered (n = 14), or continuous (n = 36) memories of CSA, or to a control group reporting no history of CSA (n = 16). The prevalence of sleep paralysis was: repressed memory group (44%), recovered memory group (43%), continuous memory group (47%), and control group (13%). Among the six individuals in the recovered memory group who had experienced sleep paralysis, one interpreted it as related to sexual abuse (i.e., a rate of 17%). All other participants who had reported sleep paralysis embraced other interpretations (e.g., saw a ghost). Dissociation and depressive symptoms were more common among those who had experienced sleep paralysis than among those who denied having experienced it. PMID- 15749579 TI - Sports medicine and the adolescent female. PMID- 15749580 TI - Disclosure of date/acquaintance rape: who reports and when. AB - OBJECTIVE: To estimates rates and correlates of disclosure of date/acquaintance rape or attempted rape and verbally coercive sex among a diverse sample of adolescent and young adult females. DESIGN: Secondary data analysis of cross sectional data. SETTING: Urban adolescent healthcare facility. PARTICIPANTS: Adolescents who were identified as having experienced rape/attempted rape (n = 86) or verbally coerced sex (n = 68) in the last 12 months from study examining sexual violence. MAIN OUTCOME MEASURES: Disclosure of forced sex (logistic regression) and the timing of disclosure (survival analysis). INTERVENTIONS: None. RESULTS: Almost 60% of victims who experienced rape/attempted rape disclosed this information to one or more individuals, whereas only 47% of those who experienced verbally coerced sex told another person. Multivariate analyses found that drinking by the partner (AOR = 4.6) and shorter dating history (AOR = 6.3) were associated with disclosure of rape/attempted rape; timing of this disclosure was facilitated by Caucasian ethnicity (RR = 3.5), having a dating partner who drank > or = 1 drinks (RR = 2.5), and the perpetrator being someone other than the victim's boyfriend or partner (RR = 2.5). With regards to verbally coerced sex, reporting no pressure to use alcohol (AOR = 10.7) was the only factor associated with disclosure. No significant predictors of timing to disclosure were detected for this type of victimization. CONCLUSIONS: Perpetrator's alcohol use and a shorter dating history are important variables associated with disclosure of rape/attempted rape as well as timing to disclosure. Factors affecting the disclosure of verbally coerced sex and the latency associated with disclosure are less well defined. PMID- 15749581 TI - Changes in young women's awareness, attitudes, and perceived barriers to using emergency contraception. AB - BACKGROUND: In a 1996 survey, most young women ages 13-20 years from an urban, hospital-based clinic and a drug treatment center had inadequate awareness of emergency contraception (EC), and perceived several barriers to its use. Since that time, the FDA has approved two products for EC, media coverage has increased, and physicians have provided more counseling about EC. PURPOSE: The purpose of this study is to compare the awareness, attitudes, and perceived barriers to using EC among a sample of young women from 1996 with a different sample of women from 2002. METHODS: We recruited 139 young women (mean age 16.7 +/- 1.8 yrs) from the same adolescent clinic and drug treatment center as the 1996 sample. They had similar demographic characteristics, with the majority (63%) being African-American or multi-ethnic; 85% had ever been sexually active. They were interviewed using a questionnaire about their sexual and contraceptive history as well as their knowledge of and experience with EC. They then watched a 4(1/2) minute video and received a 5-minute didactic review of EC. Following the educational intervention, participants' knowledge, attitudes, and perceived barriers to using EC were assessed. The questionnaire used to guide the interviews was nearly identical to that used in 1996. RESULTS: Between 1996 and 2002, the percentage of participants reporting that they had ever heard of EC grew (44% vs. 73%, P < 0.001), as well as the percentage reporting that they had ever used EC (4% vs. 13%, P = 0.02). Of those participants who had ever heard of EC, fewer 1996 participants knew where to obtain it compared to 2002 participants (78% vs. 95%, P = 0.002) and fewer 1996 participants knew the correct time limits for use (20% vs. 51%, P < 0.001). The above data were collected prior to a didactic review session about EC. After receiving information about EC, the percentage of participants reporting a positive attitude toward EC grew between 1996 and 2002 (72% vs. 96%, P < 0.001). Young women also had fewer concerns about safety and side effects in 2002. The 1996 participants were more likely to report barriers to using EC compared to the 2002 participants. In 1996, EC side effects and impact on fertility were the most commonly perceived barriers to EC use. However, in 2002 the frequency of all reported barriers decreased and cost had become the number one perceived barrier. CONCLUSION: Since 1996, young women at an urban hospital-based adolescent clinic and drug treatment center increased their awareness, use, and positive attitudes towards EC, as well as decreased their perceived barriers to using EC. Educational interventions that focus on improving knowledge among younger adolescents, specifically about correct time limits and identifying ways to find affordable EC, will address the most common knowledge deficits and perceived barriers to EC use among adolescents. PMID- 15749582 TI - Are adolescent girls with Chlamydia infection notifying their partners? AB - OBJECTIVES: (1) To determine the proportion of inner-city adolescent girls diagnosed with chlamydial cervicitis who notify their sex partners; (2) to examine girls' attitudes and perceptions about partner notification and treatment; and (3) to assess whether or not girls knew if their partners were treated for chlamydia infection. DESIGN/METHODS: Adolescent girls who had a positive DNA hybridization test for chlamydial cervicitis from March 2000 to May 2002 completed a 37-item self-administered survey assessing sexual behavior and partner notification, as well as the Rosenberg self esteem scale. Subjects completed the survey 1-3 months after the diagnosis of chlamydia infection. RESULTS: Fifty-five adolescent girls (46% Hispanic, 36% African American) aged 13 21 years (mean 18.3 years) completed the survey. The median age at first intercourse was 14 (SD = 1.6); median number of lifetime sex partners was 4. Forty-one subjects (75%) notified their sex partners. The most common reasons for partner notification were"I did not want my sex partner to give the infection back to me"and"I wanted to let my sex partner know that he/she had given me the infection". There was a trend toward increased notification if the girls were 18 years of age or older (P = 0.07) or had only one lifetime sex partner (P = 0.08). Of the 41 subjects who notified their partners, 22 (54%) reported that the partners were treated; 16 did not know, and three knew that partners were not treated. CONCLUSIONS: The majority of inner-city girls in this study notified their partners about chlamydia infection. Self-protection from re-infection was an important reason given for notification and suggests that girls in committed ongoing relationships might be more likely to notify partners. PMID- 15749583 TI - Sexual function in women treated with dilators for vaginal agenesis. AB - OBJECTIVE: To discover the sexual satisfaction of young women treated with vaginal dilators for vaginal agenesis. DESIGN: Anonymous questionnaire study. MAIN OUTCOME MEASURES: Comparison of sexual desire arousal lubrication, orgasm satisfaction and pain with a normal population. RESULTS: There was no significant difference between the study population and controls for the domains of sexual desire, sexual arousal, and satisfaction with a sexual relationship. There was, however, a significant difference for vaginal lubrication and orgasm where the Rokitansky patients scored lower. 22.3% of patients reported some degree of dyspareunia following vaginal penetration. However, this did not affect their enjoyment of the sexual act. CONCLUSION: The use of graduated vaginal dilators for patients with Mullerian agenesis is highly successful in creating a neovagina. Although the lack of adequate lubrication, pain and difficulty in reaching orgasm is significantly higher in this group, the patients subsequently enjoy sexual satisfaction that is comparable to a normal population. PMID- 15749584 TI - What's your diagnosis? Well-demarcated vulvar erythema in two girls. PMID- 15749585 TI - Secondary amenorrhea: an unusual twist. PMID- 15749586 TI - Vulvar intraepithelial neoplasia III in a teenager. PMID- 15749587 TI - The definition of abstinence. AB - A teen's definition of sexual activity most often does not include oral or anal sex. Abstinence only programs vary widely as to how they define sexual behavior and may be contributing to misinformation about STD transmission. Unknown is the extent to which declining teen pregnancy rates are due to non-coital activities replacing vaginal intercourse. PMID- 15749588 TI - Early versus late intervention of congenital adrenal hyperplasia. PMID- 15749590 TI - Postoperative changes in body composition after gastrectomy. AB - Nutritional status is one of the most important clinical determinants of outcome after gastrectomy. The aim of this study was to compare changes in the body composition of patients undergoing laparoscopy-assisted gastrectomy (LAG), distal gastrectomy (DG), or total gastrectomy (TG). Total body protein and fat mass were measured by performing a multifrequency bioelectrical impedance analysis using an inBody II machine (Biospace, Tokyo, Japan) in 108 patients (72 men, 36 women) who had undergone LAG (n=24), DG (n=39), or TG (n=45). Changes between the preoperative data and results obtained on postoperative day 14 and 6 months after surgery were then evaluated. The mean preoperative body weight of the subjects was 57.6+/-10.7 kg, the mean body mass index was 22.5+/-3.4 kg/m(2), and the mean fat % was 24%+/-7%. In the immediate postoperative period (14 days), the body weight loss in the LAG group was significantly lower than in the DG and TG groups (2.5+/-0.9 kg vs. 3.5+/-1.8 kg and 4.0+/-1.9 kg, respectively; P < 0.0001). The body composition studies demonstrated a loss of total body protein rather than fat mass. Six months after surgery, body weight was not significantly different from preoperative values in the LAG and DG groups (-1.2+/-3.8 kg and -1.8+/-4.7 kg, respectively), but had decreased by 8.9+/-4.9 kg in the TG group (P=0.0003). A body composition analysis revealed a loss of fat mass in the DG and TG groups. The patients who underwent gastrectomy lost body protein mass during the early postoperative period. The type and extent of surgery has an effect on long-term body mass and composition. Bioelectric impedance analysis can be used to assess body composition and may be useful for nutritional assessment in patients who have undergone gastrectomy. PMID- 15749591 TI - Systematic follow-up after curative surgery for colorectal cancer in Norway: a population-based audit of effectiveness, costs, and compliance. AB - In this study, we analyzed the Norwegian guidelines for systematic follow-up after curative colorectal cancer surgery in a large single institution. Three hundred fourteen consecutive unselected patients undergoing curative surgery for colorectal cancer between 1996 and 1999 were studied with regard to asymptomatic curable recurrence, compliance with the program, and cost. Follow-up included carcinoembryonic antigen (CEA) interval measurements, colonoscopy, ultrasonography of the liver, and radiography of the chest. In 194 (62%) of the patients, follow-up was conducted according to the Norwegian guidelines. Twenty one patients (11%) were operated on for curable recurrence, and 18 patients (9%) were disease free after curative surgery for recurrence at evaluation. Four metachronous tumors (2%) were found. CEA interval measurement had to be made most frequently (534 tests needed) to detect one asymptomatic curable recurrence. Follow-up program did not influence cancer-specific survival. Overall compliance with the surveillance program was 66%, being lowest for colonoscopy (55%) and highest for ultrasonography of the liver (85%). The total program cost was 228,117 euro (US 280,994 dollars), translating into 20,530 euro (US 25,289 dollars) for one surviving patient after surgery for recurrence. The total diagnosis yield with regard to disease-free survival after surgery for recurrence was 9%. Compliance was moderate. Whether the continuing implementation of such program and cost are justified should be debated. PMID- 15749592 TI - The mechanism of microsatellite instability is different in synchronous and metachronous colorectal cancer. AB - MLH1 promoter hypermethylation has been described as the primary mechanism for high-frequency microsatellite instability (MSI-H) in sporadic colorectal cancers (CRCs). The underlying molecular mechanism for microsatellite instability (MSI) in synchronous and metachronous CRCs is not well described. A total of 33 metachronous CRC patients and 77 synchronous CRC patients were identified from 2884 consecutive patients undergoing cancer surgery in an academic center. Evaluable tumors were tested for MSI, immunohistochemistry for MLH1 and MSH2 protein expression, and hypermethylation of the MLH1 promoter. MSI-H tumors were found in 12 (36%) metachronous CRC patients and 29 (38%) synchronous CRC patients. MSI-H metachronous CRC patients were younger at index cancer diagnosis (64 vs. 76 years, P=0.01) and more often were diagnosed before 50 years of age (4 of 12 vs. 0 of 29, P=0.005). Loss of MLH1 expression associated with promoter hypermethylation was common in all patients, although more common in MSI-H synchronous patients (50% metachronous vs. 83% synchronous, P=0.03). Overall, MLH1 promoter hypermethylation was seen in 7 of 17 (41%) metachronous and 44 of 54 (81%) synchronous MSI-H CRCs tested (P=0.004). Although MSI occurred with equal frequency among patients with synchronous and metachronous CRCs, the underlying mechanism for MSI was different. Observed differences in MLH1 promoter hypermethylation and patient characteristics suggest most MSI-H synchronous CRCs in our population were sporadic in origin. In contrast, more MSI-H metachronous CRCs were associated with patient and tumor characteristics suggestive of underlying hereditary nonpolyposis CRC. PMID- 15749593 TI - Thymidylate synthase gene polymorphism as a prognostic factor for colon cancer. AB - The human thymidylate synthase (TS) gene promoter is polymorphic, having either double or triple tandem repeats of a 28-base-pair (bp) sequence. Here, we determined the significance of this polymorphism in predicting the clinical outcomes for patients with colon cancer. We reviewed 121 consecutive patients with stage II or III colon cancer who underwent a curative resection. After DNA extraction from paraffin-embedded tissues, the promoter region of the TS gene was amplified by polymerase chain reaction. In addition to the conventional prognostic factors, patient survivals were compared with regard to the pattern of TS polymorphism. Sixty-eight subjects were homozygotes for the triple-repeat variant (250 bp, group A), and 53 subjects (group B) were either homozygotes for the double-repeat variant (220 bp) or heterozygotes (220 and 250 bp). We found a significant difference between groups A and B in survival (53% versus 80%, P=0.0481). The difference was particularly significant in the patients with stage III disease (41% versus 77%, P=0.0414). Tumor stage and the TS polymorphism were identified as significant prognostic factors by multivariate analysis. We found the TS polymorphism to be a significant and independent prognostic factor for colon cancer. PMID- 15749594 TI - Embolization as first-line therapy for diverticulosis-related massive lower gastrointestinal bleeding: evidence from a meta-analysis. AB - The study goal was to determine which etiologies of lower gastrointestinal bleeding (LGIB) may best be treated with superselective embolization. A meta analysis was undertaken of 25 identified publications reporting the use of embolization and an unpublished series of 12 consecutive patients with LGIB from the authors' institution. Six published series and the authors' series met selection criteria for further analysis. Multiple regression analysis demonstrated no significant difference in pooled outcomes when varying the included study, age, or embolization method on the outcome of rebleeding. The pooled odds ratio for arteriovenous dysplastic lesions and other diseases was 3.53 compared with rebleeding after localization and embolization for diverticular disease (95% confidence interval odds ratio, 1.33, 9.41; P < 0.01). Embolization for diverticular bleeding was successful in 85% of patients. In contrast, rebleeding after embolization for nondiverticular bleeding occurred in greater than 40% of patients and over a more protracted period. Embolization for LGIB is most effective for the treatment of diverticular bleeding. Caution should be used when applying embolization therapy for nondiverticular causes due to the considerably higher associated failure rate. An inpatient observation period of 2 days is suggested following embolization for diverticular bleeding. PMID- 15749595 TI - Left trisegmentectomy with reconstruction of segment 6 hepatic venous outflow using cryopreserved vein graft. AB - Liver resections that require hepatic vein reconstruction rarely occur. Options regarding venous reconstruction include primary end-to-end reconstruction, reimplantation into the vena cava, or the use of a variety of autologous or synthetic grafts. Cryopreserved vein grafts have recently become available for use. We describe a left trisegmentectomy with bile duct resection/reconstruction during which the segment 6 hepatic vein was reconstructed into the inferior vena cava using a cryopreserved vein graft. PMID- 15749596 TI - Isolated involvement of the gallbladder by Crohn's disease manifesting as acute cholecystitis. AB - A 79-year-old female presented with acute cholecystitis. Having recovered on conservative management, elective laparoscopic cholecystectomy was performed. The diagnosis of Crohn's disease of the gallbladder was a histopathological surprise. Further detailed investigation indicated this to be the isolated involvement of gallbladder. PMID- 15749597 TI - Perioperative outcome of laparoscopic left lateral liver resection is improved by using staple line reinforcement technique: a case report. AB - Current laparoscopic stapling technology still cannot prevent bile leakage (<55%) along the resection margin. A new staple line reinforcement technique was used in the present case. This technique incorporates an absorbable polymer membrane into the stapler system, such that it buttresses the transected solid organ. The objectives of this novel procedure are to decrease hemorrhage at the staple line and to prevent bile duct leakage after liver resection. A 47-year-old man followed for status post biliopancreatic diversion with duodenal switch presented with epigastric pain. On imaging, he was found to have a lesion in segment 2-3 of the left lobe of the liver, which measured at least 3 cm in diameter. He was admitted to the hospital to undergo a laparoscopic left lateral liver resection. This procedure involved laparoscopic ultrasonography of the liver and transection of the left liver lobe with endoscopic linear staplers. The staple height of 3.5, 60 mm long, reinforced with an absorbable polymer membrane was used for liver transection to catch the portal branches. This required multiple firings in the liver parenchyma and additional division of some tissue using the Harmonic scalpel. The larger branch in the middle of segment 2-3 and the left hepatic vein were both transected with the novel staple line reinforcement technique. Bleeding or any bile leakage in this area could not be visualized. No drains were left. The patient's postoperative course was uncomplicated, and he was discharged on postoperative day 3. Pathology results showed a cavernous hemangioma of 4.5 cm in diameter. Staple line reinforcement with the absorbable polymer membrane has the potential to decrease staple line hemorrhage and bile leakage. PMID- 15749598 TI - Portal venous reconstruction in a living liver donor with an anomalous hepatic arterial and portal venous anatomy. AB - Hepatic arterial and portal venous anomalies in living liver donors are not uncommon. Modified surgical techniques may be required in such circumstances, although the safety of the living donor must always be given top priority. We describe here a successful portal venous reconstruction in a living donor with an anomalous hepatic arterial and portal venous anatomy in which the right anterior and posterior hepatic arteries encircled the main portal vein. Although such an anomaly of hepatic vessels was not frequently encountered, we should be able to alter the surgical strategy to deal with it. This case illustrates the importance of preoperative hepatic artery and portal venous evaluation in all living donors to identify the feasibility of modifying vessel anastomoses in living donors, as well as recipients, before living donor liver transplantation. PMID- 15749599 TI - Bile duct replacement using an autologous femoral vein graft: an experimental study. Preliminary results. AB - The repair of common bile duct injuries is a complex procedure with a significant rate of postoperative morbidity and mortality. The aim of this study was to demonstrate the usefulness of the autologous vein graft in replacement of the bile duct. Twelve male Sprague-Dawley rats weighing 350+/-550 g were used in the study and were divided at random into two groups: the control group (60) and the experimental group in which a 3-mm segment of the bile duct was resected and the biliary tract was replaced by a segment of vein aided by stent (G1). Both groups were subdivided into pairs of rats to study at 30, 60, and 120 days. All of the animals underwent radioisotope cholangiography, a repeat laparotomy, and blood tests for further pathologic study. The clinical evaluation and biochemical nuclear medicine and pathologic studies showed no evidence of cholestasis. The histologic study of the graft showed replacement of the endothelium by biliary appearing epithelium. The use of an autologous vein graft with a supporting stent proves to be a feasible and alternative procedure for bile duct reconstruction. Further experimental studies should be carried out to validate these findings so they can be implemented in clinical cases. PMID- 15749600 TI - Analysis of preoperative prognostic factors for long-term survival after hepatic resection of liver metastasis of colorectal carcinoma. AB - Hepatic resection is the most effective therapy for liver metastasis of colorectal carcinoma. To clarify indications for this therapy, the clinicopathologic and follow-up data of 103 consecutive patients who underwent hepatic resection for metastases of colorectal carcinoma were analyzed. Factors influencing overall survival rate were investigated by multivariate analysis. Thereafter, patients who underwent resection were stratified according to the number of independent risk factors present, and their outcomes were compared with those of 14 nonresection patients with fewer than six liver tumors and without extrahepatic metastasis. The overall survival rate of the 103 resection patients was 43.1%. The clinicopathologic factors shown to affect on long-term survival after hepatic resection were the interval between colorectal and hepatic surgery (<12 months), preoperative carcinoembryonic antigen level (> or =10 ng/ml), and number of hepatic metastases (four or more). The 5-year overall survival rates were 75.0% with no risk factors (n=16), 53.6% with one risk factor (n=46), 23.0% with two risk factors (n=36), and 0% with three risk factors (n=5). Survival rates did not differ between resection patients with three risk factors and nonresection patients. Therefore, hepatic resection may be appropriate for patients with fewer than three risk factors. PMID- 15749601 TI - Laparoscopic surgery in patients with sporadic and multiple insulinomas associated with multiple endocrine neoplasia type 1. AB - There have recently been reports of a limited number of laparoscopic procedures in patients with clinically manifest hyperinsulinism. However, the precise role of laparoscopy remains unknown. Between January 1998 and September 2003, 11 consecutive patients (10 women and 1 man; mean age, 40 years; age range, 22-66 years) with sporadic insulinoma and two female patients (25 and 40 years old) with multiple insulinomas associated with multiple endocrine neoplasia type 1 (MEN-1) were operated on using the laparoscopic approach. Endoscopic ultrasonography was used to localize the tumor preoperatively in 90% of patients with sporadic insulinoma. In patients with MEN-1, computed tomography and octreoscan-(111)In demonstrated multiple tumors. Laparoscopic ultrasonography (LapUS) was performed in all patients for operative decision-making. Of 11 patients with sporadic insulinoma, laparoscopic enucleation (LapEn) was planned in 8 patients, but in 1 patient, the use of LapUS missed the tumor and the patient was converted to open surgery. Mean operating time after LapEn (seven patients) was 180 minutes, and the mean blood loss was 200 ml. The mean hospital stay was 5 days. In three of the 11 patients, laparoscopic spleen-preserving distal pancreatectomy (LapSPDP) was performed; the mean operative time was 240 minutes, and the mean blood loss was 360 ml. Postoperative complications occurred in three of seven patients after LapEn (three pancreatic fistulas managed conservatively, and one case of bleeding requiring reoperation). LapSPDP was performed in both patients with MEN-1; in one patient with splenic vessel preservation (SVP), the operating time was 210 minutes and blood loss was 650 ml, with a hospital stay of 6 days. In another patient without SVP, the operating time was 150 minutes and blood loss was 300 ml. The latter patient developed a 4 cm splenic infarct managed conservatively, and the hospital stay was 14 days. LapEn and LapSPDP are feasible and safe and achieved cure in patients with sporadic insulinoma and multiple insulinomas associated with MEN-1. However, the risk of pancreatic leakage after LapEn remains high, and LapSPDP without SVP may be associated with splenic infarct. PMID- 15749602 TI - Duodenal cancer arising from the remaining duodenum after pylorus-preserving pancreatoduodenectomy for ampullary cancer in familial adenomatous polyposis. AB - We herein report a rare occurrence of duodenal cancer arising from the remaining duodenum after pylorus-preserving pancreatoduodenectomy for ampullary cancer in familial adenomatous polyposis (FAP). In this patient, proctocolectomy and ileoanal anastomosis for FAP had been performed 11 years earlier. During the current admission, the patient was diagnosed with adenocarcinoma in the Vater's ampulla using imaging and pathological examinations. In addition, a pylorus preserving pancreatoduodenectomy was performed. The tumor was a well differentiated tubular adenocarcinoma and no other polyps were identified in the duodenum by pathological examination. However, 1 year after surgery, a polypoid lesion measuring 15 x 15 mm was indicated in the remaining duodenum by endoscopic surveillance. This lesion was completely resected by endoscopic mucosal resection and the resected specimen revealed well-differentiated tubular adenocarcinoma in an adenomatous lesion. This report suggests that resection of the total duodenum is essential for duodenal neoplasms in FAP to prevent a recurrence in the remaining duodenum. PMID- 15749603 TI - Transarterial embolization for postoperative hemorrhage after abdominal surgery. AB - The study goal was to evaluate the efficacy, safety, and clinical outcome of transarterial embolization for postoperative hemorrhage after abdominal surgery. Thirty-three patients were referred for angiography because of gastrointestinal or intra-abdominal bleeding after abdominal surgery. Urgent angiography and transarterial embolization was performed in all 33 patients. The clinical and angiographic features were retrospectively reviewed. Angiography revealed a discrete bleeding focus in 26 (79%) of 33 patients. Transarterial embolization was technically successful in 24 (92%) of 26 patients with a discrete bleeding focus. Rebleeding occurred in four (17%) of 24 patients. They were successfully managed with repeat embolization. There was no procedure-related complication during follow-up period. Angiography has a high detection rate of bleeding site in patients with postoperative hemorrhage after abdominal surgery. Transarterial embolization is considered to be an effective and safe means in the management of postoperative hemorrhage. PMID- 15749604 TI - Improved outcomes for benign disease with limited pancreatic head resection. AB - We sought to determine whether duodenum-preserving pancreatic head resections (DPPHRs) offer improved outcomes for benign disease of the proximal pancreas. A single-cohort study was performed of 86 consecutive patients who underwent DPPHR, extended lateral pancreaticojejunostomy with excavation of the pancreatic head (ELPJ), standard or pylorus-sparing Whipple procedure (WHIP), or distal pancreatectomy (DPR). Aspects of cost, complications (mortality and morbidity), and outcomes were assessed during a follow-up period of 6-63 months (mean, 3 years). Twelve DPPHR and 12 ELPJ procedures were performed for benign lesions or chronic pancreatitis (CP), as were 7 of 30 WHIP procedures and 12 of 16 DPRs. Operative time was significantly less than that for WHIP in ELPJ and DPR procedures. Major complications occurred in 40% of WHIPs and 25% of DPPHRs but only 16% of ELPJs (P < 0.05). Thirty-day mortality was 2 of 30 for WHIP but 0 for all other procedures. Pancreatic or biliary leak occurred in 3 of 30 WHIPs, 3 of 12 DPPHRs, 1 of 16 DPRs, and 0 of 12 ELPJs. New diabetes occurred in 25% of patients who underwent WHIP but only 8% of both DPPHR and ELPJ patients. Full functional recovery was similar for CP patients in both DPPHR and ELPJ. DPPHR and ELPJ are effective surgical approaches to the treatment of benign tumors and CP and are safer than WHIP with lower morbidity and mortality risks. The incidence of new diabetes is less with both ELPJ and DPPHR. PMID- 15749605 TI - Focal hyperthermia produces progressive tumor necrosis independent of the initial thermal effects. AB - Focal hyperthermia, produced using laser, radio frequency, and microwave, is used to treat liver tumors. The exact mechanisms of tissue destruction using focal hyperthermia are, however, unknown. Clinical and experimental studies suggest a progression of injury after cessation of the initial heat stimulus. This study investigates the mechanisms and time sequence of progressive tissue necrosis induced using focal hyperthermia in a murine model of colorectal liver metastases. Focal hyperthermia produced using a neodymium-yttrium aluminum garnet (Nd-YAG) laser source was applied to the normal liver and colorectal cancer liver metastases in inbred male CBA strain mice. The extent of direct lethal thermal injury was assessed histochemically using vital stain for nicotinamide adenine dinucleotide (NADH) diaphorase immediately after laser application. Tissue injury at subsequent time points was assessed using both NADH diaphorase staining and routine histology to determine the temporal relationship between tissue necrosis and time. Thermal injury occurring immediately after the application of 100 joules of energy was greater in the tumor tissue than in the normal liver (mean [standard error of the mean (SEM)]), measuring 23.5 (3.4) and 16.3 (2.6) mm(3), respectively (P=0.046), despite similar tissue temperature profiles. There was a significant increase in tissue necrosis after initial injury that was greater in the normal liver than in the tumor tissue. In the normal liver, the peak volume of necrosis was 137.4 (9.8) mm(3) and occurred at 3 days, whereas in the tumor tissue the peak was 49.0 (3.5) mm(3) at 4.5 days (P < 0.001). Focal hyperthermia produces tissue necrosis that occurs in two phases. The first phase is caused by the direct lethal thermal injury followed by a second phase involving a progression of necrosis beyond the initial thermal effects. The normal liver and the tumor tissue responded differently to focal hyperthermia. In the tumor tissue, the direct injury is more pronounced, whereas the progression of injury is more rapid and extensive in the normal liver. PMID- 15749606 TI - Analysis of prognostic and immunohistochemical factors in gastrointestinal stromal tumors with malignant potential. AB - The aim of this study was to analyze 37 patients with malignant primary gastrointestinal stromal tumors and to compare the findings and their therapeutic implications with those previously reported. The medical records of 37 patients who were diagnosed and operated on between January 1996 and December 2002 were retrospectively reviewed. The patients' age, tumor size, type of surgery, histologic type, mitotic counts, presence of necrosis, Ki-67 proliferative index, National Institutes of Health 2001 consensus classification, immunohistochemical staining, and recurrence were examined to analyze factors affecting survival. Overall actuarial survival for all patients was 46%. When analyzed by type of resection, the complete resection group (R0 resection) had a mean overall survival of 48.2 +/- 6.18 months compared with the patients with incomplete resection (R1-R2) who survived a mean of 10.8 +/- 3.2 months (P=0.00). Univariate analysis showed development of recurrence (P=0.00), tumor size of 8 cm or greater (P=0.05), Ki-67 proliferative index greater than 0.82 (P=0.0448), desmin staining (P=0.0076), age younger than 49 years (P=0.0009), and incomplete resection (P=0.00) to be significantly correlated with a poor survival. In multivariate analysis, desmin staining (P=0.031), tumor size (P=0.033), age (P=0.01), recurrence (P=0.038), and R0 resection (P=0.02) were significant independent prognostic factors. We recommend that more careful preoperative and more frequent postoperative follow-up examinations be performed for patients with large tumors, age of younger than 49 years, and Ki-67 proliferative index greater than 0.82. PMID- 15749607 TI - Perforation through small bowel malignant tumors. AB - Data on 19 patients (6 women and 13 men) with malignancy perforation through small bowel tissue were retrospectively reviewed. The median patient age was 57 years (range, 41-81 years). The histopathology included lymphoma (seven patients), leiomyosarcoma (two patients), gastrointestinal stromal tumor (one patient), adenocarcinoma (one patient), metastatic carcinomas with unknown primary tumor (four patients), metastatic adenocarcinoma from the lung (one patient), and metastatic carcinomas from the hypopharynx (one patient), cervix (one patient), and lung (one patient). Resection of a segment of perforated bowel with primary anastomosis was performed in 16 patients, wedge resection of perforated lesion with plication in two patients, and loop ileostomy in one patient. Postoperative deaths occurred in 10 (52.6%) patients, owing to sepsis and organ functional failure. Seven patients died from the primary malignancy at a median follow-up of 6.5 months (range, 5 months to 1 year 9 months) after surgery. Moreover, two patients with small bowel lymphoma were alive with disease at 4 years 8 months and 7 years 1 month after surgery. In conclusion, perforation through small bowel malignant tumors had a high postoperative mortality rate. High index of suspicion of the disease with early surgical treatment may improve treatment outcomes. PMID- 15749608 TI - Pancreatic necrosectomy: definitions and technique. AB - Pancreatic necrosis implies a permanent condition in which a portion of the pancreas loses its blood supply. This condition is irreversible, yet many cases of "necrosis" will, after recovery, culminate in a patient with a normal pancreas by computed tomography or endoscopic retrograde cholangiopancreatography. The problem is in our definitions. An understanding of this disease through its related definitions is required before judgment deems "necrosectomy to be appropriate." These definitions are of pancreatic ductal disruption, peripancreatic fluid collections, pseudocyst, pancreatic abscess, and pancreatic necrosis. The technique of necrosectomy removes mature "necrosum" and is described in this article. Once necrosectomy is completed, the surgeon still depends on the continued support of interventional radiology through regular exchange of large-bore pancreatic drains. In our institution, many of these drain sites are placed at some time before necrosectomy. Once the team method has been implemented, the following improved outcomes will result--lowered need for necrosectomy and single digit mortality. PMID- 15749611 TI - Proposal of a modified, treatment-oriented classification of odontoid fractures. AB - BACKGROUND CONTEXT: The classification scheme of odontoid fractures described by Anderson and D'Alonzo is the one most commonly used. However, uncertainty exists in the distinction between Type II and "shallow" Type III fractures. Moreover, fractures at the base of the odontoid (Anderson and D'Alonzo Type II) include a spectrum of injury patterns. PURPOSE: To modify the Anderson and D'Alonzo classification of odontoid fractures based on current clinical treatment options. STUDY DESIGN: Proposal of a modified classification system for odontoid fractures. METHODS: A more precise distinction between Type II and III fractures based on the presence/absence of C1-C2 facet involvement is proposed. A modified classification of Type II fractures based on fracture line obliquity, displacement and comminution is then proposed, because these are factors deemed to influence management. To evaluate the reproducibility of this classification, 52 odontoid fractures were reviewed and classified by four attending spine surgeons and three spine fellows. RESULTS: There was substantial agreement (at least five of seven respondents) in 70% of cases. The overall kappa value for the modified classification system was 0.48, indicating moderate agreement, and there were no differences in kappa values between attending spine surgeons and fellows. CONCLUSIONS: The reproducibility of this system was demonstrated by the moderate agreement observed when applied to odontoid fractures at our institution. The proposed utility of this system is its ability to guide clinical decision making in the treatment of odontoid fractures. Prospective application of this modified classification system and suggested treatment options is now required. PMID- 15749612 TI - Effect of head rotation in whiplash-type rear impacts. AB - BACKGROUND CONTEXT: Knowledge is increasing about the electromyographic and kinematic response of the neck muscles to rear impact, and also recent information is available on the effect of a rear impact offset to the left (posterolateral). The effect of head rotation, however, at the time of rear impact is not known. PURPOSE: The purpose of this study was to examine the effects of head rotation to the left and right on the cervical muscle response to increasing low-velocity posterolateral impacts. STUDY DESIGN/SETTING: Twenty healthy volunteers were subjected to rear impacts of 4.7, 8.3, 10.9 and 13.7 m/s2 acceleration, offset by 45 degrees to the subject's left, with head rotation to right and left. METHODS: Bilateral electromyograms of the sternocleidomastoids, trapezii and splenii capitis were recorded. Triaxial accelerometers recorded the acceleration of the sled, torso at the shoulder level, and head of the participant. RESULTS: With the head rotated to the right, at an acceleration of 13.7 m/s2, the left sternocleidomastoid generated 59% and the right sternocleidomastoid 20% of their maximal voluntary contraction (MVC) electromyogram (EMG). Under these conditions, the remaining muscles (both splenii capitis and trapezius) generated 25% or less of their MVC. With the head rotated to the left, at an acceleration of 13.7 m/s2, the right sternocleidomastoid generated 65% and the left sternocleidomastoid only 11% of the MVC EMG. Under these conditions, again the remaining muscles had low EMG activity (27% or less) with the exception of the left trapezius which generated 47% of its MVC. Electromyographic variables were significantly affected by the levels of acceleration (p<.01). The time to onset and time to peak EMG for all muscles progressively decreased with increasing levels of acceleration, for both head rotation conditions. The kinetic variables and the electromyographic variables regressed significantly on the acceleration (p<.01). CONCLUSIONS: Direction of impact is a factor in determining the muscle response to whiplash, but head rotation at the time of impact is also important in this regard. More specifically, when a rear impact is left posterolateral, it results in increased EMG generation mainly in the contralateral sternocleidomastoid, as expected, but head rotation at the same time in this type of impact reduces the EMG response of the cervical muscles. Muscle injury seems less likely under these conditions in low-velocity impacts. PMID- 15749613 TI - Surgical treatment of dysphagia after anterior cervical interbody fusion. AB - BACKGROUND CONTEXT: Dysphagia is a frequent complication after anterior cervical interbody fusion (ACIF). Although dysphagia usually improves over 6 months, it remains a significant and persistent problem for some patients. The etiology is poorly understood but has been reported to be associated with vocal cord paralysis, dislodgement of instrumentation and unidentified causes, such as hematoma, adhesion formation and denervation of the pharyngeal plexus. A surgical treatment of dysphagia after ACIF has not been reported. PURPOSE: We report the surgical treatment of persistent dysphagia occurring after ACIF with instrumentation. STUDY DESIGN/SETTING: A retrospective review of cervical discectomy and interbody fusion patients identified a subset of patients with symptomatic dysphagia who chose surgical treatment of the dysphagia. The hypothesis is that removal of the anterior cervical plate will release mechanical adhesions of the esophagus to the anterior spine around the plate. Outcome was graded by examination and a final telephonic interview with a dysphagia questionnaire. METHODS: Thirty-one patients who elected surgical treatment for persistent dysphagia were assessed at clinic visits after surgery at 3, 6 and 12 months for symptomatic dysphagia, and with a final telephonic questionnaire. The average time from initial surgery to time of surgical treatment for dysphagia was 18 months. Final follow-up was an average 11 months (range, 6 to 25 months) with a dysphagia questionnaire using the Bazaz-Yoo dysphagia score. Thirty-one patients responded to a phone questionnaire with the Bazaz-Yoo dysphagia score. RESULTS: The primary operative finding was extensive adhesions attaching the esophagus to the prevertebral fascia and anterior cervical spine around the periphery of the cervical plate. Seventeen patients (55%) were significantly improved to no dysphagia of solids and liquids (p < or = .0001). Ten patients (32%) reported mild dysphagia occasionally with specific foods. Three patients had persistent moderate occasional dysphagia with solid food. Two patients had persistent severe dysphagia of solids and liquids. Previous cervical surgery, particularly with pre-existing dysphagia, and unexpectedly extreme amounts of adhesions at surgery were contributing factors to the cases with persistent severe dysphagia. CONCLUSIONS: Surgical treatment of dysphagia after ACIF has not been reported. Removal of the cervical instrumentation in patients will improve the dysphagia. This improvement with surgical management, as compared with the dissatisfaction before surgical treatment, documents that this surgical treatment is a reasonable option. PMID- 15749609 TI - Current management of acute pancreatitis. PMID- 15749614 TI - Effects of prolonged sitting on the passive flexion stiffness of the in vivo lumbar spine. AB - BACKGROUND CONTEXT: Prolonged sitting may alter the passive stiffness of the lumbar spine. Consequently, performing full lumbar flexion movements after extended periods of sitting may increase the risk of low back injury. PURPOSE: The purpose was to quantify time-varying changes in the passive flexion stiffness of the lumbar spine with exposure to prolonged sitting and to link these changes to lumbar postures and trunk extensor muscle activation while sitting. A secondary objective was to determine whether men and women responded differently to prolonged sitting. STUDY DESIGN: Passive lumbar flexion moment-angle curves were generated before, during and after 2 hours of sitting. Lumbar flexion/extension postures and extensor muscle activation levels were measured while sitting. SAMPLE: Twelve (6 men, 6 women) university students with no recent low back pain were studied. OUTCOME MEASURES: Quantified changes in the shapes of the passive flexion moment-angle curves (slopes, breakpoints and maximum lumbar flexion angles) were the outcome measures. While sitting, average lumbar flexion/extension angles, the distribution of lumbar flexion/extension postures, average electromyogram (EMG) amplitude, the number and average length of EMG gaps, and trunk extensor muscle rest levels were measured. METHODS: Participants performed deskwork for 2 hours while sitting on the seat pan of an office chair. Moment-angle relationships for the lumbar spine were derived by pulling participants through their full voluntary range of lumbar flexion on a customized frictionless table. RESULTS: Lumbar spine stiffness increased in men after only 1 hour of sitting, whereas the responses of women were variable over the 2-hour trial. Men appeared to compensate for this increase in stiffness by assuming less lumbar flexion in the second hour of sitting. CONCLUSIONS: Changes in the passive flexion stiffness of the lumbar spine may increase the risk of low back injury after prolonged sitting and may contribute to low back pain in sitting. PMID- 15749615 TI - Three- to six-year follow-up of stand-alone BAK cages implanted by a single surgeon. AB - BACKGROUND CONTEXT: Multiple studies involving the outcomes of anterior interbody cages have been published, but the majority were by authors who designed the cage. No outcome studies with Bagby and Kuslich (BAK) cages implanted by a single surgeon have either 3 years of follow-up or at least 25 patients. PURPOSE: To determine the 3- to 6-year clinical outcomes, including fusion rate, revision rate, complications and functional status of patients who underwent placement of anterior, stand-alone BAK cages by a single surgeon. STUDY DESIGN: This is a retrospective cohort study of patients who underwent anterior, stand-alone BAK cage placement by a single surgeon with a minimum of 3 years of follow-up. PATIENT SAMPLE: A total of 46 consecutive patients who underwent placement of anterior BAK cages from 1997 to 1999 were the study group. OUTCOME MEASURES: Complications, fusion rate, revision rate, Prolo scores, Oswestry scores, pain scores, patient satisfaction. METHODS: A retrospective review of charts of patients undergoing anterior stand-alone placement of BAK cages over the study period were reviewed. Demographic data were collected, and postoperative radiographs and clinic notes were reviewed. Patients were then contacted to complete a survey to determine Prolo and Oswestry scores, workers' compensation status and general satisfaction with the procedure. Patients were also asked to undergo repeat radiographs of their lumbar spine to determine if the operative levels had fused. RESULTS: Follow-up was available on 33 of 46 patients (72%). At least 10 patients (22%) required revision surgery. Ten patients (22%) had 14 total complications not requiring revision surgery. Seventy percent of patients had a fair or poor outcome as assessed by the Prolo rating system, and 58% of patients had at least "severe disability" according to the Oswestry outcome scale. Fifty percent of patients were satisfied with their surgery. CONCLUSIONS: This single-surgeon series of stand-alone BAK cages demonstrates significantly worse clinical outcomes than has been previously reported. The use of stand-alone BAK cages for degenerative disc disease should be reconsidered given the large number of patients with unacceptable outcomes. PMID- 15749617 TI - Posterior lumbar interbody fusion for degenerative spondylolisthesis: restoration of sagittal balance using insert-and-rotate interbody spacers. AB - BACKGROUND CONTEXT: Although satisfactory clinical outcomes have been reported for degenerative spondylolisthesis using a variety of surgical techniques, its optimal management remains controversial. Laboratory and clinical evidence is emerging that if fusion surgery is undertaken, improved short- and longer-term outcomes may be achieved by correcting any sagittal deformity present. The insert and-rotate posterior lumbar interbody fusion (PLIF) technique, first described by Jaslow in 1946, may enable surgeons to safely and effectively correct sagittal balance through a single posterior approach. PURPOSE: To examine the clinical outcomes and determine whether the focal sagittal imbalance associated with a degenerative lumbar spondylolisthesis can be safely and effectively corrected using a posterior distraction/reduction technique and insert-and-rotate interbody fusion spacers. STUDY DESIGN/SETTING: A prospective, single-cohort, observational study of the clinical outcomes and retrospective radiological review, in a series of 34 patients with degenerative spondylolisthesis, who underwent surgery between September 2000 and October 2002. PATIENT SAMPLE: Mean age of 65.1 years (range, 35 to 82 years). Thirty-two of the 34 patients underwent surgery principally for the relief of radicular leg pain. The principal indication for the fusion was the prevention of anticipated postdecompression instability in 68% of the patients. Mean preoperative slip was 20.0% (range, 12% to 33%). Mean preoperative focal lordosis was 13.2 degrees. OUTCOME MEASURES: Patients were administered pre- and postoperative clinical outcome surveys recording Visual Analogue Pain Score (VAS), Low Back Outcome Score (LBOS), Short Form (SF)-12 and patient satisfaction questionnaires. Pre- and postoperative measurements of the percentage slip and lumbar lordosis of the involved segments were available on 17 patients. Statistical analysis was done using a two tailed, paired t test. SURGICAL METHODS: Decompressive laminectomy was followed by reduction of the spondylolisthesis using intervertebral disc space distraction and pedicle screw instrumentation. The vertebral bodies were supported with bilateral intervertebral lordotic spacers, inserted on their sides and rotated 90 degrees before placing bone graft to either side of them, within the disc space. RESULTS: Mean follow-up time was 21.2 months (range, 12-32 months). No patients were lost to follow-up. Mean preoperative measures of VAS and LBOS of 5.3+/-2.2 and 24.8+/ 15.6, respectively, improved to 2.2+/-2.1 (p<.001) and 44.8+/-18.0 (p<.001) at last follow-up. Thirty-one of 34 patients (91%) considered their outcome to be good or excellent. Mean preoperative slip reduced from 20.2% to 1.7% (92% correction, p<.001). Mean focal lordosis increased from 13.1 to 16.1 degrees (26.0% increase, p=.01). There were no device-related procedural complications. Postoperatively, three patients developed an ileus and one a possible deep wound infection, which settled on antibiotics. One patient required extension of his fusion at 12 months for adjacent segment stenosis. CONCLUSIONS: This series suggests that PLIF for degenerative spondylolisthesis using an insert-and-rotate technique can yield high levels of patient satisfaction with low levels of complications and substantial deformity correction using a posterior only approach. Longer-term outcome studies are required regarding the relevance of the restoration of spinal balance. PMID- 15749616 TI - Posterior lumbar interbody fusion for lytic spondylolisthesis: restoration of sagittal balance using insert-and-rotate interbody spacers. AB - BACKGROUND CONTEXT: The role of surgical correction of sagittal plane deformity in cases of lytic spondylolisthesis remains controversial. While some early evidence is emerging of the possible short- and long-term benefits of restoring spinal balance, many surgeons have been concerned about the associated risks. The insert-and-rotate posterior lumbar interbody fusion (PLIF) technique, first described by Jaslow in 1946, may enable surgeons to safely and effectively correct sagittal balance through a single posterior approach. PURPOSE: To determine whether the focal kyphosis and subluxation associated with a lytic lumbosacral spondylolisthesis can be safely and effectively corrected using a single-stage posterior distraction/reduction technique and insert-and-rotate interbody fusion spacers. STUDY DESIGN/SETTING: A prospective, single cohort, observational study of the clinical outcomes and retrospective radiological review, in a series of 18 consecutive patients with lytic spondylolisthesis Grades I to IV, operated between September 2000 and December 2002. PATIENT SAMPLE: Mean age of 50.2 years (range, 15.5 to 77.8 years). Principal indication for surgery was relief of radicular pain secondary to foraminal stenosis in 16 of 18 patients, and back pain was the principal symptom in 2 patients. Mean preoperative slip was 30.2% (range, 9% to 78%). Mean preoperative focal lordosis was 10.6 degrees (range, -12 to 33 degrees). OUTCOME MEASURES: Minimum 12-month follow-up was available on all patients except one, who died of unrelated causes after his 6-month visit. Patients completed Visual Analogue Pain Score (VAS), Low Back Outcome Score (LBOS), Short Form (SF)-12 and patient satisfaction questionnaires. Pre- and postoperative measurements of the percentage slip and lumbar lordosis of the involved segments were available on 13 patients. SURGICAL METHODS: Decompressive laminectomy was followed by reduction of the spondylolisthesis with the aid of intervertebral disc space spreaders and supplementary pedicle screw instrumentation. The vertebral bodies were supported with bilateral intervertebral lordotic spacers made from carbon fiber, titanium mesh or polyether-ether-ketone (Medtronic Sofamor Danek, Memphis, TN). These were inserted on their sides and rotated 90 degrees before placing bone graft to either side of them, within the disc space. RESULTS: Mean follow-up was 17.3 months. Mean preoperative measures of VAS and LBOS of 5.0+/-2.3 and 29.3+/-16.4, respectively, improved to 2.9+/-3.0 (p=.01) and 42.6+/-20.1 (p=.005) at last follow-up. Fifteen of 18 patients (83.3%) considered their outcome to be good or excellent. Mean preoperative slip reduced from 30.2% to 6.2% (p=.001). Mean focal lordosis improved from 10.6 to 18.1 degrees (p=.02). Lumbar lordosis (L1-S1) did not change, but the lordosis of the lumbar segments above the fusion reduced from 46.8 to 34.9 degrees (p=.02). There were no serious implant or procedural complications. Postoperatively, there was a delayed and temporary weakness of ankle dorsiflexion in a patient with Grade IV spondylolisthesis. CONCLUSIONS: This series suggests that PLIF using an insert-and-rotate technique can yield satisfactory clinical outcomes and substantial deformity correction using a posterior only approach and with low levels of serious or permanent complications. Longer-term clinical outcome and comparative studies are required regarding the importance of the restoration of spinal balance. PMID- 15749618 TI - Topical high-molecular-weight hyaluronan and a roofing barrier sheet equally inhibit postlaminectomy fibrosis. AB - BACKGROUND CONTEXT: The relevance of epidural fibrosis to failed back surgical outcomes remains controversial. Previous studies on the correlation between epidural fibrosis and clinical outcome after laminectomy are inconclusive, and clinical approaches applied to reduce postlaminectomy spinal canal scarring have produced mixed outcomes. PURPOSE: Improved preclinical models are required to address the fundamental question of the relationship between postlaminectomy fibrosis and chronic pain. This study is directed at establishing small animal postlaminectomy models characterized by significantly reduced scar within the spinal canal postoperatively. Such preclinical models are offered as a platform for future studies to explore the potential relationship between postlaminectomy epidural fibrosis and persistent neuropathy with its potential for altered spinal mechanisms for pain processing, so-called spinal facilitation. Such experiments could be constructed in these models for comparison of pain behavior and its underlying neurochemistry both in the presence and absence of extensive postlaminectomy epidural scar. STUDY DESIGN/SETTING: A modified rat laminectomy model was employed to assess epidural fibrosis using a quantitative biochemical collagen assessment approach along with correlative histology. This group served as the control for comparison with groups in which antifibrotic measures were employed. We compared antifibrotic efficacy of a bioabsorbable roofing barrier sheet placed over the laminectomy defect with topical high-molecular-weight hyaluronan (HMW HA) gel, each applied postoperatively to prevent proliferative epidural scarring. Routine biomechanical tensile strength testing was employed to assess wound-healing strength. METHODS: A bilateral laminectomy (L5 and L6) with associated unilateral disc injury (L5-L6) was performed in 98 male Harlan Sprague Dawley rats. The laminectomy models described incorporated a unilateral disc injury at L5-L6 because herniated disc material has been shown to contribute proinflammatory cytokines in the postoperative wound. Five groups were employed for the study: 1) normal controls without surgery; 2) a laminectomy-disc injury group without treatment; 3) a laminectomy-disc injury group treated with topical HMW HA gel; 4) a laminectomy-disc injury group treated with 0.2-mm thick bioabsorbable roofing barrier sheet in which a protected space was maintained between overlying paraspinous muscles and the dura and 5) a 0.02-mm thin barrier sheet treatment group in which the sheet was placed directly on the dura. The animals were sacrificed at 3- and 8-week postoperative intervals for analysis. The dissected specimens were studied biochemically for hydroxyproline content to estimate total collagen within the canal and on the dura between L4 and L7. Additional specimens were prepared histologically and stained with Masson-Goldner Trichrome stain to confirm presence of proliferative collagen and to describe the presence or absence of wound-healing scar adherence to the dura. The surgical incisions were studied biomechanically by uniaxial tensile testing to determine ultimate force, strain and prefailure stiffness. Statistics were performed using analysis of variance. RESULTS: Gross appearance and histology studies showed that the untreated laminectomy group demonstrated postoperative scar formation that is adherent between the wound and the dorsum of the dura mater in both 3- and 8-week groups. Proliferative scar was substantially increased grossly between the 3- and 8-week intervals. By gross observation there was adherence of the L5 spinal nerve to the underlying disc and adjacent pedicle on the disc injury side. Gross observation of treatment groups, in contrast, disclosed that both the 0.2-mm thick roofing barrier sheet and topical HMW HA gel each prevented scar attachment to the dural sleeve at both the 3- and 8-week postoperative intervals. Furthermore, both the HMW HA gel and 0.2-mm thick roofing barrier sheet treatment groups had significant reduction of total collagen content in the laminectomy specimens measured biochemically at the two time periods compared with the untreated controls. Histologically, the HMW HA gel and the 0.2-mm thick barrier sheet findings were consistent with the gross observations concerning lack of adherence between scar of the overlying wound and the dura. Notably, both the 0.2 and the 0.02-mm barrier sheets became enveloped by a fibrotic envelope consistent with a foreign body reaction. In the group in which the 0.02-mm thin sheet was placed within the canal on top of the dura, there was an increase of fibrosis around the sheet within the canal leading to a space-occupying mass within the canal. Although the 0.2-mm thick roofing barrier placed external to the canal became enveloped by scar, it appeared to attract proliferative scar away from the epidural space, leaving the dura relatively free of scarring or adherence to overlying tissues. The mechanical properties of the incisional wound increased significantly between 3 and 8 weeks. The ultimate strength, stress, strain and stiffness of the several groups were similar at each time point. CONCLUSION: These results provide two preclinical rat laminectomy models of potential usefulness for the future study of the relevance of epidural fibrosis to behaviorally defined pain states, and for the study of the potential of an altered neurochemical signature in postlaminectomy pain conditions. Such preclinical models have become standard in studies of pain behavior and its neurochemistry in preclinical sciatic nerve and spinal nerve injury models, and should be of utility in the studies of postlaminectomy fibrosis. There was progressive scar proliferation and maturation in the untreated postlaminectomy group in the postoperative interval between 3 and 8 weeks. HMW HA gel applied topically and a 0.2-mm thick bioabsorbable Macropore sheet used as a roofing barrier each significantly reduced postlaminectomy proliferative scar without affecting the integrity of incisional wound healing. However, if the 0.02-mm thin barrier sheet used in this study is placed within the canal in contact with the dura and adjacent to the pedicles, the process of reabsorption results in a fibrotic mass within the canal. The preferred barrier sheet placement for this model is clearly in a roofing position bridging over the open epidural space. It must be placed in a manner to block off the paraspinous muscle healing response and still leave a gap between the sheet and the dura. PMID- 15749619 TI - Epidural steroid therapy for back and leg pain: mechanisms of action and efficacy. AB - BACKGROUND CONTEXT: Epidural steroid injection (ESI) is one of the most common nonsurgical interventions prescribed for back and leg pain symptoms. Although the use of ESI is widespread, proof of efficacy among the broad population of low back pain patients is lacking and use is predicated to a great extent on the cost and morbidity of the perceived "next step" in many patient's care-surgery. PURPOSE: To review the relative indications and clinical features that predict success with ESI therapy, and to provide a physiological rationale to guide clinical decision-making. STUDY DESIGN/SETTING: Review of literature and clinical experience. RESULTS: Clinical studies have alternately supported and refuted the efficacy of ESI in the treatment of patients with back and leg pain. Steroid medications do benefit some patients with radicular pain, but the benefit is often limited in duration, making efficacy difficult to prove over time. Steroids appear to speed the rate of recovery and return to function, however, allowing patients to reduce medication levels and increase activity while awaiting the natural improvement expected in most spinal disorders. Fluoroscopic verification of needle placement, with contrast injection, greatly improves steroid delivery while reducing risks. Although it is assumed that the benefit of steroids is related to their effect on inflammation, that remains unproven, and it is possible that benefit is gained through an unrecognized action. CONCLUSIONS: Randomized, controlled trials are needed to conclusively identify those patients most likely to benefit from ESI, and when and for how long. Until then, epidural steroids provide a reasonable alternative to surgical intervention in selected patients with back and/or leg pain, whose symptoms are functionally limiting. When appropriate goals are established and proper patients are selected, sufficient short-term benefit has been documented to warrant continued use of this tool. PMID- 15749620 TI - "Percutaneous vertebroplasty through a transdiscal access route after lumbar transpedicular instrumentation". PMID- 15749622 TI - Preemptive analgesia. PMID- 15749623 TI - Hormonal response to diets high in soy or animal protein without and with isoflavones in moderately hypercholesterolemic subjects. AB - Consumption of soy protein has been associated with altered risk of developing endocrine-regulated cancers. This study was designed to assess the independent effect of soy relative to animal protein and soy-derived isoflavones on circulating estrogen and androgen concentrations in postmenopausal women and older men. Forty-two subjects (> 50 yr) with low-density lipoprotein cholesterol levels of > or = 3.36 mmol/l were fed each of 4 diets in randomized order for 6 wk/phase. All food and drink were provided. Diets contained 25 g soy or common sources of animal protein/4.2 MJ containing trace or 50 mg isoflavones/4.2 MJ. At the end of each diet phase, concentrations of estrone sulfate, estrone, estradiol, testosterone, androstendione, dihydrotestosterone, dehydroepiandrosterone, and dehydroepiandrosterone sulfate were measured. In postmenopausal women, concentrations of estrone were higher and its precursor, dehydroepiandrosterone, lower after consuming the soy compared with animal protein diets (P = 0.0396 and 0.0374, respectively). There was no significant effect of isoflavones on any of the hormones measured. In older men, dehydroepiandrosterone sulfate concentrations were lower after consuming the isoflavone (P = 0.0106) and higher after soy, compared with the animal protein diets (P = 0.0118). These data suggest that relatively large amounts of soy protein or soy-derived isoflavones had modest and limited sex-specific effects on circulating hormone levels. PMID- 15749624 TI - Exploring vitamin and mineral supplementation and purported clinical effects in patients with small cell lung cancer: results from the Mayo Clinic lung cancer cohort. AB - Previous laboratory and pilot clinical trial data suggest that vitamin and/or mineral supplementation may prevent tumor growth in small cell lung cancer. However, rates of supplementation and their major purported clinical effects have never before been studied in patients with small cell lung cancer. This study was undertaken to explore associations between vitamin/mineral supplementation and survival and quality of life within a cohort of small cell lung cancer patients. This study focused on a small cell lung cancer patient cohort from a tertiary care medical center. Small cell lung cancer patients who responded to a follow-up questionnaire on vitamin/mineral use were included. Associations between vitamin/mineral use and both survival and quality of life (Lung Cancer Symptom Scale) were assessed. A total of 178 patients or their proxies responded to one or more vitamin/mineral questionnaires. One hundred seven (60%) were vitamin/mineral users of either multivitamins or other more specific vitamin/mineral supplements, and the rest were nonusers. Two different survival analyses were performed. In the first, median survival was 1.8 vs. 1.3 yr for vitamin/mineral users and nonusers, respectively. The relative risk of death was 0.63 (95% confidence interval [CI]: 0.43, 0.92; P = 0.02) in favor of vitamin/mineral use. After adjustment for multiple prognostic factors, including tumor stage, the relative risk for death was 0.65 (95% CI: 0.43, 1.00; P = 0.05). The second analysis was based on an alternative definition of vitamin/mineral use and showed only a trend to suggest an association between vitamin/mineral use and improved survival (P = 0.09). There were no significant improvements in quality of life in any of the analyses. Vitamin/mineral supplementation is common within this cohort of small cell lung cancer patients. These data suggest an association between vitamin/mineral supplementation and improved survival and point to a need for future studies on vitamin and mineral supplementation in small cell lung cancer patients. PMID- 15749625 TI - Serum carotenoids, retinol, and tocopherols, and colorectal cancer risk in a Japanese cohort: effect modification by sex for carotenoids. AB - To examine associations of serum carotenoids, retinol, and tocopherols with colorectal cancer risk, we conducted a case-control study nested within the Japan Collaborative Cohort Study. These micronutrients were measured in prediagnostic serum samples from 116 men and women who developed colorectal cancer during an 8 yr follow-up period and from 298 matched controls. In men, the higher level of serum total carotenoids was associated with a decreased risk: The multivariate adjusted odds ratio (OR) for the highest vs. the lowest tertile was 0.34 (95% confidence interval [CI] = 0.11-1.00; trend P over tertiles = 0.040). In women, the higher levels of alpha- and and total carotenoids were instead related to an increased risk: The corresponding ORs were 4.72 (95% CI = 1.29-17.3), 2.00 (0.70 5.73), and 2.47 (0.73-8.34), respectively (trend P = 0.007, 0.040, and 0.064, respectively). We also found a somewhat decreasing risk with increased serum retinol in all subjects and alpha-tocopherol in men: The ORs (95% CI) for the highest tertiles were 0.29 (0.11-0.78; trend P over tertiles = 0.010) and 0.29 (0.07-1.17; trend P = 0.098), respectively. The effects of some carotenoids on colorectal cancer risk may be modified by sex or by factors associated with sex, including smoking and drinking habits. PMID- 15749626 TI - Plasma uric acid levels in women with cervical intraepithelial neoplasia. AB - The objective of this study was to determine the association of plasma levels of uric acid, an endogenous antioxidant, in women with cervical intraepithelial neoplasia (CIN), while controlling for the confounding effects of human papillomavirus (HPV) infection, age, smoking, and use of oral contraception. Plasma-reduced and oxidized uric acid levels were determined in 650 women by high performance liquid chromatography, employing electrochemical technique. The findings demonstrated that 1) plasma-reduced uric acid (PRUA) levels in women with CIN (n = 311) were significantly lower (P < 0.05) compared with women in a control group (n = 339); 2) according to multiple logistic regression analysis, PRUA levels were negatively (P = 0.0113) and HPV infection were positively associated (P < 0.0001) with CIN, after controlling for the confounding effects of the studied factors; 3) according to multiple regression analysis, there was a 31% decrease in CIN risk for each incremental increase of 1mg/dl of PRUA; and 4) according to polychotomous logistic regression analysis, independent of HPV infection, PRUA level was inversely associated with the histopathological graded severity of CIN. We have previously reported decreased plasma levels of exogenous antioxidants, for example, vitamins C and E, in women with CIN independent of HPV infection. The data suggest that plasma deficiencies of several antioxidants in HPV-infected uterine cervical tissue may create an oxidative environment that renders the tissue susceptible to free radical damage. It may be speculated that chronic free radical-induced tissue damage in the context of persistent HPV infection may be involved in the pathogenesis of CIN. PMID- 15749627 TI - Pilot study: potential role of vitamin D (Cholecalciferol) in patients with PSA relapse after definitive therapy. AB - When local treatments for prostate cancer have failed, and prostate-specific antigen (PSA) rises in the absence of symptoms, there is little consensus as to the best management strategy. Calcitriol has been shown to prolong the doubling time of PSA in this context, but near-toxic doses are required. We investigated the effect of the nutrient vitamin D (cholecalciferol), a biochemical precursor of calcitriol, on PSA levels and the rate of rise of PSA in these patients. Fifteen patients were given 2,000 IU (50 microg) of cholecalciferol daily and monitored prospectively every 2-3 mo. In 9 patients, PSA levels decreased or remained unchanged after the commencement of cholecalciferol. This was sustained for as long as 21 mo. Also, there was a statistically significant decrease in the rate of PSA rise after administration of cholecalciferol (P = 0.005) compared with that before cholecalciferol. The median PSA doubling time increased from 14.3 mo prior to commencing cholecalciferol to 25 mo after commencing cholecalciferol. Fourteen of 15 patients had a prolongation of PSA doubling time after commencing cholecalciferol. There were no side effects reported by any patient. Further study is needed to confirm this finding and to explore the potential therapeutic benefit of nutrient vitamin D in prostate cancer. PMID- 15749628 TI - Possible mechanisms behind the differential effects of soy protein and casein feedings on colon cancer biomarkers in the rat. AB - In the present studies, several hypotheses were tested to explain previously reported differential effects of soy and casein on colon cancer biomarkers like cell proliferation, fecal fat, fecal bile acid, alkaline phosphatase, and magnesium excretion in rats. In Study 1, the effect of methionine, a limiting amino acid in soy protein and an amino acid that is thought to have a marked effect on colonic cell proliferation, was tested. It was concluded that methionine up to 1% in the diet had no effect on cell proliferation, using the 3H thymidine assay. The same study revealed that fecal alkaline phosphatase excretion is a good marker for colonic epithelial damage and fecal magnesium excretion is not. In Study 2, the hypothesis was tested that soy fractions enriched with isoflavones and saponins may increase fat excretion and so influence colonic cell proliferation in rats. It was indeed shown that soy protein isolate and an ethanolic extract from soy protein isolate slightly increased fecal fat excretion (up to 1.7-fold). However, fecal water bile acid and free fatty acid concentrations were decreased after feeding soy protein-based diets compared with casein, and no difference in fecal alkaline phosphatase excretion was observed. In Study 3, the lytic potential of soy saponins and the interaction between saponins and some lytic bile acids were tested in vitro. Data suggest a protective effect from soy saponins by reducing lytic activity of cholic acid. The overall conclusion is that soy protein compared with casein influences several colon cancer risk parameters, indicating a more protective rather than a stimulating effect on colon cancer risk. PMID- 15749629 TI - Resistant starch attenuates colonic DNA damage induced by higher dietary protein in rats. AB - Epidemiologic studies suggest that dietary complex carbohydrates are protective against colorectal cancer but dietary protein may increase risk. However, experimental data to support these relationships are scant. We have shown in rats that consumption of a high-protein (25% casein) diet for 4 wk resulted in a twofold increase in damage to colonocyte DNA compared with a low-protein (15% casein) diet. This was associated with thinning of the colonic mucous barrier and increased levels of fecal p-cresol. Addition of resistant starch as a high amylose maize starch to the diet increased cecal short-chain fatty acid pools and attenuated DNA damage, suggesting protection against genotoxic agents. In humans, this could translate to altered risk of colonic cancer. PMID- 15749630 TI - Chemopreventive effects of dietary flaxseed oil on colon tumor development. AB - Fatty acid composition of dietary fat, primarily the levels of omega-3 and omega 6 polyunsaturated fatty acids, has shown profound effect on colon tumor development in animal studies. Fats containing omega-6 fatty acids (for example, corn oil) enhanced and omega-3 fatty acids (for example, fish oil and mustard oil) reduced chemically induced colon tumors in rats. The purpose of this study was to investigate the effects of dietary flaxseed oil (containing alpha linolenic acid, an omega-3 polyunsaturated fatty acid) on azoxymethane-induced colon tumor in rats and how it compared with the dietary corn oil-treated group. Male Fischer rats, separated into 2 groups of 30, were assigned to the AIN-93M diet, which was supplemented with either 15% corn oil or 15% flaxseed oil. Carcinogenesis was initiated with subcutaneous injections of azoxymethane (15 mg/kg) once a week for three consecutive weeks. Thirty-five weeks after initiation, the rats were sacrificed under ether anesthesia. Blood was collected by cardiac puncture. The gastrointestinal tract was isolated and flushed with ice cold normal saline. The site, size, and number of tumors were recorded. The incidence and multiplicity of the tumors in the colon were determined. The fatty acid composition in the serum, colon, and tumors was estimated by using gas chromatography-flame ionization detection. Colon tumor incidence was found to be 100% and 54%, whereas multiplicity was found to be 3.1 and 0.7 tumors per rat in corn oil- and flaxseed oil-treated groups, respectively. Tumor size was significantly larger in the corn oil-treated group than in the flaxseed oil group. Colon and serum samples of the corn oil group showed an increase in the omega-6 fatty acid levels, whereas the flaxseed oil group exhibited an increase in the omega-3 fatty acid levels. The results indicate that dietary flaxseed oil, containing high levels of omega-3 fatty acids, is effective in preventing colon tumor development when compared with dietary corn oil containing omega-6 fatty acids in rats. PMID- 15749631 TI - Chemoprevention by Hippophae rhamnoides: effects on tumorigenesis, phase II and antioxidant enzymes, and IRF-1 transcription factor. AB - Fruits or berries of Hippophae rhamnoides (sea buckthorn), a rich source of vitamins A, C, and E, carotenes, flavonoids, and microelements such as sulfur, selenium, zinc, and copper, are edible and have been shown to protect from atopic dermatitis, hepatic injury, cardiac disease, ulcer, and atherosclerosis. However, its mechanism of action is not clear. We show that Hippophae inhibits benzo(a)pyrene-induced forestomach and DMBA-induced skin papillomagenesis in mouse. This decrease in carcinogenesis may be attributed to the concomitant induction of phase II enzymes such as glutathione S-transferase and DT-diaphorase and antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase in the mouse liver. This was accompanied by a remarkable induction of the transcription factor interferon regulatory factor-1 in the Hippophae-treated liver. Our results strongly suggest that Hippophae fruit is able to decrease carcinogen-induced forestomach and skin tumorigenesis, which might involve up-regulation of phase II and antioxidant enzymes as well as DNA binding activity of IRF-1, a known antioncogenic transcription factor causing growth suppression and apoptosis induction for its anticancer effect. PMID- 15749632 TI - Inhibition by allyl sulfides and crushed garlic of O6-methylguanine formation in liver DNA of dimethylnitrosamine-treated rats. AB - Garlic consumption is linked with lower incidences of certain cancers perhaps because garlic-derived allyl sulfides inhibit nitrosamine activation by cytochrome P450s. To help evaluate this view, effects of allyl sulfides on O6 methylguanine (O6MG) levels were examined in liver of rats injected with 20 mg/kg of liver carcinogen dimethylnitrosamine (DMN) and killed 3 h later. DNA was isolated and hydrolyzed, and O6MG/guanine ratios were determined by HPLC fluorescence. Mean inhibition of O6MG formation fell from 89% for 200 to 33% for 12 mg diallyl sulfide (DAS) per kilogram gavaged 18 h before DMN injection. Gavage of DAS 3 or 6 h (instead of 18 h) before DMN injection significantly reduced inhibitions. Mean inhibitions for diallyl disulfide, diallyl sulfoxide, and diallyl sulfone (75-100 mg/kg) gavaged 18 h before DMN were 39%, 72%, and 82%. In lung and kidney, DAS produced mean inhibitions of 98% and 74% compared with 89% in liver. When methylnitrosourea was injected instead of DMN, neither DAS nor DADS inhibited O6MG formation in liver DNA. Feeding 2.5% garlic for 7 days inhibited DMN-induced O6MG formation in liver DNA by 46%, similar to that expected from the estimated yield of allyl sulfides from garlic. Hence, dosing with DAS or feeding garlic may be useful chemopreventive strategies against nitrosamine-induced cancers. PMID- 15749633 TI - Citrus Reticulata blanco induces apoptosis in human gastric cancer cells SNU-668. AB - Citrus fruits have been known to reduce the proliferation of many cancer cells. The antiproliferative effects of Citrus reticulata Blanco (CR) extract, the immature tangerine peel, on human gastric cancer cell line SNU-668 were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6 diamidineo-2-phenylindole staining, terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay, reverse transcription-polymerase chain reaction expressions of BCL-2, BAX and CASP-3 genes, caspase-3 activity, and immunocytochemistry of caspase-3. From the results of the morphological and biochemical assays, CR (50 microg/ml) increased the apoptosis of human gastric cancer cells with typical apoptotic characteristics, including morphological changes of chromatin condensation and apoptotic body formation. CR (50 microg/ml) reduced the expression of BCL-2, whereas the expression of BAX and CASP-3 was increased compared with the control group. Furthermore, caspase-3 activity and caspase-3 protein expression in the CR-treated group was significantly increased compared with that in control group. These results suggest that CR may induce the apoptosis through the caspase-3 pathway in human gastric cancer cells. PMID- 15749634 TI - Bifidobacterium Lactis sp. 420 up-regulates cyclooxygenase (Cox)-1 and down regulates Cox-2 gene expression in a Caco-2 cell culture model. AB - Cyclooxygenases (Cox) -1 and -2 play important roles in gastrointestinal health; chronic overexpression of Cox-2 is associated with inflammatory and cancerous disease, whereas Cox-1 is expressed constitutively. We studied the effects of two probiotic (Bifidobacterium lactis sp. 420 and Lactobacillus acidophilus) and two control microorganisms (Escherichia coli and Salmonella enteritidis) and four microbial metabolites (acetate, butyrate, lactate and propionate) on the expression levels of the Cox isoforms in the enterocyte-like cell line Caco-2. Butyrate, which is anticarcinogenic, resulted in an 85% down-regulation of Cox-2 and a 37-fold increase in Cox-1 transcription. Propionate gave similar results (72% reduction of Cox-2, 23-fold induction of Cox-1), but lactate and acetate had no effect on Cox expression profile. Bifidobacterium sp. 420, which produces acetate and lactate but no butyrate or propionate, shared the Cox-1-increasing and Cox-2-silencing properties of butyrate and propionate, whereas L. acidophilus was similar to E. coli and S. enteritidis in having no effect on the Cox-1/Cox-2 ratio. For the first time, we therefore demonstrate evidence for a direct relationship between a probiotic bacterial strain and host Cox expression profile, suggesting that modulation of Cox expression may be an important factor in the potential anti-inflammatory and anticarcinogenic properties of some probiotics. PMID- 15749635 TI - Genistein induces apoptosis in T lymphoma cells via mitochondrial damage. AB - The soy isoflavone genistein has been identified as having antiproliferative and apoptotic effects on various malignant cell types derived from solid tumors. Because little information regarding the effect of genistein on hematopoietic malignancies is available, we undertook this study of T-cell lymphomas. We tested the effect of genistein on murine T-cell lines derived from thymic lymphomas induced by an oncogenic murine leukemia virus. When T lymphoma cells were treated with genistein concentrations of 15 microM and greater, it was observed that the percentage of viable cells was significantly reduced in a dose- and time dependent manner. The observed cell killing was found to be the result of apoptosis as detected by flow cytometric analysis of cells stained with annexin V and propidium iodide and assays for caspase-3 activation and DNA fragmentation. Cell staining with the mitochondrial specific dye JC-1 and detection of caspase-9 activation revealed that genistein produced mitochondrial depolarization as an early step in the induction of apoptosis. Bongkrekic acid inhibition of mitochondrial depolarization identified the mitochondria permeability transition pore (PTP) as a potential target of genistein activity. These results indicate that the induction of apoptosis by pharmacological concentrations of genistein in T lymphoma cells occurs via mitochondrial damage with the involvement of the PTP. PMID- 15749636 TI - Effects of fermentation products of pro- and prebiotics on trans-epithelial electrical resistance in an in vitro model of the colon. AB - Evidence from in vivo and in vitro studies suggests that the consumption of pro- and prebiotics may inhibit colon carcinogenesis; however, the mechanisms involved have, thus far, proved elusive. There are some indications from animal studies that the effects are being exerted during the promotion stage of carcinogenesis. One feature of the promotion stage of colorectal cancer is the disruption of tight junctions, leading to a loss of integrity across the intestinal barrier. We have used the Caco-2 human adenocarcinoma cell line as a model for the intestinal epithelia. Trans-epithelial electrical resistance measurements indicate Caco-2 monolayer integrity, and we recorded changes to this integrity following exposure to the fermentation products of selected probiotics and prebiotics, in the form of nondigestible oligosaccharides (NDOs). Our results indicate that NDOs themselves exert varying, but generally minor, effects upon the strength of the tight junctions, whereas the fermentation products of probiotics and NDOs tend to raise tight junction integrity above that of the controls. This effect was bacterial species and oligosaccharide specific. Bifidobacterium Bb 12 was particularly effective, as were the fermentation products of Raftiline and Raftilose. We further investigated the ability of Raftilose fermentations to protect against the negative effects of deoxycholic acid (DCA) upon tight junction integrity. We found protection to be species dependent and dependent upon the presence of the fermentation products in the media at the same time as or after exposure to the DCA. Results suggest that the Raftilose fermentation products may prevent disruption of the intestinal epithelial barrier function during damage by tumor promoters. PMID- 15749637 TI - Ursodeoxycholic acid can suppress deoxycholic acid-induced apoptosis by stimulating Akt/PKB-dependent survival signaling. AB - The nontoxic bile acid ursodeoxycholic acid (UDCA) is reported to be an anti apoptotic agent with efficacy against a variety of death stimuli including the cytotoxic bile acid deoxycholic acid (DCA). To gain insight into this anti apoptotic property, we tested UDCA for its ability to protect the colon carcinoma derived cell line HCT116 against DCA-induced apoptosis. We found that UDCA could suppress DCA-induced apoptosis in a time- and dose-dependent manner and that this effect correlated with Akt phosphorylation. Importantly, UDCA lost its ability to protect cells from DCA-induced cell death when Akt activity was suppressed genetically using a dominant negative Akt mutant or when PI3K activity was inhibited pharmacologically. These results suggest that UDCA can protect HCT116 cells against DCA-induced apoptosis by stimulating Akt-dependent survival signaling. PMID- 15749638 TI - Targeted cancer therapies. AB - The term "targeted cancer therapies" refers to treatment strategies designed to inhibit the product of an oncogene involved in the process of neoplastic transformation. Different categories of targeted therapies can be identified: 1) Therapies directed at oncogenes that are directly involved in the initiation of neoplastic transformation: the use of imatinib for the treatment of CML or GIST is the classical model in this subgroup. Single agent targeted therapies generally produce high response rates in this situation. 2) Therapies directed at oncogenes involved at a later stage of neoplastic transformation. These oncogenes contribute to tumor progression but not necessarily to the onset of malignant transformation. The use of trastuzumab for HER2-amplified breast adenocarcinoma is the classical model in this subgroup. These treatments are associated with low response rates when used as single agent therapy, whereas generally displaying a synergistic or additive effect with classical chemotherapy in models currently available. In contrast, when these targeted therapies are applied to tumor models where the targeted gene is present but not directly involved in the process of malignant transformation, no antitumor efficacy is generally observed. Recently, the identification of HER1 mutations in subsets of lung carcinoma as a predictive factor for response to gefitinib and erlotinib provided an example of how the empiric use of a targeted treatment may enable to identify new nosological entities. The present paper reviews examples of targeted cancer therapies and their results. PMID- 15749639 TI - A retrospective review of 15 years of radical radiotherapy with or without concurrent cisplatin and/or 5-fluorouracil for the treatment of locally advanced cervical cancer. AB - Radiation therapy is the standard of care treatment for locally advanced cervical cancer in the United States. In 1999 the addition of concomitant chemotherapy to radical radiotherapy became standard. The addition of cisplatin (CDDP) with or without 5-fluorouracil (5-FU) chemotherapy to radiation therapy was based on the near simultaneous reporting of five randomized, controlled clinical trials which all showed an improvement in survival with a magnitude of approximately 35%. The purpose of our study was to test the hypothesis that the addition of chemotherapy improved survival in our patients. We identified 291 patients treated with primary 'intent-to-cure' radiation therapy for locally advanced carcinoma of the cervix between 1985 and 2000. We analyzed patients using a stepwise Cox regression, including as possible predictors: clinical stage, age at diagnosis, use of concurrent chemotherapy with radiation and method of teletherapy delivery. We also examined survival as a function of CRT with a CDDP and/or 5-FU containing regimen using the Kaplan-Meier estimates of overall survival. The use of concurrent CDDP and/or 5-FU chemotherapy with radiation (CRT) was not associated with an increase in disease free survival (p=0.734) or overall survival (p=0.989). In this retrospective study there was no disease free or overall survival benefit from the addition of CDDP and/or 5-FU chemotherapy to radical radiotherapy for the treatment of locally advanced cervical carcinoma, although there was a trend favoring CRT. PMID- 15749642 TI - [Preclinical models of prostate cancer]. AB - Although prostate cancer is a major cause of cancer death and morbidity in Western countries, one major hindrance in the study of the biology of prostate cancer has been the limited number of laboratory models, compared with the number of models available for other neoplasms. For a long time, only three cell lines, namely LNCaP, PC3, and DU145, were routinely used to study prostate cancer in the lab. The success rate to establish cell lines from human prostate cancer tissues is low, in the 1% range. Currently, only about 10 prostate cancer cell lines are available, and many of them do not reproduce typical features of the human disease, like androgen receptor expression or prostate specific antigen (PSA) secretion. Spontaneous models in animals (including rat and dog) are practically not convenient for research purposes. Several in vivo models were artificially established by transforming prostate cells by potent oncogenes. Other models were developed by injecting prostate cancer cell lines into the prostate (orthotopic model), the vessel, or the bones of immuno-deficient mice, to mimic localized and metastatic prostate cancer. This was successfully done with the MDA PCa2b cell line. This cell line was also used to generate an in vitro model of bone metastases by a co-culture system with osteoblasts. This model allows to study the paracrine cross-talk between the two cell compartments and the resulting molecular modifications. The objective of the present article is to review the currently available model systems of prostate cancer. PMID- 15749641 TI - [Hypoxia-inducible factor 1: regulation, involvement in carcinogenesis and target for anticancer therapy]. AB - Hypoxia-inducible factor-1 is a heterodimer made up of an oxygen-regulated HIF1alpha subunit and a constitutively expressed HIF1beta subunit. Among the 70 target genes of HIF-1 known so far, several are involved in angiogenesis, erythropoiesis, cell proliferation, cell viability, and glucose and iron metabolisms. Intratumoral hypoxia or genetic alterations can lead to HIF-1 alpha over-expression. HIF-1 over-expression has been associated with an increased patient mortality rate in many cancer types. Also, in vitro suppression of hif1alpha gene expression has been shown to be efficient in tumour growth repression. During the past five years, drugs able to indirectly inhibit HIF1 activity have been rationally or empirically developed. Some are currently evaluated in clinical trials, but further work has still to be undertaken to rationally identify new specific inhibitors of HIF1 and to test their efficacy as anticancer therapeutics. This review focuses on HIF1 regulation, HIF1 involvement in tumour promotion, the different HIF-1 inhibitors currently tested and their mechanisms of action. PMID- 15749640 TI - Expression of class III beta tubulin in non-small cell lung cancer is correlated with resistance to taxane chemotherapy. AB - This study determined the prevalence and the prognostic value of the expression of microtubule components in tumors of 19 patients with non small cell lung cancer receiving taxane-based regimens. Patient samples were stained with antibodies directed against total beta tubulin, classes I, II and III beta tubulin isotypes, delta2 alpha tubulin, tau protein, and the P-gp protein involved in the classical multidrug resistance phenotype. All tumors were stained with pan-beta tubulin antibody and class I tubulin isotype. A majority of the tumor samples expressed class II and class III, although the percentage of positive cells varied significantly between tumors. delta2 alpha tubulin, tau protein and Pgp protein were found in only one tumor sample each. Progression free survival was shorter (41 days) in patients whose tumors expressed high levels of class III tubulin isotype in comparison to patients with low levels (288 days, p = 0.02). There were 2 responses to chemotherapy among 9 patients (22%) with high levels of class III tubulin vs. 6 among 10 patients (60%) with low levels of expression (Fisher exact test: p = 0.11). These data suggest that high expression of class III tubulin by tumor cells is associated with poor prognosis in patients with NSCLC receiving a taxane-based regimen. PMID- 15749643 TI - [Hormone therapy in breast cancer: the end of tamoxifen?]. AB - The rise of aromatase inhibitors changes our habits in hormonal treatment in breast cancer. In premenopausal women tamoxifen with ovarian suppression remains reference treatment although the succession of the two treatments has not been explored as far as treatment duration is concerned. In post menopausal women the use of AI in the first line treatment gives a greater chance of response and a longer response duration than tamoxifen with a good tolerance profile. When both treatments are used, total duration of endocrine therapy (time to chemotherapy) is significantly longer for patients who receive AI in first line than for patients who receive tamoxifen first. In adjuvant treatment, tamoxifen and ovarian suppression remain references in premenopausal patients; therefore trials using aromatase inhibitors with LH-RH agonists are running. In post menopausal women aromatase inhibitors are challenging tamoxifen. Three large trials proved their superiority over tamoxifen. In the adjuvant setting AI may represent a beneficial approach, they can be used up front or in sequence with tamoxifen. The ideal combination or sequence of therapies requires investigations. At last duration of hormonal treatment remains an open deal. PMID- 15749644 TI - [Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy for peritoneal colorectal carcinomatosis: a newly validated standard whose contribution remains to be assessed]. AB - Peritoneal carcinomatosis represent an advanced form of colorectal cancer but different of others metastatic pathways. However carcinomatosis was associated with stage M in the TNM classification, because of a very poor prognosis, actually resulting despite new chemotherapeutic agents of a single year of survival. In an estimated 30% of patients, no other tumour location can be found. Usually complete tumour resection can not be perform, resulting in the best time with a R1 procedure associated with microscopic residual disease. This is to eradicate microscopic residual disease that hyperthermic intraperitoneal chemotherapy (HIPEC) was proposed. First results of single unit published reported survival after 5 years and that the most important prognosis factor of survival was complete resection of the carcinomatosis (R1 resection). A phase III study publish in 2003 in the Journal of Surgical Oncology by the Zoetmulder's team confirm the phase II results and offer for the first time a factual impact with a level II of demonstration. Between 1998 and 2001, 105 patients were randomly assigned to receive either standard treatment consisting of systemic chemotherapy (fluorouracil-leucovorin) or aggressive cytoreduction (incomplete for some patients) with HIPEC (mitomycin C during 90 minutes) followed by the same systemic chemotherapy regime. In the HIPEC arms, mortality was at 8 % and the median hospital admission was near one month. After a median follow-up period of 21 months, the median survival was 12 months in the standard therapy arm and 22 months in the HIPEC arm (p = 0.032). These results confirm prior reported non randomised study reporting median survival of 12 to 40 months. We had recently published the results of a phase II study including only patient with a R1 resection associated with HIPEC with oxaliplatin. This study had include 24 patients with an overall survival of 65% at 3 years. These survival result was the most prolonged never reported in the treatment of peritoneal colorectal carcinomatosis. HIPEC can be considered as a new standard even the exact place in clinical practice had to be more define because of alls recent progress are publish giving an important number of publications during the 2003-2004 years, some of them coming from French groups. PMID- 15749645 TI - [Standards, Options and Recommendations for the management of ductal carcinoma in situ of the breast (DCIS): update 2004]. AB - The " Standards, Options and Recommendations " (SOR) project, started in 1993, is a collaboration between the Federation of French Cancer Centres (FNCLCC), the 20 French cancer centres, and specialists from French public universities, general hospitals and private clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. Objectives : To update the Standards, Options and Recommendations clinical practice guidelines for the management of ductal carcinoma in situ of the breast (DCIS). Methods : The working group identified the questions requiring up-dating from the previous guideline. Medline(r) and Embase(r) were searched using specific search strategies from year 1996 to year 2003. In addition several Internet sites were searched in October 2002. Results : Clinical guidelines have been defined for the management of diagnosis, treatment, follow-up, and treatment of recurrence of DCIS. The issue of hormone replacement therapy has also been addressed in the context of DCIS. PMID- 15749647 TI - [Axillary lymphocele after axillary dissection or sampling of sentinel lymph node in breast cancer]. AB - The goal of this study was to evaluate the rate of seromas after axillary dissection or sentinel lymph node biopsy. This is a prospective study based upon a series of 229 patients. Among those 229 patients, 179 had an axillary dissection and 50 had a sentinel lymph node biopsy. In the axillary dissection group, 40% of patients developed a seroma. The maximum number of aspirations needed was 8. In the sentinel lymph node group, 18% of patients developed a seroma that never recurred after a single aspiration. Seromas are still a very frequent complication after axillary dissection. The sentinel lymph node biopsy has helped to reduce the rate of axillary seroma, and the number of aspirations needed to evacuate them. PMID- 15749646 TI - [Analysis of the medical activity related to cancer in a network of multidisciplinary hospitals using claims databases, the reseau Concorde Oncology Network]. AB - Recently, to answer patients, caregivers and professionals needs, the "Plan Cancer" has been presented by the French Government. This plan is intended to improve quality of care in cancer patients and finally, patients' survival and quality of life. This planned strategy stresses the importance of organized interactions between hospitals and between the various health professionals. Measuring the number of patients with cancer and the activity related to cancer in large networks of multidisciplinary hospitals has became a real challenge in France for organizational, quality of care and economic reasons. Many University Hospitals in France have chosen to face this question by using the French DRG based information system called PMSI. It allows estimating the proportion of hospital stays concerned by cancers that are identified with algorithms based on ICD 10. However, French databases of hospital discharges do not allow patients identification. We collected data on hospital stays and patients in a subset of an organized network focused on cancer care and composed of 55 public or private hospitals in the Rhone-Alpes area. We used these data to estimate the number of patients who had been hospitalized within the network in 2000. Approximately 110,000 hospital stays were related with a diagnostic of cancer, corresponding to a number of patients within a range of 30345 to 35700. In absence of communicating files between hospitals, claims databases are an interesting source of information for cancer burden. The recent implementation of a procedure allowing the linkage of data concerning each patient should permit better estimates in the future. The main limitation will remain the possibility of a hospital to participate to more than one network. PMID- 15749648 TI - [Role of an exclusive concomitant radio-chemotherapy treatment in non operable esophageal cancer: results of a 10-year experience in Antoine-Lacassagne Center]. AB - Treatment of non operable esophageal cancer still remains debatable. To date, radio-chemotherapy treatment could be considered as a standard, offers to the patients real hope and a new area of management in this unfavourable cancer. The aim of the present study was to report retrospectively a 10 year experience in concomitant radio-chemotherapy primary treatment in non operable esophageal cancer patients in Antoine Lacassagne anti cancer Center. Between January 1989 and June 1997, 63 consecutive, previously untreated patients with squamous cell carcinoma of the esophagus and who were inoperable for various reasons were majoritably treated with cisplatin (70 mg/m2) at J1 plus 5-fluorouracil (800 mg/m2/d) from J1 to J5 every 3 weeks (78% of patients) concomitantly with external beam radiotherapy (2 types). Two other chemotherapy regimens has been also used. Seventy-five percent (47/63) of the patients received the stipuled concomitant radio-chemotherapy dose. Neutropenia in the form of WHO grade 3-4 : 27% (17/63) was observed, grade 3-4 anemia and thrombopenia in 16 (26 %) and in 9 (15%) patients, respectively, grade 3-4 emesis in 6 % (4/63), grade 3-4 mucositis in 10 % (6/63). On 47 patients with accessible responder status, 18 of them presented a complete response, 20 a partial response, 6 a stable disease and 3 a progressive disease. The median follow up was 7 years. The median overall survival was 9.6 months with 11% estimated to be alive after 5 years. Combined treatment with cisplatin 5-fluorouracil and radiotherapy for inoperable cancer of the esophageal is relatively well tolerated and reasonably efficacious in a very selected group of patients. PMID- 15749650 TI - [How to deal with non conventional therapies?]. PMID- 15749649 TI - [Treatment of childhood Burkitt lymphoma according to LMB89 protocol in Casablanca]. AB - During the two last decades, the prognosis of children with Burkitt lymphoma has improved dramatically. Treating patients with Bukitt lymphoma in countries with limited resources is a challenge. We report our results in a serie of 95 children with Burkitt lymphoma treated between September 1990 and December 2000 according to SFOP LMB89 protocol. The median age was 45 months (range 8 months, 18 years). Seventy three percent of patients had abdominal tumor and 10% had maxillary tumor. According to Murphy classification, one patient had stage I, 17 patients stage II, 60 patients stage III and 17 patients stage IV. When considering the LMB prognosis groups, 1 patient was in group A, 83 were in group B and 11 were in group C. 73 patients were evaluables for treatment results. Complete remission was achieved in 50 patients, of whom 6 relapsed. 18 patients died from early treatment toxicity. The 5 years relapse free survival rate was 56%. It was at 100%, 84%, 52% and 38% for stage I, II, III and IV respectively. These results are below what is expected with this protocol. Improvement of supportive-care is the main condition to reach western results. PMID- 15749651 TI - Two novel mutations, L490R and V561X, in transferrin receptor 2 in Japanese patients with hemochromatosis. PMID- 15749652 TI - Telomere length and hTERT expression in patients with acute myeloid leukemia correlate with karyotypic instability. PMID- 15749653 TI - A green tea component, catechin, rapidly induces apoptosis of myeloid leukemic cells via modulation of reactive oxygen species production in vitro and inhibits tumor growth in vivo. PMID- 15749654 TI - Additive effects in the induction of apoptosis in leukemic cells by WT1 and BCR ABL specific siRNA. PMID- 15749655 TI - Baseline disease risk and response to therapy in chronic myeloid leukemia: necessary predictive tools then and now. PMID- 15749656 TI - Immunohistochemical profiling of homogeneously treated de novo tumors with nodal presentation on tissue micro-arrays. PMID- 15749657 TI - The development of more than one histologic type of lymphoma in the same patient is frequent and confers a worse prognosis. PMID- 15749658 TI - Long-term bone marrow culture data is the most powerful predictor for peripheral blood progenitor cell mobilization in healthy donors. PMID- 15749659 TI - Why do humans need two types of transferrin receptor? Lessons from a rare genetic disorder. PMID- 15749660 TI - The future of anticoagulation clinics: a journey to thrombosis centers? PMID- 15749661 TI - Two novel mutations, L490R and V561X, of the transferrin receptor 2 gene in Japanese patients with hemochromatosis. AB - BACKGROUND AND OBJECTIVES: The low prevalence of the C282Y mutation of the HFE gene in Japan means that the genetic background of hemochromatosis in Japanese patients remains unclear. In a previous report, we showed that 3 patients from one family had an AVAQ 594-597 deletion of the transferrin receptor (TfR2) gene. This suggests that the TfR2 gene is involved in hemochromatosis in Japanese patients. DESIGN AND METHODS: Nine patients clinically diagnosed with hemochromatosis were included in the study. DNA was extracted from whole blood samples collected with informed consent. The HFE and TfR2 genes were analyzed by sequencing the coding region and splicing sites. RESULTS: There were no mutations in the HFE gene. In the TfR2 gene, 2 novel mutations, 1469T->G (L490R) and 1665delC (V561X), were found in 2 patients. A known variation, 714C-> (I238M), was also found in the patient with L490R. The patient homozygous for both L490R and I238M presented with a mild manifestation of hemochromatosis at the age of 41 years. His liver was cirrhotic with parenchymal iron deposits and the result of a glucose tolerance test was compatible with diabetes mellitus. The patient homozygous for V561X had severe iron overload with the triad of cirrhosis, diabetes mellitus and skin pigmentation at the age of 58 years. INTERPRETATION AND CONCLUSIONS: Taken together with the previous report, 5 of our 12 patients with hemochromatosis manifesting in middle age had mutations in the TfR2 gene. Thus, TfR2 plays a role in the pathogenesis of hemochromatosis in Japan. PMID- 15749662 TI - Telomere length and hTERT expression in patients with acute myeloid leukemia correlates with chromosomal abnormalities. AB - BACKGROUND AND OBJECTIVES: Acute myeloid leukemia (AML) is a malignant, genetically heterogenous disorder characterized by uncontrolled growth of immature myeloid cells. The aim of this study was to analyze whether telomere length and/or hTERT expression are correlated with clonal chromosomal aberrations in AML. DESIGN AND METHODS: Telomere length in mononuclear cells derived from 137 previously untreated patients with >or= 80% blasts was analyzed by flow fluorescent in situ hybridization. Results were expressed in telomere fluorescence Units (1 TFU=1 kb). The expression of hTERT, including its different splice variants was studied by reverse transcription-polymerase chain reaction. RESULTS: Age-adjusted telomere length in AML patients was significantly reduced as compared to in matched controls, consisting of peripheral blood granulocytes from healthy individuals (0-90 years) (median: -2.5 TFU; p<0.001). Patients with an aberrant karyotype had significantly shorter telomeres than patients with a normal karyotype (median -3.0 vs. -2.3 TFU; p=0.03). The shortest telomeres were found in patients with multiple aberrations (median -3.7 TFU; p=0.03). hTERT expression was found to be correlated with chromosomal abnormalities as well as with the detection of functional hTERT splicing variants. INTERPRETATION AND CONCLUSIONS: These findings suggest an important role of intense telomere loss in the development of genetic instability during the pathogenesis of AML. It is assumed that critical telomere shortening in AML blasts could lead to telomerase activation and therefore prevent blasts from replicative senescence, one possible mechanism for clonal selection and disease progression. Therefore, telomere length might serve as a prognostic marker for AML patients. These findings would have to be confirmed in large, prospective studies. PMID- 15749663 TI - Catechin, a green tea component, rapidly induces apoptosis of myeloid leukemic cells via modulation of reactive oxygen species production in vitro and inhibits tumor growth in vivo. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to investigate the possibility of green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG) as a novel therapeutic agent for the patients with myeloid leukemia. DESIGN AND METHODS: We investigated the effects of EGCG on the induction of apoptosis in leukemic cells in vitro and in vivo. We further examined the molecular mechanisms of EGCG-induced apoptosis in myeloid leukemic cells. RESULTS: EGCG rapidly induced apoptotic cell death in retinoic acid (RA)-resistant acute promyelocytic leukemia (APL), UF-1 cells within 3 h. EGCG induced apoptosis in UF-1 cells was in association with the loss of mitochondrial transmembrane potentials (Deltapsim) and activation of caspase-3 and -9. Elevation of intracellular reactive oxygen species (ROS) production was also demonstrated during EGCG induced apoptosis of UF-1 as well as fresh myeloid leukemic cells. In NOD/SCID mice transplanted with UF-1 cells, EGCG effectively inhibited tumor growth in vivo, and the number of mitoses among the cells significantly decreased in comparison to that of control mouse cells. INTERPRETATION AND CONCLUSIONS: In summary, EGCG has potential as a novel therapeutic agent for myeloid leukemia via induction of apoptosis mediated by modification of the redox system. PMID- 15749664 TI - WT1 and BCR-ABL specific small interfering RNA have additive effects in the induction of apoptosis in leukemic cells. AB - BACKGROUND AND OBJECTIVES: The Wilms' tumor gene (WT1) is aberrantly over expressed in leukemic cells. Therefore, we wanted to study the effect of small interfering (siRNA) targeting WT1 in leukemic cells and normal CD34-positive cells with regard to proliferation, induction of apoptosis, and cell differentiation. Furthermore, we wanted to evaluate whether the additional use of BCR-ABL siRNA could increase the anti-leukemic effects of WT1 siRNA in chronic myeloid leukemia (CML) cells. DESIGN AND METHODS: We measured WT1 expression by reverse transcription polymerase chain reaction (RT-PCR) in various cell lines and in leukemic cells from patients, then transfected the cells with WT1-specific and BCR-ABL-specific siRNA before carrying out microarray analysis. We used the tunnel assay to measure apoptotic cells. RESULTS: We observed a reduction of WT1 gene expression, measured by real-time RT-PCR, in all studied cell lines: K-562, Kasumi-1, MV 4-11 and NB-4, as well as in cells of AML and CML patients. The results also demonstrated that WT1 siRNA significantly induced apoptosis and inhibited proliferation in MV4-11 cells, NB-4 cells, Kasumi-1 cells (p<0.01) and in K-562 cells (p<0.02) versus controls. In normal CD34-positive cells, the proliferation was only slightly inhibited (by about 20%) and no induction of apoptosis was found. Combined transfection with WT1 and BCR-ABL siRNA together in K-562 cells increased the inhibition of the rate of proliferation and the rate of induced apoptosis compared to transfection with BCR-ABL siRNA or WT1 siRNA alone (p<0.01). We found that most genes involved in cell signaling and protein metabolism were regulated by the WT1 gene in K-562 cells in a microarray analysis. INTERPRETATION AND CONCLUSIONS: In conclusion, WT1 might be a suitable target for new therapeutic strategies using siRNAs in leukemic cells. PMID- 15749666 TI - Identification of outcome predictors in diffuse large B-cell lymphoma. Immunohistochemical profiling of homogeneously treated de novo tumors with nodal presentation on tissue micro-arrays. AB - BACKGROUND AND OBJECTIVES: Patients with diffuse large B-cell lymphoma (DLBCL) could benefit from integration of well-established bioclinical prognostic factors with new tools - such as micro-arrays - exploring aberrant gene and/or protein expression. DESIGN AND METHODS: Tissue micro-arrays (TMA) were constructed for the paraffin blocks of 68 patients with de novo DLBCL with nodal presentation, who underwent MACOP-B and were provided with complete clinical information. TMA were tested with specific antibodies against CD10, CD20, CD30, CD79a, CD138, Bcl 2, Bcl-6, IRF4, and IRTA1. RESULTS: The following phenotypic subclassification was made: a) CD10 +/Bcl-6 + or Bcl-6+/IRF4 +, but Bcl-2-/CD30-/CD138-- suggesting B-cells gathering/leaving the germinal center (group 1; n=36); b) Bcl-2+/CD10 /Bcl-6- and CD30+ or CD138+ corresponding to putative non-germinal center B-cells with features of activation or plasmablastic/plasmacellular differentiation (group 2; n=17); c) CD30-/CD138- with extensive Bcl-2 positivity and variable CD10, Bcl-6 and IRF4 combinations (group 3; n=15). Mean IPI scores were 0.6, 1.9 and 1.1 for groups 1, 2 and 3, respectively (p= 0.001). Complete remission (CR) rates were 89%, 53% and 73% (p= 0.015). The 3-year relapse-free survival (RFS) rates are 86%, 41% and 63% (p=0.001) and 42-month overall survival (OS) rates are 91%, 38% and 66% (p=0.0002). INTERPRETATION AND CONCLUSIONS: The present TMA based study suggests an immunophenotypic profiling system for patients with de novo DLBCL that seems to provide additional prognostic information and contributes to the existing debate on the identification of suitable immunohistochemical surrogates of gene expression profiling results. PMID- 15749665 TI - The impact of the combination of baseline risk group and cytogenetic response on the survival of patients with chronic myeloid leukemia treated with interferon alpha. AB - BACKGROUND AND OBJECTIVES: This study was aimed at examining major cytogenetic response (MCR) as a valid predictor of the course of chronic myeloid leukemia (CML) and at assessing the survival of CML patients treated with interferon alpha (IFN) in dependence on the combination of MCR (yes or no) with the baseline risk group of the New CML score. MCR was defined as a reduction of Philadelphia chromosome-positive bone marrow cells to 73 Kg, G-CSF schedule and results of LT-BMC, although in the multivariate analysis only the number of CFU-GM obtained after LT-BMC showed a significant influence (p<0.001). INTERPRETATION AND CONCLUSIONS: These results confirm the interindividual variation in HPC mobilization among healthy subjects, with LT-BMC counts being the most reliable predictor, expressing the behavior of the immature progenitors and their relationship with the microenvironment. PMID- 15749669 TI - Stem cell plasticity: time for a reappraisal? AB - In recent years an increasing number of publications have claimed that adult mammalian stem cells (SC) may be capable of differentiating across tissue lineage boundaries and that this plasticity may represent a novel therapeutic strategy for tissue regeneration. However, after a first phase of excitement, the issue of somatic SC plasticity remains controversial and the therapeutic perspectives are still elusive. In this review, we examine the general mechanisms which govern the function of SC, the identification and functional characterization of adult SC of different tissues and their putative capacity to transdifferentiate into mature cells of different origin. The potential clinical applications of adult SC for regenerative medicine are also discussed in each chapter. The method employed for preparing this review was the informal consensus development. Members of the Working Group on SC met four times and discussed the single points, previously assigned by the Chairman (S.T.), in order to achieve an agreement on different opinions and approve the final manuscript. All the authors of the present review have been working in the field of SC and have contributed original papers to peer reviewed journals. In addition to the authors' own work, the present review examines articles published in journals covered by the Science Citation Index and Medline. PMID- 15749670 TI - Molecular monitoring of childhood acute lymphoblastic leukemia using antigen receptor gene rearrangements and quantitative polymerase chain reaction technology. AB - The use of minimal residual disease (MRD) measurement as a surrogate marker of molecular response to treatment can potentially improve the evaluation of treatment response and enable estimates of the residual leukemic cell burden during clinical remission, thereby improving the selection of therapeutic strategies and, possibly, long-term clinical outcome. The most specific and sensitive methods for MRD monitoring currently available are polymerase chain reaction amplification of rearranged immunoglobulin and antigen-receptor genes, and flow cytometric detection of aberrant immunophenotypes. Several retrospective studies have demonstrated the strong association between MRD and risk of relapse in childhood acute lymphoid leukemia (ALL), irrespective of the methodology used. The promising results on the predictivity of MRD evaluation at the end of induction treatment has challenged the need for a new definition of remission. There is now urgent need to incorporate MRD data into clinical studies, properly designed to address treatment questions, in order to explore whether a better tailored treatment would result in further improvement in cure rates for children with ALL. However, several critical issues must be resolved before MRD determinations can be routinely considered in clinical decision making. PMID- 15749671 TI - Chronic lymphocytic leukemia: novel prognostic factors and their relevance for risk-adapted therapeutic strategies. AB - BACKGROUND AND OBJECTIVES: Many years ago it was established that prompt treatment of early stage chronic lymphocytic leukemia (CLL), the stage at which almost two-thirds of CLL patients present, has no benefit over a management of watching and waiting, then treating progression. However, this fact was based on series treated ineffectually with chlorambucil, which were not stratified according to prognostic markers. DESIGN AND METHODS: The prognosis and clinical course of CLL are heterogeneous. While some patients may have a normal life expectancy without requiring treatment, others die of drug-resistant disease as early as within two years of presentation. However, unlike the situation in non Hodgkin's lymphoma, there is no standard Prognostic Index that can be used to group patients with CLL according to likely outcome or to guide treatment. RESULTS: A number of clinical and biological factors of prognostic relevance, which may add to the classical assessment provided by the staging systems, have been identified. These include clinical characteristics, such as age, gender and performance status, and laboratory parameters reflecting the tumor burden or disease activity, such as lymphocyte count, lactate dehydrogenase (LDH) increase, bone marrow infiltration pattern or lymphocyte doubling time. Recently more informative prognostic parameters have been identified: serum markers such as soluble CD23, b2-microglobulin or thymidine kinase and genetic markers of tumor cells, such as genomic aberrations, gene abnormalities (p53, ATM), the mutation status of the variable segments of the immunoglobulin heavy chain genes (IGVH) or surrogate markers for these factors, such as CD38 and ZAP-70. INTERPRETATION AND CONCLUSIONS: From the clinician's perspective the importance of this new knowledge is how it affects treatment. It is now possible to produce molecular remissions even in advanced disease using combinations of purine analogs and monoclonal antibodies. Moreover, potentially curative therapeutic modalities such as autologous and allogeneic stem cell transplantation are becoming safer. Clinical trials of effective treatment stratified by more reliable prognostic markers are surely now warranted. PMID- 15749672 TI - Differential expression of vascular endothelial growth factor and its receptors in hematopoietic and fatty bone marrow: evidence that neuropilin-1 is produced by fat cells. AB - Vascular endothelial growth factor (VEGF), its receptors (VEGFR-1, VEGFR-2) and neuropillin-1 (NRP-1) are expressed at variable levels in bone marrow. NRP 1expression is higher in fatty bone marrow than in hematopoietic marrow. Adipocytes are responsible for NRP-1 expression suggesting that they may play a role in hematopoiesis by producing NRP-1 or that NRP-1 may regulate adipocyte activity. PMID- 15749673 TI - Decreased plasma endothelin-1 levels in children with sickle cell disease treated with hydroxyurea. AB - Plasma endothelin-1 (ET-1) is elevated in patients with sickle cell disease (SCD). Hydroxyurea (HU) is the only drug with demonstrated clinical efficacy in SCD. Here we show that treatment with HU results in a decreased concentration of circulating ET-1 which is not correlated with the HU-induced increase in HbF level. Blunting of the ET-1 vasoconstrictive stimulus could contribute to the beneficial effects of HU. PMID- 15749674 TI - Changes in expression of WT1 isoforms during induced differentiation of the NB4 cell line. AB - The levels of expression of WT1 gene and WT1+17AA isoforms rapidly decreased during the differentiation of NB4 cells induced by all-trans retinoic acid; this decrease was conversely related to the dynamic changes of CD11b positive cells, indicating that the abnormally high expression of WT1 gene and WT1+17AA isoforms was associated with a block of NB4 cell differentiation. PMID- 15749675 TI - Expression of polycythemia rubra vera-1 decreases the dependency of cells on growth factors for proliferation. AB - An increase in the level of polycythemia rubra vera -1 (PRV-1) mRNA has been reported in some myeloproliferative disorders. We have studied the effects of PRV 1 on cell proliferation and cell survival. In cell growth assays, the number of heterologous cells expressing PRV-1 increased faster than sham-transfected cells, a difference that was more pronounced in serum-free media. Even after 5 days of exposure to serum-free media, cells expressing PRV-1 continued to proliferate, whereas the control cells ceased to proliferate. We conclude that PRV-1 is a pro proliferation molecule, and hypothesize that its overexpression may have a role in the pathogenesis of myeloproliferative disorders. PMID- 15749676 TI - Neutrophil polycythemia rubra vera-1 expression in classic and atypical myeloproliferative disorders and laboratory correlates. AB - The current study of 153 subjects with both classic and atypical myeloproliferative disorders suggests that neutrophil polycythemia rubra vera-1 (PRV-1) over-expression is a non-specific feature of clonal myeloproliferation that displays significant correlation with leukocyte alkaline phosphatase score. These observations undermine the utility of the PRV-1 assay as a diagnostic test of additional value. PMID- 15749677 TI - Mutation of the prothrombin gene and thrombotic events in patients with polycythemia vera or essential thrombocythemia: a cohort study. AB - The association between a prothrombin mutation and the risk of thrombosis was analyzed in 214 patients with polycythemia vera or essential thrombocythemia. The rate for venous thrombotic events was 14.7/100 patient-years in patients with the prothrombin mutation compared to 0.8 in patients without the mutation (rate ratio 17.5). PMID- 15749678 TI - A pilot study of low-dose subcutaneous alemtuzumab therapy for patients with hemotherapy-refractory chronic lymphocytic leukemia. AB - Subcutaneous low-dose alemtuzumab (10 mg t.i.w. for 18 weeks) induced a 50% response rate, including 25% complete response, in 16 patients with refractory chronic lymphocytic leukemia (CLL) patients. The responses were substantial even in patients with unfavorable cytogenetics, fludarabine/rituximab refractoriness, Rai stage IV, previous infections, and age over 65 years. Subcutaneous low-dose alemtuzumab is effective in poor prognosis B-CLL, and has a particularly favourable toxicity profile. PMID- 15749679 TI - Low frequency of exon 3 PTPN11 mutations in adult de novo acute myeloid leukemia. Analysis of a consecutive series of 173 patients. AB - A total of 173 samples obtained from adult patients with de novo acute myeloid leukemia (AML) were assayed for exon 3 PTPN11 mutations by single strand conformation polymorphism (SSCP) analysis and direct sequencing. Only three monocytic leukemias had point mutations (1.73%). PMID- 15749681 TI - The t(12:21) is underrepresented in childhood B-lineage acute lymphoblastic leukemia in Kerala, Southern India. AB - t(12;21) (TEL/AML1) is the most common genetic event in childhood B-cell acute lymphoblastic leukemia (B-ALL) in Western countries. Samples from 42 children with ALL in Kerala were tested by reverse transcription polymerase chain reaction for TEL/AML1, t(1;19) and t(4;11). Only 2 out of 42 (4.8%) cases were positive for the TEL/AML1, and t(1;19) and t(4;11) were not detected. We conclude that the incidence of TEL/AML1 is lower in the Indian population. PMID- 15749680 TI - Tetraploidy or near-tetraploidy clones with double 8;21 translocation: a non random additional anomaly of acute myeloid leukemia with t(8;21)(q22;q22). AB - We report on 6 patients with tetraploidy or near-tetraploidy acute myeloid leukemia (AML) with double t(8;21) (q22;q22) and review the literature on cases with the same cytogenetic abnormalities. Some common features were revealed by this analysis. PMID- 15749682 TI - Family clustering of blood cancers as a risk factor for lymphoid neoplasms. AB - Family aggregation of cancer was significantly more common among 588 incident cases with lymphoid neoplasms than among 631 controls (OR: 1.4; 95%CI= 1.1-1.8, p value=0.004). This association was of particular relevance among cases of multiple myeloma and chronic lymphocytic leukemia, with a 2-fold increased risk, the latter also showing an almost 4-fold increased risk of family aggregation of hematologic cancers. PMID- 15749683 TI - Homozygosity for a Thr575Met missense mutation in the catalytic domain associated with factor XI deficiency. AB - In this study we investigated an asymptomatic 55-year-old Lebanese woman with factor XI deficiency. The F11 gene was analyzed and a cross reacting material positive (CRM+) variant, Thr575Met, was identified in homozygosity in the proband, and in heterozygosity in four of her siblings. PMID- 15749684 TI - Plasma soluble fibrin monomer complex is a useful predictor of disseminated intravascular coagulation in neonatal sepsis. AB - Disseminated intravascular coagulation (DIC) is a major factor influencing mortality in neonatal sepsis.(1) Clinical trials have supported the use of antithrombin and activated protein C supplementation in DIC associated with sepsis. PMID- 15749685 TI - Five prothrombotic polymorphisms and the prevalence of premature myocardial infarction. AB - We studied 5 functional hemostatic polymorphisms in 281 patients with premature myocardial infarction and in 530 control subjects. The role of these polymorphisms when analyzed independently is small, if any. However, the simultaneous combination of factor XIII and prothrombin polymorphisms exacerbated the risk. (OR=12.12; p=0.028). Moreover, combinations of factor V Leiden with prothrombin, and factor XII with prothrombin polymorphisms were only identified in patients. Our results support the relevance of gene-gene interactions in myocardial infarction. PMID- 15749686 TI - Importance of troponin T for the risk stratification of normotensive patients with pulmonary embolism. A prospective, cohort study with a three-month follow up. AB - To determine the prognostic importance of troponin T in normotensive patients with pulmonary embolism, we investigated the rate of adverse events in patients with normal and elevated troponin values, during the hospital period and at three months of follow-up. We also calculated the proportion of patients with abnormal troponin values and adverse outcomes who could have been treated with more aggressive therapy according to published criteria. PMID- 15749688 TI - Granulocyte colony-stimulating factor affects serum levels of soluble interleukin 2 receptors after allogeneic stem cell transplantation. AB - Soluble interleukin-2 receptor (sIL-2R) levels were analyzed in 127 stem-cell transplant recipients. Granulocyte-colony stimulating factor (G-CSF) was given to 57 patients after transplantation. We found an association between G-CSF and increased sIL-2R levels. This indicates increased T-cell activation and may be one reason for the previously found increased incidence of acute graft-versus host disease in G-CSF-treated patients. PMID- 15749687 TI - Recipient interferon-gamma 3/3 genotype contributes to the development of chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. AB - Microsatellite polymorphism (CA)n within the first intron of the interferon- gamma gene was assessed in 160 recipients of an allogeneic hematopoietic stem cell transplant (HSCT). IFN- gamma 3/3 was found to be associated with an increased risk of chronic graft-versus-host disease (GvHD) (11/27 vs 26/133, p=0.02). Forward logistic regression analysis confirmed the role of IFN-gamma 3/3 genotype as one of the risk factors for manifestation of chronic GvHD (OR=3.180, p=0.018) together with previous acute GvHD (OR=2.752, p=0.024), cyclosporine A monotherapy (OR=2.607, p=0.029) and malignant disorders (OR=4.371, p=0.032). PMID- 15749689 TI - Newly diagnosed cases of hematologic malignancies in Sardinia in the early 2000: an estimation of their number, age and geographic distribution on the basis of a previous epidemiologic survey. AB - We estimate the number of cases of hematologic malignancies expected to be newly diagnosed in the resident population of Sardinia during the year 2001, and classify the predicted cases according to disease, age and geographic distribution. The implications of these predictions for the Sardinian health care system are discussed, particularly with respect to the development of policies aimed to ensure the most adequate medical care. PMID- 15749690 TI - The brain of LB1, Homo floresiensis. AB - The brain of Homo floresiensis was assessed by comparing a virtual endocast from the type specimen (LB1) with endocasts from great apes, Homo erectus, Homo sapiens, a human pygmy, a human microcephalic, specimen number Sts 5 (Australopithecus africanus), and specimen number WT 17000 (Paranthropus aethiopicus). Morphometric, allometric, and shape data indicate that LB1 is not a microcephalic or pygmy. LB1's brain/body size ratio scales like that of an australopithecine, but its endocast shape resembles that of Homo erectus. LB1 has derived frontal and temporal lobes and a lunate sulcus in a derived position, which are consistent with capabilities for higher cognitive processing. PMID- 15749692 TI - Ca(2+)-induced Ca(2+) release in sensory neurons: low gain amplification confers intrinsic stability. AB - Ca(2+)-induced Ca(2+) release (CICR) is a ubiquitous mechanism by which Ca(2+) release from the endoplasmic reticulum amplifies the trigger Ca(2+) entry and generates propagating Ca(2+) waves. To elucidate the mechanisms that control this positive feedback, we investigated the spatial and temporal kinetics and measured the gain function of CICR in small sensory neurons from mammalian dorsal root ganglions (DRGs). We found that subsurface Ca(2+) release units (CRUs) are under tight local control by Ca(2+) entry, whereas medullar CRUs as a "common pool" system are recruited by inwardly propagating CICR. Active CRUs often displayed repetitive Ca(2+) sparks, conferring the ability to encode a "memory" of neuronal activity well beyond the duration of an action potential. Store Ca(2+) reserve was able to support all CRUs each to fire approximately 15 sparks, excluding use dependent inactivation or store depletion as the major CICR termination mechanism. Importantly, CICR in DRG neurons operated in a low gain, linear regime (gain = 0.54), which conferred intrinsic stability to CICR. Combined with high Ca(2+) current density (-156 pA/pF at -10 mV), such a low gain CICR system generated large intracellular Ca(2+) transients without jeopardizing the stability. These findings provide the first demonstration that CICR operating in a low gain regime can be harnessed to provide a robust and graded amplification of Ca(2+) signal in the absence of counteracting inhibitory mechanism. PMID- 15749693 TI - PSIbase: a database of Protein Structural Interactome map (PSIMAP). AB - Protein Structural Interactome map (PSIMAP) is a global interaction map that describes domain-domain and protein-protein interaction information for known Protein Data Bank structures. It calculates the Euclidean distance to determine interactions between possible pairs of structural domains in proteins. PSIbase is a database and file server for protein structural interaction information calculated by the PSIMAP algorithm. PSIbase also provides an easy-to-use protein domain assignment module, interaction navigation and visual tools. Users can retrieve possible interaction partners of their proteins of interests if a significant homology assignment is made with their query sequences. AVAILABILITY: http://psimap.org and http://psibase.kaist.ac.kr/ PMID- 15749694 TI - REVCOM: a robust Bayesian method for evolutionary rate estimation. AB - MOTIVATION: Evolutionary conservation estimated from a multiple sequence alignment is a powerful indicator of the functional significance of a residue and helps to predict active sites, ligand binding sites, and protein interaction interfaces. Many algorithms that calculate conservation work well, provided an accurate and balanced alignment is used. However, such a strong dependence on the alignment makes the results highly variable. We attempted to improve the conservation prediction algorithm by making it more robust and less sensitive to (1) local alignment errors, (2) overrepresentation of sequences in some branches and (3) occasional presence of unrelated sequences. RESULTS: A novel method is presented for robust constrained Bayesian estimation of evolutionary rates that avoids overfitting independent rates and satisfies the above requirements. The method is evaluated and compared with an entropy-based conservation measure on a set of 1494 protein interfaces. We demonstrated that approximately 62% of the analyzed protein interfaces are more conserved than the remaining surface at the 5% significance level. A consistent method to incorporate alignment reliability is proposed and demonstrated to reduce arbitrary variation of calculated rates upon inclusion of distantly related or unrelated sequences into the alignment. PMID- 15749695 TI - Mutation of active site residues of insulin-degrading enzyme alters allosteric interactions. AB - The active site glutamate (Glu(111)) and the active site histidine (His(112)) of insulin-degrading enzyme (IDE) were mutated. These mutant enzymes exhibit, in addition to a large decrease in catalytic activity, a change in the substrate velocity response from a sigmoidal one seen with the native enzyme (Hill coefficient > 2), to a hyperbolic response. With 2-aminobenzoyl-GGFLRKHGQ-N-(2,4 dinitrophenyl)ethylenediamine as substrate, ATP and triphosphate increase the reaction rate of the wild type enzyme some 50-80-fold. This effect is dampened with glutamate mutants to no effect or less than a 3-fold increase in activity and changed to inhibition with the histidine mutants. Sedimentation equilibrium shows the IDE mutants exhibit a similar oligomeric distribution as the wild type enzyme, being predominantly monomeric, with triphosphate having little if any effect on the oligomeric state. Triphosphate did induce aggregation of many of the IDE mutants. Thus, the oligomeric state of IDE does not correlate with kinetic properties. The His(112) mutants were shown to bind zinc, but with a lower affinity than the wild type enzyme. The glutamate mutants displayed an altered cleavage profile for the peptide beta-endorphin. Wild type IDE cleaved beta-endorphin at Leu(17)-Phe(18) and Phe(18)-Lys(19), whereas the glutamate mutants cleaved at these sites, but in addition at Lys(19)-Asn(20) and at Met(5) Thr(6). Thus, active site mutations of IDE are suggested to not only reduce catalytic activity but also cause local conformational changes that affect the allosteric properties of the enzyme. PMID- 15749696 TI - Increased flexibility as a strategy for cold adaptation: a comparative molecular dynamics study of cold- and warm-active uracil DNA glycosylase. AB - Uracil DNA glycosylase (UDG) is a DNA repair enzyme in the base excision repair pathway and removes uracil from the DNA strand. Atlantic cod UDG (cUDG), which is a cold-adapted enzyme, has been found to be up to 10 times more catalytically active in the temperature range 15-37 degrees C as compared with the warm-active human counterpart. The increased catalytic activity of cold-adapted enzymes as compared with their mesophilic homologues are partly believed to be caused by an increase in the structural flexibility. However, no direct experimental evidence supports the proposal of increased flexibility of cold-adapted enzymes. We have used molecular dynamics simulations to gain insight into the structural flexibility of UDG. The results from these simulations show that an important loop involved in DNA recognition (the Leu(272) loop) is the most flexible part of the cUDG structure and that the human counterpart has much lower flexibility in the Leu(272) loop. The flexibility in this loop correlates well with the experimental k(cat)/K(m) values. Thus, the data presented here add strong support to the idea that flexibility plays a central role in adaptation to cold environments. PMID- 15749697 TI - Drosophila mitochondrial transcription factor B1 modulates mitochondrial translation but not transcription or DNA copy number in Schneider cells. AB - We report the cloning and molecular analysis of Drosophila mitochondrial transcription factor (d-mtTF) B1. An RNA interference (RNAi) construct was designed that reduces expression of d-mtTFB1 to 5% of its normal level in Schneider cells. In striking contrast with our previous study on d-mtTFB2, we found that RNAi knock-down of d-mtTFB1 does not change the abundance of specific mitochondrial RNA transcripts, nor does it affect the copy number of mitochondrial DNA. In a corollary manner, overexpression of d-mtTFB1 did not increase either the abundance of mitochondrial RNA transcripts or mitochondrial DNA copy number. Our data suggest that, unlike d-mtTFB2, d-mtTFB1 does not have a critical role in either transcription or regulation of the copy number of mitochondrial DNA. Instead, because we found that RNAi knockdown of d-mtTFB1 reduces mitochondrial protein synthesis, we propose that it serves its primary role in modulating translation. Our work represents the first study to document the role of mtTFB1 in vivo and establishes clearly functional differences between mtTFB1 and mtTFB2. PMID- 15749698 TI - Nuclear localization and chromatin targets of p21-activated kinase 1. AB - Pak1 (p21-activated kinase 1), a conserved, mammalian signaling kinase, is a downstream effector of small GTPases Rac1 and Cdc42 and of growth factor signaling. Until now, a major focus of study has been on the cytosolic functions of Pak1, where it is an important modulator of cytoskeletal reorganization, consequently playing a major role in cell survival, migration, and invasion. In this report, we demonstrate the nuclear localization of Pak1 upon stimulation by epidermal growth factor. Three nuclear localization signals (NLSs) were identified in the N-terminal domain of Pak1. With mutational analysis, the importance of each NLS was elucidated. Mutation of all three NLSs eliminated the nuclear localization of Pak1. Expression of Pak1 as a fusion protein with Gal4 DNA binding domain and Gal4-luciferase activity showed that Pak1 might increase transcription. To identify the potential targets of nuclear Pak1, we used a Pak1 specific chromatin immunoprecipitation-based screening assay and identified a series of Pak1-interacting target chromatins, including phosphofructokinase muscle isoform (PFK-M) and nuclear factor of activated T-cell (NFAT1) genes. Pak1 associated with the upstream enhancer sequence and promoter of PFK-M and was involved in the stimulation of the PFK-M expression. It also associated with a portion of the NFAT1 gene and its upstream region, leading to the repression of NFAT1 expression. These investigations provide proof-of-principle evidence that Pak1 could influence the expression of its putative chromatin targets in both a positive and a negative manner. Together, for the first time, these findings defined the NLSs of the Pak1, its association with chromatin, and the resulting modulation of transcription, thus opening new avenues to further the search for nuclear Pak1 functions and identify putative Pak1-interacting nuclear proteins. PMID- 15749699 TI - Dimerization of nitric oxide-sensitive guanylyl cyclase requires the alpha 1 N terminus. AB - The enzyme nitric oxide-sensitive guanylyl cyclase is an obligate heterodimer consisting of an alpha and a beta subunit. Whereas the C-terminal parts of the subunits have been shown to be sufficient for catalysis, regulation was assigned to the N termini. The central domains have been postulated to be responsible for the formation of alphabeta heterodimers. Here, we have analyzed dimerization by precipitation of various N- and C-terminally truncated alpha(1) mutants with beta(1) wild type or deletion mutants thereof after coexpression in the baculovirus/Sf9 system. In contrast to the current hypothesis, our analysis revealed that an N-terminal region of the alpha(1) subunit (amino acids 61-128) is mandatory for quantitative dimerization. The central domain (amino acids 367 462) contributes but is not sufficient to mediate robust alphabeta interaction. Wild type-like binding of the identified minimum dimerization region of alpha(1) (amino acids 61-462) requires the N-terminal and central region of beta(1) (amino acids 1-385). Furthermore, we observed an unequal stability of the alpha(1) and beta(1) subunit. Whereas beta(1) forms heme containing homodimers and is stable, alpha(1) appears to be prone to misfolding and degradation when heterodimerization is impaired by deletion of important sequences. PMID- 15749700 TI - DNA polymerase lambda mediates a back-up base excision repair activity in extracts of mouse embryonic fibroblasts. AB - Mammalian DNA polymerase (pol) lambda is a member of the X-family of DNA polymerases and has striking enzymatic and structural similarities to mammalian DNA pol beta. Because pol beta provides two important enzymatic activities for base excision repair (BER), we examined whether pol lambda might also contribute to BER. We used extracts from mouse embryonic fibroblasts representing wild-type and null genotypes for pol beta and pol lambda. In combination with neutralizing antibodies against pol beta and pol lambda, our results show a BER deficiency in the pol lambda -/- cell extract compared with extract from isogenic wild-type cells. In addition, the pol lambda antibody strongly reduced in vitro BER in the pol beta -/- cell extract. These data indicate that pol lambda is able to contribute to BER in mouse fibroblast cell extract. PMID- 15749701 TI - Secretion of cartilage oligomeric matrix protein is affected by the signal peptide. AB - Cartilage oligomeric matrix protein (COMP) is a secreted glycoprotein found in the extracellular matrices of skeletal tissues. Mutations associated with two human skeletal dysplasias, pseudoachondroplasia and multiple epiphyseal dysplasia, disturb COMP secretion leading to intracellular accumulation of mutant COMP, especially in chondrocytes. Here we show that the manifestation of this secretory defect is dramatically influenced by the signal peptide that targets COMP for secretion. The comparison of wild type and mutant COMP secretion directed by the COMP or BM40 signal peptide in HEK-293 cells and rat chondrosarcoma cells revealed that the BM40 signal peptide substantially enhances secretion of mutant COMP that accumulates in endoplasmic reticulum-like structures when targeted by its own signal peptide. Additionally, we demonstrate that mutant COMP forms mixed pentamers with wild type COMP. Our findings suggest that the secretory defect in pseudoachondroplasia and multiple epiphyseal dysplasia is not specific for chondrocytes, nor does it require interaction of mutant COMP with other matrix proteins prior to transport from the cell. They also imply a previously unappreciated role for the signal peptide in the regulation of protein secretion beyond targeting to the endoplasmic reticulum. PMID- 15749702 TI - Internal ribosome entry sites in cellular mRNAs: mystery of their existence. AB - Although studies on viral gene expression were essential for the discovery of internal ribosome entry sites (IRESs), it is becoming increasingly clear that IRES activities are present in a significant number of cellular mRNAs. Remarkably, many of these IRES elements initiate translation of mRNAs encoding proteins that protect cells from stress (when the translation of the vast majority of cellular mRNAs is significantly impaired). The purpose of this review is to summarize the progress on the discovery and function of cellular IRESs. Recent findings on the structures of these IRESs and specifically regulation of their activity during nutritional stress, differentiation, and mitosis will be discussed. PMID- 15749703 TI - Identification and characterization of GIV, a novel Galpha i/s-interacting protein found on COPI, endoplasmic reticulum-Golgi transport vesicles. AB - In this report, we characterize GIV (Galpha-interacting vesicle-associated protein), a novel protein that binds members of the Galpha(i) and Galpha subfamilies of heterotrimeric G proteins. The Galpha(s) interaction site was mapped to an 83-amino acid region of GIV that is enriched in highly charged amino acids. BLAST searches revealed two additional mammalian family members, Daple and an uncharacterized protein, FLJ00354. These family members share the highest homology at the Galpha binding domain, are homologous at the N terminus and central coiled coil domain but diverge at the C terminus. Using affinity-purified IgG made against two different regions of the protein, we localized GIV to COPI, endoplasmic reticulum (ER)-Golgi transport vesicles concentrated in the Golgi region in GH3 pituitary cells and COS7 cells. Identification as COPI vesicles was based on colocalization with beta-COP, a marker for these vesicles. GIV also codistributes in the Golgi region with endogenous calnuc and the KDEL receptor, which are cis Golgi markers and with Galpha(i3)-yellow fluorescent protein expressed in COS7 cells. By immunoelectron microscopy, GIV colocalizes with beta COP and Galpha(i3) on vesicles found in close proximity to ER exit sites and to cis Golgi cisternae. In cell fractions prepared from rat liver, GIV is concentrated in a carrier vesicle fraction (CV2) enriched in ER-Golgi transport vesicles. beta-COP and several Galpha subunits (Galpha(i1-3), Galpha(s)) are also most enriched in CV2. Our results demonstrate the existence of a novel Galpha interacting protein associated with COPI transport vesicles that may play a role in Galpha-mediated effects on vesicle trafficking within the Golgi and/or between the ER and the Golgi. PMID- 15749704 TI - AP2 clathrin adaptor complex, but not AP1, controls the access of the major histocompatibility complex (MHC) class II to endosomes. AB - Newly synthesized MHC II alpha- and beta-chains associated with the invariant chain chaperone (Ii) enter the endocytic pathway for Ii degradation and loading with peptides before transport to the cell surface. It is unclear how alphabetaIi complexes are sorted from the Golgi apparatus and directed to endosomes. However, indirect evidence tends to support direct transport involving the AP1 clathrin adaptor complex. Surprisingly, we show here that knocking down the production of AP1 by RNA interference did not affect the trafficking of alphabetaIi complexes. In contrast, AP2 depletion led to a large increase in surface levels of alphabetaIi complexes, inhibited their rapid internalization, and strongly delayed the appearance of mature MHC II in intracellular compartments. Thus, in the cell systems studied here, rapid internalization of alphabetaIi complexes via an AP2-dependent pathway represents a key step for MHC II delivery to endosomes and lysosomes. PMID- 15749705 TI - Mechanism of human SIRT1 activation by resveratrol. AB - The NAD+-dependent protein deacetylase family, Sir2 (or sirtuins), is important for many cellular processes including gene silencing, regulation of p53, fatty acid metabolism, cell cycle regulation, and life span extension. Resveratrol, a polyphenol found in wines and thought to harbor major health benefits, was reported to be an activator of Sir2 enzymes in vivo and in vitro. In addition, resveratrol was shown to increase life span in three model organisms through a Sir2-dependent pathway. Here, we investigated the molecular basis for Sir2 activation by resveratrol. Among the three enzymes tested (yeast Sir2, human SIRT1, and human SIRT2), only SIRT1 exhibited significant enzyme activation ( approximately 8-fold) using the commercially available Fluor de Lys kit (BioMol). To examine the requirements for resveratrol activation of SIRT1, we synthesized three p53 acetylpeptide substrates either lacking a fluorophore or containing a 7 amino-4-methylcoumarin (p53-AMC) or rhodamine 110 (p53-R110). Although SIRT1 activation was independent of the acetylpeptide sequence, resveratrol activation was completely dependent on the presence of a covalently attached fluorophore. Substrate competition studies indicated that the fluorophore decreased the binding affinity of the peptide, and, in the presence of resveratrol, fluorophore containing substrates bound more tightly to SIRT1. Using available crystal structures, a model of SIRT1 bound to p53-AMC peptide was constructed. Without resveratrol, the coumarin of p53-AMC peptide is solvent-exposed and makes no significant contacts with SIRT1. We propose that binding of resveratrol to SIRT1 promotes a conformational change that better accommodates the attached coumarin group. PMID- 15749706 TI - Asparagine deamidation perturbs antigen presentation on class II major histocompatibility complex molecules. AB - Post-translational protein modifications can be recognized by B and T lymphocytes and can potentially make "self"-proteins appear foreign to the immune system. Such modifications may directly affect major histocompatibility complex restricted T cell recognition of processed peptides or may perturb the processing events that generate such peptides. Using the tetanus toxin C fragment protein as a test case, we show that spontaneous deamidation of asparagine residues interferes with processing by the enzyme asparagine endopeptidase (AEP) and contributes to diminished antigen presentation. Deamidation inhibits AEP action either directly, when asparagine residues targeted by AEP are modified, or indirectly, when adjacent Asn residues are deamidated. Thus, deamidation of long lived self-proteins may qualitatively or quantitatively affect the spectrum of self-peptides displayed to T cells and may thereby contribute to the onset or exacerbation of autoimmune disease. PMID- 15749707 TI - A novel ligand-independent apoptotic pathway induced by scavenger receptor class B, type I and suppressed by endothelial nitric-oxide synthase and high density lipoprotein. AB - Scavenger receptor class B, type I (SR-BI)/ApoE double null mice develop severe atherosclerosis within 4 weeks, whereas ApoE null mice take several months to develop the disease, indicating that SR-BI plays a pivotal role in atherosclerosis. Importantly, SR-BI/ApoE double null mice have lower plasma cholesterol levels than ApoE null mice, suggesting involvement of a non-lipids mechanism. In the present study, we revealed a novel ligand-independent apoptotic pathway induced by SR-BI, and regulated by endothelial nitric-oxide synthase (eNOS) and high density lipoprotein (HDL). SR-BI significantly induces apoptosis in three independent cell systems. In contrast to known ligand-dependent apoptotic pathways, SR-BI-induced apoptosis is ligand-independent. We further showed that SR-BI-induced apoptosis is suppressed by eNOS and HDL. By using a single site mutation, we demonstrated that SR-BI induces apoptosis through a highly conserved CXXS redox motif. We finally demonstrated that SR-BI-induced apoptosis is via the caspase-8 pathway. We hypothesize that in healthy cells, the SR-BI apoptotic pathway is turned off by eNOS and HDL which prevents inappropriate apoptotic damage to the vascular wall. When HDL levels are low, oxidative stress causes the relocation of eNOS away from caveolae, which turns on SR-BI-induced apoptosis and rapidly clears damaged cells to prevent further inflammatory damage to neighboring cells. The current studies offer a new paradigm in which to study the non-cholesterol effects of SR-BI, HDL, and eNOS on the development of atherosclerosis and potentially other cardiovascular diseases. PMID- 15749708 TI - Femtomolar transition state analogue inhibitors of 5'-methylthioadenosine/S adenosylhomocysteine nucleosidase from Escherichia coli. AB - Escherichia coli 5'-methylthioadenosine/S-adenosyl-homocysteine nucleosidase (MTAN) hydrolyzes its substrates to form adenine and 5-methylthioribose (MTR) or S-ribosylhomocysteine (SRH). 5'-Methylthioadenosine (MTA) is a by-product of polyamine synthesis and SRH is a precursor to the biosynthesis of one or more quorum sensing autoinducer molecules. MTAN is therefore involved in quorum sensing, recycling MTA from the polyamine pathway via adenine phosphoribosyltransferase and recycling MTR to methionine. Hydrolysis of MTA by E. coli MTAN involves a highly dissociative transition state with ribooxacarbenium ion character. Iminoribitol mimics of MTA at the transition state of MTAN were synthesized and tested as inhibitors. 5'-Methylthio-Immucillin A (MT-ImmA) is a slow-onset tight-binding inhibitor giving a dissociation constant (K(i)(*)) of 77 pm. Substitution of the methylthio group with a p-Cl phenylthio group gives a more powerful inhibitor with a dissociation constant of 2 pm. DADMe-Immucillins are better inhibitors of E. coli MTAN, since they are more closely related to the highly dissociative nature of the transition state. MT-DADMe-Immucillin-A binds with a K(i)(*) value of 2 pm. Replacing the 5'-methyl group with other hydrophobic groups gave 17 transition state analogue inhibitors with dissociation constants from 10(-12) to 10(-14) m. The most powerful inhibitor was 5'-p-Cl-phenylthio-DADMe-Immucillin-A (pClPhT-DADMe-ImmA) with a K(i)(*) value of 47 fm (47 x 10(-15) m). These are among the most powerful non covalent inhibitors reported for any enzyme, binding 9-91 million times tighter than the MTA and SAH substrates, respectively. The inhibitory potential of these transition state analogue inhibitors supports a transition state structure closely resembling a fully dissociated ribooxacarbenium ion. Powerful inhibitors of MTAN are candidates to disrupt key bacterial pathways including methylation, polyamine synthesis, methionine salvage, and quorum sensing. The accompanying article reports crystal structures of MTAN with these analogues. PMID- 15749709 TI - The low density lipoprotein receptor-related protein (LRP) is a novel beta secretase (BACE1) substrate. AB - BACE is a transmembrane protease with beta-secretase activity that cleaves the amyloid precursor protein (APP). After BACE cleavage, APP becomes a substrate for gamma-secretase, leading to release of amyloid-beta peptide (Abeta), which accumulates in senile plaques in Alzheimer disease. APP and BACE are co internalized from the cell surface to early endosomes. APP is also known to interact at the cell surface and be internalized by the low density lipoprotein receptor-related protein (LRP), a multifunctional endocytic and signaling receptor. Using a new fluorescence resonance energy transfer (FRET)-based assay of protein proximity, fluorescence lifetime imaging (FLIM), and co immunoprecipitation we demonstrate that the light chain of LRP interacts with BACE on the cell surface in association with lipid rafts. Surprisingly, the BACE LRP interaction leads to an increase in LRP C-terminal fragment, release of secreted LRP in the media and subsequent release of the LRP intracellular domain from the membrane. Taken together, these data suggest that there is a close interaction between BACE and LRP on the cell surface, and that LRP is a novel BACE substrate. PMID- 15749710 TI - Interaction between ArgR and AhrC controls regulation of arginine metabolism in Lactococcus lactis. AB - The expression of arginine metabolism in Lactococcus lactis is controlled by the two homologous transcriptional regulators ArgR and AhrC. Genome sequence analyses have shown that the occurrence of multiple homologues of the ArgR family of transcriptional regulators is a common feature of many low-G + C Gram-positive bacteria. Detailed studies of ArgR type regulators have previously only been carried out in bacteria containing single regulators. Here, we present a first characterization of the two L. lactis arginine regulators by means of gel retardation and DNase I footprinting. ArgR of L. lactis was shown to bind to the promoter regions of both the arginine biosynthetic argCJDBF operon and the arginine catabolic arcABD1C1C2TD2yvaD operon, but in an arginine-independent manner. Surprisingly, AhrC alone was unable to bind to DNA. Arginine-dependent DNA binding was obtained by mixing the two regulators in gel retardation assays. With both regulators present, the addition of arginine led to increased binding of ArgR-AhrC to the biosynthetic argC promoter but also to diminished binding to the catabolic arcA promoter. Footprinting showed ArgR-AhrC protection of regions containing ARG box operator sequences preceding argC. In the absence of AhrC, ArgR protected sites in the arcA promoter region with similarity to ARG box half sites, here called ARC boxes. We propose a model for repression of arginine biosynthesis and activation of catabolism by anti-repression, involving arginine dependent interaction between the two L. lactis regulator proteins, ArgR and AhrC. PMID- 15749711 TI - Molecular compatibility of the channel gate and the N terminus of S5 segment for voltage-gated channel activity. AB - Voltage-gated ion channels are modular proteins designed by the structural linkage of a voltage sensor and a pore domain. The functional coupling of these two protein modules is a subject of intense research. A major focus has been directed to decipher the role of the S4-S5 linker and the C-end of the inner pore helix in channel gating. However, the contribution of the cytosolic N terminus of S5 remains elusive. To address this issue, we used a chimeric subunit that linked the voltage sensor of the Shaker channel to the prokaryotic KcsA pore domain (denoted as Shaker-KcsA). This chimera preserved the Shaker sequences at both the N terminus of S5 and the C-end of S6. Chimeric Shaker-KcsA subunits did not form functional homomeric channels but were synthesized, folded, and trafficked to the cell surface, as evidenced by their co-assembly with Shaker wild type subunits. Sequential substitution of Shaker amino acids at the C-end of S6 and the N terminus of S5 by the corresponding KcsA created voltage-sensitive channels with voltage-dependent properties that asymptotically approached those of the wild type Shaker channel. Noteworthy, substitution of the region encompassing Phe(401) Phe(404) at the N-end of Shaker S5 by KcsA residues resulted in a significant gain in voltage sensitivity of the chimeras. Furthermore, analysis of channel function at high [K(+)](o) revealed that the Phe(401)-Phe(404) region is an important molecular determinant for competent coupling of voltage sensing and pore opening. Taken together, these findings indicate that complete replacement of Shaker S5 and S6 by KcsA M1 and M2 is required for voltage-dependent gating of the prokaryotic channel. In addition, our results imply that the region encompassing Phe(401)-Phe(404) in Shaker is involved in protein-protein interactions with the voltage sensor, and signal to the Phe(401) in the S5 segment as a key molecular determinant to pair the voltage sensor and the pore domain. PMID- 15749713 TI - The interaction of LEDGF/p75 with integrase is lentivirus-specific and promotes DNA binding. AB - We have previously shown that the p75 isoform of the transcriptional co-activator lens epithelium-derived growth factor (LEDGF) interacts tightly with human immunodeficiency virus (HIV)-1 integrase (IN) and is essential for nuclear targeting of this protein in human cells (Cherepanov, P., Maertens, G., Proost, P., Devreese, B., Van Beeumen, J., Engelborghs, Y., De Clercq, E., and Debyser, Z. (2003) J. Biol. Chem. 278, 372-381; Maertens, G., Cherepanov, P., Pluymers, W., Busschots, K., De Clercq, E., Debyser, Z., and Engelborghs, Y. (2003) J. Biol. Chem. 278, 33528-33539). Here the interaction between recombinant LEDGF/p75 and HIV-1 IN was examined in a pull-down binding test. LEDGF/p75 was shown to increase the solubility of HIV-1 IN. Next, fluorescent correlation spectroscopy was used to measure the interaction of LEDGF/p75 or the complex of HIV-1 IN and LEDGF/p75 with a specific double-stranded DNA oligonucleotide. Whereas LEDGF/p75 displayed only a moderate affinity for DNA, it strongly promoted the binding of HIV-1 IN to DNA. This effect was specific for the p75 isoform of LEDGF and was not seen with p52. In the pull-down assay LEDGF/p75 interacted with HIV-1, HIV-2, and feline immunodeficiency virus IN, but not with the IN of human T-cell lymphotropic virus type 2, Moloney murine leukemia virus, or Rous sarcoma virus. These results strongly suggest that the interaction of LEDGF/p75 with IN is specific to lentiviridae. LEDGF/p75 stimulated the binding of HIV-1 and HIV-2 IN, but not Moloney murine leukemia virus or Rous sarcoma virus IN, to an aspecific DNA. These results provide supporting evidence for our hypothesis that LEDGF/p75 plays a role in the tethering of lentiviral IN to the chromosomal DNA. PMID- 15749712 TI - Cullins 3a and 3b assemble with members of the broad complex/tramtrack/bric-a brac (BTB) protein family to form essential ubiquitin-protein ligases (E3s) in Arabidopsis. AB - Selective modification of proteins by ubiquitination is directed by diverse families of ubiquitin-protein ligases (or E3s). A large collection of E3s use Cullins (CULs) as scaffolds to form multisubunit E3 complexes in which the CUL binds a target recognition subcomplex and the RBX1 docking protein, which delivers the activated ubiquitin moiety. Arabidopsis and rice contain a large collection of CUL isoforms, indicating that multiple CUL-based E3s exist in plants. Here we show that Arabidopsis CUL3a and CUL3b associate with RBX1 and members of the broad complex/tramtrack/bric-a-brac (BTB) protein family to form BTB E3s. Eighty genes encoding BTB domain-containing proteins were identified in the Arabidopsis genome, indicating that a diverse array of BTB E3s is possible. In addition to the BTB domain, the encoded proteins also contain various other interaction motifs that likely serve as target recognition elements. DNA microarray analyses show that BTB genes are expressed widely in the plant and that tissue-specific and isoform-specific patterns exist. Arabidopsis defective in both CUL3a and CUL3b are embryo-lethal, indicating that BTB E3s are essential for plant development. PMID- 15749714 TI - A single Mms2 "key" residue insertion into a Ubc13 pocket determines the interface specificity of a human Lys63 ubiquitin conjugation complex. AB - Human Ubc13 and Mms2 (or its homolog, Uev1) form a unique ubiquitin-conjugating enzyme (Ubc) complex that generates atypical Lys(63)-linked ubiquitin conjugates. Such conjugates are attached to specific targets that modulate the activity of various cellular processes including DNA repair, mitotic progression, and nuclear factor-kappaB signaling. Whereas Ubc13 is a typical Ubc, Mms2 is a non-catalytic Ubc variant. Substantial biochemical evidence has revealed a mechanism whereby Mms2 properly orients ubiquitin to allow for Lys(63) conjugation by Ubc13; however, how this specific Ubc13-Mms2 complex is formed and why Mms2 does not form a complex with other Ubcs have not been reported. In order to address these questions, we used a structure-based approach to design mutations and characterize the human Ubc13-Mms2 interface. We used the yeast two-hybrid assay, glutathione S-transferase pull-downs, and surface plasmon resonance to test in vivo and in vitro binding. These experiments were paired with functional complementation and ubiquitin conjugation studies to provide in vivo and in vitro functional data. The results in this study allowed us to identify important residues of the Ubc13-Mms2 interface, determine a correlation between heterodimer formation and function, and conclude why Mms2 forms a specific complex with Ubc13 but not other Ubc proteins. PMID- 15749715 TI - The inner mitochondrial membrane has aquaporin-8 water channels and is highly permeable to water. AB - Mitochondria are remarkably plastic organelles constantly changing their shape to fulfil their various functional activities. Although the osmotic movement of water into and out of the mitochondrion is central for its morphology and activity, the molecular mechanisms and the pathways for water transport across the inner mitochondrial membrane (IMM), the main barrier for molecules moving into and out of the organelle, are completely unknown. Here, we show the presence of a member of the aquaporin family of water channels, AQP8, and demonstrate the strikingly high water permeability (Pf) characterizing the rat liver IMM. Immunoblotting, electron microscopy, and biophysical studies show that the largest mitochondria feature the highest AQP8 expression and IMM Pf. AQP8 was also found in the mitochondria of other organs, whereas no other known aquaporins were seen. The osmotic water transport of liver IMM was partially inhibited by the aquaporin blocker Hg2+, while the related activation energy remained low, suggesting the presence of a Hg2+-insensitive facilitated pathway in addition to AQP8. It is suggested that AQP8-mediated water transport may be particularly important for rapid expansions of mitochondrial volume such as those occurring during active oxidative phosphorylation and those following apoptotic signals. PMID- 15749716 TI - Evidence for the involvement of the cannabinoid CB2 receptor and its endogenous ligand 2-arachidonoylglycerol in 12-O-tetradecanoylphorbol-13-acetate-induced acute inflammation in mouse ear. AB - 2-Arachidonoylglycerol is an endogenous ligand for the cannabinoid receptors. Two types of cannabinoid receptors have been identified to date. The CB1 receptor is abundantly expressed in the brain, and assumed to be involved in the attenuation of neurotransmission. On the other hand, the physiological roles of the CB2 receptor, mainly expressed in several types of inflammatory cells and immunocompetent cells, have not yet been fully elucidated. In this study, we investigated possible pathophysiological roles of the CB2 receptor and 2 arachidonoylglycerol in acute inflammation in mouse ear induced by the topical application of 12-O-tetradecanoylphorbol-13-acetate. We found that the amount of 2-arachidonoylglycerol was markedly augmented in inflamed mouse ear. In contrast, the amount of anandamide, another endogenous cannabinoid receptor ligand, did not change markedly. Importantly, 12-O-tetradecanoylphorbol-13-acetate-induced ear swelling was blocked by treatment with SR144528, a CB2 receptor antagonist, suggesting that the CB2 receptor is involved in the swelling. On the other hand, the application of AM251, a CB1 receptor antagonist, exerted only a weak suppressive effect. The application of SR144528 also reduced the 12-O tetradecanoylphorbol-13-acetate-induced production of leukotriene B(4) and the infiltration of neutrophils in the mouse ear. Interestingly, the application of 2 arachidonoylglycerol to the mouse ear evoked swelling, which was abolished by treatment with SR144528. Nitric oxide was suggested to be involved in the ear swelling induced by 2-arachidonoylglycerol. These results suggest that the CB2 receptor and 2-arachidonoylglycerol play crucial stimulative roles during the course of inflammatory reactions. PMID- 15749719 TI - Concerning: 'Why has antibiotic prescribing for respiratory illness declined in primary care? A longitudinal study using the General Practice Research Database'. PMID- 15749717 TI - The keratan sulfate disaccharide Gal(6S03) beta1,4-GlcNAc(6S03) modulates interleukin 12 production by macrophages in murine Thy-1 type autoimmune disease. AB - It has been reported that disaccharides of the glycosaminoglycans (GAGs), heparin, or heparan sulfate suppress the production of cytokines. Therefore, we examined the effects of GAGs (keratan sulfate, hyaluronan, chondroitin, chondroitin sulfate, and heparin sulfate) disaccharides on production of interleukin (IL)-12, a pivotal cytokine in the Th-1 type immune system. Among the GAG disaccharides, only a keratan sulfate disaccharide, Gal(6-SO(3))-GlcNAc(6 SO(3)) (L4), suppressed IL-12 production in macrophages stimulated with lipopolysaccharides and interferon-gamma. Neither keratan sulfate chains nor keratan sulfate tetrasaccharides elicited any change in the IL-12 production. N Acetyl-lactosamine, Gal-GlcNAc (LacNAc), also did not change IL-12 production. These results indicated that a certain size, i.e. disaccharide and sulfate, are essential to suppress IL-12 production. L4 was then applied to MRL-lpr/lpr mice, a Th-1 type autoimmune disease model. The treatment of MRL-lpr/lpr mice with L4 1) decreased in serum IL-12, 2) induced apoptosis in T cells in lymph nodes thereby suppressing lymphoaccumulation, and 3) suppressed hypergammaglobulinemia and glomerulonephritis. We showed previously that IL-12 suppresses cell death of T cells, thereby enhancing the lymphoaccumulation in MRL-lpr/lpr mice. Moreover, it has been reported that IL-12 deficiency in MRL-lpr/lpr mice diminishes lymphoaccumulation and delays glomerulonephritis. The treatment with L4 suppressed phosphoprotein kinase C and phosphoinositide 3-kinase expression in macrophages, suggesting that L4 suppresses IL-12 production by inhibiting phosphoprotein kinase C and phosphoinositide 3-kinase pathways. PMID- 15749720 TI - Evaluation of an independent, radiographer-led community diagnostic ultrasound service provided to general practitioners. AB - BACKGROUND: Health care services traditionally offered in a secondary setting are increasingly being offered in a primary setting. There has been little assessment of quality and efficiency of diagnostic services such as ultrasound delivered in primary settings and no studies have looked at independently provided services. AIMS: To assess the benefits and disadvantages of a radiographer delivered, primary care-based mobile diagnostic ultrasound service by comparing it to an NHS Trust diagnostic ultrasound service. DESIGN: A retrospective, comparative study. SETTING: A primary care area in the West Midlands. METHOD: Random samples of 200 and 193 adult patients who underwent diagnostic ultrasound in 2001/2002 with the community and NHS Trust services respectively, and all GP principals in the area were identified. Patient access (including wait for appointments), patient and GP satisfaction, clinical quality of services, and cost-effectiveness were assessed by postal questionnaires, interviews, review of stored ultrasound images, patient record review and collection of data on unit costs. RESULTS: Mean wait for an appointment was 17.44 (15.85-19.02) and 44.53 days (38.83-50.23) for the community and NHS Trust services respectively. Response rates from the community and hospital patient groups were 52.9 percent and 44.6 percent, respectively. Demographic characteristics of the two groups of respondents did not differ significantly, therefore justifying comparison between the two groups of respondents. High proportions of patients from both services reported time and location of appointment as convenient. Access to secondary care following an abnormal ultrasound was not systematically different for the services. Patients were highly satisfied with both services. GPs were markedly less satisfied with the NHS Trust service compared to the community service. Quality of stored ultrasound images and reports were comparable for the services. Cost per abnormality detected was higher for the community service (107.69 pound sterling compared to 77.35 pound sterling for the NHS Trust service, not statistically significant). CONCLUSION: The community diagnostic ultrasound service offers reduced waiting times compared to the NHS Trust service, and is of comparable quality. This benefit, together with high patient and GP satisfaction levels, may justify the possible reduced cost-effectiveness of the service compared to the NHS Trust service. PMID- 15749721 TI - Who has time to sleep? AB - BACKGROUND: Both marriage and education appear to confer a protective effect on health. Few studies have examined the extent to which both undersleeping and oversleeping explain these relationships. I examined whether marital status, educational background, and other sociodemographic variables are associated with higher-risk sleep durations. METHODS: Over 7000 individuals aged 25-64 collected time-use diary data during a 24-h period. For both weekdays and weekend days, separate multinomial logistic regression models were estimated using three categories of sleep duration as the outcome category. RESULTS: Unmarried individuals are significantly more likely to sleep a short amount on both weekdays and weekends compared to married people. Single people also are significantly more likely to sleep a long amount. People with less than a college education are significantly more likely to sleep both a short amount and a long amount on weekdays relative to the college educated. CONCLUSIONS: High-risk sleep durations (short sleeping and long sleeping) are positively associated with sociodemographic categories associated with poorer health. More research should investigate whether social inequalities in health can be explained in part by variation in sleep duration. PMID- 15749722 TI - Breast cancer services--a population-based study of service reorganization. AB - BACKGROUND: The Calman-Hine Report in England and Wales and the Campbell Report in Northern Ireland initiated a major reorganization of cancer services with the aim of improving the provision and delivery of care to cancer patients. In this paper, breast cancer patients diagnosed in 1996 and those diagnosed in 2001 are compared in order to examine the extent of service change in Northern Ireland. METHODS: Patients living in Northern Ireland with invasive breast cancer diagnosed in either 1996 or 2001 were identified from the Northern Ireland Cancer Registry database. Information relating to the delivery of cancer care to these patients was collected by retrospective review of their case notes. RESULTS: Breast cancer surgery was performed in fewer hospitals (13 versus 21) by fewer surgeons (19 versus 40) with 98 percent of patients operated on by designated breast surgeons in 2001. Clinically relevant axillary node excision increased with 765 (87 percent) patients having six or more nodes excised in 2001 compared to 515 (67 percent) in 1996 (p < 0.001). Recording of oestrogen receptor (ER) status improved from 23 percent in 1996 to 91 percent in 2001 (p < 0.001) and 81 percent of patients received hormone therapy appropriate to their ER status in 2001 compared with 6 percent in 1996 (p < 0.001). Communication between hospitals and patients and their GPs also improved. CONCLUSION: A significant change in service provision and delivery has occurred in Northern Ireland in line with the recommendations of the Campbell report. Further work is underway to ascertain the extent to which these changes may have impacted on patient outcomes. PMID- 15749723 TI - Relationship between travel time to the nearest hospital and survival from ruptured abdominal aortic aneurysms: record linkage study. AB - BACKGROUND: There is currently a trend in Britain to concentrate specialist services in a smaller number of hospitals in order to improve outcomes. However, for rapidly life-threatening conditions such as ruptured abdominal aortic aneurysms (RAAAs), the resulting increased travel time to hospital might adversely affect survival. This study aimed to evaluate the relationship between travel time to the nearest hospital and survival from RAAAs in West Sussex. METHODS: Information was collected regarding outcome, postcode, age, sex and diagnoses for all West Sussex residents who had a RAAA between January 1996 and September 1999, including admissions and deaths, wherever they occurred. Deprivation scores were calculated based on postcode. Potential travel time to the nearest hospital was calculated using "Microsoft AutoRoute Express" and its effect on outcome was analysed using multiple logistic regression. RESULTS: After adjusting for age, sex, Townsend deprivation score and nearest hospital, the odds ratio for survival associated with a 10 min increase in potential travel time to the nearest hospital was 0.97 (0.88 unadjusted) (95 percent confidence interval for adjusted odds ratio: 0.70 to 1.34; p = 0.86). CONCLUSION: Several previous studies suggested better survival with longer distances travelled. They are likely to be biased by omission of community deaths. This methodologically better study found no such relationship between RAAA survival and travelling time to hospital in West Sussex, although confidence intervals were wide. This is particularly pertinent while there is a push to centralize vascular and other services in the United Kingdom. PMID- 15749724 TI - Professionalism in the 21st century. PMID- 15749726 TI - Geographical distribution of road traffic deaths in England and Wales: place of accident compared with place of residence. AB - BACKGROUND: There are two sources of information on road traffic mortality in England and Wales: ONS records giving the place of residence and police records giving the place of the accident. Use of the police records has been limited by the lack of an obvious denominator to control for population at risk. This study compares the two measures and explores the implications of using the same population denominator for both. METHODS: The number of road traffic deaths occurring in 403 local authority districts in England and Wales during 1995-1999 was compared to the number of deaths to residents in the same period. Both numbers were related to the expected number of deaths to residents and selected environmental risk factors using regression techniques. RESULTS: Large differences were found between the number of deaths in each district and the number of deaths to residents. The expected number of deaths to residents was the strongest predictor of both observed totals. The number of deaths in the district and the number divided by expected deaths of residents were highly predictable from road accident risk factors, but the number of deaths to residents and the conventional SMR were not. CONCLUSION: Information on the place of residence of road traffic accident fatalities does not show true variations in accident risk. Police records are better for this purpose. The expected number of deaths to residents estimate provides a partial but effective and unbiased control for population effects. PMID- 15749725 TI - Predicted and observed cardiovascular disease in South Asians: application of FINRISK, Framingham and SCORE models to Newcastle Heart Project data. AB - BACKGROUND: South Asian populations in the United Kingdom have a high risk of cardiovascular disease (CVD) mortality. Risk prediction models appear to be inaccurate in South Asians. OBJECTIVE: To explore the predictive capacity of the FINRISK, Framingham (1991) and SCORE risk prediction models in the Newcastle Heart Project population (n = 1301). METHODS: Mortality data for England and Wales were used to define the expected ranking of CVD risk by country of birth. CVD mortality in the Newcastle Heart Project sample was examined. Risk factor measures were obtained from the Newcastle Heart Project, where 90 percent of South Asians were born in the Indian Subcontinent. The predicted outcomes for FINRISK were acute myocardial infarction and CHD mortality, for Framingham CHD mortality, myocardial infarction, new angina and coronary insufficiency and for SCORE CHD and non-CHD CVD mortality. RESULTS: The FINRISK model predicted in South Asian men combined, compared with Europeans, a risk ratio of 122 per cent (SMR 142) with substantial subgroup heterogeneity, e.g. 154 per cent in Bangladeshis (SMR 151), 129 per cent in Pakistanis (SMR 148), 99 per cent in Indians (SMR 142). The FINRISK risk ratios for South Asian women combined were 160 per cent (SMR 145), for Bangladeshis 184 per cent (SMR 91), Pakistanis 172 per cent (SMR 111) and for Indians 145 per cent (SMR 158). The Framingham model results were very similar to FINRISK, but the SCORE model showed comparatively low 10 year risk in all South Asian groups. Both the Framingham stroke model and the SCORE non-CHD CVD model predicted comparatively low rates, while national data showed these to be high. Control of the five major risk factors was modelled by FINRISK to reduce risk by about 59 per cent in South Asian men and 67 per cent in South Asian women, with some subgroup heterogeneity, compared to 50 per cent in European men and 48 per cent in European women. The Framingham model results were similar. The absolute rates for each ethnic group varied by model. CONCLUSION: The Framingham and FINRISK models gave similar results, mostly following expected patterns, but the SCORE model did not, probably reflecting its lack of inclusion of HDL and diabetes as risk factors. National mortality data and modelled predictions agreed reasonably well for South Asians combined, and Bangladeshi and Pakistani men, but not for Indian men and Pakistani and Bangladeshi women. The varying rates show the limits of modelling. The models suggest the potential gains from controlling major established risk factors could be substantial in South Asians and greater than in Europeans. PMID- 15749727 TI - Communicable disease and health protection quarterly review: July to September 2004 from the Health Protection Agency, Communicable Disease Surveillance Centre. PMID- 15749728 TI - Treatment exhaustion of highly active antiretroviral therapy (HAART) among individuals infected with HIV in the United Kingdom: multicentre cohort study. AB - OBJECTIVES: To investigate whether there is evidence that an increasing proportion of HIV infected patients is starting to experience increases in viral load and decreases in CD4 cell count that are consistent with exhaustion of available treatment options. DESIGN: Multicentre cohort study. SETTING: Six large HIV treatment centres in southeast England. PARTICIPANTS: All individuals seen for care between 1 January 1996 and 31 December 2002. MAIN OUTCOME MEASURES: Exposure to individual antiretroviral drugs and drug classes, CD4 count, plasma HIV RNA burden. RESULTS: Information is available on 16,593 individuals (13,378 (80.6%) male patients, 10,340 (62.3%) infected via homosexual or bisexual sex, 4426 (26.7%) infected via heterosexual sex, median age 34 years). Overall, 10,207 of the 16,593 patients (61.5%) have been exposed to any antiretroviral therapy. This proportion increased from 41.2% of patients under follow up at the end of 1996 to 71.3% of those under follow up in 2002. The median CD4 count and HIV RNA burden of patients under follow up in each year changed from 270 cells/mm3 and 4.34 log10 copies/ml in 1996 to 408 cells/mm3 and 1.89 log10 copies/ml, respectively, in 2002. By 2002, 3060 (38%) of patients who had ever been treated with antiretroviral therapy had experienced all three main classes. Of these, around one quarter had evidence of "viral load failure" with all these three classes. Patients with three class failure were more likely to have an HIV RNA burden > 2.7 log10 copies/ml and a CD4 count < 200 cells/mm3. CONCLUSIONS: The proportion of individuals with HIV infection in the United Kingdom who have been treated has increased gradually over time. A substantial proportion of these patients seem to be in danger of exhausting their options for antiretroviral treatment. New drugs with low toxicity, which are not associated with cross resistance to existing drugs, are urgently needed for such patients. PMID- 15749729 TI - Role of reactive oxygen species and p38 MAPK in the induction of the pro-adhesive endothelial state mediated by IgG from patients with anti-phospholipid syndrome. AB - The association of the presence of anti-phospholipid antibodies (aPL) with thrombosis characterizes the anti-phospholipid syndrome (APS). The activation of the endothelium is a key event in the establishment of the thrombophilic state. However, the intracellular mechanisms leading to endothelial dysfunction are not fully elucidated. We investigated the role of reactive oxygen species (ROS) in the pro-adhesive state elicited by aPL and studied ROS-dependent downstream signaling pathways. Independent incubation of human umbilical vein endothelial cells (HUVEC) with IgG (IgG-APS) from 12 APS patients caused a large and sustained increase in ROS, which was prevented by the antioxidants vitamin C and N-acetyl-L-cysteine. ROS inhibition observed in the presence of diphenylene iodonium and rotenone indicated an involvement of a membrane-bound oxidase and the mitochondrial transport chain as sources of ROS. ROS acted as a second messenger by activating the p38 mitogen-activated protein kinase and its subsequent target, the stress-related transcription factor activating transcription factor-2 (ATF-2). ROS controlled the up-regulation of vascular cell adhesion molecule-1 expression by IgG-APS-stimulated HUVEC and the increase in THP-1 monocytic cells adhesion. The IgG-APS-mediated oxidative stress was observed irrespective of the clinical and biological criterions of the patients studied here. Taken together, these data indicate that the oxidative stress induced by IgG-APS is a key intracellular event that might contribute to the thrombotic complications of APS by controlling the endothelial adhesive phenotype. PMID- 15749730 TI - IFN{alpha} enhances human B-cell chemotaxis by modulating ligand-induced chemokine receptor signaling and internalization. AB - In this study, we show that IFNalpha increases the chemotaxis of human B cells to CCL20, CCL21 and CXCL12 in a dose- and time-dependent manner. The effect was maximal with 2000 IU ml(-1) IFNalpha. It peaked at 24 h and decreased thereafter. At 24 h, IFNalpha had increased B-cell chemotaxis to CCL20 by 20 +/- 6.2% (n = 9, P < 0.002), to CCL21 by 20 +/- 8.5% (n = 14, P < 0.0001) and to CXCL12 by 16.3 +/ 4.2% (n = 12, P < 0.003) without changing CCR6, CCR7 or CXCR4 expression. IFNalpha enhanced the migration of memory B cells to CCL20, CCL21 and CXCL12 2.6 fold more strongly than that of naive B cells. The triggering of chemokine receptors by their ligands resulted in the activation of phosphatidylinositide-3 kinase (PI3K)/protein kinase B (PKB), inhibitory NF-kappaB (IkappaBalpha) RhoA and extracellular signal-regulated protein kinase 1/2 (ERK1/2). All these effectors except ERK1/2 are crucial for B-cell chemotaxis. IFNalpha modulated the requirements for B-cell chemotaxis, which became dependent on ERK1/2, more dependent on PI3K, RhoA and nuclear factor-kappaB but less dependent on Gbetagamma and phospholipase C activation. IFNalpha also decreased ligand-induced chemokine receptor internalization in a manner dependent on PI3K/AKT and RhoA but not on IkappaBalpha and ERK1/2. Our data characterize chemokine receptor signaling in human B cells and clarify the relevance of downstream pathways in B cell chemotaxis and chemokine receptor internalization. They also suggest that non-class I PI3K are involved in B-cell chemotaxis. PMID- 15749731 TI - CD44 cross-linking induces protein kinase C-regulated migration of human T lymphocytes. AB - The cell surface receptor CD44 is widely implicated in leukocyte migration to inflammatory sites. In this study, the responses of human T cells following cross linking of CD44 were examined. We demonstrate that engagement of CD44 using immobilized mAbs or hyaluronan-enriched extracellular matrix lattices induces active migration in T lymphocytes accompanied by cycles of cytoskeletal rearrangement and cell polarization. We have investigated the functional impact and subcellular localization of protein kinase C (PKC) isoenzymes, beta and delta, previously shown by our group to be involved in active T cell locomotion induced by leukocyte function-associated antigen-1 (LFA-1) integrin receptors. PKCbeta was associated with the centrosome and the microtubule-rich tail of the polarized cell and PKCdelta was predominantly located about the region of the microtubule organizing center. A selective pharmacological inhibitor of classical PKC isoforms, Go6976, suppressed lymphocyte polarization and migration following CD44 ligation. Selective targeting of PKCdelta using the pharmacological inhibitor rottlerin or a pseudosubstrate-blocking peptide reduced CD44-activated cell migration but did not completely ablate it. Our data demonstrate that ligation of CD44 induces phenotypic changes, cytoskeletal rearrangements and redistribution of PKC isoforms beta and delta, resulting in cell migration, as previously described for the cell surface receptor, LFA-1. This suggests potential convergence of intracellular signaling pathways induced via CD44 and LFA-1 integrin. PMID- 15749732 TI - Expression of membrane-bound IL-15 by bone marrow fibroblast-like stromal cells in aplastic anemia. AB - In order to explore the relationship between IL-15 and aplastic anemia (AA), bone marrow (BM) fibroblast-like stromal cells (BMFSCs) were obtained from BM samples of 23 AA patients by density centrifugation and primary culturing in vitro. Indirect immunofluorescence labeling as well as flow cytometry and confocal laser scanning microscopy analysis were used to determine the expression of membrane bound IL-15 (mIL-15) on the surface of BMFSCs derived from AA patients (AA BMFSCs). The effects of IFN-gamma and cyclosporin A (CsA) on the expression of mIL-15 were also investigated. [(3)H]thymidine incorporation test as well as specific antibody inhibition and Transwell separation experiment was adopted to functionally evaluate the expression of mIL-15 on the surface of AA-BMFSCs. mIL 15 was found to be over-expressed on the surface of AA-BMFSCs. IFN-gamma further significantly up-regulated its expression, which, however, was inhibited by CsA. Interestingly, a tight correlation was found between the expression of mIL-15 and IL-15Ralpha on the surface of AA-BMFSCs. AA-BMFSCs had the capability to stimulate the proliferation of T lymphocytes, which was partly or completely inhibited by using neutralizing anti-IL-15Ralpha antibody, neutralizing anti-IL 15 antibody, blocking anti-IL-2/15Rgamma(c) mAb or Transwell chambers with a 0.3 mum pore size membrane to block the direct cell-to-cell contact between AA-BMFSCs and T cells. Apparently, BMFSCs as the most important component of BM hematopoietic microenvironment usually over-express mIL-15 in AA patients. Therefore, AA-BMFSCs may indirectly participate in the T cell-mediated destruction of hematopoietic progenitors in AA by recruiting T cells to BM and stimulating them in situ. PMID- 15749733 TI - Relationship between intraoperative transoesophageal echocardiography findings and perfusion lung scintigraphy results on first postoperative day. AB - BACKGROUND: Although intraoperative transoesophageal echocardiography (TOE) has been used to detect the occurrence of echogenic macro- and/or microembolic phenomena during total hip arthroplasty (THA), no direct correlation between macroembolism and the formation of pulmonary embolism (PE) has been conclusively determined in early postoperative periods after THA. METHODS: Sixty-two patients scheduled for primary THA were enrolled in this study. Intraoperative TOE images were continuously recorded on videotape and the echogenic events were evaluated throughout surgery. Perfusion lung scintigraphy was performed on the first postoperative day (POD1). RESULTS: Perfusion lung scintigraphy revealed the existence of PE in nine (15%) of the 62 patients who underwent THA: five (25%) of 20 patients with cemented THA and four (10%) of 42 patients with non-cemented THA. The grading score of intraoperative TOE findings, including the amount of echogenic particles in right atrium, the longest time of echogenesis and the diameter of the largest echogenic particles, did not differ between the groups with and without PE. The sensitivity, specificity, and positive and negative predictive values for the detection of echogenic macroemboli for the prediction of the development of PE on POD1 were 0.78, 0.60, 0.25 and 0.94, respectively. CONCLUSION: Intraoperative TOE monitoring did not predict the occurrence of PE on POD1. PMID- 15749734 TI - Efficacy of A-line AEP Monitor as a tool for predicting acceptable tracheal intubation conditions during sevoflurane anaesthesia. AB - BACKGROUND: It is essential for the clinical anaesthetist to know whether patients are sufficiently anaesthetized to tolerate direct laryngoscopy and endotracheal intubation. Because of the lack of an accurate objective method to determine the level of general anaesthesia, under- or overdosing of anaesthetics may occur. Auditory evoked potential (AEP) is one of several physiological parameters under investigation. We aimed to determine the clinically required depth of anaesthesia, measured by the A-line AEP Monitor and expressed as A-Line ARX Index (AAI) for 90% probability of acceptable conditions for endotracheal intubation. METHODS: We studied 108 patients anaesthetized by mask with increasing concentration of sevoflurane in 30% oxygen and 70% nitrous oxide. Fentanyl 1.5 microg kg(-1) and glycopyrrolate 0.2 mg were administered intravenously immediately before starting induction of anaesthesia. The monitor was programmed to give an alarm at AAI 10, 15, 20, 25 or 30 according to randomization. When the alarm sounded, the end-expiratory sevoflurane concentration was registered and endotracheal intubation was attempted. Intubation conditions were assessed by an observer blinded to the AAI. RESULTS: At AAI 10 we found acceptable conditions in 91% (confidence interval [CI 72-99%]) of patients. The prediction probability value P(K) of AAI was 0.69 (CI 0.59-0.79) and the P(K) of end-expiratory sevoflurane concentration was 0.93 (CI 0.87-0.99). ED(90) (the AAI with a 90% probability of acceptable intubation conditions) was calculated as 8.5 (CI 0-17.5). CONCLUSIONS: AAI indicates the depth of anaesthesia necessary for acceptable endotracheal intubation conditions. Under the conditions of the present study, end-expiratory sevoflurane concentration was a better predictor and may turn out to be more useful in the clinical setting. PMID- 15749736 TI - Transfusion for a patient of cis-AB blood type undergoing a redo cardiac operation. AB - cis-AB, a rare ABO genotype, is the result of a mutated gene resulting in dual specific hybrid enzymes. A single-point mutation reverses the specificity of human blood group B synthesizing galactosyltransferase. This may lead to misclassification in ABO grouping and adverse transfusion reactions. Recently, the authors experienced a case of a patient with cis-AB blood type undergoing pulmonary valve replacement and tricuspid valvuloplasty. We transfused the patient with Rh+ A packed red blood cell, fresh frozen plasma and platelet concentrates without any clinically significant transfusion reactions. PMID- 15749735 TI - Magnesium sulphate only slightly reduces the shivering threshold in humans. AB - BACKGROUND: Hypothermia may be an effective treatment for stroke or acute myocardial infarction; however, it provokes vigorous shivering, which causes potentially dangerous haemodynamic responses and prevents further hypothermia. Magnesium is an attractive anti-shivering agent because it is used for treatment of postoperative shivering and provides protection against ischaemic injury in animal models. We tested the hypothesis that magnesium reduces the threshold (triggering core temperature) and gain of shivering without substantial sedation or muscle weakness. METHODS: We studied nine healthy male volunteers (18-40 yr) on two randomly assigned treatment days: (1) control and (2) magnesium (80 mg kg( 1) followed by infusion at 2 g h(-1)). Lactated Ringer's solution (4 degrees C) was infused via a central venous catheter over a period of approximately 2 h to decrease tympanic membrane temperature by approximately 1.5 degrees C h(-1). A significant and persistent increase in oxygen consumption identified the threshold. The gain of shivering was determined by the slope of oxygen consumption vs core temperature regression. Sedation was evaluated using a verbal rating score (VRS) from 0 to 10 and bispectral index (BIS) of the EEG. Peripheral muscle strength was evaluated using dynamometry and spirometry. Data were analysed using repeated measures anova; P<0.05 was statistically significant. RESULTS: Magnesium reduced the shivering threshold (36.3 [SD 0.4] degrees C vs 36.6 [0.3] degrees C, P = 0.040). It did not affect the gain of shivering (control, 437 [289] ml min(-1) degrees C(-1); magnesium, 573 [370] ml min(-1) degrees C(-1); P=0.344). The magnesium bolus did not produce significant sedation or appreciably reduce muscle strength. CONCLUSIONS: Magnesium significantly reduced the shivering threshold. However, in view of the modest absolute reduction, this finding is considered to be clinically unimportant for induction of therapeutic hypothermia. PMID- 15749737 TI - Apical location of ferroportin 1 in airway epithelia and its role in iron detoxification in the lung. AB - Ferroportin 1 (FPN1; aka MTP1, IREG1, and SLC40A1), which was originally identified as a basolateral iron transporter crucial for nutritional iron absorption in the intestine, is expressed in airway epithelia and upregulated when these cells are exposed to iron. Using immunofluorescence labeling and confocal microscopic imaging techniques, we demonstrate that in human and rodent lungs, FPN1 localizes subcellularly to the apical but not basolateral membrane of the airway epithelial cells. The role of airway epithelial cells in iron mobilization in the lung was studied in an in vitro model of the polarized airway epithelium. Normal human bronchial epithelial cells, grown on membrane supports until differentiated, were exposed to iron, and the efficiency and direction of iron transportation were studied. We found that these cells can efficiently take up iron across the apical but not basolateral surface in a concentration dependent manner. Most of the iron taken up by the cells is then released into the medium within 8 h in the form of less reactive protein-bound complexes including ferritin and transferrin. Interestingly, iron release also occurred across the apical but not basolateral membrane. Our findings indicate that FPN1, depending on its subcellular location, could have distinct functions in iron homeostasis in different cells and tissues. Although it is responsible for exporting nutrient iron from enterocytes to the circulation in the intestine, it could play a role in iron detoxification in airway epithelial cells in the lung. PMID- 15749738 TI - Amino-terminal TACE prodomain attenuates TNFR2 cleavage independently of the cysteine switch. AB - TNF-alpha-converting enzyme (TACE, ADAM17) cleaves membrane-associated cytokines and receptors and thereby regulates inflammatory and immune events, as well as lung development and mucin production. For example, the TACE-mediated cleavage of the type II 75-kDa TNF receptor (TNFR2) generates a soluble TNF-binding protein that modulates TNF bioactivity. TACE is synthesized as a latent proenzyme that is retained in an inactive state via an interaction between its prodomain and catalytic domain. Although the formation of an intramolecular bond between a cysteine in the prodomain and a zinc atom in the catalytic site had been thought to mediate this inhibitory activity, it was recently reported that the cysteine switch motif is not required. Here, we hypothesized that the amino terminus of the TACE prodomain might contribute to the ability of the prodomain to maintain TACE in an inactive state independently of a cysteine-switch mechanism. We synthesized a 37-amino acid peptide corresponding to TACE amino acids 18-54 (N TACE(18-54)) and assessed whether it possessed TACE inhibitory activity. In an in vitro model assay system, N-TACE(18-54) attenuated TACE-catalyzed cleavage of a TNFR2:Fc substrate. Furthermore, N-TACE(18-54) inhibited constitutive TNFR2 shedding from a human monocytic cell line by 42%. A 19-amino acid, leucine-rich domain, corresponding to TACE amino acids 30-48, demonstrated partial inhibitory activity. In summary, we have identified a subdomain within the amino terminus of the TACE prodomain that attenuates TACE catalytic activity independently of a cysteine-switch mechanism, which provides new insight into the regulation of TACE enzymatic activity. PMID- 15749739 TI - TNF, IFN-gamma, and endotoxin increase expression of DMT1 in bronchial epithelial cells. AB - Regulation of the metal transport protein divalent metal transporter-1 (DMT1) may contribute to the uptake and detoxification of iron by cells resident in the respiratory tract. Inflammation has been associated with an increased availability of this metal resulting in an oxidative stress. Because proinflammatory cytokines and LPS have been demonstrated to affect an elevated expression of DMT1 in a macrophage cell line, we tested the hypothesis that tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and LPS increase DMT1 expression in airway epithelial cells. We used RT-PCR to detect mRNA for both IRE DMT1 and +IRE DMT1 in BEAS-2B cells. Treatment with TNF-alpha, IFN-gamma, or LPS increased both forms. Western blot analysis also demonstrated an increase in the expression of both isoforms of DMT1 after these treatments. Twenty-four hours after exposure of an animal model to TNF-alpha, IFN-gamma, or LPS, a significant increase in pulmonary expression of -IRE DMT1 was seen by immunohistochemistry; the level of +IRE DMT1 was too low in the lung to be visualized using this methodology. Finally, iron transport into BEAS-2B cells was increased after inclusion of TNF-alpha, IFN-gamma, or LPS in the media. We conclude that proinflammatory cytokines and LPS increase mRNA and protein expression of DMT1 in airway cells in vitro and in vivo. Furthermore, both -IRE and +IRE isoforms are elevated after exposures. Increased expression of this protein appears to be included in a coordinated response of the cell and tissue where the function might be to diminish availability of metal. PMID- 15749740 TI - Parathyroid hormone-related protein-mediated responses in pulmonary arteries and veins of newborn lambs. AB - PTHrP has important roles in lung development and function. Here we determined the vasomotor responses of isolated pulmonary arteries and veins of newborn and adult sheep to PTHrP. In vessels constricted with endothelin-1, PTHrP (PTHrP 1 34) caused greater relaxation of veins than of arteries. In both vessel types, relaxation to the peptide was less in adult than in newborn vessels. In newborn lambs, PTHrP-induced relaxation was not affected by endothelium removal, inhibition of eNOS, or inhibition of adenylyl cyclases by SQ-22536. However, relaxation was attenuated by 4-aminopyridine, inhibitor of voltage-dependent potassium channels, in both arteries and veins, and by charybdotoxin, inhibitor of calcium-activated potassium channels, in veins. When vessels were saturated with 8-BrcAMP (3 x 10(-4) M), to eliminate relaxation mediated by endogenous cAMP, PTHrP-induced relaxation was partially attenuated. In vessels treated with 8-BrcAMP (3 x 10(-4) M), 4-aminopyridine but not charybdotoxin inhibited relaxation induced by PTHrP 1-34 in both arteries and veins. Radioimmunoassay showed that, in the presence of a general phosphodiesterase inhibitor, PTHrP caused a concentration-dependent increase in intracellular cAMP content in arteries and veins, which was largely abolished by SQ-22536. Our results demonstrate that PTHrP is a potent vasodilator of pulmonary vessels, with a greater effect in veins than in arteries. Relaxation induced by the peptide contains both cAMP-dependent and -independent components. In both arteries and veins, voltage-dependent potassium channels mediate the response to PTHrP, at least in part, in a cAMP-independent fashion; and in veins, calcium-activated potassium channels may be stimulated by elevated cAMP levels. PMID- 15749741 TI - Exposure to TARC alters beta2-adrenergic receptor signaling in human peripheral blood T lymphocytes. AB - The beta(2)-adrenergic receptor (beta(2)-AR) negatively regulates T cell activity through the activation of the G(s)/adenylyl cyclase/cAMP pathway. beta(2)-AR desensitization, which can be induced by its phosphorylation, may have important consequences for the regulation of T cell function in asthma. In the present study we demonstrate that the C-C chemokine thymus and activation-regulated chemokine (TARC) impairs the ability of beta(2)-agonist fenoterol to activate the cAMP downstream effector cAMP-responsive element binding protein (CREB) in freshly isolated human T cells. The TARC-induced activation of Src kinases resulted in membrane translocation of both G protein-coupled receptor kinase (GRK) 2 and beta-arrestin. Moreover, TARC was able to induce Src-dependent serine phosphorylation of the beta(2)-AR as well as its association with GRK2 and beta arrestin. Finally, in contrast to CREB, phosphorylation of Src and extracellular signal-regulated kinase was enhanced by fenoterol upon TARC pretreatment. In summary, we show for the first time that TARC exposure impairs beta(2)-AR function in T cells. Our data suggest that this is mediated by Src-dependent activation of GRK2, resulting in receptor phosphorylation, binding to beta arrestin, and a switch from cAMP-dependent signaling to activation of the MAPK pathway. We propose that aberrant T cell control in the presence of endogenous beta-agonists promotes T cell-mediated inflammation in asthma. PMID- 15749742 TI - Diesel exhaust activates redox-sensitive transcription factors and kinases in human airways. AB - Diesel exhaust (DE) is a major component of airborne particulate matter. In previous studies we have described the acute inflammatory response of the human airway to inhaled DE. This was characterized by neutrophil, mast cell, and lymphocyte infiltration into the bronchial mucosa with enhanced epithelial expression of IL-8, Gro-alpha, and IL-13. In the present study, we investigated whether redox-sensitive transcription factors were activated as a consequence of DE exposure, consistent with oxidative stress triggering airway inflammation. In archived biopsies from 15 healthy subjects exposed to DE [particulates with a mass median diameter of <10 mum, 300 microg/m3] and air, immunohistochemical staining was used to quantify the expression of the transcription factors NF kappaB (p65) and AP-1 (c-jun and c-fos), as well their upstream MAPKs, p38 and JNK, in the bronchial epithelium. In addition, phosphorylation of tyrosine residues was examined. DE induced a significant increase in the nuclear translocation of NF-kappaB (P = 0.02), AP-1 (P = 0.02), phosphorylated JNK (P = 0.04), and phosphorylated p38 (P = 0.01), as well as an increase in total (cytoplasmic + nuclear) immunostaining of phosphorylated p38 (P = 0.03). A significant increase in nuclear phosphorylated tyrosine was also observed (P < 0.05). These observations demonstrate that DE activates redox-sensitive transcription factors in vivo consistent with oxidative stress triggering the increased synthesis of proinflammatory cytokines. PMID- 15749743 TI - Function curve of the membranes that regulate amniotic fluid volume in sheep. AB - Seven singleton 120-day fetal lambs were prepared with a shunt from the lung to the gastric end of the esophagus, a bladder catheter, and multiple amniotic fluid and vascular catheters. The urachus was ligated. Beginning 7 days later, amniotic fluid volumes were determined by drainage, followed by replacement with 1 liter of lactated Ringer (LR) solution. Urine flow into the amnion was measured continuously. In 14 of 27 experiments, amniotic fluid volumes were determined again 2 days after the inflow into the amnion had consisted of urine only and in 13 experiments after the inflow of urine had been supplemented by an intraamniotic infusion of LR solution. Intramembranous absorption was calculated from the inflows and the changes in volume between the beginning and end of each experiment. The relations between absorption rate and amniotic fluid volume, the "function curves," were highly individual. Urine production during the infusion of LR solution did not decrease, fetal plasma renin activity decreased (P < 0.001), and amniotic fluid volume increased by 140% [SE (27%), P < 0.005], but the increase in the amniochorionic absorption rate of 411% [SE (48%), P < 0.001] was greater (P < 0.005) than the increase in volume. Each of the seven fetuses was proven capable of an average intramembranous absorption rate that exceeded 4.5 liters of amniotic fluid per day. During the infusion of LR solution, the increase in the rate of absorption matched the rate of infusion (both in ml/h), with a regression coefficient of 0.75 (P < 0.001). Thus, even for large amniotic fluid volumes, volume is not limited by the absorptive capacity of the amniochorion, and, at least in these preparations, the position of the function curve and not the natural rate of inflow was the major determinant of resting amniotic fluid volume. PMID- 15749744 TI - Leptin receptors are expressed in coronary arteries, and hyperleptinemia causes significant coronary endothelial dysfunction. AB - Obesity is associated with marked increases in plasma leptin concentration, and hyperleptinemia is an independent risk factor for coronary artery disease. As a result, the purpose of this investigation was to test the following hypotheses: 1) leptin receptors are expressed in coronary endothelial cells; and 2) hyperleptinemia induces coronary endothelial dysfunction. RT-PCR analysis revealed that the leptin receptor gene is expressed in canine coronary arteries and human coronary endothelium. Furthermore, immunocytochemistry demonstrated that the long-form leptin receptor protein (ObRb) is present in human coronary endothelium. The functional effects of leptin were determined using pressurized coronary arterioles (<130 microm) isolated from Wistar rats, Zucker rats, and mongrel dogs. Leptin induced pharmacological vasodilation that was abolished by denudation and the nitric oxide synthase inhibitor N(omega)-nitro-l-arginine methyl ester and was absent in obese Zucker rats. Intracoronary leptin dose response experiments were conducted in anesthetized dogs. Normal and obese concentrations of leptin (0.1-3.0 microg/min ic) did not significantly change coronary blood flow or myocardial oxygen consumption; however, obese concentrations of leptin significantly attenuated the dilation to graded intracoronary doses of acetylcholine (0.3-30.0 microg/min). Additional experiments were performed in canine coronary rings, and relaxation to acetylcholine (6.25 nmol/l-6.25 micromol/l) was significantly attenuated by obese concentrations of leptin (625 pmol/l) but not by physiological concentrations of leptin (250 pmol/l). The major findings of this investigation were as follows: 1) the ObRb is present in coronary arteries and coupled to pharmacological, nitric oxide-dependent vasodilation; and 2) hyperleptinemia produces significant coronary endothelial dysfunction. PMID- 15749746 TI - Strain-dependent beta-adrenergic receptor function influences myocardial responses to isoproterenol stimulation in mice. AB - Recently, we showed that compared with the A/J inbred mouse strain, C57BL/6J (B6) mice have an athlete's cardiac phenotype. We postulated that strain differences would result in greater left ventricular (LV) hypertrophy in response to isoproterenol in B6 than A/J mice and tested the hypothesis that a differential response could be explained partly by differences in beta-adrenergic receptor (beta-AR) density and/or coupling. A/J and B6 mice were randomized to receive daily isoproterenol (100 mg/kg sc) or isovolumic vehicle for 5 days. Animals were studied using echocardiography, tail-cuff blood pressure, histopathology, beta-AR density and percent high-affinity binding, and basal and stimulated adenylyl cyclase activities. One hundred twenty-eight mice (66 A/J and 62 B6) were studied. Isoproterenol-treated A/J mice demonstrated greater percent increases in echocardiographic LV mass/body weight (97 +/- 11 vs. 20 +/- 10%, P = 0.001) and in gravimetric heart mass/body weight versus same-strain controls than B6 mice. Histopathology scores (a composite of myocyte hypertrophy, nuclear changes, fibrosis, and calcification) were greater in isoproterenol-treated A/J vs. B6 mice (2.8 +/- 0.2 vs.1.9 +/- 0.3, P < 0.05), as was quantitation of myocyte damage (22.3 +/- 11.5 vs. 4.3 +/- 3.5%). Interstrain differences in basal beta-AR density, high-affinity binding, and adenylyl cyclase activity were not significant. However, whereas isoproterenol-treated A/J mice showed nonsignificant increases in all beta-AR activity measures, isoproterenol-treated B6 mice had lower beta-AR density (57 +/- 6 vs. 83 +/- 8 fmol/mg, P < 0.05), percent high-affinity binding (15 +/- 2 vs. 26 +/- 3%, P < 0.005), and GTP + isoproterenol-stimulated adenylyl cyclase activity (10 +/- 1.1 vs. 5.8 +/- 1.5 pmol cAMP.mg(-1).min(-1)) compared with controls. High-dose, short-term isoproterenol produces greater macro- and microscopic cardiac hypertrophy and injury in A/J than B6 mice. A/J mice, unlike B6 mice, do not experience beta-AR downregulation or uncoupling in response to isoproterenol. Abnormalities in beta adrenergic regulation may contribute to strain-related differences in the vulnerability to isoproterenol-induced cardiac changes. PMID- 15749745 TI - Nitric oxide inhibition unmasks ischemic myocardium-derived vasoconstrictor signals activating endothelin type A receptor of coronary microvessels. AB - NO plays an important role in the compensatory increase in coronary flow conductance against myocardial ischemia, and NO bioavailability is impaired in various diseases. We tested the hypothesis that, when NO production is inhibited, vasoconstrictor signals from the ischemic myocardium are unmasked. We investigated the involvement of endothelin type A (ETA) receptors in the transduction of the constrictor signal. To detect coronary vasoactive signals derived from ischemic myocardium, we used a bioassay system in which an isolated rabbit coronary microvessel (detector vessel, DV) was placed on beating myocardium perfused by the left anterior descending coronary artery (LAD) of an anesthetized open-chest dog (n = 38). The DV was pressurized to 60 cmH2O throughout the experiment and observed with an intravital microscope equipped with a floating objective. After the intrinsic tone of the DV was established, vehicle (n = 7), Nomega-nitro-L-arginine (L-NNA, 100 micromol/l; n = 13), L-NNA + BQ-123 (a selective ETA receptor blocker, 1 micromol/l; n = 7), or BQ-123 alone (1 micromol/l; n = 7) was superfused onto the DV. Thereafter, the LAD of the beating heart was occluded. Coronary occlusion produced significant dilation of the DV by 10 +/- 4%. When L-NNA was applied, the DV significantly constricted by 12 +/- 5% in response to LAD occlusion, and BQ-123 abolished the vasoconstriction. Pretreatment with BQ-123 alone produced an enhancement of the ischemia-induced dilation. We conclude that ischemic myocardium releases transferable vasomotor signals that produce coronary microvascular constriction during the blockade of NO production and the constrictor signal is mediated by ETA receptors. PMID- 15749747 TI - Dynamic autoregulation of cutaneous circulation: differential control in glabrous versus nonglabrous skin. AB - The purpose of this project was to test the hypothesis that, independent of neural control, glabrous and nonglabrous cutaneous vasculature is capable of autoregulating blood flow. In 10 subjects, spectral and transfer function analyses of arterial pressure and skin blood flow (laser-Doppler flowmetry) from glabrous (palm) and nonglabrous (forearm) regions were performed under three conditions: baseline, ganglionic blockade via intravenous trimethaphan administration, and trimethaphan plus oscillatory lower body negative pressure (LBNP; -5 to -10 mmHg) from 0.05 to 0.07 Hz. Oscillatory LBNP was applied to regenerate mean arterial pressure variability that was abolished by ganglionic blockade. Ganglionic blockade was verified by an absence of a heart rate response to a Valsalva maneuver. Spectral power and transfer function gain between blood pressure and skin blood flow were calculated in this oscillatory frequency range (0.05-0.07 Hz). Within this frequency range, ganglionic blockade significantly decreased spectral power of blood flow in both the forearm and palm, whereas regeneration of arterial blood pressure oscillations significantly increased spectral power of forearm blood flow but not palm blood flow. During oscillatory LBNP, transfer function gain between blood pressure and skin blood flow was significantly elevated at the forearm (0.28 +/- 0.03 to 0.53 +/- 0.02 flux units/mmHg; P < 0.05) but was reduced at the palm (4.7 +/- 0.5 to 1.2 +/- 0.1 flux units/mmHg; P < 0.05). These data show that independent of neural control of blood flow, glabrous skin has the ability to buffer blood pressure oscillations and demonstrates a degree of dynamic autoregulation. Conversely, these data suggest that nonglabrous skin has diminished dynamic autoregulatory capabilities. PMID- 15749748 TI - Inhibition of NF-kappaB induces regression of cardiac hypertrophy, independent of blood pressure control, in spontaneously hypertensive rats. AB - The transcription factor nuclear factor (NF)-kappaB plays a leading role in cardiac hypertrophy associated with heart failure, but whether it is involved in cardiac mass reduction is not known. We evaluated whether inhibiting the NF kappaB cascade with pyrrolidine dithiocarbamate (PDTC) in spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto rats (WKYs) affected hypertrophy. We measured NF-kappaB signaling components [NF-kappaB translocation, IkappaBalpha, p65, mRNA and protein levels, and IkappaB kinase-beta (IKKbeta) activity] at 12 and 36 wk in WKYs and SHRs and at 10 wk in PDTC-treated rats (n = 9). NF-kappaB activation was also evaluated in rats treated for 10 wk with captopril or hydralazine alone or with either drug plus PDTC. All components were increased in SHRs compared with WKYs. After PDTC treatment, NF-kappaB activity was inhibited, and heart weight-to-body weight ratio in SHRs was significantly attenuated (3.52 +/- 0.04 to 3.32 +/- 0.05 mg/kg). Captopril treatment significantly reduced cardiac mass (3.5 vs. 3.05 mg/kg; n = 9) and inhibited NF kappaB activity (169.71 +/- 5.70 to 106.7 +/- 12.44). Hydralazine had no effect on cardiac mass (3.5 vs. 3.42 mg/kg) or NF-kappaB activity (169.71 +/- 5.70 to 155.52 +/- 6.11). Hydralazine plus PDTC reduced blood pressure (191.16 +/- 1.7 to 158.5 +/- 2.36 mmHg) and inhibited NF-kappaB activity (169.71 +/- 5.70 to 97.29 +/- 3.65). Our data suggest that 1) cardiac hypertrophy in SHRs is partly due to NF-kappaB activation, 2) inhibition of NF-kappaB activity by PDTC parallels regression of hypertrophy, and 3) regression of hypertrophy is partly due to inhibition of NF-kappaB activity, independent of hypertension. The relationship between NF-kappaB activity and cardiac remodeling is causal, not coincidental. PMID- 15749749 TI - Selective seed abortion affects the performance of the offspring in Bauhinia ungulata. AB - BACKGROUND AND AIMS: Under the microgametophytic competition hypothesis, a non random pattern of seed abortion is expected, in which only the most vigorous seeds reach maturity. In a previous study, it was found that Bauhinia ungulata (Fabaceae) exhibits a pattern of seed abortion dependent on the position of the ovule within the ovary; ovules located in the stylar half of the fruit, close to the point of entry of pollen tubes to the ovary, have a low probability of seed abortion, whereas ovules in the basal half of the fruit are aborted with a high probability. METHODS: A series of experimental fruits was generated, in which ovules from either the stylar (treatments 1 and 2) or the basal (treatments 3 and 4) half of fruits were destroyed, to evaluate whether these patterns of selective seed abortion have an effect on the vigour of the offspring in B. ungulata. KEY RESULTS: Only 53 % of the seed from control fruits germinated. Seed set in fruits from treatments 1 and 2 showed a significantly lower (33-43 %) percentage of germination; the germination of seeds from fruits in treatments 3 and 4 (49-51 %) did not differ from control seeds. In addition, it was found that the differences in vigour of the offspring are not random with respect to the position of the ovule in the pod. CONCLUSIONS: The overall performance of the seeds correlated with their likelihood of maturation. Seeds located at the basal half of the treatment fruits showed lower values of vigour than seeds located on the stylar half. The differences were more marked for early measures of fitness. PMID- 15749750 TI - Phenology of flowering and starch accumulation in grape (Vitis vinifera L.) cuttings and vines. AB - BACKGROUND AND AIMS: A reliable protocol for flowering and fruiting in cuttings was developed with the aim of (a) studying inflorescence and flower development in grapevine cuttings and field plants, and (b) assisting haploid plant production. METHODS: Inflorescence and flower development was studied in 'Gewurztraminer' (GW) and 'Pinot Noir' (PN) grape vines and cuttings grown in a glasshouse, along with variations in starch in the flowers. As there is a strong relationship between flower development and starch, the starch content of reproductive structures was estimated. KEY RESULTS: Inflorescence and flower development were similar in the vines and cuttings with consistent differences between the two cultivars. Indeed, the ontogenesis of male and female organs is not synchronous in GW and PN, with both female and male meiosis occurring earlier in PN than in GW. Moreover, changes of starch reserves were similar in the two plant types. CONCLUSIONS: Cuttings have a similar reproductive physiology to vines, and can be used to study grape physiology and to develop haploid plants. PMID- 15749751 TI - Interaction between Orobanche crenata and its host legumes: unsuccessful haustorial penetration and necrosis of the developing parasite. AB - BACKGROUND AND AIMS: Orobanche species represent major constraints to crop production in many parts of the world as they reduce yield and alter root/shoot allometry. Although much is known about the histology and effect of Orobanche spp. on susceptible hosts, less is known about the basis of host resistance to these parasites. In this work, histological aspects related to the resistance of some legumes to Orobanche crenata have been investigated in order to determine which types of resistance responses are involved in the unsuccessful penetration of O. crenata. METHODS: Samples of resistance reactions against O. crenata on different genotypes of resistant legumes were collected. The samples were fixed, sectioned and stained using different procedures. Sections were observed using a transmission light microscope and by epi-fluorescence. KEY RESULTS: Lignification of endodermal and pericycle host cells seems to prevent parasite intrusion into the root vascular cylinder at early infection stages. But in other cases, established tubercles became necrotic and died. Contrary to some previous studies, it was found that darkening at the infection site in these latter cases does not correspond to death of host tissues, but to the secretion of substances that fill the apoplast in the host-parasite interface and in much of the infected host tissues. The secretions block neighbouring host vessels. This may interfere with the nutrient flux between host and parasite, and may lead to necrosis and death of the developing parasite. CONCLUSIONS: The unsuccessful penetration of O. crenata seedlings into legume roots cannot be attributed to cell death in the host. It seems to be associated with lignification of host endodermis and pericycle cells at the penetration site. The accumulation of secretions at the infection site, may lead to the activation of xylem occlusion, another defence mechanism, which may cause further necrosis of established tubercles. PMID- 15749752 TI - Transpiration, potassium uptake and flow in tobacco as affected by nitrogen forms and nutrient levels. AB - BACKGROUND AND AIMS: Ammonium can result in toxicity symptoms in many plants when it is supplied as the sole source of N. In this work, influences of different nitrogen forms at two levels (2 and 15 mm N) on growth, water relations and uptake and flow of potassium were studied in plants of Nicotiana tabacum 'K 326'. METHODS: Xylem sap from different leaves was collected from 106-d-old tobacco plants cultured in quartz sand by application of pressure to the root system. Whole-shoot transpiration for each of the treatments was measured on a daily basis by weight determination. KEY RESULTS: Total replacement of NO(3)(-)N by NH(4)(+)-N caused a substantial decrease in dry weight gain, even when plants grew under nutrient deficiency. Increasing nutrient concentration resulted in a greater net dry weight gain when nitrogen was supplied as NO(3)(-) or NH(4)NO(3), but resulted in little change when nitrogen was supplied as NH(4)(+). NH(4)(+)-N as the sole N-source also caused reduction in transpiration rate, changes in plant WUE (which depended on the nutrient levels) and a decrease in potassium uptake. However, the amount of xylem-transported potassium in the plants fed with NH(4)(+) was not reduced: it was 457 % or 596 % of the potassium currently taken up at low or high nutrient level, respectively, indicating a massive export from leaves and cycling of potassium in the phloem. CONCLUSIONS: Ammonium reduces leaf stomatal conductance of tobacco plants. The flow and partitioning of potassium in tobacco plants can be changed, depending on the nitrogen forms and nutrient levels. PMID- 15749754 TI - A mechanism of nonphotochemical energy dissipation, independent from PsbS, revealed by a conformational change in the antenna protein CP26. AB - The regulation of light harvesting in higher plant photosynthesis, defined as stress-dependent modulation of the ratio of energy transfer to the reaction centers versus heat dissipation, was studied by means of carotenoid biosynthesis mutants and recombinant light harvesting complexes (LHCs) with modified chromophore binding. The npq2 mutant of Arabidopsis thaliana, blocked in the biosynthesis of violaxanthin and thus accumulating zeaxanthin, was shown to have a lower fluorescence yield of chlorophyll in vivo and, correspondingly, a higher level of energy dissipation, with respect to the wild-type strain and npq1 mutant, the latter of which is incapable of zeaxanthin accumulation. Experiments on purified thylakoid membranes from all three mutants showed that the major source of the difference between the npq2 and wild-type preparations was a change in pigment to protein interactions, which can explain the lower chlorophyll fluorescence yield in the npq2 samples. Analysis of the xanthophyll binding LHC proteins showed that the Lhcb5 photosystem II subunit (also called CP26) undergoes a change in its pI upon binding of zeaxanthin. The same effect was observed in wild-type CP26 upon treatment that leads to the accumulation of zeaxanthin in the membrane and was interpreted as the consequence of a conformational change. This hypothesis was confirmed by the analysis of two recombinant proteins obtained by overexpression of the Lhcb5 apoprotein in Escherichia coli and reconstitution in vitro with either violaxanthin or zeaxanthin. The V and Z containing pigment-protein complexes obtained by this procedure showed different pIs and high and low fluorescence yields, respectively. These results confirm that LHC proteins exist in multiple conformations, an idea suggested by previous spectroscopic measurements (Moya et al., 2001), and imply that the switch between the different LHC protein conformations is activated by the binding of zeaxanthin to the allosteric site L2. The results suggest that the quenching process induced by the accumulation of zeaxanthin contributes to qI, a component of NPQ whose origin was previously poorly understood. PMID- 15749755 TI - Phototropins promote plant growth in response to blue light in low light environments. AB - Phototropins (phot1 and phot2) are plant-specific blue light receptors for phototropism, chloroplast movement, leaf expansion, and stomatal opening. All these responses are thought to optimize photosynthesis by helping to capture light energy efficiently, reduce photodamage, and acquire CO2. However, experimental evidence for the promotion of plant growth through phototropins is lacking. Here, we report dramatic phototropin-dependent effects on plant growth. When plants of Arabidopsis thaliana wild type, the phot1 and phot2 mutants, and the phot1 phot2 double mutant were grown under red light, no significant growth differences were observed. However, if a very low intensity of blue light (0.1 micromol m(-2) s(-1)) was superimposed on red light, large increases in fresh weight up to threefold were found in those plants that carried functional PHOT1 genes. When the intensity of blue light was increased to 1 micromol m(-2) s(-1), the growth enhancement was also found in the phot1 single mutant, but not in the double mutant, indicating that phot2 mediated similar responses as phot1 with a lower sensitivity. The effects occurred under low photosynthetically active radiation in particular. The well-known physiological phototropin-mediated responses, including chloroplast movement, stomatal opening, and leaf expansion, in the different lines tested indicated an involvement of these responses in the blue light-induced growth enhancement. We conclude that phototropins promote plant growth by controlling and integrating a variety of responses that optimize photosynthetic performance under low photosynthetically active radiation in the natural environment. PMID- 15749756 TI - Timing of the maternal-to-zygotic transition during early seed development in maize. AB - In animals, early embryonic development is largely dependent on maternal transcripts synthesized during gametogenesis. Recent data in plants also suggest maternal control over early seed development, but the actual timing of zygotic genome activation is unclear. Here, we analyzed the timing of the maternal-to zygotic transition during early Zea mays seed development. We show that for 16 genes expressed during early seed development, only maternally inherited alleles are detected during 3 d after fertilization in both the embryo and the endosperm. Microarray analyses of precocious embryonic development in apomictic hybrids between maize and its wild relative, Tripsacum, demonstrate that early embryo development occurs without significant quantitative changes to the transcript population in the ovule before fertilization. Precocious embryo development is also correlated with a higher proportion of polyadenylated mRNA in the ovules. Our data suggest that the maternal-to-zygotic transition occurs several days after fertilization. By contrast, novel transcription accompanies early endosperm development, indicating that different mechanisms are involved in the initiation of endosperm and embryo development. PMID- 15749757 TI - Molecular genetic evidence for the role of SGT1 in the intramolecular complementation of Bs2 protein activity in Nicotiana benthamiana. AB - Pepper plants (Capsicum annuum) containing the Bs2 resistance gene are resistant to strains of Xanthomonas campestris pv vesicatoria (Xcv) expressing the bacterial effector protein AvrBs2. AvrBs2 is delivered directly to the plant cell via the type III protein secretion system (TTSS) of Xcv. Upon recognition of AvrBs2 by plants expressing the Bs2 gene, a signal transduction cascade is activated leading to a bacterial disease resistance response. Here, we describe a novel pathosystem that consists of epitope-tagged Bs2-expressing transgenic Nicotiana benthamiana plants and engineered strains of Pseudomonas syringae pv tabaci that deliver the effector domain of the Xcv AvrBs2 protein via the TTSS of P. syringae. This pathosystem has allowed us to exploit N. benthamiana as a model host plant to use Agrobacterium tumefaciens-mediated transient protein expression in conjunction with virus-induced gene silencing to validate genes and to identify protein interactions required for the expression of plant host resistance. In this study, we demonstrate that two genes, NbSGT1 and NbNPK1, are required for the Bs2/AvrBs2-mediated resistance responses but that NbRAR1 is not. Protein localization studies in these plants indicate that full-length Bs2 is primarily localized in the plant cytoplasm. Three protein domains of Bs2 have been identified: the N terminus, a central nucleotide binding site, and a C terminal Leu-rich repeat (LRR). Co-immunoprecipitation studies demonstrate that separate epitope-tagged Bs2 domain constructs interact in trans specifically in the plant cell. Co-immunoprecipitation studies also demonstrate that an NbSGT1 dependent intramolecular interaction is required for Bs2 function. Additionally, Bs2 has been shown to associate with SGT1 via the LRR domain of Bs2. These data suggest a role for SGT1 in the proper folding of Bs2 or the formation of a Bs2 SGT1-containing protein complex that is required for the expression of bacterial disease resistance. PMID- 15749753 TI - Auxin: regulation, action, and interaction. AB - BACKGROUND: The phytohormone auxin is critical for plant growth and orchestrates many developmental processes. SCOPE: This review considers the complex array of mechanisms plants use to control auxin levels, the movement of auxin through the plant, the emerging view of auxin-signalling mechanisms, and several interactions between auxin and other phytohormones. Though many natural and synthetic compounds exhibit auxin-like activity in bioassays, indole-3-acetic acid (IAA) is recognized as the key auxin in most plants. IAA is synthesized both from tryptophan (Trp) using Trp-dependent pathways and from an indolic Trp precursor via Trp-independent pathways; none of these pathways is fully elucidated. Plants can also obtain IAA by beta-oxidation of indole-3-butyric acid (IBA), a second endogenous auxin, or by hydrolysing IAA conjugates, in which IAA is linked to amino acids, sugars or peptides. To permanently inactivate IAA, plants can employ conjugation and direct oxidation. Consistent with its definition as a hormone, IAA can be transported the length of the plant from the shoot to the root; this transport is necessary for normal development, and more localized transport is needed for tropic responses. Auxin signalling is mediated, at least in large part, by an SCFTIR1 E3 ubiquitin ligase complex that accelerates Aux/IAA repressor degradation in response to IAA, thereby altering gene expression. Two classes of auxin-induced genes encode negatively acting products (the Aux/IAA transcriptional repressors and GH3 family of IAA conjugating enzymes), suggesting that timely termination of the auxin signal is crucial. Auxin interaction with other hormone signals adds further challenges to understanding auxin response. CONCLUSIONS: Nearly six decades after the structural elucidation of IAA, many aspects of auxin metabolism, transport and signalling are well established; however, more than a few fundamental questions and innumerable details remain unresolved. PMID- 15749758 TI - The mutant crispa reveals multiple roles for PHANTASTICA in pea compound leaf development. AB - Pinnate compound leaves have laminae called leaflets distributed at intervals along an axis, the rachis, whereas simple leaves have a single lamina. In simple- and compound-leaved species, the PHANTASTICA (PHAN) gene is required for lamina formation. Antirrhinum majus mutants lacking a functional gene develop abaxialized, bladeless adult leaves. Transgenic downregulation of PHAN in the compound tomato (Solanum lycopersicum) leaf results in an abaxialized rachis without leaflets. The extent of PHAN gene expression was found to be correlated with leaf morphology in diverse compound-leaved species; pinnate leaves had a complete adaxial domain of PHAN gene expression, and peltate leaves had a diminished domain. These previous studies predict the form of a compound-leaved phan mutant to be either peltate or an abaxialized rachis. Here, we characterize crispa, a phan mutant in pea (Pisum sativum), and find that the compound leaf remains pinnate, with individual leaflets abaxialized, rather than the whole leaf. The mutant develops ectopic stipules on the petiole-rachis axis, which are associated with ectopic class 1 KNOTTED1-like homeobox (KNOX) gene expression, showing that the interaction between CRISPA and the KNOX gene PISUM SATIVUM KNOTTED2 specifies stipule boundaries. KNOX and CRISPA gene expression patterns indicate that the mechanism of pea leaf initiation is more like Arabidopsis thaliana than tomato. PMID- 15749759 TI - Molecular basis of evolutionary events that shaped the hardness locus in diploid and polyploid wheat species (Triticum and Aegilops). AB - The Hardness (Ha) locus controls grain hardness in hexaploid wheat (Triticum aestivum) and its relatives (Triticum and Aegilops species) and represents a classical example of a trait whose variation arose from gene loss after polyploidization. In this study, we investigated the molecular basis of the evolutionary events observed at this locus by comparing corresponding sequences of diploid, tertraploid, and hexaploid wheat species (Triticum and Aegilops). Genomic rearrangements, such as transposable element insertions, genomic deletions, duplications, and inversions, were shown to constitute the major differences when the same genomes (i.e., the A, B, or D genomes) were compared between species of different ploidy levels. The comparative analysis allowed us to determine the extent and sequences of the rearranged regions as well as rearrangement breakpoints and sequence motifs at their boundaries, which suggest rearrangement by illegitimate recombination. Among these genomic rearrangements, the previously reported Pina and Pinb genes loss from the Ha locus of polyploid wheat species was caused by a large genomic deletion that probably occurred independently in the A and B genomes. Moreover, the Ha locus in the D genome of hexaploid wheat (T. aestivum) is 29 kb smaller than in the D genome of its diploid progenitor Ae. tauschii, principally because of transposable element insertions and two large deletions caused by illegitimate recombination. Our data suggest that illegitimate DNA recombination, leading to various genomic rearrangements, constitutes one of the major evolutionary mechanisms in wheat species. PMID- 15749760 TI - A mitogen-activated protein kinase cascade regulating infection-related morphogenesis in Magnaporthe grisea. AB - Many fungal pathogens invade plants by means of specialized infection structures called appressoria. In the rice (Oryza sativa) blast fungus Magnaporthe grisea, the pathogenicity mitogen-activated protein (MAP) kinase1 (PMK1) kinase is essential for appressorium formation and invasive growth. In this study, we functionally characterized the MST7 and MST11 genes of M. grisea that are homologous with the yeast MAP kinase kinase STE7 and MAP kinase kinase kinase STE11. Similar to the pmk1 mutant, the mst7 and mst11 deletion mutants were nonpathogenic and failed to form appressoria. When a dominant MST7 allele with S212D and T216E mutations was introduced into the mst7 or mst11 mutant, appressorium formation was restored in the resulting transformants. PMK1 phosphorylation also was detected in the vegetative hyphae and appressoria of transformants expressing the MST7(S212D T216E) allele. However, appressoria formed by these transformants failed to penetrate and infect rice leaves, indicating that constitutively active MST7 only partially rescued the defects of the mst7 and mst11 mutants. The intracellular cAMP level was reduced in transformants expressing the MST7(S212D T216E) allele. We also generated MST11 mutant alleles with the sterile alpha motif (SAM) and Ras-association (RA) domains deleted. Phenotype characterizations of the resulting transformants indicate that the SAM domain but not the RA domain is essential for the function of MST11. These data indicate that MST11, MST7, and PMK1 function as a MAP kinase cascade regulating infection-related morphogenesis in M. grisea. Although no direct interaction was detected between PMK1 and MST7 or MST11 in yeast two hybrid assays, a homolog of yeast STE50 in M. grisea directly interacted with both MST7 and MST11 and may function as the adaptor protein for the MST11-MST7 PMK1 cascade. PMID- 15749761 TI - HISTONE DEACETYLASE19 is involved in jasmonic acid and ethylene signaling of pathogen response in Arabidopsis. AB - Histone acetylation is modulated through the action of histone acetyltransferases and deacetylases, which play key roles in the regulation of eukaryotic gene expression. Previously, we have identified a yeast histone deacetylase REDUCED POTASSIUM DEPENDENCY3 (RPD3) homolog, HISTONE DEACETYLASE19 (HDA19) (AtRPD3A), in Arabidopsis thaliana. Here, we report further study of the expression and function of HDA19. Analysis of Arabidopsis plants containing the HDA19:beta glucuronidase fusion gene revealed that HDA19 was expressed throughout the life of the plant and in most plant organs examined. In addition, the expression of HDA19 was induced by wounding, the pathogen Alternaria brassicicola, and the plant hormones jasmonic acid and ethylene. Using green fluorescent protein fusion, we demonstrated that HDA19 accumulated in the nuclei of Arabidopsis cells. Overexpression of HDA19 in 35S:HDA19 plants decreased histone acetylation levels, whereas downregulation of HDA19 in HDA19-RNA interference (RNAi) plants increased histone acetylation levels. In comparison with wild-type plants, 35S:HDA19 transgenic plants had increased expression of ETHYLENE RESPONSE FACTOR1 and were more resistant to the pathogen A. brassicicola. The expression of jasmonic acid and ethylene regulated PATHOGENESIS-RELATED genes, Basic Chitinase and beta-1,3-Glucanase, was upregulated in 35S:HDA19 plants but downregulated in HDA19-RNAi plants. Our studies provide evidence that HDA19 may regulate gene expression involved in jasmonic acid and ethylene signaling of pathogen response in Arabidopsis. PMID- 15749762 TI - Interaction of NIMIN1 with NPR1 modulates PR gene expression in Arabidopsis. AB - The Arabidopsis thaliana NONEXPRESSER OF PR GENES1 (NPR1, also known as NIM1) protein is an essential positive regulator of salicylic acid (SA)-induced PATHOGENESIS-RELATED (PR) gene expression and systemic acquired resistance (SAR). PR gene activity is regulated at the level of redox-dependent nuclear transport of NPR1. NPR1 interacts with members of the TGA family of transcription factors that are known to bind to SA-responsive elements in the PR-1 promoter. In an attempt to identify proteins involved in SA-mediated signal transduction, we previously described the isolation of three novel genes encoding distinct albeit structurally related proteins designated NIMIN1 (for NIM1-INTERACTING1), NIMIN2, and NIMIN3 that interact with NPR1 in the yeast two-hybrid system. Here, we show that NIMIN1 and NPR1 can be copurified from plant extracts, providing biochemical evidence for their interaction. We provide functional evidence for this interaction by describing transgenic plants constitutively expressing high amounts of NIMIN1. These plants show reduced SA-mediated PR gene induction and a compromised SAR, thus mimicking the described phenotype conferred by npr1. Moreover, they showed reduced RESISTANCE gene-mediated protection. These effects were dependent on the ability of NIMIN1 to interact with NPR1. Mutant plants with a T-DNA insertion in NIMIN1 as well as transgenic plants with reduced NIMIN1 mRNA levels showed hyperactivation of PR-1 gene expression after SA treatment but no effect on the disease resistance phenotype. Our results strongly suggest that NIMIN1 negatively regulates distinct functions of NPR1, providing a mechanism to modulate specific features of SAR. PMID- 15749763 TI - The critical role of disulfide bond formation in protein sorting in the endosperm of rice. AB - Many seed storage proteins, including monomeric 2S albumin and polymeric prolamin, contain conserved sequences in three separate regions, termed A, B, and C, which contain the consensus motifs LxxC, CCxQL, and PxxC, respectively. Protein-sorting mechanisms in rice (Oryza sativa) endosperm were studied with a green fluorescent protein (GFP) fused to different segments of rice alpha globulin, a monomeric, ABC-containing storage protein. The whole ABC region together with GFP was efficiently transported to protein storage vacuoles (type II protein bodies [PB-II]) in the endosperm cells and sequestered in the matrix that surrounds the crystalloids. Peptide Gln-23 to Ser-43 in the A region was sufficient to guide GFP to PB-II. However, GFP fused with the AB or B region accumulated in prolamin protein bodies. Substitution mutations in the CCxQL motif in the B region significantly altered protein localization in the endosperm cells. Furthermore, protein extracts containing these substituted proteins had increased amounts of the endoplasmic reticulum (ER) chaperons BiP (for binding protein), protein disulfide isomerase, and calnexin as a part of protein complexes that were insoluble in a detergent buffer. These results suggest that the ER chaperons and disulfide bonds formed at the dicysteine residues in CCxQL play critical roles in sorting fused proteins in the endosperm cells. PMID- 15749765 TI - Molecular basis for the RIN4 negative regulation of RPS2 disease resistance. AB - Recent studies have demonstrated that RPS2, a plasma membrane-localized nucleotide binding site/leucine-rich repeat protein from Arabidopsis thaliana, associates with RPM1 Interacting Protein 4 (RIN4) and that this association functions to modulate the RPS2-mediated defense pathway in response to the bacterial effector protein AvrRpt2. In addition to negatively regulating RPS2 activity, RIN4 is also a target of AvrRpt2, a Cys protease and cognate bacterial effector protein of RPS2. Nicotiana benthamiana has been employed as a heterologous expression system to characterize the RPS2-RIN4 association, defining the domains in RIN4 required for the negative regulation of RPS2 activity. Upon inoculation of N. benthamiana leaves with Agrobacterium tumefaciens expressing RPS2, a rapid hypersensitive response (HR) is detected with 22 h of infiltration. The HR can be blocked by infiltrating the leaf with A. tumefaciens expressing RPS2 in the presence of RIN4, recapitulating the ability of RIN4 to interfere with RPS2-mediated resistance in Arabidopsis. Moreover, in the presence of RIN4, the RPS2-mediated HR can be restored by the delivery of AvrRpt2 via A. tumefaciens. This assay has been developed as a phenotypic marker for (1) the HR-inducing phenotype associated with RPS2, (2) negative regulation of RPS2 by RIN4, and (3) the AvrRpt2-mediated disappearance of RIN4. Here, we present a series of deletion and site-directed mutation analyses to identify amino acids in RIN4 required for the RPS2-RIN4 association and to distinguish these from residues in RIN4 that serve as a target sequence for AvrRpt2. In addition to characterizing the RPS2-RIN4 association in N. benthamiana, we have moved forward to show that the biological relevance of these amino acid changes is applicable in Arabidopsis as well. To this end, we have identified specific amino acids within the C-terminal half of RIN4 that are required for RPS2 regulation and association. PMID- 15749767 TI - Mie-type scattering and non-Beer-Lambert absorption behavior of human cells in infrared microspectroscopy. AB - We report infrared microspectral features of nuclei in a completely inactive and contracted (pyknotic) state, and of nuclei of actively dividing cells. For pyknotic nuclei, the very high local concentration of DNA leads to opaqueness of the chromatin and, consequently, the absence of DNA signals in the IR spectra of very small nuclei. However, these nuclei can be detected by their scattering properties, which can be described by the Mie theory of scattering from dielectric spheres. This scattering depends on the size of the nucleus; consequently, quite different scattering cross-sections are calculated and observed for pyknotic and mitotic nuclei. PMID- 15749768 TI - Comparison of sequence-based and structure-based energy functions for the reversible folding of a peptide. AB - We used computer simulations to compare the reversible folding of a 20-residue peptide, as described by sequence-based and structure-based energy functions. Sequence-based energy functions are transferable and can be used to describe the behavior of different proteins, since interactions are defined between atomic species. Conversely, structure-based energy functions are not transferable, since the interactions are defined relative to the native conformation, which is assumed to correspond to the global minimum of the energy. Our results indicate that the sequence-based and the structure-based descriptions are in qualitative agreement in characterizing the two-state behavior of the peptide that we studied. We also found, however, that several equilibrium properties, including the free-energy landscape, can be significantly different in the various models. These results suggest that the fact that a model describes the native state of a polypeptide chain does not necessarily imply that the thermodynamic and kinetic properties will also be reproduced correctly. PMID- 15749764 TI - Novel functions of plant cyclin-dependent kinase inhibitors, ICK1/KRP1, can act non-cell-autonomously and inhibit entry into mitosis. AB - In animals, cyclin-dependent kinase inhibitors (CKIs) are important regulators of cell cycle progression. Recently, putative CKIs were also identified in plants, and in previous studies, Arabidopsis thaliana plants misexpressing CKIs were found to have reduced endoreplication levels and decreased numbers of cells consistent with a function of CKIs in blocking the G1-S cell cycle transition. Here, we demonstrate that at least one inhibitor from Arabidopsis, ICK1/KRP1, can also block entry into mitosis but allows S-phase progression causing endoreplication. Our data suggest that plant CKIs act in a concentration dependent manner and have an important function in cell proliferation as well as in cell cycle exit and in turning from a mitotic to an endoreplicating cell cycle mode. Endoreplication is usually associated with terminal differentiation; we observed, however, that cell fate specification proceeded independently from ICK1/KRP1-induced endoreplication. Strikingly, we found that endoreplicated cells were able to reenter mitosis, emphasizing the high degree of flexibility of plant cells during development. Moreover, we show that in contrast with animal CDK inhibitors, ICK1/KRP1 can move between cells. On the one hand, this challenges plant cell cycle control with keeping CKIs locally controlled, and on the other hand this provides a possibility of linking cell cycle control in single cells with the supracellular organization of a tissue or an organ. PMID- 15749769 TI - Multiquantum EPR spectroscopy of spin-labeled arrestin K267C at 35 GHz. AB - Three- and five-quantum absorption and dispersion multiquantum electron paramagnetic resonance spectra of a spin-labeled protein have been obtained for the first time at Q-band (35 GHz). Spectra of arrestin spin-labeled at site 267 were recorded at room temperature as a function of microwave power. The separation of irradiating microwave frequencies, Deltaf, was 10 kHz, and a newly designed multiquantum Q-band electron paramagnetic resonance bridge was utilized, operating in a superheterodyne detection mode. The sample volume was 30 nL using a 3-loop-2-gap resonator. Most spectra were obtained at a 300 microM concentration in single, 2-min scans, but spectra were also successfully obtained at 30 microM, corresponding to one picomole of protein. Enhanced sensitivity to T(1) and T(2) was evident in the spectra, and linewidths varied considerably across the spectra. The pure absorption displays are beneficial relative to field modulation methods for spectral characterization. The presence of two states of the nitroxide spin-label with different relaxation times is evident, particularly in the dispersion spectra, which are expected to exhibit enhanced sensitivity to lineshape variation relative to absorption. Feasibility has been established for the use of this technique for site-directed spin-labeling studies of biologically relevant samples, particularly the study of protein structure and dynamics. PMID- 15749766 TI - A novel plant cysteine protease has a dual function as a regulator of 1 aminocyclopropane-1-carboxylic Acid synthase gene expression. AB - The hormone ethylene influences plant growth, development, and some defense responses. The fungal elicitor Ethylene-Inducing Xylanase (EIX) elicits ethylene biosynthesis in tomato (Lycopersicon esculentum) and tobacco (Nicotiana tabacum) leaves by induction of 1-aminocyclopropane-1-caboxylic acid synthase (Acs) gene expression. A minimal promoter element in the LeAcs2 gene required for EIX responsiveness was defined by deletion analysis in transgenic tomato plants. The sequence between -715 and -675 of the tomato Acs2 gene was found to be essential for induction by EIX. A Cys protease (LeCp) was isolated that specifically binds to this cis element in vitro. Ectopic expression of LeCp in tomato leaves induced the expression of Acs2. Moreover, chromatin immunoprecipitation showed that LeCp binds in vivo to the Acs promoter. We propose a mechanism for the dual function of the LeCp protein. The protease acts enzymatically in the cytoplasm. Then, upon signaling, a small ubiquitin-related modifier protein binds to it, enabling entrance into the nucleus, where it acts as a transcription factor. Thus, LeCp can be considered a dual-function protein, having enzymatic activity and, upon elicitor signaling, exhibiting transcriptional factor activity that induces LeAcs2 expression. PMID- 15749770 TI - Single-molecule analysis of epidermal growth factor signaling that leads to ultrasensitive calcium response. AB - Quantitative relationships between inputs and outputs of signaling systems are fundamental information for the understanding of the mechanism of signal transduction. Here we report the correlation between the number of epidermal growth factor (EGF) bindings and the response probability of intracellular calcium elevation. Binding of EGF molecules and changes of intracellular calcium concentration were measured for identical HeLa human epithelial cells. It was found that 300 molecules of EGF were enough to induce calcium response in half of the cells. This number is quite small compared to the number of EGF receptors (EGFR) expressed on the cell surface (50,000). There was a sigmoidal correlation between the response probability and the number of EGF bindings, meaning an ultrasensitive reaction. Analysis of the cluster size distribution of EGF demonstrated that dimerization of EGFR contributes to this switch-like ultrasensitive response. Single-molecule analysis revealed that EGF bound faster to clusters of EGFR than to monomers. This property should be important for effective formation of signaling dimers of EGFR under very small numbers of EGF bindings and suggests that the expression of excess amounts of EGFR on the cell surface is required to prepare predimers of EGFR with a large association rate constant to EGF. PMID- 15749771 TI - Spontaneous formation of detergent micelles around the outer membrane protein OmpX. AB - The structure and flexibility of the outer membrane protein X (OmpX) in a water detergent solution and in pure water are investigated by molecular dynamics simulations on the 100-ns timescale and compared with NMR data. The simulations allow for an unbiased determination of the structure of detergent micelles and the protein-detergent mixed micelle. The short-chain lipid dihexanoylphosphatidylcholine, as a detergent, aggregates into pure micelles of approximately 18 molecules, or alternatively, it binds to the protein surface. The detergent binds in the form of a monolayer ring around the hydrophobic beta barrel of OmpX rather than in a micellar-like oblate; approximately 40 dihexanoylphosphatidylcholine lipids are sufficient for an effective suppression of water from the surface of the beta-barrel region. The phospholipids bind also on the extracellular, protruding beta-sheet. Here, polar interactions between charged amino acids and phosphatidylcholine headgroups act as condensation seed for detergent micelle formation. The polar protein surface remains accessible to water molecules. In total, approximately 90-100 detergent molecules associate within the protein-detergent mixed micelle, in agreement with experimental estimates. The simulation results indicate that OmpX is not a water pore and support the proposed role of the protruding beta-sheet as a "fishing rod". PMID- 15749772 TI - Dielectric control of counterion-induced single-chain folding transition of DNA. AB - In the presence of condensing agents, single chains of giant double-stranded DNA undergo a first-order phase transition between an elongated coil state and a folded compact state. To connect this like-charged attraction phenomenon to counterion condensation, we performed a series of single-chain experiments on aqueous solutions of DNA, where we varied the extent of counterion condensation by varying the relative dielectric constant epsilon(r) from 80 to 170. Single chain observations of changes in the conformation of giant DNA were performed by transmission electron microscopy and fluorescence microscopy, with tetravalent spermine (SPM(4+)) as a condensing agent. At a fixed dielectric constant, single DNA chains fold into a compact state upon the addition of spermine, whereas at a constant spermine concentration single DNA chains unfold with an increase in epsilon(r). In both cases, the transition is largely discrete at the level of single chains. We found that the critical concentration of spermine necessary to induce the single-chain folding transition increases exponentially as the dielectric constant increases, corresponding to 87-88% of the DNA charge neutralized at the onset of the transition. We also observed that the toroidal morphology of compact DNA partially unfolds when epsilon(r) is increased. PMID- 15749773 TI - Nanopore unitary permeability measured by electrochemical and optical single transporter recording. AB - For the analysis of membrane transport processes two single molecule methods are available that differ profoundly in data acquisition principle, achievable information, and application range: the widely employed electrical single channel recording and the more recently established optical single transporter recording. In this study dense arrays of microscopic horizontal bilayer membranes between 0.8 microm and 50 microm in diameter were created in transparent foils containing either microholes or microcavities. Prototypic protein nanopores were formed in bilayer membranes by addition of Staphylococcus aureus alpha-hemolysin (alpha HL). Microhole arrays were used to monitor the formation of bilayer membranes and single alpha-HL pores by confocal microscopy and electrical recording. Microcavity arrays were used to characterize the formation of bilayer membranes and the flux of fluorescent substrates and inorganic ions through single transporters by confocal microscopy. Thus, the unitary permeability of the alpha HL pore was determined for calcein and Ca(2+) ions. The study paves the way for an amalgamation of electrical and optical single transporter recording. Electro optical single transporter recording could provide so far unresolved kinetic data of a large number of cellular transporters, leading to an extension of the nanopore sensor approach to the single molecule analysis of peptide transport by translocases. PMID- 15749774 TI - The SC3 hydrophobin self-assembles into a membrane with distinct mass transfer properties. AB - Hydrophobins are a class of small proteins that fulfill a wide spectrum of functions in fungal growth and development. They do so by self-assembling into an amphipathic membrane at hydrophilic-hydrophobic interfaces. The SC3 hydrophobin of Schizophyllum commune is the best-studied hydrophobin. It assembles at the air water interface into a membrane consisting of functional amyloid fibrils that are called rodlets. Here we examine the dynamics of SC3 assembly at an oil-water and air-water interface and the permeability characteristics of the assembled layer. Hydrophobin assembled at an oil-water interface is a dynamic system capable of emulsifying oil. It accepts soluble-state SC3 oligomers from water in a unidirectional process and sloughs off SC3 vesicles back into the water phase enclosing a portion of the oil phase in their hydrophobic interior. The assembled layer is impermeable to solutes >200 Da from either the water phase or the oil phase; however, due to the emulsification process, oil and the hydrophobic marker molecules in the oil phase can be transferred into the water phase, thus giving the impression that the assembled layer is permeable to the marker molecules. By contrast, the layer assembled at an air-water interface is permeable to water vapor from either the hydrophobic or hydrophilic side. PMID- 15749775 TI - Electrogenic partial reactions of the gastric H,K-ATPase. AB - The fluorescent styryl dye RH421 was used to identify and investigate electrogenic reaction steps of the H,K-ATPase pump cycle. Equilibrium titration experiments were performed with membrane vesicles isolated from hog gastric mucosa, and cytoplasmic and luminal binding of K(+) and H(+) ions was studied. It was found that the binding and release steps of both ion species in both principal conformations of the ion pump, E(1) and P-E(2), are electrogenic, whereas the conformation transitions do not contribute significantly to a charge movement within the membrane dielectric. This behavior is in agreement with the transport mechanism found for the Na,K-ATPase and the sarcoplasmic reticulum Ca ATPase. The data were analyzed on the basis of the Post-Albers reaction cycle. For proton binding, two pK values were found in both conformations: 6.7 and 300-fold) in the spleen. These data show that, in addition to the developmental increase in hypothalamic Ob-Rb expression previously reported, plasma leptin binding activity increases several fold from fetal to adult life in the rat. This suggests that the actions of leptin depend not only on its synthesis in adipose tissue and Ob-Rb expression in target cells, but also on factors that regulate tissue expression of Ob-Re and thus leptin transport within plasma. PMID- 15749813 TI - Spatial distribution of growth hormone receptor, insulin-like growth factor-I receptor and apoptotic chondrocytes during growth plate development. AB - Linear bone growth depends upon proliferation, maturation, and apoptosis of growth plate chondrocytes, processes regulated by growth hormone (GH) and insulin like growth factor-I (IGF-I). To investigate the contribution of GH, IGF-I and apoptosis to growth plate function, the expression of GH receptor (GHR) and IGF-I receptor (IGF-IR) mRNA were evaluated by in situ hybridization in fractionated costochondral growth plates of growing rats (at 2, 4, and 7 weeks). Apoptosis was determined by TUNEL assay and morphology in histological sections. GHR mRNA was greatest in resting cells with hypertropic cells increasing GHR expression with increasing age. Hypertropic and resting cell IGF-IR mRNA declined over the ages studied. Receptor mRNA expression was altered by exposing cells to GH or IGF-I. GH and IGF significantly decreased GHR mRNA in proliferative cells. GH and IGF also decreased IGF-IR mRNA in resting cells and the 2- and 4-week-old proliferative and hypertropic cells. Treating cells in culture with GH increased the number of apoptotic cells across all ages and zones. Histologically, apoptotic cells were observed at the chondro-osseous junction and within actively proliferating chondrocytes but not in resting cells. Apoptosis was highest at 4 weeks of age with lateral regions displaying the greatest number of cells undergoing apoptosis. These data indicate that apoptosis plays a role in growth plate function, particularly spatial configuration as indicated by the preferential lateral cell apoptosis. The susceptibility of proliferative cells to GHR and IGF-IR down regulation during the period of greatest apoptosis supports a role for the GH-IGF axis in both proliferation and apoptosis during growth plate development. PMID- 15749814 TI - Lindane, a gap junction blocker, suppresses FSH and transforming growth factor beta1-induced connexin43 gap junction formation and steroidogenesis in rat granulosa cells. AB - The present study was designed to explore the role of gap junctions in follicle stimulating hormone (FSH) and transforming growth factor beta1 (TGF beta1) stimulated steroidogenesis in ovarian granulosa cells of gonadotropin-primed immature rats. There were three specific aims. First, we investigated the effect of FSH and TGF beta1 as well as lindane (a general gap junction blocker) on the level of connexin43 (Cx43), the major gap junction constituent in granulosa cells, and on gap junction function. The second aim was to determine the effect of lindane on FSH and TGF beta1-stimulated progesterone production and the levels of two critical players, cytochrome P450 side-chain cleavage (P450scc) enzyme and steroidogenic acute regulatory (StAR) protein. The third aim was to further investigate the specific involvement of Cx43 gap junctions in FSH and TGF beta1 stimulated steroidogenesis using a Cx43 mimetic peptide blocker. Immunoblotting analysis showed that FSH plus TGF beta1 dramatically increased the levels of phosphorylated Cx43 without significantly influencing the level of nonphosphorylated Cx43, and this stimulatory effect was completely suppressed by lindane. Also, immunofluorescence analysis showed that Cx43 immuno-reactivity increased in the FSH plus TGF beta1-treated group and predominantly appeared in a punctate pattern at cell-cell contact sites, and lindane reduced such cell periphery immunostaining. Furthermore, TGF beta1 enhanced the FSH-induced gap junction intercellular communication and lindane completely suppressed this effect. In addition, lindane suppressed the FSH and TGF beta1-stimulated increases in progesterone production and the levels of P450scc enzyme and StAR protein. This study demonstrates a clear temporal association between the Cx43 protein level/gap junction communication and progesterone production in rat ovarian granulosa cells in response to FSH and TGF beta1 as well as lindane. Furthermore, a specific Cx43 gap junction blocker suppressed FSH plus TGF beta1 stimulated progesterone production. In conclusion, this study suggests that Cx43 gap junctions may play a critical role in FSH plus TGF beta1-stimulated progesterone production in rat ovarian granulosa cells. PMID- 15749815 TI - Identification, localization and functional in vitro and in vivo activity of oxytocin receptor in the rat penis. AB - We recently found that the oxytocin receptor (OTR) is expressed in the human and rabbit corpus cavernosum and mediates contractility in vitro. The present study extended our investigations to the rat, and explored whether OTR regulates penile detumescence in vivo. Real-time RT-PCR quantitatively characterized the distribution of OTR mRNA in the male genital tract. Specific transcripts for OTR were expressed in all the tissues investigated. Penile expression of OTR was comparable to that observed in testis and prostate. Western blot analysis detected a single band of the expected molecular mass for OTR in all tissues examined, including rat penis. Expression of OTR protein in rat penile extracts was further confirmed by binding studies, using the OTR selective radiolabeled ligand 125I-OTA (K(d) = 17 +/- 6.5 pM, B(max)=15.7 +/- 5 fmoles/mg protein). OTR was immunolocalized to the endothelial and smooth muscle compartments of cavernous spaces and blood vessels. In rat corpus cavernosum strips, oxytocin (OT) and an OTR selective agonist ([Thr4,Gly7]OT) induced identical increases in tension, while different vasopressin agonists were less active. In vivo, OT intra cavernous injection (ICI) dose-dependently inhibited intracavernous pressure (ICP) increase elicited by either electrical stimulation of the cavernous nerve or ICI of papaverine with similar IC(50)s (117.7 +/- 37 mU). The OTR antagonist, atosiban, counteracted the contractile effect of OT both in vitro and in vivo. Atosiban alone significantly increased ICP at lower stimulation frequencies (2 Hz = P<0.001 and 4 Hz = P<0.05 vs control), but not at the maximal frequency (16 Hz). Our data showed that OTR is present in the rat penis and mediates contractility both in vitro and in vivo, therefore suggesting a role for OT in maintaining penile detumescence. PMID- 15749818 TI - Atmospheric brown clouds: impacts on South Asian climate and hydrological cycle. AB - South Asian emissions of fossil fuel SO(2) and black carbon increased approximately 6-fold since 1930, resulting in large atmospheric concentrations of black carbon and other aerosols. This period also witnessed strong negative trends of surface solar radiation, surface evaporation, and summer monsoon rainfall. These changes over India were accompanied by an increase in atmospheric stability and a decrease in sea surface temperature gradients in the Northern Indian Ocean. We conducted an ensemble of coupled ocean-atmosphere simulations from 1930 to 2000 to understand the role of atmospheric brown clouds in the observed trends. The simulations adopt the aerosol radiative forcing from the Indian Ocean experiment observations and also account for global increases in greenhouse gases and sulfate aerosols. The simulated decreases in surface solar radiation, changes in surface and atmospheric temperatures over land and sea, and decreases in monsoon rainfall are similar to the observed trends. We also show that greenhouse gases and sulfates, by themselves, do not account for the magnitude or even the sign in many instances, of the observed trends. Thus, our simulations suggest that absorbing aerosols in atmospheric brown clouds may have played a major role in the observed regional climate and hydrological cycle changes and have masked as much as 50% of the surface warming due to the global increase in greenhouse gases. The simulations also raise the possibility that, if current trends in emissions continue, the subcontinent may experience a doubling of the drought frequency in the coming decades. PMID- 15749817 TI - Syne proteins anchor muscle nuclei at the neuromuscular junction. AB - Vertebrate skeletal muscle fibers contain hundreds of nuclei, of which three to six are functionally specialized and stably anchored beneath the postsynaptic membrane at the neuromuscular junction (NMJ). The mechanisms that localize synaptic nuclei and the roles they play in neuromuscular development are unknown. Syne-1 is concentrated at the nuclear envelope of synaptic nuclei; its Caenorhabditis elegans orthologue ANC-1 functions to tether nuclei to the cytoskeleton. To test the involvement of Syne proteins in nuclear anchoring, we generated transgenic mice overexpressing the conserved C-terminal Klarsicht/ANC 1/Syne homology domain of Syne-1. The transgene acted in a dominant interfering fashion, displacing endogenous Syne-1 from the nuclear envelope. Muscle nuclei failed to aggregate at the NMJ in transgenic mice, demonstrating that localization and positioning of synaptic nuclei require Syne proteins. We then exploited this phenotype to show that synaptic nuclear aggregates are dispensable for maturation of the NMJ. PMID- 15749819 TI - The cellular HIV-1 Rev cofactor hRIP is required for viral replication. AB - An important goal of contemporary HIV type 1 (HIV-1) research is to identify cellular cofactors required for viral replication. The HIV-1 Rev protein facilitates the cytoplasmic accumulation of the intron-containing viral gag-pol and env mRNAs and is required for viral replication. We have previously shown that a cellular protein, human Rev-interacting protein (hRIP), is an essential Rev cofactor that promotes the release of incompletely spliced HIV-1 RNAs from the perinuclear region. Here, we use complementary genetic approaches to ablate hRIP activity and analyze HIV-1 replication and viral RNA localization. We find that ablation of hRIP activity by a dominant-negative mutant or RNA interference inhibits virus production by mislocalizing Rev-directed RNAs to the nuclear periphery. We further show that depletion of endogenous hRIP by RNA interference results in the loss of viral replication in human cell lines and primary macrophages; virus production was restored to wild-type levels after reintroduction of hRIP protein. Taken together, our results indicate that hRIP is an essential cellular cofactor for Rev function and HIV-1 replication. Because hRIP is not required for cell viability, it may be an attractive target for the development of new antiviral strategies. PMID- 15749820 TI - Transcriptional network underlying Caenorhabditis elegans vulval development. AB - The vulval development of Caenorhabditis elegans provides an opportunity to investigate genetic networks that control gene expression during organogenesis. During the fourth larval stage (L4), seven vulval cell types are produced, each of which executes a distinct gene expression program. We analyze how the expression of cell-type-specific genes is regulated. Ras and Wnt signaling pathways play major roles in generating the spatial pattern of cell types and regulate gene expression through a network of transcription factors. One transcription factor (lin-29) primarily controls the temporal expression pattern. Other transcription factors (lin-11, cog-1, and egl-38) act in combination to control cell-type-specific gene expression. The complexity of the network arises in part because of the dynamic nature of gene expression, in part because of the presence of seven cell types, and also because there are multiple regulatory paths for gene expression within each cell type. PMID- 15749822 TI - Mechanical unfolding of RNA hairpins. AB - Mechanical unfolding trajectories, generated by applying constant force in optical-tweezer experiments, show that RNA hairpins and the P5abc subdomain of the group I intron unfold reversibly. We use coarse-grained Go-like models for RNA hairpins to explore forced unfolding over a broad range of temperatures. A number of predictions that are amenable to experimental tests are made. At the critical force, the hairpin jumps between folded and unfolded conformations without populating any discernible intermediates. The phase diagram in the force temperature (f, T) plane shows that the hairpin unfolds by an all-or-none process. The cooperativity of the unfolding transition increases dramatically at low temperatures. Free energy of stability, obtained from time averages of mechanical unfolding trajectories, coincides with ensemble averages, which establishes ergodicity. The hopping time between the native basin of attraction (NBA) and the unfolded basin increases dramatically along the phase boundary. Thermal unfolding is stochastic, whereas mechanical unfolding occurs in "quantized steps" with great variations in the step lengths. Refolding times, upon force quench, from stretched states to the NBA are at least an order of magnitude greater than folding times by temperature quench. Upon force quench from stretched states, the NBA is reached in at least three stages. In the initial stages, the mean end-to-end distance decreases nearly continuously, and there is a sudden transition to the NBA only in the last stage. Because of the generality of the results, we propose that similar behavior should be observed in force quench refolding of proteins. PMID- 15749821 TI - Lipofuscin accumulation, abnormal electrophysiology, and photoreceptor degeneration in mutant ELOVL4 transgenic mice: a model for macular degeneration. AB - Macular degeneration is a heterogeneous group of disorders characterized by photoreceptor degeneration and atrophy of the retinal pigment epithelium (RPE) in the central retina. An autosomal dominant form of Stargardt macular degeneration (STGD) is caused by mutations in ELOVL4, which is predicted to encode an enzyme involved in the elongation of long-chain fatty acids. We generated transgenic mice expressing a mutant form of human ELOVL4 that causes STGD. In these mice, we show that accumulation by the RPE of undigested phagosomes and lipofuscin, including the fluorophore, 2-[2,6-dimethyl-8-(2,6,6-trimethyl-1-cyclohexen-1-yl) 1E,3E,5E,7E-octatetraenyl]-1-(2-hyydroxyethyl)-4-[4-methyl-6-(2,6,6,-trimethyl-1 cyclohexen-1-yl)-1E,3E,5E-hexatrienyl]-pyridinium (A2E) is followed by RPE atrophy. Subsequently, photoreceptor degeneration occurs in the central retina in a pattern closely resembling that of human STGD and age-related macular degeneration. The ELOVL4 transgenic mice thus provide a good model for both STGD and dry age-related macular degeneration, and represent a valuable tool for studies on therapeutic intervention in these forms of blindness. PMID- 15749823 TI - Multi-component protein complexes and Golgi membrane trafficking. AB - Several large cytosolic protein complexes with multiple components have been proposed to play key roles in mediating or controlling membrane trafficking. Among these complexes, TRAPP, COG and GARP/VFT have been implicated in multiple steps of Golgi membrane trafficking. The importance of these complexes for Golgi function has been established using in vitro biochemical assays and yeast and mammalian somatic cell genetics. Furthermore, mutations in the genes encoding subunits of either TRAPP or the COG complex have been shown to be responsible for human genetic disorders. We here review recent studies exploring the structures and functions of these three oligomeric complexes. PMID- 15749824 TI - p97/p47-Mediated biogenesis of Golgi and ER. AB - In mammalian cells, the Golgi apparatus and endoplasmic reticulum have typical structures during interphase: stacked cisternae located adjacent to the nucleus and a network of interconnected tubules throughout the cytoplasm, respectively. At mitosis their architectures disappear and are reassembled in daughter cells. p97, an AAA-ATPase, mediates membrane fusion and is required for reassembly of these organelles. In the p97-mediated membrane fusion, p47 was identified as an essential cofactor, through which p97 binds to a SNARE, syntaxin5. A second essential cofactor, VCIP135, was identified as a p97/p47/syntaxin5-interacting protein. Several lines of recent evidence suggest that ubiquitination may be implicated in the p97/p47 pathway; p47 binds to monoubiquitinated proteins and VCIP135 shows a deubiquitinating activity in vitro. For the cell-cycle regulation of the p97/p47 pathway, it has been reported that the localization and phosphorylation-dephosphorylation of p47 are crucial. In this review, we describe the components involved in the p97-mediated membrane fusion and discuss the regulation of the fusion pathway. PMID- 15749825 TI - Application of quantitative real-time PCR for monitoring the process of enrichment of clones on in vitro protein selection. AB - In vitro selection of proteins from cDNA libraries using display technologies, such as the in vitro virus method, is a powerful means for the discovery of novel protein interactions. After iterative screening, selected proteins are usually identified and evaluated by cloning and sequencing analysis. Previously we applied real-time PCR for evaluation of the sequences obtained on in vitro virus screening. Here, we have presented additional data regarding monitoring of the process of enrichment of selected clones in each round of selection and elimination of false positives by real-time PCR, and have also discussed the utility of the novel method. This approach should also be applicable to other display technologies. PMID- 15749826 TI - X-linked inhibitor of apoptosis functions as ubiquitin ligase toward mature caspase-9 and cytosolic Smac/DIABLO. AB - Members of the IAP (inhibitor of apoptosis) family function as anti-apoptotic proteins by binding directly to caspase-3, -7, and -9 to inhibit their activities. During apoptosis, the activities of IAPs are relieved by a second mitochondria-derived caspase activator, named Smac/DIABLO. Some IAPs have a C terminal RING finger domain that has been identified as the essential motif for the activity of ubiquitin ligase (E3). Here we show that X-linked IAP (XIAP) mediates the polyubiquitination of caspase-9 and Smac. The large subunit of mature caspase-9 was polyubiquitinated by XIAP in vitro, while procaspase-9 was not. Furthermore, the polyubiquitinated form of caspase-9 accumulated in an XIAP dependent manner in intact cells. The ubiquitination of caspase-9 was significantly inhibited in the presence of mature Smac, whereas XIAP was also found to promote the polyubiquitination of cytosolic Smac both in vitro and in intact cells. These ubiquitination reactions require the RING finger domain of XIAP. These findings suggest that XIAP functions as ubiquitin ligase toward mature caspase-9 and Smac to inhibit apoptosis. PMID- 15749827 TI - A novel UbcH10-binding protein facilitates the ubiquitinylation of cyclin B in vitro. AB - UbcH10 is known to act as a ubiquitin-conjugating enzyme (E2) for anaphase promoting complex/cyclosome. Since some E2s support different ubiquitin ligases (E3), it is possible that UbcH10 interacts with other proteins. We cloned a novel protein named H10BH by using a yeast two-hybrid screening method with UbcH10 as bait. The carboxyl terminus of H10BH showed a weak homology to the HECT (homologous to E6-AP carboxyl terminus) domain, which is conserved in one of the families of E3. H10BH bound UbcH10, and the amino acid sequence between 235 and 257 was necessary for this binding. H10BH showed a self-ubiquitinylation activity in a HECT-like sequence-dependent manner. The carboxyl terminal half (amino acids 188-389) showed stronger activity than the full-length H10BH. Furthermore, the carboxyl terminal half of H10BH was able to bind cyclin B and ubiquitinylate cyclin B in vitro. These results suggest that H10BH functions as an E3 using UbcH10 for its E2. PMID- 15749828 TI - Mutagenesis of the active site lysine 221 of the pyruvate kinase from Bacillus stearothermophilus. AB - Lysine 221 of the pyruvate kinase from Bacillus stearothermophilus was mutated to arginine, leucine, asparatic acid and cysteine. All the mutated enzymes were 10(4) to 10(5) times less active than the wild-type enzyme. The cysteine-free enzyme C9S/C268S, and the enzyme C9S/C268S/K221C, which possessed a unique sulfhydryl group at position 221, were prepared. The former had comparable activity to the wild-type enzyme and the latter was 10(4) times less active. These enzymes were denatured and renatured after aminoethylation. The C9S/C268S/K221C enzyme failed to regain its activity when renatured without aminoethylation; but when it was renatured after aminoethylation, it regained 4.5% of the activity of the C9S/C268S enzyme. This evidence suggests the importance of the Lys221 for the pyruvate kinase activity. The kinetic parameters of the S-aminoethylated C9S/C268S/K221C enzyme suggest that it has decreased affinity for phosphoenolpyruvate. PMID- 15749829 TI - Interaction of N-terminal acetyltransferase with the cytoplasmic domain of beta amyloid precursor protein and its effect on A beta secretion. AB - The processing of beta-amyloid precursor protein (APP) generates the amyloid beta protein (A beta) and contributes to the development of Alzheimer's disease (AD). Elucidating the regulation of APP processing will, therefore, contribute to the understanding of AD. Many APP-binding proteins, such as FE65, X11s, and JNK interacting proteins (JIPs), bind the motif 681-GYENPTY-687 within the cytoplasmic domain of APP. Here we found that the human homologue of yeast amino terminal acetyltransferase ARD1 (hARD1) interacts with a novel motif, 658-HGVVEVD 664, in the cytoplasmic domain of APP695. hARD1 expressed its acetyltransferase activity in association with a human subunit homologous to another yeast amino acetyltransferase, hNAT1. Co-expression of hARD1 and hNAT1 in cells suppressed A beta40 secretion and the suppression correlated with their enzyme activity. These observations suggest that the association of APP with hARD1 and hNAT1 and/or their N-acetyltransferase activity contributes to the regulation of A beta generation. PMID- 15749830 TI - Cleavage of nonmuscle myosin heavy chain-A during apoptosis in human Jurkat T cells. AB - We have previously reported that calpastatin, an endogenous inhibitory protein of calpain, is cleaved by a caspase-3-like protease during apoptosis in human Jurkat T cells [Kato, M. et al. (2000) J. Biochem. 127, 297-305]. In this study, we found that nonmuscle myosin heavy chain-A (NMHC-A) is cleaved during apoptosis in Jurkat cells by using a cleavage-site-directed antibody for calpastatin. The cleavage-site-directed antibody was raised against the amino-terminal fragment of calpastatin, and this antibody detected the in vitro cleaved calpastatin fragment. Although cleaved calpastatin was not detected, a 95-kDa polypeptide (p95) was detected in apoptotic cells by this antibody. This p95 was identified as the carboxyl-terminal fragment of NMHC-A based on the results of peptide mass spectrometry fingerprinting and amino-terminal sequencing. Furthermore, two cleavage sites on NMHC-A, Asp-1153 and Asp-1948, were determined, and three cleaved fragments of NMHC-A, one cleaved at Asp-1153 and the other two cleaved at Asp-1948, were detected by cleavage-site-directed antibodies against each cleavage site. The results of confocal immunofluorescence microscopic analysis show that the cleavage at Asp-1948 occurs faster than that at Asp-1153 during apoptosis. In addition, the Asp-1153 cleaved fragment was distributed diffusely in the cytoplasm of apoptotic cells, whereas the Asp-1948 cleaved fragments were detected as condensed dots. In conclusion, our findings can be summarized as follows: (i) NMHC-A is cleaved at two sites during apoptosis, (ii) the timing of cleavage is different between these two cleavage sites, and (iii) the distribution of cleaved fragments is different in apoptotic cells. PMID- 15749831 TI - Identification and characterization of a mouse dipeptidase that hydrolyzes L carnosine. AB - L-Carnosine is a bioactive dipeptide present in mammalian tissues including the central nervous system. We have recently shown that L-carnosine is involved in the regulation of energy homeostasis through the autonomic nervous system, but the mechanisms for its biosynthesis and degradation have not yet been fully elucidated. Here we report the biochemical and immunohistochemical characterization of a mammalian protein that has a 17% overall amino acid sequence homology with a Lactobacilus carnosinase, PepV. A recombinant protein expressed in E. coli has the enzymatic ability to digest L-carnosine and various other dipeptides, and this activity is inhibited by bestatin. It requires Mn2+ for enzymatic activity and its effect is reversible. Immunohistochemical analysis showed that a few neuronal populations express this protein at very high levels. It is highly expressed in the parafascicular nucleus of the thalamus, tuberomammillary nucleus of the hypothalamus and the mitral cell layer of the olfactory bulb. In addition, neuronal processes, but not cell bodies, are stained in the striatum. In all these areas, the protein did not colocalize with the glial fibrilary acidic protein. These results suggest that a peptidase that digests L-carnosine is enriched in several specific neuronal populations in the central nervous system. PMID- 15749832 TI - Molecular mechanisms involved in robustness of yeast central metabolism against null mutations. AB - Adaptive strategies employed by the yeast Saccharomyces cerevisiae provide robustness and adaptability of its central metabolism. Since central metabolism in yeast has been well studied at the enzymatic and genetic levels, it represents an excellent system for evaluating the relative roles of duplicate genes and alternative metabolic pathways as possible mechanisms for the stability of central metabolism against null mutations. Yeast appears to employ a variety of mechanisms to ensure functional robustness of its central metabolism. Uninterrupted flow of energy and precursor metabolites through the pathways of central metabolism via glycolysis (EMP), pentose phosphate shunt (PPS), and the tricarboxylic acid (TCA) cycle are ensured by a variety of adaptive mechanisms. One of the most significant mechanisms appears to be gene duplication events that have produced a number of isozymes functioning under variable environmental and physiological conditions. Alternative pathways represent another important mechanism for increasing the robustness of the system. The robustness of the pathways of central metabolism is apparently higher than that of the other parts of metabolism, because of its exceptional importance to the organism's vitality. The proportion of duplicated viable genes also is substantially larger in central metabolism than that in a pool of other metabolic genes. PMID- 15749833 TI - Functional role of c-Src in IL-1-induced NF-kappa B activation: c-Src is a component of the IKK complex. AB - Interleukin-1 (IL-1) mediates numerous host responses through the rapid activation of nuclear factor-kappa B (NF-kappa B), but the signal pathways leading to NF-kappa B activation are regulated at multiple stages. Here, we propose a novel regulatory system for IL-1-induced NF-kappa B activation by a tyrosine kinase, c-Src. The kinase activity of c-Src increases in an IL-1 dependent manner and the ectopic expression of c-Src augments IL-1-induced NF kappa B activation, suggesting the involvement of c-Src in IL-1 signaling. However, a Src family inhibitor, PP2 failed to inhibit IL-1-induced NF-kappa B activation, and the expression of a c-Src mutant lacking kinase activity (c-Src KD) augmented IL-1-induced NF-kappa B activation as well as wild type c-Src, indicating that the tyrosine kinase activity is not required for IL-1-induced NF kappa B activation. Furthermore, a physiological interaction between c-Src and I kappa B kinase gamma (IKK gamma) was observed, implying the involvement of c-Src in the IKK-complex. While c-Src augmented IL-1-induced IKK activation independent of its kinase activity, the region comprising amino acids 361-440 in the c-Src kinase domain are required for NF-kappa B activation. The same region of c-Src is also required for IL-1-induced IKK activation and the association with IKK gamma. Taken together, our results suggest that c-Src plays a critical role in IL-1 induced NF-kappa B activation through the IKK complex. PMID- 15749834 TI - D-Glutamic acid-induced muscle contraction in the silkworm, Bombyx mori. AB - Agonists for muscle contraction in silkworms were screened by injecting test solutions into the hemolymph of decapitated silkworm larvae. Kainic acid, a glutamate receptor agonist, and D-glutamic acid induced muscle contractions, and D-aspartic acid was partially effective, whereas NMDA and AMPA, representative mammalian glutamate receptor agonists, did not induce contraction. L-Glutamic acid inhibited the kainic acid or D-glutamic acid-induced contraction. Amino acid analysis revealed that 3% of the total glutamic acid in the silkworm hemolymph is D-glutamic acid. These results suggest that d-glutamic acid acts physiologically as an agonist for muscle contraction in silkworms, and that L-glutamic acid functions as an inhibitor. PMID- 15749835 TI - Identification and characterization of the Hesr1/Hey1 as a candidate trans-acting factor on gene expression through the 3' non-coding polymorphic region of the human dopamine transporter (DAT1) gene. AB - Expression of the dopamine transporter (DAT) gene affects dopaminergic neurotransmission. A variable number tandem repeat (VNTR) polymorphism in the 3' non-coding region of the human DAT (DAT1) gene has been reported to affect gene expression as a cis-element, and is associated with some neuropsychiatric disorders. In this study, we identified the basic helix-loop-helix (bHLH) transcriptional factor Hesr1 (the Hairy/enhancer of split related transcriptional factor 1 with a YRPW motif, also named Hey1/HERP2/HRT1/CHF2) as a trans-acting factor in a yeast one-hybrid system, and showed that Hesr1 down-regulates reporter gene expression with the 3' non-coding region of DAT1 gene in mammalian cell lines. The negative regulations depend on bHLH and the Orange domain of the molecule, but not the YRPW motif. The negative regulations affect the VNTR dependent differences in gene expression. In addition, we identified a splice variant, Hesr1-12nt, with a lower activity. We also show that SNP of Hesr1, C386A, causes a lack of activity. These results suggest that Hesr1 and its polymorphism(s) might be involved in dopamine-related polygenic disorders and behavioral traits. PMID- 15749836 TI - Endogenous cannabinoid receptor ligand induces the migration of human natural killer cells. AB - 2-Arachidonoylglycerol is an endogenous ligand for the cannabinoid receptors (CB1 and CB2). Evidence is gradually accumulating which shows that 2 arachidonoylglycerol plays important physiological roles in several mammalian tissues and cells, yet the details remain ambiguous. In this study, we first examined the effects of 2-arachidonoylglycerol on the motility of human natural killer cells. We found that 2-arachidonoylglycerol induces the migration of KHYG 1 cells (a natural killer leukemia cell line) and human peripheral blood natural killer cells. The migration of natural killer cells induced by 2 arachidonoylglycerol was abolished by treating the cells with SR144528, a CB2 receptor antagonist, suggesting that the CB2 receptor is involved in the 2 arachidonoylglycerol-induced migration. In contrast to 2-arachidonoylglycerol, anandamide, another endogenous cannabinoid receptor ligand, did not induce the migration. Delta9-tetrahydrocannabinol, a major psychoactive constituent of marijuana, also failed to induce the migration; instead, the addition of delta9 tetrahydrocannabinol together with 2-arachidonoylglycerol abolished the migration induced by 2-arachidonoylglycerol. It is conceivable that the endogenous ligand for the cannabinoid receptor, that is, 2-arachidonoylglycerol, affects natural killer cell functions such as migration, thereby contributing to the host-defense mechanism against infectious viruses and tumor cells. PMID- 15749837 TI - Factor H is a dermatan sulfate-binding protein: identification of a dermatan sulfate-mediated protease that cleaves factor H. AB - Dermatan sulfate mediates the blood coagulation cascade by binding to heparin cofactor II and potentiating the antithrombin activity. In order to explore another function of dermatan sulfate, a dermatan sulfate affinity column was prepared from biotinylated dermatan sulfate and Streptavidin Sepharose. When human plasma was applied on the dermatan sulfate column, factor H was bound and cleaved. The cleavage products, a 30-kDa N-terminal fragment and a 120-kDa fragment, were eluted from the column with 500 mM NaCl and detected after Western blotting with anti-factor H. The bond between the tandem arginine residues in the sixth domain of factor H was cleaved. When purified factor H was applied on the column, the factor H was not cleaved and was recovered from the column as an intact 150-kDa fraction. The finding that dermatan sulfate-mediated cleavage of factor H was inhibited by (p-amidinophenyl) methanesulfonyl fluoride, but not N ethylmaleimide or EDTA, indicates that a serine protease in the plasma was activated on the dermatan sulfate column and factor H was cleaved without intervention of the plasma protease inhibitors. Amidase activity was detected in the effluent from the dermatan sulfate column but was abolished by pretreatment of the plasma with dermatan sulfate. Therefore, dermatan sulfate participates in the activation of a protease as well as having the protease inhibitory action. PMID- 15749838 TI - Family 19 chitinase from Aeromonas sp. No.10S-24: role of chitin-binding domain in the enzymatic activity. AB - Family 19 chitinase from Aeromonas sp. No.10S-24 (72.6 kDa) is composed of two chitin-binding domains (ChBDs), two proline- and threonine-rich (PT-rich) linkers, and a catalytic domain. The purified enzyme was labile in a standard buffer condition and spontaneously degraded into a 46-kDa fragment upon storage at 4 degrees C. The N-terminal sequence of the 46-kDa fragment was found to correspond to the sequence of the C-terminal region of the second PT-rich linker, indicating that the 46-kDa fragment is produced by truncation of the two ChBDs and the two PT-rich linkers from the mature protein, and consists only of the catalytic domain. The hydrolytic activities toward insoluble and soluble substrates were significantly reduced by the truncation of two ChBDs. In addition, antifungal activity determined from the digestion rate of haustoria of powdery mildew was reduced by the ChBD truncation. Although the profile of the time-course of N-acetylglucosamine hexasaccharide [(GlcNAc)6] degradation catalyzed by the ChBD-truncated enzyme was similar to that of the mature enzyme protein, the specific activity of the ChBD-truncated enzyme determined from the rate of hexasaccharide degradation was lower than that of the mature enzyme. The two CBDs appear to be responsible for facilitating the hydrolytic reaction. The sugar residue affinities (binding free energy changes) at the individual subsites, (-2) (-1) (+1) (+2) (+3) (+4), were estimated by modeling the hexasaccharide hydrolysis by the mature and ChBD-truncated enzymes. The truncation of ChBDs was found to strongly affect the affinity at the (-1) site. This situation seems to result in the lower enzymatic activity of the ChBD truncated enzyme toward the chitinous substrates. PMID- 15749839 TI - Leptin in immunology. AB - Leptin is an adipokine which conveys information on energy availability. In humans, leptin influences energy homeostasis and regulates neuroendocrine function primarily in states of energy deficiency. As a cytokine, leptin also affects thymic homeostasis and, similar to other proinflammatory cytokines, leptin promotes Th1 cell differentiation and cytokine production. We review herein recent advances on the role of leptin in the pathophysiology of immune responses. PMID- 15749840 TI - The prevalence of regulatory T cells in lymphoid tissue is correlated with viral load in HIV-infected patients. AB - Inadequate local cell-mediated immunity appears crucial for the establishment of chronic HIV infection. Accumulation of regulatory T cells (Treg) at the site of HIV replication, the lymphoid organs, may influence the outcome of HIV infection. Our data provide the first evidence that chronic HIV infection changes Treg tissue distribution. Several molecules characteristics of Treg (FoxP3, CTLA-4, glucocorticoid-induced TNFR family-related receptor, and CD25) were expressed more in tonsils of untreated patients compared with antiretroviral-treated patients. Importantly, most FoxP3+ cells expressed CTLA-4, but not CD69. Furthermore, a direct correlation between FoxP3 levels and viral load was evident. In contrast, FoxP3 expression was decreased in circulating T cells from untreated patients, but normalized after initiation of treatment. Functional markers of Treg activity (indoleamine 2,3-dioxygenase, TGF-beta, and CD80) were markedly increased in the tonsils of untreated patients. Our data could provide a new basis for immune-based therapies that counteract in vivo Treg and thereby reinforce appropriate antiviral immunity. PMID- 15749841 TI - IL-10-independent STAT3 activation by Toxoplasma gondii mediates suppression of IL-12 and TNF-alpha in host macrophages. AB - Infection of mouse macrophages by Toxoplasma gondii renders the cells resistant to proinflammatory effects of LPS triggering. In this study, we show that cell invasion is accompanied by rapid and sustained activation of host STAT3. Activation of STAT3 did not occur with soluble T. gondii extracts or heat-killed tachyzoites, demonstrating a requirement for live parasites. Parasite-induced STAT3 phosphorylation and suppression of LPS-triggered TNF-alpha and IL-12 was intact in IL-10-deficient macrophages, ruling out a role for this anti inflammatory cytokine in the suppressive effects of T. gondii. Most importantly, Toxoplasma could not effectively suppress LPS-triggered TNF-alpha and IL-12 synthesis in STAT3-deficient macrophages. These results demonstrate that T. gondii exploits host STAT3 to prevent LPS-triggered IL-12 and TNF-alpha production, revealing for the first time a molecular mechanism underlying the parasite's suppressive effect on macrophage proinflammatory cytokine production. PMID- 15749842 TI - Signaling lymphocytic activation molecule-associated protein controls NKT cell functions. AB - X-linked lymphoproliferative disease (XLP) is a fatal immunological disorder that typically manifests following EBV infection. XLP patients exhibit a number of immune defects including abnormal T, B, and NK lymphocyte function. These defects have been attributed to mutations of Src homology 2 domain-containing gene 1A (SH2D1A), the gene encoding signaling lymphocytic activation molecule-associated protein (SAP), an intracellular adaptor molecule expressed in lymphocytes. We have observed that SAP knockout (SAPKO) mice and humans with XLP have a complete lack of CD1d-restricted NKT cells. As expected, SAPKO mice injected with the NKT cell agonist, alpha-galactosylceramide failed to generate NKT cell IFN-gamma or IL-4. Furthermore, in contrast to wild-type littermates, SAPKO mice coinjected with OVA and alpha-galactosylceramide failed to mount OVA-specific CTL responses. These data suggest that an absence of NKT cells may underlie part of the immune dysregulation seen in SAPKO mice and in XLP patients. PMID- 15749843 TI - Recent immune status determines the source of antigens that drive homeostatic T cell expansion. AB - Homeostatic proliferation of naive T cells transferred to T cell-deficient syngeneic mice is driven by low-affinity self-MHC/peptide ligands and the cytokine IL-7. In addition to homeostatic proliferation, a subset of naive T cells undergoes massive proliferation in chronically immunodeficient hosts, but not in irradiated normal hosts. Such rapid T cell proliferation occurs largely independent of homeostatic factors, because it was apparent in the absence of IL 7 and in T cell-sufficient hosts devoid of functional T cell immunity. Strikingly, immunodeficient mice raised under germfree conditions supported only slow homeostatic proliferation, but not the marked T cell proliferation observed in conventionally raised immunodeficient mice. Thus, polyclonal naive T cell expansion in T cell-deficient hosts can be driven predominantly by either self Ags or foreign Ags depending on the host's previous state of T cell immunocompetency. PMID- 15749844 TI - Stat5 mediates the IL-7-induced accessibility of a representative D-Distal VH gene. AB - During B cell development, discrete domains within the Igh locus are activated independently for recombination. The D-distal V(H) genes are uniquely dependent on IL-7R signaling, which is thought to establish local chromatin accessibility through an unknown mechanism. To dissect this mechanism, we used a murine B cell line that responds to IL-7 by specifically inducing accessibility of a representative D-distal gene (A1) but not a D-proximal gene (V11). We demonstrate that IL-7-activated Stat5 is recruited rapidly to the A1 gene, with a concomitant increase in germline transcription and H4 acetylation. Furthermore, retroviral transduction of dominant negative or constitutively active Stat5 demonstrated that Stat5 activation is both necessary and sufficient for the IL-7-induced A1 germline transcription. Lastly, as with all known Stat5 target genes, A1 germline transcription requires a deacetylase activity. These results demonstrate that in response to IL-7 signaling, Stat5 is recruited to the D-distal A1 gene and induces accessibility. PMID- 15749845 TI - Novel role of lipoxygenases in the inflammatory response: promotion of TNF mRNA decay by 15-hydroperoxyeicosatetraenoic acid in a monocytic cell line. AB - The metabolism of arachidonic acid via the lipoxygenase and cyclooxygenase pathways generates metabolites that regulate the inflammatory response. Although products of lipoxygenase are classically proinflammatory, recently it has been demonstrated that lipoxins, 15-hydroperoxyeicosatetraenoic acid (15-HPETE) and 15 hydroxyeicosatetraenoic acid exhibit anti-inflammatory activity. We now demonstrate for the first time that 15-HPETE regulates the production of the proinflammatory cytokine TNF posttranscriptionally by promoting degradation of LPS-induced TNFmRNA in a human monocytic cell line, Mono Mac 6. 15-HPETE causes a significant increase in the rate of TNF but not G3PDHmRNA degradation in the presence of the transcription inhibitor, actinomycin D. The decay of TNFmRNA is accelerated 1.7-fold, and its half-life is decreased by 57%. In view of its chemical and physical properties, we propose that 15-HPETE may function by destabilizing TNFmRNA by interaction with a trans-activating protein bound to the AU-rich element of TNFmRNA. PMID- 15749846 TI - Spontaneously Ig-secreting B-1 cells violate the accepted paradigm for expression of differentiation-associated transcription factors. AB - B-1 cells spontaneously secrete natural Ig that acts as a primary line of defense against infection. A major shortfall in our understanding of this key process centers on the molecular mechanisms regulating natural Ab secretion by B-1 cells. Herein, we demonstrate that secreting B-1 cells use some aspects of the recently recognized plasmacytic differentiation program but deviate from it in important ways. Specifically, we show that key repressors of the plasmacytic program, B cell leukemia/lymphoma-6 and paired box gene 5, are reduced in spontaneously secreting B-1 B cells, as in stimulated differentiated B-2 cells. Surprisingly, we find that key promoters of the plasmacytic program, B lymphocyte inducer of maturation program 1 and X-box binding protein 1, are not up-regulated in secreting B-1 cells, in contrast to secreting B-2 cells. These data demonstrate that B-1 cells operate under a differentiation program that is unique and differs from the paradigm associated with Ig-secreting B-2 cells. PMID- 15749847 TI - The mouse CD1d cytoplasmic tail mediates CD1d trafficking and antigen presentation by adaptor protein 3-dependent and -independent mechanisms. AB - The short cytoplasmic tail of mouse CD1d (mCD1d) is required for its endosomal localization, for the presentation of some glycolipid Ags, and for the development of Valpha14i NKT cells. This tail has a four-amino acid Tyr containing motif, Tyr-Gln-Asp-Ile (YQDI), similar to those sequences known to be important for the interaction with adaptor protein complexes (AP) that mediate the endosomal localization of many different proteins. In fact, mCD1d has been shown previously to interact with the AP-3 adaptor complex. In the present study, we mutated each amino acid in the YQDI motif to determine the importance of the entire motif sequence in influencing mCD1d trafficking, its interaction with adaptors, and its intracellular localization. The results indicate that the Y, D, and I amino acids are significant functionally because mutations at each of these positions altered the intracellular distribution of mCD1d and reduced its ability to present glycosphingolipids to NKT cells. However, the three amino acids are not all acting in the same way because they differ with regard to how they influence the intracellular distribution of CD1d, its rate of internalization, and its ability to interact with the mu subunit of AP-3. Our results emphasize that multiple steps, including interactions with the adaptors AP-2 and AP-3, are required for normal trafficking of mCD1d and that these different steps are mediated by only a few cytoplasmic amino acids. PMID- 15749848 TI - Rationally engineered therapeutic proteins with reduced immunogenicity. AB - Chronic administration of protein therapeutics may elicit unacceptable immune responses to the specific protein. Our hypothesis is that the immunogenicity of protein drugs can be ascribed to a few immunodominant helper T lymphocyte (HTL) epitopes, and that reducing the MHC binding affinity of these HTL epitopes contained within these proteins can generate drugs with lower immunogenicity. To test this hypothesis, we studied the protein therapeutic erythropoietin (Epo). Two regions within Epo, designated Epo 91-120 and Epo 126-155, contained HTL epitopes that were recognized by individuals with numerous HLA-DR types, a property common to immunodominant HTL epitopes. We then engineered analog epitopes with reduced HLA binding affinity. These analog epitopes were associated with reduced in vitro immunogenicity. Two modified forms of Epo containing these substitutions were shown to be bioactive and nonimmunogenic in vitro. These findings support our hypothesis and demonstrate that immunogenicity of protein drugs can be reduced in a systematic and predictable manner. PMID- 15749849 TI - CD4- plasmacytoid dendritic cells (pDCs) migrate in lymph nodes by CpG inoculation and represent a potent functional subset of pDCs. AB - We have recently identified two groups of plasmacytoid dendritic cells (pDCs) isolated from murine liver based on the expression of CD4 and other cell surface markers uniquely expressed by pDCs. Herein, we describe the identification of both CD4+ and CD4- pDCs that clearly exist in lymph nodes (LNs), spleen, liver, thymus, bone marrow, and lung. Normally, CD4+ pDCs are enriched in LNs. However, after in vivo systemic injection with bacterial CpG, a larger number of CD4- pDCs are recruited to the LNs and local inoculation by CpG drives CD4- pDCs migrating into local sentinel LNs, suggesting that CD4- pDCs are the main subpopulation migrating to the peripheral LNs. Furthermore, although both freshly isolated CD4+ pDCs and CD4- pDCs appear as an immature plasmacytoid cell and develop into a DC morphology following activation, the two subsets have strikingly different immune features, including differences in the production pattern of cytokines stimulated with CpG and in T cell activation. PMID- 15749850 TI - Clustering of T cell ligands on artificial APC membranes influences T cell activation and protein kinase C theta translocation to the T cell plasma membrane. AB - T cell activation is associated with active clustering of relevant molecules in membrane microdomains defined as the supramolecular activation cluster. The contact area between these regions on the surface of T cells and APC is defined as the immunological synapse. It has been recently shown that preclustering of MHC-peptide complexes in membrane microdomains on the APC surface affects the efficiency of immune synapse formation and the related T cell activation. Disruption of such clusters may reduce the efficiency of stimulation. We describe here an entirely artificial system for Ag-specific, ex vivo stimulation of human polyclonal T cells (artificial APC (aAPC)). aAPC are based on artificial membrane bilayers containing discrete membrane microdomains encompassing T cell ligands (i.e., appropriate MHC-peptide complexes in association with costimulatory molecules). We show here that preclustering of T cell ligands triggered a degree of T cell activation significantly higher than the one achieved when we used either soluble tetramers or aAPC in which MHC-peptide complexes were uniformly distributed within artificial bilayer membranes. This increased efficiency in stimulation was mirrored by increased translocation from the cytoplasm to the membrane of protein kinase theta, a T cell signaling molecule that colocalizes with the TCR within the supramolecular activation cluster, thus indicating efficient engagement of T cell activation pathways. Engineered aAPC may have immediate application for basic and clinical immunology studies pertaining to modulation of T cells ex vivo. PMID- 15749851 TI - Impaired cytolytic activity in calreticulin-deficient CTLs. AB - Calreticulin is an endoplasmic reticulum-resident chaperone that is stored in the cytotoxic granules of CTLs and NK cells and is released with granzymes and perforin upon recognition of target cells. To investigate the role of calreticulin in CTL-mediated killing, we generated CTL lines from crt(+/+) and crt(-/-) mice expressing a constitutively active form of calcineurin in the heart. Crt(-/-) CTLs showed reduced cytotoxic activity toward allogeneic target cells despite normal production, intracellular localization, and activity of granzymes and despite perforin overexpression. Comparable or higher amounts of granzymes were degranulated by crt(-/-) cells in response to immobilized anti-CD3 Abs, indicating that calreticulin is dispensable for the signal transduction that leads to granule exocytosis. The ability to form conjugates with target cells was affected in the crt(-/-) CTLs, explaining the observed reduction in cytotoxicity. Conjugate formation and cytotoxicity were completely restored by treatments that facilitate recognition and contact with target cells, a prerequisite for degranulation and killing. Therefore, we conclude that calreticulin is dispensable for the cytolytic activity of granzymes and perforin, but it is required for efficient CTL-target cell interaction and for the formation of the death synapse. PMID- 15749852 TI - Mannose-binding lectin-deficient mice display defective apoptotic cell clearance but no autoimmune phenotype. AB - Mannose-binding lectin (MBL) is a circulating serum protein that is sequestered to sites of inflammation and infection. MBL is a member of the collectin family with structural similarities to the lung collectins and functional similarities to C1q. Both MBL and C1q activate complement; C1q activates the classical pathway and MBL the lectin pathway. Here we demonstrate that MBL binds apoptotic cells in vitro and confirm a role for MBL in clearance of apoptotic cells in vivo. Despite MBL null mice demonstrating defective apoptotic cell clearance they did not develop spontaneous autoimmunity, lymphoproliferation, or germinal center expansion although increased numbers of peritoneal B1 cells were detected. These data demonstrate an important in vivo role for MBL in clearance of dying cells and adds the MBL null animals to the few animals with demonstrable in vivo apoptotic cell clearance defects. Moreover, it demonstrates that failure of apoptotic cell clearance can be dissociated from autoimmunity. PMID- 15749854 TI - Probiotics ameliorate recurrent Th1-mediated murine colitis by inducing IL-10 and IL-10-dependent TGF-beta-bearing regulatory cells. AB - Recent studies of murine models of mucosal inflammation suggest that, whereas some kinds of bacterial microflora are inducers of disease, others, known as probiotics, prevent disease. In the present study, we analyzed the regulatory cytokine and cell response to probiotic (VSL#3) administration in the context of the Th1 T cell colitis induced by trinitrobenzene sulfonic acid treatment of SJL/J mice. Daily administration of probiotics for 3 wk to mice during a remission period between a first and second course of colitis induced by trinitrobenzene sulfonic acid, resulted in a milder form of recurrent colitis than observed in mice administered PBS during this same period. This protective effect was attributable to effects on the lamina propria mononuclear cell (LPMC) population, because it could be transferred by LPMC from probiotic-treated mice to naive mice. Probiotic administration was associated with an early increase in the production of IL-10 and an increased number of regulatory CD4+ T cells bearing surface TGF-beta in the form of latency-associated protein (LAP) (LAP+ T cells). The latter were dependent on the IL-10 production because administration of anti-IL-10R mAb blocked their appearance. Finally, the LAP+ T cells were essential to the protective effect of probiotics because administration of anti IL-10R or anti-TGF-beta at the initiation of recurrent colitis induction or depletion of LAP+ T cells from LPMC abolished the latter's capacity to transfer protection to naive recipients. These studies show that probiotic (VSL#3) administration during a remission period ameliorates the severity of recurrent colitis by inducing an immunoregulatory response involving TGF-beta-bearing regulatory cells. PMID- 15749853 TI - Heat shock protein 60 inhibits Th1-mediated hepatitis model via innate regulation of Th1/Th2 transcription factors and cytokines. AB - Extracellular heat shock protein 60 (HSP60) has been considered a proinflammatory danger signal. Yet, HSP60 can also down-regulate experimental immune arthritis and diabetes models by specific inhibition of Th1-like responses. We now report that HSP60 in vitro differentially modulates the expression of Th1/Th2 transcription factors in human T cells: HSP60 down-regulates T-bet, NF-kappaB, and NFATp and up-regulates GATA-3, leading to decreased secretion of TNF-alpha and IFN-gamma and enhanced secretion of IL-10. These effects depended on TLR2 signaling and could not be attributed to LPS or to other contaminants. In BALB/c mice, HSP60 in vivo inhibited the clinical, histological, and serological manifestations of Con A-induced hepatitis associated with up-regulated T cell expression of suppressor of cytokine signaling 3 and GATA-3 and down-regulated T bet expression. These results provide a molecular explanation for the effects of HSP60 treatment on T cell inflammation via innate regulation of the inflammatory response. PMID- 15749855 TI - Very low-dose tolerance with nucleosomal peptides controls lupus and induces potent regulatory T cell subsets. AB - We induced very low-dose tolerance by injecting lupus prone (SWR x NZB)F1 (SNF1) mice with 1 mug nucleosomal histone peptide autoepitopes s.c. every 2 wk. The subnanomolar peptide therapy diminished autoantibody levels and prolonged life span by delaying nephritis, especially by reducing inflammatory cell reaction and infiltration in kidneys. H4(71-94) was the most effective autoepitope. Low-dose tolerance therapy induced CD8+, as well as CD4+ CD25+ regulatory T (Treg) cell subsets containing autoantigen-specific cells. These adaptive Treg cells suppressed IFN-gamma responses of pathogenic lupus T cells to nucleosomal epitopes at up to a 1:100 ratio and reduced autoantibody production up to 90-100% by inhibiting nucleosome-stimulated T cell help to nuclear autoantigen-specific B cells. Both CD4+ CD25+ and CD8+ Treg cells produced and required TGF-beta1 for immunosuppression, and were effective in suppressing lupus autoimmunity upon adoptive transfer in vivo. The CD4+ CD25+ T cells were partially cell contact dependent, but CD8+ T cells were contact independent. Thus, low-dose tolerance with highly conserved histone autoepitopes repairs a regulatory defect in systemic lupus erythematosus by generating long-lasting, TGF-beta-producing Treg cells, without causing allergic/anaphylactic reactions or generalized immunosuppression. PMID- 15749857 TI - Coligation of the B cell receptor with complement receptor type 2 (CR2/CD21) using its natural ligand C3dg: activation without engagement of an inhibitory signaling pathway. AB - C3dg is a cleavage product of the C3 component of complement that can facilitate the coligation of the complement receptor 2 (CR2/CD21) with the BCR via C3dg/Ag complexes. This interaction can greatly amplify BCR-mediated signaling events and acts to lower the threshold for B cell activation. Although previous studies have used anti-CR2 Abs or used chimeric Ags in the context of BCR transgenic mice as surrogate C3d-containing ligands, we have used a physiological form of C3d to study signaling in B cells from wild-type C57BL/6 mice. We find that while CR2 enhanced BCR signaling causes intracellular Ca2+ mobilization and total pTyr phosphorylation of an intensity comparable to optimal BCR ligation using anti-IgM Abs, it does so with limited activation of inhibitory effectors (such as CD22, Src homology region 2 domain containing phosphatase 1, and SHIP-1) and without substantial receptor cross-linking. In summary, we demonstrate that CR2-enhanced BCR signaling may proceed not only through the previously described amplification of positive signaling pathways, but is potentially augmented by a lack of normal inhibitory/feedback signaling. PMID- 15749856 TI - Protein conformation significantly influences immune responses to prion protein. AB - In prion diseases, such as variant Creutzfeldt-Jakob disease normal cellular prion protein (PrPC), a largely alpha-helical structure is converted to an abnormal conformational isoform (PrPSc) that shows an increase in beta-sheet content. Similarly, the recombinant form of PrPC (ralpha-PrP) can be converted to a conformation dominated by beta-sheet (rbeta-PrP) by reduction and mild acidification in vitro, a process that may mimic in vivo conversion following PrPC internalization during recycling. Despite PrPSc accumulation and prion propagation in the lymphoreticular system before detectable neuroinvasion, no Ab response to PrP has been detected, probably due to immune tolerance. To investigate how the immune system may respond to alpha- and beta-PrP, we immunized Prnp(0/0) mice that are not tolerant of PrP with ralpha-PrP and rbeta PrP. In this study, we show that although T cells stimulated by these differently folded conformers PrP recognize similar immunodominant epitopes (residues 111-130 and 191-210) the cytokine profile in response to ralpha- and rbeta-PrP was different. Challenge with ralpha-PrP elicited a strong response of IL-5 and IL 10, whereas rbeta-PrP led to an early increased production of IFN-gamma. In addition, immunization with ralpha-PrP led to production of predominantly IgG1 isotype Ab in the sera, whereas after immunization with rbeta-PrP, IgG2b was significantly produced. Thus, both humoral and cellular responses to these differently folded isoforms of the same protein are different, indicating a possible involvement of Th1 and Th2 pathway activation. These differences may be exploitable diagnostically and therapeutically for prion diseases, such as variant Creutzfeldt-Jakob disease. PMID- 15749858 TI - Exacerbation of granuloma formation in IL-1 receptor antagonist-deficient mice with impaired dendritic cell maturation associated with Th2 cytokine production. AB - Dendritic cell (DC) maturation at the site of inflammation and migration into draining lymph nodes is fundamental to initiate Ag-specific immune responses. Although several proinflammatory cytokines, including IL-1, are known to promote DC maturation in vitro, their contributions to DC activation and migration within peripheral inflamed tissue compartments are not yet fully understood. We show here that endogenous IL-1 receptor antagonist (IL-1ra) controls the activation state of liver-recruited DCs and their migration in a Propionibacterium acnes induced murine granulomatous liver disease model. After P. acnes treatment, formation of portal tract-associated lymphoid tissue was conversely impaired in IL-1ra-deficient mice. IL-1ra-deficient mice developed hepatic granulomas within 3 days after P. acnes administration and showed a more pronounced granuloma formation than wild-type mice. Although sinusoidal granulomas contained numerous CD11c+ DCs at day 7, expressions of CCR7, IL-12p40 by these DCs were dramatically decreased in IL-1ra-deficient mice, suggesting aberrant DC maturation and sinusoid portal migration in the absence of endogenous IL-1ra. This was accompanied with enhanced intrahepatic Th2 cytokine production and severe hepatocellular damage. Thus, hepatocyte-derived IL-1ra may control optimal activation and migration of inflammatory DCs within the liver and thereby determine the local immune responses in granulomatous liver disease. PMID- 15749859 TI - Delta-9-tetrahydrocannabinol enhances breast cancer growth and metastasis by suppression of the antitumor immune response. AB - In the current study, we tested the central hypothesis that exposure to Delta-9 tetrahydrocannabinol (Delta9-THC), the major psychoactive component in marijuana, can lead to enhanced growth of tumors that express low to undetectable levels of cannabinoid receptors by specifically suppressing the antitumor immune response. We demonstrated that the human breast cancer cell lines MCF-7 and MDA-MB-231 and the mouse mammary carcinoma 4T1 express low to undetectable levels of cannabinoid receptors, CB1 and CB2, and that these cells are resistant to Delta9-THC-induced cytotoxicity. Furthermore, exposure of mice to Delta9-THC led to significantly elevated 4T1 tumor growth and metastasis due to inhibition of the specific antitumor immune response in vivo. The suppression of the antitumor immune response was mediated primarily through CB2 as opposed to CB1. Furthermore, exposure to Delta9-THC led to increased production of IL-4 and IL-10, suggesting that Delta9-THC exposure may specifically suppress the cell-mediated Th1 response by enhancing Th2-associated cytokines. This possibility was further supported by microarray data demonstrating the up-regulation of a number of Th2-related genes and the down-regulation of a number of Th1-related genes following exposure to Delta9-THC. Finally, injection of anti-IL-4 and anti-IL-10 mAbs led to a partial reversal of the Delta9-THC-induced suppression of the immune response to 4T1. Such findings suggest that marijuana exposure either recreationally or medicinally may increase the susceptibility to and/or incidence of breast cancer as well as other cancers that do not express cannabinoid receptors and are resistant to Delta9-THC-induced apoptosis. PMID- 15749860 TI - The generation of CD25+ CD4+ regulatory T cells that prevent allograft rejection does not compromise immunity to a viral pathogen. AB - In all but a small minority of cases, continued survival of solid organ grafts after transplantation depends on lifelong, nonselective immunosuppression that, although effective, results in increased rates of infection, cancer, and vascular disease. Therapeutic strategies that engage or mimic self-tolerance may allow prolonged allograft survival without the disadvantages of nonspecific immunotherapy. Pretreatment of recipient mice with donor alloantigen combined with transient modulation of the peripheral T cell pool with anti-CD4 Ab leads to the indefinite survival of MHC-incompatible cardiac allografts without further therapy. Tolerance is dependent on CD25+ CD4+ regulatory T cells that arise from naive CD25- precursors and regulate rejection via both IL-10 and CTLA-4. Although these cells are clearly effective at controlling rejection, the proven ability of recently activated CD25+ cells to mediate bystander regulation raises the possibility that tolerized individuals might also have a reduced capacity to respond to environmental pathogens. We have examined anti-influenza responses in tolerized primary heart recipients, secondary recipients following adoptive transfer of regulatory populations, and tolerized mice in which bystander regulation has been deliberately induced. Neither virus-specific CTL activity in vitro nor the clearance of virus in vivo was significantly diminished in any of these treatment groups compared with infected unmanipulated controls. The data suggest that the induction of dominant allograft tolerance dependent on regulatory T cells does not necessarily result in attenuated responses to pathogens providing further support for the development of tolerance induction protocols in clinical transplantation. PMID- 15749861 TI - CD38 controls ADP-ribosyltransferase-2-catalyzed ADP-ribosylation of T cell surface proteins. AB - ADP-ribosyltransferase-2 (ART2), a GPI-anchored, toxin-related ADP-ribosylating ectoenzyme, is prominently expressed by murine T cells but not by B cells. Upon exposure of T cells to NAD, the substrate for ADP-ribosylation, ART2 catalyzes ADP-ribosylation of the P2X7 purinoceptor and other functionally important cell surface proteins. This in turn activates P2X7 and induces exposure of phosphatidylserine and shedding of CD62L. CD38, a potent ecto-NAD-glycohydrolase, is strongly expressed by most B cells but only weakly by T cells. Following incubation with NAD, CD38-deficient splenocytes exhibited lower NAD glycohydrolase activity and stronger ADP-ribosylation of cell surface proteins than their wild-type counterparts. Depletion of CD38(high) cells from wild-type splenocytes resulted in stronger ADP-ribosylation on the remaining cells. Similarly, treatment of total splenocytes with the CD38 inhibitor nicotinamide 2' deoxy-2'-fluoroarabinoside adenine dinucleotide increased the level of cell surface ADP-ribosylation. Furthermore, the majority of T cells isolated from CD38 deficient mice "spontaneously" exposed phosphatidylserine and lacked CD62L, most likely reflecting previous encounter with ecto-NAD. Our findings support the notion that ecto-NAD functions as a signaling molecule following its release from cells by lytic or nonlytic mechanisms. ART2 can sense and translate the local concentration of ecto-NAD into corresponding levels of ADP-ribosylated cell surface proteins, whereas CD38 controls the level of cell surface protein ADP ribosylation by limiting the substrate availability for ART2. PMID- 15749862 TI - Identification of stimulating and inhibitory epitopes within the heat shock protein 70 molecule that modulate cytokine production and maturation of dendritic cells. AB - The 70-kDa microbial heat shock protein (mHSP70) has a profound effect on the immune system, interacting with the CD40 receptor on DC and monocytes to produce cytokines and chemokines. The mHSP70 also induces maturation of dendritic cells (DC) and thus acts as an alternative ligand to CD40L on T cells. In this investigation, we have identified a cytokine-stimulating epitope (peptide 407 426), by activating DC with overlapping synthetic peptides (20-mers) derived from the sequence of mHSP70. This peptide also significantly enhances maturation of DC stimulated by mHSP70 or CD40L. The epitope is located at the base of the peptide binding groove of HSP70 and has five critical residues. Furthermore, an inhibitory epitope (p457-496) was identified downstream from the peptide-binding groove that inhibits cytokine production and maturation of DC stimulated by HSP70 or CD40L. The p38 MAP kinase phosphorylation is critical in the alternative CD40 HSP70 pathway and is inhibited by p457-496 but enhanced by p407-426. PMID- 15749863 TI - The immunogenicity of dendritic cell-based vaccines is not hampered by doxorubicin and melphalan administration. AB - Immunization of cancer patients is most effective in tumor-free conditions or in the presence of minimal residual disease. In the attempt to develop new strategies able to control tumor recurrence while allowing the development of protective immunity, we have investigated the immunogenic potential of two distinct vaccine formulations when provided alone or upon single and repeated treatment with chemotherapeutics drugs. Vaccine-induced T cell responses were first investigated by tracing Ag-specific T cell responses in mice bearing detectable frequencies of Ag-specific TCR transgenic CD4 and CD8 T cells. These studies indicated that immunization with peptide-pulsed dendritic cells and soluble Ag plus adjuvant elicited a comparable expansion and differentiation of CD4 and CD8 effector cells in the peripheral lymphoid tissues when provided alone or shortly after Doxorubicin or Melphalan administration. We also analyzed the potency of the combined vaccination in transgenic adenocarcinoma mouse prostate mice, which develop spontaneous prostate cancer. Dendritic cell-based vaccination elicited potent tumor-specific cytotoxic responses in mice bearing prostate intraepithelial neoplasia both in the absence and in the presence of Doxorubicin. Together our results indicate that Doxorubicin- or Melphalan-based chemotherapy and Ag-specific vaccination can be combined for adjuvant treatments of cancer patients. PMID- 15749864 TI - Nucleosome, the main autoantigen in systemic lupus erythematosus, induces direct dendritic cell activation via a MyD88-independent pathway: consequences on inflammation. AB - Nucleosome is the major autoantigen in systemic lupus erythematosus. It is found as a circulating complex in the sera of patients and seems to play a key role in disease development. In this study, we show for the first time that physiologic concentrations of purified nucleosomes directly induce in vitro dendritic cell (DC) maturation of mouse bone marrow-derived DC, human monocyte-derived DC (MDDC), and purified human myeloid DC as observed by stimulation of allogenic cells in MLR, cytokine secretion, and CD86 up-regulation. Importantly, nucleosomes act as free complexes without the need for immune complex formation or for the presence of unmethylated CpG DNA motifs, and we thus identified a new mechanism of DC activation by nucleosomes. We have clearly demonstrated that this activation is nucleosome-specific and endotoxin-independent. Particularly, nucleosomes induce MDDC to secrete cytokines known to be detected in high concentrations in the sera of patients. Moreover, activated MDDC secrete IL-8, a neutrophil chemoattractant also detected in patient sera, and thus might favor the inflammation observed in patients. Both normal and lupus MDDC are sensitive to nucleosome-induced activation. Finally, injection of purified nucleosomes to normal mice induces in vivo DC maturation. Altogether, these results strengthen the key role of nucleosomes in systemic lupus erythematosus and might explain how peripheral tolerance is broken in patients. PMID- 15749865 TI - Mechanisms that limit the in vitro proliferative potential of human CD8+ T lymphocytes. AB - Human T lymphocytes can be numerically expanded in vitro only to a limited extent. The cyclin-dependent kinase inhibitor p16(INK4a) is essential in the control of cellular proliferation, and its expression, in epithelial cells, is associated with irreversible growth arrest. Using long-term cultured CD8+ T lymphocytes, we have investigated the role of the p16/pRb pathway in the regulation of T cell proliferation and senescence. In this study, we describe at least two mechanisms that cause replicative growth arrest in cultured lymphocytes. The first one depends on the expression of p16(INK4a) and is directly responsible for the exit of a significant proportion of CD8+ T cells from the proliferative population. This induced p16 expression pattern is observed during each round of mitogen stimulation and is not related to activation-induced cell death. Importantly, knocking down p16(INK4a) expression allows increased proliferation of T cells. The second one is a phenomenon that resembles human fibroblast senescence, but is independent of p16(INK4a) and of telomere attrition. Interestingly, virtually all pRb proteins in the senescent population are found in the active form. Our data indicate that newly synthesized p16(INK4a) limits the proliferation of T lymphocytes that respond to mitogen, but is not required for the loss of mitogen responsiveness called senescence. PMID- 15749866 TI - Regulatory T cells suppress CD8+ T cell responses induced by direct priming and cross-priming and moderate immunodominance disparities. AB - Little is known regarding the participation of CD4+ CD25+ regulatory T cells (Treg) in TCD8+ responses. In this study, we show that Treg depletion via treatment with anti-CD25 mAb (PC61) significantly enhances TCD8+ responses to influenza A virus, vaccinia virus, and SV40-transformed cells induced by either direct priming or cross-priming. PC61 did not enhance TCD8+ responses in CD4 deficient mice, providing the initial demonstration that PC61 acts on a subset of TCD4+, and not on other cells that express either CD25 or a fortuitously cross reactive Ag. We further show that Treg selectively suppress responses to the most immunodominant TCD8+ determinants in the three systems examined. Therefore, Treg influence TCD8 immunodominance hierarchies by moderating disparities in responses to different determinants. PMID- 15749867 TI - Amelioration of established experimental autoimmune encephalomyelitis by an MHC anchor-substituted variant of proteolipid protein 139-151. AB - Murine experimental autoimmune encephalomyelitis (EAE) is a CD4+ T cell-mediated autoimmune disorder directed against myelin proteins within the CNS. We propose that variant peptides containing amino acid substitutions at MHC anchor residues will provide a unique means to controlling the polyclonal autoimmune T cell response. In this study, we have identified an MHC variant of proteolipid protein (PLP) 139-151 (145D) that renders PLP(139-151)-specific T cell lines anergic in vitro, as defined by a significant reduction in proliferation and IL-2 production following challenge with wild-type peptide. In vivo administration of 145D before challenge with PLP(139-151) results in a significant reduction in disease severity and incidence. Importantly, we demonstrate the ability of an MHC variant peptide to ameliorate established EAE. An advantage to this treatment is that the MHC variant peptide does not induce an acute hypersensitivity reaction. This is in contrast to previous work in the PLP(139-151) model demonstrating that anaphylactic shock resulting in death occurs upon rechallenge with the encephalitogenic peptide. Taken together, these data demonstrate the effectiveness of MHC anchor-substituted peptides in the treatment of EAE and suggest their utility in the treatment of other autoimmune disorders. PMID- 15749868 TI - Opposing roles for the cyclin-dependent kinase inhibitor p27kip1 in the control of CD4+ T cell proliferation and effector function. AB - Cell division drives T cell clonal expansion and differentiation, and is the result of concerted signaling from Ag, costimulatory, and growth factor receptors. How these mitogenic signals are coupled to the cell cycle machinery in primary T cells is not clear. We have focused on the role of p27kip1, a major cyclin-dependent kinase binding protein expressed by CD4+ T cells. Our studies using p27kip1 gene dosage demonstrate that early after activation, p27kip1 acts to promote, rather than inhibit, G1 to S phase progression within the first division cycle. However, throughout subsequent cell divisions p27kip1 behaves as a negative regulator, directly establishing the threshold amount of growth factor signaling required to support continued cell division. During this phase, signals from CD28 and IL-2R cooperate with the TCR to "tune" this threshold by inducing the degradation of p27kip1 protein, and we show that agents that block these pathways require elevated p27kip1 levels for their full antiproliferative activity. Finally, we show that p27kip1 opposes the development of CD4+ T cell effector function, and is required for the full development of anergy in response to a tolerizing stimulus. Our results suggest that p27kip1 plays a complex and important role in the regulation of cell division and effector function in primary CD4+ T cells. PMID- 15749869 TI - B cell receptor (BCR) cross-talk: CD40 engagement enhances BCR-induced ERK activation. AB - Bystander B cells may be initially stimulated through CD40, which enhances susceptibility to Fas-mediated apoptosis, before encountering Ag, which produces Fas resistance. A key issue in this process is to what extent CD40 cross-talk might affect subsequent BCR signaling. It has previously been shown that CD40 engagement bypasses or mitigates the need for Bruton's tyrosine kinase in subsequent BCR signaling for NF-kappaB activation. However, the full extent of the effects of CD40 on BCR signaling has not been delineated. In the present study we evaluated the possibility that CD40-mediated cross-talk also affects another principal outcome of BCR signaling: MAPK activation. We found that prior stimulation of primary murine B cells with CD40L markedly enhanced the level of ERK and JNK (but not p38 MAPK) phosphorylation produced by subsequently added anti-Ig Ab, and much, but not all, of this enhancement was independent of PI3K and phospholipase C. CD40L treatment similarly enhanced BCR-induced MAPK kinase (MEK) phosphorylation, and MEK was required for enhancement of ERK. Although BCR induced c-Raf phosphorylation was also enhanced by prior CD40L treatment, c-Raf was not required for MEK/ERK phosphorylation. These results identify a novel system of receptor cross-talk between CD40 and BCR and indicate that the effects of CD40 engagement on subsequent BCR stimulation spread beyond NF-kappaB to involve the MAPK pathway. PMID- 15749870 TI - B and T lymphocyte attenuator exhibits structural and expression polymorphisms and is highly Induced in anergic CD4+ T cells. AB - B and T lymphocyte attenuator (BTLA) was initially identified as expressed on Th1 cells and B cells, but recently reported to be expressed by macrophages, dendritic cells, and NK cells as well. To address this discrepancy we generated a panel of BTLA-specific mAbs and characterized BTLA expression under various activation conditions. We report the existence of three distinct BTLA alleles among 23 murine strains, differing both in Ig domain structure and cellular distribution of expression on lymphoid subsets. The BALB/c and MRL/lpr alleles differ at one amino acid residue, but C57BL/6 has nine additional differences and alters the predicted cysteine bonding pattern. The BALB/c BTLA allele is also expressed by B cells, T cells, and dendritic cells, but not macrophages or NK cells. However, C57BL/6 BTLA is expressed on CD11b+ macrophages and NK cells. Finally, in CD4+ T cells, BTLA is expressed most highly following Ag-specific induction of anergy in vivo, and unlike programmed death-1 and CTLA-4, not expressed by CD25+ regulatory T cells. These results clarify discrepancies regarding BTLA expression, suggest that structural and expression polymorphisms be considered when analyzing BTLA in various murine backgrounds, and indicate a possible role in anergic CD4+ T cells. PMID- 15749871 TI - Gammadelta T cell function varies with the expressed WC1 coreceptor. AB - WC1 molecules are transmembrane glycoproteins belonging to the scavenger receptor cysteine-rich family and uniquely expressed on gammadelta T cells. Although participation of WC1+ gammadelta T cells in immune responses is well established, very little is understood regarding the significance of expressing different forms of the WC1 molecule. Two forms previously identified by mAbs, i.e., WC1.1 and WC1.2, are expressed by largely nonoverlapping subpopulations of gammadelta T cells. In this study it was shown that expression of the WC1.1 coreceptor was the main indicator of proliferation and IFN-gamma production in response to autologous and bacterial Ags as well as for IFN-gamma production without proliferation in Th1-polarizing, IL-12-containing cultures. Nevertheless, after culture in either Th1-polarizing or neutral conditions, mRNA was present for both T-bet and GATA-3 as well as for IL-12Rbeta2 in WC1.1+ and WC1.2+ subpopulations, and neither produced IL-4 under any conditions. Although the steady decrease in the proportion of WC1.1+ cells, but not WC1.2+ cells, within PBMC with animal aging suggested that the two subpopulations may have different roles in immune regulation, cells bearing either WC1.1 or WC1.2 expressed mRNA for regulatory cytokines IL-10 and TGF-beta, with TGF-beta being constitutively expressed by ex vivo cells. Overall, the results demonstrate that the form of the WC1 coreceptor expressed on gammadelta T cells divides them into functional subsets according to IFN-gamma production and proliferative capacity to specific stimuli as well as with regard to representation within PBMC. Finally, evidence is provided for minor differences in the intracytoplasmic tail sequences of WC1.1 and WC1.2 that may affect signaling. PMID- 15749872 TI - Small rho GTPases regulate antigen presentation in dendritic cells. AB - Dendritic cells (DC) are involved in the regulation of innate and adaptive immunity. However, the molecular mechanisms maintaining DC function remain to be elucidated. In this study, we report on the role of small Rho GTPases: Cdc42, Rac1, and RhoA in the regulation of DC adherence, Ag presentation, migration, chemotaxis, and endocytosis. Murine DC were transfected with vaccinia virus-based constructs, encoding dominant-negative or constitutively active (ca) mutant forms of Rho GTPases. We demonstrate that Cdc42 plays a major role in the regulation of DC adhesion, because caCdc42-transfected DC had significant up-regulation of adhesion to extracellular matrix, which was blocked by the Rho GTPase inhibitor toxin B (ToxB). In contrast, caRho-transfected DC only modestly elevated DC adhesion, and caRac had no effect. Additionally, caCdc42 and caRho increased the ability of DC to present OVA peptide to specific T cells. This effect was abrogated by ToxB. Activation of Cdc42 in DC significantly inhibited spontaneous and chemokine-induced DC migration. Furthermore, uptake of dextran 40 by DC was significantly enhanced by Rho GTPase activators cytotoxic necrotizing factor 1 and PMA, and reduced by ToxB. caCdc42 also increased endocytotic activity of DC, whereas dominant-negative Cdc42 blocked it. Thus, Rho GTPases Cdc42, RhoA, and Rac1 regulate DC functions that are critical for DC-mediated immune responses in vivo. PMID- 15749873 TI - The role of intercellular adhesion molecule-1/LFA-1 interactions in the generation of tumor-specific CD8+ T cell responses. AB - The activation of naive CD4+ T cells requires both TCR engagement and a second costimulatory signal mediated by the interaction of CD28 with CD80/CD86 expressed on professional APC. However, the situation for naive CD8+ T cells is less clear. Although evidence indicates that induction of CD8+ T cell responses is also dependent on professional APC, the ability of some tumors, which do not express CD80/CD86, to induce CTL suggests that other pathways of costimulation exist for the activation of CD8+ T cells. We examined the ability of tumor cells expressing different levels of a tumor-specific Ag to directly prime CD8+ T cells. We demonstrate that CD8+ T cells are directly activated by tumor cells in a CD80/CD86-CD28 independent manner. In this system, costimulation requires ICAM 1/LFA-1 interaction. This results in the generation of CTL capable of inhibiting tumor growth in vivo, and maintaining long-term survival. PMID- 15749875 TI - ICAM-1 contributes to but is not essential for tumor antigen cross-priming and CD8+ T cell-mediated tumor rejection in vivo. AB - ICAM-1 has been described to provide both adhesion and costimulatory functions during T cell activation. In the setting of antitumor immunity, ICAM-1/LFA-1 interactions could be important at the level of T cell priming by APCs in draining lymph nodes as well as for transendothelial migration and tumor cell recognition at the tumor site. To determine the contribution of ICAM-1 to tumor rejection in vivo, we performed adoptive transfer of 2C TCR-transgenic/RAG2(-/-) T cells into TCRalpha(-/-) vs ICAM(-/-)/TCRalpha(-/-) recipient animals. ICAM-1 deficient mice successfully rejected HTR.C tumors expressing Ld recognized by the 2C TCR, albeit with a kinetic delay. Inasmuch as HTR.C tumor cells themselves express ICAM-1, a second model was pursued using B16-F10 melanoma cells that lack ICAM-1 expression. These cells were transduced to express the SIYRYYGL peptide recognized by the 2C TCR in the context of Kb, which is cross-presented by APCs in H-2b mice in vivo. These tumors also grew more slowly but were eventually rejected by the majority of ICAM-1(-/-)/TCRalpha(-/-) recipients. Delayed rejection in ICAM-1(-/-) mice was associated with diminished T cell priming as assessed by ELISPOT. In contrast, T cell penetration into the tumor was comparable in wild-type and ICAM-1(-/-) hosts, and adoptively transferred primed effector 2C cells rejected normally in ICAM-1(-/-) recipients. Our results suggest that ICAM-1 contributes to but is not absolutely required for CD8+ T cell mediated tumor rejection in vivo and dominantly acts at the level of priming rather than the effector phase of the antitumor immune response. PMID- 15749874 TI - Role of the programmed death-1 pathway in regulation of alloimmune responses in vivo. AB - Programmed death-1 (PD-1), an inhibitory receptor up-regulated on activated T cells, has been shown to play a critical immunoregulatory role in peripheral tolerance, but its role in alloimmune responses is poorly understood. Using a novel alloreactive TCR-transgenic model system, we examined the functions of this pathway in the regulation of alloreactive CD4+ T cell responses in vivo. PD-L1, but not PD-1 or PD-L2, blockade accelerated MHC class II-mismatched skin graft (bm12 (I-Abm12) into B6 (I-Ab)) rejection in a similar manner to CTLA-4 blockade. In an adoptive transfer model system using the recently described anti-bm12 (ABM) TCR-transgenic mice directly reactive to I-Abm12, PD-1 and PD-L1 blockade enhanced T cell proliferation early in the immune response. In contrast, at a later time point preceding accelerated allograft rejection, only PD-L1 blockade enhanced T cell proliferation. In addition, PD-L1 blockade enhanced alloreactive Th1 cell differentiation. Apoptosis of alloantigen-specific T cells was inhibited significantly by PD-L1 but not PD-1 blockade, indicating that PD-1 may not be the receptor for the apoptotic effect of the PD-L1-signaling pathway. Interestingly, the effect of PD-L1 blockade was dependent on the presence of CD4+ CD25+ regulatory T cells in vivo. These data demonstrate a critical role for the PD-1 pathway, particularly PD-1/PD-L1 interactions, in the regulation of alloimmune responses in vivo. PMID- 15749876 TI - Quantitating effector and regulatory T lymphocytes in immune responses by limiting dilution analysis modeling. AB - Although there is currently no doubt that regulatory lymphocytes represent a master player in the immune system, a major unresolved problem is the accurate quantitation of these cells among unfractionated cell populations. This difficulty mainly arises because there are no specific immunophenotypic markers that can reliably discriminate between effector and regulatory lymphocytes. To face this problem, we have developed computational models of limiting dilution analyses addressing the question of the accurate estimation of the frequencies of effector and regulatory cells functionally engaged in an immune response. A set of generic equations were provided to form a framework for modeling limiting dilution data, enabling discrimination between qualitatively different models of suppression. These models include either one or two subpopulations of regulatory cells, featured by either low or potent regulatory activity. The potential of this modeling approach was illustrated by the accurate determination of the frequencies of effector and regulatory T lymphocytes in one real limiting dilution experiment of CD4+ CD25+ T lymphocytes performed in the context of an allogeneic response in the human system. The crucial advantage of the limiting dilution method over the "static, phenotype-based" method is the dynamic evaluation of effector and regulatory T cell biology through their actual functional activity. PMID- 15749877 TI - Cross-priming of T cell responses by synthetic microspheres carrying a CD8+ T cell epitope requires an adjuvant signal. AB - Controlling the cross-presentation of exogenous Ags to CD8+ T cells represents a major step for designing new vaccination strategies. Whereas several recombinant pseudo-viral particles have been used as delivery systems for triggering potent CTL responses to heterologous exogenous Ags, the adjuvant properties of virus like particles (VLPs) themselves were little questioned. Here, we analyzed the contribution of the porcine parvovirus (PPV)-VLPs to the induction of protective cellular responses to exogenous Ags carried by an independent delivery system. Microspheres, which are known to transfer exogenous Ags into the MHC class I pathway, were chosen for delivering the immunodominant OVA(257-264) CD8+ T cell epitope (B-OVAp). This delivery system fulfills the requirements in terms of cross-presentation, but fails to induce cross-priming of specific CD8+ T cells. Coinjection of PPV-VLPs with B-OVAp results in the priming of potent CTL responses and type 1-biased immunity in a CD4- and CD40-independent manner, as efficiently as the recombinant PPV-VLPs carrying the same epitope (PPV-OVAp). Furthermore, vaccination with PPV-VLPs and B-OVAp was fully efficient to protect mice against the development of OVA-bearing melanoma. These findings indicate that PPV-VLPs act not only as a delivery system but also as a strong adjuvant when independently provided with exogenous Ag. Thus, dissociation between delivery system and adjuvant would provide a more flexible and reliable system to induce potent and protective CTL. PMID- 15749878 TI - A role for caspases in controlling IL-4 expression in T cells. AB - Although caspase activation is critical for T cell proliferation following activation, the role of caspases in T cell differentiation is unclear. In this study, we have examined the effect of inhibition of caspases on the process of Th1/Th2 differentiation. Naive CD4+ T cells activated under neutral differentiation conditions in the presence of the pan caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp (Z-VAD) fluoromethylketone showed increased Th2 cell differentiation concomitant with an up-regulation of GATA-3. Z-VAD induced optimal Th2 differentiation when T cells were stimulated under strong primary activation conditions. Treatment of naive CD4+ T cells with Z-VAD under strong activation conditions led to a 6-fold increase in IL-4 mRNA compared with control treated T cells. The Z-VAD-induced increase in IL-4 transcription occurred within 24 h of activation and was independent of Stat6. IFN-gamma mRNA expression was not affected by Z-VAD at the 24-h time point. Z-VAD did not augment IL-4 expression from a committed Th2 cell, suggesting that caspases regulate IL-4 expression specifically during primary T cell activation. Z-VAD did not augment IL-12-driven Th1 differentiation. Activation of T cells in the presence of Z-VAD led to a specific increase in the expression of the transcription factor c-fos. Lastly, retrovirus-mediated expression of the antiapoptotic protein Bcl-2 resulted in an enhancement of Th2 cytokine expression, suggesting that inhibition of caspase activation by Bcl-2 can also modulate IL-4 expression. These findings reveal a novel regulatory mechanism of cytokine expression by caspases, and may explain how signaling pathways that inhibit apoptosis tend to promote Th2 differentiation. PMID- 15749879 TI - Transplantation tolerance in NF-kappaB-impaired mice is not due to regulation but is prevented by transgenic expression of Bcl-xL. AB - NF-kappaB is a key regulator of transcription after TCR and costimulatory receptor ligation. To determine the role of T cell-intrinsic NF-kappaB activation in acute allograft rejection, we used IkappaBalphaDeltaN-Tg mice (H-2b) that express an inhibitor of NF-kappaB restricted to the T cell compartment. We have previously shown that these mice permanently accept fully allogeneic (H-2d) cardiac grafts and secondary donor skin grafts, and that splenocytes from these tolerant mice have reduced alloreactivity when restimulated in vitro. These results were compatible with either deletion or suppression of allospecific T cells as possible mechanisms of tolerance. The aim of this study was to investigate the mechanism of transplant tolerance in these mice. IkappaBalphaDeltaN-Tg mice did not have increased numbers or function of CD4+ CD25+ regulatory T cells either before or after cardiac transplantation. In addition, tolerance could not be transferred to fresh NF-kappaB-competent T cells and was not permissive for linked suppression to skin grafts sharing donor and third-party alloantigens, suggesting that dominant suppression is not the mechanism by which IkappaBalphaDeltaN-Tg mice achieve tolerance. In contrast, overexpression of the antiapoptotic protein Bcl-xL in T cells from IkappaBalphaDeltaN-Tg mice resulted in effective rejection of cardiac allografts and correlated with an increased frequency of splenocytes producing IFN-gamma in response to alloantigen. Together, these results suggest that the death of alloreactive T cells may be partly responsible for the transplantation tolerance observed in mice with defective T cell-intrinsic NF-kappaB activation. PMID- 15749880 TI - VH1-46 is the dominant immunoglobulin heavy chain gene segment in rotavirus specific memory B cells expressing the intestinal homing receptor alpha4beta7. AB - Memory B cells expressing the intestinal homing marker alpha4beta7 are important for protective immunity against human rotavirus (RV). It is not known whether the B cell repertoire of intestinal homing B cells differs from B cells of the systemic compartment. In this study, we analyzed the RV-specific VH and VL repertoire in human IgD- B cells expressing the intestinal homing marker alpha4beta7. The mean frequency of RV-specific B cells in the systemic compartment of healthy adult subjects was 0.6% (range, 0.2-1.2). The mean frequency of IgD- B cells that were both RV specific and alpha4beta7 was 0.04% (range, 0.01-0.1), and a mean of 10% (range, 1-32) of RV-specific peripheral blood human B cells exhibited an intestinal homing phenotype. We previously demonstrated that VH1-46 is the dominant Ab H chain gene segment in RV-specific systemic B cells from adults and infants. RV-specific systemic IgD- or intestinal homing IgD-/alpha4beta7+ B cells in the current study also used the gene segment VH1-46 at a high frequency, while randomly selected B cells with those phenotypes did not. These data show that VH1-46 is the immunodominant gene segment in human RV-specific effector B cells in both the systemic compartment and in intestinal homing lymphocytes. The mean replacement/silent mutation ratio of systemic compartment IgD- B cells was >2, consistent with a memory phenotype and antigenic selection. Interestingly, RV-specific intestinal homing IgD-/alpha4beta7+ B cells using the VH1-46 gene segment were not mutated, in contrast to systemic RV specific IgD- B cells. PMID- 15749882 TI - Solvent accessibility of native and hydrolyzed human complement protein 3 analyzed by hydrogen/deuterium exchange and mass spectrometry. AB - Complement protein C3 is a 187-kDa (1641-aa) protein that plays a key role in complement activation and immune responses. Its hydrolyzed form, C3(H2O), is responsible for the initiation of the activation of alternative complement pathway. Previous analyses using mAbs, anilinonaphthalenesulfonate dyes, and functional studies have suggested that C3 is conformationally different from C3(H2O). We have used amide hydrogen/deuterium exchange and MALDI-TOF mass spectrometry to identify and localize structural differences between native C3 and C3(H2O). Both proteins were incubated in D2O for varying amounts of time, digested with pepsin, and then subjected to mass-spectrometric analysis. Of 111 C3 peptides identified in the MALDI-TOF analysis, 31 had well-resolved isotopic mass envelopes in both C3 and C3(H2O) spectra. Following the conversion of native C3 to C3(H2O), 17 of these 31 peptides exhibited a change in deuterium incorporation, suggesting a conformational change in these regions. Among the identified peptides, hydrogen/deuterium exchange data were obtained for peptides 944-967, 1211-1228, 1211-1231, 1259-1270, 1259-1273, 1295-1318, and 1319-1330, which span the factor H binding site on C3d and factor I cleavage sites, and peptides 1034-1048, 1049-1058, 1069-1080, 1130-1143, 1130-1145, 1211-1228, 1211 1231, 1259-1270, and 1259-1273, spanning 30% of the C3d region of C3. Our results suggest that hydrolysis may produce a looser (more open) structure in the C3d region, in which some of the changes affect the conversion of helical segments into coil segments facilitating interactions with factors I and H. This study represents the first detailed study mapping the regions of C3 involved in conformational transition when hydrolyzed to C3(H2O). PMID- 15749881 TI - Cryptococcus neoformans capsular glucuronoxylomannan induces expression of fas ligand in macrophages. AB - The major component of capsular material of Cryptococcus neoformans is glucuronoxylomannnan (GXM), a polysaccharide that exhibits potent immunosuppressive properties in vitro and in vivo. The results reported here show that 1) soluble purified GXM induces a prompt, long-lasting, and potent up regulation of Fas ligand (FasL) on macrophages, 2) the up-regulation of FasL is related to induced synthesis and increased mobilization to the cellular surface, 3) this effect is largely mediated by interaction between GXM and TLR4, 4) FasL up-regulation occurs exclusively in GXM-loaded macrophages, 5) macrophages that show up-regulation of FasL induce apoptosis of activated T cells expressing Fas and Jurkat cells that constitutively express Fas, and 6) anti-Fas Abs rescue T cells from apoptosis induced by GXM. Collectively our results reveal novel aspects of the immunoregulatory properties of GXM and suggest that this nontoxic soluble compound could be used to dampen the immune response, to promote or accelerate the death receptor, and to fix FasL expression in a TLR/ligand dependent manner. In the present study, we delineate potential new therapeutic applications for GXM that exploit death receptors as key molecular targets in regulating cell-mediated cytotoxicity, immune homeostasis, and the immunopathology of diseases. PMID- 15749883 TI - Characterization of the chicken C-type lectin-like receptors B-NK and B-lec suggests that the NK complex and the MHC share a common ancestral region. AB - The sequencing of the chicken MHC led to the identification of two open reading frames, designated B-NK and B-lec, that were predicted to encode C-type lectin domains. C-type lectin domains are not encoded in the MHC of any animal described to date; therefore, this observation was completely unexpected, particularly given that the chicken has a "minimal essential MHC." In this study, we describe the initial characterization of the B-NK and B-lec genes, and show that they share greatest homology with C-type lectin-like receptors encoded in the human NK complex (NKC), in particular NKR-P1 and lectin-like transcript 1 (LLT1), respectively. In common with NKR-P1 and LLT1, B-NK and B-lec are located next to each other and transcribed in opposite orientation. Like human NKR-P1, B-NK has a functional inhibitory signaling motif in the cytoplasmic tail and is expressed in NK cells. In contrast, B-lec contains an endocytosis motif in the cytoplasmic tail, and like LLT1, is an early activation Ag. Further analysis leads us to propose that there are four subgroups of C-type lectin-like receptors in the NKC, which arose as a result of duplication events. Moreover, this analysis suggests that the NKC may be considered a fifth paralogous region, and therefore shares an ancient common origin with the MHC. This provides evidence that C-type lectin like receptors were present in the preduplication, primordial MHC region, and suggests that an original function of MHC molecules was for recognition by NK cell receptors encoded nearby. PMID- 15749885 TI - Genes "waiting" for recruitment by the adaptive immune system: the insights from amphioxus. AB - In seeking evidence of the existence of adaptive immune system (AIS) in ancient chordate, cDNA clones of six libraries from a protochordate, the Chinese amphioxus, were sequenced. Although the key molecules such as TCR, MHC, Ig, and RAG in AIS have not been identified from our database, we demonstrated in this study the extensive molecular evidence for the presence of genes homologous to many genes that are involved in AIS directly or indirectly, including some of which may represent the putative precursors of vertebrate AIS-related genes. The comparative analyses of these genes in different model organisms revealed the different fates of these genes during evolution. Their gene expression pattern suggested that the primitive digestive system is the pivotal place of the origin and evolution of the AIS. Our studies support the general statement that AIS appears after the jawless/jawed vertebrate split. However our study further reveals the fact that AIS is in its twilight in amphioxus and the evolution of the molecules in amphioxus are waiting for recruitment by the emergence of AIS. PMID- 15749884 TI - The protooncogene c-Maf is an essential transcription factor for IL-10 gene expression in macrophages. AB - IL-10 is an important immunoregulatory factor. However, our understanding of IL 10 gene regulation remains very limited. In this study, following up on our previous novel finding that the protooncogene c-Maf of the basic leucine zipper family of transcription factors is expressed in monocytes and macrophages, we investigate the role of c-Maf in the transcriptional regulation of IL-10 and the underlying molecular mechanism in macrophages. c-Maf-null macrophages exhibit strongly impaired IL-10 protein production and mRNA expression upon LPS stimulation. Ectopic expression of c-Maf stimulates not only exogenously transfected IL-10 promoter-driven luciferase activity in a dose-dependent manner but also enhances endogenous IL-10 gene expression stimulated by LPS. Both in vitro and in vivo experiments identify a c-Maf response element localized to nucleotides -196/-184 relative to the transcription initiation site in the IL-10 promoter. This site represents an atypical 12-O-tetradecanoate-13-acetate responsive element for musculoaponeurotic fibrosarcoma recognition and functions as an enhancer element in a heterologous and orientation-independent manner. Furthermore, c-Maf is expressed constitutively in resting monocytes/macrophages. IL-4 can up-regulate c-Maf expression, its binding to IL-10 promoter, and dose dependently enhance IL-10 production induced by LPS; moreover, IL-4 failed to enhance LPS-induced IL-10 production in c-Maf-null macrophages. Taken together, these data demonstrate that c-Maf is an indispensable yet constitutive transcription factor for IL-10 gene expression in LPS-activated macrophages, and IL-4 modulates IL-10 production in inflammatory macrophages likely via its ability to induce c-Maf expression. Thus, this study uncovers a novel and important function of c-Maf in macrophages and elucidates its transcriptional mechanism in the regulation of IL-10 gene expression. PMID- 15749887 TI - Lipid rafts associate with intracellular B cell receptors and exhibit a B cell stage-specific protein composition. AB - Lipid rafts serve as platforms for BCR signal transduction. To better define the molecular basis of these membrane microdomains, we used two-dimensional gel electrophoresis and mass spectrometry to characterize lipid raft proteins from mature as well as immature B cell lines. Of 51 specific raft proteins, we identified a total of 18 proteins by peptide mass fingerprinting. Among them, we found vacuolar ATPase subunits alpha-1 and beta-2, vimentin, gamma-actin, mitofilin, and prohibitin. None of these has previously been reported in lipid rafts of B cells. The differential raft association of three proteins, including a novel potential signaling molecule designated swiprosin-1, correlated with the stage-specific sensitivity of B cells to BCR-induced apoptosis. In addition, MHC class II molecules were detected in lipid rafts of mature, but not immature B cells. This intriguing finding points to a role for lipid rafts in regulating Ag presentation during B cell maturation. Finally, a fraction of the BCR in the B cell line CH27 was constitutively present in lipid rafts. Surprisingly, this fraction was neither expressed at the cell surface nor fully O-glycosylated. Thus, we conclude that partitioning the BCR into lipid rafts occurs in the endoplasmic reticulum/cis-Golgi compartment and may represent a control mechanism for surface transport. PMID- 15749886 TI - Molecular basis for the high affinity interaction between the thymic leukemia antigen and the CD8alphaalpha molecule. AB - The mouse thymic leukemia (TL) Ag is a nonclassical MHC class I molecule that binds with higher affinity to CD8alphaalpha than CD8alphabeta. The interaction of CD8alphaalpha with TL is important for lymphocyte regulation in the intestine. Therefore, we studied the molecular basis for TL Ag binding to CD8alphaalpha. The stronger affinity of the TL Ag for CD8alphaalpha is largely mediated by three amino acids on exposed loops of the conserved alpha3 domain. Mutant classical class I molecules substituted with TL Ag amino acids at these positions mimic the ability to interact with CD8alphaalpha and modulate lymphocyte function. These data indicate that small changes in the alpha3 domain of class I molecules potentially can have profound physiologic consequences. PMID- 15749888 TI - A splice variant of the TCR zeta mRNA lacking exon 7 leads to the down-regulation of TCR zeta, the TCR/CD3 complex, and IL-2 production in systemic lupus erythematosus T cells. AB - The reduction or absence of TCR zeta-chain (zeta) expression in patients with systemic lupus erythematosus (SLE) is thought to be a factor in the pathogenesis of SLE. We previously reported a splice variant of zeta mRNA that lacks the 36-bp exon 7 (zeta mRNA/exon 7(-)) and is accompanied by the down-regulation of zeta protein in T cells from SLE patients. In this study, we show that EX7- mutants (MA5.8 cells deficient in zeta protein that have been transfected with zeta mRNA/exon 7(-)) exhibit a reduction in the expression of TCR/CD3 complex and zeta protein on their cell surface as well as a reduction in the production of IL-2 after stimulation with anti-CD3 Ab, compared with that in wild-type (WT) mutants (MA5.8 cells transfected with the WT zeta mRNA). Furthermore, real-time PCR analyses demonstrated that zeta mRNA/exon 7(-) in EX7- mutants was easily degraded compared with zeta mRNA by the WT mutants. Pulse-chase experiment showed zeta protein produced by this EX7- mutants was more rapidly decreased compared with the WT mutants. Thus, the lower stability of zeta mRNA/exon 7(-) might also be responsible for the reduced expression of the TCR/CD3 complex, including zeta protein, in SLE T cells. PMID- 15749889 TI - Functional domains of Runx1 are differentially required for CD4 repression, TCRbeta expression, and CD4/8 double-negative to CD4/8 double-positive transition in thymocyte development. AB - Runx1 (AML1) has multiple functions in thymocyte development, including CD4 repression in immature thymocytes, expression of TCRbeta, and efficient beta selection. To determine the functional domains of Runx1 important for thymocyte development, we cultured Runx1-deficient murine fetal liver (FL) cells on OP9 Delta-like 1 murine stromal cells, which express Delta-like 1 and support thymocyte development in vitro, and introduced Runx1 or C-terminal-deletion mutants of Runx1 into the FL cells by retrovirus infection. In this system, Runx1 deficient FL cells failed to follow normal thymocyte development, whereas the introduction of Runx1 into the cells was sufficient to produce thymocyte development that was indistinguishable from that in wild-type FL cells. In contrast, Runx1 mutants that lacked the activation domain necessary for initiating gene transcription did not fully restore thymocyte differentiation, in that it neither repressed CD4 expression nor promoted the CD4/8 double-negative to CD4/8 double-positive transition. Although the C-terminal VWRPY motif deficient mutant of Runx1, which cannot interact with the transcriptional corepressor Transducin-like enhancer of split (TLE), promoted the double-negative to double-positive transition, it did not efficiently repress CD4 expression. These results suggest that the activation domain is essential for Runx1 to establish thymocyte development and that Runx1 has both TLE-dependent and TLE independent functions in thymocyte development. PMID- 15749890 TI - The IL-27 receptor chain WSX-1 differentially regulates antibacterial immunity and survival during experimental tuberculosis. AB - IL-12 is a potent inducer of IFN-gamma production and promotes a protective cell mediated immune response after Mycobacterium tuberculosis infection. Recently, the IL-12-related cytokine IL-27 was discovered, and WSX-1 was identified as one component of the IL-27R complex. To determine the functional significance of IL 27/WSX-1 during tuberculosis, we analyzed the course of infection and the immune response in WSX-1-KO mice after aerosol infection with M. tuberculosis. In the absence of WSX-1, an increased production of the proinflammatory cytokines TNF and IL-12p40 resulted in elevated CD4+ T cell activation and IFN-gamma production, which enhanced macrophage effector functions and reduced bacterial loads. This is the first occasion of a selectively gene-deficient mouse strain showing higher levels of protective immunity against M. tuberculosis infection than wild-type mice. However, a concomitantly increased chronic inflammatory response also accelerated death of infected WSX-1-KO mice. In vitro, IL-27 induced STAT3 phosphorylation and inhibited TNF and IL-12 production in activated peritoneal macrophages, indicating a novel feedback mechanism by which IL-27 can modulate excessive inflammation. In conclusion, IL-27 both prevents optimal antimycobacterial protection and limits the pathological sequelae of chronic inflammation. PMID- 15749891 TI - Neisserial porin-induced dendritic cell activation is MyD88 and TLR2 dependent. AB - Neisserial porins have been shown to act as B cell mitogens and immune adjuvants. PorA and PorB are the major outer membrane porin proteins of the human pathogen Neisseria meningitidis. We have shown that the mechanism of the immunopotentiating capability of porin involves up-regulation of the T cell costimulatory ligand, CD86. Due to neisserial porin's ability to activate B cells and potentiate immune responses, we hypothesized that porin also employs the potent immune stimulatory function of dendritic cells (DC). We examined the ability of purified N. meningitidis PorB to induce maturation of murine splenic and bone marrow-derived DC. PorB treatment induced DC maturation, as demonstrated by increased expression of CD86 and class I and II MHC molecules. In addition, PorB not only enhanced the allostimulatory activity of DC, but also augmented the ability of DC to stimulate T cells in an Ag-specific manner. PorB-matured DC secreted the inflammatory cytokine IL-6, which may have implications for the adjuvant property of porin. Induction of IL-6 by PorB is also significant because IL-6 is one of a number of cytokines produced during infection with N. meningitidis and may be involved in the inflammatory process observed during infection and disease. We previously demonstrated the requirement of MyD88 and TLR2 for PorB-induced B cell activation. In the present study, MyD88 and TLR2 were also essential for PorB-induced DC activation. This work is significant for elucidating the mechanism(s) of neisserial porin's immune stimulatory activity. PMID- 15749892 TI - Sphingosine kinase 1 (SK1) is recruited to nascent phagosomes in human macrophages: inhibition of SK1 translocation by Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis (M.tb) is a leading cause of global infectious mortality. The pathogenesis of tuberculosis involves inhibition of phagosome maturation, leading to survival of M.tb within human macrophages. A key determinant is M.tb-induced inhibition of macrophage sphingosine kinase (SK) activity, which normally induces Ca2+ signaling and phagosome maturation. Our objective was to determine the spatial localization of SK during phagocytosis and its inhibition by M.tb. Stimulation of SK activity by killed M.tb, live Staphylococcus aureus, or latex beads was associated with translocation of cytosolic SK1 to the phagosome membrane. In contrast, SK1 did not associate with phagosomes containing live M.tb. To characterize the mechanism of phagosomal translocation, live cell confocal microscopy was used to compare the localization of wild-type SK1, catalytically inactive SK1G82D, and a phosphorylation-defective mutant that does not undergo plasma membrane translocation (SK1S225A). The magnitude and kinetics of translocation of SK1G82D and SK1S225A to latex bead phagosomes were indistinguishable from those of wild-type SK1, indicating that novel determinants regulate the association of SK1 with nascent phagosomes. These data are consistent with a model in which M.tb inhibits both the activation and phagosomal translocation of SK1 to block the localized Ca2+ transients required for phagosome maturation. PMID- 15749893 TI - Burkholderia cenocepacia induces neutrophil necrosis in chronic granulomatous disease. AB - Burkholderia cepacia complex is a life-threatening group of pathogens for patients with chronic granulomatous disease (CGD), whose phagocytes are unable to produce reactive oxygen species (ROS). Unlike other CGD pathogens, B. cepacia complex is particularly virulent, characteristically causing septicemia, and is the bacterial species responsible for most fatalities in these patients. We found that a nonmucoid Burkholderia cenocepacia (a predominant species in the B. cepacia complex) isolate was readily ingested by normal human neutrophils under nonopsonic conditions and promoted apoptosis in these cells. The proapoptotic effect was not due to secreted bacterial products, but was dependent on bacterial viability. Phagocytosis was associated with a robust production of ROS, and the apoptotic neutrophils could be effectively cleared by monocyte-derived macrophages. The proapoptotic effect of B. cenocepacia was independent of ROS production because neutrophils from CGD patients were rendered apoptotic to a similar degree as control cells after challenge. More importantly, neutrophils from CGD patients, but not from normal individuals, were rendered necrotic after phagocytosis of B. cenocepacia. The extreme virulence of B. cepacia complex bacteria in CGD, but not in immunocompetent hosts, could be due to its necrotic potential in the absence of ROS. PMID- 15749894 TI - Influence of ESAT-6 secretion system 1 (RD1) of Mycobacterium tuberculosis on the interaction between mycobacteria and the host immune system. AB - The chromosomal locus encoding the early secreted antigenic target, 6 kDa (ESAT 6) secretion system 1 of Mycobacterium tuberculosis, also referred to as "region of difference 1 (RD1)," is absent from Mycobacterium bovis bacillus Calmette Guerin (BCG). In this study, using low-dose aerosol infection in mice, we demonstrate that BCG complemented with RD1 (BCG::RD1) displays markedly increased virulence which albeit does not attain that of M. tuberculosis H37Rv. Nevertheless, phenotypic and functional analyses of immune cells at the site of infection show that the capacity of BCG::RD1 to initiate recruitment/activation of immune cells is comparable to that of fully virulent H37Rv. Indeed, in contrast to the parental BCG, BCG::RD1 mimics H37Rv and induces substantial influx of activated (CD44highCD45RB(-)CD62L(-)) or effector (CD45RB(-)CD27(-)) T cells and of activated CD11c(+)CD11bhigh cells to the lungs of aerosol-infected mice. For the first time, using in vivo analysis of transcriptome of inflammatory cytokines and chemokines of lung interstitial CD11c+ cells, we show that in a low dose aerosol infection model, BCG::RD1 triggered an activation/inflammation program comparable to that induced by H37Rv while parental BCG, due to its overattenuation, did not initiate the activation program in lung interstitial CD11c+ cells. Thus, products encoded by the ESAT-6 secretion system 1 of M. tuberculosis profoundly modify the interaction between mycobacteria and the host innate and adaptive immune system. These modifications can explain the previously described improved protective capacity of BCG::RD1 vaccine candidate against M. tuberculosis challenge. PMID- 15749895 TI - Reovirus serotype 1/strain Lang-stimulated activation of antigen-specific T lymphocytes in Peyer's patches and distal gut-mucosal sites: activation status and cytotoxic mechanisms. AB - Intraduodenal priming of mice with reovirus serotype 1/strain Lang (reovirus 1/L) stimulates gut lymphocytes and generates precursor and effector CTLs. Our earlier studies demonstrated that germinal center and T cell Ag (GCT) is a marker which identifies reovirus 1/L-specific precursor CTL and effector CTL in Peyer's patches (PP) of reovirus 1/L-inoculated mice. In this study, we characterized the expression of the activation markers, GCT and CD11c, on reovirus 1/L-stimulated gut lymphocytes and the effector mechanisms involved in reovirus 1/L-specific cytotoxicity. We found that intraduodenal reovirus 1/L inoculation of mice induced the expression of both GCT and CD11c on PP lymphocytes (PPL), intraepithelial lymphocytes (IEL), and lamina propria lymphocytes (LPL), and these activated cells expressed Fas ligand (FasL). The majority of the GCT+ CD11c+ IEL and LPL expressed a phenotype, TCRalphabeta+ Thy-1+ CD8+ similar to that expressed on reovirus 1/L-stimulated PPL. However, splenic lymphocytes expressed GCT but not CD11c after stimulation with reovirus 1/L. Perforin, Fas FasL, and TRAIL pathways were found to be involved in PPL, IEL, and LPL cytotoxic activity against reovirus 1/L-infected targets. In PPL, perforin and Fas-FasL pathways were more effective than TRAIL. In IEL, all three cytotoxic mechanisms were equally as effective. However, LPL prefer Fas-FasL and TRAIL over perforin. Further, we demonstrated the preferential migration of GCT+ PPL to the intraepithelial compartment and the lamina propria. These results suggest that GCT and CD11c can be used as activation markers for gut lymphocytes and CD11c can also be used to differentiate between activated gut and systemic lymphocytes. PMID- 15749896 TI - A novel role of IL-15 in early activation of memory CD8+ CTL after reinfection. AB - A rapid induction of effector functions in memory T cells provides rapid and intensified protection against reinfection. To determine potential roles of IL-15 in early expansion and activation of memory CD8+ T cells in secondary immune response, we examined the cell division and cytotoxicity of memory CD8+ T cells expressing OVA(257-264)/Kb-specific TCR that were transferred into IL-15 transgenic (Tg) mice, IL-15 knockout (KO) mice, or control C57BL/6 mice followed by challenge with recombinant Listeria monocytogenes expressing OVA (rLM-OVA). In vivo CTL activities and expression of granzyme B of the transferred CD8+ T cells were significantly higher in the IL-15 Tg mice but lower in the IL-15 KO mice than those in control mice at the early stage after challenge with rLM-OVA. In contrast, there was no difference in the cell division in IL-15 Tg mice and IL-15 KO mice compared with those in control mice. In vivo administration of rIL-15 conferred robust protection against reinfection via induction of granzyme B in the memory CD8+ T cells. These results suggest that IL-15 plays an important role in early activation of memory CD8+ T cells. PMID- 15749897 TI - Mechanisms of the natural reactivity of lymphocytes from noninfected individuals to membrane-associated Leishmania infantum antigens. AB - Membrane-associated Leishmania Ags (MLA) or soluble Leishmania Ags were used in vitro to stimulate cord blood or PBMC from healthy donors noninfected by Leishmania parasites. MLA, but not soluble Leishmania Ags, constantly induce strong proliferation of cord blood mononuclear cells and PBMC from noninfected individuals. Responding cells are CD3+, CD4+, TCRalphabeta+, CD45RO+, and CD45RA+ and secrete IFN-gamma and IL-10, but not IL-4. MLA do not activate NK cells nor NKT cells. Membrane Ags also induce purified macrophages from noninfected individuals to secrete IL-10 and TNF-alpha, but have no effect on IL-1alpha or IL 12 secretion. The effects of MLA are proteinase K-sensitive and resistant to lipid extraction. The lymphoproliferative responses are inhibited by anti-HLA-DR Abs and require Ag processing by APCs, excluding that the biological effect of MLA could be attributed to a superantigen. Finally, TCR repertoire analysis shows that the T cell expansion induced by MLA uses TCR with various variable beta segment rearrangements and CDR3 lengths, features much more characteristic to those observed with a polyclonal activator than with a conventional Ag. These results suggest a particular mechanism developed during the host's natural response to Leishmania parasites that allows direct activation of naive CD4 lymphocytes by parasite membrane-associated Ags. PMID- 15749898 TI - Intracellular IL-1 receptor antagonist type 1 inhibits IL-1-induced cytokine production in keratinocytes through binding to the third component of the COP9 signalosome. AB - The IL-1 receptor antagonist (IL-1Ra) exists in four isoforms, three of which lack signal peptides and are primarily intracellular proteins. The biologic roles of the intracellular isoforms of IL-1Ra have remained unknown. The objective of these studies was to determine whether the major intracellular isoform of IL-1Ra 18-kDa type 1 (icIL-1Ra1), mediated unique functions inside cells. A yeast two hybrid screen with HeLa cell lysates revealed specific binding of icIL-1Ra1, and not of the other IL-1Ra isoforms, to the third component of the COP9 signalosome complex (CSN3). This binding was confirmed by Far Western blot analysis, sedimentation on a glycerol gradient, glutathione pull-down experiments, and coimmunoprecipitation. In addition to binding specifically to CSN3, icIL-1Ra1 inhibited phosphorylation of p53, c-Jun, and IkappaB by the crude CSN-associated kinase and of p53 by recombinant protein kinase CK2 and protein kinase D, both associated with CSN3. The biologic relevance of the interaction between icIL-1Ra1 and CSN3 was demonstrated in the keratinocyte cell lines KB and A431, both possessing abundant CSN3. A431 cells exhibited high levels of icIL-1Ra1 but lacked both detectable IL-1alpha-induced IL-6 and IL-8 production and phosphorylation of p38 MAPK. KB cells displayed the opposite pattern which was reversed after transfection with icIL-1Ra1 mRNA. Inhibition of CSN3 or of icIL 1Ra1 production through gene knockdown with specific small interfering RNA in A431 cells each led to an inhibition of IL-1alpha-induced IL-6 and IL-8 production. Thus, icIL-1Ra1 exhibits unique anti-inflammatory properties inside cells through binding to CSN3 with subsequent inhibition of the p38 MAPK signal transduction pathway. PMID- 15749899 TI - Involvement of BLT1 endocytosis and Yes kinase activation in leukotriene B4 induced neutrophil degranulation. AB - One of the important biological activities of human neutrophils is degranulation, which can be induced by leukotriene B4 (LTB4). Here we investigated the intracellular signaling events involved in neutrophil degranulation mediated by the high affinity LTB4 receptor, BLT1. Peripheral blood neutrophils as well as the promyeloid PLB-985 cell line, stably transfected with BLT1 cDNA and differentiated into a neutrophil-like cell phenotype, were used throughout this study. LTB4-induced enzyme release was inhibited by 50-80% when cells were pretreated with the pharmacological inhibitors of endocytosis sucrose, Con A and NH4Cl. In addition, transient transfection with a dominant negative form of dynamin (K44A) resulted in approximately 70% inhibition of ligand-induced degranulation. Pretreating neutrophils or BLT1-expressing PLB-985 cells with the Src family kinase inhibitor PP1 resulted in a 30-60% inhibition in BLT1-mediated degranulation. Yes kinase, but not c-Src, Fgr, Hck, or Lyn, was found to exhibit up-regulated kinase activity after LTB4 stimulation. Moreover, BLT1 endocytosis was found to be necessary for Yes kinase activation in neutrophils. LTB4-induced degranulation was also sensitive to inhibition of PI3K. In contrast, it was not affected by inhibition of the mitogen-activated protein kinase MEK kinase, the Janus kinases, or the receptor tyrosine kinase epidermal growth factor receptor or platelet-derived growth factor receptor. Taken together, our results suggest an essential role for BLT1 endocytosis and Yes kinase activation in LTB4-mediated degranulation of human neutrophils. PMID- 15749900 TI - Stem cell factor has a suppressive activity to IgE-mediated chemotaxis of mast cells. AB - Stem cell factor (SCF), which is well known as a cytokine capable of amplifying development and functions of mast cells, is mainly released from fibroblasts in the peripheral tissue. To investigate whether SCF controlled chemotactic migration of mast cells induced by IgE-specific Ag, murine bone marrow-derived cultured mast cells (BMCMC) and human cord blood-derived cultured mast cells (HuCMC) were preincubated with SCF. Although BMCMC and HuCMC sensitized with IgE directly moved toward specific Ag, preincubation for even 1 h with an optimal dose of SCF suppressed the IgE-mediated chemotactic movement. No or little inhibitory effect of SCF was detected in BMCMC derived from c-kit receptor-defect WBB6F1-W/Wv mice. In contrast, preincubation of BMCMC and HuCMC with SCF enhanced beta-hexosaminidase release and Ca2+ mobilization in response to Ag after sensitization with IgE. Using the real-time record of chemotactic migration, BMCMC preincubated with SCF manifested motionless without degranulation. These results suggest that locally produced SCF may have an inhibitory effect on chemotaxis of mast cells, contributing to their accumulation and enhancement of functions at the peripheral site in allergic and nonallergic conditions. PMID- 15749901 TI - Inhibition of neutrophil apoptosis by TLR agonists in whole blood: involvement of the phosphoinositide 3-kinase/Akt and NF-kappaB signaling pathways, leading to increased levels of Mcl-1, A1, and phosphorylated Bad. AB - Using flow cytometry, we investigated the effect of TLR agonists on human polymorphonuclear neutrophil (PMN) apoptosis in whole blood. LPS (TLR4), peptidoglycan (TLR2), R-848 (TLR7/8), and CpG-DNA (TLR9) were equally effective at delaying spontaneous apoptosis of PMN, while PamCSK4 (TLR1/2), macrophage activating lipopeptide-2 (TLR2/6), flagellin (TLR5), and loxoribine (TLR7) were less effective or inactive. TLR agonists found to delay apoptosis also extended the functional life span of PMN. Analysis of signaling pathways revealed that the antiapoptotic effect of TLR agonists required NF-kappaB and PI3K activation. Furthermore, analysis of intact cells by flow cytometry showed that TLR agonists delaying PMN apoptosis increased phosphorylation of Akt, a major target of PI3K. This effect was associated with a PI3K-dependent increase in heat shock protein 27 phosphorylation, which has been reported to play a key role in PMN survival. Finally, the TLR-induced delay in PMN apoptosis was associated with increased levels of Mcl-1 and A1, which are antiapoptotic members of the Bcl-2 family. These effects were reversed by PI3K and NF-kappaB inhibitors, respectively. TLR activation also led to PI3K-dependent phosphorylation of the proapoptotic protein Bad. Taken together, our results strongly suggest a role of NF-kappaB and PI3K in TLR-induced PMN survival, leading to modulation of Bcl-2 family molecules. PMID- 15749902 TI - Pyocyanin production by Pseudomonas aeruginosa induces neutrophil apoptosis and impairs neutrophil-mediated host defenses in vivo. AB - Clearance of neutrophils from inflamed sites is critical for resolution of inflammation, but pathogen-driven neutrophil apoptosis can impair host defenses. We previously showed that pyocyanin, a phenazine toxic metabolite produced by Pseudomonas aeruginosa, accelerates neutrophil apoptosis in vitro. We compared wild-type and pyocyanin-deficient strains of P. aeruginosa in a murine model of acute pneumonia. Intratracheal instillation of either strain of P. aeruginosa caused a rapid increase in bronchoalveolar lavage neutrophil counts up to 18 h after infection. In wild-type infection, neutrophil numbers then declined steadily, whereas neutrophil numbers increased up to 48 h in mice infected with pyocyanin-deficient P. aeruginosa. In keeping with these differences, pyocyanin production was associated with reduced bacterial clearance from the lungs. Neutrophil apoptosis was increased in mice infected with wild-type compared with the phenazine-deficient strain or two further strains that lack pyocyanin production, but produce other phenazines. Concentrations of potent neutrophil chemokines (MIP-2, KC) and cytokines (IL-6, IL-1beta) were significantly lower in wild-type compared with phenazine-deficient strain-infected mice at 18 h. We conclude that pyocyanin production by P. aeruginosa suppresses the acute inflammatory response by pathogen-driven acceleration of neutrophil apoptosis and by reducing local inflammation, and that this is advantageous for bacterial survival. PMID- 15749903 TI - The nuclear IkappaB protein IkappaBNS selectively inhibits lipopolysaccharide induced IL-6 production in macrophages of the colonic lamina propria. AB - Macrophages play an important role in the pathogenesis of chronic colitis. However, it remains unknown how macrophages residing in the colonic lamina propria are regulated. We characterized colonic lamina proprial CD11b-positive cells (CLPMphi). CLPMphi of wild-type mice, but not IL-10-deficient mice, displayed hyporesponsiveness to TLR stimulation in terms of cytokine production and costimulatory molecule expression. We compared CLPMphi gene expression profiles of wild-type mice with IL-10-deficient mice, and identified genes that are selectively expressed in wild-type CLPMphi. These genes included nuclear IkappaB proteins such as Bcl-3 and IkappaBNS. Because Bcl-3 has been shown to specifically inhibit LPS-induced TNF-alpha production, we analyzed the role of IkappaBNS in macrophages. Lentiviral introduction of IkappaBNS resulted in impaired LPS-induced IL-6 production, but not TNF-alpha production in the murine macrophage cell line RAW264.7. IkappaBNS expression led to constitutive and intense DNA binding of NF-kappaB p50/p50 homodimers. IkappaBNS was recruited to the IL-6 promoter, but not to the TNF-alpha promoter, together with p50. Furthermore, small interference RNA-mediated reduction in IkappaBNS expression in RAW264.7 cells resulted in increased LPS-induced production of IL-6, but not TNF alpha. Thus, IkappaBNS selectively suppresses LPS-induced IL-6 production in macrophages. This study established that nuclear IkappaB proteins differentially regulate LPS-induced inflammatory cytokine production in macrophages. PMID- 15749904 TI - Helicobacter pylori disrupts NADPH oxidase targeting in human neutrophils to induce extracellular superoxide release. AB - Helicobacter pylori (Hp) infection triggers a chronic influx of polymorphonuclear leukocyte neutrophils (PMNs) into the gastric mucosa. Although Hp reside in a neutrophil-rich environment, how these organisms evade phagocytic killing is largely unexplored. We now show that live Hp (strains 11637, 60190, DT61A, and 11916) are readily ingested by PMNs and induce a rapid and strong respiratory burst that is comparable to PMA. Relative to other particulate stimuli, Hp are more potent activators of PMNs than opsonized zymosan, Staphylococcus aureus, or Salmonella. Strikingly, biochemical and microscopic analyses demonstrate that Hp disrupt NADPH oxidase targeting such that superoxide anions are released into the extracellular milieu and do not accumulate inside Hp phagosomes. Specifically, nascent Hp phagosomes acquire flavocytochrome b558 but do not efficiently recruit or retain p47phox or p67phox. Superoxide release peaks at 16 min coincident with the appearance of assembled oxidase complexes in patches at the cell surface. Oxidant release is regulated by formalin-resistant and heat-sensitive bacterial surface factors distinct from urease and Hp(2-20). Following opsonization with fresh serum, Hp triggers a modest respiratory burst that is confined to the phagosome, and ingested bacteria are eliminated. We conclude that disruption of NADPH oxidase targeting allows unopsonized Hp to escape phagocytic killing, and our findings support the hypothesis that bacteria and PMNs act in concert to damage the gastric mucosa. PMID- 15749905 TI - Manifestations of inflammatory arthritis are critically dependent on LFA-1. AB - Leukocyte infiltration of synovial fluid and tissues is the hallmark of inflammatory arthritis. Selectins and beta2 integrins have been implicated in the multistep process of leukocyte adhesion to vascular endothelium. However, previous work has revealed disparate requirements for leukocyte recruitments to specific anatomic locales. Moreover, the mechanisms regulating recruitment of leukocytes to the joint in inflammatory arthritis models are not fully understood. We hypothesized that beta2 integrins, expressed on leukocytes, might play a pathogenic role in synovial inflammation. Using mice deficient in all beta2 integrins (CD18 null mice), we demonstrate that expression of these heterodimeric adhesion molecules is critical for arthritis induction in the K/B x N serum transfer model. Using null-allele mice and blocking mAbs, we demonstrate specifically that CD11a/CD18 (LFA-1) is absolutely required for the development of arthritis in this model. Blocking mAbs further revealed an ongoing requirement for LFA-1 I-domain adhesive function in disease perpetuation. These findings suggest that the LFA-1 I-domain forms an attractive target for treatment of human inflammatory arthritis. PMID- 15749906 TI - Febrile-range hyperthermia augments neutrophil accumulation and enhances lung injury in experimental gram-negative bacterial pneumonia. AB - We previously demonstrated that exposure to febrile-range hyperthermia (FRH) accelerates pathogen clearance and increases survival in murine experimental Klebsiella pneumoniae peritonitis. However, FRH accelerates lethal lung injury in a mouse model of pulmonary oxygen toxicity, suggesting that the lung may be particularly susceptible to injurious effects of FRH. In the present study, we tested the hypothesis that, in contrast with the salutary effect of FRH in Gram negative peritonitis, FRH would be detrimental in multilobar Gram-negative pneumonia. Using a conscious, temperature-clamped mouse model and intratracheal inoculation with K. pneumoniae Caroli strain, we showed that FRH tended to reduce survival despite reducing the 3 day-postinoculation pulmonary pathogen burden by 400-fold. We showed that antibiotic treatment rescued the euthermic mice, but did not reduce lethality in the FRH mice. Using an intratracheal bacterial endotoxin LPS challenge model, we found that the reduced survival in FRH-treated mice was accompanied by increased pulmonary vascular endothelial injury, enhanced pulmonary accumulation of neutrophils, increased levels of IL-1beta, MIP 2/CXCL213, GM-CSF, and KC/CXCL1 in the bronchoalveolar lavage fluid, and bronchiolar epithelial necrosis. These results suggest that FRH enhances innate host defense against infection, in part, by augmenting polymorphonuclear cell delivery to the site of infection. The ultimate effect of FRH is determined by the balance between accelerated pathogen clearance and collateral tissue injury, which is determined, in part, by the site of infection. PMID- 15749907 TI - The human herpes virus 8-encoded chemokine receptor is required for angioproliferation in a murine model of Kaposi's sarcoma. AB - Kaposi's sarcoma (KS)-associated herpesvirus or human herpes virus 8 is considered the etiological agent of KS, a highly vascularized neoplasm that is the most common tumor affecting HIV/AIDS patients. The KS-associated herpesvirus/human herpes virus 8 open reading frame 74 encodes a constitutively active G protein-coupled receptor known as vGPCR that binds CXC chemokines with high affinity. In this study, we show that conditional transgenic expression of vGPCR by cells of endothelial origin triggers an angiogenic program in vivo, leading to development of an angioproliferative disease that resembles KS. This angiogenic program consists partly in the expression of the angiogenic factors placental growth factor, platelet-derived growth factor B, and inducible NO synthase by the vGPCR-expressing cells. Finally, we show that continued vGPCR expression is essential for progression of the KS-like phenotype and that down regulation of vGPCR expression results in reduced expression of angiogenic factors and regression of the lesions. Together, these findings implicate vGPCR as a key element in KS pathogenesis and suggest that strategies to block its function may represent a novel approach for the treatment of KS. PMID- 15749908 TI - IL-22 inhibits epidermal differentiation and induces proinflammatory gene expression and migration of human keratinocytes. AB - IL-22 belongs to a family of cytokines structurally related to IL-10, including IL-19, IL-20, IL-24, and IL-26. In contrast to IL-10, IL-22 has proinflammatory activities. IL-22 signals through a class II cytokine receptor composed of an IL 22-binding chain, IL-22RA1, and the IL-10RB subunit, which is shared with the IL 10R. In the present study, we show that short-term cultured human epidermal keratinocytes express a functional IL-22R but no IL-10R. Accordingly, IL-22 but not IL-10 induces STAT3 activation in keratinocytes. Using a cDNA array screening approach, real-time RT-PCR, and Western blot analysis, we demonstrate that IL-22 up-regulates, in a dose-dependent manner, the expression of S100A7, S100A8, S100A9, a group of proinflammatory molecules belonging to the S100 family of calcium-binding proteins, as well as the matrix metalloproteinase 3, the platelet derived growth factor A, and the CXCL5 chemokine. In addition, IL-22 induces keratinocyte migration in an in vitro injury model and down-regulates the expression of at least seven genes associated with keratinocyte differentiation. Finally, we show that IL-22 strongly induces hyperplasia of reconstituted human epidermis. Taken together, these results suggest that IL-22 plays an important role in skin inflammatory processes and wound healing. PMID- 15749909 TI - Activation of the prostaglandin D2 receptor DP2/CRTH2 increases allergic inflammation in mouse. AB - Allergic pathologies are often associated with IgE production, mast cell activation, and eosinophilia. PGD2 is the major eicosanoid, among several inflammatory mediators, released by mast cells. PGD2 binds to two membrane receptors, D prostanoid receptor (DP)1 and DP2, endowed with antagonistic properties. In humans, DP2 is preferentially expressed on type 2 lymphocytes, eosinophils, and basophils and mediates chemotaxis in vitro. Although not yet supported by in vivo studies, DP2 is thought to be important in the promotion of Th2-related inflammation. Herein, we demonstrate that mouse eosinophils express both DP1 and DP2 and that PGD2 exerts in vitro chemotactic effects on eosinophils through DP2 activation. Furthermore, 13,14-dihydro-15-keto-PGD2, a specific DP2 agonist not only increases eosinophil recruitment at inflammatory sites but also the pathology in two in vivo models of allergic inflammation: atopic dermatitis and allergic asthma. By contrast, DP1 activation tends to ameliorate the pathology in asthma. Taken together, these results support the hypothesis that DP2 might play a critical role in allergic diseases and underline the interest of DP2 antagonists in human therapy. PMID- 15749910 TI - Unconjugated bilirubin inhibits VCAM-1-mediated transendothelial leukocyte migration. AB - During lymphocyte migration, engagement of VCAM-1 stimulates the generation of endothelial cell-derived reactive oxygen species (ROS) and activation of matrix metalloproteinases, facilitating endothelial retraction. Because bilirubin is a potent antioxidant, we examined the hypothesis that this bile pigment inhibits VCAM-1-dependent cellular events. The migration of isolated murine splenic lymphocytes across monolayers of murine endothelial cell lines (which constitutively express VCAM-1) is significantly inhibited by physiological concentrations of bilirubin, in the absence of an effect on lymphocyte adhesion. Bilirubin administration also suppresses VCAM-1-stimulated ROS generation and reduces endothelial cell matrix metalloproteinase activity. In a murine asthma model characterized by VCAM-1-dependent airway inflammation, treatment of C57BL6/J mice with i.p. bilirubin decreases the total leukocyte count in the lung parenchyma and lavage fluid, through specific inhibition of eosinophil and lymphocyte infiltration. Blood eosinophil counts were increased in bilirubin treated animals, while VCAM-1 expression in the capillary endothelium and cytokine levels in both lung lavage and supernatants from cultured lymph node lymphocytes were unchanged, suggesting that bilirubin inhibits leukocyte migration. CONCLUSION: bilirubin blocks VCAM-1-dependent lymphocyte migration in vitro and ameliorates VCAM-1-mediated airway inflammation in vivo, apparently through the suppression of cellular ROS production. These findings support a potential role for bilirubin as an endogenous immunomodulatory agent. PMID- 15749911 TI - The cytokine IL-1beta activates IFN response factor 3 in human fetal astrocytes in culture. AB - The cytokine IL-1beta is a major activator of primary human fetal astrocytes in culture, leading to the production of a wide range of cytokines and chemokines important in the host defense against pathogens. IL-1beta, like TLR4, signals via the MyD88/IL-1betaR-associated kinase-1 pathway linked to activation of NF-kappaB and AP-1. Recent studies have shown that TLR4 also signals independently of MyD88, resulting in the activation of IFN regulatory factor 3 (IRF3), a transcription factor required for the production of primary antiviral response genes such as IFN-beta. Using a functional genomics approach, we observed that IL 1beta induced in astrocytes a group of genes considered to be IFN-stimulated genes (ISG), suggesting that IL-1beta may also signal via IRF3 in these cells. We now show, using real-time PCR, that in astrocytes IL-1beta induces the expression of IFN-beta, IRF7, CXCL10/IFN-gamma-inducible protein-10, and CCL5/RANTES. Chemokine expression was confirmed by ELISA. We also show that IL-1beta induces phosphorylation and nuclear translocation of IRF3 and delayed phosphorylation of STAT1. The dependency of IFN-beta, IRF7, and CXCL10/IFN-gamma-inducible protein 10 gene expression on IRF3 was confirmed using a dominant negative IRF3 expressing adenovirus. The robust induction by IL-1beta of additional ISG noted on the microarrays, such as STAT1, 2'5'-oligoadenylate synthetase 2, and ISG15, also supports an active signaling role for IL-1beta via this pathway in human fetal astrocytes. These data are the first to show that IL-1beta, in addition to TLRs, can stimulate IRF3, implicating this cytokine as an activator of genes involved in innate antiviral responses in astrocytes. PMID- 15749912 TI - Modulation of phagocytosis of apoptotic neutrophils by supernatant from dexamethasone-treated macrophages and annexin-derived peptide Ac(2-26). AB - Phagocytic clearance of apoptotic leukocytes plays an important role in the resolution of inflammation. The glucocorticoid-inducible protein annexin 1 and annexin 1-derived peptides show potent anti-inflammatory responses in acute and chronic inflammation. In this study, we report that the annexin 1-derived peptide (Ac(2-26)) significantly stimulates nonphlogistic phagocytosis of apoptotic polymorphonuclear leukocytes (PMNs) by human monocyte-derived macrophages (Mphi). Peptide Ac(2-26)-stimulated phagocytosis is accompanied by rearrangement of the Mphi actin cytoskeleton. To investigate the potential role of endogenous annexin on clearance of apoptotic cells, Mphi were cultured for 5 days in the presence of dexamethasone. Supernatants collected from dexamethasone-treated Mphi significantly enhanced the ability of naive Mphi to engulf apoptotic PMNs. This effect was blocked by an annexin blocking Ab, by immunodepletion of the supernatants, and by the formyl peptide receptor/lipoxin receptor antagonist Boc1. In addition, we show that bone marrow-derived Mphi from annexin 1-null mice present a 40% decreased phagocytosis of apoptotic PMNs compared with cells taken from littermate controls. In conclusion, these results emphasize the pivotal role of annexin 1 as mediator for clearance of apoptotic cells and expand its potential therapeutic role in controlling inflammatory diseases. PMID- 15749913 TI - Indispensable role of Stat5a in Stat6-independent Th2 cell differentiation and allergic airway inflammation. AB - It is well-recognized that Stat6 plays a critical role in Th2 cell differentiation and the induction of allergic inflammation. We have previously shown that Stat5a is also required for Th2 cell differentiation and allergic airway inflammation. However, it is the relative importance and redundancy of Stat6 and Stat5a in Th2 cell differentiation and allergic airway inflammation are unknown. In this study we addressed these issues by comparing Stat5a-deficient (Stat5a(-/-)) mice, Stat6(-/-) mice, and Stat5a- and Stat6 double-deficient (Stat5a(-/-) Stat6(-/-)) mice on the same genetic background. Th2 cell differentiation was severely decreased in Stat6(-/-)CD4+ T cells, but Stat6 independent Th2 cell differentiation was still significantly observed in Stat6(-/ )CD4+ T cells. However, even in the Th2-polarizing condition (IL-4 plus anti-IFN gamma mAb), no Th2 cells developed in Stat5a(-/-)Stat6(-/-) CD4+ T cells. Moreover, Ag-induced eosinophil and lymphocyte recruitment in the airways was severely decreased in Stat5a(-/-)Stat6(-/-) mice compared with that in Stat6(-/-) mice. These results indicate that Stat5a plays an indispensable role in Stat6 independent Th2 cell differentiation and subsequent Th2 cell-mediated allergic airway inflammation. PMID- 15749915 TI - B cell response to surface IgM cross-linking identifies different prognostic groups of B-chronic lymphocytic leukemia patients. AB - On the basis of responses to surface IgM (sIgM) cross-linking, B cells from 41 patients with B-chronic lymphocytic leukemia were categorized as 15 nonresponders (group I) and 26 responders (group II). The latter cases were subclassified as those seven where proliferation was induced (subgroup IIa) and the remaining 19 in whom apoptosis occurred (subgroup IIa). Signal disruption in group I was confirmed by the absence of Ca2+ mobilization. Activation of PI3K was constitutive in subgroup IIa, but not in subgroup IIb, and that of Akt induced by anti-mu in subgroup IIa, but not in subgroup IIb. Among the MAPK, ERK was more highly activated relative to p38 in subgroup IIa, whereas activation of p38 predominated over that of ERK in subgroup IIb. For subgroup IIb cells, based on tyrosine phosphorylation and translocation into lipid rafts, sIgM signaling was shown to be enhanced by Zap70. The different consequences of signaling through sIgM were associated with biological prognosis indicators. These included high levels of CD38, lack of mutations in the IgVH chain genes, preferential usage of full-length CD79b, and severe clinical stage. Thus, modification of sIgM-induced signaling could be a therapeutic approach. PMID- 15749914 TI - Alloreactive T cell responses and acute rejection of single class II MHC disparate heart allografts are under strict regulation by CD4+ CD25+ T cells. AB - Skin but not vascularized cardiac allografts from B6.H-2bm12 mice are acutely rejected by C57BL/6 recipients in response to the single class II MHC disparity. The underlying mechanisms preventing acute rejection of B6.H-2bm12 heart allografts by C57BL/6 recipients were investigated. B6.H-2bm12 heart allografts induced low levels of alloreactive effector T cell priming in C57BL/6 recipients, and this priming was accompanied by low-level cellular infiltration into the allograft that quickly resolved. Recipients with long-term-surviving heart allografts were unable to reject B6.H-2bm12 skin allografts, suggesting potential down-regulatory mechanisms induced by the cardiac allografts. Depletion of CD25+ cells from C57BL/6 recipients resulted in 15-fold increases in alloreactive T cell priming and in acute rejection of B6.H-2bm12 heart grafts. Similarly, reconstitution of B6.Rag(-/-) recipients with wild-type C57BL/6 splenocytes resulted in acute rejection of B6.H-2bm12 heart grafts only if CD25+ cells were depleted. These results indicate that acute rejection of single class II MHC disparate B6.H-2bm12 heart allografts by C57BL/6 recipients is inhibited by the emergence of CD25+ regulatory cells that restrict the clonal expansion of alloreactive T cells. PMID- 15749916 TI - Differential regulation of granzyme and perforin in effector and memory T cells following smallpox immunization. AB - Primary immunization of healthy adults with vaccinia virus induces a local vesicle or "take" in the majority of vaccinees that previously has been shown to correlate with protection against smallpox. However, the immunologic mechanisms underlying this protective response in humans are not well characterized. We have studied human CD8+ T cells for the expression patterns of phenotypic markers and cytolytic effector molecules before and after primary smallpox immunization using nine-color polychromatic flow cytometry. One month after immunization, vaccinees developed vaccinia virus-specific CD8+ T cells with an effector cell phenotype containing both granzyme A and granzyme B. One year after immunization, we found a significant decrease in granzyme B containing cells and an increased memory cell phenotype in virus-specific CD8+ T cells. Perforin was rarely expressed directly ex vivo, but was highly expressed after Ag-specific activation in vitro. Together, these data suggest an important role for effector CD8+ T cells in controlling poxvirus infection, and have implications for our understanding of human CD8+ T cell differentiation. PMID- 15749917 TI - Prolonged lymphopenia, lymphoid depletion, and hypoprolactinemia in children with nosocomial sepsis and multiple organ failure. AB - Lymphopenia and lymphoid depletion occur in adults dying of sepsis. Prolactin increases Bcl-2 expression, suppresses stress-induced lymphocyte apoptosis, and improves survival from experimental sepsis. We hypothesized that prolonged lymphopenia, lymphoid depletion, and hypoprolactinemia occur in children dying with sepsis and multiple organ failure (MOF). Fifty-eight critically ill children with and 55 without MOF admitted to a university hospital pediatric intensive care unit were enrolled in a prospective, longitudinal, observational clinical study. Prolactin levels and absolute lymphocyte count were measured on days 1, 3, 7, 14, and 21. Lymph node, thymus, and spleen autopsy specimens were examined for lymphoid depletion, with immunohistochemical staining for CD4, CD20, and CD21 and for lymphoid apoptosis. Prolonged lymphopenia (absolute lymphocyte count < 1000 for >7 days) occurred only in children with MOF (29 vs 0%, p < 0.05) and was associated independently with nosocomial infection (odds ratio (OR), 5.5, 95% confidence interval (CI), 1.7-17, p < 0.05), death (OR, 6.8, 95% CI, 1.3-34, p < 0.05), and splenic and lymph node hypocellularity (OR, 42, 95% CI, 3.7-473, p < 0.05). Lymphocyte apoptosis and ante/postmortem infection were observed only in children with lymphoid depletion. Prolonged hypoprolactinemia (>7 days) was more common in children with MOF (17 vs 2%, p < 0.05) and was associated independently with prolonged lymphopenia (OR, 8.3, 95% CI, 2.1-33, p < 0.05) and lymphoid depletion (OR, 12.2, 95% CI, 2.2-65, p < 0.05). Prolonged lymphopenia and apoptosis-associated depletion of lymphoid organs play a role in nosocomial sepsis-related death in critically ill children. Prolonged hypoprolactinemia is a previously unrecognized risk factor for this syndrome. PMID- 15749918 TI - CD1a and CD1c activate intrathyroidal T cells during Graves' disease and Hashimoto's thyroiditis. AB - Molecular studies have shown that CD1 proteins present self and foreign lipid Ags to T cells, but the possible roles of CD1 in human autoimmune diseases in vivo are not known, especially for the group 1 CD1 isoforms (CD1a, CD1b, and CD1c). To investigate the hypothesis that CD1-restricted T cells might be activated and home to target tissues involved in Hashimoto's thyroiditis and Graves' disease, we performed ex vivo analysis of lymphocytes from peripheral blood and autoinflammatory lesions of thyroid tissue. Immunofluorescence analysis identified two types of CD1-expressing APCs in inflamed thyroid tissues. CD1a, CD1b, and CD1c were expressed on CD83+ dendritic cells, and CD1c was expressed on an abundant population of CD20+ IgD+ CD23- CD38- B cells that selectively localized to the mantle zone of lymphoid follicles within the thyroid gland. CD1c restricted, glycolipid-specific T cells could not be detected in the peripheral blood, but were present in polyclonal lymphocyte populations isolated from affected thyroid glands. In addition, polyclonal thyroid-derived lymphocytes and short-term T cell lines were found to recognize and lyse targets in a CD1a- or CD1c-dependent manner. The targeting of CD1-restricted T cells and large numbers of CD1-expressing APCs to the thyroid gland during the early stages of autoimmune thyroiditis suggests a possible effector function of CD1-restricted T cells in tissue destruction and point to a new model of organ-specific autoimmune disease involving lipid Ag presentation. PMID- 15749919 TI - Antigen-specific B cells are required as APCs and autoantibody-producing cells for induction of severe autoimmune arthritis. AB - B cells play an important role in rheumatoid arthritis, but whether they are required as autoantibody-producing cells as well as APCs has not been determined. We assessed B cell autoantibody and APC functions in a murine model of autoimmune arthritis, proteoglycan (PG)-induced arthritis, using both B cell-deficient mice and Ig-deficient mice (mIgM) mice that express an H chain transgene encoding for membrane-bound, but not secreted, IgM. The IgH transgene, when paired with endogenous lambda L chain, recognizes the hapten 4-hydroxy-3-nitro-phenyl acetyl and is expressed on 1-4% of B cells. B cell-deficient and mIgM mice do not develop arthritis after immunization with PG. In adoptive transfer of PG-induced arthritis into SCID mice, T cells from mIgM mice immunized with PG were unable to transfer disease even when B cells from PG-immunized wild-type mice were provided, suggesting that the T cells were not adequately primed and that Ag specific B cells may be required. In fact, when PG was directly targeted to the B cell Ig receptor through a conjugate of 4-hydroxy-3-nitrophenyl acetyl-PG, T cells in mIgM mice were activated and competent to transfer arthritis. Such T cells caused mild arthritis in the absence of autoantibody, demonstrating a direct pathogenic role for T cells activated by Ag-specific B cells. Transfer of arthritic serum alone induced only mild and transient arthritis. However, both autoreactive T cells and autoantibody are required to cause severe arthritis, indicating that both B cell-mediated effector pathways contribute synergistically to autoimmune disease. PMID- 15749920 TI - IL-10-dependent suppression of experimental allergic encephalomyelitis by Th2 differentiated, anti-TCR redirected T lymphocytes. AB - We previously showed that transgenically expressed chimeric Ag-MHC-zeta receptors can Ag-specifically redirect T cells against other T cells. When the receptor's extracellular Ag-MHC domain engages cognate TCR on an Ag-specific T cell, its cytoplasmic zeta-chain stimulates the chimeric receptor-modified T cell (RMTC). This induces effector functions such as cytolysis and cytokine release. RMTC expressing a myelin basic protein (MBP) 89-101-IAs-zeta receptor can be used therapeutically, Ag-specifically treating murine experimental allergic encephalomyelitis (EAE) mediated by MBP89-101-specific T cells. In initial studies, isolated CD8+ RMTC were therapeutically effective whereas CD4+ RMTC were not. We re-examine here the therapeutic potential of CD4+ RMTC. We demonstrate that Th2-differentiated, though not Th1-differentiated, CD4+ MBP89-101-IAs-zeta RMTC prevent actively induced or adoptively transferred EAE, and treat EAE even after antigenic diversification of the pathologic T cell response. The Th2 RMTC both Th2-deviate autoreactive T cells and suppress autoantigen-specific T cell proliferation. IL-10 is critical for the suppressive effects. Anti-IL-10R blocks RMTC-mediated modulation of EAE and suppression of autoantigen proliferation, as well as the induction of IL-10 production by autoreactive T cells. In contrast to IL-10, IL-4 is required for IL-4 production by, and hence Th2 deviation of autoreactive T cells, but not the therapeutic activity of the RMTC. These results therefore demonstrate a novel immunotherapeutic approach for the Ag-specific treatment of autoimmune disease with RMTC. They further identify an essential role for IL-10, rather than Th2-deviation itself, in the therapeutic effectiveness of these redirected Th2 T cells. PMID- 15749921 TI - Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8+ and CD4+ T cell responses in patients with metastatic prostate cancer. AB - Telomerase reverse transcriptase (hTERT) represents an attractive target for cancer immunotherapy because hTERT is reactivated in most human tumors. A clinical trial was initiated in which hTERT mRNA-transfected dendritic cells (DC) were administered to 20 patients with metastatic prostate cancer. Nine of these subjects received DC transfected with mRNA encoding a chimeric lysosome associated membrane protein-1 (LAMP) hTERT protein, allowing for concomitant induction of hTERT-specific CD8+ and CD4+ T cell responses. Treatment was well tolerated. Intense infiltrates of hTERT-specific T cells were noted at intradermal injection sites after repeated vaccination. In 19 of 20 subjects, expansion of hTERT-specific CD8+ T cells was measured in the peripheral blood of study subjects, with 0.9-1.8% of CD8+ T cells exhibiting Ag specificity. Patients immunized with the chimeric LAMP hTERT vaccine developed significantly higher frequencies of hTERT-specific CD4+ T cells than subjects receiving DC transfected with the unmodified hTERT template. Moreover, CTL-mediated killing of hTERT targets was enhanced in the LAMP hTERT group, suggesting that an improved CD4+ response could augment a CTL response. Vaccination was further associated with a reduction of prostate-specific Ag velocity and molecular clearance of circulating micrometastases. Our findings provide a rationale for further development of hTERT-transfected DC vaccines in the treatment of prostate and other cancers. PMID- 15749922 TI - Immunization with lentiviral vector-transduced dendritic cells induces strong and long-lasting T cell responses and therapeutic immunity. AB - Dendritic cell (DC) therapies are currently being evaluated for the treatment of cancer. The majority of ongoing clinical trials use DCs loaded with defined antigenic peptides or proteins, or tumor-derived products, such as lysates or apoptotic cells, as sources of Ag. Although several theoretical considerations suggest that DCs expressing transgenic protein Ags may be more effective immunogens than protein-loaded cells, methods for efficiently transfecting DCs are only now being developed. In this study we directly compare the immunogenicity of peptide/protein-pulsed DCs with lentiviral vector-transduced DCs, and their comparative efficacy in tumor immunotherapy. Maturing, bone marrow derived DCs can be efficiently transduced with lentiviral vectors, and transduction does not affect DC maturation, plasticity, or Ag presentation function. Transduced DCs efficiently process and present both MHC class I- and II restricted epitopes from the expressed transgenic Ag OVA. Compared with peptide- or protein-pulsed DCs, lentiviral vector-transduced DCs elicit stronger and longer-lasting T cell responses in vivo, as measured by both in vivo killing assays and intracellular production of IFN-gamma by Ag-specific T cells. In the B16-OVA tumor therapy model, the growth of established tumors was significantly inhibited by a single immunization using lentiviral vector-transduced DCs, resulting in significantly longer survival of immunized animals. These results suggest that compared with Ag-pulsed DCs, vaccination with lentiviral vector transduced DCs may achieve more potent antitumor immunity. These data support the further development of lentiviral vectors to transduce DCs with genes encoding Ags or immunomodulatory adjuvants to generate and control systemic immune responses. PMID- 15749924 TI - High-efficiency RNA interference in human embryonic stem cells. AB - RNA interference methodology suppresses gene expression, thus mimicking loss-of function mutation and enabling in vitro and in vivo gene function analysis. In this study, we used retroviral and lentiviral vectors to deliver small interfering RNAs and report high-efficiency silencing of a green fluorescent protein (GFP) trans gene and the stem cell-specific transcription factors Oct4/POU5F1 and Nanog in human embryonic stem cells. Gene knockdown of Oct4 and Nanog promotes differentiation, thereby demonstrating a role for these factors in human embryonic stem cell self-renewal. PMID- 15749925 TI - An autogeneic feeder cell system that efficiently supports growth of undifferentiated human embryonic stem cells. AB - Human embryonic stem cells (hESCs) have great potential as a source of cells for therapeutic uses, but their culture requires the support of mouse or human cells, either directly as a feeder cell layer or indirectly as a source of conditioned medium in feeder-free culture systems. Unfortunately, the risks of cross-transfer of pathogens from xenogeneic or allogeneic feeders or cell by-products limit their medical applications. In addition, not all human feeders support the growth of hESCs equally well, and ethical concerns have been raised regarding the derivation of feeder cells from aborted human fetuses. We report here the culture of hESCs on a novel feeder cell system, comprising fibroblast-like cells derived from the spontaneous differentiation of hESCs. Isogenicity of the hESCs and hESC derived fibroblasts was confirmed by micro satellite analysis. The nature of the hESC-derived fibroblasts was identified by the expression of specific markers. This feeder system permits continuous growth of undifferentiated and pluripotent hESCs, as demonstrated by the expression of specific hESC markers, by the formation of teratomas after injection of hESCs into severely combined immunodeficient mice, and by in vitro differentiation of hESCs into differentiated cells of ectodermal, endodermal, and mesodermal origin. Feeder cells derived from hESCs offers a potentially more secure autogeneic and genotypically homogenous system for the growth of undifferentiated hESCs. PMID- 15749927 TI - A modified cord blood collection method achieves sufficient cell levels for transplantation in most adult patients. AB - Umbilical cord blood transplantation (UCBT) has been used increasingly in both pediatric and adult patients. The total nucleated cell (NC) dose infused is the most critical factor in determining speed of engraftment and survival. Using standard collection techniques, the mean NC content of UCB units is about 10 x 10(8) and only 25% of these units reach the target cell dose of 2 x 10(7)/kg in UCBT patients weighing 50-70 kg. We have designed a modified placental/umbilical two-step collection method in which a standard blood fraction obtained by umbilical venipuncture is combined with a second fraction harvested after placental perfusion with 50 ml heparinized 0.9% saline. This second fraction contributed 32% volume and 15% NCs to the whole UCB unit (123.7 +/- 50.1 ml and 1.26 +/- 0.52 x 10(9) NC). The proportion of progenitor cells in both fractions was not significantly different, indicating that the hematopoietic potential of these larger units is 20% (range, 2%-100%) higher than UCB units collected by standard methods. In addition, the bacterial contamination rate associated with this novel collection method (2.78%) compares favorably. Since 1998 we have further enriched our units by processing only UCB units over 0.8 x 10(9) NCs, resulting in a 36% cell increment (1.46 +/- 0.52 x 10(9) NCs). Thus, 84% and 54% of the Madrid UCB Bank inventory would fulfill the target cell dose of 2 x 10(7)/kg in patients weighing 50 and 65 kg, respectively. This significant UCB banking improvement gives larger pediatric and adult patients a greater chance of finding adequate grafts in order to achieve better clinical outcomes after UCBT. PMID- 15749926 TI - Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. AB - Previous studies have shown that prolonged propagation of undifferentiated human embryonic stem cells (hESCs) requires conditioned medium from mouse embryonic feeders (MEF-CM) as well as matrix components. Because hESCs express growth factor receptors, including those for basic fibroblast growth factor (bFGF), stem cell factor (SCF), and fetal liver tyrosine kinase-3 ligand (Flt3L), we evaluated these and other growth factors for their ability to maintain undifferentiated hESCs in the absence of conditioned medium. We found cultures maintained in bFGF alone or in combination with other factors showed characteristics similar to MEF CM control cultures, including morphology, surface marker and transcription factor expression, telomerase activity, differentiation, and karyotypic stability. In contrast, cells in media containing Flt-3L, thrombopoietin, and SCF, individually or in combination, showed almost complete differentiation after 6 weeks in culture. These data demonstrate that hESCs can be maintained in nonconditioned medium using growth factors. PMID- 15749928 TI - Rac2-deficient hematopoietic stem cells show defective interaction with the hematopoietic microenvironment and long-term engraftment failure. AB - The hematopoietic-specific Rho GTPase, Rac2, regulates a variety of cellular functions including cell shape changes, motility, integrin-dependent adhesion, and apoptosis. In the study reported here, we demonstrate that wild-type (WT) hematopoietic stem cells/progenitors (HSC/P) preferentially engraft in nonablated Rac2(-/-) bone marrow. In addition, primitive Rac2(-/-) HSC/P transplanted into lethally irradiated WT recipients showed a significant competitive defect compared with WT cells. These defects appeared to be related to HSC/P-intrinsic defective microenvironment interactions, since Rac2(-/-) cells showed less adhesion to the femur bone marrow density 1 (FBMD-1) stromal cell line, a lower frequency of cobblestone area-forming cells, and lower performance in long-term marrow cultures in vitro when compared with WT cells. In contrast, primitive Rac2(-/-) hematopoietic cells exhibited normal progenitor colony formation in semisolid medium in vitro and normal proliferation in the steady state in vivo when compared with WT cells. Taken together, these data suggest that Rac2(-/-) stem/progenitor cells exhibit abnormal interaction with the hematopoietic microenvironment, which leads to defective long-term engraftment. PMID- 15749929 TI - Macrophage colony-stimulating factor (M-CSF), as well as granulocyte colony stimulating factor (G-CSF), accelerates neovascularization. AB - It has been reported that bone marrow cells (BMCs) differentiate into endothelial cells of blood vessels, and that granulocyte colony-stimulating factor (G-CSF) mobilizes progenitors in the BMCs to the peripheral blood, while macrophage colony-stimulating factor (M-CSF) augments the production of monocytes. We examined whether M-CSF augments the differentiation of BMCs into endothelial cells of blood vessels using a hindlimb-ischemic model. Either G-CSF or M-CSF, or both, was administered to the hindlimb-ischemic mice for 3 days. Both M-CSF and G CSF augmented the differentiation of BMCs into endothelial cells of blood vessels through vascular endothelial cell growth factor (VEGF), resulting in early recovery of blood flow in the ischemic limbs. PMID- 15749930 TI - Repair of infarcted myocardium mediated by transplanted bone marrow-derived CD34+ stem cells in a nonhuman primate model. AB - Rodent and human clinical studies have shown that transplantation of bone marrow stem cells to the ischemic myocardium results in improved cardiac function. In this study, cynomolgus monkey acute myocardial infarction was generated by ligating the left anterior descending artery, and autologous CD34(+) cells were transplanted to the peri-ischemic zone. To track the in vivo fate of transplanted cells, CD34(+) cells were genetically marked with green fluorescent protein (GFP) using a lentivirus vector before transplantation (marking efficiency, 41% on average). The group receiving cells (n = 4) demonstrated improved regional blood flow and cardiac function compared with the saline-treated group (n =4) at 2 weeks after transplant. However, very few transplanted cell-derived, GFP-positive cells were found incorporated into the vascular structure, and GFP-positive cardiomyocytes were not detected in the repaired tissue. On the other hand, cultured CD34(+) cells were found to secrete vascular endothelial growth factor (VEGF), and the in vivo regional VEGF levels showed a significant increase after the transplantation. These results suggest that the improvement is not the result of generation of transplanted cell-derived endothelial cells or cardiomyocytes; and raise the possibility that angiogenic cytokines secreted from transplanted cells potentiate angiogenic activity of endogenous cells. PMID- 15749931 TI - A new strategy for treatment of malignant tumor: intra-bone marrow-bone marrow transplantation plus CD4- donor lymphocyte infusion. AB - Donor lymphocyte infusion (DLI) is clinically used for the treatment of malignant tumors. We have found recently that intra-bone marrow-bone marrow transplantation (IBM-BMT) can be used to treat various autoimmune diseases, even when radiation doses are reduced. In addition, recently we have found that IBM-BMT can prevent not only graft failure but also graft-versus-host disease (GvHD). Based on these findings, we attempted to prevent and treat the progression of a tumor (Meth-A cell line: BALB/c-derived fibrosarcoma) by DLI plus IBM-BMT. When the tumors had grown to approximately 10 x 10 mm, the tumor-bearing BALB/c (H-2(d)) mice were irradiated with 5 Gy, and whole spleen cells from C57BL/6J (B6) (H-2(b)) mice (as DLI) were then intravenously injected into the BALB/c mice. Simultaneously, bone marrow cells (BMCs) from B6 mice were injected directly into the bone marrow cavity of the BALB/c mice (IBM-BMT). The tumors decreased in size, but the mice died of GvHD. However, when CD4(+) T-cell-depleted spleen cells were used for DLI, the recipients showed only mild GvHD and survived longer, due to the slow growth of the tumor. In contrast, when CD8(+) T-cell-depleted spleen cells were used for DLI, the recipients showed more severe GvHD than those injected with whole spleen cells. These results suggest that IBM-BMT plus DLI (the depletion or reduction of a certain cell population like CD4(+) T cells) could be helpful to suppress both GvHD and tumor growth. PMID- 15749932 TI - Characterization of ionic currents in human mesenchymal stem cells from bone marrow. AB - This study characterized functional ion channels in cultured undifferentiated human mesenchymal stem cells (hMSCs) from bone marrow with whole-cell patch clamp and reverse transcription polymerase chain reaction (RT-PCR) techniques. Three types of outward currents were found in hMSCs, including a noise-like rapidly activating outward current inhibited by the large conductance Ca(2+)-activated K(+) channel (I(KCa)) blocker iberiotoxin, a transient outward K(+) current (I(to)) suppressed by 4-aminopyridine (4-AP), and a delayed rectifier K(+) current (IK(DR))-like ether-a-go-go (eag) K(+) channel. In addition, tetrodotoxin sensitive sodium current (I(Na.TTX)) and nifedipine-sensitive L-type Ca(2+) current (I(Ca.L)) were also detected in 29% and 15% hMSCs, respectively. Moreover, RT-PCR revealed the molecular evidence of high levels of mRNA for the functional ionic currents, including human MaxiK for I(KCa), Kv4.2 and Kv1.4 for I(to), heag1 for IK(DR), hNE-Na for I(Na.TTX), and CACNAIC for I(Ca.L). These results demonstrate that multiple functional ion channel currents--that is, I(KCa), I(to), heag1, I(Na.TTX), and I(Ca.L)--are expressed in hMSCs from bone marrow. PMID- 15749933 TI - Neurons derived from human mesenchymal stem cells show synaptic transmission and can be induced to produce the neurotransmitter substance P by interleukin-1 alpha. AB - Mesenchymal stem cells (MSCs) exhibit immune-suppressive properties, follow a pattern of multilineage differentiation, and exhibit transdifferentiation potential. Ease in expansion from adult bone marrow, as well as its separation from ethical issues, makes MSCs appealing for clinical application. MSCs treated with retinoic acid resulted in synaptic transmission, based on immunostaining of synaptophysin and electrophysiological studies. In situ hybridization indicated that the neurotransmitter gene preprotachykinin-I was expressed in these cells. However, translation of this gene only occurred after stimulation with interleukin (IL)-1 alpha. This effect was blunted by costimulation with IL-1 receptor antagonist. This study reports on the ability of MSCs to be transdifferentiated into neurons with functional synapses with the potential to become polarized towards producing specific neurotransmitters. PMID- 15749934 TI - Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. AB - Bone marrow mesenchymal stem cells (MSCs) can differentiate into several types of mesenchymal cells, including osteocytes, chondrocytes, and adipocytes, but, under appropriate experimental conditions, can also differentiate into nonmesenchymal cells--for instance, neural cells. These observations have raised interest in the possible use of MSCs in cell therapy strategies for various neurological disorders. In the study reported here, we addressed the question of in vitro differentiation of MSCs into functional neurons. First, we demonstrate that when they are co-cultured with cerebellar granule neurons, adult MSCs can express neuronal markers. Two factors are needed for the emergence of neuronal differentiation of the MSCs: the first one is nestin expression by MSCs (nestin is a marker for the responsive character of MSCs to extrinsic signals), and the second one is a direct cell-cell interaction between neural cells and MSCs that allows the integration of these extrinsic signals. Three different approaches suggest that neural phenotypes arise from MSCs by a differentiation rather than a cell fusion process, although this last phenomenon can also coexist. The expression of several genes--including sox, pax, notch, delta, frizzled, and erbB -was analyzed by quantitative reverse transcription polymerase chain reaction (RT PCR) in order to further characterize the nestin-positive phenotype compared to the nestin-negative one. An overexpression of sox2, sox10, pax6, fzd, erbB2, and erbB4 is found in nestin-positive MSCs. Finally, electrophysiological analyses demonstrate that MSC-derived neuron-like cells can fire single-action potentials and respond to several neurotransmitters such as GABA, glycine, and glutamate. We conclude that nestin-positive MSCs can differentiate in vitro into excitable neuron-like cells. PMID- 15749936 TI - Cell surface and transcriptional characterization of human adipose-derived adherent stromal (hADAS) cells. AB - Adult human subcutaneous adipose tissue contains cells with intriguing multilineage developmental plasticity, much like marrow-derived mesenchymal stem cells. Putative stem or progenitor cells from fat have been given many different names in the literature, reflecting an early and evolving consensus regarding their phenotypic characterization. The study reported here used microarrays to evaluate over 170 genes relating to angiogenesis and extracellular matrix in undifferentiated, early-passage human adipose-derived adherent stromal (hADAS) cells isolated from three separate donors. The hADAS populations unanimously transcribed 66% of the screened genes, and 83% were transcribed by at least two of the three populations. The most highly transcribed genes relate to functional groupings such as cell adhesion, matrix proteins, growth factors and receptors, and proteases. The transcriptome of hADAS cells demonstrated by this work reveals many similarities to published profiles of bone marrow mesenchymal stem cells (MSCs). In addition, flow analysis of over 24 hADAS cell surface proteins (n = 7 donors) both confirms and expands on the existing literature and reveals strong intergroup correlation, despite an inconsistent nomenclature and the lack of standardized protocols for cell isolation and culture. Finally, based on flow analysis and reverse transcription polymerase chain reaction studies, our results suggest that hADAS cells do not express several proteins that are implicated as markers of "stemness" in other stem cell populations, including telomerase, CD133, and the membrane transporter ABCG2. PMID- 15749935 TI - Induction of intervertebral disc-like cells from adult mesenchymal stem cells. AB - The potential of adult mesenchymal stem cells (MSCs) to differentiate towards cartilage, bone, adipose tissue, or muscle is well established. However, the capacity of MSCs to differentiate towards intervertebral disc (IVD)-like cells is unknown. The aim of this study was to compare the molecular phenotype of human IVD cells and articular chondrocytes and to analyze whether mesenchymal stem cells can differentiate towards both cell types after transforming growth factor beta (TGF beta)-mediated induction in vitro. Bone marrow-derived MSCs were differentiated in spheroid culture towards the chondrogenic lineage in the presence of TGF beta(3) dexamethasone, and ascorbate. A customized cDNA-array comprising 45 cartilage-, bone-, and stem cell-relevant genes was used to quantify gene expression profiles. After TGF beta-mediated differentiation, MSC spheroids turned positive for collagen type II protein and expressed a large panel of genes characteristic for chondrocytes, including aggrecan, decorin, fibromodulin, and cartilage oligomeric matrix protein, although at levels closer to IVD tissue than to hyaline articular cartilage. Like IVD tissue, the spheroids were strongly positive for collagen type I and osteopontin. MSC spheroids expressed more differentiation markers at higher levels than culture-expanded IVD cells and chondrocytes, which both dedifferentiated in monolayer culture. In conclusion, mesenchymal stem cells adopted a gene expression profile that resembled native IVD tissue more closely than native joint cartilage. Thus, these cells may represent an attractive source from which to obtain IVD-like cells, whereas modification of culture conditions is required to approach the molecular phenotype of chondrocytes in hyaline cartilage. PMID- 15749937 TI - Effects of ciliary neurotrophic factor on differentiation of late retinal progenitor cells. AB - Ciliary neurotrophic factor (CNTF) has been shown to be a potent regulator of retinal cell differentiation. The present study was undertaken to investigate the effects of CNTF on in vitro differentiation of expanded late retinal progenitor cells. Retinal progenitor cells used in these studies were isolated from the neural retina of postnatal day-1 green fluorescent protein (GFP) transgenic mice. The resulting GFP-positive neurospheres were dissociated into a single-cell suspension and grown on poly-D-lysine/laminin-coated tissue culture flasks or slides to generate adherent retinal progenitor cells. These adherent cells were treated with 20 ng/ml of CNTF for up to 14 days, and expression of specific retinal cell markers was determined by immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR), and immunoblot analysis. In vitro studies showed that CNTF treatment of late retinal progenitor cells resulted in changes in cellular morphology. Immunocytochemical studies showed an increase in the proportion of cells expressing markers of bipolar cells but not rod differentiation. In addition, an increase in the proportion of cells expressing glial cell markers was observed. RT-PCR analysis showed downregulation in Hes1, Nestin, Notch1, and Pax6 transcripts along with a concomitant increase in protein kinase C (PKC) alpha and glial fibrillary acidic protein (GFAP) transcripts. These findings were confirmed by immunoblot analysis, where downregulation in Nestin expression and simultaneous upregulation in PKC alpha and GFAP were observed. The data indicate that CNTF treatment of multipotential late retinal progenitors increases the proportion of cells that express markers of bipolar neurons and glia. PMID- 15749938 TI - Retinoic acid stimulates the dynamics of mouse gastric epithelial progenitors. AB - The gastric epithelial progenitors proliferate and undergo bipolar migration associated with their differentiation into pit, parietal, and zymogenic cell lineages. Retinoids have long been known to modulate proliferation and differentiation of various renewing epithelia, and the expression of their receptors has been demonstrated in the gastric mucosa. The aim of this study was to examine the effects of retinoic acid on progenitor cell proliferation and cell lineage formation in the mouse stomach. By using subcutaneously inserted osmotic pumps, mice were continuously infused with all-trans retinoic acid (5 mg/kg per day) for 3 days. To label S-phase cells and their progeny, bromodeoxyuridine was administered for different time intervals. Analysis of gastric mucosal tissues of retinoic acid-treated mice revealed a significant increase in the number of S phase progenitor cells and an enhancement in the production of their progeny. The life span of pit cells was reduced, and their apoptosis became apparent at the luminal surface. Immunofluoresence probing of pit, parietal and enteroendocrine cell lineages in control and retinoic acid-treated mice showed no significant change in their labeling pattern. However, there was an increase in the labeled gland area of zymogenic cells. In conclusion, 3-day treatment of retinoic acid enhances the proliferation of gastric epithelial progenitors and the dynamics of their progeny. PMID- 15749940 TI - The molecular perspective: major histocompatibility complex. PMID- 15749941 TI - Hypertension in a dental school patient population. PMID- 15749939 TI - Induction of oligodendrocytes from adult human olfactory epithelial-derived progenitors by transcription factors. AB - Neurosphere-forming cell (NSFC) lines have been derived from cultures of adult olfactory neuroepithelium obtained from patients and cadavers. These progenitors remain undifferentiated when maintained in minimal essential medium with 10% fetal bovine serum, but have the potential to differentiate along glial or neuronal lineages. However, few of these cells ever express mature neuronal or glial markers in defined medium. To evaluate the potential of NSFCs to form oligodendrocytes, two transcription factors, Olig2 and Nkx2.2, were introduced into NSFCs to determine whether their expression is sufficient for oligodendrocyte differentiation, as has been shown in the embryonic avian and murine central nervous systems in vivo. NSFCs transfected with Olig2 or Nkx2.2 alone exhibited no phenotypic lineage restriction. In contrast, simultaneous transfection of Olig2 and Nkx2.2 cDNA produced characteristic oligodendrocyte morphology and antigenicity, including myelin basic protein (MBP). Furthermore, a population of Olig2-expressing NSFCs also expressed Sox10. Cotransfection of NSFCs with Nkx2.2 and Sox10, but not Olig2 and Sox10, produced a MBP(+) oligodendrocytic phenotype. Coculture of NSFCs transfected with Olig2 and Nkx2.2 or Nkx2.2 and Sox10 with purified sensory neurons, demonstrated frequent contacts between NSFC processes and axons, including the early stages of ensheathment. These studies demonstrate transcription factors governing early development of chick and mouse oligodendrocyte formation, also apply to human progenitors isolated from adult olfactory neuroepithelium. Our long-term goal is to develop cell populations for future studies used to determine the therapeutic utility of these olfactory-derived NSFCs for autologous transplantation into donors with central nervous system trauma or neurodegenerative diseases. PMID- 15749942 TI - Issues and challenges in special care dentistry. PMID- 15749943 TI - Variation in periodontal diagnosis and treatment planning among clinical instructors. AB - Consistency in clinical decision making may be necessary for reliable assessment of student performance and teaching effectiveness, yet little has been done to examine variation in periodontal diagnosis and treatment planning among dental school faculty. The purpose of this investigation was to examine variation among faculty in diagnosis and management of common periodontal diseases. Twenty-seven clinical instructors (periodontists, general dentists, dental hygienists, and first- and second-year periodontal graduate students) reviewed three web-based cases and answered a brief questionnaire focusing on radiographic interpretation, periodontal diagnosis, and treatment planning. Response rates for the three cases ranged from 62 percent to 70 percent. Clinical instructors' rating of percent bone loss in the majority of cases varied between three descriptive categories for the same tooth. Greater consistency in periodontal diagnosis was noted within the graduate student group as compared to periodontal and dental hygiene faculty groups. Diagnoses offered for one of the three patients varied between gingivitis and chronic and aggressive periodontitis. Six to nineteen different treatment plans (many with subtle differences) were submitted for each of the three cases. Inter-rater variation was qualitatively more prevalent than intra-rater variation. To our knowledge, this is the first study to document substantial variation among instructors in radiographic interpretation, diagnosis, and treatment planning for common periodontal diseases. Qualitative judgments speculating on the impact of variability among dental school faculty on student performance and patient care can be made but as yet remain unknown. Consistent use of accepted practice guidelines and greater consensus-building opportunities may decrease variation among faculty and enhance dental education. PMID- 15749944 TI - Measurement of changes in empathy during dental school. AB - Empathy in the health care setting is the ability to understand a patient's experiences and feelings and the capability to communicate this understanding. Although empathy has been shown to play an important role in the dentist-patient relationship and is a core competence for dentists, no measure of empathy has been validated in the dental setting. Further, little is known about changes in empathy during dental school. We examined the psychometric properties of a measure of empathy applied to the dental school setting and compared levels of empathy in dental students across their four years of training. One hundred and thirty students completed a survey including the Jefferson Scale of Physician Empathy (JSPE). The JSPE was found to be both a reliable and valid empathy measure for dental students with similar psychometric properties to those found in medical students, residents, and physicians. Further, first-year dental students had significantly higher empathy scores than students in any subsequent year. Consistent with the literature in medical settings, the timing of the decline in empathy levels corresponded to increases in patient exposure. We suggest that training students in the interpersonal skills designed to enhance the dentist-patient relationship should continue throughout dental school training. PMID- 15749945 TI - Knowledge of oral and physical manifestations of anorexia and bulimia nervosa among dentists and dental hygienists. AB - Despite the crucial role oral health care providers can have in the early identification of eating disorders and the referral and case management of patients with these disorders, little is known concerning their knowledge of oral complications of these disorders. The purpose of this study was to determine the knowledge among dentists and dental hygienists concerning the oral and physical manifestations of eating disorders. Employing a randomized cross-sectional study, data were collected from 576 dentists and dental hygienists randomly selected from the American Dental Association and the American Dental Hygienists' Association. Results indicated low scores concerning knowledge of oral cues, physical cues of anorexia, and physical cues of bulimia among study participants. More dental hygienists than dentists correctly identified oral manifestations of eating disorders (p=.001) and physical cues of anorexia (p=.010) and bulimia (p=.002). As the first health professional to identify oral symptoms of eating disorders, the most important task of the dental care provider when identifying oro-dental signs of eating disorders is to ensure that the patient receives treatment. Implications for education include the addition of conceptual, procedural, and skill-based curricula objectives addressing etiologic assessment and patient communication--thus increasing behavioral capacity for delivery of restorative care and patient referral. PMID- 15749946 TI - What determines positive student perceptions of extramural clinical rotations? An analysis using 2003 ADEA Senior Survey data. AB - Extramural clinical rotations are an integral part of many dental school curricula. Schools in The Robert Wood Johnson Foundation/The California Endowment Pipeline, Profession, and Practice program are increasing student extramural opportunities to expose students to patients of different needs, cultures, and dental delivery modes. Using data from the American Dental Education Association (ADEA) 2003 Senior Survey, the Pipeline, Profession, and Practice National Evaluation Team studied graduating dental students' perceptions about their extramural rotations. This analysis was designed to determine the factors associated with students' perception of their extramural clinical rotations: was it a positive experience in their dental education, and did it improve students' perceptions of their ability to provide care for racially, ethnically, and culturally diverse groups? The respondents were 2,950 graduating seniors who reported at least one week of extramural rotation experience. After controlling for both individual and school level characteristics, this study found that race/ethnicity, a stronger socially conscious attitudes score, number of weeks spent in extramural rotations, and the student's rating of time spent in extramural rotations were significant determinants of the extramural rotations being reported as positive experiences. With respect to improving students' ability to provide care to racially, ethnically, and culturally diverse groups, more positive student socially conscious attitudes, a greater number of weeks spent in the rotations, a stronger service orientation for selecting dentistry as a career, and the students' rating of their time spent in extramural rotations were significant determinants. In conclusion, there is some evidence that time spent in extramural rotations may be perceived as positive dental school experiences and, for some students, may prepare them to work effectively with culturally diverse patients by the time of dental school graduation. PMID- 15749947 TI - Predictive validity of dental hygiene competency assessment measures on one-shot clinical licensure examinations. AB - The purpose of this study was to examine the predictive validity of traditional and nontraditional dental hygiene competency assessment measures on one-shot clinical licensure examinations in a baccalaureate dental hygiene program. Traditional assessment data including overall grade point average (GPA), Clinical GPA, National Board Dental Hygiene Examination (NBDHE) scores, and Central Regional Dental Testing Service (CRDTS) scores along with nontraditional assessment data in the form of Portfolio scores were collected from seventy-four students. Factor analysis and subsequent linear regression modeling were used to explore the ability of four variables (Overall GPA, NBDHE, Portfolios, and Clinical GPA) to predict one-shot clinical licensure examination (CRDTS) scores. A two-factor solution was obtained with one factor defined as dental hygiene cognition and the second factor defined as dental hygiene clinical performance. Factor scores were subsequently used in a linear predictive model to assess the shared and unique contribution of factors to the one-shot clinical licensure examination score. The shared contribution of both factors only accounted for 13.9 percent of variance in the outcome measure of one-shot clinical licensure examination scores. The lack of concordance between previously validated measures of dental hygiene student competency or predictors of student success (Overall GPA, NBDHE, and Portfolios) and a one-shot clinical licensure examination (CRDTS) raises serious concern about the validity of our current dental hygiene licensing procedure which uses the CRDTS clinical examination to make decisions about granting licenses to practice. PMID- 15749948 TI - Increasing general dentists' provision of care to child patients through changes in the undergraduate pediatric dentistry program. AB - Reduced caries rates and an increased percentage of children with dental insurance have made it more difficult for dental schools to provide undergraduates with sufficient numbers of pediatric dental patients requiring restorative procedures. This may result in graduates who are not competent and are reluctant to treat children after graduation. To ensure the quality of the undergraduate clinical training program, the Division of Pediatric Dentistry at the University of Manitoba changed from a comprehensive-based clinic to a block system in 1998-99. Specific communities with limited access to dental care (neighboring core area schools and Hutterite colonies) were specifically targeted as potential sources for child patients. This format increased the exposure of students to patient management as well as to complex pediatric dentistry procedures. To assess the learning experiences before and after the changes to the clinical pediatric dentistry program, sixty general dentists who had graduated from the University of Manitoba were randomly selected using the Manitoba Dental Association Directory. Surveys were sent to twenty general dentists who graduated in each of the following years: 1993, 2000, and 2002. Forty-five dentists responded, fifteen from each of the three surveyed classes. Dentists who graduated after the changes to the program (2000, 2002) reported that they performed a greater number of complex pediatric dentistry procedures and treated more toddler and preschool children than the group that graduated before the changes (1993). Referrals to pediatric dentistry specialists were higher in the 1993 group than in the 2000 and 2002 groups. In conclusion, an adequate pool of pediatric patients is critical to provide dental students with sufficient learning experiences. The dentists who graduated from the program after the changes were implemented are providing more comprehensive treatment to younger children. PMID- 15749949 TI - Creating learner-centered classrooms: use of an audience response system in pediatric dentistry education. AB - Research suggests that the exclusive use of lecture in the classroom hinders student learning. The advent of compact electronic wireless audience response systems has allowed for increased student participation in the classroom. Such technology is utilized in medical education. This article describes the use of an audience response system in a "quiz bowl" format to facilitate and improve the comprehension of student dentists in core concepts in pulp therapy for the pediatric patient. PMID- 15749950 TI - Does school-based dental screening for children increase follow-up treatment at dental school clinics? AB - The purpose of this study was to evaluate the effectiveness of a screening program and referral system in stimulating dental attendance of children in need of treatment at Bapuji Dental College and Hospital of Davangere, India. A total of fourteen schools in the Davangere area were selected randomly and divided into two groups: seven schools that had a dental screening program (study group, n=2100 children), and seven schools that did not have one (control group, n=2400 children). The attendance rate by members of the study group was determined during the three-month period from the date of initiating the school screening program. During this same period the students who visited the college from the control group underwent a dental examination. Chi-square tests were used to test the difference between different variables. The response rate for seeking treatment was 31 percent for the study group (34.2 percent for males; 26.2 percent for females) and 10 percent for the control group (9.6 percent for males; 10.5 percent for females). In both the groups, the treatment need was highest for dental caries (study group=36.3 percent, control group=11.1 percent) and least for fluorosis (study group=21.2 percent, control group=1.2 percent). The study demonstrated that screening and motivation significantly improved the percentage of school children who sought dental care. PMID- 15749951 TI - Use of information and communication technology among dental students at the University of Jordan. AB - The aim of this study was to investigate the current knowledge, skills, and opinions of undergraduate dental students at the University of Jordan with respect to information communication technology (ICT). Dental students from the second, third, fourth, and fifth years were asked to complete a questionnaire presented in a lecture at the end of the second semester in the 2002-03 academic year. The response rate was 81 percent. Besides free and unlimited access to computers at the school of dentistry, 74 percent of the students had access to computers at home. However, 44 percent did not use a computer regularly. Male students were more regular and longer users of computers than females (p<0.001). A significant number of students (70 percent) judged themselves competent in information technology (IT) skills. More males felt competent in basic IT skills than did females (p<0.05). More than two-thirds acquired their computer skills through sources other than at the university. The main educational use of computers was accessing the Internet, word processing, multimedia, presentations, Medline search, and data management. More clinical students felt competent in word-processing skills (p<0.05) and many more used word processing for their studies (p<0.001) than did preclinical students. More males used word processing for their studies than females (p<0.001). Students used computers for personal activities more frequently than for academic reasons. More males used computers for both academic (p<0.01) and personal activities (p<0.001) than did females. All students had access to the Internet at the university, and 54 percent had access at home. A high percentage of students (94 percent) indicated they were comfortable using the Internet, 75 percent said they were confident in the accuracy, and 80 percent said they were confident in the relevance of information obtained from the Internet. Most students (90 percent) used email. Most students (83 percent) supported the idea of placing lectures on the web, and 61.2 percent indicated that this would not influence lecture attendance. Students used the Internet more for personal reasons than for the study of dentistry. More clinical students used the Internet for dentistry than preclinical students (p<0.001). More males than females used the Internet for dentistry (p<0.01) as well as for pleasure (p<0.01). Time and availability were the main obstacles to Internet use. Dental students at the University of Jordan have access to substantial IT resources and demonstrated attitudes toward the computer and Internet technology and use that were similar to other students in other nations. However, the educational use of ICT among Jordanian students remains low. PMID- 15749952 TI - Molecular physiology and pathology of Ca2+-conducting channels in the plasma membrane of mammalian sperm. AB - Current evidence indicates that mechanisms controlling the intracellular Ca2+ concentration play pivotal roles in determining sperm fertilizing ability. Multiple Ca2+-permeable channels have been identified and characterized in the plasma membrane and in the acrosome membrane of mammalian sperm. This review summarizes the recent findings and assesses the evidence suggesting that these channels play roles in controlling a host of sperm functions ranging from motility to the acrosome reaction, and describes recent advances in the identification of the underlying gene defects of inherited sperm Ca2+ channelopathies. PMID- 15749953 TI - Role of actin cytoskeleton in mammalian sperm capacitation and the acrosome reaction. AB - In order to fertilize, the mammalian spermatozoa should reside in the female reproductive tract for several hours, during which they undergo a series of biochemical modifications collectively called capacitation. Only capacitated sperm can undergo the acrosome reaction after binding to the egg zona pellucida, a process which enables sperm to penetrate into the egg and fertilize it. Polymerization of globular (G)-actin to filamentous (F)-actin occurs during capacitation, depending on protein kinase A activation, protein tyrosine phosphorylation, and phospholipase D activation. F-actin formation is important for the translocation of phospholipase C from the cytosol to the sperm plasma membrane during capacitation. Prior to the occurrence of the acrosome reaction, the F-actin should undergo depolymerization, a necessary process which enables the outer acrosomal membrane and the overlying plasma membrane to come into close proximity and fuse. The binding of the capacitated sperm to the zona pellucida induces a fast increase in sperm intracellular calcium, activation of actin severing proteins which break down the actin fibers, and allows the acrosome reaction to take place. PMID- 15749954 TI - Expression and proteasomal degradation of the major vault protein (MVP) in mammalian oocytes and zygotes. AB - Major vault protein (MVP), also called lung resistance-related protein is a ribonucleoprotein comprising a major part (>70%) of the vault particle. The function of vault particle is not known, although it appears to be involved in multi-drug resistance and cellular signaling. Here we show that MVP is expressed in mammalian, porcine, and human ova and in the porcine preimplantation embryo. MVP was identified by matrix-assisted laser-desorption ionization-time-of-flight (MALDI-TOF) peptide sequencing and Western blotting as a protein accumulating in porcine zygotes cultured in the presence of specific proteasomal inhibitor MG132. MVP also accumulated in poor-quality human oocytes donated by infertile couples and porcine embryos that failed to develop normally after in vitro fertilization or somatic cell nuclear transfer. Normal porcine oocytes and embryos at various stages of preimplantation development showed mostly cytoplasmic labeling, with increased accumulation of vault particles around large cytoplasmic lipid inclusions and membrane vesicles. Occasionally, MVP was associated with the nuclear envelope and nucleolus precursor bodies. Nucleotide sequences with a high degree of homology to human MVP gene sequence were identified in porcine oocyte and endometrial cell cDNA libraries. We interpret these data as the evidence for the expression and ubiquitin-proteasome-dependent turnover of MVP in the mammalian ovum. Similar to carcinoma cells, MVP could fulfill a cell-protecting function during early embryonic development. PMID- 15749956 TI - Serine protease activity, bovine sperm protease, 66 kDa (BSp66), is present in hamster sperm and is involved in sperm-zona interaction. AB - Bovine sperm protease, 66 kDa (BSp66) is a serine protease previously characterized in bovine spermatozoa. Like other proteases, it may be present in sperm from other mammalian species different from bovine, playing a role in the fertilization process. In this study, we looked for BSp66 in hamster spermatozoa using heterologous antibodies against bovine BSp66. An immunoreactive protein was detected by Western blotting in mature and immature sperm. The detected protein had two isoforms similar to the ones reported in bovine sperm. Furthermore, indirect immune detection by fluorescence and electron microscopy assays, showed BSp66 signal at the acrosomal region similar to bovine sperm. As it was determined in bovine sperm, the acrosomal reaction displays the antigen within the acrosomal content. When live hamster sperm was incubated with polyclonal antibody against bovine BSp66 a decrease in the number of sperm bound to zona pellucida in homologous IVF and an impairment of head-head agglutination, were observed. These results suggest that a protease homologous to bovine BSp66 is present in golden hamster spermatozoa, with a conserved molecular mass and cellular location. Moreover, hamster BSp66 is probably involved in zona pellucida recognition. PMID- 15749955 TI - Inhibition of phosphatidylinositol 3-kinase modifies boar sperm motion parameters. AB - Motility is the most widely used indicator of sperm quality. Besides modulation by the cAMP pathway little is known regarding the intracellular pathways that regulate boar sperm motility. Recently the role of phosphatidylinositol 3-kinase (PI3-K) in the regulation of human sperm motility has been described. Therefore, the aim of this study was to investigate the role of PI3-K in boar sperm kinematics by using the specific PI3-K inhibitor, LY294002. Boar sperm was incubated up to 1 h in non-capacitating medium in the presence or absence of the cAMP analog, 8Br-cAMP or the PI3-K inhibitor, LY294002 or both. Boar sperm incubated in capacitating medium was treated in the presence or absence of LY294002. First, we have clearly identified that PI3-K is present in whole lysates of boar spermatozoa. Inhibition of PI3-K significantly increased boar sperm straight-line velocity, circular velocity and average velocity without an effect on the percentage of progressively motile spermatozoa in both media. Inhibition of PI3-K induced the same effects on boar sperm velocities as activation of the cAMP/protein kinase A (PKA) pathway and treatment with the PI3 K inhibitor, LY294002 had neither summatory nor synergic effects on boar sperm motion parameters when treated simultaneously with the cAMP analog 8Br-cAMP. Our data suggest that PI3-K plays a negative role, regulating boar sperm motion parameters through a possible inhibition of the cAMP/PKA activating pathway, and since some Computer Aided Sperm Analysis (CASA)-derived parameters have been related to field fertility our results point to the possibility of modulating sperm motile quality by modifying the PI3-K cellular pathway. PMID- 15749957 TI - Short-term nutritional supplementation of ewes in low body condition affects follicle development due to an increase in glucose and metabolic hormones. AB - This study tested whether the effects of a short period of nutritional supplementation given to ewes during the luteal phase on follicle development and ovulation rate is associated with an increase in circulating concentrations of FSH, glucose or metabolic hormones. Oestrus was synchronised with two prostaglandin injections given 9 days apart and the supplement consisted of corn grain and soybean meal. Corriedale ewes with low body condition were randomly assigned to 2 groups: the control group (C; n = 10) received a maintenance diet while the short-term supplemented group (STS; n = 10) received double the maintenance diet over days 9 to 14 of the oestrous cycle (day 0 = ovulation). Ovaries were examined daily by ultrasound and blood was sampled three times a day during the inter-ovulatory interval for measuring reproductive and metabolic hormones. On days 9, 11 and 14 of the oestrous cycle, half of the ewes from each group (n = 5) were bled intensively to determine the concentrations of glucose, insulin, IGF-I and leptin. Plasma FSH, progesterone, oestradiol and androstenedione concentrations were similar among groups. Dietary supplementation increased plasma insulin concentrations from the first to the sixth day of supplementation and increased glucose concentrations on the third day, compared with control ewes. Plasma leptin concentrations were higher in STS ewes from the second to the fifth day of supplementation. The pattern of IGF-I concentrations was similar among groups. In STS ewes, the nutritional treatment prolonged the lifespan of the last non-ovulatory follicle, so fewer follicular waves developed during the cycle. In STS ewes, increased concentrations of glucose, insulin and leptin one day before ovulatory wave emergence were associated with increased numbers of follicles growing from 2 to 3 mm and with stimulation of the dominant follicle to grow for a longer period. We suggest that the mechanism by which short-term nutritional supplementation affects follicle development does not involve an increase in FSH concentrations, but may involve responses to increased concentrations of glucose, insulin and leptin, acting directly at the ovarian level. This effect is acute, since concentrations of all three substances decrease after reaching peak values on the third day of supplementation. The status of follicle development at the time of maximum concentrations of glucose and metabolic hormones may be one of the factors that determines whether ovulation rate increases or not. PMID- 15749958 TI - Cyclic changes of the ovarian surface epithelium in the rat. AB - The ovarian surface epithelium (OSE) plays pivotal roles during ovulation and postovulatory wound repair. In this paper we describe the proliferative activity of the OSE through the estrous cycle in adult cycling rats, by immunohistochemical detection of DNA-incorporated bromodeoxyuridine (BrdU). Immunohistochemical detection of estrogen receptor alpha (ERalpha) and progesterone receptor was also performed. The cycle of the OSE consists of a proliferative phase (that lasts for two consecutive estrous cycles) and a quiescent phase of variable duration. Cyclic changes in the OSE were related to the underlying ovarian structure. OSE areas covering growing follicles entered into the proliferative phase during the transition from proestrus to estrus, with the appearance of fast-growing class 1 follicles, destined to ovulate at the end of the current estrous cycle. A labeling index (after pulse-labeling BrdU treatment) of about 7% was maintained throughout the estrous cycle in parallel to follicle growth. Cumulative BrdU-labeling (after daily BrdU treatment) indicated that about 1/3 of the total OSE cell proliferation was related to follicle growth. Following ovulation, OSE cells covering newly-formed corpora lutea showed a labeling index of about 50% that decreased through metestrus and diestrus (about 13% and 3%, respectively), returning to basal levels by proestrus. Cumulative BrdU-labeling indicated that about 2/3 of the total proliferative activity was related to ovulation repair/luteinization. The remaining OSE covering ovarian stroma or structurally regressing corpora lutea of previous cycles showed negligible BrdU labeling. The equivalent proliferative activity found in the OSE covering newly-formed corpora lutea in indomethacin-treated rats lacking rupture of the OSE at the apex, demonstrated that ovulation-triggered proliferation was not dependent on the loss of integrity of the OSE at the ovulation site. OSE cells expressed ERalpha throughout the cycle, but no differential expression was found between proliferating and quiescent OSE areas. On the contrary, OSE cells did not express PR at any time of the cycle. These data indicate the existence of a cycle of the OSE, related to the cyclic changes in the underlying ovarian structure and strongly suggest that the proliferative activity of the OSE is regulated by local microenvironmental rather than by systemic factors. PMID- 15749959 TI - Endometrial tumor necrosis factor alpha (TNFalpha) is a likely mediator of early luteal phase mifepristone-mediated negative effector action on the preimplantation embryo. AB - Cytokines and growth factors are important mediators of progesterone-regulated endometrial receptivity and embryo development. Early luteal phase administration of a potent antiprogestin-like mifepristone to the rhesus monkey results in endometrial desynchrony, loss of embryo viability and implantation failure. In the present study, administration of mifepristone (2 mg/kg body weight, s.c.) on day 2 after ovulation resulted in a significant increase (P < 0.01) in the level of tumor necrosis factor alpha (TNFalpha) in glandular and vascular compartments of endometrium, and in endometrial secretion and luminal fluid on day 6 after ovulation in the rhesus monkey. There was an associated lag in embryonic development, characterized by delayed mitochondrial maturity, poorly developed junctional complexes, a relative absence of intra-cytoplasmic filaments and a high degree of intra-cellular degenerative features. Exposure of TNFalpha (0, 0.5, 5, 50 ng/ml) to preimplantation stage mouse embryos in vitro showed a dose dependent arrest in growth and development at both morula and blastocyst stages along with ultra-structural features of degeneration similar to those observed in embryos collected from early luteal phase mifepristone-treated monkeys. The de novo synthesized and released proteins in terms of trichloroacetic acid precipitable 35S by morulae and blastocysts in vitro showed a marked depression following exposure to TNFalpha compared with control embryos. Based on the above observation and the fact that preimplantation stage embryos express receptors for TNFalpha, we suggest that increased levels of TNFalpha in endometrial and luminal compartments around the time of uterine receptivity following early luteal phase administration of mifepristone adversely affect the growth and viability of preimplantation stage embryos. PMID- 15749961 TI - Immunohistochemical studies on the progesterone receptor (PR) in the sow uterus during the oestrous cycle and in inseminated sows at oestrus and early pregnancy. AB - Physiological changes in the sow uterus involve the regulation by progesterone and its receptor proteins (PR). Therefore, the aim of the present study was to investigate the localization of PR during different stages of the oestrous cycle and in inseminated sows during early pregnancy by use of immunohistochemistry. Uterine samples were collected from cyclic and inseminated sows at different stages of the oestrous cycle and early pregnancy. The samples were fixed in 10% formaldehyde and embedded in paraffin. Immunohistochemistry was done by use of a mouse monoclonal antibody to PR. The highest PR immunostaining in the surface epithelium was observed at oestrus/5-6 h after artificial insemination (AI) and early dioestrus/70 h after AI. In the glandular epithelium, the highest level of PR was found at oestrus with the lowest at late dioestrus/d 19. Higher levels of PR were observed in inseminated groups compared with cyclic sows. In the myometrium, a high level of PR was found at oestrus, while stromal PR cells were constantly present throughout the oestrous cycle and at different stages of early pregnancy. In conclusion, this study shows that the immunopresence of PR in the sow uterus differed between uterine compartments at the same reproductive stage. Differences were also found for some uterine compartments between cyclic and inseminated/early pregnant sows. The relatively consistent immunostaining of PR in the stroma strengthens a stromal role in the regulation of physiological activities in the sow uterus during the oestrous cycle as well as early pregnancy. PMID- 15749962 TI - Long-term effects of deslorelin implants on reproduction in the female tammar wallaby (Macropus eugenii). AB - The contraceptive and endocrine effects of long-term treatment with implants containing the GnRH agonist deslorelin were investigated in female tammar wallabies (Macropus eugenii). Fertility was successfully inhibited for 515 +/- 87 days after treatment with a 5 mg deslorelin implant (n = 7), while control animals gave birth to their first young 159 +/- 47 days after placebo implant administration (n = 8). The duration of contraception was highly variable, ranging from 344 to 761 days. The strict reproductive seasonality in the tammar wallaby was maintained once the implant had expired. This inhibition of reproduction was associated with a significant reduction in basal LH concentrations and a cessation of oestrous cycles, as evidenced by low progesterone concentrations. There was evidence to suggest that some aspect of either blastocyst survival, luteal reactivation, pregnancy or birth may be affected by deslorelin treatment in some animals. These results show that long term inhibition of fertility in the female tammar wallaby is possible using slow release deslorelin implants. The effects of deslorelin treatment were fully reversible and there was no evidence of negative side effects. Slow-release GnRH agonist implants may represent a practicable method for reproductive management of captive and semi-wild populations of marsupials. PMID- 15749960 TI - The IGF system in the neonatal ovine uterus. AB - Postnatal development of the ovine uterus primarily involves uterine gland morphogenesis or adenogenesis. Adenogenesis involves the budding differentiation of the glandular epithelium (GE) from the luminal epithelium (LE) and then GE proliferation and coiling/branching morphogenetic development within the stroma between birth (postnatal day or PND 0) and PND 56. Insulin-like growth factor (IGF)-I and IGF-II mRNAs were previously found to be expressed only in the endometrial stroma, whereas the IGF receptor (IGF-1R) mRNA was most abundant in epithelia and in stroma, suggesting that an intrinsic IGF system regulates postnatal development of the uterus. Given that the biological activities of IGFs are modulated by a family of six IGF binding proteins (IGFBPs) and specific proteases, the objective was to determine the effects of age and estrogen disruption on expression of IGFs, IGFBPs and pregnancy-associated plasma protein A (PAPP-A or IGFBP-4 protease) in the ovine uterus. In Study One, circulating levels of IGF-I and IGF-II in the serum of neonatal ewes did not change between PND 0 and PND 56. Levels of immunoreactive IGF-I, IGF-II and IGF-1R protein were most abundant on the apical surface of the endometrial LE and GE. RT-PCR analyses detected expression of IGFBPs (3, 4, 5 and 6) as well as PAPP-A mRNAs in the uterus, but not IGFBP-1 and IGFBP-2 mRNAs. IGFBP-3 and IGFBP-4 mRNAs were expressed specifically in the endometrial stroma and myometrium and increased after birth. PAPP-A mRNA was expressed specifically in the endometrial stroma and increased after birth. In Study Two, ewes were treated from birth with estradiol 17beta valerate (EV), which reduces uterine growth and inhibits endometrial adenogenesis. On PNDs 14 and 56, IGFBP-3 mRNA was decreased in the uterus of EV treated ewes, but IGF-1R and IGFBP-4 mRNAs were not affected. PAPP-A mRNA was increased by EV treatment on PND 14, but decreased on PND 56. These results support the hypothesis that an intrinsic IGF system in the uterus regulates epithelial-stromal interactions important for postnatal uterine growth and endometrial gland morphogenesis in the sheep. PMID- 15749963 TI - Membrane fluidity and the ability of domestic bird spermatozoa to survive cryopreservation. AB - The ability to survive cryopreservation varies in spermatozoa from different bird species. Among the biological factors potentially responsible for such differences, species variations in membrane fluidity have a role in the restoration of the physiological state after freezing. Membrane fluidity may be assessed by measuring fluorescence polarization anisotropy with a fluorescent dye. Anistropy values are proportional to membrane rigidity and consequently inversely proportional to membrane fluidity. In the present study, polarization anisotropy of spermatozoa originating from species differing in the freezability of their semen (chicken, turkey and guinea fowl) was measured in addition to lipid composition (cholesterol/phospholipid ratio), sperm viability (membrane permeability to eosine) and morphological integrity before and after cryopreservation. The percentages of viable and normal spermatozoa in fresh sperm were highest in the chicken (87%), lowest in guinea fowl (64%), and intermediate in turkeys (69%). Anisotropy values were highest in guinea fowl (0.205), lowest in chickens (0.155), and intermediate in turkeys (0.180). As a consequence, membrane fluidity was highest in chickens and lowest in guinea fowl. Cryopreservation significantly decreased sperm viability and morphological integrity and increased anisotropy in all species but did not change the inter species hierarchy. Initial cholesterol/phospholipid ratios were lower in chickens than in guinea fowl, and intermediate in turkeys (0.25, 0.26 and 0.29, respectively). Cryopreservation induced a severe decrease in cholesterol/phospholipid ratios in turkeys and guinea fowl. Sperm membrane fluidity in chickens, turkeys and guinea fowl behaves as an indicator of sperm freezability in these species. Inter species differences for this parameter may be partly explained by differences in initial cholesterol/phospholipids content of spermatozoa. On the other hand, the rigidifying process induced by cryopreservation is not related to lipid damage by the same mechanisms. PMID- 15749965 TI - Predictors of violent behavior in an early adolescent cohort: similarities and differences across genders. AB - The authors assessed a cohort of 2,335 students from the Minneapolis, Minnesota, area to identify predictors of violent behavior and to determine whether the predictors varied by gender. The sample was 76% White; boys and girls were equally represented. The majority lived with two parents. A measure of violent behavior collected at the end of the eighth-grade year (2000) was entered into Poisson regression against baseline data collected at the beginning of the seventh-grade year (1998). Predictors of violent behavior influencing both boys and girls included depressive symptoms, perceived invulnerability to negative future events, paternal nonauthoritative behavior, and drinking alcohol. Additional predictors of violent behavior specific to girls included both risk and protective factors. PMID- 15749966 TI - Development of measures of organizational leadership for health promotion. AB - This study describes the development and psychometric evaluation of four scales measuring leadership for health promotion at an organizational level in the baseline survey (n=144) of the Alberta Heart Health Project. Content validity was established through a series of focus groups and expert opinion appraisals, pilot testing of a draft based on capacity assessment instruments developed by other provinces involved in the Canadian Heart Health Initiative, and the literature. Psychometric analyses provided empirical evidence of the construct validity and reliability of the organizational leadership scales. Principal component analysis verified the unidimensionality of the leadership scales of (a) Practices for Organizational Learning, (b) Wellness Planning, (c) Workplace Climate, and (d) Organization Member Development. Scale alpha coefficients ranged between .79 and .91 thus establishing good to high scale internal consistencies. These measures can be used by both researchers and practitioners for the assessment of organizational leadership for health promotion and heart health promotion. PMID- 15749967 TI - Intention or experience? Predictors of continued breastfeeding. AB - Despite the known benefits of breastfeeding, many women do not breastfeed their infants or stop breastfeeding early. This study examines the effects of prenatal intention and initial breastfeeding experiences on breast-feeding initiation and duration among 1,665 U.S. women completing questionnaires on infant feeding practices. Outcomes included no initiation of breastfeeding at birth and termination at <10 weeks, 10 to <20 weeks, or 20 to <30 weeks. Predictor variables included intended breast feeding duration and early breast feeding experiences with analyses controlling for demographic characteristics, previous breastfeeding experience, and prenatal intentions to work after delivery. Prenatal intentions to never initiate or to stop breastfeeding early were significant risk factors for all breastfeeding outcomes. Initial breastfeeding experiences were significant risk factors for early termination. This study supports using the intention construct from the theory of reasoned action to predict initiation of behavior but suggests the need to include initial experience when predicting maintenance of behavior. PMID- 15749968 TI - Impact analysis and mediation of outcomes: the Going Places program. AB - The purpose of the study was to evaluate the impact of the Going Places Program and mediation of treatment effects. Seven middle schools were randomized to intervention or comparison conditions and students (n=1,320) in two successive cohorts provided five waves of data from sixth through eighth grade. The Going Places Program included classroom curriculum, parent education, and school environment components. Latent growth curve analyses demonstrated significant treatment group effects on outcome expectancies, friends who smoke, and smoking. Friends who smoke mediated the program effect on adolescents' smoking progression. The protective effect of the Going Places Program on smoking progression was due in part to the prevention of increases in friends who smoke. PMID- 15749969 TI - Organ donor registration preferences among Hispanic populations: which modes of registration have the greatest promise? AB - There is little debate over Hispanics' need for viable organs. Although organ donor registries can potentially assist in alleviating this need, the U.S. Department of Health and Human Services (DHHS) has called on researchers to investigate methods to maximize such potential. This research effort answers the aforementioned call by surveying Hispanics, both those who prefer to use Spanish (SP, n=239) and those who prefer not to use Spanish (NSP, n=364) about the methods of donor registration that they would most likely utilize. When provided with different options for organ donor registration, signing up by applying for a driver's license, through a place of worship, or through a doctor's office were among the most popular methods; over the telephone, tax forms, or through the use of home computer were among the least popular. Furthermore, the DHHS-suggested idea of registering via organ donor kiosks was very well received. PMID- 15749970 TI - Healthy eating and exercise: strategies for weight management in the rural midwest. AB - Obesity prevalence has increased dramatically in the United States. Rural areas have been especially affected, yet few weight management studies have been conducted in these populations. This study was designed to assess weight management attitudes and strategies used when rural adults in particular attempt to lose weight, employing measures that are more specific than those reported in similar studies. The survey was completed by 123 adults aged 19 to 71, with a 29% response rate. Scales demonstrated good to excellent internal consistency, and test-retest reliability. Persons currently trying to lose weight were more likely to report self-monitoring behaviors and goal setting but no more likely to report use of specific behavioral strategies compared to persons not trying to lose weight. The results suggest considerable interest in weight management among rural adults and provide a starting point for planning future weight management interventions in this and similar populations. PMID- 15749971 TI - Modeling psychological empowerment among youth involved in local tobacco control efforts. AB - The American Legacy Foundation funded 13 state health departments for their Statewide Youth Movement Against Tobacco Use in September 2000. Its goal was to create statewide tobacco control initiatives implemented with youth leadership. The underlying theory behind these initiatives was that tobacco control efforts can best be accomplished by empowering youth. To evaluate these initiatives, the authors developed a conceptual framework for youth empowerment that was used as a guide in developing standardized cross-site measures. This article describes the domains and attributes used to operationalize psychological empowerment as an outcome of youth involvement in these initiatives and presents results of our two stage structural equation modeling. We conclude with a summary of lessons learned to date and recommendations for applying these findings to work in the field. PMID- 15749972 TI - Biliary atresia: feasibility of mangafodipir trisodium-enhanced MR cholangiography for evaluation. AB - The study was approved by the institutional review board, and informed consent was obtained from the patients' parents. Twenty-three consecutive infants suspected of having biliary atresia (BA) were prospectively examined by using mangafodipir trisodium (Mn-DPDP)-enhanced magnetic resonance (MR) cholangiography. Sequential T1-weighted spoiled gradient-echo MR cholangiograms were obtained 1, 2, and 3 hours after intravenous administration of Mn-DPDP. The possibility of BA was excluded if bowel excretion of contrast material was noted at contrast material-enhanced MR cholangiography. The diagnostic specificity and accuracy of contrast-enhanced MR cholangiography were compared with those of conventional MR cholangiography, technetium 99m Tc ((99m)Tc)-disofenin (DISIDA) scintigraphy, and the triangular cord sign at ultrasonography (US). MR cholangiography was used to accurately distinguish four cases of BA from 19 cases of other cholestatic liver diseases, without false-positive results. Conventional MR cholangiography, (99m)Tc-DISIDA scintigraphy, and the triangular cord sign at US respectively yielded false-positive results of 42% (eight of 19 infants), 35% (six of 17 infants), and 11% (two of 19 infants) in patients without BA. Mn-DPDP enhanced MR cholangiography appears to be a promising modality for early diagnosis of BA as the cause of neonatal cholestasis. PMID- 15749973 TI - Blunt splenic injury: usefulness of transcatheter arterial embolization in patients with a transient response to fluid resuscitation. AB - PURPOSE: To evaluate the use of transcatheter arterial embolization (TAE) in hemodynamically unstable patients with blunt splenic injury in whom there is a transient response to initial fluid resuscitation. MATERIALS AND METHODS: Human subject committee approval and informed consent were obtained. Angiography was performed in patients with contrast material extravasation and/or splenic injury of grade III or higher (American Association for the Surgery of Trauma criteria) at computed tomography (CT). TAE was performed when angiograms showed disruption of terminal splenic branches or arterial extravasation. Among 104 patients with splenic injury, the 15 patients (10 male, five female; mean age, 36.2 years) with a transient response to fluid resuscitation were the subjects of this study. A post hoc analysis was performed for CT grades, angiographic findings, associated injuries, and hemodynamic status in the subjects. RESULTS: Among 15 patients with a transient response, two had grade III, 11 had grade IV, and two had grade V injuries at CT. Six patients had associated injuries that required TAE. TAE of the spleen and associated injuries was successfully performed in all patients. The mean systolic blood pressure and shock index at the start of TAE were 84.2 mm Hg +/- 9.2 (standard deviation) and 1.46 +/- 0.30, respectively, and those at the completion of TAE were 132.1 mm Hg +/- 18.7 and 0.77 +/- 0.21, respectively (P < .001). The fluid infusion rate within 24 hours after the completion of TAE (132.1 mL/h +/- 71.1) was lower than that from the completion of the initial fluid resuscitation until the completion of TAE (1230.6 mL/h +/- 264.8) (P < .001). CONCLUSION: TAE for blunt splenic injury can be performed successfully even in hemodynamically unstable patients with a transient response to initial fluid resuscitation. PMID- 15749974 TI - Leg perforator vein incompetence: functional anatomy. AB - PURPOSE: To retrospectively determine the anatomic patterns of reflux of incompetent perforator veins (IPVs) at the sites of their highest prevalence in relation to the anatomic distribution of valvular incompetence in the veins of the calf and thigh, with emphasis on the deep system, across the clinical spectrum of chronic venous disease (CVD). MATERIALS AND METHODS: This study was granted institutional ethics committee approval; the need for patient consent was waived. Five hundred five limbs in 359 consecutive subjects who were suspected of having CVD but did not have arterial disease, prior venous thrombosis (<1 year), venous or orthopedic surgery, or vascular malformations were clinically stratified for CVD according to the clinical, etiologic, anatomic, and pathophysiologic (CEAP) system and underwent venous hemodynamic investigation with duplex ultrasonography. One hundred thirty limbs were CEAP clinical classes C(0-1), 262 limbs were classes C(2-3), and 113 limbs were classes C(4-6). IPV reflux patterns and anatomic distribution of deep venous reflux in the lower limb were determined across the clinical classes of CVD. Statistical analysis was performed with Spearman rank correlation, chi(2), and Mann-Whitney testing. RESULTS: Valvular incompetence in limbs with IPVs increased with CEAP clinical class (P < .01) in femoral, popliteal, posterior tibial, peroneal, gastrocnemial, and soleal veins; reflux was distributed evenly across these veins. Of 554 IPVs found, 377 (68.0%) occurred at four sites: middle third of medial calf (n = 165 [29.8%]), lower third of medial calf (n = 85 [15.3%]), middle third of medial thigh (n = 73 [13.2%]), and middle third of posterior calf (n = 54 [9.7%]). IPVs with superficial and deep reflux in adjoining veins, as compared with IPVs with superficial reflux alone, increased as clinical class increased from C(2) to C(6) (P < .02) at all four sites of highest IPV prevalence; determined in detail, reflux patterns of IPVs were linked to CEAP clinical class (P < .05) but not anatomic site (P > .2). Most IPVs in C(1-3) limbs had superficial reflux alone. IPVs with superficial reflux outnumbered IPVs with superficial and deep reflux even in C(4-6) limbs, where deep venous incompetence was most prevalent. Axial venous reflux (proximal-to-distal) changes (P > .4) were small in superficial and deep veins across the spectrum of CEAP clinical classes C(2-6). CONCLUSION: Patterns of perforator reflux were linked to clinical severity of CVD in the CEAP classification and displayed an even distribution anatomically. IPVs with deep and superficial reflux in adjoining veins increased with CEAP clinical class, in line with valvular incompetence in the deep veins of the calf and thigh. PMID- 15749975 TI - Distribution of cerebral blood flow in the circle of Willis. AB - PURPOSE: To prospectively determine the effect of anatomic variations in the circle of Willis on volume flow in the internal carotid arteries (ICAs) and basilar artery (BA). MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. Phase-contrast magnetic resonance (MR) angiography was used to measure the volume flow in the BA and ICAs in 208 patients (182 men, 26 women; mean age, 60 years) with symptomatic atherosclerosis or risk factors for atherosclerosis. Patients with steno-occlusive disease were excluded, and flow values were normalized for age. Three-dimensional time-of flight MR angiograms were used to assess the anatomy of the circle of Willis. Differences in volume flow between a complete circle of Willis, a circle with a missing A1 segment, and a circle with a fetal-type posterior cerebral artery were analyzed (analysis of variance and Scheffe post hoc tests). RESULTS: The ICA volume flow in subjects with a complete configuration of the circle of Willis was 245 mL/min +/- 65 (standard deviation). Flow in the contralateral ICA was significantly increased (P < .01) in subjects with a missing A1 segment (303 mL/min +/- 56) compared with control subjects and compared with flow on the ipsilateral side (214 mL/min +/- 94; P < .01). In subjects with a unilateral or bilateral fetal-type posterior cerebral artery, the ICA volume flow was increased (P < .01) and the BA volume flow was decreased (P < .01) in comparison with the flow in subjects with no fetal-type circle of Willis. CONCLUSION: Large asymmetries in volume flow between the right and left ICAs or decreased volume flow in the BA is not necessarily caused by vascular disease but may be caused by variations in the anatomy of the circle of Willis. PMID- 15749976 TI - Cell tagging with clinically approved iron oxides: feasibility and effect of lipofection, particle size, and surface coating on labeling efficiency. AB - PURPOSE: To evaluate the effect of lipofection, particle size, and surface coating on labeling efficiency of mammalian cells with superparamagnetic iron oxides (SPIOs). MATERIALS AND METHODS: Institutional Review Board approval was not required. Different human cell lines (lung and breast cancer, fibrosarcoma, leukocytes) were tagged by using carboxydextran-coated SPIOs of various hydrodynamic diameters (17-65 nm) and a dextran-coated iron oxide (150 nm). Cells were incubated with increasing concentrations of iron (0.01-1.00 mg of iron [Fe] per milliliter), including or excluding a transfection medium (TM). Cellular iron uptake was analyzed qualitatively at light and electron microscopy and was quantified at atomic emission spectroscopy. Cell visibility was assessed with gradient- and spin-echo magnetic resonance (MR) imaging. Effects of iron concentration in the medium and of lipofection on cellular SPIO uptake were analyzed with analysis of variance and two-tailed Student t test, respectively. RESULTS: Iron oxide uptake increased in a dose-dependent manner with higher iron concentrations in the medium. The TM significantly increased the iron load of cells (up to 2.6-fold, P < .05). For carboxydextran-coated SPIOs, larger particle size resulted in improved cellular uptake (65 nm, 4.37 microg +/- 0.08 Fe per 100 000 cells; 17 nm, 2.14 microg +/- 0.06 Fe per 100 000 cells; P < .05). Despite larger particle size, dextran-coated iron oxides did not differ from large carboxydextran-coated particles (150 nm, 3.81 microg +/- 0.46 Fe per 100 000 cells; 65 nm, 4.37 microg +/- 0.08 Fe per 100 000 cells; P > .05). As few as 10 000 cells could be detected with clinically available MR techniques by using this approach. CONCLUSION: Lipofection-based cell tagging is a simple method for efficient cell labeling with clinically approved iron oxide-based contrast agents. Large particle size and carboxydextran coating are preferable for cell tagging with endocytosis- and lipofection-based methods. PMID- 15749977 TI - Angiosarcoma of the spleen: imaging characteristics in 12 patients. AB - PURPOSE: To retrospectively review clinical, pathologic, and imaging features of angiosarcoma of the spleen in 12 patients. MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was not required. Records of 12 cases of proved angiosarcoma of the spleen were accessed from the files of the Armed Forces Institute of Pathology. Clinical, pathologic, and imaging findings were reviewed. Presenting signs and symptoms were recorded, and the pathologist confirmed the diagnosis and determined gross and microscopic morphologic findings in each spleen. Radiologists reviewed available images to determine the size of the spleen and mass, amount of splenic involvement by the tumor (if enhanced at computed tomography [CT] and magnetic resonance [MR] imaging), and amount of nonviable tumor determined as decreased echogenicity at ultrasonography (US) and lack of enhancement at CT and MR imaging. Imaging and pathologic findings were compared. Five US, 10 CT, three MR, and two angiographic images were reviewed by two experienced abdominal radiologists. RESULTS: There were seven men and five women (age range, 36-86 years; mean, 55 years). The most common symptom was upper abdominal pain in eight (67%) patients: Pain was acute for 24 hours prior to admission in one patient and chronic (range, 1-6 months) in seven patients. At imaging, the spleen was enlarged (>12 cm in length) in nine patients. The most common finding, seen in seven (58%) patients, was a complex mass or masses in an enlarged spleen. Four of these patients had evidence of metastases and one had intraperitoneal hemorrhage. Two patients had solitary hypervascular tumors and liver metastases. One patient had a normal-sized spleen with multiple lesions that ranged 2-3 cm in size, as well as metastases to the spine. The 11th patient had two small lesions, with small calcifications in the periphery of one lesion. The 12th patient had intraabdominal hemorrhage around the spleen and no obvious mass at CT. Tumor necrosis was confirmed at histologic evaluation in nine patients. CONCLUSION: The most common clinical finding was upper abdominal pain. Angiosarcoma of the spleen could be suggested in the majority of cases (83%) by using the imaging features of splenic mass with evidence of metastatic disease. PMID- 15749978 TI - Differentiation of nonperforated from perforated appendicitis: accuracy of CT diagnosis and relationship of CT findings to length of hospital stay. AB - PURPOSE: To determine retrospectively the sensitivity and specificity of computed tomographic (CT) signs in differentiating acute nonperforated appendicitis from perforated appendicitis and to compare CT findings with the length of hospital stay. MATERIALS AND METHODS: Institutional Review Board approval was obtained for this study, and patient informed consent was obtained for record review for research purposes. Two radiologists were blinded to patient identification but were informed that all patients presented to the emergency department with abdominal pain and underwent appendectomy. Radiologists independently reviewed CT images of 86 consecutive patients (45 males, 41 females; mean age, 33.7 years; age range, 8.2-87.1 years) who presented to the emergency department with acute abdominal pain, who underwent CT after initial emergency department assessment, and who underwent appendectomy within the subsequent 24 hours. Individual findings and confidence level for the diagnosis of perforated appendicitis were noted. Consensus interpretation was performed with a third radiologist. The consensus CT findings were correlated with the surgical and pathologic findings by using chi(2) or Fisher exact tests for univariate analysis and logistic regression for multiple variable analysis. Wilcoxon rank sum tests were used to assess the association between consensus CT findings and length of hospital stay. RESULTS: Twenty-one (24%) of the 86 patients had appendiceal perforation. Extraluminal air and either moderate or severe periappendiceal inflammatory stranding were statistically significant independent predictors for perforation (P < .001). A focal defect in enhancing appendiceal wall was significantly associated with perforation (P < .001) and had a sensitivity of 58.8% and specificity of 85.7% on consensus review, with eight false-positive results. There was a strong association between the degree of periappendiceal inflammatory stranding and the length of hospital stay (P < .001). CONCLUSION: Extraluminal air and moderate or severe periappendiceal inflammatory stranding are statistically significant independent predictors for appendiceal perforation and are associated with increased hospital stay. PMID- 15749979 TI - Selective postsynaptic co-localization of MCT2 with AMPA receptor GluR2/3 subunits at excitatory synapses exhibiting AMPA receptor trafficking. AB - MCT2 is the main neuronal monocarboxylate transporter needed by neurons if they are to use lactate as an additional energy substrate. Previous evidence suggested that some MCT2 could be located in postsynaptic elements of glutamatergic synapses. Using post-embedding electron microscopic immunocytochemistry, it is demonstrated that MCT2 is present at postsynaptic density of asymmetric synapses, in the stratum radiatum of both rat hippocampal CA1 and CA3 regions, as well as at parallel fibre-Purkinje cell synapses in mouse cerebellum. MCT2 levels were significantly lower at mossy fibre synapses on CA3 neurons, and MCT2 was almost absent from symmetric synapses on CA1 pyramidal cells. It could also be demonstrated using quantitative double-labeling immunogold cytochemistry that MCT2 and AMPA receptor GluR2/3 subunits have a similar postsynaptic distribution at asymmetric synapses with high levels expressed within the postsynaptic density. In addition, as for AMPA receptors, a significant proportion of MCT2 is located on vesicular membranes within the postsynaptic spine, forming an intracellular pool available for a putative postsynaptic endo/exocytotic trafficking at these excitatory synapses. Altogether, the data presented provide evidence for MCT2 expression in the postsynaptic density area at specific subsets of glutamatergic synapses, and also suggest that MCT2, like AMPA receptors, could undergo membrane trafficking. PMID- 15749980 TI - Alpha phase synchronization predicts P1 and N1 latency and amplitude size. AB - The oscillatory phase-resetting model predicts that event-related potential (ERP) components are generated by a superposition of evoked oscillations with different frequencies. We investigate this question in a memory task in which human subjects had to retrieve a verbal label in response to the presentation of a picture. The results show that (i) evoked oscillations in the delta, theta, alpha and beta range undergo a significant phase resetting and (ii) become synchronized in absolute phase during small time windows that (iii) coincide with the latencies of the P1-N1 complex. Our conclusion is that the latencies of ERP components can be predicted at least in part by phase synchronization between frequencies. PMID- 15749981 TI - Use of a recombinant pseudorabies virus to analyze motor cortical reorganization after unilateral facial denervation. AB - A unilateral facial nerve injury (n7x) was found to influence the transcallosal spread of the attenuated strain of pseudorabies virus (PRV Bartha) from the affected (left) primary motor cortex (MI) to the contralateral MI of rats. We used Ba-DupLac, a recombinant PRV strain, for the tracing experiments since this virus was demonstrated to exhibit much more restricted transportation kinetics than that of PRV Bartha, and is therefore more suitable for studies of neuronal plasticity. Ba-Duplac injection primarily infected several neurons around the penetration channel, but hardly any transcallosally infected neurons were observed in the contraleral MI. In contrast, after right facial nerve injury, Ba DupLac was transported from the primarily infected neurons in the left MI to the contralateral side, and resulted in the labeling of several neurons due to a transneuronal infection. These results reveal that a peripheral nerve injury induces changes in the Ba-DupLac infection pattern in the related cortical areas. These findings and the literature data suggest that this phenomenon may be related to the changes in the expression or to the redistribution of cell adhesion molecules, which are known to facilitate the entrance and/or transmission of PRV into neurons. PMID- 15749982 TI - Activity-dependent regulation of synapse and dendritic spine morphology in developing barrel cortex requires phospholipase C-beta1 signalling. AB - The phospholipase C-beta1 (PLC-beta1) signalling pathway, activated via metabotropic glutamate receptors (mGluRs), is implicated in activity-dependent development of the cerebral cortex, as both PLC-beta1 and mGluR5 knockout mice exhibit disrupted barrel formation in somatosensory cortex. To characterize the effects of this signalling system on development of synaptic circuitry in barrel cortex, we have examined neuronal ultrastructure, synapse formation and dendritic spine morphology in PLC-beta1 knockout mice. Qualitative ultrastructure of neurons and synapse density in layers 2-4 of barrel cortex were unchanged in PLC beta1 knockout mice during development [postnatal day (P) 5] and in mature cortex (P19-21). We found a decrease in the proportion of synapses with symmetric morphology at P5 that was gone by P19-21, indicating a transient imbalance in excitatory and inhibitory circuitry. We also investigated dendritic spines by back-labelling layer 5 pyramidal neurons with carbocyanine. We observed normal dendritic spine densities on apical dendrites as they passed through layer 4 of barrel cortex, but spine morphology was altered in PLC-beta1 knockout mice at P9. These observations indicate that the PLC-beta1 signalling pathway plays a role in the development of normal cortical circuitry. Interrupting this regulation leads to changes in synapse and dendritic spine morphology, possibly altering post synaptic integration of signal. PMID- 15749984 TI - Degradation of signal timing in cortical areas V1 and V2 of senescent monkeys. AB - Senescence in monkeys results in a degradation of the functional properties of cortical cells as well as prolonged hyperactivity. We have now compared the spontaneous and visually evoked activity levels, as well as the visual response latencies of cells in cortical areas V1 and V2 of young and very old monkeys. We found that V1 cells within layer 4 exhibit normal latencies. In contrast, in other parts of V1 and throughout V2 hyperactivity in old monkeys is accompanied by dramatic delays in both the intracortical and intercortical transfer of information. Extrastriate cortex (area V2) is affected more severely than striate cortex (V1). Delayed information processing in cerebral cortex should contribute to the declines in cortical function that accompany old age. PMID- 15749983 TI - Brain networks associated with cognitive reserve in healthy young and old adults. AB - In order to understand the brain networks that mediate cognitive reserve, we explored the relationship between subjects' network expression during the performance of a memory test and an index of cognitive reserve. Using H2(15)O positron emission tomography, we imaged 17 healthy older subjects and 20 young adults while they performed a serial recognition memory task for nonsense shapes under two conditions: low demand, with a unique shape presented in each study trial; and titrated demand, with a study list size adjusted so that each subject recognized shapes at 75% accuracy. A factor score that summarized years of education, and scores on the NART and the WAIS-R Vocabulary subtest was used as an index of cognitive reserve. The scaled subprofile model was used to identify a set of functionally connected regions (or topography) that changed in expression across the two task conditions and was differentially expressed by the young and elderly subjects. The regions most active in this topography consisted of right hippocampus, posterior insula, thalamus, and right and left operculum; we found concomitant deactivation in right lingual gyrus, inferior parietal lobe and association cortex, left posterior cingulate, and right and left calcarine cortex. Young subjects with higher cognitive reserve showed increased expression of the topography across the two task conditions. Because this topography, which is responsive to increased task demands, was differentially expressed as a function of reserve level, it may represent a neural manifestation of innate or acquired reserve. In contrast, older subjects with higher cognitive reserve showed decreased expression of the topography across tasks. This suggests some functional reorganization of the network used by the young subjects. Thus, for the old subjects this topography may represent an altered, compensatory network that is used to maintain function in the face of age-related physiological changes. PMID- 15749985 TI - Increased action potential firing rates of layer 2/3 pyramidal cells in the prefrontal cortex are significantly related to cognitive performance in aged monkeys. AB - The neurobiological substrates of significant age-related deficits in higher cognitive abilities mediated by the prefrontal cortex (PFC) are unknown. To address this issue, whole-cell current-clamp recordings were used to compare the intrinsic membrane and action potential (AP) firing properties of layer 2/3 pyramidal cells in PFC slices from young and aged behaviorally characterized rhesus monkeys. Most aged subjects demonstrated impaired performance in Delayed Non-Match to Sample (DNMS) task acquisition, DNMS 2 min delay and the Delayed Recognition Span task. Resting membrane potential and membrane time constant did not differ in aged relative to young cells, but input resistance was significantly greater in aged cells. Single APs did not differ in terms of threshold, duration or rise time, but their amplitude and fall time were significantly decreased in aged cells. Repetitive AP firing rates were significantly increased in aged cells. Within the aged group, there was a U shaped quadratic relationship between firing rate and performance on each behavioral task. Subjects who displayed either low or very high firing rates exhibited poor performance, while those who displayed intermediate firing rates exhibited relatively good performance. These data indicate that an increase in AP firing rate may be responsible, in part, for age-related PFC dysfunction. PMID- 15749986 TI - Functional manipulations of acetylcholinesterase splice variants highlight alternative splicing contributions to murine neocortical development. AB - Proliferation and differentiation of mammalian central nervous system progenitor cells involve concertedly controlled transcriptional and alternative splicing modulations. Searching for the developmental implications of this programming, we manipulated specific acetylcholinesterase (AChE) splice variants in the embryonic mouse brain. In wild type mice, 'synaptic' AChE-S appeared in migrating neurons, whereas the C-terminus cleaved off the stress-induced AChE-R variant associated with migratory radial glial fibers. Antisense suppression of AChE-R reduced neuronal migration, allowing increased proliferation of progenitor cells. In contrast, transgenic overexpression of AChE-R was ineffective, whereas transgenic excess of enzymatically active AChE-S or inactive AChE-Sin suppressed progenitors proliferation alone or both proliferation and neuronal migration, respectively. Our findings attribute to alternative splicing events an interactive major role in neocortical development. PMID- 15749987 TI - The reentry hypothesis: the putative interaction of the frontal eye field, ventrolateral prefrontal cortex, and areas V4, IT for attention and eye movement. AB - Attention is known to play a key role in perception, including action selection, object recognition and memory. Despite findings revealing competitive interactions among cell populations, attention remains difficult to explain. The central purpose of this paper is to link up a large number of findings in a single computational approach. Our simulation results suggest that attention can be well explained on a network level involving many areas of the brain. We argue that attention is an emergent phenomenon that arises from reentry and competitive interactions. We hypothesize that guided visual search requires the usage of an object-specific template in prefrontal cortex to sensitize V4 and IT cells whose preferred stimuli match the target template. This induces a feature-specific bias and provides guidance for eye movements. Prior to an eye movement, a spatially organized reentry from occulomotor centers, specifically the movement cells of the frontal eye field, occurs and modulates the gain of V4 and IT cells. The processes involved are elucidated by quantitatively comparing the time course of simulated neural activity with experimental data. Using visual search tasks as an example, we provide clear and empirically testable predictions for the participation of IT, V4 and the frontal eye field in attention. Finally, we explain a possible physiological mechanism that can lead to non-flat search slopes as the result of a slow, parallel discrimination process. PMID- 15749988 TI - Localization of the GLYT1 glycine transporter at glutamatergic synapses in the rat brain. AB - In this study, we present evidence that a glycine transporter, GLYT1, is expressed in neurons and that it is associated with glutamatergic synapses. Despite the presence of GLYT1 mRNA in both glial cells and in glutamatergic neurons, previous studies have mainly localized GLYT1 immunoreactivity to glial cells in the caudal regions of the nervous system. However, using novel sequence specific antibodies, we have identified GLYT1 not only in glia, but also in neurons. The immunostaining of neuronal elements could best be appreciated in forebrain areas such as the neocortex or the hippocampus, and it was found in fibers, terminal boutons and in some dendrites. Double labeling confocal microscopy with the glutamatergic marker vGLUT1 revealed an enrichment of GLYT1 in a subpopulation of glutamatergic terminals. Moreover, through electron microscopy, we observed an enrichment of GLYT1 in both the presynaptic and the postsynaptic aspects of putative glutamatergic terminals that established asymmetric synapses. In addition, we demonstrated that GLYT1 was physically associated with the NMDA receptor in a biochemical assay. In conclusion, the close spatial association of GLYT1 and glutamatergic synapses strongly supports a role for this protein in neurotransmission mediated by NMDA receptors in the forebrain, and perhaps in other regions of the CNS. PMID- 15749989 TI - Functional architecture of retinotopy in visual association cortex of behaving monkey. AB - While the receptive field properties of single neurons in the inferior parietal cortex have been quantitatively described from numerous electrical measurements, the visual topography of area 7a and the adjacent dorsal prelunate area (DP) remains unknown. This lacuna may be a technical byproduct of the difficulty of reconstructing tens to hundreds of penetrations, or may be the result of varying functional retinotopic architectures. Intrinsic optical imaging, performed in behaving monkey for extended periods of time, was used to evaluate retinotopy simultaneously at multiple positions across the cortical surface. As electrical recordings through an implanted artificial dura are difficult, the measurement and quantification of retinotopy with long-term recordings was validated by imaging early visual cortex (areas V1 and V2). Retinotopic topography was found in each of the three other areas studied within a single day's experiment. However, the ventral portion of DP (DPv) had a retinotopic topography that varied from day to day, while the more dorsal aspects (DPd) exhibited consistent retinotopy. This suggests that the dorsal prelunate gyrus may consist of more than one visual area. The retinotopy of area 7a also varied from day to day. Possible mechanisms for this variability across days are discussed as well as its impact upon our understanding of the representation of extrapersonal space in the inferior parietal cortex. PMID- 15749990 TI - Proteomic analysis of somatic embryogenesis in Medicago truncatula. Explant cultures grown under 6-benzylaminopurine and 1-naphthaleneacetic acid treatments. AB - The Medicago truncatula line 2HA has a 500-fold greater capacity to regenerate plants in culture by somatic embryogenesis than wild-type Jemalong. We have compared proteomes of tissue cultures from leaf explants of these two lines. Both 2HA and Jemalong explants were grown on media containing the auxin 1 naphthaleneacetic acid and the cytokinin 6-benzylaminopurine. Proteins were extracted from the cultures at different time points (2, 5, and 8 weeks), separated by two-dimensional gel electrophoresis, and detected by silver staining. More than 2,000 proteins could be reproducibly resolved and detected on each gel. Statistical analysis showed that 54 protein spots were significantly (P < 0.05) changed in expression (accumulation) during the 8 weeks of culture, and most of these spots were extracted from colloidal Coomassie-stained two dimensional gel electrophoresis gels and were subjected to matrix-assisted laser desorption ionization time-of-flight mass spectrometry or liquid chromatography tandem mass spectrometry analysis. Using a publicly available expressed sequence tag database and the Mascot search engine, we were able to identify 16 differentially expressed proteins. More than 60% of the differentially expressed protein spots had very different patterns of gene expression between 2HA and Jemalong during the 8 weeks of culture. PMID- 15749991 TI - Lotus japonicus metabolic profiling. Development of gas chromatography-mass spectrometry resources for the study of plant-microbe interactions. AB - Symbiotic nitrogen fixation (SNF) in legume root nodules requires differentiation and integration of both plant and bacterial metabolism. Classical approaches of biochemistry, molecular biology, and genetics have revealed many aspects of primary metabolism in legume nodules that underpin SNF. Functional genomics approaches, especially transcriptomics and proteomics, are beginning to provide a more holistic picture of the metabolic potential of nodules in model legumes like Medicago truncatula and Lotus japonicus. To extend these approaches, we have established protocols for nonbiased measurement and analysis of hundreds of metabolites from L. japonicus, using gas chromatography coupled with mass spectrometry. Following creation of mass spectral tag libraries, which represent both known and unknown metabolites, we measured and compared relative metabolite levels in nodules, roots, leaves, and flowers of symbiotic plants. Principal component analysis of the data revealed distinct metabolic phenotypes for the different organs and led to the identification of marker metabolites for each. Metabolites that were enriched in nodules included: octadecanoic acid, asparagine, glutamate, homoserine, cysteine, putrescine, mannitol, threonic acid, gluconic acid, glyceric acid-3-P, and glycerol-3-P. Hierarchical cluster analysis enabled discrimination of 10 groups of metabolites, based on distribution patterns in diverse Lotus organs. The resources and tools described here, together with ongoing efforts in the areas of genome sequencing, and transcriptome and proteome analysis of L. japonicus and Mesorhizobium loti, should lead to a better understanding of nodule metabolism that underpins SNF. PMID- 15749993 TI - Expression of nigrostriatal alpha 6-containing nicotinic acetylcholine receptors is selectively reduced, but not eliminated, by beta 3 subunit gene deletion. AB - mRNAs for the neuronal nicotinic acetylcholine receptor (nAChR) alpha6 and beta3 subunits are abundantly expressed and colocalized in dopaminergic cells of the substantia nigra and ventral tegmental area. Studies using subunit-null mutant mice have shown that alpha6- or beta3-dependent nAChRs bind alpha-conotoxin MII (alpha-CtxMII) with high affinity and modulate striatal dopamine release. This study explores the effects of beta3 subunit-null mutation on striatal and midbrain nAChR expression, composition, and pharmacology. Ligand binding and immunoprecipitation experiments using subunit-specific antibodies indicated that beta3-null mutation selectively reduced striatal alpha6* nAChR expression by 76% versus beta3(+/+) control. Parallel experiments showed a smaller reduction in both midbrain alpha3* and alpha6* nAChRs (34 and 42% versus beta3(+/+) control, respectively). Sedimentation coefficient determinations indicated that residual alpha6* nAChRs in beta3(-/-) striatum were pentameric, like their wild-type counterparts. Immunoprecipitation experiments on immunopurified beta3* nAChRs demonstrated that almost all wild-type striatal beta3* nAChRs also contain alpha4, alpha6, and beta2 subunits, although a small population of non-beta3 alpha6* nAChRs is also expressed. beta3 subunit incorporation seemed to increase alpha4 participation in alpha6beta2* complexes. (125)I-Epibatidine competition binding studies showed that the alpha-CtxMII affinity of alpha6* nAChRs from the striata of beta3(-/-) mice was similar to those isolated from beta3(+/+) animals. Together, the results of these experiments show that the beta3 subunit is important for the correct assembly, stability and/or transport of alpha6* nAChRs in dopaminergic neurons and influences their subunit composition. However, beta3 subunit expression is not essential for the expression of alpha6*, high-affinity alpha-CtxMII binding nAChRs. PMID- 15749992 TI - The mycorrhizal fungus Gigaspora margarita possesses a CuZn superoxide dismutase that is up-regulated during symbiosis with legume hosts. AB - A full-length cDNA showing high similarity to previously described CuZn superoxide dismutases (SODs) was identified in an expressed sequence tag collection from germinated spores of the arbuscular mycorrhizal fungus Gigaspora margarita (BEG 34). The corresponding gene sequence, named GmarCuZnSOD, is composed of four exons. As revealed by heterologous complementation assays in a yeast mutant, GmarCuZnSOD encodes a functional polypeptide able to confer increased tolerance to oxidative stress. The GmarCuZnSOD RNA was differentially expressed during the fungal life cycle; highest transcript levels were found in fungal structures inside the roots as observed on two host plants, Lotus japonicus and Medicago truncatula. These structures also reacted positively to 3,3'-diaminobenzidine, used to localize H2O2 accumulation. This H2O2 is likely to be produced by CuZnSOD activity since treatment with a chelator of copper ions, generally used to inhibit CuZnSODs, strongly reduced the 3,3'-diaminobenzidine deposits. A slight induction of GmarCuZnSOD gene expression was also observed in germinated spores exposed to L. japonicus root exudates, although the response showed variation in independent samples. These results provide evidence of the occurrence, in an arbuscular mycorrhizal fungus, of a functional SOD gene that is modulated during the life cycle and may offer protection as a reactive oxygen species-inactivating system against localized host defense responses raised in arbuscule-containing cells. PMID- 15749995 TI - Chemically distinct ligands promote differential CB1 cannabinoid receptor-Gi protein interactions. AB - To understand how structurally distinct ligands regulate CB(1) receptor interactions with Gi1, Gi2, and Gi3, we quantified the Galphai and betagamma proteins that coimmunoprecipitate with the CB(1) receptor from a detergent extract of N18TG2 membranes in the presence of ligands. A mixture of A, R, G(GDP) (or G_), and ARG(GDP) (or ARG_) complexes was observed in the presence of aminoalkylindole (R)-(+)-[2,3-dihydro-5-methyl-3-(4 morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone (WIN 55,212-2) for all three RGalphai complexes, cannabinoid desacetyllevonantradol for Galphai1 and Galphai2, and eicosanoid (R) methanandamide for Galphai3. Desacetyllevonantradol maintained RGalphai3 complexes and (R)-methanandamide maintained RGalphai1 and RGalphai2 complexes even in the presence of a nonhydrolyzable GTP analog. The biaryl pyrazole antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl 1H-pyrazole-3-carboximide hydrochloride (SR141716) maintained all three RGalphai complexes. Gbeta proteins, and to a certain extent Ggamma2, exhibited the same association/dissociation pattern as the Galpha proteins. A GDP analog had no influence on any of these association/dissociation reactions and failed to promote sequestration of G proteins. These results can be explained by invoking the existence of an inverse agonist-supported inactive state in the ternary complex equilibrium model. WIN 55,212-2 behaves as an agonist for all three Gi subtypes; SR141716 behaves as an inverse agonist for all three Gi subtypes; desacetyllevonantradol behaves as an agonist for Gi1 and Gi2, and an inverse agonist at Gi3; and (R)-methanandamide behaves as an inverse agonist at Gi1 and Gi2, and an agonist at Gi3. These ligand-selective G protein responses imply that multiple conformations of the receptor could be evoked by ligands to regulate individual G proteins. PMID- 15749996 TI - 14th workshop on fetal cells and fetal DNA. PMID- 15749994 TI - Breast cancer resistance protein (Bcrp1/Abcg2) limits net intestinal uptake of quercetin in rats by facilitating apical efflux of glucuronides. AB - The intestinal absorption of the flavonoid quercetin in rats is limited by the secretion of glucuronidated metabolites back into the gut lumen. The objective of this study was to determine the role of the intestinal efflux transporters breast cancer resistance protein (Bcrp1)/Abcg2 and multidrug resistance-associated protein 2 (Mrp2)/Abcc2. To study the possible involvement of Mrp2, we compared intestinal uptake of quercetin-3-glucoside between control and Mrp2-deficient rats, using an in situ intestinal perfusion system. The contribution of Bcrp1 was determined using the specific inhibitor fumitremorgin C (FTC) in Mrp2-deficient rats. Furthermore, vectorial transport of quercetin was studied in Madin-Darby canine kidney (MDCK)II cells transfected with either human MRP2 or murine Bcrp1. In these MDCKII cells, we showed an efficient efflux-directed transport of quercetin by mouse Bcrp1, whereas in control and MRP2-transfected cells no vectorial transport of quercetin was observed. In Mrp2-deficient rats, intestinal uptake of quercetin from quercetin-3-glucoside, efflux of quercetin glucuronides to the gut lumen, and plasma concentration of quercetin were similar to that in control rats. When intestinal Bcrp1 was inhibited by FTC in Mrp2-deficient rats, total plasma concentrations of quercetin and its methylated metabolite isorhamnetin after 30 min of perfusion were more than twice that of controls (12.3 +/- 1.5 versus 5.6 +/- 1.3 muM; p < 0.01), whereas uptake of free quercetin from the intestinal lumen was not affected. Instead, inhibition of Bcrp1 lowered the efflux of quercetin glucuronides into the perfusion fluid by approximately 4 fold. In conclusion, Bcrp1 limits net intestinal absorption of quercetin by pumping quercetin glucuronides back into the lumen. PMID- 15749997 TI - Preimplantation genetic diagnosis--an overview. AB - Since the early 1990s, preimplantation genetic diagnosis (PGD) has been expanding in scope and applications. Selection of female embryos to avoid X-linked disease was carried out first by polymerase chain reaction, then by fluorescence in situ hybridization (FISH), and an ever-increasing number of tests for monogenic diseases have been developed. Couples with chromosome rearrangements such as Robertsonian and reciprocal translocations form a large referral group for most PGD centers and present a special challenge, due to the large number of genetically unbalanced embryos generated by meiotic segregation. Early protocols used blastomeres biopsied from cleavage-stage embryos; testing of first and second polar bodies is now a routine alternative, and blastocyst biopsy can also be used. More recently, the technology has been harnessed to provide PGD-AS, or aneuploidy screening. FISH probes specific for chromosomes commonly found to be aneuploid in early pregnancy loss are used to test blastomeres for aneuploidy, with the aim of replacing euploid embryos and increasing pregnancy rates in groups of women who have poor IVF success rates. More recent application of PGD to areas such as HLA typing and social sex selection have stoked public controversy and concern, while provoking interesting ethical debates and keeping PGD firmly in the public eye. PMID- 15749998 TI - Sperm and blastomere aneuploidy detection in reproductive genetics and medicine. AB - The use of multiple probes in fluorescence in situ hybridization (FISH) permits the simultaneous analysis of several chromosomes in both blastomeres and spermatozoa. Preimplantation genetic diagnosis (PGD) for aneuploidy provides information on embryonic chromosomal status, enabling the selection of embryos carrying aneuploid condition. This strategy directly affects implantation, as documented for patients with a poor prognosis for pregnancy, who have the tendency to generate high proportions of chromosomally abnormal embryos. PGD for aneuploidy also has contributed information on early phases in human embryology by clarifying the molecular basis in some cases of irregular development. Multicolor FISH has also been used to study chromosomes on spermatozoa. Experimental strategies and modifications enabled the analysis of samples with a very low number of sperm cells, including samples retrieved from the genital tract or directly from the testicular tissue. The results confirmed that the incidence of aneuploidy increases proportionally with the severity of the male factor condition. This observation suggests that, in selected cases, the paternal contribution to aneuploidy in the developing conceptus could be more relevant than expected from general data from aborted fetuses and live births. PMID- 15749999 TI - Fluorescence in situ hybridization and spectral imaging analysis of human oocytes and first polar bodies. AB - We investigated the frequencies of abnormalities involving either chromosome 1, 16, 18, or 21 in failed-fertilized human oocytes. Although abnormalities involving chromosome 16 showed an age-dependent increase, results for the other chromosomes did not show statistically significant differences among the three age groups, <35 years, 35-39 years, and >39 years. The scoring of four chromosomes is likely to underestimate the true rate of aneuploid cells. Therefore, for a pilot study investigating a more-comprehensive analysis of oocytes and their corresponding first polar bodies, we developed a novel eight probe chromosome enumeration scheme using fluorescence in situ hybridization and spectral imaging analysis. PMID- 15750000 TI - Topology of chromosomes 18 and X in human blastomeres from 3- to 4-day-old embryos. AB - The positions of chromosomes 18 and X fluorescence in situ hybridization signals were analyzed in blastomeres generated from human in vitro fertilization 3- to 4 day-old embryos after preimplantation screening of aneuploidy of chromosomes 13, 16, 18, 21, 22, X, and Y. Fluorescent signal localization compared with a three dimensional sphere model of random signal distribution revealed significant differences, providing evidence of peripheral localization of chromosome 18 in aneuploid (p=0.0013) and aneuploid/euploid blastomeres (p=0.0011). No differences were found in localization of chromosome 18 in euploid and in chromosome X in euploid and aneuploid blastomeres. PMID- 15750001 TI - Polar body diagnosis for hemophilia a using multiplex PCR for linked polymorphic markers. AB - Preimplantation genetic diagnosis (PGD) is usually performed on blastomeres. In Germany, the only possibility to perform PGD is by analysis of polar bodies. We performed PGD using polar bodies in a woman who is a carrier of hemophilia A. Multiplex PCR followed by nested fluorescent PCR for five linked polymorphic markers was established. From 11 analyzed polar bodies, only 1 showed alleles linked to the mutation. The corresponding oocyte was transferred and no pregnancy was established. As seen in other investigations, the rate of heterozygous first polar bodies is surprisingly high. PMID- 15750002 TI - First-trimester screening: an overview. AB - An improvement in prenatal screening for chromosomal defects has been achieved by combining sonography and biochemical markers. Analyzing markers taken from maternal blood such as pregnancy-associated plasma protein A and free beta-human chorionic gonadotropin in combination with the ultrasound marker nuchal translucency provides detection rates of 90% for the most important chromosomal anomalies. In addition, nuchal translucency is a marker for severe heart defects. This report discusses the potential of new markers such as the nasal bone. PMID- 15750003 TI - Rapid prenatal diagnosis of aneuploidy using quantitative fluorescence-PCR (QF PCR). AB - Molecular cytogenetic aneuploidy testing for pregnant women at increased risk of chromosome abnormality leads to rapid reassurance for those with normal results and earlier decisions on pregnancy management in the case of abnormality. We tested 9080 prenatal samples using a one-tube QF-PCR test for trisomies 13, 18, and 21; the abnormality rate was 5.9%. There were no misdiagnoses for non-mosaic trisomy. A sex chromosome multiplex was developed that detects structural sex chromosome abnormalities as well as aneuploidies. The sex chromosome test was targeted at pregnancies (272) with specific abnormalities suggestive of Turner syndrome; 13.2% showed 45,X, confirmed by follow-up analysis. PMID- 15750004 TI - Rapid prenatal diagnostics in the interphase nucleus: procedure and cut-off rates. AB - Results of a rapid prenatal aneuploidy screening in uncultured amniocytes using FISH are available within 24 hr. However, care has to be taken of possible pitfalls in connection with the commercially available probe sets and in the interpretation of results in general. Here we present our cut-off rates for and the way in which we handle the Aneu Vysion kit (ABBOTT/Vysis), based on the experience of 1200 studied cases. In summary, the method is reliable, and the risk for misdiagnosis is low ( approximately 0.4%). Nonetheless, the test should be used exclusively as a preamble to full chromosome analysis by microscopy. PMID- 15750005 TI - Recent advances in fetal nucleic acids in maternal plasma. AB - The discovery of cell-free fetal DNA in maternal plasma in 1997 has opened up new possibilities for noninvasive prenatal diagnosis. Circulating fetal DNA molecules have been detected in maternal plasma from the first trimester onwards and can be robustly detected using a variety of molecular methods. This approach has been used for the prenatal investigation of sex-linked diseases, fetal RhD status, and prenatal exclusion of beta-thalassemia major. Recently, fetal RNA has also been found in maternal plasma. Such fetal RNA has been shown to originate from the placenta and to be remarkably stable. The use of microarray-based approaches has made it feasible to rapidly generate new circulating RNA markers. It is hoped that further developments in this field will make the routine and widespread practice of noninvasive nucleic acid-based prenatal diagnosis for common pregnancy-associated disorders feasible in the near future. PMID- 15750006 TI - Analysis of cell-free fetal DNA in plasma and serum of pregnant women. AB - Sixty blood samples from pregnant women during gestational weeks 9-28 were investigated. Cell-free fetal DNA was extracted from maternal plasma or serum to be detected by nested PCR for determination of fetal gender. The SRY gene as a marker for fetal Y chromosome was detected in 34/36 women carrying a male fetus. In 3/24 women carrying female fetuses, the SRY sequence was also detected. Overall, fetal sex was correctly predicted in 91.7% of the cases. Therefore, the new, non-invasive method of prenatal diagnosis of fetal gender for women at risk of producing children with X-linked disorders is reliable, secure, and can substantially reduce invasive prenatal tests. PMID- 15750007 TI - Non-invasive fetal RHD and RHCE genotyping using real-time PCR testing of maternal plasma in RhD-negative pregnancies. AB - We assessed the feasibility of fetal RHD and RHCE genotyping by analysis of DNA extracted from plasma samples of RhD-negative pregnant women using real-time PCR and primers and probes targeted toward RHD and RHCE genes. We analyzed 45 pregnant women in the 11th to 40th weeks of pregnancy and correlated the results with serological analysis of cord blood after delivery. Non-invasive prenatal fetal RHD exon 7, RHD exon 10, RHCE exon 2 (C allele), and RHCE exon 5 (E allele) genotyping analysis of maternal plasma samples was correctly performed in 45 out of 45 RhD-negative pregnant women delivering 24 RhD-, 17 RhC-, and 7 RhE-positive newborns. Detection of fetal RHD and the C and E alleles of RHCE gene from maternal plasma is highly accurate and enables implementation into clinical routine. We recommend performing fetal RHD and RHCE genotyping together with fetal sex determination in alloimmunized D-negative pregnancies at risk of hemolytic disease of the newborn. In case of D-negative fetus, amplification of another paternally inherited allele (SRY and/or RhC and/or RhE positivity) proves the presence of fetal DNA in maternal circulation. PMID- 15750008 TI - Application of fetal DNA detection in maternal plasma: a prenatal diagnosis unit experience. AB - Non-invasive prenatal diagnosis tests based on the analysis of fetal DNA in maternal plasma have potential to be a safer alternative to invasive methods. So far, different studies have shown mainly fetal sex, fetal RhD, and quantitative variations of fetal DNA during gestation with fetal chromosomal anomalies or gestations at risk for preeclampsia. The objective of our research was to evaluate the use of fetal DNA in maternal plasma for clinical application. In our study, we have established the methodology needed for the analysis of fetal DNA. Different methods were used, according to the requirements of the assay. We have used quantitative fluorescent polymerase chain reaction (QF-PCR) to perform fetal sex detection with 90% sensitivity. The same technique permitted the detection of fetal DNA from the 10th week of gestation to hours after delivery. We have successfully carried out the diagnosis of two inherited disorders, cystic fibrosis (conventional PCR and restriction analysis) and Huntington disease (QF PCR). Ninety percent of the cases studied for fetal RhD by real-time PCR were correctly diagnosed. The detection of fetal DNA sequences is a reality and could reduce the risk of invasive techniques for certain fetal disorders in the near future. PMID- 15750009 TI - First-trimester NRBC count in maternal circulation: correlation with doppler ultrasound studies. AB - This study aimed to determine whether the number of nucleated red blood cells (NRBCs) in maternal circulation during the first trimester of pregnancy could identify pregnancies that will have an anomalous Doppler in the second trimester. A total of 85 blood samples were obtained at 11-14 weeks of gestation with mean uterine arterial perfusion index >1.6, as noted by Doppler ultrasonography. NRBCs were enriched by magnetic automated cell sorting using anti-CD71 and were stained with May/Grunwald/Giemsa. A total of 4.8 NRBCs (range 1-75) were identified in 68 cases. Follow-up scans at 22-24 weeks were available in 46 cases. In 39 women, blood flow in the uterine arteries normalized, whereas in seven, high resistance was noted. One woman in the high-resistance group developed preeclampsia (PET; four NRBCs) and another delivered an intrauterine growth restriction (IUGR) baby (75 NRBCs). The number of NRBCs in women whose Doppler indices later normalized and in those who continued to have increased impedance was similar. The study indicates that NRBC number in maternal circulation during the first trimester cannot be used to screen pregnancies at high risk for developing preeclampsia (PET)/IUGR. High-impedance blood flow in the uterine arteries in the first trimester may be due to an unfinished process of trophoblastic invasion, most likely to be completed successfully by 22-24 weeks. PMID- 15750010 TI - FISH analysis of all fetal nucleated cells in maternal whole blood: improved specificity by the use of two Y-chromosome probes. AB - Current cytogenetic approaches in noninvasive prenatal diagnosis focus on fetal nucleated red blood cells in maternal blood. This practice may be too restrictive because a vast proportion of other fetal cells is ignored. Recent studies have indicated that fetal cells can be directly detected, without prior enrichment, in maternal blood samples by fluorescence in situ hybridization (FISH) analysis for chromosomes X and Y (XY-FISH). In our blinded analysis of 40 maternal blood samples, we therefore examined all fetal cells without any enrichment. Initial examinations using conventional XY-FISH indicated a low specificity of 69.4%, which could be improved to 89.5% by the use of two different Y-chromosome specific probes (YY-FISH) with only a slight concomitant decrease in sensitivity (52.4% vs 42.9%). On average, 12-20 male fetal cells/ml of maternal blood were identified by XY- and YY-FISH, respectively. PMID- 15750011 TI - Fetal gender determination and BclI polymorphism using nucleated erythrocytes in maternal blood. AB - This study demonstrated determination of fetal gender from nucleated red blood cells (NRBCs) in maternal blood and attempted to apply prenatal diagnosis of hemophilia A using BclI DNA polymorphism. Venous blood was drawn from 20 pregnant women, and NRBCs were recovered by magnetic activated cell sorting and anti-GPA (glycophorin A) immunostaining. After microdissector isolation of the NRBCs, primer extension preamplification (PEP) and nested PCR of the amelogenin gene were performed to determine fetal gender. We also performed PEP and nested PCR of BclI polymorphism to verify the validity of prenatal diagnosis of hemophilia A. DNA amplification was achieved in 107 cells (51.9%) and fetal gender determined with 65.0% accuracy. Unfortunately, we could not verify the validity within the scope of this study. However, in a larger number of cases that are informative in BclI polymorphism, we will be able to identify patients affected by hemophilia A using fetal NRBCs in maternal blood. PMID- 15750012 TI - Evaluation of a soybean lectin-based method for the enrichment of erythroblasts. AB - We performed a comparative study of the enrichment of erythroblasts by a soybean agglutinin galactose-specific lectin method and a standardized magnetic cell sorting (MACS) protocol. Blood samples, obtained from 11 pregnant women at between 11 and 40 weeks of gestation, were split and examined by each method in parallel. The number of erythroblasts recovered by the lectin method was approximately eightfold higher than the number obtained by MACS. Our data suggest that the lectin-based method may provide a better approach for the enrichment of rare fetal erythroblasts from maternal blood. PMID- 15750013 TI - The fetal erythroblast is not the optimal target for non-invasive prenatal diagnosis: preliminary results. AB - Fetal cells, present in the blood of pregnant women, are potential targets for non-invasive prenatal diagnosis. The fetal erythroblast has been the favorite target cell type. We investigated four methods of enrichment for fetal erythroblasts, identifying only three fetal erythroblasts in 573 ml of maternal blood. This is much less than the expected two to six fetal cells per ml of maternal blood. Hamada and Krabchi used a cell type-independent marker, i.e., the Y chromosome in maternal blood from male pregnancies after Carnoy fixation, leaving the nuclei for hybridization with X-and Y-chromosome-specific probes. We found with a similar technique 28 fetal cells in 15 ml of maternal blood. The fetal origin of cells was confirmed by hybridizing the nuclei with X- and Y chromosome-specific probes, using two consecutive hybridizations with the two probes in opposite colors (reverse FISH). Candidate fetal cells were inspected after each hybridization. Only cells that were found to change the color of both probe signals from first to second hybridization were diagnosed as fetal. To reduce the labor-intensive slide screening load, we used semiautomated scanning microscopy to search for candidate cells. We conclude that erythroblasts form only a small fraction of fetal cells present in maternal blood. PMID- 15750014 TI - Trophoblasts isolated from the maternal circulation: in vitro expansion and potential application in non-invasive prenatal diagnosis. AB - Prenatal diagnosis based on rare fetal cells in maternal blood is currently not a feasible option. An effort was made to improve cell yields by targeting trophoblast cells. After sorting, the HLA-G-positive cell fraction was analyzed directly or after culture. In situ hybridization technology was applied to prove fetal cell source in samples from women carrying a male fetus and to predict gender in samples without previous knowledge of fetal sex. In vitro culture led to a significant increase in fetal cells and accurate gender prediction in 93% of these samples. This approach might be useful for non-invasive prenatal diagnosis. PMID- 15750015 TI - Proteome analysis of maternal serum samples for trisomy 21 pregnancies using ProteinChip arrays and bioinformatics. AB - A surface-enhanced laser desorption/ionization time of flight (SELDI-TOF)-based ProteinChip System was used as a tool for rapid discovery and identification of protein patterns in serum that discriminate between trisomy 21 and unaffected pregnancies. We analyzed 24 serum samples from women carrying a trisomy 21 pregnancy and 32 with an unaffected pregnancy, ranging from 10.0 to 14.0 weeks of gestation. The resulting protein profiles were submitted to a clustering algorithm, a rule extraction, a rating, and a rule base construction step. For the generated combined rule base, the specificity and sensitivity for the prediction of a trisomy 21 pregnancy reach 97% and 91%, respectively. PMID- 15750016 TI - A monoclonal antibody with potential for aiding non-invasive prenatal diagnosis: utility in screening of pregnant women at risk of preeclampsia. AB - The development of a non-invasive method of prenatal diagnosis in maternal blood has been the goal of our investigations during the last years. We have developed several anti-CD71 monoclonal antibodies and optimized a protocol for the isolation of nucleated red blood cells (NRBC) from peripheral maternal blood. The enhanced traffic of fetal erythroblasts into the maternal circulation in preeclampsia has been investigated by several groups. In this study, we compared one of our antibodies, 2F6.3, with a commercial anti-CD71 antibody in blood samples from pregnant women suffering pregnancy-induced hypertension (PIH) and in a control group of pregnant women without clinical features suggestive of an increased risk of developing preeclampsia. The mAb 2F6.3, developed by our group, has succeeded in isolating a significantly higher number of erythroblasts with less maternal cell contamination than the commercial antibody in both women with PIH and in the control group (p<0.01; Wilcoxon Signed Ranks Test). Fluorescence in situ hybridization analysis also demonstrated that 2F6.3 is a better antibody for the isolation of fetal NRBC in maternal blood than the commercial anti-CD71 antibody. PMID- 15750017 TI - Prenatal diagnosis and molecular cytogenetic characterization of an unusual complex structural rearrangement in a pregnancy following intracytoplasmic sperm injection (ICSI). AB - We report on a balanced complex chromosomal aberration detected in a fetus after amniocentesis. The pregnancy was achieved after intracytoplasmic sperm injection. GTG-banding revealed a complex structurally rearranged karyotype with a translocation between chromosomes 5 and 15 and an additional paracentric inversion in the der(15) between bands 5q11.2 and 5q15. Ag-NOR staining showed an interstitial active nuclear organizer region in the der(15). Molecular cytogenetic analyses using whole-chromosome-painting probes, comparative genomic hybridization, and multicolor banding did not point to further structural aberrations or imbalances. Therefore, a complex rearrangement with three breakpoints has occurred, and the karyotype can be described as 46,XX,der(5)t(5;15) (q11.2;p12),der(15)t(5;15)(q11.2;p12)inv(5)(q11.2q15). PMID- 15750018 TI - A highly complex chromosomal rearrangement between five chromosomes in a healthy female diagnosed in preparation for intracytoplasmatic sperm injection. AB - We report a case of a de novo complex chromosomal rearrangement among five chromosomes found in a clinically healthy woman. The only indication for chromosome analysis was a planned intracytoplasmatic sperm injection. Physical examination, including internal and external genitals, and ovaries and hormone status were normal. Banding cytogenetics showed a rearrangement among chromosomes #3, #4, #7, #9, and #17. Twenty-four-color fluorescence in situ hybridization and multicolor banding were applied to characterize the translocations and breakpoints more precisely. This confirmed the involved chromosomes and revealed two breakpoints in chromosome #4. This six-breakpoint rearrangement [der(3)t(3;4), der(4)t(17;4;7), der(7)t(3;7), der(9)t(4;9), and der(17)t(9;17)] seemed to be balanced on a molecular cytogenetic level, although submicroscopic deletions or duplications close to the breakpoints cannot be excluded. PMID- 15750019 TI - Three cases with enlarged acrocentric p-arms and two cases with cryptic partial trisomies. AB - In three cases, banding analysis revealed a normal karyotype except for an enlarged short arm of one chromosome 13 or 15. To clarify whether this enlargement was due to a heteromorphism or to a cryptic chromosomal trisomy, so called cenM-FISH probe sets containing a microdissection-derived probe specific for the acrocentric human p-arms were applied. The results enabled us to confirm in one case and to exclude in two cases that the enlargement on the suspect chromosome was due to a p-arm polymorphism. M-FISH and/or microdissection were used to resolve the nature of the rearrangements, i.e., partial trisomies 6 and 19. PMID- 15750021 TI - Dup(13)(q14.2-q14.3): yet another new differential diagnostic aspect for short stature-like phenotype. AB - We report on the case of a pregnant woman with hyposomia who was previously suspected of having Turner syndrome. Prenatal cytogenetic diagnostics showed a fetal karyotype of 46,XX,dup(13)(q14.2q21.1) ish.13q14(RB1 x 3). Parental and grandparental chromosome analyses were performed and the dup(13) was found to be of maternal origin (de novo). The pregnancy was continued and a healthy female child was born with normal development apart from growth retardation. The reported chromosomal aberration is, together with two other cases reported in the literature, the first hint of a short stature-like phenotype due to dup(13)(q14.2q14.3). PMID- 15750020 TI - Pallister-Killian syndrome: rapid decrease of isochromosome 12p frequency during amniocyte subculturing. Conclusion for strategy of prenatal cytogenetic diagnostics. AB - Pallister-Killian syndrome (PKS) is characterized cytogenetically by mosaic tetrasomy of chromosome 12p. Routine prenatal diagnosis of PKS is still complicated because of the difficulties of discriminating between the supernumerary isochromosome 12p and the duplication 21q and because of the variable level of mosaicism. The frequency of cells with an extra metacentric chromosome i(12)(p10) is usually determined by tissue-limited or tissue-specific mosaicism. We demonstrated a decrease of the abnormal clone with extra i(12p) in the amniotic fluid cells of the PKS fetus during amniocyte subculturing. The rapid loss of the i(12p) in the course of amniocyte subculturing should be the focus of attention during prenatal karyotyping. This is especially necessary for cultures with slow growth, which require further interpretation of the result during cytogenetic diagnosis of PKS. PMID- 15750022 TI - Another small supernumerary marker chromosome (sSMC) derived from chromosome 2: towards a genotype/phenotype correlation. AB - Here we report a prenatally detected small supernumerary marker chromosome (sSMC) derived from chromosome 2 as demonstrated by cenM-FISH (centromere-specific multicolor fluorescence in situ hybridization). By application of a recently described subcentromere-specific probe set (subcenM-FISH) for chromosome 2, the presence of a small partial trisomy due to a karyotype 47,XX,+r(2)(::p11.1 >q11.2::) was demonstrated. Including this case, a total of 11 patients with sSMC(2) are described throughout the literature. Based on that data, a first genotype/phenotype correlation according to the size and structure of the marker is suggested. PMID- 15750023 TI - QF-PCR examination of parental and meiotic origin of trisomy 21 in Central and Eastern Europe. AB - Study of parental/meiotic origin of free trisomy 21 in nuclear families from Russia (70 cases), Ukraine (32 cases), and 22 from Germany revealed maternal nondisjunction in 77.3% (Germany), 93.8% (Ukraine), and 91.4% (Russia), paternal origin in 13.6%, 6.2%, and 8.6%, respectively. Maternal meiosis I errors were found in 84.4% (Ukraine), 77.1% (Russia), paternal origin in 3.1% (Ukraine), 2.9% (Russia). Maternal meiosis II errors occurred in 9.4% and 14.3% and paternal in 3.1% and 5.7% in Ukraine and Russia, respectively. No significant differences were found in maternal/paternal origin among Ukraine, Russia, Germany, and published data from other European regions. PMID- 15750024 TI - Evidence for high frequency of chromosomal mosaicism in spontaneous abortions revealed by interphase FISH analysis. AB - Numerical chromosomal imbalances are a common feature of spontaneous abortions. However, the incidence of mosaic forms of chromosomal abnormalities has not been evaluated. We have applied interphase multicolor fluorescence in situ hybridization using original DNA probes for chromosomes 1, 9, 13, 14, 15, 16, 18, 21, 22, X, and Y to study chromosomal abnormalities in 148 specimens of spontaneous abortions. We have detected chromosomal abnormalities in 89/148 (60.1%) of specimens. Among them, aneuploidy was detected in 74 samples (83.1%). In the remaining samples, polyploidy was detected. The mosaic forms of chromosome abnormality, including autosomal and sex chromosomal aneuploidies and polyploidy (31 and 12 cases, respectively), were observed in 43/89 (48.3%) of specimens. The most frequent mosaic form of aneuploidy was related to chromosome X (19 cases). The frequency of mosaic forms of chromosomal abnormalities in samples with male chromosomal complement was 50% (16/32 chromosomally abnormal), and in samples with female chromosomal complement, it was 47.4% (27/57 chromosomally abnormal). The present study demonstrates that the postzygotic or mitotic errors leading to chromosomal mosaicism in spontaneous abortions are more frequent than previously suspected. Chromosomal mosaicism may contribute significantly to both pregnancy complications and spontaneous fetal loss. PMID- 15750025 TI - First systematic CGH-based analyses of ancient DNA samples of malformed fetuses preserved in the Meckel Anatomical Collection in Halle/Saale (Germany). AB - We present the first data on our comparative genomic hybridization (CGH)-based strategy for the analysis of ancient DNA (aDNA) samples extracted from fetuses preserved in the Meckel Anatomical Collection in Halle, Germany. The collection contains numerous differently fixed ancient samples of fetal malformations collected from the middle of the 18th to the early 19th century. The main objective of this study is to establish a "standard" aDNA extraction and amplification protocol as a prerequisite for successful CGH analyses to detect or exclude chromosomal imbalances possibly causative for the malformations described for the fetuses. PMID- 15750026 TI - The variation of aneuploidy frequency in the developing and adult human brain revealed by an interphase FISH study. AB - Despite the lack of direct cytogenetic studies, the neuronal cells of the normal human brain have been postulated to contain normal (diploid) chromosomal complement. Direct proof of a chromosomal mutation presence leading to large scale genomic alterations in neuronal cells has been missing in the human brain. Large-scale genomic variations due to chromosomal complement instability in developing neuronal cells may lead to the variable level of chromosomal mosaicism probably having a substantial effect on brain development. The aim of the present study was the pilot assessment of chromosome complement variations in neuronal cells of developing and adult human brain tissues using interphase multicolor fluorescence in situ hybridization (mFISH). Chromosome-enumerating DNA probes from the original collection (chromosomes 1, 13 and 21, 18, X, and Y) were used for the present pilot FISH study. As a source of fetal brain tissue, the medulla oblongata was used. FISH studies were performed using uncultured fetal brain samples as well as organotypic cultures of medulla oblongata tissue. Cortex tissues of postmortem adult brain samples (Brodmann area 10) were also studied. In cultured in vitro embryonic neuronal brain cells, an increased level of aneuploidy was found (mean rate in the range of 1.3-7.0% per individual chromosome, in contrast to 0.6-3.0% and 0.1-0.8% in uncultured fetal and postmortem adult brain cells, respectively). The data obtained support the hypothesis regarding aneuploidy occurrence in normal developing and adult human brain. PMID- 15750027 TI - The usefulness of calyculin a for cytogenetic prenatal diagnosis. AB - An increased number of chromosome plates can be obtained by use of calyculin A (CLA). CLA is an inhibitor of protein phosphatases (type 1 and type 2A serine/threonine). Inactivation of these phosphatases leads to premature chromosome condensation (PCC) in all phases of the cell cycle; thus, it is possible to investigate both metaphase and G(2)-PCC chromosomes. Amniotic fluid (AF) cultures were treated with calyculin A (CLA). GTG banding was obtained. Using this method it is possible to investigate all cell cycle phases, GTG banding, chromosomal breaks, and rates of PCD on the same preparation. Analyses of AF cultures treated with CLA allow complex studies on fetal genetic material. This work presents potential usefulness of CLA for cytogenetic prenatal diagnosis. PMID- 15750028 TI - The use of peptide nucleic acids for in situ identification of human chromosomes. AB - The peptide nucleic acids (PNAs) constitute a remarkable new class of synthetic nucleic acid analogues, based on their peptide-like backbone. This structure gives to PNAs the capacity to hybridize with high affinity and specificity to complementary RNA and DNA sequences and a great resistance to nucleases and proteinases. Originally conceived as ligands for the study of double-stranded DNA, the unique physicochemical properties of PNAs have led to the development of a large variety of research and diagnostic assays, including antigene and antisense therapy, genome mapping, and mutation detection. Over the past few years, PNAs have been shown to be powerful tools in cytogenetics for the rapid in situ identification of human chromosomes and the detection of aneuploidies. Recent studies have reported the successful use of chromosome-specific PNA probes on human lymphocytes, amniocytes, and spermatozoa, as well as on isolated oocytes and blastomeres. Multicolor PNA protocols have been described for the identification of several human chromosomes, indicating that PNAs could become a powerful complement to FISH for in situ chromosomal investigation. PMID- 15750029 TI - An approach for quantitative assessment of fluorescence in situ hybridization (FISH) signals for applied human molecular cytogenetics. AB - A number of applied molecular cytogenetic studies require the quantitative assessment of fluorescence in situ hybridization (FISH) signals (for example, interphase FISH analysis of aneuploidy by chromosome enumeration DNA probes; analysis of somatic pairing of homologous chromosomes in interphase nuclei; identification of chromosomal heteromorphism after FISH with satellite DNA probes for differentiation of parental origin of homologous chromosome, etc.). We have performed a pilot study to develop a simple technique for quantitative assessment of FISH signals by means of the digital capturing of microscopic images and the intensity measuring of hybridization signals using Scion Image software, commonly used for quantification of electrophoresis gels. We have tested this approach by quantitative analysis of FISH signals after application of chromosome-specific DNA probes for aneuploidy scoring in interphase nuclei in cells of different human tissues. This approach allowed us to exclude or confirm a low-level mosaic form of aneuploidy by quantification of FISH signals (for example, discrimination of pseudo-monosomy and artifact signals due to over-position of hybridization signals). Quantification of FISH signals was also used for analysis of somatic pairing of homologous chromosomes in nuclei of postmortem brain tissues after FISH with "classical" satellite DNA probes for chromosomes 1, 9, and 16. This approach has shown a relatively high efficiency for the quantitative registration of chromosomal heteromorphism due to variations of centromeric alphoid DNA in homologous parental chromosomes. We propose this approach to be efficient and to be considered as a useful tool in addition to visual FISH signal analysis for applied molecular cytogenetic studies. PMID- 15750030 TI - New aspects of laser microdissection in research and routine. AB - Laser microdissection has opened a window to new technologies. The scientific fields of genomics, transcriptomics, and proteomics need pure samples for rendering reliable results. Homogeneous sample preparation is a prerequisite for modern molecular analyses, both qualitative and quantitative. Laser microdissection and pressure catapulting (LMPC) is a tool for isolating specific cells from complex tissues in a non-contact and contamination-free manner. Because LMPC technology is an optimal method for obtaining fast and reliable access to single cells, the possibility of automatic isolation of single fetal cells has the promise of being a big step forward in developing protocols for non invasive prenatal diagnosis. PMID- 15750031 TI - Molecular karyotyping: array CGH quality criteria for constitutional genetic diagnosis. AB - Array CGH (comparative genomic hybridization) enables the identification of chromosomal copy number changes. The availability of clone sets covering the human genome opens the possibility for the widespread use of array CGH for both research and diagnostic purposes. In this manuscript we report on the parameters that were critical for successful implementation of the technology, assess quality criteria, and discuss the potential benefits and pitfalls of the technology for improved pre- and postnatal constitutional genetic diagnosis. We propose to name the genome-wide array CGH "molecular karyotyping," in analogy with conventional karyotyping that uses staining methods to visualize chromosomes. PMID- 15750032 TI - Origin, structure, and activity in vitro and in vivo of dalbavancin. AB - Dalbavancin is a novel semi-synthetic lipoglycopeptide that was designed to improve upon the natural glycopeptides currently available, vancomycin and teicoplanin. Chemical modification of natural glycopeptides has produced compounds with more potent antimicrobial activity and longer t(1/2), while maintaining an excellent safety profile. Dalbavancin, prepared from a teicoplanin like glycopeptide, has better activity, in vitro and in animal infection models, than vancomycin and teicoplanin. In particular, dalbavancin has excellent activity against staphylococci, including coagulase-negatives. A unique feature of dalbavancin is its pharmacokinetics, characterized by a long elimination t(1/2) in humans which makes a once-weekly dosing regimen feasible. Dalbavancin recently completed Phase 3 clinical trials in skin and skin structure infection. PMID- 15750033 TI - In vitro antistaphylococcal activity of dalbavancin, a novel glycopeptide. AB - OBJECTIVES: Dalbavancin is a novel, semi-synthetic glycopeptide antibiotic. The aim of this study was to further explore its activity against staphylococci. METHODS: The bactericidal activity of dalbavancin was studied using MBC and time kill methods. The potential for resistance to dalbavancin was examined using single-step and serial-passage experiments. RESULTS: Dalbavancin was bactericidal against methicillin-susceptible and -resistant Staphylococcus aureus, in both the presence and absence of human serum. No resistance was seen with any isolate tested. After serial passage, bacterial populations were more homogeneous in their susceptibility to dalbavancin than to vancomycin or teicoplanin. CONCLUSION: Dalbavancin is bactericidal for staphylococci. Resistance to this semi-synthetic glycopeptide is not readily developed in vitro. PMID- 15750034 TI - Human pharmacokinetics and rationale for once-weekly dosing of dalbavancin, a semi-synthetic glycopeptide. AB - The selection of a novel weekly dalbavancin dosage regimen was based on pharmacokinetic and pharmacodynamic data from humans and an animal model of infection. The results from the granuloma pouch model of infection suggested that dalbavancin concentrations >/=5 mg/L are necessary for extended in vivo activity. Serum bactericidal activity assessments demonstrated that dalbavancin serum concentrations of approximately 20 mg/L were bactericidal upon two-fold dilution. These data, coupled with simulations based on the pharmacokinetic profile derived from a clinical study in healthy volunteers, were used to design the weekly regimen studied in the initial efficacy trial. This efficacy study showed that a two-dose weekly regimen was well tolerated and associated with a higher clinical response rate than the comparator regimens. The data collectively support the further study of dalbavancin as a once-weekly regimen for the treatment of infections caused by Gram-positive bacteria. PMID- 15750035 TI - Pharmacokinetics and excretion of dalbavancin in the rat. AB - OBJECTIVES: The pharmacokinetics, tissue distribution and excretion routes of dalbavancin, a semi-synthetic glycopeptide, were investigated in rats. METHODS: A 20 mg/kg intravenous dose of dalbavancin or [(3)H]dalbavancin was administered to rats in three studies. Concentrations of dalbavancin or drug-derived radioactivity were assessed in blood, plasma, tissues, bile, urine and faeces by HPLC-MS/MS, scintillation counting or microbiological methods. RESULTS: Dalbavancin decayed tri-exponentially in plasma with an apparent terminal t(1/2) of 187 h (approximately 8 days). Dalbavancin has dual routes of elimination, with around two-thirds of the excreted drug-derived radioactivity being found in the urine and around one-third in the faeces. After 70 days, 44.2% and 22.3% of the drug-derived radioactivity had been recovered in the urine and faeces, respectively. Biliary excretion of drug-derived radioactivity accounted for over half of the radioactivity excreted faecally. At 70 days post-dose, <5% of the dose remained in the carcass, showing that drug elimination was complete. CONCLUSIONS: Dalbavancin has a long t(1/2) (approximately 8 days) in the rat and distributes widely throughout the body. It is not selectively retained in any single organ, tissue or blood component and is completely eliminated by both renal and non-renal routes in rats. These data were useful in designing and interpreting animal infection model studies used to select the dose for human studies. PMID- 15750036 TI - Safety and efficacy of glycopeptide antibiotics. AB - It would be difficult to envision the practice of infectious diseases over the past 20 years without the availability of the glycopeptide antibiotics. The two agents currently in clinical use, vancomycin and teicoplanin, have proven remarkably versatile in many common applications. Several attributes of these agents account for this favourable profile: (i) their broad spectrum of activity against Gram-positive bacteria, including strains resistant to many other antimicrobials; (ii) their favourable pharmacokinetic properties that allow the once- or twice-daily dosing regimens that have made out-of-hospital therapy possible; and (iii) their generally good safety profiles which, along with their structural dissimilarity to beta-lactam and other antimicrobials, permits their use in many patients who are intolerant of other antibiotic regimens. It is not entirely surprising, therefore, that despite more than 40 years of clinical use and the interim appearance of bacterial strains resistant to this drug class, there remains continued interest in the development of newer members of the glycopeptide antibiotic class. This paper is intended to provide a global overview of the efficacy and safety of glycopeptide antibiotics currently in use, as background to understanding the need for and potential roles of new agents of this class. PMID- 15750038 TI - Changes in upper airway size during tidal breathing in children with obstructive sleep apnea syndrome. AB - We performed respiratory-gated magnetic resonance imaging to evaluate airway dynamics during tidal breathing in 10 children with obstructive sleep apnea syndrome (OSAS; age, 4.3 +/- 2.3 years) and 10 matched control subjects (age, 5.0 +/- 2.0 years). We hypothesized that respiratory cycle fluctuations in upper airway cross-sectional area would be larger in children with OSAS. METHODS: Studies were performed under sedation. Respiratory gating was performed automatically at 10, 30, 50, 70, and 90% of inspiratory and expiratory volume. Airway cross-sectional area was measured at four ascending oropharyngeal levels at each increment of the respiratory cycle. RESULTS: We noted the following in subjects with OSAS compared with control subjects: (1) a smaller upper airway cross-sectional area, particularly during inspiration; (2) airway narrowing occurred during inspiration without evidence of complete airway collapse; (3) airway dilatation occurred during expiration, particularly early in the phase; and (4) magnitude of cross-sectional areas fluctuations during tidal breathing noted in OSAS at levels 1 through 4 were 317, 422, 785, and 922%, compared with 19, 15 17, and 24% in control subjects (p < 0.001, p < 0.005, p < 0.001, and p < 0.001, respectively). CONCLUSIONS: Fluctuations in airway area during tidal breathing are significantly greater in subjects with OSAS compared with control subjects. Resistive pressure loading is a probable explanation, although increased airway compliance may be a contributing factor. PMID- 15750039 TI - Mislocalization of DNAH5 and DNAH9 in respiratory cells from patients with primary ciliary dyskinesia. AB - RATIONALE: Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized by recurrent infections of the airways and situs inversus in half of the affected offspring. The most frequent genetic defects comprise recessive mutations of DNAH5 and DNAI1, which encode outer dynein arm (ODA) components. Diagnosis of PCD usually relies on electron microscopy, which is technically demanding and sometimes difficult to interpret. METHODS: Using specific antibodies, we determined the subcellular localization of the ODA heavy chains DNAH5 and DNAH9 in human respiratory epithelial and sperm cells of patients with PCD and control subjects by high-resolution immunofluorescence imaging. We also assessed cilia and sperm tail function by high-speed video microscopy. RESULTS: In normal ciliated airway epithelium, DNAH5 and DNAH9 show a specific regional distribution along the ciliary axoneme, indicating the existence of at least two distinct ODA types. DNAH5 was completely or only distally absent from the respiratory ciliary axoneme in patients with PCD with DNAH5- (n = 3) or DNAI1- (n = 1) mutations, respectively, and instead accumulated at the microtubule-organizing centers. In contrast to respiratory cilia, sperm tails from a patient with DNAH5 mutations had normal ODA heavy chain distribution, suggesting different modes of ODA generation in these cell types. Blinded investigation of a large cohort of patients with PCD and control subjects identified DNAH5 mislocalization in all patients diagnosed with ODA defects by electron microscopy (n = 16). Cilia with complete axonemal DNAH5 deficiency were immotile, whereas cilia with distal DNAH5 deficiency showed residual motility. CONCLUSIONS: Immunofluorescence staining can detect ODA defects, which will possibly aid PCD diagnosis. PMID- 15750040 TI - Inducible nitric oxide synthase in long-term intermittent hypoxia: hypersomnolence and brain injury. AB - RATIONALE: Long-term intermittent hypoxia (LTIH) exposure in adult mice, modeling oxygenation patterns of moderate-severe obstructive sleep apnea, results in lasting hypersomnolence and is associated with nitration and oxidation injuries in many brain regions, including wake-active regions. OBJECTIVES: We sought to determine if LTIH activates inducible nitric oxide synthase (iNOS) in sleep/wake regions, and if this source of NO contributes to the LTIH-induced proinflammatory gene response, oxidative injury, and wake impairments. METHODS: Mice with genetic absence of iNOS activity and wild-type control animals were exposed to 6 weeks of long-term hypoxia/reoxygenation before behavioral state recordings, molecular and biochemical assays, and a pharmacologic intervention. MEASUREMENTS AND MAIN RESULTS: Two weeks after recovery from hypoxia/reoxygenation exposures, wild-type mice showed increased iNOS activity in representative wake-active regions, increased sleep times, and shortened sleep latencies. Mutant mice, with higher baseline sleep times, showed no effect of long-term hypoxia/reoxygenation on sleep time latencies and were resistant to hypoxia/reoxygenation increases in lipid peroxidation and proinflammatory gene responses (tumor necrosis factor alpha and cyclooxygenase 2). Inhibition of iNOS after long-term hypoxia/reoxygenation in wild-type mice was effective in reversing the proinflammatory gene response. CONCLUSIONS: These data support a critical role for iNOS activity in the development of LTIH wake impairments, lipid peroxidation, and proinflammatory responses in wake-active brain regions, and suggest a potential role for inducible NO inhibition in protection from proinflammatory responses, oxidative injury, and residual hypersomnolence in obstructive sleep apnea. PMID- 15750041 TI - Activated protein C inhibits local coagulation after intrapulmonary delivery of endotoxin in humans. AB - RATIONALE: Acute lung injury and pneumonia are associated with pulmonary activation of coagulation and suppression of fibrinolysis, resulting in fibrin deposition in the lung. Activated protein C (APC) has systemic anticoagulant effects in patients with sepsis. OBJECTIVE: To determine the effect of systemic administration of recombinant human APC on endotoxin-induced hemostatic alterations in the bronchoalveolar space in humans. METHODS: Healthy humans received intravenous APC (24 microg/kg/hour; n = 8) or vehicle (n = 7); all subjects were administered saline in one lung subsegment and endotoxin (4 ng/kg) into the contralateral lung. Bronchoalveolar lavage was performed 16 hours after saline and endotoxin administration. MEASUREMENTS AND MAIN RESULTS: Endotoxin induced local activation of coagulation, as reflected by elevated levels of thrombin-antithrombin complexes (1.9 +/- 0.1 ng/ml) and soluble tissue factor (15.0 +/- 0.6 pg/ml) in bronchoalveolar lavage fluid, which was inhibited by APC (1.4 +/- 0.1 ng/ml and 12.3 +/- 0.4 pg/ml, respectively; both p < 0.01). Concurrently, endotoxin suppressed fibrinolysis, as indicated by reduced bronchoalveolar levels of plasminogen activator activity accompanied by elevated levels of plasminogen activator inhibitor type I activity. APC diminished the rise in plasminogen activator inhibitor type I activity (from 3.9 +/- 0.1 to 3.0 +/- 0.2 ng/ml, p = 0.002), while not significantly influencing plasminogen activator activity levels. Endotoxin reduced bronchoalveolar protein C concentrations, which was prevented by APC. Protein C did not influence the endotoxin-induced rise in local soluble thrombomodulin levels. CONCLUSION: APC exerts an anticoagulant effect in the human lung challenged with endotoxin. PMID- 15750042 TI - Sildenafil versus Endothelin Receptor Antagonist for Pulmonary Hypertension (SERAPH) study. AB - RATIONALE: Phosphodiesterase type 5 (PDE5) inhibition has been proposed for the treatment for pulmonary arterial hypertension (PAH). OBJECTIVE: This study compared adding sildenafil, a PDE5 inhibitor, to conventional treatment with the current practice of adding bosentan, an endothelin receptor antagonist. METHODS: Twenty-six patients with PAH, idiopathic or associated with connective tissue disease, World Health Organization (WHO) functional class III, were randomized in a double-blind fashion to receive sildenafil (50 mg twice daily for 4 weeks, then 50 mg three times daily) or bosentan (62.5 mg twice daily for 4 weeks, then 125 mg twice daily) over 16 weeks. MEASUREMENTS: Changes in right ventricular (RV) mass (using cardiovascular magnetic resonance), 6-minute walk distance, cardiac function, brain natriuretic peptide, and Borg dyspnea index. MAIN RESULTS: When analyzed by intention to treat, there were no significant differences between the two treatment groups. One patient on sildenafil died suddenly. Patients on sildenafil who completed the protocol showed significant changes from baseline, namely, reductions in RV mass (-8.8 g; 95% confidence interval [CI], -2, -16; n = 13, p = 0.015) and plasma brain natriuretic peptide levels (-19.4 fmol x ml(-1); 95% CI, -5, -34; p = 0.014) and improvements in 6-minute walk distance (114 m; 95% CI, 67, 160; p = 0.0002), cardiac index (0.3 L x min(-1) x m(-2); 95% CI, 0.1, 0.4; p = 0.008), and systolic left ventricular eccentricity index (-0.2; 95% CI, -0.02, -0.37; p = 0.031). Bosentan improved 6-minute walk distance (59 m; 95% CI, 29, 89; n = 12, p = 0.001) and cardiac index (0.3; 95% CI, 0.1, 0.4; p = 0.008). CONCLUSIONS: Sildenafil added to conventional treatment reduces RV mass and improves cardiac function and exercise capacity in patients with PAH, WHO functional class III. Safety monitoring is important until more experience is obtained. PMID- 15750043 TI - Atrial overdrive pacing in patients with sleep apnea with implanted pacemaker. AB - RATIONALE: Atrial overdrive pacing markedly improved sleep-disordered breathing in a recent study. OBJECTIVES: Using a single-blind, randomized, crossover design, we aimed to reproduce these findings and investigate the possible underlying mechanisms. METHODS: Twenty ambulatory patients with an implanted pacemaker or cardioverter defibrillator were studied by polysomnography on 3 consecutive nights in a randomized, single-blind, crossover study in which devices were programmed for nonpacing or for overdrive pacing at 7 or 15 beats/minute faster than the mean nocturnal heart rate. Ventilation and biomarkers (urinary norepinephrine excretion, amino-terminal portion of the precursor of brain natriuretic peptide, or NT-proBNP, were also evaluated. MEASUREMENTS AND MAIN RESULTS: Neither the primary endpoint apnea-hypopnea index, nor the apnea index, oxygen desaturation, ventilation, or biomarkers were affected by the nocturnal atrial overdrive pacing. A small, clinically insignificant, rate-dependent reduction in the hypopnea index was evoked by pacing (nonpacing, 13.4 +/- 1.4; pacing 7, 12.9 +/- 1.4; pacing 15, 10.9 +/- 1.0; p < 0.01, analysis of variance). CONCLUSIONS: The lack of effect on the apnea hypopnea index means that atrial overdrive pacing is inappropriate for treating sleep-disordered breathing. PMID- 15750044 TI - Detection of lung cancer by sensor array analyses of exhaled breath. AB - RATIONALE: Electronic noses are successfully used in commercial applications, including detection and analysis of volatile organic compounds in the food industry. OBJECTIVES: We hypothesized that the electronic nose could identify and discriminate between lung diseases, especially bronchogenic carcinoma. METHODS: In a discovery and training phase, exhaled breath of 14 individuals with bronchogenic carcinoma and 45 healthy control subjects or control subjects without cancer was analyzed. Principal components and canonic discriminant analysis of the sensor data was used to determine whether exhaled gases could discriminate between cancer and noncancer. Discrimination between classes was performed using Mahalanobis distance. Support vector machine analysis was used to create and apply a cancer prediction model prospectively in a separate group of 76 individuals, 14 with and 62 without cancer. MAIN RESULTS: Principal components and canonic discriminant analysis demonstrated discrimination between samples from patients with lung cancer and those from other groups. In the validation study, the electronic nose had 71.4% sensitivity and 91.9% specificity for detecting lung cancer; positive and negative predictive values were 66.6 and 93.4%, respectively. In this population with a lung cancer prevalence of 18%, positive and negative predictive values were 66.6 and 94.5%, respectively. CONCLUSION: The exhaled breath of patients with lung cancer has distinct characteristics that can be identified with an electronic nose. The results provide feasibility to the concept of using the electronic nose for managing and detecting lung cancer. PMID- 15750045 TI - Role of 15-deoxy delta(12,14) prostaglandin J2 and Nrf2 pathways in protection against acute lung injury. AB - RATIONALE: Acute lung injury (ALI) is a disease process that is characterized by diffuse inflammation in the lung parenchyma. Recent studies demonstrated that cyclooxygenase-2 (COX-2) induced at the late phase of inflammation aids in the resolution of inflammation by generating 15-deoxy-delta(12,14)-prostaglandin J2 (15d-PGJ2). Transcription factor Nrf2 is activated by electrophiles and exerts antiinflammatory effects by inducing the gene expression of antioxidant and detoxification enzymes. OBJECTIVES: Because 15d-PGJ2 is an endogenous electrophile, we hypothesized that it protects against ALI by activating Nrf2. METHODS: To test this hypothesis, we generated a reversible ALI model by intratracheal injection of carrageenin, an inducer of acute inflammation, whose stimulation has been known to induce COX-2. MAIN RESULTS: We found that ALI induced by carrageenin was markedly exacerbated in Nrf2-knockout mice, compared with wild-type mice. Analysis of bronchoalveolar lavage fluids also revealed that the magnitude and the duration of acute inflammation, indicated by albumin concentration and the number of neutrophils, were significantly enhanced in Nrf2 knockout mice. Treatment of wild-type mice with NS-398, a selective COX-2 inhibitor, significantly exacerbated ALI to the level of Nrf2-knockout mice. In the lungs of NS-398-treated wild-type mice, both the accumulation of 15d-PGJ2 and the induction of Nrf2 target antioxidant genes were significantly attenuated. Exogenous administration of 15d-PGJ2 reversed the exacerbating effects of NS-398 with the induction of antioxidant genes. CONCLUSIONS: These results demonstrated in vivo that 15d-PGJ2 plays a protective role against ALI by exploiting the Nrf2 mediated transcriptional pathway. PMID- 15750047 TI - Reproducibility of physical performance and physiologic assessments. AB - We evaluate the test-retest stability of physical performance and physiologic assessments used in epidemiologic research. METHOD: Eighty subjects aged 50 to 65 were randomly selected from a probability sample of African Americans for test retest assessments 5 to 45 days after baseline. Physical performance assessments included grip strength, chair stands, gait speed, and four standing-balance measures. Physiologic assessments included systolic and diastolic blood pressure, height, weight, body fat, and peak expiratory flow. RESULTS: Intraclass correlations coefficients (ICCs) were .81 for grip strength, .72 for chair stands, .56 for gait speed, .60 for one-leg stand, .52 for semitandem stand, .58 for tandem stand with eyes closed, and .27 for tandem stand with eyes open. Except for blood pressure (ICCs of .51 and .55 for systolic and diastolic), the physiologic assessments had ICCs > .89. DISCUSSION: Additional interviewer training may improve the reproducibility of the tandem stand with eyes open. PMID- 15750046 TI - Study of C-C chemokine receptor 2 alleles in sarcoidosis, with emphasis on family based analysis. AB - Prevailing models of sarcoidosis pathogenesis involve the activation of alveolar macrophages, aggregation of CD4+ T lymphocytes, and their accumulation in epithelioid cell granulomas. Increasing evidence suggests that each of these steps is modified by the host genetic constitution. Consequently, candidate susceptibility genes have been selected based on their potential function under this model. The C-C chemokine receptor 2 (CCR2) is involved in Th1 immune activity by recruiting competent cells and possibly by balancing response. CCR2 gene variants have been shown to be associated with sarcoidosis or, more specifically, with Lofgren's syndrome, a distinct form of acute sarcoidosis. We have studied three CCR2 gene polymorphisms (c.190G>A, c.840C>T, and c.4385A>T) in an extended sample of 1,203 patients with sarcoidosis and their relatives. Case control comparisons and family-based genetic analyses did not support previous findings of an association between CCR2 gene variability and the risk of sarcoidosis. However, they confirmed linkage disequilibrium and showed positive linkage results (p = 0.034) and therefore suggest a susceptibility gene in the surrounding chromosomal region. PMID- 15750048 TI - Caregiver burden and depressive symptoms: analysis of common outcomes in caregivers of elderly patients. AB - Despite widespread use of caregiver burden and depressive symptoms in caregiving research, the relationship between these two concepts and the way in which burden and depressive symptoms are affected are not clear. METHODS: The authors used structural equation modeling with an inception cohort of 488 family caregivers to examine the relationship between care recipients' mental and functional status and recency of care demands and caregivers' burden and depressive symptoms. RESULTS: Care recipients' mental and functional status and recency of care demands predicted caregiver burden; burden, in turn, was nearly significant in predicting depressive symptoms. Care recipients' mental status and recency of care demands had a near significant indirect effect on caregiver depressive symptoms. There were no significant direct paths between care recipients' mental status, functional status, recency of care demands, and caregivers' depressive symptoms. DISCUSSION: Health care practitioners should assist caregivers with new care demands stemming from care recipients' mental and functional status to decrease burdens and should monitor caregivers with higher levels of burden for the development of depressive symptoms. PMID- 15750049 TI - Caregiver appraisals of functional dependence in individuals with dementia and associated caregiver upset: psychometric properties of a new scale and response patterns by caregiver and care recipient characteristics. AB - To evaluate psychometric properties and response patterns of the Care-giver Assessment of Function and Upset (CAFU), a 15-item multidimensional measure of dependence in dementia patients and caregiver reaction. METHOD: 640 families were administered the CAFU (53% White, 43% African American, and 4% mixed race and ethnicity). We created a random split of the sample and conducted exploratory factor analyses on Sample 1 and confirmatory factor analyses on Sample 2. Convergent and discriminant validity were evaluated using Spearman rank correlation coefficients. RESULTS: A two-factor structure for functional items was derived, and excellent factorial validity was obtained. Convergent and discriminant validity were obtained for function and upset measures. Differential response patterns for dependence and caregiver upset were found for caregiver race, relationship, and care recipient gender but not for caregiver gender. DISCUSSION: The CAFU is easily administered, reliable, and valid for evaluating appraisals of dependencies and upsetting care areas. PMID- 15750050 TI - Caregiver psychological adjustment and institutionalization of persons with Alzheimer's disease. AB - This prospective study examines the relationship between caregiver psychological adjustment and institutionalization of persons with Alzheimer's disease (AD). METHOD: A 3-year longitudinal study was conducted with a clinic-based cohort of 396 persons with AD and their respective family caregivers. Caregiver adjustment was sequentially measured in 3-month intervals using standard scales of appraisal (perceived burden and satisfaction) and emotion (depressive symptoms and positive affect). RESULTS: In multivariable proportional hazards regression models predicting time to institutionalization controlling for AD severity, hazard ratios were significant for appraisal measures of caregiver adjustment, perceived burden (1.053; 95% confidence interval [CI], 1.014, 1.093), and satisfaction (.929; 95% CI, .883, .977). In contrast, levels of caregivers' positive and negative emotion did not reliably predict institutionalization. DISCUSSION: The findings highlight the importance of caregiver appraisals in decisions to institutionalize persons with a dementia syndrome, but the limited impact of caregiver emotion was unexpected and requires further study. PMID- 15750051 TI - Factors affecting choice of community residential care setting. AB - To investigate factors associated with residents' choice of type of Medicaid funded community residential care setting in western Washington State. METHOD: Prospective cohort design including residents new to any of three setting types (264 residents entering 170 different facilities), using data from state and Medicaid databases and in-person interviews. The authors used analysis of variance and multinomial logistic regression to examine bivariate associations and estimate effects of resident and facility characteristics on choice of facility type at baseline. RESULTS: Several resident characteristics appear to be associated with choice of community residential care setting, including age, marital status, education, functional status, and reported memory and behavior problems. Facility policies differ significantly among types of facilities and also appear to be associated with choice of setting. DISCUSSION: Selection processes operate in choice of community residential care setting, with residents choosing facility type based on the fit of their needs with facility characteristics. PMID- 15750052 TI - Discretionary hospitalization of nursing home residents with and without Alzheimer's disease: a multilevel analysis. AB - This study analyzes facility variations in hospital admission rates of nursing home (NH) residents with and without Alzheimer's disease (AD) or related dementia with the aim of better understanding how facility-level contextual factors differentially affect hospitalization risks. METHOD: The sample population consists of 19,217 and 18,399 Medicaid residents with and without AD, respectively, from 546 NHs in Massachusetts between 1991 and 1993. Hospital use is measured as annual nonpsychiatric discretionary hospital admissions to short term general hospitals. Multilevel estimation methods are used to obtain facility and market area parameter estimates. RESULTS: There was greater interfacility variation in discretionary hospital admission rates of AD residents than residents without AD, particularly among more vulnerable subgroups of AD residents. DISCUSSION: The findings underscore the importance of licensed nursing personnel in reducing discretionary hospitalizations among NH residents with AD. PMID- 15750053 TI - Modifiable impairments predict progressive disability among older persons. AB - Our purpose was to determine the extent to which a predetermined set of modifiable impairments predicted progression of disability. METHOD: We conducted a 3-year follow-up of two community-based cohorts of older adults. The impairment areas included lower extremity, upper extremity, hearing, vision, and affect. Home management and social or productive activities were the domains of function investigated. RESULTS: All five impairments were of at least borderline significance in predicting decline in both functional domains in both cohorts with the exception of hearing for home management activities. The five impairments together explained from 17% to 23% of the decline seen in the functional outcomes (partial R(2)s 0.17 to 0.23). DISCUSSION: Five prevalent and potentially modifiable impairments explained much of the progressive disability experienced. Given the priority that older patients place on function as a health outcome, these impairments should be routinely assessed and modified. PMID- 15750054 TI - Detection of cryptosporidium and identification to the species level by nested PCR and restriction fragment length polymorphism. AB - Cryptosporidiosis is an emerging protozoan disease associated with large waterborne outbreaks. Diagnosis relies on microscopic examination of stools, but this method cannot identify the infecting species of Cryptosporidium. We have developed a test based on nested PCR and restriction fragment length polymorphism (RFLP) that offers simple identification of Cryptosporidium hominis, Cryptosporidium parvum, and most other human infective species in stool samples. Purified C. parvum oocysts were used for PCR development. Extracted DNA was amplified by nested PCR targeting a 214-bp fragment of the 18S RNA gene. Enzymatic restriction sites were identified by bioinformatic analysis of all published Cryptosporidium 18S rRNA sequences. Experiments with spiked stool samples gave an estimated PCR detection limit of one oocyst. Specificity was assessed by testing 68 stool samples from patients with microscopically proven cryptosporidiosis and 31 Cryptosporidium-negative stools. Sixty-seven (98.5%) of the 68 stool samples from patients with microscopically proven cryptosporidiosis and 2 of the other stool samples were positive by PCR and could be genotyped. RFLP analysis identified 36 C. hominis, 19 C. parvum, 8 Cryptosporidium meleagridis, and 6 Cryptosporidium felis or Cryptosporidium canis samples. Species determination in 26 PCR-positive cases was in full agreement with DNA sequencing of the 18S rRNA hypervariable region. The excellent sensitivity of PCR, coupled with the accuracy of RFLP for species identification, make this method a suitable tool for routine diagnosis and genotyping of Cryptosporidium in stools. PMID- 15750055 TI - Use of diagnostic microarrays for determination of virulence gene patterns of Escherichia coli K1, a major cause of neonatal meningitis. AB - Forty Escherichia coli strains isolated primarily from neonatal meningitis, urinary tract infections and feces were screened for the presence of virulence genes with a newly developed microarray on the array tube format. A total of 32 gene probes specific for extraintestinal as well as intestinal E. coli pathotypes were included. Eighty-eight percent of the analyzed strains were positive for the K1-specific probe on the microarray and could be confirmed with a specific antiserum against the K1 capsular polysaccharide. The gene for the hemin receptor ChuA was predominantly found in 95% of strains. Other virulence genes associated with K1 and related strains were P, S, and F1C fimbriae specific for extraintestinal E. coli, the genes for aerobactin, the alpha-hemolysin and the cytotoxic necrotizing factor. In two strains, the O157-specific catalase gene and the gene for the low-molecular-weight heat-stable toxin AstA were detected, respectively. A total of 19 different virulence gene patterns were observed. No correlation was observed between specific virulence gene patterns and a clinical outcome. The data indicate that virulence genes typical of extraintestinal E. coli are predominantly present in K1 strains. Nevertheless, some of them can carry virulence genes known to be characteristic of intestinal E. coli. The distribution and combination of virulence genes show that K1 isolates constitute a heterogeneous group of E. coli. PMID- 15750056 TI - First description of Curtobacterium spp. isolated from human clinical specimens. AB - During a 4-year period, five strains (three of which were doubtless clinically significant) of yellow- or orange-pigmented, oxidative, slowly acid-producing coryneform bacteria were recovered from human clinical specimens in two reference laboratories or referred to them. The strains were motile, catalase positive, nitrate reductase negative, and urease negative, but strongly hydrolyzed esculin. In all reference and clinical strains described in the present study, anteisopentadecanoic (C(15:0ai)) and anteisoheptadecanoic (C(17:0ai)) acids represented more than 75% of all cellular fatty acids except in one clinical strain and in Curtobacterium pusillum, in which both the unusual omega-cyclohexyl fatty acid (identified as C(18:1omega7cis/omega9cis/omega12trans) by the Sherlock system) represented more than 50% of all cellular fatty acids. In all clinical strains, ornithine was the diamino acid of the cell wall, the interpeptide bridge consisted of ornithine, and acetyl was the acyl type of the peptidoglycan. Therefore, the five clinical strains were unambiguously identified as Curtobacterium spp. Analyses of the complete 16S rRNA genes of the five clinical strains with homologies to the established Curtobacterium species ranging from 99.2 to 100% confirmed the identifications as Curtobacterium spp. Data on the antimicrobial susceptibility pattern of curtobacteria are reported, with macrolides and rifampin showing very low MICs for all strains tested. This report is the first on the isolation of Curtobacterium strains from human clinical specimens. PMID- 15750058 TI - Establishment of a universal size standard strain for use with the PulseNet standardized pulsed-field gel electrophoresis protocols: converting the national databases to the new size standard. AB - The PulseNet National Database, established by the Centers for Disease Control and Prevention in 1996, consists of pulsed-field gel electrophoresis (PFGE) patterns obtained from isolates of food-borne pathogens (currently Escherichia coli O157:H7, Salmonella, Shigella, and Listeria) and textual information about the isolates. Electronic images and accompanying text are submitted from over 60 U.S. public health and food regulatory agency laboratories. The PFGE patterns are generated according to highly standardized PFGE protocols. Normalization and accurate comparison of gel images require the use of a well-characterized size standard in at least three lanes of each gel. Originally, a well-characterized strain of each organism was chosen as the reference standard for that particular database. The increasing number of databases, difficulty in identifying an organism-specific standard for each database, the increased range of band sizes generated by the use of additional restriction endonucleases, and the maintenance of many different organism-specific strains encouraged us to search for a more versatile and universal DNA size marker. A Salmonella serotype Braenderup strain (H9812) was chosen as the universal size standard. This strain was subjected to rigorous testing in our laboratories to ensure that it met the desired criteria, including coverage of a wide range of DNA fragment sizes, even distribution of bands, and stability of the PFGE pattern. The strategy used to convert and compare data generated by the new and old reference standards is described. PMID- 15750057 TI - Nosocomial outbreak caused by Salmonella enterica serotype Livingstone producing CTX-M-27 extended-spectrum beta-lactamase in a neonatal unit in Sousse, Tunisia. AB - In this study, we report an outbreak of Salmonella enterica serotype Livingstone resistant to extended-spectrum cephalosporins that occurred in a neonatal ward of the maternity department of Farhat Hached Hospital, Sousse, Tunisia, in 2002. A total of 16 isolates were recovered from 16 babies hospitalized in the ward during the period 1 to 16 July. All these babies developed diarrhea, and three of them developed septicemia. All the isolates demonstrated resistance to ceftriaxone and ceftazidime due to the production of an extended-spectrum beta lactamase (ESBL). The isolates were also resistant to aminoglycosides (kanamycin, tobramycin, netilmicin, gentamicin, and amikacin) and sulfamethoxazole trimethoprim. DNA profiles were determined by pulsed-field gel electrophoresis using the XbaI and SpeI endonucleases and by ribotyping with PstI digestion. They yielded the same patterns, showing that the outbreak was caused by a single clone. The ESBL was identified as CTX-M-27 by sequencing of PCR products and by isoelectric focusing. The ESBL resistance was transferred by a 40-kb conjugative plasmid. The mobile insertion sequence ISEcp1 was found to be located upstream of bla(CTX-M-27) in the same position as that known for a bla(CTX-M-14) sequence. A new gene named dfrA21, encoding resistance to trimethoprim and carried by a 90-kb plasmid, was characterized. The dfrA21 gene was inserted as a single resistance cassette in a class I integron. The babies were treated with colistin, and all except two recovered. The outbreak came to an end when appropriate actions were taken: patient isolation, hand washing, and disinfection of the ward. PMID- 15750060 TI - Comparison of specimen processing and nucleic acid extraction by the swab extraction tube system versus the MagNA Pure LC system for laboratory diagnosis of herpes simplex virus infections by LightCycler PCR. AB - A total of 563 specimens (234 dermal and 329 genital swabs) from patients suspected of having herpes simplex virus (HSV) infections were processed using two different extraction methods (the MagNA Pure LC system and the swab extraction tube system [SETS]); HSV DNA was amplified by LightCycler PCR. HSV DNA was detected in 157 of 563 specimens (27.9%) processed by the MagNA Pure LC system and in 179 of 563 specimens (31.8%) processed by SETS (P < 0.0001). There was no specimen processed by the MagNA Pure LC extraction method that was positive only for HSV DNA. Of 157 specimens positive by both methods, HSV DNA copy levels were higher (using cycle crossover points [cycle threshold {C(T)}]) with SETS (mean C(T), 25.9 cycles) than with the MagNA Pure LC system (mean C(T), 32.0 cycles) (P < 0.0001). The time to process 32 samples was longer with the MagNA Pure LC extraction system (90 min) than with SETS (35 min). HSV DNA extraction using SETS is faster, less expensive, and more sensitive than the MagNA Pure LC system and could replace the latter for the laboratory diagnosis of HSV infections using LightCycler PCR. PMID- 15750059 TI - Identification of Mycobacterium species by secA1 sequences. AB - We describe a novel molecular method for the differentiation and identification of 29 mycobacterial species. The target is the secA1 gene that codes for the essential protein SecA1, a key component of the major pathway of protein secretion across the cytoplasmic membrane. A 700-bp region of the secA1 gene was amplified and sequenced from 47 American Type Culture Collection strains of 29 Mycobacterium species as well as from 59 clinical isolates. Sequence variability in the amplified segment of the secA1 gene allowed the differentiation of all species except for the members of the Mycobacterium tuberculosis (MTB) complex, which had identical sequences. A range of 83.3 to 100% interspecies similarity was observed. All species could also be differentiated by their amino acid sequences as deduced from the sequenced region of the secA1 gene, with the exception of the MTB complex. Partial sequences of secA1 from clinical isolates belonging to nine frequently isolated species of mycobacteria revealed a very high intraspecies similarity at the DNA level (typically >99%; range, 96.0 to 100%); all clinical isolates were correctly identified. Comparison of the deduced 233-amino-acid sequences among clinical isolates of the same species showed between 99.6 and 100% similarity. To our knowledge, this is the first time a secretion-related gene has been used for the identification of the species within a bacterial genus. PMID- 15750061 TI - Multiplex PCR assay for rapid identification of oculopathogenic adenoviruses by amplification of the fiber and hexon genes. AB - Eye infections caused by adenovirus (Ad) often result in nosocomial infections and community epidemics with significant rates of morbidity. No antiviral agent effective against Ad is yet available for clinical use. Therefore, early diagnosis is still the mainstay for patient management and the prevention of epidemics. A multiplex PCR assay based on amplification of a combination of the fiber and hexon genes which can identify the six important oculopathogenic serotypes of Ads (Ad serotype 3[Ad3], Ad4, Ad7, Ad8, Ad19, and Ad37) in a single tube amplification reaction was developed. Ad serotypes could be distinguished by the different amplicon sizes. The assay correctly identified prototype strains as well as isolates in clinical specimens. In comparison with a previously described PCR-restriction fragment polymorphism method, our assay gave unequivocal results for clinical specimens. Our multiplex PCR has the potential to serve as a rapid and cost-effective tool for the typing of important ocular Ads. PMID- 15750062 TI - Rapid identification of the coxsackievirus A24 variant by molecular serotyping in an outbreak of acute hemorrhagic conjunctivitis. AB - We evaluated the clinical applicability of a molecular serotyping method for determination of the cause of epidemic acute hemorrhagic conjunctivitis. Seventy conjunctival swab specimens from individuals involved in a nationwide acute hemorrhagic conjunctivitis outbreak were tested. Viral culture and a molecular biology-based assay were compared by directly using clinical specimens. On the one hand, virus culture was done to isolate the enteroviruses, and serotyping was done by a coxsackievirus A24 variant-specific PCR. On the other hand, the original clinical specimens were directly screened for enterovirus by reverse transcription (RT)-PCR with panenterovirus-specific primers. Enterovirus screening-positive specimens were subjected to RT-PCR for detection of the VP1 region of enterovirus, and the amplicons were sequenced. Molecular serotyping was done by calculating the pairwise identity scores for the sequences with the maximum identities to the sequences of known prototype enteroviruses. Thirty-two specimens (45.7%) were culture positive, whereas 37 specimens (52.8%) were screening PCR positive (P < 0.001). The VP1 regions were amplified from 21 of the 37 specimens (56.8%), and the products amplified from 9 specimens were appropriately sequenced. These nine sequences were homologous with the sequence of the coxsackievirus A24 variant. Molecular serotyping by direct use of clinical specimens without cell culture could be applied for the rapid identification of the causative agent of epidemic acute hemorrhagic conjunctivitis. PMID- 15750064 TI - Sensitive and specific detection of staphylococcal epidermolysins A and B in broth cultures by flow cytometry-assisted multiplex immunoassay. AB - Two of the most common bacterial skin infections of young infants and children are bullous impetigo due to Staphylococcus aureus and its more acute form, staphylococcal scalded skin syndrome. Epidermolysin A (ETA), ETB and, possibly, ETD are responsible for these diseases, which may appear as epidemics in pediatric patients. We tested the reliability of a flow cytometry-assisted multiplex immunoassay (Bio-Plex system) for the detection of ETA and ETB. The Bio Plex system was found to be highly specific and highly sensitive for toxin concentrations of between 2 and 80,000 pg/ml. The results of this assay were 100% identical to the results of a PCR-based method. We demonstrated that this test did not generate any cross-reactions with ETD-producing isolates. The level of detection of ETB by this test differed according to culture conditions and from isolate to isolate; these results must be taken into account for diagnostic purposes. PMID- 15750063 TI - Extended-spectrum beta-lactamase-producing Shiga toxin gene (Stx1)-positive Escherichia coli O26:H11: a new concern. AB - Escherichia coli strain TUM2139 was isolated from a stool sample from a 9-year old girl on 16 June 2004. This strain was categorized as Shiga toxin-producing Escherichia coli (STEC) because the Shiga-like toxin gene stx(1) was detected by immunochromatography and PCR assay. The strain was highly resistant to cefotaxime (256 microg/ml) and was also resistant to cefepime, cefpodoxime, ceftriaxone, and aztreonam. In the presence of 4 microg of clavulanic acid per ml, the MIC of cefotaxime decreased to < or =0.12 microg/ml, indicating that this strain was an extended-spectrum beta-lactamase (ESBL) producer. Cefotaxime resistance was transferred to E. coli C600 by conjugation at a frequency of 3.0 x 10(-6). A PCR assay was performed with primer sets specific for TEM-type and SHV-type ESBLs and for the CTX-M-2 (Toho-1), CTX-M-3, and CTX-M-9 groups of ESBLs. A specific signal was observed with the primer set specific for the CTX-M-9 group of beta lactamases. This beta-lactamase was confirmed to be the ESBL CTX-M-18 by DNA sequencing. This is the first report of an ESBL-producing STEC isolate. PMID- 15750065 TI - Evaluation of four commercially available extended-spectrum beta-lactamase phenotypic confirmation tests. AB - Extended-spectrum beta-lactamase (ESBL) production in members of the Enterobacteriaceae can confer resistance to extended-spectrum cephalosporins, aztreonam, and penicillin. As such, the accurate detection of ESBL producers is essential for the appropriate selection of antibiotic therapy. Twenty previously characterized isolates and 49 clinical isolates suspected of ESBL production were tested by four ESBL phenotypic confirmatory methods for accuracy and ease of use. The four ESBL phenotypic confirmation tests included Dried MicroScan ESBL plus ESBL Confirmation panels (Dade Behring, Inc., West Sacramento, Calif.), Etest ESBL (AB BIODISK, Piscataway, N.J.), Vitek GNS-120 (bioMerieux, Inc., Hazelwood, Mo.), and BD BBL Sensi-Disk ESBL Confirmatory Test disks (BD Biosciences, Sparks, Md.). Results were compared to frozen microdilution panels prepared according to NCCLS specifications, and discrepant isolates were sent for molecular testing. The test sensitivities for the ESBL phenotypic confirmatory test methods used in this study were as follows: MicroScan ESBL plus ESBL confirmation panel, 100%; VITEK 1 GNS-120, 99%; Etest ESBL, 97%; and BD BBL Sensi-Disk ESBL Confirmatory Test disks, 96%. The test specificities were as follows: BD BBL Sensi-Disk ESBL Confirmatory Test disks, 100%; MicroScan ESBL plus ESBL confirmation panel and VITEK 1 GNS-120, 98%; and Etest ESBL, 94%. All methods were easy to perform; however, the Etest method required more expertise to interpret the results. All tests offer a feasible solution for confirming ESBL production in the clinical laboratory. PMID- 15750067 TI - Multiprefectural spread of gastroenteritis outbreaks attributable to a single genogroup II norovirus strain from a tourist restaurant in Nagasaki, Japan. AB - A series of gastroenteritis outbreaks caused by noroviruses (NVs) among tourist groups from several prefectures was associated with eating a lunch prepared by a restaurant in Nagasaki City, Japan, on 18 and 19 November 2003. A retrospective cohort study was performed to estimate the magnitude of the outbreak and identify the source of infection. Epidemiological information was obtained through the local public health centers in the areas where the illness occurred. Stool and vomit specimens and food and environmental samples were analyzed by reverse transcription-PCR with genogroup-specific primers. Positive samples were sequenced and analyzed phylogenetically. Of 1,492 tourists who ate a lunch prepared by the restaurant during the 2-day period, 660 (44.2%) developed illness, with an average incubation time of 31.2 h. Whereas NVs were not detected in any food samples, identical sequences most closely related to the Mexico genotype of genogroup II NV were found in specimens from case patients, restaurant staff, and the kitchen table. Food handlers were concluded to be the source of the outbreak as a result of the contamination of several meals. The series of outbreaks described here exemplifies the role of tourism as a contemporary way to distribute a single infectious agent to multiple and geographically remote areas. PMID- 15750066 TI - Molecular epidemiology of norovirus infections in Stockholm, Sweden, during the years 2000 to 2003: association of the GGIIb genetic cluster with infection in children. AB - The incidence of norovirus-associated gastroenteritis and the molecular epidemiology of norovirus strains were studied during three seasons (2000-2001, 2001-2002, and 2002-2003) among patients of all ages, mainly from the Stockholm region in Sweden. A total of 3,252 fecal samples were analyzed by reverse transcription-PCR. The incidences of norovirus infection among adults were 23, 26, and 30% during the three seasons studied and 18, 11, and 15% among children 0 to 15 years of age. During the first season, all norovirus strains detected by PCR were typed either by reverse line blot hybridization or nucleotide sequence analysis. During the two successive seasons, a total of 60 norovirus-positive strains from the beginning, peak, and end of the seasons were selected for nucleotide sequence analysis. We identified two dominant norovirus variants over the seasons: a new norovirus variant, recently described as the GGIIb genetic cluster, dominated among children during the first season, and during the following two seasons, a GGII-4 variant dominated. Our data suggest that norovirus infections are common, not only among adults, but also among children, and that some strains may predominantly affect children. PMID- 15750068 TI - Characterization of rotavirus strains in a Danish population: high frequency of mixed infections and diversity within the VP4 gene of P[8] strains. AB - We characterized the G and P types from 162 rotavirus-positive stool specimens collected from 162 persons in Denmark (134 children and 28 adults) with acute diarrhea in 1998, 2000, and 2002. Samples were obtained during outpatient consultations (73%) and from hospitalized patients (27%). Although more than 20 different G-P combinations were identified, only 52% represented the globally most common types G1P[8], G2P[4], and G4P[8]. The G9 genotype, which is emerging worldwide, was identified in 12% of all samples. Twenty-one percent of the samples were of mixed genotypic origin, which is the highest frequency reported in any European population. The standard reverse transcription-PCR methods initially failed to identify a considerable fraction of the rotavirus P strains due to mutations at the VP4 primer-binding sites of P[8] strains. The application of a degenerate P[8] primer resulted in typing of most VP4 strains. There was considerable year-to-year variation among the circulating G-P types, and whereas G1P[8] was predominant in 1998 (42% of samples) and 2002 (26%), G2P[4] was the strain that was most frequently detected in 2000 (26% of samples). Our findings might implicate challenges for rotavirus vaccine implementation in a European population and underscore the importance of extensive strain surveillance prior to, during, and after introduction of any vaccine candidate. PMID- 15750069 TI - Role of real-time molecular typing in the surveillance of Campylobacter enteritis and comparison of pulsed-field gel electrophoresis profiles from chicken and human isolates. AB - The goal of the present study was to assess the contribution of real-time molecular typing, used alone or with clinical surveillance, to the prompt identification of clusters of Campylobacter enteritis. Potential poultry sources were sought by comparing the pulsed-field gel electrophoresis genotypes of human and fresh whole retail chicken isolates collected during the same study period. Among 183 human isolates, 82 (45%) had unique genotypes, 72 (39%) represented 26 clusters of 2 to 7 isolates each, and 29 (16%) represented three clusters of 8 to 11 isolates each. Molecular typing was useful for the confirmation of outbreaks suspected on the basis of epidemiological surveillance, but for most small clusters, no epidemiological link could be established. Thus, the added value of real-time molecular typing is questionable, since the numerous small clusters identified were of unclear public health significance. Among 177 chickens, 41 (23%) yielded campylobacter isolates; of these, 19 (46%) had genotypes similar to those of 41 (22%) human isolates. However, a temporal association was demonstrated in only a minority of cases, and most genotypes were present only in a single species, suggesting that sources other than chickens are important in human campylobacteriosis. Further investigation with samples from water and other possible environmental sources is needed to define the most efficient strategy for the application of molecular typing and identification of the source(s) of sporadic cases of campylobacteriosis. PMID- 15750070 TI - Prevalence of norovirus among visitors from the United States to Mexico and Guatemala who experience traveler's diarrhea. AB - Traveler's diarrhea (TD) is the most common infectious illness acquired by visitors to developing nations. The purpose of this study was to utilize molecular diagnostic techniques to determine the prevalence of norovirus (NoV) in TD occurring among visitors from the United States to Guatemala and Mexico. Stool samples (n = 54) were collected from 34 TD cases and analyzed for NoV by reverse transcription-PCR and oligoprobe confirmation. The overall prevalence of NoV was 65%. Interestingly, all NoV-positive stool samples were identified as genogroup I NoVs, and time spent at travel destinations was found to be an important factor in determining the frequency of infection (P = 0.003). Eleven NoV-positive stool samples also tested positive for enterotoxigenic Escherichia coli, indicating that dual infections with this leading bacterial cause of TD were very common. Results of this study suggest that NoV infection is a frequent occurrence among travelers to Mexico and Guatemala who experience episodes of TD. In addition, the simple molecular detection method utilized here will serve to facilitate more in depth epidemiological studies of this emergent viral pathogen in travelers and other at-risk populations. PMID- 15750071 TI - Novel antibody-lectin enzyme-linked immunosorbent assay that distinguishes prion proteins in sporadic and variant cases of Creutzfeldt-Jakob disease. AB - We used different anti-prion protein (anti-PrP) monoclonal antibodies to capture either full-length or truncated PrP species and then used biotinylated lectin to compare the nature of the glycans on bound PrP species present in control, sporadic Creutzfeldt-Jakob disease (sCJD), or variant CJD (vCJD) brains. When full-length PrP species in these three groups were compared, no significant difference in the binding of concanavalin A or Aleuria aurantia lectin was detected. However, the binding of Ricinus communis agglutinin I (RCA) to sCJD and vCJD samples was significantly increased. In contrast, when only truncated PrP species were compared, only vCJD samples had more RCA binding activity. Therefore, while most of the RCA binding activity in sCJD is restricted to the full-length PrP species, the RCA binding activity in vCJD is associated with truncated and full-length PrP species. Furthermore, the RCA binding activity in sCJD and vCJD samples is mostly associated with proteinase K-resistant PrP species, a known signature of infectious prion. Therefore, PrP species in sCJD and vCJD have dissimilar lectin immunoreactivity, which reflects differences in their N-linked glycans. These differences may account for the distinct phenotypes of sCJD and vCJD. PMID- 15750072 TI - Detection and identification of Ehrlichia spp. in ticks collected in Tunisia and Morocco. AB - A broad-range 16S rRNA gene PCR assay followed by partial sequencing of the 16S rRNA gene was used for the detection of members of the family Anaplasmataceae in ticks in North Africa. A total of 418 questing Ixodes ricinus ticks collected in Tunisia and Morocco, as well as 188 Rhipicephalus ticks from dogs and 52 Hyalomma ticks from bovines in Tunisia, were included in this study. Of 324 adult I. ricinus ticks, 16.3% were positive for Ehrlichia spp., whereas only 3.4 and 2.8% of nymphs and larvae, respectively, were positive. A large heterogeneity was observed in the nucleotide sequences. Partial sequences identical to that of the agent of human granulocytic ehrlichiosis (HGE) were detected in I. ricinus and Hyalomma detritum, whereas partial sequences identical to that of Anaplasma platys were detected in Rhipicephalus sanguineus. However, variants of Anaplasma, provisionally designated Anaplasma-like, were predominant in the I. ricinus tick population in Maghreb. Otherwise, two variants of the genus Ehrlichia were detected in I. ricinus and H. detritum. Surprisingly, a variant of Wolbachia pipientis was evidenced from I. ricinus in Morocco. These results emphasized the potential risk of tick bites for human and animal populations in North Africa. PMID- 15750073 TI - Typing of Saccharomyces cerevisiae clinical strains by using microsatellite sequence polymorphism. AB - It seems that S. cerevisiae, which was thought for about 30 years to be a nonpathogenic yeast, should now be considered an opportunistic pathogen. In this study, we estimated the discrimination ability of the microsatellite sequence amplification technique within a sample of clinical and reference S. cerevisiae strains and S. boulardii reference strains. PMID- 15750074 TI - Detection of human metapneumovirus antigens in nasopharyngeal secretions by an immunofluorescent-antibody test. AB - Human metapneumovirus (hMPV) is a recently discovered pathogen associated with respiratory tract infections, primarily in young children, immunocompromised individuals, and elderly individuals. Reverse transcription-PCR (RT-PCR) has been reported to be a more sensitive method for the diagnosis of hMPV infections than virus isolation by culture and serological study. However, there has been no report on rapid methods, such as an immunofluorescent-antibody test or an enzyme linked immunosorbent assay, for the detection of hMPV antigens in nasopharyngeal secretions. In this study, we compared an indirect immunofluorescent-antibody test (IFA) with a monoclonal antibody with RT-PCR for detection of hMPV in nasal secretions from 48 hospitalized children with respiratory tract infections. Fifteen of the 48 children were positive for hMPV by RT-PCR. IFA results were positive for 11 of the 15 RT-PCR-positive children (sensitivity, 73.3%) and 1 of the 33 RT-PCR-negative children (specificity, 97.0%). Although the sensitivity of IFA is lower than that of RT-PCR, IFA is a rapid and useful test for the diagnosis of hMPV infections in children. PMID- 15750075 TI - Ascariasis is a zoonosis in denmark. AB - A preliminary epidemiological survey indicated an association between Ascaris infections in Danish patients and contact with pigs or pig manure. In the present study, we compared Ascaris worms collected from humans and Ascaris worms collected from pigs by amplified fragment length polymorphism (AFLP) analysis, a technique for whole-genome fingerprinting, and by PCR-linked restricted fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer region of nuclear rDNA. The AFLP data were analyzed by distance- and model-based clustering methods. These results assigned Ascaris worms from Danish patients to a cluster different from that for worms from humans in other geographic areas. In contrast, worms from humans and pigs in Denmark were assigned to the same cluster. These results were supported by the PCR-RFLP results. Thus, all of the examined Danish patients had acquired Ascaris infections from domestic pigs; ascariasis may therefore be considered a zoonotic disease in Denmark. PMID- 15750076 TI - Rapid identification of staphylococcal strains from positive-testing blood culture bottles by internal transcribed spacer PCR followed by microchip gel electrophoresis. AB - PCR analysis of the 16S-23S rRNA gene internal transcribed spacer (ITS) followed by microchip gel electrophoresis (MGE) was evaluated for its usefulness in identification of staphylococci. Forty ITS PCR patterns were demonstrated among 228 isolated colonies of Staphylococcus aureus: 26 patterns for methicillin susceptible S. aureus (MSSA; 91 strains), 11 patterns for methicillin-resistant S. aureus (MRSA; 99 strains), and 3 patterns for both MSSA and MRSA (38 strains). Thirty-seven control strains of coagulase-negative staphylococci (CNS) representing 16 species showed unique ITS PCR patterns (24 patterns) at the species and subspecies levels: two patterns for S. caprae, S. cohnii, S. haemolyticus, and S. saprophyticus; three patterns for S. lugdunensis; four patterns for S. capitis; and one pattern for each of the other CNS species. The combined PCR-MGE method was prospectively adapted to the positive blood culture bottles, and this method correctly identified MSSA and MRSA in 102 (89%) of 114 blood cultures positive for S. aureus on the basis of the ITS PCR patterns. Eight ITS PCR patterns were demonstrated from 166 blood culture bottles positive for CNS. The most frequent CNS species isolated from blood cultures were S. epidermidis (76%), S. capitis (11%), and S. hominis (8%). Overall, all 280 blood culture bottles shown to contain a single Staphylococcus species by routine phenotypic methods were correctly identified by the PCR-MGE method at the species level, whereas the organism failed to be identified in 8 culture bottles (3%) with mixed flora. The PCR-MGE method is useful not only for rapid identification ( approximately 1.5 h) of staphylococci in positive blood culture bottles, but also for strain delineation of S. aureus. PMID- 15750078 TI - Comparative typing of Campylobacter jejuni by heat-stable serotyping and PCR based restriction fragment length polymorphism analysis. AB - Campylobacter jejuni has become the most common bacterial cause of human gastroenteritis worldwide. Rapid, discriminatory typing methods are required to identify potential clusters of infections. The major disadvantage of the well evaluated and widely used Penner heat-stable serotyping method is the high level of nontypeability. The correlation of the types determined by the Penner heat stable serotyping method and PCR-based restriction fragment length polymorphism (RFLP) analysis of the lipooligosaccharide (LOS) biosynthesis genes of C. jejuni was studied with 149 C. jejuni strains. Of these strains, 79 were patient strains belonging to 25 Penner serotypes, 60 were nontypeable patient strains, and 10 were reference strains. A 9.6-kb DNA fragment of the LOS gene cluster was amplified and digested with the restriction enzymes HhaI and DdeI. Altogether, 39 different RFLP types (including 30 HhaI profiles and 32 DdeI profiles) were identified. Type Hh1Dd1 was the most common type, with 36% of the strains and strains of 12 serotypes being of this type. A high level of discrimination was obtained, and a correlation between the Penner serotypes and the PCR-RFLP types could be seen. Also, variation in the LOS biosynthesis genes within a single Penner serotype was found. Although the PCR-RFLP method may not be sufficient to compensate for Penner serotyping, it can give valuable information about nontypeable strains and further characterize strains of common serotypes. PMID- 15750077 TI - Vi antigen expression in Salmonella enterica serovar Typhi clinical isolates from Pakistan. AB - The accurate identification of Salmonella enterica subsp. enterica serovar Typhi variants that fail to express the capsular polysaccharide, Vi, is an important and much discussed issue for medical microbiology. We have tested a multiplex PCR method which shows the presence or absence of the genetic locus required for Vi expression. Of 2,222 Salmonella serovar Typhi clinical isolates collected from patients' blood over a 4-year period in a region of Pakistan where typhoid is endemic, 12 tested negative for Vi expression by serological agglutination. However, only 1 of these 12 was Vi negative by the multiplex PCR method. This result was confirmed by immunofluorescence, the most sensitive method for Vi characterization in Salmonella serovar Typhi. The multiplex PCR described therefore represents a simple and accurate method for surveillance for Vi negative variants of Salmonella serovar Typhi in Pakistan. Testing of clinical isolates of Salmonella serovar Typhi, before subculture, from other regions where Vi-negative Salmonella serovar Typhi has been described should be carried out so that the impact of vaccination with purified Vi antigen on the levels of Vi negative Salmonella serovar Typhi in bacterial populations can be assessed. PMID- 15750079 TI - Potential limitations of the 16S-23S rRNA intergenic region for molecular detection of Bartonella species. AB - PCR targeting the 16S-23S rRNA gene intergenic transcribed spacer (ITS) region has been proposed as a rapid and reliable method for the detection of Bartonella species DNA in clinical samples. Because of variation in ITS sequences among Bartonella species, a single PCR amplification can be used to detect different species within this genus. Therefore, by targeting the ITS region, multiple PCRs or additional sample-processing steps beyond the primary amplification can be avoided when attempting to achieve molecular diagnostic detection of Bartonella species. Although PCR amplification targeting this region is considered highly sensitive, amplification specificity obviously depends on primer design. We report evidence of nonspecific PCR amplification of Mesorhizobium species with previously published primers that were designed to amplify the Bartonella consensus ITS region. Use of these or other, less species-specific, primers could lead to a false-positive diagnostic test result when evaluating clinical samples. We also report the presence of Mesorhizobium species DNA as a contaminant in molecular-grade water, a series of homologous sequences in the ITS region that are common to Bartonella and Mesorhizobium species, the amplification of Mesorhizobium DNA with unpublished primers designed in our laboratory targeting the ITS region, and the subsequent design of unambiguous ITS primers that avoid nonspecific amplification of Mesorhizobium species. Our results define some potential limitations associated with the molecular detection of Bartonella species in patient samples and indicate that primer specificity is of critical importance if the ITS region is used as a diagnostic target for detection of Bartonella species. PMID- 15750080 TI - Molecular subtyping and characterization of bovine and human Streptococcus agalactiae isolates. AB - Streptococcus agalactiae causes severe invasive disease in humans and mastitis in cattle. Temporally matched bovine milk isolates and clinical human invasive isolates (52 each) collected in New York State over 18 months were characterized by molecular subtyping and phenotypic methods to probe the interspecies transmission potential of this species. EcoRI ribotyping differentiated 17 ribotypes, and DNA sequencing of the housekeeping gene sodA and the putative virulence gene hylB differentiated 7 and 17 allelic types, respectively. Human and bovine isolates were not randomly distributed between ribotypes or hylB and sodA clusters. The combined analysis of all subtyping data allowed the differentiation of 39 clonal groups; 26 groups contained only bovine isolates, and 2 groups contained both human and bovine isolates. The EcoRI ribotype diversity among bovine isolates (Simpson's numerical index of discrimination [mean +/- standard deviation], 0.90 +/- 0.05) being significantly higher than that among human isolates (0.42 +/- 0.15) further supports that these isolates represent distinct populations. Eight human isolates, but no bovine isolates, showed an IS1548 transposon insertion in hylB, which encodes a hyaluronidase. Based on data for 43 representative isolates, human isolates, on average, showed lower hyaluronidase activities than bovine isolates. Isolates with the IS1548 insertion in hylB showed no hyaluronidase activity. Human and bovine isolates did not differ in their abilities to invade HeLa human epithelial cells. Our data show that (i) EcoRI ribotyping, combined with hylB and sodA sequencing, provides a discriminatory subtype analysis of S. agalactiae; (ii) most human invasive and bovine S. agalactiae isolates represent distinct subtypes, suggesting limited interspecies transmission; and (iii) hyaluronidase activity is not required for all human infections. PMID- 15750081 TI - Clinical evaluation of the DiversiLab microbial typing system using repetitive sequence-based PCR for characterization of Staphylococcus aureus strains. AB - The DiversiLab System, which includes microfluidics-based detection, reagent kits, and software for data processing and analysis, is an automated method using repetitive sequence-based PCR (rep-PCR) for microbial strain typing. To assess the reliability of the DiversiLab System for strain characterization of Staphylococcus aureus, we tested clinical isolates sent to ARUP Laboratories for typing and compared results to those of pulsed field electrophoresis (PFGE) aided by the cluster analysis provided by BioNumerics software. spa typing was performed when the results of these two methods for an outbreak were not concordant. The study included 89 S. aureus isolates (65 mecA positive, 24 mecA negative) from 19 outbreaks (2 to 11 isolates/outbreak). The DiversiLab and PFGE BioNumerics results were concordant for 15 of the 19 outbreaks. For the remaining four outbreaks, there was partial concordance between the two methods. spa typing results were the same as or more similar to rep-PCR results for three of those outbreaks and were more similar to PFGE results for one. With regard to performance, the DiversiLab system was considerably less labor intensive than PFGE and provided results in less than 24 h, compared with 2 to 3 days for PFGE. Additionally, the Web-based DiversiLab software provides standardized comparisons among isolates almost instantaneously and generates user-friendly, customized reports. PMID- 15750082 TI - Risk factors associated with colonization by pneumococci with reduced susceptibility to fluoroquinolones in adult outpatients. AB - We developed a case-control study in order to identify risk factors associated with pharyngeal colonization by Streptococcus pneumoniae with reduced susceptibility to fluoroquinolones (ciprofloxacin MIC, > or =4 microg/ml). A total of 400 patients were studied for colonization by quinolone-nonsusceptible S. pneumoniae (QNSP) isolates and risk factors for this colonization. Isolate susceptibility was determined by the agar dilution method. Forty patients were colonized by QNSP (case patients), and 360 patients were not colonized by QNSP (control patients). The MIC range of ciprofloxacin for QNSP isolates was 4 to 8 microg/ml. No isolates were resistant to levofloxacin and moxifloxacin. Risk factors significantly associated with QNSP colonization, according to univariate analysis, were recent hospitalizations (odds ratio [OR], 3.43; 95% confidence interval [CI], 1.6 to 7.2; P < 0.01) and prior exposure to fluoroquinolones (OR, 6.04; 95% CI, 3.0 to 12.0; P < 0.01). Other factors such as chronic obstructive pulmonary disease (OR, 1.94; 95% CI; 0.7 to 5.0), prior exposure to penicillins (OR, 1,68; 95% CI, 0.8 to 3.3) and prior exposure to macrolides (OR 2; 95% CI, 0.6 to 6.2) were more frequent among patients colonized with QNSP, but there was no statistical significance. Multivariate analysis showed that exposure to fluoroquinolones was the only independent factor associated with colonization by QNSP (OR, 4.2; 95% CI, 1.8 to 9.4; P < 0.01). Throat colonization by QNSP is becoming frequent, though most of these isolates (all the isolates in this case) remain susceptible to newer fluoroquinolones. Previous treatment with fluoroquinolones seems to be the main risk factor associated with colonization by QNSP. PMID- 15750084 TI - Analysis of Salmonella enterica serotype Typhi pulsed-field gel electrophoresis patterns associated with international travel. AB - Typhoid fever is a significant cause of morbidity and mortality worldwide, causing an estimated 16 million cases and 600,000 deaths annually. Although overall rates of the disease have dramatically decreased in the United States, the number of travel-related infections has increased in recent decades. Drug resistance among Salmonella enterica serotype Typhi strains has emerged worldwide, making antimicrobial susceptibility testing an important function in public health laboratories. Pulsed-field gel electrophoresis (PFGE) subtyping of food-borne and waterborne pathogens has proven to be a valuable tool for the detection of outbreaks and laboratory-based surveillance. This retrospective study examined the distribution of PFGE patterns of S. enterica serotype Typhi isolates from patients with a history of international travel. Isolates were collected as part of a passive laboratory-based antimicrobial susceptibility surveillance study. Isolates were PFGE subtyped by using the restriction enzyme XbaI to restrict the total genomic DNA. Isolates indistinguishable with XbaI were further characterized using the restriction enzyme BlnI. A total of 139 isolates were typed, representing travel to 31 countries. Restriction fragment patterns consisted of 14 to 18 fragments ranging in size from 580 to 40 kbp. Seventy-nine unique PFGE patterns were generated using XbaI. Isolates from the same geographic region did not necessarily have similar PFGE patterns. Of the 139 isolates, 46 (33%) were resistant to more than one antimicrobial agent (multidrug resistant [MDR]). Twenty-seven (59%) of 46 MDR isolates had indistinguishable PFGE patterns with both XbaI and BlnI. It appears that MDR S. enterica serotype Typhi has emerged as a predominant clone in Southeast Asia and the Indian subcontinent. PMID- 15750083 TI - Molecular characterization of an epidemic clone of panantibiotic-resistant Pseudomonas aeruginosa. AB - We describe the molecular characterization of a multiresistant Pseudomonas aeruginosa clone causing an outbreak in the intensive care unit (ICU) of a tertiary-care university hospital. Analysis included antimicrobial susceptibility profile, O-serotyping, pulsed-field gel electrophoresis, and amplified fragment length polymorphism. Resistance mechanisms were characterized, including production of naturally occurring and acquired beta-lactamases, porin expression, and efflux pump systems. Eighteen patients were colonized or infected with multiresistant P. aeruginosa. Multiresistant P. aeruginosa was panresistant to penicillins, cephalosporins, carbapenems, aminoglycosides, and fluoroquinolones and remained susceptible only to colistin. Sixteen isolates (89%) belonged to serotype O:11, pulsed-field gel electrophoresis type A1, and amplified fragment length polymorphism type A. Resistance characterization of this epidemic clone showed an overexpression of the chromosomal cephalosporinase AmpC combined with decreased expression of porin OprD and the absence of metallo-beta-lactamase or extended-spectrum beta-lactamase. An upregulation of the MexXY efflux system due to an agrZ mutation in the mexZ repressor was detected. This epidemic clone was restricted to the ICU and was not found elsewhere in hospital. Contamination of the ICU environment and the hands of an ICU nurse with this clone suggests possible hand-borne transmission. Implementation of contact precautions effectively controlled transmission of the epidemic clone. This study illustrates the ability of multiresistant P. aeruginosa to cause an outbreak with significant morbidity and mortality and underscores the need to identify clonal outbreaks, which require targeted infection control measures. PMID- 15750085 TI - CHROMagar Candida as the sole primary medium for isolation of yeasts and as a source medium for the rapid-assimilation-of-trehalose test. AB - The chromogenic medium BBL CHROMagar Candida (CAC) was evaluated as a sole primary medium for the isolation of yeasts from clinical specimens in which yeasts are the primary concern. Additionally, the reliability of the rapid assimilation-of-trehalose (RAT) test in yielding correct results with isolates taken from CAC was assessed. A total of 270 throat, urine, and genital (TUG) specimens were streaked onto CAC, Sabouraud dextrose agar (SDA), inhibitory mold agar (IMA), and Mycosel (MYC). A total of 69 blood culture broths that were smear positive for yeast were streaked onto CAC and SDA. A 1-h RAT test (NCCLS M35-A) was performed simultaneously on isolates from CAC and SDA. A total of 112 TUG specimens yielded yeast colonies (CAC, 111 colonies; IMA, 105; SDA, 103; MYC, 91). The 69 blood culture yeasts grew on both CAC and SDA. Mixed cultures of yeasts were detected on 11 CAC plates but were unrecognized on other media. Colonies suspected of being C. glabrata on 32 CAC plates were all RAT test positive and confirmed to be C. glabrata; of 59 colonies with various characteristics of color and morphology on CAC, none were RAT positive, and all were conventionally identified as yeasts other than C. glabrata (sensitivity and specificity, 100%). The same isolates from SDA tested for RAT produced six false negatives and no false positives (sensitivity, 81%; specificity, 100%). The results show that CAC can be used as the sole primary medium for recovery of yeasts from clinical specimens. Additionally, isolates grown on CAC yield excellent results with the RAT test utilized in this study. PMID- 15750086 TI - Seroepidemiology of human metapneumovirus (hMPV) on the basis of a novel enzyme linked immunosorbent assay utilizing hMPV fusion protein expressed in recombinant vesicular stomatitis virus. AB - Human metapneumovirus (hMPV) is a newly identified human respiratory virus now recognized as a major respiratory pathogen of infants and children. To define the seroepidemiology of hMPV, we developed a novel enzyme-linked immunosorbent assay (ELISA) based on expression of the fusion protein of hMPV (hMPV F) in recombinant vesicular stomatitis virus (VSV). Western blot analysis using an hMPV F-specific antiserum confirmed the expression of hMPV in recombinant VSV. The ELISA is specific for hMPV F; antibody specific for the most closely related human paramyxovirus, respiratory syncytial virus, does not bind to hMPV F. Overall, 216 serum specimens were tested. The percentages of seropositive individuals were 89.1% in children < or =5 months old, 55.0% in children 6 to 11 months old, 36.0% in children 12 to 23 months old, 45.0% in children 24 to 47 months old, 77.3% in children 48 to 59 months old, 91.3% in children 5 to 10 years old, and 95.5% for individuals 11 to 20 years old. This is the first seroepidemiological survey of hMPV in the United States and the first analysis to determine the prevalence of antibody to a specific hMPV protein. The data suggest that exposure to hMPV is common in childhood and that hMPV F is an antigenic determinant of hMPV. PMID- 15750087 TI - Systematic molecular characterization of multidrug-resistant Mycobacterium tuberculosis complex isolates from Spain. AB - We used spoligotyping and restriction fragment length polymorphism (RFLP) of the IS6110-insertion sequence to study the molecular epidemiology of multidrug resistant (MDR) tuberculosis in Spain. We analyzed 180 Mycobacterium tuberculosis complex isolates collected between January 1998 and December 2000. Consecutive isolates from the same patients (n = 23) always had identical genotypes, meaning that no cases of reinfection occurred. A total of 105 isolates (58.3%) had unique RFLP patterns, whereas 75 isolates (41.7%) were in 20 different RFLP clusters. Characterization of the katG and rpoB genes showed that 14 strains included in the RFLP clusters did not actually cluster. Only 33.8% of the strains isolated were suggestive of MDR transmission, a frequency lower than that for susceptible strains in Spain (46.6%). We found that the Beijing/W genotype, which is prevalent worldwide, was significantly associated with immigrants. The 22 isolates in the largest cluster corresponded to the Mycobacterium bovis strain responsible for two nosocomial MDR outbreaks in Spain. PMID- 15750088 TI - Epidemiologic import of tuberculosis cases whose isolates have similar but not identical IS6110 restriction fragment length polymorphism patterns. AB - Isolates of Mycobacterium tuberculosis from patients with epidemiologic links frequently demonstrate identical IS6110 restriction fragment length polymorphism (RFLP) patterns (i.e., RFLP clustering) because they are infected with the same strain. Uncertainty arises with isolates that differ from one another by a few IS6110 hybridizing bands. During the period from 1 January 1996 to 31 December 1999, isolates from 585 tuberculosis (TB) cases were analyzed by RFLP, representing 98.2% of the 596 culture-positive TB cases reported in Arkansas during the study period. Of the 585 cases for which RFLP was available, 419 (71.6%) had an RFLP pattern with more than five copies of IS6110. Of the total 74 clusters, 48 comprised isolates with more than five copies of IS6110 and included 164 cases. Sixty-nine isolates with more than five copies of IS6110 comprising 16 clusters and 60 unique isolates were found to be similar to at least 1 other isolate (differing from it by one or two hybridizing bands). Among the 129 cases whose isolates were similar to other clustered or unique isolates, 16 cases were discovered with epidemiologic links: 14 (15.2%) were among the 92 cases with IS6110 RFLP patterns similar to those in clusters, and 2 (5.2%) were among the 37 unique cases that were similar to another unique case. The isolates from the epidemiologically linked patients shared common spoligotypes; all except one case shared common polymorphic GC-rich sequence (PGRS) patterns. Of the 129 patients whose isolates differed from another by one or two hybridizing IS6110 bands, 101 (78.3%) shared common spoligotypes and 87 (67.4%) shared common PGRS RFLP patterns. PMID- 15750089 TI - Identification of Streptococcus canis isolated from milk of dairy cows with subclinical mastitis. AB - Streptococcus canis was isolated from 31 milk samples from 11 cows in a dairy herd (with 49 lactating cows) affected by subclinical mastitis in north Rhine Westphalia, Germany. Thirty-one isolates from the infected udder quarters were further characterized for their phenotypic and molecular properties. Most isolates (83.9%) produced alpha-galactosidase, and all were negative for beta-d glucuronidase. Amplification of the 16S rRNA gene by the PCR method and digestion with the restriction enzymes RsaI, MspI, and AvaII yielded species-specific patterns. Additional identification by species-specific amplification of the 16S rRNA gene, the 16S-23S rRNA gene intergenic spacer region, the CAMP factor encoding gene cfg, and the internal fragments of the sodA gene was consistent with S. canis. Macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis showed that the S. canis isolates originated from a single clone or were very closely related. PMID- 15750090 TI - Detection of human picornaviruses by multiplex reverse transcription-PCR and liquid hybridization. AB - A qualitative multiplex reverse transcription (RT)-PCR and liquid hybridization assay for the detection of human enteroviruses, rhinoviruses, parechoviruses, and Aichi virus was developed. Furthermore, a separate assay for the recognition of hepatitis A virus was established to complement the test pattern so that all human picornaviruses were covered. The amplicons, which represented the 5' untranslated regions of the viral RNA genomes, were identified in liquid hybridization reactions with genus-specific digoxigenin-labeled oligonucleotide probes. The sensitivity of the multiplex RT-PCR and liquid hybridization assay was 10 to 100 picornavirus genome equivalents for representatives of each picornavirus genus. The hepatitis A virus assay exhibited a sensitivity of 10 genome copies. Both the uniplex and the multiplex tests were highly specific for the target viruses. Twenty-three clinical samples, including cerebrospinal fluid, serum, and nasopharyngeal swab specimens, were used for clinical evaluation of the multiplex RT-PCR assay. The results obtained were consistent with the results of routine virus diagnostic assays. Furthermore, the assay was used to screen 68 stool specimens for the presence of parechoviruses and Aichi virus. One sample was found to contain parechovirus RNA, whereas no Aichi virus was detected. The assay described here can be applied for the efficient identification of human enteroviruses and rhinoviruses in clinical specimens and simultaneously enables the collection of information on the epidemiology and clinical outcomes of infections caused by the currently poorly known human parechoviruses and Aichi virus. PMID- 15750091 TI - Characterization of virulence plasmids and serotyping of rhodococcus equi isolates from submaxillary lymph nodes of pigs in Hungary. AB - The plasmid types and serotypes of 164 Rhodococcus equi strains obtained from submaxillary lymph nodes of swine from different piggeries in 28 villages and towns located throughout the country were examined. The strains were tested by PCR for the presence of 15- to 17-kDa virulence-associated protein antigen (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphism characteristics. None of the 164 isolates contained the vapA gene, and 44 (26.8%) isolates were positive for the vapB gene, showing a product of the expected 827-bp size in the PCR amplification. The 44 isolates of intermediate virulence contained virulence plasmids that were identified as types 1 (3 isolates), 4 (1 isolate), 5 (36 isolates), 6 (1 isolate), and 7 (2 isolates) and as a new variant (1 isolate). On the basis of restriction digestion patterns of plasmid DNAs, we tentatively designated the variant as type 17. Use of the serotyping method of Prescott showed that 110 (67.1%) out of the 164 isolates were typeable and that serotype 2 predominated (83 isolates [50.6%]), followed by serotype 1 (26 strains [15.9%]). Only one isolate belonged to serotype 3. A total of 54 (32.9%) isolates were untypeable in Prescott's system. The prevalence of R. equi strains of intermediate virulence among the isolates that came from the submaxillary lymph nodes of swine in Hungary was lower than that seen with isolates obtained elsewhere. PMID- 15750092 TI - Analysis of the influence of Tween concentration, inoculum size, assay medium, and reading time on susceptibility testing of Aspergillus spp. AB - The influence of several test variables on susceptibility testing of Aspergillus spp. was assessed. A collection of 28 clinical isolates was tested against amphotericin B, itraconazole, voriconazole, and terbinafine. Inoculum size (10(4) CFU/ml versus 10(5) CFU/ml) and glucose supplementation (0.2% versus 2%) did not have significant effects on antifungal susceptibility testing results and higher inoculum size and glucose concentration did not falsely elevate MICs. In addition, antifungal susceptibility testing procedure with an inoculum size of 10(5) CFU/ml distinctly differentiated amphotericin B or itraconazole-resistant Aspergillus strains in vivo from the susceptible ones. Time of incubation significantly affected the final values of MICs, showing major increases (two to six twofold dilutions, P < 0.01 by analysis of variance) between MIC readings at 24 and 48 h, but no differences were observed between antifungal susceptibility testing results obtained at 48 h and at 72 h. Significantly higher MICs were uniformly associated with higher concentrations of Tween (P < 0.01), used as a dispersing agent in the preparation of inoculum suspensions. The geometric mean MICs showed increases of between 1.5- and 10-fold when the Tween concentration varied from 0.1% (the geometric means for amphotericin B, itraconazole, voriconazole, and terbinafine were 1.29, 0.69, 1.06, and 0.64 mug/ml, respectively) to 5% (the geometric means for amphotericin B, itraconazole, voriconazole, and terbinafine were 1.97, 5.79, 1.60, and 4.66 mug/ml, respectively). The inhibitory effect of Tween was clearly increased with inoculum sizes of 10(5) CFU/ml and was particularly dramatic for itraconazole, terbinafine, and Aspergillus terreus. The inoculum effect was not observed when the Tween concentration was below 0.5% (P > 0.01). PMID- 15750093 TI - Multiplex real-time PCR assay using Scorpion probes and DNA capture for genotype specific detection of Giardia lamblia on fecal samples. AB - Two major genotypic assemblages of Giardia lamblia infect humans; the epidemiologic significance of this phenomenon is poorly understood. We developed a single-vessel multiplex real-time PCR (qPCR) assay that genotypes Giardia infections into assemblages A and/or B directly from fecal samples. The assay utilized Scorpion probes that combined genotype-specific primers and probes for the 18S rRNA gene into the same molecule. The protocol was capable of detecting as few as 20 trophozoites per PCR on fecal DNA isolated using a commercial method or 1.25 trophozoites per PCR on fecal DNA isolated using a G. lamblia-specific oligonucleotide capture technique. The assay was specific for fecal specimens, with no amplification of the discordant genotype with the opposite Scorpion probe. When 97 clinical specimens from Bangladesh were used, the multiplex PCR assay detected 95% (21 of 22) of Giardia microscopy-positive specimens and 18% (13 of 74) of microscopy-negative specimens. Microscopy-negative and qPCR positive specimens had higher average cycle threshold values than microscopy positive and qPCR-positive specimens, suggesting that they represented true low burden infections. Most (32 of 35) infections were assemblage B infections. This single-reaction multiplex qPCR assay distinguishes assemblage A Giardia infections from assemblage B infections directly on fecal samples and may aid epidemiologic investigation. PMID- 15750094 TI - Distribution of Mycobacterium avium complex isolates in tissue samples of pigs fed peat naturally contaminated with mycobacteria as a supplement. AB - In early 1999, there was an increased incidence of tuberculous lesions in the lymph nodes of slaughtered pigs in the Czech Republic. In part 1 of this study, tuberculous lesions were detected in 140 (62%) tissue samples collected from pigs coming from 15 farms in 15 districts at routine veterinary meat inspections in abattoirs. Mycobacteria were isolated from 37 (16%) tissue samples: 34 Mycobacterium avium subsp. hominissuis isolates and three environmentally derived mycobacteria. In search of infection sources, M. avium subsp. hominissuis was isolated from 38 (79%) samples of peat used as a feed supplement. In part 2 of our study, the head, mesenteric, and inguinal lymph nodes of 117 randomly selected slaughtered pigs from one farm with young piglets fed peat as a supplement were investigated for mycobacterial infection. From 65 (56%) pigs, a total of 76 mycobacterial isolates were identified (56 M. avium subsp. hominissuis isolates, 5 M. avium subsp. avium isolates, 3 M. intracellulare isolates, and 12 environmentally derived mycobacterial isolates). IS1245 restriction fragment length polymorphism (RFLP) types with >20 bands of 45 distinct RFLP types were found in 49 M. avium subsp. hominissuis isolates from pigs (n = 31) and peat (n = 18). Identical RFLP types were found in only four pig isolates. Five randomly selected isolates from pigs and peat were subcultured to six independent clones or colonies. Among the IS1245 RFLP types of 30 clones, identical RFLP types obtained from pigs and peat were identified, which confirmed the hypothesis that peat contaminated with mycobacteria represents a significant source of mycobacterial infection for pigs. PMID- 15750095 TI - Leishmania promastigote membrane antigen-based enzyme-linked immunosorbent assay and immunoblotting for differential diagnosis of Indian post-kala-azar dermal leishmaniasis. AB - Diagnosis of post-kala-azar dermal leishmaniasis (PKDL), caused by Leishmania donovani, is difficult, as the dermal lesions are of several types and resemble those caused by other skin diseases, especially leprosy. Since the disease generally appears very late after the clinical cure of kala-azar in India, it is also difficult to correlate PKDL with a previous exposure to L. donovani. Very few attempts have been made so far to diagnose PKDL serologically, and the diagnostic methods vary in their sensitivities and specificities. Diagnosis of PKDL through sophisticated PCR methods, although highly sensitive, has limited practical use. We have developed a serodiagnostic method using an enzyme-linked immunosorbent assay to detect specific immunoglobulin (Ig) isotypes and IgG subclass antibodies in the sera of Indian PKDL patients. Our assay, which uses L. donovani promastigote membrane antigens, was 100% sensitive for the detection of IgG and 96.7% specific for the detection of IgG and IgG1. Optical density values for individual patients, however, demonstrated wide variations. Western blot analysis based on IgG reactivity could differentiate patients with PKDL from control subjects, which included patients with leprosy, patients from areas where kala-azar is endemic, and healthy subjects, by the detection of polypeptides of 67, 72, and 120 kDa. The recognition patterns of the majority of serum samples from patients with PKDL were also distinct from those of the serum samples from patients with visceral leishmaniasis (VL), at least for a 31-kDa polypeptide. To further differentiate patients with PKDL from those with active and cured VL, we analyzed the specific titers of the Ig isotypes and IgG subclasses. High levels of IgG, IgG1, IgG2, and IgG3 antibodies significantly differentiated patients with PKDL from patients cured of VL. The absence of antileishmanial IgE and IgG4 in patients with PKDL differentiated these patients from those with active VL. These results imply intrinsic differences in the antibodies generated in the sera from patients with PKDL and VL. PMID- 15750096 TI - Cytotoxin and pyrogenic toxin superantigen gene profiles of Staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles. AB - A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands-[lukED, seg, sei, sem, sen, seo +/- seu] with Sabeta-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively-while 11.9% of them carried both islands. Only one contained seb (together with upsilonSabeta-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Sabeta-1, SaPIbov, or both. Five SmaI profiles (S > or = 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (upsilonSabeta-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis. PMID- 15750097 TI - Trends in drug resistance, serotypes, and molecular types of Streptococcus pneumoniae colonizing preschool-age children attending day care centers in Lisbon, Portugal: a summary of 4 years of annual surveillance. AB - Of the nasopharyngeal cultures recovered from 942 day care center (DCC) attendees in Lisbon, Portugal, 591 (62%) yielded Streptococcus pneumoniae during a surveillance performed in February and March of 1999. Forty percent of the isolates were resistant to one or more antimicrobial agents. In particular, 2% were penicillin resistant and 20% had intermediate penicillin resistance. Multidrug resistance to macrolides, lincosamides, and tetracycline was the most frequent antibiotype (17% of all isolates). Serotyping and molecular typing by pulsed-field gel electrophoresis were performed for 202 out of 237 drug-resistant pneumococci (DRPn). The most frequent serotypes were 6B (26%), 14 (22%), 19F (16%), 23F (10%), and nontypeable (12%). The majority (67%) of the DRPn strains were representatives of nine international clones included in the Pneumococcal Molecular Epidemiology Network; eight of them had been detected in previous studies. Fourteen novel clones were identified, corresponding to 26% of the DRPn strains. The remaining 7% of the strains were local clones detected in our previous studies. Comparison with studies conducted since 1996 in Portuguese DCCs identified several trends: (i) the rate of DRPn frequency has fluctuated between 40 and 50%; (ii) the serotypes most frequently recovered have remained the same; (iii) nontypeable strains appear to be increasing in frequency; and (iv) a clone of serotype 33F emerged in 1999. Together, our observations highlight that the nasopharynxes of children in DCCs are a melting pot of successful DRPn clones that are important to study and monitor if we aim to gain a better understanding on the epidemiology of this pathogen. PMID- 15750098 TI - Molecular epidemiology of macrolide-resistant Streptococcus pneumoniae isolates in Europe. AB - In many European countries, the level of pneumococcal resistance to macrolides has now passed the level of resistance to penicillin G. A total of 82 erythromycin A-resistant isolates of Streptococcus pneumoniae were collected by 11 laboratories in seven European countries. All of the isolates were tested for antimicrobial susceptibility, analyzed for clonal relatedness by multilocus sequence typing, and characterized for macrolide resistance genotypes. The prevalence of the macrolide resistance genotypes varied substantially between countries. In France (87.5% of all strains), Spain (77.3%), Switzerland (80%), and Poland (100%), strains were predominantly erm(B) positive, whereas higher levels of mef(A)-positive strains were reported from Greece (100%) and Germany (33.3%). Macrolide resistance was caused by the oligoclonal spread of some multilocus sequence types, but significant differences in clonal distribution were noted between France and Spain, countries from which high levels of macrolide resistance have been reported. Overall, sequence type 81 (Spain23F-1 clone) was by far the most widespread. The mainly erm(B)-positive serotype 14 clone (sequence type 143), first reported in Poland in the mid-1990s, is now widespread in France. PMID- 15750099 TI - Comparison of Easy-Flow Copan Liquid Stuart's and Starplex Swab transport systems for recovery of fastidious aerobic bacteria. AB - Because samples are frequently submitted on swabs from distant sites, viability of the organism must be maintained. We compared two transport systems, a new Copan Liquid Stuart's swab with an Easy-Flow swab applicator and the Starplex Liquid Stuart's swab. The purpose of the study was to assess the release and/or recovery of organisms from the Copan system compared to that from Starplex. Triplicate swabs were seeded with 3 dilutions of Neisseria gonorrhoeae, Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae. Although the amount of the initial inoculum was the same for both transport systems, recovery by the roll-plate method at time zero was consistently increased with the Copan system (31 to 87% higher). This is the most important finding in this study. With N. gonorrhoeae, subsequent recoveries were similar for Copan and Starplex but poor for both systems. With N. meningitidis and Haemophilus, higher levels of recovery were clearly obtained with Copan (P < 0.05 to P < 0.001). With Streptococcus, subsequent recoveries for Copan and Starplex were mixed. In conclusion, Copan generally demonstrated better recovery of organisms compared to Starplex even (and especially) at time zero. PMID- 15750100 TI - Evaluation of a point-of-care test, BVBlue, and clinical and laboratory criteria for diagnosis of bacterial vaginosis. AB - Bacterial vaginosis (BV) remains the most common cause of abnormal vaginal discharge in women of reproductive age and is associated with increased susceptibility to human immunodeficiency virus and sexually transmitted infections and preterm delivery. Present diagnostic methods require access to microscopy and laboratory expertise; however, the majority of women, particularly those in populations with a high prevalence of BV, do not have access to clinical services with on-site microscopy capabilities. We evaluated a point-of-care test for the diagnosis of BV, the BVBlue test, with 288 women attending a sexual health service with symptoms of abnormal vaginal discharge and/or odor. The BVBlue test performed well compared with conventional diagnostic methods for the assessment of women with symptoms suggestive of BV at the bedside and significantly better than other simple tests, such as vaginal pH determination and the amine test, that do not require microscopy. The BVBlue test was sensitive (88%; 95% confidence interval [CI], 81 to 93%) and specific (95%; 95% CI, 91 to 98%) compared to the method of Nugent et al. (R. P. Nugent, M. A. Krohn, and S. L. Hillier, J. Clin. Microbiol. 29:297-301, 1991) and performed well compared with the method of Amsel et al. (R. Amsel, P. A. Totten, C. A. Spiegel, K. C. Chen, D. Eschenbach, and K. K. Holmes, Am. J. Med. 74:14-22, 1983), with a sensitivity of 88% (95% CI, 81 to 93%) and a specificity of 91% (95% CI, 85 to 94%). The BVBlue test is a simple, rapid, and objective test for the diagnosis of BV and has the potential to facilitate prompt diagnosis and appropriate treatment of BV in the absence of microscopy. The majority of women at the greatest risk for the sequelae of BV are not in settings where the conventional diagnostic methods are either practical or possible, and they would greatly benefit from access to rapid and reliable point-of-care tests to improve the diagnosis and management of BV. PMID- 15750101 TI - Sequence analysis of the msp4 gene of Anaplasma phagocytophilum strains. AB - The causative agent of human granulocytic ehrlichiosis was recently reclassified as Anaplasma phagocytophilum, unifying previously described bacteria that cause disease in humans, horses, dogs, and ruminants. For the characterization of genetic heterogeneity in this species, the homologue of Anaplasma marginale major surface protein 4 gene (msp4) was identified, and the coding region was PCR amplified and sequenced from a variety of sources, including 50 samples from the United States, Germany, Poland, Norway, Italy, and Switzerland and 4 samples of A. phagocytophilum-like organisms obtained from white-tailed deer in the United States. Sequence variation between strains of A. phagocytophilum (90 to 100% identity at the nucleotide level and 92 to 100% similarity at the protein level) was higher than in A. marginale. Phylogenetic analyses of msp4 sequences did not provide phylogeographic information but did differentiate strains of A. phagocytophilum obtained from ruminants from those obtained from humans, dogs, and horses. The sequence analysis of the recently discovered A. phagocytophilum msp2 gene corroborated these results. The results reported here suggest that although A. phagocytophilum-like organisms from white-tailed deer may be closely related to A. phagocytophilum, they could be more diverse. These results suggest that A. phagocytophilum strains from ruminants could share some common characteristics, including reservoirs and pathogenicity, which may be different from strains that infect humans. PMID- 15750102 TI - Human cytomegalovirus reactivation during lactation and mother-to-child transmission in preterm infants. AB - In a clinical trial, the incidence of cytomegalovirus reactivation in breastfeeding mothers and transmission to their preterm infants were studied. Breast milk from 73 mothers as well as urine and tracheal and pharyngeal aspirates from their 89 infants were screened weekly for human cytomegalovirus (HCMV) DNA during the first 2 months after delivery. Of the 73 mothers, 48 (66%) were positive for HCMV DNA in the lactating breast. HCMV reactivation could be confirmed for 19 of 20 (95%) immunoglobulin G-positive mothers. Of the eight immunoglobulin G-negative mothers one was positive for HCMV DNA in breast milk. In only 2 out of 13 seropositive mothers with HCMV DNA in breast milk could viral DNA be detected in the peripheral blood. HCMV mother-to-child transmission was concluded for 20 of the 48 (42%) mothers positive for DNA or 7 of 19 (37%) seropositive for HCMV and positive for HCMV DNA in breast milk and one of one mother seronegative for HCMV but positive for HCMV DNA in breast milk. One mother transmitted the virus to her twins. In addition, one infant acquired postnatal HCMV infection despite the mother's being negative for HCMV DNA in breast milk; altogether, we found 22 infants with HCMV infection. In 13 of these 22 infants, virus infection occurred definitively postnatally; two of them developed severe symptomatic HCMV infection. HCMV-infected infants demonstrated higher incidences of amniotic infection, respiratory distress syndrome, bronchopulmonary dysplasia, and retinopathia praenatalis than noninfected infants, however, the differences were not statistically significant. In summary, our study confirmed a very high incidence of HCMV reactivation in mothers during lactation and a significant risk of transmission to preterm infants with the possibility of severe disease in these babies. PMID- 15750103 TI - Effect of clarithromycin treatment on Chlamydia pneumoniae in vascular tissue of patients with coronary artery disease: a randomized, double-blind, placebo controlled trial. AB - Several small clinical trials have indicated that antibiotic treatment of Chlamydia pneumoniae infection is associated with a better outcome in patients with coronary artery disease (CAD). It has not been demonstrated whether antibiotic treatment eradicates C. pneumoniae from vascular tissue. The aim of the present study was to assess the effect of clarithromycin on the presence of C. pneumoniae in the vascular tissue of patients with CAD. Patients who had CAD and who were waiting for coronary artery bypass graft surgery were enrolled in a randomized, double-blind, placebo-controlled trial. Patients were treated with clarithromycin at 500 mg or placebo once daily from the day of inclusion in the study until surgery. Several vascular tissue specimens were obtained during surgery. The presence of C. pneumoniae in vascular tissue specimens was examined by immunohistochemical staining (IHC) and two PCR assays. Chlamydia immunoglobulin G (IgG) titers were determined by an enzyme-linked immunosorbent assay at the time of inclusion in the study and 8 weeks after surgery. A total of 76 patients were included, and 180 vascular tissue specimens were obtained (80 specimens from the group treated with clarithromycin and 100 specimens from the group treated with placebo). Thirty-five patients received clarithromycin (mean duration, 27 days; standard deviation [SD], 12.2 days), and 41 patients received placebo (mean duration, 27 days; SD, 13.9 days). IHC detected the C. pneumoniae major outer membrane protein antigen in 73.8% of the specimens from the group treated with clarithromycin and 77.0% of the specimens from the group treated with placebo (P was not significant). Chlamydia lipopolysaccharide antigen was found in only one specimen from the group that received placebo. C. pneumoniae DNA was not detected in any specimen. Baseline Chlamydia-specific IgG titers were equally distributed in both groups and were not significantly different after treatment. There was no indication of an active C. pneumoniae infection in vascular tissue. Chlamydia-specific IgG titers remained unchanged throughout the study in both the antibiotic- and the placebo-treated patients. PMID- 15750104 TI - Systemic disease in Vaal rhebok (Pelea capreolus) caused by mycoplasmas in the mycoides cluster. AB - In the winter of 2002, an outbreak of mycoplasma infection in Vaal rhebok (Pelea capreolus) originating from South Africa occurred 15 weeks after their arrival in San Diego, Calif. Three rhebok developed inappetence, weight loss, lethargy, signs related to pulmonary or arthral dysfunction, and sepsis. All three rhebok died or were euthanized. Primary postmortem findings were erosive tracheitis, pleuropneumonia, regional cellulitis, and necrotizing lymphadenitis. Mycoplasmas were detected in numerous tissues by electron microscopy, immunohistochemistry, and PCR. The three deceased rhebok were coinfected with ovine herpesvirus-2, and two animals additionally had a novel gammaherpesvirus. However, no lesions indicative of herpesvirus were seen microscopically in any animal. The rheboks' mycoplasmas were characterized at the level of the 16S rRNA gene, the 16S-23S intergenic spacer region, and the fructose biphosphate aldolase gene. Denaturing gradient gel electrophoresis was carried out to address the possibility of infection with multiple strains. Two of the deceased rhebok were infected with a single strain of Mycoplasma capricolum subsp. capricolum, and the third animal had a single, unique strain most closely related to Mycoplasma mycoides subsp. mycoides large-colony. A PCR survey of DNA samples from 46 other ruminant species demonstrated the presence of several species of mycoplasmas in the mycoides cluster, including a strain of M. capricolum subsp. capricolum identical to that found in two of the rhebok. These findings demonstrate the pervasiveness of mycoplasmas in the mycoides cluster in small ruminants and the potential for interspecies transmission and disease when different animal taxa come in contact. PMID- 15750105 TI - External quality assessment for detection of Chlamydia trachomatis. AB - The use of molecular methods for detection of Chlamydia trachomatis is increasing in clinical laboratories. External quality assessment enables unbiased monitoring of the performance of laboratories in the detection of specific pathogens. This study details the results of molecular and enzyme immunosorbent assay (EIA) testing for C. trachomatis detection in simulated endocervical swab specimens recently distributed internationally by United Kingdom National External Quality Assessment Scheme for Microbiology (UK NEQAS for Microbiology) external quality assessment panels. The frequency of accurate detection of C. trachomatis in the panels ranged from 32 to 100%. Participants using molecular methods were significantly more likely to detect C. trachomatis in specimens than those using an EIA. Two strains were distributed with the panels: an L2 laboratory-adapted strain and an uncharacterized primary isolate. Further analysis indicated a difference in detection of C. trachomatis between specific methods only with the L2 strain at lower concentrations. In addition, eight negative specimens were distributed, and false positives were found to be rare by all methods included in the study. PMID- 15750106 TI - Epidemiology of human sporotrichosis investigated by amplified fragment length polymorphism. AB - Amplified fragment length polymorphism (AFLP) was used to analyze the genetic diversity of Peruvian strains of Sporothrix schenckii and to compare them to a panel of non-Peruvian strains. AFLP analysis suggests that the Peruvian strains can be divided into two homogeneous clusters with no reference to geographical origin or the clinical form of sporotrichosis. The strains from abroad present heterogeneous profiles, with the Bolivian strain and the Colombian strains related to one of the Peruvian population. Sequencing of internal transcribed spacer 2, used to examine the relationships over a longer distance, confirmed the division of Peruvian strains into two populations that can be identified on the basis of a single but specific sequence divergence. This paper introduces automated AFLP analysis as a valuable tool for further investigation of the epidemiology and ecology of S. schenckii. PMID- 15750108 TI - Loss of the mecA gene during storage of methicillin-resistant Staphylococcus aureus strains. AB - The mecA gene was lost in 36 (14.4%) of 250 methicillin-resistant Staphylococcus aureus isolates after 2 years of storage at -80 degrees C with the Microbank system (Pro-lab Diagnostics, Austin, Tex.). Further analysis of 35 of these isolates confirmed loss of the mecA gene in 32 isolates. This finding has important implications for the management of strain collections. PMID- 15750107 TI - Molecular epidemiology of Shigella flexneri in a long-stay psychiatric nursing center during 2001 to 2003. AB - With six separate wards accommodating more than 1,600 patients, V Nursing Center (VNC) is a long-stay psychiatric nursing center in eastern Taiwan. During 2001 to 2003, 39 shigellosis cases occurred in VNC. Different from the notion that most cases of shigellosis are caused by Shigella sonnei, all except one of these cases were caused by S. flexneri, with the remaining one caused by an S. sonnei isolate. O-antigen serotyping showed that the 38 S. flexneri strains were of either type 1a (n = 20) or 4a (n = 18), two less prevalent serotypes in Taiwan. NotI-based pulsed-field gel electrophoresis analyses performed with 8 type 1a non VNC strains and 9 type 4a non-VNC strains isolated from 1996 to 2003 for comparison divided the 28 type 1a strains and the 27 type 4a strains into 7 and 10 subtypes, designated subtypes P1A to P1G and subtypes P4A to P4J, respectively. Subtypes P1A and P4A, which appeared in three consecutive years in VNC as well as outside of VNC, are the most prevalent subtypes. Analyses of the relatedness of the VNC strains on the basis of the banding patterns grouped the type 1a and 4a strains into four and five clusters, respectively. All except one of the type 1a strains had 95% similarity, indicating that they had a common parent, whereas the type 4a strains had similarities that ranged from 77 to 93%, suggesting that they were of diverse origins. In two of the outbreaks, less related subtypes of the type 4a strains were found in the same VNC wards in consecutive years, suggesting the possible existence of different subtypes in VNC all the time. Antibiotic susceptibility testing showed that all except one of the S. flexneri strains were sensitive to at least seven antibiotics; the remaining isolate was sensitive to three antibiotics. The data from the latter tests should be helpful for selection of proper treatments for S. flexneri infections in Taiwan. PMID- 15750109 TI - Predominance of afr2 and ral fimbrial genes related to those encoding the K88 and CS31A fimbrial adhesins in enteropathogenic Escherichia coli isolates from rabbits with postweaning diarrhea in Central Europe. AB - PCR tests designed in these studies identified three rabbit adhesive factor genes among 43 enteropathogenic E. coli (EPEC) strains: afr1 (2 strains), the F4(K88)/CS31A-related afr2 (10 strains), and ral (15 strains). Several EPEC strains (i.e., O153:H7 and O157:H2) lacked these genes but did adhere to HeLa cells and produced attaching and effacing lesions in rabbits. PMID- 15750110 TI - Confirming positive results of nucleic acid amplification tests (NAATs) for Chlamydia trachomatis: all NAATs are not created equal. AB - The Centers for Disease Control and Prevention recommended confirming positive screening tests for Chlamydia trachomatis when positive predictive values are <90%. It is accepted that less sensitive tests (i.e., culture and immunoassays) should not be used to confirm the results of more sensitive nucleic acid amplification tests (NAATs). We show that the same principle applies when NAATs are used for confirmation. PMID- 15750111 TI - Serotype G9 rotavirus infections in adults in Sweden. AB - Rotavirus is a major cause of acute gastroenteritis. By examining 1,517 stool samples collected in 2001 and 2002 from Swedish adults with acute diarrhea, rotavirus was found in 3.2%, with the emerging G9P[8] serotype being the one most commonly identified (42.9%). This is the first documentation of G9 infections in adults in Europe. PMID- 15750112 TI - Predominance of porcine rotavirus G9 in Japanese piglets with diarrhea: close relationship of their VP7 genes with those of recent human G9 strains. AB - Type G9 of group A rotavirus (GAR) was shown to be predominant in a survey of VP7 (G) and VP4 (P) genotypes among porcine GARs associated with outbreaks of diarrhea in young pigs in Japan between 2000 and 2002. Comparison of the G9 VP7 gene sequences showed that the porcine G9 strains were more closely related to human G9 strains reemerging globally since the mid-1990s than to those from the mid-1980s. The VP7 gene sequences of porcine G9 strains from different farms were divergent (6.1 to 7.2% difference in nucleotides), suggesting that these G9 VP7 genes were not the result of recent introduction into the porcine population. Regarding the P genotype specificities of porcine G9 strains, while the majority of strains were close to unusual porcine P types (P[13] and P[23]), two strains were of the P[6] type, which has closer sequence identity with the human AU19 strain than with the porcine Gottfried strain. These unexpected results suggest that G9 GARs in the porcine population have spread more widely than previously thought and that the VP7 genes of porcine G9 strains and those of some human G9 strains detected recently may have a common progenitor. PMID- 15750113 TI - BK and JC polyomaviruses are not associated with idiopathic pulmonary fibrosis. AB - We sought to determine if the BK and JC polyomaviruses were associated with idiopathic pulmonary fibrosis (IPF). We did not detect the BK or JC polyomaviruses in lung tissue extracts from 33 patients with IPF by using real time PCR, which suggests that an etiologic association is unlikely. PMID- 15750114 TI - Use of microsatellite markers and gene dosage to quantify gene copy numbers in Candida albicans. AB - With microsatellite marker typing, the number of alleles must be known for calculation of allelic frequencies in the diploid Candida albicans for a given locus. We describe a gene dosage with a double real-time PCR. Such a dosage should also be useful in exploring the loss of heterozygosity in C. albicans. PMID- 15750115 TI - Genetic diversity of multidrug-resistant Mycobacterium tuberculosis isolates and identification of 11 novel rpoB alleles in Taiwan. AB - Of 162 multidrug-resistant Mycobacterium tuberculosis isolates from Taiwan, 60.5% were found to belong to the Beijing family on the basis of spoligotyping results. IS6110 restriction fragment length polymorphism fingerprinting showed genetic diversity among the multidrug-resistant isolates. Furthermore, 90.1% of the multidrug-resistant isolates had mutations in the rpoB gene, and 11 novel alleles were recognized. PMID- 15750116 TI - Diminished replicative fitness of primary human immunodeficiency virus type 1 isolates harboring the K65R mutation. AB - The human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) resistance mutation K65R confers intermediate levels of resistance to several RT inhibitors, including a three- to fourfold reduction of tenofovir susceptibility. Here, we have used for the first time primary HIV-1 isolates from individuals who developed the K65R mutation while enrolled in a clinical trial of tenofovir to analyze the impact of this mutation on HIV-1 replicative fitness. A marked impairment in replicative fitness was observed in association with the selection of viruses carrying the K65R mutation in all patients. The mean replicative fitness among these viruses was 20% relative to the corresponding baseline wild type virus, ranging from 10 to 32% depending on the accompanying RT mutations. These results support a reduction in in vivo replication for K65R mutant viruses. PMID- 15750117 TI - Genomic relatedness of the new Matlab variants of Vibrio cholerae O1 to the classical and El Tor biotypes as determined by pulsed-field gel electrophoresis. AB - The genomes of the recently described Matlab variants of Vibrio cholerae O1 that are hybrids between classical and El Tor biotypes were compared with those of El Tor and classical biotypes by the use of pulsed-field gel electrophoresis. Dendrograms constructed using the unweighted-pair group method using average linkages generated from NotI restriction patterns of whole-chromosomal DNA grouped these strains into two major clusters that were found to be similar but not identical to those of either of the biotypes. Strains that clustered with the classical biotype appear to have been derived from the classical strains, which are thought to be extinct. PMID- 15750118 TI - Infective endocarditis caused by Granulicatella elegans originating in the oral cavity. AB - We studied the pheno- and genotypes of an oral Granulicatella elegans strain in comparison with those of a blood-derived isolate which caused infective endocarditis. The two isolates exhibited identical biochemical characteristics and had the same drug MICs. Their genotypes were indistinguishable, indicating that these were from the same clone. The transmission of G. elegans from the oral cavity thus should be noted as a possible cause of infective endocarditis. PMID- 15750119 TI - Presence of DNA of human papillomavirus 16 but no other types in tumor-free tonsillar tissue. AB - According to PCR, the prevalences of human papillomavirus (HPV) DNA were 6.3% (13 of 206) in tonsillitis or hypertrophic tonsillar tissues and 0.6% (1 of 174) in exfoliated cells from normal tonsils. HPV-16 was the only type detected in tonsillar tissues, but it did not appear to lead to L1 antibody production. PMID- 15750120 TI - Detection of human metapneumovirus RNA sequences in nasopharyngeal aspirates of young French children with acute bronchiolitis by real-time reverse transcriptase PCR and phylogenetic analysis. AB - Human metapneumovirus (HMPV) was the unique viral pathogen detected by a real time reverse transcriptase PCR (RT-PCR) assay in 6 (6.4%) of 94 consecutive French children hospitalized for acute bronchiolitis from September 2001 to June 2002. This virus was identified as the third etiological cause of bronchiolitis, after respiratory syncytial virus and rhinovirus (35 [37%] and 21 [22%] of 94 cases, respectively). Phylogenetic analysis of F-gene sequences demonstrated the cocirculation of distinct HMPV genotypes during this study. These findings highlight the need to implement a rapid HMPV RT-PCR detection assay for the clinical diagnosis of respiratory infections in pediatric patients with bronchiolitis. PMID- 15750121 TI - The pls gene found in methicillin-resistant Staphylococcus aureus strains is common in clinical isolates of Staphylococcus sciuri. AB - pls, a gene found in type I staphylococcal cassette chromosome mec (SCCmec) regions of methicillin-resistant Staphylococcus aureus strains, was present in 12 of the 15 human clinical Staphylococcus sciuri isolates studied. Pls was expressed in the S. sciuri isolates, although at a lower level than in S. aureus. Other parts of SCCmec could also be found in the S. sciuri genome. PMID- 15750122 TI - Genetic variability among serotype G4 Italian human rotaviruses. AB - A total of 254 serotype GH rotavirus strains were detected in Palermo, Italy, from 1985 to 2003. Out of 38 serotype G4 strains selected for genetic analysis, 14 were recognized by genotyping as type G9. Strains confirmed to belong to the G4 type showed temporal patterns of genetic evolution in their VP7 and VP4 gene sequences, and the latest Italian G4 strains were distantly related to the reference vaccinal ST3 strain. PMID- 15750123 TI - Sugars inhibit expression of the rugose phenotype of Vibrio cholerae. AB - Vibrio cholerae can shift to a rugose colony phenotype, reflecting expression of an exopolysaccharide that provides protection against a variety of environmental stresses. Our data indicate that expression of the rugose phenotype is inhibited by a variety of sugars, including sucrose, dextrose, arabinose, fructose, and maltose. Inhibition by sucrose may be one factor in explaining the failure of rugose strains to grow on thiosulfate citrate bile salts sucrose agar, the primary selective medium for V. cholerae. PMID- 15750124 TI - Differentiation between atypical isolates of Candida lusitaniae and Candida pulcherrima by determination of mating type. AB - We report on five clinical isolates routinely identified as Candida lusitaniae that the ID 32C system was unable to discriminate from the closely related species Candida pulcherrima. When additional tests did not allow accurate identification, the less usual mating type test identified all of them as Clavispora lusitaniae. Mating type testing appears to be a valuable tool for assessing the true incidence of this emerging non-albicans Candida species. PMID- 15750125 TI - Survey of emm-like gene sequences from pharyngeal isolates of group C and group G streptococci collected in Spain. AB - We conducted a nationwide surveillance of the variable 5' emm-like (M-like protein gene) sequences from 214 pharyngeal group C and group G streptococci. Almost 75% of the isolates exhibited emm or emm-like sequences previously described. We identified six new 5' emm-like regions, and almost 23% of the isolates were nontypeable. Five emm-like sequences accounted for more than 50% of the isolates in group C and group G, suggesting horizontal gene transfer between strains of different species. PMID- 15750126 TI - Molecular epidemiology of meningococci isolated in Niger in 2003 shows serogroup A sequence type (ST)-7 and serogroup W135 ST-11 or ST-2881 strains. AB - In 2003, in the Zinder and Maradi regions (Niger), epidemics were due to serogroup A:4:P1.9 meningococci belonging to sequence type 7 (ST-7). In Niamey, only sporadic cases were reported: 55% of the meningococcus strains were in serogroup A, and 38% were in serogroup W135 and could be placed in ST-11, identical to the 2002 Burkina Faso epidemic clone, and in ST-2881, a new ST. PMID- 15750127 TI - Rate and incidence estimates of recent human immunodeficiency virus type 1 infections among pregnant women in Sao Paulo, Brazil, from 1991 to 2002. AB - The serological testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) was employed to estimate HIV incidence among pregnant women from Sao Paulo, Brazil. A cross-sectional study (1999 to 2002) showed an incidence of infection of 0.2 per 100 pregnant women per year (95% confidence interval, 0.041 to 0.608). Western blot profiles suggested an association between results of the STARHS analysis and gp41/gp31 bands. PMID- 15750128 TI - Dihydropteroate synthase and novel dihydrofolate reductase gene mutations in strains of Pneumocystis jirovecii from South Africa. AB - Dihydropteroate synthase (DHPS) gene mutations have raised concerns about emerging sulfonamide resistance in Pneumocystis jirovecii. DHPS and dihydrofolate reductase (DHFR) gene products were amplified in clinical specimens from South African patients. One of 53 DHPS genes sequenced contained the double mutation Thr55Ala Pro57Ser. DHFR gene mutations detected were Ala67Val and the new mutations Arg59Gly and C278T. PMID- 15750129 TI - Comparison of COBAS AMPLICOR Neisseria gonorrhoeae PCR, including confirmation with N. gonorrhoeae-specific 16S rRNA PCR, with traditional culture. AB - A total of 3,023 clinical specimens were tested for Neisseria gonorrhoeae by using COBAS AMPLICOR (CA) PCR and confirmation of positives by N. gonorrhoeae specific 16S rRNA PCR. The sensitivity of CA plus 16S rRNA PCR was 98.8%, compared to 68.2% for culture. Confirmation of CA positives increased the positive predictive value from 54.8 to 96.6%. PMID- 15750130 TI - Erroneous reporting of coagulase-negative Staphylococci as Kocuria spp. by the Vitek 2 system. AB - Misidentification of coagulase-negative staphylococci (CoNS) may delay appropriate treatment. We investigated 20 clinical isolates identified as Kocuria spp. by the Vitek 2 system. All were identified as CoNS by 16S rRNA gene sequencing (18 Staphylococcus epidermidis, 1 Staphylococcus haemolyticus). Four Kocuria isolates were shown to be identical to CoNS from the same patient by pulsed-field gel electrophoresis. Isolates identified by Vitek 2 as Kocuria most likely represent misidentified CoNS, and if clinically indicated, should be investigated further by genomic methods. PMID- 15750131 TI - Arthrobacter scleromae sp. nov. isolated from human clinical specimens. AB - A gram-positive, coryneform bacterium was isolated from swollen scleromata of a dermatosis patient. An analysis of its phenotypic, chemotaxonomic, and genotypic characteristics showed that this bacterium is closely associated with Arthrobacter oxydans and Arthrobacter polychromogenes but that it belongs to a distinct species, for which the name Arthrobacter scleromae sp. nov. is proposed. PMID- 15750132 TI - False-positive Histoplasma capsulatum Gen-Probe chemiluminescent test result caused by a Chrysosporium species. AB - We describe a case in which the Histoplasma capsulatum AccuProbe test displayed cross-reactivity with a respiratory isolate thought to be Histoplasma but not morphologically consistent with H. capsulatum. The isolate was later identified as the Chrysosporium anamorph of Nannizziopsis vriesii by sequence analysis and phenotypic data. PMID- 15750133 TI - Osteomyelitis caused by Enterobacter cancerogenus infection following a traumatic injury: case report and review of the literature. AB - We report a case of osteomyelitis caused by Enterobacter cancerogenus resistant to aminopenicillins in a 56-year-old male who had a motorcycle accident and suffered from multiple bone fractures with abundant environmental exposure. E. cancerogenus has rarely been associated with human infections, and its clinical significance remains unclear. PMID- 15750134 TI - Adenovirus type F subtype 41 causing disseminated disease following bone marrow transplantation for immunodeficiency. AB - Adenovirus causes disseminated disease following bone marrow transplantation (BMT). We report a child who underwent T-cell-depleted BMT. Adenovirus subgenus F serotype 41 was detected antemortem by PCR in cerebrospinal fluid and postmortem in other tissues. Serotypes 40 and 41, associated with gastrointestinal disease, have not previously been implicated in disseminated disease. PMID- 15750135 TI - Spondylitis due to Mycobacterium xenopi in a human immunodeficiency virus type 1 infected patient: case report and review of the literature. AB - Nontuberculous mycobacterial infections are well known to occur in patients with human immunodeficiency virus infection. However, spondylitis due to mycobacteria other than Mycobacterium tuberculosis is uncommon. We report a case of biopsy- and culture-proven Mycobacterium xenopi spondylitis in an AIDS patient and discuss approaches to diagnosis and therapy. This case serves to highlight the potential pathogenic role of this usually environmental commensal organism in severely immunosuppressed AIDS patients and uncertainties in their management, given the scarce data on appropriate therapy for this organism. PMID- 15750137 TI - Typhoid fever associated with acute appendicitis caused by an H1-j strain of Salmonella enterica serotype Typhi. AB - While most strains of Salmonella enterica serotype Typhi, the etiologic agent of typhoid fever, have only a phase 1 flagellar antigen, H1-d, variations of the flagellar antigen have been observed. Although H1-j strains (one of the flagellar antigen variants) account for 10 to 50% of S. enterica serotype Typhi strains found in Indonesia, there have been no published data to suggest its existence in other parts of the world. We describe a case of typhoid fever associated with acute appendicitis caused by an S. enterica serotype Typhi H1-j strain in a Chinese woman in Hong Kong. A gram-negative, motile rod was recovered from her blood and stool cultures. Conventional biochemical tests and the Vitek system (GNI+) showed that the bacterium was S. enterica serotype Typhi. The isolate agglutinated with poly(O), 9O, Vi and H1-j Salmonella antisera but not with poly(H) antisera. The patient developed antibodies against only S. enterica serotype Typhi O antigens but not against H1-d antigen by the Widal test. Flagellin C gene (fliC) sequencing showed a 261-bp deletion in the fliC gene of the isolate, confirming that the isolate possessed the H1-j antigen. The patient had no past history of travel to Indonesia or personal contact with any Indonesian. She recovered with appendectomy and antibiotic treatment. Further studies should be performed to determine the prevalence of this unusual S. enterica serotype Typhi strain in our locality. PMID- 15750138 TI - Pulmonary Mycobacterium intermedium disease in an elderly man with healed pulmonary tuberculosis. AB - A 76-year-old man with a history of pulmonary tuberculosis was found to be sputum smear positive for acid-fast bacilli. The 16S rRNA sequence identified the culture isolate as Mycobacterium intermedium, the pathogenicity of which has not been confirmed. Chemotherapy with isoniazid, rifampin, and ethambutol resulted in clinical improvement. PMID- 15750136 TI - First isolation of Bacteroides thetaiotaomicron from a patient with a cholesteatoma and experiencing meningitis. AB - A 45-year-old man with a cholesteatoma experienced purulent meningitis. Microbial analysis of cerebrospinal fluid yielded in pure culture a gram-negative bacillus. Phenotypic methods were suggestive of a Bacteroides distasonis or either a Bacteroides thetaiotaomicron or Bacteroides ovatus infection. The isolate was identified by 16S rRNA gene sequence analysis as B. thetaiotaomicron. This is the first case of B. thetaiotaomicron meningitis in pure culture. PMID- 15750139 TI - Endophthalmitis caused by Enterococcus mundtii. AB - Enterococcus mundtii has rarely been isolated from environmental or human sources. We report the identification of E. mundtii as a pathogen of human infectious disease by DNA sequencing of 16S rRNA and sodA genes in a case of endophthalmitis developed in a 66-year-old immunocompetent gardener. PMID- 15750140 TI - Mixed bacterial meningitis due to Streptococcus pneumoniae and Neisseria meningitidis in an 18-month-old child. AB - We report an unusual case of culture-proven pneumococcal and meningococcal mixed meningitis in an 18-month-old girl. The patient responded well to antimicrobial therapy and recovered completely without sequelae. No underlying condition could be demonstrated except a rhinitis of unknown etiology 2 days before the onset of the symptoms suggesting meningitis. PMID- 15750141 TI - Fatal peritonitis caused by Pasteurella multocida capsular type F in calves. AB - A fatal case of atypical septicemia of pasteurellosis in veal calves is described. The causative organism was identified as a multiresistant Pasteurella multocida capsular type F isolate. The outbreak was characterized by fibrinous peritonitis and mortality, which are hitherto unreported features of P. multocida capsular type F infections. PMID- 15750142 TI - Airway colonization by Acrophialophora fusispora in patients with cystic fibrosis. AB - Acrophialophora fusispora is a thermotolerant soil fungus which is very unusual in clinical samples. Here we report four cases of transient or chronic airway colonization by A. fusispora in patients with cystic fibrosis (CF). However, the prevalence of this fungus in CF patients may be underestimated due to the currently poor knowledge of this fungus in most of the medical mycology laboratories. In addition, its clinical importance regarding CF remains to be evaluated. PMID- 15750143 TI - Screening for extended-spectrum beta-lactamase-producing pathogenic enterobacteria in district general hospitals. PMID- 15750144 TI - Association between cerebrospinal fluid pleocytosis and enteroviral meningitis. PMID- 15750145 TI - Amplification of coccidioidal DNA in clinical specimens by PCR. PMID- 15750146 TI - Radioligands for brain imaging of the kappa-opioid system. PMID- 15750147 TI - Is the "ideal" gamma-probe for intraoperative radioguided surgery conceivable? PMID- 15750148 TI - The direct route may not be the best way to home. PMID- 15750149 TI - Prospective comparison of 3 gamma-probes for sentinel lymph node detection in 200 breast cancer patients. AB - Previous reports have shown that axillary sentinel lymph node (ASLN) radiodetection allows accurate axillary staging for patients with early breast cancer. Radioguided surgery implies the use of a gamma-probe to count the emitted radioactivity of marked ASLNs. Several gamma-probes are commercially available, each with its own properties. The clinical impact of the type of gamma-probe used for ASLN radiodetection remains to be evaluated. METHODS: Three commercially available gamma-probes were evaluated: a scintillator with a bismuth germanate crystal (probe A), a semiconductor with a cadmium telluride crystal (probe B), and a semiconductor with a cadmium zinc telluride crystal (probe C). Two hundred patients with early breast cancer were prospectively enrolled to undergo ASLN radiodetection and axillary lymphadenectomy. ASLN mapping consisted of injecting (99m)Tc-sulfur-colloid around the tumor. For each patient, sentinel lymph nodes were counted successively with the 3 probes and the sensitivity of each gamma probe was determined from ASLN residual activity. The results of detection rates and false-negative rates for each probe were compared. RESULTS: Mean residual ASLN activity was 52 kBq (range, 0.07-189 kBq). Sensitivity was compared among the 3 probes and found to be best for probe A. The detection rate of probe A was significantly better than that of probe B (93% vs. 86%, P = 0.05) but not different from that of probe C (93% vs. 90%). No differences in false-negative rates were observed among the 3 probes. CONCLUSION: ASLN detection rate depends on the type of gamma-probe used. Because failure to detect the ASLN leads to complete axillary lymphadenectomy, involving local morbidity and other sequelae, the type of gamma-probe must be considered important for sentinel lymph node radiodetection. PMID- 15750150 TI - Comparison of 18F-FLT PET and 18F-FDG PET in esophageal cancer. AB - 18F-FDG PET has gained acceptance for staging of esophageal cancer. However, FDG is not tumor specific and false-positive results may occur by accumulation of FDG in benign tissue. The tracer 18F-fluoro-3'-deoxy-3'-L-fluorothymidine (18F-FLT) might not have these drawbacks. The aim of this study was to investigate the feasibility of 18F-FLT PET for the detection and staging of esophageal cancer and to compare 18F-FLT PET with 18F-FDG PET. Furthermore, the correlation between 18F FLT and 18F-FDG uptake and proliferation of the tumor was investigated. METHODS: Ten patients with biopsy-proven cancer of the esophagus or gastroesophageal junction were staged with CT, endoscopic ultrasonography, and ultrasound of the neck. In addition, all patients underwent a whole-body 18F-FLT PET and 18F-FDG PET. Standardized uptake values were compared with proliferation expressed by Ki 67 positivity. RESULTS: 18F-FDG PET was able to detect all esophageal cancers, whereas 18F-FLT PET visualized the tumor in 8 of 10 patients. Both 18F-FDG PET and 18F-FLT PET detected lymph node metastases in 2 of 8 patients. 18F-FDG PET detected 1 cervical lymph node that was missed on 18F-FLT PET, whereas 18F-FDG PET showed uptake in benign lesions in 2 patients. The uptake of 18F-FDG (median standardized uptake value [SUV(mean)], 6.0) was significantly higher than 18F-FLT (median SUV(mean), 3.4). Neither 18F-FDG maximum SUV (SUV(max)) nor 18F-FLT SUV(max) correlated with Ki-67 expression in the linear regression analysis. CONCLUSION: In this study, uptake of 18F-FDG in esophageal cancer is significantly higher compared with 18F-FLT uptake. 18F-FLT scans show more false negative findings and fewer false-positive findings than do 18F-FDG scans. Uptake of 18F-FDG or 18F-FLT did not correlate with proliferation. PMID- 15750151 TI - The value of 18F-FDG PET in the detection of stage M0 carcinoma of the nasopharynx. AB - Distant metastasis is an important issue for nasopharyngeal carcinoma (NPC). The potential value of PET using 18F-FDG has not been well defined. This prospective study investigated the impact of 18F-FDG PET in NPC patients with stage M0 disease. METHODS: From April 2001 to June 2003, 140 NPC patients (118 primary and 22 primary recurrent) with stage M0 (negative results from chest radiography, liver sonography, and whole-body bone scanning) underwent 18F-FDG PET to check for distant metastases. Confirmatory MRI or CT was performed if any abnormal 18F FDG uptake was found at distant sites. The distant lesion was confirmed pathologically, if feasible, and was followed up clinically and with imaging for at least 6 mo. RESULTS: 18F-FDG PET detected 26 true-positive metastatic sites in 18 (12.9%) of the 140 patients, among whom 14 had primary and 4 had recurrent tumors. The patient-based sensitivity and specificity of 18F-FDG PET for distant metastases were 100% and 86.9%, respectively. Mediastinal lymph nodes (n = 8) were the most common sites, followed by lung, liver, and bone (n = 5 each) and by other lymph nodes (n = 3). In patients with primary tumors, advanced nodal status (N2-3) was a statistically significant variable associated with development of distant metastases (P = 0.044). For recurrent NPC, neither age, sex, initial tumor stage, grade of differentiation, nor nodal stage showed a statistically significant difference between patients with and patients without distant metastases. CONCLUSION: 18F-FDG PET is valuable in avoiding aggressive locoregional radiotherapy in some NPC patients by the revelation of occult distant metastases, especially in patients with primary disease at a nodal stage of N2-3. PMID- 15750152 TI - PET with O-(2-18F-Fluoroethyl)-L-Tyrosine in peripheral tumors: first clinical results. AB - O-(2-18F-Fluoroethyl)-L-Tyrosine (18F-FET) PET has shown promising results in brain tumor diagnosis. The aim of this prospective study was to evaluate 18F-FET PET in comparison with 18F-FDG PET in patients with peripheral tumors. METHODS: Forty-four consecutive patients with suspected malignant tumors underwent 18F-FET PET and 18F-FDG PET within 7 d. Whole-body PET studies were performed 1 h after intravenous injection of 370 MBq of 18F-FET or 18F-FDG. Six patients were excluded from the analysis because a malignant tumor could not be verified. In 38 patients (7 with colorectal cancer, 6 with pancreatic cancer, 9 with head-neck cancer, 4 with lymphomas, 3 with lung cancer, 3 with ovarian cancer, 4 with breast cancer, and 2 with prostatic cancer), 18F-FET PET and 18F-FDG PET were compared. RESULTS: 18F-FET was positive in only 13 of 38 patients (8 with head neck cancer, 3 with breast cancer, and 2 with lung cancer), whereas 18F-FDG exhibited increased uptake in 37 of 38 patients. All squamous cell carcinomas were found to be 18F-FET-positive tumors (8 head-neck cancer and 2 lung cancer), whereas most adenocarcinomas were found to be 18F-FET-negative tumors. In patients with colorectal cancer, pancreatic cancer, ovarian cancer, prostatic cancer, and lymphomas, no increased 18F-FET uptake could be identified. All lesions that exhibited increased 18F-FET uptake also showed increased 18F-FDG uptake. No additional lesion was identified by 18F-FET PET but not by 18F-FDG PET. A subgroup analysis of patients with head-neck carcinomas allowed a better distinction between malignant and inflammatory tissues with 18F-FET than with 18F FDG. CONCLUSION: 18F-FET is inferior to 18F-FDG as a PET tracer for general tumor diagnosis. Our preliminary results suggest rather selective uptake of 18F-FET in squamous cell carcinomas. Compared with 18F-FDG PET, 18F-FET PET may allow a better distinction between tumors and inflammatory tissues in patients with squamous cell carcinomas. PMID- 15750153 TI - Prognostic value of myocardial ischemia in patients with uncomplicated acute myocardial infarction: direct comparison of stress echocardiography and myocardial perfusion imaging. AB - This study directly compared the prognostic value of predischarge dobutamine stress echocardiography (DSE) and dobutamine myocardial SPECT perfusion imaging in patients with prior myocardial infarction. METHODS: The study population consisted of 146 consecutive patients who underwent predischarge DSE and SPECT with (99m)Tc-sestamibi after a first acute uncomplicated myocardial infarction treated with thrombolysis. Fifty patients who underwent revascularization within 90 d from the imaging studies were excluded. Cardiac death and nonfatal myocardial infarction were considered events. Follow-up was 98% complete in a mean period of 44 +/- 19 mo. RESULTS: Myocardial ischemia was detectable in 55 (58%) patients at SPECT and in 63 (67%) patients at DSE. Concordance between SPECT and DSE in detecting ischemia was observed in 68 (72%) of the 94 patients (kappa value, 0.41). During the follow-up, there were 20 cardiac events (9 deaths and 11 nonfatal myocardial infarctions). Ischemia at SPECT was a significant predictor of events (hazards ratio = 4.8; 95% confidence interval, 1.4-16.3; P < 0.01). However, ischemia at DSE (biphasic or worsening patterns) was not associated with events, whereas biphasic pattern alone was associated with a poor outcome compared with direct worsening (P < 0.05). Finally, at Cox multivariate analysis, ischemia at SPECT but not biphasic pattern at DSE was a significant independent predictor of events (P < 0.01). CONCLUSION: These results indicate that, after uncomplicated myocardial infarction, ischemia at SPECT is associated with an increased risk of cardiac events at long-term follow-up. However, ischemia at DSE was unable to stratify patients after myocardial infarction. PMID- 15750154 TI - Influence of reconstruction iterations on 18F-FDG PET/CT standardized uptake values. AB - The goal of this study was to evaluate the effect on the average standardized uptake value (avgSUV) and maximum standardized uptake value (maxSUV) of changing the number of iterations in the reconstruction process on studies acquired with PET/CT. METHODS: Data from 50 human tumors were acquired on a PET/CT scanner, using the CT portion for attenuation correction. Reconstruction was performed using the 2-dimensional reconstruction method of ordered-subsets expectation maximization (OSEM) with 28 subsets and with 1, 2, 3, 4, 5, 10, 20, and 40 iterations. The standardized uptake value (SUV) of the studies was analyzed by positioning a region of interest tightly around the tumor and reproducing the same area on all same-study iterations for SUV measurements. RESULTS: The differences in mean avgSUV and mean maxSUV were statistically different across different iteration groups. SUV data demonstrated that the avgSUV measurements have the most significant differences between 1 versus 2 iterations and 2 versus 3 iterations. The P values for these comparisons were less then 0.001. For maxSUV, all differences had P values less than 0.001. There also was a systematic increase in the SUVs as the number of iterations increased. The avgSUV increased at early iterations (less than 5), with just 50%-60% increasing after 5 iterations. However, maxSUV increased systematically at early iterations, and this trend continued as the number of iterations increased. CONCLUSION: The OSEM algorithm converges sooner for avgSUV than for maxSUV. The likely reason is that avgSUV depends on low-frequency features that are recovered with fewer iterations. The differences in maxSUV were likely due to noise, which increased with the number of iterative updates, and to increased resolution and recovery of high-frequency features (i.e., tumor heterogeneity) with a larger number of iterations. Factors that determine the quantitative accuracy of iterative reconstruction may have played an additional role. Given the continued change in maxSUV with iterations, great care must be taken in selecting the number of iterative updates when using it to assess tumors and their response to chemotherapy and radiation therapy. Because 2-5 iterations with 8-28 subsets are being used in clinical settings, these data are pertinent when comparing the SUVs of a tumor before and after therapy. PMID- 15750155 TI - Optimized intravenous contrast administration for diagnostic whole-body 18F-FDG PET/CT. AB - Standard application of CT intravenous contrast agents in combined PET/CT may lead to high-density artifacts on CT and attenuation-corrected PET. To avoid associated diagnostic pitfalls, we designed and compared different intravenous contrast injection protocols for routine whole-body PET/CT. METHODS: Whole-body PET/CT included a topogram and a single spiral CT scan (2-row) with or without intravenous contrast, followed by an emission scan. The CT scan was used for attenuation correction of the emission data. Four groups of 10 whole-body PET/CT referrals each were investigated: (A) no intravenous contrast agent, (B) biphasic injection (90 and 50 mL at 3 and 1.5 mL/s, respectively) of intravenous contrast (300 mg/mL iodine) and CT in the craniocaudal direction with a 30-s delay, (C) triple-phase injection (90, 40, and 40 mL at 3, 2, and 1.5 mL/s, respectively) in the craniocaudal direction with a 50-s delay, and (D) dual-phase injection (80 and 60 mL at 3 and 1.5 mL/s, respectively) in the caudocranial direction with a 50-s delay. CT image quality was assessed on a scale from 1 to 3, and CT and attenuation-corrected PET images were reviewed separately for contrast-induced artifacts. RESULTS: Average CT image quality was poorest for protocol A (1.0) but improved to 2.8 when using intravenous contrast agents (protocols B-D). Only protocols B and C resulted in contrast-induced image artifacts that were limited to the thorax. The most homogeneous intravenous contrast enhancement without high density image artifacts on either CT or PET after CT-based attenuation correction was achieved with protocol D. CONCLUSION: Dual-phase intravenous contrast injection and CT in the caudocranial direction with a 50-s delay yields reproducible high image quality and is now used routinely for combined diagnostic PET/CT at our hospital. PMID- 15750156 TI - Do implanted pacemaker leads and ICD leads cause metal-related artifact in cardiac PET/CT? AB - Artifacts related to metallic implants are an established limitation of CT-based attenuation correction (CT-AC) in PET/CT. However, the impact of metallic components of pacemaker leads and implantable cardioverter defibrillator (ICD) leads on the accuracy of cardiac PET has not been evaluated. The goal of this study was to investigate the magnitude of artifacts related to pacing and defibrillation leads in both phantom and patient studies. METHODS: Images were acquired on a PET/CT scanner using CT-AC and were compared with those obtained on a dedicated PET scanner using transmission source-based attenuation correction. Phantoms consisting of pacemaker leads and ICD leads submerged in uniform background activity solution were imaged, and regions were analyzed to measure radionuclide concentrations at known lead locations relative to background. In addition, 15 cardiac 18F-FDG patients (having either pacing leads, defibrillation leads, or both) were imaged on both PET/CT and PET scanners. Images were visually and quantitatively assessed to determine whether artifact related to the implanted leads was present and, if so, its severity relative to surrounding myocardium. RESULTS: In phantom studies, artifacts caused by pacing lead electrodes were barely noticeable, but artifacts arising from highly radioopaque ICD shock coil electrodes were clearly apparent. In the patient studies, no artifacts from pacing leads were identified. However, significant artifact was observed in 50% of the patient studies with ICD leads. In the affected areas, local myocardial uptake in PET/CT images using CT-AC was, on average, 30% higher than that in the corresponding PET images. CONCLUSION: Although pacemaker leads do not appear to cause artifact in cardiac PET/CT images, ICD leads frequently do result in artifacts of sufficient magnitude to impact clinical image interpretation. Accordingly, software-based corrections in CT-AC algorithms appear necessary for accurate cardiac imaging with PET/CT. PMID- 15750157 TI - The diabetic foot: initial experience with 18F-FDG PET/CT. AB - Osteomyelitis complicates up to one third of diabetic foot infections, is often due to direct contamination from a soft-tissue lesion, and represents a clinical challenge. Early diagnosis is important since antibiotic therapy can be curative and may prevent amputation. The present study assessed the role of PET/CT using 18F-FDG for the diagnosis of diabetic foot osteomyelitis. METHODS: Fourteen diabetic patients (10 men and 4 women; age range, 29-70 y) with 18 clinically suspected sites of infection underwent PET/CT after the injection of 185-370 MBq of 18F-FDG for suspected osteomyelitis complicating diabetic foot disease. PET, CT, and hybrid images were independently evaluated for the diagnosis and localization of an infectious process. Additional data provided by PET/CT for localization of infection in the bone or soft tissues were recorded. The final diagnosis was based on histopathologic findings and bacteriologic assays obtained at surgery or at clinical and imaging follow-up. RESULTS: PET detected 14 foci of increased 18F-FDG uptake suspected as infection in 10 patients. PET/CT correctly localized 8 foci in 4 patients to bone, indicating osteomyelitis. PET/CT correctly excluded osteomyelitis in 5 foci in 5 patients, with the abnormal 18F FDG uptake limited to infected soft tissues only. One site of mildly increased focal 18F-FDG uptake was localized by PET/CT to diabetic osteoarthropathy changes demonstrated on CT. Four patients showed no abnormally increased 18F-FDG uptake and no further evidence of an infectious process on clinical and imaging follow up. CONCLUSION: 18F-FDG PET can be used for diagnosis of diabetes-related infection. The precise anatomic localization of increased 18F-FDG uptake provided by PET/CT enables accurate differentiation between osteomyelitis and soft-tissue infection. PMID- 15750158 TI - 18F-CPFPX PET identifies changes in cerebral A1 adenosine receptor density caused by glioma invasion. AB - Adenosine plays a critical role in both tumor proliferation and the cerebral response to tumor invasion. We used 8-cyclopentyl-3-(3-18F-fluoropropyl)-1 propylxanthine (18F-CPFPX) PET to investigate A1 adenosine receptor (A1AR) density as a potential indicator of the local cerebral response to glioma invasion. METHODS: A1AR density in F98 glioma-bearing rats was examined by 18F CPFPX and 3H-CPFPX using PET, quantitative in vitro and ex vivo double-label receptor autoradiography, and immunohistochemical analyses. RESULTS: For all imaging modalities, A1AR signal intensity was increased in a zone surrounding experimental tumors (136%-146% that in control tissue) (P < 0.01). Immunostaining identified activated astrocytes as the main origin of peritumoral A1AR upregulation. The results of a pilot 18F-CPFPX PET study on a patient with recurrent glioblastoma multiforme confirmed increases in A1AR density in the immediate vicinity of the tumor. CONCLUSION: 18F-CPFPX PET is suitable for the detection of peritumoral changes in A1AR density. Molecular imaging with 18F CPFPX PET may open novel possibilities for gaining experimental and clinical insights into the cerebral response to tumor invasion. PMID- 15750159 TI - Performance evaluation of the microPET focus: a third-generation microPET scanner dedicated to animal imaging. AB - The microPET Focus is the latest generation microPET system dedicated to high resolution animal imaging and incorporates several changes to enhance its performance. This study evaluated the basic performance of the scanner and compared it with the Primate (P4) and Rodent (R4) models. METHODS: The system consists of 168 lutetium oxyorthosilicate (LSO) detectors arranged in 4 contiguous rings, with a 25.8-cm diameter and a 7.6-cm axial length. Each detector consists of a 12 x 12 LSO crystal array of 1.51 x 1.51 x 10.00 mm3 elements. The scintillation light is transmitted to position-sensitive photomultiplier tubes via optical fiber bundles. The system was evaluated for its energy and spatial resolutions, sensitivity, and noise equivalent counting rate. Phantoms and animals of varying sizes were scanned to evaluate its imaging capability. RESULTS: The energy resolution averages 18.5% for the entire system. Reconstructed image resolution is 1.3-mm full width at half maximum (FWHM) at the center of field of view (CFOV) and remains under 2 mm FWHM within the central 5 cm-diameter FOV in all 3 dimensions. The absolute sensitivity of the system is 3.4% at the CFOV for an energy window of 250-750 keV and a timing window of 10 ns. The noise equivalent counting-rate performance reaches 645 kcps for a mouse size phantom using 250- to 750-keV and 6-ns settings. Emission images of a micro Derenzo phantom demonstrate the improvement in image resolution compared with previous models. Animal studies exhibit the capability of the system in studying disease models using mouse, rat, and nonhuman primates. CONCLUSION: The Focus has significantly improved performance over the previous models in all areas evaluated. This system represents the state-of-the-art scintillator-based animal PET scanner currently available and is expected to advance the potential of small animal PET. PMID- 15750161 TI - Radiolabeled RGD uptake and alphav integrin expression is enhanced in ischemic murine hindlimbs. AB - Radiolabeled RGD peptides that target alpha(v)beta3 integrin are promising tracers for imaging tumor angiogenesis. Integrins and angiogenesis also play important roles in healing of ischemic lesions. Thus, we investigated the biodistribution of radiolabeled RGD and expression of alpha(v) integrin in a mouse model of hindlimb ischemia. METHODS: 125I-3-Iodo-D-Tyr4-cyclo(-Arg-Gly-Asp D-Tyr-Val-) (125I-c(RGD(I)yV)) was synthesized and tested for endothelial binding. Hindlimb ischemia was induced in ICR mice through femoral artery ablation, and perfusion was measured with laser Doppler blood flowmetry. 125I c(RGD(I)yV) biodistribution was evaluated in control animals (n = 7) and ischemic models on day 3, 8, or 14 (n = 6 each). Control experiments were performed using a radiolabeled peptide with a scrambled amino acid sequence (125I-GfVGV). Microsections of hindlimb tissue were immunostained for alpha(v) integrin expression and stained with alkaline phosphatase to localize vascular endothelial cells. RESULTS: 125I-c(RGD(I)yV) retained specific binding to human umbilical vein endothelial cells. Perfusion in ischemic hindlimbs immediately fell to 10% +/- 4% of contralateral levels and gradually recovered to 22% +/- 11% and 64% +/- 9% on days 8 and 14, respectively. 125I-c(RGD(I)yV) uptake in ischemic muscles significantly increased from a control level of 0.16 +/- 0.05 %ID/g (percentage injected dose per gram of tissue) to 0.85 +/- 0.76 %ID/g at day 3, 0.43 +/- 0.23 %ID/g at day 8, and 0.43 +/- 0.28 %ID/g at day 14 (all P < 0.05). Ischemic muscle to-lung count ratios had a virtually identical trend: 0.42 +/- 0.25 for controls, 2.34 +/- 1.70 at day 3 (P < 0.02), 1.46 +/- 0.52 at day 8 (P < 0.001), and 1.39 +/- 0.94 at day 14 (P < 0.02). In contrast, uptake of the control peptide in ischemic hindlimbs was not different from that of controls. Immunohistochemistry revealed substantially increased alpha(v) integrin staining in ischemic hindlimb tissue. CONCLUSION: Radioiodine RGD uptake is significantly enhanced in ischemic hindlimbs of a mouse model, and is accompanied by an increase in alpha(v) integrin expression. Further investigation is thus warranted to illuminate the potential role of radiolabeled RGD for noninvasive monitoring of peripheral ischemic lesions. PMID- 15750160 TI - Myelotoxicity and RBE of 211At-conjugated monoclonal antibodies compared with 99mTc-conjugated monoclonal antibodies and 60Co irradiation in nude mice. AB - The rationale of this study was to determine the myelotoxicity in nude mice of the alpha-emitter 211At conjugated to monoclonal antibodies (mAbs) and to compare the effect with an electron emitter, (99m)Tc, and external irradiation from a 60Co source, for estimation of the relative biological effectiveness (RBE). METHODS: 211At and (99m)Tc were conjugated to the IgG1 mAbs MX35 and 88BV59. Nude female BALB/c mice, 8- to 12-wk old, were injected intraperitoneally or intravenously. The biodistribution was determined 3, 6, and 18 h after injection. The bone-to-blood and bone marrow-to-blood activity concentration ratios (BBLR and BMBLR, respectively) were determined for simultaneously injected 211At- and (99m)Tc-mAbs. Bone marrow samples were taken from the femur. For each mouse, the whole-body retention was measured as well as the blood activity by repeated blood samples from the tail vein (0), 1, 3, 6, 12, and 18 h after injection. External beam irradiation from a 60Co source was also performed at 3 different dose levels. White blood cell (WBC) counts, red blood cell counts, platelet counts, and hemoglobin were determined for each mouse initially and on days 1, 4, 5, 7, 15, 22, and 27 after injection. The calculations of the absorbed dose to the bone marrow were based on the BBLR, BMBLR, the cumulated activities, and the absorbed fractions. The absorbed fractions, phi, for alpha-particles and electrons in the bone marrow were calculated using Monte Carlo simulations based on a bone marrow dosimetry model. RESULTS: The BMBLR was 0.58 +/- 0.06 and 0.56 +/- 0.06 for the 211At- and (99m)Tc-mAbs, respectively. No significant variation in BMBLR with time was found. The absorbed fractions for alpha-particles and electrons in the bone marrow were 0.88 and 0.75, respectively. The mean absorbed fractions of the photons from (99m)Tc were 0.033 and 0.52 for 140 and 18.3 keV, respectively. When different amounts of 211At- and (99m)Tc-mAbs (0.09-1.3 and 250-1,300 MBq, respectively) were administered intraperitoneally or intravenously, corresponding to absorbed doses to the bone marrow of 0.01-0.60 and 0.39-1.92 Gy, respectively, the WBC counts was suppressed by 1%-90% and 23%-89%, respectively. When external beam irradiation with a 60Co source was performed to absorbed doses of 1.4, 1.9, and 2.4 Gy, the WBC counts was suppressed by 47%-90%. These results indicate a myelotoxic in vivo RBE of 3.4 +/- 0.6 for alpha-particles compared with (99m)Tc and 5.0 +/- 0.9 compared with 60Co irradiation. CONCLUSION: The effect on the WBC counts from bone marrow irradiation with 211At-mAbs indicates an in vivo RBE of 3.4 +/- 0.6 in comparison with (99m)Tc-mAbs. The RBE value compared with external irradiation is 5.0 +/- 0.9. PMID- 15750162 TI - Development of a sodium/iodide symporter (NIS)-transgenic mouse for imaging of cardiomyocyte-specific reporter gene expression. AB - Development of a small animal imaging system for differentiated cell-specific reporter gene expression will enable us to image cellular differentiation in vivo. In this study, we developed a sodium/iodide symporter (NIS)-transgenic mouse in which NIS is constitutively expressed as an imaging reporter gene only in cardiomyocytes. METHODS: To express NIS gene in cardiomyocytes, alpha-myosin heavy chain (alpha-MHC)-NIS was constructed and used for the production of NIS transgenic mice. Twelve lines of positive founder were obtained. The adequacy of the transgenic mouse model was tested by in vivo scintigraphy, microPET, and a biodistribution study. RESULTS: The myocardium of transgenic mice showed rapid and intense uptake of 131I, which was much higher than that of the thyroid, and also showed long retention by gamma-camera pinhole imaging. The relative uptake ratio of the heart of transgenic mice was 4.6 +/- 1.5, which was 3.8 +/- 1.2 times higher than that of control wild-type mice. The uptake of the heart was completely blocked by oral administration of KClO4, an NIS inhibitor. The heart of transgenic mouse was also clearly and intensely visualized on microPET using 124I. Biodistribution data of these mice showed the uptake of 40-160 %ID/g (percentage injected dose per gram of tissue) of (99m)Tc-pertechnetate in the heart compared with 40-60 %ID/g in the stomach, respectively. NIS expression in the myocardium was confirmed by immunohistochemistry using a NIS-specific antibody. CONCLUSION: We developed a transgenic mouse model to image cardiomyocytes with a gamma-camera and microPET using an alpha-MHC promoter and NIS. The transgenic mouse can be used as an imaging model for cardiomyocyte specific reporter gene expression and cellular differentiation into cardiomyocytes after cardiac stem or progenitor cell transplantation. PMID- 15750163 TI - 11C-GR103545, a radiotracer for imaging kappa-opioid receptors in vivo with PET: synthesis and evaluation in baboons. AB - Brain kappa-opioid receptors (ORs) may be involved in several pathologic conditions, such as addiction, psychosis, and seizures. (+/-)-4-Methoxycarbonyl-2 [(1-pyrrolidinylmethyl]-1-[(3,4-dichlorophenyl)acetyl]-piperidine (GR89696) is a potent and selective kappa-OR agonist. The (-)-isomer, GR103545, is the active enantiomer of GR89696. The aim of this study was to characterize the potential of 11C-GR103545 to image kappa-OR in vivo with PET. METHODS: Brain uptake of 11C GR103545 was studied in baboons under control conditions and after blockade by naloxone (1 mg/kg intravenously). Uptake of the racemic 11C-GR89696 and of the inactive enantiomer (+)-11C-GR89696 was also evaluated. Regional total distribution volumes were derived using the arterial input function and a 2 tissue-compartment model. RESULTS: 11C-GR103545 showed excellent brain penetration and uptake kinetics, with significant washout observed within the time frame of the PET experiment. Naloxone pretreatment did not affect cerebellar total distribution volume and reduced total distribution volume in other regions to a level comparable to that in the cerebellum. The regional pattern of 11C GR103545 binding potential was consistent with the distribution of kappa-OR in primate brain, with highest levels observed in anterior cortical regions (prefrontal cortex and cingulate cortex) and striatum. In most regions, the specific-to-nonspecific equilibrium partition coefficient (V3'') ranged from 1 to 2, predicting reliable quantification. 11C-GR103545 V3'' values were approximately double the 11C-GR89696 V3'' values, whereas (+)-11C-GR89696 V3'' values were negligible, demonstrating the enantiomeric selectivity of the binding and the advantage of using the pure active enantiomer for PET studies. CONCLUSION: 11C-GR103545 is a promising PET radiotracer for imaging the kappa-OR. PMID- 15750164 TI - Pretargeting with bispecific anti-renal cell carcinoma x anti-DTPA(In) antibody in 3 RCC models. AB - We have developed an efficient pretargeting strategy for renal cell carcinoma (RCC) based on a biologically produced bispecific monoclonal antibody (bs-mAb). Tumor targeting with this 2-step pretargeting strategy in the NU-12 mouse RCC model was very efficient compared with other pretargeting strategies, possibly due to unique characteristics of the NU-12 tumor used in our studies. Here we describe the bs-mAb G250xDTIn-1 pretargeting strategy in 3 different RCC nude mouse models. METHODS: Three different human RCC xenografts in nude mice (NU-12, SK-RC-1, and SK-RC-52 tumors) were pretargeted with 100 pmol bs-mAb G250xDTIn-1. Three days after administration of the bs-mAb, mice were injected intravenously with 4 pmol 111In-labeled bivalent peptide, diDTPA-FKYK (DTPA is diethylenetriaminepentaacetic acid). At 1, 4, 24, 48, and 72 h after injection of the radiolabeled peptide, the biodistribution of the radiolabel was determined. The 3 RCC tumors were characterized in vivo and in vitro for G250 antigen expression, vessel density, vascular volume, and vascular permeability and by targeting with 111In-/125I-labeled cG250 mAb. RESULTS: Using the pretargeting strategy, relatively high uptake of the radiolabel was observed in all 3 tumor models (maximum uptake: SK-RC-1 [278 +/- 130 %ID/g (percentage injected dose per gram), 1 h after injection] > NU-12 [93 +/- 41 %ID/g, 72 h after injection] > SK RC-52 [54 +/- 9 %ID/g, 4 h after injection]). Remarkably, uptake of the radiolabel in the tumor did not correlate with G250 antigen expression. The kinetics of the radiolabel in the tumor varied largely in the 3 RCC tumors: SK-RC 1 and SK-RC-52 tumors showed a washout of the 111In label from the tumor with time: only 30% of the radiolabel was retained in the tumor 3 d after injection, whereas the 111In label was fully retained in NU-12 tumors. Physiologic characteristics (vessel density, vascular volume, and vascular permeability) of the tumors may explain these differences. CONCLUSION: G250 antigen-expressing tumors can be pretargeted very efficiently with the bs-mAb G250xDTIn-1. There was no correlation between G250 antigen expression and uptake of the radiolabel in the tumor using the pretargeting strategy or with directly labeled mAbs. Therefore, these studies show that physiologic characteristics of the tumor, such as vascular permeability, play a significant role in pretargeting. PMID- 15750165 TI - Radiation-absorbed dose from 201Tl-thallous chloride. AB - Revised radiation dosimetry estimates for 201Tl-thallous chloride have been developed using new data specifically acquired to address the issue of testicular uptake of this agent and through reevaluation of extant data for biodistribution in other organs. METHODS: Quantitative testicular scintigraphy data of sequestered testes (body-background shielded) were obtained from 28 patients (56 studies) injected with 201Tl-thallous chloride at peak exercise. Previously published data for 15 patients injected at maximal exercise were reanalyzed to obtain updated biodistribution parameters for designated organs. Radiation dose was calculated according to the MIRD schema. Radiation dose to testes as a function of age was determined. Comparisons are made between organ dose estimates derived in this study and those previously published. The dose contributions of possible contaminants (200Tl, 202Tl, 203Pb) have been included. Estimates are provided of the dose component from these contaminants if injected at the time of the maximum recommended 5-d shelf life (as opposed to at the designated calibration time). RESULTS: The radiation dose per unit administered activity to adult testes calculated in this study of 0.21 mGy/MBq (0.77 rad/mCi) is approximately a factor of 2 less than the value of 0.45 mGy/MBq (1.7 rad/mCi) previously accepted. The revised dose estimates for other organs show less variation from published values. The effective dose determined in this work is approximately 0.16 mSv/MBq (0.60 rem/mCi). Under the assumption of similar biokinetics as for the adult, the testes dose for children increases significantly as age decreases with a value of 7.5 mGy/MBq (28 rad/mCi) for a newborn. Contributions from radiocontaminants that may be encountered in the preparation of 201Tl-thallous chloride are shown to range from a fraction of a percent up to approximately 20% of the total dose for some organs, with the higher values arising from the long half-life contaminant 202Tl after a 5-d shelf life. CONCLUSION: It is recommended that the dose values determined in this study be used when estimating the radiation dose to the adult testes from intravenous administration of 201Tl-thallous chloride. The potential for increased radiation dose per administered activity to the testes at younger ages should be evaluated before performing procedures on children. The presence of radiocontaminants in the product should be considered when estimating radiation dose and may add a significant contribution to dose dependent on the specific radionuclide and concentration at the time of administration. PMID- 15750166 TI - Antisense targeting of p-glycoprotein expression in tissue culture. AB - Radiolabeled DNA and other oligomers are now under investigation for antisense targeting of a variety of messenger RNA (mRNA). Multidrug resistance (MDR) is detectable as P-glycoprotein (Pgp) expression in most cells and is often elevated in tumor cells, especially those exposed to chemotherapeutic drugs. Radiolabeled antisense DNA has not previously been considered for the targeting of mdr1 mRNA, the product of the mdr1 gene controlling Pgp expression of MDR. METHODS: A 20-mer uniform phosphorothioate DNA, described elsewhere as targeting the AUG start codon of mdr1 mRNA, was used naked along with the sense phosphorothioate DNA control. The 3 cell lines were KB-G2, an epidermal carcinoma cell line that had been transfected to overexpress mdr1 mRNA (i.e., Pgp++) compared with its parent (Pgp+) KB-31, and TCO-1, a thyroid carcinoma cell line also reported to be Pgp++. The relative expression of mdr1 mRNA in these 3 cell lines was confirmed elsewhere by reverse transcriptase polymerase chain reaction. As a marker of Pgp expression, the uptake of (99m)Tc-sestamibi was measured in the 3 cell lines after 20 h of incubation with different concentrations of both antisense and sense DNA. Both DNAs were radiolabeled with (99m)Tc via mercaptoacetyltriglycine, and cellular uptake was measured after 24 h of incubation. RESULTS: In the case of the sense DNA, the ratio of sestamibi uptake in cells incubated with the DNA to those not exposed to the DNA was unaffected regardless of cell line and regardless of DNA concentration. In contrast, this ratio was significantly higher in both the KB-G2 and TCO-1 cells when incubated with antisense DNA at concentrations greater than about 25 nmol/L (i.e., 150 ng/mL). Only in the KB-31 cells was the sestamibi accumulation unaffected by incubation with the antisense DNA. Thus, the antisense DNA was interfering with Pgp expression to a measurable extent in both Pgp++ cells, but not the Pgp+ cells. This behavior is almost certainly due to antisense targeting of mdr1 mRNA by the antisense DNA since the sense control DNA had no effect. A significant increased accumulation of (99m)Tc antisense versus (99m)Tc-sense DNA was observed in all 3 cell lines. In all cases, this difference was greatest at the lowest DNA concentrations and decreased with increasing concentration as expected for specific binding. In the KB-G2 cells, cellular accumulation of (99m)Tc-antisense DNA was strikingly high at the lowest concentration at 54%, compared with 22% for (99m)Tc-sense DNA. These accumulations therefore probably reflect the higher mRNA target concentration in the MDR++ cells than in the MDR+ cells and the higher specific binding of (99m)Tc-antisense DNA than nonspecific binding of (99m)Tc-sense DNA. CONCLUSION: Further evidence was obtained suggesting that an antisense mechanism is responsible for the accumulation of (99m)Tc-oligomers in cells in culture. Finally, whereas evidence of in vitro targeting is not necessarily evidence of in vivo targeting, our results do suggest that radiolabeled antisense DNA against the mdr1 mRNA may potentially be useful for antisense imaging of MDR in cancer. PMID- 15750167 TI - Smooth muscle cell proliferation index correlates with 111In-labeled antibody Z2D3 uptake in a transplant vasculopathy swine model. AB - Transplant vasculopathy is a major cause of morbidity and mortality in heart transplantation. The proliferation of coronary vascular smooth muscle cells is a hallmark of transplant vasculopathy. The goal of this study was to detect coronary vascular smooth muscle cell proliferation in a swine model by imaging regions of uptake of a monoclonal antibody (Z2D3) labeled with 111In. METHODS: Coronary-to-right carotid artery transplantation was performed in 10 Yucatan minipigs with coronary arteries from farm pigs as donors. In 5 of these experiments, the right carotid artery was also grafted to the left carotid artery as a homograft. In 1 farm pig, the left and right carotid arteries were switched. After 44 +/- 22 days (mean +/- SE), animals were injected with 5-bromo-2 deoxyuridine (BrDU) and 111In-Z2D3 F(ab')2. Approximately 24 h later, the pigs underwent planar and SPECT imaging. After the imaging session, the pigs were sacrificed and the vessels were removed. Ex vivo autoradiography of all grafts was performed. Next, the tissues were immersion fixed, paraffin embedded, sectioned, and stained for histologic or immunohistologic examination. Quantitative morphometry was performed. A smooth muscle cell proliferation index, calculated as (BrDU- and actin-stained cells/actin-stained cells) x 100, was correlated with in vivo and ex vivo radiotracer uptake. RESULTS: Patency or neovascularization was demonstrated in 10 of 10 allografts and 5 of 6 homografts. Ten of the scans were positive for focal tracer uptake in the neck in the area corresponding to the graft site, and 6 were negative. Actin- and BrDU-stained cells were seen in the media of allografts and in the recanalized lumen of occluded homografts. A smooth muscle cell proliferation index of 30 was used as a cutoff for scan positivity, on the basis of previous work. Analysis by the chi2 test indicated significant concordance (P < 0.01). Ex vivo vessel count ratios were significantly correlated with the smooth muscle cell proliferation index (r2 = 0.528, P < 0.01). CONCLUSION: The use of monoclonal antibody Z2D3 tagged with 111In allows the detection of proliferating smooth muscle cells and correlates with the intensity of cell proliferation. This diagnostic method could allow early noninvasive detection of transplant vasculopathy. PMID- 15750168 TI - Assessment of liver tissue after radiofrequency ablation: findings with different imaging procedures. AB - Our goal was to assess the typical appearance of normal liver tissue immediately after radiofrequency ablation (RF-ablation) when imaged with contrast-enhanced ultrasound, CT, MRI, 18F-FDG PET, and PET/CT. METHODS: Nineteen RF-ablation sessions were performed on nontumorous liver tissue of 10 Gottingen Mini Pigs. CT, ultrasound, MRI, 18F-FDG PET, and PET/CT were performed immediately after the intervention. All imaging procedures were evaluated qualitatively for areas of increased contrast enhancement (morphologic imaging) and regions of elevated tracer uptake (functional imaging). Images were assessed quantitatively by determination of ratios (r(p/p)) comparing contrast enhancement/tracer uptake in the periphery of the necrosis with contrast enhancement/tracer uptake of normal liver parenchyma. RESULTS: On morphologic imaging, an increase in contrast enhancement surrounding the ablative necrosis was detected in all lesions. Quantification of this area of increased contrast enhancement revealed ratios of r(p/p) = 1.57 +/- 0.2 for CT and r(p/p) = 1.57 +/- 0.19 for MRI. On PET and PET/CT, homogeneous tracer utilization was found surrounding all lesions. There were no areas of a focal or rim-like increase in glucose metabolism. The ratio r(p/p) was found to be 1.05 +/- 0.08 for functional data. Histologic examination revealed pooling of blood in the sinusoids of the lesion's periphery that was caused by outflow obstruction due to the central necrosis. CONCLUSION: On morphologic imaging, a rim-like increase of contrast enhancement was found immediately after RF-ablation resembling peripheral hyperperfusion. This area of contrast enhancement may hamper detection of residual tumor. On the basis of homogeneous tracer distribution surrounding the area of necrosis, PET and PET/CT may serve for early assessment of patients after RF-ablation. PMID- 15750169 TI - Dedicated pinhole SPECT of intestinal neutrophil recruitment in a mouse model of dextran sulfate sodium-induced colitis. AB - Evaluating the efficacy of therapy in experimental inflammatory bowel disease (IBD) requires information about inflammatory activity in bowel segments or leukocyte recruitment and about kinetics in the follow-up of treatment. This study evaluated a noninvasive scintigraphic technique able to assess neutrophil trafficking in a mouse model of dextran sulfate sodium (DSS)-induced colitis. METHODS: Groups of 4 BALB/c mice were assessed at baseline and after 1, 3, 5, and 8 d of treatment with DSS. Donor neutrophils were harvested by rinsing of the peritoneal cavity with phosphate-buffered saline 5 h after intraperitoneal injection of proteose peptone contained in phosphate-buffered saline and labeled with freshly prepared (99m)Tc-hexamethylpropylene amine oxime (HMPAO). Pinhole SPECT of the abdomen was performed 1 h after reinjection of 50 MBq of labeled neutrophils. In addition, the severity of inflammation was determined by histologic examination. The possibilities of the technique were illustrated by scintigraphic assessment of neutrophil trafficking with and without blocking of neutrophil migration by a CD97 monoclonal antibody in mice with DSS-induced colitis. RESULTS: Colonic uptake of (99m)Tc-HMPAO neutrophils was determined with dedicated animal pinhole SPECT in mice with DSS-induced colitis and correlated well with histologic findings (R = 0.81) and wet colon weight (R = 0.87) and moderately with clinical weight loss (R = 0.62). The neutrophil uptake ratio was reduced significantly (P < 0.01) by blocking of neutrophil migration capacity with the CD97 antibody. CONCLUSION: Animal pinhole SPECT can be used to study inflammatory activity and neutrophil recruitment in vivo in experimental colitis. PMID- 15750170 TI - In vivo characterization of 123/125I-2-iodo-L-phenylalanine in an R1M rhabdomyosarcoma athymic mouse model as a potential tumor tracer for SPECT. AB - The application of 123I-3-iodo-alpha-methyltyrosine is limited to diagnosis of brain tumors due to its marked long-term uptake in kidneys. In vitro evaluation of 125I-2-iodo-L-phenylalanine showed high uptake in R1M cells by L-type amino acid transport system 1 (LAT1). This study evaluates 123I-2-iodo-L-phenylalanine as a new specific tumor tracer for SPECT. METHODS: 123/125I-2-Iodo-L phenylalanine is prepared as a one-pot kit using the Cu1+-assisted isotopic exchange method. The characteristics of 125I-2-iodo-L-phenylalanine were examined in vivo in R1M tumor-bearing athymic mice and in acute inflammation-bearing NMRI mice. The uptake of 123/125I-2-iodo-L-phenylalanine in tumor and other organs of interest was measured by dynamic planar imaging (DPI) and gamma-counting after dissection. Displacement of 123I-2-iodo-L-phenylalanine radioactivity by L phenylalanine, L-methionine, and L-cysteine was measured. 123I-Iodo-human serum albumin planar imaging was performed to correct for blood-pool activity and MRI was performed to delineate the tumor in DPI. 18F-FDG uptake was measured with an animal PET scanner. 125I-2-Iodo-L-phenylalanine and 18F-FDG uptake in inflamed muscle were compared. RESULTS: 123/125I-2-Iodo-L-phenylalanine showed a high and fast tumor uptake and followed a reversible first-order pattern allowing calculation of the half-life and the time to reach equilibrium (t(R)). Net tumor to-background ratios up to 6.7 at 60 min were obtained. This radioactivity was significantly displaced by L-phenylalanine, L-methionine, and L-cysteine, pointing to reversible LAT transport. When plotting t(R) of the tumor uptake as a function of tumor volume, a rectangular hyperbolic curve was obtained. The almost constant t(R) values at higher tumor volumes (>4 mL) could be linked to increased necrotic tissue. Fast blood clearance of the tracer through the kidneys to the bladder and low tracer activity in the abdomen and brain were observed. The inflamed muscle showed only a slight increase of 125I-2-iodo-L-phenylalanine uptake (inflammation-to-background ratio, RIB = 1.30 +/- 0.02), in contrast to the high 18F-FDG uptake (RIB = 11.1 +/- 1.7). The in vivo stability of 123/125I-2 iodo-L-phenylalanine was good: Only 7% of free radioiodide and no other labeled metabolites were observed after 90 min. CONCLUSION: 123/125I-2-Iodo-L phenylalanine is quickly taken up by the overexpressed LAT1 system in R1M tumors with high tumor specificity. The availability of a kit and the specificity of the tracer make 123I-2-iodo-L-phenylalanine a promising tool for oncologic SPECT. PMID- 15750171 TI - Involvement of the basal ganglia in refractory epilepsy: an 18F-fluoro-L-DOPA PET study using 2 methods of analysis. AB - Studies in animal models and epileptic patients have led to the suggestion that the basal ganglia (BG) are involved in seizures. PET with 6-18F-L-3,4 fluorodihydroxyphenylalanine (18F-fluoro-L-DOPA) has recently demonstrated a reduction of striatal dopamine uptake in drug-resistant epileptic patients with ring chromosome 20 (r20) using a multiple-time graphical analysis. The aim of the present study was to evaluate the involvement of dopamine in other epileptic syndromes using a multiple-time graphical analysis and the all-brain statistical parametric mapping (SPM) analysis. METHODS: Patients with drug-resistant epilepsy were divided into 3 groups: group 1, with r20 epilepsy (n = 16; mean age +/- SD, 21.5 +/- 5.4 y); group 2, with resistant generalized "absence-like" epilepsy (n = 10; mean age, 32.3 +/- 11.4 y); and group 3, with drug-resistant temporal lobe epilepsy with hippocampal sclerosis (n = 9; mean age, 35.2 +/- 10.3 y). We compared 2 strategies of analysis of the 18F-fluoro-L-DOPA uptake constant (K(i), min(-1)) in BG using a multiple-time graphical analysis using regions of interest (the gold-standard method) and an SPM analysis using a voxel-by-voxel statistical t test to avoid a priori hypotheses in the analysis. Each epileptic group was compared with a group of healthy volunteers (n = 10; mean age, 45.1 +/- 16.5 y). RESULTS: A decrease of the mean K(i) value was observed in the striatum in all groups of patients with both types of analysis. With multiple-time graphical analysis, the reduction was evident using the averaged K(i) values over both hemispheres in each BG. Unilateral decreases in each BG were detected in SPM analysis. A ratio of decrease of 18F-fluoro-L-DOPA uptake was observed in the 3 groups of patients. Only the SPM analysis showed a decrease of 18F-fluoro-L-DOPA uptake ipsilateral to the seizure side in patients with temporal lobe epilepsy. Moreover, the all-brain SPM analysis showed a decrease of 18F-fluoro-L-DOPA uptake in the substantia nigra bilaterally (P < 0.001). CONCLUSION: This result confirms the involvement of dopamine neurotransmission in seizure control related to the type of epileptic syndrome. The difference in epileptic types may depend in part on the seizure frequency. PMID- 15750172 TI - Scintimammography with a pinhole collimator. PMID- 15750173 TI - Dedicated breast camera: is it the best option for scintimammography? PMID- 15750174 TI - Nursing home profit status and quality of care: is there any evidence of an association? AB - This article critically reviews the association between the profit status of North American nursing homes and the quality of care. Studies were identified by searching MEDLINE (January 1990-October 2002), reference lists, letters, commentaries, and editorials. The quality indicator(s) used to measure quality of care, and its relationship to profit status, was extracted from each publication. The study design and risk-adjustment methodologies used were also extracted. The interrater reliability for the extraction of these three items was determined to be 1.0, 0.6, and 0.8, respectively. A qualitative systematic review was performed using Donabedian's framework of structure, process, and outcome for analyzing medical quality of care. Empirical research in the past 12 years has found that systematic differences exist between for-profit and not-for-profit nursing homes. For-profit nursing homes appear to provide lower quality of care in many important areas of process and outcome. PMID- 15750175 TI - Does having two earners in the household matter for understanding how well employer-based health insurance works? AB - Using the 1996 Medical Expenditure Panel Survey, the authors investigate differences between households with two earners and those with a single earner in households' access to employer-based health insurance and the generosity of insurance options. They examine whether a household has an offer of coverage, whether a household holds coverage, and whether all household members are covered. They also explore whether two-earner households have more generous options as measured by the number and types of plans available, as well as contribution requirements. The authors find that having a second earner in the household dramatically improves both access to employer health insurance and the generosity of health plan choices, particularly for workers generally acknowledged to have little access, such as part-time workers and workers in small establishments. PMID- 15750176 TI - An economic evaluation of inpatient residential treatment programs in the Department of Veterans Affairs. AB - The Veterans Health Administration (VA) established psychosocial residential rehabilitation treatment programs (RTPs) to treat eligible veterans who have psychiatric and substance use disorders in a less intensive and more self-reliant inpatient setting. Forty-two (25 percent) VA medical centers adopted RTPs in 1995. Panel regression models using data from 1993 through 1999 indicated that RTPs were associated with 8.6 and 24.4 percent decreases in the average cost per day for inpatient psychiatry and substance use care, respectively. During this time, VA transitioned much of the inpatient mental health care to ambulatory services. Yet medical centers with RTPs had smaller decreases in the number of inpatient patient days than those without RTPs. Because medical centers with RTPs provided more services, this offset the per diem savings, resulting in no significant differences in total costs between medical centers with and without RPTs. PMID- 15750177 TI - How do stressful family environments relate to reported access and use of health care by low-income children? AB - This study examines the effect of stressful family environments on children's access to and use of health care, using a sample of 9,854 low-income children from the 1999 National Survey of America's Families. Indicators of stress included aspects of family structure, economic hardship, family turbulence, and parental ill health; these were combined into a composite family stress indicator. Having health insurance was the strongest predictor of health care access and use, but stressful family environments were significantly and inversely associated with parents' having confidence in the ability of family members to obtain health care, children having health care needs met, and children having any dental care in the previous year. The authors concluded that while enrollment in health insurance may be necessary to access and use health care, it is not sufficient. Stressful family environments also appear to influence the ability of parents to obtain care for their children. PMID- 15750178 TI - The impact of private and public health insurance on medication use for adults with chronic diseases. AB - This article examines the impact of public and private health insurance on the use of medications for California adults with any of four chronic diseases: heart disease, high blood pressure, diabetes, and asthma. The data set used is the 2001 California Health Interview Survey. Multivariate analyses were conducted on individuals who had been diagnosed with each of these diseases. Controlling for various demographic, health status, and employment characteristics, the authors find that the uninsured are far less likely to be taking medications for each of the conditions than those with private insurance. Interestingly, those with Medicaid coverage are even more likely than those with private insurance to be taking such medications. The results of this study underscore the importance of health insurance for all persons with chronic conditions and the benefits of Medicaid in particular for low-income adults with chronic conditions. PMID- 15750180 TI - Nuclear poly(ADP-ribose) polymerase-1 rapidly triggers mitochondrial dysfunction. AB - To obtain further information on time course and mechanisms of cell death after poly(ADP-ribose) polymerase-1 (PARP-1) hyperactivation, we used HeLa cells exposed for 1 h to the DNA alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine. This treatment activated PARP-1 and caused a rapid drop of cellular NAD(H) and ATP contents, culminating 8-12 h later in cell death. PARP-1 antagonists fully prevented nucleotide depletion and death. Interestingly, in the early 60 min after challenge with N-methyl-N'-nitro-N-nitrosoguanidine, mitochondrial membrane potential and superoxide production significantly increased, whereas cellular ADP contents decreased. Again, these events were prevented by PARP-1 inhibitors, suggesting that PARP-1 hyperactivity leads to mitochondrial state 4 respiration. Mitochondrial membrane potential collapsed at later time points (3 h), when mitochondria released apoptosis-inducing factor and cytochrome c. Using immunocytochemistry and targeted luciferase transfection, we found that, despite an exclusive localization of PARP-1 and poly(ADP-ribose) in the nucleus, ATP levels first decreased in mitochondria and then in the cytoplasm of cells undergoing PARP-1 activation. PARP-1 inhibitors rescued ATP (but not NAD(H) levels) in cells undergoing hyper-poly(ADP-ribosyl)ation. Glycolysis played a central role in the energy recovery, whereas mitochondria consumed ATP in the early recovery phase and produced ATP in the late phase after PARP-1 inhibition, further indicating that nuclear poly(ADP-ribosyl)ation rapidly modulates mitochondrial functioning. Together, our data provide evidence for rapid nucleus mitochondria cross-talk during hyper-poly(ADP-ribosyl)ation-dependent cell death. PMID- 15750181 TI - Independent repression of bile acid synthesis and activation of c-Jun N-terminal kinase (JNK) by activated hepatocyte fibroblast growth factor receptor 4 (FGFR4) and bile acids. AB - The fibroblast growth factor (FGF) receptor complex is a regulator of adult organ homeostasis in addition to its central role in embryonic development and wound healing. FGF receptor 4 (FGFR4) is the sole FGFR receptor kinase that is significantly expressed in mature hepatocytes. Previously, we showed that mice lacking mouse FGFR4 (mR4(-/-)) exhibited elevated fecal bile acids, bile acid pool size, and expression of liver cholesterol 7alpha-hydroxylase (CYP7A1), the rate-limiting enzyme for canonical neutral bile acid synthesis. To prove that hepatocyte FGFR4 was a negative regulator of cholesterol metabolism and bile acid synthesis independent of background, we generated transgenic mice overexpressing a constitutively active human FGFR4 (CahR4) in hepatocytes and crossed them with the FGFR4-deficient mice to generate CahR4/mR4(-/-) mice. In mice expressing active FGFR4 in liver, fecal bile acid excretion was 64%, bile acid pool size was 47%, and Cyp7a1 expression was 10-30% of wild-type mice. The repressed level of Cyp7a1 expression was resistant to induction by a high cholesterol diet relative to wild-type mice. Expression of CahR4 in mR4(-/-) mouse livers depressed bile acid synthesis below wild-type levels from the elevated levels observed in mR4(-/ ). Levels of phosphorylated c-Jun N-terminal kinase (JNK), which is part of a pathway implicated in bile acid-mediated repression of synthesis, was 30% of wild type levels in mR4(-/-) livers, whereas CahR4 livers exhibited an average 2-fold increase. However, cholate still strongly induced phospho-JNK in mR4(-/-) livers. These results confirm that hepatocyte FGFR4 regulates bile acid synthesis by repression of Cyp7a1 expression. Hepatocyte FGFR4 may contribute to the repression of bile acid synthesis through JNK signaling but is not required for activation of JNK signaling by bile acids. PMID- 15750182 TI - Patterning across the ascidian neural plate by lateral Nodal signalling sources. AB - Ascidians are invertebrate chordates with a simple larval tadpole form containing a notochord and an overlying dorsal neural tube. As in vertebrates, the neural tube of ascidian larvae displays positional differences along the rostral-caudal and dorsal-ventral axes in terms of neuronal cell types generated, morphology and gene expression. However, how these differences are established in this simple chordate remains largely unknown. In this study, we show that a single blastomere named b6.5, which is situated in a lateral position in the 32-cell-stage embryo, is a source of signal(s) required for patterning across the medial-lateral axis (future ventral-dorsal axis) of the neural plate. We identify this signal as a Ciona homologue of Nodal, Ci-Nodal. Transcriptional activation of Ci-Nodal in b6.5 depends upon vegetally derived Ci-FGF9/16/20. Using three distinct reagents to inhibit Nodal signals, we show that Nodal signalling is required for neural plate patterning across the medial-lateral axis and that, in the absence of this signal, the caudal-lateral part of the neural plate adopts a medial-like fate. Secondary muscle fate is similarly affected. We conclude that specification of the lateral neural plate is initiated by signalling sources laterally flanking the neural plate and involves a cell-fate choice between lateral and medial neural fates, with Nodal signalling promoting lateral fate. This role for Nodal signalling during ascidian neural plate patterning contrasts with that in vertebrates, where it is implicated in promoting a medial neural fate, the floor plate. PMID- 15750183 TI - Progenitors resume generating neurons after temporary inhibition of neurogenesis by Notch activation in the mammalian cerebral cortex. AB - The mammalian cerebral cortex comprises six layers of neurons. Cortical progenitors in the ventricular zone generate neurons specific to each layer through successive cell divisions. Neurons of layer VI are generated at an early stage, whereas later-born neurons occupy progressively upper layers. The underlying molecular mechanisms of neurogenesis, however, are relatively unknown. In this study, we devised a system where the Notch pathway was activated spatiotemporally in the cortex by in vivo electroporation and Cre-mediated DNA recombination. Electroporation at E13.5 transferred DNA to early progenitors that gave rise to neurons of both low and upper layers. Forced expression of a constitutively active form of Notch (caNotch) at E13.5 inhibited progenitors from generating neurons and kept progenitors as proliferating radial glial cells. After subsequent transfection at E15.5 of a Cre expression vector to remove caNotch, double-transfected cells, in which caNotch was excised, migrated into the cortical plate and differentiated into neurons specific to upper layers. Bromodeoxyuridine-labeling experiments showed that the neurons were born after Cre transfection. These results indicate that cortical progenitors that had been temporarily subjected to Notch activation at an early stage generated neurons at later stages, but that the generation of low-layer neurons was skipped. Moreover, the double-transfected cells gave rise to upper-layer neurons, even after their transplantation into the E13.5 brain, indicating that the developmental state of progenitors is not halted by caNotch activity. PMID- 15750184 TI - The zinc finger transcriptional repressor Blimp1/Prdm1 is dispensable for early axis formation but is required for specification of primordial germ cells in the mouse. AB - Blimp1, a zinc-finger containing DNA-binding transcriptional repressor, functions as a master regulator of B cell terminal differentiation. Considerable evidence suggests that Blimp1 is required for the establishment of anteroposterior axis formation and the formation of head structures during early vertebrate development. In mouse embryos, Blimp1 is strongly expressed in axial mesendoderm, the tissue known to provide anterior patterning signals during gastrulation. Here, we describe for the first time the defects caused by loss of Blimp1 function in the mouse. Blimp1 deficient embryos die at mid-gestation, but surprisingly early axis formation, anterior patterning and neural crest formation proceed normally. Rather, loss of Blimp1 expression disrupts morphogenesis of the caudal branchial arches and leads to a failure to correctly elaborate the labyrinthine layer of the placenta. Blimp1 mutant embryos also show widespread blood leakage and tissue apoptosis, and, strikingly, Blimp1 homozygous mutants entirely lack PGCs. At the time of PGC allocation around 7.25 days post coitum, Blimp1 heterozygous embryos exhibit decreased numbers of PCGs. Thus Blimp1 probably acts to turn off the default pathway that allows epiblast cells to adopt a somatic cell fate, and shifts the transcriptional program so that they become exclusively allocated into the germ cell lineage. PMID- 15750185 TI - Cited2 is required both for heart morphogenesis and establishment of the left right axis in mouse development. AB - Establishment of the left-right axis is a fundamental process of vertebrate embryogenesis. Failure to develop left-right asymmetry leads to incorrect positioning and morphogenesis of numerous internal organs, and is proposed to underlie the etiology of several common cardiac malformations. The transcriptional modulator Cited2 is essential for embryonic development: Cited2 null embryos die during gestation with profound developmental abnormalities, including cardiac malformations, exencephaly and adrenal agenesis. Cited2 is also required for normal establishment of the left-right axis; we demonstrate that abnormal heart looping and right atrial and pulmonary isomerism are consistent features of the left-right-patterning defect. We show by gene expression analysis that Cited2 acts upstream of Nodal, Lefty2 and Pitx2 in the lateral mesoderm, and of Lefty1 in the presumptive floor plate. Although abnormal left-right patterning has a major impact on the cardiac phenotype in Cited2-null embryos, laterality defects are only observed in a proportion of these embryos. We have therefore used a combination of high-resolution imaging and three-dimensional (3D) modeling to systematically document the full spectrum of Cited2-associated cardiac defects. Previous studies have focused on the role of Cited2 in cardiac neural crest cell development, as Cited2 can bind the transcription factor Tfap2, and thus affect the expression of Erbb3 in neural crest cells. However, we have identified Cited2-associated cardiac defects that cannot be explained by laterality or neural crest abnormalities. In particular, muscular ventricular septal defects and reduced cell density in the atrioventricular (AV) endocardial cushions are evident in Cited2-null embryos. As we found that Cited2 expression tightly correlated with these sites, we believe that Cited2 plays a direct role in development of the AV canal and cardiac septa. We therefore propose that, in addition to the previously described reduction of cardiac neural crest cells, two other distinct mechanisms contribute to the spectrum of complex cardiac defects in Cited2-null mice; disruption of normal left-right patterning and direct loss of Cited2 expression in cardiac tissues. PMID- 15750186 TI - Differential regulation of Hedgehog target gene transcription by Costal2 and Suppressor of Fused. AB - The mechanism by which the secreted signaling molecule Hedgehog (Hh) elicits concentration-dependent transcriptional responses from cells is not well understood. In the Drosophila wing imaginal disc, Hh signaling differentially regulates the transcription of target genes decapentaplegic (dpp), patched (ptc) and engrailed (en) in a dose-responsive manner. Two key components of the Hh signal transduction machinery are the kinesin-related protein Costal2 (Cos2) and the nuclear protein trafficking regulator Suppressor of Fused [Su(fu)]. Both proteins regulate the activity of the transcription factor Cubitus interruptus (Ci) in response to the Hh signal. We have analyzed the activities of mutant forms of Cos2 in vivo and found effects on differential target gene transcription. A point mutation in the motor domain of Cos2 results in a dominant negative form of the protein that derepresses dpp but not ptc. Repression of ptc in the presence of the dominant-negative form of Cos2 requires Su(fu), which is phosphorylated in response to Hh in vivo. Overexpression of wild-type or dominant negative cos2 represses en. Our results indicate that differential Hh target gene regulation can be accomplished by differential sensitivity of Cos2 and Su(Fu) to Hh. PMID- 15750187 TI - The roles of two C. elegans HOX co-factor orthologs in cell migration and vulva development. AB - Anteroposterior cell migration and patterning in C. elegans are governed by multiple, interacting signaling pathways and transcription factors. In this study, we have investigated the role of ceh-20, the C. elegans ortholog of the HOX co-factor Extradenticle (Exd/Pbx), and unc-62, the C. elegans ortholog of Homothorax (Hth/Meis/Prep), in two processes that are regulated by Hox gene lin 39: cell migration and vulva formation. As in lin-39 mutants, the anterior migrations of neuroblasts in the Q lineage are truncated in Hox co-factor mutants. Surprisingly, though, our findings suggested that the roles of ceh-20 and unc-62 are different from that of lin-39; specifically, ceh-20 and unc-62 but not lin-39 are required for the transmembrane protein MIG-13 to promote anterior migration. To our knowledge, ceh-20 and unc-62 are the only genes that have been implicated in the mig-13 pathway. We find that ceh-20 and unc-62 are also required for several steps in vulva development. Surprisingly, ceh-20 and unc-62 mutants have phenotypes that are starkly different from those of lin-39 mutants. Thus, in this process, too, ceh-20 and unc-62 are likely to have functions that are independent of lin-39. PMID- 15750188 TI - Nuclear integration of positive Dpp signals, antagonistic Wg inputs and mesodermal competence factors during Drosophila visceral mesoderm induction. AB - Tissue induction during embryonic development relies to a significant degree on the integration of combinatorial regulatory inputs at the enhancer level of target genes. During mesodermal tissue induction in Drosophila, various combinations of inductive signals and mesoderm-intrinsic transcription factors cooperate to induce the progenitors of different types of muscle and heart precursors at precisely defined positions within the mesoderm layer. Dpp signals are required in cooperation with the mesoderm-specific NK homeodomain transcription factor Tinman (Tin) to induce all dorsal mesodermal tissue derivatives, which include dorsal somatic muscles, the dorsal vessel and visceral muscles of the midgut. Wingless (Wg) signals modulate the responses to Dpp/Tin along anteroposterior positions by cooperating with Dpp/Tin during dorsal vessel and somatic muscle induction while antagonizing Dpp/Tin during visceral mesoderm induction. As a result, dorsal muscle and cardiac progenitors form in a pattern that is reciprocal to that of visceral muscle precursors along the anteroposterior axis. Our present study addresses how positive Dpp signals and antagonistic Wg inputs are integrated at the enhancer level of bagpipe (bap), a NK homeobox gene that serves as an early regulator of visceral mesoderm development. We show that an evolutionarily conserved bap enhancer element requires combinatorial binding sites for Tin and Dpp-activated Smad proteins for its activity. Adjacent binding sites for the FoxG transcription factors encoded by the Sloppy paired genes (slp1 and slp2), which are direct targets of the Wg signaling cascade, serve to block the synergistic activity of Tin and activated Smads during bap induction. In addition, we show that binding sites for yet unknown repressors are essential to prevent the induction of the bap enhancer by Dpp in the dorsal ectoderm. Our data illustrate how the same signal combinations can have opposite effects on different targets in the same cells during tissue induction. PMID- 15750189 TI - Randomized trial of high loading dose of clopidogrel for reduction of periprocedural myocardial infarction in patients undergoing coronary intervention: results from the ARMYDA-2 (Antiplatelet therapy for Reduction of MYocardial Damage during Angioplasty) study. AB - BACKGROUND: Aggressive platelet inhibition is crucial to reduce myocardial injury and early cardiac events after coronary intervention. Although observational data have suggested that pretreatment with a high loading dose of clopidogrel may be more effective than a conventional dose, this hypothesis has never been tested in a randomized trial. METHODS AND RESULTS: A total of 255 patients scheduled to undergo percutaneous coronary intervention were randomized to a 600-mg (n=126) or 300-mg (n=129) loading regimen of clopidogrel given 4 to 8 hours before the procedure. Creatine kinase MB, troponin I, and myoglobin levels were measured at baseline and at 8 and 24 hours after intervention. The primary end point was the 30-day occurrence of death, myocardial infarction (MI), or target vessel revascularization. The primary end point occurred in 4% of patients in the high loading dose versus 12% of those in the conventional loading dose group (P=0.041) and was due entirely to periprocedural MI. Peak values of all markers were significantly lower in patients treated with the 600-mg regimen (P< or =0.038). Safety end points were similar in the 2 arms. At multivariable analysis, the high loading regimen was associated with a 50% risk reduction of MI (OR 0.48, 95% CI 0.15 to 0.97, P=0.044). An incremental benefit was observed in patients randomized to the 600-mg dose who were receiving statins, with an 80% risk reduction. CONCLUSIONS: Pretreatment with a 600-mg loading dose of clopidogrel 4 to 8 hours before the procedure is safe and, as compared with the conventional 300-mg dose, significantly reduced periprocedural MI in patients undergoing percutaneous coronary intervention. These results may influence practice patterns with regard to antiplatelet therapy before percutaneous revascularization. PMID- 15750191 TI - Oxidative stress in breast cancer. AB - The present study was undertaken to evaluate the place of oxidative stress on breast cancer. Lipid peroxidation as evidenced by malondialdehyde (MDA) and the status of the antioxidants superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were estimated in tissues of 10 fibroadenoma and 40 breast cancer patients. Lipid peroxidation in breast cancer tissues was enhanced compared to nonmalignant tissues (p < 0.001). Similarly, antioxidants SOD (p < 0.001) and GPx (p = 0.007) in tumor tissues significantly were increased. On the contrary, CAT activity was found significantly decreased (p < 0.001). We found that oxidant/antioxidant status was independent from any prognostic factors concerning breast cancer. The results of our study have shown higher oxygen-free radical production and decreased CAT activity support the oxidative stress hypothesis in breast carcinogenesis. PMID- 15750190 TI - A clinical and biological overview of gastrointestinal stromal tumors. AB - In the last few years a body of knowledge has been generated on the molecular basis of gastrointestinal stromal tumors (GIST). These mesenchymal tumors are characterized by the expression of KIT protein and because they have an activating mutation in a class III receptor tyrosine kinase gene (KIT or PDGFRA). Several KIT-activating mutations, which are largely responsible for the development of this tumor, promote cell survival, proliferation, and migration through different pathways such as MAPK p42/44, AKT, S6K, STAT1, and STAT3. Likewise, gene-activating mutations in the gene PDGFRalpha which codes for the receptor tyrosine kinase, Platelet-derived growth factor receptor alpha have been identified in GIST lacking KIT mutations. This means that KIT and PDGFRalpha mutations appear to be alternative and mutually exclusive oncogenic pathways for GIST development. These tumors may occur anywhere along the gastrointestinal tract (GI). The most frequently involved sites are stomach and small intestine. They are typically chemo- and radioresistant. The discovery of a specific inhibitor of this tyrosine kinase, imatinib mesylate, has radically changed the prognosis of patients with unresectable disease. Only 4 yr after the first patient was successfully treated with imatinib, multiple phase II and III trials have been published and, currently, imatinib mesylate is the only effective systemic treatment available of these tumors. Response rates are approximately 70 90% with acceptable toxicity. GIST are the first model of a solid tumor efficiently treated with a molecular-targeted agent. This review summarizes the clinical and biological aspects of this unique neoplasm. PMID- 15750192 TI - A descriptive study of second primary malignancies associated to breast cancer in a mexican Hispanic population. AB - Breast cancer is the second most frequent tumor in Mexico. Patients diagnosed with this cancer have a higher risk of developing a second malignancy. The objective of our study was to see the frequency, types of second cancers, and its impact on survival, in order to be able to deliver a proper and efficient follow up to these patients, because our patients differ from the population of breast cancer in the rest of the world. Our patients are younger and therefore at higher risk. The clinical records of breast cancer patients treated at the Instituto Nacional de Cancerologia Mexico from 1983 to 1992 were reviewed. In 1370 evaluable patients, 77 (5.6%) developed a second neoplasm, of those, 56 (72.7%) in the contralateral breast and 21 in other sites (27.3%), thyroid was the most frequent followed by ovary and endometrium. Mean age of the patients was 51.5 yr, 45.5 for the other breast and 55.5 for other malignancies (p = 0.01). Median survival for all the group was of 180 mo (3-238). Patients were significantly younger in the contralateral breast group, although all our breast cancer patients are younger. The most frequent second malignancy after the other breast, was thyroid followed by ovary and endometrium with similar survival for both groups. PMID- 15750193 TI - Her-2/neu expression as a predictive factor for response to anthracycline-based chemotherapy in a mexican population of locally advanced breast cancer patients. AB - Breast cancer is the second most frequent tumor in Mexico 30-50% are diagnosed in locally advanced stages requiring neoadjuvant chemotherapy. The overexpression of HER-2/neu has been identified as a chemotherapy response predictor. The objective of our study was to identify response predictive factors to anthracycline-based neoadjuvant chemotherapy in locally advanced breast cancer. Data were collected from clinical records of patients with neoadjuvant anthracycline-based chemotherapy, for clinical stage III breast cancer from 1992 to 1997. Paraffin blocks were reviewed to determine histologic grade, HER-2/neu expression, and ploidy. Patients were divided in two groups: A, 56 cases responding to chemotherapy and, B, 20 nonresponders. Mean age was 50.1 and 45.4 for groups A and B, respectively (OR 7.02, p = 0.004), and those premenopausal were 43% and 70%, respectively (OR 3.1, p = 0.04). Mean tumor size was 5 cm in responders and 8 cm for nonresponders (OR 4, p = 0.02). Clinical stage III-B 16% and 70% for groups A and B (OR 12.2, p = 0.000); tumors were aneuploid in 39% of responders and 18.7% for nonresponders. HER-2 /neu was overexpressed in 64.2% and 50% for groups A and B (OR 3.6, p = 0.06). On multivariate analysis significance was conserved only for age, clinical stage, and size. HER-2/neu lost significance. Age and size were importantly related to tumor response, a higher percentage of HER-2/neu overexpression was observed in responders, without significance due to small sample size. It would be important to study a larger number of patients to obtain more conclusive results. PMID- 15750195 TI - Serum matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 in patients with malignant melanoma. AB - Degradation of basement membranes and extracellular matrix is an essential step in cancer invasion and metastasis. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play key roles in this step. The present study was conducted to investigate the levels of MMP-3 and TIMP-1 in serum of patients with malignant melanoma and the relationship to tumor progression and known prognostic parameters. Seventy patients with cutaneous malignant melanoma were investigated. Serum samples were obtained on first admission before any adjuvant and metastatic treatment was given or follow-up of patients. Serum TIMP-1 and MMP-3 levels were determined by the solid-phase sandwich ELISA (Oncogene Science Inc.) method. The elevation of serum MMP-3 and TIMP-1 levels between the patients with malignant melanoma and healthy controls were not significantly different (p > 0.05). The serum levels of MMP-3 were significantly different in males and females (p = 0.001) and serum TIMP levels were influenced by age (p = 0.047). Except for the ulceration status of the tumor, serum levels of MMP-3 and TIMP-1 were not related to the known prognostic factors such as tumor histology, localization, stage of the disease, Breslow thickness, Clark invasion, mitosis, TIL, and regression of tumor (p > 0.05). In patients with ulceration positive, the serum levels of MMP-3 were higher (p = 0.04) and TIMP-1 were lower (p = 0.008) than those in patients without ulceration. No significant relationship was found between serum levels of MMP-3 and TIMP-1. In conclusion, these results suggest that neither of the serum levels of MMP-3 and TIMP-1 could be a good indicator of invasion and metastasis nor can be recommended as a tumor marker in the management of melanoma patients owing to lack of sensitivity and specificity. However, much research still continues in this field and exciting new knowledge will ultimately emerge. PMID- 15750194 TI - The role of cystatin C and the angiogenic cytokines VEGF and bFGF in patients with esophageal carcinoma. AB - Angiogenesis is the formation of new blood vessels out of the existing vascular bed. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are potent circulating angiogenic factors, whereas cystatin C is one of the most important extracellular inhibitors of several cysteine proteinases. Because proteases degrade interstitial connective tissue and basement membranes during tumor growth and metastasis, an association between cystatin C and the angiogenic factors seems plausible. The primary aim of the present study was to investigate if such a correlation exists between these serum markers. The secondary aim was to determine the prognostic value of these circulating cytokines and cystatin C, collected prior to therapy, in patients with esophageal carcinoma.A total of 42 patients with esophageal carcinoma donated serum samples prior to therapy. VEGF and bFGF were correlated to platelet and leukocyte counts and VEGF was correlated to tumor volume (p = 0.04), whereas bFGF was not (p = 0.08). VEGF was significantly correlated with cystatin C (p = 0.027). Survival analysis showed that VEGF regarded as a continuous variable was associated with a significantly poorer survival in the univariate analysis (p = 0.023); however, this was not found for bFGF (p = 0.46). Neither of the angiogenic factors were associated with survival in the multivariate analysis. In the univariate analysis, cystatin c was correlated with survival (p = 0.01), but this was not found in the multivariate analysis (p = 0.28). In conclusion, VEGF was correlated with cystatin C, possible explanations being discussed in the present article. Results of the present study indicate that use of the angiogenic factors as prognostic factors, prior to therapy in patients with esophageal carcinoma, appears limited. PMID- 15750196 TI - Gamma knife radiosurgery in brain metastases from testicular tumors. AB - To our knowledge, there are no published reports on the effectiveness of radiosurgery in the management of brain metastases from testicular nonseminomatous germ cell tumor. The authors evaluate the results of gamma knife (GK) treatment in three patients with these unusual intracranial lesions. Between April 1995 and July 2001, three patients with brain metastasis from testicular nonseminomatous germ cell tumor underwent adjuvant radiosurgery at our department. The primary tumor had been surgically removed in all cases. At diagnosis, one patient was stage IB and two were stage III poor risk. Chemotherapy and whole brain radiotherapy were administered before radiosurgery in all cases. Pre-GK radiotherapy was administered with a daily fraction dosage of 1.8-2.0 Gy. The indications for radiosurgery were tumor volume <20 cm3, microsurgery too risky, refusal of surgery. All the lesions were located in eloquent brain areas. Post-GK high-dose chemotherapy with autologous peripheral blood stem-cell rescue was administered in two cases due to systemic recurrence of the disease. All patients are still alive with a median and mean follow-up period after radiosurgery of 63 and 68.3 mo, respectively. They had no neurological deficits at the latest examination. Neuroradiological follow-up invariably showed tumor growth control (complete response in two cases and partial response in one) with typically delayed post-radiosurgical imaging changes (transient in two cases and long-lasting in one). In conclusion, GK seems to be highly effective and safe in brain metastases from testicular nonseminomatous germ cell tumor. In cases with diffuse metastatic brain involvement, the whole brain radiotherapy preceding radiosurgery should be delivered with 1.8 Gy daily fraction to prevent the risk of long-lasting post radiosurgical imaging changes. PMID- 15750197 TI - Mucosa-associated lymphoid tissue (MALT) lymphoma of the stomach: results of a controlled clinical trial. AB - Treatment of patients with early stage gastric mucosa-associated lymphoid tissue (MALT) remains undefined. We began a controlled clinical trial to evaluate efficacy and toxicity of the most common therapies. Two hundred and forty-one patients with gastric low-grade MALT lymphoma in early stage (IE and IIE) were randomized to surgery (80 cases), radiotherapy (78 cases), and chemotherapy (83 cases). With a median follow-up of 7.5 yr, actuarial curves at 10 yr showed that event-free survival was 52% in patients treated with surgery, 52% in radiotherapy arm, and 87% in the chemotherapy group (p < 0.01). However, overall survival did not showed any statistical differences: 80%, 75% and 87%, respectively (p = 0.4). Acute and late toxicities were mild. No death-related treatments were observed. No clear differences were observed between the most common therapies in patients with primary gastric MALT lymphoma in early stages, probably because this type of lymphoma has an high response rate to salvage treatment after failure to local treatment (surgery and radiotherapy). Thus considered, chemotherapy alone is an effective and safe therapeutic approach in this setting of patients. Surgery or radiotherapy will be reserved to patients that are not candidates for chemotherapy. PMID- 15750198 TI - Expression of cyclooxygenase-2 and inducible nitric oxide synthase correlates with tumor angiogenesis in endometrial carcinoma. AB - The present study evaluated the significance of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in 30 patients with endometrial carcinoma and the relationship of those molecular markers to tumor characteristics and microvessel density (MVD). Immunohistochemical expression of COX-2, iNOS, and CD34 was analyzed on paraffin-embedded tissue sections. The COX-2 and iNOS positive rates were 66.7% and 73.3%, respectively. The level of COX-2 expression was higher in grade II tumors than in grade III tumors (p < 0.05). The percentage of iNOS positivity was higher in patients with deep myometrial invasion than in patients without or less than 50% myometrial invasion (p < 0.05). There was significant correlation between positive COX-2 and positive iNOS expression (r = 0.601, p < 0.001). Both COX-2 and iNOS were significantly correlated with MVD (r = 0.02 p < 0.05; r = 0.599 p < 0.0001, respectively). The present findings suggest that combined expression of COX-2 and iNOS may play an important role in development and invasion of endometrial cancer and that this could be partially attributable to modulation of angiogenesis by COX-2 and iNOS. PMID- 15750199 TI - Bone marrow hyaluronan distribution in patients with acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is a clonal disorder characterized by abnormal proliferation of myeloid cell precursors. Research has mainly focused on the cellular events, but the bone marrow matrix has attracted minor interest. In this study bone marrow biopsies were obtained from 35 newly diagnosed AML patients. The bone marrows were analyzed regarding the occurrence and distribution of hyaluronan (HYA) and reticulin fibers (type III collagen). The bone marrow sections were analyzed histochemically and compared with bone marrows from 30 healthy controls. The HYA staining was significantly stronger in the AML patients compared with the controls. Only one patient demonstrated abnormal reticulin staining score, but in the group of patients with antecedent myelodysplastic syndrome (MDS), the reticulin staining score was significantly higher compared with the patients with de novo AML. There was a significant correlation between the HYA staining and reticulin staining scores in the AML patients as was seen in the control group. PMID- 15750200 TI - Methods of dendritic cell preparation for acute lymphoblastic leukemia immunotherapy in children. AB - Cell immunotherapy through dendritic cells (DC) presents a hopeful strategy for the treatment of various tumors. The aim of our study was to find which progenitor cells are most suitable for the preparation of dendritic cells in acute lymphoblastic leukemia (ALL) in pediatric patients, whether blasts from bone marrow or dendritic cells generated from peripheral blood mononuclear cells taken at the time of remission after induction chemotherapy. DC generated from the BM blasts of patients with B-ALL and T-ALL (n = 15) at the time of diagnosis expressed low levels of costimulatory molecules and CD markers typical for mature DC. In contrast, DC cultivated from peripheral mononuclear cells of patients (n = 9) had comparable morphology and expression of costimulatory molecules to DC obtained from healthy individuals, which was even higher after tumor lysate pulsing. Autologous lymphocyte proliferation increased after DC blasts lysate pulsation and further after lymphocyte restimulation, showing evidence of induction of specific cytotoxic lymphocytes. When comparing both cell sources for the preparation of DC in patients with ALL, it appears that peripheral mononuclear cells obtained after chemotherapy are more suitable than bone marrow leukemic blasts due to similar morphology, phenotypic, and functional capacity to monocytes of healthy donors. Despite this, it is necessary to take into account individual variability when preparing DC-based vaccines. The final verification of the efficiency of immunotherapy against residual hematopoietic malignant cells in patients with ALL can only be obtained through a clinical study. PMID- 15750203 TI - The genetic basis of sporadic pancreatic cancer. PMID- 15750201 TI - Ten years of disease-free survival between two diagnoses of small-cell lung cancer: a case report and a literature review. AB - Small-cell lung carcinoma (SCLC) represents one-fifth of all cases of bronchopulmonary cancer and has a 5-yr survival of 2-4%. Long-term survivors of SCLC are at risk for developing second primary aerodigestive tumors. We report a case of a long-term survivor who had a 10-yr disease-free survival between two diagnoses of SCLC. A literature review identified four case reports and seven review series with a total of 26 cases of 5-yr disease-free survivors of SCLC who developed a second SCLC. A total of 4574 patients were reported in the review series. Five-year disease-free survival was documented in 139 of 4574 patients. Twenty-two (15.8%) of those developed a second SCLC over the next 7 yr of follow up, with an averaged annual incidence equal to fivefold that of the general population. Earlier reports of a slow radiographic doubling time for some cases of SCLC suggests that survivors of SCLC may still develop a recurrent SCLC following 12 yr of disease-free follow up. It remains difficult to ascertain whether a second SCLC is a recurrence or a second primary tumor in the absence of a preneoplastic lesion for SCLC. New genetic markers may hold the answer. They may also help screen high-risk patients including survivors of SCLC. PMID- 15750204 TI - Report on the Dutch consensus development meeting for implementation and further development of population screening for colorectal cancer based on FOBT. AB - A consensus development meeting was held to evaluate whether or not in the Netherlands all requirements were fulfilled for implementation of population screening with FOBT for colorectal cancer, or whether consensus was present that fulfilment by additional research or organisational actions could be obtained within 2-3 years. There was consensus that all classical Wilson and Jungner (1968) criteria, and six additional ones added more recently, had already been fulfilled or could be fulfilled within 2-3 years. Consequently, it was concluded that a national population screening for colorectal cancer should be implemented and carried out in the Netherlands in line with current national and European cancer screening programmes. A list of organisational actions to be taken was established. Research that is needed before the actual national launch of the screening within 2-3 years has been defined. Priorities have to be set for research and organisational actions for the coming 2-3 years for the implementation of population screening. In addition, research suggestions have been defined for the next 10-15 years for evaluation and/or improvement of implemented FOBT screening, and for future screening methodology. It was considered essential that infrastructure for future research would be embedded in the screening programme. A project group to arrange this should be formed. PMID- 15750205 TI - Detection of cervical cancer and high grade neoplastic lesions by a combination of liquid-based sampling preparation and DNA measurements using automated image cytometry. AB - OBJECTIVE: To establish if measurements of DNA ploidy could be used to assist cytopathologists and cytotechnologists in population based cervical cancer screening programs in countries where manually reading the slides is impossible due to the lack of sufficient skilled cytotechnologists. The goal of such program is to identify only clinically significant lesions, i.e. those where a clinical intervention to remove the lesion is required immediately. STUDY DESIGN: A total of 9905 women were enrolled in the study. Cervical samples were taken with a cervix brush that was then placed into a fixative solution. The cells were separated from mucus by mechanical and chemical treatment and then deposited onto microscope slides by a cytocentrifuge. Two slides were prepared from each case; one slide was stained by Papanicolaou stain for manual cytology examination, while the other slide was stained by a DNA specific stain. The latter slide was used to determine the relative amount of DNA in the cell nuclei. RESULTS: A total of 876 women were followed by colposcopy examination where biopsies were taken from the visible lesions or from suspicious areas and histopathology diagnosed 459 as normal or benign cases, 325 as CIN1, 36 as CIN2, 25 as CIN3/CIS, and 31 as invasive cancer. Of these 876 cases, manual cytology called 655 normal or ASCUS, 197 as LSIL, 16 cases as HSIL, and 8 as cancer. DNA measurements found 704 cases having no cells with DNA greater than 5c, 98 cases where there were 1 or 2 cells having DNA amount greater than 5c, and 74 cases where there were 3 or more cells having DNA amount greater than 5c. If manual cytology were to be used to refer all cases of HSIL and cancer to colposcopy and biopsy, 23 lesions that had to be removed would have been discovered (2 CIN2, 11 CIN3/CIS, and 10 cancers), for a sensitivity of 25.0+/-5.2% at specificity of 99.9+/-0.1%. If DNA assisted cytology were to be used instead, and all cases having 3 or more cells with DNA amount greater than 5c were to be referred to colposcopy and biopsy, then 50 lesions that had to be removed would have been discovered (10 CIN2, 15 CIN3/CIS and 25 cancers) for the sensitivity of 54.3+/-6.2% at specificity of 96.9+/-0.6%. CONCLUSIONS: The study suggests that screening for high grade cervical neoplastic lesions and cervical cancer by DNA assisted cytology could be implemented with minimal use of skilled cytotechnologists, at least in those countries where it would be difficult to introduce population based screening for cervical cancer due to the lack of availability of such skilled cytotechnologists. PMID- 15750206 TI - Hypoxia-induced acidification causes mitoxantrone resistance not mediated by drug transporters in human breast cancer cells. AB - Hypoxia has clinically been associated with resistance to chemotherapy. The aim of this study was to investigate whether hypoxia induces resistance to doxorubicin and mitoxantrone, two common drugs in cancer treatment, in MCF-7 breast cancer cells, and SW1573 non-small lung cancer cells. In addition, the role of drug transporters P-gp, BCRP and MRP1 was analysed. Hypoxia induced resistance in MCF-7 cells to mitoxantrone shifted the IC(50) value from 0.09 microM (+/-0.01) to 0.54 microM (+/-0.06) under hypoxia, whereas survival of MCF 7 and SW1573 cells in the presence of doxorubicin was not altered. Accumulation of mitoxantrone and daunorubicin, a doxorubicin fluorescent homologue, appeared to be 5.3 and 3.2 times lower in MCF-7 cells, respectively. Cytotoxicity assays showed no increased functionality of the drug transporters P-gp, BCRP and MRP1 under hypoxia. In addition, protein levels of these drug transporters were not changed. Medium of the MCF-7 cells became more acidic under hypoxia thereby causing a decreased uptake of mitoxantrone. Hypoxia induces mitoxantrone resistance in MCF-7 cells not mediated by the three major MDR transporters. Hypoxia-induced acidification may cause this resistance by decreased cellular uptake together with a lowered cytotoxicity due to pH-dependent topoisomerase type II activity. PMID- 15750207 TI - Different DNA ploidy patterns for the differentiation of common subtypes of renal tumors. AB - OBJECTIVES: The common subtypes of renal tumors are conventional or clear cell carcinoma, papillary carcinoma, chromophobe carcinoma and oncocytoma. Each subtype has its distinct histogenesis and clinical evolution. DNA ploidy is viewed as a marker of gross genomic aberrations. The aim of this study is to evaluate the DNA ploidy in the common subtypes of renal tumors to increase our understanding of renal tumor biology and to broaden clinical application of DNA ploidy. METHODS: 38 renal tumor samples (13 clear cell RCCs, 12 papillary RCCs, 7 chromophobe RCCs, and 6 oncocytomas) were studied. Five biopsies of different parts of each fresh tumor were subjected to a flow cytometric analysis of DNA ploidy. RESULTS: All tumors except one papillary RCC generated interpretable DNA histograms. Flow cytometric analysis of oncocytomas showed the diploid pattern (29/30 frequencies) while the chromophobe RCC never showed the diploid pattern (0/55 frequencies) (p<0.01). 3/7 chromopbobe RCCs possessed the hypodiploid stemline. The hypodiploid stemline appeared neither in conventional RCCs (0/63 frequencies) nor in papillary RCCs (0/50 frequencies). The diploid pattern was dominant in conventional and papillary RCCs. 10/13 (76.9%) of clear cell RCCs and 9/11 (81.8%) of papillary RCCs possessed a homogeneous DNA ploidy pattern while only 1/7 (14.3%) has a homogeneous DNA ploidy pattern. 6/7 chromophobe RCCs had multiple aneuploid stemlines. CONCLUSIONS: Flow cytometric analysis reveals that conventional and papillary RCCs are more homogeneous than chromophobe RCC. Each subtype of renal tumors possesses a specific DNA ploidy pattern. The analysis of DNA ploidy is useful for the differentiation of common subtypes of renal tumors in morphologically difficult cases. PMID- 15750208 TI - DNA copy number changes at 8q11-24 in metastasized colorectal cancer. AB - BACKGROUND: C-Myc, a well-known oncogene located on 8q24.12-q24.23, is often amplified and over-expressed in both primary and metastasizing colorectal cancer. In addition, PRL-3 (also known as PTP4A3), a tyrosine phosphatase located on 8q24.3, is amplified in colorectal cancer metastasis. Beside PRL-3 and c-myc, other oncogenes located on the 8q23-24 region might be involved in this process. Therefore, the present study aims to correlate DNA copy number status of a series of genes at 8q23-24 in colorectal cancer at high resolution in correlation to metastatic disease. MATERIALS AND METHODS: Thirty-two cases of colorectal cancer, 10 stage B1, 10 B2 and 12 D (Astler-Coller) with their corresponding liver metastasis and one colorectal cell line (colo205, previously analyzed by array CGH), were included in this study. A chromosome 8 specific MLPA probe mixture was used to analyze the presence of DNA copy number changes. The probe mixture contained 29 probes covering 25 genes on chromosome 8, as well as 6 control probes on other chromosomes. RESULTS AND DISCUSSION: MLPA results obtained of the colo205 colorectal cell line were comparable with previous array-CGH results, thus validating the MLPA probe mixture. Astler-Coller B1 and B2 colorectal cancers differed significantly in DNA copy number of the genes, MOS (p=0.04), MYC (p=0.007), DDEF1 (p=0.004), PTK2 (p=0.02) and PTP4A3 (p=0.04). When comparing these with Astler-Coller D primary tumors, significant differences were seen for several genes as well (MYC (p<0.000), DDEF1 (p<0.000), SLA (p<0.000), PTK2 (p<0.000), PTP4A3 (p=0.002), and RECQL4 (p=0.01)). When comparing primary Astler Coller D tumors and their corresponding liver metastases, a similar pattern of gains and losses was observed. Most of the liver metastases showed higher DNA copy number ratios than the corresponding primary tumors, but this difference was only significant for TPD52 (p=0.02) and EIF3S6 (p=0.007). CONCLUSION: In addition to c-myc, multiple genes on chromosome 8 differed significantly between primary colorectal cancers with and without liver metastases. This observation is consistent with the concept that clinical behaviour, like risk of liver metastasis, is determined by the genomic profile that is already present in the primary tumor. PMID- 15750209 TI - Feedback on a feedback loop: the hypothalamic-pituitary-gonadal axis. PMID- 15750210 TI - Apoptotic effect of caspase-3 cleaved tau in hippocampal neurons and its potentiation by tau FTDP-mutation N279K. AB - Pathological changes in the microtubule associated protein tau are a major hallmark of many human dementias collectively defined as tauopathies. In familiar frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), several mutations in the tau gene have been identified showing that primary malfunction of tau can lead to neurodegeneration. In addition to mutation at genetic level, a number of post-translational modifications of tau occur in tauopathies, including abnormal phosphorylation and aberrant proteolysis described in Alzheimer's Disease (AD). The presence of cleaved tau in AD neurons is associated with expression of markers for neuronal death. According to our previous work, tau is a substrate for the apoptotic protease caspase-3 that turns tau itself into an effector of apoptosis (tau cleaved at D-421), generating a positive-feedback loop that is self-propagating. Cleavage of tau by caspase-3 was recently confirmed to occur in AD brain as an early event. Here we show the apoptotic properties of tau fragment tau151-421 in primary cultures of rat hippocampal neurons; such cellular model is of special interest considering the selective vulnerability of hippocampal neurones in AD. The apoptotic capacity of tau151-421 is markedly enhanced by both treatment with amyloid peptide Abeta25 35, and the FTDP-17 tau mutation N279K. PMID- 15750211 TI - Proteasome or calpain inhibition does not alter cellular tau levels in neuroblastoma cells or primary neurons. AB - Impaired tau catabolism may contribute to tau accumulation and aggregation in Alzheimer's disease and neurofibrillary tangle formation. This study examined the effects of proteasome and calpain inhibition on tau levels and turnover in primary rat hippocampal neurons and differentiated SH-SY5Y human neuroblastoma cells. Administration of proteasome (MG-115, lactacystin) or calpain (MDL28170) inhibitors for up to 24 hours did not alter tau levels in differentiated SH-SY5Y cells or rat hippocampal neurons. Addition of 1 microM and 10 microM MG-115 did not change total tau levels, but did result in increased reactivity of phosphorylation-dependent tau antibodies (PHF-1, CP-13) and decreased Tau-1 immunoreactivity. Administration of cycloheximide to inhibit de novo protein synthesis also did not alter tau levels in the presence or absence of lactacystin. These results demonstrate that although the proteasome and calpain protease systems are capable of degrading tau in cell-free assays, their inhibition does not alter cellular tau levels in primary neurons or differentiated neuroblastoma cells. PMID- 15750212 TI - Isoform-specific effects of apolipoprotein E on secretion of inflammatory mediators in adult rat microglia. AB - Inflammation mediated by activated microglia cells has been shown to contribute to the pathogenesis of Alzheimer disease (AD) [1]. Microglia are the immune cells in the central nervous system, and when activated they secrete the lipid-derived mediator prostaglandin E2 (PGE2), the cytokine interleukin-1beta (IL-1beta), and other inflammatory mediators. Apolipoprotein E isoform 4 (apoE4), coded for by the gene APOE4 (epsilon4), has been shown to correlate with higher risk of onset of AD, as well as with increased severity of other diseases with a neuroinflammatory component. This study investigated isoform-specific effects of apoE on the regulation of PGE2, COX2, and IL-1beta expression. Two physiologically relevant preparations of apoE displayed an isoform-specific effect on inflammation in primary adult microglia cultured from adult rat brain cortex. Specifically, apoE4 alone, but not the more common isoform apoE3, stimulated secretion of PGE2 and IL-1beta. The increase in PGE2 release stimulated by apoE4 was not accompanied by the upregulation of the COX-2 enzyme in microglia. PMID- 15750213 TI - Fibrillar amyloid beta-protein inhibits the activity of high molecular weight brain protease and trypsin. AB - The effect of soluble amyloid beta-protein (sAbeta) and fibrillar amyloid beta protein (fAbeta) on the casein-digesting activity of high molecular weight bovine brain protease (HMW protease) and trypsin was studied. While sAbeta stimulated the casein-digesting activity of HMW protease in a concentration-dependent manner, it did not affect trypsin activity. Structure-activity relationship was studied by testing different soluble and fibrillar Abeta peptides. Various Abeta peptides affected casein-digesting activity of HMW protease differently: sAbeta 1 40 > sAbeta 22-35 = sAbeta 1-11 = sAbeta1-16 > sAbeta 1-28 = sAbeta 31-35, while sAbeta 12-28 and sAbeta 25-35 had no effect. On the other hand, among the fibrillar beta peptides, only fAbeta 1-40 significantly inhibited the casein digesting activity of HMW protease. Tricine gel electrophoresis showed that sAbeta was digested by trypsin while it remained un-cleaved in the presence of HMW protease. However, fAbeta, a major component of amyloid plaques in Alzheimer's disease, inhibited the casein-digesting activity of both HMW protease and trypsin. fAbetawas found to be resistant to proteolysis by HMW protease and trypsin. The trypsin resistance starts in the early stage of fibrillization of Abeta, i.e., aggregated Abeta. Taken together, these results suggest that fibrillization of Abeta may affect the clearance of Abeta by inhibiting the brain proteases, thereby increasing the concentration of circulating Abeta, that may further increase the Abeta fibrillization. PMID- 15750214 TI - Review of insulin and insulin-like growth factor expression, signaling, and malfunction in the central nervous system: relevance to Alzheimer's disease. AB - Interest in characterizing the role of impaired insulin actions in Alzheimer's disease (AD) and vascular dementia is growing exponentially. This review details what is currently known about insulin, insulin-like growth factor type I (IGF-I) and IGF-II proteins and their corresponding receptors in the brain, and delineates the major controversies pertaining to alterations in the expression and function of these molecules in AD. The various experimental animal models generated by over-expression, mutation, or depletion of genes that are critical to the insulin or IGF signaling cascades are summarized, noting the degrees to which they reproduce the histopathological, biochemical, molecular, or behavioral abnormalities associated with AD. Although no single model was determined to be truly representative of AD, depletion of the neuronal insulin receptor and intracerebroventricular injection of Streptozotocin reproduce a number of important aspects of AD-type neurodegeneration, and therefore provide supportive evidence that AD may be caused in part by neuronal insulin resistance, i.e. brain diabetes. The extant literature did not resolve whether the CNS insulin resistance in AD represents a local disease process, or complication/extension of peripheral insulin resistance, i.e. chronic hyperglycemia, hyperinsulinemia, and Type 2 diabetes mellitus. The available epidemiological data are largely inconclusive with regard to the contribution of Type 2 diabetes mellitus to cognitive impairment and AD-type neurodegeneration. A major conclusion drawn from this review is that there is a genuine need for thorough and comprehensive study of the neuropathological changes associated with diabetes mellitus, in the presence or absence of superimposed AD or vascular dementia. Strategies for intervention may depend entirely upon whether the CNS disease processes are mediated by peripheral, central, or both types of insulin resistance. PMID- 15750216 TI - Insulin signaling, diabetes mellitus and risk of Alzheimer disease. PMID- 15750215 TI - Impaired insulin and insulin-like growth factor expression and signaling mechanisms in Alzheimer's disease--is this type 3 diabetes? AB - The neurodegeneration that occurs in sporadic Alzheimer's disease (AD) is consistently associated with a number of characteristic histopathological, molecular, and biochemical abnormalities, including cell loss, abundant neurofibrillary tangles and dystrophic neurites, amyloid-beta deposits, increased activation of pro-death genes and signaling pathways, impaired energy metabolism/mitochondrial function, and evidence of chronic oxidative stress. The general inability to convincingly link these phenomena has resulted in the emergence and propagation of various heavily debated theories that focus on the role of one particular element in the pathogenesis of all other abnormalities. However, the accumulating evidence that reduced glucose utilization and deficient energy metabolism occur early in the course of disease, suggests a role for impaired insulin signaling in the pathogenesis of AD. The present work demonstrates extensive abnormalities in insulin and insulin-like growth factor type I and II (IGF-I and IGF-II) signaling mechanisms in brains with AD, and shows that while each of the corresponding growth factors is normally made in central nervous system (CNS) neurons, the expression levels are markedly reduced in AD. These abnormalities were associated with reduced levels of insulin receptor substrate (IRS) mRNA, tau mRNA, IRS-associated phosphotidylinositol 3 kinase, and phospho-Akt (activated), and increased glycogen synthase kinase-3beta activity and amyloid precursor protein mRNA expression. The strikingly reduced CNS expression of genes encoding insulin, IGF-I, and IGF-II, as well as the insulin and IGF-I receptors, suggests that AD may represent a neuro-endocrine disorder that resembles, yet is distinct from diabetes mellitus. Therefore, we propose the term, "Type 3 Diabetes" to reflect this newly identified pathogenic mechanism of neurodegeneration. PMID- 15750217 TI - A new thiophosphate, Rb(0.38)Ag(0.5)Nb2PS10. AB - The structure of the new pentanary thiophosphate rubidium silver diniobium tris(disulfide) tetrathiophosphate, Rb(0.38)Ag(0.5)Nb2PS10, is made up of one dimensional (infinity)1[Nb2PS10-] chains along the [001] direction. These chains are separated from one another by Ag+ and disordered Rb+ ions. The Nb2PS10- chain is built up from bicapped trigonal prismatic Nb(2)S(12) units which lie about inversion centres and tetrahedral PS4 groups. The Nb2S12 units are linked together to form linear Nb2S9 chains by sharing S-S prism edges. Short [2.898 (1) and 2.908 (1) A] and long [3.724 (1) A] Nb...Nb distances alternate along the chains, and S(2)2- and S2- anionic species co-exist in the structure. The Ag(+) cation lies on an inversion centre and has distorted octahedral coordination described as a [2+4]-bonding interaction. PMID- 15750218 TI - Magnesium aluminium chromite. AB - The cation distribution in a natural magnesium aluminium chromite spinel (cubic, space group Fd-3m), Al(0.41)Cr(1.42)Fe(0.65)Mg(0.4)O(4), was determined by electron-microprobe analysis, Mossbauer spectroscopy and single-crystal X-ray analysis. Several structural models of the octahedral and tetrahedral cation distributions were tested; the most probable is (Mg(0.40(11))(2+),Al(0.28(5))(3+),Fe(0.39(4))(2+))[Al(0.13(5))(3+),Cr(1.42(6))(3+ ,Fe(0.26(4))(3+),Phi(0.19)]O(4)(2-), where (...) and [...] represent the tetrahedral and octahedral sites, respectively, and Phi represents a vacancy. PMID- 15750219 TI - High-pressure syntheses of alpha-RE2B4O9 (RE = Sm, Ho), with a structure type displaying edge-sharing BO4 tetrahedra. AB - The compounds alpha-RE2B4O9, with RE = Sm (disamarium tetraborate) and Ho (diholmium tetraborate), were synthesized under conditions of high pressure and high temperature in a Walker-type multianvil apparatus, at 7.5 GPa and 1323 K for alpha-Sm2B4O9 and at 10 GPa and 1323 K for alpha-Ho2B4O9. The crystal structures were determined from single-crystal X-ray diffraction data collected at room temperature. The structures are isotypic with the already known alpha-RE2B4O9 (RE = Eu-Dy) phases, displaying the new structural motif of edge-sharing BO4 tetrahedra next to the known motif of corner-sharing BO4 tetrahedra. As the end members of this isotypic series, the two title compounds mark the borders of the stability field of the appearance of edge-sharing BO4 tetrahedra. PMID- 15750220 TI - Synthesis and ab initio X-ray powder diffraction structure of the new alkali and alkali earth metal borate NaCa(BO3). AB - A sodium calcium borate, NaCaBO3, has been synthesized by the solid-state reaction method and the structure solved from X-ray powder diffraction data. The compound crystallizes in space group Pmmn and has a desired structure type containing isolated planar BO3(3-) anions. Mixed occupancy is found to exist in the Ca site, with partial replacement by Na. One Ca/Na mixed atom and one Na atom are at sites with mm2 symmetry, and a second Ca/Na mixed atom, an Na atom, two B and two O atoms are on mirror planes. PMID- 15750221 TI - Al6Ti2O13, a new phase in the Al2O3-TiO2 system. AB - The compound Al6Ti2O13 (hexaaluminium dititanium tridecaoxide) has been synthesized using an arc-imaging furnace, which allows fast cooling of melted oxides. The structure consists of infinite double chains of polyhedra running along the c axis. These chains are built up by four kinds of strongly distorted oxygen octahedra randomly occupied by either Ti or Al (point symmetry m or m2m), and by trigonal bipyramids exclusively occupied by Al (point symmetry m2m). PMID- 15750222 TI - The two-dimensional coordination polymer poly[[bis(3-methylpyridine)cadmium(II)] tetra-mu-cyano-nickel(II)]. AB - The title complex, [CdNi(CN)4(C6H7N)2]n, adopts a slightly distorted octahedral geometry around the Cd centre. Four cyanide N atoms occupy the equatorial coordination sites around the Cd centre. The structure consists of corrugated and cyanide-bridged polymeric networks made up of tetracyanonickelate ions coordinated to cadmium, with the Ni ion coordinated by four cyanide ligands in a square-planar arrangement. The Cd and Ni atoms occupy special positions of 2/m site symmetry. The 3-methylpyridine group, except for two methyl H atoms, lies on a crystallographic mirror plane. The 3-methylpyridine molecules, bound to cadmium in trans positions, are located on both sides of the network. The bonding in the networks occurs because of a departure of the Ni-C-N-Cd sequence of atoms from linearity at the C and N atoms. PMID- 15750223 TI - trans-Diaquabis(6-hydroxypicolinato-kappa2N,O2)copper(II). AB - In the title compound, [Cu(C6H4NO3)2(H2O)2], the Cu(II) ion lies on an inversion centre and has an elongated octahedral environment, equatorially trans coordinated by two N,O-bidentate picolinate ligands and axially coordinated by two water O atoms. The complex molecules form layers, which are linked by O-H...O hydrogen bonds between the aqua ligands and neighbouring carboxylate groups. An intramolecular hydrogen bond between the coordinated carboxylate O atom and the hydroxy H atom is also observed. PMID- 15750224 TI - Cyano-bridged extended heteronuclear supramolecular architectures with hexacyanocobalt(III) as building blocks. AB - The cyano-bridged heteronuclear coordination polymer poly[tris[(5,12-dimethyl 7,14-diphenyl-1,4,8,11-tetraazacyclotetradeca-4,11-diene)copper(II)]-hexa-mu cyano-bis[tricyanocobalt(III)] dimethylformamide solvate trihydrate], {[Cu3Co2(CN)12(C24H32N4)3].C3H7NO.3H2O}n, was synthesized by the assembly reaction of [CuL]2+ (L is 5,12-dimethyl-7,14-diphenyl-1,4,8,11 tetraazacyclotetradeca-4,11-diene) and [Co(CN)6]3- in a dimethylformamide-water solution. The structure consists of neutral cyano-bridged Cu3Co2 units with the unique Co atom in a general position and all three Cu atoms on independent inversion centres. Each [Co(CN)6]3- ion connects three CuII ions via three cyano groups to form a novel cyano-bridged two-dimensional stair-shaped-layer structure. The water and dimethylformamide molecules are situated in the inter fragment spaces. PMID- 15750225 TI - [(1R,2R)-2-Amino-1,2-diphenyl-N-(p-tolylsulfonyl)ethylamido]chloro(eta(6) ethoxybenzene)ruthenium(II) methanol solvate. AB - The title compound, [Ru(C21H21N2O2S)Cl(C8H10O)].CH4O or [Ru(TsDPEN)Cl(eta6 C6H5OCH2CH3)].CH4O [where TsDPEN is (1R,2R)-1,2-diphenyl-N-(p toluenesulfonyl)ethylenediamine], contains an S-chiral Ru centre in a distorted octahedral environment, with similar bond lengths and angles to analogous complexes. The very short (N-)H...Cl distance of 2.61 A is ascribed to an intramolecular hydrogen bond. PMID- 15750226 TI - catena-Poly[bis[(eta2-1-allyl-3-aminopyridinium)copper(I)]-di-mu-chloro-copper(I) di-mu-chloro-copper(I)-di-mu-chloro]. AB - Crystals of the title pi-complex, [Cu4Cl6(C8H11N2)2]n, were obtained by means of alternating-current electrochemical synthesis. The structure consists of infinite copper-chlorine chains to which 1-allyl-3-aminopyridinium moieties are attached via a eta2 Cu-(C=C) interaction. The two independent Cu atoms have distinct coordination environments. One is three-coordinate, surrounded by two chloro ligands and the olefinic bond, whereas the second copper center is surrounded by a tetrahedral arrangement of four Cl atoms. The lower basicity of 3-aminopyridine as compared with 2- and 4-aminopyridine lowers the capacity of the organic ligand for donating to N-H...Cl hydrogen bonds and results in the formation of a large inorganic fragment. PMID- 15750227 TI - catena-Poly[[bis[aqua(1,10-phenanthroline)lead(II)]-di-mu3-5-carboxy-3 sulfonatobenzoato] dihydrate]. AB - In the centrosymmetric title polymer, catena-poly[[bis[aqua(1,10-phenanthroline kappa2N,N')lead(II)]-di-mu3-5-carboxy-3-sulfonatobenzoato 1:2:1'kappa4O3:O1,O1':O1;2':1:2kappa4O1:O1,O1':O3] dihydrate], {[Pb(C8H4O7S)(C12H8N2)(H2O)].H2O}n, each seven-coordinate lead(II) ion is bound by five O atoms from one water molecule and three 5-sulfoisophthalate (sip) anions, and by two N atoms from a 1,10-phenanthroline (phen) ligand. The sip sulfonate group is monodentate. One O atom of the sip carboxylate group is chelated to one Pb2+ cation, with the other also bridging an adjacent Pb2+ cation. The carboxyl group is uncoordinated. This unusual coordination results in a chain structure along the b axis, which is linked by strong intermolecular hydrogen bonds into a three-dimensional network. PMID- 15750228 TI - Poly[[bis(pyridin-3-ol)manganese(II)]-di-mu-pyridin-3-olato]. AB - In the title complex, [Mn(C5H4NO)2(C5H5NO)2]n or [Mn(mu-3-PyO)2(3-PyOH)2]n (3-PyO is the pyridin-3-olate anion and 3-PyOH is pyridin-3-ol), the Mn(II) atom lies on an inversion centre and has octahedral geometry, defined by two N atoms and two deprotonated exocyclic O atoms of symmetry-related pyridin-3-olate ligands [Mn-N = 2.3559 (14) A and Mn-O = 2.1703 (11) A], as well as two N atoms of terminal 3-PyOH ligands [Mn-N = 2.3482 (13) A]. The Mn(II) atoms are bridged by the deprotonated pyridin-3-olate anion into a layer structure, generating sheets in the (-101) plane. These sheets are linked by O-H...O hydrogen bonds. There are also pi-pi and C-H...pi interactions in the crystal structure. PMID- 15750229 TI - Poly[tetrasodium(I)-tetra-mu2-bis(butane-1,4-diyldioxy)borato-mu2-1,4 butanediol]. AB - In the title compound, [Na4(C8H16BO4)4(C4H10O2)]n, there are two coordination types for the four independent Na+ cations: two Na+ cations bond to six diolate O atoms [Na-O = 2.305 (2)-2.609 (2) A], while the other two are five-coordinate via one 1,4-butanediol [2.289 (2) and 2.349 (3) A] and four diolate O atoms [2.295 (2)-2.408 (2) A]. Corresponding to this, there are three- and four-coordinate diolate O atoms, the latter bridging Na atoms. The 1,4-butanediol molecules lie on inversion centres. The boron stereochemistry shows minor local perturbations from its usual tetrahedral state [B-O = 1.457 (4)-1.503 (4) A]. The resulting polymer packs as sheets parallel to the (10-1) plane crosslinked by the butanediol molecules. The structure was solved using data from a multiple crystal. PMID- 15750230 TI - Bis(tetra-n-butylammonium) and bis(tetraphenylphosphonium) salts of tris(2-oxo 1,3-dithiole-4,5-dithiolato)stannate(IV), both at 120 K. AB - The title compounds, (C16H36N)2[Sn(C3OS4)3], (I), and (C24H20P)2[Sn(C3OS4)3], (II), are examples of complex salts of the general form [Q]2[Sn(dmio)3], where Q is nBu4N+ or Ph4P+ and dmio is the 2-oxo-1,3-dithiole-4,5-dithiolate dianion. Features of both structures are the slightly distorted octahedral coordination of tin in the propeller-shaped dianions and the absence of any significant inter anion contacts. The structure of (I) is particularly notable because all of the dianions in the sample crystal have the same propeller configuration, which is very unusual in this type of structure. PMID- 15750231 TI - catena-Poly[[chlorodipyridinemanganese(II)]-mu3-6-oxo-1,6-dihydropyridine-2 carboxylato]. AB - The title one-dimensional chain polymer complex, [Mn(C6H4NO3)Cl(C6H5N)2]n, was isolated from the reaction of MnCl2 with 6-oxo-1,6-dihydropyridine-2-carboxylic acid (HpicOH) in pyridine. The asymmetric unit contains one [Mn(HPicO)Cl(py)2] moiety (py is pyridine), with the (HpicO)- ligand acting in a tridentate manner via the two carboxylate O atoms and the pyridone O atom. The operation of inversion centres generates eight- and 14-membered rings and, in conjunction with an a-axis translation, leads to an infinite chain extending along [100]. The Mn...Mn separations in this chain are 5.1069 (6) and 7.1869 (6) A. The MnII atom has a distorted octahedral coordination, with trans-axial pyridine ligands and with three O atoms and the Cl atom in the equatorial plane. The conformation of the 14-membered ring is stabilized by pairs of inversion-related N-H...O hydrogen bonds. PMID- 15750232 TI - Tris(2,2'-bioxazoline-kappa2N,N')copper(II) diperchlorate and tris(2,2' bioxazoline-kappa2N,N')nickel(II) diperchlorate. AB - In the two isomorphous title compounds, viz. tris[2,2'-bi(4,5-dihydro-1,3 oxazole)-kappa(2)N,N']copper(II) diperchlorate, [Cu(C6H8N2O2)3](ClO4)2, (I), and tris[2,2'-bi(4,5-dihydro-1,3-oxazole)-kappa2N,N']nickel(II) diperchlorate, [Ni(C6H8N2O2)3](ClO4)2, (II), the MII ions each have a distorted octahedral coordination geometry formed via six N atoms from three 2,2'-bioxazoline ligands. For each ligand, the two five-membered rings are nearly coplanar. It is noteworthy that the Jahn-Teller effect is stronger in (I) than in (II). The three dimensional supramolecular structures of (I) and (II) are formed via weak hydrogen-bonding interactions between O atoms from perchlorate anions and H atoms from 2,2'-bioxazoline ligands. PMID- 15750233 TI - Tetrakis(1-ethyl-1H-1,2,4-triazole-kappaN4)bis(nitrato-kappaO)copper(II) and bis(nitrato-kappaO)tetrakis(1-propyl-1H-1,2,4-triazole-kappaN4)copper(II). AB - The copper(II) environments for tetrakis(1-ethyl-1,2,4 triazole)dinitratocopper(II), [Cu(NO3)2(C4H7N3)4], and tetrakis(1-propyl-1,2,4 triazole)dinitratocopper(II), [Cu(NO3)2(C5H9N3)4], are distorted square bipyramidal. Both structures are centrosymmetric, with the copper(II) ions located at inversion centers coordinated by four N atoms of four triazole molecules and by two O atoms of two nitrate ions in an elongated octahedral geometry. This elongation is a result of the Jahn-Teller effect. The largest distortion is that of the N-Cu-O angles, which differ from 90 degrees by 5.68 (10) degrees in the ethyl and 5.59 (8) degrees in the propyl derivative. PMID- 15750234 TI - Form III of 2,2',4,4',6,6'-hexanitroazobenzene (HNAB-III). AB - The crystal structure of form III of the title compound, HNAB [systematic name: bis(2,4,6-trinitrophenyl)diazene], C12H4N8O12, has finally been solved as a pseudo-merohedral twin (monoclinic space group P2(1), rather than the orthorhombic space group C222(1) suggested by diffraction symmetry) using a dual space recycling method. The significant differences in the room-temperature densities of the three crystalline forms allow examination of molecular differences due to packing arrangements. An interesting relationship with the stilbene analog, HNS, is discussed. Interatomic separations are compared with other explosives and/or nitro-containing compounds. PMID- 15750235 TI - 17-Oxo-5alpha-androstane-3alpha,4beta-diyl diacetate and 17-oxo-5beta-androstane 3alpha,4beta-diyl diacetate. AB - The title compounds, both C23H34O5, are the 5alpha and 5beta configurations of two diacetate epimers. The 5beta-diacetate crystallizes in an hexagonal structure, unusual for steroid molecules. The unit cell has an accessible solvent volume of 358 A(3), responsible for clathrate behaviour. The 5beta-epimer also features some shorter than average bond lengths in the 3alpha,4beta-acetoxy groups. The conformations of the molecules of both epimers are compared with those obtained through ab initio quantum chemistry calculations. Cohesion of the crystals can be attributed to van der Waals and weak molecular C-H...O interactions. PMID- 15750236 TI - Polymorphism in ammonium 2,4,6-trimethylbenzenesulfonate. AB - During investigations into sulfide- and selenide-amination reactions using the aminating agent o-mesitylsulfonylhydroxylamine, the monoclinic, (I), and orthorhombic, (II), polymorphs of ammonium 2,4,6-trimethylbenzenesulfonate, NH4+.C9H11O3S-, have been crystallized. Investigation of the hydrogen-bonding motifs within the two polymorphs shows that both contain N+-H...O- hydrogen bonds between the ammonium cations and the 2,4,6-trimethylbenzenesulfonate anions. Polymorph (I) contains R(4)(4)(12) and R(4)(2)(8) graph-set ring motifs, while polymorph (II) contains the same R(4)(4)(12) ring motif in combination with an R(4)(3)(10) motif. The two hydrogen-bonding patterns result in slightly different packing structures for the two polymorphs, but both are based on a thick-sheet arrangement, in which the NH4+ cations are enveloped between two layers of 2,4,6 trimethylbenzenesulfonate anions. In (I), the aromatic rings of the anions are approximately coplanar, giving parallel sheets, whereas in (II) the sheets are antiparallel and the anions pack in a herring-bone manner within the sheets, with angles of 78.76 (8) degrees between the planes of the aromatic rings. PMID- 15750237 TI - Hydrogen-bonding patterns of two dihydroxylactone derivatives. AB - In the hydrogen-bonding networks of 8-hydroxy-5-hydroxymethyl-3,6 dioxatricyclo[6.3.1.0(1.5)]dodecan-2-one and 5,7-bis(hydroxymethyl)-3,6 dioxatricyclo[5.3.1.0(1.5)]undecan-2-one, both C11H16O5, layers and double strands, respectively, lead to the formation of chains connected by hydroxy-to hydroxy contacts, where the hydroxymethyl group, present in both structures, acts as a donor. The secondary structures differ in the hydrogen bonding of these chains via the second hydroxy group, which is involved in hydroxy-to-carbonyl and hydroxy-to-hydroxy bonds, respectively. PMID- 15750238 TI - 4'-Octyloxybiphenyl-4-carbonitrile polymorph III. AB - The title compound, C21H25NO, is a member of a well known family of liquid crystals (4-oxy-4'-cyanobiphenyls, OCBs) and packs in lamellar-type bilayers in the solid state, through CN...H hydrogen bonds. This packing type is analogous to that found of other members of the n-OCB homologous series, viz. 7-OCB and 9-OCB. PMID- 15750239 TI - 2-Iodo-6-methoxy-4-nitroaniline: tripartite ribbons built from N-H...O hydrogen bonds and iodo-nitro interactions are pi-stacked into sheets. AB - Molecules of the title compound, C7H7IN2O3, are linked by pairs of N-H...O hydrogen bonds into C(8)C(8)[R(2)(2)(6)] chains of rings, and antiparallel pairs of such chains are linked by a two-centre iodo-nitro interaction into tripartite ribbons. A single aromatic pi-pi stacking interaction links the ribbons into sheets. PMID- 15750240 TI - 3-Acetoxy-2-(acetylamino)pyridinium-1-squarate. AB - The title compound, alternatively known as 3-acetoxy-2-(acetylamino)pyridinium betaine of squaric acid, C13H10N2O6, has been synthesized. The bond distances within the squarate ring indicate two possible resonance structures. The mean planes of the pyridinium and squarate systems are inclined at an angle of 24.0 (2) degrees with respect to one another due to a strong intramolecular hydrogen bonding interaction between the amide NH group and a squarate O atom. In the extended structure, there are additional weak pi-pi and pi-ring interactions, which also stabilize the crystal structure. PMID- 15750241 TI - 7-Deaza-2'-deoxyguanosine. AB - In the title compound, 2-amino-7-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,7 dihydropyrrolo[2,3-d]pyrimidin-4-one, C11H14N4O4, the N-glycosylic bond torsion angle, chi, is anti [-106.5 (3) degrees ]. The 2'-deoxyribofuranosyl moiety adopts the 3T4 (N-type) conformation, with P = 39.1 degrees and tau(m) = 40.3 degrees . The conformation around the exocyclic C-C bond is ap (trans), with a torsion angle, gamma, of -173.8 (3) degrees . The nucleoside forms a hydrogen bonded network, leading to a close-packed multiple-layer structure with a head-to head arrangement of the bases. The nucleobase interplanar O=C-C...NH(2) distance is 3.441 (1) A. PMID- 15750242 TI - 7-Carboxylato-8-hydroxy-2-methylquinolinium monohydrate and 7-carboxy-8-hydroxy-2 methylquinolinium chloride monohydrate at 100 K. AB - Both 7-carboxylato-8-hydroxy-2-methylquinolinium monohydrate, C11H9NO3.H2O, (I), and 7-carboxy-8-hydroxy-2-methylquinolinium chloride monohydrate, C11H10NO3+.Cl .H2O, (II), crystallize in the centrosymmetric P-1 space group. Both compounds display an intramolecular O-H...O hydrogen bond involving the hydroxy group; this hydrogen bond is stronger in (I) due to its zwitterionic character [O...O = 2.4449 (11) A in (I) and 2.5881 (12) A in (II)]. In both crystal structures, the HN+ group participates in the stabilization of the structure via intermolecular hydrogen bonds with water molecules [N...O = 2.7450 (12) A in (I) and 2.8025 (14) A in (II)]. In compound (II), a hydrogen-bond network connects the Cl- anion to the carboxylic acid group [Cl...O = 2.9641 (11) A] and to two water molecules [Cl...O = 3.1485 (10) and 3.2744 (10) A]. PMID- 15750244 TI - Similar environments for the ClO4-, HSO4- and H2PO4- anions offered by strychnine self-assemblies. AB - In strychninium chlorate(VII) monohydrate, C21H23N2O2+.ClO4-.H2O, strychninium hydrogensulfate(VI) dihydrate, C21H23N2O2+.HSO4-.2H2O, and strychninium dihydrogenphosphate(V) dihydrate, C21H23N2O2+.H2PO4-.2H2O, the strychninium cations form bilayer sheets separated by water-anion sheets. The strychnine bilayer sheets in the three compounds are similar to one another. In all three structures, the surfaces of the cation and water-anion sheets exhibit donor acceptor matching. PMID- 15750243 TI - Hydrogen-bonding and C-H...pi interactions in 7-hydroxy-3-methoxy-4-methyl 5,6,7,8-tetrahydropyrido[1,2-c]pyrimidin-1(9H)-one. AB - In the title compound, C10H14N2O3, a pyrimidine ring is fused with a piperidine ring. The pyrimidine ring is planar, whereas the piperidine ring adopts a half chair conformation. The molecules of the title compound are connected via O-H...O intermolecular hydrogen bonds into infinite zigzag chains. The pyrimidine ring is involved in three C-H...pi interactions, which link the hydrogen-bonded chains into a three-dimensional framework. PMID- 15750245 TI - Tetramethylammonium 2,5-dinitrophenolate monohydrate. AB - The crystal structure of the title compound, C4H12N+.C6H3N2O5-.H2O, is built up from tetramethylammonium cations, 2,5-dinitrophenolate anions and water molecules. The nitro groups are almost coplanar with the aryl ring, which exhibits significant distortion from an ideal hexagonal form. The X-ray geometry of the tetramethylammonium cation shows slight distortion from the tetrahedral symmetry predicted by molecular orbital calculations. The O-H...O hydrogen-bonded water and 2,5-dinitrophenolate units are related by an inversion center and form dimers. The 2,5-dinitrophenolate anions, related by an inversion center and translation, are stacked to form a column along the [100] direction. PMID- 15750246 TI - Diethyl [(Z)-1-iodo-2-phenyl-1-hexenyl]phosphonate. AB - The crystal structure of the title compound, C16H24IO3P, is composed of independent molecules, in which one of the phosphonate ethoxy groups is disordered over two positions. The other ethoxy group lies sandwiched between two phenyl rings and, as a result, is ordered. This constrained environment also leads to two C-H...centroid interactions, one intra- and one intermolecular. Examination of the extended structure reveals the presence of chains of molecules held together by I...O interactions. PMID- 15750247 TI - Tolterodinium (+)-(2R,3R)-hydrogen tartrate. AB - In the crystal structure of (R)-N,N-diisopropyl-3-(2-hydroxy-5-methylphenyl)-3 phenylpropylaminium (2R,3R)-hydrogen tartrate, C22H32NO+.C4H5O6-, the hydrogen tartrate anions are linked by O-H...O hydrogen bonds to form helical chains built from R(2)(2)(9) rings. These chains are linked by the tolterodine molecules via N H...O and O-H...O hydrogen bonds to form separate sheets parallel to the (101) plane. PMID- 15750248 TI - Cyclo(L-leucyl-alpha,beta-dehydrophenylalanine): the first diketopiperazine containing an alpha,beta-dehydrophenylalanine residue. AB - The title compound (systematic name: 3-benzylidene-6-isobutylpiperazine-2,5 dione), C15H18N2O2, an alpha,beta-dehydrophenylalanine containing diketopiperazine, crystallizes in the space group P1 with two molecules in the asymmetric unit arranged antiparallel to one another. The alpha,beta dehydrophenylalanine (DeltaPhe) residue in this cyclic peptide retains its planarity but deviates from the standard conformations observed in its linear analogues. Each type of molecule forms a linear chain with molecules of the same type via pairwise N-H...O hydrogen bonds, while weaker C-H...O interactions link the chains together to form a three-dimensional network. PMID- 15750249 TI - C-halogen...pi dimer and C-H...pi interactions in 1-(2-bromo-4,5-dimethoxybenzyl) 2-butyl-4-chloro-1H-imidazole-5-carbaldehyde and 2-butyl-4-chloro-1-(6-methyl-1,3 benzodioxol-5-ylmethyl)-1H-imidazole-5-carbaldehyde. AB - The structures of the title compounds, C17H20BrClN2O3, (I), and C17H19ClN2O3, (II), are stabilized by intramolecular C-H...O and C-H...pi interactions. The stability of the molecular packing in (I) and (II) arises from a diverse set of weak intermolecular C-H...O, C-H...pi and C-halogen...pi interactions. In the crystal structure of (I), molecules aggregate in dimeric subunits via C-Br...pi interactions. The dimers are interlinked by C-H...O hydrogen bonds. The halogens cluster together and form a channel along the b axis. In (II), the packing is mainly governed by intermolecular C-H...O and C-H...pi interactions. PMID- 15750251 TI - 4-Nitrophenyl phenyl ether: sheets built from C-H...O and C-H...pi(arene) hydrogen bonds. AB - In the title compound, C12H9NO3, the ether C-O-C angle is 119.65 (10) degrees . The molecules are linked into sheets by a combination of C-H...O and C H...pi(arene) hydrogen bonds, reinforced by an aromatic pi-pi stacking interaction. PMID- 15750250 TI - The hydrogen-bonding network in (+)-N-tosyl-L-glutamic acid. AB - The asymmetric unit of the alpha polymorph of (+)-N-tosyl-L-glutamic acid, C12H15NO6S, contains two independent molecules which differ in conformation. The carboxylic acid groups form an infinite zigzag chain with characteristic R(2)(2)(8) rings running along the b axis. Intermolecular N-H...O and C-H...O contacts mediate the formation of a three-dimensional supramolecular structure described by R(4)(3)(22), R(6)(6)(44) and R(8)(8)(54) graph-set descriptors. Comparison of the extended structure with that of N-(benzenesulfonyl)glutamic acid shows that a subtle difference in the periphery of the molecule, i.e. the replacement of the toluyl group with a phenyl group, can be accompanied by dramatic changes in molecular assembly. PMID- 15750252 TI - A new phase of 7,16-dibenzyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane, and 7,16-dibenzyl-1,4,10,13-tetraoxa-7,16-diazoniacyclooctadecane bis(tetrafluoroborate) monohydrate, both determined at 123 K. AB - 7,16-Dibenzyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane, C26H38N2O4, (I), crystallizes in space group P2(1)/c, with two independent molecules adopting different conformations. The 'free' crowns adopt a typical 'arallelogram' shape, in which two methylene groups are turned inward toward the center of the ring and the benzyl groups splay out from the ring. In 7,16-dibenzyl-1,4,10,13-tetraoxa 7,16-diazoniacyclooctadecane bis(tetrafluoroborate) monohydrate, C26H40N2O4(2+).2BF4-.H2O, (II), the macrocycle is centrosymmetric, and the protonated N atoms adopt an endo-endo orientation that is stabilized by a bifurcated N-H...O hydrogen bond, where the O atoms of the macrocycle act as hydrogen-bond acceptors. The phenyl groups of the benzyl side arms are turned above and below the macrocycle; C-H...pi interactions between the phenyl substituents and two macrocyclic methylene H atoms govern the overall conformation of the macrocycle. Bridging tetrafluoroborate anions link the macrocyclic cations via weak C-H...F hydrogen bonds into channels running along [100], which are filled by the weakly hydrogen-bonded water molecules. PMID- 15750253 TI - 1,4-Bis(p-tolylethynyl)benzene. AB - The rod-like molecule of the title hydrocarbon, C24H18, is centrosymmetric, with the centroid of the central benzene ring residing on an inversion center. The molecules display a planar conformation of the benzene rings and aggregate into stacks along the [010] direction via Csp(3)-H...pi(arene) interactions, thus forming a stair-like pseudo-two-dimensional network. Each molecule acts as both a C-H hydrogen donor and a pi-arene acceptor, forming four hydrogen bonds per molecule. PMID- 15750254 TI - Stress profile in essential hypertension. AB - This study was undertaken to test the hypothesis that basal sympathetic activity and sympathetic reactivity to stress are increased in patients with essential hypertension. One hundred and fifteen randomly selected patients with essential hypertension and an equal number of age- and sex-matched normal controls were included in this study. Various parameters, viz., heart rate, respiratory rate, blood pressure, peripheral skin temperature, electromyographic activity of the frontalis muscle, skin conductance (measured by electrodermography), and blood pressure, were measured in the resting state in both groups. These parameters were then measured during the performance of various stressful activities, such as mental arithmetical calculations, thinking of unpleasant thoughts, staring at a fixed point, catching of a dropped object, and reading aloud for 1 min each, and again over a 1-min quiet period following each stressful activity. Changes in various parameters in the two groups were analyzed and compared. Baseline heart rate, respiratory rate, electromyographic activity and peripheral skin temperature were higher-while skin conductance was lower-in the hypertensive group than in the control group. The increases in systolic blood pressure, diastolic blood pressure, heart rate, respiratory rate, electromyographic activity, and peripheral skin temperature in response to stress were greater in hypertensive patients than in controls, while skin conductance showed a higher elevation in controls than in hypertensive patients. In conclusion, sympathetic nervous system activity is increased in patients with essential hypertension. The reduced skin conductance and high peripheral skin temperature in these patients may be due to a possible dermal neurovascular dysfunction. PMID- 15750255 TI - Arterial stiffening as a possible risk factor for both atherosclerosis and diastolic heart failure. AB - While arterial stiffness is known to be related to atherosclerosis, the association between arterial stiffness and cardiac systolic and diastolic function in hypertension has not been fully evaluated. The present study was conducted to simultaneously evaluate the relationship of brachial-ankle pulse wave velocity (PWV) to parameters reflecting atherosclerosis and to those reflecting the risk of congestive heart failure in patients with hypertension. In 147 patients with hypertension, the left ventricular ejection fraction, the ratio of the peak velocity of early rapid filling and the peak velocity of atrial filling (E/A ratio), and left ventricular mass index were obtained from echocardiographs, the intima-media thickness of the common carotid artery was obtained by ultrasonography, the plasma B-type natriuretic peptide (BNP) level was measured by radioimmunoassay, and the brachial-ankle PWV was measured by the volume rendering method. Brachial-ankle PWV correlated positively with the intima media thickness of the carotid artery, E/A ratio and BNP. Multiple linear regression analysis demonstrated that the relationship between the brachial-ankle PWV and the E/A ratio was significantly independent from other clinical variables. The receiver operator characteristic curve demonstrated that a brachial-ankle PWV of 1,600 cm/s was useful to discriminate mild cardiac diastolic dysfunction (E/A ratio of < or =0.75) (sensitivity=78% and specificity=58%). The present study demonstrated that increased brachial-ankle PWV relates not only to the parameters reflecting atherosclerosis but also to those reflecting cardiac diastolic dysfunction. Therefore, increased arterial stiffness is a possible simultaneous risk for atherosclerotic cardiovascular disease and diastolic heart failure in patients with hypertension. PMID- 15750256 TI - Home blood pressure is a better predictor of carotid atherosclerosis than office blood pressure in community-dwelling subjects. AB - An increase in the thickness of the intima-media (IMT) of the carotid artery is associated with an increased risk of cardiovascular morbidity and mortality. Hypertension is one of the underlying mechanisms for the progression of carotid atherosclerosis. However, office blood pressure (BP) has been shown to have only a weak association with carotid IMT. Since self-measured home BP has less variation than office BP, home BP could be a better predictor of carotid atherosclerosis. To explore this hypothesis, we compared the relationships between carotid IMT and office BP or home BP in a community-dwelling population. One-hundred and one community residents, aged 50 years or older and not taking any medication, were enrolled in this study. Morning home BP was measured according to the guidelines of the Japanese Society of Hypertension. The results were recorded for 2 weeks and averaged. Carotid atherosclerosis was defined as IMT > or =0.80 mm, which corresponds to the first quartile. Home systolic BP showed a more significant association with carotid IMT (r=0.422, p <0.0001) than with office systolic BP (r=0.021, p=0.027). Logistic regression analysis for the presence of carotid atherosclerosis further showed that the relative risk of hypertension defined using home BP (> or =135/85 mmHg) was 6.3 (95% confidence interval [CI]: 2.0 to 19.6), while that using office BP was 1.5 (95% CI: 0.5 to 4.2). These results suggest that home BP is a better predictor of the development of carotid atherosclerosis than office BP. PMID- 15750257 TI - Relation of pulsatility of brachial artery pressure to resistant hypertension. AB - Few studies have examined predictors of resistant hypertension. The aim of this study was to observe the relationship between resistant hypertension and the pulsatility of the brachial artery pressure, which is characterized as pulse pressure/diastolic pressure (PP/DP) and is a simple index of aortic input impedance. We obtained home blood pressure (BP) measurements for 102 patients aged 40-75 years with either office systolic BP (SBP) > or =140 mmHg or office diastolic BP (DBP) > or =90 mmHg. Patients were given a single antihypertensive agent or left untreated during the 2-week baseline period. Thereafter, patients were treated with 1 to 3 antihypertensive drugs for 1 year with a goal of achieving a home BP of less than 135/85 mmHg. At follow-up, 72 patients were taking a single drug with good BP control, 21 were taking two drugs with good BP control, and 9 were taking three drugs with poor BP control. Although office SBP at baseline was similar among the three groups, home morning and evening SBP at baseline in the single drug group were lower than those of the two- or three-drug groups (p <0.01). Although office PP/DP at baseline did not differ among the three groups, home morning and evening PP/DP at baseline were highest in the three-drug group (p <0.01). In multivariate analysis, only mean home PP/DP at baseline was correlated with BP control. There is a correlation between the pulsatility of the brachial artery pressure and the degree of BP control. PMID- 15750258 TI - Ambulatory blood pressure level rather than dipper/nondipper status predicts vascular events in type 2 diabetic subjects. AB - To clarify which parameter, diurnal pattern of blood pressure (BP) or level of BP variability, has the stronger predictive value for fatal and nonfatal vascular events, vital status after a mean (+/-SD) follow-up period of 86+/-46 months was determined in 392 type 2 diabetic subjects without any history of vascular disease, in whom the 24-h BP profile had been monitored between 1988 and 1998. After the exclusion of 28 subjects who died during the follow-up period of causes unrelated to diabetes, 364 subjects were recruited for further analysis. A total of 147 first events, including 50 fatal vascular events and 97 nonfatal vascular events, were recorded during the follow-up period. The rates of various vascular events increased with both reduced nocturnal falls in systolic BP (SBP) and levels of all ambulatory BP parameters. The ambulatory BP parameter showing the largest area under the receiver operating characteristic curve (ROCAUC) for fatal events was the mean 24-h pulse pressure (PP), and that for nonfatal events was the mean nighttime SBP; both exceeded the respective values of nocturnal fall in SBP. Furthermore, when dipper and nondipper diabetic subjects were divided into subgroups based on the 24-h PP (54.3 mmHg) and the nighttime SBP (116.5 mmHg) cut off points derived from the ROC analyses, Kaplan-Meier plots showed that the diabetic subgroups with high ambulatory BP levels had worse outcomes, independent of dipper/nondipper status. Finally, these parameters were applied to the Cox model with the values of nocturnal fall in SBP and other confounding factors, and results showed that mean 24-h PP and mean nighttime SBP predicted fatal and nonfatal vascular events, respectively, more strongly than nocturnal fall in SBP in type 2 diabetic subjects. These findings therefore suggest that ambulatory BP levels in type 2 diabetic subjects have a higher predictive value for organ damage and death compared with diurnal BP patterns or dipper/nondipper status. PMID- 15750259 TI - valsartan in elderly isolated systolic hypertension (VALISH) study: rationale and design. AB - Although antihypertensive therapy has been proven to reduce cardiovascular morbidity and mortality, it is unclear how much blood pressure should be decreased in elderly patients with hypertension. The Valsartan in Elderly Isolated Systolic Hypertension (VALISH) study is a multicenter parallel-group study comparing the incidence of cardiovascular events between two target systolic blood pressure levels, below 140 mmHg and below 150 mmHg, under treatment with valsartan, an angiotensin II receptor blocker, as an initial antihypertensive drug in elderly patients with isolated systolic hypertension. The number of patients to be recruited is 3,000 and the duration of follow-up is at least 2 years. This 3,000-patient trial was designed with a two-sided alpha level of 0.05 and 80% power to detect the difference in incidence of cardiovascular events between the target blood pressure levels based on estimation of the cardiovascular events ratio as 21.5/1,000 patient-years and 29.1/1,000 patient-years for the two blood pressure levels. The VALISH study, a large-scale investigator-initiated trial in Japan, will determine whether age should be considered in setting target blood pressure in treatment of isolated systolic hypertension in elderly patients. PMID- 15750260 TI - The methylentetrahydrofolate reductase gene variant (C677T) as a risk factor for essential hypertension in Caucasians. AB - Essential hypertension (EH) is a common, multifactorial disorder likely to be influenced by multiple genes of modest effect. The methylenetetrahydrofolate reductase (MTHFR) gene C677T mutation is functionally important, being strongly associated with reduced enzyme activity and increased plasma levels of homocysteine. Mild hyperhomocysteinemia is a known risk factor for cardiovascular disease (CVD) and hypothesised also to be involved in hypertension pathophysiology. The present study was performed to determine the prevalence of the 677T mutation in Australian Caucasian patients diagnosed with EH and to test whether the C677T variant is associated with the disorder. A case-control cohort, consisting of 250 EH patients and 250 age, sex and racially matched normotensive controls, were used for the association study. Comparison of C677T allele frequencies revealed a higher proportion of the mutant allele (T) in the EH group (40%) compared to unaffected controls (34%) (p=0.07). Furthermore, genotypic results indicated that the prevalence of the homozygous mutant genotype (T/T) in the affected group was higher than that of controls (14%:10%) (p=0.17). Interestingly, conditional logistic regression showed that the MTHFR C677T mutation conferred a mild, yet significant increase in risk of essential hypertension after adjusting for body mass index (odds ratio=1.57, 95% confidence interval: 1.04-2.37, p=0.03). These findings require further investigation in large independent samples, but suggest that essential hypertension, like CVD, may be mildly influenced by the MTHFR C677T variant. PMID- 15750261 TI - Excessive alcohol intake increases the risk of arterial stiffening in men with normal blood pressure. AB - Excessive alcohol intake is a known atherosclerotic cardiovascular risk factor. However, the relation between excessive alcohol intake and atherosclerotic cardiovascular risk in subjects with normal blood pressure has not been fully elucidated. This cross-sectional study investigated the relationship between alcohol intake and arterial stiffness, as assessed using brachial-ankle pulse wave velocity, in men with normal blood pressure. Middle-aged male workers who were found to have a normal blood pressure during an annual health check-up performed in May-July 2000 (n=1,682) were enrolled in the study. The subjects' laboratory data were analyzed, and information on drinking and smoking habits was obtained by a self-administered questionnaire. The brachial-ankle pulse wave velocity was measured using volume plethysmography. The mean brachial-ankle pulse wave velocities of men with an alcohol intake of 40-59 g/day and > or =60 g/day were larger than those of non-drinkers and men with an alcohol intake of <20 g/day. A multivariate general linear model was used to identify the association between alcohol intake and brachial-ankle pulse wave velocity after controlling for other risk factors using a multivariate analysis. An alcohol intake of more than 60 g of ethanol/day was significantly associated with an elevated brachial ankle pulse wave velocity, after controlling for conventional atherosclerotic cardiovascular risk factors. In conclusion, the present results suggest that excessive alcohol intake increases the atherosclerotic cardiovascular risk associated with arterial stiffening in men with normal blood pressure. PMID- 15750262 TI - Genistein inhibits expressions of NADPH oxidase p22phox and angiotensin II type 1 receptor in aortic endothelial cells from stroke-prone spontaneously hypertensive rats. AB - Phytoestrogens are considered to be natural selective estrogen receptor modulators exerting antioxidant activity and improving vascular function. However, the mechanisms responsible for their antioxidative effects remain largely unknown. This study tested the hypothesis that genistein may provide significant endothelial protection by antioxidative effects through attenuating NADPH oxidase expression and activity. The results showed that genistein suppressed the expressions of the p22phox NADPH oxidase subunit and angiotensin II (Ang II) type 1 (AT1) receptor in a concentration- and time-dependent manner in aortic endothelial cells from stroke-prone spontaneously hypertensive rats examined by Western blot analysis. Treatment with genistein also remarkably reduced the Ang II-induced superoxide by the reduction of nitroblue tetrazolium, inhibited nitrotyrosine formation, and attenuated endothelin-1 production by ELISA via the stimulation of Ang II. However, when cells were pretreated with ICI 182780, an estrogen-receptor antagonist, at a concentration of 50 micromol/l for 30 min and then co-incubated with ICI-182780 and genistein for 24 h, the inhibitory effect of genistein was not blocked. In contrast, the inhibitory effect of genistein treatment was partially reversed by 30-min pretreatment of endothelial cells with GW9662, a peroxisome proliferator-activated receptor gamma (PPARgamma) antagonist. Genistein thus appears to act as an antioxidant at the transcription level by the downregulation of p22phox and AT1 receptor expression. Our data also showed that the PPARgamma pathway was involved, at least in part, in the inhibitory effect of genistein on the expression of p22phox and AT1 receptors. The endothelial-protective effects of phytoestrogen may contribute to improvement of cardiovascular functions. PMID- 15750263 TI - Effects of angiotensin II type 1 receptor antagonist on smooth muscle cell phenotype in intramyocardial arteries from spontaneously hypertensive rats. AB - To clarify the precise mechanisms involved in the reduced coronary flow reserve in hypertension, we compared the effects of the angiotensin II type 1 (AT1) receptor antagonist FK-739 with those of the angiotensin-converting enzyme (ACE) inhibitor enalapril for 6 weeks on the smooth muscle (SM) cell phenotype in intramyocardial arteries from male Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Compared with WKY, SHR showed a significant increase in left ventricular (LV) hypertrophy and fibrosis, blood pressure (BP), and vascular remodeling of the intramyocardial arteries, and a significant decrease in endothelial NO synthase and the contractile-type myosin heavy chain isoform SM2 of the intramyocardial arteries as well as calponin 1 and GATA-6. In the hearts of SHR, both drugs equivalently and significantly reduced BP, which was still significantly higher than that in the WKY groups, and also reduced LV hypertrophy and fibrosis, whereas endothelial NO synthase was significantly restored. Although both drugs showed little effect on the vascular remodeling of the intramyocardial arteries in the SHR hearts, FK-739, but not enalapril, significantly restored SM2 and GATA-6 in the SHR hearts to the same levels as those of the vehicle WKY group. The effects of the two drugs on these indices were not observed in the three WKY hearts. Thus, the AT1 receptor antagonist may modulate the SM cell phenotype toward the contractile-type more effectively than the ACE inhibitor before the morphological changes occur in the intramyocardial arteries of the SHR hearts. PMID- 15750264 TI - Endoscopic submucosal dissection (ESD) is being accepted as a new procedure of endoscopic treatment of early gastric cancer. PMID- 15750265 TI - Mitochondrial DNA mutation and functional dyspepsia. PMID- 15750266 TI - Clinical significance of cyclophosphamide-induced cardiotoxicity. PMID- 15750267 TI - Endothelial function and coronary spastic angina. AB - Coronary spasm plays an important role in the pathogenesis of not only variant angina but also coronary heart disease in general including acute coronary syndromes, especially in the Japanese population. The vascular endothelium has been reported to be a multifunctional organ whose integrity is essential for normal vascular physiology. Vascular endothelial dysfunction can be a critical factor in the pathogenesis of ischemic heart disease. Acetylcholine and methacholine cause vasodilation by endothelium-derived relaxing factor when the endothelium is functioning normally, whereas they cause vasoconstriction when the endothelium is removed or damaged. Coronary spasm can be induced by a variety of stimuli with different mechanisms of action, including acetylcholine and methacholine. Patients with coronary spasm may have a disturbance in endothelial function as well as local hyperreactivity of the coronary arteries. PMID- 15750268 TI - A trial to assess the amount of insulin antibodies in diabetic patients by surface plasmon resonance. AB - OBJECTIVE: To measure the amount and affinity of insulin antibodies, we performed a trial to establish a new method for quantitative and qualitative analysis of these antibodies by using surface plasmon resonance (BIAcore system). METHODS: Real-time detection of insulin antibody interaction and kinetic analysis were performed using the BIAcore system. PATIENTS OR MATERIALS: Eight diabetic patients with insulin antibodies and whose fasting total immunoreactive insulin levels were more than 100 microU/ml were selected. The patients with and without recurrent hypoglycemia were classified into hypoglycemic episode-positive or hypoglycemic episode-negative groups, respectively. Seven diabetic patients without insulin antibodies were selected as controls. RESULTS: In the 8 patients, the concentration of insulin antibodies ranged from 2.91 to 16.3 microg/ml and insulin antibodies were not detected in the control group. The apparent KD (dissociation constant) and kd (the dissociation rate constant) values of the patients were much larger than those seen for the anti-human insulin monoclonal antibody. The KD values were significantly higher in the hypoglycemic episode positive group than in the hypoglycemic episode-negative group (p<0.05). No significant differences in the concentration, the ka (the association rate constant) and the kd values were noted between the groups. CONCLUSION: The data suggests that insulin antibodies of the patients have an apparently lower affinity status in sera as compared with that for the anti-human insulin monoclonal antibody, and dissociate easily from the immune-complex in the sera, especially in cases where there is recurrent hypoglycemia in the patients. Therefore insulin antibody characteristics are one of the causative factors in hypoglycemic episodes. PMID- 15750269 TI - Patients preferences in chemotherapy for advanced non-small-cell lung cancer. AB - OBJECTIVE: To determine how Japanese patients with lung cancer weigh potential survival, chemotherapy response rate, and symptom relief against the potential toxicity of different treatments in cancer chemotherapy. METHODS AND PATIENTS: We used a questionnaire describing a hypothetical situation about stage IV non-small cell lung cancer. Seventy-three patients with lung cancer who had received chemotherapy and 120 patients with other respiratory disease as the control group were asked to rate the minimal benefit that would make two hypothetical treatments acceptable. For "chance of cure," "response but not cure," and "symptom relief," the subjects could give answers from 1% to 100% and for prolonging life could give answers from 1 to 60 months. RESULTS: Patients with lung cancer were significantly more likely than were patients with other respiratory diseases to accept either intensive or less-intensive treatments for a potentially small benefit for "chance of cure," "response but not cure," and "symptom relief". The degree of survival advantage that patients require before accepting cancer treatment with its associated toxicity varied widely. If their lives were prolonged 3 months, 19% and 21% of patients with lung cancer would choose to receive intensive and less-intensive treatment, respectively. When the chance of symptom relief was 70%, 73% of patients with lung cancer were willing to choose intensive chemotherapy. Factor associated with patients' choice of chemotherapy in both groups was age. CONCLUSION: Oncologists must consider the substantial range of attitudes to chemotherapy among patients when making treatment decisions and they must give patients the opportunity to be included in this process. PMID- 15750270 TI - Malignant gastrointestinal stromal tumor of the small intestine complicated with pulmonary tuberculosis during treatment with imatinib mesylate. AB - We describe a patient who had a metastatic gastrointestinal stromal tumor (GIST) after previous failed extensive therapy, including multiple surgeries and hepatic artery embolization. Within a few months of starting administration of imatinib mesylate, the patient exhibited a clinical response with grade 3 neutropenia, when pulmonary tuberculosis developed. A c-kit mutation in exon 11 was detected only in metastatic liver specimens. It is unclear whether or not pulmonary tuberculosis may be induced by imatinib mesylate treatment, but caution is warranted in immunocompromised GIST patients. This is the first report of tuberculosis associated with neutropenia during imatinib mesylate treatment. PMID- 15750271 TI - Cyclophosphamide-induced cardiomyopathy in a patient with seminoma and a history of mediastinal irradiation. AB - A 17-year-old man with mediastinal seminoma was treated with chemotherapy and mediastinal irradiation therapy. Then he received high-dose chemotherapy containing cyclophosphamide (CY) followed by autologous peripheral blood stem cell transplantation. He suffered from CY-induced cardiomyopathy beginning six days after the administration of high-dose CY. The predictable factors associated with the onset of CY-induced cardiomyopathy are not precisely known. It is suggested that the history of mediastinal irradiation was responsible for the onset of cardiomyopathy. PMID- 15750272 TI - Sensorineural hearing loss combined with Takayasu's arteritis. AB - A 49-year-old woman complained of hearing loss and diminution of left radial arterial pulsation. She had been diagnosed with sudden deafness and treated with corticosteroids. Her audibility deteriorated again after the cessation of the therapy. Angiograms showed stenosis in the bilateral carotid arteries, the left vertebral artery, the left subclavian artery, and the pulmonary arteries. She was diagnosed with Takayasu's arteritis. After steroid therapy was restarted, there were improvements in her audibility, radial arterial pulsation, and levels of inflammatory markers (erythrocyte sedimentation rate, C-reactive protein, and gamma-globulin), fibrinogen, interleukin-6, and RANTES (regulated on activation, normal T cell expressed and secreted). PMID- 15750273 TI - Re-entry circuit in ventricular tachycardia due to focal fatty-fibrosis in a patient with myotonic dystrophy. AB - A 69-year-old man with a recurrent ventricular tachycardia (VT) was admitted. The patient was diagnosed as myotonic dystrophy type 1 (DM1) and DNA analysis revealed 1,800 CTG-repeat expansion in the myotonic dystrophy protein kinase (DMPK) gene. Ultrasonic cardiogram (UCG), left ventriculogram (LVG) and magnetic resonance imaging (MRI) did not show any abnormal sign including fatty infiltration. But, endomyocardial biopsy obtained from ventricular outflow tract revealed severe fatty infiltration and interstitial fibrosis. Radiofrequency catheter ablation at the biopsy site could eliminate VT, so it was strongly suggested that the re-entry circuit was formed by focal fatty-fibrosis. Careful observation should be continued for a long period. PMID- 15750274 TI - Dynamic electrocardiographic changes due to cardiac compression by a giant hiatal hernia. AB - A 79-year-old woman was admitted due to chest pain with T wave inversion and anasarca. Echocardiography demonstrated a mass compressing the heart and computed tomography revealed a giant hiatal hernia within the intrathoracic stomach located just behind the heart. After drainage of the gastric contents, the T wave inversion disappeared, but subsequent ST elevation in leads V1-V6 was noted. After surgical correction of the hiatal hernia, the ST segment elevation returned to a nearly normal level. The changes in the compressed heart induced by hiatal hernia may cause pericarditis resulting in electrocardiographic changes. PMID- 15750275 TI - Pulmonary coccidioidomycosis that formed a fungus ball with 8-years duration. AB - Coccidioidomycosis, caused by inhaling Coccidioides immitis, is a mycosis imported from endemic regions including the southwestern United States. C. immitis is so virulent that even a short-term stay in the endemic area can provide a chance for infection. Here, we report a 33-year-old Japanese man with formation of a fungus ball inside the pulmonary cavity secondary to coccidioidomycosis with a duration of 8 years, which is considered rare. He was infected with C. immitis in the United States in 1996. A nodule remained in the lung, which later cavitated with fungus ball formation. We identified Coccidioides immitis in the cultured specimen from the cavity and serum antibodies against it. We performed a lobectomy in 2003 since anti-fungal treatment was only temporarily effective. He is still free of disease 6 months later. PMID- 15750276 TI - Toxicity and outcome of intensive chemotherapy for acute lymphoblastic leukemia complicated with Turner's syndrome. AB - A 17-year-old woman was diagnosed as acute lymphoblastic leukemia (ALL). As she had chromosomal abnormalities of 44, XO, der(9)t(3;9)(q11;p13), der(10;19)(q10;p10), del(15)(q15), -16, -19, +22 with the presence of ovarian dysplasia and abnormal physical features, a diagnosis of Turner's syndrome was made. She received an induction chemotherapy, which consisted of daunorubicin, cyclophosphamide, vincristine, L-asparaginase and prednisolone. Although, severe liver dysfunction was observed, the patient achieved a complete remission (CR) on day 31 following chemotherapy and has maintained CR for more than five years. The recording of such cases may well be of value to clarify toxicity and outcome after chemotherapy for patients with ALL complicated with Turner's syndrome. PMID- 15750277 TI - Behcet disease presenting with neurological complications immediately after conversion from conventional cyclosporin A to microemulsion formulation. AB - A 53-year-old man with Behcet disease was treated with conventional cyclosporin A (CyA), because of refractory bilateral uveitis. Immediately following the conversion from conventional CyA to a microemulsion formulation, he presented with neurological complications. The neurological findings, pleocytosis of the cerebrospinal fluid (CSF) and brainstem lesions revealed by brain magnetic resonance imaging (MRI) suggested neuro-Behcet disease. After discontinuing CyA and introducing oral prednisolone, the neurological symptoms, pleocytosis of CSF and brainstem lesions on MRI improved. Although the microemulsion formulation, which can maintain a stable level of blood CyA, is a useful agent for the control of ocular lesions in Behcet disease, the resulting abrupt increase in blood CyA level may have induced neuro-Behcet disease. PMID- 15750278 TI - Lhermitte's sign in alcoholic myelopathy without portosystemic shunting: MRI evaluation. AB - We conducted spinal MR imaging on a 35-year-old man with Lhermitte's sign that had manifested over the previous 4 years. He had consumed more than 500 ml of whisky daily for at least 10 years. However, he did not show any evidence of severe liver disease with hepato-systemic blood shunting. Neurologic examination revealed markedly depressed sense of vibration in the feet and mild spasticity in the lower limbs, together with Lhermitte's sign. MR imaging revealed abnormal signal intensity in the posterior column spanning the whole length of the upper cervical cord, which is consistent with Lhermitte's sign. PMID- 15750279 TI - Hemorrhagic cerebral sinus thrombosis in a case of controlled ulcerative colitis. PMID- 15750280 TI - Massive hemoptysis due to pulmonary tuberculosis. PMID- 15750281 TI - Massive hemorrhage into the iliopsoas muscle. PMID- 15750283 TI - Clinical importance of miliary pattern in the chest X-ray of a patient with fever of unknown origin. PMID- 15750282 TI - Crowned dens syndrome. PMID- 15750284 TI - Effects of growth hormone on the differentiation of mouse B-lymphoid precursors. AB - Growth hormone (GH) has been known to enhance immune responses directly or through insulin-like growth factor-I (IGF-I). The present study aimed to clarify the roles of GH in the differentiation of B-lineage precursors. In short-term bone marrow cultures, which contained stem cells and early B-lineage cells, GH (10 mug/L) treatment for one day decreased the percentages of stem cells (0.5 fold) and increased those of B-lineage cells (1.4-fold). Furthermore, GH changed the expressions of transcription factors for B cell progenitors differentiation such as paired box gene-5 (Pax-5), immunoglobulin-associated-alpha (Ig alpha)/CD79a, Ig-beta/CD79b, and IGF-I. Thus, a physiological concentration of GH stimulated the differentiation of B-lymphoid precursors from bone marrow stem cells. Since mRNAs of both GH and GH receptor were present in stem cells and B cell precursors in bone marrow, GH may modulate B-lymphoid precursors development in an autocrine or paracrine manner in bone marrows. PMID- 15750285 TI - Changes in pH increase perfusion pressure of coronary arteries in the rat. AB - Stricture of coronary arteries is closely related to ischemic heart disease. The purpose of this study was to examine whether changes in pH caused contraction of rat coronary arteries, as determined using Langendorff perfused hearts. Changing the pH of the perfusate increased perfusion pressure as an indication of the contractile state of coronary arteries. Alkaline pH-induced increase of perfusion pressure in Wistar Kyoto rats (WKY) was almost identical to that of spontaneously hypertensive rats (SHR), whereas acidic pH-induced increase in SHR was much greater than that in WKY. Acidic pH-induced increase in perfusion pressure was inhibited by verapamil, cromakalim, and adenosine. Feeding WKY with N(G)-nitro-L arginine resulted in hypertension followed by enhanced acidic pH-induced increase in perfusion pressure. These results suggest that acidic-pH induced contraction of rat coronary arteries is caused by Ca(2+) influx through voltage-dependent Ca(2+) channels and the contraction is enhanced by hypertension. PMID- 15750286 TI - Effects of AF-DX116 and other muscarinic receptor antagonists on orthostatic hypotension in autonomic imbalanced (SART-stressed) rats. AB - SART (specific alternation of rhythm in temperature)-stressed rats are an animal model of autonomic imbalance created by exposing animals to repeated cold stress. The SART-stressed rats have been shown to easily develop orthostatic hypotension (OH). In this study, effects of AF-DX116, a selective M(2) antagonist, and other muscarinic receptor antagonists on OH were investigated in SART-stressed and unstressed rats. Each anesthetized rat was canulated into the left common carotid artery, and blood pressure (BP) and heart rate were measured. Stimulation for postural change was initiated by head-up tilting. As the indices of OH, the maximum fall of BP, % reflex (recovery from maximum fall), and the area enclosed between the baseline and the recovery curve for BP (AUC) were used. Large AUC and small % reflex in SART-stressed rats were changed, becoming similar to those of the unstressed rats by AF-DX116 and methoctoramine. Atropine and methylatropine had similar effects to AF-DX116. However, the effects of methoctoramine, atropine, and methylatropine were less than that of AF-DX116. Pirenzepine was not effective. In conclusion, it was suggested in SART-stressed rats that OH was related to hyperactivity in the parasympathetic nerve and the M(2) receptor played the major role in OH. PMID- 15750287 TI - CB(1) cannabinoid receptor stimulation modulates transient receptor potential vanilloid receptor 1 activities in calcium influx and substance P Release in cultured rat dorsal root ganglion cells. AB - Cannabinoids have been reported to have analgesic properties in animals of acute nociception or of inflammatory and neuropathic pain models, but the mechanisms by which they exert such alleviative effects are not yet fully understood. We investigated whether the CB(1)-cannabinoid-receptor agonist HU210 modulates the capsaicin-induced (45)Ca(2+) influx and substance P like-immunoreactivity (SPLI) release in cultured rat dorsal root ganglion (DRG) cells. HU210 attenuated the capsaicin-induced (45)Ca(2+) influx and this effect was reversed by the CB(1) antagonist AM251. Treatment of DRG cells with 100 nM bradykinin for 3 h potentiated capsaicin-induced SPLI release accompanied with the induction of cyclooxygenase-2 mRNA expression. The potentiation of SPLI release by bradykinin was reversed by HU210 or the protein kinase A (PKA) inhibitor H-89. HU210 also reduced forskolin-induced cyclic AMP production and forskolin-induced potentiation of SPLI release. These results suggest that CB(1) could inhibit either the capsaicin-induced Ca(2+) influx or the potentiation of capsaicin induced SPLI release by a long-term treatment with bradykinin through involvement of a cyclic-AMP-dependent PKA pathway. In conclusion, CB(1)-receptor stimulation modulates the activities of transient receptor potential vanilloid receptor 1 in cultured rat DRG cells. PMID- 15750288 TI - Simultaneous real-time detection of initiator- and effector-caspase activation by double fluorescence resonance energy transfer analysis. AB - Fluorescence resonance energy transfer (FRET) with green fluorescent protein (GFP) variants has become widely used for biochemical research. In order to expand the choice of fluorescent range in FRET analysis, we designed various color versions of the FRET-based probes for caspase activity, in which the substrate sequence of the caspase was sandwiched by donor and acceptor fluorescent proteins, and studied the potential of these color versions as fluorescent indicators. Six color versions were constructed by a combination of cyan fluorescent protein (CFP), GFP, yellow fluorescent protein (YFP), and DsRed. Real-time monitoring in single cells revealed that all probes could detect caspase activation during tumor necrosis factor (TNF)-alpha-induced cell death as a fluorescent change. GFP-DsRed and YFP-DsRed were as sensitive as CFP-YFP, and CFP-DsRed also showed a large fluorescent change. By using two probes, CFP-DsRed and YFP-DsRed, we carried out simultaneous multi-FRET analysis and revealed that the initiator- and effector-caspases were activated almost simultaneously in TNF alpha-induced cell death. These findings may give experimental bases for the development of novel techniques to analyze multi-events simultaneously in single cells by using FRET probes in combination. PMID- 15750289 TI - The role of sigma receptors in depression. AB - Behavioral models used to test potential antidepressants have shown that ligands that bind to sigma receptors possess "antidepressant-like" properties. The focus of this review is to discuss the literature concerning sigma receptors and their ligands, with respect to their antidepressants properties. In addition to the behavioral data, we discuss electrophysiological and biochemical models demonstrating sigma receptors' ability to modulate important factors in the pathophysiology of depression and/or the mechanisms of action of antidepressants such as the serotonergic neurotransmission in the dorsal raphe nucleus (DRN) and the glutamatergic transmission in the hippocampus. We also discuss the significance of these two systems in the mechanism of action of antidepressants. Sigma ligands have potential as antidepressant medications with a fast onset of action as they produce a rapid modulation of the serotonergic system in the DRN and the glutamatergic transmission in the hippocampus. As these effects of sigma ligands may produce antidepressant properties by completely novel mechanisms of action, they may provide an alternative to the antidepressants currently available and may prove to be beneficial for treatment-resistant depressed patients. PMID- 15750290 TI - Mitochondrial dysfunction, endoplasmic reticulum stress, and apoptosis in Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common neurodegenerative disorder of late life characterized by insidious, chronic, and progressive memory impairment in association with the accumulation of senile plaques, neurofibrillary tangles, and massive loss of neurons. Apoptosis is believed to be an important contributor to progression and pathology of neurodegeneration in AD. There is considerable evidence that amyloid beta-peptide, a major component of senile plaques, has the capacity to activate intracellular apoptosis pathways leading to neuronal cell death. AD-related mutations in genes coding presenilins are also shown to cause neuronal apoptosis, by directly and indirectly regulating apoptotic signaling cascades. Recent evidence suggests that two intrinsic pathways, mitochondrial dysfunction and endoplasmic reticulum stress, are central in the execution of apoptosis in AD. This review summarizes recent progress of research in this field focused on the molecular mechanisms involved in neuronal apoptosis mediated by organelle dysfunction. PMID- 15750291 TI - Roles of the receptor for advanced glycation endproducts in diabetes-induced vascular injury. AB - Diabetic patients have shorter life span and poorer Quality of Life mainly due to diabetic vascular complications. Recent in vitro and in vivo studies have shown that advanced glycation endproducts (AGE) account for diabetic vascular complications through their engagement of the receptor for AGE (RAGE). In this review, we summarize our recent studies on the roles of the AGE-RAGE system in diabetes-induced vascular injury. In vitro experiments showed that AGE engagement of RAGE leads to changes in endothelial cells (EC) and pericytes, which are characteristic of diabetic microangiopathy. Diabetic RAGE transgenic mice that overexpress RAGE in vascular cells exhibited the exacerbation of the indices of nephropathy and retinopathy, and this was prevented by the inhibition of AGE formation. RAGE overexpression also caused calcium handling impairment in cardiac myocytes. In contrast to the RAGE-overexpressing mice, diabetic RAGE knockout mice showed marked improvement of nephropathy. We found that human vascular cells express a novel splice variant coding for a soluble RAGE protein and named it endogenous secretory RAGE (esRAGE). The esRAGE neutralizes AGE actions on EC and is present in human sera. Individual variations in circulating esRAGE could be a determinant for individual differences in susceptibility or resistance to the development of diabetic vascular complications. The AGE-RAGE system should be, therefore, a candidate molecular target for overcoming diabetic vascular complications. PMID- 15750293 TI - Application study of developmental engineering for livestock production. AB - This paper describes several technical improvements in developmental engineering for livestock production, including their practical utility in the field. The artificial production of monozygotic twins via embryo splitting is shown to increase embryo productivity, while embryo sexing capability provides added value without compromising offspring productivity, with both techniques being adequate for practical field applications. It is also shown that: (1) the development of nuclear transfer utilizing oocytes collected from slaughtered ovaries and matured in vitro enables producing a large number of cloned embryos, (2) the intracytoplasmic injection of somatic cell improves the productivity of nuclear transplantation, and (3) the injection of sperm increases the rate of normal oocytes with male and female pronuclei allowing further preimplantation development. Finally, the removal of cytoplasmic lipid droplets from embryos following centrifugation alters an embryo's intrinsic sensitivity to low temperature allowing long-term preservation. Collectively, these techniques have clearly provided improvements in developmental engineering for livestock production. PMID- 15750292 TI - Studies on reproductive endocrinology in non-human primates: application of non invasive methods. AB - A practical method for the quantitative measurement of estrone conjugates (E1C), pregnanediol-3-glucronide (PdG), follicle stimulating hormone (FSH), and monkey chorionic gonadotropin (mCG) in the excreta of non-human primates were described. In the series of studies, results suggest that 1) urinary and fecal steroid metabolites accurately reflected the same ovarian or testicular events as observed in plasma steroid profiles in captive Japanese macaques, time lags associated with fecal measurements were one day after appearance in urine; 2) these noninvasive methods were applicable to wild and free-ranging macaques for determining reproductive status; 3) hormonal changes during menstrual cycles and pregnancy could be analyzed by measurement of FSH, CG and steroid metabolites in the excreta in captive great apes and macaques; and 4) hormone-behavior relationships of macaques in their natural habitats and social setting could be analyzed. In macaques, between maternal rejection and excreted estrogen, but not excreted progesterone were associated, moreover, in male study, significantly higher levels of fecal cortisol were observed in high-ranking males. In addition, reliable noninstrumented enzyme-linked immunosorbent assay (NELISA) for detection of early pregnancy in macaques was established. These results suggest that the noninvasive characteristic of excreted hormone monitoring provide a stress-free approach to the accurate evaluation of reproductive status in primates. PMID- 15750294 TI - The roles of vitamin A for cytoplasmic maturation of bovine oocytes. AB - Vitamin A is one of the micronutrients which have been implicated in cattle reproduction. In cattle, ingested vitamin A, mainly as beta-carotene (BC) from forages and retinol ester from formula feed, is metabolized and transported to the oocytes and cumulus-granulosa cells in ovarian follicles through binding to various interacting molecules. The active form of vitamin A, retinoic acid (RA), functions as a regulator of gene expression in these targets. Early research showed the positive effects of vitamin A supplementation on bovine fertility in artificial insemination, and several studies on effects of vitamin A metabolites used in other artificial reproductive techniques (ART), including superovulation, ovum pick up, and in vitro maturation culture have provided evidence for the specific roles of vitamin A in oocyte cytoplasmic maturation (acquisition of developmental competence of oocytes during their meiotic maturation period for the embryonic development after fertilization). BC may enhance cytoplasmic maturation by its antioxidant properties which cannot be replaced by RA. Furthermore, RA may promote cytoplasmic maturation of bovine oocytes via its modulatory effects on the gene expression of gonadotrophin receptors, midkine, cyclooxygenase-2, and nitric oxide synthase in cumulus-granulosa cells. PMID- 15750295 TI - First messenger regulation of mammalian sperm function via adenylyl cyclase/cAMP. AB - When released into an appropriate environment, mammalian spermatozoa begin to capacitate and then continue until fully capacitated and able to fertilize. During capacitation in vitro, some cells 'over-capacitate' and undergo spontaneous acrosome reactions; this would be highly undesirable in vivo since already acrosome-reacted spermatozoa are non-fertilizing. Recent studies have revealed that seminal plasma contains several small molecules that bind to specific receptors on the sperm plasma membrane and act as 'first messengers', causing biologically important changes in availability of the 'second messenger' cAMP. Fertilization promoting peptide (FPP), calcitonin and adenosine all regulate cAMP production, stimulating it in uncapacitated spermatozoa and then inhibiting it in capacitated cells; in contrast, angiotensin II stimulates cAMP throughout capacitation. The molecules that regulate cAMP appear to do so via G protein-modulated changes in membrane associated adenylyl cyclases (mACs). Both mouse and human spermatozoa have been shown to have Galphas and Galphai2, as well as several isoforms of mAC, located in the same regions as the specific receptors. Thus spermatozoa possess the required elements for several separate signal transduction pathways, many of which regulate mAC/cAMP and so maintain sperm fertilizing ability. In vivo, such responses could increase the chances of successful fertilization. PMID- 15750296 TI - In vitro attachment of bovine hatched blastocysts on fibronectin is mediated by integrin in a RGD dependent manner. AB - We investigated the effect of extracellular matrix protein on in vitro attachment and outgrowth of bovine hatched blastocysts. In vitro produced bovine hatched blastocysts were cultured on a fibronectin- or laminin-coated Petri dishes. Hatched blastocysts adhered and outgrew on the fibronectin-coated dish whereas no attachment was observed on the laminin-coated dish. The attachment and outgrowth on fibronectin were significantly inhibited in the presence of synthetic peptides containing the Arg-Gly-Asp (RGD) sequence, which interacts with the fibronectin receptor (integrin alpha5beta1), but were not inhibited by the control peptides containing the Arg-Gly-Glu (RGE) sequence. Addition of anti-fibronectin receptor (integrin alpha5beta1) antibody to the culture medium also inhibited the attachment and outgrowth on fibronectin-coated Petri dishes. Subsequently we examined mRNA expression and protein expression of alpha5 and beta1 integrin subunit in the hatched blastocyst by reverse transcription-polymerase chain reaction (RT-PCR) and immunostaining, respectively. Expression of both mRNA and protein were detected in blastocysts. These results indicate that trophectoderm cells of bovine hatched blastocysts have already acquired the ability to adhere and outgrow on fibronectin in vitro by an integrin- mediated manner. PMID- 15750297 TI - Long term consequences of low birthweight on postnatal growth, adiposity and brain weight at maturity in sheep. AB - Low birth weight (LBW) as a result of restricted fetal growth increases the risk for later metabolic diseases and adiposity. However the relationship between LBW and postnatal growth and adult body composition has not been fully investigated. We have used sheep to determine the effects of LBW on postnatal growth and body composition at maturity. LBW was induced by twinning and placental embolization. At birth, LBW lambs were 38% lighter than controls (2.8 +/- 0.2 vs 4.4 +/- 0.3 kg, P<0.05), but had caught up in bodyweight by 8 weeks after birth. At approximately 2.3 years, bodyweights were not different between groups, but there were reductions in absolute (-8%) and relative (-17%) brain weights of LBW sheep (P<0.05) compared to controls. X-ray absorptiometry showed that the mature LBW sheep, compared to controls, had greater amounts of lean muscle (38.1 +/- 1.3 vs 35.3 +/- 0.5 kg, P<0.05) and tended to have more body fat (12.2 +/- 1.2 vs 9.6 +/ 0.9 kg; P=0.1); at autopsy abdominal fat mass was greater in LBW sheep (3.06 +/- 0.26 vs 2.20 +/- 0.25 kg, P<0.05). Plasma leptin concentrations were not different between groups. We conclude that, in sheep, LBW is associated with early postnatal catch-up in body weight, but body composition is permanently altered such that, relative to controls, adiposity is increased and brain weight is decreased. PMID- 15750298 TI - Attempt at in vitro maturation of minke whale (Balaenoptera Bonaerensis) oocytes using a portable CO2 incubator. AB - The present study was conducted to investigate whether a portable CO2 incubator was effective for in vitro maturation (IVM) of bovine, porcine and minke whale oocytes, and the effect of maturation media supplemented with different hormones; porcine follicle stimulating hormone (pFSH), estradiol-17beta (E2), or pregnant mare's serum gonadotropin (PMSG): human chorionic gonadotropin (hCG) for minke whale immature oocytes was also examined. In vitro maturation rates of bovine and porcine oocytes cultured in the portable CO2 incubator were not significantly different from the standard CO2 incubator. In minke whale IVM culture using the portable incubator, the maximum expansion of cumulus mass was observed by pFSH/E2 and PMSG/hCG at the end of IVM culture. Moreover, the IVM culture period was shortened to 28-30 h from 96-120 h previously reported. The proportion of matured oocytes cultured in the medium supplemented with pFSH/E2 (26.7%) was significantly higher (P<0.05) than that with PMSG/hCG (6.9%). The present study indicates that a portable CO2 incubator is a useful device for minke whale IVM culture on a research base ship, and the addition of pFSH/E2 into an IVM medium enhanced cumulus expansion and the proportion of minke whale matured oocytes. PMID- 15750299 TI - Effects of long-term grafting on follicular growth in porcine ovarian cortical grafts xenoplanted to severe combined immunodeficient (SCID) mice. AB - To establish a tool for the study of follicular growth and development, we xenotransplanted small pieces (approximately 1 mm3) of porcine ovarian cortical tissues containing only primordial follicles and small preantral follicles under the capsules of kidneys of severe combined immunodeficient (SCID) mice (8-10 weeks old). The changes in cell proliferation and cell death/apoptosis, and vascularization in xenotransplanted follicles during follicular growth and development were analyzed histochemically at 1-26 weeks after operation. Follicles in grafted ovarian tissues grew rapidly forming an antral cavity (a hallmark of tertiary follicles) at 1 week after grafting. The diameter of the follicles in transplanted tissues ranged from 0.5 to 1.5 mm, from 0.5 to 2.0 mm and from 0.5 to 3.0 mm at 1, 2 and 26 weeks after the operation, respectively. Histological observation of ovarian tissues at 26 weeks after grafting revealed that all grafts had abundant capillary vessels, which invaded from murine organs and surrounded the growing follicles. Grafted small preantral follicles developed to the antral stages at 1 week after grafting and growing antral follicles survived at 26 weeks after grafting. The oocytes in the growing follicles were easily recovered for evaluating the quality. Our simple xenografting system is easy to use and a good experimental tool for the study of folliclular growth and development in porcine ovaries. PMID- 15750300 TI - Effects of ovary storage time and temperature on DNA fragmentation and development of porcine oocytes. AB - The present study was conducted to investigate the effects of storage time and temperature of porcine ovaries on the quality and nuclear maturation in vitro of oocytes obtained from stored ovaries and their subsequent development after in vitro fertilization. The ovaries were stored in physiological saline for 0, 3, 6, 9 and 12 h at various temperatures (4, 15, 25 and 35 C). The pH of follicular fluid obtained from the ovaries, DNA fragmentation of the oocyte nucleus and meiotic competence of oocytes were examined. Some oocytes from ovaries stored at 15, 25 and 35 C for 6 h were fertilized in vitro, and then cultured for 7 days to examine the ability of embryos to develop to the blastocyst stage. When the ovaries were stored at 35 C, the pH of follicular fluid decreased and the proportions of oocytes with DNA fragmented nuclei increased as the storage time was prolonged, and the storage of ovaries for 6, 9 and 12 h resulted in lower maturation rates of oocytes. When the ovaries were stored at 4, 15, 25 and 35 C for 6 h, the storage at higher temperatures (> or =15 C) decreased the pH of follicular fluid and induced nucleic DNA fragmentation in higher proportions of oocytes. None of the oocytes from ovaries stored at 4 C reached metaphase II. The storage of ovaries at 15 C reduced the rates of in vitro fertilized oocytes and subsequent embryo development, but there were no significant differences in the rates of fertilization and blastocyst formation between oocytes from ovaries stored at 25 C and 35 C. Our findings indicate that the storage of ovaries at 25 35 C for 6 h is effective for maintaining the developmental competence of porcine oocytes even though the development rates were lower than those of ovaries stored at 35 C for 3 h. PMID- 15750301 TI - Response of plasma cortisol and progesterone after ACTH challenge in ovariectomized lactating dairy cows. AB - Shortened and weakened estrous expressions could be one of the causes of poor heat detection rate. Non-specific acute stresses are assumed to depress expression of estrus by an increase of plasma progesterone which may originate from the adrenal cortex. The objective of the present study was to examine whether the adrenal cortex can secrete significant amounts of progesterone in response to exogenous adrenocorticotropic hormone (ACTH) in lactating cows. Four cows had estrus synchronized and were ovariectomized in the luteal phase. The cows were given 25 IU ACTH through an indwelling catheter 5 h after catheterization. Blood samples were collected at an interval of 30 min. In 3 of the 4 cows, plasma progesterone concentrations increased significantly 0.5-1.5 h after the first ACTH challenge with a mean peak value of 4.2 +/- 0.4 (S.D.) ng/ml. A similar response was also observed after the second ACTH challenge. Peak plasma progesterone concentrations in the 3 cows after first ACTH challenge were comparable with the progesterone values in the luteal phase of each cow. The results suggest that lactating cows have the capability to secrete a significant amount of progesterone from the adrenal cortex. PMID- 15750302 TI - Ovarian activity and oocyte development during follicular development in pigs at different reproductive phases estimated by the repeated endoscopic method. AB - The aim of the present study was to assess follicular and oocyte development in the same gilts during three phases of their reproductive life [prepuberal gilts (PP; 6.0 months of age), puberal gilts (P; 9.5 months of age) and primiparous sows (S)]. Follicular development was stimulated by the injection of 1,000 IU of equine chorionic gonadotropin (eCG) followed by 500 IU of human chorionic gonadotropin (hCG) 72 h later. Cumulus-oocyte-complexes (COCs) were recovered by endoscopic ovum pick up/aspiration from preovulatory follicles of the left ovary, and the follicular fluid (FF) from the right ovary was collected 34 h after the hCG treatment by endoscopy. Altogether, 19 pigs were used in the PP and P trials and 12 in the S trial. From the left ovaries, 168, 190 and 82 follicles were aspirated and 106, 125 and 42 COCs, respectively, were recovered (recovery rate 61 +/- 27, 63 +/- 21 and 53 +/- 22%, respectively). The mean number of follicles was greater in the P phase than in the PP phase (19.7 +/- 6.8 vs. 15.7 +/- 6.8; p=0.06) and S phases (14.2 +/- 4.0; p<0.05). More uniform oocytes with an expanded cumulus were aspirated in the P and PP phases than in the S phase (90 and 78 vs. 46%; p<0.05). Furthermore, the meiotic configuration in oocytes (T I/M II stage) differed between the three phases (56 and 62 vs. 0%; p<0.05). Progesterone (P4) levels in FF decreased from 590.0 +/- 333.6 (PP) to 249.1 +/- 72.6 (P) and 161.4 +/- 75.2 ng/ml (S) (p<0.05). Estradiol-17beta (E2) levels differed between PP and P gilts and S sows (9.3 +/- 2.9, 21.9 +/- 10.6 and 94.0 +/- 15.9 pg/ml, respectively; p<0.05), and the P4/E2 ratio was 72, 15 and 5, respectively. These results indicate differences in follicular and oocyte development between the reproductive phases investigated. Puberal gilts should preferably be used in IVF and breeding programs. The lower reproductive potential of primiparous sows must be taken into consideration in breeding. Any prediction of lifetime performance based on individual ovarian reactions of prepuberal gilts is unreliable. PMID- 15750303 TI - Transcriptional activity of the 5' upstream region of the porcine glycoprotein hormone alpha subunit gene. AB - Gene expression of the porcine glycoprotein hormone alpha subunit (p-alphaGSU) was examined in LbetaT2 cells, which were established from the anterior pituitary lobe of the immortalized transgenic mouse and produce alphaGSU, and in CHO cells cloned from Chinese hamster ovaries. Expression of the reporter gene fused with p alphaGSU gene upstream in LbetaT2 cells showed that the distal regions -540/-240 and -798/-541 are important for the activation of gene expression. In contrast, the transcriptional activity of the distal region of p-alphaGSU gene was repressed in CHO cells. The region -540/-240 contains an adequate enhancer, called pituitary glycoprotein hormone basal element, whereas the region -798/-541 has no distinguished element. Transfection of the expression vector containing cDNA of a pan-pituitary activator, Ptx1, whose putative binding sites are present scatted in the distal region of the p-alphaGSU gene, revealed unexpectedly that this factor significantly suppressed the expression of p-alphaGSU gene in LbetaT2 cells, indicating that Ptx1 is unrelated to the upregulation in the region -798/ 541. Thus, this study demonstrated for the first time that the distal region 798/-541of the p-alphaGSU gene is indispensable for prominent expression of this gene in which an as yet unidentified factor may participate. PMID- 15750304 TI - Evaluation of the quality of porcine somatic cell nuclear transfer embryo by gene transcription profiles. AB - This study aimed to evaluate the quality of porcine somatic cell nuclear transfer (SCNT) embryos by examining its gene transcription patterns. Embryos were produced by SCNT, intracytoplasmic sperm injection (ICSI) or under different conditions, and transcripts of genes for fibroblast growth factor receptor (FGFr) 2IIIc, FGFr72IIIb, X inactive-specific transcript (Xist), interleukin 6 (IL6), IL6 receptor (IL6r) alpha and c-kit ligand, were detected by real-time RT-PCR. The percentages of embryos in which these transcripts were detected were similar in SCNT and ICSI embryos. On the other hand, the transcriptional levels of the FGFr72IIIb and IL6ralpha genes were 0.5 times less and 2 times more, respectively, in SCNT blastocysts than those of ICSI blastocysts (p<0.05). When nuclear transfer was performed before or after activation of oocytes, embryos in the latter case showed significantly lower frequencies of having FGFr72IIIb (74% vs. 90%) and Xist (3% vs. 33%) transcripts compared to the former case embryos (p<0.05). When two lines of nuclear donor cells with different developmental potencies were used, the transcriptional profiles in the reconstructed embryos did not show any significant differences. Our study suggests that expression profiles of FGFr72IIIb, IL6ralpha, and Xist can be used as markers for the diagnosis of the developmental potency of porcine nuclear transfer embryos. PMID- 15750305 TI - Alteration of reproductive hormone levels in pregnant sows induced by repeated ACTH application and its possible influence on pre- and post-natal hormone secretion of piglets. AB - Prenatal stress has been seen as a reason for reproductive failures in pig offspring mostly originated or mediated by changed maternal functions. Experiments were conducted in pregnant gilts (n=32) to characterize effects of elevated maternal glucocorticoids on the secretion of reproductive hormones (LH, progesterone) during the 1st (EXP 1), 2nd (EXP 2) and 3rd (EXP 3) trimester of pregnancy (TP). Transiently elevated cortisol release was repeatedly achieved by application of 100 IU adenocorticotropic hormone (ACTH) (Synacthen Depot) six times every second day beginning either on day 28 (EXP 1), day 49 (EXP 2) or day 75 of pregnancy (EXP 3). Glucocorticoid concentrations were examined in umbilical blood vessels of fetuses which mothers were subjected to ACTH at 2nd and 3rd TP (EXP 4). Furthermore, the pituitary function of newborn piglets of EXP 2 was checked by a LH-RH challenge test. In sows, LH concentrations were at low basal level (0.1-0.2 ng/ml) but with pulsatory release pattern during each TP. The number of LH pulses/6 h (LSM +/- SE) of saline treated Controls increased with ongoing pregnancy and decreased to the 3rd TP (1.3 +/- 0.2 in EXP 1 vs. 2.0 +/- 0.1 in EXP 2 vs. 1.4 +/- 0.1 in EXP 3, p<0.05). After ACTH treatment the number of LH pulses left unchanged in Experiments 1 and 2 (1.3 +/- 0.2 and 1.5 +/- 0.1) and decreased in EXP 3 (0.8 +/- 0.2, p<0.05). Differences (p<0.05) were obtained comparing the LH pulse number of ACTH and saline treated sows at the 2nd and 3rd TP. Moreover, areas under the curve (AUC) of each LH pulse and of LH over baseline were significantly reduced by treatment. Levels of progesterone increased (p<0.05) for 150 to 170 min after each ACTH application both in EXP 1 and EXP 2, but not in EXP 3. The mean progesterone concentration was different between trimesters, and ACTH and Controls (1st TP: 30.0 +/- 0.9 and 24.4 +/- 0.7 ng/ml; 2nd TP: 35.5 +/- 0.9 and 29.1 +/- 1.0 ng/ml; 3rd TP: 13.6 +/- 0.2 and 13.1 +/- 0.1 ng/ml; p<0.05). In fetuses (n=87) recovered 3 h after ACTH or saline (EXP 4), the plasma cortisol concentrations were significantly increased in umbilical vein (93.7 +/- 5.5 vs. 47.0 +/- 5.3 nmol/l) and artery (95.7 +/- 5.4 vs. 66.4 +/- 5.4 nmol/l), and in periphery (46.8 +/- 5.3 vs. 27.1 +/- 5.3 nmol/l) compared to controls. Plasma ACTH concentrations, however, did not differ in fetuses of both treatment groups. Postnatal LH-RH challenge tests (1st and 28th day post partum) induced LH surges in female piglets (n=67) both of ACTH and saline treated sows, but did not differ between groups (1st day: 7.2 +/- 0.8 vs. 8.1 +/- 0.7 ng/ml; 28th day: 10.5 +/- 1.7 vs. 13.6 +/- 2.2 ng/ml). However, basal LH of piglets whose mothers were submitted to ACTH during 2nd TP was lower on 1st day (1.7 +/- 0.2 vs. 2.3 +/- 0.2 ng/ml, p<0.05) but not on 28th day (1.0 +/- 0.2 vs. 1.1 +/- 0.2 ng/ml). However in both groups, the basal LH was always higher on 1st as on 28th day (p<0.05). Thus, chronic intermittent ACTH administration is able to influence the release pattern of maternal reproductive hormones. However, these findings demonstrate that these effects are dependent on the stage of pregnancy. Furthermore, it was shown that maternal cortisol can cross the placenta during gestation and thus may affect maternal-fetal interactions and, as a result, reproductive function of offspring. PMID- 15750306 TI - Cdk2 activity is essential for the first to second meiosis transition in porcine oocytes. AB - The meiotic progression of Xenopus oocytes has been suggested to depend on the activity of cyclin-dependent kinase 2 (Cdk2). We examined whether Cdk2 is involved in the regulation of mammalian oocyte meiosis by injecting porcine oocytes with anti-Cdk2 antibody. At first, the cross-reactivity of the anti-Cdk2 antibody with Cdc2 kinase was evaluated by immunoprecipitation and immunoblotting experiments using porcine granulosa cell extract, and no cross-reactivity with Cdc2 kinase was observed in the antibody used. In the anti-Cdk2 antibody-injected group, 50.7% of the oocytes were arrested in the second metaphase after 50 h of culture and this rate was significantly lower than those in the non-injected intact oocytes or the oocytes injected with mouse IgG (84.5% and 86.7%, respectively). Most of the other oocytes in the antibody-injected group formed a pronucleus without polar bodies or with only one polar body. The cyclin B1 amount in the antibody-injected and activated oocytes was dramatically decreased compared with that in the intact or mouse IgG-injected oocytes after 50 h of culture. These results suggest that Cdk2 is involved in the meiotic maturation of mammalian oocytes, and that the block of Cdk2 activity results in the failure of cyclin B1 accumulation and second meiosis induction. PMID- 15750307 TI - Effects of partial removal of cytoplasmic lipid on survival of vitrified germinal vesicle stage pig oocytes. AB - This study was designed to investigate whether the partial removal of cytoplasmic lipid from immature pig oocytes prior to vitrification had any positive effects on subsequent maturation, fertilization and early development. Oocytes at the germinal vesicle stage were partially freed from cumulus cells and centrifuged, and then polarized cytoplasmic lipid was removed by micromanipulation. When cultured for 44-48 h, significantly fewer of the centrifuged oocytes reached metaphase II (M-II) than did the non-centrifuged oocytes (approximately 53% vs approximately 68%, respectively); however, no further reduction in the M-II rate was observed when centrifuged oocytes were then delipated prior to culture (approximately 47%). To evaluate their sensitivity to the equilibration and vitrification solutions containing ethylene glycol, non-centrifuged, centrifuged, and delipated oocytes were cultured continuously for several minutes in those solutions, then washed and cultured further; no significant differences in the M II rates (approximately 20-27%) were observed among the three treatment groups. When oocytes were vitrified and then warmed, significantly more delipated oocytes reached M-II in culture (approximately 15%) than did the non-delipated oocytes, whether centrifuged or not (approximately 4% in each group). When delipated, vitrified and matured oocytes were microsurgically injected with frozen-thawed spermatozoa, approximately 39% were activated and male pronucleus formation was observed in approximately 40% of activated oocytes; none developed beyond the 4 cell stage. These results show that maturation in vitro of vitrified pig oocytes can be promoted by partial removal of cytoplasmic lipid prior to vitrification and that the vitrified oocytes can be fertilized, although the embryonic development obtained in this study was limited. PMID- 15750308 TI - Acceleration of follicular development by administration of vascular endothelial growth factor in cycling female rats. AB - To address the role of follicular angiogenesis in the determination of ovulatory follicles and the effects of different vascular endothelial growth factor (VEGF) isoforms on follicular angiogenesis and development, mature female rats were treated with an angiogenic inhibitor (TNP-470), and also with VEGF 120 or 164 at different dosages (0.4, 0.8, 4.0 or 8.0 microg/kg body weight) for 3 days during the estrous cycle. Ovarian follicular angiogenesis, the population of large follicles and ovulation were examined. VEGF 120 (0.8 microg/kg) and 164 (8.0 microg/kg) treatments stimulated follicular angiogenesis in the theca interna layer, while TNP-470 treatment showed severe depression of follicular angiogenesis, and completely inhibited ovulation. After administration of VEGF 120 or 164, the number of healthy preovulatory follicles and ovulated oocytes increased significantly, concomitantly with a decrease in the number of atretic preovulatory follicles. The oocytes ovulated had normal fertilizability and developed to term with the same litter size as in the control rats. Our findings suggest that follicular angiogenesis may be a determinant of follicular development during the periovulatory phase, and that VEGF isoforms may play different important roles in regulating follicular angiogenesis. PMID- 15750309 TI - Genetic and antigenic analyses of bovine respiratory syncytial virus detected in Japan. AB - Genetic and antigenic analyses of bovine respiratory syncytial virus were conducted on 12 field strains from Tohoku and Hokuriku districts in Japan during from 2002 to 2004. On the phylogenetic tree of the nucleotide sequences of the glycoprotein region, the examined strains fell in the same cluster as the strain isolated in Nebraska and were classified as the subgroup III. The examined strains were subdivided into 2 lineages (A, B). Isoleucine 200 of the epitope domain was replaced by threonine as a feature of the lineage B strains. The examined strains showed the nucleotide sequence homologies of 88.3-93.3% with the known Japanese strains classified as the subgroup II and of 86.1-96.6% with those in the subgroup III. No significant difference was found on the neutralization index between the examined strain and the 52-163-13 phylogenetically similar to the Japanese vaccine one. The results suggest that the subgroup III strains have existed in Japan and that epidemics of the strains could be protected due to the present vaccination. PMID- 15750310 TI - Fertility of bitches in which estrus was prevented with implantations of chlormadinone acetate for four years. AB - The recurrence of estrus and fertility after removal of a subcutaneous chlormadinone acetate implant (CMA-I) administered to prevent estrus for 4 years, was investigated in 8 female dogs and the results compared with those for 4 untreated female dogs (control group). The sex hormones present during the estrous cycle were also investigated. There were no significant differences in the estrous cycle after removal of the implant between the CMA-I-treated group and the control group. However, although conception was achieved after mating and no uterine diseases developed in the control group, only 5 (4 dogs, 41.7%) of the 12 cases (6 dogs) in which mating took place at the second to fourth estrus after the removal of CMA-I resulted in pregnancy in the CMA-I-treated group. Furthermore, 6 (75.0%) of the 8 dogs in the CMA-I-treated group developed uterine diseases including pyometra or hydrometra. There were no significant differences in plasma progesterone, LH and prolactin levels between the non-pregnant and pregnant dogs in the CMA-I-treated group or control group. These results suggest that long-term implantation of CMA-I affects fertility after the implant is removed. PMID- 15750311 TI - Expression of a gene encoding Trypanosoma congolense putative Abc1 family protein is developmentally regulated. AB - During the attempt to seek T. congolense species-specific diagnostic antigens, we discovered one cDNA clone (P74) encoding 74 kDa putative abc1 protein (p74) from T. congolense PCF cDNA library. It has been suggested that members of the abc1 family are novel chaperonins and essential for both the mitochondrial electron transfer in the bc 1 complex and the coenzyme Q biosynthesis. Although abc1 protein in yeast has a nuclear or mitochondrial subcellular location, neither nuclear localization signal nor mitochondrial targeting signal was found within p74. Northern blot analysis revealed that the transcription level of P74 mRNA in bloodstream form (BSF) cells were 4 times higher than that in procyclic form cells. Western blot analysis also indicated that p74 was only expressed in T. congolense BSF cells, and revealed that molecular mass of native p74 was not 74 kDa but 56 kDa. This indicates extensive post-translational modification in p74. Although further characterization of p74 will be required, our findings provide implications for CoQ biosynthesis pathway in T. congolense. PMID- 15750312 TI - The relationship between the anticoccidial effects of clindamycin and the development of immunity in the Eimeria pragensis/mouse model of large intestinal coccidiosis. AB - The therapeutic effect of clindamycin on Eimeria pragensis (E. pragensis) infection in C57BL/6 mice was demonstrated by suppression of oocyst production and the appearance of degenerated endogenous stages of parasite in the intestine. Short-term clindamycin treatment, from 1 to 4 days or 4 to 8 days post infection (pi) at a dose of 800 mg/kg/day was effective to reduce clinical symptoms, oocyst production and schizogonic development. Interestingly, the short-term treatment schedules allowed the development of a measurable degree of protective immunity to challenge infection in the treated mice. In contrast, clindamycin treatment for the full 12 days period, which almost completely inhibited clinical symptoms and oocyst output, prevented the full development of protective immunity in the treated mice. All these data indicate that clindamycin is efficacious as an anti eimerian agent and that both early and late endogenous developmental stages of E. pragensis exert a deep influence on the development of effective immunity to challenge infection. PMID- 15750313 TI - Abnormal development of nephrons in claudin-16-defective Japanese black cattle. AB - The kidneys of 37 Japanese Black calves aged 2 to 65 months diagnosed with Claudin 16 (CL-16) defect by the DNA-based test were examined pathologically. The animals exhibited clinical symptoms such as growth impairment, renal failure, overgrowth of hooves, and anemia at a young age. There was no correlation between the time of onset and age. Kidney weights relative to body weight were similar to those in normal animals, but both kidney net weights and size were reduced due to atrophy in animals that showed severe renal dysfunction. Histopathological examination of the kidneys showed reduction in the number of glomeruli, compensatory hypertrophy of glomeruli and tubules, and glomerular and tubular atrophy accompanied by interstitial fibrosis and lymphocytic infiltration. Glomeruli were clearly less in number in the kidneys of CL-16-defective animals than those of normal animals even in the cases with mild lesions. A small number of immature glomeruli and tubules were also detected, suggesting that there were fewer nephrons developed at birth in CL-16-defective animals. It was suggested that a defect of the CL-16 gene is involved in the "abnormal development of nephrons". Immunohistopathological examination of the kidneys showed that the epithelium of thick ascending limb of Henle was stained with anti-CL-16 antibody in the control animals, but not in the affected animals, suggesting a defect of CL-16 in the epithelium of renal tubules in the affected animals. PMID- 15750314 TI - Magnetic resonance imaging of a canine eye with melanoma. AB - An 8-year-old Beagle dog had exophthalmos of the left eye for the last two past months. On ophthalmoscopy, the intraocular lesion could not be evaluated due to the opacity of the cornea. Ultrasonography revealed that the eyeball was distorted in shape and shifted in position, however, the precise lesion could not be identified. On magnetic resonance (MR) imaging, the lesion was observed as hyperintense on T1-weighted and hypointense on T2-weighted images, similar to those reported in human melanoma. The lesion was histologically diagnosed to be malignant intraocular melanoma. Though this is only a case report, canine ocular melanoma may show the similar characteristic MR images as in human uveal malignant melanoma. PMID- 15750315 TI - Effect of nutritional conditions on changes in leukocyte populations in Japanese black calves. AB - To clarify the effect of nutritive conditions on changes in immune cells in Japanese Black (JB) calves during the growth period, leukocyte populations were analyzed in ten healthy JB calves managed in one herd. The calves were divided into two groups: five calves in Group 1 were given insufficient nutrition, and the other five calves in Group 2 received adequate nutrition. The levels of serum total cholesterol and glucose were significantly lower in Group 1 than in Group 2 at 1 month. The numbers of CD3+, CD4+ and CD8+ cells tended to be lower in Group 1 than in Group 2 at months 1 and 2, and the difference in CD4+ was significant at month 2. The number of MHC class-II(+high) cells was significantly lower in Group 1 than in Group 2 at months 1 and 2. These results suggest that adequate nutrition might stimulate an increase in immune cells in calves during the growth period. PMID- 15750316 TI - Detection and isolation of winter dysentery bovine coronavirus circulated in Korea during 2002-2004. AB - Although winter dysentery (WD) has been suspected to occur frequently in Korea, to date the exact epidemiology of WD has remained unknown. Therefore, we investigated the causative agents of WD by using electron microscopy, ELISA, RT PCR, and nested PCR from 97 fecal samples of 32 WD-affected herds collected from 8 provinces during 2002-2004. The bovine coronavirus (BoCV) was consistently detected in all herds with WD. Of other pathogens, only coccidian oocyts were inconsistently but concurrently detected with BoCV. Ten isolates were identified as BoCV by immune electron microscope, immunofluorescent test and ELISA with antiserum to BoCV, and RT-PCR. From these results, it is concluded that WD caused by BoCV occurred in relatively high frequency and was widespread in Korea. The results provide important epidemiological data for the control and establishment of a surveillance system for WD in Korea. PMID- 15750317 TI - The relationship between changes in serum glycine and alanine concentrations in non-essential amino acid and milk production in the transition period in dairy cows. AB - The serum concentration of non-essential amino acid (NEAA) was measured in ten Holstein dairy cows grouped as low production (n=5) and high production (n=5) from one month pre-partum through four months post-partum and the relationship between production and amino acid concentrations was studied. The glycine (Gly)/NEAA ratio and the glycine/alanine ratio of the high production group were significantly higher than the low production group (p<0.01). The observed decrease of the alanine (Ala)/NEAA ratio was more remarkable in the high production group than in the low production group. Measurement of Gly/Ala ratio in serum may be useful for evaluating the nutritional status of peri-parturient dairy cow. PMID- 15750318 TI - Quantification of llama inflammatory cytokine mRNAs by real-time RT-PCR. AB - We have developed a method by which llama cytokine mRNAs can be quantified using real-time reverse transcription polymerase chain reaction (RT-PCR). Total RNA was extracted from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) of llama, reverse transcribed to cDNA, and cytokine profiles for interleukin (IL)-1alpha, IL-1beta, IL-6 and tumor necrosis factor (TNF) alpha were quantified by real-time PCR. The expressions of mRNAs of inflammatory cytokines IL-1alpha, IL-1beta, IL-6 and TNFalpha were upregulated upon stimulation with LPS in a dose- and time-dependent manner. Incubation of PBMCs with 100 and 1,000 pg/ml of LPS for 3 to 6 hr resulted in the acceleration of the mRNA levels of inflammatory cytokines. Here, we describe a highly sensitive and reproducible method to quantify the transcription of llama cytokine mRNAs by real time RT-PCR with the double-stranded DNA-binding dye SYBR Green I. PMID- 15750319 TI - Identification of an cysteine-to-arginine substitution caused by a single nucleotide polymorphism in the canine monoamine oxidase B gene. AB - Monoamine oxidase B catalytically oxidizes biogenic amines such as phenylethylamine and dopamine, and its activity is presumed to be related to particular behavioral traits. In this study, we first identified a single nucleotide polymorphism (T199C) located on the putative third exon of the canine monoamine oxidase B gene, which causes an amino acid substitution from cysteine to arginine. We then examined the allelic frequencies in five dog breeds (Golden Retriever, Labrador Retriever, Maltese, Miniature Schnauzer, and Shiba) and found significant variation among them. The present results suggest that analysis of the monoamine oxidase B polymorphism could be a useful means of elucidating the genetic background of breed-specific behavioral characteristics in dogs. PMID- 15750321 TI - In vitro activity of 24 antimicrobial agents against Staphylococcus and Streptococcus isolated from diseased animals in Japan. AB - A total of 88 Staphylococcus and 61 Streptococcus isolates from diseased animals throughout Japan were examined in 2000 for the minimum inhibitory concentrations of 24 different antimicrobials by the agar dilution method standardized by the Japanese Society of Chemotherapy. The resistance rates to aminobenzylpenicillin (36.4%) and benzylpenicillin (35.2%) were high in Staphylococcus isolates, and those to oxytetracycline (45.9%) and kanamycin (21.3%) were high in Streptococcus isolates. Two isolates resistant to oxacillin harbored the mecA gene. One was Staphylococcus epidermidis derived from a pig with arthritis, and the other Staphylococcus cohnii from a head of cattle with mastitis. PMID- 15750320 TI - Growth characteristics of canine distemper virus in a new cell line CCT cells originated from canine malignant histiocytosis. AB - Canine distemper virus (CDV) growth and the morphological characterization were examined in a cell line established from a canine malignant histiocytosis (CCT cell line). The susceptibility of the CCT cells to 3 CDV strains, FXNO, YSA-TC and MD-77 was shown by detection of the antigen in the indirect fluorescent assay. After passaging 4 and 9 times through the CCT cells, only FXNO strain could produce the syncytia where demonstrated the antigens. Titers of 9 passaged viruses through the CCT cells showed slightly higher in the CCT cells than those in Vero cells. Morphological characterization of karyorrhexis and specific DNA ladder by extracted DNA electrophoresis indicated apoptosis in the CDV infected CCT cells. PMID- 15750322 TI - Increase of antibody titer against Leucocytozoon caulleryi by oral administration of recombinant R7 antigen. AB - In this study, we examined oral administration with recombinant R7 (rR7) antigen expressed in Escherichia coli using chicken leucocytozoonosis subunit vaccine (LV)-vaccinated and unvaccinated chickens. Only LV-vaccinated chickens showed re induction of anti-second-generation schizont (2GS) antibody. Also, LV-vaccinated chickens whose anti-2GS antibody titer was middle-level showed increases of the antibody titer compared to vaccinated-control chickens (P>0.01, >0.05) after oral administration with rR7 antigen. PMID- 15750323 TI - A novel phenomenon predicting the entry into a state of hibernation in Syrian hamsters (Mesocricetus auratus). AB - When Syrian hamsters (Mesocricetus auratus) are bred in a cold and short-day environment, most animals go into hibernation after a certain period of time. However, to date it has not been possible to predict which hamster will enter hibernation. In this study, we subcutaneously implanted thermo-loggers in hamsters bred in the cold environment, and recorded the subcutaneous temperature at short intervals until they went into hibernation. A time series analysis of temperature disclosed that a fall of 0.4 to 0.8 degrees C in subcutaneous temperature was seen 5 to 16 days before entering hibernation, and this phenomenon continued for three days or more. No hamster went into the hibernation without displaying this signal. Although the mechanism by which this phenomenon takes place is not clear, it is a sign from the body, which is useful for indicating if a hamster will enter hibernation shortly. PMID- 15750324 TI - Cloning and expression of goat interleukin-18 gene. AB - We isolated and sequenced a 480 bp cDNA encoding mature goat interleukin-18 (gIL 18) from alveolar macrophages and splenocytes activated with LPS by RT-PCR. The gIL-18 gene was cloned into pET32a (+) vectors and sequenced. Nucleotide sequence of gIL-18 shares high homology with cattle. Fusional expression with pET32a (+) of gIL-18 of about 38kD was obtained by SDS-PAGE analysis after induction by IPTG in the E. Coli BL21 expression system. The recombinant protein can induce IFN gamma production in PBMC. The IL-18 mRNA was constitutively detected in goat alveolar macrophages with or without LPS, While, enhanced expression was detected in splenocytes and liver cells if treated by LPS, and can be weakly detected in Peripheral blood mononuclear cells (PBMCs) treated by activators. Significant deference of IL-18 mRNA level may reflect the capacity to produce mature IL-18 in such tissues. PMID- 15750325 TI - Antibabesial activity of protoberberine alkaloids and 20-hydroxyecdysone from Arcangelisia flava against Babesia gibsoni in culture. AB - Bioassay-guided fractionation of the boiled extract from the stems of Arcangelisia flava led to the isolation of palmatine (1), berberine (2), jatrorrhizine (3), dihydroberberine (4) and 20-hydroxyecdysone (5). The chemical structures of these compounds were elucidated on the basis of their chemical and spectral evidence. The isolated compounds were evaluated for their growth inhibiting effects on Babesia gibsoni in culture for a week. Compounds (1-4) showed significant inhibitions at concentrations from 100 to 1.0 microg/ml, while compound 5 at a concentration of 100 microg/ml, only. PMID- 15750326 TI - Aquisition, mobilization and utilization of cellular iron and heme: endless findings and growing evidence of tight regulation. AB - Iron is fastidiously utilized by living cells, since it is an essential element, but is toxic in excess. Cells take up iron via a transferrin-transferrin receptor dependent endocytotic process. The iron thus taken up is used for essential biological functions including oxygen transport, electron transfer, and DNA synthesis. The intracellular level of iron is tightly controlled, through regulation of the cellular uptake of iron and the sequestering of low molecular labile iron into the storage protein ferritin. The known proteins of iron transport and storage, transferrin, transferrin receptor and ferritin, have been recently linked with a number of newly identified proteins that are responsible for inherited diseases of iron metabolisms and play critical roles in the maintenance of iron homeostasis. These proteins are involved in regulation of intracellular levels of iron, iron transport, and heme transport and the oxygen dependent regulation of gene expression. On the other hand, most iron is transported into mitochondria and immediately used for the biosynthesis of heme in erythroid cells. The heme biosynthesis in mitochondria is coupled with the supply of iron, and the heme, exported from mitochondria, is utilized as prosthetic groups of hemeproteins. Furthermore, non-erythroid and erythroid cells possess the different regulatory systems for the biosynthesis of heme; iron positively regulates the biosynthesis in erythroid cells while heme negatively regulates it in non-erythroid cells. Because of the toxicity and insolubility of heme, the intracellular level of uncommitted heme is maintained at a low concentration (< 10(-9)M). The influx and efflux of heme also help to prevent cytotoxicity. Finally, heme-binding transcriptional factors such as Bach1 and NPAS2 regulate the transcription of several genes involved in the synthesis and degradation of heme-hemeproteins. The discovery of new molecules related to disorders of iron and heme metabolism is ascribable to a complete mechanistic understanding of the cellular network of iron homeostasis. The network of interactions that link iron and heme metabolisms with functions of cellular regulation involving oxidative stress and inflammations contributes to new insights into clinical aspects of disorders. PMID- 15750327 TI - Correction of the maxillary occlusal plane relieves persistent headache and shoulder stiffness. AB - It has been known for many years that deformations of the occlusal plane of the teeth cause indefinite symptoms such as headaches or stiffness of the shoulders. However, how the occlusal plane of the teeth should be corrected remains uncertain. The purpose of this study was to examine whether a correction of the deviation of the maxillary occlusal plane (MOP) from the center of dens of axis vertebrae (DAV) improves symptoms in patients having intractable headache or shoulder-stiffness. Forty patients who complained of dental abnormalities and persistent headache or shoulder-stiffness that had not responded to conventional medical treatment and 17 healthy controls were recruited. All subjects received a lateral cephalometric x-ray examination to measure a distance from the MOP and the center of DAV. In the healthy subjects, both the upper and the lower shift of the MOP from the center of DAV were minimal (the upper shift was 1 +/- 2 [mean +/ S.D.] mm and the lower shift was 4 +/- 4 mm). By contrast, the patients had a significantly greater deviation of the MOP from the center of DAV. Dental adjustment treatment was performed in fourteen patients who had a substantial deviation of the MOP from the center of DAV. Those patients were asked about their symptoms which were scored using a point system and were compared before and after treatment. An adjustment procedure of the MOP passing through the DAV significantly relieved clinical symptoms in these patients (before 42.5 +/- 34.4 vs after 7.0 +/- 8.2, p < 0.01). Correction of the MOP passing through the near center of DAV might be effective in relieving clinical symptoms associated with dental deformities. PMID- 15750328 TI - Low incidence of vascular complications in patients with diabetes mellitus associated with liver cirrhosis as compared with type 2 diabetes mellitus. AB - We compared clinical features and vascular complications of patients with diabetes mellitus associated with liver cirrhosis versus patients with type 2 diabetes mellitus. Subjects were 19 patients (LC-DM group) in whom diabetes was diagnosed after development of liver cirrhosis. Control consisted of 38 patients with type 2 diabetes (T2DM group) matched for sex, age, duration of diabetes, body mass index, treatment, and degree of glycemic control, which was determined by glycoalbumin. The LC-DM group had significantly more smokers, higher serum insulin levels, more insulin resistance calculated by homeostasis model assessment, lower blood counts (white and red blood cells, hemoglobin, and platelets), and lower serum levels of total cholesterol, triglyceride, low density lipoprotein cholesterol and lipoprotein (Lp)(a) than the T2DM group. The incidence of diabetic retinopathy and cerebrovascular disease was significantly lower in the LC-DM group compared to the T2DM group. Logistic regression analysis indicated that Lp(a) and the diabetes duration were significant predictors for the retinopathy, while Lp(a) was a significant predictor for the cerebrovascular complication. In diabetes associated with liver cirrhosis, the incidence of diabetic retinopathy and cerebrovascular disease is lower than in type 2 diabetes mellitus in this study, probably because of lower levels of serum Lp(a). PMID- 15750329 TI - Vitamin B6-responsive ornithine aminotransferase deficiency with a novel mutation G237D. AB - Ornithine aminotransferase (OAT) deficiency (MIM: 258870) is a rare congenital metabolic disorder characterized by gyrate atrophy of the choroid and retina. Here, we report a 37-year-old male with gyrate atrophy of the choroid and retina who has been treated for 18 years. At the age of 7 years, the patient consulted an ophthalmologist due to progressive loss of vision. A large atrophied area was observed in his retina, and OAT deficiency was suspected. At the age of 19 years, amino acid analysis revealed high serum ornithine levels (1,140 nmol/ml), with the normal range being 40-100 nmol/ml. He was treated with vitamin B(6) 300 mg/day for 6 months, which successfully reduced his serum ornithine levels by 20 30%. For 18 years since, his serum ornithine levels have been maintained with vitamin B(6) medication. There was no further impairment of vision or increase in the atrophied area, as judged by ophthalmoscopic examination. OAT activity was undetected in white blood cells of the patient and was 105% and 45% of normal values in his wife and son, respectively. OAT gene analysis revealed a novel mutation of Gly237Asp in exon 7 (710G > A) in both alleles of the patient, while his son was a heterozygote for the mutation. Notably, this novel mutation is associated with a vitamin B6-responsive phenotype. Therefore, early diagnosis and treatment with vitamin B(6) may prevent loss of vision in some patients with OAT deficiency. PMID- 15750330 TI - The impact of high-density barium use in double contrast radiographic methods for gastric cancer screening in Niigata, Japan. AB - Use of high-density barium (= or > 180 w/v%) has been widely spread in community based mass screening for gastric cancer in Japan. However, the impact on outcomes of the screening has not been fully evaluated. Then, we investigated the advantage of high-density barium use by comparing the outcomes between a period with high-density barium and that with low-density barium (< 180 w/v%) each for 3 consecutive years between 1991 and 2002 at 5 medical examination organizations in Niigata Prefecture, Japan. Multivariate logistic regression analysis adjusted for age, sex and medical organization revealed that the introduction of high-density barium decreases the positive finding rate in initial mass screening x-ray examinations by 12% (odds ratio, 0.88; 95% confidence interval, 0.86 - 0.90) and increase the gastric cancer detection rate by diagnostic examination by 15% (odds ratio, 1.15; 95% confidence interval, 1.01-1.31). We conclude that introduction of high-density barium has improved the efficiency of mass screening for gastric cancer by decreasing unnecessary diagnostic examinations and reducing the total personal and public health costs. Thus, nationwide use of high-density barium is recommended for mass gastric cancer screening in Japan. PMID- 15750331 TI - The prevalence of silent ischemia in Turkish patients with type 2 diabetes mellitus. AB - Cardiovascular disease (CVD) is the major cause of death in patients with type 2 diabetes mellitus. However, the diagnosis of CVD is delayed due to concealment of antecedent symptoms by factors such as autonomic neuropathy. In this study, we aimed to investigate the frequency of silent ischemia by using exercise electrocardiogram (ECG). The present study included 500 Turkish patients with type 2 diabetes (male/female: 222/278), who showed no evidence of CAD and angina pectoris or no sign(s) of ischemic changes in resting ECGs. All patients underwent treadmill exercise test according to Bruce protocol, and 62 cases (12.4%) exhibited abnormal changes. These patients identified by exercise ECG consisted of 28 males (28/222, [12.6%]) and 34 females (34/278, [12.2%]) and were then examined by coronary angiography. CAD was diagnosed in 53 individuals by coronary angiography. The abnormalities of exercise test are associated with the age of the patients or the duration of diabetes (p < 0.05). There is no significant difference in the severity of coronary disease or in the prevalence of silent ischemia between male and female patients. However, among the patients identified by exercise ECG females have higher body mass index than males, suggesting that obesity may represent the risk factor of CAD in women with type 2 diabetes. PMID- 15750332 TI - Reliability and validity of the Craig Handicap Assessment and Reporting Technique for Japanese individuals with spinal cord injury. AB - It is necessary and important to quantify the handicap sustained by individuals with spinal cord injury (SCI); however, few instruments are available to measure the level of the disability. One of the best developed and most often used measures is the Craig Handicap Assessment and Reporting Technique (CHART). Nevertheless, a Japanese version has not yet been developed and tested for SCI individuals. The purpose of this study was to develop a Japanese version of the CHART (CHART-J) and to investigate its test-retest reliability and discriminant validity in SCI in Japan. Fifty-four individuals with SCI participated in the test-retest reliability study and 293 participated in the discriminant validity analysis. We have shown that the test-retest reliability coefficients of the CHART-J range from 0.57 to 1.00. In addition, its discriminant validity is acceptable for individuals with SCI in Japan. Therefore, the CHART-J is useful as a measure of disability for Japanese individuals with SCI. PMID- 15750333 TI - Miliary tuberculosis associated with multiple intracranial tuberculomas. AB - In this study we have reported a 12-year old girl patient who visited out-patient clinic with the history of headache and convulsion. The patient was diagnosed as miliary tuberculosis and multiple brain tuberculomas. Miliary infiltration was observed in chest x-ray and high-resolution thorax computed tomography (CT), and multiple tuberculomas surrounded with wide edema was observed in Magnetic Resonance Imaging (MRI). Acid-fast bacilli were detected in inducted sputum and gastric fluid. Focal epileptiform activity was seen in electroencephalography (EEG). The patient was administered antitubercular, anti-edema and antiepileptic therapy. The patient had not experienced convulsion for the second time and EEG had been normal; therefore her treatment was completed within 12 months. Chest x ray and high-resolution thorax CT findings turned to normal and brain MRI findings improved significantly. As a conclusion, tuberculosis disease has very different clinical pattern depending on the organs it involves. The significance of our case is due to the presence of both the miliary tuberculosis and intracranial tuberculomas. The patient was admitted to the hospital due to central nervous system symtoms rather than pulmonary symptoms. PMID- 15750334 TI - Small RNAs: classification, biogenesis, and function. AB - Eukaryotes produce various types of small RNAs of 19-28 nt in length. With rapidly increasing numbers of small RNAs listed in recent years, we have come to realize how widespread their functions are and how diverse the biogenesis pathways have evolved. At the same time, we are beginning to grasp the common features and rules governing the key steps in small RNA pathways. In this review, I will summarize the current classification, biogenesis, action mechanism and function of these fascinating molecules. PMID- 15750335 TI - Expression of gamma-tocopherol methyltransferase transgene improves tocopherol composition in lettuce (Latuca sativa L.). AB - A cDNA encoding gamma-tocopherol methyltransferase (gamma-TMT) from Arabidopsis thaliana was overexpressed in lettuce (Latuca sativa L.) to improve the tocopherol composition. Seven lines of lettuce (T0) containing the gamma-TMT transgene were produced by Agrobacterium-mediated transformation. The inheritance and expression of the transgene were confirmed by DNA and RNA gel blot analyses as well as quantification of tocopherols and gamma-TMT activities. The ratio of alpha-/gamma-tocopherol content (TR) varied from 0.6 to 1.2 in non-transformed plants, while the T0 plants had ratios of 0.8 to 320. The ratio ranged from 0.4 to 544 in 41 T1 progenies of the T0 transgenic line gTM3, and the phenotypic segregation indicated monogenic inheritance of the transgene (i.e., 3:1 = dominant:wild-type classes). There was a tight relationship between the TR phenotype and gamma-TMT activity, and enzyme activities were affected by the copy number and transcript levels of the transgene. The TR phenotype was stably expressed in T2 progenies of T1 plants. The results from this study indicated that a stable inheritance and expression of Arabidopsis gamma-TMT transgene in lettuce results in a higher enzyme activity and the conversion of the gamma tocopherol pool to alpha-tocopherol in transgenic lettuce. PMID- 15750336 TI - Proteasome function is inhibited by polyglutamine-expanded ataxin-1, the SCA1 gene product. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal-dominant neurodegenerative disorder caused by expansion of the polyglutamine tract in the SCA1 gene product, ataxin-1. Using d2EGFP, a short-lived enhanced green fluorescent protein, we investigated whether polyglutamine-expanded ataxin-1 affects the function of the proteasome, a cellular multicatalytic protease that degrades most misfolded proteins and regulatory proteins. In Western blot analysis and immunofluorescence experiments, d2EGFP was less degraded in HEK 293T cells transfected with ataxin 1(82Q) than in cells transfected with lacZ or empty vector controls. To test whether the stability of the d2EGFP protein was due to aggregation of ataxin-1, we constructed a plasmid carrying ataxin-1-Delta114, lacking the self-association region (SAR), and examined degradation of the d2EGFP. Both the level of ataxin-1 Delta114 aggregates and the amount of d2EGFP were drastically reduced in cells containing ataxin-1-Delta114. Furthermore, d2EGFP localization experiments showed that polyglutamine-expanded ataxin-1 inhibited the general function of the proteasome activity. Taken together, these results demonstrate that polyglutamine expanded ataxin-1 decreases the activity of the proteasome, implying that a disturbance in the ubiquitin-proteasome pathway is directly involved in the development of spinocerebellar ataxia type1. PMID- 15750337 TI - Comparative characteristics of three human embryonic stem cell lines. AB - Human embryonic stem (hES) cells have unique features including unlimited growth capacity, expression of specific markers, normal karyotypes and an ability to differentiate. Many investigators have tried to use hES cells for cell-based therapy, but there is little information about the properties of available hES cell lines. We compared the characteristics of three hES cell lines. The expression of SSEA-1, -3, -4, and APase, was examined by immunocytochemistry, and Oct-4 expression was analyzed by RT-PCR. Differentiation of the hES cells in vitro and in vivo led to the formation of embryoid bodies (EBs) or teratomas. We examined the expression of tissue-specific markers in the differentiated cells by semiquantitative RT-PCR, and the ability of each hES cell line to proliferate was measured by flow cytometry of DNA content and ELISA. The three hES cell lines were similar in morphology, marker expression, and teratoma formation. However there were significant differences (P < 0.05) between the differentiated cells formed by the different cell lines in levels of expression of tissue-specific markers such as renin, kallikrein, Glut-2, beta- and delta-globin, albumin, and alpha1-antitrypsin (alpha1-AT). The hES cell lines also differed in proliferative activity. Our observations should be useful in basic and clinical hES cell research. PMID- 15750338 TI - RPK118, a PX domain-containing protein, interacts with peroxiredoxin-3 through pseudo-kinase domains. AB - RPK118 is a sphingosine kinase-1-binding protein that has been implicated in sphingosine 1 phosphate-mediated signaling. It contains a PX (phox homology) domain and two pseudo-kinase domains, and co-localizes with sphingosine kinase-1 on early endosomes. In this study we identified a novel RPK118-binding protein, PRDX3 (peroxiredoxin-3), by yeast two-hybrid screening. The interaction between these proteins was confirmed by pull-down assays and co-immunoprecipitation experiments. Deletion studies showed that RPK118 interacted with PRDX3 through its pseudokinase domains, and with early endosomes through its PX domain. Double immunofluorescence experiments demonstrated that PRDX3 co-localized with RPK118 on early endosomes in COS7 cells. PRDX3 is a member of the antioxidant family of proteins synthesized in the cytoplasm and functioning in mitochondria. Our findings indicate that RPK118 is a PRDX3-binding protein that may be involved in transporting PRDX3 from the cytoplasm to its mitochondrial site of function or to other membrane structures via endosome trafficking. PMID- 15750339 TI - Efficient derivation of new human embryonic stem cell lines. AB - Human embryonic stem (hES) cells, unlike most cells derived from adult or fetal human tissues, represent a potentially unlimited source of various cell types for basic clinical research. To meet the increased demand for characterized hES cell lines, we established and characterized nine new lines obtained from frozen thawed pronucleus-stage embryos. In addition, we improved the derivation efficiency from inner cell masses (to 47.4%) and optimized culture conditions for undifferentiated hES cells. After these cell lines had been maintained for over a year in vitro, they were characterized comprehensively for expression of markers of undifferentiated hES cells, karyotype, and in vitro/in vivo differentiation capacity. All of the cell lines were pluripotent, and one cell line was trisomic for chromosome 3. Improved culture techniques for hES cells should make them a good source for diverse applications in regenerative medicine, but further investigation is needed of their basic biology. PMID- 15750340 TI - Identification of a deoxyribonuclease I inhibitor from a phage-peptide library. AB - Deoxyribonuclease I (DNase I) is a divalent cation dependent endonuclease and thought to be a significant barrier to effective gene delivery. The only known DNase I-specific inhibitor is monomeric actin which acts by forming a 1:1 complex with DNase I. Its use, however, is restricted because of tendency to polymerize under certain conditions. We screened two random phage peptide libraries of complexity 10(8) and 10(9) for DNase I binders as candidates for DNase I inhibitors. A number of DNase I-binding peptide sequences were identified. When these peptides were expressed as fusion proteins with Escherichia coli maltose binding protein, they inhibited the actin-DNase I interaction (IC50 = 0.1-0.7 microM) and DNA degradation by DNase I (IC50 = 0.8-8 microM). Plasmid protection activity in the presence of DNase I was also observed with the fusion proteins. These peptides have the potential to be a useful adjuvant for gene therapy using naked DNA. PMID- 15750341 TI - The Src/PLC/PKC/MEK/ERK signaling pathway is involved in aortic smooth muscle cell proliferation induced by glycated LDL. AB - Low density lipoproteins (LDL) play important roles in the pathogenesis of atherosclerosis. Diabetes is associated with accelerated atherosclerosis leading to cardiovascular disease in diabetic patients. Although LDL stimulates the proliferation of arterial smooth muscle cells (SMC), the mechanisms are not fully understood. We examined the effects of native LDL and glycated LDL on the extracellular signal-regulated kinase (ERK) pathway. Addition of native and glycated LDL to rat aorta SMCs (RASMCs) stimulated ERK phosphorylation. ERK phosphorylation was not affected by exposure to the Ca2+ chelator BAPTA-AM but inhibition of protein kinase C (PKC) with GF109203X, inhibition of Src kinase with PP1 (5 microM) and inhibition of phospholipase C (PLC) with U73122/U73343 (5 microM) all reduced ERK phosphorylation in response to glycated LDL. In addition, pretreatment of the RASMCs with a cell-permeable mitogen-activated protein kinase kinase (MEK) inhibitor (PD98059, 5 microM) markedly decreased ERK phosphorylation in response to native and glycated LDL. These findings indicate that ERK phosphorylation in response to glycated LDL involves the activation of PKC, PLC, and MEK, but is independent of intracellular Ca2+. PMID- 15750342 TI - Polymorphism and expression of isoflavone synthase genes from soybean cultivars. AB - Isoflavones are synthesized by isoflavone synthases via the phenylpropanoid pathway in legumes. We have cloned two isoflavone synthase genes, IFS1 and IFS2, from a total of 18 soybean cultivars. The amino acid residues of the proteins that differed between cultivars were dispersed over the entire coding region. However, amino acid sequence variation did not occur in conserved domains such as the ERR triad region, except that one conserved amino acid was changed in the IFS2 protein of the GS12 cultivar (R374G) and the IFS1 proteins of the 99M06 and Soja99s65 cultivars (A109T, F105I). In three cultivars (99M06, 99M116, and Simheukpi), most of amino acid changes were such that the difference between the amino acid sequences of IFS1 and IFS2 was reduced. The expression profiles of three enzymes that convert naringenin to the isoflavone, genistein, chalcone isomerase (CHI), isoflavone synthase (IFS) and flavanone 3-hydroxylase (F3H) were examined. In general, IFS mRNA was more abundant in etiolated seedlings than mature plants whereas the levels of CHI and F3H mRNAs were similar in the two stages. During seed development, IFS was expressed a little later than CHI and F3H but expression of these three genes was barely detectable, if at all, during later seed hardening. In addition, we found that the levels of CHI, F3H, and IFS mRNAs were under circadian control. We also showed that IFS was induced by wounding and by application of methyl jasmonate to etiolated soybean seedlings. PMID- 15750343 TI - Stress-induced decrease of granule cell proliferation in adult rat hippocampus: assessment of granule cell proliferation using high doses of bromodeoxyuridine before and after restraint stress. AB - Stress is known to inhibit granule cell proliferation in the hippocampus. However, recent studies suggest that the commonly used dose of bromodeoxyuridine (BrdU) is insufficient to label all fractions of granule cells. Furthermore, stress-induced changes in BrdU availability may influence the labeling of newly born cells. To investigate whether changes in BrdU availability affect measurements of stress-induced granule cell proliferation, granule cell proliferation was assessed using injection of high doses of BrdU before and after restraint stress lasting 1 h. In addition, to determine whether stress-induced changes in plasma corticosterone levels were influenced by the BrdU, time dependent changes in plasma corticosterone levels over 2 h after BrdU injection were compared with total accumulated plasma corticosterone levels [as determined by areas under the curve (AUC)]. Restraint stress significantly reduced the numbers of BrdU-labeled cells and clusters in the granule cell layer (GCL) of rats that received BrdU after stress, and decreases of similar magnitude were observed when the rats were given BrdU before stress. BrdU injection enhanced the stress-induced plasma corticosterone response, but there was no difference between the mean AUCs of plasma corticosterone levels of animals injected with BrdU before or after stress. These observations suggest that restraint stress decreases granule cell proliferation, and that this may be influenced by the extent and duration of plasma corticosterone increases rather than by changes in the availability of BrdU. PMID- 15750344 TI - Expression of Arabidopsis phytochelatin synthase 2 is too low to complement an AtPCS1-defective Cad1-3 mutant. AB - Phytochelatins play an important role in heavy metal detoxification in plants as well as in other organisms. The Arabidopsis thaliana mutant cad1-3 does not produce detectable levels of phytochelatins in response to cadmium stress. The hypersensitivity of cad1-3 to cadmium stress is attributed to a mutation in the phytochelatin synthase 1 (AtPCS1) gene. However, A. thaliana also contains a functional phytochelatin synthase 2 (AtPCS2). In this study, we investigated why the cad1-3 mutant is hypersensitive to cadmium stress despite the presence of AtPCS2. Northern and Western blot analyses showed that expression of AtPCS2 is weak compared to AtPCS1 in both roots and shoots of transgenic Arabidopsis. The lower level of AtPCS2 expression was confirmed by RT-PCR analysis of wild type Arabidopsis. Moreover, no tissue-specific expression of AtPCS2 was observed. Even when AtPCS2 was under the control of the AtPCS1 promoter or of the cauliflower mosaic virus 35S promoter (CaMV 35S) it was not capable of fully complementing the cad1-3 mutant for cadmium resistance. PMID- 15750345 TI - Transduced Tat-SOD fusion protein protects against ischemic brain injury. AB - Reactive oxygen species (ROS) are implicated in reperfusion injury after transient focal cerebral ischemia. The antioxidant enzyme, Cu,Zn-superoxide dismutase (SOD), is one of the major means by which cells counteract the deleterious effects of ROS after ischemia. Recently, we reported that when Tat SOD fusion protein is transduced into pancreatic beta cells it protects the beta cells from destruction by relieving oxidative stress in ROS-implicated diabetes (Eum et al., 2004). In the present study, we investigated the protective effects of Tat-SOD fusion protein against neuronal cell death and ischemic insults. When Tat-SOD was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Immunohistochemical analysis revealed that Tat-SOD injected intraperitoneally (i.p.) into mice has access to various tissues including brain neurons. When i.p. injected into gerbils, Tat-SOD prevented neuronal cell death in the hippocampus in response to transient fore-brain ischemia. These results suggest that Tat-SOD provides a strategy for therapeutic delivery in various hu-man diseases, including stroke, related to this anti-oxidant enzyme or to ROS. PMID- 15750346 TI - Critical role of the cysteine 323 residue in the catalytic activity of human glutamate dehydrogenase isozymes. AB - The role of residue C323 in catalysis by human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) was examined by substituting Arg, Gly, Leu, Met, or Tyr at C323 by cassette mutagenesis using synthetic human GDH isozyme genes. As a result, the Km of the enzyme for NADH and alpha-ketoglutarate increased up to 1.6-fold and 1.1-fold, respectively. It seems likely that C323 is not responsible for substrate-binding or coenzyme-binding. The efficiency (kcat/Km) of the mutant enzymes was only 11-14% of that of the wild-type isozymes, mainly due to a decrease in kcat values. There was a linear relationship between incorporation of [14C]p-chloromercuribenzoic acid and loss of enzyme activity that extrapolated to a stoichiometry of one mol of [14C] incorporated per mol of monomer for wild type hGDHs. No incorporation of [14C]p-chloromer-curibenzoic acid was observed with the C323 mutants. ADP and GTP had no effect on the binding of p chloromercuribenzoic acid, suggesting that C323 is not directly involved in allosteric regulation. There were no differences between the two hGDH isozymes in sensitivities to mutagenesis at C323. Our results suggest that C323 plays an important role in catalysis by human GDH isozymes. PMID- 15750347 TI - Widespread occurrence of small inversions in the chloroplast genomes of land plants. AB - Large inversions are well characterized in the chloroplast genomes of land plants. In contrast, reports of small inversions are rare and involve limited plant groups. In this study, we report the widespread occurrence of small inversions ranging from 5 to 50 bp in fully and partially sequenced chloroplast genomes of both monocots and dicots. We found that small inversions were much more common than large inversions. The small inversions were scattered over the chloroplast genome including the IR, SSC, and LSC regions. Several small inversions were uncovered in chloroplast genomes even though they shared the same overall gene order. The majority of these small inversions were located within 100 bp downstream of the 3' ends of genes. All had inverted repeat sequences, ranging from 11 to 24 bp, at their ends. Such small inversions form stem-loop hairpin structures that usually have the function of stabilizing the corresponding mRNA molecules. Intra-molecular recombination between the inverted sequences in the stem-forming regions are responsible for generating flip-flop orientations of the loops. The presence of two different orientations of the stem loop in the trnL-F noncoding region of a single species of Jasminum elegans suggests that a short inversion can be generated within a short period of time. Small inversions of non-coding sequences may influence sequence alignment and character interpretation in phylogeny reconstructions, as shown in nine species of Jasminum. Many small inversions may have been generated by parallel or back mutation events during chloroplast genome evolution. Our data indicate that caution is needed when using chloroplast non-coding sequences for phylogenetic analysis. PMID- 15750348 TI - DNA sequence analysis of 1-nitropyrene-4,5-oxide and 1-nitropyrene-9,10-oxide induced mutations in the hprt gene of Chinese hamster ovary cells. AB - Nitropyrene, the predominant nitropolycyclic hydrocarbon found in diesel exhaust, is a mutagenic and tumorigenic environmental pollutant that requires metabolic activation via nitroreduction and ring oxidation. In order to determine the role of ring oxidation in the mutagenicity of 1-nitropyrene, its oxidative metabolites, 1-nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide, were synthesized and their mutation spectra were determined in the coding region of hprt gene of CHO cells by a PCR amplification of reverse-transcribed hprt mRNA, followed by a DNA sequence analysis. A comparison of the two metabolites for mutation frequencies showed that 1-nitropyrene 9,10-oxide was 2-times higher than 1-nitropyrene 4,5-oxide. The mutation spectrum for 1-nitropyrene 4,5-oxide was base substitutions (33/49), one base deletions (11/49) and exon deletions (5/49). In the case of 1-nitropyrene 9,10-oxide, base substitutions (27/50), one base deletions (15/50), and exon deletions (8/50) were observed. Base substitutions were distributed randomly throughout the hprt gene. The majority of the base substitutions in mutant from 1-nitropyrene 4,5-oxide treated cells were A-->G transition (15/33) and G-->A transition (8/33). The predominant base substitution, A-->G transition (11/27) and G-->A transition (8/27), were also observed in mutant from 1-nitropyrene 9,10-oxide treated cells. The mutation at the site of adenine and guanine was consistent with the previous results, where the sites of DNA adduct formed by these compounds were predominant at the sites of purines. A comparison of the mutational patterns between 1-nitropyrene 4,5 oxide and 1-nitropyrene 9,10-oxide showed that there were no significant differences in the overall mutational spectrum. These results indicate that each oxidative metabolite exhibits an equal contribution to the mutagenicity of 1 nitropyrene, and ring oxidation of 1-nitropyrene is an important metabolic pathway to the formation of significant lethal DNA lesions. PMID- 15750349 TI - In vitro characterization of protein kinase CKII beta mutants defective in beta beta dimerization. AB - Protein kinase CKII is composed of two catalytic (alpha or alpha') subunits and two regulatory (beta) subunits. The beta subunit mediates tetramer formation through beta-beta homodimerization and alpha-beta heterodimerization. In a previous study R26 and R75, point mutants of CKIIb defective in beta-beta dimerization, were isolated. In the present work we characterized these CKIIbeta mutants in vitro. Purified R26 and R75 bound to CKIIalpha but were defective in binding to CKIIbeta. R75 stimulated the catalytic activity of CKII whereas R26 gave little stimulation, and poly-L-lysine increased the stimulation of catalytic activity by R26 or R75. Circular dichroism and intrinsic fluorescence data pointed to different conformational changes in R26 and R75. Molecular modeling of these mutants provides an explanation of the difference in their ability to interact with CKIIbeta and to activate CKIIalpha. PMID- 15750350 TI - Up-regulation of glutathione biosynthesis in NIH3T3 cells transformed with the ETV6-NTRK3 gene fusion. AB - The ETV6-NTRK3 gene fusion, first identified in the chromosomal translocation in congenital fibrosarcoma, encodes a chimeric protein tyrosine kinase with potent transforming activity. ETV6-NTRK3-dependent transformation involves the joint action of NTRK3 signaling pathways, and aberrant cell cycle progression resulting from activation of Mek1 and Akt. The level of glutathione (GSH) was found to be markedly increased in ETV6-NTRK3-transformed NIH3T3 cells. The activities of the two GSH biosynthetic enzymes as well as of glutathione peroxidase, together with their mRNAs, were also higher in the transformed cells. The transformed cells were able to grow in the presence of GSH-depleting agents, whereas the control cells were not. L-Buthionine-(S,R)-sulfoximine (BSO) inhibited activation of Mek1 and Akt in the transformed NIH3T3 cells. These observations imply that up regulation of GSH biosynthesis plays a central role in ETV6-NTRK3-induced transformation. PMID- 15750351 TI - A role for the carbohydrate portion of ginsenoside Rg3 in Na+ channel inhibition. AB - We showed recently that ginsenosides inhibit the activity of various types of ion channel. Here we have investigated the role of the carbohydrate component of ginsenoside Rg3 in the inhibition of Na+ channels. The channels were expressed in Xenopus oocytes by injecting cRNAs encoding rat brain Nav1.2 alpha and beta1 subunits, and analyzed by the two-electrode voltage clamp technique. Treatment with Rg3 reversibly inhibited the inward Na+ peak current (INa) with an IC50 of 32.2 +/- 4.5 microM, and the inhibition was voltage-dependent. To examine the role of the sugar moiety, we prepared a straight chain form of the second glucose and a conjugate of this glucose with 3-(4-hydroxyphenyl) propionic acid hydrazide (HPPH). Neither derivative inhibited INa. Treatment with the carbohydrate portion of ginsenoside Rg3, sophorose [beta-D-glucopyranosyl (1-->2)- beta glucopyranoside], or the aglycone (protopanaxadiol), on their own or in combination had no effect on INa. These observations indicate that the carbohydrate portion of ginsenoside Rg3 plays an important role in its effect on the Na+ channel. PMID- 15750352 TI - Combined effect of heptaplatin and ionizing radiation on human squamous carcinoma cell lines. AB - Heptaplatin, cis-malonato [(4R,5R)-4,5-bis (amino-methyl)-2-isopropyl-1,3 dioxolane] platinum(II) (SKI-2053R, Sunpla) is a new platinum derivative with anti-tumor activity comparable to cisplatin on various cancer cell lines. Preclinical studies suggest that it is less nephrotoxic than cisplatin. This study was undertaken to examine the combined effect of heptaplatin and ionizing radiation on two established human squamous carcinoma cell lines (NCI-H520, SQ20B). The cytotoxic activity of heptaplatin was concentration-dependent in both cell lines. When low dose heptaplatin was combined with high dose ionizing radiation, there was an additive cytotoxic effect on NCI-H520 cells (P < 0.05), while a moderate dose of heptaplatin and a low dose of ionizing radiation had an additive cytotoxic effect on the growth of SQ20B cells (P < 0.05). FACS analysis and DAPI staining showed that their additive cytotoxic effects were correlated with the induction of apoptosis. Further studies are warranted using heptaplatin and ionizing radiation in squamous cell carcinoma as a substitute for cisplatin. PMID- 15750355 TI - Homeopathy: a view from the outside. PMID- 15750353 TI - Identification and characterization of a putative cyclic nucleotide-gated channel, CNG-1, in C. elegans. AB - Cyclic nucleotide-gated (CNG) channels encoded by the tax-4 and tax-2 genes are required for chemosensing and thermosensing in the nematode C. elegans. We identified a gene in the C. elegans genome, which we designated cng-1, that is highly homologous to tax-4. Partial CNG-1 protein tagged with green fluorescent protein was expressed in several sensory neurons of the amphid. We created a deletion mutant of cng-1, cng-1 (jh111), to investigate its in vivo function. The mutant worms had no detectable abnormalities in terms of their basic behavior or morphology. Whereas tax-4 and tax-2 mutants failed to respond to water-soluble or volatile chemical attractants, the cng-1 null mutant exhibited normal chemotaxis to such chemicals and a tax-4;cng-1 double mutant had a similar phenotype to tax 4 single mutants. Interestingly, cng-1 and tax-4 had a synergistic effect on brood size. PMID- 15750356 TI - Cosmic connections: holism as a universal trait. PMID- 15750357 TI - The importance of mechanical forces. PMID- 15750358 TI - Ngemm'dtrrr's story. PMID- 15750359 TI - A pilot, randomized, double-blinded, placebo-controlled trial of individualized homeopathy for symptoms of estrogen withdrawal in breast-cancer survivors. AB - OBJECTIVE: To pilot an investigation of individualized homeopathy for symptoms of estrogen withdrawal in breast cancer survivors. DESIGN: Randomized, double blinded, placebo-controlled trial. SETTING: Outpatient department of a National Health Service (NHS) homeopathic hospital. PARTICIPANTS: Fifty-seven (57) women met inclusion criteria and 53 were randomized to the study. INTERVENTION: After 2 weeks of baseline assessment, all participants received a consultation plus either oral homeopathic medicine or placebo, assessed every 4 weeks for 16 weeks. OUTCOME MEASURES: The primary outcome measures were the activity score and profile score of the Measure Yourself Medical Outcome Profile (MYMOP). RESULTS: Eighty-five percent (85%) (45/53) of women completed the study. There was no evidence of a difference seen between groups for either activity (adjusted difference =-0.4, 95% confidence interval CI -1.0 to 0.2, p = 0.17) or profile scores (adjusted difference = -0.4, 95% CI -0.9 to 0.1, p = 0.13) using this trial design, although post hoc power calculations suggests that 65-175 would be needed per group to detect differences of this magnitude with sufficient precision. Clinically relevant improvements in symptoms and mood disturbance were seen for both groups over the study period. CONCLUSION: Improvements were seen for symptom scores over the study period. However, presuming these improvements were caused by the individualized homeopathic approach, the study failed to show clearly that the specific effect of the remedy added further to the nonspecific effects of the consultation. Future trial design must ensure adequate power to account for the nonspecific impact of such complex individualized interventions while pragmatic designs may more readily answer questions of clinical and cost effectiveness. PMID- 15750360 TI - Homeopathy for menopausal symptoms in breast cancer survivors: a preliminary randomized controlled trial. AB - OBJECTIVES: To carry out a preliminary trial evaluating the effectiveness of two types of homeopathy for the treatment of menopausal symptoms in breast cancer survivors. DESIGN: Randomized, double-blinded, placebo-controlled. SETTINGS/LOCATION: Private medical clinic, Seattle, WA. SUBJECTS: Women with a history of breast cancer who had completed all surgery, chemotherapy, and radiation treatment and who had an average of at least three hot flashes per day for the previous month. INTERVENTIONS: Subjects were randomized to receive either an individualized homeopathic single remedy, a homeopathic combination medicine, or placebo. Patients were seen by homeopathic providers every 2 months for 1 year. OUTCOME MEASURES: Hot flash frequency and severity, Kupperman Menopausal Index (KMI), Short Form 36 (SF-36). RESULTS: There was no significant difference found in the primary outcome measure, the hot flash severity score, although there was a positive trend in the single remedy group during the first 3 months of the study (p = 0.1). A statistically significant improvement in general health score in both homeopathy groups (p < 0.05) on the SF-36 after 1 year was found. Evidence of a homeopathic "drug proving" in the subjects receiving the homeopathic combination medicine who were not taking tamoxifen also was found. CONCLUSIONS: Small sample size precludes definitive answers, but results from this preliminary trial suggest that homeopathy may be of value in the treatment of menopausal symptoms and improving quality of life, especially in those women not on tamoxifen. Larger studies should be carried out that also include healthy women who want to avoid hormone replacement therapy. PMID- 15750361 TI - Genomic profiling of neutrophil transcripts in Asian Qigong practitioners: a pilot study in gene regulation by mind-body interaction. AB - BACKGROUND AND OBJECTIVES: The great similarity of the genomes of humans and other species stimulated us to search for genes regulated by elements associated with human uniqueness, such as the mind-body interaction. DNA microarray technology offers the advantage of analyzing thousands of genes simultaneously, with the potential to determine healthy phenotypic changes in gene expression. The aim of this study was to determine the genomic profile and function of neutrophils in Falun Gong (FLG, an ancient Chinese Qigong) practitioners, with healthy subjects as controls. SUBJECTS AND DESIGN: Six (6) Asian FLG practitioners and 6 Asian normal healthy controls were recruited for our study. The practitioners have practiced FLG for at least 1 year (range, 1-5 years). The practice includes daily reading of FLG books and daily practice of exercises lasting 1-2 hours. Selected normal healthy controls did not perform Qigong, yoga, t'ai chi, or any other type of mind-body practice, and had not followed any conventional physical exercise program for at least 1 year. Neutrophils were isolated from fresh blood and assayed for gene expression, using microarrays and RNase protection assay (RPA), as well as for function (phagocytosis) and survival (apoptosis). RESULTS: The changes in gene expression of FLG practitioners in contrast to normal healthy controls were characterized by enhanced immunity, downregulation of cellular metabolism, and alteration of apoptotic genes in favor of a rapid resolution of inflammation. The lifespan of normal neutrophils was prolonged, while the inflammatory neutrophils displayed accelerated cell death in FLG practitioners as determined by enzyme-linked immunosorbent assay. Correlating with enhanced immunity reflected by microarray data, neutrophil phagocytosis was significantly increased in Qigong practitioners. Some of the altered genes observed by microarray were confirmed by RPA. CONCLUSION: Qigong practice may regulate immunity, metabolic rate, and cell death, possibly at the transcriptional level. Our pilot study provides the first evidence that Qigong practice may exert transcriptional regulation at a genomic level. New approaches are needed to study how genes are regulated by elements associated with human uniqueness, such as consciousness, cognition, and spirituality. PMID- 15750362 TI - Qigong for schoolchildren: a pilot study. AB - OBJECTIVE: To evaluate the effects of qigong lessons on schoolchildren in terms of their achievements at school, social behavior, and general health. DESIGN: A controlled intervention pilot study was conducted with children in two second grade classes at an elementary school and in two eighth-grade classes at a high school. SETTING/LOCATION AND INTERVENTION: One class from each school received qigong lessons for 20 minutes at least twice weekly over a period of 6 months, while the control class from the same school received no intervention. Two additional classes at two elementary schools participated in the qualitative analysis only. MEASURES: Teachers, parents, and students answered standardized questionnaires at the beginning of the study and after 6 months, covering complaints, concentration, creativity, grades, quality of life (QOL) (assessed by the KINDL questionnaire), and social behavior. In addition to this quantitative study, we conducted in-depth, semi-structured interviews with all participating teachers at the end of the project to evaluate individual effects. RESULTS: Of a total of 90 participants in the study (52% boys and 48% girls), 40 students attended elementary school (20 per class, mean age 7.4 +/- 0.5 years) and 50 high school (25 per class, mean age 13 +/- 0.8 years). Factor analysis yielded three scales for the teacher questionnaire (learning process, social behavior, appropriate behavior) and four scales for the parent questionnaire (creativity, concentration, well-being, restlessness). The qigong group showed significantly better results in the teacher questionnaire (especially for appropriate behavior) and in grades in comparison to the control group, but no effect was found in the parent questionnaire, medical complaints, sick days, or in the children's assessment of QOL. Qualitative analyses indicated a relevant decrease of individual complaints for some children in the qigong group. CONCLUSIONS: The feasibility of integrating qigong in school lessons was shown. Six months after starting qigong lessons, schoolchildren improved in social behavior and showed stable grades, while inappropriate behavior decreased, compared to the control. Combining quantitative and qualitative research methods appeared to be useful to detect the effects of qigong in individuals. Due to the limitations of our pilot study, further studies with larger sample sizes and long-term follow-up are needed. PMID- 15750363 TI - The use of an herbal formula by hospital care workers during the severe acute respiratory syndrome epidemic in Hong Kong to prevent severe acute respiratory syndrome transmission, relieve influenza-related symptoms, and improve quality of life: a prospective cohort study. AB - OBJECTIVES: To investigate the efficacy of an herbal formula in the prevention of severe acute respiratory syndrome (SARS) transmission among health care workers. The secondary objectives are to investigate quality of life (QOL) and symptomology changes among supplement users, and to evaluate the safety of this formula. DESIGN: Controlled clinical trial. SETTINGS: Hong Kong during epidemic of SARS. SUBJECTS: Two cohorts of health care workers from 11 hospitals in Hong Kong, 1 using an herbal supplement for a 2-week period (n = 1063) and a control cohort comprising all other health care workers who did not receive the supplement (n = 36,111) were compared prospectively. INTERVENTIONS: Taking an herbal supplement for a 2-week period. OUTCOME MEASURES: SARS attack rates and changes in quality of life and influenza-like symptoms were also examined at three timepoints among herbal supplement users. RESULTS: None of the health care workers who used the supplement subsequently contracted SARS compared to 0.4% of the health care workers who did not use the supplement (p = 0.014). Improvements in influenza-like symptoms and quality of life measurements were also observed among herbal supplement users. Less than 2% reported minor adverse events. CONCLUSION: The results of this pilot study suggest that there is a good potential of using Traditional Chinese Medicine (TCM) supplements to prevent the spread of SARS. PMID- 15750364 TI - Effects of imagery, critical thinking, and asthma education on symptoms and mood state in adult asthma patients: a pilot study. AB - OBJECTIVE: To compare biologically targeted imagery (BTI) and critical thinking asthma management (CTAM) outcomes. DESIGN: Participants were randomized to BTI (group 1, G1) or CTAM (group 2, G2) in a 2 x 2 x 4 design (BTI/CTAM x pretest/post-test x weeks [3 week averaged intervals of symptoms and peak flows]). Interventions were asthma education plus treatment (BTI or CTAM for two 2-hour sessions per week for 6 weeks). For BTI, data collection (symptoms, lung function) occurred preintervention (3 week baseline), during the intervention (6 weeks), and postintervention (6 weeks). For CTAM, data collection occurred at wait-list control (WLC) (12 weeks extended baseline), preintervention (3 weeks), during the intervention (6 weeks), and postintervention (6 weeks). SETTING: Alaska Regional Hospital, Anchorage. SUBJECTS: Seventy (70) adults (53 women, 17 men) with asthma. INTERVENTIONS: WLC record keeping, BTI, or CTAM. OUTCOME MEASURES: Dependent variables included asthma symptoms (wheezing, coughing, sleep, activity, attacks, peak flow) and self-report assessments of Profiles of Mood States (POMS-BI) (anxiety, hostility, depression, uncertainty, fatigue, confusion); Knowledge, Attitude, and Self-Efficacy Asthma Questionnaire (KASE AQ); Health Attribution Test (HAT) for locus of control (LoC); and the Revised Asthma Problem Behavior Checklist (RAPBC). RESULTS: Analyses of covariance with repeated measures contrasted BTI pre- to post-tests, time periods, and WLC; CTAM pre- to post-tests, time periods, and extended baseline WLC; and BTI to CTAM outcomes. WLC improved all POMS-BI scores except anxiety, increased internal LoC, and reduced problematic behaviors. Compared to WLC, BTI reduced wheezing, anxiety, and chance LoC, and increased asthma knowledge, attitude, and self efficacy. Compared to CTAM, BTI reduced wheezing and chance LoC, increased internal LoC, and improved 6 POMS-BI scores. Compared to extended baseline WLC, CTAM increased asthma knowledge, attitude, self-efficacy, internal LoC, and peak flow. CONCLUSIONS: Both interventions significantly improved symptoms and asthma management more than record keeping. Contrary to hypothesis, BTI produced better outcomes than CTAM. PMID- 15750365 TI - Interactive Guided Imagery therapy with medical patients: predictors of health outcomes. AB - OBJECTIVES: To assess whether Interactive Guided Imagery (IGI) is helpful to medical patients and to identify factors that contribute to positive outcomes. DESIGN: A prospective cohort study of 323 medical patients who received 6 IGI sessions on a weekly basis. Patients and practitioners completed questionnaires at the beginning, middle, and end of the 6 IGI sessions. The questionnaires assessed the patients' ability to do IGI, the quality of the practitioner-patient interaction, possible confounding variables, and enabling factors. The hypothesis was that measures of the process of doing IGI and the practitioner-patient relationship would predict outcomes. SUBJECTS/SETTING: The subjects were all patients seeking treatment at California Pacific Medical Center in San Francisco, and Marin General Hospital in Greenbrae, CA. INTERVENTION: Using IGI interactively is a cognitive-behavioral intervention designed to help patients relax by using mental images to discover and cultivate healing intentions, and to reflect on the meaning of these images. MEASURES: The individual measures to assess the patients' ability to do IGI and measures of the practitioner-patient relationship were factor-analyzed to use as predictor variables in a multiple regression. Similarly, the questionnaire items measuring cognitive, emotional, behavioral, and spiritual benefits of IGI were factor-analyzed into factors representing "insight" and "all other" benefits. RESULTS: The multiple regression shows that both process and practitioner-patient interaction factors significantly contributed to a combined 40% of the variance in patients' ratings of insight into the nature of their problem and to becoming aware of an aspect of self, F(4,56) = 9.4, p < 0.005. The same process and interaction factors were less strongly related to the other outcomes, r2 = 0.14, F(4,56) = 2.3, p = 0.06. None of the demographic, confounding, or enabling factors was related to the outcome measures. CONCLUSIONS: The process of doing IGI and the relationship with the practitioner were both independently associated with the patients' insight into their health problems. PMID- 15750367 TI - Human biofield and intention detection: individual differences. AB - OBJECTIVE: To evaluate a battery of biofield awareness tasks that address bioelectromagnetic and consciousness related mechanisms of action, and examine individual differences in integrative biofield awareness (IBA). METHODS: Six (6) biofield awareness tasks were designed: 2 involved the experimenter placing his or her hands near the subject, 2 involved intense staring with associated eye movements approximately 3' from the subject, and 2 involved gentle intention with virtually no movement. Each task required a binary response from the subject. There were 10 trials per task for a total of 60 trials; blocks of 6 trials contained one of each task. Subjects were 165 undergraduate psychology students at the University of Arizona Tucson, AZ. Subjects were also assessed on their awareness of their own biofields, and they filled out various questionnaires, including estimates of how well they thought they would do and their openness to spiritual beliefs and experiences. RESULTS: Percent IBA accuracy for the entire sample (n = 165) was 57.7 +/- 10.3% and was significantly above chance (50%) performance (t = 9.58, p < 0.0000001). Each of the 6 tasks was individually significant. Subjects significantly (t = -2.72; p < 0.007) underestimated their IBA performance below chance (mean, 46.1 +/- 18.4%). However, higher estimates predicted higher IBA (r = 0.26, n = 164, p < 0.0008). Measures of subjects' self awareness of their own biofields, as well as belief in, and experience of, extrasensory perception (ESP) also predicted higher IBA. CONCLUSIONS: The findings support claims of energy healers that biofield awareness can be modulated both bioelectromagnetically (locally) and via conscious intent (distally), and that individual differences in biofield awareness are related to self-awareness and sensitivity to others. PMID- 15750366 TI - Gut feelings, intuition, and emotions: an exploratory study. AB - OBJECTIVE: Investigate whether the gut feelings of one person, as measured with an electrogastrogram (EGG), respond to the emotions of a distant person. DESIGN: In a double blind protocol, EGG activity was recorded in an individual relaxing in a heavily shielded chamber while, at a distance, a second person periodically viewed the live video image of the first person along with stimuli designed to evoke positive, negative, calming, or neutral emotions. SUBJECTS: Twenty-six (26) pairs of healthy adult volunteers. OUTCOME MEASURES: EGG maximum values recorded while the distant person was exposed to emotional stimuli were compared to similar values recorded during exposure to neutral stimuli. RESULTS: EGG maximums were significantly larger on average when the distant person was experiencing positive (p = 0.006) and negative (p = 0.0009) emotions, as compared to neutral emotions. Nonparametric bootstrap procedures were employed to evaluate these differences, and the results survive correction for multiple analyses. CONCLUSIONS: EGG activity increases in response to the emotions of a distant person, beyond the influence of ordinary sensory interactions. Relationships commonly reported between gut feelings and intuitive hunches may share a common, poorly understood, perceptive origin. PMID- 15750368 TI - Cranial manipulation can alter sleep latency and sympathetic nerve activity in humans: a pilot study. AB - OBJECTIVE: To determine if cranial manipulation is associated with altered sleep latency. Furthermore, we investigated the effects of cranial manipulation on muscle sympathetic nerve activity (MSNA) as a potential mechanism for altered sleep latency. DESIGN: Randomized block design with repeated measures. SETTING: The Integrative Physiology and Manipulative Medicine Departments, University of North Texas Health Science Center, Fort Worth, TX. SUBJECTS: Twenty (20) healthy volunteers (12 male, 8 female; age range, 22-35 years) participated in this investigation. INTERVENTIONS: Subjects were exposed to 3 randomly ordered treatments: compression of the fourth ventricle (CV4), CV4 sham (simple touch), and control (no treatment). OUTCOME MEASURES: Sleep latency was assessed during each of the treatments in 11 subjects, using the standard Multiple Sleep Latency Test protocol. Conversely, directly recorded efferent MSNA was measured during each of the treatments in the remaining 9 subjects, using standard microneurographic technique. RESULTS: Sleep latency during the CV4 trial was decreased when compared to both the CV4 sham or control trials (p < 0.05). MSNA during the CV4-induced temporary halt of the cranial rhythmic impulse (stillpoint) was decreased when compared to prestillpoint MSNA (p < 0.01). During the CV4 sham and control trials MSNA was not different between CV4 time-matched measurements (p > 0.05). Moreover, the change in MSNA prestillpoint to stillpoint during the CV4 trial was different compared to the CV4 sham and control trials (p < 0.05). However, this change in MSNA was similar between the CV4 sham and control trials (p > 0.80). CONCLUSIONS: The current study is the first to demonstrate that cranial manipulation, specifically the CV4 technique, can alter sleep latency and directly measured MSNA in healthy humans. These findings provide important insight into the possible physiologic effects of cranial manipulation. However, the mechanisms behind these changes remain unclear. PMID- 15750370 TI - Perception of bodily sensation as a predictor of treatment response to acupuncture for postoperative nausea and vomiting prophylaxis. AB - OBJECTIVES: Conditions associated with the response to acupuncture treatment remain widely elusive. This study aimed to identify predictors of treatment response in patients undergoing acupuncture treatment for prevention of postoperative nausea and vomiting (PONV). An intervention-specific questionnaire was developed to assess perception of bodily sensations (PBS) in patients undergoing acupuncture treatment. DESIGN: Exploratory study in addition to an observer-blinded randomized controlled trial. SETTINGS/LOCATION: The departments of anesthesiology and of gynecology, University of Heidelberg, Germany. SUBJECTS: Two hundred and twenty (220) female patients scheduled for breast or gynecologic surgery. INTERVENTIONS: True acupuncture on point P6 (Neiguan) or placebo/sham acupuncture on a virtual point 1 cm proximal and lateral to P6. OUTCOME MEASURES: Of 220 female patients scheduled for breast or gynecologic surgery who were included in the trial, 202 completed the newly developed PBS questionnaire before the intervention. The main outcome was incidence of PONV in relationship with PBS and the kind of acupuncture applied. Logistic regression analyses were performed to identify predictors of treatment response for those receiving surgery (n = 195). RESULTS: High PBS predicted low treatment response in patients receiving breast surgery (odds ratio [OR] = 1.6; p = 0.039 for each 10-point increase). The PBS questionnaire showed good item difficulty, internal consistency, and divergent validity. Treatment response in patients receiving gynecologic surgery was predicted by the kind of acupuncture applied (OR = 0.4; p = 0.027), whereas PBS played no role in this group. CONCLUSIONS: The incidence of PONV after breast surgery seems to be influenced by some psychological mechanisms. In this group of patients, acupuncture might be more effective in patients who have low PBS. PMID- 15750369 TI - A randomized trial investigating a chiropractic manual placebo: a novel design using standardized forces in the delivery of active and control treatments. AB - OBJECTIVES: To evaluate the proposed manual placebo in terms of success in blinding patients to treatment group assignment and outcomes between the treatment groups. DESIGN: Randomized controlled trial. SETTING: A chiropractic college research clinic in the midwestern United States. SUBJECTS: One hundred and eleven (111) individuals aged 18 years and over with subacute or chronic lowback pain. INTERVENTIONS: The active treatment consisted of flexion distraction chiropractic manipulation and trigger point therapy and the control treatment of sham manipulation and effleurage; both groups received eight treatments over a 3-week period. The application of prescribed ranges of biomechanical forces for each treatment was standardized using specialized computerized equipment. "Nontreatment" aspects of the clinical encounter were to be standardized across groups. A primary clinician blinded to treatment assignment provided interpersonal interactions and treating clinicians delivered treatments with a minimum of interaction. OUTCOME MEASURES: The accuracy of the patient's perception of group assignment at visit 4 and the mean change in the Pain Disability Index (PDI) over the treatment period were the primary outcome variables. RESULTS: Patients in the control group were more likely to perceive their treatment assignment accurately than those in the active group (78% versus 54%, respectively). Patients in both treatment groups improved on the PDI and the Roland-Morris Questionnaire; there were no significant differences in improvement between the groups. Age, gender, prior chiropractic experience and expectation of treatment at baseline had no effect on outcomes. CONCLUSIONS: Patients in the control group were not successfully blinded; however, patients' perceptions of treatment group assignment did not significantly affect outcomes. The clinically significant improvement in both groups, independent of patient or clinician expectations, suggests the presence of therapeutic factors common to both groups, other than biomechanical force. Further studies examining other aspects of the clinical encounter, considered separately from biomechanical force, are warranted before arbitrarily designating any intervention as a "placebo." PMID- 15750371 TI - Effect of electroacupuncture on bronchial asthma induced by ovalbumin in rats. AB - INTRODUCTION: Asthma is a worldwide disabling chronic inflammatory airway disease characterized by an intense eosinophilic inflammatory infiltrate on bronchial mucous membranes. Among the complementary therapeutic approaches to asthma, acupuncture has been widely used. OBJECTIVE: Here we used a rat pulmonary hypersensitivity experimental model that mimics human asthma in order to address whether electroacupuncture (EA) treatment could reduce the inflammatory process. MATERIALS AND METHODS: Experimental animals were divided in four groups: control (C), immobilized (I), sham-acupuncture (SA), and acupuncture (A). All rats were sensitized with heat-solidified hen egg white implant. Using clinical acupuncture points, EA treatment began 2 days after antigen priming and was repeated on alternate days for 2 weeks. Subsequently, animals were challenged by inhalation with aggregated ovalbumin and sacrificed 24 hours later when blood samples, bronchoalveolar lavage (BAL), and lungs were collected. RESULTS: Histopathologic analyses showed that peribronchial and perivascular inflammatory cell infiltrates were significantly lower in group A compared to groups SA and I (shown to be similar to group C). Furthermore, BAL total cell count and percentage of polymorphonuclears (as well as the differential counts of neutrophils and eosinophils) were significantly reduced in group A compared to group I. Corsticosterone plasma levels were similar in all groups. CONCLUSIONS: Taken together these results show that EA efficiently diminishes the bronchial immune mediated inflammation induced in rats and that this effect is dependent on the choice of specific acupoints. PMID- 15750372 TI - Electroacupuncture attenuates inflammation in a rat model. AB - BACKGROUND: Acupuncture has traditionally been used in China and is being increasingly applied in Western countries to treat a variety of conditions, including inflammatory disease. However, clinical trials investigating the effectiveness of the anti-inflammatory effects of acupuncture have yielded inconsistent results, and the underlying mechanisms of acupuncture-produced anti inflammation are unclear. OBJECTIVE: To evaluate the effectiveness of electroacupuncture (EA) on inflammation in a rat model. MATERIALS AND METHODS: Four experiments were conducted on male Sprague-Dawley rats (n = 8-9 per group). Inflammation was induced by injecting complete Freund's adjuvant (CFA) subcutaneously into the plantar surface of one hind paw of the rat. Experiment 1: To determine the effect of EA (10 and 100 Hz) versus sham treatment on inflammation. Experiment 2: To investigate the involvement of the adrenal glands on the effect of EA treatment using adrenalectomized (ADX) rats. Experiment 3: To determine the effects of EA on plasma levels of corticosterone. Experiment 4: To determine the effects of EA treatment versus immobilization on such stress indicators as heart rate and blood pressure. RESULTS: At 10 Hz EA significantly reduced CFA-induced hind paw edema. The effect was partially blocked in the ADX rats. EA significantly increased plasma levels of corticosterone but produced no noticeable signs of stress. CONCLUSION: At 10 Hz but not 100 Hz, EA suppresses inflammation by activating the hypothalamus-pituitary-adrenal axis (HPA) and the nervous system. PMID- 15750373 TI - Antioxidant and protective effects of Amrit Nectar tablets on adriamycin- and cisplatin-induced toxicities. AB - OBJECTIVES: Maharishi herbal food supplements have been shown to inhibit the growth of mammary tumors and reduce free radical-mediated injuries. The purpose of this investigation is to evaluate the effects of aqueous and alcoholic extracts of Amrit Nectar tablets on rat liver microsomal lipid peroxidation and compare to other antioxidants. The protective effects of dietary Amrit Nectar tablets (MA-7; containing 38 herbs) on cisplatin-induced changes in glutathione (GSH) and glutathione-S-transferase (GST) activity in rat liver and kidney, and Adriamycin (Pharmacia S.p.A, Milan, Italy)-induced mortalities in mice were also investigated. RESULTS: Both aqueous and alcoholic extracts of MA-7 were more potent than other antioxidants tested under our experimental conditions. Adriamycin (15 mg/kg, intraperitoneally) caused 60% mortality during the period of 4 weeks in CDF1 mice. Dietary MA-7 (0.7%) treatment decreased the mortality to 20%. Dietary MA-7 (0.7%) supplementation with cisplatin treatment reversed the effects of cisplatin on liver and kidney GSH and GST activity. CONCLUSIONS: These results indicate that MA-7 is a powerful antioxidant. MA-7 supplementation with Adriamycin and cisplatin treatment may protect against their toxicities. PMID- 15750374 TI - Ginger extract components suppress induction of chemokine expression in human synoviocytes. AB - INTRODUCTION: Ginger has a long history of medicinal use, particularly as an anti inflammatory agent for a wide variety of diseases such as arthritis. Suppression of inflammation in arthritis is attributed to suppression of proinflammatory cytokines and chemokines produced by synoviocytes, chondrocytes, and leukocytes. OBJECTIVE: This study aimed to elucidate the effect of a combination ginger extract and its individual components on chemokine expression in human synoviocytes. METHODS: Human synoviocytes were incubated with 100 microg/mL combination ginger extract (GE) of Alpinia galanga (AG) and Zingiber officinale (ZO); AG extract alone; ZO extract alone; or control media, for 1 hour at 37 degrees C, 5% CO2. Cells were next activated with 1 ng/mL of tumor necrosis factor alpha (TNF-alpha) for 1 hour to determine macrophage chemotactic factor (MCP-1) and interferon-gamma activated protein (IP-10) mRNA levels using reverse transcriptase polymerase chain reaction (RT-PCR). Secreted MCP-1 and IP-10 were quantified by enzyme-linked immunosorbent assay (ELISA) following a 24 hour incubation period. RESULTS: The GE combination was consistently more effective in decreasing chemokine mRNA and chemokine secreted protein levels than its individual components ZO or AG. In comparison, ZO was more effective than AG in suppressing chemokine expression. CONCLUSION: The present study demonstrates that GE inhibits chemokine expression, and that the combination of ZO and AG components acts synergistically. This ginger formulation may be useful for suppressing inflammation due to arthritis. PMID- 15750375 TI - The homeopathic preparation Vertigoheel versus Ginkgo biloba in the treatment of vertigo in an elderly population: a double-blinded, randomized, controlled clinical trial. AB - OBJECTIVE: Alternative medical practices are common in the treatment of vertigo. This study compared the effects of Ginkgo biloba treatment with the homeopathic remedy Vertigoheel (Biologische Heilmittel Heel GmbH, Baden-Baden, Germany). DESIGN: Randomized, double-blinded, parallel group study. SUBJECTS: One hundred and seventy (170) patients, ages 60-80 years, with atherosclerosis-related vertigo. INTERVENTIONS: Patients were randomly allocated to receive treatment with either Vertigoheel (n = 87) or G. biloba (n = 83). OUTCOME MEASURES: The results were analyzed for the non-inferiority of Vertigoheel to G. biloba on the combined endpoint of changes from baseline to week 6 in dizziness score (assessed by questionnaire), frequency, duration, and intensity of vertigo episodes (recorded in patient diaries). RESULTS: Both treatments improved vertigo status. From a baseline mean value of 26.1 +/- 5.2 (on a 50-point scale) in the Vertigoheel group, the dizziness questionnaire score improved by -10.6 +/- 10.0, and by -10.7 +/- 9.0 from 25.8 - 4.7 in the G. biloba group. Statistical analysis of this endpoint showed that Vertigoheel was not inferior to G. biloba. The 95% confidence interval for the difference between treatment did not reach the inferiority threshold of 0.36 at any of the time points tested. The results were supported by the results of a line walking test, Unterberger's stepping test, and patient and physician global assessments of therapeutic effect. Both treatments were well tolerated. CONCLUSIONS: Vertigoheel is an appealing alternative to established G. biloba therapy for atherosclerosis-related vertigo. PMID- 15750376 TI - Bovine cartilage, coenzyme Q10, and wheat grass therapy for primary peritoneal cancer. AB - The accepted postsurgical first-line therapy for primary peritoneal cancer has been a regime of chemotherapy. This paper reports the case of an 89-year-old female who refused chemotherapy but accepted a nutritional alternative. Results after more than 4 years of the nutritional regime have been encouraging with regards to objective and subjective measures. PMID- 15750377 TI - Acupuncture treatment of compression neuropathy of the radial nerve: a single case report of "Saturday Night Palsy". AB - PURPOSE: Evidence that acupuncture is effective for any type of motor nerve injury is limited to case reports and case series but these findings indicate benefit. Observation that the radial nerve has the most rapid recovery of all peripheral nerves suggests that acupuncture might benefit treatment of "Saturday Night Palsy," a syndrome of radial-nerve compression. TREATMENT: A 41-year-old female with a 1-week history of inability to write or extend the right wrist received 1 acupuncture treatment utilizing the Lung and Large Intestine meridians in the forearm, with the 2 meridians interconnected using the Luo and Yuan points. A cockup wrist splint was then applied. CONCLUSIONS: Wrist motion with gravity neutralized returned immediately after treatment. As the day progressed, the patient reported increasing strength in wrist and finger extension. The next day, the patient cancelled the second acupuncture treatment, as her hand had recovered. Examination 4 months later revealed normal wrist and finger extension, sensation, and return of the brachioradialis reflex. The patient was symptom-free 1-year postinjury. Acupuncture potentially facilitates recovery and may enhance treatment of peripheral motor nerve injury. PMID- 15750378 TI - Biophotonics in the infrared spectral range reveal acupuncture meridian structure of the body. AB - This study shows, for the first time, evidence of the existence of the acupuncture meridian structure in the human body. After moxibustion (or similar light stimulation) of the body in the 3-5 microm range, "light channels" appear on the body, which appear to be identical to what are known as meridians in all textbooks of Traditional Chinese Medicine. These findings appear not only to confirm the existence of acupuncture meridians, but they also open a new window on understanding the energy transfer dynamics of the human body. Furthermore, it is likely that living matter is not in the ground state, but permanently electronically excited. PMID- 15750379 TI - Does acupuncture help stroke patients become more independent? AB - This short speculative report describes the outcome of three studies looking at the effect of acupuncture on stroke recovery and the subsequent place of residence of the subjects entered. It is not a systematic review and does not endeavor to provide comprehensive data on the effect of acupuncture on post stroke recovery. Our observations demonstrate that patients may be more likely to remain independent and in their own homes one year post stroke if they receive acupuncture. This conclusion is supported by our study and two previous trials. It may be that acupuncture improves post-stroke perception, thereby enhancing independence. PMID- 15750380 TI - Square pegs and round holes? A review of economic evaluation in complementary and alternative medicine. AB - INTRODUCTION: Economic evaluation, linking the costs and consequences of an intervention to indicate the potential benefits of alternative interventions, is becoming established as one of the core tools for decision making in health care. As knowledge of the safety and effectiveness of complementary and alternative medicine (CAM) interventions increases, economic evaluation within CAM has a heightened significance. OBJECTIVE: To explore whether the present framework for economic evaluation fits CAM and what modifications if any are needed for its application. DESIGN: Systematic review. METHODS: A comprehensive search of four databases was undertaken (NHS Economic Evaluation Database, AMED, MEDLINE, CINAHL). Studies were included if they took the form of a comparative analysis of costs and consequences of a CAM treatment and were written in English. Each study was reviewed using a set of methodological questions to judge their quality as economic evaluations. RESULTS: A total of 19 studies were identified, of which 9 were cost-effectiveness studies, 7 cost-consequence studies, 2 cost-minimization studies, and 1 cost-benefit analysis. Seventeen (17) of the studies involved CAM treatments being used alongside mainstream or conventional treatments. The majority of the treatments aimed to alleviate pain, including chronic pain, back pain, neck pain, and migraine. Only a small minority of studies addressed wider outcomes of particular relevance to CAM disciplines. Nine (9) adopted a service provider perspective only, 7 included wider sickness absence costs and 3 patient costs. Only 1 study included costs to relatives. The quality of the cost and benefit dimensions of the studies was mixed. CONCLUSIONS: A CAM sensitive approach to economic evaluation is required. This needs to include a focus on outcomes that explore the range of effects of CAM treatment, an exploration of the client's perspective and not just that of the service provider and study designs that facilitate the individualized practitioner approach so central to CAM treatment. PMID- 15750381 TI - Sudarshan Kriya yogic breathing in the treatment of stress, anxiety, and depression: part I-neurophysiologic model. AB - Mind-body interventions are beneficial in stress-related mental and physical disorders. Current research is finding associations between emotional disorders and vagal tone as indicated by heart rate variability. A neurophysiologic model of yogic breathing proposes to integrate research on yoga with polyvagal theory, vagal stimulation, hyperventilation, and clinical observations. Yogic breathing is a unique method for balancing the autonomic nervous system and influencing psychologic and stress-related disorders. Many studies demonstrate effects of yogic breathing on brain function and physiologic parameters, but the mechanisms have not been clarified. Sudarshan Kriya yoga (SKY), a sequence of specific breathing techniques (ujjayi, bhastrika, and Sudarshan Kriya) can alleviate anxiety, depression, everyday stress, post-traumatic stress, and stress-related medical illnesses. Mechanisms contributing to a state of calm alertness include increased parasympathetic drive, calming of stress response systems, neuroendocrine release of hormones, and thalamic generators. This model has heuristic value, research implications, and clinical applications. PMID- 15750382 TI - New initiative--Interuniversity European Union project: distance learning program in integrated health sciences. PMID- 15750383 TI - "No language restrictions" in database searches: what does this really mean? AB - The aim of this study was to investigate the coverage of non-English journals by MEDLINE((R)) and EMBASE, the two major biomedical databases used for identifying studies for possible inclusion in systematic reviews and meta-analyses. A series of searches were conducted to compare the coverage of journals in languages other than English. The results were compared against listings in Ulrich's Periodicals Directory, an authoritative source of information on periodicals published in more than 200 countries. This study has highlighted the existence of a database coverage bias, in terms of the systematic exclusion of journals from certain countries and/or in certain languages. Searching that relies only on English language databases may result in failure to find many relevant studies published in languages other than English, irrespective of the research question and the avoidance of any language restrictions. PMID- 15750384 TI - Complementary and alternative medicine in the scientific literature. AB - OBJECTIVE: To analyze the frequency and typology of scientific papers on complementary and alternative medicine (CAM) published in the period 1997-2002. METHODS: We performed a systematic literature search of MEDLINE in order to identify all articles dealing with CAM, in the human setting, published from January 1, 1997 to December 31, 2002. Our attention was focused on randomized controlled trials conducted on CAM and on the impact factor (IF) of the journals in which CAM articles were published. RESULTS: During the period 1997-2002, a total of 20,209 articles about CAM were published, representing the 0.7% of the total number of MEDLINE-listed articles. Approximately 50% of CAM articles published in 1996-2002 appeared on journals with no IF. The proportion of randomized clinical trials was 7.6% of total CAM articles. CONCLUSIONS: There is an upward trend in the number of CAM articles but the number of experimental studies is low and over half of articles are published in journals without an IF. We believe there is an urgent need to conduct rigorous research in the field of CAM in order to support, for the need of the public, an evidence-based approach to these therapies. PMID- 15750385 TI - From asthma to Alzheimer's: Cochrane vitamin reviews cover an array of topics. AB - In the current issue of the Cochrane Library (Issue 3, 2004), there are more than 25 systematic reviews addressing the use of vitamins in the prevention or treatment of disease. Vitamins have been a mainstay of health since their discovery in the early 1900s, so a question arises as to when using vitamins is considered complementary as opposed to conventional medical practice. Complementary or alternative vitamin use encompasses those uses that are outside of accepted medical practice. The use of vitamin C to prevent scurvy is accepted medical practice. The use of vitamin C as a therapeutic adjunct to asthma is not. This paper summarizes 15 Cochrane systematic reviews on the complementary use of vitamins for an array of conditions including the common cold, Alzheimer's disease, asthma, chemotherapy-induced mucositis, and depression. PMID- 15750387 TI - Altered bone mass in children at diagnosis of Crohn disease: a pilot study. AB - OBJECTIVES: Recent studies have indicated that bone mineral density is reduced in children with inflammatory bowel disease. The exact cause of this reduction is unclear, but it is often attributed to corticosteroid use. This study examined the prevalence of reduced bone mass in otherwise healthy children newly diagnosed with Crohn disease without previous corticosteroid exposure. METHODS: Eighteen steroid-naive children newly diagnosed with Crohn disease underwent dual energy x ray absorptiometry. Disease activity, growth and pubertal development, nutritional assessment and bone mass measurements were recorded. z scores were adjusted for bone age. RESULTS: Five of the 18 patients (28%) had a total bone mineral density z score less than -1 (one had a z score less than -2). Ten (56%) subjects had lumbar spine bone mineral density z scores less than -1 (two had z score less than -2). The subjects had significantly reduced mean lumbar spine bone mineral density z scores (P = 0.002). Delayed pubertal development correlated with whole body bone mineral density z scores (r = 0.64; P = 0.004). Most subjects were not meeting United States recommended dietary allowances for daily intake of calcium, vitamin D and total calories. The majority of subjects were not participating in weight-bearing physical activity. CONCLUSION: Decreased bone mass is common in steroid naive children newly diagnosed with Crohn disease. Crohn disease appears to contribute to impaired bone mass independent of corticosteroid therapy. PMID- 15750388 TI - Diagnosis: Condyloma acuminatum. PMID- 15750389 TI - Sex and the course of HIV infection in the pre- and highly active antiretroviral therapy eras. AB - We reviewed the available literature on the potential effects of sex on the course of HIV infection and found that there is little evidence for sex differences in the rate of disease progression in the pre-highly active antiretroviral therapy (HAART) and HAART era. Compared to men, women appeared to have lower HIV RNA levels and higher CD4 cell counts shortly after infection with HIV, but studies were inconclusive regarding whether these differences diminish over time. Differences in viral load or CD4+ cell count might cause women to delay initiation of HAART. Nonetheless, we found no substantial sex difference in the benefit of antiretroviral therapy. The studies we reviewed failed to find any harmful effect of pregnancy on HIV disease progression. With the availability of effective antiretroviral agents, HIV-infected women have increasingly decided to have children. Conflicting results exist on the effect of HAART on regression of cervical intra-epithelial neoplasia (CIN). Unlike CIN, invasive cervical cancer has not been found to be much higher in HIV-infected women than in HIV-uninfected women. Although publication bias cannot be ruled out, published studies suggest higher rates of adverse events among HIV-infected women on therapy as compared to men. As more pharmacological agents are developed, it is especially important that potential sex differences in pharmacodynamics are assessed. The relationship between metabolic abnormalities, changes in body habitus, and endocrine perturbations has not been extensively studied. Whether sex differences are due to unalterable genetic factors or social and environmental conditions, it is imperative that all HIV-infected individuals have equal access to interventions that can slow disease progression. PMID- 15750390 TI - An MDR1-3435 variant is associated with higher plasma nelfinavir levels and more rapid virologic response in HIV-1 infected children. AB - OBJECTIVE: The multidrug-resistance transporter gene (MDR1) encoding for P glycoprotein (P-gp) and genes encoding for isoenzymes of cytochrome P450 (CYP) have an important role in transport and metabolism of antiretroviral agents. This research examined the impact of single nucleotide polymorphisms (SNP) of MDR1 and CYP genes on nelfinavir and efavirenz pharmacokinetics and the response to highly active antiretroviral therapy (HAART) in HIV-1 infected children. METHODS: Seventy-one HIV-1-infected children from PACTG 382 receiving nelfinavir, efavirenz and one or two nucleoside reverse transcriptase inhibitors had genomic DNA from PBMC evaluated for MDR1 and CYP SNP by real-time PCR. Plasma drug concentrations, CD4 lymphocyte counts and HIV-1 RNA were measured during HAART. RESULTS: The frequencies of C/C, C/T and T/T genotypes in the MDR1-3435-C-->T polymorphisms were 44% (n = 31), 46% (n = 33) and 10% (n = 7), respectively. Ninety-one percent of children with the C/T genotype reached plasma HIV-1 RNA < 400 copies/ml by week 8 compared to 59% of children with the C/C genotype (P = 0.01). Children with the C/T genotypes had higher 8 h postdose concentration (P = 0.02) and lower clearance rate (P = 0.04) for nelfinavir compared to those with the C/C genotype. The seven children with the T/T genotype had nelfinavir pharmacokinetics and virologic response similar to those with the C/C genotype. No compensatory polymorphisms were observed between MDR1 and CYP genotypes. CONCLUSIONS: HIV-1 infected children with the MDR1-3435-C/T genotype had more rapid virologic responses to HAART at week 8 with higher plasma nelfinavir concentrations compared to those with the C/C genotype. These findings suggest that P-gp may play an important role in the pharmacokinetics and virologic response to HAART containing nelfinavir. PMID- 15750391 TI - Increased levels of galactose-deficient IgG in sera of HIV-1-infected individuals. AB - BACKGROUND: The IgG from sera of patients with chronic inflammatory diseases of autoimmune character or some chronic microbial infections is frequently deficient in galactose on N-linked glycans. However, this phenomenon has not been investigated at length in human viral infections. OBJECTIVES: To evaluate the glycosylation of serum IgG in HIV-1-positive patients. METHODS: Psathyrella velutina lectin was used in enzyme-linked immunosorbent and Western blot assays to determine glycosylation. In addition, gas-liquid chromatography and mass spectrometry were utilized to confirm the galactose deficiency observed in the lectin-binding assays. RESULTS: HIV-1-infected individuals had significantly higher levels of galactose-deficient IgG than healthy controls. In fact, the galactose deficiency of the N-linked glycans observed in other diseases was even more profound in HIV-1 infection. This deficiency was primarily restricted to IgG when total serum glycoproteins were evaluated and IgG1 was the subclass most affected in all patients. Also, a significant increase in lectin binding was observed on IgG2 and IgG4 from HIV-1-positive females compared with HIV-1 negative females. CONCLUSIONS: Identification of deficient galactosylation of serum IgG from HIV-1-infected patients extended the spectrum of diseases in which this phenomenon has been observed. In addition, the results suggest yet another aspect of immune dysfunction as a result of HIV-1 infection. PMID- 15750392 TI - HIV-1 resistance genotyping on dried serum spots. AB - OBJECTIVE: To assess the feasibility of HIV-1 group M resistance genotyping on dried serum spots, by testing samples from previously untreated patients, patients on treatment, and patients having stopped treatment, representing a wide genetic diversity panel. METHODS: Serum samples from 62 HIV-1-infected Caucasian and African patients, with viral load values from 715 copies/ml to more than 750,000 copies/ml, were deposited on filter paper. After elution and RNA extraction, nested RT-PCR was used to amplify the protease and RT regions of the pol gene. Resistance sequencing was performed on all the protease and RT amplicons. The sequences obtained for resistance genotyping were used for subtyping by phylogenetic analysis. RESULTS: Amplification was successful in the protease region in 53/62 cases (85.5%) and in the RT region in 51/62 cases (82.3%). All samples with viral loads of at least 5 Log (17 of 62) were successfully amplified in both the RT and protease regions. Of the 29 samples with viral loads between 4 Log and 5 Log, 28 (97%) were amplified in the RT region and 25 (86%) in the protease region. The detected mutations were in keeping with the treatment status. Marked natural polymorphism was observed in the protease region, but no major consequences were deduced in terms of resistance. The results showed a broad diversity of the panel, including subtype B (n = 36) and non B or recombinant forms (n = 20). CONCLUSION: Our results show the feasibility of this dried serum spot method for monitoring resistance to antiretroviral drugs and the molecular epidemiology of HIV diversity. The simplicity of sample preparation, storage and transport potentially makes this an importance tool for individual and epidemiological monitoring throughout the world. PMID- 15750393 TI - Incidence and risk factors for immune reconstitution inflammatory syndrome during highly active antiretroviral therapy. AB - BACKGROUND: There is little systematic information regarding the immune reconstitution inflammatory syndrome (IRIS). OBJECTIVE: To determine the incidence, risk factors, and long-term outcome of IRIS in HIV-infected patients receiving highly active antiretroviral therapy (HAART) who were coinfected with one of three common opportunistic pathogens. DESIGN: A retrospective cohort identified through a city-wide prospective surveillance program. METHODS: A retrospective chart review was performed for 180 HIV-infected patients who received HAART and were coinfected with Mycobacterium tuberculosis, Mycobacterium avium complex, or Cryptococcus neoformans between 1997 and 2000. Medical records were reviewed for baseline demographics, receipt and type of HAART, response to antiretroviral therapy, development of IRIS, and long-term outcome. RESULTS: In this cohort, 31.7% of patients who received HAART developed IRIS. Patients with IRIS were more likely to have initiated HAART nearer to the time of diagnosis of their opportunistic infection (P < 0.001), to have been antiretroviral naive at time of diagnosis of their opportunistic infection (P < 0.001), and to have a more rapid initial fall in HIV-1 RNA level in response to HAART (P < 0.001). CONCLUSIONS: IRIS is common among HIV-infected persons coinfected with M. tuberculosis, M. avium complex, or C. neoformans. Antiretroviral drug-naive patients who start HAART in close proximity to the diagnosis of an opportunistic infection and have a rapid decline in HIV-1 RNA level should be monitored for development of this disorder. PMID- 15750394 TI - Brain deposition of beta-amyloid is a common pathologic feature in HIV positive patients. AB - BACKGROUND: We planned to analyze the prevalence and distribution of beta-amyloid deposition in the HIV+ brain in the HAART era. Our working hypothesis is that long term survival, aging and the secondary effects of HAART may contribute to increased beta-amyloid accumulation in this patient population. METHODS: Paraffin embedded archival brain autopsy tissues were assessed by immunocytochemistry for beta-amyloid. Detailed in-vivo neuro-behavioral assessments and ApoE genotyping were available for a subset of the studied population. RESULTS: Immunoreactivity with the antibodies 4G8 and 6E10 was found predominantly in neuronal soma and dystrophic axonal processes. Extracellular, often perivascular plaques were also identified in many cases. CONCLUSIONS: We propose that prolonged HAART and aging may contribute to an overall increase in amyloid deposition, potentially mediated by inhibition of insulin degradation enzyme (IDE) or disruption of the axonal transport of the amyloid precursor protein. PMID- 15750395 TI - Evaluating the potential impact of vaginal microbicides to reduce the risk of acquiring HIV in female sex workers. AB - OBJECTIVES: The following questions were addressed: would the introduction of vaginal microbicides substantially reduce the risk of female sex workers (FSWs) acquiring HIV? Which factor would it be most important to maximize, microbicide efficacy or microbicide use? What level of microbicide efficacy and use would be necessary to counterbalance a possible reduction in condom use? DESIGN: Mathematical modeling, with parameter estimations from available literature. METHODS: Risk equations were developed and Monte Carlo simulations were performed to model a FSW's daily risk of HIV acquisition currently, and after, microbicide introduction. Uncertainty and sensitivity analyses were used as well as tornado plots for two ranges of microbicide efficacy (30-50%) and (50-80%). Risk was estimated for FSWs whose clients sometimes (10-50%) use condoms, and those whose clients never use condoms. An analytical threshold for which reducing condom use increases risk was estimated. RESULTS: For both groups of FSWs, daily risk would decrease by approximately 17% or approximately 28% using 30-50% or 50-80% effective microbicides, respectively. Increasing microbicide use would have greater impact on reducing risk than increasing microbicide efficacy. The microbicide efficacy and usage required to ensure that 'condom replacement' does not increase a FSW's risk of acquiring HIV was calculated. CONCLUSIONS: Microbicides could substantially reduce FSWs' risk of acquiring HIV; absolute decrease in risk would be greatest in high-prevalence regions. The public health impact of microbicides will depend upon usage and efficacy. Even if the microbicides that become available are only low-to-moderately effective, the probability that risk in FSWs will increase (due to replacing condoms with microbicides) is low. PMID- 15750397 TI - Mode of infant feeding and HIV infection in children in a program for prevention of mother-to-child transmission in Uganda. AB - OBJECTIVE: To evaluate the impact of different modalities of infant feeding on HIV transmission in children in a prevention of mother-to-child transmission (PMTCT) program in an urban hospital in Uganda. METHODS: HIV-infected pregnant women in the PMTCT program at St Francis Hospital Nsambya, Kampala were offered the chance to participate in the study. Short-course antiretroviral regimens were provided and formula feeding offered free of charge for women choosing not to breastfeed. Mother-infant pairs were followed until 6 months postpartum. HIV status in children was assessed at week 6 and month 6. For the analyses, mother infant pairs were classified into three groups according to the mode of infant feeding: exclusive formula feeding (EFF), exclusive breastfeeding (EBF) and mixed feeding (MF). RESULTS: A total of 306 children were enrolled. Transmission rates were 8.9% at week 6 (3.4% in the EFF group, 11.2% in the EBF group, 17.1% in the MF group) and 12.0% at month 6 (3.7% in the EFF group, 16.0% in the EBF group, and 20.4% in the MF group). The EBF and MF groups were associated with a significantly higher risk of HIV transmission than the EFF group. No significant risk difference was observed between the EBF and the MF groups. CONCLUSIONS: HIV transmission rates were significantly lower in formula-fed infants in comparison with both exclusively breastfed and mixed-fed infants. Transmission through breastfeeding seems to occur mainly in the first weeks after delivery. PMID- 15750396 TI - Efficacy of a brief case management intervention to link recently diagnosed HIV infected persons to care. AB - OBJECTIVE: The Antiretroviral Treatment Access Study (ARTAS) assessed a case management intervention to improve linkage to care for persons recently receiving an HIV diagnosis. METHODS: Participants were recently diagnosed HIV-infected persons in Atlanta, Baltimore, Los Angeles and Miami. They were randomized to either standard of care (SOC) passive referral or case management (CM) for linkage to nearby HIV clinics. The SOC arm received information about HIV and local care resources; the CM intervention arm included up to five contacts with a case manager over a 90-day period. The outcome measure was self-reported attendance at an HIV care clinic at least twice over a 12-month period. RESULTS: A higher proportion of the 136 case-managed participants than the 137 SOC participants visited an HIV clinician at least once within 6 months [78 versus 60%; adjusted relative risk (RR(adj)), 1.36; P = 0.0005) and at least twice within 12 months (64 versus 49%; RR(adj), 1.41; P = 0.006). Individuals older than 40 years, Hispanic participants, individuals enrolled within 6 months of an HIV-seropositive test result and participants without recent crack cocaine use were all significantly more likely to have made two visits to an HIV care provider. We estimate the cost of such case management to be 600-1200 US dollars per client. CONCLUSION: A brief intervention by a case manager was associated with a significantly higher rate of successful linkage to HIV care. Brief case management is an affordable and effective resource that can be offered to HIV infected clients soon after their HIV diagnosis. PMID- 15750398 TI - Biphasic decline of CD4 cell count during scheduled treatment interruptions. AB - In 115 patients whose scheduled treatment interruptions (STI) lasted 24 weeks, the CD4 cell count declined by a median of 30 cells/ml/week during the first 4 weeks, compared with 3 cells/ml/week during the next 20 weeks. In multivariate regression, a pronounced early fall in CD4 cells correlated with a higher CD4 cell count at the start of STI, with more gain in CD4 cells during antiretroviral treatment preceding STI, and with a higher viral load at week 4. PMID- 15750399 TI - Generic antiretroviral efficacy in AIDS-associated Kaposi's sarcoma in sub Saharan Africa. AB - Generic antiretroviral drugs are pivotal in the implementation of WHO's '3 by 5' programme. However, clinical experience with generics in sub-Saharan Africa is insufficiently documented. We report on 50 patients with HIV-associated Kaposi's sarcoma treated with generic fixed-dose highly active antiretroviral therapy. At 52 weeks, 74% achieved an undetectable viral load of < 50 copies/ml, 86% achieved < 400 copies/ml, and a 3.1 log10 decline from baseline. Side-effects were minimal. The outcomes support the use of generic antiretroviral therapy. PMID- 15750400 TI - Decreasing incidence of pregnancy by decreasing CD4 cell count in HIV-infected women in Cote d'Ivoire: a 7-year cohort study. AB - In a cohort study of women of childbearing age in Abidjan, Cote d'Ivoire, we followed 473 HIV-infected women for 1551 person-years, and found that the incidence of pregnancy and livebirth decreased with decreasing CD4 cell counts. This has consequences in terms of scaling-up strategies for highly active antiretroviral therapy (HAART). Women who need HAART will be less likely than those who do not to be recruited into prenatal care facilities. PMID- 15750401 TI - Lopinavir/ritonavir maintenance monotherapy after successful viral suppression with standard highly active antiretroviral therapy in HIV-1-infected patients. PMID- 15750402 TI - Investigation of HIV-1 transmission events by phylogenetic methods: requirement for scientific rigour. PMID- 15750403 TI - Successful desensitization of enfuvirtide-induced skin hypersensitivity reaction. PMID- 15750404 TI - Strategies for Leadership: Nursing Leadership for Patient Safety. PMID- 15750406 TI - Moral distress at 3 am. PMID- 15750408 TI - Nurse executive's legal primer. PMID- 15750411 TI - Focus group on licensed practical nurse scope of practice at National Council of State Boards of Nursing. PMID- 15750409 TI - What's a nurse to do?: How the Health Insurance Portability and Accountability Act of 1996 impacts a nurse's (or any other healthcare provider's) ex parte discussion of protected health information in medical malpractice cases. PMID- 15750412 TI - The business of medicine, how real it is. PMID- 15750413 TI - Reactions to biomaterials: the good, the bad, and ideas for developing new therapeutic approaches. PMID- 15750414 TI - Course of the masseteric nerve in masseter muscle. AB - The masseter muscle is often exploited by plastic and reconstructive surgeons in transposition operations to correct facial palsy. In benign masseteric hypertrophy, a neurectomy induces atrophy of the muscle and therefore a slender face. A clear understanding of the course of the masseteric nerve is essential in maneuvering the masseter muscle. Detailed dissection was performed on 48 head halves of Korean cadavers, and the inside of each masseter muscle was explored under the microscope with x10 magnification. The masseteric nerve ran anteriorly and inferiorly between the deep and the middle layers. The nerve was observed at 33+/-5.6 mm from the inferior border of the muscle on the anterior third vertical line of the masseter muscle and at 47+/-5.5 mm in the posterior third. The closest distance between the main nerve and the gonion was 32+/-4.1 mm. Approximately two thirds (69.6%) of the masseteric nerve intersected the artery. The safe limit is within 47 mm from the inferior border of the muscle on the posterior third vertical line and within radial 32 mm from the gonion. PMID- 15750415 TI - How cranial could the sternocleidomastoid muscle be split? AB - The sternocleidomastoid (SCM) muscle, its anatomy, and its possible uses in head and neck reconstruction have been discussed, especially during the last 2 decades. The viability of the muscle and the skin over it depends on the vascular anatomy of the SCM and the anatomical heads-anterior or sternal and posterior or clavicular-that could enable splitting of the SCM muscle or myocutaneous flap. The authors have investigated the nourishment pattern of the SCM muscle to find out whether there was an independent blood supply to the each head of the muscle anterior or sternal and posterior or clavicular-and whether the anatomical discrimination of the heads was congruent with the vascular pattern of the heads. The distinct vascular pattern of the two heads was determined. The occipital artery supplied all the muscle, and the first minor pedicle of the muscle originating from the superior thyroid artery supplies the clavicular head. Surgically, the heads could be split cranially until the first minor pedicle. The preservation of the first minor pedicle in the split SCM flap in which the clavicular head was harvested could provide a reliable flap. The use of the sternal head till the occipital artery, leaving the minor pedicle and the clavicular head in situ, could enable safe reconstructions with the SCM flap. PMID- 15750416 TI - Effect of induction chemotherapy on changes of laminin and syndecan expression in oral squamous cell carcinomas: a prospective, randomized, clinicopathologic and immunohistochemical study. AB - Sixty patients with tumors of the floor of the mouth or of the tongue (T2N0-1 2M0) were randomized into three treatment groups. The first two groups participated in low-dose inductive chemotherapy, surgery, and then radiotherapy, whereas the third control group underwent only surgery and radiotherapy. In all three groups, studies were made of the stage, grade, sex, localization, extents of expression of the pretreatment laminin and syndecan-1 and the cancer specific survival rate, and the correlations among these. The response to neoadjuvant chemotherapy was assessed by means of a method that we developed, involving measurement of the degree of histologic regression observed in response to chemotherapy. Immunohistochemical methods were applied to investigate the changes in degree of expression of laminin and syndecan-1 in response to the medication and their correlations with the survival. As concerns the overall tumorfree survival rate, a significant difference was not found between the two chemotherapeutic groups. However, there was a significant difference between the survival indices of those who participated in cytostatic treatment (70%) and the control group (40%). In the clinical and immunohistochemical examinations, the initial laminin and syndecan-1 levels obtained from biopsy samples could be used as prognostic factors. Our model measuring the extent of histologic regression clearly demonstrated that the survival indices of the patients who responded to the neoadjuvant cytostatic treatment with adequate tissue regression were better than those of the patients who responded to the treatment to only a decreased extent or not at all. The changes in the expressions of laminin and syndecan-1 in response to cystostatic treatment proved to be important predictive factors. The increase or stagnation of these clearly forecast a good prognosis, whereas their decrease was a definite indication of poor prognosis. PMID- 15750417 TI - Effect of calcium sulfate-chitosan composite: pellet on bone formation in bone defect. AB - The purpose of this experiment was to study the effects of chitosan, calcium sulfate, and calcium sulfate-chitosan composite pellet on the osteogenesis of defective tibia in rabbits. Eighty New Zealand white rabbits, each weighing approximately 3 to 3.5 kg, were used for this study. A 1-cm ostectomy was made on the middle of the tibia of each rabbit with the periosteum preserved. Nothing was implanted in the control group (group 1), and five chitosan pellets (60 mg/pellet) were implanted in group 1, three OsteoSet pellets (100 mg/pellet) in group 3, and four calcium sulfate-chitosan composite pellets (1 pellet, 80 mg; calcium sulfate 40 mg/pellet, chitosan 40 mg/pellet) in group 4. For each group, a radiographic study, bone mineral density test, three-point bending test, and histologic examination were performed in the second, fourth, and sixth weeks. In the radiologic study, in group 1, cortical bone was not formed even at 6 weeks. In group 2, it was observed at 6 weeks. In groups 3 and 4, cortical bone was partially seen around the fourth week. At 6 weeks, it was clearly observed on both sides, and the projection of the marrow cavity became distinctive, so bone consolidation was considered to be much progressed. The bone mineral density test and three-point bending test results appeared to be highly similar in groups 3 and 4 and in groups 2 and 1. Particularly at 6 weeks, the measures for groups 3 and 4 were statistically significant compared with those for groups 1 and 2 (P < 0.05). In histologic examination, new bone formation began to be seen at 2 weeks in all groups, but it was more active and faster in groups 3 and 4. At 6 weeks, fibrous connective tissue still remained at the center in groups 1 and 2; however, the fibrous connective tissue at the center was replaced with callus, the bony bridge was obvious, and lamellation of callus was observed more in groups 3 and 4. The results indicate that chitosan pellets, OsteoSet, and chitosan-calcium sulfate composite pellets facilitate new bone formation on defected bone, and that particularly OsteoSet and chitosan-calcium sulfate composite pellets are more effective than chitosan. PMID- 15750419 TI - Antibiotic elution from hydroxyapatite cement cranioplasty materials. AB - Hydroxyapatite cements (HAC) are a contemporary material used for multiple cranioplasty applications. In an effort to decrease the risk of postoperative infection, mixing antibiotics into the material during intraoperative application is frequently done. It has been assumed, but never substantiated, that significant antibiotic release from the material occurs after implantation. Using standardized morphologies, a mixture of a specific HAC (Mimix bone void filler) and tobramycin antibiotic was prepared, hydrated in phosphate-buffered saline, and tested in vitro for as long as 22 days after preparation. The results show that the majority of the antibiotic (91%) was released within the first 24 hours, with the balance being eluted during the next 8 days. Overall, the release of tobramycin from Mimix bone void filler appears to fit the pattern of antibiotic release demonstrated to occur from other bioabsorbable ceramic-type carriers. PMID- 15750420 TI - Determinants of successful treatment of bimaxillary protrusion: orthodontic treatment versus anterior segmental osteotomy. AB - The purpose of this study was to investigate the differences in initial skeletal, dental, and soft tissue characteristics of bimaxillary protrusion (BP) patients to determine poor or good results with orthodontic treatment (OT) or anterior segmental osteotomy (ASO) with extraction of four first premolars. Lateral cephalometric radiographs of 46 adult Korean females with BP were analyzed before treatment (T0) and after treatment (T1). According to the measurements at T1, patients were classified into group 1 (poor result with OT, n = 12), group 2 (good result with OT, n = 11), group 3 (poor result with ASO, n = 5), and group 4 (good result with ASO, n = 18). Sagittal, vertical, dental, and soft tissue variables were measured. The differences at T0 among the four groups were compared by one way analysis of variance test and verified by Scheffe's multiple comparison test. Stepwise discriminant analysis was performed to find decisive predictors. Skeletal class II malocclusion tendency, less developed chin, and vertical facial growth pattern were related with group 1. Overly uprighted and less protrusive upper and lower incisor, near normal interincisal angle (IIA), less protrusive upper lip, and more obtuse lower nasolabial angle (NLA) were related with group 3. IIA, U1-NA distance, combination factor, interlabial gap, lower NLA, pterygomaxillary fissure-N, and posterior nasal spine-anterior nasal spine were selected as significant variables for discriminating the four groups. The percentage of correctly classified cases was 91.3%. In particular, the discriminant function showed the highest accuracy in the prediction of group 4. These variables and discriminant functions contributed to the differential diagnosis on BP to make a procedural decision between OT and ASO. PMID- 15750421 TI - Unilateral mandibular ramus elongation by intraoral distraction osteogenesis. AB - Successful correction of facial asymmetry by mandibular distraction osteogenesis relies on mastering vector control. Lack of necessary vector control continues to compromise the treatment modality. The purpose of this study was to describe a new simplified method for vector transfer and to evaluate this method according to the outcome and the efficacy of distraction osteogenesis in the correction of unilateral mandibular hypoplasia and asymmetry. Twenty-seven patients with unilateral hypoplasia of the mandibular ramus underwent unilateral mandibular distraction osteogenesis with intraoral distraction devices. Posterior-anterior and lateral cephalograms were analyzed by digitalization. Changes in sagittal, vertical, and transversal linear and angular dimensions after the distraction treatment were evaluated by measurements performed on headfilms taken before and after surgery. Means and variances were calculated for selected cephalometric variables for each time point. The differences between the control and the treatment side were calculated, statistically described, and compared with a paired Student t test. Correction of the mandibular asymmetry, chin position, and the canting of the occlusal plane was obtained in all patients clinically, as well as radiographically, by the use of intraoral unidirectional distraction osteogenesis. PMID- 15750422 TI - The central lip flap and nasal mucosal rotation advancement: important aspects of composite correction of the bilateral cleft lip nose deformity. AB - The columella, nasal tip, lip relationship in the secondary bilateral cleft deformity remains an enigma and a great challenge for the cleft surgeon. A subset of patients with bilateral cleft lip still require columellar lengthening and nasal correction, despite the advances in preoperative orthopedics and primary nasal corrections. An approach to correct this deformity is described. This consists of (1) lengthening the columella by a central lip advancement flap; (2) open rhinoplasty, allowing definitive repositioning of lower lateral cartilages, ear cartilage grafting to the tip and columella when necessary; (3) nasal mucosal advancement; (4) alar base narrowing; and (5) reconstruction of the orbicularis oris as required. Depending on the individual assessment of the patients, some of these steps were not performed, leaving the nasal mucosal advancement the most important aspect of the reconstruction. In a consecutive series of 72 patients with repaired bilateral cleft lip and palate, 17 patients have been treated with nasal mucosal rotation advancement and followed up for a maximum period of 10 years. With the use of this technique, the secondary bilateral cleft lip nose deformity has been successfully corrected. PMID- 15750423 TI - Natal teeth: a potential impediment to nasoalveolar molding in infants with cleft lip and palate. AB - The purpose of this article is to describe two cases of bilateral cleft lip and palate with natal/neonatal teeth. Clinical features, prevalence, possible complications, and treatment modalities are discussed. Two patients with cleft lip and palate and natal/neonatal teeth are described. The first patient was a 4 week-old infant with bilateral cleft lip and palate. On initial inspection an odontogenic vestige was noticed on the right lateral border of the premaxillary segment, bordering the cleft. The second patient was a newborn with a vestige on the right side of the premaxilla. At 20 days, another swelling representing a neonatal tooth on the left side was found. Because the teeth interfered with the fabrication and application of the nasoalveolar molding (NAM) appliance, they were removed from both patients. In the first patient, at 1 week after extraction, the NAM device was placed without difficulty. At 8 months, the infant had adapted well to the NAM device and nursed without problems. The second patient did not follow-up for the placement of the NAM device. In patients with cleft lip and palate with natal/neonatal teeth who require NAM, the tooth must be removed to facilitate the fabrication and placement of the device. Natal/neonatal teeth must be extracted with caution because the tooth buds of neighboring teeth may be damaged and remnants of the dental papillae may be left behind. Although general anesthesia is not always indicated for the removal of these teeth, in cases in which the premaxilla is loose, such as the current cases, general anesthesia is warranted. PMID- 15750424 TI - Video-assisted endoscopic transtemporal multilayer upper midface lift (MUM-Lift). AB - Minimally invasive surgery has been used successfully recently to improve brow ptosis and forehead rhytids. Because the face tends to age more vertically, rather than obliquely, it makes sense to perform the correction in a more vertical direction. Therefore, the authors introduce a video-assisted endoscopic transtemporal approach to allow a multiplanar (subperiosteal, sub-SMAS, and subcutaneous) vertical upper midface elevation (MUM-Lift) avoiding extensive lateral temporal and preauricular incisions. Between 1996 and 2003, 53 patients (8 males, 45 females; age, 47 +/- 6 years) underwent MUM-Lift. This includes simultaneous forehead lift, blepharoplasties and video-assisted transtemporal sub SMAS and subcutaneous upper midface tissue release, vertico-lateral repositioning, and fixation. Using the various planes of release in the midface produces less tension on each layer and provides a natural and improved facial rejuvenation of the forehead, as well as of the upper midface. The results confirmed that limited incision forehead-plasty techniques in combination with a transtemporal sub-SMAS and subcutaneous upper midface lift (MUM-Lift) reduces unnecessary vascular compromise on any particular layer and can be safely and predictably performed over the zygomatic muscle. This newly introduced procedure is an exciting feature that has evolved into a useful modality. It provides natural and improved facial rejuvenation. PMID- 15750425 TI - Post-traumatic severe trismus caused by impairment of the masticatory muscle. AB - Restriction of the mouth opening from a pathologic condition outside the temporomandibular joint is called a pseudo- or extra-articular ankylosis. The authors report two cases of severe post-traumatic pseudoankylosis. One case showed fibrous degeneration of the bilateral masseter muscles without a facial bone fracture, which caused severe trismus, a mouth opening of less than 2 mm, and gradually appeared after blunt injuries to the face. The other was a rare case accompanied with the bone formation in the masseter muscle and was diagnosed as myositis ossificans traumatica, which also presented as severe trismus, with a maximal mouth opening of 5 mm after facial violence. Both were surgically treated with dissection of the affected muscles. In addition, a hemicoronoidotomy was performed in the case of myositis ossificans traumatica. Although a conservative therapy with physical rehabilitation is the basic policy for the management of pseudoankylosis of the temporomandibular joint, a surgical treatment should be considered when the origin of the problems is an osteogenic character or severe extra-articular ankylosis resistant to conservative therapy before completion of true temporomandibular joint ankylosis. PMID- 15750426 TI - Evaluation of the adjunctive benefits of platelet-rich plasma in subantral sinus augmentation. AB - Subantral sinus augmentation is often necessary to permit placement of endosseous implants. Recent efforts to improve wound healing have focused on autogenous sources of bioactive mediators, such as platelet-rich plasma (PRP), which offer the potential to enhance the biological activity of bone replacement grafts. The purpose of this randomized, single-blinded, controlled study was to compare bone formation after subantral maxillary sinus augmentation with freeze-dried bone allograft (FDBA) plus PRP versus FDBA plus resorbable membrane. Ten patients underwent bilateral maxillary subantral sinus augmentation, with sites within subjects randomized to receive FDBA plus PRP or FDBA plus membrane. Core biopsy specimens were obtained 4.5 to 6 months after the grafting procedure at time of implant placement. Histomorphometric analysis revealed a significantly higher percentage of vital tissue in sinuses after treatment with FDBA and PRP (78.8 +/- 8.3) than with FDBA and membrane (63.0 +/- 15.7). Moreover, the percentage of bone formation in sinuses augmented with the combination of FDBA plus PRP (33.3 +/- 11.3) was nonsignificantly (P or =4) on day 0 (75 and 92%) and day 3 (88 and 100%) to predict severe NS were superior to the traditional clinical staging system on day 0 (63 and 58%). CONCLUSIONS: The TBAN score is an objective marker for predicting severe NS in children with TBM. PMID- 15750457 TI - Immunogenicity and reactogenicity of a combined adsorbed tetanus toxoid, low dose diphtheria toxoid, five component acellular pertussis and inactivated polio vaccine in six-year-old children. AB - BACKGROUND: The objective of this study was to assess the immunogenicity and reactogenicity of the combined adsorbed tetanus toxoid, low dose diphtheria toxoid, 5-component acellular pertussis and inactivated polio vaccine (TdcP-IPV) as compared with a pediatric dose diphtheria formulation, combined with adsorbed tetanus toxoid and 3-component acellular pertussis (DTacP), in 6-year-old children who were immunized with 4 doses of diphtheria-tetanus-whole cell cellular pertussis (DTwcP) plus oral polio vaccine (OPV) before 2 years of age, according to the local Spanish vaccination calendar. METHODS: One hundred ninety four healthy 6-year-old children were randomized to receive 1 dose of TdcP-IPV or 1 dose of DTacP and OPV. RESULTS: One month postvaccination, antidiphtheria and antitetanus titers were > or =0.1 IU/mL in 100% of patients in both study groups. TdcP-IPV reached 100% seroprotection rates against polio types 1, 2 and 3. In OPV recipients, these rates were 100, 100 and 96.8%, respectively. Seropositivity rates for pertussis toxin, filamentous hemagglutinin, pertactin and fimbrial components of the TdcP-IPV vaccine were 97.9, 89.6, 90.6 and 100%. The incidence of local and systemic reactions was 50.5 and 39.2% in the TdcP-IPV group and 59.4 and 38.5% in the DTacP plus OPV group, and no serious adverse events were reported. CONCLUSIONS: TdcP-IPV vaccine was shown to be immunogenic and safe when given as a booster in children 6 years of age who were primed with 4 doses of DTwcP and OPV. PMID- 15750456 TI - Comparative long term immunogenicity of two recombinant hepatitis B vaccines and the effect of a booster dose given after five years in a low endemicity country. AB - BACKGROUND: Few data are available concerning the long term immunogenicity of the pediatric doses of hepatitis B vaccines given to preteenagers. The long term effect of the booster dose in teenagers is unknown. We evaluated the immunogenicity of 2 pediatric hepatitis vaccines after primary vaccination and after a booster dose. METHODS: A prospective 15-year follow-up study of the immunogenicity of 2 hepatitis B vaccines was initiated in 1995 in Quebec City, Canada. One year apart, 1129 children 8-10 years old received Engerix-B 10 microg (EB), and 1126 received Recombivax-HB 2.5 microg (RB) vaccine after a 0-, 1-, 6 month schedule. After 5 years, one-third of the 2 cohorts were randomly selected. A booster dose of EB 10 microg or RB 5 microg was administered according to the vaccine used in the primary immunization. Antibodies were measured before, 1 month after and 1 year after the booster injection. RESULTS: Before the booster dose, anti-HB surface antibody (HBs) was detected in 94.7% of the EB subjects and in 95.2% of the RB subjects (P = 0.85). The geometric mean titer (GMT) was higher in the EB than in the RB group (252 mIU/mL versus 66 mIU/mL, P < 0.0001). One month after the booster, 99.7% of subjects in the EB group and 99.6% in the RB group had a detectable anti-HBs, and 99.0 and 99.3%, respectively, had anti-HBs > or =10 mIU/mL. The anti-HBs GMT was 113,201 mIU/mL in the EB and 16,623 mIU/mL in the RB groups (P < 0.0001). One year after the booster, 99.3% of subjects in the EB group and 100% in the RB group had detectable anti-HBs, and 97.9 and 98.5% respectively, had anti-HBs > or =10 mIU/mL. The anti-HBs GMT was 14,028 mIU/mL in the EB and 3437 mIU/mL in the RB group (P < 0.0001). CONCLUSIONS: The immunity persists for at least 5 years after the primary vaccination with both pediatric vaccines in 99% of children vaccinated at the age of 8-10 years. It confirms that no booster is needed at that point. PMID- 15750458 TI - Oral oseltamivir improves pulmonary function and reduces exacerbation frequency for influenza-infected children with asthma. AB - BACKGROUND: Among asthmatic children, influenza is associated with increased hospitalizations. Although vaccination is safe and effective among asthmatic children, its protective efficacy varies and uptake rates can be low. In comparison, oseltamivir (Tamiflu) is effective against all influenza strains and can reduce the severity and duration of influenza among adults and children. This study determined the effects of oseltamivir among influenza-infected children with asthma. METHODS: Asthmatic children (6-12 years of age) were randomized to receive oseltamivir (2 mg/kg) or placebo twice daily, as a syrup. The primary efficacy endpoint was the time to freedom from illness. Secondary endpoints included the area under the symptom score-hour curve, the proportion of patients with asthma exacerbations and changes in forced expiratory volume at 1 second during the dosing period. Analysis was performed for both the intent-to-treat infected (n = 179) and per protocol (n = 162) populations. RESULTS: The primary endpoint for this study was not met. Oseltamivir tended to reduce the time to freedom from illness in the intent-to-treat infected population (10.4 hours, 8%; P = 0.5420), the per protocol population (24.3 hours, 17%; P = 0.1607) and patients who started treatment <24 hours after symptom onset (39.8 hours, 25%; P = 0.0780). However, an improvement in pulmonary function was observed. The improvement in forced expiratory volume at 1 second was significantly greater among oseltamivir-treated patients (10.8% versus 4.7%; P = 0.0148). Oseltamivir treated patients also experienced fewer asthma exacerbations up to day 7 (68% versus 51%; P = 0.031). Oseltamivir was safe and well-tolerated. CONCLUSIONS: Oseltamivir is safe and well-tolerated among asthmatic children, may reduce symptom duration and helps improve lung function and reduce asthma exacerbations during influenza infection. PMID- 15750459 TI - Vaccine effectiveness and severity of varicella among previously vaccinated children during outbreaks in day-care centers with low vaccination coverage. AB - BACKGROUND: Varicella vaccine effectiveness (VE) during outbreaks has been reported to be 71-100% against any disease and >90% against moderate/severe disease even in day-care centers (DCCs) and schools with low vaccination rates. A recent report suggested an effectiveness rate of 44% during a DCC outbreak despite a high vaccination rate. AIMS: To reassess vaccination coverage, VE and severity of disease among previously vaccinated children after exposure during DCC outbreaks in northern Israel, where vaccination rates are low. METHODS: During January to June 2003, active surveillance for varicella among children in northern Israel revealed outbreaks in 8 DCCs with children 3-6 years of age. Data concerning symptoms of the disease and the age at vaccination (for previously vaccinated children) were obtained from parents and health care providers for children who contracted the disease. Analysis of VE was limited to children who were continuously enrolled in DCCs during the outbreaks. RESULTS: The overall vaccination rate was 37%. The incidences of natural varicella and breakthrough varicella (BV) were 79 of 153 [52%; 95% confidence interval (CI) 44-60%] and 37 of 89 (41.5%; 95% CI 31-52%), respectively. VE was 20% (95% CI 0-40%) against disease of any severity and 93.4% (95% CI 75-98%) against moderate/severe disease. Ninety-four percent and 14% of children with BV and natural varicella, respectively, had mild disease (P < 0.001). The odds ratio for BV was 17 (95% CI 2.18-118) for children vaccinated >2 years before the outbreak. CONCLUSIONS: During varicella outbreaks in DCCs with low vaccine coverage, previous vaccination provided poor protection against chickenpox, mostly among children who had been vaccinated >2 years earlier, but the disease appeared to be much milder among children with BV than among nonvaccinated children. PMID- 15750460 TI - Malignancy in perinatally human immunodeficiency virus-infected children in the United States. AB - OBJECTIVES: To determine the incidence of and factors associated with malignancy in perinatally human immunodeficiency virus (HIV)-infected children in the United States. METHODS: Included were 2969 children followed in the Pediatric AIDS Clinical Trials Group (PACTG) 219/219C cohort from 1993 through 2003. Cancer incidence by sex, race, age, histology and highly active antiretroviral therapy (HAART) era (pre-HAART, 1993-1997; HAART, 1998-2003) was estimated, and the standardized incidence ratio contrasting infected and uninfected children was determined. Poisson regression was used to further investigate the relation between HAART use (> or =3 drugs of > or =2 classes, 1 of which was a protease inhibitor), CD4% and cancer. RESULTS: There were 37 cancers (17 prevalent and 20 incident) diagnosed in 2969 children for a prevalence of 0.6% [95% confidence interval (CI), 0.3, 0.9] and an incidence of 1.56/1000 person-years (95% CI 0.95, 2.41). Compared with uninfected children, the standardized incidence ratio was 10.08 (95% CI 5.87, 16.14). Incidence did not significantly differ by sex, race, age or HAART era. Of the cases, 35% were immunocompetent (CD4 > or =25%), 25% were moderately immunosuppressed (15%< or = CD4 < or =24%) and 40% were severely immunosuppressed (CD4 <15%) at diagnosis. In multivariate regression, the cancer rate was 3.09 (95% CI 1.22, 7.85) times higher in children with < or =2 years of HAART use than in children with >2 years of HAART and 3.20 (95% CI 1.32, 7.76) times higher in children with CD4 <15% at cohort enrollment than in children with CD4 > or =15%. CONCLUSION: Cancer incidence in this U.S. pediatric cohort was lower than that of European cohorts but was markedly higher than that of HIV uninfected children. Cancer incidence was highest in children who were severely immunosuppressed and in children who received HAART for < or =2 years. PMID- 15750461 TI - Effect of the seven-valent conjugate pneumococcal vaccine on carriage and drug resistance of Streptococcus pneumoniae in healthy children attending day-care centers in Lisbon. AB - AIMS: Prospective study to evaluate the impact of the 7-valent pneumococcal conjugate vaccine (Prevenar) on the nasopharyngeal (NP) carriage of drug resistant Streptococcus pneumoniae (DRPn), by healthy children attending day-care centers (ages 6 months-6 years). METHODS: Vaccinees (238 children) who received vaccine and controls (457 children) were followed for carriage of total S. pneumoniae and DRPn and for the serotypes and genetic backgrounds of DRPn during 6 consecutive sampling periods between May 2001 and February 2003. RESULTS: We detected no significant differences between vaccinees and the control group in the total carriage rate of Pn (average, 68%) or in the frequency of carriage of DRPn (average, 38%), including the frequency of penicillin-nonsusceptible strains (average, 24%). In contrast, there was a decline in the carriage of DRPn with vaccine serotypes which was compensated by the appearance and gradual increase in the frequency of DRPn expressing unusual serotypes (6A, 10A, 15A and 15C, 19A, 23A, 33F) which were not present in the vaccine as well as an increase in nontypable strains. The majority of the DRPn with unusual serotypes showed different pulsed field gel electrophoresis patterns indicating replacement of the original resistant flora by other clonal types of drug-resistant bacteria. Antibiotic consumption and the frequency of respiratory tract infections were similar among the vaccinees and controls. CONCLUSIONS: Pneumococcal vaccination did not change the frequency of carriage of drug-resistant strains being the initially dominant vaccine serotypes replaced by others expressing nonvaccine serotypes. Reduction in the carriage of DRPn may require a combination of the conjugate vaccine and a decrease in antibiotic pressure. PMID- 15750462 TI - Oral fluid human immunodeficiency virus tests: improved access to diagnosis for infants in poorly resourced prevention of mother to child transmission programs. AB - BACKGROUND: Perinatal exposure of infants in low resource settings generates the bulk of pediatric human immunodeficiency virus (HIV) disease globally. The HIV status of these infants is established by testing serum for anti-HIV antibodies at 12 months of age in Prevention of Mother to Child Transmission (PMTCT) programs because polymerase chain reaction testing is unavailable. The diagnostic accuracy of 2 oral fluid (OF) HIV tests has not been previously evaluated in children. METHODS: A serum and 2 OF HIV tests were performed at 12 months of age in a cohort of 321 vertically exposed children in a prospective, longitudinal study at a secondary level hospital in Johannesburg, South Africa during a 14 month period preceding October 2003. The 3 HIV tests were performed independently of each other by personnel blinded to the child's true HIV infection status, the reference standard used for comparison. RESULTS: HIV testing was performed at a median age of 12.1 months. The true HIV infection status of 310 of 321 (97%) children was determined. In comparison with serum testing results, OF HIV tests reduced the percentage of children requiring repeat HIV tests from 45% to 8-12%. The abilities of OF and serum to predict an HIV-uninfected status were comparable with negative predictive values >99%. Interpretation of HIV tests in conjunction with simple clinical assessment further improved the predictive value of the test. CONCLUSIONS: OF HIV tests perform well in children and have the potential to increase accessibility and acceptability of HIV diagnosis for infants in the context of PMTCT programs in low resource settings. PMID- 15750463 TI - Peak rotavirus activity shifted from winter to early spring in Japan. AB - BACKGROUND: Since 1910, there have been many studies on acute gastroenteritis in children in Japan. These diseases, namely Kasei-shoni-kolera (pseudocholera infantum) or banshu-otosho (late autumn vomiting disease), are historically known to occur in the cooler season with a peak in November or December. Earlier we confirmed their causation by rotaviruses but found peaks in January or February from 1974 to 1981. The aim of the present study was to confirm the temporal shift in peak rotavirus activity. METHODS: Under the National Epidemiological Surveillance of Infectious Diseases program from 1983 through 2003, rotavirus positive patients 0-3 years old and clinically diagnosed with "infantile vomiting and diarrhea" at sentinel clinics were examined. Fecal samples were screened by electron microscopy and/or using commercial latex agglutination kits at prefectural/municipal Public Health Institutes, and we determined the trend for the "peak" month during 21 seasons. RESULTS: Peak rotavirus activity shifted gradually from January to March during the 21 consecutive seasons. The mean duration from December to the peak month (mean beginning peak duration) of the rotavirus season significantly varied among 3 periods of 7 consecutive seasons (1.7 +/- 0.5 months in 1982/1983-1988/1989, 2.3+/-0.8 months in 1989/1980 1995/1996, and 3.1 +/- 0.7 months in 1996/1997-2002/2003, respectively; P = 0.0026 by 1-way analysis of variance). This time series shift in the peak rotavirus infection was statistically significant (P = 0.0003 for trend). CONCLUSION: Our findings confirmed that the temporal trend in peak rotavirus activity in Japan has shifted gradually from winter to early spring for unknown reasons. PMID- 15750464 TI - Severe chronic active Epstein-Barr virus infection mimicking steroid-dependent inflammatory bowel disease. AB - Severe chronic active Epstein-Barr virus infection is a rare, debilitating, nonneoplastic, inflammatory disorder for which there is no definitive treatment. We describe a Caucasian child who initially presented with clinical and histopathologic features consistent with inflammatory bowel disease, for whom additional investigations led to the diagnosis of severe chronic active Epstein Barr virus infection. PMID- 15750465 TI - Evaluation of recurrent pneumonia. PMID- 15750466 TI - Probiotics and the treatment of infectious diarrhea. PMID- 15750467 TI - Recurrent papular urticaria after varicella immunization in a fifteen-month-old girl. AB - Varicella-like rash after immunization with the live attenuated varicella vaccine is relatively common. Such vaccine-associated rashes generally consist of fewer lesions than occur in chickenpox. We describe a 15-month-old girl who experienced the onset of recurring papular urticaria after varicella immunization. The rash was varicella-like and thought by us to be caused by vaccine virus. PMID- 15750468 TI - Salmonella typhi-infected lung hydatid cyst. AB - A 12-year-old girl presented with hemoptysis, fever and weight loss. She was initially diagnosed and treated for presumed tuberculosis and a lung abscess. Salmonella typhi was isolated from the abscess. Hydatid cyst was diagnosed at surgery. This is the first reported case of a pulmonary hydatid cyst infected with S. typhi. PMID- 15750469 TI - Persistent parvovirus-associated chronic fatigue treated with high dose intravenous immunoglobulin. AB - We report a 16-year-old boy with no evidence of immunodeficiency who had a 2-year history of chronic fatigue, low grade fever and slapped-cheek rash associated with chronic parvovirus B19 viremia. Prolonged intravenous immunoglobulin therapy resulted in resolution of his symptoms and viremia. Intravenous immunoglobulin may be useful in the resolution of parvovirus viremia regardless of immune status. PMID- 15750470 TI - Emergence of community-acquired methicillin-resistant Staphylococcus aureus in complicated parapneumonic effusions. AB - A retrospective study was conducted of all patients with complicated parapneumonic effusion discharged between January 1, 1994 and March 31, 2004. We identified 54 patients. The number of patients with complicated parapneumonic effusion per 10,000 admissions increased from 4.4 in 1994 to 18.4 in 2000 (P = 0.03) and then decreased to 13.9 in 2003 (not statistically significant). Community-acquired methicillin-resistant Staphylococcus aureus accounted for none of the 36 cases from 1994 through 2001 and 4 of the 18 cases from 2002 through 2004. PMID- 15750471 TI - Nasal carriage of methicillin-resistant Staphylococcus aureus in school children without identifiable risk factors in northern taiwan. AB - In a nasal culture survey of school children, 5 (1.9%) of 262 school children and 18 (13%) of 137 health care workers (HCWs) were colonized with methicillin resistant Staphylococcus aureus (MRSA). One common genotype accounted for most isolates from the school children and from the HCWs. Based on genotyping, MRSA strains circulating in the community in Taiwan were similar to the hospital strains. PMID- 15750472 TI - Lactobacillus rhamnosus GG bacteremia associated with probiotic use in a child with short gut syndrome. AB - Probiotic agents are increasingly used for the treatment and prevention of a variety of infectious and inflammatory conditions. They are generally safe, but complications of probiotic use can occur. In this report, we describe bacteremia after ingestion of a Lactobacillus rhamnosus GG probiotic tablet in a child with short gut syndrome. We used sequencing of the ribosomal operon region and strain typing with pulsed field electrophoresis of the isolates to show identity between the tablet and bloodstream isolates. PMID- 15750473 TI - Eighteen-month-old girl with respiratory syncytial virus infection and diffuse nodular lung infiltrates. PMID- 15750474 TI - Atypical presentation of parvovirus infection. PMID- 15750475 TI - Staphylococcus aureus infection caused by a Panton Valentine leukocidin-producing strain. PMID- 15750476 TI - Intracranial hydatid cyst: can we prevent misdiagnosis? PMID- 15750477 TI - Serologic diagnosis of congenital rubella: a cautionary tale. PMID- 15750478 TI - Use of 16S ribosomal DNA polymerase chain reaction to identify Haemophilus influenzae type b as the etiology of pericarditis in an infant. PMID- 15750479 TI - Henoch Schonlein purpura after varicella infection. PMID- 15750480 TI - NMC is to set standards for advanced nursing practice. PMID- 15750481 TI - The long-awaited Mental Capacity Bill has arrived. PMID- 15750482 TI - Assessing community nurses' level of knowledge of lymphoedema. AB - As part of a major project to develop a primary care trust-based framework of lymphoedema management, the educational needs of community nurses regarding the care of patients with lymphoedema were assessed using focus groups and questionnaires. Community nurses assessed their current knowledge and skill in the care and management of patients with lymphoedema as adequate or poor. They were concerned about their lack of knowledge and skill and were uncertain regarding their role relative to other professions involved in the care of this patient group. At the same time they understood the importance of their role in providing ongoing care, recognizing problems, offering sound advice and referring on to a specialist practitioner when necessary. Any education provision that prepares community nurses for their role within a framework of lymphoedema management should emphasize the important place they occupy in providing long term care for this patient group. PMID- 15750483 TI - Chronic obstructive pulmonary disease: non-pharmacological approaches. AB - Chronic obstructive pulmonary disease (COPD) is a progressive, smoking-related, chronic disease that eventually impacts on every area of the lives of the patient and his/her family and carers. The nature of the disease - chronic, irreversible airflow obstruction - limits the impact of drug therapy, particularly as the disease progresses. Increasing breathlessness on exertion saps confidence, reduces self-esteem and produces dependence. In addition, frequent exacerbations utilize health service resources. Non-pharmacological approaches, such as pulmonary rehabilitation, together with lifestyle advice that enables patients to help themselves manage exacerbations, maintain independence and quality of life and make the most of life with COPD can benefit both patients and the health service. PMID- 15750484 TI - Nursing role in the multidisciplinary management of motor neurone disease. AB - Motor neurone disease (MND) is a debilitating and progressive neurological disorder in which degeneration of the motor neurones results in muscle weakness and wasting. Recent high-profile media coverage about assisted suicide and euthanasia has increased awareness of MND and yet still relatively little is known or understood about it, even among the healthcare profession. This article looks at the key areas in management of patients with MND, including the role of the nurse and the importance of a well-coordinated multidisciplinary team. Ways to deal with communication problems, respiratory dysfunction, feeding and nutrition, and legal and ethical issues are examined. Good management of patients with MND involves a hospital and community-based multidisciplinary team. The essence of care is good symptomatic management of respiratory failure and nutritional problems that result from difficulties in swallowing and increasing dependency. Palliative care should be introduced before the terminal stages after careful discussion with the patient and carers. Most patients with MND die from pulmonary infections or respiratory failure, but contrary to popular belief, death by choking attacks is rare and the final stages of MND are usually peaceful and dignified. PMID- 15750485 TI - The role of the expert patient in the management of chronic illness. AB - The concept of the expert patient is an increasingly important one in healthcare policy and delivery. To date, however, there has been relatively limited consideration of the nature and characteristics of the knowledge and skills underpinning the role of expert patient or the relationship to more traditional health professional expertise. This article considers the emerging concept of the expert patient in the UK, its relationship to the emergence of the expert patient elsewhere (notably in Canada), together with exploration of the supporting rationales, assumptions and possible implications associated with the initiative. PMID- 15750486 TI - Clinical skills: how to remove and change a suprapubic cathether. AB - Suprapubic catheterization is becoming more widely used to drain the urinary bladder either short or long term or is inserted where initial urethral or recatheterization is problematic. No matter where you work as a nurse, either in the hospital, community, hospice or nursing care home, sooner or later you will have to deal with a patient with a suprapubic catheter in situ. One of the concerns nurses encounter when changing the suprapubic catheter is what action to take when the catheter appears to have become stuck when removing it. The main cause of this problem is owing to a 'cuffing' effect occurring to the deflated catheter balloon, especially if using 100% silicone catheters. This article looks at the problem of removing a suprapubic catheter that has become stuck and how to change such catheters safely. PMID- 15750487 TI - Examining adult male genitalia: providing a guide for the nurse. AB - This article provides the nurse with an overview and guidance concerning examination of the adult male genitalia. Issues such as ensuring privacy, developing a relationship and explaining the procedure are discussed. Advice is provided that will allow the nurse to perform the examination using sight, palpation and transillumination, activities that are central to the examination. An overview of the structure and function of male genitalia is provided. Examination of the male genitalia is an intimate activity and in this article emphasis is placed on the issues of informed consent and the importance of ensuring that each patient has a chaperone present during the examination. Detailed discussion is provided to enable the nurse to use a step-by-step approach to ensure a thorough and detailed examination is performed. A checklist and review of the examination process is included. PMID- 15750488 TI - General nurse who gave inaccurate information in healthcare records. PMID- 15750489 TI - Early postoperative feeding and abdominal gynaecological surgery. AB - Since the 1990s, the traditional postoperative feeding protocol has been challenged for patients who have undergone major abdominal gynaecological surgery, and value of extended delays in commencing oral intake has been questioned. This review aims to determine whether early rather than delayed postoperative feeding following major abdominal gynaecological surgery has an impact upon recovery as measured by length of hospital stay. The Cochrane Library, Embase, CINAHL and Medline were searched for randomized controlled trials (RCTs) which compared early postoperative oral intake with delayed (traditional) postoperative oral intake for patients who had undergone major abdominal gynaecological surgery. Three RCTs were identified and available for consideration. Although study quality was variable, they suggest that early postoperative feeding appears to be safe and well tolerated after most abdominal gynaecological surgery and leads to a considerable reduction in hospital stay of approximately one day. However, patients' preference and costs remain unclear. Therefore, further well-designed studies are needed to identify the comprehensive effectiveness of early postoperative feeding protocol. PMID- 15750490 TI - Law concerning organ transplants and dead donors in the UK. PMID- 15750491 TI - Nurses must strengthen the nurse/patient relationship. PMID- 15750492 TI - Drug fiasco highlights need for more openness. PMID- 15750493 TI - New opportunities for district nursing: chronic disease and matrons. PMID- 15750494 TI - Identification of skin cancers 1: benign and premalignant lesions. AB - Skin cancers are the most commonly diagnosed cancer in the UK, and their incidence is growing. They can present in all age groups, and are particularly common in older people, but the incidence among younger people is increasing. However, the majority of skin lesions are not malignant. In this, the first of two articles, the author discusses the range of common benign growths that can be mistaken for cancers, as well as the precancerous lesions that can become a cause for concern if untreated. The second article will examine the common malignant lesions. PMID- 15750495 TI - Outpatient stem cell transplant: effectiveness and implications. AB - The aim of this mini-review (Griffiths, 2002) was to assess whether outpatient autologous haematopoietic stem cell transplant (AHSCT) is as effective as the traditional inpatient approach in terms of bed occupancy and morbidity. The Cochrane Library, Medline, Embase and CINAHL databases were searched for relevant studies. No systematic reviews or randomized control trials were found and after applying inclusion criteria, four non-randomized control trials were identified. The studies were conducted in North America, where high costs associated with AHSCT have led to the development of outpatient programmes designed specifically to reduce use of costly inpatient facilities. From the evidence available, it appears that outpatient AHSCT is not associated with increased morbidity and studies found either in favour of the intervention group or that there was no statistically significant difference between the two groups. The studies were unanimous in their findings that outpatient AHSCT is associated with a significantly reduced bed occupancy outcome. These findings may have a significant implication for the advancing practice and workload of community nurses. It is evident that good quality trials are needed in the UK to better and more relevantly address this question before implementing such programmes but it seems that outpatient AHSCT is achievable in carefully selected patients. PMID- 15750496 TI - Making the most of teams in the mentorship of students. AB - Community nurses are rightly expected to act as mentors. This role offers students valuable opportunities to gain insight into community care and contributes to the development of future services and the professional growth of individual mentors. There can be tension, however between the demand of nurses' clinical roles and the need to support students. This article identifies ways in which teams can get the most out of mentoring, focusing on the importance of strong clinical leadership and the need for nurses to "own" the mentorship role and to work with other stakeholders. Finally, examples of initiatives from one inner-London trust are offered to demonstrate robust ways of enabling practice based education. PMID- 15750497 TI - Stoma care accessories: an overview of a crowded market. AB - While many stomas do not develop any problems, up to 55% of them do. Stoma accessories are often used to either prevent stoma problems or resolve them, but there is a potentially baffling array of products available. This article provides an overview of the accessories currently available, to assist the community nurse in the care of patients with stomas. PMID- 15750498 TI - Taking the Queen's Nursing Institute into the future. Interview by Tom Pollard. PMID- 15750499 TI - Diabetes and peripheral neuropathy: keeping people on their own two feet. AB - Peripheral neuropathy is one of the most common forms of diabetes-related neuropathy, and most commonly affects the feet. Loss of sensation leaves the feet at risk of damage--from environmental hazards, incorrectly fitting footwear or un felt pressure damage--which can lead to the need for amputations. This article discusses how to reduce the risk of peripheral neuropathy through glycaemic control, and the steps that need to be taken to reduce the risk of injury on the neuropathic foot. PMID- 15750500 TI - The Mental Capacity Bill 2004: its impact on district nurse practice. AB - The government made clear its intention to reform the law relating to mentally incapable adults in the Queen's speech at the state opening of Parliament in November 2004. In this article Richard Griffith and Cassam Tengnah outline the main provisions of the Mental Capacity Bill and assess its likely impact on district nurses who care for adults with incapacity. PMID- 15750501 TI - Depression and COPD in older people: a review and discussion. AB - Depression affects approximately 40% of patients with chronic obstructive pulmonary disease (COPD) and is largely untreated. This review draws on the author's studies and the available literature to examine the problem. Older people with COPD experience moderate levels of depression, but this goes largely unrecognized and untreated. The prevalence, severity and risk factors for depression in this population are discussed. Treatment, in the form of pulmonary rehabilitation and antidepressant drug therapy, are discussed, but in the absence of well-controlled trials, it is difficult to draw any firm conclusions about the benefits of these therapies. However, an appropriate home care package, with the support of respiratory nurses, is of some benefit to housebound patients with COPD because it helps to relieve depression and improve their quality of life, especially among those with a high level of depressive symptoms. PMID- 15750502 TI - Improving chronic disease management. PMID- 15750503 TI - Nurses' retirement age may be increased from 60 to 65. PMID- 15750504 TI - Patient safety must be an absolute priority for the NHS. PMID- 15750505 TI - Preimplantation genetic diagnosis to prevent disorders in children. AB - Preimplantation genetic diagnosis (PGD) is an alternative reproductive option for couples at risk of having a child affected with a genetic disorder. Although prenatal diagnosis (PND) has been available for many years, it is not acceptable to many owing to issues relating to termination of pregnancy. PGD involves assisted-reproductive technology, even though most couples undertaking it are fertile. However, if the treatment is successful, the couple will not have to consider PND. PGD is only available at a small number of centres and for a limited number of genetic conditions. It is a complex and time-consuming procedure. The success rate is around 20%, consequently, there is a relatively low chance of success and this requires careful consideration by couples who generally can become spontaneously pregnant. However, PGD is now more widely understood and available to all at the point of referral. This article sets out to explain the PGD procedure, its implications, limitations, and regulation in the UK and to discuss associated dilemmas. PMID- 15750506 TI - Nurses' views of longitudinal genetic screening of and research on children. AB - There is a lack of empirical data exploring ethical issues of genetic screening and longitudinal research involving children. Therefore, this pilot interview study explored the perceptions of nurses and midwives in relation to their involvement in an ongoing genetic preventive screening process involving children - the All Babies in South-east Sweden (ABIS) study (n=17,005). Data were collected through semistructured interviews with 10 nurses involved in all information and sampling procedures. While providing the preliminary nature of this study, it supports the idea of the importance of further research, both from a nursing professional perspective and from other parties involved in clinical research. The findings made in this study suggest that for such studies it is vital that nurses and midwives are fully informed about aims, methods, and potential intervention/prevention since in many cases they have a central role in several areas of screening and clinical longitudinal research involving children, e.g. information to potential research participants, obtaining informed consent, and data collection. With a thorough understanding of the research, including both basic aims and methods as well as potential future prevention aims, the nursing staff involved will be better placed to help participants make an informed choice and to provide additional information to the participants. Further research may be needed that aims to develop effective methods in preparing data collectors. It is also suggested that the design of the information process, and especially in longitudinal research involving young children, is of utmost importance before such studies are commenced. PMID- 15750507 TI - Paediatric asthma: overcoming barriers to an improved quality of life. AB - Paediatric asthma has a considerable impact on both society, in terms of healthcare resources, and patients and their families, in terms of impaired quality of life. The principal goals of asthma treatment are to achieve and maintain control of symptoms. Achieving these goals may involve long-term use of appropriate medication in the form of an inhaled corticosteroid (ICS) and a long acting ss-agonist (LABA). However, many patients with paediatric asthma are not currently achieving symptom control. The main barriers to asthma control are underuse of effective therapies, inappropriate choice of drug delivery devices and a lack of patient or parent/guardian education regarding the disease and its treatment. By addressing and overcoming these barriers to asthma control, the quality of life of patients and their families may be significantly improved. PMID- 15750508 TI - Caring for catheterized patients: urinary catheter maintenance. AB - This article brings together the evidence and opinions surrounding the ongoing problem of catheter blockage and how to help maintain catheter life. The reasons behind blockage are examined as well as the roles that pH testing and building a comprehensive catheter history, including all aspects of patients' general health, play in caring for the catheterized patient. The use of catheter maintenance solutions is addressed with discussion on smaller volumes, the use of double instillations and gentle technique for a shorter time of instillation and improvement in catheter-intervention techniques. PMID- 15750509 TI - Nurse who used her registration for her own commercial interests. PMID- 15750510 TI - Exploring the conditions leading to stoma-forming surgery. AB - The most common disease leading to the formation of a stoma is cancer. However, there are many other diseases that affect the gastrointestinal or urinary system that may also require either a temporary or permanent stoma to be formed (a colostomy, ileostomy or urostomy). Stoma-forming surgery may be undertaken for a number of reasons, such as to eradicate a disease or improve the patient's quality of life. Cancer, inflammatory bowel disease and diverticular disease are the most common conditions that lead to stoma formation. However, faecal incontinence, familial adenomatous polyposis, Hirschsprung's disease, spina bifida and Ehlers-Danlos syndrome may also require a stoma to be formed. The nurse's role in caring for these patients at a potentially traumatic period of their life is discussed. PMID- 15750511 TI - Dual diagnosis: co-existence of drug, alcohol and mental health problems. AB - The Department of Health (DoH) published a set of good practice implementation guidelines on dual diagnosis in May 2002. This guidance suggests that in order to improve the prognosis for clients who have mental health problems and who drink or take drugs problematically, mental health and substance misuse services should adopt an integrated service model. There is a considerable amount of American based research supporting this approach, but little evidence from the UK researchers demonstrating its application in the UK. This article offers an example of a service that has been developed in the city of Nottingham and argues that this client group will be served most effectively if mental health services support specialist dual-diagnosis resources. Integrated care pathways for this client group can be developed and led by specialist clinicians acting as consultants to mental health services (DoH, 2002a). This consultancy role within mental health services enhances the links needed between substance misuse and mental health services. As a result, specialist dual-diagnosis teams are best placed to increase positive prognoses for clients by ensuring evidence-based substance misuse skills are utilized and adapted by mental health teams to ensure fully integrated care coordination. PMID- 15750512 TI - Removal, retention and storage of organs and tissue in the UK. PMID- 15750513 TI - Sustained silver-releasing dressing in the treatment of diabetic foot ulcers. AB - This study investigated the clinical performance and safety of a sustained silver releasing foam dressing, Contreet Foam, in the treatment of diabetic foot ulcers. Twenty-seven patients with diabetic foot ulcers of grade I or II (Wagner's classification) were followed for six weeks: one week run-in using Biatain dressings, four weeks' treatment with Contreet dressings. Four ulcers healed during the four-week treatment with Contreet 56% in average. Contreet Foam showed good exudate management properties and was considered easy to use. Only two infections occurred showed that all six of the non-study ulcers developed an infection during the study. All ulcers (study ulcers as well as non-study ulcers) were treated according to good practice of diabetic wound care. There were no directions for the treatment of secondary wounds. No device-related adverse events were observed. This study demonstrated that Contreet Foam is safe and easy to use and effectively supports healing and good wound progress of diabetic foot ulcers. PMID- 15750514 TI - Better communication with older people is needed. PMID- 15750516 TI - Survey of UK hospice and specialist palliative care adult bereavement services. AB - METHOD: Postal survey of UK hospices and specialist palliative care services providing adult bereavement support. ANALYSIS: Descriptive statistics and content analysis of free text replies. RESULTS: Three hundred services were identified, with 248 valid responses (83%). Of these, 198 (80%) were in England and 180 (73%) were associated with inpatient units. Most had been in existence for at least 10 years. Paid staff were used by 219 services (88%) and volunteers were involved in 168 services (68%). A small minority did not provide supervision for their bereavement staff. A quarter of services had insufficient staff. The most common activities were individual support, telephone support, written information, memorialization events and group support. Ninety-five services (43%) formally assessed the need for individual support. One hundred and fourteen services (51%) had no formal mechanisms for bereaved people to provide feedback about such support. Formal audit and evaluation of bereavement services was uncommon. CONCLUSIONS: The main elements of bereavement support can be identified but their combination varies. Assessment of people for individual support varies and the small size of many services may inhibit the effective delivery of support. Audit and evaluation of bereavement support may need to be developed. PMID- 15750517 TI - Time to debunk some palliative care myths. PMID- 15750518 TI - Changing practice: use of audit to change oral care practice. AB - Although oral problems are common in palliative care, oral care can be a neglected area of practice. This article discusses the introduction of an oral care protocol and standard in one palliative care unit. The project used audit tools developed by Lee et al (2001) and involved a survey of the oral care knowledge of nursing and medical staff, and a retrospective survey of current practice using 50 sets of patients' notes. A standard and documentation were developed with the aim of ensuring oral care was both consistent and in line with best practice. Training was provided to all staff within the unit. Three months later oral care practice was audited. The results suggested improvement in all aspects of oral care practice. Dissemination of oral care documentation and training across the trust is underway. This article describes the processes used, highlighting the importance of assessment in oral care and the need for teamwork in rolling out changes both within and beyond a single unit. PMID- 15750519 TI - Cardiopulmonary resuscitation policies in northwest England hospices: a telephone survey. AB - DESIGN: A survey into existence of resuscitation policies in hospices in Northwest England. METHODS: All 25 hospices in the region were contacted. The clinical services manager or equivalent person was interviewed by telephone by means of a structured questionnaire. RESULTS: The telephone survey had a 96% response rate. Sixteen (67%) hospices did not have a resuscitation policy although 50% of this group were developing a policy. Only eight (33%) hospices had a formal policy at the time of interview. Twenty hospices (83%) provided staff with annual training in basic resuscitation. One hospice (4%) discussed cardiopulmonary resuscitation (CPR) with all patients admitted, whereas six (25%) discussed CPR only if the patient raised the topic. Five hospices (21%) would advise the patients that they should be cared for in an acute hospital, as no resuscitation would be provided. Only four hospices (17%) had written information on resuscitation. DISCUSSION: There was significant variation in the production and adherence to guidelines on resuscitation, with some patients being denied access to specialist palliative care units, as they would wish resuscitation. Much anxiety and confusion regarding this topic existed and staff clearly required further education and guidance in order to develop policies within their units and to provide standard treatment within all specialist palliative care units. PMID- 15750520 TI - An evaluation of dependency assessment: experiences of staff, patients and carers in a UK hospice. AB - AIM: To evaluate the experiences of health-care professionals, service users and carers of service users from a UK hospice in relation to dependency assessment. DESIGN: Qualitative evaluation. SAMPLE: Purposive, convenience sample of hospice staff, patients receiving hospice services and carers of patients receiving hospice services. METHODS: Focus group interviews with hospice staff and carers of hospice service users. Individual interviews with patients. RESULTS: Staff felt the tool used produced inaccurate results, in part because of omissions in content. They did not perceive the data produced affected working practice, e.g. staffing levels. Patients described unobtrusive assessment and stated their needs were met. Carers reported feelings of isolation. They also described differing information and support needs from the patients. CONCLUSION: Further research is required to improve the validity of dependency data collection. Further investigation of carer assessment is also justified. PMID- 15750521 TI - How basic is palliative care? PMID- 15750522 TI - Palliative care needs assessment. PMID- 15750524 TI - Poverty and palliative care in the US: issues facing the urban poor. AB - Poverty is a significant public health and social problem in the US. The urban poor living with life-limiting illnesses are a particularly vulnerable population. The literature related to the experiences of the urban poor at the end of life is sparse. Most relates to the experiences of patients with cancer. The purpose of this literature review is to describe the problem of poverty in the US, to identify challenges in providing palliative care to the urban poor, and lastly, to articulate implications for nursing practice and nursing research. PMID- 15750525 TI - Learning disability and palliative care. PMID- 15750527 TI - An overview of the end-of-life discussion. AB - Good communication with patients can be a powerful tool in palliative care. However, discussing fears and anxieties at length is also something that many nurses find difficult. This article provides an overview of one approach to using the power of communication, the end-of-life discussion. Practical issues are considered, as well as suggestions as to what to include in such a discussion. Concepts of loss and meaning for the person with advanced disease are looked at. The article also considers some of the key communication skills that are especially valuable, such as active listening, ways of encouraging disclosure and the handling of silences. PMID- 15750526 TI - Learning disabilities and palliative care: recognizing pitfalls and exploring potential. AB - People with learning disabilities may experience poorer health, have additional health needs and often die younger than the average population. They may be diagnosed with palliative conditions but have difficulties accessing appropriate assessment, treatment and subsequent care. This position paper introduces the recognized challenges of providing high quality palliative care to people with learning disabilities and explores the potential for delivering appropriate palliative care for this vulnerable population within the UK. The article concludes with recommendations for practice development. PMID- 15750528 TI - A personal reflection on sitting at the beside of a dying loved one: the final hours of life. PMID- 15750529 TI - How basic is palliative care? Responses to Harmala Gupta. PMID- 15750531 TI - Clinical trials needed to answer questions about Invisalign. PMID- 15750532 TI - ABO resident clinical outcomes study: case complexity as measured by the discrepancy index. PMID- 15750533 TI - Multicenter, Internet-based orthodontic education: a research proposal. PMID- 15750535 TI - Physical properties of root cementum: Part 3. Hardness and elastic modulus after application of light and heavy forces. AB - BACKGROUND: The aims of this investigation were to evaluate the hardness and elastic modulus of untreated human premolar cementum and to investigate the changes after application of light and heavy orthodontic forces. METHODS: Thirty six maxillary and mandibular first premolars were collected from 16 prospective orthodontic patients. The patients were assigned to 1 of 2 groups. Group I (light force) consisted of 8 subjects (4 male, 4 female, mean age 14.8 years) who had 25 g of buccally directed orthodontic force applied to the first premolars on 1 side. Group II (heavy force) consisted of 9 subjects (7 male, 2 female, mean age 13.8 years) who had 225 g of buccally directed orthodontic force applied to first premolars on 1 side. Contralateral premolars served as the control in both groups. The force was applied for 4 weeks; then each first premolar was extracted and stored in deionized water. Physical properties were tested on unprepared cementum on buccal and lingual surfaces at the cervical, middle, and apical thirds with an ultra-micro-indentation system. RESULTS: Mean (+/- standard deviation) hardness of the untreated cementum at the cervical, middle, and apical thirds, respectively, was 0.25 +/- 0.09 GPa, 0.24 +/- 0.07 GPa, and 0.18 +/- 0.06 GPa on the buccal surface and 0.24 +/- 0.08 GPa, 0.24 +/- 0.06 GPa, and 0.21 +/- 0.06 GPa on the lingual surface. Mean elastic modulus at the cervical, middle, and apical thirds, respectively, was 4.4 +/- 2.4 GPa, 3.4 +/- 2.0 GPa, and 2.4 +/ 1.8GPa on the buccal surface and 3.8 +/- 2.1 GPa, 3.2 +/- 1.4 GPa, and 2.4 +/- 1.5 GPa on the lingual surface. CONCLUSIONS: The mean hardness and elastic modulus of untreated human premolar cementum gradually decreased from cervical to apical regions of the buccal and lingual surfaces. A similar decrease was observed in the experimental teeth of light-force and heavy-force groups. There were no statistically significant differences between the control and experimental teeth; this is probably due to large intraindividual variation. PMID- 15750536 TI - Physical properties of root cementum: Part 4. Quantitative analysis of the mineral composition of human premolar cementum. AB - BACKGROUND: The aim of this study was to perform a quantitative analysis of the calcium (Ca), phosphorus (P), and fluoride (F) concentrations in human first premolars. METHODS: This study was conducted on 18 maxillary or mandibular first premolars that were collected from 16 prospective orthodontic patients (10 male, 6 female), mean age 13.9 years (range, 11.7-16.1 years), requiring first premolar extractions. After extraction, the teeth were prepared for electron probe microanalysis. The Ca, P, and F concentrations were measured on the buccal and lingual surfaces at the midpoint of the cervical, middle, and apical thirds of the root from the outer to middle to inner third of the cementum. RESULTS AND CONCLUSIONS: In first premolar cementum, there was significant interindividual variation in the Ca, P, and F concentrations ( P = .024, .017, and .000, respectively). There was no significant difference in the Ca, P, and F concentrations of cementum between the buccal and lingual surfaces, except for a significantly higher F content at the cervical region on the buccal surface ( P = .000). There was a decreasing gradient in the Ca, P, and F concentrations from the cervical to the apical third of the root, which was highly significant from the cervical to middle third ( P = .000) and from the middle to apical third ( P = .000), except for F, for which there was no significant difference from the cervical to the middle third on the lingual surface ( P = .966). There was a significant increasing gradient in the Ca and P concentrations from the outer to inner third of cementum at the cervical ( P < .01) and middle ( P < .01) thirds of the root but no significant difference at the apical third of the root. For F, there was a significant decreasing gradient from the outer to the inner third of cementum at the cervical ( P < .01), middle ( P < .01) and apical ( P < .01) thirds of the root. PMID- 15750537 TI - Physical properties of root cementum: Part 5. Volumetric analysis of root resorption craters after application of light and heavy orthodontic forces. AB - BACKGROUND: In previous studies on root resorption, resorption was quantified by making histologic cuts or by surface area measurements of resorption craters. The aims of this study were to evaluate the effects of orthodontic force magnitude on root resorption craters with volumetric measurements and also to identify the sites that might be predisposed to resorption. METHODS: After an experimental time of 28 days, 36 human premolars previously divided into light-force (25 g) and heavy-force (225 g) groups were extracted and prepared for scanning electron microscope imaging. Pairs of stereo images were taken, and 3-dimensional quantitative volumetric analysis was performed with commercial software. RESULTS AND CONCLUSIONS: The mean volume of the resorption crater in the light-force group was 3.49-fold greater than in the control group; the mean volume of the resorption crater in the heavy-force group was 11.59-fold greater than in the control group ( P < .001). The heavy-force group had 3.31-fold greater total resorption volume than the light-force group ( P < .001). Buccal cervical and lingual apical regions had significantly more resorption craters than the other regions ( P < .001). There was more resorption by volume in the heavy-force group compared with the light-force group and controls. Although more resorption was recorded in the light-force group, the difference in the amount of resorption between the light-force and control groups was not statistically significant. There was significantly more resorption on the buccal cervical and lingual apical regions of the root surfaces than on the other regions, suggesting that high pressure zones might be more susceptible to resorption after 28 days of force application. PMID- 15750538 TI - Examining the role of the orthodontist in preventing adolescent tobacco use: a nationwide perspective. AB - PURPOSE: The purpose of this study was to determine the level of antitobacco practices currently in place in orthodontic offices across the United States, so that an antitobacco standard of care might be derived. METHODS: A 23-item survey was constructed and mailed to 200 orthodontists practicing in the United States, asking about antitobacco counseling and record keeping, concern for the matter, level of preparedness in helping a patient quit smoking, and potential barriers to effective antitobacco practices. RESULTS: A corrected response rate of 59.5% (n = 119) was obtained. Whereas 89.9% of respondents were concerned about tobacco use by their adolescent patients, only 50% reported actually asking their patients whether they use tobacco. Most orthodontists (67.5%) reported that they are either "not sure" or "not ready" to provide effective cessation counseling to patients who use tobacco, but 61.1% would be willing to integrate a tobacco control program into their practices. No orthodontists were familiar with the National Cancer Institute's strategy for doctors to help their patients stop tobacco habits, called the "Five A's" (formerly the "Four A's"). CONCLUSION: Because of the unique and often positive interactions orthodontists have with their adolescent patients, members of the specialty can play significant roles in educating patients about the health effects of tobacco use. Because of the lack of adequate training, this education is not taking place in orthodontic practices in the United States. PMID- 15750539 TI - Heritability of craniofacial characteristics between parents and offspring estimated from lateral cephalograms. AB - OBJECTIVE: The purpose of the study was to estimate the heritability of different cephalometric parameters, according to lateral cephalograms, between parents and their offspring in an Icelandic population. METHODS: The material was collected at the Faculty of Odontology, University of Iceland. The subjects were 363 children (6 years of age) and their parents. Material was also collected from the same group of children at the age of 16 years. Twenty-two reference points were identified on each cephalogram, and 33 variables were calculated, both angular and linear. Heritability was calculated at ages 6 and 16. RESULTS: Daughters had more variables that reached the level of significance than did sons. Daughters showed similar heritability to both parents at both age levels, but more variables were highly significant ( P < or = .001) in the daughter-father groups. Sons showed stronger heritability to their mothers at both ages. The variables showing the greatest heritability were those representing the position of the lower jaw, the anterior and posterior face heights, and the cranial base dimensions. Heritability was notably low for the dental variables. CONCLUSIONS: Genetic aberrations can be detected for complex polygenetic multifactorial traits. Cephalometric data can support predictions, and analysis of parental data could have predictive value for offspring. PMID- 15750540 TI - Buccal corridors and smile esthetics. AB - PURPOSE: The purpose of this study was to determine the influence of buccal corridors on smile attractiveness when judged by lay persons. MATERIAL: Full-face color slides of 10 randomly selected smiling subjects (5 women, 5 men) were digitized. The maxillary posterior dentitions for all subjects were digitally altered to produce a range of smile fullness: narrow (28% buccal corridor), medium-narrow (22% buccal corridor), medium (15% buccal corridor), medium-broad (10% buccal corridor), and broad (2% buccal corridor). The 5 images of each subject were paired into 11 possible combinations, and the resulting 110 pairings were randomly projected to a panel of 30 adult lay persons who compared the 2 images in each pair for smile attractiveness. RESULTS: Statistical analysis with the Wilcoxon signed-rank and rank-sum tests showed that (1) a broader smile (minimal buccal corridor) was judged by lay persons to be more attractive than a narrow smile (larger buccal corridors), and (2) no significant differences were found in judging between male and female subjects or between male and female judges. CONCLUSIONS: Having minimal buccal corridors is a preferred esthetic feature in both men and women, and large buccal corridors should be included in the problem list during orthodontic diagnosis and treatment planning. PMID- 15750541 TI - Botulinum toxin type A in the treatment of excessive gingival display. AB - PURPOSE: One cause of excessive gingival display is the muscular capacity to raise the upper lip higher than average. Several surgical procedures have been reported to improve the condition, but surgery always involves risk and is costly. Botulinum toxin type A (BTX-A) (Botox; Allergan, Irvine, Calif) has been studied since the late 1970s for the treatment of several conditions associated with excessive muscle contraction or pain. This clinical pilot study was performed to determine whether BTX-A injections would reduce excessive gingival display. MATERIAL: Five subjects with excessive gingival display due to hyperfunctional upper lip elevator muscles were treated with BTX-A injections. RESULTS: This treatment modality was effective, producing esthetically acceptable smiles in these patients. The improvements lasted 3 to 6 months. CONCLUSIONS: Injection with BTX-A at preselected sites is a novel, cosmetically effective, minimally invasive alternative for the temporary improvement of gummy smiles caused by hyperfunctional upper lip elevator muscles. PMID- 15750542 TI - Evaluation of posttreatment changes in Class II Division 1 patients after nonextraction orthodontic treatment: cephalometric and model analysis. AB - BACKGROUND: The aim of this retrospective study was to evaluate changes in patients with Class II Division 1 malocclusions treated with cervical headgear and full fixed orthodontic appliances and without extractions. METHODS: Lateral cephalograms and dental casts of 18 patients were obtained at the beginning and the end of treatment and at the postretention period. Pretreatment records were taken at 11.9 +/- 2.6 years, and the mean treatment time was 2.8 +/- 1.2 years. All patients were asked to use maxillary and mandibular Hawley retainers for 1 year. The average postretention interval was 5.3 years. RESULTS: The assessment of cephalometric findings showed that maxillary growth was restricted; maxillary incisor inclination, overjet, and overbite were decreased; the mandibular incisors were proclined; and a slight mandibular posterior rotation occurred at the end of treatment. At the postretention period, the maxilla remained stable, the mandible moved anteriorly, and the overjet increased slightly (1.25 mm, P < .01). The dental cast changes indicated that both maxillary and mandibular irregularity index values decreased significantly during treatment but increased after 5 years. Mandibular intercanine width decreased 0.47 mm ( P < .01) during the postretention period. CONCLUSIONS: The redirection of maxillary growth remained stable in patients with Class II malocclusions treated with cervical headgear and fixed orthodontic appliances and without extractions, but treatment induced mandibular rotations tended to return to their original patterns. Various dental changes occurred during the postretention period. PMID- 15750543 TI - Combining orthodontic tooth movement with distraction osteogenesis to close cleft spaces and improve maxillary arch form in cleft lip and palate patients. PMID- 15750544 TI - Orthodontic treatment of a traumatically intruded tooth with ankylosis by traction after surgical luxation. AB - This report presents the treatment of a patient with anterior crossbite and displacement of ankylosed maxillary and mandibular right lateral incisors. The maxillary lateral incisor, which was traumatically intruded, was successfully treated with 2 surgical luxations followed by orthodontic traction. The mandibular lateral incisor, which was avulsed and replanted, experienced replacement root resorption during orthodontic treatment and was ultimately extracted. Two years after active orthodontic treatment, the occlusal results were considered satisfactory. PMID- 15750545 TI - Category 5: Class II Division 1 malocclusion. AB - This case report was part of a display of cases sponsored by the ABO after the board-certification process. The Summary of Treatment and records are reprinted here with minimal editing and reformatting, so the presentation resembles the actual documents submitted to the board. PMID- 15750546 TI - Evidence-based versus experience-based views on occlusion and TMD. PMID- 15750547 TI - Orthodontics in 3 millennia. Chapter 1: Antiquity to the mid-19th century. AB - Orthodontics had its beginnings in the time of the ancient Egyptians, who used crude metal bands and catgut, but it was not until the late 18th century that the first practical appliances came into use. These were fine-tuned during the early 1900s; today's mechanisms are merely refinements. Major changes occurred when practitioners--originally physicians--began turning their attention from cosmetic "regulating" to occlusion and stability, while empiricism gave way to objectivity and the scientific method. The purpose of this article is to review the history of orthodontics from antiquity to the modern era. The article is divided into chapters that will be presented serially in every other issue of the Journal. PMID- 15750548 TI - Principles of cosmetic dentistry in orthodontics: Part 3. Laser treatments for tooth eruption and soft tissue problems. PMID- 15750549 TI - Patient abandonment. PMID- 15750551 TI - Barriers to effective pediatric asthma care. AB - Although progress has been made in understanding the pathophysiology of asthma and identifying key features of quality asthma care, the prevalence of childhood asthma remains high. Barriers to effective asthma care that currently exist include the persistence of environmental risk factors, disparities in care that stem from poverty and cultural differences, and inconsistencies in the quality of asthma care provided by clinicians. Pediatric nurse practitioners at Yale New Haven Children's Hospital have actively implemented the recommended guidelines for asthma care and addressed causes for some of the disparities in asthma health care. Two major initiatives are described: the Asthma Care Coordination Project at Yale New Haven Hospital Pediatric Primary Care Center, and the establishment of an Asthma Outreach Program. Recommended resources and Web sites for the practitioner are also provided. PMID- 15750552 TI - Research endeavors among pediatric nurse practitioners (REAP) study. AB - INTRODUCTION: The purpose of this study was to identify the facilitators of and barriers to research utilization and to identify research activities and resources needed to promote research among pediatric nurse practitioners. METHOD: A random sample of 10% of the NAPNAP membership (n = 627) was surveyed with a response rate of 69% (n = 431). The survey included the BARRIERS Scale to identify facilitators of and barriers to research utilization, questions about research activities, and resources needed to promote research. RESULTS: Barriers to research included time constraints for reading and implementing research, the amount of research information and the way it is compiled, and knowledge deficits about statistical analysis interpretation. Facilitators of research utilization included having time to review and implement findings, authority to change nursing practice based on research findings, and conducting clinically relevant studies. Time constraints (82%), lack of grant writing skills (66%), and funding (57%) were identified as barriers to conducting research. DISCUSSION: Findings from this study will be used by the National NAPNAP Research Committee to plan and implement programs to increase the utilization of nursing research in clinical practice and provide guidance, education, and support to promote involvement in research activities among the membership. PMID- 15750553 TI - Health and social context: social capital's utility as a construct for nursing and health promotion. AB - Social capital is a health-related construct that is a way for pediatric nurse practitioners (PNPs) to conceptualize the interconnectedness between health and social relations. Applying the concept of "social capital" to child and family health has the capacity to help PNPs reframe persistent questions and explore health issues in new and creative ways. A health-enabling environment must be built using a bottom-up approach, with participatory health promotion programs and internally driven community health projects. PNPs must create opportunities for networking and diffusing knowledge about community-related health issues. PNPs also must build their own social capital and practice "vision therapy" to develop new interventions to enhance child and family health. PMID- 15750554 TI - Relationships between physical symptoms and pubertal development. AB - INTRODUCTION: The purpose of this study was to examine relationships between physical symptoms and pubertal status and timing in adolescents. METHOD: This descriptive study was based on cross-sectional analysis of survey data from the National Longitudinal Study of Adolescent Health including a school-based sample of more than 20,000 adolescents in grades 7 through 12. Self-reported physical symptoms during the past 12 months were examined. Pubertal development was measured as pubertal status (early, mid, and advanced pubertal) and timing (early timing, on time, and late timing). RESULTS: Headaches and musculoskeletal pains were the most commonly reported symptoms among advanced pubertal status respondents. Headache was more prominent among girls; among boys, musculoskeletal complaints were predominant. For both sexes, on-time developers were less likely to report physical symptoms when compared with the two off-timing groups. Among girls, pubertal status and timing were more closely associated with various types of symptoms. DISCUSSION: This study underscores the importance of both the objective signs of pubertal changes and youngsters' own perception of their pubertal changes in assessing and addressing physical symptoms. PMID- 15750555 TI - The use of focus groups in pediatric and adolescent research. AB - Through this past decade, health care research has focused on clinical trials and establishing intervention versus control group research to improve the health of children and adolescents. Many of these clinical intervention trials have not demonstrated significantly improved health for these persons. One possible reason for the failure to demonstrate improved health is that the young persons' perspective is lacking. Focus group method of data collection is an excellent vehicle to help gain the young person's perspective on a variety of issues related to health and wellness. Including the young person's perspective when designing a program may encourage this person to participate in the new initiative, which would ultimately improve the health of children and adolescents to a greater degree than when their perspective is not taken into account. This article discusses the use of focus groups as an important part of the research process in pediatric and adolescent populations. PMID- 15750556 TI - Hydrocephalus. PMID- 15750557 TI - The current medical malpractice environment: an analysis of causes and solutions. PMID- 15750558 TI - The acute care pediatric nurse practitioner. PMID- 15750559 TI - Asthma medication delivery devices. PMID- 15750566 TI - Reducing neonatal mortality rate with nurse-midwives. AB - Newborn health indices were measured before, during, and after a demonstration nurse-midwife program introduced to relieve a health manpower shortage at a county hospital in rural California. During the program, prenatal care increased, and prematurity and neonatal mortality rate decreased at the county hospital. After the 3 year program, prenatal care decreased while prematurity rose from 6.6 to 9.8 per cent (p<0.02) and neonatal mortality rate rose from 10.3 to 32.1 per 1,000 live births (p<0.005). No significant changes occurred in the same indices for births elsewhere in the county throughout the period of the study. It is concluded that the discontinuation of the nurse-midwives' services was the major factor in these changes, and it is suggested that nurse-midwives be used more extensively. PMID- 15750576 TI - Bioassay of various pesticides by microcalorimetry measuring the metabolic heat of yeast. AB - Using a microcalorimeter, the heat evolved during incubation of yeast cultures at 30 degrees C was detected in the form of a growth thermogram (metabolic heat incubation time curve). The correlation between the growth thermograms and the growth curves obtained by colony counting or from the turbidity measurements of the cultures was very good. In this study, the effects of 12 pesticides that are used widely on golf courses on the growth thermograms of yeast were investigated and the inhibitory action of the pesticides on the growth of yeast was quantitatively evaluated using a microcalorimeter (Biothermo Analyzer). The growth thermograms were applied to estimate the microbial activity of yeast in the presence of pesticide and to determine the 50% inhibitory concentration (K) and minimum inhibitory concentration (MIC), which can be regarded as good indexes of the toxic potency of pesticides. It was found that the addition of TPN, captan, thiuram and copper oxide to the yeast cultures clearly induced inhibitory action on the growth of yeast cells. On the other hand, the other 8 pesticides, iprodione, isoprothiorane, chloroneb, pencycuron, metalaxyl, toleclofos-methyl, flutolanil, and mepronil did not affect the growth over the concentration range up to 30 mg/l. PMID- 15750577 TI - Genomic profile of roundup treatment of yeast using DNA microarray analysis. AB - The herbicide Roundup, which contains glyphosate as the active ingredient, was first introduced in 1974 and has enjoyed widespread use in Japan and elsewhere in the world. Roundup-induced reactions occurring in the yeast Saccharomyces cerevisiae may have a predictive value for understanding responses in higher eukaryotes, and we applied yeast DNA microarray analysis for this purpose. Functional characterization of up-regulated open reading frames (ORFs) following Roundup treatment suggests that Roundup affects membrane structures and cellular organelles. Expression profiles induced by treatments with detergents, oils and hydrostatic pressure were similar to those following Roundup treatment based on cluster analysis. Glyphosate alone was not found to inhibit yeast growth at the concentration contained in the Roundup treatment used for microarray analysis. The toxicity of Roundup appeared to be due to detergent in the product. PMID- 15750578 TI - Construction of a data dependent analysis (DDA) yeast cDNA microarray experiment for use in toxicogenomics. AB - DNA microarrays are becoming increasingly popular in toxicogenomic studies. It is common knowledge that multiple repeats of microarray experiments have to be performed with multiple samples to get reliable data, due to experimental variations (both technical and biological) contained in microarray experiments. However, the use of multiple samples and replicate experiments is not practical in the field of environmental toxicology, since environmental samples are limited and microarray experiments are expensive. Thus it is desirable to obtain reliable data from a minimum number of repeats of microarray experiments. To further establish and extend gene expression profiling for toxicogenomics using microarrays, it is necessary to establish an analytical method of microarrays for toxicogenomics. In this study, we attempted to construct a "Data Dependent Analysis (DDA)" yeast cDNA microarray experiment from 100 microarray datasets in order to get reliable data from one microarray experiment. PMID- 15750579 TI - Fluoride concentration in superficial enamel layers of permanent teeth in children residing in areas with different levels of environmental pollution with fluorine compounds. AB - The concentration of fluoride in superficial layers of enamel of permanent teeth was studied in 14-year-old children residing in Szczecin and Bialystok-two cities differing regarding the level of environmental fluorine pollution. It was found that higher environmental pollution with fluorine favors the accumulation of this element in superficial layers of enamel in the form of structural fluoride. Total and structural fluoride concentration decreased with layer depth. PMID- 15750580 TI - Isolation of cDNA of novel cytochrome P450 1B gene, CYP1B2, from Carp (Cyprinus carpio) and its induced expression in gills. AB - Cytochrome P450 (CYP) enzymes constitute a multigene family of many endogenous and xenobiotic substances. The CYP1 family is of particular interest in environmental toxicology because its members are dominant in the metabolism of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and aryl amines. A new complementary DNA of the CYP1B subfamily encoding CYP1B2 was isolated from carp liver after intraperitoneal injection with 3 methylcholanthrene (3-MC). The obtained full-length cDNA contained a 5'noncoding region of 161 bp, an open reading frame of 1593 bp coding for 530 amino acids and a stop codon, and a 3'noncoding region of 1457 bp. The predicted molecular weight of the protein was approximately 60.2 kDa. The deduced amino acid sequence of this cDNA was 91% similar to that of our previously reported carp CYP1B1; its similarities with those of the reported CYP1B1s of teleosts and mammals were 60.8, 54.4, 50.8, and 51.4% for plaice, human, rat, and mouse, respectively. The phylogenetic tree of fish and mammalian CYP1 sequences constructed by the protein maximum-likelihood method suggested a relatively recent divergence of CYP1B2 from CYP1B1 in the ancestor of carp and closely related species. Despite the structural similarity of CYP1B2 with CYP1B1, which showed induced expression in 3 MC-treated liver, intestine, and gills with marked constitutive expression in gills, CYP1B2 revealed induced expression in gills but not in liver or intestine, and no detectable constitutive expression in the tissues studied. PMID- 15750581 TI - Implementation of bioavailability in standard setting and risk assessment. AB - Bioavailability is, to an increasing extent, recognised as the key issue linking the increased levels of toxicants to actually occurring adverse effects in ecosystems, whilst taking the modifying effects of the abiotic components of the environment into account. Various factors may affect bioavailability in the field, and these factors are often time and space dependent. This is one of the main reasons why legislators have been reluctant to implement bioavailability in risk assessment procedures. Over the last few years, however, considerable scientific progress has been made with regard to increasing our understanding of the chemical and ecological mechanisms responsible for rendering chemicals available for uptake and toxicity. As a consequence, legislators face the challenge of having to anticipate the scientific progress and to implement bioavailability in legislation. This contribution reports on the possibilities of implementing various methodologies within a maximum time period of three years. PMID- 15750582 TI - Social factors associated with psychological distress among inhabitants in a methyl-mercury polluted area in rural Japan. AB - Little attention has been paid to the mental health of inhabitants of methylmercury-polluted areas in Japan. This study examined the relationship between one's experience with Minamata disease (MD) (such as compensation issues) and psychological distress. The subjects were 133 (44.2%) of the 301 inhabitants over the age of 40 years living in two fishing village districts along the coast of the Yatsushiro Sea which had been contaminated with methylmercury. Data on the inhabitants' experience with MD, social network factor, health condition and mental health were obtained using questionnaires including the General Health Questionnaire (GHQ)-30. The proportional odds model was used to estimate the adjusted odds ratios of factors associated with a higher GHQ score after adjustment for age, sex and village. MD status based on MD compensation, level of participation in MD patients' groups, and presence of certified MD patients in the family were significantly associated with psychological distress. Although these associations decreased after further adjustments were made taking health condition into consideration, MD status, participation in several sit-ins and the presence of certified MD patients in the family maintained marginally positive association with psychological distress. Further investigations with more precise and detailed measurements are needed to corroborate the relationship between inhabitants' experience with MD and mental health. PMID- 15750583 TI - Transient inhibition of synergistically insulin-like growth factor-1- and bisphenol A-induced poliferation of estrogen receptor alpha (ERalpha)-positive human breast cancer MCF-7 cells by melatonin. AB - There are many subclones of human breast cancer MCF-7 cells that respond to different levels of estrogen and that have been used for evaluating the estrogenic potential of environmental chemicals such as bisphenol A. In this study, I examined the effects of melatonin, an endogenous growth-inhibitory hormone, on the bisphenol A-induced proliferation of an MCF-7 subclone, designated as MCF-7-EMF cells. MCF-7-EMF cell growth was extremely slow in the presence of either estrogen at 10(-7) M or bisphenol A at 10(-7) M in a phenol red-free medium and 10% charcoal-stripped fetal bovine serum. It was difficult to detect the increment of BrdU incorporation in MCF-7-EMF cells (2x10(4) cells) upon exposure to 10(-7) M bisphenol A alone for up to 50 h. The same result was obtained with cells exposed to 10 ng/ml insulin-like growth factor-1 (IGF-1) alone for up to 50 h. However, in the presence of 10 ng/ml IGF-1, bisphenol A markedly increased cell proliferation. The synergistic response was transiently inhibited by 10(-10) M melatonin (p<0.05). Thus, melatonin caused a transient inhibition of cell growth of estrogen receptor alpha (ERalpha)-positive MCF-7-EMF cells induced synergistically by both IGF-1 and bisphenol A. PMID- 15750584 TI - Separation and characterization of alkyltrimethylbenzene derivatives in diesel exhaust particles (DEP). AB - In our continuing studies on estrogenic compounds in diesel exhaust particles (DEP), we have reported the systematic separation of alkyldibenzothiophenes from the estrogenic hexane fraction of DEP. In this study, another estrogenic fraction was further isolated and characterized. DEP were roughly fractionated by successive extraction with hexane, benzene, dichloromethane, methanol, 1 M ammonia and 1 M HCl. The hexane extract, which exhibited estrogenic activity, was further fractionated by column chromatography and high performance liquid chromatography (HPLC). From one of the obtained fractions that showed estrogenic activity, three alkyltrimethylbenzenes were identified by gas chromatography-mass spectrometry (GC-MS) and 1H-nuclear magnetic resonance (NMR) spectroscopy. PMID- 15750585 TI - Arsenicosis in two villages in Terai, lowland Nepal. AB - Although arsenic contamination of groundwater has been detected recently in the Terai region of Nepal, scarce data have been available regarding the possible health effects of the contamination. A study was carried out in two rural villages of Nawalparasi district, where the existence of arsenic contamination has been reported. Almost all tubewells in one of the two villages (Goini, G) exceeded (ranging from 0.104 mg/L to 1.702 mg/L) the maximum permissible limit for arsenic in drinking water in Nepal (0.05 mg/L), and only a few tubewells (19.5%) in the other village (Kunuwar Big, KB) were below this level (0.004 mg/L to 0.972 mg/L). Prevalence rates of arsenicosis diagnosed on the basis of the presence of dermatological manifestations were 11.3% (56 out of 495 examined) and 6.5% (34/525) in G and KB, respectively, with an overall prevalence rate of 8.9%. In terms of gender, the prevalence rate was higher in males (11.5%; 61/531) than in females (5.9%; 29/489), supporting previous observations including our own. Most (71.1%) of the identified patients were 20-49 years old, and the prevalence rate appeared to be higher among those in their 30s or 40s in the both sexes. The youngest patient was 10 years of age. All the patients were either in the mild or moderate stage of the disease and most (78.9%) were in the mild stage. Melanosis with some other manifestations was observed in 95.6% of the patients, while keratosis was found in 57.8%. Leucomelanosis without any other manifestations, which has not been reported either in Bangladesh or India, was observed in 3.3% of the identified patients. Only one patient had Bowen's disease. To our knowledge, the current study provides for the first time quantitative information regarding the arsenicosis patients in Nepal. PMID- 15750586 TI - A dominant lethal genetic system for autocidal control of the Mediterranean fruitfly. AB - The Sterile Insect Technique (SIT) used to control insect pests relies on the release of large numbers of radiation-sterilized insects. Irradiation can have a negative impact on the subsequent performance of the released insects and therefore on the cost and effectiveness of a control program. This and other problems associated with current SIT programs could be overcome by the use of recombinant DNA methods and molecular genetics. Here we describe the construction of strains of the Mediterranean fruit fly (medfly) harboring a tetracycline repressible transactivator (tTA) that causes lethality in early developmental stages of the heterozygous progeny but has little effect on the survival of the parental transgenic tTA insects. We show that these properties should prove advantageous for the implementation of insect pest control programs. PMID- 15750587 TI - Role of the RB1 family in stabilizing histone methylation at constitutive heterochromatin. AB - Here, we show a role for the RB1 family proteins in directing full heterochromatin formation. Mouse embryonic fibroblasts that are triply deficient for RB1 (retinoblastoma 1), RBL1 (retinoblastoma-like 1) and RBL2 (retinoblastoma like 2) - known as TKO cells - show a marked genomic instability, which is coincidental with decreased DNA methylation, increased acetylation of histone H3 and decreased tri-methylation of histone H4 at lysine 20 (H4K20). Chromatin immunoprecipitation showed that H4K20 tri-methylation was specifically decreased at pericentric and telomeric chromatin. These defects are independent of E2F family function. Indeed, we show a direct interaction between the RB1 proteins and the H4K20 tri-methylating enzymes Suv4-20h1 and Suv4-20h2, indicating that the RB1 family has a role in controlling H4K20 tri-methylation by these histone methyltransferases. These observations indicate that the RB1 family is involved in maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin, linking tumour suppression and the epigenetic definition of chromatin. PMID- 15750588 TI - Gaze fixation and the neural circuitry of face processing in autism. AB - Diminished gaze fixation is one of the core features of autism and has been proposed to be associated with abnormalities in the neural circuitry of affect. We tested this hypothesis in two separate studies using eye tracking while measuring functional brain activity during facial discrimination tasks in individuals with autism and in typically developing individuals. Activation in the fusiform gyrus and amygdala was strongly and positively correlated with the time spent fixating the eyes in the autistic group in both studies, suggesting that diminished gaze fixation may account for the fusiform hypoactivation to faces commonly reported in autism. In addition, variation in eye fixation within autistic individuals was strongly and positively associated with amygdala activation across both studies, suggesting a heightened emotional response associated with gaze fixation in autism. PMID- 15750590 TI - Control of synaptic strength and timing by the release-site Ca2+ signal. AB - Transmitter release is triggered by highly localized, transient increases in the presynaptic Ca(2+) concentration ([Ca(2+)]). Rapidly decaying [Ca(2+)] elevations were generated using Ca(2+) uncaging techniques, and [Ca(2+)] was measured with a low-affinity Ca(2+) indicator in a giant presynaptic terminal, the calyx of Held, in rat brain slices. The rise time and amplitude of evoked excitatory postsynaptic currents (EPSCs) depended on the half-width of the fluorescence transient, which was predicted by a five-binding site model of a Ca(2+) sensor having relatively high affinity (K(d) approximately 13 microM). Very fast [Ca(2+)] transients (half-width <0.5 ms) evoked EPSCs similar to those elicited by a single action potential (AP) in the same synapse. Triggering release with dual [Ca(2+)] transients of variable amplitudes demonstrated the supralinear transfer function of the sensor. The sensitivity of release to the time course of the [Ca(2+)] transient may contribute to mechanisms by which the presynaptic AP waveform controls synaptic strength. PMID- 15750589 TI - Vasoactive intestinal polypeptide mediates circadian rhythmicity and synchrony in mammalian clock neurons. AB - The mammalian suprachiasmatic nucleus (SCN) is a master circadian pacemaker. It is not known which SCN neurons are autonomous pacemakers or how they synchronize their daily firing rhythms to coordinate circadian behavior. Vasoactive intestinal polypeptide (VIP) and the VIP receptor VPAC(2) (encoded by the gene Vipr2) may mediate rhythms in individual SCN neurons, synchrony between neurons, or both. We found that Vip(-/-) and Vipr2(-/-) mice showed two daily bouts of activity in a skeleton photoperiod and multiple circadian periods in constant darkness. Loss of VIP or VPAC(2) also abolished circadian firing rhythms in approximately half of all SCN neurons and disrupted synchrony between rhythmic neurons. Critically, daily application of a VPAC(2) agonist restored rhythmicity and synchrony to VIP(-/-) SCN neurons, but not to Vipr2(-/-) neurons. We conclude that VIP coordinates daily rhythms in the SCN and behavior by synchronizing a small population of pacemaking neurons and maintaining rhythmicity in a larger subset of neurons. PMID- 15750592 TI - Modularity and evolutionary constraint on proteins. AB - Modularity, which has been found in the functional and physical protein interaction networks of many organisms, has been postulated to affect both the mode and tempo of evolution. Here I show that in the yeast Saccharomyces cerevisiae, protein interaction hubs situated in single modules are highly constrained, whereas those connecting different modules are more plastic. This pattern of change could reflect a tendency for evolutionary innovations to occur by altering the proteins and interactions between rather than within modules, in a manner somewhat similar to the evolution of new proteins through the shuffling of conserved protein domains. PMID- 15750591 TI - LAR receptor protein tyrosine phosphatases in the development and maintenance of excitatory synapses. AB - Leukocyte common antigen-related (LAR) family receptor protein tyrosine phosphatases (LAR-RPTP) bind to liprin-alpha (SYD2) and are implicated in axon guidance. We report that LAR-RPTP is concentrated in mature synapses in cultured rat hippocampal neurons, and is important for the development and maintenance of excitatory synapses in hippocampal neurons. RNA interference (RNAi) knockdown of LAR or dominant-negative disruption of LAR function results in loss of excitatory synapses and dendritic spines, reduction of surface AMPA receptors, impairment of dendritic targeting of the cadherin-beta-catenin complex, and reduction in the amplitude and frequency of miniature excitatory postsynaptic currents (mEPSCs). Cadherin, beta-catenin and GluR2/3 are tyrosine phosphoproteins that coimmunoprecipitate with liprin-alpha and GRIP from rat brain extracts. We propose that the cadherin-beta-catenin complex is cotransported with AMPA receptors to synapses and dendritic spines by a mechanism that involves binding of liprin-alpha to LAR-RPTP and tyrosine dephosphorylation by LAR-RPTP. PMID- 15750593 TI - Frequent somatic mutations of the transcription factor ATBF1 in human prostate cancer. AB - Cancer often results from the accumulation of multiple genetic alterations. Although most malignancies are sporadic, only a small number of genes have been shown to undergo frequent mutations in sporadic cancers. The long arm of chromosome 16 is frequently deleted in human cancers, but the target gene for this deletion has not been identified. Here we report that ATBF1, which encodes a transcription factor that negatively regulates AFP and MYB but transactivates CDKN1A, is a good candidate for the 16q22 tumor-suppressor gene. We narrowed the region of deletion at 16q22 to 861 kb containing ATBF1. ATBF1 mRNA was abundant in normal prostates but more scarce in approximately half of prostate cancers tested. In 24 of 66 (36%) cancers examined, we identified 22 unique somatic mutations, many of which impair ATBF1 function. Furthermore, ATBF1 inhibited cell proliferation. Hence, loss of ATBF1 is one mechanism that defines the absence of growth control in prostate cancer. PMID- 15750594 TI - Positional identification of TNFSF4, encoding OX40 ligand, as a gene that influences atherosclerosis susceptibility. AB - Ath1 is a quantitative trait locus on mouse chromosome 1 that renders C57BL/6 mice susceptible and C3H/He mice resistant to diet-induced atherosclerosis. The quantitative trait locus region encompasses 11 known genes, including Tnfsf4 (also called Ox40l or Cd134l), which encodes OX40 ligand. Here we report that mice with targeted mutations of Tnfsf4 had significantly (P F1 graft-versus-host reactions blocked the effect of early antigen exposure. Vaccination of transplant recipients against microbial or leukemia antigens may be worthy of study. PMID- 15750608 TI - A multidisciplinary treatment strategy that includes high-dose chemotherapy for metastatic retinoblastoma without CNS involvement. AB - The prognosis of patients with metastatic retinoblastoma is poor with conventional chemotherapy and radiation. Since retinoblastoma is highly chemosensitive, dose-escalation of chemotherapeutic agents with stem cell support should be promising. We report our experience with high-dose chemotherapy (HDC) and autologous stem cell transplantation (SCT) in patients with metastatic retinoblastoma. Five patients with metastatic retinoblastoma underwent HDC with autologous SCT following conventional chemotherapy and local radiation therapy. Stem cells (bone marrow in four and peripheral blood stem cells in one) were collected after marrow involvement was cleared. Melphalan was a key drug in all patients, and was administered in combination with other agents such as cisplatin, cyclophosphamide, carboplatin or thiotepa. Three patients are currently alive disease-free at 113, 107 and 38 months, respectively, from the time of SCT. They had no central nervous system (CNS) involvement. The two patients who died of disease had CNS involvement. No long-term sequelae of HDC have been noted. Our treatment strategy using HDC appears to be effective for treating metastatic retinoblastoma without CNS involvement. PMID- 15750609 TI - Evidence for a qualitative hierarchy within the Hoechst-33342 'side population' (SP) of murine bone marrow cells. AB - In vitro cobblestone area (CA)-forming cell (CAFC) and in vivo (short-term and competitive repopulation) assays demonstrate that a qualitative hierarchy exists within the Hoechst-33342-defined side population (SP) in murine bone marrow (BM). Consistent with and extending previous studies, we demonstrate that (i) hematopoietic activity found in whole BM (WBM) is concentrated within the SP, rather than the non-SP (NSP); and (ii) within the SP, those cells that more strongly efflux the dye (lower SP, LSP) are qualitatively different from those that less strongly efflux the dye (upper SP, USP). Qualitative differences are highlighted by evidence that (i) CA derived from LSP CAFC persist in culture significantly longer than CA derived from USP CAFC; (ii) short-term, multilineage repopulation of lethally irradiated mice by LSP cells is more rapid than that in mice receiving USP, NSP, whole SP (WSP), or WBM cells and (iii) LSP cells out compete USP cells in the multilineage hematopoietic repopulation of lethally irradiated recipients. These data suggest that LSP cells are of higher quality than USP cells and potentially provide a means by which qualitative changes in primitive hematopoietic progenitors occurring naturally with aging, or clinically as a consequence of therapeutic manipulation, can be assessed. PMID- 15750610 TI - Early measles vaccination in bone marrow transplant recipients. AB - Measles vaccination has been recommended after the second year following bone marrow transplant (BMT) in patients not receiving immunosuppressive drugs. During a measles outbreak, we vaccinated all patients after the first year of transplant, and conducted a prospective trial to evaluate safety, effectiveness and sustained immunity after early vaccination. Patients received attenuated virus vaccine between 9 and 18 months after BMT. A total of 51 patients were evaluated and 27 of them (52.9%) were receiving immunosuppressive drugs. Only mild adverse reactions were noted. Nine patients (17.6%) were susceptible (IgG< or =100 mIU/ml) at vaccination, and all seroconverted. In those immune at vaccination, a four-fold increase in measles IgG titers was found in one of 34 patients (2.9%) with specific IgG> or =200 mIU/ml compared to 14 of 17 (82.3%) with IgG<200 mIU/ml (P< 0.0001). Sustained immunity after 24 months was more likely to occur in patients with specific IgG levels< or =200 or > or =500 mIU/mL (83.4 and 100%, respectively) in comparison to patients with 20065%) of missense mutation at codon 235 in the DNA-binding Ets domain of PU.1 in murine AML. Earlier studies, outside the context of malignancy, determined that conversion of arginine 235 (R235) to any other amino-acid residue leads to ablation of DNA-binding function and loss of expression of downstream targets. We show that mutation of R235 does not lead to protein loss, and occurs specifically in those AMLs showing loss of one copy of PU.1 (P=0.001, Fisher's exact test). PU.1 mutations were not found in the coding region, UTRs or promoter of human therapy-related AMLs. Potentially regulatory elements upstream of PU.1 were located but no mutations found. In conclusion, we have identified the cause of murine radiation-induced AML and have shown that loss of one copy of PU.1, as a consequence of flanking radiation-sensitive fragile domains on chromosome 2, and subsequent R235 conversion are highly specific to this mouse model. Such a mechanism does not operate, or is extremely rare, in human AML. PMID- 15750629 TI - Promotion of mammary cancer development by tamoxifen in a mouse model of Brca1 mutation-related breast cancer. AB - Loss of full-length Brca1 in mammary epithelial cells of the mouse mammary tumor virus (MMTV)-Cre Brca1 conditional exon 11 deletion mouse model results in the development of mammary adenocarcinomas with similar genetic changes to those found in human BRCA1-mutation-related breast cancers. We used this experimental model to evaluate the chemopreventive effect of tamoxifen on the development of mammary preneoplasia and adenocarcinoma. No protective effects of tamoxifen administration on mammary cancer development were found. Instead, tamoxifen treatment significantly increased rates of mammary epithelial cell proliferation and the prevalence of mammary hyperplasia at 6 months of age. Tamoxifen-exposed mice developed adenocarcinomas at younger ages than control mice and a higher percentage of mice developed adenocarcinomas by 12 months of age. Both whole mouse and tissue culture cell models were used to test if loss of full-length Brca1 was associated with a relative increase in the agonist activity of tamoxifen. Tamoxifen induced increased ductal growth in MMTV-Cre Brca1 conditional mice compared to wild type. Estrogen receptor alpha (ERalpha) expression was downregulated in the tamoxifen-induced hyperplasias. Reducing BRCA1 levels in MCF-7 cells using siRNA resulted in a relative increase in the agonist activity of tamoxifen. Results suggest a model of mammary cancer progression in which loss of full-length Brca1 altered the agonist/antagonist activity of tamoxifen, resulting in tamoxifen-induced mammary epithelial cell proliferation with subsequent loss of ERalpha expression and development of ERalpha-negative hyperplasias and adenocarcinomas. PMID- 15750631 TI - Cell-autonomous induction of functional tumor suppressor 15-lipoxygenase 2 (15 LOX2) contributes to replicative senescence of human prostate progenitor cells. AB - Normal human prostatic (NHP) epithelial cells undergo senescence in vitro and in vivo, but little is known about the tissue-specific molecular mechanisms. Here we first characterize young primary NHP cells as CK5(+)/CK18(+) intermediate basal cells that also express several other putative stem/progenitor cell markers including p63, CD44, alpha2beta1, and hTERT. When cultured in serum- and androgen free medium, NHP cells gradually lose the expression of these markers, slow down in proliferation, and enter senescence. Several pieces of evidence implicate 15 lipoxygenase 2 (15-LOX2), a molecule with a restricted tissue expression and most abundantly expressed in adult human prostate, in the replicative senescence of NHP cells. First, the 15-LOX2 promoter activity and the mRNA and protein levels of 15-LOX2 and its multiple splice variants are upregulated in serially passaged NHP cells, which precede replicative senescence and occur in a cell-autonomous manner. Second, all immortalized prostate epithelial cells and prostate cancer cells do not express 15-LOX2. Third, PCa cells stably transfected with 15-LOX2 or 15-LOX2sv-b, a splice variant that does not possess arachidonate-metabolizing activity, show a passage-related senescence-like phenotype. Fourth, infection of early-passage NHP cells with retroviral vectors encoding 15-LOX2 or 15-LOX2sv-b induces partial cell-cycle arrest and big and flat senescence-like phenotype. Finally, 15-LOX2 protein expression in human prostate correlates with age. Together, these data suggest that 15-LOX2 may represent an endogenous prostate senescence gene and its tumor-suppressing functions might be associated with its ability to induce cell senescence. PMID- 15750633 TI - p53 must be competent for transcriptional regulation to suppress tumor formation. AB - In vitro studies suggest that effective tumor suppression by p53 requires multiple domains to execute transcription-dependent and transcription-independent functions. We generated a mutant p53 allele in mice, p53(W25QL26S) (p53(QS)), containing an inactive transactivation domain to evaluate the importance of transactivation for p53-mediated tumor suppression. Recently, we discovered that the allele also contains a valine substitution for alanine at codon 135, which borders the DNA-binding domain. We found that p53(QSval135) bound to chromatin albeit less well than p53(QSala135), but both were equally deficient in transcriptional regulation, apoptosis induction in mouse embryo fibroblasts (MEFs), and suppression of tumor formation by E1A, Ha-Ras transformed MEFs. p53(QSval135) mice and p53-null mice exhibited identical tumor development kinetics and spectra in spontaneous and oncogene-initiated tumorigenicity assays, when tested in a homo- and heterozygous configuration. The p53(QSval135) allele did not have dominant negative functions and behaved as a null allele. Taken together, these data indicate that effective tumor suppression requires the transcriptional regulation function of p53, and they suggest that transactivation independent functions of p53 are unlikely to contribute significantly to tumor suppression in vivo. PMID- 15750632 TI - YB-1 facilitates basal and 5-fluorouracil-inducible expression of the human major vault protein (MVP) gene. AB - Vaults have been suggested to play a direct role in multidrug resistance (MDR) to anticancer drugs. The human major vault protein (MVP) also known as lung resistance-related protein (LRP) represents the predominant component of vaults that may be involved in the defense against xenobiotics. Here, we demonstrate that besides MDR-related cytostatics, also the non-MDR-related drug 5 fluorouracil (5-FU) was able to induce MVP mRNA and protein expression. Treatment with 5-FU amplified the binding activity and interaction of the transcription factor Y-box binding protein-1 (YB-1) with the Y-box of the human MVP gene promoter in a time-dependent manner. 5-FU also induced reporter expressions driven by a panel of newly generated MVP promoter deletion mutants. Interestingly, stably YB-1 overexpressing cell clones showed enhanced binding of YB-1 to the Y-box motif, associated with enhanced basal as well as 5-FU-inducible MVP promoter-driven reporter expressions. Moreover, transduction of YB-1 cDNA led to increased expression of endogenous MVP protein. Under physiological conditions, we observed a strong coexpression of MVP and YB-1 in human colon carcinoma specimen. In summary, our data demonstrate a direct involvement of YB-1 in controlling basal and 5-FU-induced MVP promoter activity. Therefore, YB-1 is directly linked to MVP-mediated drug resistance. PMID- 15750634 TI - Bacterial death comes full circle: targeting plasmid replication in drug resistant bacteria. AB - It is now common for bacterial infections to resist the preferred antibiotic treatment. In particular, hospital-acquired infections that are refractory to multiple antibiotics and ultimately result in death of the patient are prevalent. Many of the bacteria causing these infections have become resistant to antibiotics through the process of lateral gene transfer, with the newly acquired genes encoding a variety of resistance-mediating proteins. These foreign genes often enter the bacteria on plasmids, which are small, circular, extrachromosomal pieces of DNA. This plasmid-encoded resistance has been observed for virtually all classes of antibiotics and in a wide variety of Gram-positive and Gram negative organisms; many antibiotics are no longer effective due to such plasmid encoded resistance. The systematic removal of these resistance-mediating plasmids from the bacteria would re-sensitize bacteria to standard antibiotics. As such, plasmids offer novel targets that have heretofore been unexploited clinically. This Perspective details the role of plasmids in multi-drug resistant bacteria, the mechanisms used by plasmids to control their replication, and the potential for small molecules to disrupt plasmid replication and re-sensitize bacteria to antibiotics. An emphasis is placed on plasmid replication that is mediated by small counter-transcript RNAs, and the "plasmid addiction" systems that employ toxins and antitoxins. PMID- 15750635 TI - Modified Mg : Al hydrotalcite in the synthesis of oxazolidin-2-ones. AB - The modified Mg : Al (3 : 1) hydrotalcite has been found to be an efficient catalyst in the conversion of carbamates into oxazolidin-2-ones under mild reaction conditions. A wide variety of oxazolidin-2-ones were obtained with excellent chemical yield. PMID- 15750636 TI - A multipolymer system for organocatalytic alcohol oxidation. AB - A system involving two polymer-supported reagents for the selective and organocatalytic oxidation of alcohols to aldehydes or ketones has been developed in which both polymeric reagents can be recovered and reused. PMID- 15750637 TI - Structural manifestations of the cheletropic reaction. AB - Examination of selected cyclopentenone derivatives which are fixed into the boat conformation reveals structural deviations from "normal" C-C(O) and C=O bond distances consistent with the early stages of the cheletropic extrusion of carbon monoxide. PMID- 15750638 TI - Asymmetric organotellurides as potent antioxidants and building blocks of protein conjugates. AB - New asymmetric organotellurides exhibiting good antioxidant properties in vitro and in cell culture can be attached to human serum albumin. PMID- 15750640 TI - Dye-functionalized head-to-tail coupled oligo(3-hexylthiophenes)-perylene oligothiophene dyads for photovoltaic applications. AB - A series of novel donor-acceptor systems, consisting of head-to-tail coupled oligo(3-hexylthiophene)s covalently linked to perylenemonoimide, is described. These hybrid molecules, which differ by the length of the oligothiophene units from a monothiophene up to an octathiophene, were created via effective palladium catalyzed Negishi and Suzuki cross-coupling reactions in good to excellent yields. The optical and electrochemical properties of these compounds were determined and based on this series structure-property relationships have been established which give vital information for the fabrication of photovoltaic devices. Because the synthesized perylenyl-oligothiophenes distinguish themselves by a high absorption between 300 and 550 nm and an almost complete fluorescence quenching of the perylene acceptor, they meet the requirements for organic solar cells. PMID- 15750639 TI - Kinetic resolution of sec-alcohols using a new class of readily assembled (S) proline-derived 4-(pyrrolidino)-pyridine analogues. AB - We report the development of a new class of readily prepared chiral 4 (pyrrolidino)-pyridine catalysts capable of exploiting both van der Waals (pi) and H-bonding interactions, thus allowing remote chiral information to stereochemically control the kinetic resolution of sec-alcohols. PMID- 15750641 TI - Synthesis of 2-pyridone-fused 2,2'-bipyridine derivatives. An unexpectedly complex solid state structure of 3,6-dimethyl-9H-4,5,9-triazaphenanthren-10-one. AB - 2-Pyridone-fused 2,2'-bipyridine derivatives and were synthesised. X-Ray diffraction analysis of revealed a highly complex solid state structure with a disordered molecule imbedded in a channel structure formed by a centrosymmetric lattice of hexagonally packed, hydrogen bonded columns. The columns are assembled from three symmetry independent molecules. Dimerisation of the self-complementary cis-amide hydrogen bond motif is overridden by the fulfilment of the proton coordination ability of the phenanthroline nitrogens in accordance with Etter's rules of hydrogen bond priorities. PMID- 15750642 TI - Locked TASC probes for homogeneous sensing of nucleic acids and imaging of fixed E. coli cells. AB - We have designed a second-generation TASC (target-assisted self-cleavage) probe. It is based on the switching-on of incorporated cis-acting DNAzyme activity upon the target-induced conformational change of the otherwise inactive off-target probes locked in an intrastrand base-paired hairpin geometry. With E. coli 16S ribosomal RNA-relevant oligonucleotides as targets, the locked TASC probe exhibits an allosteric factor of k(on)/k(off) = 65 and the sequence selectivity is high, in terms of single nucleotide difference, when particular sequence and length of targets are chosen. Preliminary experiments with fixed E. coli cells show that the locked TASC probe with a FRET pair can be used to image fixed E. coli cells. PMID- 15750643 TI - Effect of cyclodextrin complexation in bromine addition to unsymmetrical olefins: evidence for participation of cyclodextrin hydroxyl groups. AB - Multiple recognition by cyclodextrin in a bimolecular reaction, namely bromination of styrene, methyl cinnamate, phenylacetylene and allylbenzene, has been studied. Bromohydrin is obtained as a major product along with dibromide in the bromination of styrene and methyl cinnamate. The percentage of bromohydrin decreases as the cavity size increases. With phenylacetylene, bromophenylacetylene and phenacyl bromide are obtained in addition to the dibromides. In the bromination of cyclodextrin complexes of allylbenzene, the product distribution is the same as in solution bromination. The observed results demonstrate the efficiency of cyclodextrin in stabilizing the open carbocationic intermediate and thus provide chemical evidence for the participation of cyclodextrin hydroxyl groups. PMID- 15750644 TI - Synthesis and characterisation of highly emissive and kinetically stable lanthanide complexes suitable for usage "in cellulo". AB - The synthesis and photophysical characterisation are reported of a series of cationic, neutral and anionic europium and terbium complexes based on structurally related, nonadentate ligands based on the cyclen macrocycle. Each complex incorporates a tetraazatriphenylene moiety and overall absolute emission quantum yields are in the range 15-40% in aerated aqueous media. Dynamic quenching of the lanthanide excited state occurs with electron-rich donors, e.g. iodide, ascorbate and urate, and a mechanistic interpretation is put forward involving an electron transfer process. The cationic lanthanide complexes are taken up by NlH/3T3 cells and tend to localise inside the cell nucleus. PMID- 15750645 TI - Synthesis and antimycobacterial activity of agelasine E and analogs. AB - Agelasine E, previously isolated from the marine sponge Agelas nakamurai, has been synthesized for the first time, together with analogs with various terpenoid side chains. Treatment of N6-methoxy-9-methyl-9H-purin-6-amine with allylic bromides gave the desired 7,9-dialkylpurinium salts together with minor amounts of the N6-alkylated isomer. The N6-methoxy group was finally removed reductively. 1H-15N HMBC and 1H-15N HSQC NMR spectroscopy gave additional information on tautomerism and charge delocalization in the purine derivatives studied. The heterocyclic products were screened for activity against Mycobacterium tuberculosis and agelasine analogs carrying a relatively long terpenoid substituent in the purine 7-position and a methoxy group at N-6 were potent inhibitors of bacterial growth. Since agelasine analogs with the geranylgeranyl chain at N-7 exhibited antimicrobial activity, several strategies for synthesis of geometrically pure (2E,6E,10E)-geranylgeranyl bromide from geranyllinalool were evaluated. PMID- 15750646 TI - Triflic acid-catalyzed adamantylation of aromatics in [BMIM][OTf] ionic liquid; synthetic scope and mechanistic insight. AB - A mild and efficient process has been developed for the one-pot adamantylation of aromatic substrates employing 1-AdaOH, 1-AdaCl, 1-AdaBr, and 1-Br-3,5,7 trimethyladamantane as adamantylating agents, with triflic acid (TfOH) as promoter and n-butylmethylimidazolium triflate [BMIM][OTf] room temperature ionic liquid (IL) as solvent. The influence of reaction temperature, reaction time and the amount of TfOH was gauged in model reactions employing 1-AdaOH, 1-AdaCl and 1 AdaBr with toluene as the substrate. Under optimal conditions, the reactions exhibit high para selectivity with little or no adamantane side-product being formed. The synthetic scope of this transformation was tested for representative alkylbenzenes and haloalkylbenzenes. Comparative reactions carried out in 1,2 dichloroethane (DCE) produce increased amounts of the meta isomer and substantial amounts of adamantane. Substrate selectivities (K(T)/K(B)) were measured in competitive experiments in [BMIM][OTf] and in DCE as solvents. Isomerization tests were performed to shed light on the origin of the meta isomer. A DFT study was also conducted to compare relative stabilities of the isomeric products, to gauge the relative stabilities of the intermediate isomeric benzenium ions of adamantylation and their charge distribution modes, and to explore the intramolecular process for isomerization in the benzenium ion. PMID- 15750647 TI - 1-Oxabicyclic beta-lactams as new inhibitors of elongating MPT--a key enzyme responsible for assembly of cell-surface phosphoglycans of Leishmania parasite. AB - New iminosugars (1-oxabicyclic beta-lactam disaccharides) have been synthesized as inhibitors of elongating alpha-D-mannosyl phosphate transferase (eMPT), a key enzyme involved in the iterative biosynthesis of cell-surface phosphoglycans of the Leishmania parasite. The design is based on a transition-state model for this remarkable enzyme that transfers intact alpha-D-mannosyl-phosphate from GDP-Man. Since these phosphoglycans are unique to Leishmania and are essential for its infectivity and survival, their biosynthetic pathway has emerged as a novel target for anti-leishmanial drug and vaccine design. PMID- 15750648 TI - The facile preparation of primary and secondary amines via an improved Fukuyama Mitsunobu procedure. Application to the synthesis of a lung-targeted gene delivery agent. AB - An efficient modification of the Fukuyama-Mitsunobu procedure has been developed whereby primary or secondary amines can be synthesized from alkyl alcohols and the corresponding nosyl-protected/activated amine. Most importantly, the use of the DTBAD and diphenylpyridinylphosphine, as Mitsunobu reagents, generates reaction by-products that can be easily removed, providing a remarkably clean product mixture. This improved technique was implemented in the synthesis of a complex lipopeptide designed to target alpha9beta1-integrin proteins predominant on upper airway epithelial cells. PMID- 15750649 TI - Side chain homologation of alanyl peptide nucleic acids: pairing selectivity and stacking. AB - Alanyl peptide nucleic acids (alanyl-PNAs) are oligomers based on a regular peptide backbone with alternating configuration of the amino acids. All side chains are modified by covalently linked nucleobases. Alanyl-PNAs form very rigid, well defined, and linear double strands based on hydrogen bonding of complementary strands, stacking, and solvation. Side chain homology was examined by comparing a methylene linker (alanyl-PNA) with an ethylene linker (homoalanyl PNA), a trimethylene linker (norvalyl-PNA), and PNA sequences with mixed linker length between nucleobase and backbone. Side chain homology in combination with a linear double strand topology turned out to be valuable in order to selectively manipulate pairing selectivity (pairing mode) and base pair stacking. PMID- 15750650 TI - Easy synthesis of beta-O-4 type lignin related polymers. AB - The beta-O-4 structure is the most abundant substructure in lignin. Lignin related polymers composed of only the beta-O-4 structure were prepared using simple aromatic compounds as starting materials. Acetophenone derivatives were brominated, polymerized in the presence of K2CO3 and reduced with NaBH4 to give the lignin related polymers. These are linear polymers which resemble natural lignins in their structures, although they do not have a gamma-hydroxymethyl group. The number average degree of polymerization (DPn) was determined with peracetate of the polymers by gel permeation chromatography. The DPn of guaiacyl type polymers ranged from 15.2-21.4, where the value for the syringyl type was 11.3 and for the p-hydroxyphenyl type 16.9. The Guaiacyl type polymer was very soluble in usual lignin solvents such as 1, 4-dioxane-water (96 : 4, v/v) and DMSO, but only slightly soluble in acetone-water (9 : 1, v/v). PMID- 15750651 TI - Novel carbamate derivatives of 4-beta-amino-4'-O-demethyl-4-desoxypodophyllotoxin as inhibitors of topoisomerase II: synthesis and biological evaluation. AB - A novel series of carbamate derivatives of 4-beta-amino-4'-O-demethyl-4 desoxypodophyllotoxin were synthesized. Their effect on human DNA topoisomerase II and antiproliferative activity was evaluated. Compounds 4a-c, 4g, 4j and 4k are topoisomerase II poisons that induce double-stranded breaks in DNA and exhibit increased cytotoxicity compared to etoposide. PMID- 15750652 TI - A chemoenzymatic total synthesis of the undecenolide (-)-cladospolide B via a mid stage ring-closing metathesis and a late-stage photo-rearrangement of the E isomer. AB - A sixteen-step synthesis of the twelve-membered macrolide (-)-cladospolide B(2) from the microbially-derived cis-1,2-dihydrocatechol 5 is described. Pivotal steps include the ring-closing metathesis (RCM) of diene 12 to give the ten membered lactone 13 together with small amounts of the head-to-tail and head-to head dimers 14 and 15, respectively. The saturated lactol 19 derived from compounds 13 and 14 readily participates in a Wadsworth-Horner-Emmons reaction to give the E-configured alpha,beta-unsaturated ester 20. This last compound is then converted, through application of a Yamaguchi lactonisation reaction on the derived acid 22, into the macrolide 23 which, upon removal of the bis-acetal protecting group, affords compound 24, the E-isomer of target 2. Irradiation of a benzene solution of compound 24 results in its partial photoisomerisation to (-) cladospolide B(2). PMID- 15750653 TI - Hydrolytic stability of 2',3'-O-methyleneadenos-5'-yl 2',5'-di-O-methylurid-3'-yl 5'-O-methylurid-3'(2')-yl phosphate: implications to feasibility of existence of phosphate-branched RNA under physiological conditions. AB - Hydrolytic reactions of 2',3'-O-methyleneadenos-5'-yl 2',5'-di-O-methylurid-3'-yl 5'-O-methylurid-3'(2')-yl phosphate (1a,b) have been followed by RP-HPLC over a wide pH range to evaluate the feasibility of occurrence of phosphate-branched RNA under physiological conditions. At pH <2, where the decomposition of is first order in [H3O+], the P-O5' bond is cleaved 1.5 times as rapidly as the P-O3' bond. Under these conditions, the reaction probably proceeds by an attack of the 2'-OH on the phosphotriester monocation. Over a relatively wide range from pH 2 to 5, the hydrolysis is pH-independent, referring to rapid initial deprotonation of the attacking 2'-OH followed by general acid catalyzed departure of the leaving nucleoside. The P-O5' bond is cleaved 3 times as rapidly as the P-O3' bond. At pH 6, the reaction becomes first order in [HO-], consistent with an attack of the 2'-oxyanion on neutral phosphate. The product distribution is gradually inversed: in 10 mmol L(-1) aqueous sodium hydroxide, cleavage of the P O3' bond is favored over P-O5' by a factor of 7.3. The results of the present study suggest that the half-life for the cleavage of under physiological conditions is only 100 s. Even at pH 2, where is most stable, the half-life for its cleavage is less than one hour and the isomerization between and is even more rapid than cleavage. The mechanisms of the partial reactions are discussed. PMID- 15750654 TI - The mechanistic origin of regiochemical changes in the nitrosative N-dealkylation of N,N-dialkyl aromatic amines. AB - The regioselectivity of the nitrous acid mediated dealkylation of 4-substituted-N ethyl-N-methylanilines is a function of the acidity of the reaction mixture. At high acidity deethylation predominates, whereas demethylation is the predominant reaction in nitrosamine formation at pH 2 and above. In some cases the regioselectivity of nitrosative dealkylation changes as the run proceeds. Through the use of the corresponding 4-nitroaniline as the primary substrate, CIDNP, kinetics, kinetic deuterium isotope effects and other transformations involving nitrosations with NO2 or NOBF4 in aprotic solvents, a new mechanism of tertiary amine nitrosation has been deduced and proposed to explain regioselective deethylation. The mechanism involves the oxidation of the substrate to the amine radical cation by NO+. This is followed by the abstraction of a hydrogen atom from the carbon adjacent to the amine nitrogen by NO2 to produce an iminium ion which reacts further to produce the corresponding aldehyde and the nitrosamine. Depending upon the acidity, this process competes with three other mechanistic pathways, two of which give the nitrosamine through the iminium ion, and one leads to the formation of C-nitro compounds. The competing pathways to nitrosamine formation involve NOH elimination from a nitrosammonium ion and deprotonation of the radical cation to give an alpha-amino radical which rapidly oxidized to the iminium ion. Predominant, but not highly regioselective demethylation occurs by these pathways. Nitro compound formation principally arises from the reaction of NO2 with the radical cation followed by deprotonation, but also occurs by para C-nitrosation followed by oxidation. PMID- 15750655 TI - Synthesis and evaluation of mimetics of UDP and UDP-alpha-D-galactose, dTDP and dTDP-alpha-D-glucose with monosaccharides replacing the key pyrophosphate unit. AB - A series of 5'-O-glycosyl-uridine and thymidine derivatives have been prepared as potential mimics of sugar nucleotides and nucleotide-diphosphates. These compounds proved not to be inhibitors of bovine beta-1,4-galactosyltransferase although some showed moderate inhibition of Salmonella dTDP-alpha-D-glucose 4,6 dehydratase (RmlB). PMID- 15750656 TI - Enantiomeric cannabidiol derivatives: synthesis and binding to cannabinoid receptors. AB - (-)-Cannabidiol (CBD) is a major, non psychotropic constituent of cannabis. It has been shown to cause numerous physiological effects of therapeutic importance. We have reported that CBD derivatives in both enantiomeric series are of pharmaceutical interest. Here we describe the syntheses of the major CBD metabolites, (-)-7-hydroxy-CBD and (-)-CBD-7-oic acid and their dimethylheptyl (DMH) homologs, as well as of the corresponding compounds in the enantiomeric (+) CBD series. The starting materials were the respective CBD enantiomers and their DMH homologs. The binding of these compounds to the CB(1) and CB(2) cannabinoid receptors are compared. Surprisingly, contrary to the compounds in the (-) series, which do not bind to the receptors, most of the derivatives in the (+) series bind to the CB(1) receptor in the low nanomole range. Some of these compounds also bind weakly to the CB(2) receptor. PMID- 15750657 TI - Zinc-mediated carbon radical addition to glyoxylic imines in aqueous media for the synthesis of alpha-amino acids. AB - The addition of carbon radicals to glyoxylic imines was studied using zinc dust as a radical initiator. The zinc-mediated radical reaction of glyoxylic oxime ethers and hydrazones proceeded smoothly to give the alkylated products via a carbon-carbon bond-forming process in aqueous media. The reaction of the oxime ethers and hydrazones having an Oppolzer's camphorsultam group provided the corresponding alkylated products, which could be converted into enantiomerically pure alpha-amino acids. The diastereoselectivities observed in the reaction of hydrazones were better than those obtained in the reaction of oxime ethers. PMID- 15750658 TI - Evidence of a self-inclusion phenomenon for a new class of mono-substituted alkylammonium-beta-cyclodextrins. AB - A new class of mono-substituted N-alkyl-N,N-dimethylammonium-beta-cyclodextrins has been synthesized in a three step procedure from the native beta-cyclodextrin. The structural analysis of these compounds undertaken by combined use of 1D and 2D NMR spectra indicate that the two methyl groups bound on the nitrogen are magnetically inequivalent due to a self-inclusion phenomenon of the alkyl chain inside the CD cavity. A variable-temperature 1H NMR study showed that these mono substituted CD derivatives formed temperature-independent intramolecular complexes with their own alkylammonium substituent. The strength of the interaction between the alkyl moiety and the cyclodextrin cavity has been evaluated by a competitive method using an adamantane derivative. Finally, surface tension measurements demonstrated the surface active character of these compounds and confirmed their self-inclusion ability. PMID- 15750659 TI - Synthesis of unsymmetrical 3,3'-biquinazoline-2,2'-diones by condensation of 3 aminoquinazolinones with benzoxazinones; fortuitous discovery, and further syntheses of 4-H-3-oxo-1,9a,10-triazaanthracen-9-ones. AB - Condensation of 2-alkyl- or 2-aryl-3-aminoquinazolin-4-ones with benz[1,3]oxazin 4-ones gives the unsymmetrical 2,2' disubstituted 3,3' biquinazoline-4,4'-diones. The reaction is tolerant to a range of heteroatom and unsaturated functionality in the quinazolinone 2-position. However, treatment of 3-amino-2-hydroxymethyl-3H quinazolin-4-ones with benz[1,3]oxazinone at high temperatures gave 4H-3-oxo 1,9a,10-triazaanthracen-9-ones, an unreported fused heterocyclic system, a more direct synthesis of which, replacing benzoxazinones with orthoesters is presented. PMID- 15750660 TI - Do protein and energy intakes explain long-term changes in body composition? AB - BACKGROUND: Despite evidence that profiles of protein and energy intake can determine short-term (< 1 y) change in both lean and fat compartments,the role of diet in longer-term, age-related changes in body composition remains unclear. OBJECTIVE: This paper tests for long-term counterparts to the well-established short-term relationships between protein and energy intake and changes in body composition. DESIGN: Using longitudinal data on 608 healthy, non-obese Chinese (50-69 y)from the 1993 and 1997 China Health and Nutrition Surveys, sex-specific regression models were created to determine if 3-day mean protein (%of energy) and energy (kJ) intakes at baseline predicted change in mid arm muscle area (MAMA) and waist circumference (WC). RESULTS: Although sex-specific U-shaped associations were observed,higher energy intakes were associated with greater gain in WC and less loss of MAMA, and higher protein intakes with less loss of MAMA than lower intakes for both sexes, adjusting for baseline age, height, weight, MAMA, WC, smoking status, activity level, income and urban residence. For males, energy intake be low 95%of the Chinese RDA was associated with significantly smaller gains in WC and greater loss of MAMA than energy intake between 95-125% RDA. For both sexes, protein intake be low 10.4% of energy was associated with significantly greater loss of MAMA than intake between 10.4-12.1% of energy. For females, energy intake above 125% RDA was associated with significantly greater gains in body fat than intake between 95-125% RDA. CONCLUSION: The results suggest that diet may play an important role in age related change in body composition. PMID- 15750661 TI - Percent body fat estimation from skin fold thickness in the elderly. Development of a population-based prediction equation and comparison with published equations in 75-year-olds. AB - OBJECTIVE: To develop a prediction equation for percent body fat (BF%) from skin fold thickness in a random sample of free-living 75-year-olds and to compare with published equations for the elderly. SETTING: Goteborg part of the Nordic Research on Aging (NORA) study in 1991/92. SUBJECTS & METHODS: Anthropometric measurements were performed in 286 participants (125 males,161 females) and body composition was estimated by four compartment model (4C) from total body water (TBW) and total body potassium (TBK) in every alternative subjects. BF % predicted from skin fold thickness (BFSF) was validated against BF % estimated from 4C model (BF4C) and was compared with BFSF and BF % predicted from equations by Durnin and Womersley, 1974 (BFDW), Deurenberg et al.,1989 (BFPD) and, Visseretal.,1994(BFMV), respectively. RESULTS: The BFSF correlated well with BF4C (r = 0.86, SEE = 4.05). Compared to BF4C, both BFPD and BFMV over estimated BF % by 3.26 and 3.63 in males and 9.56 and 9.23 in females, respectively. BFDW underestimated BF % in females by 1.06 and overestimated by 3.12 in males. BFSF showed best agreement with BF4C, where the 95 % of differences lie between +/- 7.4 in males and +/- 8.7 in females, respectively. CONCLUSION: BF % predicted from skin fold thickness correlated well with estimated BF % in 75-year-olds. Different prediction equations gave different values for BF % and population specific prediction equations seem preferable in the elderly population. PMID- 15750663 TI - Dietary omega-3 Fatty acids and psychiatry: mood, behaviour, stress, depression, dementia and aging. AB - In view of the high omega-3 poly unsaturated fatty acid content of the brain, it is evident that these fats are involved in brain biochemistry, physiology and functioning; and thus in some neuropsychiatric diseases and in the cognitive decline of ageing. Though omega-3 fatty acids (from fatty fish in the human diet) appear effective in the prevention of stress, their role as regulator of mood and of libido is a matter for discussion pending experimental proof in animal and human models. Dietary omega-3 fatty acids play a role in the prevention of some disorders including depression, as well as in dementia, particularly Alzheimer's disease. Their direct role in major depression, bipolar disorder (manic depressive disease) and schizophrenia is not yet established. Their deficiency can prevent the renewal of membranes, and thus accelerate cerebral ageing; none the less, the respective roles of the vascular component on one hand (where the omega-3's are active) and the cerebral parenchyma itself on the other, have not yet been clearly resolved. The role of omega-3 in certain diseases such as dyslexia and autism is suggested. In fact, omega-3 fatty acids participated in the first coherent experimental demonstration of the effect of dietary substances (nutrients) on the structure and function of the brain. Experiments were first of all carried out one x-vivo cultured brain cells (1), then on in vivo brain cells(2), finally on physiochemical, biochemical, physiological, neurosensory, and behavioural parameters (3). These findings indicated that the nature of poly unsaturated fatty acids(in particular omega-3) present in formula milks for infants (both premature and term) determines the visual, cerebral,and intellectual abilities, as described in a recent review (4). Indeed,the insufficient dietary supply of omega-3 fatty acids in today's French and occidental diet raises the problem of how to correct dietary habits so that the consumer will select foods that are genuinely rich in omega-3/ the omega-3 family ; mainly rapeseed, (canola) and walnut oils on one hand and fatty fish on the other. PMID- 15750662 TI - Does eating environment have an effect on food intake in the elderly? AB - Identical meals were prepared and served to 49 elderly subjects (mean age 74.3 +/ SD 7.7 years) in two different eating environments. The improved environment was a state of the art training restaurant and the standard environment was a staff canteen. Each subject was served an identical meal in both environments. The energy intake of each meal consumed was determined using the weighed intake method. There was a significant difference (p < 0.001) in the energy intake in the elderly between the improved (4894 +/- 613 kJ) and standard (4536 +/- 620 kJ) eating environments. Eating environment has a positive effect on the energy intake of elderly subjects. Changes to eating environment may be a useful way to improve energy intake in the elderly. PMID- 15750664 TI - Low erythrocyte folate, but not plasma vitamin B-12 or homocysteine,is associated with dementia in elderly Latinos. AB - The relationship between B vitamin status and cognitive function has been of interest for many years. There is evidence of relationships between intake and status of folate and vitamin B-12 with neurological, cognitive, and memory impairment, but results have been inconsistent. Plasma B-12, erythrocyte folate, methylmalonic acid,and homocysteine were evaluated as predictors of cognitive function in a large population based sample of Latino elderly living in the Sacramento, California region. The hypothesis tested was that low folate and/or B 12 status predicts cognitive function impairment and dementia. Logistic regression was used to examine the differences in B-vitamin status by cognitive function category. Erythrocyte folate was related to dementia after controlling for age, gender, education, income, diabetes diagnosis, serum creatinine, and depressive symptoms. The highest prevalence of low erythrocyte folate occurred in the Dementia group and was significantly higher than in the Normal group. Plasma B-12, MMA, Hcy, and prevalence of a normal values for these variables, were not significantly different among the cognitive function classes. We conclude that folate status is associated with dementia but that more research is needed on the relationship between vitaminB-12 status, Hcy and cognitive function to explore possible associations with these parameters. PMID- 15750665 TI - Zinc in health and chronic disease. AB - Zinc is a trace element essential for the optimal function of a variety of biochemical and physiological processes. Its role in healthy aging is particularly important as it prevents neo plastic cell growth, is involved in mitotic cell division, DNA and RNA repair. Although zinc is widely available in food, the daily intake in many persons may be suboptimal. Other causes of low zinc concentrations may be due to small bowel conditions that cause mucosal damage and thus decrease absorption. Chronic diseases associated with alterations in zinc status are bronchial asthma, rheumatoid arthritis and Alzheimer disease. At present it is uncertain if therapy with zinc would assist in the management of these chronic diseases. In view of the important cellular functions of zinc in the human body, a diet with an adequate zinc content is beneficial in promoting healthy aging and maintaining good health. PMID- 15750666 TI - A comparison of first event coronary heart disease rates in two contrasting California populations. AB - OBJECTIVES: To directly evaluate whether a population, many of whom are vegetarian or near vegetarian, does experience less coronary heart disease (CHD). METHODS: Two concurrent California observational studies,one with unusual dietary habits, are compared. Similar diagnostic criteria were used in both the Adventist Health Study and the Stanford Five-City Project. RESULTS: Age-standardized rate ratios (Adventist/Stanford study) for first event fatal CHD were 0.59 (95% confidence interval,0.43-0.80) in men and 0.49(0.32-0.76) in women. Similar ratios for first event myocardial infarction were 0.60(0.47-0.78) and 0.46 (0.33 0.65). Ratios are usually much lower at younger ages or when comparisons include only vegetarian Adventists. CONCLUSION: Lifestyle factors can markedly change the first-event CHD experience of whole populations, and appear to especially protect against premature events. PMID- 15750667 TI - Elevation of plasma homocysteine in natural menopause can not be explained by a lack of vitamin coenzyme availability: relevance to the risk of cardiovascular disease. AB - The risk of cardio vascular disease (CVD) doubles after menopause. Plasma homocysteine (hCy) is a risk factor which is influenced by vitamins B12,B6 and folate. The present study was conducted to examine the relationship of plasma hCy to the three vitamins and other contributing variables in early natural menopause. METHODS: Participants were healthy, non smoking Caucasian women 3 to 5 years postmenopausal (n = 26) or premenopausal between 30 and 45 y(n = 30). Anthropometric data, dietary records and plasma concentrations of hCy, vitamin B6, vitamin B12 and folate were obtained. RESULTS: The nutritional status of vitamins B6, B12 and folate as measured by dietary intake and blood concentrations was adequate in both groups. Mean fasting plasma total (t) hCy concentration of postmenopausal group was 2-fold higher than the value found for control group (P < 0.0001) without oral methionine loading. The difference between the two groups remained highly significant after adjustment for confounding variables by multivariate analysis, suggesting that the effect of estrogen deficiency was direct. CONCLUSION: In addition to the loss of the protective effects of estrogen on their cardiovascular physiology and lipid metabolism, postmenopausal women are exposed to higher plasma hCy concentrations and deleterious cardiovascular effects. The exact mechanism is not known but does not seem to be related to coenzyme deficiency. PMID- 15750668 TI - [Extracranial meningioma. Rare differential diagnosis of a subacute tumor]. PMID- 15750669 TI - [HALS: HIV-Lipodystrophy associated syndrome]. PMID- 15750671 TI - [Chronic graft-versus-host disease]. PMID- 15750670 TI - [Granuloma development after wrinkle injection with polylactate]. PMID- 15750672 TI - [Blue-rubber-bleb-nevus syndrome]. PMID- 15750673 TI - [Topical treatment of subacute-cutaneous lupus erythematosus with tacrolimus]. PMID- 15750674 TI - [Eccrine porocarcinoma of the head. Report 3 cases]. PMID- 15750675 TI - [Botulinum toxin treatment of eccrine sweat gland nevus]. PMID- 15750676 TI - [Photodynamic therapy for a nevoid basal cell carcinoma syndrome]. PMID- 15750677 TI - [Topical treatment of skin aging]. AB - Safe levels of UV exposure and UV protection are the most important measures to protect the skin from epithelial skin cancer and skin aging. This report reviews noninvasive topical methods to counteract skin wrinkling and irregular pigmentation of aging skin. Furthermore, information is provided about the effects of UV protection by using sunscreens and topical antioxidants. The effect of vitamin A acid derivatives, chemical peeling, and bleaching agents is considered. Newly developed substances are presented. PMID- 15750678 TI - [Local recurrence following radical cystectomy for bladder cancer. Diagnostic and therapeutic options]. AB - The prognosis for patients with local recurrence following cystectomy for urothelial bladder cancer is poor. Only a small proportion of patients with good performance status are candidates for any form of therapy at all. Clinical experience shows that local recurrence is often accompanied or followed by systemic tumor spread. Therefore, palliative systemic chemotherapy is the cornerstone of treatment. Local radiotherapy or local tumor resection is reserved for subgroups of patients and to ease local symptoms or complications. Only a few patients are candidates for multimodal therapeutic approaches with curative intent. Despite such efforts, the survival of patients with local recurrence is limited in nearly all cases. PMID- 15750679 TI - [Recurrent disease in renal cell carcinoma. "Local recurrence" after kidney sparing and radical resection]. AB - Locally recurrent renal cell carcinoma (RCC) is 0-10% after nephron-sparing surgery, 2.5-4% after thermoablative interventions and 2-3% after (radical) nephrectomy. Risk-factors are: sporadic or hereditary origin, tumor size, multifocality, histologic phenotype and incomplete resection. To date, there are no significant differences in the incidence of locally recurrent tumors independently of whether open or laparoscopic techniques were preferred. Caution still has to be taken with the use of alternative tools for minimally invasive tumor ablation.Finally, no statistically proven standard therapy exists that would clearly provide a superior outcome for patients with an isolated local recurrence. However, meta-analyses strongly support the performance of a resection of the recurrence as the primary working principal. PMID- 15750680 TI - Outcome of late-onset rheumatoid arthritis. AB - The objective of this study was to determine possible differences in the outcome of patients with rheumatoid arthritis (RA) with disease onset early and late in life. As part of a broader outcome study of RA which included patients seen in the division of Rheumatology of Hospital Universitario Marques de Valdecilla of Santander, Cantabria (Northern Spain) with disease duration between 2 and 7 years, we selected patients with an age at disease onset of or=65 years. The medical records of all eligible patients were reviewed for relevant clinical and laboratory variables; the patients were then further evaluated for disease activity using biological tests and joint indices such as joint counts and Thompson's Index, functional capacity using the American College of Rheumatology (ACR) functional classification (ACR-FC) and the modified Health Assessment Questionnaire (M-HAQ), and anatomical damage using the number of joint damage (NJD) and radiographs read by the Sharp's scoring method for joint erosion (JE), joint narrowing (JN), and overall. Patients in both subsets were then compared. For the multivariable analyses all patients in the original larger cohort were included, so that age could be used as a continuous variable and the power of the analyses could increase; 31 younger (mean age+/-SD: 36+/-7 years) and 35 older (73+/-6 years) patients were available for assessment. No differences in disease duration and gender distribution were observed. Likewise, both subsets had similar levels of disease activity, both articular indices, and biological markers. In contrast, elderly patients showed more functional limitations as per the M-HAQ [median (interquartile range): 0.4 (0.13-1.2) vs 0.13 (0-0.6), p=0.007] and greater anatomical damage as per the NJD [median (interquartile range): 2 (0-4) and 0 (0-2), respectively, p=0.04] and the JE, JN, and total Sharp Index score (p=0.001, 0.02, and 0.001, respectively). Although older patients took fewer disease-modifying antirheumatic drugs (DMARD) and combined DMARD treatments (2.5+/-1.4 vs 1.9+/-1.3, p=0.05 and 0.8+/-1.1 vs 0.3+/ 0.6, p=0.01, respectively), multivariable analysis demonstrated an independent association between age at disease onset and the number of DMARD and functional and anatomical decline. Late-onset RA does not present a better prognosis than the early-onset form of the disease. At the very least the disease is comparable between both patient groups. However, disease compounded by age-associated factors may in fact have a worse prognosis late than early in life. PMID- 15750681 TI - Animal models for fracture treatment in osteoporosis. AB - Demographic changes in the age structure of occidental populations are giving rise to osteoporosis and associated fractures, which are becoming a major public health burden. Various animal models have been established and used to investigate the pathogenesis of osteoporosis and to facilitate the preclinical testing of new treatment options such as antiresorptive drugs. Although osteoporosis can be induced in animals, spontaneous fractures without adequate trauma were only found in nonhuman primates. An animal model designed to investigate new ways to treat fractures of osteoporotic bone has to fulfill requirements that are very different from those of pharmacological testing. The aspects of major interest in orthopedic applications are bone fragility, efficacy of implant fixation and bone healing. Existing animal models for osteoporosis were critically reviewed focusing on these aspects. The advantages and disadvantages of the models with regard to their application in the testing of new fracture-fixation devices or biological approaches to stimulate bone healing are discussed. Ovariectomy alone does not cause the bone loss seen in osteoporotic human patients. New models to simulate fracture of osteoporotic bone need to be explored and used to address the specific aims of an experiment. PMID- 15750683 TI - Sexuality in multiple sclerosis. AB - Sexuality and partnership have an important influence on the quality of life of every person and also on people with chronic disorders such as multiple sclerosis. The findings in literature show high evidence that people with multiple sclerosis experience high levels of sexual dysfunction, most of them with hypoactive sexual behaviour often associated with dissatisfaction in relationship, and also the partners seem to show lower sexual and partnership satisfaction. The most common problems in women are lack of sexual interest and decreased libido, often with problems in orgasmic capacity, while men report erectile dysfunction and also lack of sexual interest. The impact of the level of disability and duration of the illness remains unclear. Positive familial support can often help the patient in coping with the illness, nonetheless problems with changing roles and multiple-sclerosis-minimizing can improve the need of contacts to outstanding persons. PMID- 15750684 TI - Atomic force microscopy investigation of chemically stabilized pericardium tissue. AB - Native and chemically stabilized porcine pericardium tissue was imaged by the contact mode atomic force microscopy (AFM), in air. Chemically stabilized pericardium is used as a tissue-derived biomaterial in various fields of the reconstructive and replacement surgery. Collagen type I is the main component of the fibrous layer of the pericardium tissue. In this study, the surface topography of collagen fibrils in their native state in tissue and after chemical stabilization with different cross-linking reagents: glutaraldehyde (GA), dimethyl suberimidate (DMS) and tannic acid (TA) was investigated. It has been found that chemical stabilization causes considerable changes in the surface topography of collagen fibrils as well as in the spatial organization of the fibrils within the tissue. The observed changes in the D-spacing pattern of the collagen fibril correspond to the formation of intrafibrilar cross-links, whereas formation of interfibrilar cross-links is mainly responsible for the observed tangled spatial arrangement of fibrils and crimp structure of the tissue surface. The crimp structure was distinctly seen for the GA cross-linked tissue. Surface heterogeneity of the cross-linking process was observed for the DMS-stabilized tissue. SDS-PAGE electrophoresis was performed in order to evaluate the stabilization effect of the tissues treated with the cross-linking reagents. It has been found that stabilization with DMS, GA or TA enhances significantly the tissue resistance to SDS/NaCl extraction. The relation between the tissue stability and changes in the topography of the tissue surface was interpreted in terms of different nature of cross-links formed by DMS, GA and TA with collagen. PMID- 15750682 TI - N-PEP-12--a novel peptide compound that protects cortical neurons in culture against different age and disease associated lesions. AB - The neuroprotective potency of N-PEP-12, a novel, proprietary compound consisting of biopeptides and amino acids was investigated. Lesion models have been applied in neuronal cultures of embryonic chicken cortex, pre-treated with N-PEP-12 from the first day onwards. On day 8 in vitro neurons were lesioned and cell viability was measured 24 and 48 hours later. To simulate acute brain ischemia, cytotoxic hypoxia was induced by sodium cyanide or by iodoacetate and excitotoxicity by L glutamate. Ionomycin for up to 48 hours induced calcium overload. The cytoskeleton was disrupted by addition of colchicine. N-PEP-12 shows dose dependent neuroprotection in all different models. The effect size depends on the recovery time but also on the extent of the lesion. In cases of mild to moderate lesion pronounced dose-dependent effects could be demonstrated. This indicates that chronic exposure to N-PEP-12 is able to prevent neuronal cell death associated to conditions occurring during normal aging and neurological disorders like ischemic stroke, hypoxia, brain trauma, or AD. PMID- 15750685 TI - Post-zygotic de novo trinucleotide repeat expansion at spinocerebellar ataxia type 7 locus: evidence from an Indian family. AB - Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant cerebellar ataxia caused by CAG repeat expansion. We found expansion at SCA7 locus in only two out of 235 Indian families clinically diagnosed for ataxia. In one of the families, a de novo mutation was observed wherein a paternal allele in intermediate range of 31 CAG repeats expanded to 59 in the offspring leading to the disease. No expanded alleles were observed in the sperm of the transmitting parent by small pool PCR. This suggests that de novo expansion by a pre-zygotic event is unlikely and could be post-zygotic. SCA7 expanded alleles from the two families were present on different genetic backgrounds, indicating multiple origins of the mutation. PMID- 15750686 TI - Guidelines for the use of biochemical markers of bone turnover in osteoporosis (2004). PMID- 15750688 TI - Expression of osteopontin mRNA in odontoclasts revealed by in situ hybridization during experimental tooth movement in mice. PMID- 15750687 TI - Guidelines on the management and treatment of glucocorticoid-induced osteoporosis of the Japanese Society for Bone and Mineral Research (2004). PMID- 15750689 TI - Identification of CBFA1-regulated genes on SaOs-2 cells. AB - Current knowledge about mechanisms controlling osteoblast-specific gene expression has led to the identification of Cbfa1 as a key regulator of osteoblast differentiation. Several essential questions about this transcription factor remain to be addressed, e.g., the nature of stimuli that may modulate its own expression, as well as the genetic repercussions following alterations in Cbfa1 levels. To identify such Cbfa1-responsive genes, the SaOs-2 cell line was stably transfected with a dominant negative mutant of Cbfa1 (DeltaCbfa1). Comparison of gene expression patterns by differential display on selected SaOs-2 clones allowed the identification of four new genes that may be under the control of Cbfa1. Three of them, SelM, elF-4AI, and RPS24, seemed to be linked to a global change in cellular metabolism and cell growth. The fourth, the CD99/MIC2 gene, was strongly overexpressed (around tenfold) in cells presenting high levels of Deltacbfa1. This observation adds evidence to show that this marker of Ewing family tumors is linked to the osteoblast lineage. The exact function of CD99 remains largely undefined, and this is the first time that its regulation by an essential transcription factor involved in osteoblast differentiation has been observed. PMID- 15750690 TI - Molecular and cell biological properties of mouse osteogenic mesenchymal progenitor cells, Kusa. AB - A cell line of murine osteogenic progenitor cells, Kusa, was established from femoral bone marrow stromal cells with other types of mesenchymal progenitor cells. We characterized two sublines of Kusa (Kusa-A1 and Kusa-O) from several aspects, including the use of an expression profiling system, a cDNA microarray. The original Kusa subline (Kusa-A1) had high alkaline phosphatase activity and high accumulation of calcium deposits in a condition inducing mineralization, with ascorbic acid and beta-glycerophosphate. Kusa-O, a low osteogenic subline of Kusa, had high alkaline phosphatase activity but slow accumulation of calcium deposits even in the inducing condition. These two Kusa sublines differed in the expression of the osteogenic marker genes, osteocalcin and osteopontin, during mineralization. A type of cDNA microarray revealed marked downregulation of gene expression in the inducing condition in both Kusa-A1 and Kusa-O. Another type of high-throughput microarray was performed to examine the difference in gene expression patterns between Kusa-A1 and Kusa-O. By this analysis, periostin, which would be involved in a stage of osteogenesis, was low in Kusa-A1. On the contrary, Myocyte enhancer factor 2C (MEF2C), a myogenic transcriptional factor, was high in Kusa-A1, although no expression of any other myogenic genes was shown. PMID- 15750691 TI - Supplementation of L-arginine prevents glucocorticoid-induced reduction of bone growth and bone turnover abnormalities in a growing rat model. AB - The present study was designed to evaluate the effects of glucocorticoid (GC) treatment on bone turnover and bone mineral density in the growing rat. Because of the recent evidence that nitric oxide (NO) can counteract prednisolone-induced bone loss in mature rats, we examined the effect on bone of the NO donor L: arginine in young male rats, in which bone mass is increased by the same biological mechanism as in children and adolescents. Thirty-six 10-week-old Sprague-Dawley male rats were assigned to six groups of six animals each, and treated for 4 weeks with either vehicle (once a week subcutaneous injection of 100 microl of sesame oil); prednisolone sodium succinate, 5 mg/kg, 5 days per week by intramuscular injection (i.m.); L-arginine, 10 mg/kg intraperitoneally (i.p.) once a day; N(G)-nitro-L-arginine methylester (L-NAME), 50 mg/kg subcutaneously once a day; prednisolone sodium succinate 5 mg/kg, 5 days per week i.m. +L-arginine 10 mg/kg i.p. once a day; or prednisolone sodium succinate, 5 mg/kg, 5 days per week i.m. +L-NAME 50 mg/kg subcutaneously once a day. Serum calcium, alkaline phosphatase (ALP), osteocalcin, and the C-terminal telopeptides of type I collagen (RatLaps) were measured at baseline conditions and after 2 and 4 weeks. Prior to treatment, and after 2 and 4 weeks, the whole body, vertebral, pelvic, and femoral bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA) scanning. Prednisolone and prednisolone+L-NAME treated rats had significantly lower ALP and osteocalcin levels than controls at 2 and 4 weeks, and significantly higher levels of Rat-Laps than controls at 4 weeks. Prednisolone, L-NAME, and prednisolone+L-NAME produced a significant inhibition of bone accumulation and bone growth at all sites measured. Supplementation with L-arginine appeared to prevent the inhibition of bone growth and increase in bone resorption induced by prednisolone. These data would suggest, for the first time, that supplementation with an NO donor could be considered as a treatment for steroid-induced osteoporosis in the developing skeleton. PMID- 15750693 TI - Seasonal changes of serum 25-hydroxyvitamin D and intact parathyroid hormone levels in a normal Japanese population. AB - We conducted an observational study in order to assess the prevalence of hypovitaminosis D and its seasonal changes, in the Tokai area (N35.3 E137.0), in 197 normal subjects in Japan. The mean serum 25-hydroxyvitamin D (25-OHD) level measured by direct radioimmunoassay (RIA) was lowest at the end of winter, and highest at the end of summer (15.1+/-7.1 ng/ml in March; 21.5+/-5.5 ng/ml in June; 31.6+/-5.6 ng/ml in September; 23.1+/-5.3 ng/ml in December; mean+/-SD). The prevalence of hypovitaminosis D (<20 ng/ml) was 86.7%, 33.4%, 1.0%, and 26.0% in March, June, September, and December, respectively. Mean plasma intact parathyroid hormone (iPTH) concentration was lowest at the end of summer and highest at the end of winter (28.2+/-9.3 pg/ml in March; 21.7+/-7.0 pg/ml in June; 19.8+/-6.9 pg/ml in September; and 25.7+/-9.2 pg/ml in December; mean+/ SD). Serum 25-OHD was inversely associated with iPTH (coefficient, -0.223; r=0.251; P<0.001). Serum 25-OHD levels were higher in men than in women. The serum 25-OHD level was positively associated with age, body weight, and body mass index, but not with body fat content. These results suggest a high prevalence of hypovitaminosis D associated with elevation of iPTH in Japan, in winter, even in a sunny area. PMID- 15750692 TI - Dolomite supplementation improves bone metabolism through modulation of calcium regulating hormone secretion in ovariectomized rats. AB - Dolomite, a mineral composed of calcium magnesium carbonate (CaMg (CO3)2), is used as a food supplement that supplies calcium and magnesium. However, the effect of magnesium supplementation on bone metabolism in patients with osteoporosis is a matter of controversy. We examined the effects of daily supplementation with dolomite on calcium metabolism in ovariectomized (OVX) rats. Dolomite was administered daily to OVX rats for 9 weeks. The same amount of magnesium chloride as that supplied by the dolomite was given to OVX rats as a positive control. Histological examination revealed that ovariectomy decreased trabecular bone and increased adipose tissues in the femoral metaphysis. Dolomite or magnesium supplementation failed to improve these bone histological features. Calcium content in the femora was decreased in OVX rats. Neither calcium nor magnesium content in the femora in OVX rats was significantly increased by dolomite or magnesium administration. Urinary deoxypyridinoline excretion was significantly increased in OVX rats, and was not affected by the magnesium supplementation. Serum concentrations of magnesium were increased, and those of calcium were decreased, in OVX rats supplemented with dolomite or magnesium. However, there was a tendency toward decreased parathyroid hormone secretion and increased calcitonin secretion in OVX rats supplemented with dolomite or magnesium. Serum 1,25-dihydroxyvitamin D(3) and osteocalcin levels were significantly increased in the supplemented OVX rats. These results suggest that increased magnesium intake improves calcium metabolism in favor of increasing bone formation, through the modulation of calcium-regulating hormone secretion. PMID- 15750694 TI - Effect of age on body sway assessed by computerized posturography. AB - The swaying and postural instability frequently seen in elderly subjects had not been analyzed quantitatively in detail until the introduction of computerized posturography. In order to assess the changes of body sway with aging, we performed computerized posturography in 144 subjects (51 men and 93 women, between the ages of 22 and 88 years) without specific neurological or metabolic disorders. The total and timed track length of the center of gravity, reflecting the distance of sway, increased with advancing age, with a highly significant positive correlation, without marked sex differences. The total area covered by the track of the center of gravity (expressing the extent of sway) also showed a similar tendency. Track density per unit area, expressing the efficiency of postural control, in contrast, decreased with age, showing a significant negative correlation with age, but only when the subjects had their eyes open; this decrease did not occur when they had their eyes closed. The Romberg ratio, an index of exacerbation of sway on eye closure, showed little change with a tendency for slight alleviation of sway and improvement in the efficiency of its control. Computerized posturography appears to be a useful tool with which to analyze the mechanism of swaying associated with old age. PMID- 15750695 TI - Body mass index, calcium intake, and physical activity affect calcaneal ultrasound in healthy Greek males in an age-dependent and parameter-specific manner. AB - Quantitative ultrasound (QUS) is a peripheral bone densitometry technique that is rapidly gaining in popularity for the assessment of skeletal status. This study was carried out to examine the effect of anthropometric, dietary, physical activity, and other lifestyle factors on QUS parameters in healthy Greek males of various ages, including children (n=192), adults (n=106), and elderly (n=86) subjects. Calcaneal QUS measurements were performed with the Sahara device (Hologic), which measures broadband ultrasound attenuation (BUA) and speed of sound (SOS) through the os calcis. A composite parameter, the quantitative ultrasound index (QUI), and an estimate of heel bone mineral density (eBMD) were also derived. BUA correlated positively with height, weight, and body mass index, as well as waist and hip circumferences (P<0.001), but this was not the case for SOS. QUI and eBMD were inconsistently related with anthropometric characteristics. Overweight and obese males had significantly higher BUA than normal-weight subjects (P<0.05), but similar SOS, QUI, and eBMD; this held true for all age groups. Boys participating in organized physical activities had significantly higher SOS, QUI, and eBMD than those who did not (P<0.05), although BUA was similar in the two groups; no differences according to organized physical activity were detected in adults and the elderly. On the other hand, adult men devoting at least some time to non-organized physical activities had significantly higher QUS values than their non-exercising peers (P<0.05); no such effects, however, were seen in children and the elderly. Adult men with calcium intakes above 800 mg/day had significantly higher SOS, QUI, and eBMD than those consuming less calcium (P<0.05), and also tended towards higher BUA (P=0.079); no such differences were observed among children and elderly men. The effects of physical activity and calcium intake on heel QUS persisted even after controlling for body size. Overall, body weight was the sole significant positive determinant of BUA (beta=0.373; t=6.589; P<0.001), explaining approximately 14% of the total variance, while age was the sole significant negative determinant of SOS (beta= 0.198; t=-3.321; P=0.001), albeit explaining only less than 4% of the total variance. In conclusion, body size, dietary calcium intake, and physical activity patterns seem to inconsistently and age-dependently influence heel QUS among healthy Greek males in a parameter-specific manner. PMID- 15750696 TI - Health-related quality of life (HRQOL) in Japanese osteoporotic patients and its improvement by elcatonin treatment. AB - Health-related quality of life (HRQOL; "QOL" hereafter) was evaluated in Japanese osteoporotic patients using three questionnaires; the SF-36 (MOS 36-Item Short Form Health Survey; generic, profile-type), the EQ-5D (Euro Qol-5 Dimensions; generic, preference-based), and the JOQOL (Japanese Osteoporosis Quality of Life 1999; disease-targeted). The eight subscales and two summary scores of the SF-36 were impaired in these patients even after correction for age and sex. The scores on the EQ-5D and JOQOL correlated well with the subscales of the SF-36 that represent the physical aspects of physical function and bodily pain, which suggests that physical aspects are important determinants of overall QOL status in osteoporotic patients. Although the QOL scores did not correlate with bone mineral density, they were markedly influenced by the presence of vertebral fractures. In particular, the presence of two or more vertebral fractures greatly decreased the QOL scores. We then evaluated the QOL scores before and after treatment. The patients were either given calcium supplementation alone or calcium plus once-weekly elcatonin (Elcitonin, Asahi Kasei Pharma, Tokyo, Japan) injection. Elcatonin treatment markedly improved diverse aspects of the QOL, whereas calcium alone did not. The current data suggest that osteoporosis, especially in the presence of vertebral fracture, is associated with compromised QOL, and therapeutic intervention for osteoporosis should be evaluated in terms of QOL, as well as in terms of increases in bone mineral density and fracture prevention. PMID- 15750697 TI - Regional variations in microstructural properties of vertebral trabeculae with aging. AB - The aim of this study was to identify regional variations in the three dimensional microstructure of vertebral cancellous bones, and their relative differences with respect to aging. Ninety trabecular specimens were obtained from six normal L4 vertebral bodies of six male cadaver donors in two age groups, three aged 62 years and three aged 69 years; (n=45, each). In each vertebral body, five trabecular columns, each of 8x8x25 mm3, were cut from the anterior, posterior, central, right, and left regions. These columns were scanned, using high-resolution micro-computed tomography (microCT), three times, to obtain superior, middle, and inferior layers. Fifteen regions were obtained for each vertebral body. For all 90 trabecular specimens the bone volume fraction (BV/TV), trabecular number (Tb.N), and trabecular thickness (Tb.Th), as well as the three radii of the mean intercept length (MIL) ellipsoid (H1, H2, and H3) were determined. Regional variations in different transverse layers and vertical columns within and between the two age groups were then analyzed. The results showed significant differences in BV/TV, Tb.N, DA, and H2/H3 between the two age groups. The BV/TV and Tb.N were decreased, while the anisotropic parameters were increased significantly with age, increasing from age 62 to 69. Change in Tb.Th was not statistically significant, although the average was slightly smaller in the 69-year group. Each microstructural parameter followed its own pattern of regional variation within each group, suggesting both mechanical and age-related adaptation. This is the first study that has provided microstructural data of the vertebral body in a Chinese sample. These data may help us to gain more insight into the mechanism of the occurrence of lumbar osteoporosis and the related regional fracture risks, and may provide a reference for better enhancement of fracture repair. PMID- 15750698 TI - Genetic determination of variation and covariation of bone mineral density at the hip and spine in a Chinese population. AB - Bone mineral density (BMD) is a significant determinant of risk for osteoporosis. Genetic factors are known to account for a major proportion of variation of BMD in Caucasians. However, the degree of genetic determination of BMD in Chinese populations has seldom been investigated. The aim of our study was to investigate the magnitude of the genetic determination of BMD at the spine and hip, and their genetic covariation, in a population of Shanghai city in P. R. China. The subjects consisted of 44 full-sib pairs of females aged 19-43 years, 186 mother daughter pairs, and 270 nuclear families. For BMD at the spine and hip, the values for narrow-sense heritability h2 (+/-SE) were 0.72+/-0.14 and 0.87+/-0.14, respectively, when estimated by full-sib pairs, and 0.44+/-0.07 and 0.77+/-0.07, respectively, when estimated by mother-daughter pairs. There was a significant genetic correlation r(g) (+/-SE) of BMD between the spine and hip, of 0.97+/-0.01 and 0.76+/-0.04, respectively, when estimated by full-sib pairs and mother daughter pairs. The common household impact on BMD in our study was negligible according to the statistical estimate. We conclude that genetic factors play a major role in the determination of the variation and covariation of BMD at the spine and hip in our Chinese sample. PMID- 15750699 TI - Beneficial effects of regular Tai Chi exercise on musculoskeletal system. AB - This study was performed to evaluate the potential benefits of regular Tai Chi Chun (TCC) exercise on bone mineral density (BMD) and neuromuscular function in postmenopausal women. In this cross-sectional study, 99 healthy postmenopausal women, with a mean age of 55.9+/-3.1 years and within 10 years after the menopause, were recruited; including 48 subjects who had been regularly practicing TCC exercise for more than 3 h/week and 51 age- and sex-matched sedentary controls (CON). BMD was measured in the lumbar spine and proximal femur of the non-dominant leg (femoral neck, greater trochanter, and Ward's triangle), using dual-energy X-ray absorptiometry (DXA). Neuromuscular function was evaluated, including magnitude of trunk bend-and-reach, quadriceps muscle strength, and single-stance time on the nondominant leg. The TCC group showed overall higher BMD at all measurement sites, with a significant difference found at the spine (7.1%), greater trochanter (7.2%), and Ward's triangle (7.1%) of the proximal femur (all; P<0.05). Functional tests revealed an average 43.3% significantly greater quadriceps strength (P<0.01), and 67.8% significantly longer single-stance time in the TCC group as compared with the CON group (P<0.05), as well as a greater magnitude of trunk bend-and-reach in the TCC group (P=0.08). Bivariate linear correlation analysis showed that quadriceps muscle strength was significantly correlated with the single-stance time (r=0.41; P<0.01). This study revealed that regular TCC exercise may have an association with higher BMD and better neuromuscular function in early postmenopausal women. PMID- 15750700 TI - Relief of migraine following refractive surgery for anisometropia. PMID- 15750701 TI - Functional motor compensation in amyotrophic lateral sclerosis. AB - The present study investigated the fMRI correlates of functional compensation/neural reorganization of the motor system in patients with amyotrophic lateral sclerosis (ALS). The hypothesis was that ALS patients would recruit additional brain regions compared with controls in a motor task and that activity in these regions would vary as a function of task difficulty. Patients and controls executed a motor task with two sequences (a simple and a more difficult one) of consecutive button presses. Patients and controls both activated brain regions known to be involved in motor execution and control. Activity in ipsilateral motor areas as well as difficulty-related activity in the left cerebellum could only be observed in patients. The behavioral data indicated that the motor task was much more difficult for patients than for controls. At nearly equal difficulty the observed patterns of hemodynamic activity in controls were very similar to those observed in ALS. The findings suggest that functional compensation in ALS relies on existing resources and mechanisms that are not primarily developed as a consequence of the lesion. PMID- 15750702 TI - CSF pleocytosis and expansion of spinal lesions in Japanese multiple sclerosis with special reference to the new diagnostic criteria. AB - New diagnostic criteria for multiple sclerosis (MS) were recently proposed from the international panel on the diagnosis of MS, and they include exclusion criteria, such as lesions extending over more than two vertebral segments on spinal MRI and CSF pleocytosis of more than 50/mm3. We reviewed the clinical features of 158 patients who satisfied the diagnostic criteria for MS except for having the above atypical paraclinical findings. All patients exhibited two or more clinical attacks and objective clinical evidence of multiple lesions without any evidence of other disorders. Thirty-three (20.9%) patients had one or both atypical paraclinical findings. Twenty-one out of the 33 patients were classified as having optico-spinal MS (OSMS), and the other 12 as non-OSMS patients with atypical large expanding or destructive cerebral, cerebellar or brainstem lesions on MRI as well as one or both atypical paraclinical findings. Based on this heterogeneity in clinical findings in MS, there is an urgent need to develop a common general concept of the "MS" syndromes, and the ethnic-related heterogeneity should be considered in the revised criteria for the diagnosis of MS. PMID- 15750703 TI - CXCL11 is involved in leucocyte recruitment to the central nervous system in neuroborreliosis. AB - We investigated the role of the CXCR3 chemokine CXCL11 (I-TAC) for leukocyte recruitment to the CSF in neuroborreliosis (NB). CXCL11 levels in the CSF of 17 patients with acute NB were elevated compared with 20 non-inflammatory controls (100.1 vs. 54.1 pg/ml, p = 0.002). Using a modified Boyden chamber chemotaxis assay, the CSF of patients with acute NB was more chemotactic than the control CSF-samples (p = 0.001). A strong correlation (p < 0.001) between CXCL11 levels, chemotactic activity and CSF-white cell count was detected. Though the chemotactic activity of CSF from NB patients was only partially reduced by a human recombinant CXCL11 antibody, these findings suggest that, among other chemotactic factors, CXCL11 may play a role in the chemotaxis of mononuclear cells in NB. PMID- 15750704 TI - The impact of inpatient neurorehabilitation on psychological well-being on discharge and at 3 month follow-up. AB - INTRODUCTION: Neurological rehabilitation aims to improve quality of life of patients with acute and chronic neurological conditions. Much of the existing research focuses on the impact of rehabilitation on physical functioning, with less emphasis on emotional wellbeing. This study assessed changes in psychological functioning in patients on discharge from rehabilitation and three months after discharge. METHODS: Patients admitted over a six-month period to a neurological rehabilitation unit were recruited prospectively to this study. Psychological functioning was measured by the self-report General Health Questionnaire and the Hospital Anxiety and Depression Scale. In addition, physical functioning was measured by the Barthel index and Functional Independence Measure. RESULTS: Psychological functioning was found to be significantly improved with rehabilitation. However, after three months, patients' scores returned to pre-treatment values. Anxiety was consistently elevated on admission, discharge and follow-up. In contrast, physical functioning improved from admission to discharge and was maintained at follow-up assessment. CONCLUSION: Rehabilitation services need to focus more on psychological functioning after discharge and identify effective strategies to maintain wellbeing. PMID- 15750705 TI - Discriminant analysis of the cognitive performance profile of MS patients differentiates their clinical course. AB - OBJECTIVE: To compare the neuropsychological deficits of primary progressive multiple sclerosis with those of relapsing-remitting and secondary progressive multiple sclerosis. METHODS: Sixty-five patients with different clinical courses of MS were neuropsychologically tested for language, attention, memory and executive functions. Discriminant analysis was used to predict the type of clinical course either by clinical variables (age, EDSS and duration of illness) or neuropsychological test results. RESULTS: For single neuropsychological tests, group differences were rare between the progressive courses and the relapsing remitting course of MS or absent between the progressive courses of MS. However, discriminant analysis correctly identified 87.7 percent of the patients' courses in general, and about 90 percent of the patients with chronic progressive MS. CONCLUSION: Using discriminant analysis, this study found neuropsychological impairment characteristic for relapsing remitting, secondary progressive and primary progressive patients. PMID- 15750706 TI - Cerebrospinal fluid HIV viral load in different phases of HIV-associated brain disease. AB - We compared CSF HIV viral load in 33 asymptomatic HIV seropositive patients, 11 patients with incipient minor motor deficits (MMD), 11 patients with sustained MMD, and 16 patients with HIV-associated dementia. Patients with incipient MMD showed significantly higher CSF viral load than asymptomatic patients. Demented patients also had higher CSF viral loads than asymptomatic patients. This phenomenon is independent of antiretroviral therapy. Thus, correlation of viral load with time suggests a multiphasic course of HIV-associated CNS disease. PMID- 15750707 TI - Activation of secondary auditory cortex in a deaf patient during song hallucinosis. PMID- 15750708 TI - Corticomotor organisation and motor function in multiple sclerosis. AB - Our objective was to determine whether there are changes in the corticomotor map for the hand in multiple sclerosis, and whether these changes correlate with indices of motor function and measures of corticomotor conduction or excitability. Transcranial magnetic stimulation (TMS) maps, motor evoked potential (MEP) latency and amplitude, motor threshold and EDSS and Purdue pegboard measurements were made in 26 subjects with relapsing-remitting multiple sclerosis. Correlations were sought between these measurements using the Pearson product-moment correlation with a level of significance of p = 0.05 (two-tailed). Map displacement was positively correlated with MEP latency (p = 3 x 10(-4)) and EDSS (p = 0.007), and negatively correlated with Purdue score (p = 4 x 10(-4)). Purdue scores correlated with all MEP parameters (latency, p = 4 x 10(-10); threshold, p = 4 x 10(-6); amplitude, p = 0.003). We conclude that motor reorganisation is associated with impaired corticomotor conduction and may reflect a process of neural plasticity associated with axonal demyelination in MS. An understanding of motor function in MS should incorporate models of both axonal demyelination and conduction deficits as well as neural plasticity. PMID- 15750709 TI - Spontaneous thoracic extradural haematoma presenting as the Brown-Sequard syndrome. PMID- 15750710 TI - Paroxysmal raised intracranial pressure associated with spinal meningeal cysts. AB - Raised intracranial pressure in association with spinal meningeal cysts has rarely been reported. We describe four patients in whom evidence of paroxysmal raised intracranial pressure was found in association with spinal meningeal cysts. Cerebrospinal fluid diversion procedures have previously been shown to relieve local symptoms due to spinal cysts. In our patients symptoms of paroxysmal headache were alleviated by this method, suggesting a causal relationship with the raised pressure. This association may be an under diagnosed cause of paroxysmal headaches. We review the medical literature on the classification of spinal meningeal cysts, evaluate the theories of their origin and offer suggestions on the pathogenesis of the abnormal CSF dynamics that may allow an interplay between raised intracranial pressure and spinal meningeal cysts to produce paroxysmal symptoms. PMID- 15750711 TI - Sputum eosinophilia in idiopathic pulmonary fibrosis. AB - OBJECTIVES AND DESIGN: Cough is a common symptom in idiopathic pulmonary fibrosis that is difficult to treat and has a major impact on quality of life. We tested the hypothesis that the cough and increased cough reflex sensitivity seen in patients with idiopathic pulmonary fibrosis may be due to airway inflammation in a prospective, cross-sectional study. SUBJECTS AND METHODS: We measured the induced sputum inflammatory cell profile and cell-free supernatant inflammatory mediator concentrations in 15 patients with idiopathic pulmonary fibrosis, 17 healthy controls and 15 patients with chronic obstructive pulmonary disease. RESULTS: Both the geometric mean sputum differential eosinophil cell count and median eosinophilic-cationic-protein concentration were significantly higher in patients with idiopathic pulmonary fibrosis than controls (2.1% vs 0.3%; p <0.001 and 1.1 mg/ml versus 0.2 mg/ml; p=0.03 respectively). There were no significant differences in sputum eosinophil counts and eosinophilic-cationic-protein concentrations between patients with idiopathic pulmonary fibrosis and chronic obstructive pulmonary disease. Sputum leukotriene-B4 concentrations were significantly lower in patients with idiopathic pulmonary fibrosis (p=0.03) and chronic obstructive pulmonary disease (p=0.008) compared to controls. CONCLUSIONS: Idiopathic pulmonary fibrosis is characterised by the presence of active eosinophilic airway inflammation raising the possibility that airway inflammation may contribute to symptoms such as cough. PMID- 15750712 TI - Reversible and irreversible airway inflammation and fibrosis in mice exposed to inhaled ovalbumin. AB - OBJECTIVE AND DESIGN: We examined the reversibility of several changes in the lungs and airways of mice immediately after exposure to ovalbumin aerosol and after a period of recovery breathing clean air. METHODS: Mice were exposed for 1, 2, 4, 6, 8, or 10 weeks, with recovery in clean air for 1-3 weeks. RESULTS: Airway collagen content, exhaled NO, airway mucous cell hyperplasia, and lung lavage inflammatory cell content increased upon exposure to ovalbumin aerosol. All parameters except airway fibrosis decreased partially or completely to control values with recovery in clean air. CONCLUSIONS: Airway mucous cell hypertrophy and hyperplasia appear to be completely reversible after recovery in clean air, while exhaled NO and airway inflammation appear to be mostly reversible, except for persistence of lymphocytes in the lung lavage fluid. Airway fibrosis appears to be reversible when mice are exposed to ovalbumin aerosol for periods of up to 4 weeks of exposure, but becomes irreversible after 6 or more weeks of exposure. PMID- 15750713 TI - Augmentation of the bactericidal activities of human cathelicidin CAP18/LL-37 derived antimicrobial peptides by amino acid substitutions. AB - OBJECTIVE: Mammalian myeloid and epithelial cells express various peptide antibiotics (such as defensins and cathelicidins) that contribute to the innate host defense against invading micro-organisms. Among these, human cathelicidin CAP18/LL-37 (L1-S37) possesses potent antibacterial activities against Gram positive and Gram-negative bacteria. In this study, to develop peptide derivatives with improved bactericidal actions, we utilized the amphipathic 18 mer peptide (K15-V32) of LL-37 as a template, and evaluated the activities of modified peptides. METHODS: Antibacterial activities of the peptides (0.022 approximately 4.4 microM corresponding to 0.1 approximately 10 microg/ml) were assessed by alamarBlue assay using Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Escherichia coli and Pseudomonas aeruginosa as target organisms. Furthermore, the membrane-permeabilization activities of the peptides were examined by using E. coli ML-35p as a target. RESULTS: By substituting E16 and K25 with two L residues, the hydrophobicity of the peptide (18-mer LL) was increased, and by further substituting Q22, D26 and N30 with three K residues, the cationicity of the peptide (18-mer LLKKK) was enhanced. Among peptide derivatives, 18-mer LLKKK exhibited the most potent antibacterial actions against S. aureus (methicillin-resistant and -sensitive), S. pneumoniae, S. pyogenes, E. coli and P. aeruginosa, and possessed the most powerful membrane permeabilizing activities against E. coli ML-35p at the effective concentrations (p <0.05, 18-mer LLKKK vs. 18-mer LL, 18-mer K15-V32 and LL-37). CONCLUSIONS: Bactericidal activities of the amphipathic human CAP18/LL-37-derived 18-mer peptide can be augmented by modifying its hydrophobicity and cationicity, and 18 mer LLKKK is the most potent among peptide derivatives with therapeutic potential for Gram-positive and Gram-negative bacterial infections. PMID- 15750714 TI - Dialyzable leukocyte extract differentially regulates the production of TNFalpha, IL-6, and IL-8 in bacterial component-activated leukocytes and endothelial cells. AB - OBJECTIVE: To investigate i) whether the Dialyzable Leukocyte Extract (DLE) modulates the production of proinflammatory cytokines in leukocytes activated by the bacterial cell wall components lipopolysaccharide (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN); ii) the effect of DLE on LPS-stimulated endothelial cells; and iii) whether the regulatory effect of DLE on inflammatory mediators is related to the modulation of Toll-like receptors (TLRs), NF-kappaB and cAMP signaling pathways. METHODS: Leukocytes were stimulated with LPS, LTA, and PGN in the presence of DLE. Endothelial cells were stimulated with LPS and treated with DLE. The levels of Tumor Necrosis Factor-alpha(TNFalpha), Interleukin-6 (IL-6), and IL-8 in culture supernatants were evaluated by ELISA. The expression of Toll-like receptor 2 (TLR2) and 4 (TLR4), NF-kappaB activity and cAMP levels were evaluated by flow cytometry, EMSA, and EIA, respectively. RESULTS: The addition of DLE to leukocytes stimulated with cell wall constituents suppressed the production of TNFalpha. However, DLE induced IL-8 release in monocytes and enhanced IL-6 and IL-8 production by activated monocytes and endothelial cells. Also, DLE induced TLR2 and TLR4 expression, and increased cAMP levels, whereas NF-kappaB activity was inhibited. CONCLUSIONS: The present data indicate the differential regulation by DLE of the production of TNFalpha, IL-6, and IL-8 cytokines, associated with effects on TLR2 and TLR4 expression and NF kappaB and cAMP activities. We suggest a putative mechanism for the biological effects of DLE in activated leukocytes and endothelial cells. PMID- 15750715 TI - Agonist and antagonist dual effect of the cross-linked S19 ribosomal protein dimer in the C5a receptor-mediated respiratory burst reaction of phagocytic leukocytes. AB - OBJECTIVE: To examine the behavior of the crosslinked dimer of S19 ribosomal protein (RP S19), a natural C5a receptor ligand, in the C5a receptor-mediated respiratory burst reaction of monocytes and neutrophils. METHODS: The respiratory burst reaction of leukocytes was quantitatively observed by continuous spectrophotometric measurement of the reduction of a water-soluble tetrazolium salt, WST-1. RESULTS: The RP S19 dimer induced the respiratory burst of monocytes, but not of neutrophils. Furthermore, in neutrophils, the RP S19 dimer inhibited the reaction induced by C5a, but did not affect the formyl-Met-Leu-Phe induced reaction. The dimer of a deletion mutant at the C-terminal portion of the RP S19 induced a respiratory burst reaction similar to the one induced by C5a, both in monocytes and neutrophils. Inversely, a chimeric fusion protein between C5a and RPS19, consisting of the addition of the 12 C-terminal amino acid residues of RP S19 after the C-terminal Arg74 of the C5a molecule, behaved similarly to the RP S19 dimer. CONCLUSIONS: The RP S19 dimer works as an agonist and antagonist of the C5a receptor in the monocyte and the neutrophil respiratory burst reactions, respectively. The switch moiety between the antagonist and agonist of the RP S19 dimer for the C5a receptor in the induction of respiratory burst of phagocytes localizes at the C-terminal region of RP S19. PMID- 15750716 TI - Intracellular production of IL-2, IL-4 and IFN-gamma by peripheral blood CD3+ cells in intermittent allergic rhinitis. AB - BACKGROUND: Allergic inflammation is mainly driven by type 2 T helper cells. The aim was to assess the changes in production of type 1 and 2 cytokines by CD3+ T cells dependent on natural exposure to allergens in subjects with intermittent allergic rhinitis (IAR) and in non-atopic subjects. MATERIAL: A total of 13 patients with IAR and 13 healthy non-atopics were recruited into the study. 11 patients with IAR were examined during the grass pollen season and 11 patients outside the season, 9 of them were assessed on both occasions. METHODS: A flow cytometric assessment of intracellular expression of IL-2, IL-4 and IFN-gamma by CD3+ cells was performed. For statistical analysis non-parametric tests were used. RESULTS: A tendency to decreased production of IL-4 outside the season was observed (6.94% [3.42-13.33] in season vs. 2.06% [0.7-3.6] out of season). The production of IL-4 was higher in the rhinitic group in the season than in the control group (1.93% [1.07-4.97], p=0.0034) and production of IL-2 was higher both in and outside the season (9.1% [3.94-15.09] and 10.0% [4.79-25.35] vs. 3.64% (2.64-5.03), p=0.037 and 0.045, respectively). IL-4/IL-2 and IL-4/IFN-gamma ratios were higher in the IAR group in the season than outside the season. CONCLUSION: A tendency towards a switch from a predominant type 2 response during natural allergen exposure to its suppression outside the season was found, together with a stable type 1 response. PMID- 15750717 TI - A possible etiology of the internal derangement of the temporomandibular joint based on the MRI observations of the lateral pterygoid muscle. AB - The purpose of this study was to evaluate the relationship of the temporomandibular joint (TMJ) internal derangement and lateral pterygoid muscle (LPM) by magnetic resonance imaging (MRI). In this study, 115 subjects with TMJ internal derangement (total 230 TMJs) and 21 subjects without clinical symptoms (total 42 TMJs) were included. TMJ disc position and LPM were evaluated using MRI. LPM attachments were categorized into two different types: type 1, where fibers of the superior head of the LPM (SLPM) were attached to the disc and fibers of the inferior head of the LPM (ILPM) were attached to condyle, and type 2, where fibers of the SLPM were attached to the disc and condyle, and fibers of the ILPM were attached to condyle. The presence of muscle atrophy and degeneration were also evaluated. LPM attachments were observed in two different parts. Disc displacements were common in the muscle attachments of both types. Type 1 muscle attachments were seen in 85.9% of all the anterior disc displacement without reduction (ADD) TMJs (total 64 TMJs). Atrophy was seen in a higher proportion (43.7%) in TMJs with ADD (28/64) than in TMJs with normal and anterior disc displacement with reduction (ADDR). Out of 74 TMJs with atrophy, 68 had type 1 muscle attachment. Four TMJs had atrophy in both superior and inferior heads of the lateral pterygoid. However, atrophy was not present only in the ILPM. It has been concluded that since the SLPM only attached to the disc in type 1, the disc may displace anteriorly very easily. Therefore, this situation will reduce the function of the SLPM. Reduced muscle function may cause muscle atrophy. The activity of the SLPM may be more reduced since the disc permanently dislocated in TMJs with ADD. Finally, spasm of the LPM causes disc displacement and atrophy and then the degeneration of the LPM may follow disc displacement. PMID- 15750721 TI - Results from high-throughput DNA cloning of Arabidopsis thaliana target genes using site-specific recombination. AB - The Center for Eukaryotic Structural Genomics (CESG) was founded as a collaborative effort to develop technologies for the rapid and economic determination of protein three-dimensional structures. The initial focus was on the genome of the model plant Arabidopsis thaliana. Protocols for high-throughput cloning of Arabidopsis open reading frames into Escherichia coli expression vectors are presented along with an analysis of results from approximately 2000 cloning experiments. Open reading frames were chosen on the likelihood that they would represent important unknown regions of protein conformation and fold space or that they would elucidate novel fold-function relationships. The chosen open reading frames were amplified from a cDNA pool created by reverse transcription of RNA isolated from an Arabidopsis callus culture. A novel Gateway protocol was developed to insert the amplified open reading frames into an entry vector for storage and sequence determination. Sequence verified entry clones were then used to create expression vectors again via the Gateway system. PMID- 15750726 TI - Mitochondrial functioning of constitutive iron uptake mutations in Cryptococcus neoformans. AB - Randomly obtained, constitutive plasma membrane ferric reductase/ferrous uptake mutants of Cryptococcus neoformans were mapped to four distinct loci by meiotic analysis. One of those loci, FRR1 , was previously found homologous to MRS3 and MRS4 of Saccharomyces cerevisiae , which determine proteins involved in mitochondrial transport of iron. We were able to complement, clone, sequence and thereby identify two of the three remaining constitutive uptake loci. FRR3 was found to be homologous to ISU1 and ISU2 of S. cerevisiae, which form mitochondrial iron-sulfur complexes; FRR4 was found to be homologous to YFH1, the yeast frataxin homologue, which also participates in iron-sulfur cluster biogenesis. Because of the constitutive iron uptake seen in these mutants, mitochondria appear to have a central role in the cellular iron economy; moreover, as judged by our mutational statistics, the genetic machinery for mitochondrial iron accumulation may be more complex than that of the cytoplasm. PMID- 15750727 TI - Human cryptococcosis: relationship of environmental and clinical strains of Cryptococcus neoformans var. neoformans from urban and rural areas. AB - Forty-five clinical and 55 environmental strains of Cryptococcus neoformans var. neoformans from Sao Paulo, Brazil, were tested for their susceptibilities to amphotericin B, fluconazole, itraconazole, and flucytosine by the broth microdilution method according to the National Committee of Clinical Laboratory Standards guidelines. Electrophoretic karyotypes analysis by counter-clamped homogeneous electrophoresis was used to compare their genetic relatedness. Molecular typing revealed three clinical profiles very similar to two environmental profiles and an identical environmental and clinical profile. The results showed that human cryptococcosis can be acquired from environmental strains, which had similar minimum inhibitory concentration values to clinical strains, for antifungal agents. PMID- 15750728 TI - Keratinophilic fungi isolated from hospital dust and soils of public places at Gulbarga, India. AB - Keratinophilic fungi were isolated from corridor dust of 11 hospitals and soils of 21 public places using a hair baiting technique. A total of 41 species belonging to 24 genera were recovered. Among the dermatophytes and related species, Microsporum gypseum was predominant, followed by Chrysosporium keratinophilum, Trichophyton mentagrophytes, M. nanum and Ch. tropicum. Other species were represented by 32 species belonging to 21 genera. Most of the species isolated are known to be agents of human and animal infection or have been isolated from human and animal lesions. The fungi observed here are discussed in relation to their global distribution. PMID- 15750729 TI - The susceptibility of immature stages of Bemisia tabaci to the entomopathogenic fungus Lecanicillium muscarium on tomato and verbena foliage. AB - Lecanicillium muscarium is a widely occurring entomopathogenic fungus. Laboratory studies were conducted to determine the efficacy of L. muscarium against different instars of Bemisia tabaci on tomato and verbena foliage after two incubation times (3 and 7 days). Significant reduction in B. tabaci numbers were recorded on fungus treated plants (p < 0.001). Second instar B. tabaci proved most susceptible to L. muscarium infection. There was no significant difference in mortality of B. tabaci second instars after either 3 or 7 days exposure to L. muscarium on either host plant. The importance of the speed of pest mortality following treatment and the potential of L. muscarium to be incorporated into an integrated pest management strategy for the biocontrol of B. tabaci on tomato and verbena plants are discussed. PMID- 15750730 TI - Fumonisin production by Fusarium species isolated from freshly harvested corn in Iran. AB - Fifty-one strains of Fusarium verticillioides and F. proliferatum isolated from corn collected from four different geographic areas in Iran, namely Fars, Khuzestan, Kermanshah and Mazandaran (an endemic oesophageal cancer (OC) area) were evaluated for their ability to produce fumonisins B1 (FB1), B2 (FB2) and B3 (FB3) in corn culture. Fumonisin levels were determined by high-performance liquid chromatography. All tested strains of F. verticillioides and F. proliferatumproduced fumonisins within a wide range of concentrations, 197-9661 microg/g, 18-1974 microg/g, and 21-1725 microg/g for FB1, FB2, and FB3, respectively. The highest mean concentrations of FB1, FB2, and FB3 were 3897, 806 and 827 microg/g, respectively. Overall, 61% of the F. verticillioides and F. proliferatum strains produced higher levels of FB3 than FB2. The mean ratios of FB1:FB2, FB1:FB3 and FB1:total fumonisins were 8, 7 and 0.7 for F. verticillioides and 5.7, 10.7 and 0.7 for F. proliferatum, respectively. Significant differences in some of the meteorological data (rainfall, relative humidity and minimum temperature) from the four provinces were observed. Fumonisin levels produced by F. verticillioides strains isolated from Khuzestan province (tropical zone) were significantly (P < 0.01) higher than the other three provinces. This is the first report of the fumonisin-producing ability of F.verticillioides and F. proliferatum strains isolated from corn harvested from different geographic areas in Iran. PMID- 15750731 TI - Microbiological and physicochemical factors affecting Aspergillus section Flavi incidence in Cavendish banana (Musa cavendishii) chips production in Southern Philippines. AB - Microbiological and physicochemical factors affecting the incidence of Aspergillus section Flavi in dried Cavendish banana (Musa cavendishii) chips production in Southern Philippines were examined. The average counts of Aspergillus section Flavi (AFC) in fresh and dried Cavendish bananas from 10 production batches of the Philippine Agro-Industrial Development Cooperative in Davao del Norte, Southern Philippines were 1.2 x 10(2) and 1.6 x 10(2) cfu/g, respectively. Isolates from both samples were identified to be Aspergillus flavus based on spore type and conidial structure of isolates. An increasing trend in the AFC of Cavendish bananas was observed during dried banana chips processing. Variability in the AFC between production batches was attributed to differences in aerobic and fungal populations and physicochemical characteristics of the fruits, peel damage of the raw materials, concentration of AFC in the air and food-contact surfaces of the production area, and temperature and relative humidity (RH) conditions of the environment during production and storage. Physicochemical characteristics of Cavendish bananas from the receipt of raw materials up to the first day of drying were within the reported range of values allowing growth and toxin production by aflatoxigenic fungi. Air-borne AFC varied depending on the section of the production area examined. The close proximity of the waste disposal area from the production operation to the preparation, drying and storage areas suggests that cross-contamination, probably air-borne or insect borne was a likely occurrence. The hands of workers were also identified as AFC sources. Results of this study highlight the need for the development of strategies to control aflatoxigenic fungi and aflatoxin contamination in Philippine dried Cavendish bananas. PMID- 15750732 TI - Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes) as a potential mycoparasite on Sphaerotheca fuliginea (Ascomycotina: Erysiphales). AB - Hyphomycete Paecilomyces fumosoroseus that is well known as saprophytic and entomopatogenic fungus was investigated for its mycoparasitism on the cucumber powdery mildew pathogen. Mycoparasitism was documented by using standard bioassay and SEM. Effects of mycoparasitism were evaluated in three types of experiments. Paecilomyces fumosoroseus was applied in the form of graded suspensions into a colony of powdery mildew on a leaf segment. Interaction between both fungi was observed as the percentage of colonized area vs. experimental time. In the second experiment, young cucumber plants were sprayed with a suspension of Paecilomyces fumosoroseus 24 h before inoculation of Sphaerotheca fuliginea. Pre-treatment with P. fumosoroseus reduced development and spreading of powdery mildew infection significantly 15 days post-inoculation in contrast to pre-treatments with sulfur fungicide and distilled water. The development of pure culture powdery mildew under determined experimental conditions was observed and compared with treated variants. In the third experiment, mildewed plants were treated with a suspension of P. fumosoroseus. The control treatments with sulfur fungicide and distilled water were tested. Effects of P. fumosoroseus on the dispersion of powdery mildew during a 21-day period were observed. P. fumosoroseus suppressed the development and spread of cucumber powdery mildew significantly during the time of the experiment. The mechanical and physical damages and disruptions of vegetative and fruiting structures of powdery mildew were recorded under light microscopy and S.E.M. Results were concluded in pursuance to differences between the natural behaviour and development of S. fuliginea on cucumber plants treated with P. fumosoroseus and non-treated plants. PMID- 15750733 TI - Field performance of maize grown from Fusarium verticillioides-inoculated seed. AB - Fusarium verticillioides is an important fungus occupying dual roles in the maize plant. The fungus functions as an endophyte, a fungal/host interaction beneficial to the growth of some plants. At other times, the fungus may function as a mycotoxin producing pathogen. The advantages and/or disadvantages of the endophytic relationship must be established in order to target appropriate sites for controlling diseases and mycotoxins in maize. One possibility could be to ensure seed maize is fungal free prior to planting. Reciprocal inoculations were made with two fungal isolates on seed of two maize genotypes. Yield was measured at harvest by ear and seed characters and vegetative growth at one-month intervals for plant survival, height, weight and stem diameter. Yield and vegetative growth differed among mature plants only once based on seed inoculation status. In 1998, plant weight was reduced and seed weight per ear was increased for the dent maize, GT-MAS: gk, grown from F. verticillioides RRC 374 inoculated seed compared to other seed treatments. Most vegetative characters were reduced at the first collection for Silver Queen plants grown from F. verticillioides-inoculated seed in 1997 and 1999, but not in 1998. However, no significant differences occurred among mature Silver Queen plants during any of the three growing seasons. In conclusion, yield and vegetative growth of mature maize plants grown from F. verticillioides-inoculated seed were equal to or greater than plants grown from non-inoculated seed under south Georgia field conditions during 1997, 1998, and 1999. PMID- 15750734 TI - First report of Phoma sorghina (Sacc.) Boerema Dorenbosch & van Kest on wheat leaves (Triticum aestivum L.) in Argentina. AB - A new disease caused by Phoma sorghina has been detected for the first time on wheat plants in the Province of Buenos Aires, Argentina. The pathogen was isolated from wheat leaves growing under field conditions, cultured on PDA and identified by its morphobiometric and cultural characters. The disease symptoms and morphological characters of the pathogen are described. Pathogenicity of the isolate was confirmed by inoculating 10 wheat cultivars under greenhouse conditions. PMID- 15750738 TI - Proceedings of the Third International Workshop on Ganoderma Diseases of Perennial Crops. March 24-26, 2003. Indonesia. PMID- 15750735 TI - Identification and pathogenic characterization of endophytic Fusarium species from cowpea seeds. AB - Isolates of Fusarium were obtained and identified from seeds of cowpea, Vigna unguiculata (L.) Walp., by means of blotter tests and slide cultures. Species were differentiated according to the morphology of the macroconidia, microconidia and their arrangement in chains or false heads, the size and type of conidiophore, and the presence or absence of chlamydospores. The species were identified as F. semitectum, F. equiseti, F. oxysporum, F. solani, F. anthophilum, F. sporotrichioides, F. moniliforme, and Fusarium sp. Among the species, F. semitectum was the most frequently detected. None of these species were pathogenic when inoculated in susceptible cowpea cultivar (BR 17-Gurgueia). But, an isolate of F. oxysporum f. sp. tracheiphilum used as a standard of comparison for pathogenicity (control) induced symptoms of yellowing, vascular wilting, and death of a susceptible cowpea cultivar under the same environmental conditions. PMID- 15750739 TI - Possible sources of genetic resistance in oil palm (Elaeis guineensis Jacq.) to basal stem rot caused by Ganoderma boninense--prospects for future breeding. AB - Oil palm estates in southeast Asia suffer from substantial losses due to basal stem rot caused by Ganoderma boninense. Field observations have been carried out in North Sumatra, Indonesia, on a series of planting materials of known origin. Differences in susceptibility to the disease have been detected within the two Elaeis species, guineensis and oleifera. Within Elaeis guineensis, material of Deli origin is highly susceptible compared to material of African origin. It is also possible to detect differences in reaction between parents and between crosses within a given origin. The variability of resistance to basal stem rot within the same cross is also illustrated by the diverse responses of clones derived from palms of the same origin. The prospects opened up by these results are discussed, and the importance of performing an early selection test is highlighted. PMID- 15750740 TI - Studies on oil palm trunks as sources of infection in the field. AB - Diseases of oil palm caused by Ganoderma boninense are of major economic importance in much of South-East Asia. This paper describes results from an ongoing field trial concerning the spread of the pathogen from artificially inoculated trunks used to simulate spread from windrowed trunks. Three planting distances for bait seedlings revealed that the closer the seedling was planted to the source of inoculum the sooner it succumbed to the disease. However, infection only occurred when the trunks were mounded (covered with soil), and seedlings planted around uncovered trunks (at any distance) have showed no symptoms of disease to date. Isolates are being collected from infected plants and molecular analysis is being undertaken to give more information on the spread of the pathogen. PMID- 15750741 TI - Investigations on the causes of upper stem rot (USR) on standing mature oil palms. AB - Three different trials to examine the cause of upper stem rot (USR) infection in oil palm failed to achieve any infection. In the first experiment, inoculum was applied as colonised rubber wood blocks or as spore suspensions. In the second experiment, particular attention was given to ensure that the Ganoderma spores were freshly collected to maintain viability but no infection was observed around the inoculation sites of any of the different oil palm tissues treated. Lastly in the third experiment, both monokaryotic and dikaryotic mycelial cultures were applied directly to cut fronds, which were protected with a moist covering, but no infection was detected after more than two years. Failure to achieve infection by direct inoculation would indicate that USR does not arise from direct infection of living tissues by Ganoderma spores or mycelium, this is probably because of insufficient inoculum potential to cause infection. It is suggested that USR infection is achieved only when a sufficiently large source of inoculum has built up in dead material, probably in frond axils, and this allows invasion of the living tissues. PMID- 15750742 TI - Quantification and characterisation of Trichoderma spp. from different ecosystems. AB - Basal stem rot of oil palm caused by Ganoderma boninense is of major economic importance. Observations of the low incidence of disease due to Ganoderma species in natural stands, suggest that the disease is kept under control by some biological means. Trichoderma spp. are saprophytic fungi with high antagonistic activities against soil-borne pathogens. However, their abundance and distribution are soil and crop specific. Trichoderma species have been found to be concentrated in the A1 (0-30 cm) and Be soil horizons (30-60 cm), although the abundance of Trichoderma was not significantly different between the oil palm and non-oil palm ecosystems. Characterisation of Trichoderma isolates based on cultural, morphological and DNA polymorphism showed that T. harzianum, T. virens, T. koningii and T. longibrachiatum made up 72, 14, 10 and 4% of the total Trichoderma isolates isolated. As Trichoderma species are present in the oil palm ecosystem, but at lower numbers and in locations different from those desired, soil augmentation with antagonistic Trichoderma spp. can be developed as a strategy towards integrated management of basal stem rot of oil palm. PMID- 15750743 TI - Diseases caused by Ganoderma spp. on perennial crops in Pakistan. AB - Ganoderma applanatum (Pres. Wallr) Pat. and G. lucidum (Leyss. ex Fr.) Karst attack species of Pinus, Dalbergia, Artocarpus, Morus, Cedrus, Melia, Quercus, Populus and other trees in Pakistan causing stem, butt and root rot diseases. A research institution to manage the diseases of perennial crops in general and of trees yielding edible oil in particular such as coconut and oil palm needs to be established in Pakistan. PMID- 15750744 TI - Following basal stem rot in young oil palm plantings. AB - The PCR primer GanET has previously been shown to be suitable for the specific amplification of DNA from Ganoderma boninense. A DNA extraction and PCR method has been developed that allows for the amplification of the G. boninense DNA from environmental samples of oil palm tissue. The GanET primer reaction was used in conjunction with a palm-sampling programme to investigate the possible infection of young palms through cut frond base surfaces. Ganoderma DNA was detected in frond base material at a greater frequency than would be expected by comparison with current infection levels. Comparisons are made between the height of the frond base infected, the number of frond bases infected, and subsequent development of basal stem rot. The preliminary results suggest that the development of basal stem rot may be more likely to occur when young lower frond bases are infected. PMID- 15750745 TI - Stem rots of oil palm caused by Ganoderma boninense: pathogen biology and epidemiology. AB - Oil palm (Elaeis guineensis Jacq.) has been grown in Papua New Guinea since the early 1960s. The most important disease of oil palm in PNG is a stem rot of the palm base. This is the same disease that constitutes a major threat to sustainable oil palm production in SE Asia. Investigations into the causal pathogen have revealed that the stem rots in PNG are caused predominantly by the basidiomycete Ganoderma boninense, with a minor pathogen identified as G. tornatum G. tornatum was found to have a broad host range whereas G. boninense appears to be restricted to palms. The population structure of G. boninense was investigated using inter-fertility studies between isolates collected from basal stem rots on oil palm. Although the G. boninense field populations are predominantly comprised of distinct individuals, a number of isolates were found that share single mating alleles. This indicates that out-crossing had occurred over several generations in the resident or wild population of G. boninense prior to colonization of oil palm. No direct hereditary relationship between isolates on neighbouring diseased palms was found, although an indirect link between isolates causing upper stem rot and basal stem rot was detected. PMID- 15750746 TI - An insight into spore dispersal of Ganoderma boninense on oil palm. AB - The disease of oil palm caused by Ganoderma boninense, although universally referred to as Ganoderma basal stem rot, occurs in three very distinct phases, with basal stem rot only part of the disease cycle. G. boninense also causes a seedling disease and an upper stem rot. An understanding of spore dispersal provides an insight into where spores of G. boninense have a role in the infection process. This role will be discussed in relation to each of these three infection phases. This understanding is a critical component of developing a successful disease control strategy. PMID- 15750747 TI - Ganoderma diseases of perennial crops in India--an overview. AB - The species of Ganoderma recorded from India as causing diseases of perennial crops are listed, and their host range and taxonomy discussed. Four new hosts of G. lucidum are also reported. A decline in productivity and the death of trees are the main economic impacts due to Ganoderma diseases, and the fungus is identified as a serious pathogen of cash crops, forest plantations and trees in natural forests in the country. Ganoderma diseases have been recorded on 144 hosts in India, the major pathogens being G. lucidum and G. applanatum. G. lucidum has been recorded on 91 hosts, and appears to cause the most widespread diseases. Identification has largely been made from morphological and cultural characters, and the names currently in use should therefore be treated with caution. Cultural methods of disease control are largely inefficient in minimising inoculum pressure and in reducing the disease incidence. Chemical methods in combination with soil amendments form short-term solutions for managing the disease and improving productivity. The immediate priorities for developing an efficient management system for Ganoderma diseases in India are: (1) a thorough understanding of the etiology and epidemiology of the diseases on different hosts, (2) clarifying current ambiguity in species names, (3) assessing the inter-relationships between populations of Ganoderma on different hosts and (4) developing tools for early detection of diseases in important crops. PMID- 15750748 TI - Enhancing biological control of basal stem rot disease (Ganoderma boninense) in oil palm plantations. AB - Basal Stem Rot (BSR) disease caused by Ganoderma boninense is the most destructive disease in oil palm, especially in Indonesia and Malaysia. The available control measures for BSR disease such as cultural practices and mechanical and chemical treatment have not proved satisfactory due to the fact that Ganoderma has various resting stages such as melanised mycelium, basidiospores and pseudosclerotia. Alternative control measures to overcome the Ganoderma problem are focused on the use of biological control agents and planting resistant material. Present studies conducted at Indonesian Oil Palm Research Institute (IOPRI) are focused on enhancing the use of biological control agents for Ganoderma. These activities include screening biological agents from the oil palm rhizosphere in order to evaluate their effectiveness as biological agents in glasshouse and field trials, testing their antagonistic activities in large scale experiments and eradicating potential disease inoculum with biological agents. Several promising biological agents have been isolated, mainly Trichoderma harzianum, T. viride, Gliocladium viride, Pseudomonas fluorescens, and Bacillus sp. A glasshouse and field trial for Ganoderma control indicated that treatment with T. harzianum and G. viride was superior to Bacillus sp. A large scale trial showed that the disease incidence was lower in a field treated with biological agents than in untreated fields. In a short term programme, research activities at IOPRI are currently focusing on selecting fungi that can completely degrade plant material in order to eradicate inoculum. Digging holes around the palm bole and adding empty fruit bunches have been investigated as ways to stimulate biological agents. PMID- 15750749 TI - Identification of Ganoderma, the causal agent of basal stem rot disease in oil palm using a molecular method. AB - From comparison of the alignments of the internally transcribed spacers (ITS) of ribosomal DNA from Ganoderma associated with oil palm basal stem rot (BSR) and other Ganoderma species, two specific primer pairs were selected to provide a specific DNA amplification of pathogenic Ganoderma in oil palm. Each primer pair produced a single PCR product of about 450 bp (for primer pair IT1-IT2) and 334 bp (for primer pair IT1-IT3) when oil palm Ganoderma DNA was used. No PCR amplification product was observed when other Ganoderma species DNA was used in PCR amplification with these primer pairs. Three specific restriction enzyme sites were identified in the ITS and intergenic spacer (IGS1) regions. The restriction enzymes MluI, SacI and HinfI were used to digest the ITS-PCR product and restriction enzymes TfiI, ScaI and HincII were used to digest the IGS1-PCR product. Of the three restriction enzymes used in each rDNA region, MluI specifically digested the ITS regions, and TfiI specifically digested the IGS1 region of oil palm Ganoderma. Analysis of the published ITS nucleotide sequences of 31 Ganoderma species showed that the MluI restriction site was not present in other Ganoderma species. The use of both specific primers and restriction enzyme analysis can be applied as a standard protocol to identify pathogenic Ganoderma in oil palm. In this study, the use of specific primers and PCR-RFLP analyses of the rDNA gave consistent results for the characterisation of pathogenic Ganoderma, and indicated that Ganoderma strains associated with BSR disease in oil palms belong to a single species. PMID- 15750750 TI - Research on basal stem rot (BSR) of ornamental palms caused by basidiospores from Ganoderma boninense. AB - Basidiospores were isolated from the fruiting bodies of Ganoderma infecting oil palms from an estate in Johor and from ornamental palms (including oil palms) from Singapore. The spores were then germinated to obtain homokaryotic mycelia. Based on clamp connection formation in paired hyphal fusions, tester strains were identified from the homokaryons isolated. Compatibility tests were then carried out using these testers to determine the relatedness of the homokaryotic Ganoderma isolates, both from Johor and from Singapore. Results from the compatibility tests showed that Ganoderma from both locations belong to the same species, while the Ganoderma isolates from Singapore share some common alleles. The pathogenicity tests carried out on Chrysalidocarpus lutescens seedlings using inoculum growing on rubber wood blocks showed that dikaryotic mycelia can cause basal stem rot infection. PMID- 15750751 TI - Treatment of hepatitis C virus infection--is there progress? PMID- 15750752 TI - Nasal carriage of Staphylococcus aureus and prevention of nosocomial infections. AB - This review summarizes the clinically relevant aspects of nasal carriage of Staphylococcus aureus. The epidemiology, associated risk, and the effects of eradication are discussed. The main conclusions are that nasal carriage of S. aureus is a well-defined risk factor for subsequent infection in nearly all categories of hospitalized patients that have been studied. However, studies that have been performed to evaluate the effect of eradication of carriage using mupirocin nasal ointment have been inconclusive so far in most subgroups. Only in patients on hemodialysis or chronic ambulatory peritoneal dialysis (CAPD) was a significant reduction of the infection rate found. But prolonged treatment in these groups carries a risk for the development of resistance. In surgical patients two randomized studies have found an effect on the surgical site infection rate in carriers that, when those studies are combined, was close to being statistically significant (p = 0.06). In non-surgical patients a significant delay in the onset of infection was found but the overall infection rate was not significantly different. When the results of all well-designed studies that have been performed are combined, a significant reduction of the nosocomial S. aureus infections in carriers is found (approximately 50% lower). Future studies should focus on treating carriers only and consider other treatment regimens. PMID- 15750753 TI - Prevalence of hepatitis C virus infection in different population groups in southern Italy. AB - BACKGROUND: A cross-sectional investigation was carried out between 2000 and 2002 to assess the prevalence of hepatitis C virus infection (HCV) in Naples, southern Italy. PATIENTS AND METHODS: Five groups of individuals were investigated, two at low risk and three at high risk for HCV infection. Blood sample sera were collected among 5,391 individuals (4,059 men and 1,332 women): 1,972 general practitioner (GP) patients and 781 employees of the National Cancer Institute (NCI) of Naples (low-risk groups); 524 male prisoners, 1,436 intravenous drug users (IDUs) and 678 hemodialysis patients (high-risk groups). RESULTS: Overall HCV seropositivity rates ranged from 6.4% among employees of the NCI to 37.4% among male prisoners. HCV infection tended to generally increase with age, but in IDUs and in male prisoners the upward trend leveled off at 50 years of age. As compared to GP patients, IDUs (both sexes) and male prisoners had a nearly 6-fold increased risk of HCV infection, while HCV was nearly 3-fold more common among hemodialysis patients. Employees of NCI were at reduced risk of HCV infection, particularly women (odds ratio = 0.3). CONCLUSION: The study findings confirmed the high risk for HCV infection in IDUs and identified other population groups in southern Italy that should be offered HCV screening and counselling given the severe implications of HCV infection on health. PMID- 15750754 TI - The role of Bordetella infections in patients with acute exacerbation of chronic bronchitis. AB - BACKGROUND: Acute exacerbations of chronic bronchitis (AECB) are associated with a variety of viral and bacterial infectious agents, some of which are potentially preventable by immunization. Bordetella pertussis, which causes whooping cough, has not been studied in this context. We aimed to assess the role of Bordetella infections in patients with AECB. PATIENTS AND METHODS: Patients with AECB, who presented to participating private practices in Basel, Switzerland, between October 2000 and June 2002, were evaluated by a standardized questionnaire, nasopharyngeal swabs for culture (Bordetella spp.), and PCR (Bordetella spp. and selected other respiratory pathogens) and paired blood samples for serologic diagnosis of Bordetella infection. RESULTS: A total of 26 patients (34-86 years of age) were recruited. All culture and PCR samples were negative. Serology revealed Bordetella infection in eight (31%) patients. Duration of cough was shorter in patients with Bordetella infection compared to those without Bordetella infection (mean 15 days vs 41 days, p = 0.04). Cough > or = 21 days duration was present in three (43%) of seven patients with evidence of Bordetella infection compared to 17 (94%) of 18 controls (p = 0.012). Progression to convalescence from initial to follow-up visit after 4-6 weeks was comparable between both groups. CONCLUSION: Bordetella infections appear to play a significant role in AECB and preventive measurements such as immunization with acellular pertussis vaccines should be considered. Extended investigations are necessary to confirm our preliminary and provocative findings. PMID- 15750755 TI - Incidence of respiratory syncytial virus positivity in young Italian children referred to the emergency departments for lower respiratory tract infection over two consecutive epidemic seasons. AB - BACKGROUND: The rate and severity of respiratory syncytial virus (RSV) infections within the same nation may vary from one year to another. PATIENTS AND METHODS: The "Osservatorio VRS" study collected epidemiological data on RSV infection among Italian children aged < or = 4 years referred to emergency departments of 14 centers, for suspected lower respiratory tract infections (LRTI) in two consecutive RSV seasons (October 2000-April 2001 and October 2001-April 2002). Medical history and physical examination were recorded and an immunoenzymatic RSV test was performed on nasal secretions. Study variables were collected and evaluated separately, then compared. RESULTS: In all, 272 and 756 children were included in the two seasons, respectively, of which 31.6% and 19.2% were RSV positive (+). Children of the first season had lower gestational and chronological age, higher rates of chronic lung disease (CLD), very low birth weight (< 1,500 g), larger use of corticosteroids or bronchodilators. Main risk factors for RSV infection were a young age (< 1 year) and a low birth weight (< 1,500 g). RSV infection reached its peak in February (first season) or March (second season), with an earlier appearance in the northern and central as compared to the southern regions. Rate of hospitalization and LRTI was higher in RSV+ children, especially if young. CONCLUSION: Although rhythms of the RSV seasons and patient characteristics may vary from one year to another, the severity of RSV disease in nonprophylaxed infants and young children remains high. PMID- 15750756 TI - Interferon alpha-2a plus ribavirin 1,000/1,200 mg versus interferon alpha-2a plus ribavirin 600 mg for chronic hepatitis C infection in patients on opiate maintenance treatment: an open-label randomized multicenter trial. AB - BACKGROUND: Many intravenous opiate users are infected with hepatitis C virus (HCV) but few are treated. Although this complies with various guidelines, virtually no published evidence supports such a recommendation. PATIENTS AND METHODS: In a multicenter study, HCV-infected patients in opiate maintenance treatment programs received interferon plus high- or low-dose ribavirin (1,000/1,200 mg or 600 mg). HIV-coinfected patients were not included. Endpoints were feasibility, efficacy, side effects, and reasons for dropout. RESULTS: Of the 420 patients who tested positive for HCV, 27 (6%) were enrolled; 393 (94%) either failed to meet the inclusion criteria or refused treatment. Virologic end of-treatment response was achieved in 12/27 patients, and sustained response in 13/27 (48%). Response depended on viral genotype, not ribavirin dose. The two doses of ribavirin did not differ in their side effects. CONCLUSION: In a small fraction of HCV-infected intravenous drug users in an opiate maintenance treatment program, antiviral therapy was feasible, safe, and effective. The success rate was comparable to that achieved in controlled studies that excluded drug users. PMID- 15750757 TI - Peginterferon alfa-2b treatment for patients affected by acute hepatitis C: presentation of six case reports. AB - Acute hepatitis C often progresses to chronic infection (70%). In this clinical study, we evaluated if early treatment with peginterferon alfa-2b can prevent acute hepatitis C from developing into a chronic disease. Six patients with acute hepatitis C, based on a well-documented hepatitis C virus (HCV) seroconversion with high alanine aminotransferase (ALT) levels (> 10 x ULN) and persistent HCV RNA titers after 3 months from disease onset, were consecutively treated with peginterferon alfa-2b at 1.5 microg/kg/weekly/sc for 24 weeks. The viral load was quantified by PCR assay. Response was defined as undetectable HCV RNA and normal ALT levels at the end of therapy and after a 6-month follow-up. All patients completed therapy; at the end of therapy, 5/6 patients (83%) responded and no relapses were observed during follow-up. No correlation was found between treatment response and pretreatment viral load, viral genotype, and interval between acute infection diagnosis and start of therapy. PMID- 15750758 TI - Fulminant cerebral malaria in a Swiss patient. AB - Malaria remains the most important parasitic disease worldwide. Falciparum malaria is a medical emergency and requires immediate diagnosis and treatment. Cerebral malaria is a rapidly progressive, potentially fatal complication of Plasmodium falciparum infection. This case, including post-mortem observations, histology, and laboratory diagnosis, emphasizes the necessity of appropriate advice regarding malaria prophylaxis before travel to an endemic area. Malaria should always be considered in the differential diagnosis of patients presenting with fever and/or nonspecific flu-like symptoms after traveling to endemic countries. PMID- 15750759 TI - Brain abscesses complicating Staphylococcus aureus sepsis in a premature infant. AB - Brain abscess is a rare complication of staphylococcal bacteremia in infants. Here we present a case of a premature infant who developed multiple brain abscesses 12 weeks following an episode of inadequately treated Staphylococcus aureus sepsis. The abscess developed in the absence of trauma, prior surgery, cyanotic heart disease, or immune defect. The initial staphylococcal isolate exhibited identical pulsed-field gel electrophoresis pattern with that of the isolate cultured from abscess aspirate. The infant was successfully treated by surgical drainage and administration of antibiotics for 12 weeks, initially teicoplanin and meropenem followed by trimethoprim/sulfamethoxazole, without neurological or developmental sequelae. Staphylococcal bacteremia in neonates should be vigorously treated to prevent life-threatening complications. PMID- 15750760 TI - Deadly carousel or difficult interpretation of new diagnostic tools for Whipple's disease: case report and review of the literature. AB - Whipple's disease is a rare systemic disorder classically presenting with weight loss, arthralgias, and diarrhea, which was first described in 1907. The causative bacterium Tropheryma whipplei, is a fastidious organism not growing on conventional media. Before the introduction of polymerase chain reaction (PCR) based methods, the diagnostic gold standard was histological detection of diastase-resistant periodic acid Schiff (PAS)-positive macrophages or electron microscopy. As in the present case, contradictory results between the former and new diagnostic methods may obscure the correct diagnosis. We critically summarize the performance of the different diagnostic methods and discuss their impact on the clinical management of patients with suspected Whipple's disease. PMID- 15750761 TI - Immune reconstitution inflammatory syndrome (IRIS)--another new disease entity following treatment initiation of HIV infection. AB - After the introduction of highly active antiretroviral therapy (HAART) in 1995 it took 3 years to recognize the new syndrome of lipodystrophy as a long-term complication of HAART [1]. It was found to be a direct toxic consequence of the various drugs used in combination regimens. Another 2 years later, an additional syndrome was described in patients who recently initiated HAART-the immune reconstitution inflammatory syndrome (IRIS) [2-4]. Although already recognized in the pre-HIV era in patients with treatment for tuberculosis [5, 6] and leprosy [7], it became substantially more frequent in HIV-infected patients on HAART. The term IRIS describes a collection of different inflammatory disorders which are associated with paradoxical deterioration of various preexisting infectious processes following commencement of HAART in HIV-infected patients. These preexisting infections in individuals with IRIS may have been previously diagnosed and treated or may have been symptomless and later revealed by the patient's improved capacity to mount an inflammatory response. PMID- 15750762 TI - Molecular basis of immunoglobulin variable region gene usage in systemic autoimmunity. AB - This review focuses on the immunoglobulin variable region (IgV) chain gene usage in patients with systemic autoimmune diseases, with particular emphasis on systemic lupus erythematosus (SLE), a condition known to be associated with the production of a number of characteristic autoantibodies as an abnormality. The IgV repertoire is shaped by a variety of molecular and selective influences that are difficult to distinguish. Studies of IgV gene rearrangement by PCR of individual B cells permitted insight in the differential impact of these processes, including somatic hypermutation and indications of receptor editing/revision. Although the majority of current data indicate that there is no major molecular abnormality in V(D)J recombination in SLE patients, abnormalities in subsequent events, such as the degree of receptor editing and somatic hypermutation, and positive and negative selection fundamentally alter the composition of the peripheral B-cell repertoire. Identification of the mechanisms that influence the IgV gene and B-cell repertoire may allow new therapeutic approaches in autoimmunity. PMID- 15750763 TI - Effects of cilostazol on lipid and fatty acid metabolism. AB - Cilostazol is a selective inhibitor of phosphodiesterase III with anti-platelet aggregatory and vasodilating properties. Randomised, double-blind, placebo controlled trials in 2702 patients with intermittent claudication demonstrated that cilostazol significantly increased walking distances compared with placebo. Furthermore, the agent has beneficial effects on the serum lipid profile and fatty acid composition in plasma. Consequently, cilostazol may be useful to prevent atherosclerosis from progressing by ameliorating lipid and fatty acid metabolism. PMID- 15750764 TI - Blockade of the Ras pathway by manumycin, a farnesyltransferase inhibitor, overcomes the resistance of myeloma plasma cells to Fas-induced apoptosis. AB - Ras activation (by point mutation or binding of IL-6) is frequently observed in multiple myeloma (MM). As farnesylation of Ras protein by farnesyltransferase is a critical step for Ras functional activity, farnesyltransferase inhibitors (FTI) have emerged as potential anti-cancer agents. Manumycin, a natural FTI, prevents proliferation and induces apoptosis of myeloma cells refractory to Fasand drug induced cell death. Fas pathway analysis showed that Fas-resistant apoptosis of Fas-positive myeloma cells parallels FLIP (FLICE/caspase-8-inhibitory protein) expression. Treatment of fresh purified myeloma cells, myeloma cell clone-2 and U266 cell line with manumycin induced down-regulation of FLIP expression with concomitant expression of Apo 2.7 antigen, the marker of early apoptosis. Down regulation of FLIP mRNA levels in drug-treated cells was associated to suppression of the transcription factor NF-kappaB that plays a central role in chemoresistance, survival and proliferation of myeloma cells. Further analysis showed that manumycin-induced apoptosis involved caspases activation and was prevented by the addition of caspases specific inhibitors. Finally, pretreatment of Fas-resistant/FLIP-positive cells with manumycin sensitised them to Fas triggered apoptosis. Overall results indicate that manumycin-induced apoptosis involves Fas pathway. FTIs may thus be proposed as a promising class of anti cancer agents which can boost the cytotoxic effect of conventional drugs by overcoming NF-kappaB activation and Fas-resistant apoptosis. PMID- 15750765 TI - Changes in lipid metabolism in women with age-related macular degeneration. AB - Age-related macular degeneration (AMD) is one of the leading causes of visual loss among people aged 65 and older. At present the origin of AMD still remains unknown. The objective was to evaluate the chosen lipid and lipoprotein concentrations in blood of patients with AMD. Sixty women aged 55-71 (mean age 65.1+/-5.7) were treated in the outpatient ophthalmological clinic for more than two years because of AMD. We evaluated total serum cholesterol (TCH), triglycerides (TG), HDL-cholesterol (HDL), LDL-cholesterol (LDL), lipoprotein (a) (Lp(a)), apolipoprotein AI (Apo AI) and apolipoprotein B (Apo B) by direct spectrophotometry (Human and Randox standard kits, USA). We found a significant increase of TCH, LDL and TG (224.36+/-41.67 mg/dl, 159.02+/-39.66 mg/dl and 120.92+/-42.64 mg/dl), and a significant decrease of HDL (38.68+/-6.36 mg/dl) in the AMD patients when compared with the control group. We have not found a significant difference in the average TG level between the studied groups. The concentration of Apo B was markedly increased (164.66+/-46.46 mg/dl) and Apo AI concentration was markedly decreased (128.9+/-17.01 mg/dl) in the AMD patients when compared with the control group. There was no significant difference in the concentration of the Lp(a) between the two groups. The results of our present study could point to the fact that changes in the lipid metabolism could be one of the very important risk factors involved in the pathogenesis of AMD. PMID- 15750766 TI - Relationship between dose of methotrexate, apoptosis, p53/p21 expression and intestinal crypt proliferation in the rat. AB - Previous studies have shown that apoptosis is induced by cytotoxic chemotherapy and precedes hypoproliferation of intestinal crypt cells. However, the relationship between the degree of intestinal apoptosis and crypt cell hypoproliferation may not be directly related. The purpose of this study was to investigate the relationship between apoptosis and hypoproliferation with increasing doses of chemotherapy. Eleven groups of breast cancer-bearing DA rats were treated with two doses of methotrexate (MTX) i. m. at varying concentrations (0.5, 1.5, 2.5 and 5.0 mg/kg) or saline (control). Animals were killed at 6 or 24 h following treatment. The small and large intestines were examined for apoptosis, villous area (small intestine), crypt length and mitotic count per crypt. Immunohistochemical expression of p53 and p21(waf1/cip1) (p21) were examined quantitatively. Data were analysed using Peritz' F-test. Low dose MTX (0.5 mg/kg) did not change p53 expression at 6 h but induced a 15-fold increase in apoptosis in the crypts of the small intestine. This was associated with only a minor reduction in crypt cell proliferation. Higher doses of MTX increased p53 expression and caused a lower (7-fold) but more prolonged peak of apoptosis that was accompanied by reduced villous area, shortened crypts and a more profound reduction in crypt cell proliferation. Unlike the small intestine, apoptosis in the colon was 10-fold lower, proportional to the dose of MTX and did not induce overt damage. Expression of p21 did not change with any dose at either timepoint. We conclude that apoptosis is not always associated with crypt cell hypoproliferation and that the small intestine can recover after low dose MTX despite a heightened peak of apoptosis of crypt cells. PMID- 15750767 TI - Rapid detection and differentiation of Gram-negative and Gram-positive pathogenic bacteria in urine using TaqMan probe. AB - Urinary tract infection has been shown to be quite complicated and often difficult to diagnose and treat. For appropriate diagnosis, it is very important to find the correct Gram stain classification as soon as possible, especially in severe cases where there is a possibility of severe sepsis developing. In order to solve this problem, we developed a new method to detect a Gram stain of bacteria obtained from 1 ml of urine from urinary tract infection patients using a consensus real-time PCR protocol with a TaqMan probe that allows detection of spiked bacterial 16S DNA from urine. We extracted DNA of 55 urine samples obtained from patients with complicated urinary tract infection and at the same time performed urine culture testing. After DNA extraction, they were subjected to real-time PCR using a TaqMan discrimination system. Sixteen kinds of bacteria were cultured from the urine culture testing. Of these bacteria, eight were classified as Gram-positive bacteria and the other eight were classified as Gram negative bacteria. Of the 55 samples, the TaqMan technique result showed 27 samples that were classified as Gram-negative bacteria; 11 samples that were Gram positive, 10 that included both Gram-negative and -positive bacteria, and 7 that showed no amplification. The classifications of all samples corresponded exactly to those determined by urine culture testing. The present genotyping method of real-time PCR using a TaqMan discrimination system could be applied to the rapid detection of Gram-positive or -negative bacteria in urine of urinary tract infection patients. This assay can differentiate those species tested, but whether the presence of other (untested) bacteria could lead to misinterpretation is unknown. For further investigation, it is important to test other (untested) bacteria in the near future. PMID- 15750768 TI - Distribution of C-->T and T-->C polymorphisms of the urokinase-type plasminogen activator gene in children with type 1 diabetes mellitus and insulin resistance. AB - We analysed the distribution of genotypes and frequency of alleles of two polymorphisms in the urokinase-type plasminogen activator (uPA) gene: a C-->T substitution in exon 6 and a T-->C substitution in intron 7 in 89 children with type 1 diabetes mellitus and insulin resistance compared with 120 non-diabetic control subjects. All genotypes were determined by the allele-specific polymerase chain reaction. We found that the frequency of the T/T homozygote (15%) in the patient group was significantly (P<0.05) higher than in the controls (7%). There were no differences in the distribution of the T-->C polymorphism between patients and controls, which suggests that this genetic change is probably phenotypically silent. In conclusion, our results indicate that the higher percentage of T/T homozygotes in patients might be associated with T1DM coexisting with insulin resistance. PMID- 15750769 TI - Mercury levels and relationships in water, sediment, and fish tissue in the Willamette Basin, Oregon. AB - In Oregon's Willamette River Basin (the Basin), health advisories currently limit consumption of fish that have accumulated methylmercury (MeHg) to levels posing a significant human health risk. These advisories created the requirement for a mercury total maximum daily load for the Basin, which required a greater understanding of the behavior, distribution, and levels of mercury and MeHg in the Basin. In 2002, the Oregon Department of Environmental Quality initiated a study to measure (using ultraclean techniques) mercury and MeHg levels in water, sediment, and fish samples collected throughout the Basin. Results from the Middle Fork (nominal background) suggested that naturally occurring surface-water concentrations of mercury and MeHg would on an annual average basis be expected in the range of 0.5 to 1.0 and 0.04 to 0.06 ng L(-1), respectively. Concentrations in the Coast Fork (Cottage Grove), which were markedly higher, are likely the result of historical mining discharges. The possibility exists that wetlands alone could contribute the dissolved MeHg levels (approximately 0.04 ng L(-1)) observed in the Main Stem. Mercury levels in sediment were similar, and near background, in the Main Stem, Coast Fork (Row River), and Middle Fork but significantly increased in the Coast Fork (Cottage Grove). Fish tissue mercury levels were typically highest in piscivorous and lowest in invertivorous species but highest in the Coast Fork (Cottage Grove). In the Coast Fork and Cottage Grove Reservoir, discharges from historical mercury mining activities appear to have significantly impacted water, sediment, and fish tissue levels; however these impacts do not appear to extend into the Main Stem. Basinwide mercury data are at present too spottily distributed to determine whether significant mercury point sources exist along the Main Stem. PMID- 15750770 TI - Monitoring of antifouling booster biocides in water and sediment from the port of Osaka, Japan. AB - Concentrations of booster antifouling compounds in the port of Osaka, Japan were assessed. Concentrations of Sea-Nine 211 (4,5-dichloro-2-n-octyl-3 isothiazolone), thiabendazole (2-(4-thiazolyl)-benzimidazole), IPBC (3-iodo-2 propynyl butylcarbamate), Diuron (3,4-dichlorophenyl-N, N-dimethylurea), Irgarol 1051 (2-methylthio-4-t-butylamino-6-cyclopropylamino-s-triazine), and M1 (2 methylthio-4-t-butylamino-6-amino-s-triazine) in port water samples were in the range of <0.003-0.004 microg L(-1), <0.0008-0.020 microg L(-1), <0.0007-1.54 microg L(-1), <0.0008-0.267 microg L(-1), and <0.0019-0.167 microg L(-1), respectively. IPBC was not detected in the water samples, but the concentration of Diuron was higher than any previously reported. The concentrations of Sea-Nine 211, thiabendazole, Diuron, Irgarol 1051, and M1 in sediment samples were in the range of <0.04-2.4 microg kg(-1) dry, <0.08-1.2 microg kg(-1) dry, <0.64-1350 microg kg(-1) dry, <0.08-8.2 microg kg(-1) dry, and <0.18-2.9 microg kg(-1) dry, respectively. IPBC was again not detected. The levels of Sea-Nine 211, Diuron, and Irgarol 1051 in water and sediment samples were high in a poorly flushed mooring area for small and medium-hull vessels. Levels of Diuron and Irgarol 1051 were highest in summer. The concentration of Sea-Nine 211 in water increased between August and October 2002. Except for M1, increases in the levels of booster biocides in sediment were observed during the study period. The sediment water partition (Kd) was calculated by dividing the concentrations in sediment by the concentrations in water. The Kd values for Sea-Nine 211, thiabendazole, Diuron, Irgarol 1051, and M1 were 690, 180, 2700, 300, and 870. The Kd value for these alternative compounds was lower than for TBT. PMID- 15750771 TI - Tissue-specific cadmium and metallothionein levels in rainbow trout chronically acclimated to waterborne or dietary cadmium. AB - Rainbow trout were exposed to a sublethal concentration of waterborne Cd (0 or 3 microg/L) or dietary Cd (0 or 500 mg/kg dry wt) for 30 days to induce acclimation, and tissue Cd and metallothionein (MT) levels were examined. The greatest Cd concentrations were observed in the kidney followed by the gills and liver of the fish exposed to Cd via water, but in the gut tissues followed by the kidney, liver, and gills for dietary-exposed fish, reflecting a variation depending on the route of Cd exposure. Some MT was found in the nonacclimated naive fish with no experience of elevated Cd exposure, and these background MT levels were quite high in the posterior intestine (480 microg/g), cecae (257 microg/g), and liver (248 microg/g) relative to other tissues (7-50 microg/g). With exposure to both waterborne and dietary Cd, MT levels rose significantly in all observed tissues. The increases relative to the control levels of MT in naive fish were in the order: kidney (5.4 times) > gills (4.6) > liver (1.3) for the waterborne exposure group, and in the order kidney (19.3 times) >> cecae and posterior intestine (approximately 6.5 times) > liver and stomach (approximately 5 times) > midintestine (4.3 times) > gills (2.1 times) for the dietary exposure group. At 24 hours after an acute gastrointestinal dose of Cd (276 microg/kg) infused into the stomach of dietary exposure groups, large increases of total Cd but not MT levels were found in the gut tissues of nonacclimated fish; in the Cd acclimated fish, the posterior intestine was greatly affected with decreases in Cd (71%), Zn (33%), Cu (70%) and MT (46%) levels, suggesting an enhanced sloughing of tissue materials after infusion. Exposure to Cd did not cause any notable decrease of Zn or Cu in any tissue, except that found in the posterior intestine. However, a molar analysis indicated that although Cd levels remained less than MT binding capacity in both waterborne and dietary exposure groups, the total metal levels (Cd + Zn + Cu) greatly exceeded MT binding capacity in all tissues of Cd-exposed fish, suggesting a potential competition of Cd with other metals for binding sites on MT and non-MT proteins in the tissues. PMID- 15750772 TI - Microcosm evaluation of the toxicity and risk to aquatic macrophytes from perfluorooctane sulfonic acid. AB - Perfluorooctane sulfonic acid (PFOS) is an anthropogenic contaminant detected in various environmental and biologic matrices. This compound is a fluorinated surfactant, a class of molecules renowned for their persistence and their global distribution but for which few ecotoxicological data are currently available, especially under field conditions. The toxicity of PFOS to the aquatic macrophytes Myriophyllum sibiricum and M. spicatum was investigated using 12,000 L outdoor microcosms. Replicate microcosms (n = 3) were treated with 0.3, 3, 10, and 30 mg/L PFOS as the potassium salt and assessed at regular intervals during a period of 42 days. M. sibiricum was more sensitive to PFOS under these simulated field conditions than M. spicatum. Toxicity was observed in the evaluated end points at > 3 mg/L PFOS for EC10s and > 12 mg/L PFOS for EC50s for M. spicatum and in M. sibiricum at > 0.1 mg/L PFOS for EC10s and > 1.6 mg/L PFOS for EC50s. The no observed-effect concentration (NOEC) for M. spicatum was consistently > or = 11.4 mg/L PFOS, whereas the NOEC for M. sibiricum was > or = 0.3 mg/L PFOS. A risk assessment for these plants estimated a negligible probability of toxicity being observed in these plants from PFOS exposure at current environmental concentrations. PMID- 15750773 TI - Accumulation of selenium and lack of severe effects on productivity of American dippers (Cinclus mexicanus) and spotted sandpipers (Actitis macularia). AB - Selenium has been found at elevated concentrations in water, sediments, and aquatic biota in the Elk River (British Columbia, Canada) and some of its tributaries downstream of several coal mines. Selenium water concentrations in those areas exceed Canadian and British Columbia guidelines and are above levels at which adverse effects to fish and waterfowl could occur. We compared selenium concentrations in the eggs of two riverine waterbirds, American dippers and spotted sandpipers, with measures of productivity: the number of eggs laid, egg hatchability, and nestling survival. In American dippers, the mean egg selenium concentration from the exposed areas, 1.10 +/- SE 0.059 microg/g wet weight, was indistinguishable from the reference areas, 0.96 +/- SE 0.059 microg/g wet weight. For spotted sandpipers, the mean egg selenium concentration in the exposed areas, 2.2 +/- 0.5 microg/g wet weight, was significantly higher than in the reference areas, 1.2 +/- 0.14 microg/g wet weight, but less than reported thresholds for waterfowl and other shorebirds. There were no significant differences in egg hatchability between dippers in reference and exposed areas, but reduced hatchability was apparent for sandpipers in exposed locations. Despite the slightly reduced hatchability in sandpipers, overall productivity was higher than regional norms for both species; thus, selenium did not affect the number of young recruited to local populations. We did not observe teratogenic effects in either species, although none was expected at these concentrations. Despite moderately high selenium concentrations in the water, mean egg selenium concentrations were less than predicted from uptake models. We hypothesise that the relatively low uptake of selenium into the eggs of the two waterbirds in this study is likely due to their lotic environment's low biological transformation and uptake rates. PMID- 15750774 TI - Biodegradation of the insecticide N,N-diethyl-m-toluamide by fungi: identification and toxicity of metabolites. AB - Fungi (Cunninghamella elegans ATCC 9245, Mucor ramannianus R-56, Aspergillus niger VKMF-1119, and Phanerochaete chrysosporium BKMF-1767) were tested to elucidate the biologic fate of the topical insect repellent N,N-diethyl-m toluamide (DEET). The elution profile obtained from analysis by high-pressure liquid chromatography equipped with a reverse-phase C-18 column, showed that three peaks occurred after incubation of C. elegans, with which 1 mM DEET was combined as a final concentration. The peaks were not detected in the control experiments with either DEET alone or tested fungus alone. The metabolites produced by C. elegans exhibited a molecular mass of 207 with a fragment ion (m/z) at 135, a molecular mass of 179 with an m/z at 135, and a molecular mass of 163 with an m/z at 119, all of which correspond to N,N-diethyl-m-toluamide-N oxide, N-ethyl-m-toluamide-N-oxide, and N-ethyl-m-toluamide, respectively. M. ramannianus R-56 also produced N, N-diethyl-m-toluamide-N-oxide and N-ethyl-m toluamide but did not produce N-ethyl-m-toluamide-N-oxide. For the biologic toxicity test with DEET and its metabolites, the freshwater zooplankton Daphnia magna was used. The biologic sensitivity in decreasing order was DEET > N-ethyl-m toluamide > N,N-diethyl-m-toluamide-N-oxide. Although DEET and its fungal metabolites showed relatively low mortality compared with other insecticides, the toxicity was increased at longer exposure periods. These are the first reports of the metabolism of DEET by fungi and of the biologic toxicity of DEET and its fungal metabolites to the freshwater zooplankton D. magna. PMID- 15750775 TI - Environmental toxicity studies using chickens as surrogates for wildlife: effects of injection day. AB - Domestic chicken embryos are frequently used for avian developmental toxicity studies of polyhalogenated aromatic hydrocarbons, which are often injected into eggs with oil-based vehicles. Injection times range from immediately prior to incubation (embryonic day zero, E0) to after 4 days of incubation (E4) and beyond. Because the majority of organogenesis in chicken embryos occurs during the first 4 days of development, injection after E0 may miss critical, sensitive, developmental periods. We evaluated whether differences in the day of vehicle administration would lead to differences in standard measures of embryotoxicity. We assessed embryotoxicity using mortality, organ somatic indices, teratogenesis, and behavior in hatchling chickens developmentally exposed to a high volume (1.0 microl/g egg) of corn oil vehicle, which was injected into the airsac at E0 or E4. The E0 vehicle group had 76.5% higher overall embryo mortality, embryos died earlier in development, and hatchlings took more than two times longer to right in a righting reflex test compared to the E4 vehicle group. Other behavioral results demonstrated that hatchling chickens from the E0 vehicle group performed differently from their respective no-inject controls, whereas hatchling chickens from the E4 vehicle group did not. The bursal somatic index differed statistically by injection day and weighed 23.7% more in the E0 vehicle group than the E4 vehicle group. These results suggest that the embryonic day of contaminant injection is an important consideration, particularly when using a high volume of vehicle to evaluate developmental toxicity of a contaminant on embryo mortality or behavior. PMID- 15750776 TI - Comparative aquatic toxicity evaluation of 2-(thiocyanomethylthio)benzothiazole and selected degradation products using Ceriodaphnia dubia. AB - 2-(Thiocyanomethylthio)benzothiazole (TCMTB) is a biocide used in the leather, pulp and paper, and water-treatment industries. TCMTB may enter aquatic ecosystems during its manufacture and use. TCMTB is environmentally unstable; therefore, it is important to evaluate the toxicity of the more persistent degradation products. This study compared the toxicity of TCMTB with its degradation products 2-mercaptobenzothiazole (2-MBT), 2-(methylthio)benzothiazole (MTBT), benzothiazole (BT), and 2-hydroxybenzothiazole (HOBT). Toxicity was determined using Ceriodaphnia dubia 48-hour acute and 7-day chronic test protocols. TCMTB was the most toxic compound evaluated in both the acute and chronic tests with EC50s of 15.3 and 9.64 microg/L, respectively. 2-MBT, the first degradation product, was the second most toxic compound with acute and chronic EC50s of 4.19 and 1.25 mg/L, respectively. The toxicity of MTBT and HOBT were similar with acute EC50s of 12.7 and 15.1 mg/L and chronic EC50s of 6.36 and 8.31 mg/L, respectively. The least toxic compound was BT with acute and chronic EC50s of 24.6 and 54.9 mg/L, respectively. TCMTB was orders of magnitude more toxic than its degradation products. Toxicity data on these benzothiazole degradation products is important because of concerns regarding their release, degradation, persistence, and non-target organism effects in aquatic ecosystems. PMID- 15750777 TI - Assessing contaminant sensitivity of endangered and threatened aquatic species: part III. Effluent toxicity tests. AB - Toxicity tests using standard effluent test procedures described by the U.S. Environmental Protection Agency were conducted with Ceriodaphnia dubia, fathead minnows (Pimephales promelas), and seven threatened and endangered (listed) fish species from four families: (1) Acipenseridae: shortnose sturgeon (Acipenser brevirostrum); (2) Catostomidae; razorback sucker (Xyrauchen texanus); (3) Cyprinidae: bonytail chub (Gila elegans), Cape Fear shiner (Notropis mekistocholas) Colorado pikeminnow (Ptychocheilus lucius), and spotfin chub (Cyprinella monacha); and (4) Poecillidae: Gila topminnow (Poeciliopsis occidentalis). We conducted 7-day survival and growth studies with embryo-larval fathead minnows and analogous exposures using the listed species. Survival and reproduction were also determined with C. dubia. Tests were conducted with carbaryl, ammonia--or a simulated effluent complex mixture of carbaryl, copper, 4 nonylphenol, pentachlorophenol and permethrin at equitoxic proportions. In addition, Cape Fear shiners and spotfin chub were tested using diazinon, copper, and chlorine. Toxicity tests were also conducted with field-collected effluents from domestic or industrial facilities. Bonytail chub and razorback suckers were tested with effluents collected in Arizona whereas effluent samples collected from North Carolina were tested with Cape Fear shiner, spotfin chub, and shortnose sturgeon. The fathead minnow 7-day effluent test was often a reliable estimator of toxic effects to the listed fishes. However, in 21 % of the tests, a listed species was more sensitive than fathead minnows. More sensitive species results varied by test so that usually no species was always more or less sensitive than fathead minnows. Only the Gila topminnow was consistently less sensitive than the fathead minnow. Listed fish species were protected 96% of the time when results for both fathead minnows and C. dubia were considered, thus reinforcing the value of standard whole-effluent toxicity tests using those two species. If the responses of specific listed species are important for management decisions, our study supports the value in developing culture and testing procedures for those species. PMID- 15750778 TI - Thyroid lesions and dioxin accumulation in the livers of jungle crows (Corvus macrorhynchos) in urban and suburban Tokyo. AB - Wild jungle crows (Corvus macrorhynchos) captured from three different areas of Tokyo were examined to evaluate environmental contamination of dioxins. In addition to the pathologic examination of their whole body, accumulation of dioxins, mRNA expression of the aryl hydrocarbon receptor (AhR), and pentoxyresorufin-O-depenthylase (PROD) activity in the liver were determined. Marked histopathologic changes were observed in the thyroid glands, especially in the crows from the urban downtown area. Levels of dioxins and their toxic equivalents (TEQs) and AhR mRNA expression in the livers of the crows from the urban area were higher than those from the suburban area. There was a high correlation between the levels of TEQs and PROD activity. The results of the present study demonstrated that jungle crows possess AhR-mediated toxicologic pathways similar to those of mammals and suggest the possibility that the thyroidal changes observed in the adult crows from the urban areas are one of the toxic manifestations resulting from exposure to dioxins and other environmental chemicals. PMID- 15750779 TI - Environmental toxicity studies using chickens as surrogates for wildlife: effects of vehicle volume. AB - Domestic chicken embryos are frequently used for avian developmental toxicity studies of polyhalogenated aromatic hydrocarbons, which are often injected into eggs with oil-based vehicles. The volume of toxicant and vehicle injected ranges from relatively low volumes (0.1-0.5 microl/g egg) to relatively high volumes (1.0 microl/g egg and above). Previous research from our laboratory suggested that high volumes of vehicle oil may disrupt normal growth of chicken embryos, possibly from hypoxia-like effects. This analysis explored the potential effects of vehicle volume on developing chicken embryos. We assessed standard measures of mortality, organ growth, body growth, and behavior from chickens developmentally exposed in ovo prior to incubation to low (0.1 microl/g egg) or high volumes (1.0 microl/g egg) of corn oil injected into airsacs or to no injection. The chickens receiving high volumes of oil showed increases in overall embryonic mortality and early embryo mortality compared to chickens receiving low volumes of oil or no injection. The chickens receiving high volumes of oil showed decreased activity during righting reflex, running time, visual discrimination, and olfactory aversion tests, and increased activity during an open-field activity test compared to chickens receiving low volumes of oil or no injection. Somatic endpoints do not appear to be affected by high volume injections. These results suggest that high volumes of vehicle injected into airsacs of eggs may lead to hypoxia-like conditions that increase embryonic mortality and disrupt simple behaviors. However, some effects of volume may diminish when injections are performed later in incubation. PMID- 15750780 TI - Comparative sublethal toxicity of nine pesticides on olfactory learning performances of the honeybee Apis mellifera. AB - Using a conditioned proboscis extension response (PER) assay, honeybees (Apis mellifera L.) can be trained to associate an odor stimulus with a sucrose reward. Previous studies have shown that observations of conditioned PER were of interest for assessing the behavioral effects of pesticides on the honeybee. In the present study, the effects of sublethal concentrations of nine pesticides on learning performances of worker bees subjected to the PER assay were estimated and compared. Pesticides were tested at three concentrations. The highest concentration of each pesticide corresponded to the median lethal dose value (48 h oral LD50), received per bee and per day, divided by 20. Reduced learning performances were observed for bees surviving treatment with fipronil, deltamethrin, endosulfan, and prochloraz. A lack of behavioral effects after treatment with lambda-cyalothrin, cypermethrin, tau-fluvalinate, triazamate, and dimethoate was recorded. No-observed-effect concentrations (NOECs) for the conditioned PER were derived for the studied pesticides. Our study shows that the PER assay can be used for estimating sublethal effects of pesticides on bees. Furthermore, comparisons of sensitivity as well as the estimation of NOECs, useful for regulatory purposes, are possible. PMID- 15750781 TI - Radiolytic degradation of the herbicide dicamba for environmental protection. AB - The radiolytic degradation of the widely used herbicide dicamba (3,6-dichloro-2 methoxybenzoic acid), employing gamma irradiation in laboratory batch conditions and with a beam of accelerated electrons in flow-through installation, was investigated. The effects of dose magnitude, ozone or hydrogen peroxide in irradiated solution, and scavengers such as nitrate and hydrogen carbonate on the effectiveness of dicamba decomposition and the products formed were investigated. Changes in the toxicity of irradiated solutions were measured with the Microtox and Spirotox toxicity tests. The application of radiolytic degradation was also examined for decomposition of herbicides in commercial agrochemical preparations. PMID- 15750782 TI - Efficacy of two wastewater treatment plants in removing genotoxins. AB - The genotoxic potential of influents and effluents of two different wastewater treatment plants (WTP-A and WTP-B) located in the Rouen, France, area was evaluated by the SOS chromotest without metabolic activation (on Escherichia coli PQ37) and the Ames fluctuation test (on Salmonella typhimurium strains TA 98, 100, TA 102) with and without metabolic activation. The wastewater samples were taken during two 1-week periods in January and April 2003. The simultaneous use of the SOS chromotest and Ames fluctuation test allowed us to evaluate the efficacy of the wastewater treatment plants at removing genotoxins. Genotoxins were detected with the Ames test but not with the SOS chromotest. Out of a total of 24 influents tested (14 for WTP-A and 10 for WTP-B), almost all were genotoxic in at least one Ames test strain (71% for WTP-A and 100% for WTP-B). In contrast, all of the tested effluents were nongenotoxic. This work showed that the treatment process used in the 2 wastewater treatment plants studied (activated sludge) was able to remove the genotoxins detected in their influents. Nevertheless, studies could be undertaken to determine which step of the treatment process removes genotoxins and whether WTP sludge use could be a source of genotoxic contamination for humans and the environment. PMID- 15750784 TI - Matrix metalloproteinase-2: mechanism and regulation of NF-kappaB-mediated activation and its role in cell motility and ECM-invasion. AB - Matrix metalloproteinases belong to a family of enzymes that degrade the extracellular matrix (ECM) components and play an important role in tissue repair, tumor invasion, and metastasis. ECM proteins, cytokines, and certain other factors regulate MMP activity. OPN, an ECM protein, has been found to be overexpressed in various cancers, and it has been shown to correlate with the metastatic potential. Although such reports indicate that OPN plays an important role in the ability of tumor cells to survive and metastasize to secondary sites, the mechanism by which OPN regulates these processes is yet to be understood. In this study we report that native purified human OPN can induce cell migration and ECM invasion. Increased invasiveness and migration correlates with enhanced expression and activation of MMP-2. Our study provides evidence showing that OPN increases gelatinolytic activity by inducing MT1-MMP expression via activation of the NF-kappaB pathway. Suppression of MMP-2 by ASMMP-2 reduces the OPN-induced cell migration and ECM invasion. Curcumin blocks OPN-induced MT1-MMP expression and pro-MMP-2 activation. Curcumin, a known anti-inflammatory and anticarcinogenic compound, suppresses OPN-induced cell migration, invasion and induces apoptotic morphology in OPN-treated cells. The mechanism by which curcumin suppresses the OPN-induced effects has also been delineated. Curcumin inhibits MT1-MMP gene expression by blocking signals leading to IKK activation. This in turn inhibits IkappaBalpha phosphorylation and NF-kappaB activation. PMID- 15750785 TI - The X-lectins: a new family with homology to the Xenopus laevis oocyte lectin XL 35. AB - The Xenopus laevis oocyte cortical granule lectin (XL35) has been studied in fertilization and embryonic development. Several nucleic acid sequences that predict proteins homologous to XL35 have since been reported in frog, human, mouse, lamprey, trout, ascidian worm. These proteins also showed high degrees of amino acid sequence homology to a common fibrinogen-like motif that may involve carbohydrate binding. Although their biological functions and carbohydrate binding specificities have not been studied in detail, this new family of lectins has common characteristics. Several independent studies on this new family of lectins strongly suggest that the lectins are expressed and stored in specialized vesicles that may be released upon the infection by pathogens. In addition, some family members have been shown to bind to oligosaccharides from bacterial pathogens. Therefore, this family of lectins likely participates in pathogen surveillance as part of the innate immune system. We propose the name X-lectin family for these homologs of XL35. PMID- 15750786 TI - Cell surface sialylation and ecto-sialyltransferase activity of human CD34 progenitors from peripheral blood and bone marrow. AB - Surface expressed negatively charged sialoglycans contribute to the regulation of adhesive cellular interactions and are thus involved in the growth and differentiation of hematopoietic progenitor cells. In particular, the cell surface sialylation state may govern the liberation of CD34+ hematopoietic precursors from bone marrow stroma cells and extracellular matrix. In order to assess the overall surface sialylation of live human CD34+ hematopoietic precursor cells, we applied a previously described flow cytometric enzyme assay. Cells with and without sialidase pretreatment were incubated in the presence of fluorescent CMP-sialic acid and exogenous ST6GalI. Thus sialylation of surface expressed lactosamine residues was analysed. We demonstrated that surface lactosamines of CD34+ precursors derived from bone marrow and peripheral blood are over 95% sialylated, predominantly in alpha2-6 linkage. These results are in accordance with flow cytometric analysis of surface lectin staining. Sialic acid specific lectins MAA and SNA were strongly bound whereas SBA, VVA, and PNA became reactive only after sialidase pretreatment. CD34+ leukemia cell lines TF1 and KG1a also showed a high degree of surface sialylation, whereas cell line KG1 expressed to the largest extent free lactosamines. In these cell lines, alpha2-6 and alpha2-3 sialylated residues were present in equal amounts. In a variation of the flow cytometric enzyme assay, live cells were incubated without exogenous STGal I to measure the activity of endogenous ecto-sialyltransferase. Ecto sialyltransferase activity was observed in all CD34+ cells which was able to resialylate major surface glycoproteins such as HLA Class I, CD45, CD43, and CD34. The ecto-sialyltransferase may serve to maintain or increase surface sialylation rapidly without de novo synthesis. PMID- 15750787 TI - Interactions between gangliosides and proteins in the exoplasmic leaflet of neuronal plasma membranes: a study performed with a tritium-labeled GM1 derivative containing a photoactivable group linked to the oligosaccharide chain. AB - Interactions between gangliosides and proteins at the exoplasmic surface of the sphingolipid-enriched membrane domains can be studied by ganglioside photolabeling combined with cell surface biotin labeling. In the present paper, we report on the results obtained using a novel radioactive photoactivable derivative of GM1 ganglioside, carrying the photoactivable nitrophenylazide group at the external galactose. After cell photolabeling with the radioactive photoactivable derivative of GM1 and cell surface biotin labeling, sphingolipid enriched domains were prepared from rat cerebellar neurons differentiated in culture and further purified by immunoprecipitation with streptavidin-coupled beads. Among proteins belonging to the sphingolipid-enriched domains that were biotin labeled, thus bearing an exoplasmic domain, a few were also cross-linked by the radioactive photoactivable ganglioside. In particular, two protein bands showing apparent molecular mass of 135 and 35 kDa were intensely photolabeled. The 135 kDa protein was immunologically identified as the GPI-anchored neural cell adhesion molecule TAG-1. These data suggest that hydrophilic interaction between the exoplasmic domains of the protein and the ganglioside sialooligosaccharide chain could exist. PMID- 15750788 TI - Multimeric glycotherapeutics: new paradigm. AB - The general principle of anti-adhesion therapy is the inhibition of microorganism adhesion to the host cell with the help of a soluble receptor analog. Despite an evident attractiveness of the concept and its long existence, the therapeutics of the 'post-antibiotic era' have not yet appeared. This can be explained by the contradictoriness of requirements for anti-adhesion drugs: to be efficient a drug must be multivalent, i.e. large molecule, but to obtain FDA approval it should be a small molecule. A way to overcome this contradiction is self-assembly of glycopeptides. The carbohydrate part of glycopeptide is responsible for binding with the lectin of microorganisms, whereas a simple peptide part is responsible for an association to the so-called tectomers. Depending on the structure, tectomers are formed either spontaneously or upon promotion of a microorganism. In particular, sialopeptide, which is capable of converting to a tectomer only in the presence of the influenza virus, has been obtained. Thus, the new strategy of anti-adhesion therapy can be formulated as follows: (1) identification of oligosaccharide-receptor for a particular virus (bacteria); (2) optimization of the peptide part; (3) conventional trials. The expected advantages of this strategy are the following: (i) no polymer; (ii) a virion completely covered with a tectomer, i.e. blocking is both complete and irreversible; (iii) rapid and rational lead identification and optimization; (iv) minimum side effects; (v) potential for microorganism resistance to natural receptor is lower than in the case of mimetics. PMID- 15750789 TI - Low expression of lipid-linked oligosaccharide due to a functionally altered Dol P-Man synthase reduces protein glycosylation in cAMP-dependent protein kinase deficient Chinese hamster ovary cells. AB - Chinese hamster ovary cells express a wide variety of glycoproteins with Mr ranging from 15,000 to 200,000 dalton and higher. Glycosylation of these proteins was much less in cAMP-dependent protein kinase (PKA)-deficient mutants which expressed either (i) a defective C-subunit with altered substrate specificity and having no detectable type II kinase (mutant 10215); or (ii) an altered RI subunit and having no detectable type II kinase (mutant 10248); or (iii) exhibited the lowest level of total kinase with no detectable type I kinase but having a small amount of type II kinase (mutant 10260). Addition of 8Br-cAMP enhanced protein glycosylation index in wild type cells 10001 by 120% but only 7 to 23% in the mutant cells. The rate of lipid-linked oligosaccharide (LLO) biosynthesis was linear for 1 h in all cell types, but the total amount of LLO expressed was much less in PKA-deficient mutants. Pulse-chase experiments indicated that the t1/2 for LLO turnover was also twice as high in PKA-deficient cells as in the wild type. Size exclusion chromatography of the mild-acid released oligosaccharide confirmed that both wild type and the mutant cells synthesized Glc3Man9GlcNAc2-PP Dol as the most predominating species with no accumulation of Man5GlcNAc2-PP-Dol in the mutants. Kinetic studies exhibited a reduced mannosylphosphodolichol synthase (DPMS) activity in mutant cells with a Km for GDP-mannose 160 to 400% higher than that of the wild type. In addition, the kcat for DPMS was also reduced 2 to 4-fold in these mutant cells. Exogenously added Dol-P failed to rescue the kcat for DPMS in CHO cell mutants; however, in vitro protein phosphorylation with a cAMP-dependent protein kinase restored their kinetic activity to the level of the wild type. PMID- 15750790 TI - Nano-biosensor development for bacterial detection during human kidney infection: use of glycoconjugate-specific antibody-bound gold NanoWire arrays (GNWA). AB - Infectious disease, commonly caused by bacterial pathogens, is now the world's leading cause of premature death and third overall cause behind cardiovascular disease and cancer. Urinary Tract Infection (UTI), caused by E. coli bacteria, is a very common bacterial infection, a majority in women (85%) and may result in severe kidney failure if not detected quickly. Among hundreds of strains the bacteria, E. coli 0157:H7, is emerging as the most aggressive one because of its capability to produce a toxin causing hemolytic uremic syndrome (HUS) resulting in death, especially in children. In the present study, a project has been undertaken for developing a rapid method for UTI detection in very low bacteria concentration, applying current knowledge of nano-technology. Experiments have been designed for the development of biosensors using nano-fabricated structures coated with elements such as gold that have affinity for biomolecules. A biosensor is a device in which a biological sensing element is either intimately connected to or integrated within a transducer. The basic principle for the detection procedure of the infection is partly based on the enzyme-linked immunosorbent assay system. Anti-E. coli antibody-bound Gold Nanowire Arrays (GNWA) prepared on anodized porous alumina template is used for the primary step followed by binding of the bacteria containing specimen. An alkaline phosphatase conjugated second antibody is then added to the system and the resultant binding determined by both electrochemical and optical measurements. Various kinds of GNWA templates were used in order to determine the one with the best affinity for antibody binding. In addition, an efficient method for enhanced antibody binding has been developed with the covalent immobilization of an organic linker Dithiobissuccinimidylundecanoate (DSU) on the GNWA surface. Studies have also been conducted to optimize the antibody-binding conditions to the linker-attached GNWA surfaces for their ability to detect bacteria in clinical concentrations. PMID- 15750792 TI - Galectins in teleost fish: Zebrafish (Danio rerio) as a model species to address their biological roles in development and innate immunity. AB - Cell surface glycans, such as glycocoproteins and glycolipids, encode information that modulates interactions between cells, or between cells and the extracellular matrix, by specifically regulating the binding to cell surface-associated or soluble carbohydrate-binding receptors, such as lectins. Rapid modifications of exposed carbohydrate moieties by glycosidases and glycosyltransferases, and the equally dynamic patterns of expression of their receptors during early development, suggest that both play important roles during embryogenesis. Among a variety of biological roles, galectins have been proposed to mediate developmental processes, such as embryo implantation and myogenesis. However, the high functional "redundancy" of the galectin repertoire in mammals has hindered the rigorous characterization of their specific roles by gene knockout approaches in murine models. In recent years, the use of teleost fish as alternative models for addressing developmental questions in mammals has expanded dramatically, and we propose their use for the elucidation of biological roles of galectins in embryogenesis and innate immunity. All three major galectin types, proto, chimera, and tandem-repeat, are present in teleost fish, and phylogenetic topologies confirm the expected clustering with their mammalian orthologues. As a model organism, the zebrafish (Danio rerio) may help to overcome limitations imposed by the murine models because it offers substantial advantages: external fertilization, transparent embryos that develop rapidly in vitro, a diverse toolbox of established methods to manipulate early gene expression, a growing collection of mutations that affect early embryonic development, availability of cell lines, and most importantly, an apparently less diversified galectin repertoire. PMID- 15750791 TI - Roles for N-glycosylation in the dynamics of Edg-1/S1P1 in sphingosine 1 phosphate-stimulated cells. AB - Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid mediator released from activated platelets. To date, 5 seven-transmembrane-spanning receptors, Edg 1/S1P1, Edg-3/S1P3, Edg-5/S1P2, Edg-6/S1P4 and Edg-8/S1P5, have been identified as specific Sph-1-P receptors. Our recent novel studies established that Edg 1/S1P1 is glycosylated in its N-terminal extracellular portion and further identified the specific glycosylation site as asparagine 30. We also demonstrated that the structure of the N-terminal ectodomain of Edg-1/S1P1 affects both its transport to the cell surface and the N-glycosylation process. These studies revealed a possible regulatory role for the N-glycan on Edg-1/S1P1 in the dynamics of the receptor, such as its lateral and internal movements within the membrane, in ligand-stimulated mammalian cells. PMID- 15750796 TI - Ischemic colitis following colonoscopy in a systemic lupus erythematosus patient: report of a case. AB - BACKGROUND: Ischemic colitis is an uncommon complication in patients with systemic lupus erythematosus but may be precipitated by colonoscopy. CASE REPORT: A 43-year-old female with systemic lupus erythematosus under treatment with immunosuppressive drugs and prednisone was submitted to colonoscopy because of a change in bowel habit. Apart from the presence of a small metaplastic polyp, colonoscopy showed only a few erythematosus areas in the sigma and left colon. Four hours after colonoscopy, the patient developed lower colic abdominal pain and mucous diarrhea followed by rectal bleeding from ischemic colitis. The patient was successfully treated with fluids, spasmolytic drugs, sodic heparin, antibiotics and enteral feeding. CONCLUSIONS: Awareness of the risk of this potential complication, secondary to colonoscopy, in patients with connective tissue disorders may lead to a prompt diagnosis and effective treatment, with a successful outcome. PMID- 15750797 TI - Anti-TNF-alpha (infliximab) used as induction treatment in case of active proctitis in a multistep strategy followed by definitive surgery of complex anal fistulas in Crohn's disease: a preliminary report. AB - PURPOSE: This study was designed to assess the healing rate of complex perianal fistulas in Crohn's disease after a multistep strategy, including induction treatment with Infliximab in case of active proctitis, followed by definitive surgery. METHODS: From 2000 to 2003, all consecutive patients with complex fistulas and Crohn's disease underwent pretreatment with noncutting setons and, in case of severe recurrent fistulas or abscesses, a diverting stoma. Infliximab was added in cases of active proctitis. After definitive surgical treatment, patients were examined. RESULTS: Seventeen patients were included (median age, 34 (range, 22-58) years). Seven patients were treated by surgery only, and in ten patients Infliximab was added. After a median follow-up of 19 (range, 8-40) months, fistula healing was observed in 17 patients (100 percent). One patient of the Infliximab group developed a recurrent fistula (10 percent) after 24 months, and in one patient (10 percent) soiling occurred. Two patients of the surgical group developed a recurrent fistula (29 percent) and soiling occurred in two patients (29 percent). CONCLUSIONS: A multistep strategy followed by definitive surgery for the treatment of complex perianal fistulas in patients with Crohn's disease is a promising treatment modality. The preliminary results of this study suggest that Infliximab treatment has a beneficial additive effect in the multistep treatment followed by definitive surgery of complex anal fistulas and active proctitis in Crohn's disease. PMID- 15750798 TI - An international survey of training in adult intensive care medicine. AB - OBJECTIVE: The aim of this international survey of training in adult intensive care medicine (ICM) was to characterise current structures, processes, and outcomes to determine the potential for convergence to a common competency-based training programme across national borders. This survey is the first phase of a 3 year project which will use consensus methods to build an international competency-based training programme in ICM in Europe (CoBaTrICE). METHODOLOGY: A survey by questionnaire, e-mail, and direct discussion was undertaken with national ICM representatives from seven geographical regions. RESULTS: Responses were obtained from 41 countries (countries which share common training programmes were grouped together; n=38). Fifty-four different training programmes were identified, 37 within the European region; three (6%) were competency-based. Twenty (53%) permitted multidisciplinary access to a common training programme; in nine (24%) training was only available within anaesthesia. The minimum duration of ICM training required for recognition as a specialist varied from 3 months to 72 months (mode 24 months). The content of most (75%) ICM programmes was standardised nationally. Work-based assessment of competence was formally documented in nineteen (50%) countries. An exam was mandatory in twenty-nine (76%). CONCLUSION: There are considerable variations in the structures and processes of ICM training worldwide. However, as competency-based training is an outcome strategy rather than a didactic process, these differences should not impede the development of a common international competency-based training programme in ICM. PMID- 15750799 TI - Withholding and withdrawal of life-sustaining treatment in a Lebanese intensive care unit: a prospective observational study. AB - OBJECTIVE: To evaluate the implementation and process of withholding and withdrawing life-sustaining treatment in an intensive care unit. DESIGN AND SETTING: Prospective observational study in the medical intensive care unit of a university hospital in Lebanon. PATIENTS: Forty-five consecutive adult patients admitted to the ICU for a 1-year period and for whom a decision to withholding and withdrawal of life-sustaining treatment was made. MEASUREMENTS AND RESULTS: Patients were followed up until their death. Data regarding all aspects of the implementation and the process of withholding and withdrawal of life-sustaining treatment were recorded by a senior staff nurse. Withholding and withdrawing life sustaining treatment was applied to 9.6% of all admitted patients to ICU. Therapies were withheld in 38% and were withdrawn in 7% of patients who died. Futility of care and poor quality of life were the two most important factors supporting these decisions. The nursing staff was not involved in 26% of the decisions to limit care. Families were not implicated in 21% of the cases. Decisions were not notified in the patients' medical record in 23% of the cases. Sixty-three percent of patients did not have a sedative or an analgesic to treat discomfort during end-of-life care. CONCLUSIONS: Life-sustaining treatment were frequently withheld or withdrawn from adult patients in the Lebanese ICU. Cultural differences and the lack of guidelines and official statements could explain the ethical limitations of the decision-making process recorded in this study. PMID- 15750800 TI - ICU physicians should abandon the use of etomidate! PMID- 15750801 TI - Coronary artery bypass graft surgery for persistent weaning failure. PMID- 15750802 TI - Metabolic monitoring by bacterial mRNAs. AB - There is growing appreciation for diversity in the strategies that bacteria utilize in regulating gene expression. Bacteria must be able to respond in different ways to different stresses and thus require unique regulatory solutions for the physiological challenges they encounter. Recent data indicate that bacteria commonly employ a variety of posttranscriptional regulatory mechanisms to coordinate expression of their genes. In many instances, RNA structures embedded at the 5' ends of mRNAs are utilized to sense particular metabolic cues and regulate the encoded genes. These RNA elements are likely to range in structural sophistication, from short sequences recognized by RNA-binding proteins to complex shapes that fold into high-affinity receptors for small organic molecules. Enough examples of RNA-mediated genetic strategies have been found that it is becoming useful to view this overall mode of regulatory control at a genomic level. Eventually, a complete picture of bacterial gene regulation within a single bacterium, from control at transcription initiation to control of mRNA stability, will emerge. But for now, this article seeks to provide a brief overview of the known categories of RNA-mediated genetic mechanisms within the bacterium Bacillus subtilis, with the expectation that it is representative of bacteria as a whole. PMID- 15750803 TI - Transepidermal water loss and skin capacitance alterations among workers in an ultra-low humidity environment. AB - No studies have been performed evaluating skin barrier alterations in humans exposed to ultra-low humidity (ULH) in spite of several lines of evidence from animal experiments suggesting that the skin barrier is altered on exposure to ULH. The objectives of this study were to assess barrier function changes in workers occupationally exposed to ULH (relative humidity 1.5%), and to evaluate whether the exposure duration shows a dose-response relationship with transepidermal water loss (TEWL) and skin capacitance. A total of 49 male workers exposed to ULH for 12 h per working day were classified into five subgroups based on their ULH exposure duration (<0.5 months, 0.5-1.0 month, 1.1-10.0 months, 10.1 20.0 months, and >20.0 months). A group of 12 age-matched male laboratory workers from a normal humidity environment were recruited as a control group. TEWL and skin capacitance were measured to evaluate their skin barrier function. TEWL measurements showed a significant decline (8.3+/-0.4 vs 10.0+/-0.4 g m(-2) h(-1), P < 0.05) but no differences were found in skin capacitance (39.7+/-1.3 a.u. vs 45.0+/-2.4 a.u., P = 0.68) between the whole ULH exposure group and the control group. Maximum decreases in TEWL and skin capacitance were seen in the subgroups exposed for <0.5 months and 0.5-1.0 month, respectively. Almost completely natural recovery occurred in skin capacitance after 20 months ULH exposure, in contrast to less than 90% recovery in TEWL. Three stages were defined according to the pattern of alterations in TEWL and skin capacitance in relation to ULH exposure duration. A positive association between TEWL and skin capacitance occurred in the control group and stage I but a negative correlation in stage II. No correlation was found in stage III. Our study demonstrated that workers exposed to a ULH environment could exhibit skin barrier alterations. Both TEWL and skin capacitance decreased within 2 weeks of ULH exposure. The maximum alterations in TEWL and skin capacitance occurred during 0.5-1.0 months and 2 weeks, respectively. TEWL recovered partially and more slowly than skin capacitance which recovered earlier and almost completely. PMID- 15750805 TI - Egocentric search for disappearing objects in domestic dogs: evidence for a geometric hypothesis of direction. AB - In several species, the ability to locate a disappearing object is an adaptive component of predatory and social behaviour. In domestic dogs, spatial memory for hidden objects is primarily based on an egocentric frame of reference. We investigated the geometric components of egocentric spatial information used by domestic dogs to locate an object they saw move and disappear. In experiment 1, the distance and the direction between the position of the animal and the hiding location were put in conflict. Results showed that the dogs primarily used the directional information between their own spatial coordinates and the target position. In experiment 2, the accuracy of the dogs in finding a hidden object by using directional information was estimated by manipulating the angular deviation between adjacent hiding locations and the position of the animal. Four angular deviations were tested: 5, 7.5, 10 and 15 degrees . Results showed that the performance of the dogs decreased as a function of the angular deviations but it clearly remained well above chance, revealing that the representation of the dogs for direction is precise. In the discussion, we examine how and why domestic dogs determine the direction in which they saw an object disappear. PMID- 15750804 TI - Arterial intima-media thickening and endothelial dysfunction in obese Chinese children. AB - We studied ultrasonographic signs of early atherosclerosis in relation to established risk factors of cardiovascular disease in 43 obese school-age children compared to 28 non-obese controls. The groups did not show significant differences in age, gender ratio and body height. Mean carotid intima-media thickness (IMT), carotid haemodynamic parameters, flow-mediated dilatation of the brachial artery (FMD), and biochemical markers of dyslipidaemia were measured and correlated. IMT was significantly increased (0.62 mm vs. 0.46 mm, P <0.001) and FMD was markedly reduced (10.9% vs. 18.8%, P <0.001) in the obesity group. Differences in IMT and FMD persisted after adjustment for Body Mass Index (BMI) and blood pressure on ANCOVA. BMI was highly associated with increased IMT ( r =0.58, P <0.01) and reduced FMD ( r =-0.42, P <0.01), as were blood levels of total cholesterol, LDL cholesterol and Apo B. CONCLUSION: Childhood obesity seems to contribute to the development and progression of early atherosclerosis, particularly in combination with hypertension and dyslipidaemia. In order to prevent coronary atherosclerosis and other cardiovascular complications, it is vital to control obesity starting from childhood. Ultrasonography of the arterial wall may be used in a clinical setting to identify high-risk patients among severely obese children. PMID- 15750806 TI - Polysaccharide-producing bacteria isolated from paper machine slime deposits. AB - Development of novel enzymatic methods for slime deposit control in paper mills requires knowledge of polysaccharide-producing organisms and the polysaccharide structures present in deposits. In this work, 27 polysaccharide-producing bacteria were isolated from slime samples collected from different parts of a paper machine. Most of the isolates produced polysaccharides in liquid culture and nine of them were selected for production of polysaccharides for characterisation. The selected isolates belonged to seven different genera: Bacillus, Brevundimonas, Cytophaga, Enterobacter, Klebsiella, Paenibacillus and Starkeya. Using ribotyping, partial 16S rDNA sequencing, physiological tests and fatty acid analysis, four of the nine isolates: Bacillus cereus, Brevundimonas vesicularis, K. pneumoniae and P. stellifer were identified to the species level. Production of polysaccharides by the selected isolates varied between 0.07 and 1.20 g L(-1), the highest amount being produced by B. vesicularis. The polysaccharides were heteropolysaccharides with varying proportions of galactose, glucose mannose, rhamnose fucose and uronic acids. PMID- 15750807 TI - Light effects on cell development and secondary metabolism in Monascus. AB - In nature, light is one of most crucial environmental signals for developmental and physiological processes in various organisms, including filamentous fungi. We have found that both red light and blue light affect development in Monascus, influencing the processes of mycelium and spore formation, and the production of secondary metabolites such as gamma-aminobutyric acid, red pigments, monacolin K and citrinin. Additionally, we observed that the wavelength of light affects these developmental and physiological processes in different ways. These findings suggest that Monascus possesses a system for differential light response and regulation. PMID- 15750808 TI - Bioavailability of phenanthrene in the presence of birnessite-mediated catechol polymers. AB - Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants and contribute to the pollution of aquatic and terrestrial environments. In soil, their fate may be affected by interactions with the soil biological community and soil colloids. This study was conducted to investigate the fate of phenanthrene (Phe), selected as a representative PAH, in simplified model systems, which simulate processes naturally occurring in soil. Phe was interacted with catechol (Cat), an orthodiphenol, and common intermediate in the microbial degradation of PAHs, and birnessite (Bir), an abiotic strong oxidative catalyst abundant in soil. Two experimental conditions were investigated: Cat (5 mM)+Bir (1 mg ml( 1))+Phe (0.05 mg ml(-1)) mixed at the same time and incubated for 24 h at 25 degrees C (Cat-Bir-Phe) and Cat+Bir incubated for 24 h at 25 degrees C before Phe addition and then incubated for a further 24 h (Cat-Bir+Phe). After incubation, the systems were analysed for residual Cat and Phe, supplied with a selected Phe degrading mixed bacterial culture, and then the microbial degradation of Phe and the growth of cells were monitored. Complex phenomena simultaneously occurred. Cat was completely removed after a 24-h incubation with Bir, and no interference by Phe in the Bir-mediated transformation of Cat was observed. Elemental analysis and UV-Vis and Fourier transfer infrared spectra showed that Cat transformation by Bir produced soluble and insoluble polymeric aggregates involving Phe. The hydrocarbon also interacted with the surfaces of Bir either previously coated (Cat-Bir+Phe sample) or not by Cat polymers. When a Phe-degrading bacterial culture was added to the systems after Bir-mediated Cat polymerisation, a different behaviour was observed in terms of Phe consumption and bacterial growth, thus suggesting differentiated availability of Phe to the microbial cells. The hydrocarbon was completely transformed in the presence of Bir and/or Bir covered by Cat polymers. By contrast a reduced degradation was measured when the Phe was involved in the polymerisation of Cat and entrapped in the Cat polymers (Cat-Bir-Phe). Although Cat showed a toxic, lethal effect on the bacterial cells, microbial growth was observed in the presence of Cat and Cat polymers, as the only C source. The mechanism leading to the different availability of Phe in the presence of Cat and Bir is still not clear. Further investigations are requested to provide more insight into such a complex phenomenon. PMID- 15750809 TI - Sudden cardiac death in a child affected by Prader-Willi syndrome. AB - A case of sudden cardiac death in a 3-year-old young male affected by Prader Willi syndrome, clinically diagnosed and confirmed by means of DNA methylation, is presented. The infant suddenly collapsed at home and was taken apparently unconsciousness by his mother to the emergency clinic where he was pronounced dead. A complete postmortem examination was performed and the histological findings led to the definition of cardiac death with a typical picture of contraction band necrosis. Pulmonary hypoxic alterations are frequently reported as the primary cause of death in PWS cases. In this fatal case according to the macroscopic and microscopic findings, the cause of death was most likely cardiac and possibly related to contraction band necrosis linked with ventricular fibrillation and sudden death. PMID- 15750811 TI - Harmonic versus conventional ultrasound imaging of the urinary tract in children. AB - BACKGROUND: Harmonic imaging (HI) is a relatively new US method. Its usefulness in children has yet to be determined. OBJECTIVE: To evaluate the value of HI in urinary tract imaging of infants and children. MATERIALS AND METHODS: The bladder and kidneys of 29 unselected patients, aged 2 months to 12.8 years (mean, 6.8 years), were examined from ventral and dorsal approaches using conventional (= fundamental) imaging (FI) (6.5 and 3.4 MHz) and HI (3.3 MHz). In addition to global image quality, visualization of bladder wall, retrovesical space, renal contrast to liver/spleen, the pelvicalyceal systems, and the difference between cortex and medulla were assessed. Parameters were rated by two independent examiners and statistical analysis was performed. RESULTS: There were significant differences between the three imaging settings for all parameters studied (P<0.003). With the dorsal approach, HI was superior to FI for all parameters analysed (P<0.05). Using the ventral approach, a significant improvement of imaging with HI was found for the bladder and the renal pelvis (P<0.02). CONCLUSIONS: HI is a useful additional tool for imaging the urinary tract in children of all ages. The method particularly improves visualization of the bladder and the kidney from a dorsal approach. PMID- 15750812 TI - Merosin-deficient congenital muscular dystrophy (CMD): a study of 25 Brazilian patients using MRI. AB - BACKGROUND: Merosin-deficient congenital muscular dystrophy (CMD) is characterized clinically by hypotonia and muscular weakness and, on imaging studies, by white matter (WM) abnormality. OBJECTIVE: To evaluate MRI findings in Brazilian patients with merosin-deficient CMD. MATERIALS AND METHODS: Twenty-five patients were evaluated using MRI. Three patients presented with partial merosin deficiency and 22 with total merosin deficiency. Follow-up examinations were done in 7 cases. T1- and T2-weighted images were performed in all examinations, and fluid-attenuated inversion recovery (FLAIR) was performed in 15. Enhanced images were done in 11 cases. The WM involvement was classified according to location and severity. RESULTS: From 1991 to 2004, 32 MRI examinations were performed. Severe involvement was found in 23 patients in the frontal and temporal lobes, in 18 patients in the parietal lobes, and in 7 patients in the occipital lobes. The brain stem (n=5), cerebellum (n=6), internal capsules (n=1), and external capsules (n=5) were also affected. One patient had occipital pachygyria, and one had cerebellar vermian hypoplasia. No gadolinium enhancement was noted. Follow-up MRI showed no interval change (n=4), progression (n=1), or improvement of the findings (n=2). CONCLUSION: This series of patients demonstrated that there was no correlation between the extent of WM abnormality on MRI and the clinical status and degree of merosin deficiency (partial or total). Bilateral WM involvement was seen to be more prominent in the parietal, frontal, and temporal regions of the brain. The brain stem and internal and external capsules were less affected. Cerebellar WM involvement is rare. Changes on follow-up imaging studies did not correlate with the clinical status of the patient. PMID- 15750813 TI - Statistical approaches to language acquisition and the self-organizing consciousness: a reversal of perspective. AB - Recent years have seen the upsurge of a new approach to language that moves away from the rule-based conventional framework. In this approach, mostly supported by the success of connectionist models, children learn language by exploiting the distributional properties of the input. It is argued in this paper that, in the same way as conforming to rules does not imply the existence of mental rules, conforming to statistical regularities does not imply that statistical computations are performed mentally. Sensitivity to statistical regularities can alternatively be conceived of as a by-product of the recurrent interplay between the properties of the current conscious content and the properties of the linguistic and extralinguistic environment. The validity of including the content of conscious experiences in an otherwise standard dynamical approach rooted in the notion of self-organization is discussed. PMID- 15750814 TI - Empirical research on the generation and functional role of consciousness. PMID- 15750815 TI - Influence of intracoronary attenuation on coronary plaque measurements using multislice computed tomography: observations in an ex vivo model of coronary computed tomography angiography. AB - Assessment of attenuation (measured in Hounsfield units, HU) of human coronary plaques was performed using multislice computed tomography (MSCT) in an ex vivo model. In three ex vivo specimens of left coronary arteries in oil, MSCT was performed after intracoronary injection of four solutions of contrast material (400 mgI/ml iomeprol). The four solutions were diluted as follows: 1/infinity, 1/200, 1/80, and 1/20. All scans were performed with the following parameters: slices/collimation 16/0.75 mm, rotation time 375 ms. Each specimen was scored for the presence of atherosclerotic plaques. In each plaque the attenuation was measured in four regions of interest for lumen, plaque (non-calcified thickening of the vessel wall), calcium, and surrounding (oil surrounding the vessel). The results were compared with a one-way analysis of variance test and were correlated with Pearson's test. There were no significant differences in the attenuation of calcium and oil in the four solutions. The mean attenuation in the four solutions for lumen (35+/-10, 91+/-7, 246+/-18, 511+/-89 HU) and plaque (22+/-22, 50+/-26, 107+/-36, 152+/-67 HU) was significantly different between each decreasing dilution (p<0.001). The mean attenuation of lumen and plaque of coronary plaques showed high correlation, while the values were significantly different (r=0.73; p<0.001). Intracoronary attenuation modifies significantly the attenuation of plaques assessed with MSCT. PMID- 15750816 TI - Decrease in excitability of LG following habituation of the crayfish escape reaction. AB - Crayfish escapes from threatening stimuli to the abdomen by tailflipping upwards and forwards. This lateral giant (LG)-mediated escape reaction habituates readily upon repetitive sensory stimulation. Using an isolated abdominal nerve cord preparation, we have analyzed the change in LG activity by applying additional sensory stimulation after different periods following habituation to characterize the retention of LG habituation. Results show that the LG mediated response habituates more quickly, but the retention time is shorter, as repetitive sensory stimulation is applied at progressively shorter inter-stimulus time intervals. The spike response of LG recovers quickly, within several minutes after habituation, but they fail to spike when an additional stimulus is applied after specific long periods following habituation. The critical period of the delay for this decrease in excitability of LG is dependent on the inter-stimulus time interval of the initial repetitive stimulus. As the inter-stimulus interval became longer, the delay needed for decrease in excitability became shorter. Furthermore, the local injection of 10(-6) mol l-1 octopamine into the neuropil just following habituation promotes the achievement of decrease in excitability. No effects were observed when 10(-6) mol l-1 serotonin and tyramine were injected. These results suggested octopamine promotes decrease in excitability of LG following habituation. PMID- 15750817 TI - Long-term potentiation and olfactory memory formation in the carp (Cyprinus carpio L.) olfactory bulb. AB - Long-term potentiation of synaptic transmission is considered to be an elementary process underlying the cellular mechanism of memory formation. In the present study we aimed to examine whether or not the dendrodendritic mitral-to-granule cell synapses in the carp olfactory bulb show plastic changes after their repeated activation. It was found that: (1) the dendrodendritic mitral-to-granule cell synapses showed three types of plasticity after tetanic electrical stimulation applied to the olfactory tract-long-term potentiation (potentiation lasting >1 h), short-term potentiation (potentiation lasting <1 h) and post tetanic potentiation (potentiation lasting <10 min); (2) Long-term potentiation was generally induced when both the dendrodendritic mitral-to-granule cell synapses and centrifugal fiber-to-granule cell synapses were repeatedly and simultaneously activated; (3) long-term enhancement (>1 h) of the odor-evoked bulbar response accompanied the electrically-induced LTP, and; (4) repeated olfactory stimulation enhanced dendrodendritic mitral-to-granule cell transmission. Based on these results, it was proposed that long-term potentiation (as well as olfactory memory) occurs at the dendrodendritic mitral-to-granule cell synapses after strong and long-lasting depolarization of granule cells, which follows repeated and simultaneous synaptic activation of both the peripheral and deep dendrites (or somata). PMID- 15750818 TI - Slow adaptation in spider mechanoreceptor neurons. AB - Slow adaptation of action potential firing is a common but poorly understood property of sensory neurons. We quantified slow adaptation in a cuticular mechanoreceptor organ of the spider, Cupiennius salei, by stimulating with continuous pseudorandom mechanical displacements while recording action potentials intracellularly from the cell bodies. Firing rate declined over a period of several minutes before reaching a steady level at about half the initial rate. This slow adaptation was fitted by an exponential decay with mean time constant of 18.5 s. Recovery from slow adaptation was also fitted by an exponential process, but with a longer time constant of 167 s. The receptor potential produced by the same stimulation protocol did not change its amplitude or dynamics, showing that slow adaptation occurs during action potential encoding from the receptor potential. Experiments with chemical blockers of calcium entry or the known potassium currents failed to reduce the slow adaptation. The Na+/K+ pump blocker Ouabain decreased the time constant of slow adaptation, suggesting that ion accumulation is involved. In some experiments, a second class of small action potentials were observed, which were tentatively attributed to failed conduction from the sensory dendrite through the soma to the axon. PMID- 15750820 TI - Magnetic field exposure and arrythmic risk: evaluation in railway drivers. AB - Recent studies suggest that professional exposure to Extremely Low Frequency Electro Magnetic Field (ELF-EMF) can increase the risk of sudden cardiac death. Aim of our work was to find predictive parameters of arrhythmic risk in a population of 28 railways drivers exposed to ELF-EMF. Our findings were that the exposure did not reduce HRV and did not increase the risk of arrhythmias. PMID- 15750819 TI - Exposure to air pollution and pulmonary function in university students. AB - OBJECTIVES: Exposure to air pollution has been reported to be associated with increase in pulmonary disease. The aims of the present study were to examine the use of personal nitrogen dioxide (NO(2)) samplers as a means of measuring exposure to air pollution and to investigate the relationship between personal exposure to air pollution and pulmonary function. METHODS: We measured individual exposures to NO(2) using passive personal NO(2) samplers for 298 healthy university students. Questionnaire interview was conducted for traffic-related factors, and spirometry was performed when the samplers were returned after 1 day. RESULTS: Personal NO(2) concentrations varied, depending on the distance between residence and a main road (P=0.029). Students who used transportation for more than 1 h were exposed to higher levels of NO(2) than those using transportation for less than 1 h (P=0.032). In terms of transportation, riding in a bus or subway caused significantly higher exposure than not using them (P=0.046). NO(2) exposure was not significantly associated with forced vital capacity (FVC) or forced expiratory volume in 1 s (FEV(1)) but was associated with the ratio of FEV(1)/FVC and mid-expiratory flow between 25% and 75% of the forced vital capacity (FEF(25-75)) (P<0.05). CONCLUSIONS: This study indicates that concentrations of personal exposure to NO(2) are significantly influenced by traffic-related air pollution and are associated with decreased pulmonary function. PMID- 15750821 TI - Lead in finger bone, whole blood, plasma and urine in lead-smelter workers: extended exposure range. AB - OBJECTIVES: To assess the historical exposure and to study the relationships between lead concentrations in whole blood (B-Pb), plasma (P-Pb), urine (U-Pb), finger bone (Bone-Pb) and duration of employment in workers at a secondary lead smelter and to compare the relationships between B-Pb and P-Pb with results from previous studies of populations with a wide range of lead exposure. METHODS: In 39 lead workers (29 active, ten retired), recruited from those with the highest exposure at a German secondary lead smelter, levels of B-Pb, P-Pb and U-Pb were determined by inductively coupled plasma mass spectrometry (ICP-MS). Bone-Pb was determined by in vivo X-ray fluorescence (XRF). Results were compared with data from a previous study on 90 workers (71 active, 19 retired) with lower exposure, from a Swedish secondary lead smelter, as well as with previously collected data from 42 active Russian lead workers and 34 Ecuadorian lead-exposed subjects. RESULTS: The median values in the active/retired German lead workers were: age 44/59 years, duration of employment 20/38 years, Bone-Pb 71/150 microg/g, B-Pb 500/330 microg/l, P-Pb 2.7/1.1 microg/l, and U-Pb 25/13 micromol/mol creatinine. Bone-Pb increased with duration of employment by 4.2 microg/g per year and 1.6 microg/g per year in German and Swedish workers, respectively. The median Bone-Pb was three times higher in both active and retired German workers than in Swedish smelter workers with essentially the same age distribution and duration of employment. The linear regression equation between B-Pb and log P-Pb in the combined group of Ecuadorian, German, Russian and Swedish lead-exposed subjects (n=176) was B-Pb=545 x log[P-Pb] + 258 (r(s)=0.94; P<0.001). CONCLUSIONS: The high Bone-Pb values recorded for the German smelters implied a historical lead exposure of considerable magnitude. The long-term high lead exposure also showed up in the B-Pb levels for both active and retired workers, leading to the implementation of necessary industrial safety measures in order to respond to biological threshold limits. The suggested equation describing the relationship between B-Pb and P-Pb in the combined group of subjects with a wide range of lead exposure can be useful in future cross-sectional and longitudinal studies of lead exposed populations, relating, e.g., lead exposure to adverse health outcomes. PMID- 15750822 TI - Function associated transforming growth factor-beta gene polymorphism in chronic beryllium disease. AB - Chronic beryllium disease (CBD) is a rare occupational, granulomatous lung disease clinically resembling sarcoidosis. The immune response to beryllium is thought to depend on genetic susceptibility. Although a glutamic acid in position 69 of the human leukocyte antigen-DP beta chain (HLA-DPB1-Glu69) is associated with the development of CBD, it cannot fully explain susceptibility. It is likely that additionally other genes are involved in regulating the immune and inflammatory response in the pathogenesis of this disease. Functional gene polymorphisms (PMs) of the tumor necrosis factor (TNF)A and transforming growth factor (TGF) beta(1) genes are suspected to modify the course of granulomatous disorders. We analyzed the TGF-beta(1) (codon 25) PM in 59 patients with CBD and 164 matched healthy controls, from two groups of European/Israeli and United States origin. Additionally, patients were genotyped for HLA class II gene variants and the TNFA (-308) PM. The most significant results were found for the TGF-beta(1) (codon 25) PM with a shift towards the low producing non-GG genotypes in the subgroup of European and Israeli patients with CBD (62.50% vs. 13.82% in healthy controls; P<0.001). This phenomenon was not observed in the group from the United States. Moreover, TGF-beta(1) (codon 25) PM genotype frequencies from United States CBD patients differed significantly from those of European and Israeli patients. In contrast, increased frequencies for the high producing TNFA2 allele were found only in the patients from the United States (28.20% vs. 8.96% in healthy controls; P<0.005) but not in the group of Europe and Israel. In conclusion, the increase in TGF-beta(1) (codon 25) PM genotype frequency associated with a low TGF-beta release suggests that immunoregulatory cytokines such as TGF-beta are involved in the pathogenesis of CBD. Moreover, based on the interaction of gene PMs associated with the control of the immune response, such as TNF-alpha and TGF-beta(1), with a specific immune response gene such as HLA DPB1-Glu69 or other HLA-class II PMs driving the immune response to Be, the present data suggest that a combination of different genetic backgrounds determine susceptibility for the same immunopathological reaction and disease. PMID- 15750824 TI - Nasal Staphylococcus aureus carriage is not increased in Behcet's disease. PMID- 15750823 TI - Erythromelalgia as a presenting manifestation in a patient with essential thrombocythemia complicating renovascular hypertension due to unilateral renal artery stenosis. AB - Erythromelalgia is a kind of cutaneous manifestation, which appears as a thrombotic complication in patients with myeloproliferative disorders such as essential thrombocythemia and polycythemia vera. It is characterized by red, congested distal extremities and a painful burning sensation, and is usually confined to the feet and one or more toes or fingers. A 28-year-old woman visited our hospital due to severe pain in the left thumb, index fingers and right toes. Her right toes and left thumb were erythematous, congested, and warm. She had a high blood pressure level of 190/100 mmHg, and laboratory evaluation revealed marked thrombocytosis. Bone-marrow findings were compatible with essential thrombocythemia. Renal angiography showed obvious stenosis in unilateral right renal artery. Her erythromelalgia immediately disappeared following interventional therapy along with aspirin. A careful history and appropriate evaluation of underlying diseases are important, because erythromelalgia as a microscopic thrombotic complication may be accompanied by vascular stenosis and all the resulting manifestations. PMID- 15750825 TI - DNA markers associated with low Fusarium head blight incidence and narrow flower opening in wheat. AB - Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum, is an important fungal disease in many wheat-growing areas of the world. The objectives of this study were to determine the relationship between width and duration of flower opening and incidence of FHB in wheat, and to identify DNA markers associated with narrow flower opening and low FHB incidence. It was hypothesized that wheat lines whose flowers open briefly and narrowly have a reduced risk of infection. To test the hypothesis, we crossed wheat cultivars Patterson and Goldfield to generate a population of 100 random F(2)-derived recombinant inbred lines (RILs). Florets of Patterson open wide; florets of Goldfield tend to stay closed. The population of RILs was characterized for FHB incidence and flower opening width (FOW) and duration in the F(7:9) and F(7:10) generations. Of the 305 simple sequence repeat primer pairs screened on the parents, 79 amplified polymorphic DNA bands. Pooled DNA from each of the two bulks was tested with these 79 SSR primer pairs. Four markers were found to have significant marker trait association with low FHB incidence and narrow flower opening. The major QTL effect associated with narrow flower opening and low FHB incidence was found between the map interval Xbarc200-Xgwm210, explaining 29% of the phenotypic variation for FHB incidence averaged over six replicated tests in Indiana in 2002 and 2003. This adds credence to the hypothesis that narrow flower opening is responsible for low FHB incidence in this population. Breeding wheat lines for both morphological avoidance, such as narrow flower opening, and physiological resistance to FHB may be valuable in future breeding research to reduce crop production and grain quality losses in wheat due to FHB. PMID- 15750826 TI - Molecular marker-based genetic diversity assessment of Striga-resistant maize inbred lines. AB - Striga-resistant maize inbred lines are of interest to maize breeding programs in the savannas of Africa where the parasitic weed is endemic and causes severe yield losses in tropical maize. Assessment of the genetic diversity of such inbred lines is useful for their systematic and efficient use in a breeding program. Diversity analysis of 41 Striga-resistant maize inbred lines was conducted using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers to examine the genetic relationships among these lines and to determine the level of genetic diversity that exists within and between their source populations. The two marker systems generated 262 and 101 polymorphic fragments, respectively. Genetic similarity (GS) values among all possible pairs of inbred lines varied from 0.45 to 0.95, with a mean of 0.61+/-0.002 for AFLPs, and from 0.21 to 0.92, with a mean of 0.48+/-0.003, for SSRs. The inbred lines from each source population exhibited a broad range of GS values with the two types of markers. Both AFLPs and SSRs revealed similar levels of within population genetic variation for all source populations. Cluster and principal component analysis of GS estimates with the two markers revealed clear differentiation of the Striga-resistant inbred lines into groups according to their source populations. There was clear separation between early- and late maturing Striga-resistant inbred lines. Considering the paucity of germplasm with good levels of resistance to Striga in maize, the broad genetic diversity detected within and among source populations demonstrates the genetic potential that exists to improve maize for resistance to Striga. PMID- 15750827 TI - Practice guidelines based on clinical and economic evidence. Indispensable tools in future market oriented health care. PMID- 15750828 TI - Heterogeneity of cerebral blood flow in dementia. PMID- 15750829 TI - Sweet nuclear medicine phantoms for scintigraphic sentinel lymph node detection: a cooking recipe. PMID- 15750830 TI - Epithelial tight junction proteins as potential antibody targets for pancarcinoma therapy. AB - Recombinant monoclonal antibodies are beginning to revolutionize cancer therapy. In combination with standard chemotherapy, high response rates have been reported with antibodies of the human IgG1 isotype for treatment of non-Hodgkin's lymphoma and breast cancer. It is becoming apparent that targets for antibody-based therapies do not necessarily need to be absent from normal tissues but can be present there either in low copy numbers or with binding epitopes shielded from the therapeutic antibody. Here, we studied whether claudin proteins that form tight junctions in normal epithelia are still expressed on carcinoma cells and whether their extracellular domains can be recognized by antibodies. We show that mRNAs of claudins 1, 3, 4, and 7 are all expressed in different human carcinoma cell lines, while claudin 8 was selectively expressed in breast and pancreas cancer lines. Chicken polyclonal antibodies were raised against peptides contained within predicted extracellular domains of claudins 1, 3, and 4. Affinity-purified IgG fractions for claudins 3 and 4 were monospecific and bound to human breast and colon carcinoma lines, but not to a line of monocytic origin. Claudin 3 antibodies also homogeneously stained human renal cell carcinoma tissue and micrometastatic tumor cells as identified by cytokeratin staining in bone marrow biopsies of breast cancer patients. Fluorescence-activated cell sorting and immunocytochemistry indicated that claudin antibodies bound to the surface of tumor cells. By analogy to other tumor-associated antigens that are differentially accessible to antibodies on tumor vs normal tissue, we propose that certain claudin proteins have potential as targets for novel antibody-based therapies of carcinomas. PMID- 15750831 TI - Effective induction of antitumor immunity by immunization with plasmid DNA encoding TRP-2 plus neutralization of TGF-beta. AB - Plasmid DNA vaccine is an appealing cancer immunotherapy. However, it is a weak immunogen and immunization with plasmid DNA encoding self-antigens, such as melanoma-associated antigens, could not induce antitumor immunity because of tolerance. In this study, we investigated the feasibility of using a plasmid DNA encoding Xenopus laevis transforming growth factor-beta 5 (aTGF-beta5) as an immunogen to induce neutralizing antibodies against murine TGF-beta1 (mTGF-beta1) and thus enhance the efficacy of plasmid DNA vaccine encoding murine tyrosinase related protein 2 (mTRP-2) through neutralization of TGF-beta. The results showed that immunization with aTGF-beta5 resulted in the generation of mTGF-beta1 neutralizing antibodies, and immunization with a combination of aTGF-beta5 and mTRP-2 induced specific cytotoxic T lymphocytes (CTLs). On the contrary, immunization with mTRP-2 alone could not elicit the CTL response. Moreover, immunization of C57BL/6 wild-type mice with a combination of aTGF-beta5 and mTRP 2 induced the protective and therapeutic antitumor immunity to B16F10 melanoma, whereas the antitumor activity was abrogated in both CD4-deficient mice and CD8 deficient mice on the C57BL/6 background. Our results indicate that immunization with aTGF-beta5 is capable of breaking immune tolerance and induces mTGF-beta1 neutralizing antibodies. Neutralization of TGF-beta can enhance the efficacy of DNA vaccine encoding mTRP-2 and the induction of antitumor immunity by this immunization strategy is associated with CD4+ and CD8+ T cells. PMID- 15750832 TI - Tumor sensitivity to IFN-gamma is required for successful antigen-specific immunotherapy of a transplantable mouse tumor model for HPV-transformed tumors. AB - PURPOSE: Many human tumors lose responsiveness to IFN-gamma, providing a possible mechanism for the tumor to avoid immune recognition and destruction. Here we investigate the importance of tumor responsiveness to IFN-gamma in the successful immunotherapy of TC1 tumors that were immortalized with human papillomavirus proteins E6 and E7. METHODS: To investigate the role of IFN-gamma in vivo, we constructed a variant of TC1, TC1.mugR, that is unresponsive to IFN-gamma due to overexpression of a dominant negative IFN-gamma receptor. RESULTS: Using recombinant Listeria monocytogenes that express HPV-16 E7 (Lm-LLO-E7) to stimulate an antitumor response, we demonstrate that sensitivity to IFN-gamma is required for therapeutic efficacy in that Lm-LLO-E7 induces regression of TC1 tumors but not TC1.mugR. In addition, we show that tumor sensitivity to IFN-gamma is not required for inhibition of tumor angiogenesis by Lm-LLO-E7 or for trafficking of CD4+ and CD8+ T cells to the tumor. However, it is required for penetration of lymphocytes into the tumor mass in vivo. CONCLUSIONS: Our findings identify a role for IFN-gamma in immunity to TC1 tumors and show that loss of tumor responsiveness to IFN-gamma poses a challenge to antigen-based immunotherapy. PMID- 15750833 TI - Induction of specific human primary immune responses to a Semliki Forest virus based tumor vaccine in a Trimera mouse model. AB - Recombinant Semliki Forest virus (rSFV) enables high-level, transient expression of heterologous proteins in vivo, and is believed to be a superior vector for genetic vaccination, compared with the conventional DNA plasmid. Nonetheless, the efficacy of rSFV-based vaccine in eliciting human immune responses has not been tested. We used a Trimera mouse model, consisting of lethally irradiated BALB/c host reconstituted with nonobese diabetes/severe combined immunodeficiency (NOD/SCID) bone marrow plus human peripheral blood mononuclear cells (PBMCs), to characterize the in vivo immune responses against rSFV-encoded human melanoma antigen MAGE-3. MAGE-3-specific antibody and cytotoxic T lymphocyte (CTL) activity were detected by ELISA and 51Cr-release assay, respectively, and the responses were compared with those induced by a plasmid DNA vaccine encoding the same antigen. The results showed that rSFV vaccine could elicit human MAGE-3 specific antibody and CTL response in the Trimera mice, and the antitumor responses were more potent than those by plasmid DNA vaccination. This is the first report to evaluate human immune responses to an rSFV-based tumor vaccine in the Trimera mouse model. Our data suggest that rSFV vector is better than DNA plasmid in inducing protective immunity, and the Trimera model may serve as a general tool to evaluate the efficacy of tumor vaccines in eliciting human primary immune response in vivo. PMID- 15750834 TI - In vitro dendritic cell generation and lymphocyte subsets in myeloma patients: influence of thalidomide and high-dose chemotherapy treatment. AB - While vaccination with antigen-pulsed dendritic cells (DCs) represents a promising therapeutic strategy in multiple myeloma (MM), clinical benefit, so far, has been limited to individual patients. To identify potential problems with this approach, we have analyzed the influence of treatment parameters, in particular high-dose chemotherapy (HD-CTX) and thalidomide, on in vitro DC generation and peripheral blood lymphocyte subsets in MM patients. From a total of 25 MM patients, including 14 patients on thalidomide treatment and 11 after HD CTX, in vitro DC generation from peripheral blood monocytes under serum-free condition was investigated. In addition, peripheral blood lymphocyte subsets were assessed in 17 patients including 10 patients on thalidomide treatment and 9 patients after HD-CTX. Efficient in vitro generation of DCs (median 7.1x10(6)/100 ml peripheral blood; range 0.1-42.5x10(6)/100 ml peripheral blood) expressing DC typical surface markers was observed in 23 MM patients (92%), although reduced expression of CD1a, CD40, CD83, and HLA-DR was observed in patients treated with thalidomide. With respect to lymphocyte subsets, MM patients showed significantly (p<0.05) reduced B and CD4+ lymphocytes in the peripheral blood. This effect was most prominent within 6 months of HD-CTX and in patients receiving thalidomide (usually in combination with CTX). CD8+ lymphocytes were significantly increased in MM patients. Thus, despite the well-known deficiencies in their immune system, adequate numbers of DCs can be generated in most myeloma patients. In patients treated with thalidomide, however, it remains to be seen whether the reduced expression of co-stimulatory molecules has functional relevance. PMID- 15750836 TI - Sarcomeric determinants of striated muscle relaxation kinetics. AB - Ca2+ is the primary regulator of force generation by cross-bridges in striated muscle activation and relaxation. Relaxation is as necessary as contraction and, while the kinetics of Ca2+-induced force development have been investigated extensively, those of force relaxation have been both studied and understood less well. Knowledge of the molecular mechanisms underlying relaxation kinetics is of special importance for understanding diastolic function and dysfunction of the heart. A number of experimental models, from whole muscle organs and intact muscle fibres down to single myofibrils, have been used to explore the cascade of kinetic events leading to mechanical relaxation. By using isolated myofibrils and fast solution switching techniques we can distinguish the sarcomeric mechanisms of relaxation from those of myoplasmic Ca2+ removal. There is strong evidence that cross-bridge mechanics and kinetics are major determinants of the time course of striated muscle relaxation whilst thin filament inactivation kinetics and cooperative activation of thin filament by cycling, force-generating cross bridges do not significantly limit the relaxation rate. Results in myofibrils can be explained well by a simple two-state model of the cross-bridge cycle in which the apparent rate of the force generating transition is modulated by fast, Ca2+ dependent equilibration between off- and on-states of actin. Inter-sarcomere dynamics during the final rapid phase of full force relaxation are responsible for deviations from this simple model. PMID- 15750837 TI - beta3-Adrenergic-dependent and -independent mechanisms participate in cold induced modulation of insulin signal transduction in brown adipose tissue of rats. AB - During cold exposure, homeothermic animals mobilize glucose with higher efficiency than at thermoneutrality. An interaction between the insulin signal transduction machinery and high sympathetic tonus is thought to play an important role in this phenomenon. In the present study, rats were exposed to cold during 8 days and treated, or not, with a beta3-adrenergic agonist, BRL37344 sodium 4-2-2 (3-chlorophenyl)-2-hydroxyethyl amino propyl phenoxy-acetic acid sodium (BRL37344), or antagonist, SR59230A 3-(2-ethylphenoxy)-[(1S)-1,2,3,4 tetrahydronaphth-1-ylamino]-(2S)-2-propanol oxalate (SR59230A), to evaluate the cross-talk between insulin and beta3-adrenergic intracellular signaling in brown adipose tissue. The drugs did not modify food ingestion, body temperature, and body weight in control and cold-exposed rats. Treatment of control rats with BRL37344 led to higher insulin-induced tyrosine phosphorylation of the insulin receptors, insulin receptor substrate (IRS)-1 and ERK, higher insulin-induced IRS 1/PI3-kinase association, and higher [Ser(473)] phosphorylation of Akt. Cold exposure alone promoted higher insulin-induced tyrosine phosphorylation of the insulin receptors, IRS-1, IRS-2, and ERK, and higher insulin-induced IRS-1 and IRS-2/PI3-kinase association. Except for the regulation of ERK, SR59230A abolished all the cold-induced effects upon the insulin signal transduction pathway. However, this antagonist only partially inhibited the cold-induced increase of glucose uptake. Thus, the sympathetic tonus generated during cold exposure acts, in brown adipose tissue, through the beta3-adrenergic receptor and modulates insulin signal transduction, with the exception of ERK. However, insulin-independent mechanisms other than beta3-adrenergic activation participate in cold-induced glucose uptake in brown adipose tissue of rats. PMID- 15750838 TI - A role of thyrotropin-releasing hormone in insulin secretion by isolated rat pancreatic islets. AB - Insulin-secreting pancreatic beta cells also express thyrotropin-releasing hormone (TRH). Although the physiological role of TRH in this localization is unclear, its participation in glucoregulation has been implied. To test this hypothesis, we blocked the last step of post-translational maturation of the TRH molecule by disulfiram, which is an active inhibitor of peptide alpha-amidation (PAM) within pancreatic islet cells. The treatment of male rats with 200 mg/kg/day of disulfiram during a 5-day period resulted in a low PAM activity, a high insulin content and its basal secretion from pancreatic islets, and the inability to release insulin in response to glucose (16.7 mM) or hypo-osmotic (30%) challenge in vitro. The addition of TRH (1 nM) to the medium during incubation restored the insulin content and both basal and glucose stimulated insulin secretions to control levels. CONCLUSION: TRH plays an important role in the mechanism of insulin secretion and its response to glucose stimulation. PMID- 15750842 TI - [Therapy of organ failure in primary extracardiac diseases]. AB - There are a number of primary extracardiac circulatory disorders leading to organ dysfunction, multi-organ dysfunction syndrome and finally to multi-organ-failure. The origin of this hemodynamic disturbances are potentially different forms of circulatory shock (septic, hypovolemic, anaphylactic or neurogenic shock) and the systemic inflammatory reaction syndrome (SIRS). The primary aim of therapy is the removal of the underlying causes and the restoration of adequate tissue perfusion by using fluids, vasopressors and inotropics. The supportive treatment of the individual organ dysfunction is - at present - unequally efficient. PMID- 15750844 TI - [Unspecific abdominal complaints: diagnosis with szintigraphy]. AB - This case report describes a 65-year old man with symptomatic abdominal and pelvic splenosis. 50 Years ago he has had splenectomy after a road traffic accident. Ultrasound and CT revealed a bulky mass in the iliac fossa and multifocal intraabdominal nodules suggesting peritoneal carcinomatosis. Scintigraphy with Tc-99m-labelled heat damaged red blood cells led to definitive diagnosis. RBC-scintigraphy is a specific, easy performable diagnostic tool. It is cost-effective and can avoid other diagnostic procedures or explorative surgery. PMID- 15750843 TI - [Cardiopulmonary monitoring in gastroenterological and renal emergencies]. AB - Predominantly elderly and multimorbide patients require frequently intensive care observation and treatment due to acute gastrointestinal and renal disease. Manifest circulatory and rhythm instability, acute heart failure and severe metabolic or electrolyte derangements present indications for submission to a critical care unit. Stabilization of vital functions, control of specific therapeutic procedures (e. g. renal replacement therapy), and early recognition of secondary complications belong to the tasks of intensive care. Beyond a baseline monitoring available procedures comprises a broad spectrum from pulseoxymetrie to pulmonary artery catheter monitoring depending of the need of the individual patient. PMID- 15750845 TI - Peculiarities of L: -DOPA treatment of Parkinson's disease. AB - L-Dihydroxyphenylalanine (L: -DOPA), the anti-parkinsonian drug affording the greatest symptomatic relief of parkinsonian symptoms, is still misunderstood in terms of its neurotoxic potential and the mechanism by which generated dopamine (DA) is able to exert an effect despite the absence of DA innervation of target sites in basal ganglia. This review summaries important aspects and new developments on these themes. On the basis of L: -DOPA therapy in animal models of Parkinson's disease, it appears that L: -DOPA is actually neuroprotective, not neurotoxic, as indicated by L: -DOPA's reducing striatal tissue content of the reactive oxygen species, hydroxyl radical (HO(*)), and by leaving unaltered the extraneuronal in vivo microdialysate level of HO(*). In addition, the potential beneficial anti-parkinsonian effect of L: -DOPA is actually increased because of the fact that the basal ganglia are largely DA-denervated. That is, from in vivo microdialysis studies it can be clearly demonstrated that extraneuronal in vivo microdialysate DA levels are actually higher in the DA-denervated vs. the intact striatum of rats - owing to the absence of DA transporter (i.e., uptake sites) on the absent DA nerve terminal fibers in parkinsonian brain. In essence, there are fewer pumps removing DA from the extraneuronal pool. Finally, the undesired motor dyskinesias that commonly accompany long-term L: -DOPA therapy, can be viewed as an outcome of L: -DOPA's sensitizing DA receptors (D(1)-D(5)), an effect easily replicated by repeated DA agonist treatments (especially agonist of the D(2) class) in animals, even if the brain is not DA-denervated. The newest findings demonstrate that L: -DOPA induces BDNF release from corticostriatal fibers, which in-turn enhances the expression of D(3) receptors; and that this effect is associated with motor dyskinesias (and it is blocked by D(3) antagonists). The recent evidence on mechanisms and effects of L: -DOPA increases our understanding of this beneficial anti-parkinsonian drug, and can lead to improvements in L: DOPA effects while providing avenues for reducing or eliminating L: -DOPA's deleterious effects. PMID- 15750846 TI - Signal transduction through the RhoA/Rho-kinase pathway in smooth muscle. PMID- 15750847 TI - Myosin light chain isoform expression among single mammalian skeletal muscle fibers: species variations. AB - Extensive heterogeneity in myosin heavy chain and light chain (MLC) isoform expression in skeletal muscle has been well documented in several mammalian species. The initial objective of this study was to determine the extent of heterogeneity in myosin isoform expression among single fibers in limb muscles of dogs, a species for which relatively little has been reported. Fibers were isolated from muscles that have different functions with respect to limb extension and limb flexion and were analyzed on SDS gels, with respect to myosin isoform composition. The results of this part of the study indicate that there are at least four distinct fiber types in dog limb and diaphragm muscles, on the basis of MLC isoform expression: conventional fast (expressing fast-type isoforms of MLC1 (MLC1F) and MLC2 (MLC2F), plus MLC3), conventional slow (expressing slow type MLC1 (MLC1S) and MLC2 (MLC2S)), hybrid (expressing MLC1S, MLC1F, MLC2S, MLC2F and MLC3) and a second slow fiber type, designated as S1F. S1F fibers express MLC1F, along with MLC1S and MLC2S and relatively low levels of MLC3. The fraction of slow fibers that are S1F fibers varies among dog limb muscles, being greater in limb extensors than flexors. Furthermore, the mean level of MLC1F in S1F fibers is greater in extensors than flexors (mean levels range from approximately 3% to 50% of total MLC1). The study was, therefore, extended to include six additional species, spanning a broad range in adult body size to more thoroughly characterize heterogeneity in MLC isoform expression among mammals. The results indicate that there are distinct patterns in MLC isoform expression among fast and slow fibers among different species. Specifically, large-size mammals have two distinct types of slow fibers, based upon MLC isoform composition (conventional and S1F fibers), whereas small mammals exhibit variations in MLC isoforms between different types of fast fibers, including a fast fiber type that expresses MLC1S (designated as F1S fibers). S1F fibers were absent in rodent muscles and F1S fibers were not found in large mammals. We conclude that extensive variation exists in MLC isoform expression in mammalian skeletal muscle fibers, yet there are distinct patterns among different species and among muscles within an individual species. PMID- 15750848 TI - Single particle analysis: a new approach to solving the 3D structure of myosin filaments. AB - Knowledge of the structure of muscle myosin filaments is essential for a proper understanding of sarcomere structure and how myosin heads interact with the actin filaments to produce force and movement. Two principal methods have been used to define the myosin head arrays in filaments in the relaxed state, namely modelling from low-angle X-ray diffraction data and image processing of electron micrographs of isolated filaments. Analysis of filament images by 3D helical reconstruction, which imposes total helical symmetry on the structure, is very effective in some cases, but it relies on the existence of very highly ordered preparations of straight filaments. Resolutions achieved to date are about 70 angstroms. Modelling of X-ray diffraction data recorded from whole relaxed fish or insect muscles has also been used as an independent method. Although the resolution of the diffraction data is often also about 70 angstroms, the effective resolution of the modelling is very much higher than this because additional very high resolution data (e.g. from protein crystallography) is included in the analysis. However, the X-ray diffraction method has to date provided only limited data on non-myosin thick filament proteins such as C protein and titin and it cannot provide the polarity of the myosin head arrangement. Both the helical reconstruction and X-ray diffraction techniques have advantages and disadvantages, but their disadvantages are avoided in the new approach of single particle analysis of electron micrograph data. Even using the same micrographs as for helical reconstruction, the resolution can be extended by this method to about 50 angstroms or better. In addition, it is not necessary to assume that the myosin filaments are helical; a significant advantage in the case of vertebrate myosin filaments where there is a known crossbridge perturbation. Here we describe the principles of all these approaches, but particularly that of single particle analysis. We outline the application of single particle analysis to myosin filaments from vertebrate skeletal and insect flight (IFM) muscle myosin filaments. PMID- 15750849 TI - Involvement of c-Jun N-terminal kinase activities in skeletal muscle differentiation. AB - Previous studies on skeletal muscle differentiation showed that myogenesis is regulated by extracellular signal-regulated kinases (ERK-1/-2) and p38 mitogen activated kinase (MAPK) pathways. Present study shows that c-Jun NH2-terminal protein kinase (JNK) activities were up regulated during skeletal muscle differentiation in rat skeletal muscle L6E9 cells, as determined by Western immunoblot of differentiating cells probed with anti-phospho-JNK antibody. Inhibition of JNK activities by JNK inhibitor II drastically inhibited differentiation as determined by decreased myosin, myogenin expression and creatine kinase activity. The inhibition of the differentiation was regulated by apoptosis as determined by the detection of poly(ADP-ribose) polymerase (PARP) cleavage, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) positive cells when JNK activities were inhibited. Apoptosis was accompanied by marked expression and activation of c-Jun and p53 transcription factors. Taken together, our results indicate that basal JNK activities are essential for regulating skeletal muscle differentiation, and inhibition of JNK activation affects myogenesis by apoptosis dependent on c-Jun and p53 transcription factors. PMID- 15750851 TI - Conference report: fifth international conference on muscle energetics, July 2001. PMID- 15750850 TI - Telokin mediates Ca2+-desensitization through activation of myosin phosphatase in phasic and tonic smooth muscle. AB - Telokin, a 17 kDa smooth muscle specific protein, consists of the C-terminal domain of MLCK, is phosphorylated by PKA and PKG at Ser13 in vivo (Wu et al. (1998) J Biol Chem 273: 11362-11369; Walker et al. (2001) J. Biol Chem 276: 24519 24524) and is proposed to induce Ca2+-desensitization through activation of myosin phosphatase (Wu et al. (1998) J. Biol Chem 273: 11362-11369). Telokin is reported to be highly expressed in phasic with only trace amounts in tonic smooth muscle. In alpha-toxin permeabilized femoral artery, 5 microM 8-Br-cGMP induced a two-fold increase in telokin phosphorylation and a maximal 30% relaxation of Ca2+ activated force compared to a 90% relaxation in phasic ileum muscle consistent with the relative amounts of telokin expressed in ileum, 27+/-4.6 microM SEM compared to 6+/-1.7 microM SEM, in femoral artery. Recombinant Wt telokin and the phospho-telokin mutant, S13D relaxed telokin-depleted femoral artery, by 38+/-8% SEM and 60+/-20% SEM, respectively. 8-Br-cGMP increased the rate and decreased the amplitude of force development initiated by photolysis of caged ATP in alpha toxin permeabilized ileum and femoral artery smooth muscle, consistent with a cGMP-induced increase in phosphatase activity. Similarly, in telokin depleted ileum, recombinant S13D mutant telokin significantly increased the rate (0.08+/ 0.01 s-1 vs. 014+/-0.02 s-1) and decreased force amplitude. In conclusion, our data support a role for telokin in cyclic nucleotide-induced relaxation of not only phasic, but also tonic smooth muscle and that this relaxation is mediated by activation of myosin phosphatase activity leading to a decrease in myosin light chain phosphorylation. PMID- 15750852 TI - Cancer disparities: developing a multidisciplinary research agenda - preface. AB - In this Special Issue, we present a series of papers concerned with developing a multidisciplinary research agenda to address social disparities in cancer. Motivating this Special Issue are two concerns: (1) despite major advances in knowledge during the 20th century about the extent, determinants, treatment, and prevention of cancer, social disparities in cancer, including within the United States, remain serious, persistent, and require redress; and (2) huge gaps in knowledge exist regarding the causes of and solutions to social disparities in cancer across the full cancer continuum, from prevention to occurrence to diagnosis to treatment and, all too often, to death. Consequently, critical research is critically needed, both to answer the unanswered questions and to ascertain why existing knowledge is not implemented to reduce and eliminate these disparities. To help advance work in this field, we share papers developed for a January 2004 workshop on cancer disparities held at the Dana Farber/Harvard Cancer Center (DF/HCC), in Boston, MA. Topics addressed include: the definition and magnitude of, and programmatic responses to, social disparities in cancer, plus development of a systematic approach to raising research questions to address cancer disparities, using the case examples of breast, cervix, colon, and prostate cancer. PMID- 15750853 TI - Defining and investigating social disparities in cancer: critical issues. AB - Research and action to address social disparities in cancer requires clarity about what constitutes and causes these persistent and onerous inequities in health. Currently, both scientific literature and government documents exhibit important disagreements, confused terminology, and considerable, if not deliberate, vagueness about the meaning of the phrase 'cancer disparities' and the related term 'social disparities in health.' This article accordingly reviews critical issues relevant to cohering understanding of what is meant by 'cancer disparities;' offers a definition premised on the causal contention that social disparities in health, by definition, arise from social inequity; and considers its implications for developing a multidisciplinary research agenda on social inequalities in cancer. Tackling this issue will require rigorous and critical frameworks, questions, and methods derived from multiple disciplines, and will necessarily involve epidemiologic, clinical, and intervention research, both quantitative and qualitative. At issue is making conscious research choices: about which types of disparities we study, in relation to which aspect of cancer, so as to improve the likelihood our research will help inform a society-wide discourse about the extent, origins of, and remedies for social injustices in cancer, thereby aiding efforts to eliminate social inequalities in health. PMID- 15750854 TI - Using public health data systems to understand and eliminate cancer disparities. AB - Identifying and eliminating social disparities in cancer depend upon the availability and ready use of public health surveillance data at the national, state and local levels. As an example of advancing a statewide research agenda in cancer disparities, we present descriptive statistics from major public health surveillance data systems in Massachusetts. Disparities highlighted include higher breast cancer mortality rates among African-American women than women of other racial groups, lower rates of colorectal and cervical cancer screening among Asian-American residents, and striking gradients in cancer risk factor prevalence and screening by income and education. Challenges in utilizing public health surveillance data include lack of information in many domains of social inequity beyond race/ethnicity, uneven quality, and lack of stable, reportable data for smaller populations. Opportunities to maximize the usefulness of cancer registry data include application of geographic information systems and linkage with other data systems tracking information on health services outcomes and clinical trial participation. Analyses of surveillance data can spark advances not only in community-based participatory research but also in programs and policies that may ultimately eliminate disparities along the cancer continuum. PMID- 15750855 TI - The effects of a National Breast and Cervical Cancer Early Detection Program on social disparities in breast cancer diagnosis and treatment in Massachusetts. AB - OBJECTIVE: To assess social disparities in breast cancer diagnosis and treatment by comparing the stage at diagnosis and treatment of women diagnosed with breast cancer through a National Breast and Cervical Cancer Early Detection Program (NBCCEDP) for low income and uninsured women in Massachusetts, the Women's Health Network (WHN), to other breast cancer patients in the state. METHODS: We linked data from the WHN and the Massachusetts Cancer Registry (MCR). We compared 331 WHN women and 13,372 other breast cancer patients in Massachusetts diagnosed from 1995 to 1999. We used logistic regression, controlling for age, race/ethnicity, marital status, region of residence, and stage, where appropriate. RESULTS: Compared to other breast cancer patients reported to the MCR, WHN women were just as likely to be diagnosed at an advanced stage (III or IV), treated with surgery, chemotherapy, and hormone therapy, and treated with complete mastectomy versus partial mastectomy. WHN women were less likely to receive radiation therapy (odds ratio = 0.7; 95% confidence interval = 0.6-0.9), particularly after partial mastectomy, and had a slightly longer time from diagnosis to treatment than other breast cancer patients (p < 0.01). CONCLUSIONS: Women diagnosed with breast cancer through a NBCCEDP in Massachusetts had similar stage and treatment patterns as other breast cancer patients in the state, except for the use of radiation therapy. PMID- 15750856 TI - Disparities across the breast cancer continuum. AB - OBJECTIVE: We performed a structured review of the literature to identify areas of greater and lesser knowledge of the nature of disparities across the breast cancer continuum from risk and prevention to treatment and mortality. METHODS: We searched OvidMedline and PubMed to identify published studies from January 1990 to March 2004 that address disparities in breast cancer. We read the abstracts of the identified articles and then reviewed the articles if they were in English, were limited to American populations, limited to women, and described quantitative outcomes. We designated the articles as addressing one or more disparities across one or more of the domains of the breast cancer continuum. RESULTS: Substantial research exists on racial disparities in breast cancer screening, diagnosis, treatment, and survival. Disparities in screening and treatment exist across other domains of disparities including age, insurance status, and socioeconomic position. Several gaps were identified including how factors interact. CONCLUSION: A structured review of breast cancer disparities suggests that research in other domains of social inequality and levels of the cancer continuum may uncover further disparities. A multidisciplinary and multi pronged approach is needed to translate the knowledge from existing research into interventions to reduce or eliminate disparities. PMID- 15750857 TI - Social disparities and prostate cancer: mapping the gaps in our knowledge. AB - To evaluate the current state of our knowledge regarding social disparities and prostate cancer and to map the domains where substantial knowledge has been acquired as well as those where little is known, with the purpose of identifying important areas for future research. A Medline research was conducted to identify published papers regarding social disparities in prostate cancer since 1990. The results of this review are presented in a social disparities and prostate cancer grid designed to highlight which domains of social disparities have been researched and which neglected. The major social disparity in prostate cancer concerns the extremely high prostate cancer incidence and mortality seen among black Americans. This is also the area where the most research has been performed. Low socioeconomic position is associated with poorer prostate cancer outcomes but not with higher prostate cancer incidence. It remains poorly defined to what extent racial/ethnic differences in prostate cancer result from differences in socioeconomic position (SEP). Understanding the causes of the high prostate cancer mortality seen among black men remains the major challenge in the area of social disparities and prostate cancer. PMID- 15750858 TI - Social disparities across the continuum of colorectal cancer: a systematic review. AB - OBJECTIVE: The purpose of this review is to evaluate the published literature to assess social inequalities in colorectal cancer using the 'cancer disparities grid.' METHODS: Three computerized databases were searched from January 1990 to January 2004 to identify published English language articles that collected data from study participants living in the United States. Abstracts were reviewed and articles that dealt with social inequality and colorectal cancer were selected. A total of 46 articles were identified and classified into the appropriate cell of the cancer disparities grid. RESULTS: The majority of research identified for the grid has focused primarily in one domain of inequality, race/ethnicity and racism, and within one column of the cancer continuum, cancer screening. About one-third of the articles focused on multiple aspects of social inequalities. There were few or no published research articles within many of the domains of social inequality along the continuum of colorectal cancer prevention, treatment, and outcomes. CONCLUSIONS: This review found only a modest amount of research has been conducted that has examined the influence of social inequalities on colorectal cancer. Findings suggest that a multidisciplinary approach is needed to measure and remedy these social inequalities. PMID- 15750859 TI - Social inequities along the cervical cancer continuum: a structured review. AB - OBJECTIVE: To reveal areas of research/knowledge related to social inequities and cervical cancer. METHODS: A Medline search was performed looking for US based research on cervical cancer and social inequities since 1990. The papers found were organized into cells defined by a "cancer disparities grid." RESULTS: The majority of research published about cervical cancer and social inequities in the US, lies within the social domains of: race/ethnicity and socioeconomic position. Conflicting information exists as to whether race/ethnicity is a good predictor of screening and survival. Some research implied that differentials based on race/ethnicity are likely secondary to differentials in socioeconomic position. Some research about age, insurance status, and immigrant status and cervical cancer was found. Scarce information was found relating to sexuality, language, disability and geography and cervical cancer. DISCUSSION: The "cancer disparities grid" facilitated a systematic and visual review of existing literature on social inequities and cervical cancer. The grid helped to elucidate uncontested existing social inequities, conflicting social inequities, and areas where social inequity data does not exist. The cancer disparities grid can be used as a research tool to help identify areas for future research, clinical programs, and political action related to cervical cancer and social inequities. PMID- 15750860 TI - Social disparities in cancer: lessons from a multidisciplinary workshop. AB - The problem of social disparities in cancer continues to challenge health care researchers. This article summarizes the themes and lessons emerging from a 2004 workshop that convened researchers from academic and government venues to review and discuss the extant literature, and to develop new conceptual frameworks for future investigations. Workshop participants explored the factors that contribute to social inequalities in cancer in the U.S. including the relative contributions of race and racism, the independent contributions of socioeconomic position, insurance, and access to care. Noting the heterogeneous patterns of inequality across cancer types, the multiple underlying causes of disparities, and the role of the health care system itself, the authors call for an organized program of multidisciplinary research. PMID- 15750861 TI - Characterization of a vaccinia virus strain used to produce smallpox vaccine in Argentina between 1937 and 1970. AB - Due to recent political developments, smallpox has re-emerged as a serious threat. We recovered and characterized an old batch of smallpox vaccine, Malbran strain, produced between 1945 and 1949. The virus was re-isolated and characterized by sequence analysis and biological activity in animals. Phylogenetic analysis using the hemagglutinin and A45R genes showed that the Malbran strain was closely related to the Lister strain of vaccinia virus. In animals, the Malbran strain exhibited low pathogenicity, confirming historical records. Mice immunized with the Malbran strain survived a lethal challenge with cowpox virus. Thus, this strain of vaccinia virus remains a viable candidate as a smallpox vaccine. PMID- 15750862 TI - Characterization of T-cell reactive epitopes in glycoprotein G of herpes simplex virus type 2 using synthetic peptides. AB - We have previously shown that the CD4+ T-cell response to herpes simplex virus type 2 glycoprotein G-2 is type-specific and can thus be used to evaluate herpes simplex virus type 2-specific T-cell responses in individuals with a concomitant herpes simplex virus type 1 infection. In this study we have followed the glycoprotein G-2-specific T-cell responses over time, and also tried to identify T-cell epitopes in the membrane bound portion and the secreted portion of glycoprotein G-2 using synthetic peptides spanning the whole amino acid sequence of glycoprotein G-2. We found that the magnitude of the glycoprotein G-2-specific response varied considerably in infected individuals over time, even though all patients responded to at least one of the two glycoproteins at all time-points examined. We could also document strong T-cell responses to synthetic peptides from the secreted glycoprotein G-2 but only low responses to synthetic peptides corresponding to sequences from the heavily glycosylated membrane-bound glycoprotein G-2. We were able to map an immunogenic region (amino acid 31-125) within the secreted glycoprotein G-2. This region of the glycoprotein induced proliferative responses in 47% of the herpes simplex virus type 2-infected individuals. However, we were not able to identify any universal T-cell epitope. PMID- 15750863 TI - Attachment of bovine parvovirus to O-linked alpha 2,3 neuraminic acid on glycophorin A. AB - The bovine parvovirus (BPV) hemagglutinates human erythrocytes by binding to glycophorin A (GPA). The purpose of this study was to determine which carbohydrate on GPA binds BPV. Treatment of GPA with alpha2,3,-6,-8 neuraminidase eliminated binding of BPV to GPA. Beta-elimination of O-linked sialic acids on GPA eliminated binding, while removal of N-linked carbohydrates using the N glycosidase PNGase F failed to eliminate binding. Treatment of GPA with a neuraminidase which specifically cleaved alpha2,3 glycosidic bonds eliminated BPV binding and, following this treatment, virus binding to GPA was restored by reconstitution of alpha2,3-linked neuraminic acids. These results indicated the O linked alpha2,3 neuraminic acids of GPA bind BPV. PMID- 15750864 TI - Two unique US isolates of Pepino mosaic virus from a limited source of pooled tomato tissue are distinct from a third (European-like) US isolate. AB - Three strains of Pepino mosaic virus (PepMV) found in the US have been cloned and sequenced by RT-PCR using total RNA from infected tissue as template, and degenerate potexvirus- and PepMV species- and isolate-specific primers. Despite limited source material, the complete nucleotide sequences (6413 and 6410 nts, respectively) of two isolates, PepMV-US1 and PepMV-US2, were obtained and analyzed using total RNA from less than 0.2 g of a pooled infected tomato leaf sample from Arizona. Sequence of the 3'-end of the third isolate from infected fresh tomato fruits from Maryland (PepMV-US3) was also determined. The genome organizations of PepMV-US1 and US2 were typical of the genus Potexvirus, with the following reading frame order: ORF 1, encoding a putative replicase; ORFs 2-4, triple gene block proteins (TGBp) 1-3; and ORF 5, coat protein (CP). Gene-for gene comparison between PepMV-US1 and US2 revealed the following amino acid identities: 91% in replicase, 89% in TGBp1, 92% in TGBp2, 85% in TGBp3, and 93% in the CP; with an overall nucleotide identity of 86%. Nucleotide sequence comparisons between US1 and US2 and the European isolates showed only 79-82% identity, whereas the identity among the European isolates was over 99%. Sequence comparisons and phylogenetic analysis indicate that PepMV-US1 and US2 are distinctly different from the European isolates, while the CP of PepMV-US3 is nearly identical to the European isolates. The results presented also suggest that TGBp1 and TGBp3 are more suitable than either the replicase or coat protein gene products for discriminating PepMV isolates. PMID- 15750865 TI - Molecular characterization of a partitivirus from the plant pathogenic ascomycete Rosellinia necatrix. AB - The W8 isolate of the phytopathogenic fungus, Rosellinia necatrix that causes white root rot, contained three segments of double-stranded (ds) RNA, namely L1, L2 and M. Purified viral particles of about 25 nm in diameter contained an RNA segment with almost the same mobility as M-dsRNA, but the band was sensitive to S1 nuclease. Molecular analysis revealed that M-dsRNA consisted of two (RNA 1 and RNA 2) similarly sized species of 2299 and 2279 bp excluding an interrupted poly (A or U) tail of 16-51 bp. The predicted largest open reading frame in RNA 1 and RNA 2 was similar to those of RNA dependent RNA polymerase (RdRp) and coat protein (CP), respectively, encoded by the family Partitiviridae. The non-coding regions (NCR) of the two segments were similar (approximately 70% base identity) at the 5' end, but different at the 3' end. The NCR at the 3' end contained adenosine-uracil rich elements (AREs) in both segments. Northern analyses revealed RNA 1 and RNA 2 in mycelial and viral particle fractions. We coined the name Rosellinia necatrix partitivirus 1-W8 (RnPV1-W8) for M-dsRNA based on viral particle morphology and sequence information. PMID- 15750866 TI - Optimum signal in a simple neuronal model with signal-dependent noise. AB - How does the information about a signal in neural threshold crossings depend on the noise acting upon it? Two models are explored, a binary McCulloch and Pitts (threshold exceedance) model and a model of waiting time to exceedance--a discrete-time version of interspike intervals. If noise grows linearly with the signal, we find the best identification of the signal in terms of the Fisher information is for signals that do not reach the threshold in the absence of noise. Identification attains the same precision under weak and strong signals, but the coding range decreases at both extremes of signal level. We compare the results obtained for Fisher information with those using related first and second moment measures. The maximum obtainable information is plotted as a function of the ratio of noise to signal. PMID- 15750867 TI - Stochastic correlative firing for figure-ground segregation. AB - Segregation of sensory inputs into separate objects is a central aspect of perception and arises in all sensory modalities. The figure-ground segregation problem requires identifying an object of interest in a complex scene, in many cases given binaural auditory or binocular visual observations. The computations required for visual and auditory figure-ground segregation share many common features and can be cast within a unified framework. Sensory perception can be viewed as a problem of optimizing information transmission. Here we suggest a stochastic correlative firing mechanism and an associative learning rule for figure-ground segregation in several classic sensory perception tasks, including the cocktail party problem in binaural hearing, binocular fusion of stereo images, and Gestalt grouping in motion perception. PMID- 15750868 TI - Involuntary retrieval in alphabet-arithmetic tasks: task-mixing and task switching costs. AB - This study explores the effects of memory retrieval in task switching. To this end, item-specific stimulus-to-task mappings were manipulated in two "alphabet arithmetic" experiments. Letter-stimuli were presented and the responses were verbal letter names. The task was either to name the next letter in the alphabet, (e.g., C --> "D," task "plus"), or to name the preceding letter (e.g., C --> "B," task "minus"). The mapping of individual stimuli to the two tasks (and thus to responses) was either consistent (CM) or varied (VM). In Experiment 1, performance was worse for VM items relative to CM items, indicating item-specific task-mapping effects. These task-mapping effects also contributed to mixing costs (i.e., worse performance in mixed-task blocks than in pure-task blocks) but not to switch costs (worse performance in task-switch trials than in repeat trials within mixed blocks). Experiment 2 manipulated pure and mixed tasks between participants, and the data again showed differential effects of the task-mapping manipulation on mixing costs and switch costs. This suggests that, in these memory-dependent, alphabet-arithmetic tasks, interference due to involuntary task (and/or response) retrieval primarily increases general multi-task effects, such as maintaining activation of the current task. PMID- 15750869 TI - On the difference between response inhibition and negative priming: evidence from simple and selective stopping. AB - Negative priming is a commonly observed after-effect in studies concerning inhibition. Effects of the preceding trial are also found in other paradigms, like the stop signal paradigm. In the present study, stop signals were introduced in a negative priming paradigm and the relation between stop signal inhibition and negative priming was investigated. In Experiment 1, we used a simple stop signal task. Stopping data clearly suggest that stopping performance was not influenced by negative priming. Interestingly, on no-signal probes the negative priming effect disappeared after successful inhibition of the response on the prime trial. On the contrary, when inhibition failed, the negative priming effect remained. In Experiment 2, we used the selective stop signal task. As in Experiment 1, inhibition of motor responses was not influenced by negative priming. The hypothesis that negative priming disappeared due to a general nonspecific stop was confirmed in this experiment, as a negative priming effect was found after both successful and unsuccessful behavioral inhibition. The results of both experiments show that response inhibition is not influenced by negative priming, and that negative priming is only affected after a successful general stop. PMID- 15750870 TI - Rapid authentication of ginseng species using microchip electrophoresis with laser-induced fluorescence detection. AB - Ginseng is one of the most expensive Chinese herbal medicines and the effectiveness of ginseng depends strongly on its botanical sources and the use of different parts of the plants. In this study, a microchip electrophoresis method coupled with the polymerase chain reaction (PCR)-short tandem repeats (STR) technique was developed for rapid authentication of ginseng species. A low viscosity hydroxypropyl methylcellulose (HPMC) solution was used as the sieving matrix for separation of the amplified STR fragments. The allele sizing of the amplified PCR products could be detected within 240 s or less. Good reproducibility and accuracy of the fragment size were obtained with the relative standard deviation for the allele sizes less than 1.0% (n=11). At two microsatellite loci (CT 12, CA 33), American ginseng had a different allele pattern on the electropherograms compared with that of the Oriental ginseng. Moreover, cultivated and wild American ginseng can be distinguished on the basis of allele sizing. This work establishes the feasibility of fast genetic authentication of ginseng species by use of microchip electrophoresis. PMID- 15750871 TI - Development of a procedure for the sequential extraction of substances binding trace elements in plant biomass. AB - This work investigates how the amounts of some important substances in a plant, and their behaviour inside the plant, depend on the levels of stress placed on the plant. To this end, model plant spinach (Spinacia oleracea L.) was cultivated on soil treated with sewage sludge. The sewage sludge contained various trace elements (As, Cd, Cu, Zn), and the uptake of these trace elements placed the plant under stress. Following this, a sequential extraction procedure was employed to determine the levels and distributions of trace elements within the most important groups of compounds present in the spinach plants. Since the usual five-step sequential extraction procedure provides only general information on the distributions of elements within individual groups of organic compounds, due to the wide range of organic compounds within the individual fractions, this scheme was extended and improved through the addition of two solvent extraction steps-a butanol step (between the ethyl acetate and methanol solvent steps) and an H(2)O step (after the methanol+H(2)O solvent step). The distributions and levels of the trace elements within the main groups of compounds in spinach biomass was investigated using this new seven step sequential extraction (water free solvents: petroleum ether (A) --> ethyl acetate (B) --> butanol (C) --> methanol (D) --> water solvents: methanol+H(2)O (1+1; v/v) (E) --> H(2)O (F) --> methanol+H(2)O+HCl (49.3+49.3+1.4; v/v/v) (G)). The isolated fractions were characterized using IR spectroscopy and the trace element contents were determined in the individual fractions. Lipophilic compounds with low contents of Cd, Cu and Zn were separated in the first two fractions (A, B). Compounds with higher As contents (11.5-12.8% of total content) were also extracted in the second fraction, B. These two fractions formed the smallest portion of the isolated fractions. Low molecular compounds from secondary metabolism and polar lipids were separated in the third (C) and fourth (D) fractions, and high molecular compounds (mainly polypeptides and proteins) separated in the fifth and sixth fractions (E, F). The addition of the H(2)O solvent step was particularly useful for separating compounds that have a significant impact on trace element bounds. The methanol fraction was dominant for all treatments, and a significant decrease in the spinach biomass separated in this fraction was observed when the soil was treated with sewage sludge. Most of the As (35.5-38.8% of total content), Cu (45.0-51.6%) and Zn (39.8-47.2%) was also determined in this fraction. The G fraction (obtained after acid hydrolysis) contained polar compounds. Most of the Cd was also found in this fraction, as was a significant amount of Zn. Non-extractable residues formed the last fraction (polysaccharides, proteins). PMID- 15750872 TI - 84th Annual Meeting Deutsche Physiologische Gesellschaft, 6-9 March 2005, Gottingen, Germany. Abstracts. PMID- 15750880 TI - [Ethical aspects of the profession of ophthalmologist]. PMID- 15750881 TI - The preocular tear film; a small but highly complex part of the eye. PMID- 15750883 TI - [The Health Institute Carlos III supports a clinical trial on triamcinolone as part of the Eye Research Network]. PMID- 15750884 TI - [Spherical aberration influence in visual function after cataract surgery: prospective randomized trial]. AB - PURPOSE: We compare the contrast sensitivity and visual acuity obtained with an anterior surface modified prolate intraocular lens (Tecnis Z9000) with the contrast sensitivity and visual acuity obtained with a standard acrylic foldable intraocular lens (Acrysof SA60 AT). METHODS: In this prospective trial, 64 patients presenting for cataract surgery were randomized to receive in both eyes either the Tecnis Z9000 intraocular lens or the the Acrysof SA60AT intraocular lens: 32 Tecnis Z9000 and 32 Acrysof SA60AT. Visual acuity and contrast sensitivity were monocular and binocularly tested in all of them preoperatively and three months after surgery. RESULTS: The Tecnis Z9000 intraocular lens provided statistically significantly better monocular (Tecnis group: 0.85; Acrysof group: 0.78; p < 0.01) and binocular (Tecnis group: 0.95; Acrysof group: 0.86; p < 0.02) visual acuity. The contrast sensitivity mean was also greater in Tecnis group in monocular and binocular conditions, but without statistical signification. CONCLUSION: Visual acuity and contrast sensitivity after cataract surgery improved in both groups. But monocular and binocular visual acuity was statistically significantly better only in Tecnis group. The contrast sensitivity mean values were greater in Tecnis group but without statistically significant differences. PMID- 15750885 TI - [Intraocular pressure measurement using the Proview tonometer]. AB - PURPOSE: To evaluate a new ocular tonometer (Proview) which functions by visualizing a phosphene after putting pressure on the upper eyelid. To ascertain its accuracy and reproducibility with respect to the Goldmann tonometer (GT). METHODS: A study on both eyes of 110 non-selected patients was performed. One measurement with GT and three subsequent measurements with Proview were taken by the same investigator. The number of failed attempts to visualize the phosphene was recorded. We evaluated each eye separately to observe the possible learning effect. RESULTS: The intraocular pressure (IOP) mean with Proview is 5 mm Hg higher than the GT (p < 0.001). The coefficient of correlation between both instruments is low (R = 0.67; R(2) = 0.45) in the right eye (RE). The difference between maximum and minimum values of the 3 measurements taken with the Proview was 3.21 (SD: 1.73) mm Hg in RE. The failed attempts to see the phosphene were 5.14 (SD: 2.39) in RE and 4.93 (SD: 2.78) in left eye (p > 0.05). CONCLUSIONS: The Proview(R) tonometer showed low accuracy and reproducibility in comparison with the GT. This tonometer requires a long learning process before phosphene visualization. The results demonstrated that this tonometer is not clinically useful, except in patients with serious corneal diseases which make measurement with GT very difficult. PMID- 15750886 TI - [Prevalence of diabetic retinopathy in the region of Girona. Study of related factors]. AB - PURPOSE: To determine of the prevalence of diabetic retinopathy in the general diabetic population in the Girona region and epidemiologic study of the related factors. METHODS: A population-based cross-sectional study of a randomised sample taken of 401 diabetics in the Girona region. Descriptive statistical analysis of the ocular lesions in relation to diabetic retinopathy and analysis of logistic regression in order to determine the related factors. RESULTS: 30.6% (IC 95%: 26.5-35.5) of the sample presented some degree of diabetic retinopathy, 19.4% (IC 95%: 15.4-24.0) at the initial stage, 8.4% (IC 95%: 5.8-1.9) pre-proliferative and 2.6% (IC 95%: 1.3-5.1) at a proliferative stage. Regarding the type of diabetes, 53% of the patients with type 1 diabetes and 34.2% of those with type 2 diabetes had retinopathy. Retinopathy was observed in 15.7% of diabetics with less than 10 years of diabetes evolution, 51.3% in those between 10 and 20 years and 62.8% in those with more than 20 years. The concentration of HbA1C was significantly associated with the degree of retinopathy. Of those patients with retinopathy, 60.3% were insulin-treated, 30.6% with oral hypoglucemiants and 10.3% on diet only. Patients with family antecedents of retinopathy presented a prevalence three times greater than those without. CONCLUSIONS: Approximately one in three diabetic patients exhibited some degree of retinopathy in this study dealing with a population-based cohort of diabetic patients in Spain. The prevalence was greater among type I diabetic patients. As expected, years of evolution of the disease, type of treatment and diabetic metabolic control were the most significant factors influencing the presence of retinopathy. PMID- 15750887 TI - [Vitreous surgery in paediatric population. Visual and anatomic outcomes]. AB - PURPOSE: To determine the anatomic and visual outcome of children that had undergone vitreoretinal surgery. METHOD: 229 clinical files where analyzed at our center, were included 73 patients, 77 eyes. We considered those who were 18 years old or younger and had undergone retinal surgery, retinopathy of prematurity (ROP) was excluded. RESULTS: The mean age was 10 years; the main indication for surgery was retinal detachment. Trauma and congenital and developmental abnormalities where found in 80% of the ocular pathology involved. 90.9% of the patients had an initial visual acuity of light perception to counting fingers. Final visual acuity ranged from counting fingers to 1.0 in 41.6%. There was no statistical significance between initial visual acuity and the final (p = 0.782) neither was the time of surgery and the final visual acuity (p = 0.454). 12.5% presented total retinal detachment and the macula was involved in 63.6% of all patients. Vitrectomy and retinopexy was the preferred surgery. Silicon oil was used in 58.4%. Retinal attachment was achieved in 63% of the patients after 6 months. CONCLUSION: In this kind of patients the ocular characteristics of the vitreous, made the surgery a very difficult procedure, and the final success include the visual rehabilitation. PMID- 15750888 TI - [Mechanisms of protein expression in the rat optic nerve. Modifications by alcohol exposure]. AB - PURPOSE: Previous work from our group demonstrated that regular high consumption of ethanol during pregnancy induces a delay in growth and structural changes in the developing eye and vision (Pinazo-Duran et al., Teratology '93; Eur J Ophthalmol '97; Stromland and Pinazo-Duran, Teratology '94; Alcohol Alcoholism '02). Our main goal is to study at a cellular and molecular level, whether or not the prenatal alcohol exposure may change the development of the glial cells and inducing the optic nerve dysmorphogenesis. We have used key protein markers to analyse the expression in the rat optic nerve throughout the pre- and postnatal periods. METHODS: To better understanding the actions of ethanol on optic nerve development in alcohol-induced and control dams, these were fed a liquid diet during gestation and lactation, containing either ethanol (5% w/, 35% of the daily food intake) or isocaloric carobydrates (35% of the daily food intake). Eyes were enucleated and processed to immunocytochemical and morphological tecnhiques and western blot approaches, using antibodies against the glial fibrillary acidid protein (GFAP), neurofilament protein (NFP) and myelin basic protein (MBP). RESULTS: Three main observations were made in the ethanol-exposed and control groups: 1) the optic nerve size was significantly lower in the ethanol group than in the control group, 2) there were statistically significant changes in optic nerve astrocytes and oligodendrocytes, optic axons and myelin sheaths and 3) a delay and altered expression of developmental proteins. CONCLUSIONS: All data support our earlier studies confirming the deleterious effects of ethanol on the developing visual system. We suggest that ethanol may alter the expression of precise genes involved in eye development and posterior remodelling. These results can be extrapolated to clinical advances in fetal alcohol syndrome and toxic optic neuropathies. PMID- 15750889 TI - [Spider's bite that develop eyelid necrosis]. AB - CASE REPORT: We report the case of a spider bite in a 7 years-old girl that develop a dermonecrosis and did not need surgical treatment. DISCUSSION: We analize the mechanisms of these kind of lesions review and update therapeutic management of these kind of lesions. PMID- 15750890 TI - [Melanocytoma of the ciliary body]. AB - CLINICAL CASE: We report the case of a 25 year old woman who attended our Hospital with a pigmented lesion in anterior chamber angle of her right eye. She complained of reduced visual acuity on the same eye. On examination, and once complementary tests were performed, a pigmented lesion located on the iris root and ciliary body of her right eye was confirmed. A decision to perform a local resection was made and there were no surgical complication. Histology results confirmed the diagnosis of ciliary body melanocytoma. DISCUSSION: Ciliary body melanocytoma is a benign rare lesion with only 40 cases described in the literature. A local resection of this benign lesion should be considered as alternative to enucleation, even though differential diagnosis with malignant melanoma must also be considered. PMID- 15750891 TI - [Positron emission tomography in the diagnosis of orbital relapse of choroidal melanoma]. AB - CASE REPORT: A 72-year-old man with a large choroidal melanoma was treated with enucleation. Three years later MRI demonstrated images compatible with recurrent tumor in the orbit. PET studies showed no focal hypermetabolic abnormalities. DISCUSSION: In patients after enucleation of choroidal melanoma, conventional imaging techniques can fail to establish differentiation between malignant from post-surgery and benign anatomic abnormalities in orbital tissue that can be detected by PET scanning. PMID- 15750892 TI - [The eye in the human body composition (1556)(I)]. PMID- 15750893 TI - [A grave eye injury]. PMID- 15750895 TI - [Relationship between memory tests and volumetry and relaxometry of the hippocampus]. AB - INTRODUCTION AND AIMS: Interdisciplinary research made by neuropsychologist, neurologists and radiologists is making possible descriptions of the anatomic bases of memory. The hippocampus is one of the main structures related to memory processing. The aim of the present study was to study the relationship between verbal and visual memory, and the volumetry and relaxometry of the. PATIENTS AND METHODS: 36 patients with temporal lobe epilepsy and mesial temporal sclerosis (MTS). Fist, a neuropsychological assessment of was made the verbal and visual memory. Second, hippocampal structure were studied by magnetic resonance image (hippocampal volumetry and relaxometry). RESULTS AND CONCLUSIONS: Reactive gliosis measured by T2 relaxation time predicted poorer visual memory in patients with right or left MTS. These results indicate that structural damage in right hippocampus implies worse performance in visual memory. Moreover, MTS was found to have negative effects on contralateral memory (poorer verbal memory in right MTS, and poorer visual memory in left MTS). These results could be due to memory compensation. Thus, the memory function associated with the sclerotic hippocampus is adopted by the contralateral structure. Therefore, the more damage there is to the latter, the poorer is the memory functioning. PMID- 15750896 TI - [Clinical and electroencephalographic aspects of late infantile neuronal ceroid lipofuscinosis]. AB - AIMS: In this paper we describe the clinical characteristics, and particularly the epileptic seizures and electroencephalographic findings, in 15 patients with a pathology diagnosis of late infantile neuronal ceroid lipofuscinosis (NCL). PATIENTS AND METHODS: Nine female and six male patients were studied and their clinical records covering the period February 1990 to June 2003 were analysed. Neuroimaging, neurometabolic studies, ERG, PE and repeated EEG were carried out in all cases. RESULTS: The mean age on onset of the disease was 3 years (range: 1 5 years). The initial symptom was epilepsy in all cases. Massive myoclonias and myoclonic-atonic seizures were the most frequent kinds of attacks. Focal myoclonias were observed in six patients. Other types of epileptic seizures observed included generalised tonic-clonic, absence, motor focal and complex focal. The epileptic seizures were resistant to therapy. Progressive neurological and visual impairment, pyramidal and cerebellar signs, as well as mental retardation were present in all cases. Intercritical EEG recordings showed diffuse paroxysms with spike and polyspike waves, multifocal spikes and, less often, focal spikes that were predominant in posterior regions. Photostimulation showed high amplitude (300-450) occipital spikes during the application of light stimulation between 1 and 8 Hz. ERG, VEP and SSEP results were pathological. Images showed signs of brain and cerebellar atrophy. Seven of the patients died between 8.5 and 11 years of age. CONCLUSIONS: Late infantile NCL must be considered in the case of a child aged between 1 and 5 years who presents seizures that are predominantly generalised myoclonias and myoclonic-atonic, in association with progressive neurological deterioration including pyramidal, cerebellar and visual signs and an EEG trace showing occipital paroxysms triggered by low frequency photostimulation. PMID- 15750894 TI - Congenital diaphragmatic hernia and chromosome 15q26: determination of a candidate region by use of fluorescent in situ hybridization and array-based comparative genomic hybridization. AB - Congenital diaphragmatic hernia (CDH) has an incidence of 1 in 3,000 births and a high mortality rate (33%-58%). Multifactorial inheritance, teratogenic agents, and genetic abnormalities have all been suggested as possible etiologic factors. To define candidate regions for CDH, we analyzed cytogenetic data collected on 200 CDH cases, of which 7% and 5% showed numerical and structural abnormalities, respectively. This study focused on the most frequent structural anomaly found: a deletion on chromosome 15q. We analyzed material from three of our patients and from four previously published patients with CDH and a 15q deletion. By using array-based comparative genomic hybridization and fluorescent in situ hybridization to determine the boundaries of the deletions and by including data from two individuals with terminal 15q deletions but without CDH, we were able to exclude a substantial portion of the telomeric region from the genetic etiology of this disorder. Moreover, one patient with CDH harbored a small interstitial deletion. Together, these findings allowed us to define a minimal deletion region of approximately 5 Mb at chromosome 15q26.1-26.2. The region contains four known genes, of which two--NR2F2 and CHD2--are particularly intriguing gene candidates for CDH. PMID- 15750897 TI - [Single fibre electromyography in 18 patients with amyotrophic lateral sclerosis]. AB - INTRODUCTION: Single fibre electromyography (SFEMG) is a method that complements conventional electromyography and studies the activity of individual muscle fibres and of the neuromuscular junction. AIMS. We attempt to demonstrate the presence of reinnervation disorders by studying Jitter, fibre density (FD) and the presence of neurogenic blocks in patients with amyotrophic lateral sclerosis (ALS). PATIENTS AND METHODS: SFEMG was carried out on 18 patients diagnosed with ALS. All the patients were submitted to a voluntary SFEMG in the extensor digitorum communis muscle of the fingers on the most affected side. RESULTS: The SFEMG in the extensor digitorum communis muscle of the fingers showed alterations in 13 out of the 18 patients studied (72%). FD was the most reliable parameter for translation of the motor neuron instability and increased in all cases. This was followed by the mean and the maximum individual (62%) Jitter and, lastly, the percentage of blocks (38%). CONCLUSIONS: In ALS, the parameters studied with SFEMG were modified in 72% of the cases studied. There is an overall correlation between the length of time the patient has had the disease and the modifications in the SFEMG (21.5 versus 13.7 months). The degree of clinical involvement is higher (6.5/13) in those who present an altered SFEMG compared to those who present a normal SFEMG (5.3/13). PMID- 15750898 TI - [Survival of dopaminergic neurons that were hibernated in vitro for seven days]. AB - INTRODUCTION: The use of fresh foetal tissue in neurotransplants entails considerable problems of logistics that limit its clinical applicability, something that can be resolved by the development of optimal tissue storage procedures that do not affect in vivo viability and survival of dopamine. AIMS. To determine whether 7 days' hibernation affects the survival of mesencephalic tissue in vitro, and to compare it to fresh tissue. MATERIALS AND METHODS: The midbrains of rats were hibernated for 1, 3, 5 and 7 days at 4 degrees C. A cellular suspension was prepared for culture throughout a 7-day period. The number of TH+ cells present in the fresh and hibernated cultures was determined. RESULTS: The morphology of the hibernated and cultured dopaminergic neurons was very similar to that of the fresh cells. Comparing the viability of the hibernated and fresh cells did not reveal any significant differences. No significant differences between the numbers of TH+ neurons were observed at any of the hibernation times. The lowest rate of TH+ cell survival was reached at seven days' hibernation. Significant differences (p < 0.05) were found between the number of TH+ neurons for fresh and hibernated tissue. CONCLUSIONS: Hibernation at 4 degrees C for up to five days guarantees the survival of TH+ cells in vitro, but it is affected by longer times. This procedure could be considered useful for preserving human tissue in clinical transplant applications. These results refer to in vitro conditions; therefore, studies must be conducted to investigate the survival and functionality of hibernated and transplanted neurons in animal models to enable us to evaluate its applicability in neurorestorative therapy. PMID- 15750899 TI - [Colombian validation of the Pittsburgh Sleep Quality Index]. AB - INTRODUCTION: Many studies establish an elevated frequency of sleep disorders. The prevalence during life is of 35%. This situation has lead to develop a great number of instruments with the intention to evaluate better these alterations. Nevertheless, the majority lacks a suitable and national validation. Buysse et al designed the Pittsburgh Sleep Quality Index (PSQI) considered the most able instrument to establish the quality of the sleep. This has been used and validated widely. Nor it exists a version validated to the Castilian for Colombia, nor similar instruments that allow making studies in sleep. AIM: Validation of the Pittsburg Sleep Quality Index in Colombia (ICSP-VC). SUBJECTS AND METHODS: Experts consensus for face validity, pilots studies for evaluating understanding of the questionnaire, with double interview method. Evaluation of internal consistency. RESULTS. Internal consistency was Cronbach's alpha = 0.78. There were different scores between subjects that mentioned some subjective sleep disturbance (U Mann Whitney z = -5.635, p = 0.000), between them that qualify as 'so bad' or not their sleep quality (U Mann Whitney z = -2.90, p = 0.0037), between they that were consuming or not, habitually hypnotics (U Mann Whitney z = -4.289, p = 0.0000) and between they that referred conciliation insomnia or not (U Mann Whitney z = -4.769, p = 0.0000). There were clinical differences in ICSP VC scores between adult older and younger (U Mann Whitney z = -1.926, p = 0.0542). There were not differences by gender. Subjects with extreme values scores had clinical differences with others. CONCLUSIONS: ICSP-VC is a valid version of PQSI useful for studying sleep disorders. PMID- 15750900 TI - [Cystinosis: an infrequent cause of distal myopathy]. AB - INTRODUCTION: Cystinosis is a hereditary disease with clinical symptoms that are caused by the accumulation of cystine crystals in different tissues. Distal vacuolar myopathy has been reported as one of its later complications. CASE REPORT: Here, we present the case of a 20-year-old male diagnosed with cystinosis at the age of 2 years, with severe renal involvement that required a transplant. The patient gradually developed weakness and atrophy of the muscles in his hands. Neurophysiological and histological studies enabled a diagnosis of distal vacuolar myopathy to be established, and electron microscopy revealed deposits of cystine crystals. CONCLUSIONS: Cystinosis must be included within the differential diagnosis of distal myopathies. Timely treatment with cysteamine could prevent the development of this complication. PMID- 15750901 TI - [Serotonin syndrome: report of two cases and review of the literature]. AB - INTRODUCTION: Serotonin is a neurotransmitter synthesized from tryptophan. It is implied in the regulation of mood, cognition, sleep cycle, synthesis of cerebrospinal fluid, and other processes. Generally, it is implied in human pathology by hypofunction. However, there is a complication of unknown incidence related to treatment with drugs that increase the stimulation of 5-HT1A serotonin receptors, called serotonin syndrome (SS). Clinically, it is characterised by the presence of a triad of mental and autonomic disorders, and motor hyperactivity. This entity has not biological markers and its diagnosis could be done verifying the proposed criteria. CASE REPORTS: Two cases of SS are presented, one of them related to the combination of risperidone and sertraline, as first report in the literature. Both cases had a favourable outcome employing support measures. CONCLUSIONS: The physiopathology, the diagnosis, the differential diagnosis, and the treatment are reviewed. We emphasize the potentially high frequency of this disorder, given the growing use of serotonin activity modifying drugs, and the typically benign course of the SS once the support measures are started. PMID- 15750903 TI - [Sensory processing could be temporally organized by ultradian brain rhythms]. AB - INTRODUCTION: Neuronal activity of sensory systems depends on input from the environment, the body and the brain itself. Various rhythms have been shown to affect sensory processing, such as the waking-sleep cycle and hippocampal theta waves, our aim in this revision. The hippocampus, known as a structure involved in learning and memory processing, has the theta rhythm (4-10 Hz), present in all behavioural states. This rhythm has been temporally related to automatic, reflex and voluntary movements, both during wakefulness and sleep, and in the autonomic control of the heart rate. On the other hand theta rhythm has been considered as a novelty detector expressing different level of attention, selecting the information and protecting from interference. DEVELOPMENT AND CONCLUSIONS: Our research is based on the hypothesis that sensory processing needs a timer to be processed and stored, and hippocampal theta rhythm could contribute to the temporal organization of these events. We have demonstrated that auditory and visual unitary discharges in guinea pigs show phase-locking to the hippocampal theta rhythm. This temporal correlation appears during both spontaneous and specific sensory stimulation evoked discharges. Neuronal discharges fluctuate between phase-locked and uncorrelated firing modes relative to the theta rhythm. This changing state depends on known and unknown situations. We have provoked, changing the visual stimuli, a power theta rhythm increment and the phase-locking between this rhythm and the lateral geniculate neurone discharge during wakefulness. In slow wave sleep results were different demonstrating that the ways of the inputs processing have changed. PMID- 15750902 TI - [Transient ischemic attack as the presenting symptom of a pulmonary arteriovenous fistula]. AB - INTRODUCTION: Transient ischemic attacks (TIA) occur as the clinical manifestation of a pulmonary arteriovenous malformation in up to 20% of cases. CASE REPORT: We report the case of a 41-year-old female with an episode of TIA lasting half an hour, with right-side hemiparesis caused by a paradoxical embolisation due to the presence of a single pulmonary arteriovenous fistula (PAVF). Complementary analytical and imaging tests (CAT scan of the head without contrast, brain RMI and intra and extracranial MR-angiography) were normal. A transthoracic echocardiogram showed the presence of a patent foramen ovale which obstructed the passage of hemoce, although it was observed in the left auricle after presumably arriving through the pulmonary arteries. Since a pulmonary vascular malformation was suspected, a computerised tomographic angiography scan of the thorax was performed and this confirmed the presence of a single PAVF in the right lung. A pulmonary arteriography was then carried out to confirm the presence of the malformation, which was later embolised. Two months later, the patient was asymptomatic and the fistula did not appear in the computerised tomographic angiography scan of the thorax. CONCLUSIONS: PAVFs can give rise to neurological conditions due to paradoxical embolisms that can produce abscesses and infarcts and/or concomitant lesions in the central nervous system. This is a certain indication of treatment of the malformation and the preferred technique is endovascular embolisation with coils, while surgery is reserved for cases in which endovascular treatment is not possible. PMID- 15750904 TI - [Advanced dementia in Alzheimer's disease and memantine]. AB - INTRODUCTION: Alzheimer's disease (AD) causes dementia with a progressive course. Until now, research has been aimed mainly at studying its early, pre-dementia and mild or mild to moderate dementia phases in an attempt to find treatments with which to cure it or at least halt its progression. It is true that the latest cognitive therapies are effective up to a point, but the fact is that many patients reach the state of advanced AD, which gives rise to a number of cognitive and behavioural disorders that entail all kinds of problems on a personal, familial and community health level. AIMS: This study reviews the progress of AD, the length of its stages, the speed at which the disease courses and patients' survival. Advanced AD is defined, its clinical features and the functional disability it causes are described, and we analyse the tests and scales that must be used to measure how the process is progressing and the effectiveness of the distinct forms of treatment used in the later phase of AD, which are different to those employed in the early stages. All this allows us to analyse the results of clinical trials carried out with memantine, an NMDA (N methyl D-aspartate) receptor antagonist. We review its pharmacological characteristics and its use in everyday practice. Lastly, we refer to the classical symptomatic treatments that are usually employed to control the frequent and intense behavioural disorders produced in the advanced phase of AD. CONCLUSIONS: The later stage of AD requires our attention because many patients reach and remain for a long time in this phase, which leads to considerable personal and social disorders. Several scales and tests have been adapted to these later phases of AD, enabling the clinician to evaluate the patient, monitor the progress of the disease and determine the effectiveness of different treatments. Memantine, the most recent drug approved for use with this disease, has proved to be effective in the treatment of patients with advanced AD. This pharmaceutical has been added to the list of well-known classical medicines, such as neuroleptic drugs, antidepressants, anxiolytic agents and others, which can be used to diminish the behavioural disorders in these patients and improve their quality of life, as well as that of their caregivers. PMID- 15750905 TI - [Chronic daily headache: II. Treatment]. AB - INTRODUCTION: The difficulty involved in treating chronic daily headache (CDH), a frequent pathology with a multifactorial aetiology, makes it a challenging disorder for clinicians. CDH is a disease that causes significant impact on patients, employers and the health system. DEVELOPMENT: The treatment of CDH is based on the implementation of three therapies. The first is to establish a pharmacological treatment, with medication such as antidepressants, antiepileptic agents, muscle relaxants, and antihypertensive and antiserotoninergic drugs, either in mono or in polytherapy. The second measure is to stop the analgesic overuse. This is achieved by suddenly withdrawing the drugs or, in a combined manner, by stopping the analgesic intake and establishing bridge therapy (steroids or triptans), which, in addition to reducing the severity and frequency of seizures and the consumption of drugs, also help to lower the occurrence and duration of withdrawal symptoms. Hospital treatment is needed in cases of patients who are refractory to multiple therapies, with frequent visits to the emergency department, in presence of associated symptoms or comorbidities, and low tolerance to medication. The third measure is to implement non pharmacological therapies, which are used to modify inappropriate behaviour as regards pain treatment, to promote proper medication consumption and to allow the patient comprehend the prognosis of the disease with and without treatment. CONCLUSIONS: Implementing these three therapies makes possible to reduce the severity, frequency and socioeconomic impact caused by headache and thereby improve patients' quality of life and capabilities. PMID- 15750906 TI - [Epidemiological clinical behaviour of brain deaths in the North-Eastern territory of Cuba throughout the years 2002-2003]. PMID- 15750907 TI - [Stiff man syndrome: a specific immunological condition affecting the central nervous system]. PMID- 15750908 TI - Iodine overload and severe hypothyroidism in a premature neonate. AB - Use of iodinated skin disinfectants in the perinatal period can result in significant iodine overload of the neonate and transient hypothyroidism. The authors report a case of unusually severe hypothyroidism requiring L-thyroxine (L T4) replacement therapy in a premature neonate after prolonged use of iodinated skin disinfectants for a complex skin lesion. Neonatal iodine overload should be minimized, and in cases with significant iodine exposure, thyroid-stimulating hormone should be monitored, especially in preterm neonates who are exquisitely sensitive to the antithyroid effects of iodine excess. PMID- 15750909 TI - Intraneural perineurioma in a child with Beckwith-Wiedemann syndrome. AB - To the authors' knowledge, this is the first report of an intraneural perineurioma in a child with Beckwith-Wiedemann syndrome. Intraneural perineurioma, previously known as localized hypertrophic neuropathy, is a rare benign peripheral nerve sheath tumor arising from perineurium. This report adds a new entity in the spectrum of tumor formation in Beckwith-Wiedemann syndrome. PMID- 15750910 TI - Prenatal diagnosis of intrathoracic stomach (gastric herniation). AB - Intrathoracic stomach is a rare and serious congenital abnormality. The anomaly may be complicated by gastric volvulus and can lead to ischemic gastric infarction in the neonate. If diagnosed antenatally, neonatal management can be planned in advance so as to reduce morbidity. This anomaly must be differentiated from the more common congenital diaphragmatic hernia, as associated pulmonary hypoplasia is common in the latter and rare with gastric herniation. We report an infant born to a mother with Marfan's syndrome with the antenatal diagnosis of intrathoracic stomach. The ultrasound and magnetic resonance imaging features of this congenital abnormality are described. A review of the literature would indicate that this is the first case report of gastric volvulus diagnosed in utero. PMID- 15750911 TI - Type IV sacrococcygeal teratoma causing urinary retention: a rare presentation. AB - Sacrococcygeal teratoma is a rare fetal neoplasm usually diagnosed antenatally and considered benign with no functional impairment. Nevertheless, fetuses with this malformation are at risk of significant perinatal morbidity and mortality, usually implying a malignant component. Management is primarily surgical, with a generally good prognosis. Herein, the authors present a rare and unusual presentation of a type IV sacrococcygeal teratoma as urinary retention in a 7-day old infant. The diagnostic workup and subsequent surgical treatment are described. The nature of the tumor and the treatment is discussed, emphasizing the need for urgent surgical treatment because of bilateral hydronephrosis that failed to resolve after inserting a urinary catheter. PMID- 15750912 TI - Left paraduodenal hernia leading to protein-losing enteropathy in childhood. AB - A 3-year-old girl presented with a protein-losing enteropathy caused by left paraduodenal hernia, as diagnosed by computed tomography and a small-bowel follow through image. The patient received surgical treatment, and her postoperative course was uneventful. Nineteen days later, the serum protein and albumin had recovered to normal levels. As well as providing the first description of left paraduodenal hernia accompanied with protein-losing enteropathy, we also review the pertinent literature. PMID- 15750913 TI - Perforated appendicitis in a child with Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is a common childhood vasculitis. Abdominal pain is a common feature of HSP, often leading to surgical consultation for evaluation of possible intussusception. Appendicitis is a rare complication of HSP, and in each of the 3 reported cases, appendectomy preceded the appearance of the purpuric rash. More often, unnecessary laparotomies are performed on patients in whom appendicitis is suspected, but who subsequently develop the characteristic purpura. This is the first reported case of appendicitis developing in a patient with the established HSP rash. This case is also the first report of perforated appendicitis in HSP. Clinical vigilance and serial physical and ultrasonographic examinations are needed to detect conditions necessitating surgery in patients with HSP. PMID- 15750915 TI - Serial transverse enteroplasty in intestinal atresia management. AB - In intestinal atresia, resection of the proximal dilated bowel segment, if long, might lead to bowel length compromise. Although tapering enteroplasty and plication are well described to avoid such a resection, the use of serial transverse enteroplasty is proposed here as an alternative in this situation. PMID- 15750914 TI - The combination of Hirschsprung's disease and achalasia. AB - The unusual combination of Hirschsprung's disease and Achalasia in one case treated by standard procedures led to the discussion about RET germ-line mutations and consequently to the speculation about higher risk for multiple endocrine neoplasia syndrome type 2-related tumors. Although a mutation could be excluded by sequence analysis in this case, the correlation of these specific diseases affords additive investigations to make sure that no further prophylactic procedures were necessary. PMID- 15750916 TI - Traumatic rupture of choledochal cyst in a child. AB - Traumatic rupture of choledochal cyst is an extremely rare disorder. The current patient is a 4-year-old boy who fell in a bathroom and suffered a blow to the abdomen. Percutaneous transhepatic cholangiography revealed pancreaticobiliary maljunction. Inflammation of the peritoneal cavity was moderate. At first look, the choledochal cyst was excised and hepaticojejunostomy was performed. At this time, a rupture approximately 2 mm in diameter was recognized at the rear surface of the inferior part of the common bile duct. PMID- 15750917 TI - Successful liver resection for biliary atresia with intrahepatic biliary cysts after Kasai procedure. AB - A 29-year-old woman with congenital biliary atresia underwent liver resection for intrahepatic biliary cyst and recurrent cholangitis long after Kasai procedure. The patient is alive with normal liver function and no episode of cholangitis after 12 months of follow-up. PMID- 15750918 TI - Testicular descent and cryptorchidism: the state of the art in 2004. AB - The understanding of testicular descent has changed much in the 20 years since the authors' laboratory began studying the mechanism. The process is now known to occur in 2 steps with different anatomy and hormonal regulation but with many still unresolved controversies. Recent advances include the recognition of acquired cryptorchidism of critical early postnatal germ cell development and the recommendation for surgery at 6 months of age. The authors still await long-term outcome studies. PMID- 15750919 TI - Thoracoscopy in infants and children: the state of the art. PMID- 15750920 TI - Health-related quality of life in adult patients with esophageal atresia--a questionnaire study. AB - BACKGROUND/PURPOSE: Long-term sequelae caused by associated anomalies or respiratory and gastrointestinal disorders are common after the repair of esophageal atresia (EA). The aim of this study was to assess the effect of these sequelae on the health-related quality of life (HRQoL) of patients with EA. METHODS: A questionnaire including a 36-item Gastrointestinal Quality of Life Index, a 15-item Respiratory Symptoms-Related Quality of Life Index, and a 36 item psychosocial survey and a symptoms query was sent to 159 patients with EA with or without tracheoesophageal fistula (TEF), who were operated on between 1949 and 1979, and to 400 healthy control subjects. A 36-item Health Survey Form was sent to patients with EA only and the results were compared with values of the general Finnish population. RESULTS: A total of 128 patients with EA (80%) and 162 control subjects (41%) returned the questionnaire. Median age [patients with EA, 38 (range, 24-54) years; control subjects, 36 (20-56) years] and sex distribution (M/F of patients with EA, 57:70; control subjects, 63:99) were comparable. Of the 128 patients, 115 (91%) had EA with a distal TEF, 8 (6%) had EA without TEF, 3 (2%) had EA with proximal and distal TEF, and 2 (1%) had TEF without EA. The types of esophageal repair were distributed as follows: primary end-to-end anastomosis (n = 112), colon interposition (n = 10), gastric tube (n = 3), thoracic skin tube (n = 1), and fistula closure only (n = 2). Thirty-eight patients had significant associated anomalies. Gastrointestinal Quality of Life Index scores did not differ statistically between patients with EA and control subjects. However, the incidence of regurgitation (17% vs 10%) and dysphagia (10% vs 2%) was significantly higher in patients with EA (P < .5). Low Respiratory Symptoms-Related Quality of Life Index was found in 10 of 128 patients with EA and in 3 of 163 control subjects (P < .05). Psychosocial survey scores and the incidence of acquired diseases did not differ between the groups (P = NS). The 36 item Health Survey Form indicated low HRQoL in 19 (15%) of patients with EA (expected value, 16%). Of the 19 patients with EA with low HRQoL, 8 (42%) had significant congenital or EA-associated diseases and 11 (58%) had acquired diseases. The type of EA or esophageal conduit was not related to HRQoL. CONCLUSIONS: Most adult survivors of EA or TEF repair have a normal quality of life. Morbidity from esophageal functional disorders and respiratory disorders with or without acquired diseases impairs HRQoL in 15% of patients with EA. PMID- 15750921 TI - Fibroblast growth factor-10 serves a regulatory role in duodenal development. AB - PURPOSE: Duodenal obstruction occurs in 1 of 6000 live births and requires urgent surgical intervention. Duodenal atresia previously has been ascribed to a developmental failure of luminal recanalization; however, the cause of duodenal atresia remains incompletely understood. Although familial intestinal atresias have been described and syndromic associations are known, no specific genetic link has been established. Fibroblast growth factor-10 (Fgf10) is a known regulatory molecule relevant to mesenchymal-epithelial interactions, and mice deficient in Fgf10 demonstrate congenital anomalies in several organ systems including the gastrointestinal tract. The authors hypothesized that Fgf10 could serve a regulatory role in establishing normal duodenal development. METHODS: Wild-type mice with beta-galactosidase under the control of the Fgf10 promoter were harvested from timed-pregnancy mothers. The expression of Fgf10 in the duodenum during development was evaluated by developing the embryos in X-Gal solution. Wild-type and mutant Fgf10(-/-) embryos were harvested from timed pregnancy mothers at 18.5 days postconception (near term) and were analyzed for duodenal morphology (Institutional Animal Care and Use Committee-approved protocol 32-02). Photomicrographs were reviewed. RESULTS: Fibroblast growth factor-10 is active in the duodenum at a late stage of development. The Fgf10(-/ ) mutants demonstrate duodenal atresia with a variable phenotype similar to clinical findings. The duodenum fails to develop luminal continuity and has proximal dilation. The phenotype occurs in an autosomal recessive pattern with incomplete penetrance (38%). CONCLUSIONS: Fibroblast growth factor-10 serves as a regulator in normal duodenal growth and development. Its deletion leads to duodenal atresia and challenges traditionally accepted theories of pathogenesis. This novel, genetically mediated duodenal malformation reflects an animal model that will allow further evaluation of the pathogenesis of this surgically correctable disease. By studying the mechanism of Fgf10 function in foregut development, the authors hope to better understand these anomalies and to explore possible therapeutic alternatives. PMID- 15750922 TI - Abdominal wall repair using a biodegradable scaffold seeded with cells. AB - BACKGROUND/PURPOSE: The repair of large abdominal wall defects is still a challenge for pediatric surgeons. Synthetic materials, however, may lead to high complication rates. This study was aimed at applying tissue-engineering methods to abdominal wall repair. METHODS: 3T3 mouse fibroblasts were expanded in vitro. In the next step, a biodegradable material--polyglycolic acid (PGA)--was actively seeded with 10(7) cells/scm of PGA scaffold. Culture medium (Dulbecco's Modified Eagle's Medium with 10% fetal bovine serum) was changed every 6 hours after seeding cells on PGA fibers. Under general anaesthesia, C57BL/6J black mice underwent creation of a 2 x 3-cm abdominal wall defect (60%-70% of abdominal surface). The defect was repaired in the experimental group with the fibroblast seeded PGA scaffold. In the first control group, the defect was covered with acellular PGA, and in the second control group, by skin closure. Animals were killed after 30 days to assess the histologic and gross findings. RESULTS: No abdominal hernia was found in animals repaired with cell-seeded and acellular scaffolds. All animals with skin closure died within 7 days. In every case, tissue-engineered construct was thicker then in controls. Histologic and gross examination revealed a good neovascularisation in tissue-engineered abdominal walls comparing to the acellular matrix. There was no intensive scar formation between abdominal wall and skin. CONCLUSIONS: Engineered soft tissue constructs can provide structural replacement of severe and large abdominal wall defects. Tissue engineering in the near future will possibly enter clinical practice. PMID- 15750923 TI - Kasai portoenterostomy--new insights from hepatic morphology. AB - BACKGROUND/PURPOSE: The aim of this paper was to investigate the mechanism of long-term biliary drainage after Kasai portoenterostomy by clinicopathologic study of hepatic morphology in explanted livers. METHODS: Explanted livers from 13 consecutive children undergoing transplantation for biliary atresia were examined in detail using a standardized protocol. Group 1 (n = 6) had no Kasai procedure before transplantation at a median age of 8 m. Group 2 (n = 4) were transplanted at a median age of 10 m after a failed Kasai portoenterostomy. Group 3 (n = 3) had a successful Kasai but required transplantation for complications of chronic liver disease at 12-14 years. Pathology findings were correlated with hepatic morphology determined by pretransplant magnetic resonance imaging. RESULTS: Large perihilar regenerative nodules (8-14 cm diameter) were observed in 2 patients after successful Kasai portoenterostomy, less well-defined perihilar nodules in group 2 patients, and no regenerative nodules in group 1. Microscopically, group 1 had diffuse biliary cirrhosis with evidence of progressive ductopenia during infancy. In group 2, perihilar regenerative nodules showed variable portal fibrosis but no cirrhosis and bile ducts were present with 68%-100% of hepatic arteries; in peripheral cirrhotic areas, bile ducts were absent in patients older than 9 m. The perihilar regenerative nodules in group 3 patients had a noncirrhotic architecture with preserved bile ducts, but the peripheral parenchyma was cirrhotic; one patient had diffuse macronodular cirrhosis. These morphologic findings correlated well with magnetic resonance images, highlighting the preservation of relatively normal perihilar liver architecture after successful Kasai portoenterostomy. CONCLUSIONS: Unoperated biliary atresia is associated with progressive intrahepatic ductopenia leading to diffuse biliary cirrhosis. Kasai portoenterostomy can result in the growth of large perihilar regenerative nodules, probably as a consequence of surviving intrahepatic ducts in this region. In some patients, long-term success after Kasai portoenterostomy may depend on hyperplasia of the perihilar liver. PMID- 15750924 TI - Optimum prednisolone usage in patients with biliary atresia postportoenterostomy. AB - PURPOSE: Prednisolone is used routinely after portoenterostomy (PE) in patients with biliary atresia (BA). The authors reviewed their patients with BA post-PE to assess prednisolone protocols. METHOD: Severity of fibrosis at PE (moderate or severe), age at PE (30-70 days), size of bile ductules in the fibrotic biliary remnant at the porta hepatis (>100 microm), and type of BA (uncorrectable type) were used as criteria for selecting 63 subjects from our patients with BA post PE. Subjects were divided into 5 groups according to prednisolone dosage: group 1, no prednisolone; groups 2 to 4, single courses of intravenous prednisolone commencing on day 7 post-PE administered in decreasing dose for 3 days each as follows: group 2, 6, 4, and 2 mg; group 3, 10, 5, and 2.5 mg; group 4, 20, 15, 10, 5, and 2.5 mg; group 5, same as group 4, but stool color was used to monitor bile excretion and a course was restarted from 20 mg whenever stools began to turn pale. If necessary, single courses were repeated until serum total bilirubin was less than 2.0 mg/dL. Protocol efficacy was assessed by comparing the number of patients who became jaundice free, the period taken to become jaundice free, and the incidence of side effects related to prednisolone. RESULTS: The number of patients who became jaundice free in the no prednisolone group (group 1, 7/12 or 58.3%) was not significantly different from the number in the single-course groups (group 2, 8/12 or 66.6%; group 3, 10/13 or 76.9%; and group 4, 11/15 or 73.3%). The number in the stool-monitored group (group 5, 10/11 or 90.9%) was significantly greater (P < .05). The mean period taken to become jaundice free in group 1 (82.6 +/- 29.1 days) was not significantly different from the single course groups (group 2, 74.5 +/- 29.3 days; group 3, 49.6 +/- 19.8 days; and group 4, 48.3 +/- 26.0 days). The mean period taken in the stool-monitored group (group 5, 33.3 +/- 6.4 days) was significantly shorter (P < .05). The number of subjects who developed cholangitis after becoming jaundice free was not significantly different (group 1, 2/7; group 2, 2/8; group 3, 2/10; group 4, 2/11; group 5, 2/10). There were no prednisolone-related complications identified in any subject. CONCLUSIONS: These results provide strong evidence that large dose prednisolone therapy with stool color monitoring of bile flow has a positive impact on the time taken for patients with BA post-PE to become jaundice free and the number of patients who remain jaundice free. PMID- 15750925 TI - Under pressure: choledochal malformation manometry. AB - PURPOSE: The cause of choledochal (cystic or fusiform) malformation is not known. A favoured hypothesis suggests that abnormal reflux of activated pancreatic secretions via a common pancreatobiliary channel may initiate mucosal injury and mural weakness leading to bile duct dilatation, at normal intraduct pressures. However, bile duct pressures in both normal or disease states are not known in such children. METHODS: Intraoperative choledochal pressure (CP) measurements were made before any other manipulation. Bile was cultured and its amylase content measured. Biochemical liver function (bilirubin, aspartate aminotransferase, gamma-glutamyl transpeptidase, and alkaline phosphatase) was measured. Data were quoted as median (interquartile range). Statistical tests were parametric, where appropriate, and P = .05 was regarded as significant. RESULTS: Twenty-five children (age 2.5 [1.25-5.91] years) with choledochal (cystic [n = 13] and fusiform [n = 12]) malformation coming to surgery were studied. Median CP was 13 (8.5-17) mm Hg. Median bile amylase was 6722 (241 18,000) IU/L. Choledochal pressure inversely correlated with bile amylase (r = 0.60, P = .001), serum aspartate aminotransferase (r = 0.46, P = .01), and log gamma-glutamyl transpeptidase (r = 0.4, P = .04) but not with bilirubin (P = .11), alkaline phosphatase (P = .20), or age (P = .11). No difference in CP, bile amylase, or liver biochemistry could be identified between the 2 biliary phenotypes. All bile cultures were sterile. CONCLUSIONS: Increased CP is inversely related to the level of bile amylase (and hence degree of the functional common channel). This suggests that obstructive stenosis at the level of the pancreatobiliary junction (but not the ampulla) may be a causal factor in a proportion of choledochal malformations. PMID- 15750926 TI - Is portal venous outflow better than systemic venous outflow in small bowel transplantation? Experimental study in syngeneic rats. AB - BACKGROUND/PURPOSE: Blood drainage of the graft into the recipient portal vein reestablishes the physiological venous outflow after small bowel transplantation (SBT). However, although this approach is likely beneficial for the host, it may be technically more demanding making portocaval venous drainage the preferred arrangement during human SBT. The aim of this study was to examine in a syngeneic model of SBT the possible benefits of portoportal anastomosis (PPA) vs portocaval anastomosis (PCA) in terms of body and organ weights and bacterial translocation. METHODS: Syngeneic SBT was carried out in 25 Brown-Norway male rats weighing 249 +/- 17.5 g using either PPA (n = 13) or PCA (n = 12). Half the animals in each group were killed, respectively, on postoperative day 2 or 7. Liver, spleen, and lungs were weighed and under sterile conditions the regional lymph nodes were excised. The nodes and venous samples from the cava and portal veins were cultured for aerobes and anaerobes. Bacterial components were detected in blood by polymerase chain reaction. The findings in both groups were compared by chi2 or Mann-Whitney U tests. RESULTS: Mean postoperative body weight change was -3.6% +/- 1.5% in PPA and -6.0% +/- 1.2% in PCA animals (ns) on day 2 and -6.5% +/- 2.6% and -8.0% +/- 5.0% (ns) on day 7. Liver, spleen, and lung weights were not significantly different between both groups on either end point. Gram-negative enteric bacteria were found in 3 of 7 PCA animals and 2 of 6 PPA animals at day 2 (ns) and in 1 of 6 and 4 of 6 on day 7 (ns). Aerobic gram-positive bacteria were found in 1 of 7 and 1 of 6 (ns), 3 of 6 and 3 of 6 (ns), respectively, in the 4 groups. Most positive cultures corresponded to portal blood and lymph node samples. There were no anaerobic growths. CONCLUSIONS: -No body or organ weight change suggesting significant functional advantages of one technical alternative over the other could be demonstrated. -Bacterial translocation in the absence of rejection was frequent after SBT independently of the variety of venous outflow used. No difference in bacterial translocation between both anastomosis could be demonstrated. -Orthotopic venous drainage did not seem to be advantageous in the present experimental setting. PMID- 15750927 TI - Decreased E-cadherin expression correlates with higher stage of Wilms' tumors. AB - PURPOSE: The aim of this study was to examine the association between E-cadherin expression and markers of Wilms' tumor aggression, including metastasis and recurrence. METHODS: Forty Wilms' tumor samples from the National Wilms' Tumor Study Group underwent immunohistochemical staining for E-cadherin. Tumor stage at diagnosis, recurrence, and loss of heterozygosity at 16q status was known for each of the tumor samples. E-Cadherin cell staining was defined as high (>33%) or low (<33%), and values were assigned by a pathologist blinded to the tumor characteristics. Five stage IV tumors were ineligible for assay because of lack of a tubular component. To identify a mechanism of downregulation, we screened tumor DNA for genetic mutations in exons 1-16 using a combination of WAVE and sequence analysis. To assess the functional significance of the identified mutations, the authors compared amino acid homology across multiple species. Finally, they performed reverse transcriptase-polymerase chain reaction for those tumors with intronic single nucleotide polymorphisms (SNPs) to evaluate for mRNA splice variants. RESULTS: Wilms' tumors presenting with metastatic (stage IV) disease demonstrated decreased levels of E-cadherin expression compared with localized tumors (stage I) (Fisher's Exact test, P < .01). In a search for the mechanism of the downregulation of E-cadherin, we identified 5 different mutations in 7 high stage tumors (7/15) and 1 mutation in a low stage tumor (1/20). The mutations occurred in amino acids that were conserved across multiple species. Additionally, 11 of 15 high stage tumors contained an intronic SNP located within 6 bp of the 5 intronic splice junction immediately downstream of exon 1. However, examination of 5 of these tumors using reverse transcriptase polymerase chain reaction showed that this intronic SNP does not appear to disrupt the assembly of full-length E-cadherin transcripts. Lastly, no correlation was identified between E-cadherin expression and recurrence of disease. CONCLUSIONS: In this study, the authors have found an association between decreased E-cadherin expression and metastatic Wilms' tumor. Mutations identified may help identify a mechanism for downregulation. The functional significance of these mutations is supported by the conserved nature of the amino acids across multiple species. The authors believe these findings support the involvement of E-cadherin in the evolution of Wilms' tumor. PMID- 15750928 TI - Profiling of nuclear extract proteins from human neuroblastoma cell lines: the search for fingerprints. AB - PURPOSE: Neuroblastoma (NB) commonly presents with advanced disease at diagnosis and is associated with poor survival. If identified early, however, survival is improved suggesting a benefit of early detection. The authors have used proteomics technology in an attempt to identify novel markers that permit early detection of NB and characterize its molecular makeup. METHODS: Three different human NB cell lines SK-N-AS, SK-N-DZ, and SK-N-FI were subjected to series of biochemical fractionation steps to extract nuclear proteins. These proteins were analyzed for differential expression by 2-dimensional polyacrylamide gel electrophoresis. Polypeptides of interest were subsequently identified by liquid chromatography-linked tandem mass spectrometry. RESULTS: Multiple proteins were identified in these human NB cell lines including SET (a ubiquitous nuclear protein), stathmin (a cytosolic signal transduction protein), and grp94 (a heat shock protein). SET is a putative oncogene associated with the chromosomal translocation (6;9) leading to acute undifferentiated leukemia. Stathmin is an oncogene found in greater abundance in leukemic cells compared to nonleukemic cells. A total of 94-kDa glucose-regulated protein has been shown to be protective in human breast cancer cells in vitro and related with the occurrence, differentiation, and progression of human lung cancer. The first protein has not been previously associated with NB. CONCLUSIONS: The identification of these 3 previously unrecognized cancer-related potential biomarkers in human NB cell lines may prove useful in developing diagnostic tests. The proteomic methodology of 2-dimensional polyacrylamide gel electrophoresis/mass spectrometry also provides an improved opportunity to understand the natural history of NB and develop novel chemotherapeutic agents for this prevalent childhood malignancy with a dismal outcome. PMID- 15750929 TI - Profile of neuroblastoma detected by mass screening, resected after observation without treatment: results of the Wait and See pilot study. AB - BACKGROUND/PURPOSE: Neuroblastoma (NB) detected by mass screening (MS) usually shows favorable prognosis and sometimes regresses spontaneously. Therefore, the authors started an observation program for these patients to avoid overtreatment. In this study, the authors analyzed the profile of NB resected after observation to elucidate the nature of NB detected by MS. METHODS: Between 1994 and 2004, 22 NB patients matched the following criteria and entered the observation program after obtaining informed consent: stage I or II, less than 5 cm in diameter, and without involvement of large vessels or organs. If increase in size, elevation of tumor markers, or evidence of metastasis was observed, the tumor was immediately resected. RESULTS: Thirteen (59%) of 22 cases showed spontaneous regression. In the remaining 9 cases, tumors were resected because of parents' request, increase in size, and/or elevation of tumor markers. Four tumors had at least one unfavorable biologic feature, and 3 of them had more than 2. According to Shimada's system, 2 had unfavorable histology. One was diploid tumor, 3 had 1p deletion, and Trk-A expression was low in 4 tumors. All patients survived without evidence of recurrence. CONCLUSIONS: The observation program has shown that at least one third of the NB detected by MS regressed spontaneously. On the other hand, MS may detect some cases with unfavorable tumor in early stage, which benefit from screening. PMID- 15750930 TI - Three-dimensional visualization and virtual simulation of resections in pediatric solid tumors. AB - PURPOSE: Three-dimensional visualization of solid tumors is possible because of high-resolution computed tomography and magnetic resonance imaging scans. However, additional preoperative information is often desirable in complex malignancies. For the first time, the authors present a model of preoperative 3 dimensional visualization and virtual resections in pediatric solid tumors. METHODS: Image analysis of various pediatric tumors was performed using the research software HepaVision2 (MeVis, Bremen). Organs, tumors, and the vascular system were extracted from multislice computed tomography scans. After hierarchical analysis of the vascular system, territories supplied or drained by the major vascular branches were calculated. Results were explored and virtual resections of organs were carried out using the research software InterventionPlanner (MeVis, Bremen). Data were correlated to intraoperative findings. RESULTS: Four hepatic malignancies, 4 renal tumors, and 3 other neoplasms were analyzed. The technique of 3-dimensional visualization was feasible for all investigated children (mean age 5 years and 9 months). Spatial relations between physiological and pathological structures were identified, and anatomical structures (vessels, tumor tissue, and organ parenchyma) were determined using colorimetric encoding. Virtual simulations of tumor resection were used successfully for planning of surgical procedures in the hepatic and renal tumors. CONCLUSIONS: The technique of 3-dimensional tumor visualization and virtual simulation of tumor resections provides the basis for a successful planning of complex tumor resections in children. The efficiency of these techniques should be further analyzed in series with higher numbers and differentiations of tumors. PMID- 15750931 TI - Portal venous gas and surgical outcome of neonatal necrotizing enterocolitis. AB - PURPOSE: The prognostic significance of portal venous gas (PVG) in neonatal necrotizing enterocolitis (NEC) for operative intervention (OP), neonatal complications, and mortality remains uncertain. The authors designed a long-term prospective study to describe the natural history of PVG related to these outcomes and to test the hypothesis that PVG does not mandate OP. METHODS: All infants admitted to a single center between October 1991 and February 2003 were evaluated weekly to identify all cases of NEC (defined as Bell stage II or higher). Demographic, radiological, surgical, and outcome data were abstracted prospectively. Radiographic studies were performed at the onset of illness and at subsequent 6- to 8-hour intervals or as clinically indicated. A single pediatric radiologist reviewed all radiographs. Values are expressed as mean +/- SD. Odds ratios and relative risk ratios are reported with 95% CIs. The level of significance was P < or = .05. RESULTS: After the exclusion of 24 infants with lethal diseases, major congenital or chromosomal anomalies, or recurrent episodes of NEC, 194 of 5891 infants developed NEC. The overall incidence of NEC was 3.7%. In 194 infants with NEC, the incidence of PVG was 33% (n = 64). Gestational age (30.8 +/- 4 vs 29.3 +/- 4.2 weeks; P = .02) but not birth weight (1609 +/- 761 vs 1434 +/- 810 g; P = NS) was greater in infants with PVG compared with infants without PVG (n = 130). Sixty-six (34%) infants with NEC underwent OP. Operative intervention occurred more frequently in infants with PVG compared with infants without PVG (OR, 2.5; CI, 1.37-4.76; P = .003)--only 48% of infants with PVG underwent OP. Among the variables, gestational age, severe NEC (Bell stage III), severe intramural gas (in all 4 abdominal quadrants), and the presence of PVG, severe NEC was most highly associated with OP (OR, 77.47; CI, 10.36-580.16; P < .0001). Bell stage III NEC was present in 98% of infants who underwent OP compared with 40% of infants without OP ( P < .0001). Of all infants with NEC, 37 (19%) died. Mortality was higher among infants who underwent OP (33% vs 12%; P < .0003). A multivariate regression model identified Bell stage III (OR, 3.74; CI, 1.20-11.62; P = .02), but neither PVG nor OP, to be significantly associated with mortality. Of interest is that survival in infants with PVG was greater (but not significantly so) than in infants without PVG in both OP (74% vs 59%) and non-OP (91% vs 87%) groups. Furthermore, 30 of 64 (47%) infants with PVG survived without OP, and of all 33 infants with PVG who did not undergo OP, 30 (91%) infants survived. CONCLUSIONS: Decision for OP should be based on the severity of NEC and not on the presence of PVG alone because nearly half of infants with PVG survive without OP. Overall, the presence of PVG does not increase the risk of mortality among infants with NEC. Severe NEC, but not OP, is associated with higher mortality. PMID- 15750932 TI - Proteasome gene upregulation: a possible mechanism for intestinal adaptation. AB - BACKGROUND/PURPOSE: The mechanisms that control intestinal adaptation remain unknown. To better understand the adaptive process, microarray technology was used to analyze gene expression in a rat model of intestinal adaptation. METHODS: Adult male Sprague-Dawley rats underwent either a massive small bowel resection (70%) with anastomosis or a sham operation with small bowel transection and reanastomosis. After 21 days, ileal mucosa RNA was extracted. Individual RNA samples (n = 5 per group) were labeled and hybridized to 10 separate RAE 230A rat GeneChips. The signal values were calculated and the 2 groups were compared using a t test with the multiple testing correction of Benjamini and Hochberg (false discovery rate of 10%). Probe sets were analyzed for overrepresented physiologic pathways using Expression Analysis Systematic Explorer (EASE). RESULTS: Of the 15,866 probe sets on the RAE 230A GeneChip, 5437 probe sets were unexpressed and excluded. Of the remaining 10,429 probe sets, several overrepresented pathways (EASE score <0.01 after Bonferroni correction) were identified. Further analysis revealed that 13 probe sets related to proteasome degradation (an enzyme complex implicated in the regulation of cell proliferation) were significantly upregulated in the intestinal adaptation group compared to the sham group. CONCLUSIONS: Proteasomes may play a critical role in regulating the proliferation of intestinal mucosa during intestinal adaptation. PMID- 15750933 TI - Laparoscopic seromuscular colonic biopsies: a surgeon's experience. AB - BACKGROUND: Colonic dysmotility is a recognised cause of chronic constipation in children. Colonic dysmotility is better analysed by examination of the colonic muscle than rectal biopsy, which does not examine the defective area and has a low yield. We explored the role of laparoscopic colonic muscle biopsies to investigate children with intractable constipation. The authors describe the technique, its application, and results from a large series. METHODS: A retrospective review was conducted of all patients undergoing laparoscopic seromuscular colonic biopsies (hepatic flexure, mid-transverse colon, splenic flexure, and sigmoid colon) by a single surgeon for the investigation of chronic constipation over a 10-year period. Patient records were reviewed to determine the perforation frequency and management, postoperative recovery time and the frequency of an immunohistochemical abnormality. RESULTS: One hundred ninety seven patients (118 boys) were investigated by laparoscopic biopsy during the period. The mean age was 8.0 +/- 4.0 years (range, 1.4-22.4). The patients took 28.7 +/- 13.6 hours (range, 8-120) to recover, with 37 (19%) having nausea and/or vomiting requiring antiemetics. Most patients (160/197, 81%) were discharged the following day. Six patients (3%) had a mucosal perforation recognised at operation (treated by an Endoloop) with no change in postoperative outcome. Two patients (1%) had an unrecognised mucosal perforation requiring laparoscopic reoperation and Endoloop closure (laparotomy/colostomy not required), with no further sequelae. Eight-six patients (44%) had a specific immunohistochemical neuropeptide anomaly (reduced substance P [84], reduced vasoactive intestinal peptide [2]). CONCLUSIONS: Laparoscopic biopsy is a valuable tool to investigate chronic constipation in children, allowing a pathological diagnosis to be made in many cases. The complications of the procedure are acceptably low with this technique. PMID- 15750934 TI - The versatility of the umbilical incision in the management of Hirschsprung's disease. AB - BACKGROUND/PURPOSE: The aim of this report is to describe how the umbilical incision provides a quick and safe alternative to laparoscopy or other abdominal incisions in the management of Hirschsprung's disease (HD). METHODS: An analysis of 24 patients with HD, who were treated using an umbilical incision as part of their operative management between 1999 and 2003, was performed. RESULTS: There were 18 boys and 6 girls (mean age at diagnosis: 16.5 +/- 20.9 days). Twenty-one had rectosigmoid HD, and 3 had total colonic HD. Eighteen patients received a 1 stage transanal pull-through with transumbilical colonic biopsies at a mean age of 33.9 +/- 25.3 days, and a mean weight of 3.8 +/- 1.0 kg. Three patients with rectosigmoid disease had more complicated HD: 2 had a colostomy (1 enterocolitis, 1 extensive colonic dilatation), and 1 had an ileostomy (for perforated cecum). All subsequently underwent transanal pull-through. Three patients with total colonic HD had an ileostomy. All stomas were situated at the umbilicus. One of these patients subsequently underwent a Duhamel procedure via the umbilicus; 1 is still waiting; and 1 died of sepsis. Complications in this cohort included a seromuscular tear of the distal sigmoid at the level of the umbilical incision (1), infection at the umbilical incision requiring antibiotics (2), obstruction (1) and death (1 sepsis, 1 from Ondine's curse). The median time to start full feeds was 24 hours. For postoperative analgesia, 13 patients required acetaminophen only, and 9 patients received low-dose morphine. After a mean follow-up of 7.3 +/- 9.7 months, there has been an excellent functional and cosmetic result in the 22 survivors. CONCLUSIONS: The umbilical incision has all the benefits of a minimal access approach, but is less expensive and requires no specialized equipment or skill. The umbilical incision is an excellent, safe, and versatile alternative to laparoscopy for the treatment of patients with the full spectrum of HD. PMID- 15750935 TI - Colonic atresia without mesenteric vascular occlusion. The role of the fibroblast growth factor 10 signaling pathway. AB - BACKGROUND/PURPOSE: Colonic atresia occurs in 1:20,000 live births, offering a neonatal surgical challenge. Prenatal expression of fibroblast growth factor 10 (Fgf10), acting through fibroblast growth factor receptor 2b (Fgfr2b), is critical to the normal development of the colon. Invalidation of the Fgf10 pathway results in colonic atresia, inherited in an autosomal recessive pattern. Classically, disturbance of the mesenteric vasculature has been thought to cause many forms of intestinal atresia. The purpose of this study was to evaluate the role of vascular occlusion in the pathogenesis of colonic atresia. METHODS: Wild type (Wt), Fgf10(-/-), and Fgfr2b(-/-) mutant mouse embryos were harvested from timed pregnant mothers. Immediately following harvest, filtered India ink was infused via intracardiac microinjection. The gastrointestinal tract was dissected, and photomicrographs of the mesenteric arterial anatomy were taken at key developmental time points. RESULTS: Photomicrographs after India ink microinjections demonstrate normal, patent mesenteric cascades to the atretic colon at the time points corresponding to the failure of colonic development in the Fgf10(-/-) and Fgfr2b(-/-) mutants. The mesenteric arterial anatomy of the colon demonstrates no difference between the Wt and mutant colonic atresia. CONCLUSIONS: The absence of embryonic expression of Fgf10 or its receptor Fgfr2b results in colonic atresia in mice. India ink microinjection is a direct measure of mesenteric arterial patency. Colonic atresia in the Fgf10(-/-) and Fgfr2b(-/-) mutants occurs despite normal mesenteric vascular development. Thus the atresia is not the result of a mesenteric vascular occlusion. The patent colonic mesentery of the Fgf10(-/-) and Fgfr2b(-/-) mutants challenges an accepted pathogenesis of intestinal atresia. Although colonic atresia can occur as a result of vascular occlusion, new evidence exists to suggest that a genetic mechanism may play a role in the pathogenesis of this disease. PMID- 15750936 TI - Aberrations of the intrinsic innervation of the anorectum in fetal rats with anorectal malformations. AB - BACKGROUND: Fecal accumulation, constipation, soiling, and incontinence are common sequelae after repair of anorectal malformations (ARMs) in children. It is believed that besides the abnormalities of sacral roots, certain inherent abnormalities of the myenteric plexuses may play an important role in the final outcome after definitive repair. METHODS: This study was conducted to investigate the distribution of neuron-specific enolase (NSE), vasoactive intestinal peptide (VIP), and substance P (SP)-100 neurotransmitters in the rectosigmoid and fistulous tract of the ethylenethiourea-treated rat with ARMs. RESULTS: ARMs were induced by administering 1% ethylenethiourea (125 mg/kg) on gestational day 10, and the litter was harvested on gestational day 21 by cesarean section. Forty eight controls and 63 with ARMs (46 high-type and 17 low-type) were recovered. Whole-mount preparations of each rectosigmoid and fistulous communication between the rectum and genitourinary tract were stained with fluorescent antibodies against NSE, VIP, and SP-100. The tissues were counterstained with Eriochrome black-T and methyl green dyes to improve the visualization of the myenteric plexuses. CONCLUSIONS: The immunoreactivity of NSE, VIP, and SP-100 was markedly reduced in the rectum and fistulous tract of high-type ARMs and slightly reduced in low-type ARMs compared with controls. Intramural nerves stained by VIP and SP 100 antisera were decreased in both types of ARM, indicating that both inhibitory and excitatory motor neural elements were affected, and this may explain the distal colonic dysmotility seen postoperatively in both high and low ARMs. PMID- 15750937 TI - Notable sequential alterations in notochord volume during development in the Adriamycin rat model. AB - BACKGROUND/PURPOSE: The Adriamycin rat model (ARM) is a well-established model of the vertebral, anorectal, cardiac, tracheoesophageal, renal, and limb association. An important finding in the ARM is that Adriamycin induces abnormal notochord morphology in the region of the foregut. Having recently demonstrated notochord hypertrophy in ARM embryos, the authors designed this study to assess notochord volume sequentially from gestational days 10 to 14 (E10-E14) to test the hypothesis that notochord hypertrophy occurs maximally soon after Adriamycin administration. METHODS: Adriamycin (1.75 mg/kg) was administered intraperitoneally to pregnant rats on E7, E8, and E9. Control animals were given saline. Embryos were recovered at E10, E11, E12, E13, and E14 and embedded in paraffin. Quantitative morphology using the Cavalieri technique was performed on hematoxylin and eosin-stained transverse serial sections to determine total embryo and total notochord volume. RESULTS: The percentage volume of notochord per embryo was significantly increased (P < .05) in Adriamycin-treated embryos at all gestational time frames from E10 to E14 when compared with equivalent controls. This increased volume of notochord was found to be maximal at E11. CONCLUSIONS: These data support the authors' previous finding that Adriamycin induces notochord hypertrophy and suggest that notochord volume is increased relative to embryo volume soon after Adriamycin administration and is maximal on E11. The abnormal increase in notochord volume during the critical phase of development may interfere with organogenesis, resulting in the vertebral, anorectal, cardiac, tracheoesophageal, renal, and limb association. PMID- 15750938 TI - Bladder stone formation after sigmoidocolocystoplasty: statistical analysis of risk factors. AB - PURPOSE: Bladder stone formation (BSF) after sigmoidocolocystoplasty (SCP) is a major source of morbidity. The authors identify risk factors and analyze them statistically. METHODS: Medical records of 106 patients who had SCP at our institute were reviewed to examine the incidence of post-SCP BSF, the stone removal method and length of follow-up. The primary underlying pathological conditions were myelomeningocele in 103 patients and other conditions in 3. Risk factors for BSF were statistically assessed by comparing cases with and without BSF. RESULTS: Bladder stone formation occurred in 20 (18.9%) of 106 patients (BSF group). The remaining 86 did not have stones (NBSF group). Mean age at SCP was 11.7 years in the BSF group and 10.2 years in the NBSF group. Mean follow-up was 12.0 years in the BSF group and 11.0 years in the NBSF group. BSF developed after a mean of 30.3 +/- 25.6 months after SCP. Twenty-two stone removal procedures were performed in the BSF group and spontaneous evacuation occurred in 4 instances. Once stone free, BSF recurred in 4 patients. Although differences between the 2 groups with respect to sex, age at SCP, follow-up period, bladder capacity, bladder compliance, incidence of gram-negative bacteriuria, incidence of gram-positive coccus bacteriuria, electrolytes, frequency of urinary catheterization, post-SCP incidence of vesicoureteric reflux, and incidence of ureteric reimplantation were not statistically significant, differences in the incidence of spontaneous micturition, bladder neck tightening procedures, or gram positive bacillus bacteriuria, or frequency of bladder irrigation were all statistically significant (P < .05 in all, Fisher's Exact test). CONCLUSIONS: The authors are the first to statistically analyze a wide range of varying risk factors for BSF. BSF would appear to be common after SCP. PMID- 15750939 TI - Enhancement of intravascular sclerotherapy by tissue engineering: short-term results. AB - BACKGROUND/PURPOSE: Treatment of vascular malformations with sclerotherapy is often complicated by reexpansion secondary to endothelial recanalization. This study examined the use of an autologous fibroblast construct to enhance intraluminal scar formation after sclerotherapy. METHODS: New Zealand rabbits (n = 15) underwent ethanol sclerotherapy of a segment of the facial vein. After intraluminal saline flush, animals were equally divided into 3 groups. In group I, no further manipulations were performed. In groups II and III, collagen hydrogel was injected into the sclerosed vein, respectively, without and seeded with autologous green fluorescent protein-labeled fibroblasts. One week postoperatively, the vein segments were examined for patency and resected for histology. RESULTS: The sclerosed vein segments remained occluded in all animals. Histological examination of luminal thrombi demonstrated numerous viable fibroblasts in group III, whereas there were none in the control specimens from groups I and II. The presence of the injected autologous green fluorescent protein-labeled fibroblasts within thrombi of group III was confirmed by immunohistochemistry. CONCLUSIONS: An injectable tissue-engineered construct enhances sclerotherapy of the jugular vein in a leporine model by reliably delivering fibroblasts that populate the resultant thrombus. Further analysis of this novel therapeutic concept as a means to augment permanent scar formation and reduce luminal recanalization is warranted. PMID- 15750940 TI - Totally minimally invasive robot-assisted unstented pyeloplasty using the Zeus Microwrist Surgical System: an animal study. AB - BACKGROUND: Minimally invasive pyeloplasty is a difficult procedure even for an expert laparoscopic surgeon. The major difficulty is associated with the limitations of intracorporeal suturing and knot tying. Surgical robots, which hold minimally invasive surgical instruments, have wrists and provide tremor filtration and motion scaling that might be expected to facilitate complex procedures in newborns. METHODS: Seven survival piglets (4.0-7.5 kg) underwent a totally minimally invasive robot-assisted unstented pyeloplasty employing the Zeus Robotic Microwrist System. The ureter was transected at the level of the ureteropelvic junction and 8 mm was resected. The unstented anastomosis was fashioned with running suture and intracorporeal knot tying. The animals were recovered and intravenous urography was performed at 1 month. After sacrifice, the anastomosis and the kidney were evaluated grossly and histologically for leak, caliber, and healing. RESULTS: All animals survived the procedure without postoperative complications. The mean robotic setup time was 19 minutes (range, 10-30 min), mean anastomosis time 51 minutes (range, 39-63 min), and mean total operation time 76 minutes (range, 57-87 min). The urography showed hydronephrosis in the first animal. The other 6 animals had no abnormalities. Histopathology demonstrated severe hydronephrosis in the first pig and moderate hydronephrosis in the sixth and seventh. All other animals had no sign of hydronephrosis. All anastomoses were well healed and intact. CONCLUSIONS: Robot-assisted laparoscopic pyeloplasty is a technically feasible procedure with acceptable morbidity in an animal model. The robotic technology enhances surgical dexterity and precision. Robotic assistance can increase the applicability of minimally invasive surgery to complex procedures in children. PMID- 15750942 TI - The timing of delivery of infants with gastroschisis influences outcome. AB - BACKGROUND: The delayed onset of intestinal function in children with gastroschisis may be because of the injurious effects of amniotic fluid on the exposed bowel. This has led to consideration of early delivery to minimize intestinal damage and improve outcome, although this has not been carefully evaluated. The authors hypothesized that timing of delivery influences outcome in children with gastroschisis, and sought to evaluate the relative impact of factors that predict outcome in this disease. METHODS: All consecutive patients with gastroschisis (1992-2002) were divided into those delivered before ("early") or after ("late") 36 weeks. Bowel peel was described as "thin" or "thick," based on operative reports. Individual measures were analyzed by univariate analyses (chi2 /Student's t test), and logistic regression was used to identify significant factors for the length of stay (LOS) longer than the population average of 55 days. RESULTS: In 75 patients, 53.4% were "early" and 46.6% were "late." Groups were similar with respect to maternal age, birth weight, delivery mode, sex, and associated anomalies. Thickness of bowel peel was not affected by delivery time, yet "early" patients had significantly longer LOS and time to enteral feeds. Significant predictors of LOS more than 55 days included gestational age of 36 weeks or younger, time to enteral feeds of more than 26 days, and associated anomalies. Nonsignificant predictors included size of the defect, thickness of bowel peel, and need for silo. CONCLUSIONS: Delivery before 36 weeks is associated with longer hospitalization and increased tune to attainment of full feeds compared with later delivery. Fetal well-being should thus be the primary determinant of delivery for gastroschisis, as opposed to considerations regarding possible injurious effects to the bowel of prolonged gestation. PMID- 15750943 TI - Long-term effects of intrauterine exposure to mono-n-butyl phthalate on the reproductive function of postnatal rats. AB - PURPOSE: Although it is well known that phthalate esters induce testicular dysfunction in both adult and immature rats, there have been few reports on the long-term effect of phthalate esters on the testicular function of male rats exposed to phthalate esters in utero. This study was designed to assess the long term effects of the mono-n-butyl phthalate (MBP) ester on the testicular function of neonatal and adult rat offspring from pregnant dams exposed to phthalate esters during gestation. METHODS: Pregnant rats were administered MBP [0.5 g/(kg body weight/.d); 4 days] by gavage from the 15th to the 18th gestational day. Rats administered solvent only were used as control subjects. After the rats' puberty, using male pups whose testes descended normally, the authors examined their fertility while also measuring their testicular weights, mean seminiferous tubular diameter, and the developmental grade of the germ cells (Johnsen score) in their testes. Next, in neonatal rats, the authors measured the testicular concentration of the Mullerian inhibiting substance (MIS) protein using enzyme linked immunoassay and the expression level of the MIS messenger RNA using the quantitative polymerase chain reaction method as a marker of the Sertoli cells' function. Next the concentration of testosterone protein using a radioimmunoassay as a marker of the Leydig cells' function was measured. RESULTS: The pregnancy rate of the female rats coupled with MBP-treated male rats decreased significantly in comparison with that of the female rats coupled with control male rats (P < .01). Both the testicular weight and the Johnsen score in the MBP treated group were decreased significantly more than those of the control group (P < .05). Neither the concentration of the MIS protein nor the expression level of the MIS messenger RNA in the MBP-treated neonatal testes differed from those of the control testes, whereas the concentration of testosterone protein in the neonate testes decreased significantly in the MBP-treated group in comparison with that of the control group (P < .01). CONCLUSIONS: A prenatal short-time exposure to MBP induces a long-term effect on postnatal rats and impairs reproductive function in male offspring probably by inhibiting the Leydig cells' rather than Sertoli cells' function in the fetal period. PMID- 15750944 TI - Gubernacular cell division in different rodent models of cryptorchidism supports indirect androgenic action via the genitofemoral nerve. AB - PURPOSE: The role of the "gubernaculum" in testicular descent remains controversial. Androgens are proposed to act indirectly by the genitofemoral nerve (GFN) releasing calcitonin gene-related peptide. The authors studied the effects of sensory nerve ablation and androgen blockade on mitosis in the gubernacular tip to determine whether androgens act directly or indirectly. METHODS: Five rat models were examined for bromodeoxyuridine (BUdR)-labeling: (i) Sprague-Dawley (SD) rats (controls), (ii) prenatal flutamide-treated rats (75 mg/kg to dams on D16-19 gestation), (iii) neonatal capsaicin-treated rats (50 mg/kg, subcutaneous on day 0), (iv) congenitally cryptorchid transcrotal (TS) rats, and (v) capsaicin-treated TS rats (50 mg/kg, subcutaneous on day 0). Newborn rats were collected at days 0, 2, 4, 6, 8, and 10 (age, n = 5/model, n = 30) and were injected intraperitoneally with 1 mg/kg BUdR, 2 hours before killing. Histological sections of gubernaculum were examined immunohistochemically for BUdR labeling. RESULTS: In SD (control) rats, DNA synthesis in the gubernacular tip was high at birth, reached a peak at day 2, and then decreased progressively until day 10. A similar pattern was observed in TS rats. However, quantitatively, the levels were significantly higher. In flutamide treated rats, DNA synthesis was suppressed until day 6, similar suppression was observed in capsaicin-treated SD, and TS rats until day 4. CONCLUSIONS: Flutamide, a competitive androgen receptor blocker, reduces gubernacular mitosis to basal levels until day 6, highlighting the importance of androgen receptor. Excess DNA accumulation in TS rats is consistent with the known excess of GFN fibers and calcitonin gene-related peptide in this mutant. Capsaicin-inhibited mitosis in both day 2 SD and TS rats suggests that the GFN mediates androgen action on early postnatal gubernacular DNA synthesis and growth. PMID- 15750945 TI - The effect of growth hormone supplementation on late nutritional independence in pediatric patients with short bowel syndrome. AB - PURPOSE: The use of growth hormone (GH) supplementation for intestinal adaptation among adult patients with short bowel syndrome (SBS) has provided mixed results. This report examines the effect of GH supplementation on SBS in pediatric patients. METHODS: Two girls with SBS from neonatal gastrointestinal catastrophes received exogenous GH at 0.3 mg/kg per week subcutaneously and concurrent glutamine supplementation, beginning at 6 and 6(1/2) years of age. Changes in growth (height and weight) and changes in enteral and parenteral energy requirements were evaluated. RESULTS: Treatment duration was 8 and 2.5 years, respectively. Patient weights increased from the 5th to the 41st percentile and from the 17th to the 23rd percentile, respectively. Height increased from the 1st to the 57th percentile in the former patient and increased from less than the 1st to the 17th percentile in the latter. Both patients are independent of parenteral nutrition and take enteral nutrition alone. Tolerance for enteral diets was significantly improved in each girl, with only 2 stools per day maintained in one patient. CONCLUSIONS: The data show that late exogenous treatment with GH and glutamine supplementation improved growth parameters in pediatric patients with SBS. Growth hormone and glutamine supplementation may be beneficial in promoting late intestinal adaptation in pediatric patients with SBS. These data also suggest that these adjuncts may be useful in the early phases of intestinal adaptation. PMID- 15750946 TI - Torsion of the vermiform appendix. PMID- 15750970 TI - [The hemodiafiltration with endogenous reinfusion reduces the erythroid progenitor inhibition by uremic serum]. AB - PURPOSE: Anemia in end-stage renal disease (ESRD) patients shows a lower proliferation of erythroid progenitor cells such as burst forming unit-erythroid (BFU-E) than in normal subjects. As on-line hemodiafiltration with endogenous reinfusion(HFR) is thought to have a better biocompatibility and a wide range of uremic toxin removal, we compared the effect of serum obtained pre- and post standard hemodialysis (HD) and HFR dialysis performed in four ESRD patients with proliferation in normal subject) (controls) bone marrow BFU-E. METHODS: Mononuclear fraction was obtained by Ficoll-Hypaque density centrifugation and studies were performed in three different conditions: standard culture, adding serum from controls, adding serum from ESRD patients pre- and post HD and HFR dialysis. BFU-E were counted after 14 days with an inverted microscope and expressed as average scores from two dishes. Standardization between experiments was checked with a control culture for each experimental culture. RESULTS: The BFU-E proliferation rate was clearly reduced by adding serum from ESRD patients either pre-HD or pre-HFR. However, while this inhibition was exacerbated by post HD serum, it showed a significant reduction with post-HFR serum. CONCLUSIONS: This effect could be due to the removal of uremic toxins or to a lower dialysis induced cytokine release, both mechanisms involved in erythropoiesis inhibition in ESRD. PMID- 15750972 TI - [Alterations of cardiac output during bicarbonate hemodialysis and its relationship with other hemodynamic parameters]. AB - PURPOSE: Time course of cardiac output (CO) and other hemodynamic parameters were measured during hemodialysis (HD). Our aims were to identify a characteristic CO profile and investigate the relationship with other hemodynamic parameters. PATIENTS AND METHODS: CO was measured with ultrasound dilution method in 45 chronic hemodynamically stable HD patients. Diabetics and patients with heart diseases were excluded. Ultrafiltration rate (UFR) was fixed at 649 +/- 244 mL/min. Pre/post statistical comparisons were performed for CO, cardiac index (IC), central blood volume (CBV) and total peripheral resistance (TPR). RESULTS: CO was pre 5.7 +/- 1.8 and post 4.5 +/- 1.4 L/min (p=0.001); IC was pre 3.2 +/- 0.9 and post 2.6 +/- 0.7 L/m2 (p=0.001); CBV was pre 1.28 +/- 0.39 and post 1.09 +/- 0.32 L (p=0.001). TPR increased from 18.7 +/- 5.6 to 22.7 +/- 6.1 mmHg/L/min (p=0.001). Maximal CO reduction rate was found at 60 min, thereafter it reduced progressively. Log(CO1) increased in a non-linear way with body weight gain and similarly it decreased during UFR. A negative correlation was found between log(TPR1) and log(CO1-QA). CO reduction was associated with UFR and not with age, dialysis duration, left ventricular hypertrophy, sex and hemoglobin (Hb) in a multiple regression model (r2 =0.31, p=0.05). Qa/CO1 was 0.16 +/- 0.12. CBV/CO increased from 0.23 +/- 0.06 to 0.25 +/- 0.07%. CONCLUSIONS: Progressive CO reduction and TPR increase appear to be the typical hemodynamic features of bicarbonate HD with a UFR of moderate degree. Volume overload and CO increase were related in a non-linear way. TPR1 was strongly correlated with CO1-Qa, suggesting that a large arterovenous shunt was associated with increased resistance. PMID- 15750971 TI - [Long-term use of Profiler in patients with dialysis intolerance: 8 months follow up]. AB - PURPOSE: A new method of profiled dialysis has been set up for many years in the Department of Nephrology, Dialysis and Renal Transplantation at the University of Bologna. This profiled dialysis is based on the use of a new kinetic mathematical model, in collaboration with the Faculty of Engineering at the University of Bologna, for the elaboration of individual sodium and ultrafiltration profiles. OBJECTIVE: The profiled dialysis aims are: 1) to stabilize the intradialytic blood volume, boosting the refilling of plasma water from the intracellular and the extravascular to the extracellular/intravascular compartments, to balance the ultrafiltration; 2) to counteract the disequilibrium syndrome reducing the shift of water from the extra to the intracellular compartment. The pre-dialysis elaboration of profiles is completely automatic and supported by a computerized programme, Profiler, which has been included in the software of the dialysis machine Bellco Formula 2000 Plus. METHODS: In this prospective and multicenter study, this profiled dialysis, performed according to the Profiler, was continuously applied, for an 8-month period, in a group of 13 hemodialysis (HD) patients with an intolerance to previous dialysis treatment. During the study, the following parameters were evaluated, comparatively, with the patient's basal treatment: a) sodium and water balance; b) percentage incidence of intradialytic complications such as hypotensive events, cramps, headache, and vomiting and; c) metabolic and nutritional status. RESULTS: Results evaluated in comparison with the patient's previous dialysis treatments, demonstrated: a) plasma sodium from 136.8 +/- 3 to 136.8 +/- 1.7 mEq/L (p=ns), dry body weight from 72.2 +/- 19.3 to 71.7 +/- 19.5 kg (p=ns), heart index from 3.7 +/- 0.7 to 3.1 +/- 0.5 L/min/m2 (p=ns), reactance from 5.3 +/- 15 to 4.9 +/- 11 ohm (p<0.05); b) incidence of intradialytic hypotensive events reduced from 64 to 4% (p<0.001), cramps reduced from 8 to 1% (p<0.01); c) plasma albumin from 3.5 +/- 0.2 to 3.7 +/- 0.3 g/dL (p=ns), Kt/Veq from 1.3 +/- 0.1 to 1.36 +/- 0.2 (p=ns). CONCLUSIONS: Patients treated with profiled dialysis had a higher stability of intradialytic blood pressure (BP) achieving a reduction in the incidence of disequilibrium syndrome symptoms, in comparison with previous treatment. These clinical intradialytic improvements were not correlated to clinical, instrumental or biochemical indexes of sodium-water overload nor to a worst dialysis adequacy and nutritional state. PMID- 15750973 TI - [Long-term clinical experience in post-dilution HFR]. AB - PURPOSE: Hemodiafiltration reinfusion (HFR) is characterized by the use of regenerated ultrafiltrate as replacement fluid. We devised a new technique, post dilution HFR, aimed at increasing the purification efficiency, treatment tolerability and at reducing inflammatory state. METHODS: We performed post dilution HFR in six uremic patients during 18 months. Dialytic efficacy, filter blood rest and cytokine behavior were evaluated. RESULTS: Neither pyrogenic reactions nor other adverse phenomena were recorded. The tolerance to the treatment was excellent. We observed a high rate of urea extraction and optimal Kt/V values, a high extraction of beta2 microglobulin (beta2-m) and a reduction in blood rest; in addition, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) significantly decreased. CONCLUSIONS. The inversion of the standard HFR configuration allowed us to improve the removal of both urea and beta2-m, and the blood rest, with an optimal tolerability. Moreover, the reduction in cytokine levels could attenuate the uremic microinflammatory state. PMID- 15750974 TI - [Safety controls in a new hemodiafiltration on line technique (PHF)]. AB - PURPOSE: On-line hemodiafiltration (HDF) is gaining popularity due to increasing evidence of clinical benefits. The purpose of this study was to test a new on line technique paired hemodiafiltration (PHF). In addition, we evaluated the PHF system during in vitro contamination. METHODS: Five patients used the PHF technique over a 6-month period. We performed a disinfection protocol and tested for bacteria, endotoxin, halogenated carbons and metals in the feed water, and we tested for bacteria, endotoxins and fungi in the dialysate after different ultrafiltration stages. In vitro tests were performed using three bacterial concentrations of pseudomonas aeruginosa. Samples were analyzed from different sites throughout the entire on-line HDF circuit for bacteria endotoxins, fungus and the ability to stimulate whole blood production of tumor necrosis factor alpha (TNF-alpha). RESULTS: The bacteriological control from the feeding machine water and at the entrance to the monitors had a bacterial level of <100 CFU/mL. No bacteria were detected in the dialysate and endotoxin levels were <0.03 EU/mL. In the in vitro contamination study, with the three bacterial concentrations tested at various points in the circuit, bacterial and fungi were below the level of detection and endotoxins were <0.03 UE/mL. The addition of dialysate samples taken after the 1st microfiltration stage, as well as after the 1st and 2nd ultrafiltration stage and incubated with whole blood were not associated with stimulated TNF-alpha production. CONCLUSIONS: PHF appeared to be a safe and feasible method for on-line HDF even in the unforeseen presence of the bacterial contamination of the feed water or in the water distribution system. PMID- 15750975 TI - [Diffusive and convective treatments in the clinical practice: observation study of the Pavia district nephrology group]. AB - PURPOSE. Although convective treatments are widely used, there is no evidence that they can improve patient survival or hospitalizations nor clinical criteria indicating which patients could benefit from them. This study was carried out to evaluate the dialysis modality distribution and the clinical criteria used in choosing the dialysis therapy by the four dialysis Centers in the district of Pavia, a district characterized by a quite homogeneous population. METHODS: We evaluated age, gender, body mass index (BMI), time on dialysis, number of sessions per week, dialysis duration, dialysis modality, criteria used in choosing dialysis therapy, vascular access (VA), Kt/V, and the number of hypotensive episodes. RESULTS: Two hundred and seventy-two patients were enrolled in the study. Mean age was 67.3 +/- 12 yrs, BMI was 26.3 +/- 6.1, dialytic age was 5.49 +/- 5.5 yrs, Kt/V was 1.4 +/- 0.3, mean session time was 238 min. Fifty two patients (19.2%) were on a convective treatment. Age, BMI, time on dialysis, dialysis duration and number of sessions per week were no different between convective treatment patients and diffusive treatment patients. Kt/V was significantly different between convective and diffusive methods (1.55 +/- 0.37 vs 1.4 +/- 0.28, p<0.05). Convective treatments were prescribed for cardiovascular (CV) instability by the nephrologists from the Pavia district in the majority of patients (90.9%). There was no difference in hypotensive episodes between convective and diffusive methods. CONCLUSIONS: Our study demonstrated that lacking clinical evidence indicating an improvement in the long-term outcome with convective techniques, nephrologists in the Pavia district choose this type of dialysis treatment to ameliorate CV stability in uremic patients. PMID- 15750976 TI - [Clinical evaluation of the Paired Filtration Dialysis in children]. AB - PURPOSE: An important challenge for a pediatric nephrologist is to achieve in children the innovations of current adult dialytic technology. METHODS: Five children, mean age 14.1 +/- 1.5 yrs, mean weight 31.5 +/- 5.8 kg, mean dialytic age 32 +/- 20 months, were evaluated after a 6-month treatment with paired filtration dialysis (PFD). We used two capillary membranes: a 0.4 m 2 polysulfone hemofilter and a 0.8 m 2 cuprophan dialyzer. There was a reinfusion system (1540 +/- 150 mL/h) between these two filters. QD 500 mL/min and QB 230 +/- 10 mL/min. Ultrafiltration (UF) 2400 mL/h was planned for the hemofilter. At base-line and after 6 months of treatment, in addition, we evaluated routine clinical parameters, and other parameters such as beta2-microglobulin, dialytic adequacy (Kt/V) and nutritional status (dietetic diary). The data were analyzed using the Student's t-test for paired values. RESULTS: Medium values of small molecules did not demonstrate meaningful variations after 6 months of treatment. The extraction percentage after each session was between 63% phosphorous and 76% for urea, but only 25% for convection. Instantaneous urea clearance after 60 min was 178 +/- 10 mL/min. Dialytic efficiency and nutritional intake were appropriated: Kt/V 1.66 +/- 0.2; PCRn 1.65 +/- 0.2; protein intake 2.2 +/- 0.5 g/kg/die; caloric balance 71.2 +/- 15 Kcal/kg/die; protein balance 0.53 +/- 0.4 g/kg/die; and azotic balance 85.8 +/- 74 mg/kg/die. We confirmed the good depuration capacity from the middle molecules with convective treatments: beta2-microglobulin extraction was 53%. Moreover, in children we found good clinical tolerance to PFD with modest interdialytic symptomatology: headache (8%), hypotension (6%), and cramps and vomiting (<2%). Increased priming of the extracorporeal circuit was not a significant technical problem. The cuprophan membrane has been subsequently replaced with the polysulfone membrane. CONCLUSIONS: We can assert that this technique can also be performed in the pediatric age with similar results as in the adult age: good depuration of the small and averages molecules, good clinical tolerance, and shortening the dialytic sessions. PMID- 15750977 TI - [Aspects of the on-line hemodiafiltration with regeneration and reinfusion of the ultrafiltrate (HFR): multicenter study]. AB - Despite technological advances in dialysis treatment, survival, morbidity and the quality of life in hemodialysis (HD) patients are affected by long-term complications, often related to the treatment itself. Among these complications, moderate protein and caloric malnutrition are present in approximately 30% of dialysis patients and are viewed as major contributors to increased mortality. In malnutrition pathogenesis, great importance is given to protein catabolism and to the loss of somatic protein and amino acids during dialysis. On the contrary, toxin clearance is believed to influence, positively, both protein anabolism and dietary protein intake. In hemodiafiltration (HDF), the clearance process is potentiated by three mechanisms (diffusion, convection and adsorption) and this could have a favorable effect on malnutrition. In addition, the reinfusion of regenerated ultrafiltrate (UF) would avoid the loss of large amounts of useful solutes as occurs with standard HD. In fact, all amino acids are present in the UF, which is not important in standard HD, but could be a problem in hemodiafiltration reinfusion (HFR). We treated 16 patients with HFR during the previous 3 months (the study will last for 12 months). Patients had been previously treated with bicarbonate dialysis for at least 6 months. The clinical tolerance of HFR was excellent and the technique appeared to be quite simple. The preliminary biochemical results demonstrated the stabilization of some parameters (such as urea and uric acid) with an adequate clearance of small molecules, while variables related to nutritional status (body weight, serum albumin and serum transferrin) did not change substantially. Surprisingly, the loss of both branched chain amino acids (BCAA) and essential amino acids (EAA) seemed slightly lower in HFR compared with standard HD. However, the reduced loss of amino acids (AA) observed with HFR should take into account other factors, such as absorption on adsorbent material and the basal plasma AA concentrations. Therefore, although each patient is in control of himself, it is difficult to draw any definite conclusions after only 3 months. However, it is evident that the loss of AA in HFR is quite modest and is not increased by the fact that it is a hemofiltration technique with all the consequent positive effects. PMID- 15750978 TI - [A single centre study about the effects of HFR on anemia]. AB - PURPOSE: Erythropoietin (EPO) deficiency, erythropoiesis inhibition and reduction in red blood cell survival are the main causes of anemia in endstage renal disease (ESRD). Hemodiafiltration (HDF) with on-line endogenous reinfusion eliminates backfiltration and uses an ultrapure dialysate and reinfusate. This technique should improve anemia correction by increasing uremic toxin removal and reducing inflammatory cytokine production. In this study, we evaluated the effects of HDF with on-line endogenous reinfusion on anemia correction. METHODS: This was a single-center, prospective and non-randomized study. We selected 12 patients on EPO therapy with steady haemoglobin (Hb) values; eight patients were treated with bicarbonate dialysis and four patients with on-line HDF. We switched them to HDF with on-line endogenous reinfusion for 6 months, changing the EPO dose when Hb levels were not within the 11-12 g/dL range. Biochemical data were measured every month. Results are expressed as means +/- m.s.e. Data were analyzed using the Student's t-test and analysis of variance for repeated measurements. A value p<0.05 was considered statistically significant. RESULTS: Patients maintained the same Hb, ferritin and dialytic efficiency throughout the study. The EPO supplementation was significantly higher in patients treated with bicarbonate dialysis (108 +/- 8 UI/kg/week as compared with patients treated with on-line HDF (36 +/- 5 UI/kg/week) during the 6 months before treatment by HDF with on-line endogenous reinfusion. In the last 6 months, we detected a reduction in EPO consumption, but not statistically significant, in the patients switched from bicarbonate dialysis to HDF with on-line endogenous reinfusion (83 +/- 6 UI/kg/week). CONCLUSIONS: The change from bicarbonate dialysis to HDF with on line endogenous reinfusion caused a reduction in EPO supplementation and, consequently, a reduction in the cost of anemia therapy. It is necessary to increase the size of the study group and to prolong the observation period to possibly obtain statistical significances. PMID- 15750980 TI - [Bicarbonate balance in hemodiafiltration (HDF): a comparison between two infusion methods of on-line prepared solution]. AB - PURPOSE: Hemodiafiltration (HDF) has high removal rates of low and middle-high molecular weight uremic toxins. We aimed to understand the efficacy and the safety in correcting on-line HDF acidosis. We compared two infusion methods of on line prepared solution in HDF: HDF with an infusion solution produced from dialysate (HDF-OL) and HDF with a solution from patient ultrafiltrate after regeneration (HFR). METHODS: Eleven patients (four males, seven females) age 66 +/- 10 yrs, dialysis age 5.0 +/- 1.3 yrs, on anuria had two dialysis methods for the 1st session of the week, one HDF-OL and one HFR in 2 different weeks. In HDF OL a high-flux polysulphone dialyser 1.8 m2 was used, in HFR a two-stage filter was used: polyetersulfone 0.7 m2 + SMC 1.95 m2 and a sorbent cartridge Selecta plus (Bellco) to regenerate the ultrafiltrate. HCO3- in the dialysis bath was 32 mmol/L. RESULTS: Plasma bicarbonates, before dialysis were 21.6 +/- 2.1 mmol/L on HDF-OL and 21.5 +/- 3.3 on HFR (p=ns), at the end they were 27.5 +/- 1.8 mmol/L on HDF-OL and 27.8 +/- 1.2 mmol/L on HFR (p=ns). On HDF-OL bicarbonates reached a plateau at mid session: 27 +/- 1.2, 27.5 +/- 1.2, 27.5 +/- 1.8 to 120, 180 and 240 min respectively. On HFR the plateau was reached more slowly: 26.1 +/- 1.9, 27.1 +/- 1.4, 27.8 +/- 1.2 with the same times. CONCLUSIONS: HFR-OL and HFR efficaciously corrected acidosis in a 4-h dialysis session. The same results, statistically and clinically, were achieved with infusion solution derived from dialysate and from solution from regenerated ultrafiltrate. In the latter, it was interesting that the global quality of the infusion solution was obtained from a close circuit from the patient ultrafiltrate. PMID- 15750979 TI - [Comparison between two different on-line dialytic techniques: Standard HFR vs post-dilutional HFR]. AB - PURPOSE: Among hemodiafiltration (HFD) techniques, hemodiafiltration reinfusion (HFR) seems unable to achieve an optimal depurative efficacy. This study aimed to evaluate whether the HFR technique as suggested in its original configuration could be improved by devising a new technique (post-dilution HFR (PDHFR)) inverting the purification sequence to increase the purification efficiency of the entire system. METHODS: We performed standard HFR in six uremic patients during 6 months and, subsequently, during a further 6 months, PDHFR was performed. The dialytic efficacy of the two techniques and the filter blood rest were evaluated. In addition, we studied the behavior of cytokines during the inverted HFR sessions. RESULTS: We observed a significant increase in urea extraction and in Kt/V. An equally significant improvement was observed with regard to beta2-microglobulin (beta2-m) extraction and blood rest. Furthermore, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) significantly decreased after inverted HFR treatment. CONCLUSIONS: The inversion of the original configuration allowed us to improve the depurative efficacy of standard HFR, increasing the removal of both urea and beta2-m, and reducing the blood rest. Finally, it was notable that the reduction in cytokine levels could attenuate the uremic microinflammatory state. PMID- 15750981 TI - [On-line hemodiafiltration with endogenous reinfusion (HFR). Experience of a Centre]. AB - PURPOSE: Hemodiafiltration reinfusion (HFR) treatment is a dialysis technique that uses the endogenous reinfusion fluid and performs, simultaneously and separately, the three mechanisms of extracorporal depuration: diffusion, convection and adsorption. This study aimed to evaluate clinical and biochemical data of a group of six patients submitted to a dialytic HFR method for >6 months. METHODS: Six patients with a mean age of 53.8 +/- 11 yrs (five males, one female), treated with standard bicarbonate dialysis for a mean of 79.2 months, underwent HFR for a mean period of 14.9 +/- 6 months. Filters used were: a) in all patients polysulfone with 0.7 m2 of surface for the convection; b) polysulfone with 1.7 m2 in one patient, and modified cellulose with 2.0 m2 in five patients for diffusion; c) hydrophobic interaction resin and uncovered mineral carbon 240 mL for the adsorption. For all patients dialysis duration was 240 min and the amount of reinfusion fluid was 2.5 L/h as a mean, calculated according to blood flow and hematocrit (Hct), keeping a filtration fraction <22%. We evaluated, at different times, the following parameters: a) patient weight; b) Hct and erythropoietin (EPO) doses; c) parathyroid hormone (PTH); d) phosphatemia and doses of administered vitamin D; e) homocysteine (Hcy) and Beta2 microglobulin (Beta2-m); f) and albuminemia and transferrinemia as nutritional parameters. RESULTS: We observed an increase in Hct, with a reduction in EPO dosage, and an increase in albumin and transferrin levels, an improvement in nutritional indexes and in patient well-being. The mild increase in Hct with the same EPO dose was present in spite of a switch to intravenous (i.v.) administration from subcutaneous administration. There was low morbidity and only one hospitalization due to an infectious episode. CONCLUSIONS: HFR allows an amino acid saving and pro-inflammatory middle molecule removal, resulting in a better clinical situation for progressively critical uremic patients. PMID- 15750982 TI - [Acute biocompatibility of hemodiafiltration with endogenous reinfusion (HFR)]. AB - PURPOSE: In order to reduce the hemodialysis (HD)-induced pro-inflammatory activity we need to use a biocompatible dialysis membrane, avoid backfiltration and possibly use adsorbents. Hemodiafiltration reinfusion (HFR) is a new on-line hemodiafiltration (HDF) technique combining these aspects. This study aimed to evaluate the biocompatibility of the single dialysis session comparing standard HD and HFR. METHODS: Eighteen patients on chronic HD were enrolled in five Centers. Patients underwent one standard and two HFR study sessions; in each session we evaluated leukocyte activation at 0, 5, 15, 60 and 240 min; and interleukin-6 (IL-6), C-reactive protein (CRP) and IL-1 receptor antagonist (IL 1Ra) levels at 0, 60 and 240 min. RESULTS: Leukocyte activation was similar in HD and HFR, while the post-dialysis IL-6 increase was lower with HFR; CRP levels were stable during HFR, but increased after HD, and IL-1Ra did not demonstrate any difference. CONCLUSIONS: These preliminary data show that HFR still has a better biocompatibility in the single dialysis session. PMID- 15750983 TI - [Effects of different dialysis membranes and techniques on the nutritional status, morbidity and mortality of hemodialysis patients]. AB - To evaluate long-term effects of different hemodialysis (HD) membranes and techniques on nutritional status, morbidity, and mortality in HD patients, we prospectively studied 138 stable HD patients (59 females, 79 males, mean age 53 +/- 13 yrs) on maintenance HD from at least 1 yr with bicarbonate (BD) and cellulose acetate (AC). Patients were randomly assigned to one of four groups: comparable for age, sex, underlying nephropathy, time on dialysis, comorbidity, and followed-up for 5 yrs. Group A (n=38) BD/AC; group B (n=30) BD/low-flux polysulfone (PS); group C (n=30) BD/middle-flux PS or PA; group D (n=20) hemodiafiltration (HDF)/high-flux polysulfone (PS-HDF); group E (n=20) acetate free biofiltration (AFB) with PAN. Nutritional status was evaluated by anthropometric index, visceral protein compartment index, immunological index and bioelectrical impedance analysis. In all patients, we evaluated yearly plasma values of Beta2-microglobulin and of C-reactive protein (CRP) before and after dialysis. A significant and sustained improvement in nutritional status and a striking reduction in CRP and in pre- and post-dialysis beta2-microglobulin levels was observed in groups C, D and E. Morbidity (calculated from the number of clinical complications/patient/yr and from the number of hospital admissions/patient/yr) and mortality were significantly higher in groups A and B (mortality: group A=24%, group B=23%, group C=10%, group D=5%, and group E=5%; p<0.001). The results of our study indicate that the use of middle and high-flux biocompatible membranes is associated with a remarkable and sustained amelioration in nutritional status and with a significant improvement in the prognosis of HD patients, explained by the notable reduction in beta2 microglobulin and in the systemic inflammatory response. PMID- 15750984 TI - [Cystatin C, beta2-microglobulin and C-reactive protein in hemodiafiltration and on-line endogenous liquid reinfusion and in low flux polysulphone bicarbonate conventional hemodialysis]. AB - PURPOSE: Dialysis morbidity results partly from middle and large molecule retention, which is poorly removed by conventional hemodialysis (HD). The potential benefit of convective treatments could be the enhanced toxin removal over a wide molecular weight spectrum. This study aimed to evaluate cystatin C (cis), beta2-microglobulin (beta2-m) and C-reactive protein (CRP) removal behavior during hemodiafiltration reinfusion vs conventional low-flux HD (1.8 m2 low-flux polysulphone) (bicarbonate dialysis (BD)). The molecular weights of the substances evaluated in this study were as follows: cis = 13,300 daltons, beta2-m = 11,818 daltons, CRP = 160,000 daltons. METHODS: Twelve patients on stable HD (six males, six females), were enrolled; six patients underwent BD and six patients underwent HFR. We measured arteriovenous serum cis, beta2-m and CRP levels, in three consecutive mid-week sessions at the following periods: pre/post dialysis and after 60 min from the beginning of the session. At 60, 120 and 180 min of HFR, we collected the ultrafiltrate for cis, beta2-m, and CRP evaluation. RESULTS: Cis, beta2-m and CRP mean values did not differ at pre-dialysis in the two groups. Pre/post- dialysis difference for cis in HFR vs BD was statistically significant (p=0.002) because cis reduced in HFR and increased in BD during the session. Beta2-m and CRP pre/post- dialysis differences in HFR vs BD were not significant. Cis clearance, measured 60 min after the beginning of the session was 34.2 +/- 20.1 mL/min in HFR and 24.8 +/- 18.4 mL/min in BD (p<0.05). beta2-m and CRP clearances did not differ among the treatments. Regarding the ultrafitrate concentrations during the HFR session, cis significantly decreased (2.5 +/- 0.6 mg/dL at 60 min and 2.0 +/- 0.4 mg/dL at 180 min; p=0.004), as well as beta2-m (21.5 +/- 12.9 mg/dL and 19.0 +/- 14.1 mg/dL, respectively; p=0.02). Ultrafiltrate CRP values, as expected, did not differ during HFR. CONCLUSIONS: This study demonstrated that cis, a middle molecule, is well depurated in HFR, while in BD it increases. Beta2-m, although better removed in the convective phase during HFR, does not demonstrate a removal difference in HFR and in BD. CRP, a large molecule, does not have significant removal. Since cis and beta2-m have almost the same molecular weight, why do they have a different depuration? We need further studies to evaluate if membranes can remove these molecules or if protein electrical charges or their stereoscopy enables their removal. PMID- 15750985 TI - [C-reactive protein and homocysteinemia in patients undergoing on-line hemodiafiltration]. AB - PURPOSE: In renal disease therapy (RDT) patients, high plasma homocysteine (tHcy) is common and high C-reactive protein(CRP) levels can be observed, attributed to the inflammatory process caused by the dialysis itself. Hyperhomocysteinemia and bioincompatibility are considered independent vascular risk factors. This study evaluated the behavior of these parameters in patients undergoing on-line hemodiafiltration (OL-HDF). METHODS: In 56 patients, HDF was performed using high permeability polyamide membranes, exchanging in the post-dilution mode 16-18 L/session of a reinfusate obtained by the on-line system (triple filtration AK200, Gambro). CRP was measured by an immunological method at the start and the end of the session in patients without comorbidities (group 1, n=30)and with inflammatory diseases (group 2, n=26). In 23 of the 56 patients, tHCY was measured (by high performance liquid chromatography (HPLC)) before and after the mid-week session on different schedule of folinic acid, vitamin B12 and vitamin B6. RESULTS: Pre-dialytic CRP was in the normal range in group 1 patients, whereas it was higher in group 2 patients; dialysis did not induce a significant change in either group. The intradialytic percentage tHcy decrease was approximately 50% regardless of the pre-dialytic value, which was significantly different according to the vitamin supplements administered. CONCLUSIONS: HDF, as performed in this study, demonstrated biocompatibility and efficient Hcy removal; therefore, it can prevent cardiovascular disease (CVD) in patients on regular extracorporeal dialysis. PMID- 15750986 TI - [Can the treatment with L-carnitine improve the inflammation in chronic hemodialysis patients?]. AB - PURPOSE: Inflammation in patients on chronic hemodialysis (HD) is related to malnutrition and atherosclerosis; anemia is also often present in these patients. It has been demonstrated that l-carnitina treatment, in addition to reducing the need for erythropoietin (EPO), improves nutritional parameters and cardiac performance. METHODS: To evaluate the effect of l-carnitine on the inflammatory pathology in patients on chronic HD, we studied 11 patients with no sure signs of malnutrition, flogistic and infective pathologies and with C-reactive protein (CRP) <2 mg/dL. We evaluated at baseline, after 6 and 12 months CRP, serum albumin, hemoglobin (Hb),nPCR and EPO weekly requirement. RESULTS: We observed a reduction in CRP (from 0.88 +/- 0.65 to 0.42 +/- 0.17 mg/dL after 6 months and to 0.50 + 0.36 mg/dL after 12 months), an increase in serum albumin (from 10.9 +/- 1.23 to 2.08 +/- 1.88 and to 11.8 +/- 1.15 g/dL) and an increase in nPCR (from 0.96 +/- 0.09 to 1.15 +/- 0.2 and to 1.16 +/- 0.18 g/kg/die); EPO weekly requirement decreased (from 7363 +/- 2941 to 5909 +/- 3207 units after 6 months and to 5363 +/- 3139 units after 12 months). CONCLUSION: These results seem to underline a positive effect of l-carnitine on the inflammatory pathology of patients on chronic hemodialytic treatment. PMID- 15750987 TI - [On-line HFR and removal of uremic toxins inducing the loss of phospholipidic asymmetry of the erythrocyte membrane]. AB - The phospholipids of the erythrocyte membrane are normally distributed asymmetrically in the double layer with the aminophospholipid phosphatidylserine (PS) present only on the inside of the membrane, since its exposure on the outside has numerous physiopathological consequences. In previous studies we have observed that solutes retained in uremia cause increased exposure of PS on the outer surfaces of the erythrocyte membrane and that this phenomenon may be involved in the uremic physiopathology, reducing erythrocyte survival and encouraging abnormal erythrocyte-endothelium interactions. The capability of the extracorporeal blood clearance treatment in removing the circulating uremic factors, responsible for the increased exposure of PS in red blood cells (RBC), was evaluated in 6 chronic uremic patients treated with haemodialysis (HD) or with on-line HFR in a random cross-over perspective study. The PS removal was evaluated indirectly by measuring the expression of PS in normal RBC incubated with uremic plasma obtained at various moments of the clearance session. The capability of the uremic plasma to expose PS on the RBC of healthy subjects (n times increase compared to incubation of normal RBC with autologous plasma) was essentially unmodified during HD (3.3 +/- 0.2 pre HD; 3.3 +/- 0.1 after 2 hours; 3.1 +/- 0.2 at the end of the session) but was reduced during HFR (3.1 +/- 0.2 pre HD; 2.3 +/- 0.1 after 2 hours; 1.6 +/- 0.1 at the end of dialysis; p<0.001 at the end of dialysis vs pre and after 2 hours and p<0.001 vs HD at 2 hours and at the end of the session). The reduced capability of the uremic plasma obtained during the HFR session to expose PS in normal RBC, proves removal of the plasmatic uremic factors able to externalize the PS. To assess whether this removal effect is linked to the cartridge containing styrene resin used in the treatment with HFR, samples of ultrafiltrate were taken before and after the cartridge and its capability to express PS on normal RBC was measured. The absolute RBC values expressing PS (%) were (pre-cartridge vs post-cartridge) 8.6 +/- 0.3 vs 3.8 +/- 0.2 after 5 minutes from the start of the session; 3.9 +0.1 vs 1.6 +0.2 halfway through the session; 3.1 +/- 0.1 vs 1.3 +/- 0.66 at the end of the session (p<0.005 pre vs post at all times). Our results show that uremic compounds able to cause increased exposure of PS in RBC can be removed during on line HFR, mainly thanks to the adsorption properties of the cartridge containing resin. This removal might be of benefit to uremic patients, improving the anaemic condition and reducing abnormal RBC-endothelium interactions which may contribute to endothelial disorder during uremia. PMID- 15750988 TI - [Cardiovascular risk markers in hemodialysis]. AB - PURPOSE: Although maintenance dialysis prevents death from uremia, patient survival remains an important issue. Cardiovascular (CV) events have been considered the main cause of death in hemodialysis (HD) patients. Some authors demonstrated an expected remaining life span of < or =2 yrs in HD patients who had a myocardial infarction. Therefore, it is very important to appraise risk factors and to adopt the correct diagnostic approach to match therapy. Nevertheless, acute myocardial infarction can be misdiagnosed in uremic patients, because typical markers have high false positivity rates. It has been estimated, for example, that 29% of HD patients have elevated serum troponin T concentrations, but do not have evidence of myocardial injury. Troponin T is more frequently elevated than troponin I among asymptomatic patients with renal insufficiency and this could be due to the relatively higher levels of an unbound cytosolic pool of troponin T and its higher molecular weight. Neither the common cardiac markers (LDH, LDH 1, CPK, CK-MB) are sensitive or specific as in the general population, but a recent 2-yr observational study showed that pre dialytic high serum concentrations of troponin T and CK-MB mass were associated with complete mortality, cardiac mortality, myocardial infarction and unstable angina (MACEs). In our study, we evaluated how dialysis influenced serum troponin I and CK-MB mass, and then we assessed serum homocysteine (Hcy), an additional CV risk factor in uremic patients. METHODS: We studied 17 uremic patients (13 males, four females) on standard HD and six patients (four males, two females) on on line hemodiafiltration (HDF), who were taking folic acid for at least 3 months. Patients who suffered from acute or chronic cardiac ischemic disease were excluded. We performed arterial gas-analysis, Na+, K+, Ca++, Mg++, Cl-, P, serum albumin, creatinine (Cr), urea, total homocysteine (tHcy), red blood count (RBC), troponin I and CK-MB mass, both pre and post-dialysis. We assessed urea reduction rate percentage (URR%), Kt/V, Hcy percentage reduction ratio (ORR%), and anthropometric parameters. RESULTS: Anthropometric parameters, pre- and post dialytic pH, HCO3 and electrolytes did not differ between the two groups, Kt/V and URR%. Even in on-line HDF, ORR% directly correlated with KtV and URR% (r=0.79, p<0.04; r=0.76, p<0.05, respectively). Troponin I and CK-MB mass were not significantly different in pre- vs post-dialysis, both on standard HD and on line HDF. Nevertheless, in standard HD, post-dialytic troponin I correlated with serum sodium concentration (r=0.93, p<0.000), potassium (r=0.67, p<0.004) and serum chlorine (r=0.92, p<0.92, p<0.000). CK-MB mass showed a correlation with serum chlorine (r=0.49, p<0.05). Post-dialytic CK-MB mass correlated with serum potassium for on-line HDF (r=0.83, p<0.03). CONCLUSIONS: Our study suggests the probability that dialytic adequacy improves CV outcome causing a reduction in the concentration of homocysteinemia and it demonstrates that convective treatments (on-line HDF) are best in reaching this end-point. Our data suggests that hemodialytic treatments, both standard HD and on-line HDF did not modify serum troponin I and CK-MB mass. We can use these parameters as a diagnostic approach in acute or chronic cardiac ischemic disease in HD patients, because they are not influenced by the hemodialytic procedure. This allows the selection of high risk patients, and offers them on-line treatment as the best suitable therapeutic option. PMID- 15750989 TI - [Vitamin B12 clearance (Kd-B12) in hemodialysis (HD) and hemodiafiltration (HDF)]. AB - PURPOSE: The dialysis dose is usually assessed by Kt/V urea; however, it is possible that middle molecule (MM) removal could play a role in optimal treatment. Vitamin B12 is a classical MM marker and Kd-B12 is used to compute a MM-based dialysis index, requiring a weekly total clearance (Kd-B12 + renal creatinine clearance (CrCl) > or =30 L, corresponding to IDB12> or =1 (Babb et al, Kidney Int, 1975). Recently, it was demonstrated that by increasing the total Kd-B12 per session (TCV) from 10 to 16 and to 26 L, the relative risk (RR) of death was reduced from 1 to 0.79 and to 0.62, respectively (Leypoldt et al, Am J Kidney Dis 1999). This implies that a minimum TCV of 16 L, but preferably of 26 L per session, should be delivered, for anuric HD patients on a 3x/wk schedule. To extend these results to the whole hemodialysis (HD) population, we suggest transforming TCV into the corresponding IDB12 values: i.e. a TCV=10 L on 3x/wk corresponds to IDB12=1, a TCV=16 and 26 L corresponds to IDB12=1.6 and 2.6, respectively. METHODS: This study aimed to assess Kd-B12 and IDB12 for all stable patients in our unit. There were 62 patients (33 males, 29 females): five patients were being dialyzed once per week (1x), nine patients twice (2x), 46 patients three (3x) and two patients four times (4x) per week (wk); the session length was 232+/-18 min. Most dialyzers had a large surface area (mean 1.9+/-0.3 m2), with KoA-B12=211+/-92 mL/min. Eleven patients, 3x/wk, were on hemodiafiltration (HDF): the reinfusion rate was 33+/-3 mL/min in five patients (sHDF) and 76+/-12 mL/min in six patients (HDF on-line (OL). Kd-B12 was computed as a function of KoA-B12, effective plasma flow, Qd and ultrafiltrate (UF). IDB12 was computed from Kd-B12, ses-sion length and schedule, CrCl and body surface area. RESULTS: The main results are given below: [table: see text] On average, Kd B12 was 105 +/- 13 mL/min on HD and 152+/-34 mL/min on HDF. A significant difference was found only for HDF-OL and was essentially due to the higher UF. Of note, the presence of renal function allowed good IDB12 values for 1x/wk and 2x/wk patients, even better than for the standard 3x/wk patients. CONCLUSIONS: We have demonstrated that most available dialyzers provide high Kd-B12 values (but HDF-OL performs significantly better) and that IDB12, by quantifying the impact of UF, session length, schedule and renal function, allows the assessment of dialysis adequacy beyond Kt/V urea, for all HD or HDF patients, on a routine basis and at no added cost. PMID- 15750991 TI - [Urea distribution volume and ionic dialysance]. AB - PURPOSE: Direct dialysis quantification (DDQ) represents the gold standard for determining urea distribution volume (V) in hemodialysis (HD) patients, but is impractical for routine use because it requires equilibrated post-dialysis plasma water urea concentration. The "formal" single-pool, variable volume, urea kinetic model (SPVV-UKM) is easier to use, needing a blood sample drawn immediately after the dialytic session, but to obtain a V value consistent with the DDQ method, it requires a correct estimate of dialyzer urea clearance (Kd), actually often overestimated. Ionic dialysance (ID) accurately estimates the "effective" urea clearance (Keff), namely Kd corrected for total recirculation. Using ID as an input parameter to SPVV-UKM, correct V values are expected when end-dialysis plasma water urea concentrations are determined in a blood sample drawn after the blood pump speed has been reduced to 50 ml/min for 2 min (Upwt2'). OBJECTIVE: We aimed to compare the V values determined by SPVV-UKM, ID and Upwt2' (VID), those determined by "formal" SPVV-UKM (VKd) and those determined by the anthropometric method proposed by Watson (VA), with the V values determined by the gold standard DDQ method (VDDQ). METHODS: Thirty-one anuric patients on chronic thrice-weekly HD were studied in 31 dialysis sessions (one per patient). RESULTS: VDDQ = 26.5 +/- 5.3 L; VID = 26.2 +/- 5.1 L; VKd = 32.1 +/- 5.7 L;. VA = 33.2 +/- 5.8 L. The mean (VID - VDDQ) difference was -0.2 +/- 1.3 L, not statistically significant (95% confidence interval (95% CI) -0.7 to 0.2 L; p=0.302); the mean (VKd - VDDQ) difference was 5.6 +/- 2.3 L, statistically significant (95% CI 4.7 to 6.4 L; p<0.001); the mean (VA - VDDQ) difference was 6.7 +/- 2.7 L, statistically significant (95% CI 5.7 to 7.7 L; p<0.001). CONCLUSIONS: ID use as an input parameter to SPVV-UKM allows adequate V determinations and, at the same time, circumvents the problem of delayed post-dialysis blood samples. On the other hand, the use of "formal" SPVV-UKM or of anthropometric equations leads to a significant overestimation in urea distribution volume. PMID- 15750990 TI - [Hypertension and fluid volume increase in uremic patients treated with convective hemodialysis: electric bioimpedence analysis]. AB - PURPOSE: Hypertension is considered an important cardiovascular (CV) risk for uremic patients on hemodialysis (HD). Its main pathogenetic factor is the extracellular fluid volume increase. METHODS: This study aimed to evaluate the water distribution by electric bioimpedence (BIA) in hypertensive uremic patients treated with convective hemodialysis (HD) (group A) and with diffusive HD (group B). The patients were on antihypertensive therapy: angiotensin-converting enzyme (ACE)-inhibitors, beta blockers, calcium antagonists and angiotensin II (ATII) antagonists. RESULTS: The BIA analysis demonstrated that the total and extracellular body water had the same distribution in the two groups with no statistical differences (Student's t-test); although the interdialytic weight gain was higher in group B than in group A. CONCLUSIONS: Blood pressure (BP) was treated by less drug use in patients on convective HD than in patients on diffusive HD. PMID- 15750992 TI - [Physical exercise during hemodialysis session: effect on quality of life]. AB - There are many studies showing beneficial psychophysical effects of exercise in dialyzed patients. Moreover, it has been suggested that exercise positively correlates with better metabolism, better blood pressure control and with total hemoglobin. In our dialysis unit eight dialyzed patients (average age = 66.7 years), for eight weeks participated in physical training with bike (Reck Moto Med Letto) during dialysis treatment. Controls of glucose metabolism, blood pressure and dialysis efficiency index (Kt/V and URR) at rest and during exercise was performed. All patients responded well to exercise and expressed better muscular performance during and after exercise time. Our study showed in all patients improvement of Kt/V and URR index after physical exercise period, compared to exercise free time (p < 0.005). We suggest that exercise during dialysis treatment is safe and consents either better psychophysical performance or better dialytic efficiency. PMID- 15750993 TI - [Risk of arrhythmias in hemodialysis patients vs healthy people]. AB - PURPOSE: Hemodialysis (HD) patients present an elevated risk of ventricular arrhythmias and sudden death. This risk is correlated with QTc dispersion, assessed as the difference between maximum and minimum QT recorded on 12 leads. Our study aimed to estimate the difference between QT, QTc, QT dispersion and QTc in HD patients on standard HD and online hemodiafiltration (HDF) in pre- and post dialysis vs. normal controls. METHODS: Nineteen uremic patients on standard HD (13 males, six females) and nine patients (six males, three females) treated with on-line HDF were studied. Seven normal subjects were the control group, matched for age and sex. No one was taking drugs that could interfere with electrocardiographic morphology and QT value. Basal anthropometric parameters were taken and simultaneous 12-lead electrocardiograms (ECGs) were recorded before and 30 min after HD. Serum concentration of creatinine (Cr), urea and electrolytes (Na, K, Mg, Ca, Pi) were monitored before and 30 min after HD, in the short interval section. QT interval was measured according to the Bazett formula, in the presence of different heart rates: QTc=QT/RR(0.5) (heart rate corrected QT interval). RESULTS: Anthropometric parameters were similar among the groups. Before dialysis, standard HD patients had lower calcium and higher potassium than the controls (Student's t-test): (p<0.01, p<0.004). ECGs in the two groups did not show essential differences. After dialysis calcium was higher and magnesium lower in HD patients than in controls: (p<0.0006, p<0.000); QT and QTc dispersion was higher in HD patients than in controls: (p<0.01, p<0.04). Before dialysis on-line HDF patients had lower magnesium (p<0.01) than controls; while there were no electrocardiographic differences between them. In the post dialytic phase, calcium was higher and phosphates, magnesium and potassium were lower than in controls: (p<0.001, p<0.01, p<0.002, p<0.01); QT and QTc dispersion was higher in HD patients (p<0.03, ns). Controls did not demonstrate any correlation between electrolyte and electrocardiographic parameters; while QTc dispersion and phosphates positively and directly correlated in uremic standard HD patients after dialysis (r=0.48, p<0.02), and then QTc dispersion correlated negatively to Ca/P ratio (r=-0.63, p<0.003). PMID- 15750994 TI - [Ionic dialysance: which relationship with effective urea clearance?]. AB - PURPOSE: It has been suggested that ionic dialysance (ID) can adequately estimate the "effective" urea clearance (eK), i.e. urea clearance corrected for total (access and cardiopulmonary) recirculation. Unfortunately, the results obtained by different authors in determining the relationship existing in vivo between ID and eK do not always agree. Furthermore, it has been recently evidenced that ID values could be different, according to the different methods used to modify the inlet dialysate conductivity during ID measurement. OBJECTIVE: We aimed to verify the relationship between the mean values of repeated instantaneous ID determinations and the urea clearance values corrected for access recirculation, urea clearance corrected for total recirculation (eK) and urea clearance corrected for both total recirculation and post-dialysis urea rebound (body urea clearance), determined according to the direct quantification method (dKDDQ, eKDDQ and bK, respectively). METHODS: Thirty-one anuric patients on chronic thrice-weekly hemodialysis (HD) were studied in 31 dialysis sessions (one per patient), performed using Integra machines equipped with the Diascan module for the automatic ID determination and the Quantiscan module for the fractional collection of outlet dialysate. The mean values of repeated ID determinations at 30 min intervals throughout each dialytic session by the Diascan module were compared with dKDDQ, eKDDQ and bK values. RESULTS: ID = 179 +/- 24 mL/min; dKDDQ = 200 +/- 27 mL/min; eKDDQ = 188 +/- 26 mL/min; bK = 165 +/- 25 mL/min. The mean (ID - dKDDQ) difference was -21 +/- 10 mL/min (95% confidence interval (95% CI) 25 to -17 mL/min; p<0.001). The mean ID/dKDDQ ratio was 0.90 +/- 0.05, indicating a mean difference between dKDDQ and ID of 10%. The mean (ID - eKDDQ) difference was -9 +/- 9 mL/min (95% CI -12 to -6 mL/min; p<0.001). The mean ID/eKDDQ ratio was 0.96 +/- 0.05; therefore, indicating a mean difference between eKDDQ and ID of only 4%. The mean (ID - bK) difference was 15 +/- 7 mL/min (95% CI 13 to 17 mL/min; p<0.001). The mean ID/bK ratio was 1.09 +/- 0.05, indicating a mean difference between bK and ID of 9%. CONCLUSIONS: The mean value of repeated ID determinations could be considered clinically as an adequate estimate of urea clearance corrected for total recirculation (eKDDQ). Given that ID determination does not require either blood sampling or laboratory tests, therefore, it becomes possible to estimate easily and rapidly, once the urea distribution volume (V) has been correctly determined, the Kt/Vsp at each dialytic session. PMID- 15750995 TI - Cancer caregiving and subjective stress: a multi-site, multi-dimensional analysis. AB - Although research has emerged documenting the psychosocial impact of family care for cancer patients, few efforts capture the multi-dimensional nature of cancer caregiving stress, particularly among socioeconomically diverse samples. Utilizing data collected from cancer caregivers at a non-urban, Southern US site and an inner-city, Northeastern US site (N=233), the present study identified predictors of multiple dimensions of caregivers' subjective stress (i.e. emotional appraisals of care demands). Various indicators representing the sociodemographic context of care, cancer care demands, and psychosocial resources were found to exacerbate or buffer caregivers from feelings of exhaustion, role entrapment, and loss of intimacy with the cancer patient. The multivariate regression model also emphasized the diffuse yet potent role care recipient mood problems and caregiver mastery/optimism have on multiple dimensions of subjective stress. The findings offer a number of recommendations for future research and practice focused on informal cancer care. PMID- 15750996 TI - A novel mycolactone from a clinical isolate of Mycobacterium ulcerans provides evidence for additional toxin heterogeneity as a result of specific changes in the modular polyketide synthase. PMID- 15750997 TI - Does it hurt to know the worst?--psychological morbidity, information preferences and understanding of prognosis in patients with advanced cancer. AB - BACKGROUND AND METHOD: Many doctors, while offering open discussion of diagnosis and treatment to cancer patients, still express concern about the psychological impact of discussing a poor prognosis. In this study, 106 advanced cancer patients were interviewed at home. The interview included open discussion and structured questionnaires: the Hospital Anxiety Depression Scale and Rotterdam Symptom Checklist. Participants were asked about information preferences, worries and unanswered questions. The interviewer (M.B.) rated "Understanding of diagnosis and of prognosis". RESULTS: A majority of participants (78%) demonstrated a good understanding of their diagnosis and were consistent with their information preference. On prognosis, 54% of those desiring information (49% of overall group) were fully aware; a further 22% were aware but unrealistic about time-scale. Although most were satisfied with their level of knowledge, 28% had further questions, particularly concerning "mode of disease progression" (n=25), "mode of death" (n=13) and prognosis (n=18). When understanding of prognosis was compared with psychological distress, a significant difference was observed between those with realistic versus unrealistic time-scales. However, this could be explained by increasing physical symptoms. CONCLUSIONS: Patients with advanced cancer have a good understanding of their diagnosis but many do not fully understand their prognosis. Although patients with deteriorating health are more likely to be psychologically distressed, awareness of prognosis does not itself cause depression. Doctors should be aware of patients' ongoing but often unvoiced concerns and prepared to explore these with them. PMID- 15750998 TI - Electrostatic effects on the thermodynamics of protonation of reduced plastocyanin. AB - The L12E, L12K, Q88E, and Q88K variants of spinach plastocyanin have been electrochemically investigated. The effects of insertion of net charges near the metal site on the thermodynamics of protonation and detachment from the copper(I) ion of the His87 ligand have been evaluated. The mutation-induced changes in transition enthalpy cannot be explained by electrostatic considerations. The existence of enthalpy/entropy (H/S) compensation within the protein series indicates that solvent-reorganization effects control the differences in transition thermodynamics. Once these compensating contributions are factorized out, the resulting modest differences in transition enthalpies turn out to be those that can be expected on purely electrostatic grounds. Therefore, this work shows that the acid transition in cupredoxins involves a reorganization of the H bonding network within the hydration sphere of the molecule in the proximity of the metal center that dominates the observed transition thermodynamics and masks the differences that are due to protein-based effects. PMID- 15750999 TI - Lipidic membranes are potential "catalysts" in the ligand activity of the multifunctional pentapeptide neokyotorphin. AB - Neokyotorphin (NKT) is a multifunctional pentapeptide that is involved in biological functions as diverse as analgesia, antihibernatic regulation and proliferation stimulus of tumour cells. The interaction of neokyotorphin with cell membranes is potentially important to all these multiple biological processes since receptor-mediated processes are thought to be involved in neokyotorphin action. Sargent and Schwyzer proposed in their "membrane catalysis" model that ligands interact with membrane lipids in order to adopt the necessary conformation for cell receptors. We have used fluorescence techniques to study the depth, orientation and extent of incorporation of NKT with model membrane systems (lipidic vesicles). The roles of lipid charge, membrane phase and sterol presence were investigated. The phenolic ring of tyrosine is located in a shallow position in membranes. The extent of partition is less in gel crystalline membranes than in liquid crystalline membranes. Addition of cholesterol causes a reorientation of the tyrosine ring at the interface of lipidic bilayers. Lipidic membranes meet all the conditions required for acting as potential "catalysts" in the ligand activity of the multifunctional pentapeptide NKT, because they modulate the exposure and orientation of the phenolic ring, which is most likely involved in docking to receptors. PMID- 15751000 TI - Photocontrollable self-assembly. AB - The incorporation of photoswitching molecules into molecular building blocks creates the possibility of photoresponsive self-assemblies in which the self assembled architecture or self-assembling process can be controlled by external light stimulus. Among the photoswitching molecules, azobenzene has been used most widely by virtue of the large photoinduced changes in its molecular geometry and physical properties. This article reviews how azobenzene can be effectively used to construct the self-assemblies in which supramolecular structure and formation/dissociation can be altered by light. PMID- 15751002 TI - Quality of life and current coping in young adult survivors of childhood cancer: positive expectations about the further course of the disease were correlated with better quality of life. AB - OBJECTIVES: As a result of advances in the treatment of childhood cancer many patients who may previously have had a limited life expectancy, are now surviving into adulthood. More insight is needed into the long-term adjustment of young adult survivors of childhood cancer. The purpose of this study was to (1) assess health-related quality of life (HRQoL), and (2) to explore the role of cognitive coping in relation to HRQoL. METHODS: HRQoL of 353 Dutch young adult survivors of childhood cancer was compared with HRQoL of 507 peers. Linear regression analyses predicted survivors' HRQoL by cognitive coping, independent of the impact of demographics and medical variables. RESULTS: Survivors reported a lower HRQoL than their peers. Health status was the best predictor of the Physical Component Scale of the RAND-36; health status and cognitive coping contributed almost equally well to the Mental Component Scale. The explanatory value of cognitive coping could mainly be attributed to the use of predictive control strategies. CONCLUSIONS: Because current coping seemed to be an important predictor of HRQoL, interventions directed at the coping strategies of survivors should be useful. The strong association between predictive coping and HRQoL stresses the importance of focusing at having positive expectations about the further course of the disease. PMID- 15751001 TI - Synthesis and antiapoptotic activity of a novel analogue of the neutral sphingomyelinase inhibitor scyphostatin. AB - The enantioselective synthesis of an analogue of scyphostatin, a potent inhibitor of the neutral sphingomyelinase, is described. The synthesis starts with cyclohexanone and a protected D-serine derivative. The key step is an asymmetric hydroxylation to access a hydroxycyclohexanone, which is transformed into a substituted hydroxycyclohexenone. This is converted into the scyphostatin analogue 14, a chemically and metabolically stabilised compound lacking the epoxy function of the natural congener and carrying a palmitic acid group instead of the native trienoyl residue. An evaluation of the biological activity of 14 revealed neutral sphingomyelinase inhibition in several in vivo test systems (monocytes, macrophages, hepatocytes) monitoring antiapoptotic effects and the inversion of phorbolester-induced translocation of green fluorescent protein labelled kinase (protein kinase C-alpha). PMID- 15751003 TI - Proteomic changes in rat serum, polymorphonuclear and mononuclear leukocytes after chronic nicotine administration. AB - In order to gain information about the effect triggered at the molecular level by nicotine, its neuroimmunomodulatory properties and its impact on the pathogenesis of inflammatory diseases, peripheral blood serum and leukocytes of rat submitted to passive nicotine administration were subjected to proteomic investigation. Serum, polymorphonuclear (PMN) and mononuclear (MN) leukocytes from chronically treated animals and from control animals were analysed by a two-dimensional (2-D) gel electrophoresis/mass spectrometry approach to detect differentially expressed proteins. The nicotine regimen selected is known to have a stimulatory effect on locomotor activity and to produce a sensitisation of the mesolimbic dopamine system mechanism involved in addiction development. After 2-D gel analysis and matching, 36 spots in serum, seven in PMN and five in MN were found to display a statistical difference in their expression and were subjected to matrix-assisted laser desorption/ionization-time of flight-mass spectrometry peptide fingerprinting for protein identification. Fifteen different proteins were identified. The results indicate an overall impact of nicotine on proteins involved in a variety of cellular and metabolic pathways, including acute phase response (suggesting the effect on inflammatory cascades and more in general on the immune system), oxidative stress metabolism and assembly and regulation of cytoskeleton. In particular, the observed changes imply a general reduction in the inflammatory response with a concomitant increased unbalance of the oxidative stress metabolism in the periphery and point to a number of potential noninvasive markers for the central nervous system (CNS) and non-CNS mediated activities of nicotine. PMID- 15751004 TI - No effect of encapsulation on the pharmacokinetics of warfarin. AB - BACKGROUND: In double-blind comparator studies with the oral direct thrombin inhibitor (oral DTI) ximelagatran, warfarin (Coumadin) was administered in encapsulated form in order to maintain patient and investigator blinding. This open, randomized, two-way crossover study was conducted to determine whether the encapsulated warfarin tablets (Coumadin) used in the ximelagatran studies are bioequivalent to nonencapsulated, commercially available warfarin (Coumadin) tablets. METHODS AND RESULTS: Eighteen healthy men received two 2.5 mg tablets of encapsulated warfarin and two 2.5 mg tablets of nonencapsulated warfarin as single oral doses, 14 days apart. The 90% confidence intervals for the mean treatment ratio (encapsulated tablet/nonencapsulated tablet) fell within the limits considered to reflect bioequivalence (0.80, 1.25) for total area under the plasma concentration-versus-time curve (AUC(infinity)), AUC to the last evaluable concentration (AUC(t)), and maximum plasma concentration (C(max)) for both R warfarin (AUC(infinity) [0.93, 1.03], AUC(t) [0.95, 1.03], C(max) [0.90, 1.04]) and S-warfarin (AUC(infinity) [0.93, 1.03], AUC(t) [0.94, 1.03], C(max) [0.90, 1.06]). CONCLUSIONS: The encapsulated form of warfarin (Coumadin) used in comparator studies with the oral DTI ximelagatran is bioequivalent to the nonencapsulated, commercially available form of warfarin (Coumadin). Thus, the results of ximelagatran clinical trials with encapsulated warfarin can be generalized to the commercially available form. PMID- 15751005 TI - Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye. AB - To identify the proteomic alterations associated with carcinogenesis of hepatocellular carcinoma (HCC), we compared the protein expression profiles of nine HCC cell lines with those of primary cultured hepatocytes established from five individuals. A differential proteomic study was performed by two-dimensional difference gel electrophoresis, in which protein samples are labeled with different fluorescent dyes and separated according to their isoelectric point and molecular weight. To label the protein samples, we used a newly developed and highly sensitive fluorescent dye, which reacts with all reduced cysteine residues of proteins. Principal component analysis based on the intensity of 1238 protein spots indicated that the HCC cells and the normal hepatocytes had distinct proteomic profiles. The Wilcoxon test was used to determine the protein spots whose intensity was differentially regulated in the HCC cells compared with the normal hepatocytes, and mass spectrometric analysis was used to identify the proteins corresponding to the spots. The proteins identified are involved in cell cycle regulation, binding to a tumor-suppressor gene product, fatty acid binding, and regulation of translation. Western blotting with specific antibodies revealed the overexpression of PCNA, EB1 and E-FABP in HCC tissues compared with noncancerous tissues. Aberrant regulation of EB1 and E-FABP has not previously been implicated in the development of HCC. PMID- 15751006 TI - Effects of DL-methionine on hatching and activity of Meloidogyne incognita eggs and juveniles. AB - Several concentrations of DL-methionine were tested in the laboratory for their effects on Meloidogyne incognita (Kofoid & White) Chitwood egg hatching and juvenile activity in aqueous suspensions and infested soil. After 7 days in methionine solutions, the proportions of hatched eggs were reduced by 23.3% at 0.25 mg litre(-1) methionine and by 76.4% at 25 g litre(-1) when compared with controls in tap water. An effect of methionine solutions on juvenile activity was also apparent after 24 h in the 25 g litre(-1) treatment, where the percentage of active M incognita juveniles was reduced by 16.3%. Further reductions in nematode mobility were observed as the time of exposure increased, and at lower methionine concentrations over longer exposure times. When methionine solutions were applied to soil infested with M incognita, reductions in egg hatching and juvenile activity were observed at 0.1 and 1 mg methionine g(-1) soil in both sand and clay-loam soils. The percentage of hatched eggs one week after methionine application at rates of 1 mg g(-1) was lower in sand (5.6%) and clay-loam soil (20.8%) than in controls where egg hatch was 52.0% and 48.0% in sand and soil, respectively. Non-active juveniles were found at 1 mg methionine g(-1) soil one week after its application to soil. PMID- 15751007 TI - Effect of chlorpyrifos and monocrotophos on locomotor behaviour and acetylcholinesterase activity of subterranean termites, Odontotermes obesus. AB - The acute toxicity of chlorpyrifos and monocrotophos to subterranean termites, Odontotermes obesus (Rambur), has been studied by a paper contact method. The LC50 values for chlorpyrifos and monocrotophos were 0.046 and 0.148 microg cm( 2), respectively. Chlorpyrifos was 3.22-fold more toxic than monocrotophos. The effect of the pesticides on locomotor behaviour (velocity) and head acetylcholinesterase (AChE: EC 3.1.1.7) activity was estimated in LC50-exposed termites at intervals of 4, 8, 12, 16, 20 and 24 h. Chlorpyrifos- and monocrotophos-treated termites showed, respectively, 97 and 88% reduction in locomotor behaviour (velocity) after 24 h. At all time intervals the chlorpyrifos treated termites exhibited more AChE inhibition and showed greater distorted behaviour than those exposed to monocrotophos. In vitro studies indicated that the I50 value (50% inhibition) for chlorpyrifos against AChE was 8.75 times that of monocrotophos. PMID- 15751008 TI - Synthesis, crystal structure and herbicidal activity of mimics of intermediates of the KARI reaction. AB - Two mimics of the intermediate in the reaction catalyzed by ketol-acid reductoisomerase (KARI) were synthesized. Their structures were established on the basis of elemental analyses, IR, 1H NMR and GC/mass detector. The crystal structure of compound 2 was found to be a substituted dioxane, formed by the condensation of two molecules. The two compounds showed some herbicidal activity on the basis of tests using rape root and barnyard grass growth inhibition. However, the herbicidal effect was weaker in greenhouse tests. PMID- 15751009 TI - Acidic derivatives of the fungicide fenpiclonil: effect of adding a methyl group to the N-substituted chain on systemicity and fungicidal activity. AB - A new acidic derivative of the fungicide fenpiclonil was synthesized containing a methyl group on the alpha-position of the carboxyl function of N-carboxymethyl-3 cyano-4-(2,3-dichlorophenyl)pyrrole. The phloem mobility of the resulting N-(1 carboxyethyl)-3-cyano-4-(2,3-dichlorophenyl)pyrrole was comparable with that of the former compound, but was higher at external pH 5.0. Unlike the derivatives previously synthesized, it was comparable with fenpiclonil in its fungicidal activity against the pathogenic fungus Eutypa lata. PMID- 15751010 TI - Sorption in reconstituted waxes of homologous series of alcohol ethoxylates and n alkyl esters and their effects on the mobility of 2,4-dichlorophenoxybutyric acid. AB - Equilibrium sorption of n-alkyl esters (dimethyl suberate, diethyl suberate, diethyl sebacate, dibutyl suberate and dibutyl sebacate) and monodisperse alcohol ethoxylates (diethylene glycol, tetraethylene glycol, pentaethylene glycol, hexaethylene glycol and octaethylene glycol monododecyl ether) between the reconstituted cuticular waxes of Stephanotis floribunda Brongn (Madagascar jasmine) or Hordeum vulgare L (barley) leaves and an external aqueous receptor solution was determined. Logarithms of the wax/receptor partition coefficient (K(wax/rec)) of the n-alkyl esters increased linearly with the number of C-atoms. With alcohol ethoxylates, log K(wax/rec) decreased linearly with the number of ethylene oxide units. For both groups of compounds, K(wax/rec) increased with increasing lipophilicity. The values of K(wax/rec) in Stephanotis wax were between 5 and 16 times higher than in barley wax. It is argued that this difference was due to different chemical composition and crystallinity of the waxes. Mobility of [14C]2,4-dichlorophenoxybutyric acid (2,4-DB) in reconstituted Stephanotis and barley wax was increased by a factor of 2-8 by both n-alkyl esters and alcohol ethoxylates. Effects on the mobility of 2,4-DB were linearly related to the internal concentrations of n-alkyl esters and alcohol ethoxylates in reconstituted Stephanotis or barley wax. At the same internal concentrations the effect of n-alkyl esters on the mobility of 2,4-DB in wax exceeded that of alcohol ethoxylates by between 1 and 2 orders of magnitude. Results are discussed in relation to formulating systemic pesticides. PMID- 15751011 TI - Time-concentration-mortality modeling of the synergistic interaction of Beauveria bassiana and imidacloprid against Nilaparvata lugens. AB - Interactions between the entomopathogenic fungus, Beauveria bassiana SG8702, and imidacloprid on Nilaparvata lugens (Homoptera: Delphacidae) were studied in laboratory bioassays by spraying suspensions of unformulated conidia (assay 1) and aqueous dilutions of emulsifiable conidia formulation alone (assay 2) or together with imidacloprid at the sub-lethal rates of 0.5, 1.0 and 2.0 microg ml( 1) (assays 3-5). Each assay consisted of five conidia concentrations plus an appropriate control and three replicates, each including 30-40 third-instar nymphs, so as to generate time-concentration-mortality data for modeling analysis. A mineral oil-based emulsion used to formulate B bassiana slightly enhanced fungal activity but had no significant impact on the background mortality of N lugens. On the basis of LC50 estimates and associated variances on days 4-12 after spraying, synergistic interactions of both agents or formulations were determined by estimating relative potencies of assay 2 over assay 1 (1.2 9.0), assay 3 over assay 2 (1.3-1.7), assay 4 over assay 2 (7.5-9.6), assay 5 over assay 2 (22.7-101), assay 4 over assay 3 (3.8-5.8), assay 5 over assay 3 (16.1-61.0), and assay 5 over assay 4 (3.0-10.5). The time-concentration mortality modeling method was not only mathematically but also biologically robust to evaluate the interactions of B bassiana and imidacloprid on N lugens. Compared with their counterparts, enhanced fungal formulations displayed consistently earlier or greater activities against the pest species based on LC50 and LT50 estimates determined from their time-concentration-mortality relationships. The results highlight a potential for pest control by combined formulation or application of B bassiana and imidacloprid. PMID- 15751012 TI - Effect of pretilachlor and fenclorim on carbohydrate and protein formation in relation to their persistence in rice. AB - The persistence of pretilachlor applied to rice singly or in combination with the safener fenclorim was investigated in connection with starch, glucose and protein formation. The addition of fenclorim to pretilachlor did not reduce the accumulation of the latter, but reduced its persistence in rice shoots, while the presence of pretilachlor did not affect the persistence of fenclorim, but significantly increased its accumulation in the shoots. Therefore the safening effect of fenclorim consisted of a more rapid detoxification of pretilachlor. Over the period of pretilachlor and fenclorim detoxification, decreases in starch content, partially counterbalanced by increases of free glucose, and decreases in total protein content were observed in pretilachlor-treated shoots; decreases in both starch and free glucose, as well as in total protein content, were observed in fenclorim -treated shoots compared with the untreated controls. The decreases in starch and total glucose appeared to be a direct consequence of reduced glucokinase and ribulose-1,5-bisphosphate carboxylase activity, and the decrease in total protein an indirect consequence of reduced glutamine synthetase and glutamate synthase activity, in response to pretilachlor and fenclorim treatments. PMID- 15751013 TI - Management of five stored-product insects in wheat with pirimiphos-methyl and pirimiphos-methyl plus synergized pyrethrins. AB - Hard red winter wheat was treated with pirimiphos-methyl at 4, 6 and 8 mg kg(-1), synergized pyrethrins at 0.38, 0.75, 1.13 and 1.5 mg kg(-1), and combinations of the two insecticides, to conduct laboratory bioassays against four beetle pests of stored grain, red flour beetle Tribolium castaneum (Herbst), rusty grain beetle Cryptolestes ferrugineus (Stephens), lesser grain borer Rhyzopertha dominica (F), and rice weevil Sitophilus oryzae (L), and one moth pest, Indianmeal moth Plodia interpunctella (Hubner). Beetle adults and P interpunctella larvae survived well on control wheat, producing a large number of progeny (65-1037 insects per container). Kernel damage in control wheat among the insect species ranged from 9 to 99%. On pirimiphos-methyl-treated wheat, mortality of R dominica adults was > or =72%, but that of the other beetle species and P interpunctella larvae was 100%. Progeny were not produced on pirimiphos-methyl-treated wheat, and the kernel damage was negligible (< or =1%). Synergized pyrethrins were ineffective against the five insect pests. Pirimiphos methyl combined with synergized pyrethrins was not superior to pirimiphos-methyl alone against the five insect pests. Pirimiphos-methyl is not registered in the USA for use on wheat, but our results suggest that it could be a viable grain protectant at rates of 4-8 mg kg(-1). PMID- 15751014 TI - Long-term efficacy and safety of zoledronic acid in the treatment of skeletal metastases in patients with nonsmall cell lung carcinoma and other solid tumors. PMID- 15751016 TI - Heme oxygenase-1 activity after excitotoxic injury: immunohistochemical localization of bilirubin in neurons and astrocytes and deleterious effects of heme oxygenase inhibition on neuronal survival after kainate treatment. AB - An increased expression of the inducible form of heme oxygenase (HO), HO-1, is found in the hippocampus after kainate injection, but thus far it is unclear whether the HO-1 is enzymatically active. The present study was carried out, using monoclonal antibodies to bilirubin and HO-1 and histochemical staining for iron, to compare the products of HO enzymatic activity, bilirubin and iron, with HO-1 expression in the kainate-lesioned hippocampus. There was a close correlation between bilirubin and HO-1 expression, and both bilirubin and HO-1 were observed in damaged neurons at early times, and astrocytes at later times (weeks), after kainate injection. These results indicate that the increased HO-1 in the hippocampus is enzymatically active. Too determine whether HO-1 activity after kainate could have a protective or, perhaps, destructive effect, kainate injected rats were injected intraperitoneally with a blood-brain barrier permeable inhibitor of HO, tin protoporphyrin (SnPP), and the effects of such treatment were compared with effects in rats that received kainate and saline injection. It was found that SnPP treatment did not improve neuronal survival. Instead, increased mortality was observed in rats treated with SnPP. Four SnPP injected rats vs. one saline-injected rats died after kainate treatment. The surviving SnPP-treated rats showed significantly less hippocampal field that containing Nissl or MAP2 staining (an indicator of surviving neurons) compared with the saline-injected rats. These results indicate that HO-1 induction had a net protective effect on neurons in the kainate model of excitotoxic injury. PMID- 15751017 TI - Prediction of survival in patients with esophageal carcinoma using artificial neural networks. AB - BACKGROUND: Accurate estimation of outcome in patients with malignant disease is an important component of the clinical decision-making process. To create a comprehensive prognostic model for esophageal carcinoma, artificial neural networks (ANNs) were applied to the analysis of a range of patient-related and tumor-related variables. METHODS: Clinical and pathologic data were collected from 418 patients with esophageal carcinoma who underwent resection with curative intent. A data base that included 199 variables was constructed. Using ANN-based sensitivity analysis, the optimal combination of variables was determined to allow creation of a survival prediction model. The accuracy (area under the receiver operator characteristic curve [AUR]) of this ANN model subsequently was compared with the accuracy of the conventional statistical technique: linear discriminant analysis (LDA). RESULTS: The optimal ANN models for predicting outcomes at 1 year and 5 years consisted of 65 variables (AUR = 0.883) and 60 variables (AUR = 0.884), respectively. These filtered, optimal data sets were significantly more accurate (P < 0.0001) than the original data set of 199 variables. The majority of ANN models demonstrated improved accuracy compared with corresponding LDA models for 1-year and 5-year survival predictions. Furthermore, ANN models based on the optimal data set were superior predictors of survival compared with a model based solely on TNM staging criteria (P < 0.0001). CONCLUSIONS: ANNs can be used to construct a highly accurate prognostic model for patients with esophageal carcinoma. Sensitivity analysis based on ANNs is a powerful tool for seeking optimal data sets. PMID- 15751019 TI - Study on the morphology and reproducibility of the diagnosis of endometrial lesions utilizing liquid-based cytology. AB - BACKGROUND: The objective of the current study was to determine the diagnostic cytomorphologic criteria for liquid-based cytology and to evaluate the reproducibility and usefulness of the cytologic diagnosis in endometrial lesions. METHODS: A total of 162 direct endometrial samplings taken from postmenopausal women were evaluated by 2 skilled cytopathologists in endometrial cytology. The cytologic diagnosis was made according to the 1994 classification scheme of the World Health Organization. After establishment of the criteria, three additional cytopathologists without any experience in liquid-based endometrial cytology examined the same cases to determine interobserver variability. The intraobserver variability also was evaluated by all the observers. RESULTS: The cytomorphologic criteria were established in the following four diagnostic categories: atrophic endometrium, hyperplasia without atypia, hyperplasia with atypia, and adenocarcinoma. The overall interobserver agreement was nearly perfect with a kappa value of 0.89 during the checking round and ranged from moderate to substantial with kappa values of 0.48-0.80, respectively, in the other diagnostic rounds (P < 0.0001); hyperplasia with atypia was found to be the most difficult category to identify correctly. Furthermore, the intraobserver agreement ranged from substantial to perfect with kappa values of 0.61-1.00 in all diagnostic rounds (P < 0.0001). CONCLUSIONS: Liquid-based cytology allows for standardized and reproducible endometrial preparations, which in turn allows the application of common diagnostic criteria among cytopathologists. Furthermore, liquid-based cytology in combination with endometrial sampling could be a useful tool for the outpatient diagnosis of endometrial lesions, which could reduce unnecessary curettage. PMID- 15751018 TI - Clinicopathologic significance of dysadherin expression in cutaneous malignant melanoma: immunohistochemical analysis of 115 patients. AB - BACKGROUND: The E-cadherin-mediated cell adhesion system is frequently inactivated by multiple mechanisms and is involved in tumor progression in many types of cancer. Recently, the authors reported a novel cell membrane glycoprotein, dysadherin, which has an anti-cell-cell adhesion function and down regulates E-cadherin. METHODS: Expression of both dysadherin and E-cadherin was investigated immunohistochemically in 115 patients with cutaneous malignant melanoma to determine the correlation between the 2 molecules and their associations with both patient survival and the clinicopathologic features of the tumors. RESULTS: Dysadherin and E-cadherin were expressed at the cell membranes of melanoma cells. Fifty-two percent of the tumors showed dysadherin immunopositivity, and 91% of the tumors showed reduced E-cadherin immunopositivity. There was no significant inverse correlation between dysadherin expression and E-cadherin expression. Increased dysadherin expression was significantly correlated with nodular subtype (P = 0.042), Clark level (P < 0.001), tumor thickness (P < 0.001), ulceration (P = 0.008), lymph node metastasis (P < 0.001), high TNM classification (P < 0.001), and poor patient survival (P < 0.001). Multivariate analysis of patient survival revealed that increased dysadherin expression was a significant predictor of poor survival (P < 0.001). CONCLUSIONS: Thus, increased expression of dysadherin was a significant indicator of poor prognosis in patients with cutaneous malignant melanoma. PMID- 15751020 TI - Lack of trastuzumab activity in nonsmall cell lung carcinoma with overexpression of erb-B2: 39810: a phase II trial of Cancer and Leukemia Group B. AB - BACKGROUND: The overexpression of HER-2 occurs in a minority of patients with nonsmall cell lung carcinoma. Trastuzumab, which is a monoclonal antibody to HER 2, is an effective treatment in some women with breast carcinomas that overexpress HER-2, as demonstrated by immunohistochemistry. The objective of this Phase II study was to determine whether trastuzumab would effect responses in patients with nonsmall cell lung carcinoma who had tumors that overexpressed HER 2. METHODS: Patients were required to have Stage IIIB or Stage IV nonsmall cell lung carcinoma and tumors with 2+ or 3+ expression of HER-2, as determined with immunohistochemistry, and they may have received up to 1 prior chemotherapy regimen. Trastuzumab at a dose of 4 mg/kg was given intravenously on Week 1; then, weekly doses of 2 mg/kg were given. Response revaluation was performed every 8 weeks. RESULTS: Among 209 screened patients, 24 patients (11%) had tumors with 2+ or 3+ expression of HER-2. One patient achieved a partial response, and one patient experienced a treatment-related death due to pulmonary toxicity. CONCLUSIONS: Single-agent trastuzumab did not exhibit significant clinical activity against nonsmall cell lung carcinoma when HER-2 expression levels were measured by immunohistochemistry. PMID- 15751021 TI - 'Objective' determination of coupling constants and conformational equilibria by solvent variation: a re-evaluation. AB - An 'objective' method for determining conformational equilibria in substituted ethanes, proposed by Abraham et al., has been evaluated by computational methods. Abraham's method involves measuring vicinal couplings, such as (3)J(H,H) and (3)J(H,F), between methine and methylene protons with methine, methylene protons and fluorine in a range of solvents, on the assumption that the underlying coupling constants of the individual conformers are constant, but the fractions of each conformer in each solvent are different and unknown. Abraham posited that this would produce an 'over-determined' data set with more equations than unknowns would. Abraham's procedure is re-evaluated, and it is demonstrated that the type of system being considered here, where there are more equations than unknowns, is not necessarily over-determined. A computer equation solver and Monte Carlo-type procedures were employed to demonstrate that multiple numerical solutions exist for a representative 'over-determined' data set provided by Abraham. A statistical method was also developed to determine precisely which parameter sets constitute plausible solutions. PMID- 15751022 TI - Synthesis and NMR spectral study of some t(3)-aryl-r(2),c(4)-bisethoxycarbonyl t(5)-hydroxy-c(5)-methylcyclohexanones. AB - Six t(3)-aryl-r(2),c(4)-bisethoxycarbonyl-t(5)-hydroxy-c(5)-methylcyclohexanones (6-11) were synthesized by condensing ArCHO (Ar = Ph, p-O(2)NC(6)H(4), p CH(3)OC(6)H(4), p-ClC(6)H(4), m-O(2)NC(6)H(4) and m-C(6)H(5)O(6)H(4)) with ethyl acetoacetate in the presence of methylamine and their (1)H and (13)C NMR spectra were recorded. (1)H-(1)H COSY and NOESY spectra were recorded for 6 and 7 and also HSQC and HMBC spectra for 6 and 8. Elemental analysis was carried out for all compounds. The mass spectrum was recorded for 8. All analytical data are consistent with the proposed molecular formulae. Analysis of NMR spectral data suggests that these compounds largely adopt chair conformations with the hydroxyl group occupying an axial orientation and all the other substituents occupying equatorial orientations. Long-range coupling (2-3 Hz) between the OH proton and the axial methylene proton at C-6 is observed in 6, 7, 8 and 11. PMID- 15751023 TI - 'Non-covalent synthesis' of a chiral host of calix[6]arene and enantiomeric discrimination. AB - 'Non-covalent synthesis' of novel chiral hosts (calix[6]arene-chiral amine complexes) and its application to enantiomeric discrimination was investigated by (1)H NMR spectroscopy. The topology of a ternary complex was proposed for the calix[6]arene-amine-sulfoxide to rationalize the chiral recognition. PMID- 15751024 TI - Incidence, cost, and mortality of neutropenia hospitalization associated with chemotherapy. AB - BACKGROUND: Neutropenia is a common side effect of chemotherapy, often requiring hospitalization for treatment of severe cases. Neutropenia hospitalization (NH) rates have been reported in individual studies, but national estimates are needed. METHODS: Chemotherapy-induced NHs were identified in the 1999 hospital discharge data bases from 7 states. Cancer and chemotherapy prevalence data from the National Cancer Institute's Surveillance, Epidemiology, and End Results Program and the National Cancer Data Base were used to calculate national NH rates for 13 cancer types. NH cost was estimated by multiplying charges by institution-specific, cost-to-charge ratios from the 1999 Centers for Medicare and Medicaid Services Hospital Cost Report. NH incidence was projected to national levels using population data from the United States Census and the Centers for Disease Control and Prevention. RESULTS: There were 20,780 discharges with documentation of cancer, chemotherapy, and neutropenia identified. Projecting to national levels, NH incidence was estimated at 60,294 cases (7.83 cases per 1000 cancer patients). The mean NH cost was 13,372 dollars. The mortality rate among patients with NH was estimated at 6.8% or 1 death for every 14 hospitalized patients. Among 13 selected cancer types, the NH rate was 34.20 cases per 1000 patients receiving chemotherapy (1 in 29 patients). NH was particularly common in patients with hematologic tumors, with an incidence of 43.3 cases per 1000 patients with such tumors (1 in 23 patients). The average NH cost for hematologic malignancies was 20,400 dollars, more than double the cost of NH for solid tumors. CONCLUSIONS: According to the current study, NH affects > 60,000 patients with cancer each year in the United States, with an average cost of 13,372 dollars per hospitalization and an associated inpatient mortality rate of 6.8%. PMID- 15751025 TI - Shaw potassium channel genes in Drosophila. AB - Drosophila Shaw encodes a voltage-insensitive, slowly activating, noninactivating K(+) current. The functional and developmental roles of this channel are unknown. In this study, we use a dominant transgenic strategy to investigate Shaw function and describe a second member of the Shaw family, Shawl. In situ hybridization showed that the two Shaw family genes, Shaw and Shawl, have largely nonoverlapping expression patterns in embryos. Shaw is expressed mainly in excitable cells of the CNS and PNS of late embryos. Shawl is expressed in many nonexcitable cell types: ubiquitously in embryos until the germband extends, then transiently in the developing CNS and PNS, becoming restricted to progressively smaller subsets of the CNS. Ectopic full-length and truncated Shaw localize differently within neurons, and produce uneclosed small pupae and adults with unfurled wings and softened cuticle. This phenotype was mapped to the crustacean cardioactive peptide (CCAP)-neuropeptide circuit. Widespread expression of Shaw in the nervous system results in a reduction in body mass, ether-induced shaking, and lethality. Expression of full-length Shaw had more extreme phenotypic consequences and caused earlier lethality than expression of truncated Shaw in a given GAL4 pattern. Whole cell recordings from ventral ganglion motor neurons expressing the truncated Shaw protein suggest that a major role of Shaw channels in these cells is to contribute to the resting potential. PMID- 15751026 TI - A group of neurokinin-1 receptor-immunoreactive neurons expressing phospho extracellular signal-regulated protein kinases in the pre-Botzinger complex of rats. AB - Extracellular-signal-regulated kinases (ERKs) were described originally as critical regulators of cell growth and differentiation. ERKs have received increasing attention as important regulators of neuronal function, especially in synaptic plasticity and memory. We found a group of phospho-ERK1/2 (pERK1/2) immunoreactive (ir) neurons in the ventrolateral medulla oblongata of adult rats. In the pre-Botzinger complex (pre-BotC) region, identified by neurokinin-1 receptor (NK1R) immunoreactivity, the distribution of pERK1/2-ir neurons seemed to overlap with that of NK1R-ir neurons. A detailed study combining immunofluorescence and immunogold-silver staining was designed to determine whether ERK-ir neurons were NK1R-ir and related to respiratory function in the pre-BotC. The pERK1/2 immunoreactivity was present mainly in somas and processes of small- to medium-sized neurons. Of pERK1/2-ir neurons in the pre-BotC, 19.7% were NK1R-ir. Some of the small, fusiform, NK1R-ir neurons, the putative rhythmogenic neurons, were also pERK1/2-ir. Under the electron microscope, immunogold particles indicative of pERK1/2 immunoreactivity were found in somas and dendrites in pre-BotC neurons. They were associated mainly with rough endoplasmic reticula and ribosomes. Immunogold particles could also be found in nuclei, but were absent from nucleoli. The existence of pERK1/2 in a subtype of NK1R-ir pre-BotC neurons implies that the pERK cascade may play a role in normal respiratory activity in adult rats. The pERK cascade may also participate in sympathoexcitatory activity, given that 42.2% of pERK1/2-ir neurons expressed tyrosine hydroxylase, a marker of C1 adrenergic neurons in the ventrolateral medulla oblongata. PMID- 15751027 TI - Migration and fate of newly born cells after focal cortical ischemia in adult rats. AB - Neural cell migration and differentiation may participate in neural repair after adult brain injury; however, the survival and differentiation of newly born cells after different brain lesions are poorly understood. We have examined the migration and fate of bromodeoxyuridine (BrdU)-labeled cells after a highly reproducible focal ischemic lesion restricted to the frontoparietal cortex in adult rats. Thermocoagulation of pial blood vessels induces a circumscribed degeneration of all cortical layers while sparing the corpus callosum and striatum and increases cell proliferation in the subventricular zone (SVZ) and rostral migratory stream (RMS) within 7 days. We now show that, although the rostral migration of the newly born SVZ cells and their differentiation into neurons in the olfactory bulb were not affected by the lesion, numerous cells expressing the neuroblast marker doublecortin migrated laterally in the striatum and corpus callosum 5 days postinjury. In addition to the SVZ, BrdU-labeled cells were seen in the striatum, in the corpus callosum, and around the lesion. One month later, BrdU-labeled cells in the corpus callosum expressed transferrin and the pi isoform of glutathione-S-transferase (GST-pi), markers of oligodendrocytes. Other BrdU+ cells expressed a marker of astrocytes, but none expressed neuronal markers, suggesting that new neurons do not form or survive under these conditions. Numerous BrdU-labeled cells were still observed in the SVZ and RMS. The data show that focal cortical ischemia does not lead to the long term survival of new neurons in the striatum or cortex but induces long-term alterations in the SVZ and the production of new oligodendrocytes that may contribute to neural repair. PMID- 15751028 TI - Lack of large intestinal carcinogenicity of 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine at low doses in rats initiated with azoxymethane. AB - 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), an abundant food-derived heterocyclic amine (HCA), has attracted particular attention as a human colon carcinogen. Humans are in fact exposed to continuous low doses of HCAs during lifetime. Therefore, we focused on rat large intestinal carcinogenicity of PhIP at levels that mimic practical human exposure. A total of 192 6-week-old male F344 rats were subcutaneously injected twice with 15 mg/kg body weight azoxymethane (AOM), then continuously fed various doses (0, 0.001, 0.01, 0.1, 1, 10, 50 and 200 ppm) of PhIP in the diet. At week 16, aberrant crypt foci (ACF) were quantitatively analyzed. At week 36, tumor occurrence was pathologically analyzed. Then immunohistochemical examinations were performed. PhIP was found to enhance strongly AOM-initiated rat large intestinal tumorigenesis at high doses (50 and 200 ppm), while lower doses (0.001-10 ppm) had no apparent effects. High doses also caused variation in tumor histologic types and their distribution throughout the large intestinal segments. Frequencies of ACF/cm(2) did not meaningfully vary between the groups. Cellular proliferation activity in normal appearing colonic mucosa was significantly increased at high doses. These novel findings may provide evidence of a low-dose potential for PhIP, with a no observed effect level speculated to be 10 ppm in the present initiation-promotion experimental model. PMID- 15751029 TI - Using finite-element analysis to investigate suture morphology: a case study using large carnivorous dinosaurs. AB - Finite-element analysis (FEA) can be used to investigate the mechanical significance of sutures and regions of intracranial flexibility in skulls. By modeling the stress response to feeding forces in a finite-element skull model (with appropriate boundary conditions), one can compare the axis of distortion and orientation of stress and strain in the model to the degree of movement at actual sutural contacts in the real skull. Hypotheses detailing the effect of introducing patency or flexibility on mechanical performance can be constructed and subsequently tested. In this study, the correlation between stress environment, cranial strength, and sutural morphology and mobility is investigated in the cranium of the large theropod dinosaur Allosaurus fragilis. Theropods are an especially interesting model system as their skulls were massive (over 100 cm in some cases), may have generated extremely large bite forces, yet patent sutures persisted between many of the facial bones. In this analysis, it was discovered that Allosaurus cranial sutures appear generally capable of accommodating stress and strain patterns generated during biting. This study highlights the potential of FEA in devising and testing hypotheses of form and function and argues that useful information can be obtained from finite-element models of extinct animals, providing that adequate assumptions are made and appropriate questions asked. PMID- 15751030 TI - Diethylstilbestrol exposure in utero: a paradigm for mechanisms leading to adult disease. AB - The synthetic estrogen diethylstilbestrol (DES) was administered to pregnant women between the 1940s and the mid-1970s and is believed to be responsible for numerous uterine/cervical/vaginal malformations and cancers that appeared after birth and in young adult life. This medical tragedy has served as one of the prototypical examples of a phenomenon known as "endocrine disruption," in which either environmental agents or other compounds disrupt normal hormonal signaling in the body. Whereas DES signals through estrogen receptors, the subsequent molecular targets were largely unknown. We had identified Wnt7a as a target in this pathway and have used genetic analyses of mutant mice to demonstrate that disruption of Wnt7a is the key event leading to the DES phenotypes and cancers. We find that Wnt7a expression is only transiently deregulated in response to DES exposure, leading to the conclusion that critical events during early reproductive tract development results in a permanent change or "reprogramming" in subsequent development. PMID- 15751031 TI - Knowledge and use of papanicolaou test among HIV-positive women. PMID- 15751032 TI - Effect of the tumor promoter phenobarbital on the pattern of global gene expression in liver of connexin32-wild-type and connexin32-deficient mice. AB - The antiepileptic drug phenobarbital (PB) is used frequently as a model tumor promoter in rodent liver. It is believed to increase the probability of cancer by accelerating the clonal expansion of cells transformed during tumor initiation. The molecular mechanism underlying this process is only partly understood but seems to require the function of connexin32 (Cx32), one of the 2 gap junction proteins expressed in hepatocytes. PB mediates transcriptional activation of various genes in liver but which of these are relevant for tumor promotion is unknown. We have used oligonucleotide microarrays to identify genes differentially modulated in expression by PB in liver of Cx32-wild-type and Cx32 null mice. Mice of both strains were kept on PB containing (0.05%) or control diet for 2 weeks. Total liver RNA was isolated from 3 mice per experimental group and reverse transcribed; cDNAs were hybridized to oligonucleotide microarrays and a gene-by-gene linear model was used for statistical analysis of data. Five genes were identified as induced or repressed in untreated Cx32-null as compared to untreated Cx32-wild-type mice. PB affected the expression of 53 genes, of which 13 code for members of Phase-I/II of drug metabolism, and 12 genes were differentially affected in expression by PB in Cx32-null as compared to Cx32-wild type mice. Among the differentially affected genes that could be verified by quantitative RT-PCR or Western analysis were the insulin like growth factor binding protein-1, retinol dehydrogenase-6 and the Y-chromosomally located gene Dby, among which may be a candidate of relevance for PB-mediated tumor promotion. PMID- 15751033 TI - NY-ESO-1/LAGE-1 coexpression with MAGE-A cancer/testis antigens: a tissue microarray study. AB - The characterization of the expression pattern of different families of cancer/testis (C/T) antigens in different tumors, at the protein level, might be of relevance in the development of multiantigen vaccine preparations for active specific immunotherapy. We have used tissue microarray (TMA) technology to explore in large numbers of tumor specimens the expression of NY-ESO-1/LAGE-1 C/T antigens and its correlation with MAGE-A expression by using D8.38 and 57B monoclonal antibodies (MAb). The epitopes recognized by these reagents in C/T antigens were identified by molecular mapping by using a bacterial expression system. Out of 2,052 samples, 119 (5.8%) scored positive upon staining with D8.38 NY-ESO-1/LAGE-1-specific MAb. Expression in >10% of cases was detectable in melanoma and basalioma (31.6 and 18.2%, respectively), large cell carcinomas and adenocarcinomas of the lung (17.8 and 10.5%, respectively), stomach adenocarcinomas of the intestinal type (13.2%), pT2-4 bladder TCC (18.2%), nonseminomatous carcinomas of the testis (10.4%) and liposarcomas (15.4%). Simultaneous expression of NY-ESO-1/LAGE-1 and MAGE-A C/T antigens was then addressed in a TMA where 101/845 and 73/845 samples (12 and 8.6%, respectively) showed evidence of MAGE-A or NY-ESO-1/LAGE-1 specific staining, respectively. In 35/845 specimens (4.1%) concomitant expression of MAGE-A and NY-ESO-1/LAGE-1 was observed (p = 0.0002). Discrepancies in the expression of NY-ESO-1/LAGE-1 and MAGE-A were conspicuously detectable in squamous cell carcinomas of the skin (MAGE-A positive but NY-ESO-1/LAGE-1 negative) and in liposarcomas (NY-ESO-1/LAGE 1 positive, but MAGE-A negative). Taken together, these data suggest novel areas of application of C/T antigens targeted active specific immunotherapy possibly based on multiantigen vaccine preparations. PMID- 15751034 TI - A de novo, apparently balanced reciprocal translocation in a child with developmental delay whose mother was being treated with low-dose methotrexate at the time of conception. AB - BACKGROUND: Methotrexate is a proven teratogen and its use periconceptually is strongly advised against. Concerns about possible chromosomal effects in the offspring of women treated with this agent have been raised, but they have not been reported. In vivo and in vitro studies have supported this possibility, however. CASE: A 32-year-old primigravida was treated with low-dose methotrexate for rheumatoid arthritis from prior to conception until six weeks postconception. Her child was born without congenital malformations but subsequently developed seizures and was diagnosed with developmental delay. He was found to have a de novo, apparently balanced, reciprocal translocation between chromosomes 5 and 20 (46,XY,t(5:20)(q15;p12)). Other investigations failed to reveal another cause for his developmental delay. CONCLUSIONS: The possible association between maternal methotrexate exposure and a de novo chromosomal anomaly in an offspring supports the recommendation that women should be advised to cease taking this drug for at least six months prior to conception. PMID- 15751036 TI - Genesis of derivative chromosome 9 deletions in chronic myeloid leukemia. PMID- 15751035 TI - Epigenetic silencing of the imprinted gene ZAC by DNA methylation is an early event in the progression of human ovarian cancer. AB - ZAC is a paternally expressed, imprinted gene located on chromosome 6q24, within a region known to harbor a tumor suppressor gene for several types of neoplasia, including human ovarian cancer (HOC). We have failed to identify genetic mutations in the ZAC gene in tumor material. Many imprinted genes contain differentially allele-specific-methylated regions (DMR) and harbor promoter activity that is regulated by the DNA methylation. Aberrant DNA methylation is a common feature of neoplasia and changes in DNA methylation at the ZAC locus have been reported in some cases of HOC. We investigated the DNA methylation and ZAC mRNA expression levels in a larger sample of primary HOC material, obtained by laser capture microdissection. ZAC mRNA expression was reduced in the majority of samples and this correlated with hypermethylation of the ZAC-DMR. Treatment of hypermethylated cells lines with a demethylating agent restored ZAC expression. Our studies indicate that transcriptional silencing of ZAC is likely to be caused by DNA methylation in HOC. Forced expression of ZAC resulted in a reduction in proliferation and marked induction of apoptotic cell death. The ZAC-mediated apoptosis signal is p53-independent and eliminated by inhibitors of caspase 3, 8 and 9. Reduced expression of ZAC would therefore favor tumor progression. As there were no significant differences in either DNA methylation or expression of ZAC mRNA between localized and advanced tumors, our data indicates that loss of ZAC is a relatively early event in HOC. (Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.) PMID- 15751038 TI - The fetal basis of adult disease: Role of environmental exposures--introduction. PMID- 15751039 TI - Developmental pesticide exposures and the Parkinson's disease phenotype. AB - Whereas Parkinson's disease is a neurodegenerative disorder that typically onsets after 60 years of age, the possibility that it could result from insults sustained during development has been proposed. Experimental evidence based on the combined paraquat + maneb model of the Parkinson's disease (PD) phenotype summarized here provides support for such an assertion. Postnatal exposures of mice to these pesticides led not only to a permanent and selective loss of dopaminergic neurons in the substantia nigra pars compacta but also enhanced the impact of these pesticides administered during adulthood relative to developmental only or adult only treatment. Exposure to maneb alone during gestation resulted in a dramatic response to paraquat in adulthood, including notable reductions in levels of dopamine and metabolites and a loss of nigral dopamine (DA) neurons, despite the fact that paraquat does not share structural similarity to or mechanisms of action with maneb. Collectively, these studies provide developmental environmental models of the PD phenotype. In addition, they demonstrate the fact that silent neurotoxicity produced by developmental insults can be unmasked by challenges later during life as well as the potential for cumulative neurotoxicity over the life span. PMID- 15751040 TI - MLL is fused to EB1 (MAPRE1), which encodes a microtubule-associated protein, in a patient with acute lymphoblastic leukemia. AB - We have shown that the EB1 (MAPRE1) gene, at 20q11.2, is fused to MLL in an adult patient with pro-B acute lymphoblastic leukemia. Southern blot analysis indicated that a rearrangement of the MLL gene was involved in the chromosomal abnormality. cDNA panhandle polymerase chain reaction (PCR) identified the fusion transcript, in which MLL exon 6 was fused in-frame with EB1 exon 5. The presence of the MLL EB1 and the reciprocal EB1-MLL fusion transcripts was verified by reverse transcription PCR. EB1 is the first gene on chromosome 20 found to fuse with MLL. The genomic break junctions of MLL-EB1 and EB1-MLL were amplified by long distance PCR. Sequencing of the break junctions revealed that multiple DNA breaks had occurred and that the DNA fragments flanked by these breaks had been duplicated, deleted, or inverted. Nontemplate DNA segments of 2 bp also were detected at the breakpoints on derivative chromosomes 11 and 20. These features indicate that this translocation likely resulted from the DNA damage-repair pathway. EB1 is a microtubule-associated protein that interacts with the colorectal adenomatous polyposis coli tumor-suppressor protein and plays important roles in regulating microtubule dynamics, cell polarity, and chromosome stability. Immunofluorescence staining demonstrated that the MLL-EB1 fusion proteins were localized in the nuclei. PMID- 15751041 TI - Genetic and epigenetic alterations of the candidate tumor-suppressor gene MYO18B, on chromosome arm 22q, in colorectal cancer. AB - Allelic imbalance (AI) on chromosome arm 22q has been detected in 20%-40% of colorectal cancers, suggesting that this chromosome arm has a tumor-suppressor gene involved in colorectal carcinogenesis. Recently, we isolated a candidate tumor-suppressor gene, MYO18B, at 22q12.1, that is deleted, mutated, and hypermethylated in more than 50% of lung cancers. In the present study, we analyzed genetic and epigenetic alterations of the MYO18B gene in colorectal cancers. AI at the MYO18B locus was detected in 16 of 43 (40%) informative cases. Mutations of the MYO18B gene were detected in 2 of 11 (18%) cell lines and 1 of 47 (2%) surgical specimens. Nine of 11 (82%) cell lines showed reduced MYO18B expression, which was restored in all 9 by treatment with 5-aza-2'-deoxycytidine and/or trichostatin A (TSA). Although hypermethylation of the promoter CpG island for MYO18B was not detected, a significant correlation was observed between the level of MYO18B expression and the level of acetylation of histones H3 and H4 in 6 cell lines with and without TSA treatment. Thus, it was suggested that MYO18B is inactivated in a considerable fraction of colorectal cancers by several mechanisms, especially silencing by histone deacetylation and/or AI. Furthermore, restoration of MYO18B expression in colorectal cancer cell lines HT29 and DLD-1 suppressed anchorage-independent growth, whereas it did not affect the growth rate in vitro. These results suggest that genetic and epigenetic inactivation of the MYO18B gene play an important role in colorectal carcinogenesis. PMID- 15751042 TI - Ductal breast carcinoma develops through different patterns of chromosomal evolution. AB - In a previous study that used comparative genomic hybridization (CGH) to analyze 43 ductal breast carcinomas selected for hyperdiploidy, we proposed the existence of two distinct pathways of chromosomal evolution. In the present study, in which we reassessed our cytogenetic findings on 158 ductal breast carcinomas selected for having a modal number of chromosomes of fewer than 60, we confirmed the existence of two subtypes of tumors. Along with the great majority of tumors (142 of 158) that evolved through structural rearrangements with no or very few whole chromosome gains, we found that a minor subset (16 of 158) evolved through progressive gains of whole chromosomes with no or only a few associated rearrangements. In this article, we describe the karyotypes of these 16 tumors together with data from CGH, which was performed for 10 of them. Chromosomes 5, 7, 8, and 20 were the most frequently gained. Our findings support the evidence of a new pathway of chromosomal evolution in a small subset of ductal breast carcinomas characterized by numerical chromosome aberrations. PMID- 15751043 TI - The importance of appropriate controls, animal feed, and animal models in interpreting results from low-dose studies of bisphenol A. AB - Interpreting results of studies that report only negative effects is problematic. A number of published studies to determine whether chemicals with estrogenic activity can cause effects at low doses have not taken into account the possibility that the commercial animal feed being used can mask effects of even potent estrogenic drugs such as diethylstilbestrol (DES). In addition, the sensitivity of the strain of animal being used for the specific category of chemical being tested has not always been described. For environmental chemicals, such as the estrogenic polycarbonate plastic monomer bisphenol A, DES is an appropriate positive control for estrogenic effects, and using an appropriate low dose of DES can eliminate the possibility of false-negative conclusions of safety when the above or other variables contribute to the negative outcome. Only when simultaneous positive effects of low doses of a positive control chemical such as DES and negative effects of environmentally relevant low doses of the test chemical are demonstrated within the same experiment are conclusions of no effect of the test chemical warranted, and this has not been reported for bisphenol A in any study. Instead, more than 90 refereed journal publications have reported effects due to exposure to low doses of bisphenol A in a wide variety of animals (for references see: http://rcp.missouri.edu/endocrinedisruptors/vomsaal/vomsaal.html). However, due to lack of attention to the importance of appropriate positive controls, a small number of studies reporting negative effects of bisphenol A have created a false sense of controversy regarding low-dose effects of bisphenol A. PMID- 15751044 TI - Extraosseous IL-6 transgenic mouse plasmacytoma sometimes lacks Myc-activating chromosomal translocation. AB - The cellular oncogene MYC and plasma cell growth, differentiation, and survival factor IL-6 play critical roles in the natural history of human plasma cell neoplasms such as multiple myeloma (MM). Myc and IL-6 also are at the center of neoplastic plasma cell transformation in BALB/c mice that carry a human IL-6 transgene and, therefore, predictably develop plasmacytomas (PCTs). We showed previously that, much like advanced MM or human myeloma cell lines (HMCLs), in which MYC is frequently deregulated in cis because of complex cytogenetic aberrations juxtaposing MYC to immunoglobulin enhancers, IL-6 transgenic PCTs commonly deregulate Myc in cis by chromosomal translocation, predominantly T(12;15)(Igh-Myc). In this article, we show that, analogous to primary MM in which MYC is mostly deregulated in trans by signaling pathways converging at the MYC promoter, IL-6 transgenic PCTs sometimes develop in the absence of Myc translocations, thus activating Myc in trans. We present cytogenetic and molecular evidence on two IL-6 transgenic PCTs that contained overexpressed Myc protein but lacked T(12;15)(Igh-Myc) and two related Myc--deregulating translocations that juxtapose Myc to immunoglobulin light-chain instead of heavy chain enhancers: T(6;15)(Igkappa-Pvt1) and T(15;16)(Pvt1-Iglambda). We conclude that Myc translocations are not strictly required for IL-6-driven PCT development in mice. IL-6 transgenic PCTs may provide a valuable model system for elucidating both trans and cis mechanisms of Myc deregulation of great relevance for MYC deregulation in human MM. PMID- 15751045 TI - Triage using HPV-testing in persistent borderline and mildly dyskaryotic smears: proposal for new guidelines. AB - In the Netherlands 2% of cervical smears in the cervical cancer screening program are read as borderline or mildly dyskaryotic cytology (BMD smear). Only in about 10% of these women a high-grade CIN lesion (CIN II-III) is present; therefore referral is for the majority unnecessary. In our study triage with high-risk HPV (hrHPV) testing was used to identify women at risk for development of high-grade CIN lesions after a repeat BMD smear. A "wait-and-see" period was incorporated allowing clearance of HPV and regression of the lesion. Women with a low-grade lesion, irrespective of their HPV status, were monitored at 12 months; women with a high-grade lesion were monitored at 6 and 12 months. Fifty-one of the 105 women (49%) were hrHPV negative at baseline; none of them showed progression of the lesion within the first year of follow-up (NPV 100%). High-grade CIN was present in 1 patient who was HPV negative at baseline (2%); she demonstrated regression after 12 months. Nineteen of the hrHPV positive women (35%) demonstrated a high grade CIN lesion at baseline and 3 cleared hrHPV after 6 months, with a subsequent regression of CIN. Ten women remained hrHPV positive with persistence of high-grade CIN and were eventually treated. At baseline, 35 hrHPV positive women demonstrated a low-grade lesion, 19 remained hrHPV positive after 12 months and 5 developed high-grade CIN. Sixteen out of the 35 cleared the hrHPV infection without progression of the lesion. In conclusion, triage, using hrHPV testing for women with persistent BMD cytology, can select women who are not at risk for development of high-grade CIN. We recommend return to the screening program without referral for colposcopic examination if hrHPV is absent. For hrHPV positive women, a repeat hrHPV test after another 6 months is suggested. Referral is only required if persistence of hrHPV is established. PMID- 15751046 TI - AML M3 and AML M3 variant each have a distinct gene expression signature but also share patterns different from other genetically defined AML subtypes. AB - Acute promyelocytic leukemia (APL) with t(15;17) appears in two phenotypes: AML M3, with abnormal promyelocytes showing heavy granulation and bundles of Auer rods, and AML M3 variant (M3v), with non- or hypogranular cytoplasm and a bilobed nucleus. We investigated the global gene expression profiles of 35 APL patients (19 AML M3, 16 AML M3v) by using high-density DNA-oligonucleotide microarrays. First, an unsupervised approach clearly separated APL samples from other AMLs characterized genetically as t(8;21) (n = 35), inv(16) (n = 35), or t(11q23)/MLL (n = 35) or as having a normal karyotype (n = 50). Second, we found genes with functional relevance for blood coagulation that were differentially expressed between APL and other AMLs. Furthermore, a supervised pairwise comparison between M3 and M3v revealed differential expression of genes that encode for biological functions and pathways such as granulation and maturation of hematologic cells, explaining morphologic and clinical differences. Discrimination between M3 and M3v based on gene signatures showed a median classification accuracy of 90% by use of 10-fold CV and support vector machines. Additional molecular mutations such as FLT3-LM, which were significantly more frequent in M3v than in M3 (P < 0.0001), may partly contribute to the different phenotypes. However, linear regression analysis demonstrated that genes differentially expressed between M3 and M3v did not correlate with FLT3-LM. PMID- 15751047 TI - Mucin-depleted foci have beta-catenin gene mutations, altered expression of its protein, and are dose- and time-dependent in the colon of 1,2-dimethylhydrazine treated rats. AB - Mucin-depleted foci (MDF) are purported preneoplastic lesions that can be easily visualized in the unsectioned colon of carcinogen-treated rats stained with high iron diamine alcian blue (HID-AB). In F344 rats treated twice with 150 mg/kg of 1,2-dimethylhydrazine (DMH) and sacrificed after 5, 9, 13 and 28 weeks, MDF increased over time from 5 to 13 weeks, whereas they decreased at 28 weeks, when tumors appear. MDF multiplicity (crypts/MDF) linearly increased with time. Increasing doses of DMH (100, 150 and 200 mg/kg x 2 times) caused a dose-related increase in MDF. Mutations in Ctnnb1 gene codifying for beta-catenin were identified with PCR amplification and direct sequencing in 6/15 tumors (40%), 7/28 MDF (25%) and 2/27 (7%) aberrant crypt foci (ACF) identified in HID-AB stained colon. All mutations in tumors and MDF caused amino acid substitution, while one mutation in ACF was silent. Beta-catenin detected at membrane level by immunohistochemistry was markedly reduced in MDF and tumors and, to a lesser extent, in ACF identified with HID-AB. By contrast, nuclear localization of beta catenin was significantly increased in MDF and tumors, while no variation was observed in ACF. Beta-catenin cytoplasmic expression was also significantly increased in MDF and tumors but to a lesser extent in ACF. In conclusion, MDF are induced dose-dependently by DMH, increase in size with time, have mutations in the beta-catenin gene and marked alterations in beta-catenin cellular localization. Since all these phenomena are considered specific steps for colon tumorigenesis, these results further support the hypothesis that MDF are cancer precursors and can be proposed as endpoints in short-term carcinogenesis experiments. PMID- 15751048 TI - Biliary lithiasis in early pregnancy and abnormal development of facial and distal limb bones (Binder syndrome): a possible role for vitamin K deficiency. AB - BACKGROUND: Binder syndrome is a maxillonasal dysostosis characterized by midface and nasal hypoplasia, sometimes associated with short terminal phalanges of fingers and toes and transient radiological features of chondrodysplasia punctata. Warfarin- or phenytoin-induced vitamin K deficiency during early pregnancy is a well-established etiology for this syndrome, which occurs nevertheless sporadically in most cases. CASE(S): We describe here the first case, to our knowledge, of Binder syndrome in a child whose mother presented with biliary lithiasis in early pregnancy. The mother proved to have a decrease in clotting factors II, VII, and X, and in prothrombin time, at 11 weeks of gestation, which was highly suggestive of vitamin K deficiency. CONCLUSIONS: The biliary lithiasis-induced vitamin K deficiency in early pregnancy is likely to have resulted in Binder syndrome. This observation should prompt physicians to carefully check for vitamin K deficiency in pregnant women presenting with biliary lithiasis, in order to prevent Binder syndrome in the fetus by providing intravenous vitamin K supplementation as soon as possible. Finally, reassuring genetic counseling regarding the genetic risk for future pregnancies is to be provided to the parents. PMID- 15751049 TI - Marked amelioration of alcohol-responsive posthypoxic myoclonus by gamma hydroxybutyric acid (Xyrem). AB - We conducted an open-label, dose-finding, blinded-rating trial of gamma hydroxybutyric acid (Xyrem) in a single patient with severe alcohol-responsive posthypoxic myoclonus refractory to treatment with standard antimyoclonic agents. Xyrem was given in divided doses during the day and was well tolerated. Intensity of myoclonus was measured using the Unified Myoclonus Rating Scale, and blinded videotape review demonstrated complete resolution of myoclonus at rest and stimulus-sensitive myoclonus. Action myoclonus and functional performance also improved in ways that were practically meaningful, allowing her to feed herself, to accomplish daily hygiene tasks, and to walk with assistance. The possible mechanisms of action and potential uses of this agent in other alcohol-responsive movement disorders are discussed. PMID- 15751050 TI - GP130/OSMR is the only LIF/IL-6 family receptor complex to promote osteoblast differentiation of calvaria progenitors. AB - Leukemia inhibitory factor (LIF) and its receptor (LIFR) are "twins" of Oncostatin M (OSM) and OSMR, respectively, likely having arisen through gene duplications. We compared their effects in a bone nodule-forming model of in vitro osteogenesis, rat calvaria (RC) cell cultures. Using a dominant-negative LIF mutant (hLIF-05), we showed that in RC cell cultures mouse OSM (mOSM) activates exclusively glycoprotein 130 (gp130)/OSMR. In treatments starting at early nodule formation stage, LIF, mOSM, IL-11, and IL-6 + sIL-6R inhibit bone nodule formation, that is, osteoprogenitor differentiation. Treatment with mOSM, and no other cytokine of the family, in early cultures (day 1-3 or 1-4) increases bone colony numbers. hLIF-05 also dose dependently stimulates bone nodule formation, confirming the inhibitory action of gp130/LIFR on osteogenesis. In pulse treatments at successive stages of bone nodule formation and maturation, LIF blocks osteocalcin (OCN) expression by differentiated osteoblasts, but has no effect on bone sialoprotein (BSP) expression. Mouse OSM inhibits OCN and BSP expression in preconfluent cultures with no or progressively reduced effects at later stages, reflecting the disruption of early nodules, possibly due to the strong apoptotic action of mOSM in RC cell cultures. In summary, LIFR and OSMR display differential effects on differentiation and phenotypic expression of osteogenic cells, most likely through different signal transduction pathways. In particular, gp130/OSMR is the only receptor complex of the family to stimulate osteoprogenitor differentiation in the RC cell culture model. PMID- 15751051 TI - Expression of the interferon-inducible chemokine IP-10 (CXCL10), a chemokine with proposed anti-neoplastic functions, in Hodgkin lymphoma and nasopharyngeal carcinoma. AB - Hodgkin lymphoma (HL) and nasopharyngeal carcinoma (NPC) are characterized by their association with Epstein-Barr virus (EBV) and an abundant infiltrate of reactive lymphoid cells. The presence of this lymphoid stroma may influence the effect of anti-viral immunotherapy. The interferon-inducible chemokine IP-10 has anti-neoplastic effects in several model systems mediated by T-cells expressing the CXCR3 chemokine receptor. Using in situ hybridization, it is shown that IP-10 is expressed in neoplastic cells of HL and correlates both with the mixed cellularity histotype and with EBV infection. IP-10 expression was also detected in tumour cells of most NPCs as well as in EBV-negative squamous cell carcinomas of the tongue. Thus, in carcinomas, IP-10 expression showed no correlation with EBV infection. Numerous CXCR3-positive lymphocytes were detected in the lymphoid stroma of HL and NPC, raising the possibility of a Th1-predominant immune response in these cases. In view of the proposed anti-neoplastic functions of IP 10 and CXCR3-positive lymphocytes, these findings are unexpected and raise the possibility that endogenous IP-10 expression in the context of human tumours may not exert the anti-tumour effects ascribed to it by in vitro experiments. PMID- 15751052 TI - A two-year prospective controlled study of bone mass and bone turnover in children with early juvenile idiopathic arthritis. AB - OBJECTIVE: To explore early changes and predictors of bone mass in children with juvenile idiopathic arthritis (JIA) in order to identify patients who will develop bone mass reductions. METHODS: We conducted a prospective cohort study of 108 children with early JIA (ages 6-18 years; mean disease duration 19.3 months) who were individually matched with 108 healthy children for age, sex, race, and county of residence. Bone mass and changes in total body, spine, femur, and forearm bone mineral density and bone mineral content (BMC), body composition, growth, and biochemical parameters of bone turnover were examined at baseline and at followup a mean of 24 months later. Low bone mass was defined as a Z score >1 SD below the reference population. RESULTS: Of the 200 children evaluated at followup, the 100 healthy children had greater gains in total body BMC (P = 0.035), distal radius BMC (P < 0.001), and total body lean mass (P < 0.001) than did the 100 JIA patients. Low or very low total body BMC was observed in 24% of the patients and 12% of the healthy children. Bone formation, bone resorption, and weight-bearing activities were reduced in the patients compared with the healthy children. Multiple regression analysis showed that in patients with JIA, serum bone-specific alkaline phosphatase, serum C-telopeptide of type I collagen, and weight-bearing activities were independent predictors of changes in total body BMC. Total body BMC was lower in patients with polyarticular onset than in those with oligoarticular disease onset. CONCLUSION: Patients with JIA have moderate reductions in bone mass gains, bone turnover, and total body lean mass early in the disease course. PMID- 15751053 TI - Cell turnover of serrated adenomas. AB - Serrated adenomas of the colon are characterized by epithelial neoplasia combining the architectural features of hyperplastic polyps and the cytological features of adenomas. Cell turnover, which is related to the malignant potential of these polyps, has not been thoroughly investigated. The aim of this study was to investigate epithelial cell proliferation, apoptosis, and oncoprotein expression in serrated adenomas. Twenty-five hyperplastic polyps, 25 serrated adenomas, and 25 tubulovillous adenomas resected from the colons of 75 patients were studied by immunohistochemical staining using monoclonal antibodies against MIB-1, Bcl-2, Bax, p53, and the TUNEL method for the detection of apoptosis. In serrated adenomas, the proliferation rate was significantly lower than in tubulovillous adenomas in both the lower and the upper parts of the crypts, and higher than that of hyperplastic polyps. Apoptosis was also significantly lower in serrated than in tubulovillous adenomas, but higher than in hyperplastic polyps. p53 oncoprotein expression was significantly greater in both serrated and tubulovillous adenomas than in hyperplastic polyps. bcl-2 protein expression was higher only in tubulovillous adenomas. Bax index was significantly different between tubullovillous and serrated adenomas, but the lowest values were observed in hyperplastic polyps. Serrated adenomas are highly proliferative polyps. They should be considered a biologically different entity from hyperplastic polyps. The intermediate features between serrated adenomas, hyperplastic polyps, and tubulovillous adenomas using the antibodies analysed in this study could have implications for the rate or the mechanism of development of malignancy in this type of polyp. PMID- 15751054 TI - Antiphospholipid syndrome nephropathy in antiphospholipid antibody-positive patients with systemic lupus erythematosus: comment on the article by Tektonidou et al. PMID- 15751056 TI - Classification analysis of the transcriptosome of nonlesional cultured dermal fibroblasts from systemic sclerosis patients with early disease. AB - OBJECTIVE: To compare the transcriptosome of early-passage nonlesional dermal fibroblasts from systemic sclerosis (SSc) patients with diffuse disease and matched normal controls in order to gain further understanding of the gene activation patterns that occur in early disease. METHODS: Total RNA was isolated from early-passage fibroblasts obtained from nonlesional skin biopsy specimens from 21 patients with diffuse SSc (disease duration <5 years in all but 1) and 18 healthy controls who were matched to the cases by age (+/-5 years), sex, and race. Array experiments were performed on a 16,659-oligonucleotide microarray utilizing a reference experimental design. Supervised methods were used to select differentially expressed genes. Quantitative polymerase chain reaction (PCR) was used to independently validate the array results. RESULTS: Of the 8,324 genes that passed filtering criteria, classification analysis revealed that <5% were differentially expressed between SSc and normal fibroblasts. Individually, differentially expressed genes included COL7A1, COL18A1 (endostatin), DAF, COMP, and VEGFB. Using the panel of genes discovered through classification analysis, a set of model predictors that achieved reasonably high predictive accuracy was developed. Analysis of 1,297 gene ontology (GO) classes revealed 35 classes that were significantly dysregulated in SSc fibroblasts. These GO classes included anchoring collagen (30934), extracellular matrix structural constituent (5201), and complement activation (6958, 6956). Validation by quantitative PCR demonstrated that 7 of 7 genes selected were concordant with the array results. CONCLUSION: Fibroblasts cultured from nonlesional skin of patients with SSc already have detectable abnormalities in a variety of genes and cellular processes, including those involved in extracellular matrix formation, fibrillogenesis, complement activation, and angiogenesis. PMID- 15751057 TI - Patients with antinuclear antibody-positive juvenile idiopathic arthritis constitute a homogeneous subgroup irrespective of the course of joint disease. AB - OBJECTIVE: We recently hypothesized that in the International League of Associations for Rheumatology (ILAR) classification of juvenile idiopathic arthritis (JIA), the presumably homogeneous patient group characterized by early onset of disease, a female predilection, the presence of antinuclear antibodies (ANA), asymmetric arthritis, and the risk for iridocyclitis is classified into different categories. We sought to investigate whether ANA-positive patients belonging to the ILAR categories of oligoarthritis and rheumatoid factor (RF) negative polyarthritis share homogeneous features and to compare these features with those of ANA-negative patients with JIA in the same categories. METHODS: We identified patients who were followed up during a 15-year period. All patients had JIA according to the ILAR criteria, with oligoarticular or polyarticular onset. ANA positivity was defined as 2 or more positive results at a titer of >or=1:160. Demographic and clinical features, including the number of joints involved over time and measures of JIA severity at the last followup visit, were recorded retrospectively. RESULTS: A total of 256 patients were included: 190 were ANA positive (109 had persistent oligoarthritis, 48 had extended oligoarthritis, and 33 had RF-negative polyarthritis), and 66 were ANA negative (35 had RF-negative polyarthritis, and 31 had oligoarthritis). All patients who were positive for ANA were similar in terms of age at disease presentation, female-to-male ratio, and frequency of symmetric arthritis and iridocyclitis. Compared with ANA-positive patients with polyarticular disease, ANA-negative patients with polyarticular arthritis were older at disease presentation and had a lower frequency of iridocyclitis, a higher frequency of symmetric arthritis, a greater cumulative number of joints affected over time, and a different pattern of joint disease, with a greater frequency of shoulder and hip involvement. The strong relationship between the presence of ANA and younger age at disease presentation, asymmetric arthritis, and development of iridocyclitis was confirmed by multivariate regression analysis. CONCLUSION: Our results support the hypothesis that patients with similar characteristics are currently classified into different JIA categories. The value of ANA positivity as a possible modifier of the current classification system deserves consideration. PMID- 15751058 TI - Induction of cartilage damage by overexpression of T cell interleukin-17A in experimental arthritis in mice deficient in interleukin-1. AB - OBJECTIVE: To examine the capacity of T cell interleukin-17A (IL-17A; referred to hereinafter as IL-17) to induce cartilage damage during experimental arthritis in the absence of IL-1. METHODS: Local IL-17 gene transfer was performed in the knee joint of IL-1-deficient mice and wild-type controls during streptococcal cell wall (SCW)-induced arthritis. Knee joints were isolated at various time points for histologic analysis of cartilage proteoglycan (PG) depletion. Expression of messenger RNA for inducible nitric oxide synthase, matrix metalloproteinases (MMPs) 3, 9, and 13, and ADAMTS-4 was determined by quantitative polymerase chain reaction analysis. VDIPEN staining was analyzed to study MMP-mediated cartilage damage. In addition, systemic anti-IL-1alpha/beta antibody treatment was performed in mice immunized with type II collagen and injected locally with an adenoviral vector expressing IL-17 or with control adenovirus. Knee joints were isolated and analyzed for cartilage PG depletion, chondrocyte death, and cartilage surface erosion. RESULTS: During SCW-induced arthritis, local T cell IL 17 gene transfer turned this acute, macrophage-driven joint inflammation into a severe, chronic arthritis accompanied by aggravated cartilage damage. Of high interest, the IL-1 dependency of cartilage PG depletion was fully abrogated when IL-17 was locally overexpressed in the joint. Moreover, local IL-17 gene transfer increased MMP expression without the need for IL-1, although IL-1 remained essential for part of the cartilage VDIPEN expression. Furthermore, when IL-17 was overexpressed in the knee joints of mice with collagen-induced arthritis, anti-IL-1 treatment did not reduce the degree of chondrocyte death or cartilage surface erosion. CONCLUSION: These data show the capacity of IL-17 to replace the catabolic function of IL-1 in cartilage damage during experimental arthritis. PMID- 15751059 TI - Academic rheumatology: not just a man's world. PMID- 15751060 TI - HLA-B27-restricted CD8+ T cell response to cartilage-derived self peptides in ankylosing spondylitis. AB - OBJECTIVE: Several hypotheses have been proposed to explain the strong association between HLA-B27 and ankylosing spondylitis (AS). Among these, the arthritogenic peptide theory proposes that certain B27 subtype alleles bind specific arthritogenic peptide(s) due to their unique amino acid anchor residues. Cartilage antigens have been discussed as candidate targets for the immune response in AS. The recognition of HLA-B27-peptide complexes by self-reactive CD8+ T cells might contribute to joint-specific tissue damage. Therefore, we investigated the presence of autoreactive CD8+ T cells specific for cartilage derived peptides in patients with AS. METHODS: An HLA-B27-binding prediction program and a proteasome-cutting prediction program for the human 20S proteasome were used to screen 18 human cartilage proteins for potentially immunogenic nonamer peptides. The peptides identified were used to stimulate peripheral blood mononuclear cells from 20 HLA-B27-positive patients with AS and synovial fluid (SF) mononuclear cells from 7 HLA-B27-positive patients with AS. Activation of T cells was measured by antigen-specific intracellular cytokine staining and quantified by flow cytometry. RESULTS: From the screening analysis, we identified 121 nonamer peptides. Of these, 1 peptide derived from type II collagen and 1 from type VI collagen were stimulatory for peripheral blood CD8+ T cells in only 1 of 20 patients. However, in 4 of 7 SF samples the same type VI collagen-derived nonamer peptide stimulated SF CD8+ T cells, but none of the other peptides was stimulatory. This CD8+ T cell response could be blocked by an anti-HLA-B27 antibody, confirming an HLA-B27-restricted immune response. CONCLUSION: Our findings suggest that cartilage-directed cellular autoimmunity might play an important role in joint-specific tissue damage in patients with AS. Future research is necessary to determine whether the identified peptide is of pathogenetic relevance. PMID- 15751061 TI - Joint inflammation and chondrocyte death become independent of Fcgamma receptor type III by local overexpression of interferon-gamma during immune complex mediated arthritis. AB - OBJECTIVE: It has previously been shown that the onset and the degree of joint inflammation during immune complex (IC)-mediated arthritis depend on Fcgamma receptor type III (FcgammaRIII). Local adenoviral overexpression of interferon gamma (IFNgamma) in the knee joint prior to onset of IC-mediated arthritis aggravated severe cartilage destruction. In FcgammaRI(-/-) mice, however, chondrocyte death was not enhanced by IFNgamma, whereas matrix metalloproteinase (MMP)-mediated aggrecan breakdown was markedly elevated, suggesting a role for the activating FcgammaRIII in the latter process. We undertook this study to determine the role of FcgammaRIII in joint inflammation and severe cartilage destruction in IFNgamma-stimulated IC-mediated arthritis, using FcgammaRIII(-/-) mice. METHODS: FcgammaRIII(-/-) and wild-type (WT) mice were injected in the knee joint with recombinant adenovirus encoding murine IFNgamma (AdIFNgamma) or with adenovirus encoding enhanced green fluorescent protein 1 day prior to induction of IC-mediated arthritis. Histologic sections were obtained 3 days after arthritis onset to study inflammation and cartilage damage. MMP-mediated expression of the VDIPEN neoepitope was detected by immunolocalization. Chemokine and FcgammaR expression levels were determined in synovial washouts and synovium, respectively. RESULTS: Injection of AdIFNgamma in naive knee joints markedly increased levels of messenger RNA for FcgammaRI, FcgammaRII, and FcgammaRIII. Upon IFNgamma overexpression prior to induction of IC-mediated arthritis, joint inflammation was similar in FcgammaRIII(-/-) and WT mice. The percentage of macrophages in the knee joint was increased, which correlated with high concentrations of the macrophage attractant macrophage inflammatory protein 1alpha. Furthermore, IFNgamma induced 2-fold and 3-fold increases in chondrocyte death in WT controls and FcgammaRIII(-/-) mice, respectively. Notably, VDIPEN expression also remained high in FcgammaRIII(-/-) mice. CONCLUSION: IFNgamma bypasses the dependence on FcgammaRIII in the development of IC-mediated arthritis. Furthermore, both FcgammaRI and FcgammaRIII can mediate MMP-dependent cartilage matrix destruction. PMID- 15751062 TI - A novel ultrasonographic synovitis scoring system suitable for analyzing finger joint inflammation in rheumatoid arthritis. AB - OBJECTIVE: To develop an ultrasonographic (US) synovitis scoring system suitable for evaluation of finger joint inflammation in patients with active rheumatoid arthritis (RA) and to compare semiquantitative US scoring with quantitative US measurements. METHODS: US was performed at the palmar and dorsal sides of the second through fifth metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints in 10 healthy subjects and in the clinically more affected hand in 46 RA patients. Ten patients additionally underwent magnetic resonance imaging (MRI). Synovitis was measured, standardized, and scored according to a semiquantitative method. The 2 methods (semiquantitative US scoring, quantitative US) were compared and statistical cutoffs were identified using receiver operating characteristic (ROC) curve analysis. MRI results were compared with semiquantitative US scoring and quantitative US results. The optimal US scoring method from 6 joint combinations was identified (ROC curve analysis). RESULTS: Synovitis was most frequently detected in the palmar proximal area (86% of affected joints). We found no significant differences between individual PIP joints or between individual MCP joints, indicating that all fingers within each of these joint groups should be treated equally for statistical calculations, although each joint group as a whole should be treated separately. The optimal cutoff point to distinguish between "health" and "pathology" was 0.6 mm both for MCP joints (sensitivity 94%, specificity 89%) and for PIP joints (sensitivity 90%, specificity 88%). There was no significant difference between semiquantitative US scores and quantitative US measurements. The best results for joint combinations were achieved using the "sum of 4 fingers" (second through fifth MCP and PIP joints) and "sum of 3 fingers" (second through fourth MCP and PIP joints) methods. Comparison of MRI results with semiquantitative US scores revealed high concordance. CONCLUSION: US evaluation of finger joint synovitis can be considerably simplified by focusing on the palmar side and by applying semiquantitative grading instead of quantitative measurements. For evaluation of treatment efficacy based on synovitis in RA patients, we recommend using the "sum of 3 fingers" method in longitudinal trials. PMID- 15751063 TI - Electroencephalography in the assessment of neuropsychiatric manifestations in antiphospholipid syndrome and systemic lupus erythematosus. AB - OBJECTIVE: To describe the prevalence and clinical associations of abnormalities on electroencephalography (EEG) in patients with antiphospholipid syndrome (APS) and/or systemic lupus erythematosus (SLE) who have neuropsychiatric symptoms. METHODS: The study group comprised 57 patients (age or=55 years) from the Rotterdam Study. Subjects with a baseline Kellgren/Lawrence grade of 0 or 1 in both hips were included in the study. Incident radiographic OA of the hip was defined as a decrease of joint space width of the hip (>or=1.0 mm) at followup. Acetabular dysplasia was assessed using the center-edge angle and the acetabular depth. The association between acetabular dysplasia and incident radiographic hip OA was assessed by calculating odds ratios using multivariate regression analysis. RESULTS: In this study population with a mean +/- SD age of 65.6 +/- 6.5 years, 9.3% developed incident radiographic hip OA. Subjects with acetabular dysplasia (center-edge angle <25 degrees ) had a 4.3-fold increased risk for incident radiographic OA of the hip (95% confidence interval 2.2-8.7) compared with subjects without acetabular dysplasia. These associations were independent of known determinants of hip OA such as age, sex, and body mass index (BMI), but tended to be enhanced by female sex, heavy mechanical load, and low BMI. CONCLUSION: In a study population age >or=55 years, acetabular dysplasia is still a strong independent determinant of incident radiographic hip OA. PMID- 15751072 TI - Investigation of the SLC22A4 gene (associated with rheumatoid arthritis in a Japanese population) in a United Kingdom population of rheumatoid arthritis patients. AB - OBJECTIVE: Recent studies of 2 complex diseases, rheumatoid arthritis (RA) and Crohn's disease (CD), showed associations with genes mapping to the cytokine gene cluster on 5q31. In particular, a functional single-nucleotide polymorphism (SNP) mapping to intron 1 of the organic cation transporter 1 (OCTN1; SLC22A4) gene was associated with RA in a Japanese population, and a haplotype of a different SNP in the same gene and one in an adjacent gene, OCTN2 (SLC22A5), was associated with CD. The purpose of this study was to investigate the association between the OCTN locus and RA in a Caucasian population. METHODS: Association with 11 SNPs spanning the SLC22A4 and SLC22A5 genes, including a putative RA-causing functional polymorphism (rs3792876 [slc2f2]) and a functional haplotype previously associated with CD, was investigated in 909 RA cases and 594 population controls in the UK. Genotyping was performed using 5'-allele discrimination assays. Estimated haplotype frequencies were generated using the expectation-maximization algorithm and were compared between cases and controls. RESULTS: All SNPs were in Hardy-Weinberg equilibrium. We found no evidence for an association between RA and either the SNP (rs3792876 [slc2f2]) or the haplotype previously reported to be associated with RA in a Japanese population. Similarly, no association between RA and the haplotype associated with CD was detected. CONCLUSION: Functional polymorphisms of the OCTN gene locus that have previously been associated with RA and CD were not found to be associated with RA in a UK population. The findings do not provide support for a major role of these genes in the etiology of RA in this population. PMID- 15751073 TI - Rosiglitazone induces interleukin-1 receptor antagonist in interleukin-1beta stimulated rat synovial fibroblasts via a peroxisome proliferator-activated receptor beta/delta-dependent mechanism. AB - OBJECTIVE: To study the potency of 2 peroxisome proliferator-activated receptor gamma (PPARgamma) agonists, 15-deoxy-Delta(12,14)-prostaglandin J(2) (15-deoxy PGJ(2)) and rosiglitazone, to modulate the expression of interleukin-1 receptor antagonist (IL-1Ra) in rat synovial fibroblasts. METHODS: Levels of messenger RNA for IL-1Ra and PPAR isotypes (alpha, beta/delta, gamma) were assessed by real time polymerase chain reaction in rat synovial fibroblasts exposed to 10 ng/ml of IL-1beta. PPAR levels were assessed by Western blotting and secreted IL-1Ra levels by immunoassay. The potency of PPARgamma agonists and the PPARbeta/delta agonist GW-501516 on IL-1Ra levels was tested in the range of 1-10 microM and at 100 pM, respectively. The contribution of PPARgamma to the effects of rosiglitazone on IL-1Ra secretion was examined either by its overexpression or by inhibition using wild-type or dominant-negative constructs and the antagonist GW 9662 (10 microM), respectively. The dominant-negative strategy was also performed to investigate the possible contribution of PPARbeta/delta and NF-kappaB activation. RESULTS: IL-1beta-induced IL-1Ra production was increased by 10 microM rosiglitazone but was reduced dose-dependently by 15-deoxy-PGJ(2). Both agonists lowered IL-1beta secretion, but rosiglitazone alone reduced the imbalance of IL-1beta/IL-1Ra toward basal levels. Enhancement of IL-1beta-induced IL-1Ra production by rosiglitazone was not affected by PPARgamma overexpression or by its inhibition with dominant-negative PPARgamma or GW-9662. Inhibition of NF-kappaB was also ineffective against rosiglitazone but abolished the stimulating effect of IL-1beta on IL-1Ra. All PPAR isotypes were expressed constitutively in rat synoviocytes, but PPARgamma decreased dramatically upon IL 1beta exposure, whereas PPARbeta/delta remained stable. Dominant-negative PPARbeta/delta abolished the enhancement of IL-1Ra by rosiglitazone, whereas GW 501516 reproduced the effect of rosiglitazone on IL-1Ra secretion. CONCLUSION: Rosiglitazone stimulates IL-1Ra production by a PPARbeta/delta mechanism in activated rat synovial fibroblasts, further contributing to its potential antiarthritic properties and opening new perspectives for the modulation of inflammatory genes by specific PPAR agonists in articular cells. PMID- 15751074 TI - Chemokines and their receptors in rheumatoid arthritis: future targets? PMID- 15751076 TI - Septic arthritis due to Listeria monocytogenes in a patient receiving etanercept. PMID- 15751075 TI - Magnetic resonance imaging evidence of tendinopathy in early rheumatoid arthritis predicts tendon rupture at six years. AB - OBJECTIVE: To determine whether magnetic resonance imaging (MRI) evidence of tendinopathy in early rheumatoid arthritis (RA) could be used to predict the course of tendon involvement in later disease and specifically the risk of tendon rupture. METHODS: The occurrence, pattern, and progression of tendinopathy were studied prospectively over 6 years in a cohort of patients who had presented with RA. Of 42 patients enrolled, full MRI and clinical data were available for 31 at 6 years. MRI scans of the dominant wrist were scored for tendinopathy by 2 radiologists using a validated system. These data were compared with MRI synovitis, erosion scores, and disease activity measures. Prognostic factors for tendon inflammation and rupture were sought. RESULTS: Thirty-four patients (81%) had MRI evidence of tendinopathy at baseline, falling to 59% at 1 year and 68% at 6 years. The most commonly affected site was the extensor carpi ulnaris. MRI tendinopathy and synovitis scores were correlated at baseline (r = 0.37, P = 0.01) and 1 year (r = 0.45, P = 0.003) but not at 6 years (r = 0.11, P = 0.5). The strongest predictor of the 6-year tendinopathy score was the 1-year tendinopathy score (R(2) = 0.36, P = 0.0003 [beta = 1.28, SE = 0.31]). In 4 patients, extensor tendon rupture had occurred by 6 years. Their baseline and 1 year tendinopathy scores were higher than in the nonrupture group (medians 2.8 versus 1.0 [P = 0.04] and 4.3 versus 0.8 [P = 0.03], respectively), as were their Health Assessment Questionnaire scores (1.33 versus 0.54 [P = 0.02], 1.18 versus 0.25 [P = 0.03], and 0.98 versus 0.37 [P = 0.01] at 0, 1, and 6 years, respectively). For the group as a whole, the baseline tendinopathy score was predictive of rupture at 6 years (odds ratio [OR] 1.52, 95% confidence interval [95% CI] 1.02-2.32, P = 0.03), as was the 1-year score (OR 1.57, 95% CI 1.03 2.04, P = 0.02). CONCLUSION: MRI can be used to quantify tendinopathy at the wrist in RA patients. High scores in early disease were predictive of tendon rupture in a small group of patients, but further studies are required to determine whether this has clinical relevance. PMID- 15751077 TI - Expression of interleukin-21 receptor in epidermis from patients with systemic sclerosis. AB - OBJECTIVE: To examine the expression and regulation of interleukin-21 (IL-21) and IL-21 receptor (IL-21R) in patients with systemic sclerosis (SSc; scleroderma). METHODS: Skin biopsy specimens were obtained from 23 patients with SSc and 15 healthy controls. IL-21/IL-21R messenger RNA (mRNA) was quantified using real time polymerase chain reaction (PCR). The expression pattern of IL-21/IL-21R was analyzed by in situ hybridization and Western blotting. Stimulation experiments were performed with cultured dermal fibroblasts from patients with SSc and healthy controls as well as with keratinocytes, using IL-1beta, platelet-derived growth factor BB, monocyte chemoattractant protein 1, transforming growth factor beta, and IL-21. The SCID-hu skin mouse model was used for in vivo experiments. RESULTS: IL-21R mRNA was detected in all biopsy specimens from patients with SSc and controls, with a 4.7-fold increase observed in SSc samples. In situ hybridization and immunohistochemical analysis showed an up-regulation of IL-21R in samples of epidermis from SSc patients, whereas no signal was detected in skin specimens from healthy controls. These results were confirmed in vitro, in that cultured keratinocytes expressed significant levels of IL-21R, whereas no signal was observed in fibroblasts. Interestingly, mRNA for IL-21 could not be detected by real-time PCR and in situ hybridization. Various concentrations of key cytokines in the pathogenesis of SSc did not stimulate the expression of IL-21R mRNA in cultured keratinocytes and dermal fibroblasts. In the SCID mouse transplantation model, the overexpression of IL-21R mRNA in SSc keratinocytes remained unchanged after transplantation. CONCLUSION: The up-regulation of IL-21R in keratinocytes indicates that, similar to fibroblasts and endothelial cells, the expression pattern is altered in SSc. Moreover, the expression of IL-21R appears to be independent of key cytokines that are operant in SSc. PMID- 15751078 TI - Glucocorticoids inhibit osteocalcin transcription in osteoblasts by suppressing Egr2/Krox20-binding enhancer. AB - OBJECTIVE: Glucocorticoids are widely used for the management of rheumatoid arthritis. Osteoporosis is a major side effect of glucocorticoid therapy and is attributable to inhibition of bone formation. We developed an osteoblast culture system in which glucocorticoids strongly inhibit development of the osteoblast phenotype, including expression of the bone-specific osteocalcin (OC) gene. Using this gene as a model, the goal of this study was to discover glucocorticoid sensitive transcriptional mechanisms in osteoblasts. METHODS: Dexamethasone (DEX; 1 microM) was administered to murine MC3T3-E1 osteoblastic cultures under conditions that inhibit mineralized extracellular matrix formation and OC messenger RNA levels by >10-fold. Because standard (short-term) transient transfection assays with OC promoter-reporter constructs did not recapitulate the strong DEX-mediated repression, mapping of OC negative glucocorticoid response elements (GREs) was performed initially by stable transfection and then with long term transient transfection assays. Transcription factor binding to the OC negative GRE was studied by electrophoretic mobility shift assays. RESULTS: Several-fold repression of OC-luciferase constructs was recapitulated in stable and long-term transient transfection assays, in which the transfected cells were allowed to progress to a sufficiently advanced developmental stage. Analysis of a 5' promoter deletion series mapped an OC negative GRE to a 15-bp G/C-rich motif ( 161/-147) located just upstream of the binding site for the osteoblast master transcription factor Runx2. Oligonucleotides encompassing this element and MC3T3 E1 cell extracts formed a protein-DNA complex that contained an Egr/Krox family member(s). Complex formation was competed by either an oligonucleotide containing 2 consensus Egr motifs or by anti-Egr2/Krox20 antibodies. Three copies of this Krox-binding element conferred 20-fold transcriptional activation on the 147-bp basal OC promoter in osteoblasts, and the enhancer activity was inhibited by DEX. Enhancer activity was not observed in 10T1/2 fibroblasts unless these cells were cotransfected with Runx2. CONCLUSION: An Egr2/Krox20-binding site located immediately upstream of the Runx2 site of the mouse OC promoter was identified as an enhancer in osteoblasts, whose activity is repressed by glucocorticoids. Sequence similarity suggests that such a mechanism is likely operative in both murine and human cells. Because glucocorticoids inhibit Egr2/Krox20 expression in osteoblasts, and because trabecular bone formation is arrested in Egr2/Krox20 knockout mice, the inhibition of Egr2/Krox20 activity likely contributes to glucocorticoid-induced osteoporosis. PMID- 15751079 TI - Amelioration of acute inflammation by systemic administration of a cell-permeable peptide inhibitor of NF-kappaB activation. AB - OBJECTIVE: We used an experimental model of inflammation in mice, carrageenan induced paw edema, to study the antiinflammatory effects of the NEMO-binding domain (NBD) peptide, which blocks activation of the inducible transcription factor NF-kappaB. METHODS: Paw edema was induced by subplantar injection of 1% lambda-carrageenan into the mouse left hind paw. Test agents were given intraperitoneally immediately after carrageenan injection. The increase in footpad thickness was considered to be edema. In some experiments, the mice were killed and the paws were removed for histologic and molecular biology analysis. NF-kappaB DNA binding activity was evaluated in nuclear extracts by electrophoretic mobility shift assays. The expression levels of NF-kappaB regulated cyclooxygenase 2 (COX-2) protein and tumor necrosis factor alpha (TNFalpha) messenger RNA (mRNA) were evaluated by immunoblot analysis and polymerase chain reaction amplification of reverse-transcribed mRNA, respectively. RESULTS: We found that systemically administered NBD peptide significantly inhibited edema formation and cellular infiltration in inflamed mouse paws. This antiinflammatory activity was most likely due to inhibition of expression of proinflammatory mediators, such as TNFalpha and COX-2, in inflamed tissues. CONCLUSION: These studies further establish NF-kappaB as a target for antiinflammatory therapy and provide support for the use of the NBD peptide as a possible therapeutic agent for inflammatory diseases. PMID- 15751080 TI - Hyaluronan oligosaccharide-induced activation of transcription factors in bovine articular chondrocytes. AB - OBJECTIVE: To document the activity profile of transcription factors following chondrocyte stimulation with hyaluronan (HA) hexasaccharides (HA(6)) and to determine the expression of genes whose transcriptional activation is tightly associated with the transcription factors. METHODS: Nuclear extracts from bovine articular chondrocytes treated with HA(6) were subjected to transcription factor protein-DNA array analysis. Electrophoretic mobility shift assay (EMSA) analyses were performed to confirm the results of protein-DNA array. The gene expressions of matrix metalloproteinase 3 (MMP-3), type II collagen, and cartilage oligomeric matrix protein (COMP) were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), and protease activity was assessed by casein zymography. RESULTS: In the protein-DNA array analysis, 12 transcription factors were up-regulated and 2 transcription factors were down regulated in the chondrocytes treated with HA(6). The transcription factors retinoic acid receptor (RAR), retinoid X receptor (RXR), and Sp-1 exhibited >2 fold increased activity by HA(6) treatment, as confirmed by EMSA. RT-PCR analysis showed that the expression levels of MMP-3, type II collagen, and COMP messenger RNA, which are tightly associated with the activation of RAR, RXR, or Sp-1, were up-regulated by treatment with HA(6). Addition of high molecular mass HA after HA(6) treatment resulted in abrogation of the MMP-3 induction. CONCLUSION: These results suggest that HA(6) increase the activity of multiple transcription factors in chondrocytes and signal the enhanced expression of key genes involved in cartilage-matrix remodeling and turnover. The data also demonstrate that high molecular mass HA has a potential to suppress the signaling activated by HA(6). PMID- 15751082 TI - Targeting the interleukin-6 receptor: a new treatment for systemic juvenile idiopathic arthritis? PMID- 15751081 TI - Anti-ribosomal P protein antibody in human systemic lupus erythematosus up regulates the expression of proinflammatory cytokines by human peripheral blood monocytes. AB - OBJECTIVE: Autoantibodies to ribosomal P proteins (anti-P antibodies) are detected in 12-16% of patients with systemic lupus erythematosus (SLE) and have been found to be associated with some manifestations of the disease, including lupus psychosis and hepatitis. Recent studies have disclosed that anti-P antibodies react with activated T cells but not with B cells, suggesting possible direct effects of anti-P antibodies on immune regulation. The present study was designed to explore the presence of the epitope recognized by anti-P antibodies on human peripheral blood monocytes. METHODS: Highly purified peripheral blood monocytes obtained from healthy donors were cultured with or without interferon gamma (IFNgamma) in the presence of either anti-P antibodies purified by affinity chromatography from the sera of patients with SLE or control IgG. RESULTS: Flow cytometry analysis disclosed that fresh (day 0) monocytes did not express the ribosomal P epitope, whereas expression of the ribosomal P epitope was induced on annexin V-negative monocytes after activation through plastic adherence for 48 hours. More important, anti-P antibodies (compared with normal IgG or IgG from SLE patients devoid of anti-P antibodies) enhanced the production of tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) by activated monocytes. Accordingly, anti-P antibodies also up-regulated the expression of TNFalpha and IL-6 messenger RNA in activated monocytes. Of note, F(ab')(2) fragments of anti-P antibodies, which do not result in Fcgamma receptor (FcgammaR) crosslinking, also effectively up-regulated the expression of TNFalpha and IL-6. CONCLUSION: These results indicate that human peripheral blood monocytes express the ribosomal P epitope upon activation, irrespective of induction of apoptosis. Moreover, the data suggest that anti-P antibodies might modify a variety of inflammatory responses through up-regulation of the expression of proinflammatory cytokines in monocytes, in a manner that does not involve FcgammaR crosslinking. PMID- 15751083 TI - The national bone and joint decade: impact for rheumatic diseases. PMID- 15751084 TI - Accelerating effect of an MRL gene locus on the severity and onset of arthropathy in DBA/1 mice. AB - OBJECTIVE: To analyze the influence of the genetic background of an arthritis prone strain of mice, MRL, on the spontaneous development of arthropathy in DBA/1 mice, which histopathologically resembles enthesopathy in humans, and to clarify the strain-specific gene loci and their interactions that confer susceptibility to arthropathy. METHODS: MRL, DBA/1, (MRL x DBA/1)F(1), and (MRL x DBA/1)F(2) intercross mice were prepared, and the severity and onset of arthropathy of the ankle joints in individual mice were quantified (0-3 and 0-5 scale, respectively). A genome-wide scan of 271 male F(2) intercross mice with polymorphic microsatellite markers was performed. RESULTS: Only male DBA/1, (MRL x DBA/1)F(1), and (MRL x DBA/1)F(2) mice developed arthropathy. The macroscopic and histopathologic findings of arthropathy in the F(2) mice were similar to those in the parental DBA/1 mice, but the onset was significantly earlier. In the quantitative trait locus analysis of male F(2) mice, 1 susceptibility locus for both the severity and early onset of the disease in the region of an MRL allele, Amd1, was located at marker D10Mit259 (map position 40.0 cM), which was common to 1 of the sialadenitis susceptibility loci in MRL mice, Asm1. Another susceptibility locus for the severity and early onset of arthropathy in the region of a DBA allele, Amd2, was located at D3Mit46 (29.5 cM). These loci manifested an additive effect on the development of arthropathy. CONCLUSION: Arthropathy in DBA/1 mice is under the control of an allelic combination of gene loci, one of which is common to the locus for sialadenitis in MRL/MpJ-lpr/lpr mice. PMID- 15751087 TI - Clinical images: Pachydermodactyly. PMID- 15751086 TI - Erythropoietin reduces the degree of arthritis caused by type II collagen in the mouse. AB - OBJECTIVE: Erythropoietin (EPO) is a potent stimulator of erythroid progenitor cells, and its expression is enhanced by hypoxia. The aim of this study was to investigate the effect of EPO on collagen-induced arthritis (CIA) in the mouse. METHODS: CIA was induced by intradermal injection of bovine type II collagen (CII) and Freund's complete adjuvant. Starting on day 25, some of the mice with CIA received daily subcutaneous injections of EPO (1,000 units/kg). Two other groups of mice received sham treatment alone or sham treatment followed by EPO treatment, respectively. Arthritis was assessed clinically, radiologically, and histologically. Cytokine and chemokine levels were measured, and neutrophil infiltration into inflamed joints was quantitated. Immunohistochemistry studies were performed to measure protein nitrosylation. Chondrocyte apoptosis was assessed by TUNEL assay. RESULTS: Macroscopic clinical evidence of CIA first appeared as periarticular erythema and edema in the hind paws. The incidence of CIA was 100% by day 27 in the CII-challenged mice, and the severity of CIA progressed over a 35-day period, with radiographic evaluation revealing focal resorption of bone. Histopathologic features of CIA included erosion of the cartilage at the joint margins. Treatment with EPO starting at the onset of arthritis (day 25) ameliorated the clinical signs on days 26-35 and improved histologic status in the joints and paws. The degree of oxidative and nitrosative damage was significantly reduced in EPO-treated mice as indicated by decreased nitrotyrosine formation and poly(ADP-ribose) polymerase activation. Plasma levels of the proinflammatory cytokine tumor necrosis factor alpha were also significantly reduced by EPO treatment. In addition, EPO reduced the levels of apoptosis in chondrocytes in articular cartilage, as indicated by decreased TUNEL staining. CONCLUSION: These findings demonstrate that EPO exerts an antiinflammatory effect during chronic inflammation and is able to ameliorate the tissue damage associated with CIA. PMID- 15751088 TI - Increase in the expression of the transmembrane surface receptor CD44v6 on chondrocytes in animals with osteoarthritis. AB - OBJECTIVE: To evaluate the expression of CD44v6 on chondrocytes from hyaline cartilage over the course of osteoarthritis (OA). METHODS: In 12 NZW rabbits, the anterior cruciate ligament (ACL) was resected to create anterior instability of the knee. In 12 control rabbits, only a sham operation, without resection of the ACL, was performed. Four animals from each group were killed at 3, 6, and 12 weeks. After opening the knee joint, OA was macroscopically graded and hyaline cartilage of the load-bearing area was evaluated histologically according to the Mankin scale and by immunostaining for CD44v6. RESULTS: There was a positive linear correlation between the time after surgery and the macroscopic and histologic OA scores. The scores in the control group were constant over the time course. Immunostaining showed constant expression of CD44v6 in the control group. In the experimental group, a positive linear correlation between CD44v6 expression and macroscopic and histologic grades was found. CONCLUSION: The results show an in vivo increase in the expression of the hyaluronan receptor CD44 over the time course of OA. Further studies are needed to evaluate whether this pattern applies to humans and whether new treatment approaches could evolve from this knowledge. PMID- 15751089 TI - Evaluation of the pituitary-adrenal axis function following single intraarticular injection of methylprednisolone. AB - OBJECTIVE: To determine the effects of a single session of intraarticular methylprednisolone (IA-MP) injections on the function of the pituitary-adrenal axis. METHODS: Twenty-five patients with rheumatic diseases were treated with a single IA-MP injection. The dose varied between 20 and 160 mg. The basal cortisol level and the response to 1 microg adrenocorticotropic hormone (ACTH) stimulation were determined before treatment, at 1 day, and at 1 week after IA-MP injections. Further tests were carried out for up to 6 weeks in patients with blunted cortisol response. Results were compared with those obtained in a group of 22 healthy subjects. RESULTS: Before treatment, all patients had normal basal and peak cortisol responses to ACTH (>396 nmoles/liter at 30 minutes). Reduced fasting cortisol levels (<147 nmoles/liter) were detected in 12 of 25 patients (48%) after 1 day, in 1 of 25 patients (4%) after 1 week, and in 1 of 25 patients (4%) 2 weeks after IA-MP injections. Blunted peak cortisol response (<396 nmoles/liter at 30 minutes) was observed in 1 of 25 patients (4%) after 1 day, in 3 of 25 patients (12%) after 1 week, and in 1 of 25 patients (4%) 2 weeks after IA-MP injections. Decreased fasting levels and peak cortisol responses to ACTH stimulation were more common in patients with inflammatory diseases and in those injected with 80-160 mg MP. CONCLUSION: Single-session IA-MP injection(s) are associated with systemic absorption of MP, causing impaired adrenocortical reserve. Recovery is expected in most patients after 1-2 weeks. Only a few patients exhibited suppression for up to 2 weeks. The magnitude of this suppression depends on the dose injected, and is more common in patients with inflammatory joint diseases. Caution is required if repeated large doses of IA-MP are considered in these patients. PMID- 15751090 TI - Febuxostat, a novel nonpurine selective inhibitor of xanthine oxidase: a twenty eight-day, multicenter, phase II, randomized, double-blind, placebo-controlled, dose-response clinical trial examining safety and efficacy in patients with gout. AB - OBJECTIVE: Gout affects approximately 1-2% of the American population. Current options for treating hyperuricemia in chronic gout are limited. The purpose of this study was to assess the safety and efficacy of febuxostat, a nonpurine selective inhibitor of xanthine oxidase, in establishing normal serum urate (sUA) concentrations in gout patients with hyperuricemia (>or=8.0 mg/dl). METHODS: We conducted a phase II, randomized, double-blind, placebo-controlled trial in 153 patients (ages 23-80 years). Subjects received febuxostat (40 mg, 80 mg, 120 mg) or placebo once daily for 28 days and colchicine prophylaxis for 14 days prior to and 14 days after randomization. The primary end point was the proportion of subjects with sUA levels <6.0 mg/dl on day 28. RESULTS: Greater proportions of febuxostat-treated patients than placebo-treated patients achieved an sUA level <6.0 mg/dl at each visit (P < 0.001 for each comparison). The targeted sUA level was attained on day 28 in 0% of those taking placebo and in 56% of those taking 40 mg, 76% taking 80 mg, and 94% taking 120 mg of febuxostat. The mean sUA reduction from baseline to day 28 was 2% in the placebo group and 37% in the 40 mg, 44% in the 80-mg, and 59% in the 120-mg febuxostat groups. Gout flares occurred with similar frequency in the placebo (37%) and 40-mg febuxostat (35%) groups and with increased frequency in the higher dosage febuxostat groups (43% taking 80 mg; 55% taking 120 mg). During colchicine prophylaxis, gout flares occurred less frequently (8-13%). Incidences of treatment-related adverse events were similar in the febuxostat and placebo groups. CONCLUSION: Treatment with febuxostat resulted in a significant reduction of sUA levels at all dosages. Febuxostat therapy was safe and well tolerated. PMID- 15751091 TI - A serum proteomic approach to gauging the state of remission in Wegener's granulomatosis. AB - OBJECTIVE: To identify serum ion patterns that distinguish remission from active disease in patients with Wegener's granulomatosis (WG). METHODS: Using sera collected in the WG Etanercept Trial, we selected samples from patients who either were undergoing a period of extended disease remission or had recent flares of active WG. Unfractionated samples were randomized into sets for training and testing, such that remission sera and active disease sera could be analyzed without batch bias. Molecular species within the sera were ionized by high-resolution, matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We then used a bioinformatics pattern-recognition tool to identify optimal combinations of ions. During the training stage, the clinical data (remission versus active disease) were provided in association with the spectral data from each sample. In the testing stage, we performed blinded testing on a previously unexamined set of samples. RESULTS: The most robust model, trained on a total of 82 samples (42 remission, 40 active disease), included 7 key ions with mass:charge ratios of 803.239, 2,171.672, 2,790.574, 3,085.237, 5,051.726, 5,833.989, and 6,630.465. The combined relative amplitudes of these 7 ions identified 5 distinct clusters of either remission or active disease samples during the training stage. In the testing stage, this model segregated 72 samples into the same 5 clusters, including 1 large remission cluster (n = 28) and another large active disease cluster (n = 32). Three smaller clusters of active disease or remission samples were also identified, with remission clusters populated by 2 samples in one cluster and 8 in another, and an active disease cluster populated by 2 samples. The model categorized 35 of 37 remission samples correctly (sensitivity 95%, 95% confidence interval [95% CI] 82.1-99.4) and 32 of 35 active disease samples correctly (specificity 91%, 95% CI 78.1-98.1). CONCLUSION: This serum proteomic profiling approach appears to be useful in distinguishing between states of stable clinical remission and active disease. Further validation and refinement of this strategy may help clinicians apply immunosuppressive therapies more judiciously among their patients, thereby avoiding morbidity and mortality from excessive treatment. Identification of the most robust and clinically useful combinations of ions will permit the rational selection of molecules for sequencing and analysis. PMID- 15751092 TI - Recruitment of topoisomerase I (Scl-70) to nucleoplasmic proteasomes in response to xenobiotics suggests a role for altered antigen processing in scleroderma. AB - OBJECTIVE: Scleroderma, also known as systemic sclerosis (SSc), is a chronic, life-threatening autoimmune disease characterized by a wide spectrum of manifestations and a variable evolution. The presence of particular antinuclear antibodies is often predictive of the clinical expression and prognosis of the disease, but the molecular mechanisms of the immune responses remain unclear. Recently, we have shown xenobiotic-induced recruitment of nuclear autoantigens to proteasomes in the cell nucleus in cell culture and in animal models in correlation with a unique autoantibody response. In this study, we attempted to validate our findings in patients with SSc. METHODS: Using indirect immunofluorescence microscopy, run-on replication and transcription assays, immunoblotting, and proteasome activity assays, we analyzed the nuclear structure, function, and proteasomal proteolysis in HEp-2 cells treated with xenobiotics or left untreated. Blood dendritic cells (DCs) were isolated from 30 patients with SSc and age- and sex-matched control subjects to determine the subcellular localization of SSc autoantigens in relation to proteasomes. RESULTS: Xenobiotics induced a relocation of the SSc autoantigen DNA topoisomerase I (Scl 70, topo I) to nucleoplasmic clusters, where proteasomes degrade topo I. Colocalization of topo I with proteasomes occurred exclusively in DCs from patients with SSc who developed antibodies against this autoantigen. Neither centromeres nor other SSc autoantigens colocalized with proteasomes in DCs from patients with SSc or from control subjects. CONCLUSION: Alteration of nuclear structure and function by xenobiotics induces recruitment of the nuclear autoantigen topo I for proteasomal processing. This event may, in turn, lead to subsequent presentation of the resulting peptides on the cell surface and the autoimmune responses against topo I in SSc. PMID- 15751093 TI - Cartilage biomarkers in ankylosing spondylitis: relationship to clinical variables and treatment response. AB - OBJECTIVE: Ankylosing spondylitis (AS) is a progressive disease in which chronic inflammation can lead to extensive new bone formation throughout the spine. At present, few measures of the activity or extent of the disease are available. In this study, we sought to determine whether markers of cartilage synthesis and degradation could provide such quantitative measures. METHODS: Serum samples from 23 patients receiving infliximab treatment for AS were obtained at baseline and at weeks 2, 6, 14, and 22. Patients were stratified with respect to joint involvement and baseline levels of inflammatory markers, and responders were defined according to the Assessments in Ankylosing Spondylitis 20% criteria. Serial measurements of interferon-gamma, tumor necrosis factor alpha, transforming growth factor beta (TGFbeta), interleukin-10 (IL-10), and IL-1 were done at each time point. The following biomarkers were measured by enzyme-linked immunosorbent assay: the proteoglycan aggrecan 846 epitope, a marker of cartilage turnover; C-propeptide of type II collagen (CPII), a biosynthesis marker; and the Col2-3/4(long mono) (C2C) and Col2-3/4(short) (C1-2C) neoepitopes, reflecting collagen cleavage of type II collagen and type I/type II collagen, respectively. RESULTS: At baseline, patients with AS demonstrated significant elevations in serum levels of CPII, the 846 epitope, and the CPII-to-C2C (CPII:C2C) ratio (but not C2C or C1-2C) compared with normal controls. Of the biomarkers examined, only CPII:C2C showed a correlation with the C-reactive protein (CRP) level. Among the biomarker-cytokine relationships, TGFbeta demonstrated a trend toward a positive correlation with the 846 epitope. CONCLUSION: In AS, elevated serum levels of CPII and the 846 epitope may be related to biosynthetic turnover of hyaline cartilage and the intervertebral discs but may also reflect progressive bone formation as a result of endochondral ossification. The correlation of the CPII:C2C ratio with CRP suggests that the CPII:C2C ratio might prove to be a useful marker of disease activity in AS. PMID- 15751094 TI - Chondroitins 4 and 6 sulfate in osteoarthritis of the knee: a randomized, controlled trial. AB - OBJECTIVE: To determine whether chondroitin sulfate (CS) is effective in inhibiting cartilage loss in knee osteoarthritis (OA). METHODS: In this randomized, double-blind, placebo-controlled trial, 300 patients with knee OA were recruited from an outpatient clinic, from private practices, and through advertisements. Study patients were randomly assigned to receive either 800 mg CS or placebo once daily for 2 years. The primary outcome was joint space loss over 2 years as assessed by a posteroanterior radiograph of the knee in flexion; secondary outcomes included pain and function. RESULTS: Of 341 patients screened, 300 entered the study and were included in the intent-to-treat analysis. The 150 patients receiving placebo had progressive joint space narrowing, with a mean +/- SD joint space loss of 0.14 +/- 0.61 mm after 2 years (P = 0.001 compared with baseline). In contrast, there was no change in mean joint space width for the 150 patients receiving CS (0.00 +/- 0.53 mm; P not significant compared with baseline). Similar results were found for minimum joint space narrowing. The differences in loss of joint space between the two groups were significant for mean joint space width (0.14 +/- 0.57 mm; P = 0.04) and for minimum joint space width (0.12 +/- 0.52 mm; P = 0.05). CS was well tolerated, with no significant differences in rates of adverse events between the two groups. CONCLUSION: While there was no significant symptomatic effect in this study, long-term treatment with CS may retard radiographic progression in patients with OA of the knee. However, the clinical relevance of the observed structural results has to be further evaluated, and further studies are needed to confirm the structural effects of CS. PMID- 15751095 TI - Therapeutic efficacy of humanized recombinant anti-interleukin-6 receptor antibody in children with systemic-onset juvenile idiopathic arthritis. AB - OBJECTIVE: To investigate the safety and efficacy of a recombinant human anti interleukin-6 (anti-IL-6) receptor monoclonal antibody (MRA) that indirectly inhibits the effects of IL-6 in children with systemic-onset juvenile idiopathic arthritis (JIA) refractory to high-dose, long-term corticosteroids. METHODS: An individual escalating-dose trial was conducted in 11 children with active systemic-onset JIA who met the inclusion criteria. All were first administered an intravenous dose of 2 mg/kg MRA. Each child without active inflammation was given a second identical dose 2 weeks later and a third identical dose 2 weeks after the second dose. Children with disease flares according to laboratory marker values received a 4-mg/kg dose. Those without disease flares at this dose received a second 4-mg/kg dose 2 weeks later and a third 4-mg/kg dose 2 weeks after the second dose, while those with active inflammation received an additional 3 doses of 8 mg/kg MRA. Efficacy was evaluated every 2 weeks according to responses on the JIA core set of improvement criteria and the results of laboratory tests. RESULTS: MRA abruptly reduced disease activity in 10 of the 11 children, as assessed by the occurrence of febrile episodes, active arthritis, scores on the Childhood Health Assessment Questionnaire, and levels of acute phase reactants. However, levels of inflammatory reactants fluctuated until the proper MRA dose for each child was reached. Two weeks after the third fixed dose of MRA, 90.9% of all patients had a 30% improvement response, 90.9% had a 50% improvement response, and 63.6% had a 70% improvement response. CONCLUSION: MRA treatment of children with active systemic disease results in clinical improvement and in normalized levels of acute-phase reactants. MRA was safe and well tolerated and provided greater clinical benefit than conventional corticosteroids, considering the ill effects of IL-6 and adverse events. PMID- 15751096 TI - Women in academic rheumatology. PMID- 15751097 TI - Cardiovascular death in rheumatoid arthritis: a population-based study. AB - OBJECTIVE: To determine whether systemic inflammation confers any additional risk for cardiovascular death among patients with rheumatoid arthritis (RA), after adjusting for traditional cardiovascular risk factors and comorbidities. METHODS: Using the population-based data resources of the Rochester Epidemiology Project, we assembled an incidence cohort of all Rochester, Minnesota residents ages >or=18 years who first fulfilled the American College of Rheumatology 1987 criteria for RA between January 1, 1955 and January 1, 1995. All subjects were followed up longitudinally through their complete (inpatient, outpatient) medical records, beginning at age 18 years and continuing until death, migration, or January 1, 2001. Detailed information on the occurrence of various cardiovascular risk factors (personal history of coronary heart disease [CHD], congestive heart failure, smoking, hypertension, dyslipidemia, body mass index [BMI], diabetes mellitus, menopausal status) as well as indicators of systemic inflammation and RA disease severity (rheumatoid factor [RF] seropositivity, erythrocyte sedimentation rate [ESR], joint swelling, radiographic changes, RA nodules, RA complications, RA treatments, disease duration) and comorbidities were collected on all subjects. Causes of death were ascertained from death certificates and medical records. Cox regression models were used to estimate the independent predictors of cardiovascular death. RESULTS: This inception cohort comprised a total of 603 RA patients whose mean age was 58 years, of whom 73% were women. During a mean followup of 15 years, 354 patients died and cardiovascular disease was the primary cause of death in 176 patients. Personal history of CHD, smoking, hypertension, low BMI, and diabetes mellitus, as well as comorbidities, including peripheral vascular disease, cerebrovascular disease, chronic pulmonary disease, dementia, ulcers, malignancies, renal disease, liver disease, and history of alcoholism, were all significant risk factors for cardiovascular death (P < 0.01 for each). Multivariable Cox regression analyses, controlled for cardiovascular risk factors and comorbidities, revealed that the risk of cardiovascular death was significantly higher among RA patients with at least 3 ESR values of >or=60 mm/hour (hazard ratio [HR] 2.03, 95% confidence interval [95% CI] 1.45-2.83), RA vasculitis (HR 2.41, 95% CI 1.00-5.81), and RA lung disease (HR 2.32, 95% CI 1.11 4.84). CONCLUSION: These results indicate that markers of systemic inflammation confer a statistically significant additional risk for cardiovascular death among patients with RA, even after controlling for traditional cardiovascular risk factors and comorbidities. PMID- 15751098 TI - Monastrol, a selective inhibitor of the mitotic kinesin Eg5, induces a distinctive growth profile of dendrites and axons in primary cortical neuron cultures. AB - Various factors including some motor proteins regulate microtubule (MT) transport and influence the formation of neuronal processes. Eg5, a slow and non-processive (+)-end directed motor molecule, is expressed in developing and differentiated neurons. However, how Eg5 works in neurons is still elusive. Thus, we treated primary rat cortical neuron cultures with monastrol, a specific inhibitor of Eg5, to investigate its role in neurons. Immature neurons treated with monastrol extended longer processes than control within a few hours. After 3 days, immature neurons treated with monastrol had longer dendrites but slightly shorter axons than control. This difference in growth between dendrites and axons became more prominent as the cells differentiated until 5 days. Interestingly, MT distributions in the cell bodies of monastrol-treated neurons appeared somewhat circular surrounding the nucleus, while MTs in the cell bodies of control neurons were primarily distributed in the MT organizing center (MTOC) just beside the nucleus. In mature neurons, monastrol treatment induced the axonal clusters of tubulins, grossly not affecting dendrites. Taken together, we conclude that Eg5 acts distinctively on dendrites and axons in neurons and suggest a putative model of how Eg5 works distinctively on dendrites and axons. PMID- 15751099 TI - Preparing to move: assembly of the MSP amoeboid motility apparatus during spermiogenesis in Ascaris. AB - We exploited the rapid, inducible conversion of non-motile Ascaris spermatids into crawling spermatozoa to examine the pattern of assembly of the MSP motility apparatus that powers sperm locomotion. In live sperm, the first detectable motile activity is the extension of spikes and, later, blebs from the cell surface. However, examination of cells by EM revealed that the formation of surface protrusions is preceded by assembly of MSP filament tails on the membranous organelles in the peripheral cytoplasm. These organelle-associated filament meshworks assemble within 30 sec after induction of spermiogenesis and persist until the membranous organelles are sequestered into the cell body when the lamellipod extends. The filopodia-like spikes, which are packed with bundles of filaments, extend and retract rapidly but last only a few seconds before giving way to, or converting into, blebs. Coalescence of these blebs, each supported by a dense mesh of filaments, often initiates lamellipod extension, which culminates in the formation of the robust, dynamic MSP fiber complexes that generate sperm motility. The same membrane phosphoprotein that orchestrates assembly of the fiber complexes at the leading edge of the lamellipod of mature sperm is also found at all sites of filament assembly during spermiogenesis. The orderly progression of steps that leads to construction of a functional motility apparatus illustrates the precise spatio-temporal control of MSP filament assembly in the developing cell and highlights the remarkable similarity in organization and plasticity shared by the MSP cytoskeleton and the actin filament arrays in conventional crawling cells. PMID- 15751100 TI - Electrochemical hydrogenation of [Ru(bpy)2(napy-kappaN)(CO)]2+: inhibition of reductive Ru--CO bond cleavage by a ruthenacycle. PMID- 15751101 TI - A color sensor for catecholamines. PMID- 15751102 TI - Total synthesis of the marine natural product rac-dibromophakellstatin. PMID- 15751103 TI - NMR spectroscopic detection of protein protons and longitudinal relaxation rates between 0.01 and 50 MHz. PMID- 15751104 TI - Current literature in mass spectrometry. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of mass spectrometry. Each bibliography is divided into 11 sections: 1 Books, Reviews & Symposia; 2 Instrumental Techniques & Methods; 3 Gas Phase Ion Chemistry; 4 Biology/Biochemistry: Amino Acids, Peptides & Proteins; Carbohydrates; Lipids; Nucleic Acids; 5 Pharmacology/Toxicology; 6 Natural Products; 7 Analysis of Organic Compounds; 8 Analysis of Inorganics/Organometallics; 9 Surface Analysis; 10 Environmental Analysis; 11 Elemental Analysis. Within each section, articles are listed in alphabetical order with respect to author (4 Weeks journals - Search completed at 12th. Jan. 2005). PMID- 15751105 TI - Dynamic kinetic resolution during a cascade reaction on substrates with chiral all-carbon quaternary centers. PMID- 15751106 TI - Estimation of the absolute internal-rotation entropy of molecules with two torsional degrees of freedom from stochastic simulations. AB - A method of statistical estimation is applied to the problem of evaluating the absolute entropy of internal rotation in a molecule with two torsional degrees of freedom. The configurational part of the entropy is obtained as that of the joint probability density of an arbitrary form represented by a two-dimensional Fourier series, the coefficients of which are statistically estimated using a sample of the torsional angles of the molecule obtained by a stochastic simulation. The internal rotors in the molecule are assumed to be attached to a common frame, and their reduced moments of inertia are initially calculated as functions of the two torsional angles, but averaged over all the remaining internal degrees of freedom using the stochastic-simulation sample of the atomic configurations of the molecule. The torsional-angle dependence of the reduced moments of inertia can be also averaged out, and the absolute internal-rotation entropy of the molecule is obtained in a good approximation as the sum of the configurational entropy and a kinetic contribution fully determined by the averaged reduced moments of inertia. The method is illustrated using Monte Carlo simulations of isomers of stilbene and halogenated derivatives of propane. The two torsional angles in cis-stilbene are found to be much more strongly correlated than those in trans-stilbene, while the degree of the angular correlation in propane increases strongly on substitution of hydrogen atoms with chlorine. PMID- 15751107 TI - Collision-induced fragmentation of negative ions from N-linked glycans derivatized with 2-aminobenzoic acid. AB - N-Linked glycans from bovine ribonuclease B, chicken ovalbumin, bovine fetuin, porcine thyroglobulin and human alpha(1)-acid glycoprotein were derivatized with 2-aminobenzoic acid by reductive amination and their tandem mass spectra were recorded by negative ion electrospray ionization with a quadrupole time-of-flight mass spectrometer. Derivatives were also prepared from 2-amino-5-methyl- and 2 amino-4,5-dimethoxybenzoic acid in order to confirm the identity of fragment ions containing the reducing terminus. Major fragments from the [M - H](-) ions from the neutral glycans retained the derivative (Y-type cleavages) and provided information on sequence and branching. Other major fragments were products of A type cross-ring cleavages giving information on antenna structure. Singly doubly and triply charged ions were formed from sialylated glycans. They produced major fragments by loss of sialic acid and a series of singly charged ions that were similar to those from the neutral analogues. Doubly charge ions were also produced by the neutral glycans and were fragmented to form product ions with one and two charges. Again, the fragment ions with a single charge were similar to those from the singly charged parents, but branching information was less obvious because of the occurrence of more abundant ions produced by multiple cleavages. Detection limits were around 200 fmol (3 : 1 signal-to-noise ratio). PMID- 15751108 TI - Theoretical study for the reaction of CH3OCl with Cl atom. AB - A direct dynamics method is employed to study the kinetics of the multiple channel reaction CH(3)OCl + Cl. The potential energy surface (PES) information is explored from ab initio calculations. Two reaction channels, Cl- and H abstractions, have been identified. The optimized geometries and frequencies of the stationary points and the minimum-energy paths (MEPs) are calculated at the MP2 level of theory using the 6-311G(d, p) and cc-pVTZ basis sets, respectively. The single-point energies along the MEPs are further refined at the G3(MP2)//MP2/6-311G(d, p), G3//MP2/6-311G(d, p), as well as by the multicoefficient correlation method based on QCISD (MC-QCISD) using the MP2/cc pVTZ geometries. The enthalpies of formation for the species CH(3)OCl and CH(2)OCl are calculated via isodesmic reactions. The rate constants of the two reaction channels are evaluated by using the variational transition-state theory over a wide range of temperature, 220-2200 K. The calculated rate constants exhibit the slightly negative temperature dependence and show good agreement with the available experimental data at room temperature at the G3(MP2)//MP2/6-311G(d, p) level. The present calculations indicate that the two channels are competitive at low temperatures while H-abstraction plays a more important role with the increase of temperature. The calculated k(1a)/k(1) ratio of 0.5 at 298 K is in general agreement with the experimental one, 0.8 +/- 0.2. The high rate constant for CH(3)OCl + Cl shows that removal by reaction with Cl atom is a potentially important loss process for CH(3)OCl in the polar stratosphere. PMID- 15751109 TI - Efficient constrained multiple sequence alignment with performance guarantee. AB - The constrained multiple sequence alignment problem is to align a set of sequences of maximum length n subject to a given constrained sequence, which arises from some knowledge of the structure of the sequences. This paper presents new algorithms for this problem, which are more efficient in terms of time and space (memory) than the previous algorithms, and with a worst-case guarantee on the quality of the alignment. Saving the space requirement by a quadratic factor is particularly significant as the previous O(n4)-space algorithm has limited application due to its huge memory requirement. Experiments on real data sets confirm that our new algorithms show improvements in both alignment quality and resource requirements. PMID- 15751110 TI - Protein folding on the hexagonal lattice in the HP model. AB - In this paper, we introduce the 2D hexagonal lattice as a biologically meaningful alternative to the standard square lattice for the study of protein folding in the HP model. We show that the hexagonal lattice alleviates the "sharp turn" problem and models certain aspects of the protein secondary structure more realistically. We present a 1/6-approximation and a clustering heuristic for protein folding on the hexagonal lattice. In addition to these two algorithms, we also implement a Monte Carlo Metropolis algorithm and a branch-and-bound partial enumeration algorithm, and conduct experiments to compare their effectiveness. PMID- 15751111 TI - Optimizing long intrinsic disorder predictors with protein evolutionary information. AB - Protein existing as an ensemble of structures, called intrinsically disordered, has been shown to be responsible for a wide variety of biological functions and to be common in nature. Here we focus on improving sequence-based predictions of long (>30 amino acid residues) regions lacking specific 3-D structure by means of four new neural-network-based Predictors Of Natural Disordered Regions (PONDRs): VL3, VL3H, VL3P, and VL3E. PONDR VL3 used several features from a previously introduced PONDR VL2, but benefitted from optimized predictor models and a slightly larger (152 vs. 145) set of disordered proteins that were cleaned of mislabeling errors found in the smaller set. PONDR VL3H utilized homologues of the disordered proteins in the training stage, while PONDR VL3P used attributes derived from sequence profiles obtained by PSI-BLAST searches. The measure of accuracy was the average between accuracies on disordered and ordered protein regions. By this measure, the 30-fold cross-validation accuracies of VL3, VL3H, and VL3P were, respectively, 83.6 +/- 1.4%, 85.3 +/- 1.4%, and 85.2 +/- 1.5%. By combining VL3H and VL3P, the resulting PONDR VL3E achieved an accuracy of 86.7 +/ 1.4%. This is a significant improvement over our previous PONDRs VLXT (71.6 +/- 1.3%) and VL2 (80.9 +/- 1.4%). The new disorder predictors with the corresponding datasets are freely accessible through the web server at http://www.ist.temple.edu/disprot. PMID- 15751112 TI - Bayesian network learning with feature abstraction for gene-drug dependency analysis. AB - Combined analysis of the microarray and drug-activity datasets has the potential of revealing valuable knowledge about various relations among gene expressions and drug activities in the malignant cell. In this paper, we apply Bayesian networks, a tool for compact representation of the joint probability distribution, to such analysis. For the alleviation of data dimensionality problem, the huge datasets were condensed using a feature abstraction technique. The proposed analysis method was applied to the NCI60 dataset (http://discover.nci.nih.gov) consisting of gene expression profiles and drug activity patterns on human cancer cell lines. The Bayesian networks, learned from the condensed dataset, identified most of the salient pairwise correlations and some known relationships among several features in the original dataset, confirming the effectiveness of the proposed feature abstraction method. Also, a survey of the recent literature confirms the several relationships appearing in the learned Bayesian network to be biologically meaningful. PMID- 15751113 TI - Adaptive control of hybridization noise in DNA sequencing-by-hybridization. AB - We consider the problem of sequence reconstruction in sequencing-by-hybridization in the presence of spectrum errors. As suggested by intuition, and reported in the literature, false-negatives (i.e., missing spectrum probes) are by far the leading cause of reconstruction failures. In a recent paper we have described an algorithm, called "threshold-theta", designed to recover from false negatives. This algorithm is based on overcompensating for missing extensions by allowing larger reconstruction subtrees. We demonstrated, both analytically and with simulations, the increasing effectiveness of the approach as the parameter theta grows, but also pointed out that for larger error rates the size of the extension trees translates into an unacceptable computational burden. To obviate this shortcoming, in this paper we propose an adaptive approach which is both effective and efficient. Effective, because for a fixed value of theta it performs as well as its single-threshold counterpart, efficient because it exhibits substantial speed-ups over it. The idea is that, for moderate error rates a small fraction of the target sequence can be involved in error recovery; thus, expectedly the remainder of the sequence is reconstructible by the standard noiseless algorithm, with the provision to switch to operation with increasingly higher thresholds after detecting failure. This policy generates interesting and complex interplays between fooling probes and false negatives. These phenomena are carefully analyzed for random sequences and the results are found to be in excellent agreement with the simulations. In addition, the experimental algorithmic speed-ups of the multithreshold approach are explained in terms of the interaction amongst the different threshold regimes. PMID- 15751115 TI - Protein side-chain packing problem: a maximum edge-weight clique algorithmic approach. AB - "Protein Side-chain Packing" has an ever-increasing application in the field of bio-informatics, dating from the early methods of homology modeling to protein design and to the protein docking. However, this problem is computationally known to be NP-hard. In this regard, we have developed a novel approach to solve this problem using the notion of a maximum edge-weight clique. Our approach is based on efficient reduction of protein side-chain packing problem to a graph and then solving the reduced graph to find the maximum clique by applying an efficient clique finding algorithm developed by our co-authors. Since our approach is based on deterministic algorithms in contrast to the various existing algorithms based on heuristic approaches, our algorithm guarantees of finding an optimal solution. We have tested this approach to predict the side-chain conformations of a set of proteins and have compared the results with other existing methods. We have found that our results are favorably comparable or better than the results produced by the existing methods. As our test set contains a protein of 494 residues, we have obtained considerable improvement in terms of size of the proteins and in terms of the efficiency and the accuracy of prediction. PMID- 15751117 TI - A tabu search algorithm for post-processing multiple sequence alignment. AB - Tabu search is a meta-heuristic approach that is proven to be useful in solving combinatorial optimization problems. We implement the adaptive memory features of tabu search to refine a multiple sequence alignment. Adaptive memory helps the search process to avoid local optima and explores the solution space economically and effectively without getting trapped into cycles. The algorithm is further enhanced by introducing extended tabu search features such as intensification and diversification. The neighborhoods of a solution are generated stochastically and a consistency-based objective function is employed to measure its quality. The algorithm is tested with the datasets from BAliBASE benchmarking database. We have observed through experiments that tabu search is able to improve the quality of multiple alignments generated by other software such as ClustalW and T-Coffee. The source code of our algorithm is available at http://www.bii.a star.edu.sg/~tariq/tabu/. PMID- 15751116 TI - Active site prediction for comparative model structures with thematics. AB - THEMATICS (Theoretical Microscopic Titration Curves) is a simple, reliable computational predictor of the active sites of enzymes from structure. Our method, based on well-established Finite Difference Poisson-Boltzmann techniques, identifies the ionisable residues with anomalous predicted titration behavior. A cluster of two or more such perturbed residues is a very reliable predictor of the active site. The protein does not have to bear any resemblance in sequence or structure to any previously characterized protein, but the method does require the three-dimensional structure. We now present evidence that THEMATICS can also locate the active site in structures built by comparative modeling from similar structures. Results are given for a total of 21 sets of proteins, including 21 templates and 83 comparative model structures. Detailed results are presented for three sets of orthologous proteins (Triosephosphate isomerase, 6-Hydroxymethyl 7,8-dihydropterin pyrophosphokinase, and Aspartate aminotransferase) and for one set of human homologues of Aldose reductase with different functions. THEMATICS correctly locates the active site in the model structures. This suggests that the method can be applicable to a much larger set of proteins for which an experimentally determined structure is unavailable. With a few exceptions, the predicted active sites in the comparative model structures are similar to that of the corresponding template structure. PMID- 15751118 TI - Evolution of relative synonymous codon usage in Human Immunodeficiency Virus type 1. AB - Mutation in Human Immunodeficiency Virus type-1 (HIV-1) is extremely rapid, a consequence of a low-fidelity viral reverse transcription process. The envelope gene has been shown to accumulate substitutions at a rate of approximately 1% per year and can frequently spend a long time in the host (approximately 10 years). The relative synonymous codon usage (RSCU) in HIV-1 is known to be different from that of the human host. However, by reengineering the protein coding sequences of HIV-1 to reflect the RSCU patterns observed in humans, a large increase in protein expression is observed. It is reasonable to suggest that within a host there may be a selective drive for change in the RSCU of HIV-1 towards human RSCU. To test this hypothesis we analyzed HIV-1 partial envelope sequences from eight patients sampled serially in time. For each sequence, an RSCU table was constructed. Sequences were labelled as "early" or "late" depending on whether they were sampled before or after the mid-point of the study. Using the RSCU values as descriptor variables, a Principal Components Analysis (PCA) was performed. The first three components clearly discriminated between early and late sequences. We also constructed pooled groupwise RSCU tables for early and late sequences. The viral RSCU values of each of the groups were correlated with human RSCU. If there is selection for host-adaptation in RSCU, we expect that "late" viral RSCUs would tend to be more highly correlated with human RSCU than "early" viral RSCUs. In fact, tests of significance suggest that this is the case. However, closer examination of the data revealed that the apparent trend towards human RSCU can be attributed to the homogenization of the codon usage by mutation pressure rather than host adaptation. PMID- 15751119 TI - Integration of bioinformatics and computational biology to understand protein-DNA recognition mechanism. AB - Transcription factors play essential role in the gene regulation in higher organisms, binding to multiple target sequences and regulating multiple genes in a complex manner. In order to decipher the mechanism of gene regulation, it is important to understand the molecular mechanism of protein-DNA recognition. Here we describe a strategy to approach this problem, using various methods in bioinformatics and computational biology. We have used a knowledge-based approach, utilizing rapidly increasing structural data of protein-DNA complexes, to derive empirical potential functions for the specific interactions between bases and amino acids as well as for DNA conformation, from the statistical analyses on the structural data. Then these statistical potentials are used to quantify the specificity of protein-DNA recognition. The quantification of specificity has enabled us to establish the structure-function analysis of transcription factors, such as the effects of binding cooperativity on target recognition. The method is also applied to real genome sequences, predicting potential target sites. We are also using computer simulations of protein-DNA interactions and DNA conformation in order to complement the empirical method. The integration of these approaches together will provide deeper insight into the mechanism of protein-DNA recognition and improve the target prediction of transcription factors. PMID- 15751120 TI - Proteomic analysis of the wing imaginal discs of Drosophila melanogaster. AB - We have combined high-resolution two-dimensional (2-D) gel electrophoresis and mass spectrometry with the aim of identifying proteins represented in the 2-D gel database of the wing imaginal discs of Drosophila melanogaster. First, we obtained a high-resolution 2-D gel pattern of [35S]methionine + [35S]cysteine labeled polypeptides of Schneider cells, a permanent cell line of Drosophila embryonic origin, and compared it with the standard pattern of polypeptides of the wing imaginal disc. These studies reveal qualitative and quantitative differences between the two samples, but have more than 600 polypeptides in common. Second, we carried out preparative 2-D polyacrylamide gel electrophoresis using Schneider cells mixed with radioactively labeled wing imaginal discs in order to isolate some of the shared polypeptides and characterize them by matrix assisted laser desorption/ionization-time of flight MALDI-TOF analysis. Using this strategy we identified 100 shared proteins represented in the database, and in each case confirmed their identity by MALDI-TOF/TOF analysis. PMID- 15751121 TI - Effect of dietary calcium: Phosphorus ratio on bone mineralization and intestinal calcium absorption in ovariectomized rats. AB - We investigated the effect of dietary calcium:phosphorus (Ca:P) ratio on bone mineralization and intestinal Ca absorption in ovariectomized (OVX) rat models of osteoporosis and sham-operated rats. Thirty 12-wk-old female Wistar rats were divided into three groups of OVX rats and three groups of sham rats. Thirty days after the adaptation period, OVX rats and sham rats were fed a diet formulated Ca:P, 1:0.5, 1:1 or 1:2 (each diet containing 0.5% Ca), respectively for 42 d. In both sham and OVX rats, serum osteocalcin, a marker of bone turnover, was increased by decreasing Ca:P ratio (1:2). In contrast, rats fed the Ca:P = 1:0.5 diet (dietary P restriction) suppressed the increased serum parathyroid hormone, osteocalcin and urinary deoxypyridinoline, and increased Ca absorption in both sham and OVX rats compared to the Ca:P = 1:1 and 1:2 diets. Especially, in OVX rats, the decreased bone mineral density of the fifth lumbar was also suppressed when rats were fed the Ca:P = 1:0.5 diet. These results indicated that the elevation of dietary Ca:P ratio may inhibit bone loss and increase intestinal Ca absorption in OVX rats. PMID- 15751122 TI - Citrus nobiletin inhibits azoxymethane-induced large bowel carcinogenesis in rats. AB - The inhibitory effects of dietary feeding of citrus nobiletin on azoxymethane (AOM)-induced rat colon carcinogenesis using a long-term bioassay were investigated. Five-week old male F344 rats were initiated with two weekly subcutaneous injections of AOM (20 mg/kg bw) to induce colonic tumors. They were also given the diets containing 0.01% or 0.05% nobiletin for 34 weeks, starting one week after the last dosing of AOM. At the end of the study, the incidence of colonic adenocarcinoma were 67% in the AOM alone group, 55% in the AOM-->0.01% nobiletin group, 35% (p<0.05) in the AOM-->0.05% nobiletin group. Also, nobiletin feeding reduced the cell-proliferation activity, increased the apoptotic index, and decreased the prostaglandin E2 content in colonic adenocarcinoma and/or colonic mucosa. These findings might suggest that citrus nobiletin has chemopreventive ability against AOM-induced rat colon carcinogenesis. PMID- 15751123 TI - Skeletal Muscle wasting and contractile performance in septic rats. AB - We investigated the temporal effects of sepsis on muscle wasting and function in order to study the contribution of wasting to the decline in muscle function; we also studied the fiber-type specificity of this muscle wasting. Sepsis was induced by injecting rats intraperitoneally with a zymosan suspension. At 2 h and at 2, 6, and 11 days after injection, muscle function was measured using in situ electrical stimulation, Zymosan injection induced severe muscle wasting compared to pair-fed and ad libitum fed controls. At 6 days, isometric force-generating capacity was drastically reduced in zymosan-treated rats. We conclude that this was fully accounted fo by the reduction of muscle mas. At day 6, we also observed increased activity of the 20S proteasome in gastrocnemius but not soleus muscle from septic rats. In tibialis anterior but not in soleus, muscle wasting occurred in a fiber-type specific fashion, i.e., the reduction in cross-sectional area was significantly smaller in type 1 than type 2A and 2B/X fibers. These findings suggest that both the inherent function of a muscle and the muscle fiber-type distribution affect the responsiveness to catabolic signals. PMID- 15751124 TI - What kind of farming works best? PMID- 15751125 TI - India's R&D: reaching for the top. PMID- 15751127 TI - Umbilical-cord blood for transplantation in adults. PMID- 15751128 TI - ACE inhibition in stable coronary artery disease. PMID- 15751129 TI - ACE inhibition in stable coronary artery disease. PMID- 15751130 TI - Premature birth and insulin resistance. PMID- 15751131 TI - Radial-artery coronary bypass grafts. PMID- 15751132 TI - Pharmacists and emergency contraception. PMID- 15751133 TI - Pharmacists and emergency contraception. PMID- 15751134 TI - A novel transcriptional autoregulatory loop enhances expression of the Pantoea stewartii subsp. stewartii Hrp type III secretion system. AB - The hrp type III secretion regulon of Pantoea stewartii is regulated by a cascade involving the HrpX/HrpY two-component system, the HrpS enhancer-binding protein and the HrpL alternate sigma factor. hrpXY is both constitutive and autoregulated; HrpY controls hrpS; and HrpS activates hrpL. These regulatory genes are arranged in the order hrpL, hrpXY and hrpS and constitute three operons. This study describes a novel autoregulatory loop involving HrpS. Genetic experiments using a chromosomal hrpS-lacZ fusion demonstrated that ectopic expression of HrpS increases hrpS transcription and that this effect is blocked by polar mutations in hrpXY and hrpL and by a nonpolar mutation in hrpY. RT-PCR and Northern blot analysis revealed a hrpL-hrpXY polycistronic mRNA. These results suggest that HrpS-mediated autoregulation is due to activation of hrpS by increased levels of HrpY resulting from read-through transcription of hrpXY from the hrpL promoter. This novel autoregulatory loop may serve to rapidly induce hrp genes during infection and to compensate for negative regulatory mechanisms that keep the regulon off in the insect vector. PMID- 15751135 TI - Single dose theory. PMID- 15751137 TI - Time to move on. More questions about single-dose nevirapine. PMID- 15751136 TI - Single-dose jeopardizes long-term therapy. PMID- 15751139 TI - Pediatric AIDS Foundation on single-dose nevirapine. PMID- 15751138 TI - Impact of the second Bush Administration on national HIV/AIDS public policy. PMID- 15751140 TI - Comments by Global Strategies for HIV Prevention on nevirapine resistance. PMID- 15751141 TI - The Rolls-Royce and the Tuk-Tuk. PMID- 15751142 TI - Chemical and morphological surface modification of PLA50: optimization for human skin cellcultures under biodegradable polymers. PMID- 15751143 TI - Light-emitting diodes as an illumination source for plants: a review of research at Kennedy Space Center. AB - The provision of sufficient light is a fundamental requirement to support long term plant growth in space. Several types of electric lamps have been tested to provide radiant energy for plants in this regard, including fluorescent, high pressure sodium, and metal halide lamps. These lamps vary in terms of spectral quality, which can result in differences in plant growth and morphology. Current lighting research for space-based plant culture is focused on innovative lighting technologies that demonstrate high electrical efficiency and reduced mass and volume. Among the lighting technologies considered for space are light-emitting diodes (LEDs). The combination of red and blue LEDs has proven to be an effective lighting source for several crops, yet the appearance of plants under red and blue lighting is purplish gray, making visual assessment of plant health difficult. Additional green light would make the plant leaves appear green and normal, similar to a natural setting under white light, and may also offer psychological benefits for the crew. The addition of 24% green light (500-600 nm) to red and blue LEDs enhanced the growth of lettuce plants compared with plants grown under cool white fluorescent lamps. Coincidentally, these plants grown under additional green light would have the additional aesthetic appeal of a green appearance. PMID- 15751144 TI - Porous media matric potential and water content measurements during parabolic flight. AB - Control of water and air in the root zone of plants remains a challenge in the microgravity environment of space. Due to limited flight opportunities, research aimed at resolving microgravity porous media fluid dynamics must often be conducted on Earth. The NASA KC-135 reduced gravity flight program offers an opportunity for Earth-based researchers to study physical processes in a variable gravity environment. The objectives of this study were to obtain measurements of water content and matric potential during the parabolic profile flown by the KC 135 aircraft. The flight profile provided 20-25 s of microgravity at the top of the parabola, while pulling 1.8 g at the bottom. The soil moisture sensors (Temperature and Moisture Acquisition System: Orbital Technologies, Madison, WI) used a heat-pulse method to indirectly estimate water content from heat dissipation. Tensiometers were constructed using a stainless steel porous cup with a pressure transducer and were used to measure the matric potential of the medium. The two types of sensors were placed at different depths in a substrate compartment filled with 1-2 mm Turface (calcined clay). The ability of the heat pulse sensors to monitor overall changes in water content in the substrate compartment decreased with water content. Differences in measured water content data recorded at 0, 1, and 1.8 g were not significant. Tensiometer readings tracked pressure differences due to the hydrostatic force changes with variable gravity. The readings may have been affected by changes in cabin air pressure that occurred during each parabola. Tensiometer porous membrane conductivity (function of pore size) and fluid volume both influence response time. Porous media sample height and water content influence time-to-equilibrium, where shorter samples and higher water content achieve faster equilibrium. Further testing is needed to develop these sensors for space flight applications. PMID- 15751145 TI - Nursing shortage, TPAPN funding to top TNA's legislative priorities for 2005. PMID- 15751146 TI - Oral health workforce planning for developed countries. PMID- 15751147 TI - Antioxidative polyphenols from berries of Pimenta dioica. AB - The ethyl acetate-soluble part of allspice, berries of Pimenta dicica, showed strong antioxidant activity and radical-scavenging activity against 1,1diphenyl-2 picrylhydrazl (DPPH) radical. From the ethyl acetate-soluble part, two new compounds, 5-galloyloxy-3-4-dihydroxypentanoic acid and 5-(5-carboxmethyl-2 oxocyclopenty)3Z-penteny 6-O-galloy-beta-D-glucoside were isolated together with 11 known polyphenols by repeated column chromatography. Their structures were elucidated on the basis of MS and various NMR spectroscopic data. All isolated compounds were evaluated for antioxidative effects on oxidation of methyl linoleate under aeration and heating, anf on peroxidation of liposome induced by 2-2'-azobis-(2-amidinopropane)dihydrocloride (AAPH) as water-soluble initiator along with their radical-scavenging activity against DPPH. Quercetin and its glycoside showed remarkable activity for scavenging DPPH radical and inhibiting peroxidation of liposome. Two new compounds also exhibited strong DPPH radical scavenging activity and inhibitory effect on the peroxidation od liposome as myricetin. PMID- 15751148 TI - Heinrich v. Sweet. PMID- 15751149 TI - Craft v. Vanderbilt University. PMID- 15751150 TI - Morphofunctional basis for recovery of locomotor movements in rats with completely crossed spinal cord. AB - Treadmill training of spinalized rats creates conditions for early appearance of rhythmic locomotor movements of the hind limbs. Recovery of the movements was paralleled by an appropriate structural organization of neurons in the anterior horns of the distal compartment of the spinal cord. PMID- 15751151 TI - Bibeau v. Pacific Northwest Research Foundation. PMID- 15751152 TI - Pegvisomant: new preparation. A last resort in acromegaly. AB - (1) The first-line treatment for acromegaly is transsphenoidal surgery. As an adjunct to surgery, and for patients with inoperable tumours, the first-line drug therapy is a somatostatin analogue (octreotide or lanreotide). (2) Pegvisomant, a growth hormone (GH) receptor antagonist, is licensed for patients who have a poor response to surgery and/or radiation therapy and in whom somatostatin analogue therapy has failed. (3) There are no published comparative trials evaluating pegvisomant as alternative for patients who have already tried a somatostatin analogue. Most of the evaluation data comes from a single randomised, placebo controlled trial in 112 patients treated for three months. It showed that pegvisomant, at a dose of 10 mg to 20 mg/day subcutaneously, normalised the level of insulin-like growth factor type 1 (IGF-1) and improved some symptoms. However, the patients were very heterogeneous, and no information was provided on the efficacy of previous treatments. This trial does not provide enough evidence to support the approved indication. (4) Pegvisomant lowers the IGF-1 level but also increases the GH level. An increase in the size of the pituitary adenoma, due to the resulting hyperfunctioning, was observed in four of the 160 patients treated in clinical trials. (5) Evaluation data on pegvisomant does not resolve the question of possible hepatic toxicity. (6) In practice, pegvisomant therapy is justified for patients with serious complications of acromegaly and who have no other treatment options. But the patients concerned, who are few in number, must be closely monitored and included in an appropriate register. PMID- 15751153 TI - Free radical scavenger protects heart from adriamycin toxicity. PMID- 15751154 TI - Oral BMS-354825 rescues Gleevec-resistant CML. PMID- 15751155 TI - New clues to target duo of chemokines in breast cancer stromal microenvironment. PMID- 15751156 TI - Cancer drug development and translational research are high priorities of special workshop. PMID- 15751157 TI - Gleevec target Abl important for normal immune activation of T cells. PMID- 15751158 TI - [Antitumor effect and apoptosis induction in human cancer cell lines by BRM-SJS]. AB - BACKGROUND & OBJECTIVE: It had been observed that BRM-SJS had antitumor effect in our clinical practice. This study was designed to investigate the antitumor activity of BRM-SJS, and mechanism of its action. METHODS: In vitro antitumor experiments with MTT method, meanwhile cell morphology, flow cytometry, and agarose gel electrophoresis were performed for determining apoptosis in several tumor cell lines. RESULTS: BRM-SJS had antitumor effects on human Suzhou human glioma (SHG-44), breast carcinoma (MCF-7), and human pancreas carcinoma (PANC1) in vitro, the IC50 values of BRM-SJS were 0. 299 mg/ml, 1.853 mg/ml and 9.416 mg/ml respectively. At the 2. 5 mg of BRM-SJS on SHG-44 and MCF-7, marked morphological changes, including cell shrinkage and condensation of chromosomes, were observed with electric microscope. The increase of apoptosis in SHG-44 and MCF-7 cells treated with BRM-SJS extracts 0.625 -2.5 mg for 14 -48 h was observed by Annexin-V/PI flow cytometry analysis. Agarose gel electrophoresis of DNA from SHG-44 and MCF-7 cells treated with BRM-SJS extracts 1.25 -5 mg for 24 h or 48 h showed marked DNA Ladder pattern. CONCLUSION: Antitumor activity of BRM-SJS may be related with inducement of apoptosis of tumor cells. PMID- 15751159 TI - Inhibitor of apoptosis (IAP) rescues migration defect in Rac-deficient flies. PMID- 15751160 TI - Francis Crick, 1916-2004. PMID- 15751161 TI - Digitized serum mass spectrometry peaks diagnostic of prostate cancer. PMID- 15751163 TI - Clinical targeting of VEGF and EGFR effective in non-small cell lung cancer. PMID- 15751162 TI - HIF-1 blockade inhibits gastric cancer growth in mice. PMID- 15751164 TI - Knudson wins 2004 Kyoto Prize. PMID- 15751165 TI - Kelly, Langer. Sherr and Stillman win 2004 GM Awards. PMID- 15751166 TI - Folkman, Hunter, Massague, Vogelstein and Weinberg win 2004 Prince of Asturias Awards. PMID- 15751167 TI - Dextrocardia and atrial fibrillation. PMID- 15751168 TI - Dalteparin: new indication. Prophylaxis in medical patients: no advance. AB - (1) Heparin prophylaxis for medical inpatients who are confined to bed is controversial, because there are no reliable comparative data. Prophylaxis only seems justified for some patients at high risk of pulmonary embolism who have no risk factors for bleeding. (2) The licence of dalteparin, a low-molecular-weight heparin (LMWH), has been extended in France to cover prophylaxis of deep venous thrombosis in patients confined to bed for heart failure, acute respiratory failure, acute infections or acute rheumatological conditions who have at least one other risk factor for venous thromboembolism. (3) Evaluation data in this setting include a placebo-controlled trial but no trials versus unfractionated heparin or enoxaparin (another LMWH already available for this use). (4) The PREVENT trial included 3681 patients matching the characteristics described in the licence. They were randomised to receive (double-blind) daily subcutaneous injections of dalteparin (5000 IU) or placebo for 14 days. There was no difference between the groups in the following outcomes: death, pulmonary embolism and venous thrombosis (incidence below 1% in the placebo group). The results based on an endpoint combining clinical outcomes and phlebographic abnormalities favoured dalteparin. (5) Few data are available on the adverse events occurring in this trial. In other clinical situations, dalteparin has the same adverse effects as other LMWH (thrombocytopenia, hyperkalemia, etc.). (6) In practice, for medical inpatients who are confined to bed, where the thromboembolic risk is low, dalteparin offers no tangible advantages over unfractionated heparin or enoxaparin. The optimal dose for preventing symptomatic thromboembolism and minimising the bleeding risk is unknown. PMID- 15751169 TI - In re Cincinnati Radiation Litigation. PMID- 15751171 TI - Sertraline and venlafaxine: new indication. Prevention of recurrent depression: no advance. AB - (1) An antidepressant that worked in a depressive episode can be used to prevent recurrences in patients who do not have bipolar disorder. Several antidepressants are at least partially effective in this setting. (2) Sertraline and venlafaxine are now authorised in France for the prevention of recurrent depression. (3) Sertraline and short-acting venlafaxine were both more effective than placebo in single trials (although the data on venlafaxine were somewhat weaker than the data on sertraline). Neither trial has been published in detail. There are no trials comparing sertraline or venlafaxine with other antidepressants in this setting. (4) These trials confirmed the known adverse effect profiles of sertraline and venlafaxine. (5) In practice, these licence extensions have no clinical implications for the management of depression. PMID- 15751170 TI - Docetaxel: new indication. First-line treatment of non small-cell lung cancer: no advance. AB - (1) The reference first-line drug therapy for patients with non-operable non small-cell lung cancer is a combination of two cytotoxic agents, one of which is a platinum compound. The survival benefit is no more than a few months. (2) The docetaxel + cisplatin combination has now been authorised in France for first line treatment of locally advanced and metastatic non small-cell lung cancer. Evaluation data includes the results of three comparative trials. (3) In one trial the docetaxel + cisplatin combination was no more effective than the docetaxel + carboplatin combination or the vinorelbine + cisplatin combination on either the survival time (9.4 to 11.3 months) or on other endpoints. (4) Similar results were obtained in a trial versus paclitaxel + cisplatin and gemcitabine + cisplatin (median survival time 8 months in each group). (5) In a trial versus vindesine + cisplatin, the median survival time was longer with docetaxel + cisplatin (11.3 versus 9.6 months). (6) It is difficult to analyse adverse effects in these unblinded trials. Globally, the docetaxel + cisplatin combination did not appear to be safer than the comparator combinations, particularly with regard to serious events. (7) Docetaxel, like paclitaxel, is infused intravenously every three weeks. The comparator combinations tested in the three clinical trials are infused once a week. (8) In practice, for first line treatment of inoperable non small-cell lung cancer, the docetaxel + cisplatin combination is simply one of several options, and offers no advantages in terms of survival or adverse effects. PMID- 15751172 TI - Topical steroid allergy and dependence. AB - (1) When a topical corticosteroid fails to control a skin condition, the first explanations are usually a wrong diagnosis or inadequate drug potency; however, allergy or dependence should also be considered. (2) Contact dermatitis due to a topical corticosteroid is difficult to diagnose as the symptoms are often mixed with those of the underlying skin disease. Allergy to topical corticosteroids can mimic acute eczema or localised acute swelling. The most commonly affected areas are the legs, hands and face. (3) Risk factors include long term, frequent application of topical steroids by patients with leg ulcers, stasis dermatitis, atopic dermatitis or contact dermatitis (especially on the hands). The diagnostic performance of skin tests is controversial. (4) Several studies and other lines of evidence point to rare allergic cross-reactions to topical corticosteroids, undermining the usefulness of switching to a second topical corticosteroid. (5) Sometimes, especially when skin tests are negative, the problem seems to be dependence to the topical corticosteroid rather than allergy. (6) In practice, stopping the steroid treatment completely is sometimes the best solution, although this may prove difficult. PMID- 15751173 TI - Grafting of poly(ethylene oxide)/poly(butylene terephthalate) block copolymers onto hydroxyapatite particles. PMID- 15751174 TI - PEG-PLA hydrogels by stereocomplexation for tissue engineering of cartilage. PMID- 15751175 TI - Preparation of hydrogels by photo-crosslinking of fumaric acid monoethyl ester (fame) functionalized oligomers. PMID- 15751176 TI - Collagen scaffolds for hematopoietic progenitor cell expansion and controlled differentiation. PMID- 15751177 TI - Coxib toxicity: a likely class effect. AB - All coxibs appear to increase the risk of cardiovascular events. In Canada, widespread prescription of rofecoxib and celecoxib was followed by an increase in hospital admissions for gastrointestinal haemorrhage. PMID- 15751178 TI - Accidental child poisoning with methylergometrine intended for the mother. AB - (1) Erroneous administration to an infant of methylergometrine intended to treat postpartum haemorrhage in the mother can have severe repercussions for the child. (2) The risk is particularly high when the mother and newborn are being treated simultaneously immediately after delivery or in the postpartum period. PMID- 15751179 TI - Suicide and antidepressants in children (continued). PMID- 15751180 TI - Adult hypertension: reducing cardiovascular morbidity and mortality. AB - (1) Since our last review of treatments for arterial hypertension in 1999 (Prescrire International no.41), many new data have been published and new antihypertensive drugs have appeared on the market. (2) The working definition of hypertension is unchanged, namely blood pressure of at least 160/95 mm Hg in the general population, and at least 140/80 mm Hg in patients with diabetes and a history of stroke; these figures must be found on several occasions using a standardised method, with the patient at rest. (3) The goals of antihypertensive therapy are to reduce mortality and cardiovascular events, and not simply to drive blood pressure below a fixed (and often controversial) threshold. (4) Some drug and non drug interventions have a positive risk-benefit balance in the long term. (5) When antihypertensive drug therapy is needed, trials based on clinical endpoints show that it is best to start treatment with a single drug. (6) New data support the use of certain thiazide diuretics (chlortalidone, or hydrochlorothiazide if chlortalidone is not available) as first line treatment for most hypertensive patients, including non diabetic adults, diabetic adults, elderly subjects (over 65 years), and stroke patients. Some betablockers and angiotensin-converting-enzyme inhibitors (ACE inhibitor) are second-line alternatives. (7) Assessment of other antihypertensive drugs has also progressed since 1999, including indapamide (thiazide-like diuretic), amlodipine, diltiazem and verapamil (calcium channel blockers), lisinopril (ACE inhibitor), and losartan and valsartan (angiotensin II antagonists). However, these drugs are not as thoroughly evaluated as thiazide diuretics, betablockers and some ACE inhibitors. PMID- 15751181 TI - Stem cell research and its prospects for liver disease treatment. PMID- 15751182 TI - A young man from Peru with fever and abdominal pain. PMID- 15751183 TI - Controlled delivery of methotrexate from channel-protein containing liposomes. PMID- 15751184 TI - Skin branding. PMID- 15751185 TI - A patient's opinion is often valuable. PMID- 15751187 TI - Human African trypanosomiasis. PMID- 15751186 TI - Sequencing the environment. PMID- 15751189 TI - Autosomal recessive polycystic kidney disease. PMID- 15751188 TI - The lonely eye. PMID- 15751190 TI - Founder's syndrome--a serious corporate malady. PMID- 15751191 TI - Chloroquine-induced retinal toxicity. PMID- 15751192 TI - A tale of two industries. PMID- 15751193 TI - Pruritus ani. PMID- 15751194 TI - AIDS fuels ownerless feral dog populations. PMID- 15751195 TI - Snatching victory from the jaws of defeat. PMID- 15751196 TI - Generics prices in SA compare well with those in other countries. PMID- 15751197 TI - Nicotinic acid and HDL-C reduction. PMID- 15751198 TI - GlaxoSmithKline receives European approval for Kivexa. PMID- 15751199 TI - FDA issues public health advisory recommending limited use of COX-2 inhibitors. PMID- 15751200 TI - Barber shaving and blood-borne disease transmission in developing countries. PMID- 15751201 TI - Human rights abuses in Zimbabwe. PMID- 15751202 TI - Pethidine--does familiarity or evidence perpetuate its use? PMID- 15751203 TI - Recommendations pertaining to the use of viral vaccines: influenza, 2005. PMID- 15751204 TI - Research protocols--lessons from ethical review. PMID- 15751205 TI - How equitable is the scaling up of HIV service provision in South Africa? AB - OBJECTIVES: To assess the extent of inequalities in availability and utilisation of HIV services across South Africa. DESIGN: Cross-sectional descriptive study. SETTING: Three districts reflecting different socio-economic conditions, but with similar levels of HIV infection, were purposively sampled. OUTCOME MEASURES: Availability and utilisation of HIV services and management and support structures for programmes were assessed through the collection of secondary data supplemented by site visits. RESULTS: There were marked inequalities in service delivery between the three sites. Compared with two poorer sites, clinics at the urban site had greater availability of HIV services, including voluntary counselling and testing (100% v. 52% and 24% respectively), better uptake of this service (59 v. 9 and 5.5 clients per 1000 adults respectively) and greater distribution of condoms (15.6 v. 8.2 condoms per adult male per year). Extra counsellors had also been employed at the urban site in contrast to the other 2 sites. The urban site also had far more intensive management support and monitoring, with 1 manager per 12 health facilities compared with 1 manager per more than 90 health facilities at the other 2 sites. CONCLUSION: The process of scaling up of HIV services seems to be accentuating inequalities. The urban site in this study was better able to utilise the extra resources. In contrast, the poorer sites have thus far been unable to scale up the response to HIV even with the availability of extra resources. Unless policy makers pay more attention to equity, efficacious interventions may prove to be of limited effectiveness. PMID- 15751206 TI - Market and product assessment of probiotic/prebiotic-containing functional foods and supplements manufactured in South Africa. AB - OBJECTIVES: Probiotic and prebiotic products manufactured in South Africa were identified and health and content claims stated on the labels were evaluated according to available scientific evidence, the proposed South African regulations in the Foodstuffs, Cosmetics and Disinfectants Act (Act No. 54 of 1972, www.doh.gov.za), and microbial assessment. RESULTS: The range of products identified included probiotic- and/or prebiotic-containing supplements (capsules), food items fortified with probiotics and/or prebiotics, and fermented food containing probiotics, e.g. dairy products. Most of the health-related claims on the labels of the identified products do not comply with proposed South African regulations. However, results also indicate that the proposed South African regulations should be reconsidered to include an additional 5 claims, for which scientifically sound evidence is available. The claims regarding probiotic strains, viable cell numbers, prebiotic type and concentration stated on the labels of the products are mostly in line with the proposed South African regulations. The actual viable cell content of 3 out of 5 probiotic supplements readily available on the South African market did not comply with the content claim stated on the label. However, this problem did not seem to affect the inhibitory activity of the probiotic strains against indicator strains isolated from faeces of patients diagnosed with AIDS. To validate this finding in vivo assessments should be implemented before considering the need to include a wider range of prescribed viable cell numbers in the proposed South African regulations. CONCLUSIONS: The proposed South African regulations regarding probiotic- and prebiotic-containing products should be revised based on the results of this research, and the manufacturers of these products should be held responsible for providing the consumer with scientifically sound and legally correct information. PMID- 15751207 TI - Obsessive compulsive disorder--prevalence in Xhosa-speaking schizophrenia patients. AB - Obsessive compulsive disorder (OCD) has been reported in up to 31% of schizophrenia sufferers. This study evaluated the presence of OCD in a Xhosa speaking schizophrenia group. Xhosa patients (N = 509, including 100 sibships) with schizophrenia were recruited from hospital and community settings. The patients underwent a structured clinical interview for the presence of lifetime co-morbid schizophrenia and OCD. Only 3 patients (0.5%) fulfilled criteria for OCD. No concordance for OCD was noted in the sibship group. Our findings differ from those in other parts of the world, and if replicated, might suggest unique protective environmental or genetic factors for OCD in certain ethnic groups. PMID- 15751208 TI - Non-closure of peritoneal surfaces at caesarean section--a systematic review. AB - BACKGROUND: Caesarean section (CS) is a very common surgical procedure worldwide. Suturing the peritoneal layers at CS may or may not confer benefit, hence the need to evaluate whether this step should be omitted or not. OBJECTIVES: To assess the effects of non-closure as an alternative to closure of the peritoneum at CS on intraoperative, immediate and later postoperative, and long-term outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group Trials Register (November 2002) and the Cochrane Central Controlled Trials Register (October 2003). SELECTION CRITERIA: Randomised controlled trials that compared leaving the visceral and/or parietal peritoneum unsutured at CS with suturing the peritoneum, in women undergoing elective or emergency CS. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers. MAIN RESULTS: Nine trials involving 1811 women were included and analysed. The methodological quality of the trials was variable. Non-closure of the peritoneum reduced operating time when both layers or one layer was not sutured. For both layers, the operating time was reduced by 7.33 minutes (95% confidence interval (CI): -8.43 - -6.24). There was significantly less postoperative fever and reduced postoperative stay in hospital for non-closure of the visceral peritoneum and non-closure of both layers. There were no other statistically significant differences. The trend for analgesia requirement and wound infection tended to favour non-closure, while endometritis results were variable. Long-term follow-up in 1 trial showed no significant differences. The power of the latter study to show differences was low. CONCLUSIONS: There was improved short-term postoperative outcome if the peritoneum was not closed. Long term studies following CS are limited, but data from other surgical procedures are reassuring. At present there is no evidence to justify the time taken and cost of peritoneal closure. PMID- 15751209 TI - [Recommendations of the Polish Society of Obstetrics and Gynecology regarding pathology of early pregnancy after in vitro fertilization]. PMID- 15751211 TI - [Placenta previa accreta]. AB - OBJECTIVES: The purpose of our study was to assess the relationship between previous cesarean section and placenta previa accreta and to estimate the incidence of placenta accreta et previa accreta as the indication for peripartum hysterectomy. MATERIALS AND METHODS: The records of all patients delivered with the diagnosis of placenta previa accreta during the period from 1992-2002 at Hospital in Chojnice were reviewed. Statistical analyses were carried out to determine the relationship between previous cesarean section and subsequent development of placenta previa accreta. We conducted a retrospective analysis of indications for peripartum hysterectomy. RESULTS: From a total 28,177 women, who delivered at the Chojnice Hospital, 15(0.05%) patients had placenta accreta, 63(0.2%) placenta previa. Among placenta previa deliveries 22(34.9%) patients had previous cesarean section. Out of 15 patients with placenta accreta 10(66.7%) had placenta previa. Incidence of placenta accreta per case of placenta previa was 158.7 per 1000. The incidence of placenta previa accreta significantly increased in those with previous post cesarean scars. This incidence increased as the number of previous cesarean sections increased. The most common indication for peripartum hysterectomy was placenta accreta--48.4%, incidence of placenta previa accreta was accounts for 32.3% of all indications. CONCLUSIONS: The association between placenta previa accreta and prior cesarean section was confirmed. The incidence of placenta accreta increased as the number of previous cesarean sections increased. Patients with an antepartum diagnosis of placenta previa, who have had a previous cesarean section should be considered at high risk for developing placenta accreta. The most common indication for peripartum hysterectomy in this study was placenta previa accreta. PMID- 15751210 TI - [Ultrasound evaluation of NO-donor (L-arginine) therapy for intrauterine growth retardation]. AB - There are many methods of the growth restriction therapy, but the results are still not satisfactory. Actually, we are conducting a project which main conception is based on the use of the NO donor (L-arginine) in growth retardation therapy. This amino acid is an effective NO donor and has a vasodilating effect on the vessels and anti-aggregation effect on platelets, which plays a great role in improving feto-maternal blood flow. The main aim of this paper is the ultrasound evaluation of the effectiveness of NO-donor (L-arginine) therapy for intrauterine growth retardation based on biometrical measurements (estimated fetal weight) compared with newborns weight estimation. METHODS: The investigated group comprised two groups of pregnant women with ultrasonographically diagnosed intrauterine growth retardation (biometry <10th percentile for the gestation age): --78 patients were treated by L-arginine 3 g daily p.o. in 20 days period. -30 patients not treated (no agreement for therapy) as controls. RESULTS: Ultrasound estimation of fetal weight at the start and at the end of the treatment showed a mean value increase of 642 g (SE 90g) using Shepard method and 648g (SE 94g) using Hadlock method respectively. Comparing not treated group there was assessed a mean value increase of 395 g (SE 77 g) using Shepard method and 404g (SE 82g) using Hadlock method respectively. There is statistical significant difference comparing an estimated fetal weight in both methods: p = 0.008 for Shepard calculation and p = 0.012 for Hadlock calculation. There was analyzed newborn weight also--in a treated group mean value was at 2823 g and in not treated group mean value was at 2495 g. There was no significant difference found but a positive tendency for treated group newborns weight is visible. In the treated group number of the growth retarded newborns was at 29% and in the not treated group was at 73%. There was found a significant difference comparing both newborns groups at p < 0.05. CONCLUSIONS: The ultrasound evaluation of the estimated fetal weight and after-birth weight estimation of the newborns showed an increased dynamic of the fetal development in the L-arginine treated group of pregnant women comparing to the not treated group. The ultrasound evaluation of the estimated fetal weight is a good diagnostic tool for properly monitoring the effectiveness of treatment with L-arginine for growth retarded fetuses. PMID- 15751212 TI - [Comparison of indications for cesarean section in 1985-86 and 2000-01. Analysis of changes]. AB - The aim of this research was a retrospective analysis of indications for caesarean section during last fifteen years and assessing recent trends. MATERIALS AND METHOD: 1106 pregnancies and deliveries by caesarean section at the Ist Clinic of Obstetrics and Gynaecology, Medical University of Warsaw in 1985-86 (547--I group) and 2000-01 (559--II group) were enrolled in the study. It was performed in order to estimate and evaluate such factors as age of women, mean duration of pregnancy, indications for caesarean section, complications, and general newborn condition. RESULTS: In 1985-86 and 2000-01 fetal distress was the most frequent indication for delivery via caesarean section (14.35% and 18.57% respectively, p=ns). It was followed by cephalopelvic disproportion (13.99%) and malposition (13.99%) in I group and by pregnant women diseases (14.07%) and malposition (12.45%) in II group. Analysing pathology of the placenta, we established that index of placenta praevia decreased from 2.49% to 0.57% (p < 0.01) and index of abruption of placenta from 2.25% to 0.81% (p = 0.05). Ratio of cases of cephalopelvic disproportion decreased statistically from 13.99% in the I group to 8.76% in the II group (p = 0.006). Index of prolonged labor in the I stage increased from 8.3% to 12.34% (p = 0.027). CONCLUSIONS: (1) Caesarean section is the surgical method of delivery for women with high risk pregnancy and complicated labor to protect mother and the foetus from the anticipated complications of delivery through natural passages. (2) The overall rate of caesarean section has risen because of widening of the indications. (3) Fetal distress has been the most frequent indication for caesarean section in last fifteen years. (4) Index of indications for cesarean section other than obstetric -ophthalmic, cardiological, neurological and orthopaedic, increased as a result of better health care of pregnant women. (5) Decision to perform caesarean section should be taken in consideration of present clinical situation and future consequences of abdominal delivery. PMID- 15751213 TI - [Frequency of vaginal infections in pregnant women in the Department of Perinatology and Gynaecology in Zabrze]. AB - OBJECTIVES: The analysis of vaginal infections in pregnant women admitted to the Department of Perinatology and Gynaecology in Zabrze, from January 2001 to September 2003. MATERIALS AND METHODS: They were screened for BV, candidiasis and trichomoniasis. 450 pregnant women (the age of pregnancy 25 +/- 9 weeks gestation) were examined for the presence of these microorganisms in the vagina. Diagnosis was based on symptoms, pH measurement and microscopic examination of the vaginal discharge. CONCLUSIONS: The most common vaginal infection was candidiasis--188 (42%), 86 (19 %) women had BV, 17 (4%) Trichomonas vaginalis and 159 (35%) patients had normal vaginal flora. PMID- 15751214 TI - [Effectiveness of surgical treatment of patients with stage IA endometrial cancer]. AB - OBJECTIVES: The aim of the study was to analyse results of surgical treatment patients with stage IA-G1, G2 endometrial cancer treated in Center of Oncology in Krakow between 1985 and 1997. MATERIALS AND METHODS: The research included a group of 44 women. All patients underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH and BSO) without complementary treatment. RESULTS: Five NED survival was 95.5%. During the 5-year follow-up period 2 patients died, I of myocardial infarction and 1 of cerebral hemorrhage. CONCLUSION: Total abdominal hysterectomy and bilateral salpingo-oophorectomy are the treatment of choice for stage IA-G1, G2 endometrial cancer. PMID- 15751215 TI - [Laparoscopic supravaginal hysterectomy--in our own experience]. AB - INTRODUCTION: The role of laparoscopic supravaginal hysterectomy has increased a minimal invasive surgery. The reason for this is that the intact pericervical ring and neurovascular supply play an important role in pelvic floor support. Also, women's health awareness has increased. OBJECTIVES: The objective of this article was a retrospective analysis of 50 laparoscopic supracervical hysterectomies and advantages of this new surgical method. MATERIALS AND METHODS/DESIGN: These surgical procedures were performed in Sw. Rodziny Hospital and Gynecology of Obstetric in Warsaw in years 2002-2003. The duration of surgical procedure, complications, and duration of hospitalization were evaluated on the basis of retrospective patients case histories. RESULTS: The medium duration of surgical procedure was 95 minutes, the medium duration of hospitalization after the surgical procedure was 3,3 days. There were not any severe complications. CONCLUSIONS: At the end it was stated that laparoscopic supravaginal hysterectomy is minimal invasive surgical technique, which can be recommended if vaginal hysterectomy is contraindicated and there are no indications to cervix excision. PMID- 15751216 TI - [The effect of selected sperm characteristics on their fertilizing potential during IVF procedures]. AB - OBJECTIVE: Basic seminological criteria were analyzed to assess a percentage of fertilized oocytes during classical IVF procedure. DESIGN: A prospective study. MATERIALS AND METHODS: One hundred fifty semen samples from patients undergoing IVF were studied. Sperm morphology in raw semen and after its selection on a density gradient (6 structural defects of a gamete were taken into consideration), sperm concentration and motility parameters according to WHO criteria were analyzed. RESULTS: Patients achieving fertilization had a significantly higher median proportion of normal forms, concentration of sperm, motility grade A, B, C and significantly lower median values for head defects, acrosomal defects, and motility grade D, than those who did not have fertilization. A marked improvement of sperm morphology after density gradient selection was also noted. Apart from significant correlations obtained for selected semen parameters with proportion of fertilized eggs, logistic regression analysis showed that the predictive value for the IVF oocyte fertilization, using normal morphology of sperm before and after selection, grade A, B and C of sperm movement in a raw semen accounted for 62.4% of fertilization. CONCLUSION: It seems that to obtain a better efficacy of IVF prognosis, functional tests have to be added to routine semen parameters. PMID- 15751217 TI - [Repeated torsion of the normal adnexa in a premenarchal girl]. AB - Recurrent normal adnexal torsion is rare. The clinical presentation is poor and nonspecific. Authors present 11-years old girl with repeated torsion of the normal adnexa. At the first laparotomy salpingo-oophorectomy was performed. Next two incidences were treated conservatively by ovary detorsion in laparoscopy. After second laparoscopy the girl was referred to the Adolescent Gynecology Outpatient Clinic. Preventive oophoroplexy was performed. After half of a year observation the girl reported no complaints and no abnormalities in physical examinations were found. Oophoroplexy in the cases of repeated normal adnexal torsion is a very useful method of prevention of recurrence. PMID- 15751218 TI - [Uretero-uterine fistula as complication of the cesarean section]. AB - CASE PRESENTATION: 26 y.o woman 3 weeks after cesarean section complained of urine leakage from vagina. Urography revealed uretero-uterine fistula. Surgery was performed--3 dexon sutures attaching strictured ureter to the uterine were localized and then cut off. Distally obliterated ureter was separated out of adjacent tissues. The strictured part of ureter was then incised on the length of ca 8 cm, pig tail catheter was induced and human dura mater allograft was put on and sewed in into the strictured segment. In the long term follow-up the woman gave birth to two more children in the vaginal delivery without any complications. Urographies revealed no fistula, no stricture recurrency and normal urine passage. PMID- 15751219 TI - [Examination of the RHD gene by non-invasive method in plasma of alloimmunized mother allowed to abandon invasive procedures and to deliver a healthy child]. AB - RHD gen of the fetus was examined by a non-invasive method in pregnant woman with a high titre of anti-D antibody, developed during previous pregnancy, which resulted in haemolytic disease of the newborn. For RHD gene evaluation, the DNA from maternal plasma was examined, by the use of real-time PCR. RHD gene was not detected. Additional detection of gen GSTM1 in maternal plasma,which was present in the father but absent in the mother, assured us that we examined the fetal, not maternal RHD gene. Thus, the cordocentesis was not performed. The mother delivered healthy RhD negative daughter. PMID- 15751220 TI - [Delayed delivery in multiple gestations--a case report and review of the literature]. AB - The incidence of multiple pregnancies increases with concomitant increased risk in maternal and fetal complications. Delayed delivery is still a rare occurrence in the literature despite advances in reproductive medicine and increasing number of multiple gestations. Authors report a case of delayed delivery in a dichorionic, diamniotic twin pregnancy. A 28-year-old woman delivered her first twin in the 25th week of gestation, 4 weeks after the preterm rupture of membranes. The newborn died on the first day of life due to prematurity. Cervical cerclage was performed, patient received antibiotics, corticosteroids and tocolytics. The second twin was successfully delivered by cesarean section in the 28th week of pregnancy, 18 days after the first twin was born. The baby was discharged home in good condition on the 64th day after the delivery. The conclusion was, that using tocolysis, antenatal steroids to stimulate lung maturation, antibiotics and cervical cerclage after delivery of the first fetus we can improve the life-expectancy of the retained fetus. Delayed delivery is a reasonable option for patients with multifetal pregnancies and premature delivery of the first baby or preterm rupture of membranes in the second trimester. PMID- 15751222 TI - [Some remarks about lichen sclerosus vulvae]. AB - This paper describes the different points of view on the pathogenesis of lichen sclerosus vulvae, the relationship to cancer and the methods of treatment. PMID- 15751221 TI - [Placenta accreta: epidemiology, molecular mechanism (hypothesis) and some clinical remarks]. AB - OBJECTIVE: To develop the hypothesis on biochemical mechanism of the placenta accreta and to review the epidemiological information on that complication. STUDY DESIGN: Data collected in 1995-2002 in the Medline System were the main source of analyzed literature. RESULTS: Placenta accreta occurs in approximately 1 of 2500 deliveries, however among women with placenta praevia, the incidence is nearly 10%. Independent risk factors for placenta accreta are previous cesarean delivery and maternal age > or =35 years. Postpartum hemorrhage is the main cause of maternal mortality. Hysterectomy is often performed to save the life of the mother. Ligation of internal pelvic arteries is considered to be ineffective procedure in about 50%. An another option in management of hemorrhage is embolisation of pelvic arteries. Recombinant factor VIIa is recommended is a new agent in pharmacological therapy. Our hypothesis is as follows: The complex of uPA/uPAR (urokinase plasminogen activator/receptor of urokinase plasminogen activator complex) plays crucial role in the generation of plasmin-dependent proteolysis, which takes place in the surface of trophoblast. Penetration of the villi into the tissues is controlled by plasminogen activator inhibitors- plasminogen activator inhibitor 1 (PAI-1) and plasminogen activator inhibitor 2 (PAI-2), mainly by PAI-2. PAI-2 inactivates uPA/uPAR complexes forming triplicate complexes. If the PAI-2 concentration in placenta and myometrium is low, the invasion of placenta villi is excessive. CONCLUSION: Disturbance of balance between plasminogen activator and their inhibitors (PAI-1 and PAI-2) in placenta and myometrium can lead to the formation of placenta accreta. PMID- 15751223 TI - Dysregulation of the hypothalamic-pituitary-gonadal axis with menopause and andropause promotes neurodegenerative senescence. AB - Senescence is characterized neurologically by a decline in cognitive function, which we propose is the result of degenerative processes initiated by the dysregulation of the hypothalamic-pituitary-gonadal (HPG) axis with menopause and andropause. Compelling epidemiologic evidence to support this assertion includes the increased prevalence of Alzheimer disease (AD) in women, the correlation of serum HPG hormones with disease and the decreased incidence, and delay in the onset of AD following hormone replacement therapy. Dysregulation of the axis at this time leads to alterations in the concentrations of all serum HPG hormones (decreased neuronal sex steroid signaling, but increased neuronal gonadotropin releasing hormone, luteinizing hormone, and activin signaling). Hormones of the HPG axis, receptors for which are present in the adult brain, are important regulators of cell proliferation and differentiation during growth and development. Based on this, we propose that dysregulated HPG hormone signaling with menopause/andropause leads to the abortive reentry of differentiated neurons into the cell cycle via a process we term "dyosis." Interestingly, the major biochemical and neuropathologic changes reported for the AD brain also are intimately associated with neuron division: altered AbetaPP metabolism, Abeta deposition, tau phosphorylation, mitochondrial alterations, chromosomal replication, synapse loss, and death of differentiated neurons. Recent evidence supports the premise that AD-related biochemical changes are likely the combined result of increased mitotic signaling by gonadotropins and GnRH, decreased differentiative and neuroprotective signaling via sex steroids, and increased differentiative signaling via activins. This results in a hormonal milieu that is permissive of cell cycle reentry but does not allow completion of metaphase. Partial resetting of the axis following administration of normal endogenous sex steroids delays the onset and decreases the incidence of AD. Ideally, supplementation with HPG hormones should mimic closely the serum concentrations of all HPG hormones in reproductive men and cycling women to prevent dyotic signaling and attempted neuron division. PMID- 15751224 TI - The role of caspase cleavage of tau in Alzheimer disease neuropathology. AB - Alzheimer disease (AD) is characterized by the accumulation of amyloid plaques and neurofibrillary tangles within selective brain regions. In addition, cell death pathways become active leading to neurodegeneration. Caspase activation, a key step in the programmed cell death pathway known as apoptosis, occurs in AD and leads to the proteolytic cleavage of several neuronal proteins. Previously, it was hypothesized that the development of the classical hallmarks of AD, amyloid plaques and neurofibrillary tangles, occur independently and do not involve the activation of caspases. However, recent studies suggest that plaques, tangles, and caspase activation share a common pathway. Beta-amyloid, the main component of amyloid plaques, activates caspases. Activated caspases can in turn cleave tau, the main component of neurofibrillary tangles. Caspase-cleaved tau (deltatau) may initiate or accelerate the development of tangle pathology. Tau, when cleaved by caspases at Asp421, "seeds" filamentous aggregates in vitro. Caspase-cleaved tau also adopts the MC1 conformation, one of the earliest pathologic events in tangle formation. Importantly, deltatau occurs early in the development of tangle pathology within AD brains and in a transgenic mouse model of AD. This review summarizes recent evidence suggesting that caspase cleavage of tau plays an important role in the development of neurofibrillary tangle pathology. In addition, a model is presented whereby caspase cleavage of tau provides a mechanistic link between the development of amyloid and tangle pathologies. PMID- 15751225 TI - Extensive involvement of autophagy in Alzheimer disease: an immuno-electron microscopy study. AB - The accumulation of lysosomes and their hydrolases within neurons is a well established neuropathologic feature of Alzheimer disease (AD). Here we show that lysosomal pathology in AD brain involves extensive alterations of macroautophagy, an inducible pathway for the turnover of intracellular constituents, including organelles. Using immunogold labeling with compartmental markers and electron microscopy on neocortical biopsies from AD brain, we unequivocally identified autophagosomes and other prelysosomal autophagic vacuoles (AVs), which were morphologically and biochemically similar to AVs highly purified from mouse liver. AVs were uncommon in brains devoid of AD pathology but were abundant in AD brains particularly, within neuritic processes, including synaptic terminals. In dystrophic neurites, autophagosomes, multivesicular bodies, multilamellar bodies, and cathepsin-containing autophagolysosomes were the predominant organelles and accumulated in large numbers. These compartments were distinguishable from lysosomes and lysosomal dense bodies, previously shown also to be abundant in dystrophic neurites. Autophagy was evident in the perikarya of affected neurons, particularly in those with neurofibrillary pathology where it was associated with a relative depletion of mitochondria and other organelles. These observations provide the first evidence that macroautophagy is extensively involved in the neurodegenerative/regenerative process in AD. The striking accumulations of immature AV forms in dystrophic neurites suggest that the transport of AVs and their maturation to lysosomes may be impaired, thereby impeding the suspected neuroprotective functions of autophagy. PMID- 15751226 TI - Mitochondrial myopathy and rhabdomyolysis associated with a novel nonsense mutation in the gene encoding cytochrome c oxidase subunit I. AB - Mitochondrial DNA (mtDNA) mutations associated with rhabdomyolysis are rare but have been described in sporadic cases with mutations in the cytochrome b and cytochrome c oxidase (COX) genes and in 3 cases with tRNALeu mutation. We report a novel heteroplasmic G6708A nonsense mutation in the mtDNA COI gene encoding COX subunit I in a 30-year-old woman with muscle weakness, pain, fatigue, and one episode of rhabdomyolysis. Histochemical examination of muscle biopsy specimens revealed reduced COX activity in the majority of the muscle fibers (approximately 90%) and frequent ragged red fibers. Biochemical analysis showed a marked and isolated COX deficiency. Analysis of DNA extracted from single fibers revealed higher levels of the mutation in COX-deficient fibers (> 95%) compared with COX positive fibers (1%-80%). The mutation was not detected in a skin biopsy, cultured myoblasts, or blood leukocytes. Nor was it identified in blood leukocytes from the asymptomatic mother, indicating a de novo mutation that arose after germ layer differentiation. Western blot analysis and immunohistochemical staining revealed that reduced levels of COX subunit I were accompanied by reduced levels of other mtDNA encoded subunits, as well as nuclear DNA encoded subunit IV, supporting the concept that COX subunit I is essential for the assembly of complex IV in the respiratory chain. PMID- 15751227 TI - Nogo-A and nogo receptor expression in demyelinating lesions of multiple sclerosis. AB - A myelin-associated neurite outgrowth inhibitor, Nogo-A, plays a key role in inhibition of axonal regeneration following injury and ischemia in the central nervous system (CNS). Because axonal injury is a pathologic hallmark of multiple sclerosis (MS), we have investigated the expression of Nogo-A and its receptor NgR in four MS and 12 non-MS control brains by immunohistochemistry. Nogo-A expression was markedly upregulated in surviving oligodendrocytes at the edge of chronic active demyelinating lesions of MS and ischemic lesions of acute and old cerebral infarction, whereas NgR expression was greatly enhanced in reactive astrocytes and microglia/macrophages in these lesions when compared with their expression in the brains of neurologically normal controls. Nogo-A and NgR were also identified in a subpopulation of neurons. In contrast, Nogo-A was undetectable in reactive astrocytes and microglia/macrophages and NgR was not expressed on oligodendrocytes in any cases examined. Western blot analysis and double labeling immunocytochemistry identified the constitutive expression of NgR in cultured human astrocytes. These results suggest that Nogo-A expressed on oligodendrocytes might interact with NgR presented by reactive astrocytes and microglia/macrophages in active demyelinating lesions of MS, although biologic effects caused by Nogo-A/NgR interaction among glial cells remain unknown. PMID- 15751229 TI - The pathology of multiple sclerosis is location-dependent: no significant complement activation is detected in purely cortical lesions. AB - Complement activation is known to occur in white matter multiple sclerosis (MS) lesions. It is thought to mediate oligodendrocyte/myelin damage and to be a marker of pathologic heterogeneity among individuals. Less is known about complement deposition in the gray matter in MS. The aim of this study was to characterize the presence and distribution of complement activation products in cortical MS lesions. Immunohistochemical staining was performed on cryostat sections from the brains of 22 MS patients and 5 nonneurologic control patients obtained at autopsy. Deposition of the complement activation products C1q, C3d, and C5b-9 (membrane attack complex) was detected on and within macrophages/microglia and astrocytes and in blood vessel walls in white matter MS lesions. C3d and C4d were detected along myelin sheaths at the edge of the lesions. In the gray matter part of combined gray matter/white matter lesions complement activation was less frequent, but increased immunopositivity was detected for C3d on blood vessels, and for C3d and C4d on myelin at the border of lesions, when compared with control areas. In contrast, in the purely cortical lesions, the extent of complement deposition in general was low. In conclusion, the role of complement in MS pathogenesis seems lesion location-dependent. PMID- 15751228 TI - Effects of chronic glucocorticoid administration on insulin-degrading enzyme and amyloid-beta peptide in the aged macaque. AB - Insulin-degrading enzyme (IDE) has been identified as a candidate protease in the clearance of amyloid-delta (Abeta) peptides from the brain. IDE activity and binding to insulin are known to be inhibited by glucocorticoids in vitro. In Alzheimer disease (AD), both a decrease in IDE levels and an increase in peripheral glucocorticoid levels have been documented. Our study investigated the effects of glucocorticoid treatment on IDE expression in vivo in 12 nonhuman primates (Macaca nemestrina). Year-long, high-dose exposure to the glucocorticoid cortisol (hydrocortisone acetate) was associated with reduced IDE protein levels in the inferior frontal cortex and reduced IDE mRNA levels in the dentate gyrus of the hippocampus. We assessed Abeta40 and Abeta42 levels by ELISA in the brain and in plasma, total plaque burden by immunohistochemistry, and relative Abeta1 40 and Abeta1-42 levels in the brain by mass spectrometry. Glucocorticoid treatment increased Abeta42 relative to Abeta40 levels without a change in overall plaque burden within the brain, while Abeta42 levels were decreased in plasma. These findings support the notion that glucocorticoids regulate IDE and provide a mechanism whereby increased glucocorticoid levels may contribute to AD pathology. PMID- 15751230 TI - Alpha-synuclein lesions in normal aging, Parkinson disease, and Alzheimer disease: evidence from the Baltimore Longitudinal Study of Aging (BLSA). AB - Alpha-synuclein (alpha-synuclein) lesions are characteristic of idiopathic Parkinson disease (PD) and other alpha-synucleinopathies. To study the frequency of alpha-synuclein lesions in normal aging and how frequently they coexist with lesions of Alzheimer disease (AD), we examined the autopsy brains from normal and demented subjects in the Baltimore Longitudinal Study of Aging (BLSA) (n = 117). We found that the overall frequency of alpha-synuclein lesions was 25%, with 100% in 7 cases of PD, 31.5% in 56 cases with AD lesions, and 8.3% among 36 older control brains. Among brains with AD lesions, the frequency of alpha-synuclein pathology was higher in those with higher scores for neuritic plaques, but not in those with higher scores for neurofibrillary tangles. Our observations indicate that alpha-synuclein lesions are uncommon in aged control subjects. Finally, the coexistence of Abeta amyloid and alpha-synuclein pathology in AD brains suggests that the pathogenic mechanism/s leading to the accumulation of Abeta and alpha synuclein may be similar. PMID- 15751231 TI - Proliferative potential of human astrocytes. AB - Although a number of studies have demonstrated proliferation of nonneoplastic astrocytes in experimental animal models, the proliferative potential of human astrocytes has not been well defined. Using double-label immunohistochemistry, we identified proliferating cells with the proliferation marker MIB-1 and astrocytes with glial fibrillary acidic protein staining in human biopsy and autopsy tissue. MIB-1 labeling of astrocytes was monitored in a variety of conditions containing significant numbers of reactive astrocytes, including infections, arteriovenous malformations, demyelinating lesions, metastatic tumors, and long-standing gliosis. Twenty-nine of a total of 54 cases showed no evidence of astrocyte specific MIB-1 labeling despite prominent reactive changes. An average proliferation rate of 0.9% was present in the remaining 25 cases. Labeling indices were highest in infectious conditions and acute demyelinating lesions. We also examined astrocyte proliferation in 5 cases of progressive multifocal leukoencephalopathy. Astrocytic labeling indices were notably elevated in these cases, with an average labeling index of 5.8%. We conclude that low, but appreciable, astrocytic proliferation may occur in nonneoplastic human astrocytes. These findings have implications for astrocyte function in the normal and disease states and for the diagnostic distinction between reactive lesions and low-grade astrocytic neoplasms. PMID- 15751232 TI - Bioptics by angle-supported phakic lenses and photorefractive keratectomy. AB - PURPOSE: To assess efficacy and safety of the combination of angle-supported phakic intraocular lenses (IOLs) and photorefractive keratectomy (PRK) for the correction of myopia and astigmatism. METHODS: Prospective, non-randomized single surgeon study on 48 patients (76 eyes) undergoing angle-supported phakic IOL implantation with surgical peripheral iridectomy, followed 2 to 3 months later by PRK to correct residual refractive error. Twenty-three patients (33 eyes) achieved good uncorrected visual acuity with IOL implantation alone and did not undergo PRK. Thus, the study was completed by 25 patients (43 eyes) with preoperative mean defocus equivalent (DEQ) of 15.73 D (SD 4.67 D) and mean astigmatism of -2.87 D (SD 1.39 D). RESULTS: Eight months after PRK, mean spherical equivalent was -0.08. Mean DEQ was 0.47 D (SD 0.37); 42/43 eyes (98%) were within +/-1 D of DEQ, and 33/43 eyes (77%) within +/-0.5 D. Mean uncorrected visual acuity was 0.7 (SD 1.9). Safety index was 1.25; efficacy index 1.11. Best corrected visual acuity improvement (0.16) was statistically significant (95% CI: 1.1 to 2). Halos were moderate in 6/25 patients (24%); severe in 1/25 patients (4%). Endothelial cell density decreased by -6.6%. Pain after PRK was severe in 3/25 patients (12%) and moderate in 13/25 patients (52%). Complications were recurrent iridocyclitis in one eye, transient ocular hypertension in two eyes, and incomplete iridectomy in one eye. CONCLUSIONS: Angle-supported phakic IOLs followed by adjustment by PRK offer good efficacy, predictability, and safety to manage large refractive myopic errors. PMID- 15751233 TI - Comparison of keratometric and topographic cylinder and axis measurements on normal corneas with low astigmatism. AB - PURPOSE: To evaluate agreement in measurements of astigmatic axis power and location between keratometry and computer assisted videokeratography (corneal topography) on normal corneas with less than 1.50 D of idiopathic astigmatism. METHODS: Keratometric readings with the 10 SL/O Zeiss ophthalmometer and corneal topographic maps with the TMS-1 were obtained by two independent examiners on 32 normal corneas. Measurement agreement between the two instruments was evaluated in regard to steep and flat meridian power and location, and in astigmatism magnitude (D). RESULTS: The limits of agreement (d-2 SD to d+2 SD) between the two instruments were found to be broad for clinical purposes in measuring the steep meridian power (-0.16 to -1.20 D), flat meridian power (0.43 to -1.25 D), and astigmatism (0.60 to -1.12 D). A constant bias of the TMS-1 towards the 10 SL/O Zeiss ophthalmometer was found, in measuring steeper both principal meridians and higher amount of astigmatism. Mean location difference was 19 degrees (+/-190) for the steep meridian and 17 degrees (+/-20 degrees) for the flat meridian. CONCLUSIONS: Despite the differences seen in measurements between the 10 SL/O ophthalmometer and the TMS-1, these differences may be clinically small enough for the methods to be used interchangeably in measuring only the magnitude of astigmatism on normal corneas. However, the disagreement in astigmatism axes is too great to be ignored. PMID- 15751234 TI - Epithelial cell, keratocyte, and endothelial cell apoptosis in Fuchs' dystrophy and in pseudophakic bullous keratopathy. AB - PURPOSE: To elucidate the pathomechanism of Fuchs' dystrophy and pseudophakic bullous keratopathy (PBK) by examining cell apoptosis in different corneal layers. METHODS: The authors studied corneal buttons obtained from 21 eyes following central penetrating keratoplasty: 14 corneal buttons (13 patients, age 70.8+/-10.0 years) with Fuchs' dystrophy, and 7 buttons (7 patients, age 69.6+/ 10.2 years) with PBK. Four buttons from enucleated eyes with choroidal melanoma served as controls. Histologic changes were examined using light microscopy with hematoxylin-eosin (HE) staining. The average numbers of apoptotic cells per field of view (125x magnification) in separate samples of the epithelial, stromal, and endothelial layers were determined using the TUNEL (terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling) assay. RESULTS: In 11 of the Fuchs' dystrophy corneas and 2 of the PBK corneas, apoptotic activity was detected. In the control corneas no apoptotic activity was found. Compared to the controls there was a statistically significant difference in the mean (normalized) apoptotic cell numbers for all three layers (p=0.01 in each case) in the Fuchs' dystrophy corneas, and for the stromal layer (p<0.01) in PBK corneas. The apoptotic cell numbers for the epithelial and endothelial layers of the latter were higher, but the difference was not statistically significant (p=0.07, 0.07). CONCLUSIONS: Apoptosis may play a role in the pathomechanism of Fuchs' dystrophy and in keratocyte death in corneas with PBK. PMID- 15751235 TI - Infectious keratitis in South Australia: emerging resistance to cephazolin. AB - PURPOSE: To analyze the microbiologic spectrum and patterns of resistance of infectious keratitis in patients treated at a tertiary hospital in South Australia. METHODS: Retrospective review of microbiology laboratory records of all patients with infectious keratitis who had corneal scrapings, from 1998 to 2003. All records were subsequently reviewed for Gram staining and culture results, as well as antibiotic sensitivity and resistance. RESULTS: Positive corneal cultures were obtained in 134 out of 211 patients who had corneal scrapings (63.5%). Coagulase negative Staphylococcus was the commonest pathogen identified (29.8% of positive cultures), followed by Staphylococcus aureus (18.7%), Pseudomonas aeruginosa (12.7%), Moraxella (6.7%), Streptococcus pneumonia (6.0%), and fungal keratitis (5.2%). In 43.3% of culture positive cases, the organisms were also identified in Gram stain, and in all these cases there was a full correlation between the two methods. In vitro sensitivities were highest for gentamicin. Fourteen cases (35%) of coagulase negative Staphylococcus were found to be resistant to cephazolin. No ciprofloxacin resistance was identified in all Pseudomonas isolates tested. CONCLUSIONS: Staphylococcus species continue to be the commonest causative organism for infectious keratitis; however, there is an emerging resistance to cephazolin, which is commonly used as the first-line antibiotic for Gram-positive cocci. PMID- 15751236 TI - The effect of adjunctive Mitomycin C in Ahmed glaucoma valve implantation. AB - PURPOSE: To evaluate the effectiveness and safety of adjunctive mitomycin C (MMC) in Ahmed glaucoma valve (AGV) implantation for refractory glaucoma. METHODS: Twenty-two eyes of 22 patients who underwent AGV implantation with adjunctive MMC (0.5 mg/ml) for 3 minutes (Group A) were compared to a control group of 26 eyes of 26 patients (Group B) who received AGV implantation without MMC. Nine patients were female and 13 patients were male in Group A and 11 patients were female and 15 patients were male in Group B. The mean age was 56 years ranging from 13 to 77 in Group A and 58 ranging from 14 to 71 years in Group B. Success was defined as an intraocular pressure (IOP) between 4 and 21 mmHg with or without glaucoma medication and with no additional glaucoma surgery, phthisis, or loss of light perception. RESULTS: The probability of success at 1 year was 86.36% and 80.76% in Group A and Group B, respectively. The number of glaucoma medications decreased from 2.82 to 0.56 in Group A and from 2.65 to 0.73 in Group B. Postoperative hypotony as an early complication was higher in Group A (31.81% in Group A and 15.38% in Group B). As a late postoperative complication, tube exposure developed in 3 patients (13.63%) in Group A and no such complication was seen in Group B. There was no statistically significant difference in success and complication rates between the two groups (p>0.05). CONCLUSIONS: Although adjunctive MMC in AGV implantation is safe and effective, it may not offer a better chance of surgical success compared with AGV implantation without MMC. PMID- 15751237 TI - Knowledge base and preferred methods of obtaining knowledge of glaucoma patients. AB - PURPOSE: To gather information regarding patient's understanding of glaucoma and the manner in which patients wish to learn about the disease with the intent of improving patient education. METHODS: Forty-four of sixty randomly selected ophthalmologists (73%) asked four of their patients consecutively to complete a questionnaire about glaucoma. The selection of questions was based on focus group interviews and suggestions from several experts. Topics included knowledge about glaucoma and its treatment, the need for information, and preferred providers and methods of patient education. RESULTS: Fifty percent of the patients had 49% or less correct answers to questions about glaucoma or its treatment. Per item the correct answers ranged from 5% to 90%. Lack of knowledge was associated with low level of education, short duration of glaucoma, high age, and no preference for the Internet as method of supplying information. These variables, however, did not identify groups with a considerable lack of knowledge sufficiently accurately to target patient education. A high need for information was observed and included information about the patient's own glaucoma. Almost all patients preferred the ophthalmologist and many also a nurse or a representative of the Glaucoma Patient Society as providers of information. Written material was the preferred method. CONCLUSIONS: Patient education should address all patients. A patient education program should cover a wide range of topics with a focus on general information through written material and information tailored to the individual glaucoma patient's needs. The ophthalmologist is a key- person, but others could play an important role in patient education. PMID- 15751238 TI - Optic nerve damage in highly myopic eyes with chronic open-angle glaucoma. AB - PURPOSE: To compare the amount of optic nerve damage in relation to intraocular pressure in highly myopic eyes with chronic open-angle glaucoma versus non-highly myopic eyes with chronic open-angle glaucoma. METHODS: The comparative clinical observational study included 1841 eyes of 1100 patients with chronic open-angle glaucoma. The highly myopic study group consisted of 25 eyes with a myopic refractive error equal to or higher than -8 diopters. It was subdivided into eyes with an optic disc size larger than 2.7 mm2 and eyes with an optic disc smaller than 2.7 mm2. The control group included the remaining, non-highly myopic eyes (n=1816). For all patients, a morphometric analysis of color stereo optic disc photographs was performed. Main outcome measures were morphometric optic disc measurements and intraocular pressure. RESULTS: In the highly myopic, large-optic disc study group compared with the control group, maximal and minimal intraocular pressure readings were significantly (p<0.05) lower and neuroretinal rim area corrected for optic disc size was slightly (p=0.16) smaller. Comparing the total highly myopic study group with a control group adjusted for optic disc area, neuroretinal rim area was significantly (p=0.039) smaller in the study group with no significant difference in intraocular pressure measurements between the groups. CONCLUSIONS: At a given intraocular pressure in chronic open-angle glaucoma, optic nerve damage may be more pronounced in highly myopic eyes with large optic discs than in non-highly myopic eyes. This may suggest a higher susceptibility for glaucomatous optic nerve fiber loss in highly myopic eyes than in non-highly myopic eyes. PMID- 15751239 TI - Ocular blood flow evaluation in injured and healthy fellow eyes. AB - PURPOSE: To assess if injured eyes develop ocular blood flow disturbances that may contribute to development of traumatic glaucoma. METHODS: Twenty-five eyes of 25 patients hospitalized from January 1997 to July 1999 for blunt (15) or penetrating (10) eye injury and elevated intraocular pressure (IOP) (>23 mm Hg) were controlled at least 24 months after the trauma and underwent visual field examination, pulsatile ocular blood flow (pOBF), and color Doppler imaging (CDI) analysis of ophthalmic artery, central retinal artery, nasal and temporal short posterior ciliary arteries. Uninjured healthy eye was used as control. RESULTS: IOP was significantly higher in injured eyes (15.1+/-3.3 vs 13.0+/-2.7 mmHg; p<0.01), but only 2 eyes (8%) were under medical treatment. pOBF values were significantly lower in injured eyes: 11.25+/-6.56 microL/sec in the trauma eyes and 15.40+/-7.29 in fellow eyes (p=0.002). Resistivity index of all investigated retrobulbar vessels was very significantly higher in injured eyes than in fellow eyes (p<0.0001). There is no significant correlation between IOP and ocular blood flow disturbance. CONCLUSIONS: Long-term follow-up (mean 39+/-12 months) of injured eyes shows, besides a slight but significant increase of IOP, a very significant impairment of ocular blood supply to injured eyes compared to healthy fellow eyes with reduction of pulsatile ocular blood flow and marked increase of resistance to flow in all retrobulbar vessels. These anomalies may be considered an independent risk factor to develop traumatic glaucoma. PMID- 15751240 TI - Myogenic response reduction by high blood glucose levels in human retinal arterioles. AB - PURPOSE: The effect of elevated blood glucose level on the myogenic response of human retinal arterioles to acute increases in blood pressure is investigated. METHODS: The vascular response to raised blood pressure (Bayliss effect) was measured in 12 healthy volunteers by use of the retinal vessel analyzer (RVA). For a 9-minute period an on-line measurement of the diameter of a retinal branch arteriole was performed. After the first 3 minutes (baseline measurement) a second phase with 3 minutes of isometric exercise caused an acute rise in blood pressure, followed by 3 minutes of recovery (phase III). After the first session 100 g glucose were administered per os. After 30 minutes blood glucose was measured again and an identical second session was performed with higher blood glucose levels. The Wilcoxon test was used for statistical analysis. RESULTS: During the first session a rise in mean arterial pressure of 22.8 (+/-8.4) mmHg was followed by an arterial vasoconstriction of -6.6 (+/-1.7) %. The administration of 100 g glucose resulted in a significant rise in blood glucose levels within 30 minutes between the two sessions (4.35 mmol/L vs 7.46 mmol/L) (p=0.002). The blood pressure rise of 25.7 (+/-7.3) mm Hg in the second session was associated with a significant loss in arterial vasoconstriction of -2.3 (+/ 1.4) % (session I vs session II p=0.002). CONCLUSIONS: The myogenic response of the arterial wall in human retinal arterioles was significantly reduced during acute rise of blood glucose levels. PMID- 15751241 TI - Prevalence of blindness and visual impairment among Jordanian diabetics. AB - PURPOSE: To investigate the prevalence of blindness and visual impairment among a population of Jordanian diabetics. METHODS: A total of 986 diabetic patients were fully assessed, including complete history, examination, and laboratory tests. All patients underwent detailed eye examination, which included visual acuity, slit-lamp examination, tonometry, funduscopy, and fundus fluorescein angiography (FFA). RESULTS: Of all patients examined, 53.2% were male and 46.8% were female. The mean age and duration of diabetes were 55.3 and 11.9 years. Of all patients, 93.3% had type 2 while 6.7% had type 1 diabetes mellitus (DM). Over half (50.3%) were on oral hypoglycemic agents, 34% on insulin, and 14.5% on both types of treatment, whereas only 1.2% were on diet alone. The mean value for HbA1c was 7.7%. The prevalence of blindness among participants was found to be 7.4%, while 10.1% were visually impaired. Diabetic retinopathy (DR) was present in 64.1%, 37.8% had cataract, and 8.7% had undergone cataract surgery. Using multivariate logistic regression analysis, visual impairment was significantly associated with age, treatment of diabetes, and DR, while only age and retinopathy were significantly related to blindness. CONCLUSIONS: DM is a common disease in Jordan and DR is highly prevalent among Jordanian diabetics. National screening and educational programs are highly needed to reduce the risk of blindness and visual impairment among diabetic patients. PMID- 15751242 TI - Photodynamic therapy with verteporfin of choroidal neovascularization in angioid streaks: conventional versus early retreatment. AB - PURPOSE: To evaluate the effectiveness of conventional photodynamic therapy with verteporfin (PDT) in a series of patients with macular choroidal neovascularization (CNV) due to angioid streaks and to compare it to the effectiveness of early PDT retreatment. METHODS: This is a retrospective study of 24 eyes (22 consecutive patients) with subfoveal or juxtafoveal CNV secondary to angioid streaks treated with PDT from September 2000 through February 2003 and that completed at least the first year of follow-up. Until August 2001, retreatments were performed according to the conventional protocol for PDT every 3 months (Group 1, consisting of 11 eyes of 9 patients). After August 2001 (13 more eyes of 13 new patients), retreatments were performed earlier (every 8 weeks) when indicated (Group 2). The follow-up time ranged from 30 to 42 months and from 12 to 30 months in Groups 1 and 2. RESULTS: At the end of the follow-up, final best-corrected visual acuity decreased in 21 (87.5%), stabilized in 2 (8.3%), and improved in 1 (4.2%) of the total 24 eyes. In all, 19 of the 24 eyes (79.2%) had a final best-corrected visual acuity equal to or less than 20/400. There were not any statistically significant differences in final visual acuity between the two groups. CONCLUSIONS: In this large series of patients with macular CNV secondary to angioid streaks, the functional and the anatomic results of PDT were not satisfactory, even when retreatments were performed earlier than the conventional time of 3 months. PMID- 15751243 TI - Clinical outcome and subjective quality of life after photodynamic therapy in patients with age-related macular degeneration. AB - PURPOSE: Whereas the efficacy of photodynamic therapy (PDT) in preventing the progression of age-related macular degeneration (ARMD) is established, its effect on quality of life is under discussion. METHODS: All patients who underwent PDT during 2000 and 2001 at the University Eye Hospital of Mainz were interviewed using a standardized 82-item questionnaire on quality of life and patient satisfaction in ophthalmologic patients. Information was assessed in terms of 82 questions; global scores ranging from 1.0 (optimum self-estimated quality of life) to 4.0 (worst) were derived. Cataract patients' scores were used to characterize the ARMD patients' subjective outcome; the latter were then related to clinical outcome parameters via logistic regressions. RESULTS: A total of 84 patients (50% female, median age 77 years) were interviewed, who underwent a median of three PDT interventions. During the period of PDT treatment, their median decrease in visual acuity was 3 lines from 0.125 to 0.063. Patients who reported a subjective increase in visual function during this period showed a median private flexibility score of 1.86; patients with the subjective impression of visual function decrease, a median score of 2.71; the median scores for mobility were 2.00 versus 3.00, for flexibility in reading 1.91 versus 3.64, for psychological stress 1.56 versus 2.25, and for communicational flexibility 1.72 versus 2.25. The difference in reading flexibility was statistically significant (p=0.001) after correction for clinical cofactors. CONCLUSIONS: The established clinical benefit of PDT treatment concerning its efficacy in ARMD progression prevention coincides with an at least slight subjective benefit in quality of life and patient satisfaction. However, the latter is associated with the patients' subjective impression of visual acuity progression rather than with clinically validated outcome after PDT treatment. PMID- 15751244 TI - Central corneal thickness and intraocular pressure relationship in eyes with and without previous LASIK: comparison of Goldmann applanation tonometer with pneumatonometer. AB - PURPOSE: To investigate central corneal thickness (CCT) and intraocular pressure (IOP) relationship in eyes with and without previous corneal laser refractive surgery and to compare the estimates of two different tonometers-Goldmann applanation and pneumatonometer. METHODS: The study population included 234 glaucoma suspects who were referred to the glaucoma clinic with cup/disc ratios greater than 0.4, asymmetric cupping, and/or IOP greater than 22 mmHg during routine eye examination. Of those, 84 had previous myopic laser-assisted in situ keratomileusis (LASIK) (Group 1) while 150 of them did not (Group 2). CCT was measured by using optical coherence tomography (OCT) and IOP with both Goldmann applanation tonometer (GAT) and pneumatonometer (PT). In both groups, the difference between IOP estimates of two different tonometers and the relationship between CCT and IOP were analyzed. RESULTS: In eyes with previous LASIK, GAT measured IOP significantly lower than PT (mean difference of 3.8+/-1.9 mmHg, p<0.0001). In eyes with virgin corneas, IOP estimates of GAT or PT were not different from each other (19.9+/-2.8 versus 19.9+/-2.2 mmHg, respectively, p=0.81). In both groups, there was a significant positive correlation between CCT and IOP estimates of GAT(R=0.29, p=0.007 in eyes with LASIK and R=0.38, p<0.0001 in those without), while no similar relationship was present between CCT with those of PT (R=0.03, p=0.76 in eyes with LASIK and R=0.03, p=0.69 in those without). CONCLUSIONS: In eyes with previous LASIK, GAT measured IOP significantly lower than PT. Because IOP estimates of PT were found to be independent from CCT in all of the study eyes, this device was considered to be a more reliable method of IOP estimation than GAT in eyes with and without previous LASIK. PMID- 15751245 TI - Intravitreal triamcinolone acetonide in refractory pseudophakic cystoid macular edema: functional and anatomic results. AB - PURPOSE: To evaluate safety and efficacy of intravitreal triamcinolone acetonide (TAAC) injections in the treatment of refractory pseudophakic cystoid macular edema (CME). METHODS: Seven eyes of six patients (age range: 50-74) with pseudophakic CME resistant to standard treatment received intravitreal injections of 4 mg of TAAC with all vehicle. Mean preinjection duration of CME was 18.3 months. A mean of 2.1+/-1.2 (range 1 to 4) treatments were performed in four eyes (57.1 %) when visual acuity deteriorated towards baseline levels. Visual acuity assessment, optical coherence tomography (OCT), and fluorescein angiography (FFA) were performed pre- and postoperatively to evaluate results of TAAC injections. Intraocular pressure (IOP) and complications related to treatment were assessed. RESULTS: After 11.1+/-3.9 months, mean best-corrected visual acuity (BCVA) increased (p =0.019) from 20/132 to a best value of 20/38. Mean macular thickness decreased from 517.29+/-146.98 mm to a best value of 263.71+/-83.13 mm (p=0.0018). Area of fluorescein leakage decreased (p<0.0001) from 11.84+/-0.93 mm2 at baseline to a minimal value of 3.86+/-0.98 mm2. The anatomic and functional improvement appeared after 1 month from the intravitreal injection and persisted through at least 3 months of follow-up. At the end of follow-up BCVA, macular thickness, and area of fluorescein leakage did not differ from baseline. Four eyes (57.1 %) developed IOP values higher than 21 mmHg, controlled by topical treatment. Two patients developed an endophthalmitis-like reaction. CONCLUSIONS: Intravitreal TAAC was relatively safe and effective in resistant cases of pseudophakic CME with a temporary beneficial effect on visual acuity and macular edema. PMID- 15751246 TI - The efficacy of intravitreal triamcinolone acetonide on macular edema in branch retinal vein occlusion. AB - PURPOSE: To evaluate the effectiveness of intravitreal triamcinolone acetonide as primary treatment of macular edema in branch retinal vein occlusion. METHODS: Fifteen eyes of 15 patients with macular edema due to branch retinal vein occlusion (Group 1) who received 8 mg/0.2 ml of intravitreal triamcinolone injection as primary treatment were retrospectively evaluated. The control group (Group 2) consisted of 19 eyes of 19 patients who had received laser treatment for macular edema. The main outcome measures included best-corrected visual acuity, intraocular pressure, and macular edema map values of Heidelberg Retinal Tomograph II. RESULTS: In Group 1, mean visual acuity improved significantly from a mean logMAR (logarithm of minimal angle of resolution) value of 0.98+/-0.19 at baseline to a maximum of 0.24+/-0.24 during a mean follow-up time of 6.3 months. In the control group, the mean baseline log-MAR visual acuity before laser treatment was 1.02+/-0.22, and it was 0.50+/-0.28 at 6-month examinations. Mean improvement in visual acuity at 1-, 3-, and 6-month examinations was significantly higher in Group 1 when compared with the control group (for each, p<0.001). The mean edema map value of Group 1 significantly decreased by 40% at 6 month examinations when compared with preinjection value (p<0.001). In Group 1, mean increase in intraocular pressure elevation was 19.8% at the 1-month, 26.9% at 3-month, and 5.7% at 6-month visits, but intraocular pressures were under control with topical antiglaucomatous medications. CONCLUSIONS: Intravitreal triamcinolone acetonide injection may be a new and promising approach as initial therapy for macular edema due to branch retinal vein occlusion. PMID- 15751247 TI - Patient satisfaction and vision improvement after multiple surgery for recurrent retinal detachment. AB - PURPOSE: To assess patient satisfaction and functional status improvement after multiple surgery for recurrent retinal detachment (RRD) with proliferative vitreoretinopathy (PVR) grade C. Main outcome measures included visual acuity (VA), anatomic outcome, and patients' answers to a standardized multiple-choice questionnaire. METHODS: The authors retrospectively reviewed records of patients undergoing pars plana vitrectomy (PPV) for RRD with PVR grade C operated between 1997 and 1999. All included patients underwent a standardized telephone interview aimed at assessing the patients' visual performance and satisfaction. Statistical analysis used Wilcoxon signed-rank test, Mann-Whitney rank sum test, and Spearman rank correlation coefficient. p Values less than 0.05 were considered statistically significant. RESULTS: The population study included 62 patients with an average 19+/-4.1 months follow-up. The average number of operations was 2.9+/-0.7 with 1.45+/-0.50 recurrences. At the end of the study, 40/62 eyes had better than 5/400 vision and 14/62 better than 20/200; 2 patients had no light perception and 5 eyes still had RRD. Questionnaire answers yielded the following results: 61% believed their VA was better than before surgery, 13% the same, and 26% worse. Fifty-two percent noticed an improvement in binocular vision versus 35% stable and 13% worse: 84% thought their result had been worth the operation, 15% did not, and 2% did not know. Ninety percent believed the information they received before surgery was accurate. Results exceeded expectations in 35% of cases and matched them in 26%. Increase in binocular visual performance after intervention was significant for clothing, bathing, and home deambulation, climbing steps, watching TV, and reading, but not for car driving. The difference in VA improvement in satisfied and unsatisfied patients was significant. There was no significant correlation between patients' satisfaction and fellow-eye vision. CONCLUSIONS: The sample population showed a high rate of satisfaction and significant subjective improvement on four out of five tested activities, despite multiple surgeries and overall poor outcome, even in patients with a good VA in fellow eye. Many other factors such as visual field and contrast sensitivity improvement, not investigated by the authors, can play an important role in the visual functional status. Correct and extensive patient information remains critical in such settings. PMID- 15751248 TI - Macula off retinal detachments. How long can they wait before it is too late? AB - PURPOSE: To determine the association between the duration of macula off detachment and the visual outcome following corrective surgery. METHODS: Retrospective review of the medical records of patients who underwent surgery for macula off detachment over a 5 year period (April 1994- March 1999). RESULTS: There were 104 patients in the study. Patients with macula off detachments wait a mean of 2.6 weeks (+/-0.3 SE mean) before presentation and 1.8 weeks (+/-0.2 SE of mean) thereafter before surgery. The mean duration of detachment prior to surgical repair was 4.2 weeks (+/-0.3 SE mean). 78% of patients achieved a postoperative improvement in visual acuity. 36.5% achieved functional visual success of 6/12 at 3 months, which increased to 51% at final discharge. There was no significant difference in visual outcomes for patient undergoing internal vs external procedures (p=0.188). The preoperative visual acuity was the most significant predictor of post operative visual acuity (p<0.0005). Less than 40% of macula off detachments of > or =6 weeks duration will achieve a vision of 6/12 or better compared with 68.2% of patients with macula off detachments of < or =1 week. CONCLUSIONS: The best mean postoperative vision (LogMAR 0.35) was seen in patients with detachment of <1 week duration. Patients <60 years are more likely to achieve visual improvement despite the duration of the detachment. Macula off detachments of >6 weeks duration have a significantly poor postoperative visual prognosis. Awareness of this visual prognosis can assist in planning the timing of surgery to ensure an acceptable result. PMID- 15751249 TI - Ocular manifestations of congenital lamellar ichthyosis. AB - PURPOSE: To describe the ophthalmic manifestations in a series of children with congenital lamellar ichthyosis. These cases presented with varying types of eyelid abnormality associated with the systemic disease. The clinical features and ophthalmic management were studied. METHODS: The case histories of three children presenting to the oculoplastic clinic were reviewed. All were diagnosed with congenital lamellar ichthyosis and under the care of the Dermatology department. Family history and pedigree analysis was performed to determine mode of genetic inheritance. Ocular examination for visual acuity, eyelid and eyelash malposition, lid function and closure were carried out. Corneal examination including tests for exposure was also done. RESULTS: All three patients had eyelid position abnormalities from the systemic disease. There was no clinical evidence of conjunctival involvement. One patient required full thickness skin grafts to treat corneal exposure secondary to lower lid ectropion. One had mild lower lid ectropion but without corneal exposure. The third case had the unusual finding of inward turning of the anterior lamella of the upper eyelid with a marked lash ptosis and only mild ectropion of the lower lid. CONCLUSIONS: Congenital lamellar ichthyosis is a heterogeneous disorder with phenotypic variability. The most common eyelid abnormality is cicatricial ectropion of the upper and mainly lower eyelids. Most cases are managed conservatively although in severe cases secondary corneal exposure may require surgical correction. In this condition, to the best of our knowledge, the tendency for the eyelids to turn inwards has not previously been described. PMID- 15751250 TI - Bilateral complete avulsion of ocular globes in a Le Fort III maxillofacial fracture: a case report and review of the literature. AB - PURPOSE: The purpose of this paper is to demonstrate a case of bilateral complete avulsion of the globes following maxillofacial trauma. METHODS: A 23-year-old man with bilateral complete globe avulsion following a maxillofacial trauma. Both globes were luxated out of the orbit and suspended on the skin of the upper lid below the brows. No direct or indirect light reflexes or any eye movements could be noted. Computerized tomography showed complete lacerations of both optic nerves at a level just anterior to the optic canal. There were also multiple fractures corresponding to Le Fort III fracture with Le Fort II components. The brain parenchyma was normal with an exception of brain edema. As the globes were unsuitable for repositioning, both were enucleated. The maxillofacial fractures were immobilized with plates and screws. Although the patient gained consciousness with normal vital signs in the early postoperative period, he died on the ninth day due to Pseudomonas aeruginosa infection, despite invasive antibiotic treatment. DISCUSSION: The optic nerve and the globes are very resistant to mild and moderate trauma. The avulsion of the nerve at the canalicular or more posterior level may demonstrate central nervous system complications resulting in life-threatening conditions. The most critical issue in complete globe avulsions with a transected optic nerve is to rescue the vision. In total transected optic nerves the final eyes could only have had cosmetic benefits. In spite of promising experimental research on optic nerve regeneration, there are unknowns, such as the methods to eliminate the risk of anterior segment ischemia and phthisis bulbi. CONCLUSIONS: Globe avulsions with a complete optic nerve cut remain a challenging problem. More research is required to better understand the pathophysiology of optic nerve repair. PMID- 15751251 TI - Congenital dacryocystocele with intranasal extension. AB - PURPOSE: Congenital dacryocystocele is a rare anomaly in the newborn child. The swelling of lachrymal sac is observed by birth and it is associated with obstruction of lachrymal system either above or below lachrymal sac. METHODS: Diagnosis was made by clinical observation. Some ancillary examinations, such as ultrasonography, tomography, and rhinoscopy, were useful. RESULTS: The authors describe the clinical case of a newborn with a unilateral congenital dacryocystocele. This anomaly was successfully treated with probing and marsupialization of the nasal cyst. CONCLUSIONS: Treatment of this congenital anomaly is by light compressive massage, probing with silicone intubation of lachrymal system to assure prolonged permeability of the system, or with marsupialization of the nasal cyst. In some cases with intranasal extension of dacryocystocele, collaboration with an otolaryngologist may be necessary. PMID- 15751252 TI - Subcutaneous tumor of the lower eyelid: a potential manifestation of a Dirofilaria repens infection. AB - PURPOSE: To report a case of Dirofilaria repens presenting as a subcutaneous tumor of the lower eyelid. METHODS: Interventional case report. RESULTS: A 29 year-old man of Greek origin without systemic symptoms presented with a 3-week history of a small painless mass localized in the medial part of the lower eyelid. There was no history of a preceding trauma, injury, or visual impairment; however, the patient had recently been on a holiday in Italy. The lesion persisted after systemic antibiotic treatment. Routine blood tests were normal and the efferent tear ducts were patent. Upon surgical intervention a yellowish, pea-sized cyst-like structure was found beneath the orbicularis muscle and removed in toto. Histologic examination revealed the presence of a wormlike structure with the characteristic features of a single adult Dirofilaria repens nematode. CONCLUSIONS: Infection with the nematode Dirofilaria repens has to be considered in the differential diagnosis of malignant and benign tumors of subcutaneous periocular tissues in patients who traveled to endemic areas. PMID- 15751253 TI - Incomplete extrusion of an acrylic punctum plug in a case of severe dry eye syndrome. AB - PURPOSE: To report a case of extrusion of a new soft punctum plug with thermoexpansion property (Medennium SmartPLUG). METHODS: A soft punctum plug was implanted in a 32-year-old woman with a severe dry eye syndrome in juvenile arthritis. RESULTS: One week after implant the plug partially extruded outside the punctum. Despite this adverse event, all subjective dry eye symptoms improved. CONCLUSIONS: The peculiarity of this case is the persistence of clinical efficacy of the soft punctum plug even if partially extruded. The patient experienced relief of symptoms that can be compared to the benefits usually obtained with a successfully implanted silicon plug. PMID- 15751255 TI - Ocular manifestations in patients with microscopic polyangiitis. AB - PURPOSE: To describe ocular manifestations in patients with microscopic polyangiitis. METHODS: Two patients with microscopic polyangiitis complained of ocular symptoms and underwent ophthalmologic examinations. RESULTS: An 83-year old woman (Case 1) was diagnosed with microscopic polyangiitis, according to the general clinical findings and the presence of perinuclear pattern of antineutrophil cytoplasmic antibodies (P-ANCA). She had hypopyon iridocyclitis in the right eye and retinal cotton-wool spots in the left eye. The patient was treated with oral prednisolone and subconjunctival betamethasone. The hypopyon iridocyclitis and retinal cotton-wool spots responded. A 79-year-old man (Case 2) had bilateral scleritis. The diagnosis of microscopic polyangiitis was made based on general clinical findings and the presence of P-ANCA. Scleritis was reduced after corticosteroid treatment. CONCLUSIONS: Ophthalmologists should be aware that hypopyon iridocyclitis, cotton-wool spot, and scleritis could occur in patients with microscopic polyangiitis. PMID- 15751254 TI - Rhodotorula rubra keratitis and melting after repeated penetrating keratoplasty. AB - PURPOSE: To describe a rare case of Rhodotorula rubra keratitis in a corneal graft, and to discuss the management of this unusual pathogen. METHODS: A 78-year old debilitated man was treated for corneal abscess and descemetocele in his right eye. Urgent penetrating keratoplasty (PKP) with old donor material was performed. The patient underwent a new PKP when appropriate donor cornea was available. The graft was clear after the operation. Corneal cultures were negative. RESULTS: One month after the second PKP procedure, deep stromal infiltrate with hypopyon appeared. Cultures grew R. rubra. A third PKP operation was performed. Systemic and topical amphotericin B treatment was started. Six months after the last operation the graft remains clear. CONCLUSIONS: Rhodotorula keratitis is an extremely rare infection that should be considered in debilitated patients with persistent and progressive corneal infection despite adequate antibiotic therapy. PMID- 15751256 TI - Optical coherence tomography findings in a case of acute multifocal posterior placoid pigment epitheliopathy (AMPPPE). AB - PURPOSE: To report the optical coherence tomography (OCT) findings in a case of acute multifocal posterior placoid pigment epitheliopathy (AMPPPE). DESIGN: Case report. METHODS: A 26-year-old woman was seen for a mild loss of visual acuity in both eyes. The fundus examination revealed yellow-white placoid lesions in the posterior pole suggesting AMPPPE. Fluorescein and indocyanine green (ICG) angiography and optical coherence tomography (OCT) were performed 2 days, 9 days, and 1 month after the first examination. RESULTS: In the acute phases the OCT revealed a mild hyperreflective area above the retinal pigmented epithelium (RPE) in the photoreceptor layer. In the later phases the OCT scan revealed a nodular hyperreflective lesion on the plane of the RPE with mild underlying backscattering. CONCLUSIONS: In AMPPPE, the OCT demonstrates hyperreflective lesions that may indicate inflammatory tissue and inflammatory cells or the presence of ischemic edema in the outer retinal layers. PMID- 15751257 TI - Multiple bilateral choroidal metastatic tumors from a small-cell neuroendocrine carcinoma of unknown primary site. AB - PURPOSE: To report one case of multiple and bilateral choroidal tumors from a poorly differentiated small cell neuroendocrine carcinoma of unknown primary. METHODS: The case of a 30-years-old white female who developed multiple and bilateral choroidal tumors from a poorly differentiated small cell neuroendocrine carcinoma of unknown primary is presented. RESULTS: The patient had a disseminated disease and died 6 months after. The oncologic work-up, including physical examination, laboratory and radiographic study, fails to identify the primary site. CONCLUSIONS: Intraocular involvement from a poorly differentiated small cell neuroendocrine carcinoma of unknown primary has not yet reported. We describe this case together with a review of the literature. PMID- 15751258 TI - Cytomegalovirus retinitis in a patient with non-Hodgkin's lymphoma: a diagnostic dilemma. AB - PURPOSE: Patients with lymphoma can rarely develop cytomegalovirus (CMV) retinitis. Clinically it is difficult to distinguish from intraocular lymphoma. Also, in such cases the CD4+ count may be high. The authors report a rare case of bilateral CMV retinitis in a patient with non-Hodgkin's lymphoma with high CD4+ counts. METHODS: Observational case report with review of literature. RESULTS: CMV retinitis was clinically suspected due to the presence of large areas of retinal necrosis and hemorrhages in one eye and a demarcation line with white mottled retina in the other eye. Other differential diagnoses considered were intraocular lymphomatous infiltration and acute retinal necrosis due to herpes group of viruses. The diagnosis of CMV retinitis was confirmed by polymerase chain reaction performed on vitreous sample. CONCLUSIONS: CMV retinitis can develop in cases of lymphoma despite high CD4+ counts. An early diagnosis can be established by performing PCR on vitreous biopsy. PMID- 15751260 TI - Indocyanine green angiography in congenital hypertrophy of the retinal pigment epithelium. AB - PURPOSE: To report a case of congenital hypertrophy of the retinal pigment epithelium followed up for 7 years showing features not previously reported. METHODS: A complete fundus examination including fluorescein angiography was performed at first examination and at follow-up. RESULTS: The area of congenital hypertrophy of the retinal pigment epithelium presented morphologic changes, showing enlargement of the lesion on one side associated with partial regression on another side. In addition, the indocyanine green angiography findings revealed that this test was more useful than fluorescein angiography to delineate the real boundaries of the lesion and disclosed hypofluorescent areas inside the main lesion that were not appreciated at ophthalmoscopy or fluorescein angiography. CONCLUSIONS: Indocyanine green angiography is an useful test to understand the findings and evolution of congenital hypertrophy of the retinal pigment epithelium. PMID- 15751259 TI - Cystoid macular edema associated with latanoprost after uncomplicated cataract surgery. AB - PURPOSE: To report clinically symptomatic and angiographically documented cystoid macular edema (CME) associated with the use of latanoprost in two pseudophakic eyes after uncomplicated cataract surgery. METHODS: Retrospective review of two patients who had history of latanoprost use and uncomplicated cataract surgery and described blurred vision in the first postoperative month. RESULTS: Ocular examination revealed CME, which was confirmed by fluorescein angiography. The visual acuities of patients improved and the CME was angiographically resolved after discontinuation of latanoprost and the initiation of nonsteroidal anti inflammatory eye-drops and oral acetazolamide. CONCLUSIONS: Until a causal relationship between CME and latanoprost is proved or disproved, caution in its use in pseudophakic patients would be prudent. PMID- 15751261 TI - Optical coherence tomography and fundus microperimetry imaging of spontaneous closure of traumatic macular hole: a case report. AB - PURPOSE: To report a case of spontaneous closure of traumatic macular hole in a young patient followed using optical coherence tomography (OCT) and fundus microperimetry. METHODS/RESULTS: In the right eye of a 10-year-old child, a traumatic macular hole was observed to spontaneously resolve 18 weeks after blunt trauma. Initially, visual acuity in the right eye was 20/200 and OCT examination showed a 200 microm-diameter full-thickness macular hole with perifoveal edema. Fundus microperimetry examination showed an evident decrease in retinal sensitivity within the macular hole and in the upper macular region where an area of commotio retinae was clearly visible. During follow-up OCT demonstrated the appearance of a band of tissue linking the inferior edge of the hole to the foveal retinal pigment epithelium and at the bottom of the hole the presence of hyperreflective (glial) material. Eighteen weeks after trauma right eye visual acuity had improved to 20/25, OCT examination showed a restored foveal depression, and fundus microperimetry demonstrated an increase in foveal sensitivity. CONCLUSIONS: Both OCT and fundus microperimetry were useful tools for following the favorable course in a case of spontaneous closure of traumatic macular hole in a young patient. During follow-up OCT examinations were able to demonstrate the course of macular hole closure. PMID- 15751262 TI - Delayed atrophy of the retinal pigment epithelium after submacular surgery. AB - PURPOSE: We report a case of delayed atrophy of the retinal pigment epithelium (RPE) eighteen months after apparently successful excision of submacular choroidal new vessels (CNV) in a patient with age-related macular degeneration (AMD). METHODS: Case report. RESULTS: Submacular surgery for CNV was achieved without visible disturbance of the underlying RPE in an 83 year old man diagnosed with AMD. At the time of surgery the CNV displayed clinical features consistent with lying internal to Bruch's membrane (Type 2 configuration). There was no visible RPE defect at the fovea and vision improved during the subsequent 12 months follow-up. Eighteen months later, however, an atrophic central RPE defect appeared, with a similar shape to the CNV originally excised. CONCLUSIONS: This case demonstrates that submacular CNV with Type 2 configuration can occur in AMD and lead to an initially favourable outcome following submacular surgery. Atrophy of the RPE nevertheless did eventually occur and in a pattern consistent with damage during the original operation. It is important to consider results of longer term follow-up when interpreting success rates for surgery in AMD. PMID- 15751263 TI - Intraoperative antibiotics and bacterial contamination of the anterior chamber. PMID- 15751264 TI - A clinician's review of the WHI-related literature. AB - When the monitoring board of the Women's Health Initiative (WHI) canceled the estrogen-progestin arm of the study in July 2002, the effect was immediate and dramatic, as several million postmenopausal women with the full agreement of their physicians ceased taking combined hormone therapy. Soon thereafter the manufacturers of conjugated equine estrogens felt compelled to publicize a drastic restriction of the indications for their product. Little notice, except in the medical literature, was given to the continuation of the other treatment arms of the WHI, nor did the rather small (however significant) increases in risk of cardiovascular disease and breast cancer resulting from combined therapy receive widespread serious analysis. In this article, special attention is given to the population sampling involved in setting up the WHI, arm by arm, with full discussion of how these samplings compare with those in other studies--HERS, ERA, WEST, etc. All studies are scrutinized in terms of treatment regimens, follow-up, confounding factors, particularly statins and aspirin, and high drop-out rates in order to discover possible reasons for the results in the WHI for primary and secondary prevention of cardiovascular disease in the combined-therapy arm and slightly disappointing results for breast cancer. Each of the two main sections of the article, Cardiovascular Disease and Breast Cancer, concludes with a detailed summation of points derived from the often contrasting results of the various studies, which can be used in counseling patients. PMID- 15751265 TI - Under-use of emergency contraception for victims of sexual assault. AB - BACKGROUND: Approximately 700,000 women in the reproductive age group are victims of sexual assault in the United States per year. Between 1% and 5% of sexual assaults result in pregnancy, for a total of 32,000 pregnancies per year. Of these, 14,000 are aborted because of incest or rape. OBJECTIVE: To determine the percent of emergency departments in the state of Pennsylvania offering routine counseling and provision of emergency contraception to victims of sexual assault. Secondary objectives were to compare provision practices for Catholic versus non Catholic hospitals, and to compare these practices with other services, such as sexually transmitted disease prophylaxis and sexual assault counseling. METHODS: A 15-item survey instrument was designed to determine the volume of sexual assault patients seen per year, routinely offered services, and emergency contraception protocols. Three telephone callers administered surveys, using a pre-designed script for each call. RESULTS: Of the 165 eligible hospitals, 125 (76%) replied. Less than half (42%) of all hospitals routinely offer emergency contraception counseling, and 16% of the hospitals did not offer any counseling regarding emergency contraception. CONCLUSION: Provision of emergency contraception to victims of sexual assault is inconsistent and insufficient. It is important that sexual assault patients not be further victimized by a system that fails to meet their needs. PMID- 15751266 TI - An economic model to reduce the cost of chemotherapy for gynecologic cancer. AB - OBJECTIVE: Chemotherapy with innovative state-of-the-art medicine at university level can be very costly. Reimbursement until 2002 was at a flat rate, often not even covering the costs of the pharmaceutical substances. To avoid debt a more cost-effective chemotherapy management system had to be found. MATERIALS AND METHODS: From this background, an economic model with four steps was developed: 1. Analysis of current financial situation; 2. Precalculation of chemotherapy costs; 3. Assignment to an individual cost-covering reimbursement pathway; and 4. Postcalculation for cost efficiency and elimination of potential mistakes. RESULTS: After successful implementation of this model we were able to reach cost effectiveness for our chemo unit within 12 months and pay back previous debts and were even able to employ new medical staff. CONCLUSION: With this model we are now able to perform chemotherapy cost effectively at a university level without reducing standard of care. PMID- 15751267 TI - Toremifene, a new agent for treatment of mastalgia: an open study. AB - OBJECTIVE: Endocrine agents have been widely used in the treatment of mastalgia. Toremifene is an agent that predominantly has antiestrogenic properties with minimal estrogenic activities. This study was aimed at investigating this drug in the treatment of mastalgia and to evaluate its tolerability and efficacy. METHODS: Seventeen premenopuasal women with a mean age of 37.7 years complaining of moderate to severe mastalgia received toremifene 60 mg daily. The treatment period was 12 weeks. 70% of patients had cyclical and 30% had noncyclical mastalgia. RESULTS: All women with cyclical mastalgia responded to toremifene compared with only 75% of those with non-cyclical mastalgia. Four patients withdrew from the study after 4 weeks because of side effects, accounting for 23.5% of patients in the study. CONCLUSION: This small study has shown that toremifene is an effective agent in the treatment of mastalgia, but a high incidence of side effects makes it ineligible as an agent of choice for treatment of mastalgia. PMID- 15751268 TI - Involvement of cathepsins in the invasion, metastasis and proliferation of cancer cells. AB - Tumor cell invasion and metastasis are associated with the proteolytic activity of various types of proteinases. Among them, cathepsins, which are lysosomal proteinases, have received more attention recently. Since elevated expressions of cathepsins and diminished levels of their inhibitors have been observed in several human cancers, including breast, gastric and prostate cancer, especially in aggressive cancer cells, cathepsins have been suggested to be biological markers of malignant tumors and have proved useful for prognosis of the disease. Furthermore, cathepsins have various roles in cancer progression. Cathepsin D has a mitogenic activity independent of its proteolytic activity and it attenuates the anti-tumor immune response of decaying chemokines to inhibit the function of dendritic cells. Cathepsins B and L have been shown to play an important role in matrix degradation and cell invasion. The administration of their inhibitors prevents the invasion and metastasis of cancer cells. These results indicate that cancer cells orchestrate various cathepsins to progress malignant diseases. Cathepsins may be a potential target for cancer therapy. PMID- 15751269 TI - Background exposure to PCDDs/PCDFs/PCBs and its potential health effects: a review of epidemiologic studies. AB - Here we review epidemiologic studies dealing with the dietary intake and the body burden of polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzo furans (PCDFs)/ polychlorinated biphenyls (PCBs) in the general population, and potential adverse health effects of these substances, especially on the risk of diabetes mellitus and endometriosis, and on thyroid function and the neurodevelopment of infants. The mean or median intake of dioxin-related compounds among the general populations of various countries is lower than the maximum tolerable daily intake (TDI) set by the WHO in 1998 (4pg TEQ/kg/day). However, there have been few reports on the distribution of intake and the proportion of subjects whose exposure levels exceed the maximum TDL. At present, it remains unclear whether background exposure to dioxin-related compounds is associated with increased risk of diabetes (because of lack of longitudinal studies), endometriosis (because of lack of studies with sufficient statistical power), or altered thyroid function (because of inconsistent results on humans). Consistent results have been reported for the association between exposure to background levels of PCBs/dioxins, especially trans-placental PCBs, and defective neurodevelopment of infants in the U.S. and Europe. Thus, efforts should be made to further decrease the body burden among women of reproductive age by reducing the release of PCDDs/PCDFs/PCBs into the environment. PMID- 15751270 TI - Living-donor liver transplantation: present status and future perspective. AB - About 15 years have passed since the first liver transplant from a living donor (living donor liver transplantation: LDLT), and the status of the procedure has since been established as a standard cure for end-stage liver disease in Japan where liver transplantation (LTx) from deceased donors has not yet been accepted. However, the following problems are surfacing with the increase in the number of LDLTs between adults: graft size mismatching, an ABO blood-type incompatible transplantation, the expansion of LDLT indication to hepatocellular carcinoma (HCC), the relapse of hepatitis C after LDLT, marginal donors, and the freedom from immunosuppressive treatment. In this article we outline the present conditions of these problems and the future view of the LDLT. PMID- 15751271 TI - Overweight is associated with allergy in school children of Taiwan and Vietnam but not Japan. AB - We collected information concerning diagnosed allergy from 2027 school children in Japan, Taiwan and Vietnam. Children were classified according to the age and sex-specific body mass index (BMI) per-age as indicator of weight status. Logistic regression was performed to examine the relationship between percentiles of BMI-per-age and allergy. Compared with children at the lowest percentile group Taiwanese children at > 85th percentile group showed a tendency toward higher risk of allergy (OR = 1.79, 95% CI 0.98 to 3.27; p = 0.060). When children with rhino-conjunctivitis were excluded from the analysis the association reached statistical significance (OR = 2.89, 95% CI 1.08 to 7.75; p = 0.035). Vietnamese children at > 85th percentile group showed a significantly higher risk of allergy (OR = 2.34, 95% CI 1.06 to 5.17; p = 0.035). This association was not observed when children with atopic dermatitis or food allergy were excluded from the analysis, although a tendency toward increased risk of allergy at BMI-per-age > 85th percentile remained. Our study sample of Japanese school children showed no association between being overweight and allergy. PMID- 15751273 TI - Findings of brain 99mTc-ECD SPECT in high-functioning autism--3-dimensional stereotactic ROI template analysis of brain SPECT. AB - The aim of this study is confirmation of an abnormal regional cerebral blood flow (rCBF) pattern in high-functioning autism (HFA). Confirmation of an abnormal rCBF pattern in HFA may be useful for elucidate of its pathophysiology and a differential diagnosis, such as with attention-deficit/hyperactivity disorder (AD/HD). Brain 99mTc-ECD SPECT was performed in 16 cases of HFA. The HFA group consisted of 16 cases of HFA. They were all male, with an IQ of 76-126. They had normal brain MRI findings, and had an age of 9-14 years. We examined abnormal rCBF in HFA by comparing the results to those in the control group. The control group consisted of 1 male and 4 females cryptogenic epilepsy patients with normal intelligence. They have no problems in learning at school or mental or behavioral traits. They had normal brain MRI or SPECT findings, and had an age of 7-15 years. 3-dimensional stereotactic ROI template (3DSRT) was used to analyze SPECT data. We calculated the 'relative rCBF (%)' (RI count of each segment x 100/Sum of RI count of the corresponding hemisphere), and compared the values between the two groups. We found a significantly low 'relative rCBF (%)' in the left temporal region in the HFA group. We also calculated the 'L/R ratio' (the 'relative rCBF (%)' of a segment on the left side / the 'relative rCBF (%)' of the corresponding segment on the right side), and compared the value for each segment between the two groups. There were no significant differences in any segments between the two groups. We also checked for differences in the 'relative rCBF (%)' between segments on the right side and corresponding segments on the left side in both the HFA and control groups. We found significant right < left perfusion in the angular region and significant left < right perfusion in the pericallosal, thalamus, and hippocampus region in the HFA group. We also found significant right < left perfusion in the temporal region in the control group. Significant hypoperfusion in the left temporal region due to an unidentified underlying brain pathology and abnormal laterality in the angular, temporal (lack of right < left perfusion), pericallosal, thalamus, and hippocampus regions may influence the symptoms of autism. PMID- 15751272 TI - Resistance to topoisomerase II inhibitors in human glioma cell lines overexpressing multidrug resistant associated protein (MRP) 2. AB - For understanding of the resistance to topoisomerase II inhibitors, 50 sublines were isolated as single clones from parental glioma cell lines by exposure to VP 16 or m-AMSA. The quantitative aspects of topoisomerase II alpha,multi drug resistant gene (MDR)-1, breast cancer resistance protein (BCRP), and multidrug resistant associated protein (MRP) 1-5 were studied by Northern blotting in 50 resistant cell lines. By understanding the function of MRP2, we picked up three drug resistant sublines (T98G-ml, T98G-m2, and gli36-VP1) that overexpressed MRP2, but did not overexpress MDR-1 or MRP1-5 except 2. Moreover, in the results of northern blot analysis of mRNA for topoisomerase II alpha identical results are observed in parental cell lines and their resistant cell lines, suggesting that alterations in topoisomerase II do not account for the resistance in these cells. To determine whether the cellular sensitivity to anticancer agents was closely associated with the cellular levels of MRP2, we established cell lines with the same levels of MRP2 as their parental cells by introducing the MRP2 antisense expression plasmid into resistant cells. Etoposide (VP-16) accumulation and efflux studies were carried out in the parental cell lines and their drug resistant cell lines. Decreases in the HS-VP-16 accumulation and increases in the efflux were observed in these drug resistant cell lines. In the cytotoxicity assay, these drug resistant cell lines were resistant to multiple topoisomerase II inhibitors with little cross resistance to vincristine, and display efflux of VP-16. We found that the resistant cells transfected with MRP2 antisense cDNA displayed increased cellular levels of VP-16 and enhanced sensitivities to topoisomerase II inhibitors. In this study on the T98G-ml, T98G-m2, and gli36-VP1 cell lines, we showed a high correlation between MRP2 mRNA and VP-16 efflux, suggesting that MRP2 could be a new transporter for topoisomerase II inhibitors. PMID- 15751274 TI - No association of the Trp 64 Arg mutation of the beta3-adrenergic receptor gene with obesity, type 2 diabetes mellitus, hyperlipidemia, and hypertension in Japanese patients with schizophrenia. AB - OBJECTIVE: This study was conducted to address the question of whether the beta3 adrenergic receptor gene mutation (Trp 64 Arg) is associated with metabolic disease in Japanese patients with schizophrenia. METHODS: In a cross-sectional study, 89 participants were grouped into three genotypes. The 64 Arg allelic frequency in patients with or without metabolic disease was analyzed. Anthropometrics variables and biochemical parameters were compared among the genotypes. RESULTS: The 64Arg allele, which had a frequency of 0.22, was not associated with obesity, type 2 diabetes mellitus, dyslipidemias, or hypertension. No significant differences among the genotypes were found in current age, age at diagnosis with schizophrenia, body mass index, waist-hip ratio, plasma glucose, plasma insulin, triglycerides, free fatty acids. Patients with the 64 Arg allele had greater 24-h excretion of norepinephrine than those lacking the variant (p = 0.019). CONCLUSION: The 64 Arg allelic mutation is not associated with obesity, type 2 diabetes mellitus, lipid metabolism dysfunction, or hypertension in Japanese patients with schizophrenia. PMID- 15751275 TI - Inhibitory effects of asiatic acid and CPT-11 on growth of HT-29 cells. AB - Asiatic acid is a pentacyclic triterpene contained in medicinal plants. The cytotoxic effect of this compound and its augmentative effect on the anticancer drug irinotecan hydrochloride (CPT-11) were investigated in the human colon adenocarcinoma cell line HT-29. Asiatic acid dose-dependently showed cytotoxicity in HT-29 cells. DNA fragmentation, annexin-positive apoptotic cells, and caspase 3 activation were observed in a dose-dependent manner. A caspase-3 inhibitor suppressed the DNA ladder formation in a concentration-dependent manner. Bcl-2 and Bcl-XL proteins were decreased by asiatic acid treatment. These results indicate that asiatic acid induced apoptosis in HT-29 cells via caspase-3 activation. Cytotoxic effects of combined treatment with CPT-11 and asiatic acid on HT-29 cells were further examined. Simultaneous treatment or sequential exposure first to asiatic acid and then to CPT-11 showed an additive effect. Synergism was observed when cells were first exposed to CPT-11 and then to asiatic acid. These results suggest that asiatic acid can be used as an agent for increasing sensitivity of colon cancer cells to treatment with CPT-11 or as an agent for reducing adverse effects of CPT-11. PMID- 15751276 TI - Lower axillary dissection in breast cancer surgery may be candidate for cases with early breast cancer. AB - Lower axillary lymph node dissection (lower parts of both the level I and II elements below the second intracostobrachial nerve) and level I and II lymph node dissection were performed on breast cancer patients (n = 54), and the results with the two methods were compared in terms of the status of detected lymph node metastases. For Stage I, N0 cases, the results for pathological classification lymph node metastases (pN) were in agreement between the two dissection methods. And, the occurrence of operated arm swelling wasn't recognized when a side effect was examined with the case (n = 28) that only lower axillary dissection was carried out in case of an operation for breast cancer. Accordingly, it was surmised that lower axillary dissection provides accurate pN information for Stage I, N0 cases. These results indicate that lower axillary dissection has the potential to become an effective, standard surgical procedure for breast cancer patients whose preoperative disease stage is Stage I. PMID- 15751277 TI - The Totton-Ken Seibu earthquake and exacerbation of asthma in adults. AB - The aim of the study was to characterize patients at risk for exacerbations of their asthma as a result of the Tottori-Ken Seibu earthquake and to identify factors that predict exacerbation of asthma after an earthquake. A retrospective cohort study-analysis was conducted of 156 asthmatic patients, aged 18 to 89 years, who were out-patients of Tottori University Hospital and who had completely recorded their asthmatic symptoms and measured their peak expiratory flow (PEF) rates for more than one year prior to the earthquake. Seventeen (11%) patients who experienced the earthquake were identified as having an exacerbation within one month after the earthquake. Diurnal variability of PEF during the month after the earthquake was compared to values during a matched month one year previously. When factors associated with exacerbation were identified by a review of the medical case notes and the contribution of these factors to the exacerbation was determined using multivariate analysis, airflow limitation was shown to be independently associated with exacerbation after the earthquake. Acute asthma attacks are more likely to occur within the first week after the earthquake event without diurnal PEF variability. Asthma is likely to worsen after an earthquake. PMID- 15751278 TI - Elevation of macrophage-derived chemokine in eosinophilic pneumonia: a role of alveolar macrophages. AB - Macrophage-derived chemokine (MDC/CCL22) and thymus-and activation-regulated chemokine (TARC/CCL17) are ligands for CC chemokine receptor 4. Recently, TARC has been reported to play a role in the pathogenesis of idiopathic eosinophilic pneumonia (IEP). The purpose of this study was to evaluate the role of MDC in IEP and other interstitial lung diseases (ILDs). MDC and TARC in the bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay in patients with ILDs and healthy volunteers (HV). We also examined the expression of MDC mRNA in alveolar macrophages (AM) by real-time quantitative reverse transcriptase-polymerase chain reaction. Both MDC and TARC were detected only in BALF obtained from IEP patients. The concentration of MDC was higher than that of TARC in all cases. The level of MDC in IEP correlated with that of TARC. AM from IEP patients expressed a significantly higher amount of MDC than that from HV at the levels of protein and mRNA. MDC in BALF from IEP dramatically decreased when patients achieved remission. These findings suggest that MDC, in addition to TARC, might be involved in the pathogenesis of IEP, and AM play a role in the elevation of MDC in IEP. PMID- 15751279 TI - The significance of cathepsins, thrombin and aminopeptidase in diffuse interstitial lung diseases. AB - To determine the significance of proteases in interstitial lung diseases, we examined the activity of cathepsins, thrombin, and aminopeptidase in bronchoalveolar lavage (BAL) fluid from patients with these disorders. Significantly increased activities of cathepsin H and aminopeptidase were detected in BAL fluid from patients with idiopathic pulmonary fibrosis (IPF), cryptogenic organizing pneumonia (COP), chronic eosinophilic pneumonia (CEP) and hypersensitivity pneumonitis (HP). Significantly higher activity of cathepsin B was found in BAL fluid from patients with CEP. The activity of thrombin was significantly higher in patients with IPF and CEP. In patients with IPF, there were significant correlations between neutrophil number and the activity of cathepsin B, cathepsin H or aminopeptidase. In patients with COP and HP, the activity of the proteases was significantly higher in patients with higher number of lymphocytes than in those with lower number of lymphocytes. The present study suggests that the activity of the proteases is a useful marker in activity of the interstitial lung diseases, and may have a role in the pathogenesis of these disorders. PMID- 15751280 TI - Simultaneous detection of Bacteroides fragilis group species by leuB-directed PCR. AB - Bacteroides species, saccharolytic Gram-negative obligate anaerobes, are frequently isolated from human infections such as peritonitis, abscesses and bacteremia. Among the species in the genus Bacteroides, the species called "B. fragilis group" are particularly involved in human infections and are medically important because they account for a major part of anaerobic isolates from clinical specimens. The purpose of this study was to develop PCR primers that specifically and simultaneously amplify the beta -isopropylmalate dehydrogenase gene leuB in B. fragilis group species. We determined partial nucleotide sequences of leuB genes and compared them in seventeen strains of nine B. fragilis group species, and the regions that are conserved among Bacteroides strains but different from other species were used as a B. fragilis group specific PCR primer set, BacLBF-BacLBR. Specificity tests of the primer set using 52 phenotypically characterized strains and 75 isolates from rat feces showed only one case each of false-positive and false-negative. The detection limit of the leuB-directed PCR using BacLBF and BacLBR was 3.9 x 10(3) colony-forming units. These results indicate that leuB amplification using BacLBF andBacLBR is a useful tool for rapid diagnosis of Bacteriodes infection and for rapid differential diagnosis of anaerobic infections. PMID- 15751281 TI - Fine mapping of the hyperglycemic and obesity QTL by congenic strains suggests multiple loci on rat chromosome 14. AB - Linkage analysis previously identified a hyperglycemic quantitative trait locus (QTL), Nidd 2/of, on rat Chromosome 14 in crosses utilizing OLETF (Otsuka Long Evans Tokushima Fatty) rat, a model for type 2 diabetes. A separate QTL study mapped an obesity QTL, Obs5, to the same chromosomal region. A congenic strain placing ca. 38 cM OLETF-derived segments containing both Nidd2/of and Obs5 on the F344 background was shown to possess mild diabetic and obese phenotypes, suggesting the presence of mutations affecting the glucose metabolism and fat accumulation. In order to localize the loci more precisely, we generated a series of deletion-subcongenic strains in which OLETF-segments were shortened from either ends. We found that there are at least two hyperglycemic QTLs within the Nidd2/of locus. We predict that they are localized towards both ends of the Nidd2/of region. In contrast, Obs5 QTL was further narrowed down to a single region of ca. 10 cM fragment. PMID- 15751282 TI - Two cases of retroperitoneal hematoma caused by combination of anticoagulant therapy and 5-fluorouracil. AB - We reported two cases of retroperitoneal hematoma in patients who received a combination of anticoagulant therapy and 5-fluorouracil (5-FU). We should be aware of the possible interaction of this combination therapy and monitor prothrombin time (PT) prolongation. CT is useful for evaluation of the disease. PMID- 15751283 TI - A case of sarcoidosis accompanying squamous cell carcinoma in the mandibular gingiva. AB - A 51-year-old man with a history of gingival cancer two years previously was referred to our hospital for further examination of chest abnormal shadow. Bilateral hilar and mediastinal lymphadenopathy, diffuse small nodular opacities and pleural nodules were observed in chest high resolution CT. Serum angiotensin converting enzyme and lysozyme were elevated. Transbronchial lung biopsy specimens demonstrated non-caseous granuloma. CD4-positive lymphocytes were increased in broncho-alveolar lavage (CD4/CD8 ratio 5.47). The patient was diagnosed as having sarcoidosis. Radiological findings were improved and serum angiotensin converting enzyme level was decreased to within the normal range by corticosteroid therapy (prednisolone 30 mg/day). This is the first report of sarcoidosis accompanying the gingival cancer. PMID- 15751284 TI - A patient with octopus pot-shaped cardial cancer that metastasized to multiple organs. AB - A 71-year-old male was admitted for abdominal fullness. The condition rapidly deteriorated in a short period (3 weeks), and the patient died. Autopsy revealed a protruding lesion measuring about 3 cm with erosion measuring 5 mm in diameter immediately below the esophago-gastric conjugation site, suggesting primary cardial undifferentiated adenocarcinoma. In the primary focus, changes on the mucosal surface were almost normal However, below the mucosa, infiltration of cancer cells was observed in an approximately 3 cm area along the gastric wall. Simultaneously, the site of infiltration was markedly increased in deep areas. Extraserous infiltration was observed. The morphology was special, and resembled an octopus pot, a trap used to catch octopuses in Japan, with a narrow top and a broad base. In our patient, metastatic lesions were detected in multiple organs, including the stomach. PMID- 15751285 TI - [The sanitary and epidemiological situation in the Baikal Region and its influence on human health]. PMID- 15751286 TI - [Influence of environmental factors on human health (in case of Orekhovo Zuyevo)]. AB - The paper analyzes the actual nutrition in 301 schoolchildren (142 children and 159 adolescents), the state of ambient air and the health status of the population in 5 districts of the town of Orekhovo-Zuyevo. Non-balanced diets and ambient air pollution have been ascertained to negatively affect human health. PMID- 15751287 TI - [Soil heavy metal pollution in Ryazan: ecological and hygienic aspects]. AB - The paper presents the results of investigations evaluating the soil pollution with heavy metals in Ryazan. The authors have determined the priority pollutants, geochemical anomalies on the territory of the town and the qualitative and qualitative composition of the pollutants in relation to the source of waste and to the specific features of an atmospheric and technogenic transfer. They propose measures to improve the urban ecological and sanitary status, which are associated with the utilization of the plants that accumulate heavy metals and with the replacement of the polluted soil with artificial soil-like masses. PMID- 15751288 TI - [Risk factors of the intra-housing environment to human health]. PMID- 15751289 TI - [Seasonal features of the formation of ambient air pollution in Almaty]. PMID- 15751290 TI - [Breast cancer in Chapayevsk]. AB - Numerous epidemiological surveys show the ambiguity of evaluation of stable organic chlorine compounds as a risk factor for breast cancer (BC). This is likely to be, to a certain degree, associated with the specific features of performance of epidemiological surveys. The selection of cases of BC from different areas has provided groups with different levels of exposure that as judged from the content of the above substances in the biological environments. Only few epidemiological surveys have been made in the world to assess stable organic pollutants (SOP) as risk factors of BC mainly in occupational groups. In the local areas highly polluted with dioxins, there is evidence for their role as a risk factor of BS only for women living in Sevezo. These findings are in agreement with the results of the surveys made in Chapayevsk. The analytical epidemiological study using a case-control approach has revealed the significance of biological (BC in relatives, childlessness), occupational (women's work at CVZH) and environmental (pollution, the more frequent ingestion of local pork and fish than in the control group) risk factors of BC. In the 1950-1960s, the largest Russian enterprises of chlorine chemistry worked most actively not only in Chapayevsk, but also in towns with plants of chlorine industry, such as Dzerzhinsk, Novomoskovsk, Sayansk, Usolye-Sibirskoye, and others. Ecological and epidemiological studies should be organized in these towns in order to define the value of SOP as a risk factor of BC for general populational groups. PMID- 15751291 TI - [Hygienic aspects of a human health-hot water supply quality warm relationship]. PMID- 15751292 TI - [Impact of working conditions on morbidity accompanied by temporary disability in workers at the present-day tobacco plants]. AB - By analyzing morbidity accompanied by temporary disability in workers at two tobacco plants, the authors conclude that their working conditions and the organization of medical aid to them have an impact on the pattern of morbidity and the health status of the workers. PMID- 15751293 TI - [Some aspects of professional activities and a risk to the health status of aircraft designers]. AB - Morbidity associated with temporary disability and risks for major syndromes have been studied in aircraft designers who work under the conditions regarded as hazardous (the third degree of Class 2). The risks for major syndromes have been analyzed in relation to sociopsychological characteristics. The manifestations of the emotional exhaustion syndrome have been examined in the workers exposed to industrial stresses. PMID- 15751294 TI - [Psychophysiological professional selection of drilling technicians who provide the sinking of oil wells]. AB - To protect health and to warn future workers against the choice of a petroleum job that was not in accord with their psychological and physiological features of their organism, studies (such as sanitary and psychophysiological ones) were conducted to create criteria for the professional selection of drilling technicians who provided the sinking of oil wells. The studies have been systematized and mathematically analyzed. This made it possible to reveal the significant psychophysiological properties (8 parameters) in a drilling technician. By taking into account these properties, the author derived a linear equation that estimates the fitness of a candidate for a job as a drilling technician. PMID- 15751295 TI - [Iodine deficiency as a factor that diminishes the mental and intellectual development of a child]. AB - Iodine deficiency in child is one of the factors that diminish his/her mental and intellectual development. The impact of the severity of iodine deficiency on both intellect and memory has been studied. It has been established that reproduction of verbal information diminishes and visual memory and other mental activity deteriorate in iodine deficiency. The number of accentuated personalities increases in the groups of children with decreased iodine levels. Self overestimation occurs more frequently in these groups than that in a group of children with normal levels of this trace element. The manifestation of egocentrism and aggression increases in proportion with the more severity of iodine deficiency. PMID- 15751297 TI - [Central hemodynamic parameters in adolescents aged 10-17 years in the Primorye Territory]. AB - The paper presents the results of their own studies of the central hemodynamics of 1900 adolescents aged 10-17 years in the Primorye Territory. Most adolescents were found to have reduced cardiac output, stroke output, and cardiac index and no definitive level of the development of the cardiovascular system by the age of 17 years; the adolescents aged 10-14 years had diminished physical fitness. ECG arrhythmia was most frequently observed in the schoolchildren. There is no question that to study the adolescents' morphofunctional development, by applying a regional approach is important and necessary for the substantiation and implementation of preventive measures. PMID- 15751296 TI - [Hygienic training of students, the future teachers of primary forms]. AB - The paper emphasizes the need for improving the hygienic training of pedagogical students under the conditions of the present-day developing education. It proposes a stepwise system of hygienic training through the subjects of federal and national-regional components, special courses, and pedagogical practice. PMID- 15751298 TI - [The physiometric parameters in children living in the areas having different levels of aerotechnogenic load]. AB - The physiometric parameters were studied in 1006 children living in the areas having different level of ambient air pollution. It was found that the basic physiometric parameters were decreased in children living in more unfavorable environmental areas. PMID- 15751299 TI - [Impact of physical exercises on the cardiovascular and respiratory systems of students]. AB - The paper deals with the impact of physical classes at a higher educational establishment on the cardiovascular and respiratory systems of students. First- and second-year students who did physical exercises in accordance with the curriculum and fourth-year students who did not were followed up. Blood pressure and heart rate before and after exercise and the maximum oxygen consumption were used as parameters that characterized the state of the cardiovascular and respiratory systems. No physical exercises in third- and fourth-year students were shown to negatively affect their cardiorespiratory system. The current introduction of physical exercises at all courses of higher educational establishments is positively appreciated. PMID- 15751300 TI - [Analysis of a school timetable, bearing in mind the health status of children]. AB - A type of medical examination of a school timetable is proposed, by bearing in mind the actual educational price established at a specific educational establishment. The ratio of the physiological and emotional stress in pupils during schooling to their learning progress and physical fitness is the price of schooling. The study has revealed no direct relationship between the level of schooling load and the price of education. There are difficulties in learning within the first days of a week and a need for additional efforts to maintain the pupils' working capacity at the end of a week. PMID- 15751301 TI - [Impact of the mineral composition of drinking water on the health status of children]. AB - The study was undertaken to evaluate the effect of highly mineralized drinking water on the health status of children. Two Kazan districts that greatly differed in the conditions of water supply and in the mineral composition of water were chosen to define a relationship of the health indices in children to the chemical composition of water. Eight hundred and thirty three schoolchildren aged 7-9 years were interviewed by a questionnaire and examined for their objective status. The urinary levels of trace elements, such as copper, zinc, cadmium, and gross elements, such as calcium, magnesium, were measured by atomic absorption spectrophotometry on an AAS-SA 10 MP device. PMID- 15751302 TI - [Methodological approaches to evaluating the adaptation of schoolchildren under the conditions of iodine deficiency and ambient air pollution]. PMID- 15751303 TI - [Prospects of sociohygienic monitoring in the nosological and general pathological contexts of medicine]. PMID- 15751304 TI - [The systems principle in establishing the allowable human daily dose of pesticides]. AB - Instead of the earlier isolated standardization of the levels of pesticides in the foodstuffs, water, air, a comprehensive hygienic regulation of pesticides in the environmental objects, which is based on the criterion--an allowable daily dose (ADD)--has been implemented since 1967. The systems principle is used, by taking into account the toxicological and hygienic parameters and the adverse reactions. Tens of thousands of regulations have been legally adopted. ADD has been shown to be the basis for constructing a comprehensive system of regulation of pesticides, including the "expectation time", "the time of going to work", etc. The principles of harmonization of ADDs adopted by the WHO and Ukraine are presented. PMID- 15751305 TI - [Principles in the standardization of chemical compounds having an olfactive effect in the working area air]. PMID- 15751306 TI - [Choice of safety criteria during accidents occurring at the dangerous industrial objects during work using cadmium and metallic mercury]. AB - Based on the systematization of the experimental and epidemiological data on the toxicity of the compounds of mercury and cadmium and on the concept of a reasonable risk, the author has developed qualitative indices (criteria) required to analyze the safety of work associated with metallic mercury and cadmium at the dangerous industrial objects while declaring their safety. The fulfillment of these criteria ensures the high level of safety of mercury- and cadmium associated work; it is in accord with the current trends in approaches to evaluating the safety of devices using toxically dangerous substances. The criteria considers the risk of human death due to the simultaneous intake of mercury vapors and cadmium oxide aerosols during accidents, as well as the level of chronic intake of these substances in different groups of persons on secondary dust formation (evaporation) after an accident. PMID- 15751307 TI - [Hygienic standardization of the levels of selenium hexafluoride in the air of a working area]. AB - The paper presents the results of investigations on the hygienic standardization of the levels of selenium hexafluoride (SeF6) in the air of a working area. It also analyzes the data on the effects of SeF6 inhalation on laboratory animals, by applying morphological, physiological, hematological, and biochemical studies. The studies have yielded the basic parameters of inhalation toxicity and established the threshold of the irritant action of the substance, developed and approved its maximally acceptable concentration (0.2 mg/m3, gas + vapors) and a procedure for photometric determination of SeF6 in the air of a working area. PMID- 15751308 TI - [Informational and educational work on the prevention of respiratory diseases in the Krasnoyarsk Territory]. PMID- 15751309 TI - [Analysis of the situation established in connection with the altered drinking water quality in Izhevsk, Udmurt Republic, in 2003]. PMID- 15751310 TI - [An open-drop crystalloscopic procedure for studying serum in the toxicological and-hygienic experiment]. PMID- 15751311 TI - [A procedure for determination of the content of sodium polyacrylate in aqueous extracts]. PMID- 15751312 TI - [An experience of monitoring the quality of laboratory filters for sanitary microbiological examination of water]. PMID- 15751313 TI - [Biological evaluation of water quality: ways of improvement]. PMID- 15751314 TI - [Aleksei Alekseevich Minkh (on the occasion of his 100th birthday)]. PMID- 15751315 TI - [Energy losses from a short bowel: not only fat]. AB - Two adult patients, men aged 43 and 45 years, with Crohn's disease, complicated by high-output small-bowel-stomy and short-bowel syndrome due to several intestinal resections, presented with extreme weight loss. Although both patients followed a high-calorie diet combining solid foods and enteral nutrition by nasogastric tube, containing 16,000 kJ and 21,000 kJ, respectively, weight loss continued. Faecal fat excretion and basal metabolic rate were determined, but these could not explain the caloric deficit. Therefore, faecal bomb calorimetry, a measurement of total faecal energy content, was also performed, revealing a considerably higher faecal energy loss than had been calculated from faecal fat excretion; this indicates that faecal carbohydrate loss plays an important supplementary role. A stable weightwas achieved in both patients by prescribing extra food. Therefore, faecal fat excretion is an insufficient indicator of total faecal calorie loss in patients with high-output stomata and short-bowel syndrome. Bomb calorimetry may be considered as a tool to determine the remaining absorptive capacity in short-bowel patients. PMID- 15751316 TI - [Treatment of intestinal failure by total parenteral nutrition at home in children and adults]. AB - Intestinal failure is characterised by inability of the intestine to absorb sufficient nutrients to maintain the integrity and function of the body. This can be caused by malabsorption due to too short an intestine or an abnormality of the mucosa, or by a severe motility disorder. In addition to dietary measures, the prescription of total parental nutrition (TPN) at home is sometimes necessary. This treatment is a burden on the patient and the risk of complications must be reduced to a minimum. The risks of long-term parenteral nutrition can be limited and the quality of the provision of services can be increased if the co ordination is in the hands of a centre for home parenteral nutrition. In the Netherlands there are two centres for home-TPN: the St Radboud University Medical Centre in Nijmegen and the University Medical Centre (AMC) in Amsterdam. In both children and adults, the most common indications are the short bowel syndrome and motility disorders. However, the syndromes that cause this are clearly different in the different age groups. Parenteral nutrition can be given for long periods of time. A large variety of complications can occur, related especially to the equipment or the nutrients. When the nutrition is given via a central venous catheter, then sepsis is a serious and possibly life-threatening complication. In case of administration via an arteriovenous shunt, thrombosis of the shunt is the most frequent problem. If the treatment by means of home-TPN fails, then transplantation of the small intestine should be considered. PMID- 15751318 TI - [Diagnostic image (228). An African infant boy with genua vara]. AB - In an African infant boy with genua vara rickets was diagnosed, and successfully treated with vitamin D and calcium. PMID- 15751317 TI - [Small bowel transplantation as a treatment option for intestinal failure in children and adults]. AB - Small bowel transplantation for intestinal failure is no longer an experimental procedure, but an accepted treatment for patients where total parenteral nutrition (TPN) therapy for intestinal failure is unsuccessful. Early referral for screening for small bowel transplantation should be considered in patients with permanent intestinal failure who have occlusion of more than 2 major veins, frequent line-related septic episodes, impairment of liver function or an unacceptable quality of life. With the increased experience in post-transplant patient care and newer forms of induction (thymoglobulin, IL-2 receptor antagonists) and maintenance (tacrolimus) therapies, the 1-year graft survival has increased to 65% for isolated and to 59% for liver/small bowel transplantation and is further improving. Rejection, bacterial, fungal and viral (Cytomegalovirus, Epstein-Barr-virus) infections, post-transplant lymphoproliferative disease and graft versus host disease are the most common complications after intestinal transplantation. Although most of the long-term survivors are TPN-independent and have a good quality of life, the risk of the procedure and long-term adverse effects ofimmunosuppressive medication limits small bowel, or liver/small bowel transplantation only to patients with severe complications of TPN therapy. PMID- 15751319 TI - [Adolescents with acute lymphatic leukaemia achieve significantly better results when treated following Dutch paediatric oncology protocols than with adult protocols]. AB - OBJECTIVE: To compare the results, in adolescents with acute lymphatic leukaemia (ALL), of treatment according to the protocols of the Netherlands Foundation for Paediatric Oncology (DCOG) or according to the protocols for adults of the Dutch Foundation for Adult Haemato-Oncology (HOVON). DESIGN: Retrospective. METHOD: During the period from May 1985 to November 1999, 120 15-20-year-old adolescents with ALL were treated: 47 according to a DCOG protocol and 73 according to a HOVON protocol. The records of the integrated cancer centres indicate that this represented about two-thirds of all known adolescents with ALL in the Netherlands. RESULTS: Adolescents with ALL treated according to the DCOG protocols had significantly better 5-year survival rates (79% versus 38%), a significantly lower probability of relapse (27% versus 55%) and a lower treatment related mortality (4% versus 25%) than the adolescents treated according to the HOVON protocols. CONCLUSION: This difference in outcome was most likely related to differences in structure and content between the DCOG and HOVON protocols. The HOVON protocols consisted of relatively short, intensive chemotherapy, often followed by autologous or allogeneic bone-marrow transplantation. Several treatment elements present in the DCOG protocols, such as high-dose methotrexate, reinduction therapy and maintenance therapy were absent in the HOVON protocols. Moreover, the cumulative dosages of dexamethasone, mercaptopurine, asparaginase and vincristine were lower in the HOVON protocols. PMID- 15751320 TI - [Fertility after treatment for ectopic pregnancy: evaluation of the switch from laparotomy to laparascopy]. AB - OBJECTIVE: Evaluation of the switch in surgical treatment for ectopic pregnancy from laparotomy to laparoscopy at the Sophia Hospital, Zwolle, the Netherlands. DESIGN: Retrospective. METHOD: Initially an ectopic pregnancy was treated by means of laparotomy at the Sophia Hospital, Zwolle, the Netherlands. At the end of 1996 laparoscopic treatment became the method of first choice. In 2003, all patients who received surgical treatment for ectopic pregnancy in the period 1995 2001 were evaluated. As well as examining their medical records, all patients were questioned about the fertility outcome since the ectopic pregnancy. All of the patients could be traced and the response was 100%. RESULTS: In the study period 97 patients were operated on due to an ectopic pregnancy. The admission period was significantly shorter after laparoscopy compared to laparotomy. The complication rate in both groups was low. In total 78 (80%) of the patients still wanted to conceive. During the observation period, 94 pregnancies occurred in 61 patients. The pregnancy rates following laparotomy and laparoscopy were the same. After salpingostomy, the future fertility was significantly higher (p < 0.05) compared to salpingectomy. CONCLUSIONS: A laparoscopic treatment of ectopic pregnancy is more advantageous for the patient compared to laparotomy: in our results this did not compromise future fertility. Salpingostomy was associated with an increased chance of later pregnancy compared to salpingectomy. PMID- 15751321 TI - [Intestinal rehabilitation for children with short bowel syndrome]. AB - Three children, 2 boys aged 5 and 4 years, and 1 girl aged 2 years, who suffered from intestinal failure as a result of a short bowel syndrome and who were dependent on total parenteral nutrition (TPN), were screened for small bowel transplantation. The girl also had progressive liver failure. Each child had a clear indication for either isolated small bowel or combined small bowel/liver transplantation. All three children were enrolled in an intestinal rehabilitation program because they were referred early. The intestinal rehabilitation consisted of a systematic analysis of the absorptive capacity of the residual intestine, optimizing enteral and parenteral feeding, prevention of catheter sepsis and further deterioration of liver function. The rehabilitation was performed in an in-patient setting. All three children were partially weaned from TPN, while their growth improved. None of the children were consequently listed for small bowel transplantation. When a child is initially referred for small bowel transplantation, intestinal rehabilitation should be considered as part of an intestinal transplantation program. PMID- 15751322 TI - [Defecation disorders in children: treatment with colonic irrigation through an appendicostomy]. AB - In 3 patients with serious constipation or faecal incontinence the appendix was sewn open-ended into the abdominal wall (Malone-stoma) allowing antegrade bowel washouts. The patients were a 4-year-old boy with Hirschsprung's disease, a 15 year-old boy with chronic functional constipation and encopresis, and a 5.5-year old boy with incontinence associated with a meningomyelocele. Following the procedure they were free from bowel problems. Faecal incontinence has a major detrimental effect on the psychosocial development of children. Severe constipation and faecal incontinence which do not optimally respond to conventional therapy can be treated by this procedure. Treatment is effective in around three-quarters of these children. Complications occur in an equal number of children, stenosis being the most frequently seen. PMID- 15751323 TI - [Intra uterine insemination with processed sperm for HIV serodiscordant couples in whom the man is HIV positive]. AB - The desire to have children is more and more common in HIV serodiscordant couples. The Academic Medical Centre Amsterdam has developed a new treatment protocol for couples in whom the man is HIV positive. Semen is processed to obtain HIV-1 free spermatozoa. These spermatozoa are used for IUI treatment. Thus far, 20 serodiscordant couples underwent 76 IUI cycles. An insemination was performed in 50 cycles (66%). The insemination was cancelled in 26 cycles, because of too many follicles (risk for multiple pregnancy), weekend (no possibilities for virological testing), not enough spermatazoa after preparation, a positive HIV-1 RNA test and other reasons. 10 out of 20 women became pregnant (50%), 8 women were on-going pregnant. The clinical and ongoing pregnancy rate per started cycle was 13% and 11% respectively. 7 babies have thus far been born and none of the mothers or babies seroconverted within this study period. Larger numbers of patients are necessary to support the safety of this program. PMID- 15751324 TI - [Publication practice and policy for disciplinary proceedings regarding health care in the Netherlands]. AB - OBJECTIVE: To provide insight into practice and policy regarding the publication of disciplinary verdicts in Dutch health care. DESIGN: Descriptive. METHOD: The study covered all of the verdicts issued by disciplinary boards and courts of justice that were published in the Dutch the Staats-courant (Government Gazette) during the period 1995-2002. The following attributes were recorded for each verdict: year of publication and verdict, the disciplinary board concerned, the accused professional, the type of complainant, the nature of the complaint and verdict, and the journals that were offered the decision. The published verdicts were related to the total number of verdicts and cases during the study period. Questionnaires were used to investigate the publication policy of the disciplinary boards. RESULTS: A total of 4% of all verdicts were published in the Dutch Government Gazette (323/8902). The central disciplinary board decided to publish more often than the regional disciplinary boards (8% and 2%, respectively). There were considerable differences between the various regional disciplinary boards (range: 1-5%). Per professional group the percentage of verdicts in cases that were published ranged from 2% to 23%. The decisions were offered to over 20 journals, but mainly to the Tijdschrift voor Gezondheidsrecht [Dutch Journal of Health Law] (TvGR; 92%) and Medisch Contact [Medical Contact] (MC; 88%). The TvGR published almost two-thirds of the verdicts offered (63%), and the MC published almost three-quarters (74%). In the decision to publish, the disciplinary boards differed in their interpretation of the concept of 'general importance'. CONCLUSION: If disciplinary proceedings are to achieve the desired quality-promoting effect on professional practice, then more attention will need to be paid to the publication policy, and the disciplinary boards will have to develop a joint code of practice. More verdicts could be published, also in professional journals. PMID- 15751325 TI - [Spirometry in patients with COPD in family practice and in a lung function laboratory equally reliable, but not interchangeable]. PMID- 15751326 TI - [The sensitization model: a method to explain chronic pain to a patient]. PMID- 15751327 TI - The cost-effectiveness of homeopathy: the perspective of a scientist and mother. PMID- 15751328 TI - Pharmacoeconomic comparison between homeopathic and antibiotic treatment strategies in recurrent acute rhinopharyngitis in children. AB - OBJECTIVES: A pharmacoeconomic study to compare, in terms of: medical effectiveness, quality of life and costs two treatment strategies ('homeopathic strategy' vs 'antibiotic strategy') used in routine medical practice by allopathic and homeopathic GPs in the treatment of recurrent acute rhinopharyngitis in 18-month to 4-year-old children. METHODS: Statistical analysis of data obtained from a population of 499 patients included in a previous 6-month prospective, pragmatic study. The patients were regrouped according to type of drug prescribed. Medical effectiveness was assessed in terms of (i) episodes of acute rhinopharyngitis, (ii) complications, (iii) adverse effects. Quality of life was assessed using the Par-Ent-Qol scale. Direct medical costs (medical consultations, drug prescriptions, prescriptions for further tests) and indirect medical costs (sick-leave) were evaluated from three viewpoints (society, patient, Social Security) using public prices and French Social Security tariffs. RESULTS: The 'homeopathic strategy' yielded significantly better results than the 'antibiotic strategy' in terms of medical effectiveness (number of episodes of rhinopharyngitis: 2.71 vs 3.97, P<0.001; number of complications: 1.25 vs 1.95, P<0.001), and quality of life (global score: 21.38 vs 30.43, P<0.001), with lower direct medical costs covered by Social Security (88 Euros vs 99 Euros, P<0.05) and significantly less sick-leave (9.5% of parents vs 31.6% of parents, P<0.001). CONCLUSIONS: Homeopathy may be a cost-effective alternative to antibiotics in the treatment of recurrent infantile rhinopharyngitis. PMID- 15751329 TI - Patients' assessments of the effectiveness of homeopathic care in Norway: a prospective observational multicentre outcome study. AB - OBJECTIVE: To evaluate the patient reported effects of homeopathic care 6 months after first consultations. METHODS: Prospective uncontrolled observational multicentre outcome study. All patients visiting 80 homeopaths all over Norway for the first time in eight different time periods from 1996 to 1998 were approached. Patients wrote down their main complaint and scored its impact on daily living on a 100 mm Visual Analogue Scale (VAS) at the first consultation. Six months later they were asked to score again. The homeopaths recorded treatments given for up to two follow-up consultations. MAIN OUTCOME MEASURE: Predefined as a reduction of at least 10 mm in the VAS score between the first consultation and follow-up. RESULT: Patients 1097 were recruited, 654 completed the follow-up questionnaire. The main complaint improved by at least 10mm on the VAS for 71% (95% confidence interval 67-74%) of patients. The average reduction was 32mm (95% CI 30-35 mm). Fifty-one per cent (95% CI 48-55%) of the patients had an improvement in their general well being of more than 10 mm. The mean reduction in the whole group was 14mm (95% CI 12-16 mm). The proportion of patients using conventional medication reduced from 39% to 16%. Regression analysis showed that lower age and higher baseline score were predictors of better outcome. CONCLUSION: In this study, seven out of ten patients visiting a Norwegian homeopath reported a meaningful improvement in their main complaint 6 months after the initial consultation. PMID- 15751330 TI - General practitioners and classical homeopaths treatment models for asthma and allergy. AB - Complementary and alternative treatment (CAT) is increasingly popular with patients and health-care providers. In the western societies an increasing number of patients with asthma and allergy combine conventional and CAT. But health-care providers knowledge about the different treatment models applied within the conventional health care system and alternative treatment is generally poor. 'Treatment model' includes understanding of disease, diagnostic approach, treatment methods and expected effects of treatment. Different treatment models represent different social constructions of understanding. We present a conceptualization of general practitioners' (GPs) and classic homeopaths' (CHs) perceptions of their treatment models in patients with asthma and allergy. This is a first phase of a research project conducted in Denmark to study prerequisites for bridge building between conventional and alternative therapists. The research questions were: How do the GPs and the CHs describe their treatment models for asthma and allergy? What are the differences and similarities between the treatment models? Is there a logical connection between the components of the GPs and CHs treatment models? Six GPs and 11 CHs participated in semi-structured interviews. The main difference is in the GPs and CHs descriptions of the purpose of treatment. Similarities are found in the perception of a successful treatment. Both the GPs and the CHs experience treatment effects to be dependent on contextual conditions. Differences and similarities between treatment models are discussed in relation to potential bridge building between conventional and alternative health care. PMID- 15751332 TI - Clinical homeopathic use of RNA: evidence from two provings. AB - Two homeopathic drug provings (HDPs) of RNA have been published. The methods are reviewed. Our method is double-blind and placebo-controlled, conforms to GCP and includes run-in and follow-up phases. Symptoms are selected from diaries according to predefined criteria. The results of the provings are compared with each other and with the homeopathic clinical use of RNA. At this stage it is difficult to draw conclusions concerning the clinical use of RNA. Further HDPs are required, they should follow GCP and standard, transparent protocols. PMID- 15751331 TI - Histopathological and immunophenotyping studies on normal and sarcoma 180-bearing mice treated with a complex homeopathic medication. AB - Canova is a homeopathic complex medicine, used as an immune modulator. We studied its effects in normal and sarcoma 180-bearing mice. Three control groups were also evaluated. The mice were examined at daily intervals and the tumours observed histologically. Peripheral blood was analysed by flow cytometry. A delay in the development, and a reduction in size of the tumours, and increased infiltration by lymphoid cells, granulation tissue, and fibrosis surrounding the tumour were observed with active treatment compared to control. All animals from the treated group survived, 30% of control groups died. In 30% of treated animals, a total regression of the tumour was confirmed using light microscopy, no regression was found in the control groups. Treatment with Canova increased total numbers of leukocytes and lymphocytes. Among lymphocytes, TCD4, increased in normal-treated group and B and NK cells in S180-treated groups. The results reflect enhanced immune response of the host after treatment with Canova. PMID- 15751333 TI - Richard Hughes Memorial Lecture 2004. Homeopathy, wholeness and healing. PMID- 15751334 TI - Models for explaining the homeopathic healing process: a historical and critical account of principles central to homeopathy. AB - The success of Homeopathy in curing many diseases has been a serious challenge to science. Nineteenth century explanations for the healing process of Homeopathy cannot withstand the scrutiny of modern science and need to be abandoned or modified. The surviving propositions are discussed. A biocybernetic model with multilevels of electromagnetic feedback loops offers a hope of explaining the healing process. This model, its explanation of the healing process and experimental support are elaborated. PMID- 15751335 TI - 20 years ago: the British Homoeopathic Journal, January 1985. PMID- 15751337 TI - Entanglement in provings. PMID- 15751336 TI - Interactions between specific and non-specific treatment effects. PMID- 15751338 TI - Nanoparticles and nanocapsules created using the Ouzo effect: spontaneous emulisification as an alternative to ultrasonic and high-shear devices. AB - The preparation of polymeric particles and capsules by means of spontaneous droplet formation and subsequent polymer precipitation or synthesis is well known. However, spontaneous emulsification is a phenomenon that has often been erroneously interpreted. This Minireview provides new insights into the preparation of metastable liquid dispersions by homogeneous liquid-liquid nucleation, and is based primarily on a recent study by Vitale and Katz (Langmuir, 2003, 19, 4105-4110). This spontaneous emulsification, which they named the Ouzo effect, occurs upon pouring, into water, a mixture of a totally water-miscible solvent and a hydrophobic oil--and optionally some water--thus generating long-lived small droplets, which are formed even though no surfactant is present. Herein, we review and reinterpret the most relevant publications on the synthesis of a variety of dispersions (pseudolatexes, silicone emulsions, biodegradable polymeric nanocapsules, etc.), which we believe have actually been synthesized using the Ouzo effect. The Ouzo effect may also become a substitute for high-shear techniques, which, to date have only been of limited utility on industrial scales. PMID- 15751339 TI - Design of molecular photonic wires based on multistep electronic excitation transfer. AB - Light-harvesting complexes, one of nature's supreme examples of nanoscale engineering, have inspired researchers to construct molecular optical devices, such as photonic wires, which are optimised for efficient transfer of excited state energy over large distances. The control parameters for the design and the advantages of single-molecule fluorescence spectroscopy for the study of such complex systems are discussed with respect to energy-transfer mechanisms, chromophore selection and arrangement as well as static and dynamic heterogeneity. PMID- 15751340 TI - "Hot spot" hydrocarbon oxidation catalysed by doped perovskites--towards cleaner diesel power. PMID- 15751342 TI - Infrared spectrum of the Au-C60 complex. PMID- 15751341 TI - Templating effect of lipid membranes on Alzheimer's amyloid beta peptide. PMID- 15751343 TI - Persistent methylbenzenium ions in protonated zeolites: the required proton affinity of the guest hydrocarbon. PMID- 15751344 TI - Morphology of frozen rare-gas layers. PMID- 15751345 TI - Reversible force generation in a temperature-responsive nanocomposite hydrogel consisting of poly(N-isopropylacrylamide) and clay. PMID- 15751346 TI - Femtosecond time-resolved geometry relaxation and ultrafast intramolecular energy redistribution in Ag2Au. AB - The ultrafast dynamics of the bimetallic cluster Ag2Au is investigated by pump probe negative ion-to-neutral-to-positive ion (NeNePo) spectroscopy. Preparation of the neutral cluster in a highly nonequilibrium state by electron detachment from the mass-selected anion, and subsequent probing of the neutral nuclear dynamics through two-photon ionization to the cationic state, leads to strongly probe-energy-dependent transient cation-abundance signals. The origin of this pronounced time and wavelength dependence of the ionization probability on the femtosecond scale is revealed by ab initio theoretical simulations of the transient spectra. Based on the analysis of underlying dynamics, two fundamental processes involving geometry relaxation from linear to triangular structure followed by ultrafast intramolecular vibrational energy redistribution (IVR) have been identified and for the first time experimentally observed in the frame of NeNePo spectroscopy under conditions close to zero electron kinetic energy. PMID- 15751347 TI - A DFT study of the adsorption and dissociation of CO on Fe(100): influence of surface coverage on the nature of accessible adsorption states. AB - In the present article, we report adsorption energies, structures, and vibrational frequencies of CO on Fe(100) for several adsorption states and at three surface coverages. We have performed a full analysis of the vibrational frequencies of CO, thus determining what structures are stable adsorption states and characterizing the transition-state structure for CO dissociation. We have calculated the activation energy of dissociation of CO at 0.25 ML (ML = monolayers) as well as at 0.5 ML; we have studied the dissociation at 0.5 ML to quantify the destabilization effect on the CO(alpha3) molecules when a neighboring CO molecule dissociates. In addition, it is shown that the number and nature of likely adsorption states is coverage dependent. Evidence is presented that shows that the CO molecule adsorbs on Fe(100) at fourfold hollow sites with the molecular axis tilted away from the surface normal by 51.0 degrees. The asorprton energy of the CO molecule is -2.54 eV and the C-O stretching frequency is 1156 cm(-1). This adsorption state corresponds to the alpha3 molecular desorption state reported in temperature programmed desorption (TPD) experiments. However, the activation energy of dissociation of CO(alpha3) molecules at 0.25 ML is only 1.11 eV (approximately 25.60 kcal mol(-1)) and the gain in energy is 1.17 eV; thus, the dissociation of CO is largely favored at low coverages. The activation energy of dissociation of CO at 0.5 ML is 1.18 eV (approximately 27.21 kcal mol(-1)), very similar to that calculated at 0.25 ML. However, the dissociation reaction at 0.5 ML is slightly endothermic, with a total change in energy of 0.10 eV Consequently, molecular adsorption is stabilized with respect to CO dissociation when the CO coverage is increased from 0.25 to 0.5 ML. PMID- 15751348 TI - Geometry optimization and conformational analysis of (C60)N clusters using a dynamic lattice-searching method. AB - A newly developed unbiased global optimization method, named dynamic lattice searching (DLS), is used to locate putative global minima for all (C6O)N clusters with Girifalco potential up to N=150. A simple greedy strategy is adopted for the basic frame, so DLS has a very high convergence speed and may converge at various configurations. As most structures are packed by basic tetrahedra, some sequences are defined by both configurations and the size of the basic tetrahedra. A sequence-based conformational analysis is carried out with the defined sequences by counting the hit number over 10,000 independent DLS runs for all the cases up to N = 5. It was found that the hit rate of a sequence is related to the size of the basic tetrahedra. U.e of this method proved that the Leary tetrahedral sequence is dominant in a certain range of cluster sizes, although the sequence has no potential energy advantage. The calculation results are also consistent with those of annealing experiments at high temperature, both in magic numbers and height of the peaks in the mass spectrum. PMID- 15751349 TI - One- and two-photon photochemistry and photophysics of poly(arylenevinylene)s containing a biphenyl moiety. AB - Photochemical and photophysical properties were investigated for poly(arylenevinylene)s containing a flexible biphenyl "hinge" unit by applying one-photon (OP) and two-photon (TP) excitation to explore excited-state properties. The poly(arylenevinylene)s were poly[(2,5-dihexyloxy-p phenylenevinylene)-alt-(4,4'-dihexyloxy-3,3'-biphenylenevinylene)] (1), poly[(2,5 dihexyloxy-p-phenylenevinylene)-alt-(2,2'-dihexyloxy-3,3'-biphenylenevinylene)] (2), and poly[(2,5-dihexyloxy-p-phenylenevinylene)-alt-(2,2' biphenylenevinylene)] (3). Effective emission quantum yields and related photonic properties were evaluated on a realistic per-chromophore basis using effective conjugation lengths based on the Strickler-Berg relationship. Intramolecular photocyclization was deduced to occur in the one case where the biphenyl molecular connectivity permitted the reaction, based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), heteronuclear multiple-quantum coherence (HMQC)-NMR, and gel-permeation chromatography (GPC) results. The various photoprocesses could be induced by either OP or TP excitation, though the first excited singlet state is the photoactive state. The higher excitation energy of the TP excited state favors indirect population of the S, state by electronic coupling between the TP and OP excited states [lambda(max)TPE (nm): 726; delta (GM): 1=229, 2=215, 3= 109). Photochemical processes occurring from the lowest OP excited state (S1) could therefore also be indirectly induced by TP excitation. PMID- 15751350 TI - Molecular adsorption-mediated control over the electrical characteristics of polycrystalline CdTe/CdS solar cells. AB - The effect of surface treatments on p-CdTe/n-CdS solar cell performance was examined. Adsorption of organic molecules with various magnitudes and directions of the dipole moment on p-CdTe resulted in controlled changes in electron affinity and surface bond bending. Similar adsorption on CdTe in state-of-the-art p-CdTe/n-CdS solar cells changes the cell performance, and we explain this by a combination of increased series resistance and changes in light absorption and in cell photovoltage. While at this stage no improvement in performance has been found with these cell structures, which are the result of years of empirical optimization, the molecular effect on the photovoltage shows that it is possible in this way to control the photovoltaic effect at this junction. Separate optimization may well lead to improvement by inserting a dipole layer near the photovoltaic interface. Our results also show that this is even possible when dipole adsorption is performed on the complete polycrystalline thin-film cell. PMID- 15751351 TI - The kinetics of Z/E isomerization of methyl oleate catalyzed by photogenerated thiyl radicals. AB - The reaction of beta-mercaptoethanol with methyl oleate in tertbutyl alcohol was investigated under photochemical conditions. The time-dependent Z/E isomerization of the unsaturated moiety as well as the thiol-adduct formation were analyzed on the basis of radical generation rates and established rate constants. This provides precise room-temperature rate constants for the reversible thiyl radical addition to both Z and E isomers. The rate constants for HOCH2CH2S* addition to the Z and E forms were found to be 1.6 x 10(5) and 2.9 x 10(5) M(-1) S(-1), respectively. For the beta-elimination of the thiyl radical from the alkyl radical adduct, the rate constants were 1.7 x 10(7) and 1.6 x 10(8) S(-1), the faster associated with the formation of the E isomer. DFT-BB1K calculations on 2 butene assign this large preference of fragmentation for the E isomer to a lower activation barrier for the formation of the E transition state from the equilibrium radical adduct structure. PMID- 15751353 TI - Phase separation of Lal3 inside single-walled carbon nanotubes. AB - Simple binary solids can be found to adopt unprecedented structures when confined into nanometre-sized cavities, such as the inner cylindrical bore of single walled carbon nanotubes (SWNT). In the case of the discussed Lal(x)@SWNT encapsulation composite, the Lal2 "crystal" fragment adopts the structure of bulk Lal3, with one third of the iodine positions unoccupied. A complete characterisation of the encapsulation composite was achieved using an enhanced digital restoration approach of high-resolution transmission electron microscopy (HRTEM) images. The resulting complex exit surface wave provides information about the precise structural data of both filling material and host SWNT, establishing the SWNT's chirality and thus enabling a prediction of the composite's overall electron-transport properties. PMID- 15751352 TI - Probing the N(5)-H bond of the isoalloxazine moiety of flavin radicals by X- and W-band pulsed electron-nuclear double resonance. AB - An X- (9.7 GHz and W-band (94 GHz) pulsed electron-nuclear double resonance (ENDOR) study of the flavin cofactor of Escherichia coli DNA photolyase in its neutral radical form is presented. Through proton and deuteron ENDOR measurements at T = 80 K, we detect and characterize the full anisotropy of the hyperfine coupling (hfc) tensor of the proton or deuteron bound to N(5) of the isoalloxazine ring. Scaling of the anisotropic proton hfc components by multiplication with the quotient of the magnetogyric ratio of a deuteron and a proton, chiD/chiH, reveals subtle differences compared to the respective deuteron couplings obtained by 95-GHz deuterium ENDOR spectroscopy on an H-->D buffer exchanged sample. These differences can be attributed to the different lengths of N(5)-H and N(5)-D bonds arising from the different masses of protons and deuterons. From the R(-3) dependence of the dipolar hyperfine splitting, we estimated that the N(5)-D bond is about 2.5% shorter than the respective N(5)-H bond. That such subtle bond-length differences can be resolved by pulsed ENDOR spectroscopy suggests that this method may be favorably used to probe the geometry of hydrogen bonds between the H(5) of the paramagnetic flavin and the protein backbone. Such information is only obtained with difficulty by other types of spectroscopy. PMID- 15751354 TI - On the low-lying excited states of sym-triazine-based herbicides. AB - We report a joint computational and luminescence study on the low-lying excited states of sym-triazines, namely, 1,3,5-triazine (1) and the ubiquitous herbicides atrazine [6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine (2)] and ametryn [6-methylthio-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine (3)]. Geometrical structures, energetics, and transition and state properties of I and 2 were computed at the TD-DFT, CASSCF, and CASPT2 levels of theory. The fluorescence and phosphorescence emission spectra, lifetimes, and fluorescence quantum yields were measured for the three compounds, and from these, the energies of the lowest excited states and their corresponding radiative rates were determined. The predictions from CASPT2 calculations are in good agreement with the experimental results obtained from the luminescence studies and allow the interpretation of different absorption and emission features. PMID- 15751355 TI - Determining the geometry of hydrogen bonds in solids with picometer accuracy by quantum-chemical calculations and NMR spectroscopy. AB - The structure of multiply hydrogen-bonded systems is determined with picometer accuracy by a combined solid-state NMR and quantum-chemical approach. On the experimental side, advanced 1H-15N dipolar recoupling NMR techniques are capable of providing proton-nitrogen distances of up to about 250 pm with an accuracy level of +/-1 pm for short distances (i.e., around 100 pm) and +/-5 pm for longer ones (i.e., 180 to 250 pm). The experiments were performed under fast magic-angle spinning, which ensures sufficient dipolar decoupling and spectral resolution of the 1H resonance lines. On the quantum-chemical side, the structures of the hydrogen-bonded systems were computationally optimised, yielding complete sets of nitrogen-proton and proton-proton distances, which are essential for correctly interpreting the experimental NMR data. In this way, nitrogen-proton distances were determined with picometer accuracy, so that vibrational averaging effects on dipole-dipole couplings need to be considered. The obtained structures were finally confirmed by the complete agreement of computed and experimental 'H and '5N chemical shifts. This demonstrates that solid-state NMR and quantum-chemical methods ideally complement each other and, in a combined manner, represent a powerful approach for reliable, high-precision structure determination whenever scattering techniques are inapplicable. PMID- 15751356 TI - Selective fluorescence recovery after bleaching of single E2GFP proteins induced by two-photon excitation. AB - We report the two-photon excitation and emission or a recently developed green fluorescent protein (GFP) mutant, E(2)GFP. Two main excitation bands are found at 780 and 870 nm. Blinking and irreversible and reversible bleaching were observed. Fluorescence blinking occurs in the millisecond range and has been ascribed to conversions between the neutral, anionic and dark zwitterionic states. Bleaching is observed after approximately 10 to 400 ms depending on the excitation power, and it is probably due to a conversion to a dark state. The striking feature of this GFP mutant is that the fluorescence can be recovered with very high efficiency only upon irradiation at 720 +/- 10 nm. This GFP mutant therefore seems promising as an almost permanent chromophore for two-photon excitation (TPE) microscopy or for applications in single-molecule memory arrays. PMID- 15751357 TI - Interactions between brushlike polyacrylic acid side chains on a polyacrylate backbone in dioxane-water. AB - Densely grafted polyacrylic acids (d-PAAs) with overcrowded PAA side chains on the polyacrylate main chains were synthesized and characterized. Acryloyl poly(tert-butyl acrylate) macromonomer [M-P(tert-BA)] was prepared with a definite chain length (n=29) by atom-transfer radical polymerization (ATRP), then homopolymerization was carried out to produce densely grafted P(tert-BA)s with polyacrylate main chains of two different lengths (m=27 and 161). The two d-PAAs were obtained by hydrolyzing d-P(tert-BA)s in the presence of trifluoroacetic acid (TFA). The d-PAAs exhibit intermolecular and intramolecular hydrogen bonding between the carboxylic groups of PAA side chains in dioxane and pyridine; both were investigated using proton nuclear magnetic resonance (1H NMR) spectroscopy. The intermolecular hydrogen bonding was found to be dependent on polymer concentration, temperature, and water content. The intramolecular association between the PAA side chains was found to produce a contraction of the hydrodynamic volume of the d-PAA. Intermolecular hydrogen bonding produces aggregates, as demonstrated by dynamic light scattering (DLS). The clusters were found to shrink as the overall water concentration decreased, and this effect is tentatively explained by considering the gradient in chemical potential of water inside the clusters in comparison with the solvent phase. PMID- 15751359 TI - Graphite powder and multiwalled carbon nanotubes chemically modified with 4 nitrobenzylamine. AB - We demonstrate that graphite powder and multiwalled carbon nanotubes (MWCNTs) can be derivatised by 4-nitrobenzylamine (4-NBA) simply by stirring the graphite powder or MWCNTs in a solution of acetonitrile containing 10 mM 4-NBA. We propose that 4-NBA partially intercalates at localised edge-plane or edge-plane-like defect sites and this hypothesis with a range of experimental data provided by electrochemistry in both aqueous and nonaqueous media, electron microscopy and X ray powder diffraction. PMID- 15751358 TI - A theoretical analysis of the coordination modes of CuII with penicillins: activation of the beta-lactam C-N bond. AB - The interaction of CuII with 6-formylamino-3alpha-carboxypenam and 6-acetylamino 3alpha-carboxypenam was investigated by means of DFT calculations with the UB3LYP functional. Nine different modes of complexation between CuII and 6-formylamino 3alpha-carboxypenam were located. When two water molecules directly bonded to CuII are included in the calculations on 6-acetylamino-3alpha-carboxypenam as penicillin model, only six CuII(H2O)2-6-acetylamino-3alpha-carboxypenam complexes (1S-6S) are found. In solution the four most stable complexes obtained from our calculations, 6S, 1S, 2S, and 3S, exhibit CuII in square-planar coordination with at least one bond to the carboxylate group, in agreement with experimental evidence. Complexes 6S, 1S, and 3S were previously suggested by available experimental evidence. In three of the most stable complexes (6S, 2S, and 3S) the beta-lactam C-N bond is remarkably activated and displays C-N bond lengths similar to those found in some tetrahedral intermediates located for the hydrolysis of 2-azetidinones. This suggests that these kinds of complexes belong to the reaction coordinate for the degradation of beta-lactam antibiotics in the presence of CuII. PMID- 15751360 TI - Absolute rate calculations: atom and proton transfers in hydrogen-bonded systems. AB - We calculate energy barriers of atom- and proton-transfer reactions in hydrogen bonded complexes in the gas phase. Our calculations do not involve adjustable parameters and are based on bond-dissociation energies, ionization potentials, electron affinities, bond lengths, and vibration frequencies of the reactive bonds. The calculated barriers are in agreement with experimental data and high level ab initio calculations. We relate the height of the barrier with the molecular properties of the reactants and complexes. The structure of complexes with strong hydrogen bonds approaches that of the transition state, and substantially reduces the barrier height. We calculate the hydrogen-abstraction rates in H-bonded systems using the transition-state theory with the semiclassical correction for tunneling, and show that they are in excellent agreement with the experimental data. H-bonding leads to an increase in tunneling corrections at room temperature. PMID- 15751361 TI - Your diagnosis? Scaphoid fracture. AB - Fractures of the scaphoid are the most common carpal bone fracture. With careful management, prognosis is excellent. Imaging is critical, not only for early diagnosis, but also for monitoring patients after initial treatment to detect unstable fractures, the common complications of nonunion and avascular necrosis, and the need for surgical treatment. PMID- 15751362 TI - Repetition makes a scholar. PMID- 15751363 TI - Minimally invasive treatment of simple bone cysts with percutaneous autogenous bone marrow injection. AB - Percutaneous autogeneic bone marrow injection is an effective method for managing simple bone cysts, and it might be considered before the application of more extensive procedures. PMID- 15751364 TI - Treatment of intertrochanteric hip fractures with the AO trochanteric fixation nail. AB - Further biomechanical and clinical studies are necessary to validate the efficacy of the Trochanteric Fixation Nail, but in our experience this is an improvement over the currently available devices. PMID- 15751365 TI - Once-daily aminoglycoside administration. AB - Once-daily aminoglycoside dosing is effective in treating gram-negative infections, including those associated with bones and joints. PMID- 15751366 TI - High-pressure injection injuries of the hand. AB - While often innocuous at presentation, high-pressure injection injuries can lead to devastating consequences. Stiffness, chronic pain, infection, and even amputation can occur, with amputation rates ranging between 16% and 48%. Early surgical decompression and debridement are the cornerstones of treatment. PMID- 15751367 TI - Current concepts in nonoperative management of knee osteoarthritis. PMID- 15751369 TI - The effects of weather and seasonality on hip fracture incidence in older adults. AB - This study examined the effect of weather and seasonality on hipfracture incidence in older adults residing in New York City. A total off 66,346 patients aged > or = 65 years who sustained a fracture of the femoralneck or intertrochanteric region from 1985 to 1996 comprised the study population. Hip fractures were more likely to occur in the winter than in any of the other seasons (P<.001). Factors significantly correlated with hip fractureincluded minimum daily temperature (r=.167, P<.001), daily wind speed (r=.166, P<.001), maximum daily temperature (r=.155, P<.001), minutes of sunshine (r=.067, P<.01), and average relative humidity (r=.033, P=.03). A greater number of hip fractures occurred in colder months, withambient temperature rather than any adverse circumstances related to rainor snowfall associated most closely to injury. As most fractures occurredindoors, precipitation is less likely to play a part in hip fracture occurrence in this population. PMID- 15751368 TI - Patient compliance with outpatient prophylaxis: an observational study. AB - The need for prophylaxis against venous thromboembolic disease following lower extremity joint replacement is well documented and accepted. Although low molecular weight heparin is approved for inpatient and extended prophylaxis beyond the hospital environment, orthopedic surgeons must determine the duration of injectable low molecular weight heparin based on the period of venous thromboembolic disease risk and on patient compliance with self-administration of medication at home. Lack of confidence in patients' ability to self-administer injections may contribute to hesitancy in prescribing injectable prophylaxis. This observational study evaluated the effectiveness of a self-injection education program. Findings demonstrate that when properly instructed, patients safely and efficaciously self-administer low molecular weight heparin at home. With shorter hospital stays, self-administration of low molecular weight heparin may optimize the clinical outcomes of orthopedic patients. PMID- 15751370 TI - The effects of lower-extremity total joint replacement for arthritis on obesity. AB - Total hip or knee replacement patients who are overweight or obese often consider their disabling joint disease a cause for their increased weight. This prospective study investigated weight change in 100 patients after successful total joint replacement to determine whether surgical treatment of hip or knee arthritis leads to weight reduction. Postoperatively, both hip and knee replacement patients gained weight, with no difference in weight gain between hip and knee replacement patients. Younger hip patients gained a significant amount of weight. Patients a with normal body mass index and obese patients did not lose weight, while overweight patients gained a significant amount of weight after surgery. These findings demonstrate successful treatment of lower-extremity arthritis does not lead to weight loss, and obesity should be treated as an independent disease that is not the result of inactivity from arthritis. PMID- 15751371 TI - Efficacy of platelet gel in reconstructive bone surgery. AB - Autologous platelet concentrate and cryoprecipitate, mixed to obtain a gel, have been successful in various operations, primarily oral and maxillofacial surgery. This study assessed the use of platelet gel in 19 patients undergoing 22 reconstructive bone surgical procedures. After a median follow-up of 12.9 months, improved osteoblastic reaction and reconstruction of physiologic bone structure was observed in all patients with no adverse reactions. These findings confirm the osteoinductive property of platelet gel in reconstructive bone surgery. PMID- 15751372 TI - Traumatic volar dislocation of the trapezoid with acute carpal tunnel syndrome. PMID- 15751373 TI - Bizarre parosteal osteochondromatous proliferation of the second metatarsal bone (Nora's lesion). PMID- 15751374 TI - Pseudowinging of the scapula due to subscapular osteochondroma. PMID- 15751375 TI - Septic arthritis of a lumbar facet joint associated with epidural and paraspinal abscess. PMID- 15751376 TI - Effect of the patient's own negligence. PMID- 15751377 TI - Physicians proudly serve and sacrifice. PMID- 15751379 TI - A grassroots "strike force" for Michigan health care. PMID- 15751378 TI - The politics of health care in Canada: visiting scholar offers unique insights. PMID- 15751381 TI - Health literacy and your practice. PMID- 15751382 TI - Valuing health care--the consequences to the workforce and to access. PMID- 15751380 TI - What information should be integrated with the childhood immunization registry? PMID- 15751383 TI - Bone formation in the maxillary sinus by using platelet-rich plasma: an experimental study in sheep. AB - Recently, platelet-rich plasma (PRP) has been proven to be an effective regeneration adjunct when combined with autogenous bone in the reconstruction of mandibular defects. However, little is known about the effect of PRP when combined with a bone allograft in the maxillary sinus. The purpose of this study was to quantitatively evaluate the ability of PRP to enhance bone regeneration in the maxillary sinus of sheep when combined with demineralized freeze-dried bone allograft (DFDBA) and cortical cancellous freeze-dried bone allograft (CCFDBA). Ten sheep were selected for bilateral sinus augmentation DFDBA + CCFDBA + PRP (test) and DFDBA + CCFDBA (control). Five were sacrificed at 3 months and the other 5 at 6 months. Hematology tests were performed for platelet count, and histology slides were obtained for histomorphometric analysis taking 2 measures of interest: total area (square millimeters) and percentage of bone fill. Student t tests showed no significant difference between test and control groups for total area (P > .25) and percentage of bone fill (P > .80) at either 3 or 6 months. The control group showed no statistical difference for total area (P < .095) and percentage of bone fill (P <.60) between 3- and 6-month healing times. The test group, however, showed a significant increase in total area (P <.025) but not in percentage of bone fill (P <.40) for the 2 healing periods. When the treatments were compared for interactions within the animal model, no clear tendency was evident for the test group to perform in relation to the control group regarding total area (r = .766, P < .01). A moderate tendency existed between the percentages of bone filled (r = .824, P < .005). Platelet-rich plasma showed higher platelet count than did the whole blood (2 to 5 times). However, no correlation was found between the log ratio and the bone measures. Within the limitations of this study, PRP failed to enhance or accelerate bone regeneration in the maxillary sinus of sheep when combined with bone allograft. PMID- 15751384 TI - Bone remodeling in immediately loaded and unloaded titanium dental implants: a histologic and histomorphometric study in humans. AB - Remodeling is thought to prevent microdamage accumulation caused by repetitive loading and to increase the fatigue life of bone. The bone remodeling rate (BRR) is the period of time needed for new bone to replace the existing bone and to allow for the adaptation of bone to its environment. BRR is expressed as a percentage or volume of new bone within a specific time period. The aim of the present study was to evaluate bone remodeling events on submerged and immediately loaded dental implants. Twelve patients with edentulous mandibles participated in this study. All patients were rehabilitated with fixed mandibular prostheses, with 10 dental implants per patient. An additional implant was inserted in the most distal posterior mandibular jaw region. In 6 patients, these additional implants were loaded with a fixed provisional prosthesis the same day of the implant surgery and loaded. In the other 6 patients, the additional implants were left submerged and not loaded. After 6 months, all the additional implants were retrieved with a trephine. The percentage of woven and lamellar bone, number of osteoclasts and osteoblasts, and percentage of bone labeled by tetracycline at 0.5 mm and 2 mm from the implant surface were evaluated. The percentage of lamellar bone, number of osteoblasts, and percentage of bone tetracycline labeling was significantly higher in the loaded implants than in the unloaded implants (P =.0001). Also in the loaded implants, the percentage of woven and lamellar bone, number of osteoclasts and osteoblasts, and percentage of bone tetracycline labeling was significantly higher at 0.5 mm than at 2 mm from the implant surface (P =.0001). No such differences were found in unloaded implants (P =.377). In conclusion, we found that (1) loading appeared to stimulate bone remodeling at the interface, (2) a higher percentage of lamellar bone was found in loaded implants, (3) the percentage of bone labeling was higher at the interface of loaded implants, (4) no differences were found in the BRRs between immediately loaded and unloaded implants, and (5) immediate loading had not interfered on the lamellar bone formation at the interface and had not produced formation of woven bone at the interface. PMID- 15751385 TI - 7-year follow-up of 93 immediately loaded titanium dental implants. AB - Recently, several experimental and clinical investigations found that immediately loaded implants obtained satisfactory levels of osseointegration with high success percentages. Only a few long-term studies of immediately loaded implants have been reported in the literature. The aim of this study was a 7-year clinical and radiographic follow-up of 93 immediately loaded dental implants in human patients. Eleven patients were consecutively enrolled in this study. A total of 7 full and 9 partial edentulous arches were rehabilitated. Patients presented a completely edentulous mandible (n=6), a completely edentulous maxilla (n=1), mandibular posterior edentulous areas (n=5), or a posterior maxillary edentulous area (n=1). Patients were rehabilitated with a bar and an overdenture (n=4), a provisional prosthesis of 3 to 12 elements (n=11), or a metal-ceramic bridge of 10 elements (n=1). A total of 93 implants were inserted and loaded within a 24 hour time frame. Six implants failed in the first year after loading. No more failures were observed in the following 6 years, and all the other implants were well integrated from a clinical and radiographic point of view. The cumulative success rate at 7 years was 93.5%, and the prostheses survival rate was 98.5%. The mean marginal bone loss was 0.6 mm after the first year and 1.1 mm at the 7 year evaluation. Primary stability is one of the most important parameters in immediately loaded implants because it avoids micromotion at the bone-implant interface. Four of the 6 failures in our patients occurred in partially edentulous patients; an excessive load applied to these small bridges could be the reason for the failure. Also, the bone quality is important, for 3 of our failed implants had been inserted in D3 bone. Our clinical and radiographic results have shown that these immediately loaded implants have remained osseointegrated for a long period. Our results point to the possibility of using the immediate loading technique in selected and well-informed cases. PMID- 15751386 TI - Procurement of autogenous bone from ramus with simultaneous the mandibular third molar removal for bone grafting using the Cr:YSGG laser: a preliminary report. AB - Autogenous bone grafting and third-molar removal are surgical procedures routinely performed in dentistry on a daily basis. The purpose of this preliminary report is to describe our clinical experience with the Er, Cr:YSGG laser in the procurement of bone harvested from the ramus and removal of third molars simultaneously from the mandible. PMID- 15751387 TI - Alveolar ridge augmentation: a comparative longitudinal study between calvaria and iliac crest bone grafrs. AB - Insertion of endosseous implants is often difficult because of lack of supporting bone. In the case of severe atrophy of the jaws, a large volume of autogenous bone can be harvested from the iliac crest and calvaria. Both grafts undergo partial resorption with time, but the rate of bone loss has not been fully elucidated. The aim of this study was to evaluate the alveolar bone height gain (ABHG) obtained with iliac crest and calvaria bone grafts. Twenty-five patients had mandibular bone grafts, 32 had maxillary bone grafts, and 11 had both mandibular and maxillary bone grafts. Measures were made on preoperative, postoperative, and follow-up radiographs. A general linear model was used to evaluate the rate of ABHG plotted against months elapsed from the time of the operation to the time of follow-up. General linear model output showed a statistically significant effect for only the type of donor bone graft (P =.004), with a better ABHG for calvaria. The iliac crest bone grafts lost most of the ABHG in the first 6 months, whereas calvaria bone grafts lost ABHG over a greater interval of time. The type of bone graft is the strongest predictor of ABHG, and calvaria bone graft had a higher stability than did iliac bone graft. However, the gap in ABHG between the 2 grafts tended to decrease over time. PMID- 15751388 TI - A proposal for universal nomenclature in implant prosthodontics. AB - Attempts have been made at formulating standardized nomenclature for implantology. Although these classification systems have advanced the concept of universal nomenclature in implantology, they can be improved upon. Most of them present terms in glossary form, which can limit their applicability. Others deviate significantly from accepted basic terminology and can be foreign or ambiguous to the average clinician. This article outlines the semiotic approach to language formulation, discusses slight changes to accepted conventional prosthodontic terminology to better encompass implant dentistry, and introduces the shortform and support-retention-connection-prosthesis classification systems. PMID- 15751389 TI - Cost assessment in sustainable waste management. PMID- 15751390 TI - Driving forces for import of waste for energy recovery in Sweden. AB - Between 1996 and 2002, the Swedish import of so-called yellow waste for energy recovery increased. The import mainly consisted of separated wood waste and mixes of used wood and paper and/or plastics that was combusted in district heat production plants (DHPPs). Some mixed waste was imported to waste incineration plants for energy recovery (10% of the import of yellow waste for energy recovery in 2002). The import came primarily from Germany, the Netherlands, Norway, Denmark and Finland. We identified six underlying driving forces for this recent increase of imported waste which are outlined and their interactive issues discussed. --The energy system infrastructure, which enables high energy recovery in Sweden. --The energy taxation, where high Swedish taxes on fossil fuels make relatively expensive solid biofuels the main alternative for base load production of district heat. --The quality of the waste-derived fuels, which has been higher in the exporting countries than in Sweden. --The bans on landfilling within Europe and the shortage of waste treatment capacity. --Taxes on waste management in Europe. --Gate fee differences between exporting countries and Sweden. In the future, the overall strength of these driving forces will probably be weakened. A Swedish tax on waste incineration is being investigated. In other European countries, the ambition to reach the Kyoto targets and increase the renewable electricity production could improve the competitiveness of waste-derived fuels in comparison with fossil fuels. Swedish DHPPs using waste-derived fuels will experience higher costs after the Waste Incineration Directive is fully implemented. The uncertainty about European waste generation and treatment capacity, however, might have a large influence on the future gate fees and thus also on the yellow waste import into Sweden. PMID- 15751391 TI - Construction waste management based on industrial management models: a Swedish case study. AB - This paper describes a methodology for estimating the true internal costs of construction waste, aimed at promoting environmentally friendly waste management. The study employs cost-benefit analysis, contribution margin analysis, the polluter-pays principle and a mathematical model: the model for Efficient Use of Resources for Optimal Production Economy (EUROPE), which has been introduced previously by the author for assigning industrial costs to waste. The calculations are performed on construction waste created in a case study of a building project. Moreover, waste is regarded as, in a business sense, having the same basic status as any normal industrial product, namely the 'equality principle'. Application of the methodology is suggested to create incentives for environmental and profitability improvement in construction companies and other types of industrial sectors. The results of the case study show the generation of construction waste to substantially decrease the final operating income, due to the internal shadow price cost it creates. This paper is intended to reduce the gap between the choice of waste management procedures and their economic impact, the overall objective being to accomplish an improved industrial environmental situation. PMID- 15751392 TI - The effect of food waste disposers on municipal waste and wastewater management. AB - This paper examines the feasibility of introducing food waste disposers as a waste minimization option within urban waste management schemes, taking the Greater Beirut Area (GBA) as a case study. For this purpose, the operational and economic impacts of food disposers on the solid waste and wastewater streams are assessed. The integration of food waste disposers can reduce the total solid waste to be managed by 12 to 43% under market penetration ranging between 25 and 75%, respectively. While the increase in domestic water consumption (for food grinding) and corresponding increase in wastewater flow rates are relatively insignificant, wastewater loadings increased by 17 to 62% (BOD) and 1.9 to 7.1% (SS). The net economic benefit of introducing food disposers into the waste and wastewater management systems constitutes 7.2 to 44.0% of the existing solid waste management cost under the various scenarios examined. Concerns about increased sludge generation persist and its potential environmental and economic implications may differ with location and therefore area-specific characteristics must be taken into consideration when contemplating the adoption of a strategy to integrate food waste disposers in the waste-wastewater management system. PMID- 15751393 TI - Comparative performance between temperature-phased and conventional mesophilic two-phased processes in terms of anaerobically produced bioenergy from food waste. AB - Comparative evaluation of bioenergy production from food waste was carried out with both a temperature-phased and a conventional mesophilic two-phased process at different organic loading rates (OLRs). No methane was detected in the temperature-phased thermophilic-acidogenic fermenter at all the OLRs tested. However, a significant amount of methane content was detected in the conventional two-phased mesophilic-acidogenic fermenter, with increments depending on the organic loading rate [from 17% at 3 g VS L(-1) day(-1) to 25% at 8 g VS L(-1) day(-1) (VS, volatile solid)]. Acetate and butyrate were the main volatile fatty acids (VFAs) in the temperature-phased thermophilic-acidogenic fermenter; conversely propionate was a major VFA in the conventional two-phased mesophilic acidogenic fermenter. Through the chemical oxygen demand (COD) balance of both temperature-phased and conventional mesophilic two-phased processes, the fraction of the feed-COD converted to the hydrogen-COD in the thermophilic-acidogenic fermenter within the former process was estimated from 7.9 to 9.3%, with a peak at ORL of 6 g VS L(-1) day(-1), whereas it was quantified from 0.3 to 0.9% in the mesophilic-acidogenic fermenter within the latter one. Moreover, the fraction of the feed-COD converted to the methane-COD in the mesophilic-acidogenic fermenter within the conventional two-phased process ranged from 5.4 to 7.9%. On the other hand, conversion of the feed-COD to the methane-COD in the mesophilic methanogenic fermenter of both temperature-phased and conventional mesophilic two phased processes ranged from 66.2 to 72.3% and from 63.5 to 70.5%, respectively, with decrements related to the increase of organic loading rate. PMID- 15751394 TI - Biotechnology for aerobic conversion of food waste into organic fertilizer. AB - A biotechnology for aerobic conversion of food waste into organic fertilizer under controlled aeration, stirring, pH and temperature at 55-65 degrees C, is proposed. To maintain neutral pH at the beginning of the bioconversion 5% CaCO3 was added to the total solids of the food waste. The addition of 20% horticultural waste compost as a bulking agent to the food wastes (w.w./w.w.), improved the bioconversion and increased the stability of the final product. No starter culture was needed for aerobic bioconversion of food waste into organic fertilizer for 10 days. The low contents of heavy metals in the raw materials used in the bioconversions ensured the safety of fertilizer from food waste for application in agriculture. The addition of 4% organic fertilizer to the subsoil increased the yield and growth of Ipomoea aquatica (Kang Kong) by 1.5 to 2 times. The addition of phosphorus is required to enhance the positive effect of organic fertilizer on plant growth. PMID- 15751395 TI - Microbial succession associated with organic matter decomposition during thermophilic composting of organic waste. AB - Using dog food as a model of the organic waste, thermophilic composting was carried out for 14 days at a fixed temperature of 60 degrees C. The relationship between organic matter decomposition measured by CO2 evolution during the bio stabilization process and microbial succession expressed as the changes over time in the restriction fragment length polymorphism (RFLP) patterns of 16S rDNA sequences, of micro-organisms associated with the composting material was also examined. The CO2 evolution rate peaked on day 3 and gradually decreased until it became extremely small after day 9 of composting, indicating that vigorous organic matter decomposition ceased around this time. On the other hand, the RFLP pattern changed drastically from day 0 to day 4 or 5, then remained stable until day 7 or 8, reaching its final configuration, with little variations, after day 9 of composting. RFLP analysis therefore indicates that microbial succession continued into the later stage of composting. Nevertheless, by day 9, the rate of organic matter decomposition was so low that its influence on microbial populations could be hardly recognized by conventional methods of dilution plating. Moreover, the compost produced by day 9 showed no inhibitory effect on the growth of Komatsuna (Brassica campestris L. var. rapiferafroug), indicating that the maturity of compost is sufficient for plant growth when the rate of organic matter decomposition has become extremely low and the RFLP patterns become stable. PMID- 15751396 TI - Chloride extraction for quality improvement of municipal solid waste incinerator ash for the concrete industry. AB - Coal ash from power stations has long been used successfully in the cement industry as binders in several Portland formulations. This is not the case for municipal solid waste (MSW) ash as chloride concentrations, ranging from 10 to 200 g kg(-1) dry weight in the bottom and fly ash, respectively, exceed the maximum allowable concentration in most cement mixtures. To reduce chloride content in MSW bottom ash, a laboratory investigation was carried out based on the exhaustive washing in tap water. The influence of operative parameters such as temperature, granulometric properties and solid/liquid ratio of extraction was evaluated. In addition to optimization of the mentioned operative parameters for full-scale application, the paper gives preliminary indications on mechanistic aspects of the washing operation. PMID- 15751397 TI - Characterization of solid waste disposed at Columbia Sanitary Landfill in Missouri. AB - Waste sorts were conducted during each of the four quarters (or seasons) of 1996 at the City of Columbia Sanitary Landfill. A detailed physical sampling protocol was outlined. Weight fractions of 32 waste components were quantified from all geographic areas that contribute to the Columbia Sanitary Landfill using a two way stratification method, which accounted for variations in geographical regions and seasons. Comparisons of solid waste generated between locations and seasons were conducted at the 80% confidence level. The composition of the entire waste stream was 41% paper, 21% organic, 16% plastic, 6% metal, 3% glass and 13% other waste. Paper was the largest composition and glass was the smallest composition for all geographical regions. The result of this study was also compared with a 1987 Columbia, Missouri study conducted by EIERA (1987), with studies conducted in other states such as Minnesota, Wisconsin, Oregon and with national study conducted by the USEPA (USEPA 530-R-96-001, PB96-152 160. US Environmental Protection Agency, Office of Solid Waste, Washington, DC). The results of studies from other states are different from this study due to different local conditions, different methodologies and a different scope. There was a small (5%) increase in per capita weight from 1987 to 1996. The total per capita weight in the present study was 60% greater than the national per capita weight reported by the USEPA (1996) due to that the USEPA report excluded industrial, construction and certain commercial waste. The total per capita weight agrees with the national per capita weight for municipal waste reported by Tchobanoglous (1993), which included industrial, construction and commercial sources. The geographical and seasonal effects on the waste composition are evaluated and discussed. Statistical analysis indicates that waste characteristics are different among geographical regions and seasons. The potential for waste recovery and reduction is also discussed. PMID- 15751398 TI - Factors affecting water balance and percolate production for a landfill in operation. AB - Percolate production and precipitation data for a full-scale landfill in operation measured over a 13-year period were used to evaluate the impact and importance of the hydrological conditions of landfill sections on the percolate production rates. Both active (open) and closed landfill sections were included in the evaluation. A simple top cover model requiring a minimum of input data was used to simulate the percolate production as a function of precipitation and landfill section hydrology. The results showed that changes over time in the hydrology of individual landfill sections (such as section closure or plantation of trees on top of closed sections) can change total landfill percolate production by more than 100%; thus, percolate production at an active landfill can be very different from percolate production at the same landfill after closure. Furthermore, plantation of willow on top of closed sections can increase the evapotranspiration rate thereby reducing percolate production rates by up to 47% compared to a grass cover. This process, however, depends upon the availability of water in the top layer, and so the evaporation rate will be less than optimal during the summer where soil-water contents in the top cover are low. PMID- 15751399 TI - Vaccinations for waste-handling workers. A review of the literature. AB - A review of the literature relating to the need for vaccination against infectious disease in the solid waste industry was conducted, focusing on hepatitis A, hepatitis B and tetanus. Databases (Medline, PreMedline, EMBASE, CINAHL, Current Contents, Cochrane Database, HTA Database, DARE, OSHROM) were searched up to and including August 2003. Articles were included in the review if they reported the prevalence of immunity to hepatitis A, hepatitis B or tetanus in solid waste workers or the incidence of clinical infection with any of these diseases. Papers about hazardous or medical waste, incineration or other infectious diseases were excluded. Forty-four papers constituted the evidence database. Only one paper studied the prevalence of antibodies to hepatitis A and hepatitis B in solid waste workers compared with sewage plant workers and office workers, and no difference was found between these groups of workers. There was some evidence to support a theoretical risk of infection with hepatitis A, B and tetanus; however, no studies could be found of the risk of these diseases in solid waste workers. No single cases of these diseases being acquired occupationally in solid waste management were identified in the literature. Workers in the solid waste industry may theoretically be at increased risk of acquiring infectious diseases occupationally. However, at present no studies could be found which have documented this risk. PMID- 15751400 TI - 'Stop pub': can banning of junk mail reduce waste production? AB - According to the new regulations on junk mail management to be adopted in France, both producers and distributors must participate in the recycling process. This case study was carried out in 2000 in four sectors in the Departement of Charente in the south west of France. This report presents the quantities of junk mail in residual waste and in separate collections. The conclusion is that the new regulations may reduce overall domestic waste production by 2-4%, with the impact on separately collected paper being some 20%. PMID- 15751401 TI - [Guidelines for the urological management of renal and ureteric stones in adults]. AB - Progress in the treatment of urinary stones over the last 20 years requires a clearer definition of the indications for each treatment modality for the urological management of renal and ureteric stones in adults. The Stones Committee of the Association Francaise d'Urologie reviewed the European and American guidelines, the literature published over the last five years, and the experience of its members to establish a set of practical guidelines for the treatment of urinary stones. The site (kidney or ureter) and size (dimensions) of the stone are the initial descriptive criteria for the choice of treatment. Indications are classified as "standard" for first-line treatments, or "optional" for alternative modalities. The success of treatment is defined by the absence of residual fragments. ESWL is the reference treatment for renal stones less than 20 mm in diameter. PCNL is the most effective treatment for stones larger than 20 mm. Flexible ureteroscopy is still under evaluation. Open surgery and laparoscopy have limited indications in less than 1% of cases. ESWL is the first-line treatment for proximal ureteric stones. ESWL and ureteroscopy are both treatments of choice for stones of the pelvic ureter less than 10 mm in diameter, while ureteroscopy is recommended for stones larger than 10 mm. Stone morphology and composition analysis and an aetiological survey must be performed after surgical treatment and the patient must be given dietary advice to prevent recurrence. PMID- 15751402 TI - [Terminology of lower urinary tract dysfunction: French adaptation of the terminology of the International Continence Society]. PMID- 15751403 TI - [Management of stage I nonseminomatous germ cell tumours after orchiectomy]. AB - BACKGROUND: Stage I nonseminomatous germ cell tumours are characterized by the absence of detectable metastatic extension at the time of diagnosis of the disease. The experience acquired with staging retroperitoneal lymphadenectomy or surveillance cohorts demonstrates that about 30% patients nevertheless present microscopic metastases. Should management after orchidectomy be based on watchful waiting or should complementary treatment be proposed immediately? METHODS: A review of the international literature was designed to: 1) evaluate the contribution to the decision-making process of histological variables able to predict the risk of micro-metastatic dissemination, 2) define the potential advantages and disadvantages of the three approaches usually adopted after orchidectomy (retroperitoneal lymphadenectomy, surveillance and adjuvant chemotherapy), 3) establish guidelines for daily practice. RESULTS: Histological variables are especially reliable for prediction of the absence of micro metastatic risk. The various approaches adopted after orchidectomy can achieve similar and excellent cure rates. The potential adverse effects are very different. Watchful waiting is preferable in low-risk patients (absence or small proportion of embryonic carcinoma and absence of vascular invasion in the primary tumour). Adjuvant chemotherapy must be strictly reserved to high-risk patients (predominant component of embryonic carcinoma and presence of vascular invasion). Retroperitoneal lymphadenectomy must be performed by specialized teams in patients at low or intermediate risk, especially when watchful waiting cannot be effectively ensured, when serum tumour markers are normal at diagnosis or when the primary tumour presents a predominant component of mature teratoma associated with vascular invasion. CONCLUSION: Histological data derived from the orchidectomy specimen, the urologist's learning curve in relation to retroperitoneal lymphadenectomy and the clinical context (patient's expected compliance with follow-up, expected adverse effects of the various treatment options) are the main decisional parameters to be taken into account by multidisciplinary teams. The optimal individual option can only be determined after discussion with the patient. PMID- 15751404 TI - [Therapeutic implications of the revised concept of hormone-refractory metastatic prostate cancer]. AB - Almost all patients with prostate cancer become refractory to hormonal therapy which blocks androgen-dependent cellular proliferation. The key to this resistance is related to overexpression of the androgen receptor (AR). Alternative methods designed to block signalling pathways mediated by AR appear to be critical for tumour survival. These signalling pathways, which interact with the AR, can increase the response to androgen deprivation. Identification of signalling pathways will be an important objective for the treatment of prostate cancer. Application of new treatments must be preceded by identification of the genetic and molecular profiles of each patient's tumour. PMID- 15751405 TI - [Blunt kidney trauma: a ten-year experience]. AB - OBJECTIVE: The objective of this study is to assess the results of our therapeutic management of blunt kidney trauma in patients hospitalised over the last 10 years. MATERIALS AND METHODS: From January 1993 to January 2003, 105 patients were hospitalised in our department for blunt kidney trauma. We retrospectively studied age, gender, injured side, mechanism of trauma (direct, indirect or deceleration), aetiology, presence of associated lesions (visceral, orthopaedic), and clinical and laboratory signs on admission (haematuria, blood pressure, haemoglobin and serum creatinine). The grade of the lesions was defined by radiological assessment, specifying the presence or absence of devascularized fragments and urine extravasation. All complications were noted and studied according to the initial therapeutic management and grade. Follow-up was clinical (BP and search for renal pain) and radiological (CT and/or DMSA scan). RESULTS: 105 cases of blunt trauma of the kidney were hospitalised between January 1993 and January 2003 in our department. The mean age of the patients was 28.7 years (range: 7-75 years). Trauma was classified into 5 grades on the basis of the radiological assessment according to the ASST (American Society of Surgery of Trauma): 51 (49%) cases of grade 1 (n = 26) and grade 2 (n = 25) trauma, and 54 (51%) cases of major grade 3 to 5 trauma: 17 grade 3 (16%), 28 grade 4 (27%) and 9 grade 5 (8%) were diagnosed. Among the cases of major trauma, 7 (13%) were operated urgently during the first 24 hours: 4 cases of grade 5 trauma with renal artery dissection and 3 cases of grade 4 trauma with immediate uncontrolled bleeding. The nephrectomy rate (partial and total), when major renal trauma (grade 3, 4 and 5) (n = 47) was managed conservatively was 23% (11 nephrectomies) with the loss of 9.5 renal units (20%); this rate was 57% for grade 4 trauma presenting urine extravasation and devascularized fragments (n = 14). Twelve patients (7 with grade 4 trauma and 5 with grade 3 trauma) were reviewed by DMSA scintigraphy with a mean follow-up of 63 months (range: 26-108 months). Traumatized kidneys presented a mean function of 41.8% (range: 26.4-50%). CONCLUSION: Blunt kidney trauma is usually managed conservatively. The development of interventional radiology, endourological drainage techniques and medical intensive care helps to maintain this attitude by decreasing the need for surgery, even in the most severe trauma. PMID- 15751406 TI - [Comparative analysis of conservative surgery versus radical nephrectomy for renal cancer]. AB - OBJECTIVES: To compare the results of conservative surgery (CS) and radical nephrectomy (RN) for the treatment of renal cancer. MATERIAL AND METHOD: Between 1988 and 1999, more than 900 patients were operated for renal cancer in our department. We compared results of CS (partial nephrectomy and/or lumpectomy) to those of RN on 2 groups of matched patients with a similar sample size (n=62) in terms of morbidity, course of renal function, oncological efficacy and survival. RESULTS: The mean follow-up was more than 5 years for the two groups. This series comprised a predominance of localized (pT1 and pT2) low-grade (1 and 2) tumours: 80%. In our experience, CS was responsible for greater morbidity than RN, essentially because of indications of necessity compared to elective indications. Although the local recurrence rate was significantly higher after CS, analysis of overall and specific survival did not reveal any significant difference between the two groups. CONCLUSIONS: Conservative surgery appears to be a reasonable alternative to radical nephrectomy for the treatment of renal cancer in some indications and allows nephron sparing. This option appears to be all the more justified in view of the increasingly frequent discovery of small tumours in increasingly young patients. The development of minimally invasive techniques, particularly laparoscopy, is another aspect of this "minimal" trauma approach, but must be shown to ensure an equivalent oncological efficacy. PMID- 15751407 TI - [Percutaneous nephrolithotomy (PCNL) in subjects over the age of 70: a multicentre retrospective study of 210 cases]. AB - INTRODUCTION: Percutaneous nephrolithotomy (PCNL) in subjects over the age of 70 is considered to be associated with a higher risk than extracorporeal shock-wave lithotripsy (ESWL). However this technique is sometimes necessary for very large or complex stones in patients with several comorbidities. STUDY OBJECTIVE: To evaluate the predictive factors of success and operative risks likely to influence the results of PCNL in a population of patients over the age of 70. MATERIAL AND METHOD: Retrospective study of 203 patients (110 males, 97 females) over the age of 70, in whom a total of 210 PCNL were performed over a 12-year period in ten referral centres for the treatment of stones. 68.5% of cases presented a comorbidity and the median ASA score was 2. The median stone dimensions were 24 x 15 mm. There were 67 solitary pelvic stones, 7 infrapelvic ureteric stones, 31 solitary inferior caliceal stones, 40 complex stones and 13 staghorn calculi. A standard one-stage operative technique was performed in 92% of cases; the nephrostomy tract was easily performed by the urologists themselves in 71.8% of cases. RESULTS: The overall stone-free (SF) rate was 70.8%. The patient's weight and height (p=0.03 and p=0.01), stone dimensions and their solitary nature were significant factors of success (p<0.00001 and p=0.01) with SF rates of 81.1% for pelvic stones and 90.3%for solitary inferior caliceal stones (p<0.003); the SF rate for complete staghorn calculi was only 30.8%. A history of stones (p=0.04) and diabetes (p=0.03) influenced the stone-free rate, but the other comorbidities, body mass index, and age did not influence the stone free rate. There were two deaths (0.9% and haemostasis nephrectomy was performed in 2 ASA 3 subjects in renal failure. In the other patients, there was no difference between preoperative and postoperative serum creatinine and haemoglobin levels. The ease of nephrostomy, the rapidity of PCNL, the day of removal of the drainage nephrostomy, the mean hospital stay (median stay: 6 days), sterility of postoperative urine and at the 1-month visit were significantly correlated with the SF rate. CONCLUSIONS: PCNL in subjects over the age of 70 years is a safe and reliable technique that achieved a stone-free rate of 70.8% for all types of stones combined. The best results were obtained for solitary stones larger than 20 mm in the renal pelvis or inferior calyx. Apart from diabetes, which remains a risk factor, well controlled comorbidities do not increase the operative risk. PCNL did not affect haemodynamic parameters or renal function. PMID- 15751408 TI - [Efficacy of extracorporeal lithotripsy in acute renal colic: prospective study about 57 consecutive patients]. AB - INTRODUCTION: This study was designed to evaluate the efficacy of ESWL in the management of acute renal colic. MATERIAL AND METHOD: From January 2003 to August 2003, 102 patients were hospitalised for renal colic. We treated 57 patients by ESWL using an EDAP LT 02 apparatus. Successful treatment was defined by stone fragmentation on the plain abdominal x-ray at 24 hours associated with resolution of pain after only one ESWL session. RESULTS: The overall complete success rate after only one ESWL session was 49%. Results according to site: UPJ stones: 46% of success, lumbar ureter: 14%, iliac ureter: 33%, pelvic ureter: 66%. According to size: < 5 mm: 57% of success, 6 to 10 mm: 70%, 11 to 15 mm: 27%, > 15 mm: 0%. No difference in success rate was observed according to the cumulative energy delivered or according to the duration of ESWL sessions. CONCLUSION: ESWL can be considered to be a first-line treatment for acute renal colic. Its best indications correspond to pelvic or UPJ stones less than 10 mm in diameter. We believe that it is useless to exceed a duration of 30 min per ESWL session at a frequency of to 2 Hz. PMID- 15751409 TI - [Composition of renal stones currently observed in non-industrialized countries]. AB - Up until relatively recently, renal stones in developing countries were considered to be very different from those observed in industrialized countries, essentially characterized by the predominance of phosphate and urate stones, while the predominant stones in industrialized countries are calcium oxalate stones. To verify whether this difference in the epidemiological profile is still observed today, we analysed renal stones collected in various regions of the globe and compared their composition to that of stones observed in France. MATERIAL AND METHOD: 1,042 stones were collected between 1991 and 2000 from 14 different countries or geographical zones: Sub-Saharan Africa (Cameroon, Mali, Senegal), North Africa (Algeria, Morocco, Tunisia), South America (Brazil, Paraguay), Asia Minor (Pakistan, Turkey), Far East (China, Laos, Vietnam) and French Polynesia (Tahiti). Stones were analysed by infrared spectrophotometry. The composition of these stones was compared to that of 24,706 stones collected in France over the same period and analysed according to the same protocol. RESULTS: Overall, the proportion of calcium oxalate stones was the same in adults in France and in developing countries (men: 75.7% contre 72%; women: 59.8% contre 56.3%), but was higher in children in non-industrialized countries (boys: 52.6% contre 31.8% in France; girls: 67.8% contre 48.8% in France, p<0.0001). The frequency of calcium phosphate stones was particularly low in boys in developing countries (8.3% contre 45.1% in France, p<0.0001) andfrequency of purine stones was higher in boys (21.3% contre 5.2% in France, p<0.0001) and in girls (13.6% contre 4.3% in France, p<0.05). Major differences were observed according to continent and region; struvite was present in 42.9% of stones in women in Sub Saharan Africa contre 13% in South America and 2.7% in Asia Minor. Purines were 4 times more frequent in Tahitian men than in North African men. Calcium phosphate stones were 10 times less frequent in men in Asia Minor than in the Far East. CONCLUSION: The epidemiology of renal stones is continuing to change all over the world towards a predominance of calcium oxalate stones, which is now generalized. Major differences in the frequency of the other constituents, particularly purines and struvite, reflect particular eating habits and infectious risk factors specific to certain population. PMID- 15751410 TI - [The ureteric risk in laparoscopic surgery]. AB - OBJECTIVE: To describe the circumstances of development, diagnostic approach and treatment of iatrogenic ureteric lesions after laparoscopic surgery. MATERIAL AND METHODS: From 1992 to 2003, nine patients with iatrogenic trauma of the ureter after laparoscopic surgery were managed by our team. RESULTS: The time to diagnosis ranged from 0 to 640 days. These injuries were related to gynaecological surgery in 56% of cases, gastrointestinal surgery in 22% of cases, vascular surgery in 11% of cases, and spinal surgery in 11% of cases. The two lesions identified intraoperatively were treated immediately with success. In the case of lesions identified secondarily, first-line endourological treatment was performed in 57% of cases with a good result without reoperation in only 25% of cases. Three patients required reoperation with 66% of satisfactory results and two patients were treated by nephrectomy. CONCLUSION: Intraoperative difficulties and limited experience of laparoscopic surgery are the factors most frequently identified in iatrogenic ureteric lesions. The time to diagnosis has a considerable impact on the modalities and results of management. PMID- 15751411 TI - [Use of MEOPA (nitrogen monoxide-oxygen mixture) as analgesic for prostate biopsies]. AB - OBJECTIVE: To determine the tolerance of prostate biopsies in the case of co administration of an analgesic gas composed of a ready for use mixture of nitrogen monoxide and oxygen (MEOPA) (marketed under the trade name of Kalinox), and intrarectal instillation of xylocaine gel by comparing the results with those obtained in a control group only receiving intrarectal xylocaine instillation. MATERIAL AND METHODS: Non-randomized, prospective study conducted on 100 cases, with a control group composed of the first 22 patients (before availability of MEOPA in the department), followed by 78 patients treated with MEOPA. A questionnaire was completed by the patient and by the outpatient nurse caring for the patient. RESULTS: A very significant reduction (p=0.003) of the VAS pain score (from 3.86 to 2.38) and a highly significant improvement (p<0.001) of global tolerance of the procedure based on the nurse's estimation of pain (VAS scale) were observed. The MEOPA group also presented a significant improvement of pain in patients who had previously undergone a series of prostate biopsies (p=0.043). The satisfaction rate was higher in patients receiving MEOPA, although 28.2% of patients experienced adverse effects, all minor and bothersome in only 3.81% of cases. CONCLUSION: The MEOPA analgesic method as a complement to intrarectal xylocaine instillation considerably improved tolerance of the procedure with no major adverse effects and without prolonging the procedure or the surveillance. It is simple to use, safe and effective. PMID- 15751412 TI - [Treatment of postoperative male urinary incontinence by INVANCE prosthesis: preliminary results]. AB - PURPOSE: To evaluate the preliminary results of treatment of postoperative male stress urinary incontinence (SUI) by the INVANCE (AMS) suburethral sling. MATERIAL AND METHODS: From June 2003 to May 2004, 22 patients with stress urinary incontinence secondary to prostatic surgery (13 radical prostatectomies, 7 Ablatherm, 2 transurethral resections of the prostate) were treated by INVANCE sling. The patients presented either grade 1 incontinence (1 to 2 pads per day), or grade 2 incontinence (3 to 4 pads per day), or grade 3 incontinence (5 or more pads per day). The INVANCE process uses a rectangular polyester sling positioned under the bulbar urethra via a perineal incision. It is attached to the 2 ischiopubic rami by 3 titanium screws using a disposable electric screwdriver. Patients were reviewed every 3 months to evaluate the results (continence, Qmax, post-voiding residual volume, IPSS score, quality of life) RESULTS: The mean duration of the procedure was 79 minutes (range: 60-120). No intraoperative incidents were observed. The bladder catheter was removed on D1 (n=20) or D2 (n=2). The mean length of stay was 3.3 days (range: 2-8). Complications consisted of perineal haematoma (n=1), acute urinary retention after catheter removal (n=4), sling infection requiring removal (n=1), and prolonged perineal pain (n=5). No de novo irritative or obstructive urinary tract disorders were observed. With a median follow-up of 12 weeks (range: 3-44), 11 patients were completely continent (50%), 5 patients were significantly improved (22.7%) and 6 patients were considered to be failures (27.3%), 3 of whom had previously received external beam radiotherapy. CONCLUSION: The INVANCE process appears to be a simple and effective option for the treatment of grade 1 and 2 postoperative male stress urinary incontinence. PMID- 15751413 TI - [Sexual function after radical prostatectomy does not affect global patient satisfaction]. AB - OBJECTIVES: To evaluate the quality of life after retropubic radical prostatectomy (RP) and its impact on global patient satisfaction concerning the treatment received. PATIENTS AND METHODS: 142 questionnaires were sent to patients treated for a localized prostate cancer by RP alone, with a minimum follow-up of 2 years. The questionnaire was the validated French version of the "UCLA-Prostate Cancer Index". A question concerning global patient satisfaction with treatment was added. RESULTS: 102 questionnaires were returned and analysed. The mean age of the patients at the time of RP was 63.8 years and the mean follow up was 48 months. Evaluation of global satisfaction showed that 35/102 (343%) patients were very satisfied, 45/102 (44.1%) were satisfied, 15/102 had no opinion, 4/102 (3.9%) were dissatisfied and 3/102 (29%) were very dissatisfied. General quality of life scores ranged from 72 to 87 on a scale from 1 to 100 (where 100 corresponds to the best quality of life). For specific quality of life, the mean scores for sexual function and dysfunction were 27.5 and 25.1, respectively. The mean scores for urinary function and dysfunction were 72.5 and 67.8, respectively. Urinary function scores and seven of the nine general quality of life items were significantly correlated with better global patient satisfaction, but sexual function was not related to global satisfaction. CONCLUSIONS: Although sexual function is markedly altered after RP, it does not affect global patient satisfaction with the treatment received. Urinary function and general quality of life are significantly correlated with global satisfaction. PMID- 15751414 TI - [Design of a prototype operating seat with SESAM (Ergonomic System of Mobile Forearm Rests) mobile armrests designed to optimize the surgeon's ergonomy during pelvic laparoscopy]. AB - INTRODUCTION: Pelvic laparoscopic surgery offers multiple advantages for patients, but requires uncomfortable, non-ergonomic positions for the surgeon. Only a remote operator (Robot "slave" reproducing the surgeon's movements performed on a "master" console situated away from the patient), like Da-Vinci or Zeus (Intuitive Surgical) can improve the laparoscopic surgeon's working conditions. The objective of this study, based on an ergonomic analysis and recording of a laparoscopic surgeon's movements, was to define the specifications for the manufacture of an operating seat with armrests, based on the surgeon's position using a remote operator. MATERIAL AND METHODS: Movements of the centre of pressure and upper limbs of 12 urologists and gynaecologists with various levels of experience, were recorded using a force platform and a SAGA 3 RT movement analysis system (Biogesta), during 4 exercises performed on a pelvi trainer, with the surgeon positioned to the left (introduction of a needle, unravelling of a cord, dissection, suture). Ergonomic analysis of the laparoscopic surgeon's movements was based on video recordings of a surgeon under real operating conditions. The specifications were defined from all of these data. RESULTS AND DISCUSSION: The small amplitude of movements of the surgeon 's centre of pressure (< 45 cm2) confirmed that surgeons can be seated during laparoscopy. Recordings of the upper defined the elbow working zones and the need for 3D mobile armrests. Ergonomic analysis of posture defined the characteristics of the prototype. On the basis of these specifications, a prototype operating seat was developed. This prototype now needs to be validated or invalidated clinically. PMID- 15751415 TI - [Retroperitoneal haematoma and retropubic suburethral TVT tape. Report of two cases]. AB - The authors report two cases of retroperitoneal haematoma following insertion of retropubic suburethral TVT tape. In the first case, haemodynamic instability required emergency admission to the intensive care unit. Due to persistent haemodynamic instability, interventional radiology was performed to localize and embolize the vessel responsible for the bleeding, resulting in stabilization of the patient's haemodynamic state. In the second patient, blood loss was well supported allowing simple surveillance of the size of the haematoma, which remained stable, and correction of anaemia. Significant vascular wounds are rare after TVT and may require multidisciplinary management. Information appears to be essential, particularly for patients treated on an outpatient basis, to avoid delayed management. PMID- 15751416 TI - [Chronic TVT infection: mechanisms and consequences]. AB - Despite the simplicity and efficacy of TVT, this technique, like all surgical treatments, is nevertheless associated with sometimes very serious complications. The authors report a case of chronic infection of a polypropylene implant presenting with cutaneous fistula of a retropubic abscess without an associated septic syndrome, ten months after TVT insertion. The TVT was removed by mini laparotomy after failure of three weeks of adapted antibiotics and local wound care. These infections can be explained by the capacity of certain micro organisms to bind to biomaterials and produce a biofilm, protecting from the action of antibiotics and immune cells. PMID- 15751417 TI - [Bipolar loop resection of a bladder tumour in a pregnant woman]. AB - The authors report the case of a three-month pregnant woman, in whom a bladder tumour was discovered incidentally by ultrasound, raising the question of the possible foetal repercussions of using an electric scalpel in cutting mode. The bladder tumour was resected using a bipolar electric scalpel and irrigation was performed with physiological saline. No intraoperative or postoperative foetal repercussions were observed. Delivery was conducted with no particular problems and the newborn infant was perfectly healthy. The bipolar electric scalpel is not used routinely, but could be useful in pregnant women or patients with a pacemaker. PMID- 15751418 TI - [Leiomyoma of the female urethra]. AB - The authors report a case of leiomyoma of the female urethra and review the literature concerning this disease. This is a rare tumour that only exceptionally involves the urethra and is therefore difficult to diagnose. Urethral leiomyoma can present in the form of urinary tract infections, detrusor instability, haematuria, dyspareunia, and, more rarely, dilatation of the proximal urinary tract. Treatment is surgical. There is no risk of metastases, but local recurrence can occur in the case of incomplete resection. PMID- 15751419 TI - [Uroflowmetric functional evaluation of modified Duplay procedure in hypospadias surgery]. AB - We perform a modified Duplay procedure in our department without incision of the urethral plate for correction of distal hypospadias. The objective of this retrospective study was to analyse the long-term functional and aesthetic results of this technique. Strictures are a frequent complication of hypospadias surgery, but their frequency is often underestimated as they can remain asymptomatic. We decided to systematically detect urethral strictures by performing voiding uroflowmetry in all children over the age of 4 years, i.e. toilet trained, operated by Duplay procedure in our department. The results were compared to the standard results in paediatric populations published in the literature. The maximum flow rate was below the 5th percentile in 20% of cases and the average flow rate was abnormal in 30% of cases. Voiding uroflowmetry is a simple, noninvasive method to detect asymptomatic urethral strictures. Uroflowmetry is essential to evaluate the functional results of surgical procedures used for the treatment of hypospadias. PMID- 15751420 TI - [Early metastatic dissemination after nephro-ureterectomy for upper urinary tract tumour: a possible role of laparoscopy?]. AB - Laparoscopic nephro-ureterectomy has been proposed as an alternative to conventional double-incision surgery to treat upper urinary tract tumours. A case of hepatic and pulmonary metastatic dissemination four months after laparoscopic nephro-ureterectomy for pT3 N0 M0 intra-renal urothelial carcinoma raises the question of the possible role of laparoscopy in the rapidly unfavourable course of the disease. This case is discussed in the light of a review of the literature. PMID- 15751421 TI - [Natural history of a suspicious renal mass in a patient with gastric cancer: how to evaluate the risk of multiple primary tumours (MPT)?]. AB - The coexistence of several tumours in the same patient is a rare phenomenon and corresponds to the multiple primary tumour (MPT) syndrome. The most frequently diagnosed MPT include gastrointestinal tumours associated with urological tumours. Overexpression of the STAG1 gene could explain the development of several solid tumours and could be the origin of the multiple primary tumour syndrome. A suspicious renal mass discovered at the same time as a documented cancer is usually a malignant lesion and may justify combined radical surgical treatment. The authors report the natural history of a suspicious renal mass in a patient with primary gastric tumour corresponding to a MPT syndrome. They recall and discuss the epidemiology, pathophysiology, diagnosis and treatment of MPT. PMID- 15751422 TI - [Urinary bladder mesonephric clear cell adenocarcinoma]. AB - Mesonephric adenocarcinoma of the urinary bladder is a rare tumour, whose origin is still uncertain. The authors report the case of a 19-year old woman hospitalized for a huge pelvic mass revealed by hematuria. Trans-urethral biopsy showed the mesonephric nature of this mass. The treatment consisted of an anterior pelvectomy with total colpectomy, associated to complementary radiotherapy. With a follow-up of 30 months, the patient is still alive without recurrence or metastasis. The etiopathogenic, histologic and therapeutic aspects of this tumour are reviewed. PMID- 15751423 TI - [Inflammatory myofibroblastic tumours of the bladder]. AB - The inflammatory myofibroblastic tumour of the bladder is a rare benign affection that interests mainly young adults. Its etiopathogeny remains unknown, but its tumoral origin was evocated recently by Griffin (1999), incriminating a chromosomic abnormality involving the ALK gene. We will discuss the etiopathogenic, anatopathological and therapeutical aspects of this lesion for which the diagnosis is histological and the treatment remains conservative with a good prognosis. PMID- 15751425 TI - [Recto-seminal fistula and cancer of the rectum]. AB - The authors present a case of left epididymo-orchitis associated with rectal adenocarcinoma and suspected fistula between the left seminal vesicle and the rectum. This fistula was confirmed by pelvic computed tomography and surgical exploration. Treatment consisted of parenteral antibiotics and surgical drainage of the abscess associated with colorectal resection. This is an original case, as, retrospectively, the cancer of the rectum, diagnosed 4 days previously, was found to be responsible for the recto-seminal fistula and the seminal vesicle abscess. PMID- 15751424 TI - [Combined traumatic rupture of the bladder and ileum]. AB - Intraperitoneal bladder rupture represents 2% of all abdominal lesions requiring surgical exploration. It has a poor prognosis due to the frequent associated lesions, particularly involving abdominal and retroperitoneal organs. An associated small bowel injury is rarely reported in the literature. The limited intestinal symptoms, the poor sensitivity of emergency imaging and the minor lesions detected at surgical exploration justify a systematic search for ileal lesions in this type of trauma, by an incision providing broad exposure of the entire abdominal cavity. PMID- 15751426 TI - [Non-Hodgkin's malignant lymphoma: a rare cause of testicular tumour]. AB - The authors report a clinicopathological case of non-Hodgkin's malignant lymphoma of the testis in a 66-year-old man. The diagnosis was established on a right orchidectomy specimen. Conventional histopathological examination and immunohistochemistry concluded on diffuse large B-cell lymphoma. Staging showed involvement of Waldeyer's ring and the presence of retroperitoneal and para aortic deep lymphadenopathy. PMID- 15751427 TI - [Radical prostatectomy with transurethral dissection of the prostate and seminal vesicles]. AB - Transurethral dissection of the prostate and seminal vesicles was performed as the first step of suprapubic radical prostatectomy. Dissection was performed with a two-channel resector. Release of the anterior surface of the seminal vesicles and ampullae of the vas deferens was started at the bladder neck, in an infratrigonal plane. Partial section of the urethra was then performed proximally to the striated sphincter under visual and digital control to release the posterior surface of the prostate and seminal vesicles. Transurethral preparation of radical prostatectomy is technically feasible on the basis of our brief experience. This type of endoscopic prostatovesicular dissection avoids traction on the prostate, and theoretically decreases the risk of damage to the neurovascular pedicles (erection preserved in 6 out of 10 patients). Similarly, endoscopic section of the urethra theoretically decreases traumatic lesions of the striated sphincter (continence after removal of the bladder catheter in 9 out of 10 patients). The oncological result and operating time were comparable to those of conventional suprapubic radical prostatectomy. The risk of absorption is negligible in our experience. A large series is obviously necessary to confirm whether this technique constitutes a real improvement, particularly in terms of preservation of continence and erection. PMID- 15751428 TI - [A little known problem in schoolgirls: urinary tract infection and voiding disorders in young girls]. PMID- 15751429 TI - [Antonin Jean Desormeaux]. AB - Antonin Jean Desormeaux was the first to regularly perform optical examination of the male urethra. He advanced the knowledge of urethral disease and the treatment of urethral strictures and paved the way for the development of endoscopy. PMID- 15751430 TI - Where next for mental health? PMID- 15751431 TI - Invisible knowledge. PMID- 15751432 TI - Breakthrough in epilepsy care. PMID- 15751433 TI - Tsunami aftermath. PMID- 15751434 TI - Nurses decision-making in clinical practice. AB - AIM: To identify what decisions nurses make in medical, surgical and critical care areas and compare the results. METHOD: A clinical decision-making questionnaire (CDMQ) consisting of 15 statements was developed. A total of 60 nurses completed the questionnaire: 20 from each of three clinical areas. RESULTS: Most nurses, in all specialties, regularly made clinical decisions on direct patient care, which included providing basic nursing care and psychological support, and teaching patients and/or family members. Although nurses in all specialties regularly managed the work environment, they did not make decisions on the ward or unit budget, supervise junior staff or mentor student nurses. Critical care nurses regularly made decisions on their extended roles, such as acting in emergency situations and deciding to change patient medication, while medical and surgical nurses only did this occasionally. Length of clinical experience is significantly related to the frequency of decision making. CONCLUSION: The decisions nurses make are directly related to the clinical areas in which they work. However, it would be interesting to know if nurses showed particular aptitudes for different types and levels of decision making and if this is related to other factors such as personality, education and experience in nursing. PMID- 15751435 TI - The role of modern matrons in infection control. AB - This article discusses the effect of the role of the modern matron on healthcare associated infection (HCAI) and hospital cleanliness. The way the modern matron's role is developing is examined in relation to HCAI in a large acute NHS trust. As set out in Implementing the NHS Plan: Modern Matrons (Department of Health (DH) 2001), the role of modern matron includes a responsibility to lead clinical teams in the prevention of HCAI. PMID- 15751436 TI - Addressing issues related to adult patients who lack the capacity to give consent. AB - Capacity to consent is an area of practice that raises many ethical issues. The assessment by practitioners requires a flexible and creative approach. The Mental Capacity Bill is under final scrutiny by the House of Lords' committee and is intended to provide clearer guidance to focus practice in the future. PMID- 15751438 TI - New rules for safe practice. PMID- 15751437 TI - Prescription drugs: uses and effects. Thyroid disorders: symptom control. PMID- 15751439 TI - 'The prairie tortoise'. PMID- 15751440 TI - Domestic violence: recognising the signs. PMID- 15751441 TI - Record keeping--just for the fun of it? PMID- 15751442 TI - Improving outcomes in children with disability. PMID- 15751443 TI - Minor illness management: empowering parents through shared knowledge. PMID- 15751444 TI - Caring for young people with eating disorders in the acute medical setting. PMID- 15751445 TI - Removing k-wires: an audit of practice. PMID- 15751446 TI - Detecting and managing deterioration in children. PMID- 15751447 TI - Validity and reliability--what's it all about? Part 3: Issues relating to qualitative studies. PMID- 15751448 TI - Safeguarding children. AB - Nurses are well placed to identify behaviour or circumstances that might signify a child is at risk and to contribute to measures designed to protect children. Legislation, guidance and local policies provide the framework for nurses to implement good practice in communication, documentation and multi-agency working to safeguard every child and young person with whom they have contact. PMID- 15751449 TI - Treatment of chronic pain in persons with dementia: an overview. AB - The purpose of this review is to identify the common causes of nonmalignant chronic pain in people with dementia, discuss methods for determining the level of pain in patients who are nonverbal, and evaluate pharmacologic treatment for nonmalignant chronic pain. PMID- 15751450 TI - Integrated treatment approach improves cognitive function in demented and clinically depressed patients. AB - The purpose of this study was to evaluate the efficacy of an integrative treatment approach on cognitive performance. The study sample comprised 35 medically ill patients (20 male, 15 female) with an average age of 71.05, who were diagnosed with mild dementia and depression. These patients were evaluated at baseline and at six, 12, and 24 months of treatment, which included antidepressants (sertraline, citalopram, or venlafaxine XR, alone or in combination with bupropion XR), cholinesterase inhibitors (donepezil, rivastigmine or galantamine), as well as vitamins and supplements (multivitamins, vitamin E, alpha-lipoic acid, omega-3 and coenzyme Q-10). Patients were encouraged to modify their diet and lifestyle and perform mild physical exercises. Results show that the integrative treatment not only protracted cognitive decline for 24 months but even improved cognition, especially memory and frontal lobe functions. PMID- 15751451 TI - Evaluating hospital care for individuals with Alzheimer's disease using inpatient quality indicators. AB - The purpose of this study was to determine whether persons with Alzheimer's disease (AD) were at greater risk for in-hospital mortality than non-AD patients as a result of poor quality of care. The study focused on six common medical conditions that result in hospital mortality. Using 1995 to 2000 data from New York state (n = 7,021,065), analysts compared mortality risk for individuals with and without AD. Among men, adjusted odds of death were greater for those with AD for gastrointestinal (GI) hemorrhage (+52 percent), congestive heart failure (CHF) (+42 percent), hip fracture (+35 percent), and acute myocardial infarction (AMI) (+30 percent) (all p < .0001). Among women, AD did not affect risks for most conditions. The results of the study show that men with AD are at higher risk of hospital mortality for common medical conditions, which may indicate poor quality of care. Their risk of hospital death was greater than that of men without AD for AMI, CHF, hip fracture, and GI hemorrhage. Their risk was also greater than that of women with AD for CHF, pneumonia, hip fracture, and GI hemorrhage. With the exception of pneumonia, this risk difference notably exceeded the analogous difference between women and men without AD. Hospital staff should be alerted to greater mortality risk for men with AD, as this risk may indicate lower quality of care. PMID- 15751452 TI - Assessment of active music participation as an indication of subsequent music making engagement for persons with midstage dementia. AB - Persons with midstage dementia and in special care (N = 45) were assessed in groups by a music therapy practitioner to determine the level of engagement in a 15-minute protocol that included a five-minute segment for each of three music activity types--rhythm playing, exercising with music, and singing. Activity staff with little to no formal music training who were employed by the facility were taught to use the protocol to conduct eight subsequent activity sessions for small groups from which activity engagement data were collected for each subject. Results indicated the protocol was accessible and successful for indigenous activity staff, initial assessments were strong predictors of subsequent engagement, and participation levels were stable over time and across each of the three activities. PMID- 15751453 TI - Development of the knowledge about memory loss and care (KAML-C) test. AB - The purpose of this study was to develop a test to assess the knowledge of family caregivers concerning memory loss, the early stages of Alzheimer's disease (AD), and related care issues. A total of 31 multiple-choice items were generated in three domains: medical information, caregiving, and legal/financial planning. The test was administered to experts in the field of dementia research and care, medical students, and family caregivers. Item analysis procedures were then used to reduce the test to 15 items. Results suggest that the Knowledge about Memory Loss and Care (KAML-C) test is a reliable and valid test for examining the knowledge level of family caregivers with a relative in the early stages of dementia. PMID- 15751454 TI - Data and the void. PMID- 15751455 TI - Current management of congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a medical conundrum that challenges the physicians who care for these patients. Despite early prenatal diagnosis and optimal medical management, the results are disappointing. This lack of consistent success leaves the clinician frustrated. We present the current status of fetal surgery and novel approaches to ventilation as well as other unusual therapeutic approaches. Two cases are presented and a summary of our 11-year experience with CDH is reviewed. We conclude with complications and long-term outcomes. PMID- 15751456 TI - Prevalence of illicit drug use among individuals with chronic pain in the Commonwealth of Kentucky: an evaluation of patterns and trends. AB - The abuse of prescription-controlled substances is a common phenomenon, associated with illicit drug use, in patients suffering with chronic pain and receiving controlled substances for the management of pain. Prescription drug abuse in Kentucky has led to an increase in Medicare and Medicaid fraud, as well as physician indictments for unscrupulous practices and increased vigilance and prosecution by the authorities. This study was undertaken to evaluate 400 consecutive patients receiving controlled substances in an interventional pain management practice in Western Kentucky to evaluate the prevalence of illicit drug use and opioid abuse and noncompliance with opioid therapy. Results indicated that patients covered by third-party insurance showed 17% (95% CI, 10% 24%) prevalence of illicit drug use, with patients on Medicare, with or without third-party insurance, showing 10% (95% CI, 4%-6%) prevalence, compared to patients on Medicare and Medicaid with 24% (95% CI, 16%-32%) prevalence, and patients with only Medicaid with 39% (95% CI, 29%-49%) prevalence of illicit drug use. In addition, combined use of illicit drugs and misuse of prescription drugs was also seen in a substantially greater proportion of patients in Groups III and IV, with the highest (60%) in Group IV with Medicaid only, followed by Group III with Medicaid supplementation of Medicare insurance in 40% of the patients. Overall illicit drug use, opioid abuse, and noncompliance of opioids is significant in patients in Western Kentucky receiving Medicaid as a primary insurance or as a supplemental insurance to Medicare, despite extensive monitoring, written agreement, and education. PMID- 15751457 TI - Mysteries. PMID- 15751458 TI - Setting a good example. PMID- 15751459 TI - A shot in the dark. PMID- 15751460 TI - Challenging delays in thrombolysis. PMID- 15751461 TI - Board's eye view. PMID- 15751462 TI - Practical benchmarking. PMID- 15751463 TI - Lost generation? PMID- 15751464 TI - Keeping an open mind on mental health. PMID- 15751465 TI - Using a risk assessment matrix with mental health patients in emergency departments. PMID- 15751466 TI - Spinal cord injury: causation and pathophysiology. PMID- 15751467 TI - Tactile selective attention and body posture: assessing the multisensory contributions of vision and proprioception. AB - This study addressed the role of proprioceptive and visual cues to body posture during the deployment of tactile spatial attention. Participants made speeded elevation judgments (up vs. down) to vibrotactile targets presented to the finger or thumb of either hand, while attempting to ignore vibrotactile distractors presented to the opposite hand. The first two experiments established the validity of this paradigm and showed that congruency effects were stronger when the target hand was uncertain (Experiment 1) than when it was certain (Experiment 2). Varying the orientation of the hands revealed that these congruency effects were determined by the position of the target and distractor in external space, and not by the particular skin sites stimulated (Experiment 3). Congruency effects increased as the hands were brought closer together in the dark (Experiment 4), demonstrating the role of proprioceptive input in modulating tactile selective attention. This spatial modulation was also demonstrated when a mirror was used to alter the visually perceived separation between the hands (Experiment 5). These results suggest that tactile, spatially selective attention can operate according to an abstract spatial frame of reference, which is significantly modulated by multisensory contributions from both proprioception and vision. PMID- 15751468 TI - The relationship between perceived length and egocentric location in Muller-Lyer figures with one versus two chevrons. AB - We examined the apparent dissociation of perceived length and perceived position with respect to the Muller-Lyer (M-L) illusion. With the traditional (two chevron) figure, participants made accurate open-loop pointing responses at the endpoints of the shaft, despite the presence of a strong length illusion. This apparently non-Euclidean outcome replicated that of Mack, Heuer, Villardi, and Chambers (1985) and Gillam and Chambers (1985) and contradicts any theory of the M-L illusion in which mislocalization of shaft endpoints plays a role. However, when one of the chevrons was removed, a constant pointing error occurred in the predicted direction, as well as a strong length illusion. Thus, with one-chevron stimuli, perceived length and location were no longer completely dissociated. We speculated that the presence of two opposing chevrons suppresses the mislocalizing effects of a single chevron, especially for figures with relatively short shafts. PMID- 15751470 TI - The effect of vertical tongue loading on the position perception of the tongue. AB - We investigated the effects of vertical tongue loading on the position perception of the tongue. Five male and 5 female university students served as subjects. Vertical upward and downward loading forces were applied to the tongue of the subjects. Their task was to judge the perceived horizontal position of the tongue after tongue-loading directions. The means of the judgments for the control conditions (no tongue loading) were compared with the judgments for perceived horizontal position after tongue loading. The results showed that vertical tongue loading produced a shift in the perceived horizontal direction opposite to the applied force. These results fully replicated the analogous aftereffect found by Grover and Craske (1991) for horizontal tongue loading. However, the judgments of perceived horizontal position in the present study had lower variability than did those in Grover and Craske's study, suggesting that mapping of the tongue along the vertical axis is more precise than mapping along the horizontal axis. PMID- 15751469 TI - Judgments of duration, figure-ground contrast, and size for words and nonwords. AB - Does the word-superiority effect on letter discrimination result in a word superiority effect on duration judgments? We examined this question in five experiments. In the first four experiments, we have demonstrated that (1) words shown for 32-80 msec were judged as presented longer than non-words shown for the same duration; (2) this word-superiority effect persists if the stimuli are shown for an objective duration of up to 250 msec; and (3) these effects can be extended to judgments of figure-ground contrast and letter size. These findings extend existing data on effects of processing fluency on perceptual judgments. In Experiment 5, we found that duration judgments were higher for words than for pronounceable nonwords, and duration judgments were higher for pronounceable non words than for nonpronounceable nonwords. We discuss the implications of this finding for the discrepancy-attribution hypothesis. PMID- 15751472 TI - Revisiting within-modality and cross-modality attentional blinks: effects of target-distractor similarity. AB - When two masked targets (T1 and T2) require attention and are presented within half a second of each other, the report accuracy for T2 is reduced, relative to when the two targets are presented farther apart in time. This effect is known as the attentional blink (AB). Potter, Chun, Banks, and Muckenhoupt (1998) argued that all AB-like effects observed when at least one of the targets was presented outside of the visual modality did not represent true instances of the AB, but instead were artifacts of task-set switching. However, in the Potter et al. experiments the presence or absence of task-set switching opportunities was confounded with the T2 task, as well as the alphanumeric class of T2 with respect to the distractors. In the present experiment, we examine the influence of T1 alphanumeric class, T2 alphanumeric class, and switching operations in a fully crossed design that unconfounds these factors. In contrast to the conclusions of Potter et al., the present results suggest that the T2 alphanumeric class can account for the pattern of ABs observed across conditions, without necessarily implicating a separate switch cost. The implications for theoretical models of the AB and the debate over the validity of cross-modal ABs are discussed. PMID- 15751471 TI - Modeling panel detection frequencies by queuing system theory: an application in gas chromatography olfactometry. AB - In continuous vigilance tasks, the number of coincident panel responses to stimuli provides an index of stimulus detectability. To determine whether this number is due to chance, panel noise levels have been approximated by the maximum coincidence level obtained in stimulus-free conditions. This study proposes an alternative method by which to assess noise levels, derived from queuing system theory (QST). Instead of critical coincidence levels, QST modeling estimates the duration of coinciding responses in the absence of stimuli. The proposed method has the advantage over previous approaches that it yields more reliable noise estimates and allows for statistical testing. The method was applied in an olfactory detection experiment using 16 panelists in stimulus-present and stimulus-free conditions. We propose that QST may be used as an alternative to signal detection theory for analyzing data from continuous vigilance tasks. PMID- 15751473 TI - The locus of perceived equidistance in binocular vision. AB - Empirical studies of the locus of perceived equidistance in binocular vision have revealed a characteristic change of its form, depending on absolute distance. This result is commonly taken to indicate influence of vergence-related binocular information, a conclusion that is by no means exclusively dictated by the data. Heller (1997) has suggested an alternative theoretical account that is based on the idea of independently combining the outcome of monocular input transformations without any form of binocular interaction. This article provides an experimental test of the structural assumption lying at the core of the axiomatic foundation of Heller's theory. I test the so-called Reidemeister condition under reduced cue conditions in two settings for each of 7 subjects. The results provide strong evidence for the validity of the Reidemeister condition and thus challenge the view that the locus of perceived equidistance depends on vergence-related binocular information. The discussion of the factors contributing to the monocular input transformations emphasizes the role of the optical properties of the eyes. PMID- 15751474 TI - Attention and the subjective expansion of time. AB - During brief, dangerous events, such as car accidents and robberies, many people report that events seem to pass in slow motion, as if time had slowed down. We have measured a similar, although less dramatic, effect in response to unexpected, nonthreatening events. We attribute the subjective expansion of time to the engagement of attention and its influence on the amount of perceptual information processed. We term the effect time's subjective expansion (TSE) and examine here the objective temporal dynamics of these distortions. When a series of stimuli are shown in succession, the low-probability oddball stimulus in the series tends to last subjectively longer than the high-probability stimulus even when they last the same objective duration. In particular, (1) there is a latency of at least 120 msec between stimulus onset and the onset of TSE, which may be preceded by subjective temporal contraction; (2) there is a peak in TSE at which subjective time is particularly distorted at a latency of 225 msec after stimulus onset; and (3) the temporal dynamics of TSE are approximately the same in the visual and the auditory domains. Two control experiments (in which the methods of magnitude estimation and stimulus reproduction were used) replicated the temporal dynamics of TSE revealed by the method of constant stimuli, although the initial peak was not apparent with these methods. In addition, a third, control experiment (in which the method of single stimuli was used) showed that TSE in the visual domain can occur because of semantic novelty, rather than image novelty per se. Overall, the results support the view that attentional orienting underlies distortions in perceived duration. PMID- 15751475 TI - Detecting malingerers by means of response-sequence analysis. AB - The feigning of sensory loss (malingering) poses a challenge for psychophysicists. To uncover malingerers by means of psychophysical testing, we combined measures of response-sequence randomness with the maximum-likelihood adaptive-staircase procedure used to measure sensory detection thresholds. The two-alternative, forced-choice maximum-likelihood adaptive-staircase procedure calculates an estimate of the threshold after each trial and also recommends the stimulus concentration for the next trial. Olfactory detection thresholds for butyl alcohol were measured in 7 normals, 6 anosmics, and 6 malingerers. Each participant was tested for 20 trials. A discriminant analysis, using threshold concentration and probability of being correct over the 20 trials, could correctly classify only 68% of the malingerers and anosmics. Correct classification of anosmics and malingerers rose to 100% when statistical measures of randomness in the response sequences were included in the discriminant analysis. We conclude that the maximum-likelihood adaptive-staircase procedure, combined with response-sequence analysis, is a powerful addition to the arsenal of techniques for detecting malingerers in the evaluation of sensory ability. PMID- 15751476 TI - On the use of discrimination to assess memory. AB - A number of studies have made use of sensory discrimination to assess memory. These experiments have typically employed a temporal two-alternative forced choice paradigm in which the interstimulus interval (ISI) is varied. Many of these tests have shown that ISI has little or no effect on discrimination performance. This has been taken as evidence for perfect or very good memory. An alternative interpretation would be that discrimination tests lack sufficient sensitivity to reveal memory effects. In the present study, computer simulations based on signal detection theory were used to estimate the relative effects of memory noise and direct noise (i.e., noise inherent in the discrimination task) on discrimination performance. These simulations indicate that the amount of memory noise has to be quite large to cause a marked elevation in discrimination thresholds and that sensory discrimination is a relatively insensitive test of memory. PMID- 15751477 TI - Choice and response time processes in the identification and categorization of unidimensional stimuli. AB - Lacouture and Marley (1991, 1995, 2001) have successfully modeled the probabilities of correct responses and the mean correct response times (RTs) in unidimensional absolute identification tasks for various stimulus ranges and stimulus/response set sizes, for individual and group data. These fits include those to a set of phenomena often referred to as end-anchor effects. A revised model, with the independent accumulator decision process replaced by a leaky competing accumulator decision process, fits the probabilities of correct responses and the full distributions of RTs in unidimensional absolute identification. The revised model is also applied successfully to a particular class of unidimensional categorization tasks. We discuss possible extensions for handling sequential effects in unidimensional absolute identification, and other extensions of the given class of categorization tasks that are of potential empirical and theoretical importance as a supplement to the study of multidimensional absolute identification tasks. PMID- 15751478 TI - An eye movement analysis of "mental rotation" of simple scenes. AB - Participants saw a standard scene of three objects on a desktop and then judged whether a comparison scene was either the same, except for the viewpoint of the scene, or different, when one or more of the objects either exchanged places or were rotated around their center. As in Nakatani, Pollatsek, and Johnson (2002), judgment times were longer when the rotation angles of the comparison scene increased, and the size of the rotation effect varied for different axes and was larger for same judgments than for different judgments. A second experiment, which included trials without the desktop, indicated that removing the desktop frame of reference mainly affected the y-axis rotation conditions (the axis going vertically through the desktop plane). In addition, eye movement analyses indicated that the process was far more than a simple analogue rotation of the standard scene. The total response latency was divided into three components: the initial eye movement latency, the first-pass time, and the second-pass time. The only indication of a rotation effect in the time to execute the first two components was for z-axis (plane of sight) rotations. Thus, for x- and y-axis rotations, rotation effects occurred only in the probability of there being a second pass and the time to execute it. The data are inconsistent either with an initial rotation of the memory representation of the standard scene to the orientation of the comparison scene or with a holistic alignment of the comparison scene prior to comparing it with the memory representation of the standard scene. Indeed, the eye movement analysis suggests that little of the increased response time for rotated comparison scenes is due to something like a time-consuming analogue process but is, instead, due to more comparisons on individual objects being made (possibly more double checking). PMID- 15751479 TI - Evidence for patchwork approximation of shape primitives. AB - Investigators have proposed that qualitative shapes are the primitive information of spatial vision: They preserve an approximately one-to-one mapping between surfaces, images, and perception. Given their importance, we examined how the visual system recovers these primitives from sparse disparity fields that do not provide sufficient information for their recovery. We hypothesized that the visual system interpolates sparse disparities with planes, resulting in a patchwork approximation of the implicitly defined shapes. We presented observers with stereo displays simulating planar or smooth curved surfaces having different curvatures. The observers' task was to detect whether dots deviated from these surfaces or to discriminate planar from curved or planar from scrambled surfaces. Consistent with our hypothesis, increasing curvature had detrimental effects on observers' performance (Experiments 1-3). Importantly, this patchwork approximation leads to the recovery of the proposed shape primitives, since observers were more accurate at discriminating planar-from-curved than planar from-scrambled surfaces with matched disparity range (Experiment 4). PMID- 15751480 TI - Maximizing potential: federal financing for treatment of adolescents with substance use disorders. AB - The federal government is making a significant contribution to the treatment of adolescents with substance use disorders. However, funds are scattered across a number of policy domains. While some federal policies are complementary, many overlap, illustrating not only the complexity of collaborative efforts on the part of the implementing federal agencies but also the diversity of partnerships supporting adolescent substance use treatment. Changes in the organization and financing of health care delivery, coupled with the complex interaction of federal policies that address this population, affect the efficiency and effectiveness of federal policy responses to the problem. Equally important are the gaps in federal policy and the inadequacy of federal programs to reach all youth in need. This article analyses existing federal policies and programs and concludes that a seamless, multidisciplinary system that transcends traditional federal policy boundaries must be designed and implemented. PMID- 15751481 TI - Systems of care for treatment of adolescent substance use disorders: background, principles and opportunities. AB - Adolescent substance abuse is a serious social problem facing the United States. Despite numerous recent advances in the clinical effectiveness of treatment approaches for this population, not enough attention has been paid to the adolescent treatment and service delivery infrastructures. The right services must be delivered through carefully organized and systematic community partnerships among agencies that serve the youth and families most in need. This article provides a working definition for the systems of care approach, reviews the movement's history within children's mental health services, addresses the feasibility of using the systems of care model for adolescent substance use disorders, and discusses principles and elements essential for successfully implementing a system of care for treatment of adolescent substance use disorders. PMID- 15751482 TI - Pathways to chemical dependency treatment for adolescents in an HMO. AB - This study examines pathways to chemical dependency (CD) treatment for adolescents in a prepaid health plan, including factors influencing treatment referral and access. Data were collected from 419 adolescent CD treatment intakes ages 13 to 18 in a large, private health plan. The study examines predictors of referral from different sources and of mental health treatment prior to CD treatment. Referral patterns, problem severity, and psychiatric comorbidity differed by gender. Being male (p < .05) and higher scores on the Youth Self Report (YSR) externalizing scale (p < .01) predicted a justice system referral. Using more types of substances predicted referral by medical and mental health providers (p < .05) and self-referral (p < .01). Higher YSR internalizing scores predicted referral from mental health (p < .01) and self-referral (p <.01). Being White versus African American (p < .05) or Latino (p < .01), older (p < .05), and having higher YSR internalizing scores (p < .05), a conduct disorder (p < .01), or a family member with a substance use problem (p < .01) predicted a mental health visit prior to CD intake. The findings raise questions regarding the role of health plans, clinicians, families, schools, and community agencies in referring and treating adolescents with substance use problems. They suggest that improved coordination of care may promote more integrated treatment practices, which could decrease substance use, mental health, and medical problem severity. PMID- 15751483 TI - Co-occurring disorders in the adolescent mental health and substance abuse treatment systems. AB - This article explores the rates of co-occurring disorders in two large federally funded programs that target youth. In the mental health treatment system, the Substance Abuse and Mental Health Services Administration's (SAMHSA) Center for Mental Health Services (CMHS) supports the Comprehensive Community Mental Health Services for Children and Their Families Program. SAMHSA's Center for Substance Abuse Treatment (CSAT) supports a number of grant programs providing substance abuse treatment for adolescents. The data from these programs underscores the need for the use of systematic, validated, biopsychosocial assessment instruments for all youth entering either the substance abuse or mental health treatment systems. The current evidence base for models of co-occurring treatment for youth is discussed and recommendations made for future activity related to adolescent co-occurring treatment. PMID- 15751485 TI - Using administrative data to assess the process of treatment services for adolescents with substance use disorders. AB - In 1998, the Center for Substance Abuse Treatment (CSAT), Substance Abuse and Mental Health Services Administration (SAMHSA) convened an expert panel comprised of a multidisciplinary group of providers, researchers, managed care representatives and public policymakers known as the Washington Circle (WC) to improve the quality and effectiveness of treatment for substance use disorders through the use of performance measures. The initial goals of the panel included developing, testing and implementing performance measures in health plans and encouraging collaboration with key stakeholders to ensure widespread adoption of the measures. The WC focused its initial efforts on the prevention, recognition and treatment of adults with alcohol and other drug (AOD) disorders who receive health care through private and/or public sector health plans (McCorry et al. 2000). In 2002, CSAT, aware of the need to improve access and quality of treatment for adolescents, supported the Washington Circle in the establishment of a special subcommittee focused specifically on adolescent treatment issues. This article introduces the work of the Washington Circle and describes the Adolescent Subcommittee's progress in developing performance measures for the treatment of adolescents with substance use disorders. PMID- 15751484 TI - Effects of alcohol and drug use on inpatient and residential treatment among youth with severe emotional disturbance in Medicaid-funded behavioral health care plans. AB - This study examined the use of alcohol and recreational drugs among 875 youth with severe emotional disturbance (SED) enrolled in Medicaid-funded behavioral health care plans, and whether co-occurring SED and substance use affected the subsequent likelihood of receiving inpatient and/or residential treatment. Youth at five sites nationwide were interviewed about their use of drugs and alcohol, while interviews with their caregivers elicited information about youths' service utilization, degree of functional impairment, and a series of demographic and environmental variables. Results indicated that half of the youth (52%) reported lifetime use of alcohol, street drugs, or over-the-counter medications for recreational purposes, while 18% reported use in the past 30 days. Among those reporting recent use, 32% reported using drugs only, 34% alcohol only, and 33% reported use of both drugs and alcohol. In multivariate logistic regression analyses, the effect of recent use was stronger than that of lifetime use; however, the largest effect occurred for those reporting recent use of both drugs and alcohol, versus either alone, or none. Differences remained significant when controlling for managed care versus fee for service enrollment as well as child, family, and environmental characteristics including study site. These results mirror those of prior studies that found an association between substance use and greater likelihood of inpatient services, even in managed care settings. PMID- 15751486 TI - The adolescent addiction treatment workforce: status, challenges, and strategies to address their particular needs. AB - Treating youth with substance use problems presents unique challenges, distinct from those of adults, that add to the complexity of recruiting and retaining a qualified adolescent treatment workforce. Youth who need treatment are often involved in an array of systems (i.e., health, legal, and social) and use substances for a variety of reasons, requiring providers to have a unique set of treatment practices and proficiencies. The intent of this article is to highlight the current understanding of the issues facing the alcohol and other drug treatment workforce and to provide an overview of strategies people are or could be using to build and maintain a workforce that can effectively deliver care for adolescents. PMID- 15751487 TI - Snapshot of the substance abuse treatment workforce in 2002: a synthesis of current literature. AB - Substance abuse among adolescents and adults continues to be a major public health concern. Given the prevalence of substance use, abuse, and dependence in the United States, the treatment needs of the population who abuse substances are great. Adolescents and adults who abuse substances need competent, knowledgeable, and qualified staff to provide services to meet their treatment needs. However, providers of substance abuse treatment services are varied, ranging from those who have minimal formal training to those who have specialized degrees and credentials in the field. In addition, substance abuse professionals represent a variety of fields (social work, psychiatry, psychology, etc) as opposed to a single unifying discipline. Few studies have been conducted examining the background, qualifications, and professional development needs of treatment staff. This article represents an attempt to lay the groundwork for future research. It summarizes information on staff demographics, level of competency, training, recruitment, and retention. In addition, recommendations are made for the advancement of research. PMID- 15751488 TI - Certification and program regulations for inpatient services to youth with addiction: a state-level analysis. AB - Recent evidence has indicated that the prevalence of addiction in youth is far higher than previously expected. High demand for services to this population require attention to programs providing interventions, in particular to the regulations and structures that assure youth receive quality services. Thus, the current research examines state-level certification and program requirements for inpatient youth facilities. Current data (from August 2002 through July 2003) on state-level regulations was collected from a variety of sources. Wherever possible, confirmation was sought from multiple sources. Multiple authors interpreted information. Youth with addictions are currently being treated across a wide variety of governmental agencies. Level of detail and clarity of information varied widely, as did distinctness of youth requirements from adult ones. Adequate regulatory attention was evident in a minority of states, while a handful of states did little or nothing to distinguish youth from adult services. While it is undoubtedly the case that providers within each state offer outstanding services, there is often little or no available protection for these youth through governmental regulations. Attention is needed in almost all states to providing additional regulations and program requirements that assure youth consistency in service quality. Recommendations are made for standards of certification requirements. PMID- 15751489 TI - Dental pain in Maryland school children. AB - OBJECTIVE: To describe the lifetime prevalence of dental pain among Maryland's school age children. METHODS: Data come from the Survey of Oral Health Status of Maryland School Children conducted in 2000-01. History of dental pain, as reported by an adult respondent, was analyzed for 2,411 kindergarten and third grade students. Percentages with their 95% confidence interval were calculated to obtain state representation of the distribution of dental pain by sociodemographic characteristics and caries status. RESULTS: Overall, 11.8 percent of Maryland school age children in kindergarten and third grade have had some dental pain. Among children who have had caries, the report of dental pain increases to 28.2 percent. Children from families with low educational attainment or eligible for free or reduced meals or covered by Medicaid were more likely to have experienced dental pain. CONCLUSION: Almost a third of Maryland kindergartens and third graders who have caries have experienced dental pain. PMID- 15751490 TI - Patterns of dietary fluoride supplement use in children from birth to 96 months of age. AB - OBJECTIVE: This paper reports on fluoride supplement use from birth to age 96 months. METHODS: As part of the Iowa Fluoride Study, questionnaires were mailed at 3 to 6- month intervals assessing use of supplements. Estimated fluoride intake from supplements was calculated as a daily fluoride intake in mg. Analyses include descriptive statistics of supplements use for individual periods, area under-the-curve (AUC) for combined periods, and associations between fluoride supplement use and demographic covariates. RESULTS: Participants (n=1,388) were mostly white, with about two-thirds of parents having some college education. Percentages using fluoride supplements were 11.2% (12 months), 6.3% to 6.8% (24, 36, 48 and 60 months) and 3.6% to 4.7% (72, 84 and 96 months). Physicians prescribed most supplements until age three. The mean supplement dosage when used gradually increased from 0.25 mg (12 months) to 0.82 mg (84 months) and 0.75 mg (96 months). The effective mean daily fluoride supplement quantities ingested among users for the successive age groups from 12 to 96 months old were 0.14, 0.14, 0.25, 0.34, 0.37, 0.43, 0.48, and 0.37 mg, respectively. Estimated daily average fluoride ingested from supplements increased from 0.06 mg (birth-12 months) to 0.07 mg (12-60 months) to 0.18 mg (60-96 months). CONCLUSION: Fluoride supplement use patterns varied substantially among individuals; however, average use within the intervals birth-12 months, 12-60 months, and 60-96 months was fairly consistent. PMID- 15751491 TI - Associations between socio-economic circumstances at two stages of life and adolescents' oral health status. AB - There is a consistent association between unfavourable socio-economic circumstances and oral health. Although the effects of poor social circumstances in childhood are known to have lasting influences on general health, there is little information on their effects regarding chronic oral diseases. OBJECTIVE: To assess the relationship between oral health status and socio-economic circumstances at two different periods of adolescents' life. METHODS: A two-phase cross sectional study was carried out in Brazil. In Phase I, 652 13-year-olds were clinically examined and interviewed. In the second phase, 311 families were randomly selected for in-depth interviews. Information was collected on several indicators of socio-economic circumstances, family related variables, school grade level, and oral health behaviour, at two different life stages, at birth and at 13 years of age. The outcome variable was oral health status at the age of 13. It was constructed by counting the worst scores of DMFT, gingival bleeding, calculus and dental plaque. The data analysis used stepwise logistic regression. RESULTS: The response rates for phases I and II were 85% and 94%. Boys, those at a lower grade level at school for their age, and those who experienced high levels of material deprivation at birth and at the age of 13 were more likely to have high levels of oral diseases; the odds ratios were 4.12 (1.86-9.16), 2.41 (1.01-5.76) and 4.61 (1.30-16.3), respectively. CONCLUSION: Brazilian adolescents experiencing adverse socio-economic circumstances at birth and at the age of 13 had high levels of oral diseases. PMID- 15751492 TI - The use of administrative databases to assess oral health care. AB - OBJECTIVE: We examined the potential for research using administrative databases containing dentists' claims to identify both the type of health services research questions addressed and the strength of the evidence that is achieved in such studies. METHODS: We searched Medline (1966 to March, 2003), retrieved additional reports from personal files, reviewed the literature cited in the relevant articles and conducted electronic searches on investigators' surnames. Information from relevant articles was abstracted into tables and the strength of the evidence for each was classified. RESULTS: Thirty-eight studies met our inclusion criteria. Researchers have used administrative databases of dental records to examine provider practices, the longevity or consequences of dental interventions, the prevalence of dental conditions, and patient factors that determined care, and to establish quality assurance criteria or standards of care. The strongest designs were prospective or case-control (Level II-2). CONCLUSION: Studies analyzing administrative databases have the advantage of size and economy but are subject to several threats to their validity and are seldom population-based. The strongest designs occurred with investigation of the longevity or consequences of care. Several studies demonstrated the benefit of linking the service data to patient or provider characteristics. The study of dentists' claims data appears under exploited, especially in the area of identifying and recommending changes in dental health care policies. PMID- 15751493 TI - Healthy practices, social relationships and dental perceptions among US dentate adults. AB - OBJECTIVE: This study explored and described the effects of healthy practices and social relationships on dental perceptions among U.S. dentate adults. METHODS: Guided by the Alameda County Study: Health and Ways of Living, weighted data from the Third National Health and Nutrition Examination Survey of 8,449 U.S. dentate adults aged 18-64 years were analyzed with SUDAAN 7.5.3. RESULTS: Healthy practices and social relationships showed protective effects against unfavorable dental perceptions after controlling for socio-demographic characteristics, dental access, and related clinical dental status. Besides healthy practices and social relationships, education, family income, having dental visit in the past 12 months, having dental decay, having defective tooth condition, and number of natural teeth helped explain unfavorable dental perceptions in the adjusted model. Compared with persons who had high social relationships and high healthy practices, those who had low social relationships with very low, low and medium healthy practices were about five, times (OR=5.07; 99% CI=3.24, 7.91), almost four times (OR=3.61; 99% CI=2.22, 5.87), and two times (OR=2.24; 99% CI=1.37, 3.67) more likely, respectively, to have unfavorable dental perceptions. In the adjusted model, these odds ratios decreased to three (OR=3.30; 99% CI=2.01, 5.41), two (OR=2.06; 99% CI=1.15, 3.69) and non-significant difference, consecutively. CONCLUSIONS: Influence of healthy practices and social relationships on dental health may encourage dental health professions to participate in general health behavior modifications and social actions to foster social relationships, in addition to preventive dental care. PMID- 15751494 TI - Knowledge, attitude and practice towards droplet and airborne isolation precautions among dental health care professionals in Shiraz, Iran. AB - OBJECTIVE: To determine the level of knowledge, attitude and practice among Iranian dental health care professionals towards droplet and airborne isolation precautions, in Shiraz, Iran. METHODS: A survey was developed and distributed to 152 Iranian dental professionals (faculty and students) from Shiraz University School of Medical Sciences, School of Dentistry, Shiraz, Iran. The self administered questionnaire was composed of three parts (knowledge, attitude, and practice). After validation of the survey using the Kuder Richardson reliability test and Cronbach's alpha internal consistency coefficient, data was collected and analyzed using Analysis of Variance and the Duncan test to detect the differences among groups. The Spearman coefficient was used to determine the correlation between knowledge, attitude, and practice. RESULTS: The survey was completed by 152 (51.5%) of 295 potential respondents. Mean scores of knowledge, attitude, and practice towards droplets and airborne isolation precautions were poor (98 +/- 3.45, 10.51 +/- 6.26, 2.68 +/- 3.16 from the maximum scores of 11, 55, and 11, respectively). In addition, a positive linear correlation was found between two items of the survey including knowledge-attitude (rho = 0.438, p < 0.001), knowledge and practice (rho = 0.380, p < 0.001). CONCLUSION: Iranian dental professionals seemed to have low levels of knowledge, attitude and practice towards infection control. PMID- 15751495 TI - Developing and deploying a new member of the dental team: a pediatric oral health therapist. AB - There are inadequate numbers of dentists able and willing to treat America's children, specifically children from low income and minority populations. This has led to the well-publicized disparities in oral health among children. In the early part of the 20th century New Zealand faced a significant problem with oral disease among its children and introduced a School Dental Service, staffed by allied dental professionals with two years' training in caring for the teeth of children, "school dental nurses." A significant number of countries have adopted the model. This article reviews the history of attempts to develop such an approach in the United States. It advocates for the development and deployment of pediatric oral health therapists as a means of addressing the disparities problem that exists in America with such individuals being trained in children's dentistry in a two-year academic program. The article asserts that adding a pediatric oral health therapist to the dental team is one way in which the profession of dentistry can fulfill its moral obligation to care for the oral health of America's children and ensure that all children are treated justly. Recently, the American Association of Public Health Dentistry promulgated a strategic plan that endorsed such an approach. PMID- 15751496 TI - Public health implications of periodontal infections in adults: conference proceedings. PMID- 15751497 TI - Therapeutic options in the management of colon cancer: 2005 update. AB - Recently, major developments in the treatment of colon cancer have emerged. These developments include improvements in surgical technique and staging and the introduction of new molecularly targeted pharmacologic agents. Improvements in surgical management involve enhanced staging techniques, allowing more accurate determination of risk of recurrence. Newer agents, such as oxaliplatin, cetuximab, and bevacizumab, now are approved for the treatment of colon cancer. The data associated with use of oxaliplatin in adjuvant and metastatic settings continue to mature; survival benefits are expected to become more fully apparent in the next two years. Bevacizumab, a monoclonal antibody that neutralizes vascular endothelial growth factor, when combined with irinotecan, 5 fluorouracil, and leucovorin (IFL), was superior to IFL alone in achieving median and progression-free survival. Cetuximab, a monoclonal antibody directed against the epidermal growth factor receptor, when given in combination with irinotecan, achieved an increased objective response and increased time to progression, compared with cetuximab alone, in patients refractory to irinotecan-containing regimens. In addition to surgical and pharmacologic developments, the recognition that genetics and molecular markers play an important role in carcinogenesis has heightened research to integrate this knowledge into practice. Nurses play a pivotal role in the care of patients with colon cancer and must be conversant in the new advances in treatment. PMID- 15751498 TI - Outpatient genetic risk assessment in women with breast cancer: one center's experience. AB - A chart audit at one cancer center, of 193 women with breast cancer, was completed to assess whether a complete family history that may indicate genetic predisposition was obtained and if that information led a provider to suggest risk reduction strategies. A risk management tool, which included a pedigree template, was used. Of the 193 charts reviewed, 88.6% had family history information recorded; 41.5% reported three generations of family history. Risk management was undocumented in 21.8% of the charts reviewed and, for those that were reported (78.2%), 7.25% were referred for genetic counseling. These results suggest that a more detailed assessment of hereditary breast cancer risk incorporating three generations of family history and additional types of cancer need to be integrated into medical oncology practice. An algorithm was developed as a guide to improve the process of evaluation and referral for genetic risk assessment. PMID- 15751499 TI - Antivascular endothelial growth factor monoclonal antibody therapy: a promising paradigm in colorectal cancer. AB - Angiogenesis plays an important role in tumor growth and development. Vascular endothelial growth factor (VEGF) is one of the most potent proangiogenic factors and therefore is an ideal target in colorectal cancer therapy. Bevacizumab (Avastin, Genentech, Inc., South San Francisco, CA) is a humanized monoclonal antibody, designed to directly target VEGF. The agent has shown promising activity in preclinical and phase I and II studies and is well tolerated compared with conventional cytotoxic chemotherapy. The U.S. Food and Drug Administration recently approved bevacizumab in combination with 5-fluorouracil-based chemotherapy as first-line therapy for patients with metastatic colorectal cancer. The approval was based on phase III data demonstrating that patients treated with bevacizumab plus chemotherapy survived approximately five months longer compared with patients treated with chemotherapy alone. This article will focus on the role of VEGF in tumorigenesis and summarize the available data on the use of bevacizumab in the treatment of metastatic colorectal cancer. PMID- 15751500 TI - Management of patients who have undergone hepatic artery chemoembolization. AB - Hepatocellular carcinoma (HCC) is the fifth most common cause of all malignancies and causes approximately one million deaths each year. Surgical liver resection is the only cure for HCC; however, few patients are eligible to undergo this procedure. Hepatic artery chemoembolization (HACE) is a technique that delivers high concentrations of chemotherapeutic agents and blocks the blood supply to the liver for prolonged periods of time. HACE has demonstrated an overall increase in survival. The HACE procedure, pre- and postprocedure complications, and the care required by patients with HCC are critical for oncology nurses to understand. PMID- 15751501 TI - Integrating aprepitant and palonosetron into clinical practice: a role for the new antiemetics. AB - Chemotherapy-induced nausea and vomiting (CINV) are among the most feared side effects of cancer treatment. With increasingly more complex chemotherapy treatments, CINV plays an important role in determining patients' quality of life, as well as when to halt potentially lifesaving therapy. Although significant progress has been made in the treatment of CINV, patients undergoing chemotherapy continue to report that this side effect is persistent and distressing. In 2003, two new agents were added to the armamentarium of antiemetic therapy. The U.S. Food and Drug Administration approved palonosetron, a longer-acting serotonin antagonist, and aprepitant, a neurokinin-1 antagonist and the first in a new class of antiemetics, for the treatment of CINV. Although the indications for both agents are similar, they have distinct differences. Decisions regarding placement of these agents into existing antiemetic protocols can be based on national guidelines, review of the literature, and clinical experience. This article will review current antiemetic therapy with an emphasis on the new additions to the treatment of CINV. Aprepitant and palonosetron represent significant changes in the treatment of CINV. Oncology nurses need to know current approaches to maximize effective antiemetic therapy. PMID- 15751502 TI - Patient-specific vaccine therapy for non-Hodgkin lymphoma. AB - Follicular non-Hodgkin lymphoma (NHL) is an indolent, or slow-growing, malignant disease of the lymphoid tissue, which usually responds to initial therapy. However, the disease is characterized by multiple relapses and remissions, eventually causing death. Several effective therapies are available, but improvement of overall survival in patients with follicular NHL has not been demonstrated. Stimulation of the immune system to recognize malignant lymphoma cells as foreign has been demonstrated as a viable treatment option for patients with follicular NHL. Patient-specific vaccine therapy is a new form of active immunotherapy being studied for NHL. Clinical trials have shown a benefit for patients receiving this type of therapy. This article will provide a foundation for nurses caring for patients receiving patient-specific vaccine therapy. PMID- 15751503 TI - A day in the life of Suzanne M. Mahon, cancer prevention expert. PMID- 15751504 TI - Palmar-plantar erythrodysesthesia. PMID- 15751505 TI - Oral chemotherapy. PMID- 15751506 TI - Reading a research article part I: Types of variables. PMID- 15751507 TI - Complementary and alternative medicine patients are talking about: PC-SPES. PMID- 15751509 TI - [Development of gastrotonometry and the future of its use in the monitoring of critically ill patients]. AB - Gastrotonometry conventionally enables the measurement of the partial pressure of carbon-dioxide (pCO2) in the gastric mucosa via the analysis of the filling material of a ballooned probe placed into the stomach. According to the data of the more than a half century long history of the method we may state, that the examination triggered an unusually large interest as well as was applied both in clinical work and in research. Since in the case of good circulatory circumstances the result of the examination is nearly equal to the value of the arterial blood pCO2, in the beginning the method was used for the better setting of ventilators of poliomyelitic patients with respiratory paralysis. Later it was also explored, that in critically ill patients the pCO2 value is significantly higher, than that of the arterial blood value, which difference is the consequence of the disturbance in the regional perfusion of the gastrointestinal system. The attention toward the subject was mainly drawn by the circumstance that the examination of the patient's condition can be judged objectively. Even more important is that gastrotonometry may help in the prediction of a life threatening event. One of the main purposes of the article was the summarising of the literature on the subject, as well as reviewing the aspects, which should urge clinicians to apply gastrotonometry in a broader range of everyday clinical practice. On the other hand, the authors also give a short description of a new, self-developed balloon-free probe. PMID- 15751508 TI - [Achieving the target lipid levels in Hungary, 2004]. AB - INTRODUCTION: Recent European and Hungarian guidelines of cardiovascular prevention have clearly defined the target levels in lipid lowering therapy. OBJECTIVE: To analyze the risk status of patients receiving long term lipid lowering therapy and the rate of achievement of target level in a Hungarian multicenter trial. METHOD: The investigation was performed in January and February of 2004 involving general practitioners and specialists (cardiologists, lipidologists). Applying a questionnaire the authors asked for risk factors and the further implementations in the knowledge of results of each doctors' 10 consecutive patients receiving lipid lowering therapy for at least one year. RESULTS: LDL target levels accepted by the Hungarian Therapic Consensus Conference was achieved in 22% of GP's patients and 27% of specialists' patients, in 24% of patients, in average. According to risk stratification, the 83% of patients receiving lipid lowering therapy were at high risk, and 79% of these did not reach the suggested target level of serum total cholesterol. In 54% of patients not achieving target level the doctors continue the therapy without any modification. Considering therapy modification in the rest of patients, a dose increase was intended in 61%, change of drug in 31% of cases, and rarely the combination therapy. CONCLUSION: While only a quarter of patients receiving lipid lowering treatment achieved the target levels, along with wide spreading of this kind of therapy further efforts should be made in order to achieve the levels defined in guidelines with dose increase or combination of appropriate drugs. PMID- 15751510 TI - [Preliminary experience with controlled lumbar drainage in medically refractory intracranial hypertension]. AB - INTRODUCTION: The contribution of brain edema to brain swelling in cases of traumatic brain injury remains a critical problem. In head injury, the swelling and eventual rise in intracranial pressure is a frequent cause of death, and in survivors the poor prognosis with sustained elevation of ICP has been well documented. OBJECTIVE: The objective this study was to evaluate the effect of controlled lumbar cerebrospinal fluid drainage in adult patients with refractory intracranial hypertension following severe brain injury. METHOD: The study involved 10 head injured patients (GCS < or = 8) with medically refractory intracranial hypertension. Aggressive treatment included the repeated steps of the Brain Trauma Foundation's guidelines, barbiturate coma and in many cases decompressive craniectomy as well. After institution of a lumbar drain, cerebrospinal fluid drainage was maintained under control of intracranial pressure (ICP) and neurological status. ICP and cerebral perfusion pressure before and after initiation of lumbar cerebrospinal fluid drainage and related complications were documented. RESULTS: All patients demonstrated an immediate decrease of ICP (from 30.6 +/- 4.7 mm Hg to 11.5 +/- 3.9 mm Hg, mean +/- SD) and a concomitant increase of cerebral perfusion pressure. In seven patients the decrease of ICP was long lasting and 5 of them had a favourable outcome. Two patients survived with a severe permanent neurologic deficit and only three patients died because of the progressive brain edema, which developed despite of the maximum therapy. CONCLUSION: In conclusion we may consider, that controlled lumbar cerebrospinal fluid drainage is a potentially useful treatment in cases of severe traumatic brain injury when maximal medical therapy and ventricular cerebrospinal fluid evacuation have failed to control high intracranial hypertension. The danger of herniation is minimized by considering lumbar drainage in the presence of discernible basilar cisterns only. PMID- 15751512 TI - [Treatment of primary glomerulonephritis]. PMID- 15751511 TI - [Soluble syndecan-1 levels in different plasma cell dyscrasias]. AB - INTRODUCTION: Syndecans are a family of cell surface proteoglycans. In the bone marrow of multiple myeloma patients syndecan-1 is expressed only on the surface of malignant plasma cells. The aim of the study was to determine the soluble syndecan-1 levels in different plasma cell dyscrasias. METHODS: The serum concentration of soluble syndecan-1 was measured using human syndecan-1 enzyme linked immunosorbent assay kit. RESULTS: Patients with multiple myeloma showed a significantly higher median serum syndecan-1 level than patients with plasmocytoma or monoclonal gammopathy of undetermined significance. Statistically significant differences were also observed among Salmon-Durie subgroups of 50 patients suffering from multiple myeloma. In addition to these findings a statistical correlation with other independent prognostic factors such as serum beta2-microglobulin level, monoclonal immunoglobulin concentration, and bone marrow plasma cell count could also be noted. A significant decrease in median serum syndecan level was observed in patients who responded to chemotherapy, whereas no change in the median syndecan-1 level could be observed in nonresponders. CONCLUSION: These findings confirm the observation that high serum soluble syndecan-1 level is associated with a more advanced disease stage and is a strong independent indicator of poor prognosis. A diminished serum syndecan-1 reading as a result of chemotherapy may be a good indicator of favorable response to antitumor treatment. PMID- 15751513 TI - [Osseous signs of anemia and nutrition disorders in a Medieval cemetery in Hungary]. PMID- 15751514 TI - [Zoltan Krisar M.D. (1914-1990). Memory of a surgeon from Transsylvania]. PMID- 15751515 TI - Coronary slow flow phenomenon: an angiographic curiosity. PMID- 15751516 TI - Robotically assisted cardiac surgery. PMID- 15751517 TI - Robotically enhanced coronary artery bypass surgery. AB - BACKGROUND: Robotically enhanced telemanipulation surgery is a fast developing technique which allows totally endoscopic cardiac surgery with utmost precision and perfection on both beating heart as well as arrested heart. METHODS AND RESULTS: Between December 2002 and February 2004, 125 patients underwent robotically enhanced coronary artery bypass surgery using the da Vinci telemanipulation system (Intuitive Surgical Inc., California). Eleven patients underwent totally endoscopic coronary artery bypass surgery. Of them 9 were done on beating heart while 2 were done on arrested heart. One hundred and fourteen patients had endoscopic takedown of internal mammary artery followed by minimally invasive direct coronary artery bypass in 63 patients and left anterolateral thoracotomy in 51 patients. The internal mammary artery mobilization time was 42 min (35-74 min) while the left internal mammary artery to left anterior descending artery anastomosis time ranged from 20 to 36 min for the totally endoscopic coronary artery bypass patients. In 1 patient, the right internal mammary artery was anastomosed to diagonal artery totally endoscopically. The mean internal mammary artery flow by Doppler measurement done in patients undergoing minimally invasive direct coronary artery bypass was 64 ml/min. Seven patients required conversion to median sternotomy and coronary bypass surgery on beating heart. The mean intensive care unit stay was 1.2 days and the mean hospital stay 4.5 days. There was 1 in-hospital mortality. All 11 patients who underwent totally endoscopic bypass surgery had coronary angiography done at 3 months interval which showed 100% patency in 10 patients while one patient had 50% anastomotic narrowing for which coronary angioplasty was done in the same sitting. CONCLUSIONS: Using telematic technology, a complete endoscopic anastomosis is possible in both single vessel and suitable double vessel disease patients. The use of robotics is now extended to achieve complete myocardial revascularization by harvesting both the internal mammary arteries and making a small thoracotomy for direct anastomosis as well. PMID- 15751518 TI - Predictors of left atrial appendage clot: a transesophageal echocardiographic study of left atrial appendage function in patients with severe mitral stenosis. AB - BACKGROUND: The purpose of this study was to prospectively evaluate a large group of consecutive, non-anticoagulated patients with severe rheumatic mitral stenosis and to analyze the left atrial appendage function in relation to left atrial appendage clot and spontaneous echo contrast formation. METHODS AND RESULTS: We prospectively studied left atrial appendage function in 200 consecutive patients with severe mitral stenosis who underwent transesophageal echocardiography and correlated it with spontaneous echo contrast and left atrial appendage clot. The mean age was 30.2 +/- 9.4 years. Fifty-five (27.5%) patients were in atrial fibrillation. Left atrial appendage clot was present in 50 (25%) patients and 113 (56.5%) had spontaneous echo contrast. The older age, increased duration of symptoms, atrial fibrillation, spontaneous echo contrast, larger left atrium, depressed left atrial appendage function and type II and III left atrial appendage flow patterns correlated significantly (p<0.05) with the left atrial appendage clot. Left atrial appendage ejection fraction was significantly less in patients with clot (21.8 +/- 12.8% v. 39.1 +/- 13.2%, p<0.0001) and in those with spontaneous echo contrast (30.3 +/- 16.2 % v. 40.3 +/- 11.8%, p<0.001). Left atrial appendage filling (18.0 +/- 11.7 v. 27.6 +/- 11.8 cm/s, p <0.0001) and emptying velocities (15.4 +/- 7.0 v. 21.5 +/- 9.6 cm/s, p<0.001) and filling (1.4 +/- 1.0 v. 2.5 +/- 1.4 cm, p<0.0001) and emptying (1.5 +/- 1.2 v. 2.1 +/- 1.2 cm, p <0.05) velocity time integrals were also significantly lower in patients with clot as compared to those without clot. On multivariate regression analysis, atrial fibrillation (odds ratio 6.68, 95% CI 1.85-24.19, p=0.003) and left atrial appendage ejection fraction (odds ratio 1.06, 95% CI 1.00 - 1.11, p=0.04) were the only two independent predictors of clot formation. Incidence of clot was 62.59% in patients with left atrial appendage ejection fraction < or = 25% as compared to 10.4% in those having left atrial appendage ejection fraction >25%. Similarly patients with spontaneous echo contrasthadlower filling (21.7 +/- 11.5 v. 29.4 +/- 12.7 cm/s, p<0.0001) and emptying (17.0 +/- 8.1 v. 23.9 +/- 10.9 cm/s, p<0.0001) velocities, as well as filling (1.9 +/- 1.3 v. 2.7 +/- 1.3 cm, p<0.01) and emptying (1.7 +/- 1.0 v. 2.3 +/- 1.4 cm, p<0.01) velocity time integrals as compared to patients without spontaneous echo contrast. In a subgroup of the patients with normal sinus rhythm, the left atrial appendage ejection fraction was significantly less in patients with clot compared to those without clot (31.2 +/- 13.2 v. 41.3 +/- 11.5 %, p<0.01). CONCLUSIONS: In the patients with severe mitral stenosis, besides atrial fibrillation, a subgroup of patients in normal sinus rhythm with depressed left atrial appendage function (left atrial appendage ejection fraction < or = 25%) had a higher risk of clot formation in left atrial appendage and these patients should be routinely anticoagulated for prevention of clot formation. PMID- 15751519 TI - QT dispersion and early arrhythmic risk in acute myocardial infarction. AB - BACKGROUND: This study sought to find out QT dispersion in healthy individuals and patients of acute myocardial infarction and to find correlation, if any, between QT dispersion and the incidence of ventricular arrhythmias in acute myocardial infarction. METHODS AND RESULTS: QT dispersion was calculated from a 12-lead electrocardiogram in 100 patients of acute myocardial infarction admitted in intensive coronary care unit and 100 age- and sex-matched healthy individuals. In patients of acute myocardial infarction, QT dispersion was calculated on admission, 24 hours after admission and at the time of discharge from intensive coronary care unit. Average QT dispersion in acute myocardial infarction was found to be significantly higher on admission (76.4 +/- 18.3 ms), 24 hours after admission (62.88 +/- 17.52 ms) and at the time of discharge from intensive coronary care unit (51.79 +/- 16.79 ms) than in healthy individuals (29.76 +/- 6.06 ms; p<0.05). QT dispersion was found to be significantly increased in patients of acute myocardial infarction with ventricular arrhythmias (82.06 +/- 16.86 ms) than in those without (66.75 +/- 16.28 ms; p<0.01). Patients of acute myocardial infarction with ventricular tachycardia or ventricular fibrillation had significantly increased QT dispersion (96.25 +/- 15.97 ms) than those who had only ventricular premature beats (80 +/- 15.04 ms; p<0.01). QT dispersion was found to be significantly greater in patients with anterior wall acute myocardial infarction (79.80 +/- 18.19 ms) than in those with inferior wall acute myocardial infarction (71.9 +/- 17.48 ms; p<0.05). At the time of discharge from intensive coronary care unit no statistically significant difference was found in QT dispersion in those who received thrombolysis (51.58 +/- 16.05 ms) and those who did not (48.18 +/- 14.68 ms; p>0.05). QT dispersion was found to be significantly higher in those who died (88.66 +/- 15.97 ms) than in those who survived (74.23 +/- 17.91 ms; p<0.05). QT dispersion was significantly higher in ventricular arrhythmic deaths (97.14 +/- 17.04 ms) than those who had non-arrhythmiac deaths (81.25 +/- 11.25 ms; p<0.05). CONCLUSIONS: Interlead QT variation and its measure as QT dispersion challenges our current approach to the electrocardiographic assessment of arrhythmic risk. QT dispersion may provide a potentially simple, cheap, non-invasive method of measuring underlying dispersion of ventricular excitability. PMID- 15751520 TI - Association of common carotid intima-media thickness and lipoprotein(a) with coronary artery disease. AB - BACKGROUND: Carotid artery intimal medial thickness is a simple, non-invasive and reproducible clinical tool to evaluate atherosclerosis and predict coronary artery disease. Lipoprotein(a) levels are related to both atherogenesis and thrombogenesis and may be a key link between lipid and coronary artery disease. This study evaluated the association of carotid intimal medial thickness and lipoprotein(a) with coronary artery disease. METHODS AND RESULTS: We studied 185 randomly selected patients hospitalized for coronary angiogram in our institute. There were 110 angiographically proven patients of coronary artery disease with mean age of 55.8 +/- 9 years (range 34-72 years) and 75 subjects with normal coronary artery anatomy with mean age of 54.8 +/- 8 years (range 34-68 years). The mean carotid intimal medial thickness of subjects with coronary artery disease was significantly higher than in subjects without coronary artery disease (0.84 +/- 0.16 mm v. 0.65 +/- 0.15 mm, p<0.001). The mean carotid intimal medial thicknesses in patients with triple vessel, double vessel and single vessel disease were 0.96 +/- 0.12 mm, 0.84 +/- 0.11 mm and 0.78 +/- 0.13 mm, respectively (p=0.05). The mean lipoprotein(a) of subjects with coronary artery disease was significantly higher than in subjects without coronary artery disease (35.9 +/- 22.3 mg/dl v. 19.1 +/- 21.2 mg/dl, p<0.001). Mean lipoprotein(a) levels in subjects with carotid intimal medial thickness <0.80 was 26.4 +/- 24.2 mg/dl and in subjects with carotid intimal medial thickness > or = 0.80 was 32.1 +/- 22.1 mg/dl (p=0.05). CONCLUSIONS: There is a strong correlation between carotid and coronary atherosclerosis and carotid intimal medial thickness is a good predictor of presence and extent of coronary artery disease. Lipoprotein(a) level is a powerful independent risk factor for atherosclerosis. Carotid intimal medial thickness and lipoprotein(a) in conjoint can predict coronary artery disease reliably. PMID- 15751521 TI - High prevalence of multiple coronary risk factors in Punjabi Bhatia community: Jaipur Heart Watch-3. AB - BACKGROUND: Studies among emigrant Indian populations have shown a high prevalence of obesity and many coronary risk factors in Bhatia community. To determine the prevalence of risk factors in this community within India we performed an epidemiological study. METHODS AND RESULTS: An ethnic-group sample survey to determine prevalence of cardiovascular risk factors was performed using community registers for enrollment. Methodology used was similar to Jaipur Heart Watch studies performed in 1995 and 2002. We invited 600 randomly selected subjects listed in Punjabi Bhatia community registers and could examine 458 (76.7%) persons (men 226, women 232). Evaluation for coronary risk factors, anthropometric measurements, blood pressure, electrocardiogram, fasting blood glucose and serum lipids was performed using standard definitions. Mean age was 43.2 +/- 14.6 years in men and 44.7 +/- 15.3 years in women. In both men and women there was a high prevalence of family history of coronary heart disease in 45 (19.9%) and 50 (21.6%), family history of diabetes in 96 (42.5%) and 77 (33.2%), sedentary habits in 82 (36.3%) and 73 (31.5%), smoking or tobacco use in 59 (26.1%) and 4 (1.7%), overweight or obesity (body mass index > or = 25 kg/m2) in 123 (54.0%) and 161 (69.4%), severe obesity (body mass index >30 kg/m2) in 47 (20.8%) and 75 (32.3%), truncal obesity (waist-hip ratio: men >0.9, women >0.8) in 175 (77.4%) and 186 (80.2%), increased waist (waist size: men >102 cm, women >88 cm) in 78 (34.5%) and 129 (55.6%), hypertension (blood pressure > or = 140/90 mmHg) in 116 (51.3%) and 120 (51.3%), diabetes in 40 (17.7%) and 33 (14.2%), hypercholesterolemia (total cholesterol > or = 200 mg/dl) in 75 (33.2%) and 67 (28.9%), high triglycerides in 55 (24.3%) and 34 (14.7%), low high-density lipoprotein cholesterol in 169 (74.8%) and 155 (66.8%), and the metabolic syndrome (defined by American National Cholesterol Education Program) in 84 (36.2%) and 111 (47.8%) respectively. Body mass index correlated significantly with (age-adjusted r2 value--men, women) waist diameter (0.52, 0.12), waist-hip ratio (0.21, 0.10), truncal obesity (0.54, 0.60), systolic blood pressure (0.19, 0.16), diastolic blood pressure (0.12, 0.16), hypertension (0.19, 0.31), and metabolic syndrome (0.28, 0.44) (p<0.05). There was a significant linear relationship of body mass index with the prevalence of hypertension, hypercholesterolemia, diabetes (women), and the metabolic syndrome (chi2 for trend p<0.05). Prevalence of these risk factors was the lowest in subjects with body mass index <20 kg/m2. A multivariate ordinal logistic regression analysis revealed that obesity was independently associated with multiple risk factors characterized by metabolic syndrome after adjustment for age, hypertension, and diabetes in both men (odds ratio 2.45, 95% confidence intervals 1.69, 3.57) as well as in women (odds ratio 2.93, 95% confidence intervals 1.86, 4.61) (p<0.01). CONCLUSIONS: There is a high prevalence of obesity, abdominal obesity, hypertension, diabetes, lipid abnormalities and the metabolic syndrome in this community that is significantly greater than reported studies in Jaipur and urban populations elsewhere in India. Obesity correlates strongly with multiple coronary risk factors of which it is an important determinant. PMID- 15751522 TI - Plasma endothelin-1, homocysteine and serum nitric oxide values in patients with left-to-right shunt. AB - BACKGROUND: This study aimed to evaluate the effect of pulmonary blood flow and pulmonary hypertension on plasma endothelin-1, homocysteine and serum nitric oxide levels in patients with left-to-right shunt lesions with pulmonary hypertension and also with normal pulmonary arterial pressure. METHODS AND RESULTS: Plasma endothelin-1, homocysteine and nitric oxide levels were measured in 44 patients (Group 1) with left-to-right shunt and normal pulmonary arterial pressure (Qp/Qs: 2.1), 65 patients (Group 2) with left-to-right shunt and pulmonary hypertension (Qp/Qs: 2.4), 20 healthy control subjects (Group 3), and 17 post-operative patients (Group 4). Plasma endothelin-1 and serum nitric oxide levels were significantly higher in Group 2 than in groups 1, 3, and 4 (p<0.001). Plasma homocysteine levels were significantly higher in Group 2 than in Groups 1 and 4 (p<0.001 and p<0.01, respectively). CONCLUSIONS: The increase in serum nitric oxide levels in patients with left-to-right shunt and pulmonary hypertension may be attributed to the compensatory mechanism. However, this increase does not improve pulmonary hypertension because of increased endothelin 1 and homocysteine levels. In the light of present study, we conclude that vascular changes caused by increased homocysteine and endothelin-1 may provoke pulmonary hypertension in patients with left-to-right shunt. PMID- 15751523 TI - Percutaneous transluminal angioplasty in an embolic middle cerebral artery occlusion. AB - A young lady developed left middle cerebral artery embolism after percutaneous transseptal mitral commissurotomy. She was successfully treated with immediate percutaneous transluminal angioplasty. There was complete recovery of neurological deficit. PMID- 15751524 TI - Transcatheter guidewire perforation of the pulmonary valve as a palliative procedure in pulmonary atresia with intact interventricular septum. AB - We report two cases of pulmonary atresia with intact interventricular septum in whom we successfully perforated and performed balloon dilation of the atretic pulmonary valve using a simple guidewire technique. The technical challenges of performing this procedure in small infants are highlighted. PMID- 15751525 TI - Left ventricular hydatid cyst with myocardial infarction in a patient with severe rheumatic mitral stenosis. AB - Cardiac echinococcosis is rare, and the most serious of all hydatid infestations. We report a case of 30-year-old female who had a hydatid cyst, myocardial infarction and severe rheumatic mitral stenosis. Following mitral valvotomy, the hydatid cyst and the left ventricular aneurysm were totally excised under cardiopulmonary bypass. The patient was discharged on the post-operative day 15 with the advice to continue albendazole for 5 years. PMID- 15751526 TI - Syncope in a middle aged male due to acute rheumatic fever. AB - Rheumatic fever is a multi system disease which occurs following infection with group A beta hemolytic streptococcus. It is commonest in the age group of 5-15 years but can occur in adults also. First degree atrioventricular block is a common manifestation of acute rheumatic fever and is included in the Jones criteria but Wenckebacks phenomena and complete heart block are relatively rare manifestations of rheumatic fever. Syncope occurring in acute rheumatic fever is also infrequently reported. We report the case of a 38-year-old male with rheumatic carditis who had advanced atrioventricular block which resulted in syncope and required a temporary pacemaker insertion. PMID- 15751527 TI - Transatrial permanent pacing lead implantation in a patient of Ebstein's anomaly after one and half repair. AB - Several innovative approaches have been described to achieve endocardial pacing in patients with operated complex congenital heart diseases. We report herein a case of Ebstein's anomaly who underwent a Hardy's repair with a bidirectional Glenn shunt, tricuspid valve annuloplasty and atrial septal defect closure following which she developed complete heart block. The chest was reopened through the previous midsternotomy and a screw-in lead implanted transatrially that resulted in optimal pacing thresholds. This technique offers a viable alternative for endocardial pacing in peri-operative patients requiring permanent pacing. PMID- 15751528 TI - Thrombolysis with percutaneous transluminal balloon angioplasty of a blocked modified Blalock-Taussig shunt. AB - Transcatheter balloon recanalization of occluded Blalock-Taussig shunts in the early post-operative period has been reported in the past but there are issues regarding the role of thrombolysis in this situation. We present our experience with such a procedure in an infant with blocked modified Blalock-Taussig shunt. PMID- 15751529 TI - Is it possible to prevent rheumatic fever? PMID- 15751530 TI - Iatrogenic aortocoronary arteriovenous fistula. PMID- 15751531 TI - Role of cardiac magnetic resonance imaging in identification of amyloid cardiomyopathy. PMID- 15751533 TI - Lipid levels and coronary heart disease: need for region-specific guidelines. PMID- 15751532 TI - Transhepatic balloon dilation of the interatrial septum. PMID- 15751534 TI - Late stent thrombosis following antiplatelet withdrawal while fasting during holy months. PMID- 15751536 TI - Ultrastructural observations of previtellogenic ovarian follicles of dove. AB - Dove ovarian follicle is a complex structure composed of oocyte surrounded by a somatic compartment consisting of theca externa, theca interna and granulosa. The structure of ovarian follicle (1 and 2 mm) of dove was studied by electron microscopy. The granulosa was pseudostratified in the 1-mm-diameter follicles and stratified with two or three irregular rows of cells in the 2-mm-diameter follicles. In the larger follicle indentations between oocyte and granulosa cells become more numerous and the microvilli of granulosa cell elongated to form a zona radiata with similarly elongated oocyte microvilli. Lining bodies were present at the tips of granulosa microvilli and in the cortical region of the oocyte. In the oocyte cortex were observed coated pits, coated vesicles, dense tubules, multivesicular bodies and primordial yolk spheres. Primordial yolk spheres may contain lining bodies and were observed fused with dense tubules and multivesicular bodies or associated with smooth cisternae. PMID- 15751535 TI - Cysteamine, glutathione and ionomycin treatments improve in vitro fertilization of prepubertal goat oocytes. AB - The aim of this study was to improve in vitro embryo development of prepubertal goat oocytes by studying the effect of adding cysteamine to in vitro maturation medium, glutathione (GSH) to in vitro fertilization medium and ionomycin to the sperm capacitation medium. In experiment 1, we analysed the effect of 1 mM GSH added to fertilization medium of oocytes matured with 400 microM cysteamine. The control group were oocytes without cysteamine and GSH. In experiment 2, oocytes matured and fertilized in the presence of 400 microM cysteamine and 1 mM GSH, respectively, were inseminated with spermatozoa treated with ionomycin or heparin. In experiment 1, the percentages of total and normal fertilized oocytes were significantly higher for oocytes supplemented with cysteamine and GSH (40.26% and 30.20%, respectively) than for oocytes from the control group (16.66%, and 10.61%, respectively). The percentage of total embryos obtained after 7 days of culture was significantly higher in the group supplemented with cysteamine and GSH (30.62%) than in the control group (8.09%). In experiment 2, percentages of total and normal fertilized oocytes were significantly higher for the group of spermatozoa capacitated with ionomycin (52.21% and 37.17%, respectively) than with heparin (38.62% and 28.35%, respectively). After 7 days of culture, total embryo rate was significantly higher in the group of sperm capacitated with ionomycin (44.91%) than with heparin (38.69%). However, the percentage of embryos developed to the blastocyst stage was not affected by any of the treatments studied. PMID- 15751537 TI - Effects of melatonin treatment on Leydig cell activity in the testis of the frog Rana esculenta. AB - This study was conducted to verify the effect(s) of melatonin treatment on frog Leydig cells. Morphological observation after melatonin treatment indicates that many frog Leydig cells show degenerative changes (i.e. heterochromatic nuclei, loss of cellular adhesion) while in adjacent germinal tubules several Sertoli cells show heterochromatic nuclei, confirming the presence of a paracrine effect between interstitial and germinal compartments. The effect of melatonin on frog Leydig cell steroidogenesis was investigated in in vitro experiments; after 6 h of incubation melatonin severely inhibits both control and GnRH-induced testosterone secretion. In addition, in order to verify the effect of indolamine on frog Leydig cell activity, we investigated, by in situ hybridization, the presence of frog relaxin (fRLX, a transcript specifically expressed by these cells) in the testes of melatonin-injected animals after 48 h. fRLX signal completely disappeared from the testis of melatonin- injected frogs. The results of the present study indicate that melatonin treatment provokes Leydig cell morphological changes, blocks GnRH-antagonist-induced testosterone secretion and decreases fRLX expression. Taken together these results strongly indicate that melatonin acts on Leydig cells in the testis of the frog Rana esculenta. PMID- 15751538 TI - Ovarian response in sheep superovulated after pretreatment with growth hormone and GnRH antagonists is weakened by failures in oocyte maturation. AB - The administration of growth hormone (GH) or GH plus GnRH antagonists (GnRHa) in sheep allows the enhancement of the pool of gonadotrophin-responsive follicles present in the ovaries and may be useful to increase yields obtained in embryo programmes. The objective of the current study was to evaluate the ability of follicles recruited in response to treatment with GH and GnRHa to grow in response to exogenous follicle stimulating hormone (FSH) and the competence of their oocytes to resume meiosis. Seven females were treated with two doses of GnRHa (days 0 and 3) and three doses of 15 mg of GH (days 3, 4 and 5). Thereafter, this group and a second group (n = 7) were treated with three doses of 1.5 ml of FSH 12 h apart. A third group (control; n = 4) did not receive GH/GnRHa or FSH. The mean number of follicles aspirated on day 7 was higher in ewes treated with GH and GnRHa prior to the stimulation with exogenous FSH than in ewes treated with FSH without pretreatment and in untreated control sheep (20.4 +/- 2.6 vs 17.7 +/- 3.9 and 11.5 +/- 0.8, p < 0.05 and p < 0.01, respectively). The number of recovered cumulus-oocyte complexes after follicular aspiration was higher in the GH/GnRHa + FSH group (8.7 +/- 0.9 vs 6.8 +/- 1.3 in FSH group, n.s., and 4.5 +/- 0.8 in control, p < 0.05), but there were no differences found in the resumption of meiosis (63.1 +/- 9.5% for GH/GnRHa + FSH vs 79.5 +/- 6.3% for FSH and 60.0 +/- 8.8% for control). These results indicate that GH and GnRHa would be useful to increase the number of gonadotrophin responsive follicles in the ovary, but adjustment of later FSH treatment allowing further development of follicles may be necessary prior to its use in superovulatory protocols. PMID- 15751539 TI - Detection of rare Leydig cell hypoplasia in somatic cell cloned male piglets. AB - In this investigation, 22 cloned male piglets were obtained by male fetal fibroblast-cell-derived nuclear transfer. Eighteen of the cloned animals died. The two cell lines did not differ significantly with regard to efficiency of live piglet production. The gross anatomy of the testes of male piglets that died was normal. However, one piglet displayed Leydig cell hypoplasia (LCH). No anatomical defects were detected in the testes of other cloned male piglets. TUNEL analysis of the testis with LCH revealed significant apoptosis in the Leydig cells, while apoptosis was rarely detected in Sertoli cells and spermatogonia. In contrast, testes from the remaining 17 piglets that died appeared normal in size, and their Sertoli and Leydig cell numbers were comparable to those in control piglet testes. Although cloned piglets were derived from fibroblasts obtained from the same fetus, phenotypic instability between cells used for the production of somatic cell cloned piglets suggests that abnormalities in male cloned piglets are caused not by technical problems and/or reprogramming effects, but rather by epigenetically and/or genetically damaged cell-specific effects. PMID- 15751540 TI - The effects of chemical enucleation combined with whole cell intracytoplasmic injection on panda-rabbit interspecies nuclear transfer. AB - Conventional methods of somatic cell nuclear transfer either by electrofusion or direct nucleus injection have very low efficiency in animal cloning, especially interspecies cloning. To increase the efficiency of interspecies somatic cell nuclear transfer, in the present study we introduced a method of whole cell intracytoplasmic injection (WCICI) combined with chemical enucleation into panda rabbit nuclear transfer and assessed the effects of this method on the enucleation rate of rabbit oocytes and the in vitro development and spindle structures of giant panda-rabbit reconstructed embryos. Our results demonstrated that chemical enucleation can be used in rabbit oocytes and the optimal enucleation result can be obtained. When we compared the rates of cleavage and blastocyst formation of subzonal injection (SUZI) and WCICI using chemically enucleated rabbit oocytes as cytoplasm recipients, the rates in the WCICI group were higher than those in the SUZI group, but there was no statistically siginificant difference (p > 0.05) between the two methods. The microtubule structures of rabbit oocytes enucleated by chemicals and giant panda-rabbit embryos reconstructed by WCICI combined with chemical enucleation were normal. Therefore the present study suggests that WCICI combined with chemical enucleation can provide an efficient and less labor-intensive protocol of interspecies somatic cell nuclear transfer for producing giant panda cloned embryos. PMID- 15751541 TI - A novel method for isolating spermatid nuclei from cytoplasm prior to ROSI in the mouse. AB - In the current widely used round spermatid injection (ROSI) protocol for the mouse, the spermatid nucleus is separated from most of the cytoplasm before ROSI by drawing a spermatid in and out of a pipette. This results in the highest rate of normal fertilization. However, this separation method is not always consistent and can be time-consuming. An alternative separation method that cuts away the cytoplasm using the tip of an injection pipette was developed. After removing the cytoplasm, ROSI was performed following both post- and pre-activation protocols and development in vitro and in vivo were examined. The new method consistently removed the bulk of the cytoplasm, as shown by quantifying mitochondria. ROSI without the cytoplasm resulted in significantly higher rates of fertilization than ROSI with the cytoplasm into either post- or pre-activated oocytes. Furthermore, the offspring production rates of ROSI without the cytoplasm were also high (50% and 49% for the post- and pre-activation protocols, respectively). This new method for separating the cytoplasm is an alternative way of producing offspring using ROSI. PMID- 15751542 TI - Is there a germ plasm in mouse oocytes? AB - It was found that in the Graafian oocytes of laboratory mice Mus musculus the population of electron-dense bodies contains two patterns of structures. One of these, designated as cortical granules, originated from the Golgi complex and was surrounded by a membrane. The other was discovered as cristae-containing mitochondrial derivatives lacked an outer membrane. It was found that the mitochondrial derivatives underwent progressive condensation and transformed into electron-dense bodies similar to germinal bodies of metazoan animals. Based on examination of Graafian follicle oocytes from 5 female individuals, about 15% of electron-dense bodies were cortical granules. However, about 85% of electron dense bodies were condensing mitochondrial derivatives transforming into electron dense bodies. PMID- 15751543 TI - Effects of cryodevice type and donors' sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage. AB - Germinal-vesicle-stage oocytes enclosed with compact cumulus cell layers (COCs) were recovered from adult or prepubertal minke whale ovaries, and were vitrified in a solution containing 15% ethylene glycol, 15% DMSO and 0.5 M sucrose using either a Cryotop or an open-pulled straw (OPS) as the cryodevice. The post-warm COCs with normal morphology were cultured for 40 h in a 390 mosmol in vitro maturation medium, and oocytes extruding the first polar body were considered to be matured. The proportion of morphologically normal COCs after vitrification and warming was higher when the COCs were cryopreserved by Cryotop (adult origin, 88.4%; prepubertal origin, 80.8%) compared with the OPS (adult origin, 67.7%; prepubertal origin, 64.2%). The oocyte maturation rate was higher in the adult/Cryotop group (29.1%) compared with those of the prepubertal/Cryotop group (14.4%), the adult/OPS group (14.3%) and the prepubertal/OPS group (10.6%). These results indicate that the Cryotop is a better device than the OPS for vitrification of immature oocytes from adult minke whales. PMID- 15751544 TI - Chromosomal analysis of mouse spermatozoa following physical and chemical treatments that are effective in inactivating HIV. AB - Human immunodeficiency virus (HIV) can be inactivated by heating at 56 degrees C for 30 min, treating with 50% ethanol at room temperature for 10 min, or treating with 2% sodium hypochlorite solution (NaClO) at room temperature for 60 min. Using a mouse model, we evaluated the risk of generating chromosome damage in spermatozoa following these treatments. The spermatozoa were all dead after the treatments. Although 41.3% of oocytes injected with ethanol-treated spermatozoa successfully activated, none of the oocytes injected with heated or NaClO-treated spermatozoa activated. When artificial stimulation with strontium was used, the fertilization of oocytes with heated or ethanol-treated spermatozoa was completely rescued. Sperm nuclei treated with NaClO neither decondensed nor developed to a male pronucleus. The incidences of structural chromosome aberrations in 1-cell zygotes derived from the heated spermatozoa (45.6%) and ethanol-treated spermatozoa (91.2%) were significantly higher than those in the matched controls (5.5% and 10.5%, respectively). Further study is needed to develop a methodology for the protection of spermatozoa against chromosome damage or the separation of damaged spermatozoa before intracytoplasmic sperm injection. PMID- 15751546 TI - The Argentine health system: trends and challenges. PMID- 15751545 TI - Guanylyl cyclase and cGMP-specific phosphodiesterase participate in the acrosome reaction of starfish sperm. AB - In the starfish, Asterias amurensis, the cooperation of three components of the egg jelly, i.e. ARIS (acrosome reaction-inducing substance), Co-ARIS and asterosap, is responsible for inducing the acrosome reaction. Experimentally, ARIS and asterosap are sufficient for the induction. However, when sperm are treated only with asterosap, they become unresponsive to the egg jelly to undergo the reaction. In this study, we analysed the mechanism of the acrosome reaction, using sperm inactivation by asterosap as a clue. Asterosap causes a rapid and transient increase in intracellular cGMP through the activation of the asterosap receptor, a guanylyl cyclase, and causes an increase in intracellular Ca(2+). When sperm were pretreated with asterosap, the guanylyl cyclase seemed to be inactivated irreversibly by dephosphorylation. They were still responsive to ARIS but no longer to asterosap. However, in the presence of IBMX or zaprinast, inhibitors against phosphodiesterases (PDEs), they retained their capacity to undergo the acrosome reaction in response to the egg jelly or ARIS alone. IBMX and zaprinast suppressed the intracellular catabolism of cGMP, but not of cAMP. These results suggest that guanylyl cyclase and cGMP-specific, IBMX- and zaprinast-susceptible PDEs are involved in the regulation of the acrosome reaction. PMID- 15751547 TI - Health and citizenship: the characteristics of 21st century health. AB - Health is at the core of modernity and its governance has been characterised by two expansions: an expansion of the territory of health into an increasing array of personal and political spaces; and an expansion of the do-ability of health. Health is an exemplary area to study the 'consequences of modernity' in all its dimensions and it is inextricably linked to the concept of modern citizenship. PMID- 15751548 TI - Equal future prospects for all hospitals in Europe? Trends in development within the European Union. AB - The Common Market does not stop at the national health systems of the European countries. Hospital systems are no longer compared only among themselves, but they increasingly affect each other and become closely connected. This development concerns the personnel level as well as the exchange of patients. The former president and now vice-president of the German Hospital Association draws a summary of European hospital development over the last few years. PMID- 15751549 TI - The impact on Asian health care systems of nursing migration. AB - Nurses are precious resource in every corner of the world and hospitals are facing serious challenges in providing high quality care with current nursing shortages. While the shortage of nurses is a worldwide issue, impacting currently more on under developed countries, little literature is available on this aspect of the issue. Lots of job opportunities are available for nurses in western countries, advertised on a daily basis, that attract nurses and result in major nursing migration from Asian countries to the western world. PMID- 15751550 TI - Essential emergency surgical, procedures in resource-limited facilities: a WHO workshop in Mongolia. AB - A WHO 'Training of Trainers' workshop on essential emergency surgical procedures was organised in collaboration with the Ministry of Health, Mongolia. The participants included doctors and nurses from the six selected aimags (provinces). Facilitators of the workshop included experts from the Faculty of Health Science University, Mongolian Surgeon's Association and Mongolian Association of Anaesthesiologists association with the Swiss Surgical Team of the International College of Surgeons, Surgical Department of Nurse's College, Trauma Orthopaedic Clinical Hospital, the Department of Quality Assurance of the Directorate of Medical Services, Ministry of Health. Facilitators from the Hospital University of Geneva, Geneva Foundation of Medical Education and Research (RHR) and Evidence and Information for Policy in Geneva, Switzerland participated via video link. The workshop included lectures, discussions, role playing and 'hands on' basic skills training. Videoconference and e-learning sessions using the WHO e-learning tools were conducted at the Global Development Learning Centre. The topics covered during this training workshop included team responsibility and organisation in a health care facility; patient safety; disaster planning; appropriate use of oxygen; management of bleeding, burns and trauma; basic anaesthetic and resuscitation techniques; prevention of HIV transmission; sterilisation of equipment; waste disposal; hygiene; record keeping, monitoring and evaluation of quality of care and checklists prior to surgery to assure that the correct patient gets the correct surgery on the correct side at the correct time. Recommendations were made by the participants on the next steps after this training. PMID- 15751551 TI - The promise of e-health--a Canadian perspective. AB - Canadians value their health care system above any other social programme. Canada's system of health care faces significant financial and population pressures, relating to cost, access, quality, accountability, and the intergration of information and communication technologies (ICTs). The health system also faces certain unique challenges that include care delivery within a highly decentralised system of financing and accountability, and care delivery to a significant portion of the population sparsely distributed across a land mass of 10 million square kilometres, in areas of extreme climatic conditions. All of these challenges are significant catalysts in the development of technologies that aim to significantly mitigate or eliminate these selfsame challenges. The system is undergoing widespread review, nationally and within each province and territory, where the bulk of care provision is financed and managed. The challenges are being addressed by national, regional and provincial initiatives in the public, private and not-for-profit sectors. The promise of e-health lies in the manner and degree to which it can mitigate or resolve these challenges to the health system and build on advancements in ICTs supporting the development of a health infostructure. Canada is actively developing and implementing technological solutions to deliver health information and health care services across the country. These solutions, while exciting and promising, also present new challenges, particularly in regard to acceptable standards, choice of technologies, overcoming traditional jurisdictional boundaries, up-front investment, and privacy and confidentiality. Many organizations and governments are working to address these challenges. Canada Health Infoway, a not-for-profit corporation, was founded by the first ministers in 2001 to accelerate the establishment of an interoperable, pan-Canadian electronic health record. It works with partners in the federal, provincial and territorial jurisdictions to define replicable solutions for establishment of the major elements necessary to achieve this goal. The Canadian Institute for Health Information (CIHI) will also continue to play an increasingly significant role in these initiatives, as the management of health information becomes a more crucial factor in the successful delivery of health care services in the new millennium. PMID- 15751552 TI - The World Health Channel: an innovation for health and development. AB - The issues of the digital divide and of accessing health information in areas of greatest need has been addressed by many. It has been a key component of the discussion of the World Summit for the Information Society and also the focus of an important new initiative, the Global Review for Health Information. Only approximately 1 in 700 people in Africa have internet access compared to a rate worldwide of approximately 10%. Access to essential health information and knowledge management for health care has been deemed a priority for the development of health systems and for the care of patients in areas with limited resources, prompting recent efforts by international organisations and by both governmental and non-governmental agencies (see Godlee et al, 2004 and McConnell, 2004). Health care in developing countries can be limited by many different resources: lack of health care workers with sufficient training, lack of diagnostic equipment, lack of treatment facilities or essential pharmaceuticals; and lack of education or expertise in many relevant areas. Much of the health care done in developing countries is by local lay persons or practitioners or by volunteers working with a variety of NGOs. These volunteers are often very dedicated young people with a vision of health-for-all that is often frustrated in the limited time they are able to spend in these areas and further constrained by meager resources (including availability of appropriate information). The availability of medical expertise and consultation depends largely on the geographical location of the health practitioner and of the patient as well as the level of integration with local practitioners and extent of outside agency involvement. Futhermore, there are often many NGOs working simultaneously on similar projects in the same region without knowledge of each other's activities. Often this occurs simply because a lack of communication exists between organisations, resulting in unnecessary duplication of effort. The availability of medical expertise and consultation depends largely on the geographical location of the health practitioner and of the patient as well as the level of integration with local practitioners and extent of outside agency involvement. The health care worker in developing countries is frequently faced with a paucity of information appropriate to the clinical situations on hand as well as a lack of locally available expertise. The lack of access to health care and other vital resources is one factor in the much lower (by approximately 1/3) life expectancy in the least developed countries campared to industrialised nations. In many developing countries there is only one doctor for 5-10,00 people, compared to a ratio of 1:200 in many developed countries. Textbooks, if they exist, may be 10 20 years out of date and are often directed more at the needs of developed countries. There is thus a growing need for wider availability of training and information on health care in developing countries and support for health care workers. There is also a need for increased communication and collaboration between governmental and non-governmental organisations working in international health to share education, resources and to coordinate efforts in areas supporting improved health care delivery. In recognition of this, the Institute for Sustainable Health Education and Development (www.ished.org) is launching the World Health Channel (WHC) in the spring of 2005 in collaboration with WorldSpace. This will allow access to critical health information in developing countries and place the emphasis on issues important for clinical care for front line health workers in these areas. PMID- 15751553 TI - Global strategy for infection control in hospitals. PMID- 15751554 TI - Major international concerns for 2005. PMID- 15751555 TI - Management of venous thromboembolism: why not treat it at home? PMID- 15751556 TI - Clarification of article on Clostridium difficile--associated colitis. PMID- 15751557 TI - Life cycle of family medicine doctors. PMID- 15751559 TI - Senseless born. PMID- 15751558 TI - Introducing medical students to CAM: response to Oppel et al. PMID- 15751560 TI - Low-molecular-weight heparins during pregnancy. AB - QUESTION: A few years ago I suffered from pulmonary emboli. My physician recommended I use dalteparin during this pregnancy although, during my previous pregnancy, I had received subcutaneous heparin injections three times daily. Is dalteparin the same as heparin? ANSWER: : Based on the best available evidence from mostly small prospective case series, retrospective reports, and placental perfusion studies, low-molecular-weight heparins (LMWHs), such as dalteparin, are a safe and convenient alternative to heparin during pregnancy for both mothers and fetuses. PMID- 15751561 TI - Ophthaproblem. Retinal arteriovenous malformations (type 2). PMID- 15751562 TI - Reviewing a colleague's orders. PMID- 15751563 TI - Practice tips. Managing infected ingrown toenails: longitudinal band method. PMID- 15751564 TI - Rofecoxib (Vioxx) withdrawal: do product monographs adequately inform physicians? PMID- 15751565 TI - Treatment of deep vein thrombosis: what factors determine appropriate treatment? AB - OBJECTIVE: To identify patients with deep vein thrombosis (DVT) for whom in hospital treatment should be considered. QUALITY OF EVIDENCE: The literature was searched for studies on outpatient treatment of DVT. Seventeen studies were assessed: seven were randomized controlled trials (level I evidence), and 10 were non-randomized trials (level II evidence). MAIN MESSAGE: Four criteria can be used to identify patients with DVT for whom outpatient treatment might not be appropriate: presence of massive DVT, presence of symptomatic pulmonary embolism, high risk of bleeding with anticoagulant therapy, and presence of comorbid conditions or other factors that warrant in-hospital care. CONCLUSION: Four criteria can be used to identify patients with DVT for whom in-hospital treatment should be considered. PMID- 15751566 TI - An approach to complex acid-base problems: keeping it simple. AB - OBJECTIVE: To review rules and formulas for solving even the most complex acid base problems. SOURCES OF INFORMATION: MEDLINE was searched from January 1966 to December 2003. The search was limited to English-language review articles involving human subjects. Nine relevant review papers were found and provide th background. As this information is well established and widely accepted, it is not judged for strength of evidence, a is standard practice. MAIN MESSAGE: An understanding of the body's responses to acidemia or alkalemia can be gained through a set of four rules and two formulas that can be used to interpret almost any acid-base problems. Physicians should, however, remember the "golden rule" of acid-base interpretation: always look at a patient's clinical condition. CONCLUSION: Physicians practising in acute care settings commonly encounter acid base disturbances. While some o these are relatively simple and easy to interpret, some are more complex. Even complex cases can be resolved usin the four rules and two formulas. PMID- 15751567 TI - Morbidity and mortality audits: "How to"for family practice. AB - PROBLEM BEING ADDRESSED: While professions hold their members responsible for self-regulation, many physicians have insufficient information about outcome measures in their practices to judge performance and are inexperienced in performing audits to gather the information they need to judge performance. OBJECTIVE OF PROGRAM: To develop a structure and process to support family doctors with little experience in doing quality improvement studies to conduct morbidity and mortality (M&M) audits. PROGRAM DESCRIPTION: A family medicine teaching group provides members on a rotating basis to an M&M review committee. The committee meets eight times a year and has done four audits, the most comprehensive on the topic of preventable hospital admissions. Both implicit and explicit criteria were incorporated into decision making. Strengths and limitations of the audit process and practice changes that resulted from the audit are discussed. CONCLUSION: Morbidity and mortality audits can vary in rigour. To promote physicians' interest in and commitment to audits, factors considered should reflect the goals, needs, skills, and time available of the physicians involved. Practical learning often results from simple projects. PMID- 15751568 TI - Case report: hyperCKemia: a diagnostic dilemma. PMID- 15751569 TI - The meaning of night sweats. PMID- 15751570 TI - Informing policy to control bovine TB: the ISG discusses the wider issues. PMID- 15751571 TI - The foot-and-mouth disease epidemic in Dumfries and Galloway, 2001. 1: Characteristics and control. AB - The foot-and-mouth disease epidemic in Dumfries and Galloway in south-west Scotland comprised 177 infected premises (IPS) in 24 geographical clusters, and ran from March 1 until May 23, 2001. Initial seeding of infection was by livestock (predominantly sheep) that had passed through Longtown Market in adjacent Cumbria. Thereafter, spread within existing, and to new, clusters was associated with the movement of personnel and vehicles, with further transmission by Longtown Market contacts and across common boundaries. Sheep and cattle premises were equally affected. After the peak of the epidemic at the beginning of the third week of March, the upper possible limit of attack rates for premises contiguous to IPS, and premises within 3 km, remained around 10 per cent, with new clusters emerging more distantly. Control procedures included traditional methods of slaughter of all animals on IPS and, elsewhere, of animals considered by veterinary assessment to be Dangerous Contacts; movement restrictions; enhanced biosecurity; tracing of potential sources and spread of virus; and surveillance of premises subsequently considered at risk. These methods were supplemented by the novel pre-emptive slaughter, without veterinary assessment, of all susceptible livestock on all premises contiguous to IPS, and of small ruminants and pigs within a 3 km radius (known as the Protection Zone) around IPS. In total, approximately 80,000 cattle, 564,000 sheep, 2600 pigs and 500 goats were slaughtered, the novel methods accounting for 29 per cent of all cattle and 75 per cent of all sheep killed. Limitations of existing national databases necessitated the development of local databases to administer control procedures. PMID- 15751572 TI - Postdosing colic and diarrhoea in horses with serological evidence of tapeworm infection. PMID- 15751573 TI - Arthrographic diagnosis of shoulder joint masses in two dogs. PMID- 15751574 TI - BVD virus type 2 in British cattle. PMID- 15751575 TI - Congenital myopathy in Beulah speckled face lambs. PMID- 15751576 TI - Draft veterinary medicines regulations. PMID- 15751577 TI - Detection of the Italy O2 strain of infectious bronchitis virus in the UK. PMID- 15751579 TI - Qualitative risk assessment of chronic renal failure development in healthy, female cats as based on the content of eicosapentaenoic acid in adipose tissue and that of arachidonic acid in plasma cholesteryl esters. AB - A study was carried out to assess the qualitative risk of development of chronic renal failure (CRF) in young healthy, female cats as based on the content of arachidonic acid (AA) in plasma cholesteryl esters (CE) and eicosapentaenoic acid (EPA) in adipose tissue. It has been suggested that the content of AA in CE should be <10% of total fatty acids (TFA) and of EPA in adipose tissue be >1.4% of TFA. Subcutaneous adipose tissue and blood samples were obtained from 48 female cats. There was a statistically significant correlation between linoleic acid content of adipose tissue and that of plasma CE. In all cats the EPA content of adipose tissue was lower than 1.4% of TFA and in 30 cats that of AA in plasma CE was higher than 10% of TFA. The EPA content of adipose tissue and the AA content of plasma CE are determined by the contents of these fatty acids in the diet. It is concluded that the fatty acid composition of cat foods should be determined and that, if deemed necessary, the ingredient composition should be altered so that the content of EPA is raised and that of AA is lowererd. PMID- 15751578 TI - First report on the occurrence of 'Helicobacter heilmannii' in the stomach of rabbits. AB - Gastric Helicobacter spp. have been described in a wide range of animal species, including dogs, cats, primates, swine, cattle and rodents. However, in lagomorphs -more specifically rabbits--gastric Helicobacter infections have never been reported. Biopsy specimens were collected from different stomach regions of 23 rabbits, including 10 pet rabbits, 10 industrial animals and 3 research animals. These were subjected to a PCR assay for the detection of Helicobacter DNA. Identification up to the species level was based on 16S rRNA sequence analysis and a recently developed multiplex PCR. Seven rabbits (four pet, one research animal and two industrial animals) tested positive in the Helicobacter genus specific PCR in the stomach, with the corpus being predominantly positive. H. felis and H. salomonis, hitherto presumed to be naturally hosted by cats and dogs, were detected in three animals and one animal, respectively. One of these animals had been completely devoid of any form of contact with cats or dogs. A H. pullorum/H. rappini-like organism (96% 16S rDNA sequence similarity) was found in an industrially held rabbit. The helicobacters of the two remaining rabbits could not be identified up to the species level. To conclude, this is the first report on the occurrence of Helicobacter spp. in the stomach of rabbits. In view of the fact that H. felis and H. salomonis are put forward as having zoonotic potential, further research is necessary to investigate the implications of these findings not only for the rabbit but also for human health. PMID- 15751580 TI - Altered oxidant-antioxidant profile in canine mammary tumours. AB - Mammary tumours are the most common neoplasms in female dogs. Oxidative stress arising due to overproduction of reactive oxygen species, coupled with altered antioxidant capacities has been implicated in the pathogenesis of all types of cancers. However, the extent of lipid peroxidation and the status of antioxidants in canine mammary tumours have not been investigated. The present study was designed to evaluate the oxidant-antioxidant profile in canine mammary tumours. Lipid peroxidation as evidenced by the formation of thiobarbituric acid-reactive substances, lipid hydroperoxides, and conjugated dienes, as well as the status of the antioxidants superoxide dismutase, catalase, reduced glutathione, glutathione peroxidase, glutathione S-transferase and vitamin C, in tumour tissues of 25 bitches was estimated. Lipid peroxidation in tumour tissues was enhanced compared to the corresponding adjacent uninvolved tissues. This was accompanied by significant elevation in both enzymatic and non-enzymatic antioxidants. This study suggests that upregulation of antioxidants induced by lipid peroxidation confers a selective growth advantage to tumour cells over their adjacent normal counterparts. PMID- 15751581 TI - Disposition kinetics of difloxacin in rabbit after intravenous and intramuscular injection of Dicural. AB - This study investigated the pharmacokinetic behaviour of difloxacin following a single intravenous (i.v.) bolus and intramuscular (i.m.) administration of 5 mg/kg body weight (bw) to rabbits (n = 6). Plasma concentrations were determined in triplicate by agar plate diffusion using E. coli (ATCC 25922) as the test organism. Difloxacin was assayed in plasma to determine its concentrations, kinetic behaviour and systemic availability. Plasma concentration-time data generated in the present study were analysed by non-compartmental methods based on statistical moment theory. Difloxacin was rapidly distributed to the tissues with a steady-state volume of distribution (Vdss) of 1.51 L/kg and the total body clearance (Cltot) was 0.59 L/kg/h. The elimination half-lives after i.v. and i.m. administration were 3.25 h and 3.82 h, respectively. After i.m. administration, difloxacin was rapidly absorbed, with mean peak plasma concentration (Cmax of 3.85 microg/ml achieved at 1.61 h (Tmax) post administration. The extent of plasma protein binding of difloxacin in rabbits was 21.45% and the systemic bioavailability was 95.29%. PMID- 15751582 TI - Studies on haemoglobin polymorphism of two breeds of Iranian sheep and its relationship to concentrations of iron, copper, haemoglobin, haematocrit and RBC number. AB - After electrophoresis on cellulose acetate, two haemoglobin phenotypes were detected in Baloochi and Kordi breeds: AA and AB phenotypes. AA was commonest in two breeds. The incidence of type AB haemoglobin in Baloochi and Kordi breeds was 26.5% (9/34) and 9.5% (2/21), respectively. BB phenotype was not seen in Baloochi and Kordi breeds. In sheep with AB phenotype, haemoglobin B was dominant. The mean +/- SD of the two kinds of haemoglobin in sheep with AB phenotype were haemoglobin B percentage 60.5% +/- 9.04%, haemoglobin B absolute 73.84 +/- 5.44 g/L, haemoglobin A percentage 39.5% +/- 9.04%, haemoglobin A absolute 32.88 +/- 2.89 g/L. There were no significant differences for total haemoglobin, haematocrit, red blood cell (RBC) number, iron and copper levels between breed, sex and age groups and also between sheep with AA phenotype and AB phenotype. Pearson's method showed significant correlations for total haemoglobin with packed cell volume (PCV), RBC number, copper concentration and RBC number with PCV, copper level and PCV with copper amount and copper concentration with iron level (p<0.05). In the Kordi breed, significant correlations were seen for total haemoglobin with PCV, RBC number, copper concentration and PCV with RBC number and RBC number with copper level and copper concentration with iron amount (p<0.05). In the Baloochi breed, significant correlations were detected for total haemoglobin with PCV, RBC number and PCV with RBC number (p<0.05). PMID- 15751584 TI - Comparison of the immunofluorescence assay with RT-PCR and nested PCR in the diagnosis of canine distemper. AB - Two pairs of primers were prepared, both localized within the sequences of the nucleoprotein gene (NP) of canine distemper virus (CDV). A number of experiments were done to optimize the conditions of RT-PCR and nested PCR methods. The nucleic acids of the Onderstepoort, Rockborn, Snyder Hill and Lederle strains of CDV could be detected with these primers. However, they did not react with the sequences of the Edmonston strain of the measles virus. The detection limit for RT-PCR was 10 TCID50 and for nested PCR 0.1 TCID50 of CDV. The RT-PCR was able to demonstrate the nucleic acid of CDV in the blood of all seven puppies vaccinated with a modified live virus. Blood samples of 23 dogs clinically suspected of distemper were examined by RT-PCR combined with nested PCR, and the results were compared with the detection of the CDV antigen in the smears from the mucous membranes by the direct immunofluorescence (IF) test. Of the 23 dogs, 12 were positive in nested PCR, six in the IF assay, and only two in single RT-PCR. It is concluded that nested PCR seems to be the most sensitive method for ante-mortem diagnosis of canine distemper, especially in its subacute or chronic forms. PMID- 15751585 TI - Innovations in musculoskeletal oncologic surgery over the year 2000. AB - It is clear that recent advances in related fields including modern imaging, medical chemotherapy, and advances in materials and bioengineering has enhanced the opportunity for limb sparing surgery. This technique should be considered as a surgical option for these patients, when not at the expense of sound oncological procedure. Significant improvements have been noted over the later part of the 20th Century, and in recent years, advances in molecular oncology have resulted in an explosion of knowledge of the biology of cancer which will result in continued progress in this challenging area. PMID- 15751586 TI - Long-term roentgenographic evaluation of proximal femur prosthesis after tumor resection. AB - The need for a better roentgenographic evaluation of implant behavior in long term prosthetic replacement of the proximal femur was considered. The authors present the results of a group of 13 patients belonging to a wide range of ages, with more than ten-years follow-up after tumor resection, and replacement characterized by bipolar cup and non-cemented stem. A new X-ray film acquisition and evaluation method was devised. Results confirmed the good behavior of bipolar reconstructions in patients of different ages, whose sockets were still intact. In younger patients, the prevalent bipolar movement was in the outer bearing, thus limiting polyethylene particulate debris formation. However, debris formation did not significantly affect the bone/stem relationship as only one loose stem was observed in a bone submitted to radiotherapy. PMID- 15751587 TI - Retrograde intramedullary osteosynthesis of femoral fracture: preliminary results of Maggiore Hospital. AB - The Distal Femoral Nail (DFN), Mathys Medical Ltd. Switzerland) was used in our division for the treatment of fractures of the distal metaepiphysis, and in some cases of the diaphysis, of the femur. A total of 16 cases treated over 30 months, between October 2000 and March 2003 (minimum 6, maximum 61 months), were analyzed retrospectively. A total of 11 metaepiphyseal fractures (65%) and 6 diaphyseal fractures (35%) were treated. Consolidation of the fracture was observed in 15 cases out of 16 (equal to 93.8%) with a mean time of 140 days (minimum 120 days, maximum 240 days) after surgery. Mean flexion of the knee equal to 90 degrees and 3 cases of stiffness with lower mobility at 45 degrees were observed: in 2 cases stiffness was of extra-articular origin caused by exuberant bone callus. There were no iatrogenic complications of the vascular type, nor were there infections, or cases of implant loosening. Shortening (1 cm) in a type B2 fracture (based on the AO classification) was observed. The results we obtained suggest that DFN represents a valid alternative to the open treatment of fractures of the distal metaphysis of the femur; in cases of patients with multiple trauma or in elderly patients where invasiveness is reduced, importance is awarded to less blood loss, a shorter amount of time required for surgery, and early mobilization and weight bearing. PMID- 15751588 TI - The surgical treatment of fractures of the radial head: a comparison between osteosynthesis and capitellectomy. AB - Until the mid-eighties, capitellectomy was the treatment of choice for displaced fractures of the radial capitellum. The observation of complications such as instability of the elbow, proximal migration of the radius, valgus and humerus ulna arthrosis, led us to safeguard the capitellum until the biomechanics of the elbow had been restored. A total of 20 patients treated surgically for Mason type II and III displaced fracture of the radial capitellum were studied retrospectively. The patients were divided into two homogeneous groups, of which group 1 was made up of patients treated by capitellectomy, and group 2 by those treated by osteosynthesis. The patients were seen again at mean follow-up of 44.6 months and submitted to clinical and radiographic evaluation. The clinical results were satisfactory in 80% of cases in group 1 and in 100% of cases in group 2. Radiographic signs of arthrosis were present in 90% of patients who had undergone capitellectomy, and in 20% of those who had undergone osteosynthesis. PMID- 15751589 TI - Intrapelvic migration of the prosthetic acetabular component. AB - The authors propose a preoperative evaluation protocol for cases of dislocation of the prosthetic cup complicated by intrapelvic migration, obtained by studying 20 cases of prosthetic loosening with protrusion of the acetabular component in the pelvis, treated by reimplantation or explantation. In all of the patients, accurate preoperative planning was carried out, because of the considerable frequency of dislocation, compression or damage to the vascular and nervous structures deriving from migration inside the pelvis of the acetabular component. The authors suggest that in all cases of acetabular loosening evaluation involve standard X-rays, bone scan with technethium99 and with marked granulocytes, CT scan. When the cup protrudes in the pelvis, prior to surgery, CT scan with contrast medium will be required, and if the risk of vascular involvement exists, angiography should also be carried out. PMID- 15751590 TI - Clinical evaluation of resurfacing of the patella in posterior-stabilizing total knee replacement. AB - The authors report their experience in the implantation of 108 posterior stabilized cemented total knee replacements, where in 43 cases the patella was not resurfaced (group 1) and in 65 in was resurfaced (group 2). The ability to perform daily activities, the subjective function of the patello-femoral joint and the index of patient satisfaction were evaluated by means of a telephone interview; furthermore, a clinical-radiographic evaluation was carried out in 50 cases. The results of the present study indicate the need of resurfacing the patella using a posterior-stabilized prosthetic design, based on the fact that more joint excursion, a low incidence of complications on the patello-femoral joint, and improved ability to perform daily activities were demonstrated. PMID- 15751591 TI - The Ilizarov method in the treatment of malignant neoplasms of the tibia. AB - A total of 3 malignant neoplasms of the tibia are presented: 1 is a mesenchymal chondrosarcoma of the tibial pylon (male aged 14 years), and 2 are cases of squamous skin carcinoma of the leg with tibial infiltration (1 male and 1 female aged 32 and 64 years, respectively). The resections carried out (16.5 cm on the average) were treated by bone transport. Simple transport was used in the first patient, double in the other two. At the end of transport a second surgical stage involving astragalotibial arthrodesis was performed in the first case, and revision of the junction site of bone segments transported with application of autoplastic bone grafts was performed in the other two. The regenerate obtained was slowly corticalized in the first patient, submitted to various cycles of chemotherapy during the course of distraction. In the other two cases, which were not treated by chemotherapy during distraction, corticalization occurred over a shorter amount of time. Follow-up was obtained after 10 years in the first case, and after 2 years in the other 2 patients. There was no long-term recurrence. PMID- 15751583 TI - Botulinum toxins--cause of botulism and systemic diseases? AB - Toxins of Clostridium botulinum (types A-G) are known as 'neurotoxins', causing the clinically well-known picture of flaccid muscular paralysis. The molecular biological background is the blocking of acetylcholine secretion in neuromuscular junctions by enzymatic cleavage of molecules forming the machinery of exocytosis. Two 'non-neurotoxins' (types C2, C3) are produced by some strains of C. botulinum types C and D. These affect the cytoskeleton by ribosylating actin filaments. All these toxins are used as cell biological tools for the study of specific actions and effects in different eukaryotic cells. Pharmaceutical and molecular biological research has shown their influence on several crucial organs (or cell cultures thereof) of humans and animals (brain and spinal cord, cerebellum, hippocampus, hypophysis, pancreas, adrenal glands, salivary glands and others). Under natural conditions, botulinum toxins may pass the intestinal barrier and circulate in the bloodstream for a certain time. Carriers occurring naturally in food, such as wheat germ agglutinin, digitonin or saponin, and bacterial toxins such as streptolysin O, perfringolysins, C2 toxin or botulinolysin may also form pores in cell walls. They facilitate the entry of botulinum toxins into cells that may not have natural binding receptors. It is concluded that in vivo actions of different botulinum toxins after their entry into the organism may contribute to the onset of different diseases of hitherto cryptogenic origin. Some examples are given and future problems are discussed. PMID- 15751592 TI - Patellofemoral arthritis due to localize nodular synovitis (LNS). AB - We present the case of a 38-year-old female patient who had suffered from a progressively increasing anterior pain in her right knee for almost 10 years. She decided to seek medical advice when the pain became intolerable, not permitting her to perform normal everyday activities. The clinical examination showed evidence of patellofemoral arthritis, but the MRI that followed, apart from confirming the original diagnosis as an "Outerbridge" grade III osteoarthritic lesion, revealed an unidentifiable tumor located in the lateral part of the right patellofemoral joint, that appeared to be like a type of "reactive synovitis". The articular degeneration was treated with arthroscopic saving, followed by complete excision of the tumor and anteromedial tibial tubercle transfer under direct vision. The excised tumor was histologically categorized as a "localized type of nodular synovitis" otherwise known as "giant cell" tumor of synovial origin. The patient has fully recoreved from surgery and 3 years post-operatively she has fully returned to everyday normal activities noting only occasional slight discomfort. PMID- 15751593 TI - MRI in diagnosis of osteoid osteoma of the proximal femur: a potentially deviating aspect. Description of a clinical case. AB - Osteoid osteoma constitutes 10-12% of all benign neoplasms of the bone. The tumor more frequently involves the male sex (male to female ratio 2.1:1) and it may be observed in all age groups, with evident predilection for the second decade of life. All of the skeletal segments may be affected, but the most frequent site is the long bones, in the diaphyseal, metaphyseal and more rarely epiphyseal regions. The lesion is characterized by an osteolytic area, the nidus, which is at times partially calcified, surrounded by an osteosclerotic zone that is more or less accentuated. Clinical suspicion and traditional radiography are essential in diagnostic orientation; usually, further imaging methods are also recommended, such as bone scan with Tc99, CT scan and MRI. This last method allows for easy localization of the lesion, although with a sensitivity that is less than that of the CT scan. Nonetheless, the finding, if not supported by clinical suspicion, may be dangerously deviating and it may orient diagnosis towards a more aggressive disease. PMID- 15751594 TI - Heterotopic ossification after hip surgery in cerebral palsy. AB - Heterotopic ossification is a post-surgery complication occasionally observed in patients with infantile cerebral palsy. In the majority of the cases such lesions are found at the hip after surgery on the skeleton and the soft tissues. At our Institute in the last five years, this complication has been observed four times, in 39 patients, who underwent mainly soft tissue releases because of flexion and adduction contracture of the hip. In these cases, as in a further patient with myelomeningocele, the periarticular heterotopic ossification appeared in the hip after tenotomy of the ilio-psoas at the lesser tronchater associated to other surgical procedures. Conversely, no cases of ossification have been found after tenotomy of the adductors or the gracilis or selective tenotomy of the psoas at the pelvic brim. The exact causes of this complication are still unclear, but after an analysis of the literature and patient history it can be hypothesised that it may be related to the surgical procedures carried out. PMID- 15751595 TI - Beyond urban penalty and urban sprawl: back to living conditions as the focus of urban health. AB - Researchers have long studied urban health, both to describe the consequences of urban living and to design interventions to promote the health of people living in cities. Two approaches to understanding the impact of cities on health have been dominant, namely, urban health penalty and urban sprawl. The urban penalty approach posits that cities concentrate poor people and expose them to unhealthy physical and social environments. Urban sprawl focuses on the adverse health and environmental effects of urban growth into outlying areas. We propose a model that integrates these approaches and emphasizes urban living conditions as the primary determinant of health. The aim of the model is to move beyond describing the health-related characteristics of various urban populations towards identifying opportunities for intervention. Such a shift in framework enables meaningful comparisons that can inform public health activities at the appropriate level and evaluate their effectiveness in improving the health of urban populations. The model is illustrated with two examples from current urban public health practice. PMID- 15751596 TI - Optometry in a hospital setting. AB - This study was designed to determine the extent to which optometry has been integrated within the hospital delivery system in New York State and to determine the types of services rendered by optometrists in governmental and non governmental hospitals. A survey was developed and sent to all hospitals in the State to assess which hospitals provided eye care and whether optometrists were on staff. The number of optometrists affiliated with a hospital has increased from 27% in 1978 to 43% in 2003. Virtually all (90%) of city, county, and federal government hospitals used optometrists as compared to 32% of voluntary hospitals. Optometrists were found to provide a full range of medical ocular services including treatment. Given the cost savings of using optometrists, hospitals should consider integrating this profession into their primary eye care services. PMID- 15751597 TI - Cigarette smoking and lesbian and bisexual women in the Bronx. AB - This study investigated the prevalence of cigarette smoking, smoking patterns, and smoking cessation efforts of Black and Hispanic lesbian and bisexual women from a poor, urban community. One-on-one interviews were conducted with a convenience sample of 130 self-identified Black and Hispanic lesbian and bisexual women from the Bronx, NY. Bivariate statistics were used to determine differences between Black and Hispanic respondents in smoking prevalence, frequency, desire to quit, and impact on family unit. Fifty-five percent of Black respondents and sixty-two percent of Hispanic respondents were current smokers. Hispanics were more likely than Blacks to have a partner (p < 0.04), 2 or more children (p < 0.05), and an asthmatic in their household (p < 0.02). Hispanics were less likely than Blacks to have ever attempted to quit (p < 0.04) and to have made a serious attempt to quit in the past year (p < 0.02). Culturally sensitive interventions are needed to help Hispanic lesbian and bisexual women move from the pre contemplative to action stage of quitting. The large proportion of current smokers requires greater access to effective smoking cessation tools. PMID- 15751598 TI - Using community readiness key informant assessments in a randomized group prevention trial: impact of a participatory community-media intervention. AB - This study examines the role of key informant community readiness assessments in a randomized group trial testing the impact of a participatory community-media intervention (which was also complemented by in-school efforts). These assessments were used to help match communities in random assignment, as a source of formative data about the community, as the basis for a coalition-building workshop, and as an evaluation tool, with a follow-up set of surveys approximately 2 years after the baseline survey. Results of the nested, random effects analysis indicated that the intervention influenced community knowledge of efforts and (at marginally significant levels) improved prevention leadership quality and community climate supportive of prevention efforts. There was evidence that the professional affiliation of informants in some cases had an effect on their assessments, which could be controlled in the analysis. The authors conclude that key informant community readiness assessments can usefully serve to supplement aggregated measures of individual attitudes and behavior (reported elsewhere for this study) in evaluating community-based interventions. PMID- 15751599 TI - Barriers and facilitators of colon cancer screening among patients at faith-based neighborhood health centers. AB - We determined the barriers to and facilitators of colorectal cancer (CRC) screening among two faith-based, inner city neighborhood health centers in Southwestern Pennsylvania. Data from a random sample of patients 50 years and older (n = 375) were used to estimate logistic regression equations to compare and contrast the predictors of four different CRC screening protocols: (1) fecal occult blood test (FOBT) < or = 2 years ago, (2) colonoscopy < or = 10 years ago, (3) lower endoscopy (colonoscopy or sigmoidoscopy) < or = 10 years ago, and (4) any of these screening measures. Racial differences (between African Americans or Caucasians) in type of colon cancer screening were not found. Controlling for covariates, logistic regression equations showed that a physician's support of colon cancer screening was positively associated with the receipt of colonoscopy (OR: 19.47, 95% CI: 5.45-69.54), lower endoscopy (OR: 10.96, 95% CI: 3.77-31.88) and any colon cancer screening (OR: 10.12, 95% CI: 3.36-30.46). Patients who see their physicians more frequently were also more likely to be screened for CRC. Unlike other studies, the faith-based environment in which these patients are treated may explain the lack of racial disparity specific to our measures of CRC screening. PMID- 15751600 TI - The management of type 2 diabetes mellitus in children and adolescents. AB - The medical community faces an emerging epidemic of type 2 diabetes mellitus (DM2) in children and adolescents with a disproportionate increase among certain ethnic groups. DM2 represents one arm of the metabolic syndrome and parallels an increasing prevalence of obesity. The metabolic syndrome includes insulin resistance, hyperlipidemia, and hypertension with a consequent risk of early cardiovascular disease. Thus, treatment of DM2 and the metabolic syndrome poses a challenge for pediatric endocrinologists and represents a huge public health issue. This review presents information about treatment of childhood DM2 with emphasis on indications for the use of insulin in management and normalization of blood glucose. PMID- 15751601 TI - 46,XX patients with congenital adrenal hyperplasia: initial assignment as male, reassigned female. AB - Six 46,XX patients with congenital adrenal hyperplasia (CAH) presented with genital ambiguity, five so severe that initial gender assignment was male. Once diagnosis was realized, parents were involved in evaluation and chose sex re assignment as female. To date, these girls and their parents all indicate satisfaction with their decision for a female sex of rearing. The girls have a female gender identity with behavior characteristics known for females with CAH. Thus, while outcome is satisfactory, it is realized that for most, expression of sexual orientation and adult life adjustments have not yet occurred. PMID- 15751602 TI - Update on the prenatal diagnosis and treatment of congenital adrenal hyperplasia due to 11beta-hydroxylase deficiency. AB - 11beta-Hydroxylase deficiency is a common form of congenital adrenal hyperplasia causing virilization of the female fetus and hypertension. DNA analysis of the gene (CYP11B1) encoding 11beta-hydroxylase has been reported previously to be effective in the prenatal diagnosis of one affected female fetus. In that case, prenatal treatment with dexamethasone resulted in normal female genitalia. We now report five new pregnancies that underwent prenatal diagnosis for 11beta hydroxylase deficiency. In the first family, the proband is homozygous for a T318M mutation and all fetuses from four subsequent pregnancies are carriers. In a second family, the mother is homozygous for a A331V mutation and was started on dexamethasone, but identification of a homozygous normal fetus led to the discontinuation of treatment. In another family, the fetus was a male homozygous for R384Q and treatment was discontinued. Lastly, a novel G444D mutation in exon 8 was identified and proven to reduce 11beta-hydroxylase activity. PMID- 15751603 TI - The natural history and genotype-phenotype nonconcordance of HLA identical siblings with the same mutations of the 21-hydroxylase gene. AB - The correlation of genotype to phenotype in congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency has been investigated thoroughly since the mapping of the CYP21 gene to the short arm of chromosome 6. In most instances, it is possible to accurately predict the phenotype based on genoytpe; however, in a small number of patients, individuals with identical mutations demonstrate variable phenotypes. We report two HLA-identical brothers who represent a striking case of genotype-phenotype nonconcordance in CAH. Molecular genetic analysis showed both patients had mutations in intron 2 and exon 10 of CYP21. Both brothers underwent salt-deprivation tests at similar ages over three separate hospital admissions. Patient 1 was diagnosed with simple virilizing CAH and was able to maintain sodium balance during salt deprivation tests. Patient 2, 3 years younger, was diagnosed with salt-wasting CAH and was unable to maintain sodium balance but progressively increased his aldosterone secretion with age. PMID- 15751604 TI - Immunophenotypic characterisation of peripheral blood lymphocytes in autoimmune polyglandular syndrome type 1: clinical study and review of the literature. AB - Autoimmune endocrinopathies are characterised by an increased number of peripheral blood lymphocytes (PBL) expressing activation/ memory markers on their surface. The aim of this study was to determine whether a similar finding could be detected in a group of 11 paediatric and young adult patients suffering from autoimmune polyglandular syndrome type 1 (APS1), also called autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), as very few data have previously been reported in this field. The control group was made up of 11 sex- and age-matched healthy subjects. Fifteen lymphocyte subsets were compared, in terms of percentage and absolute number, and statistical analysis was performed by the Mann-Whitney test. Measurement of T (CD3+), B (CD19+), natural killer (NK, CD3-CD16/56+), CD4+ and CD8+ T lymphocytes showed that patients with APS1 had a higher percentage and absolute count of T lymphocytes: this was entirely due to the statistically larger CD3+CD4+ fraction. Patients with APS1 also had slightly fewer B and NK lymphocytes, but the difference was negligible. Comparison of CD4+ subpopulations bearing activation and naive/memory antigens (marked by CD69, CD25, anti-HLA-DR, CD45RA and CD45RO) showed that patients with APS1 had generally larger percentages and absolute counts of these subsets: however, only the percentage and absolute size of the CD4+CD25+ subset (p = 0.0354 and p = 0.0151, respectively), and the absolute number of the CD4+ anti HLA-DR+ and CD4+ CD45RO+ subsets (p = 0.0193 and p = 0.0209, respectively) were significantly higher. Interestingly, patients with APS1 also had significantly fewer CD8+CD11b+ and CD3-CD8+ cells. In conclusion, PBL distribution in APS1 resembles that of other autoimmune diseases. Further studies are needed to confirm and possibly extend these data. PMID- 15751605 TI - Final height in patients with constitutional delay of growth and development from tall statured families. AB - Constitutional delay of growth and development occurs not only in children of short statured families but also in children of tall statured families. We represent the final height from 12 boys and 21 girls cared for CDGD. The mean target height for boys was 186.6 cm and for girls 173.6 cm and final heights were 191.2 and 176.9 cm, respectively. The final height of 14 patients exceeded the mean target height by more than 4 cm, nine patients exceeded the familial target height range. Only one boy did not reach the familial target height range. We conclude that CDGD occurs in equal quantities in boys and in girls irrespective of the familial height score and will lead to normal final heights within the familial target height range. In general, no therapy is necessary to alter the progression of CDGD. PMID- 15751606 TI - Constitutional delay of growth and puberty: from presentation to final height. AB - This retrospective study evaluated clinical characteristics of patients with constitutional delay of growth and puberty (CDGP) at presentation, during puberty and at final height. The records of 151 children (105 boys, 46 girls) with CDGP were reviewed and the results were evaluated with respect to findings in healthy Turkish schoolchildren. CDGP was twice as frequent in boys as in girls. Height and weight deficit and short sitting height of the children were evident at presentation and continued up to final height. Mean age of onset of puberty was retarded by 2.5 years in girls and by 3 years in boys. The time between onset of puberty and pubertal growth spurt was shorter in both girls and boys than in the controls. Peak growth velocity was compromised in both girls and boys. Forty-one patients (30 boys, 11 girls) reached final height (FH). Mean FH was shorter than both target height and predicted adult height. The Bayley-Pinneau method was found to be a better predictor of FH than either the Tanner-Whitehouse method or target height. FH also showed correlation with the father's height. There was no effect of testosterone treatment on final height. Height deficit at onset of puberty, shorter duration between onset of puberty and pubertal growth spurt, compromised peak growth velocity and short upper segment due to delayed puberty, are findings which may explain the decreased final height of children with CDGP. PMID- 15751607 TI - Longitudinal investigation of the relationship between breast milk leptin levels and growth in breast-fed infants. AB - BACKGROUND: It has been shown that leptin is present in breast milk and human mammary epithelial cells are able to synthesize leptin. It has been suggested that leptin in human milk might be involved in the regulation of postnatal nutrition and growth. AIMS: To investigate whether there is a relationship between leptin levels in human milk and weight gain in the postnatal period and to compare variations of milk-borne maternal leptin concentrations for small for gestational age (SGA), large for gestational age (LGA) and appropriate for gestational age (AGA) infants. INFANTS AND METHODS: Forty-seven healthy lactating women aged from 17-38 years and their infants were included in the study. The infants were separated into three groups according to birth weight as SGA (n = 11), LGA (n = 14) and AGA (n = 22). All infants were fed with breast milk during the study period. Anthropometric measurements were performed on the 15th day of life and at 1, 2, and 3 months of age, and the body mass index (BMI) of the infants' mothers was calculated. Breast milk leptin levels were analyzed by radioimmunoassay. RESULTS: Breast milk leptin levels were found reduced in the SGA group and increased in the LGA group compared to the AGA group at 15 days of life (13.4 +/- 2.2, 28.5 +/- 4.4 and 18.4 +/- 2 ng/ml, respectively; p <0.05). At 1 month of age, leptin levels in breast milk were significantly lower in the LGA group than in the AGA group (15.5 +/- 4.9, 19.4 +/- 1.7 ng/ml, respectively; p<0.05). There was no difference among the three groups at 2 and 3 months of age (p>0.05). There was a positive correlation between birth Weight and breast milk leptin levels on the 15th day (r = 0.47, p = 0.001). A negative correlation was found between weight gain during the first 15 days and 1 month of life and breast milk leptin levels on the 15th day (r = -0.44, p = 0.002; r = -0.40, p = 0.005, respectively). No relationship could be determined between breast milk leptin levels and BMI of the mothers. CONCLUSION: Maternal milk of SGA, LGA and AGA infants had different leptin levels, especially during the first month of life. More rapid growth was shown in the SGA infants during the first postnatal 15 days compared to AGA and LGA infants, and human milk leptin levels were significantly reduced in the SGA group. However, LGA infants gained more weight during the second 15 days of life and breast milk leptin levels were dramatically decreased in LGA and increased in SGA infants at the end of first month of life. These findings suggest that the presence of leptin in breast milk might have a significant role in growth, appetite and regulation of nutrition in infancy, especially during the early lactation period, and the production of leptin in breast tissue by human mammary epithelial cells might be regulated physiologically according to necessity and state of the infant. PMID- 15751608 TI - Leptin profile in neonatal gonadotropin surge and relationship between leptin and body mass index in early infancy. AB - AIM: This study was designed to investigate the longitudinal and dynamic profile of leptin and its relationship with sex hormones including luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2) and testosterone (TTE) in neonatal 'minipuberty'. We also investigated the effects of leptin in the regulation of body weight gain and body mass index (BMI) in the first 3 months of life. METHODS: A longitudinal study was carried out in a cohort of 15 male and 15 female term infants during the first 3 months of life. Blood samples were collected in the morning from the infants on the 3rd, 15th, 30th, and 90th days of life. At each sample collection, anthropometric measurements were recorded. Serum leptin, LH, FSH, E2 (girls only) and TTE (boys only) concentrations were analyzed using standard biochemical methods. Association of leptin with weight gain, BMI, and these hormones during infancy was evaluated. RESULTS: Leptin levels increased significantly between the 3rd and 90th days of life in both boys and girls. BMI of both groups increased significantly from the 3rd to the 90th day. There was no significant difference in the leptin levels and leptin/BMI ratios of the two sexes at different time points. Leptin levels on the 30th and 90th days correlated significantly with BMI in both sexes. LH and FSH levels in both groups were found to be significantly higher on the 15th day of life. No correlation was observed between leptin and LH, FSH, E2 or TTE levels throughout the study. CONCLUSIONS: Leptin levels do not differ between the two sexes during early infancy and possibly there is no role for leptin in the surge of gonadotropins or sex steroids in neonatal minipuberty. The relationship between leptin and BMI could not be seen in the first postnatal days and the transient lack of the regulatory effect of BMI on leptin concentrations might reflect an adaptive resistance in the production of leptin to support catch-up growth after initial physiological weight loss in newborns. PMID- 15751609 TI - Three new 46,XX male patients: a clinical, cytogenetic and molecular analysis. AB - BACKGROUND: XX males range phenotypically from completely masculinised individuals to true hermaphrodites and include a subset of SRY negative patients. The correlation between genotype (SRY+/-) and phenotype is still unclear. AIM: To report three new patients with this rare condition, one of whom was diagnosed prenatally and another was SRY negative, and to verify in our patients whether the presence of SRY results in a more masculinised phenotype. PATIENTS AND METHODS: We present two phenotypically normal XX male patients (10 and 13.5 years) and one 3.1 years old XX male with ambiguous external male genitalia Prader IV. The patients were diagnosed by clinical, hormonal, sonographic, genetic and histological criteria. RESULTS: Basal hormonal status was normal for phenotype but an excessive response to GnRH testing was noticed in the second patient together with insufficient hCG stimulation in all three patients. Pelvic ultrasound displayed male structures without Mullerian ducts; testicular biopsy, performed only in the intersex patient, showed Sertoli and Leydig cell hypoplasia. Chromosome analysis confirmed 46,XX karyotype. FISH analysis and molecular analysis by PCR were positive for Yp fragments/SRY gene on Xp in two patients and negative in the patient with ambiguous external genitalia. CONCLUSIONS: In our observation Y chromosome-specific material containing the SRY gene translocated to the X chromosome results in a completely masculinised phenotype. In the intersex patient, incomplete masculinisation without SRY suggests a mutation of one or more downstream non-Y testis-determining genes. PMID- 15751610 TI - Growth hormone deficiency in a child with Williams-Beuren syndrome. The response to growth hormone therapy. AB - Pre- and postnatal growth retardation of unknown pathogenesis is a common clinical feature in patients with Williams-Beuren syndrome (WBS). However, growth hormone deficiency (GHD) has not been considered a major cause of growth retardation. There is only one patient in the literature with confirmed GHD who responded well to human growth hormone (hGH) therapy. We report a female infant with confirmed WBS who, through provocative testing, was found to have GHD and who responded satisfactorily to hGH therapy. Height SDS was -4.2 at the age of 12 months when hGH was initiated and increased to -0.8 at the age of 4.25 years. The pathogenesis of GHD in our patient is unclear. Nevertheless, the elevated levels of prolactin and the response of hGH to growth hormone releasing hormone (GHRH) administration are indicative of a hypothalamic rather than pituitary defect. In conclusion, GH deficiency might contribute to the growth failure in a number of patients with WBS and in such cases hGH therapy will most likely improve final height. PMID- 15751611 TI - Novel growth hormone receptor mutation in a Chinese patient with Laron syndrome. AB - Laron syndrome, growth hormone (GH) insensitivity syndrome, caused by a mutation of the GH receptor (GHR) gene, is extremely rare in the Chinese population. We report a Chinese girl diagnosed with Laron syndrome at age 1.9 years with height 4.9 SDS, basal GH 344 mIU/ml, IGF-I <12 ng/ml, IGFBP-3 <0.2 mg/ml, and undetectable GHBP. A novel mutation of the GHR, not previously described, was identified at the donor splice site of intron 6. PMID- 15751612 TI - Primary hyperaldosteronism with normokalaemia secondary to an adrenal adenoma (Conn's syndrome) in a 12 year-old boy. AB - A 12-1/2 year-old boy presented to the Accident Department following an episode of dizziness and was found to be hypertensive. Investigations revealed primary hyperaldosteronism secondary to an adrenal adenoma (Conn's syndrome). He had normal electrolytes during the period of investigation and potassium concentrations were > or = 4.2 mmol/l on all but one occasion. The hypertension resolved following excision of the adrenal tumour. Normokalaemia with potassium >4.0 mmol/l is very unusual in patients with Conn's syndrome and has not been described in childhood before. Primary hyperaldosteronism needs to be considered in hypertensive children even when potassium concentrations are well within the laboratory reference range. PMID- 15751613 TI - Reversed causality--a need to revisit systems modeling of work-stress-health relationships. PMID- 15751614 TI - Different mechanisms to explain the reversed effects of mental health on work characteristics. AB - OBJECTIVES: The number of longitudinal studies reporting evidence for reversed effects of strain on work is growing, but evidence regarding the mechanisms underlying such effects is scarce. In this study, earlier longitudinal findings were reviewed, and the following four mechanisms for reversed effects were proposed that reflect within-person or environmental changes: (i) the rosy perception mechanism, (ii) the gloomy perception mechanism, (iii) the upward selection mechanism, and (iv) the drift mechanism. METHODS: These mechanisms were tested using structural equation modeling and longitudinal data from a Dutch four phase study (N=1588 participants). RESULTS: The results revealed that work characteristics and mental health influenced each other reciprocally and longitudinally. The reversed effects were examined in more detail, and it was found that these could be accounted for by both within-person and environmental change mechanisms. The rosy perception mechanism was found to explain the positive effects from health on job demands; the upward selection mechanism explained the positive (environmental) effects from health on job control; the gloomy perception mechanism explained the reversed (evaluation) effects from health on supervisory social support. No support was found for the drift mechanism. CONCLUSIONS: Mechanisms that may explain reversed causation are yet poorly understood. The main contribution of the present study lies in the fact that it proposes (i) a conceptual framework with which to analyze the effects of health on work characteristics and (ii) methods for testing these mechanisms. The study revealed that there is good reason to pursue research on reversed causality. PMID- 15751615 TI - Disentangling the causal relationships between work-home interference and employee health. AB - OBJECTIVES: The present study was designed to investigate the causal relationships between (time- and strain-based) work-home interference and employee health. The effort-recovery theory provided the theoretical basis for this study. METHODS: Two-phase longitudinal data (with a 1-year time lag) were gathered from 730 Dutch police officers to test the following hypotheses with structural equation modeling: (i) work-home interference predicts health deterioration, (ii) health complaints precede increased levels of such interference, and (iii) both processes operate. The relationship between stable and changed levels of work-home interference across time and their relationships with the course of health were tested with a group-by-time analysis of variance. Four subgroups were created that differed in starting point and the development of work-home interference across time. RESULTS: The normal causal model, in which strain-based (but not time-based) work-home interference was longitudinally related to increased health complaints 1 year later, fit the data well and significantly better than the reversed causal model. Although the reciprocal model also provided a good fit, it was less parsimonious than the normal causal model. In addition, both an increment in (strain-based) work-home interference across time and a long-lasting experience of high (strain-based) work-home interference were associated with a deterioration in health. CONCLUSIONS: It was concluded that (strain-based) work-home interference acts as a precursor of health impairment and that different patterns of (strain-based) work-home interference across time are related to different health courses. Particularly long-term experience of (strain-based) work-home interference seems responsible for an accumulation of health complaints. PMID- 15751616 TI - Total mortality and cause-specific mortality of Swedish shift- and dayworkers in the pulp and paper industry in 1952-2001. AB - OBJECTIVES: The study investigated the relationship between shiftwork and mortality, both total mortality and cause-specific mortality from coronary heart disease (CHD), stroke, and diabetes. Methods The cohort consisted of 2354 shiftworkers and 3088 dayworkers in two pulp and paper manufacturing plants. The mortality of the cohort was monitored from 1 January 1952 to 31 December 2001 by linkage to the national Cause of Death Register. Groups of workers defined by different durations of shiftwork exposure were compared with dayworkers by calculating standardized relative rates (SRR). RESULTS: Death due to any cause (total mortality) was not higher among the shiftworkers than among the dayworkers [SRR 1.02, 95% confidence interval (95% CI) 0.93-1.11]. A longer duration of shiftwork was associated with an increased risk of CHD, and shiftworkers with >30 years of shiftwork had the highest risk of CHD (SRR 1.24, 95% CI 1.04-1.49) Diabetes was more common as the number of shift years of exposure increased [b(linear coefficient) = 4.14 x 10(-5), 95% CI 2.46 x 10(-5) -5.81 x 10(-5)]. Compared with dayworkers, shiftworkers had a greater risk of death due to stroke (SRR 1.56, 95% CI 0.98-2.51). CONCLUSIONS: In the present study, no general increase in mortality was observed among shiftworkers when they were compared with dayworkers. However, the results demonstrate an increased mortality from CHD among shiftworkers with a long duration of shiftwork exposure. Mortality due to diabetes also increased as the number of shift years and mortality due to ischemic stroke in shiftworkers increased. PMID- 15751617 TI - Cancer in the Norwegian printing industry. AB - OBJECTIVES: The aim of this study was to investigate cancer risk among Norwegian workers in the printing industry, particularly lung and bladder cancer. METHODS: Cancer incidence was investigated from 1953 through 1998 in a cohort of 10 549 male members of a trade union in the printing industry in Oslo and nearby areas. Rates from the region, were used to calculate standardized incidence ratios (SIR) separately for the skilled and unskilled workers. Smoking data from a sample of the cohort were utilized for evaluating the risk estimates of smoking-related cancers. Specific exposure data were not available. RESULTS: Among the skilled workers, significantly elevated risks of cancer of the urinary bladder [standardized incidence ratio (SIR) 1.47, 95% confidence interval (95% CI) 1.19 1.79], liver (SIR 1.92, 95% CI 1.15-2.99), pancreas (SIR 1.46, 95% CI 1.07-1.94) and colon (SIR 1.27, 95% CI 1.05-1.55) were observed, whereas an increased risk of lung cancer in this group was confined to those born before 1910. Among the unskilled workers, there were significantly increased risks of cancer of the mouth, esophagus, stomach, larynx, lung, and all sites. CONCLUSIONS: The study showed that workers in the printing industry were at increased risk of several types of cancer. In particular the increased risk of bladder cancer among the skilled workers is suggestive of an occupational cause. However, no specific agent could be identified as an occupational carcinogen. The results did not support the hypothesis of a generally increased risk of lung cancer. The risk pattern for unskilled workers may reflect confounding by nonoccupational factors. PMID- 15751618 TI - Impairment of lung function in asbestos-exposed workers in relation to high resolution computed tomography. AB - OBJECTIVES: The aim of the study was to determine the causes of impairment of ventilatory function and diffusing capacity in smoking asbestos-exposed workers (N=590) showing radiological pleural thickenings or pulmonary fibrosis. METHODS: High-resolution computed tomography (HRCT) and spirometry were performed, and diffusing capacity was measured. The workers were divided into five groups based on the HRCT scoring: pleural disease (N=190), pulmonary fibrosis (N=68), emphysema (N=148), combined fibrosis and emphysema (N=74), and marked adhesions (N=110). The graded lung function impairment was compared between the groups. RESULTS: Moderate impairment of forced expiratory volume in 1 second [odds ratio (OR) 2.72, 95% confidence interval (95% CI) 1.31-5.57] and forced vital capacity (OR 2.81, 95% CI 1.05-6.89) was associated with the persons with combined fibrosis and emphysema. Marked impairment of diffusing capacity was associated with the combined fibrosis and emphysema (OR 4.94, 95% CI 2.48-9.77) but not with pleural disease (OR 0.21, 95% CI 0.09-0.45) or pulmonary fibrosis (OR 0.36, 95% CI 0.08-1.05). For the persons with combined fibrosis and emphysema, the mean fibrosis score did not differ between normal, slightly reduced, or markedly reduced diffusing capacity, but the emphysema score was significantly higher for the patients with marked impairment than for those with normal diffusing capacity (P < 0.01). CONCLUSIONS: Different profiles of asbestos- and smoking-induced pulmonary or pleural disease were found. The results indicate that the most important factor determining the degree of functional impairment in smoking asbestos-exposed workers is the presence of pulmonary emphysema. PMID- 15751619 TI - Nasal mucosal histamine reactivity among teachers six years after working in a moisture-damaged school. AB - OBJECTIVES: The objective of this study was to determine whether verified increased nasal mucosal reactivity among teachers, who had been working in a school with severe moisture problems, still persisted 6 years after remedial measures had been taken. The increased nasal mucosal reactivity, measured as the mucosal swelling reaction upon histamine provocation, had earlier been shown both 1 and 3 years after the renovation. METHODS: Twenty-four teachers in the target school and sixteen teachers in the control school, who participated in all the investigations (1995, 1997 and 2000), answered a standardized questionnaire and underwent a nasal histamine provocation test. In addition to the registration of mucosal swelling, the microcircular reaction to histamine provocation was measured with laser-Doppler flowmetry. RESULTS: The nasal histamine reactivity among the teachers, measured as the mucosal swelling reaction, was no longer increased. However, the laser-Doppler flowmetry showed that the teachers in the target school had significantly divergent microcircular perfusion (P = 0.0022) and a concentration of moving blood cells (P=0.0009) in the histamine provocation when compared with the teachers in the control school; this finding indicates more-pronounced plasma leakage and edema from the nasal mucosa. CONCLUSIONS: Restored nasal histamine reactivity, measured as mucosal swelling reaction, was observed among the teachers 6 years after an obviously successful renovation of the school environment. However, changes in microcirculation indicate a remaining effect on the nasal mucosa. PMID- 15751620 TI - Methacholine bronchial responsiveness and variations in lung function among workers exposed to flour. AB - OBJECTIVES: Methacholine bronchial responsiveness and variations in the pulmonary function of workers exposed to wheat flour and a reference group were compared. METHODS: Each subject [140 men exposed to flour (bakers and pastry makers) and 77 controls] completed a standardized questionnaire. Bronchial responsiveness was quantified by measuring the slope between percentage decrements in forced expiratory volume in 1 second (FEV1) and cumulative methacholine dose. FEV1 and peak expiratory flow (PEF) were recorded four times a day for 15 days using a handheld electronic spirometer. The variability in the FEV1 and PEF readings was expressed as variation coefficients (100 x standard deviation/mean). RESULTS: The mean duration of exposure to flour was 14 (SD 9) years. Rhinitis was significantly more common in the exposed group than in the control group (30.7% versus 11.7%, P = 0.001). The mean FEV1 and PEF did not significantly differ between the two groups. The slope of the dose-response to methacholine and the variation coefficients were lower among the unexposed nonsmokers than among the exposed workers and smokers. The differences were significant for the exposed smokers. The two variation coefficients correlated with each other (r = 0.82) but not with the slope of the methacholine challenge. Conclusions Occupational exposure to flour and smoking increase bronchial responsiveness, as measured by the slope of the dose-response to methacholine and the variation coefficients of airflow. However, methacholine bronchial responsiveness and the variability of lung function do not measure exactly the same aspect of airway behavior. PMID- 15751621 TI - Assessment of exposure to wheat flour and the shape of its relationship with specific sensitization. AB - OBJECTIVES: Dust and wheat-allergen exposure were assessed among bakers, flour millers, and bakery-ingredient producers, and the risk for sensitization was studied. METHODS: About 520 inhalable dust and wheat-allergen measurements were made among 270 Dutch workers. Data on sensitization to wheat and common allergens (atopy) were also available. Exposure was estimated according to the sector of industry, job title, and tasks. The shape of the relationship between sensitization and exposure was studied using a two-stage modeling approach: semi parametric generalized additive modeling and, consequently, a simple description of the relationship using a parametric logistic model. To reduce the effect of exposure measurement errors (attenuation), a combination of the actual measured exposure and variance-weighted estimates of exposure was used. Results The effect of exposure to both inhalable dust and wheat allergens on sensitization was described best by a linear relationship in three industries and a quadratic relationship in one industry. The relation for the whole study population was best described as quadratic, and the probability of sensitization increased with exposure up to -2.7 mg/m3 for inhalable dust and approximately 25.7 microg EQ/m3 for wheat allergens. The risk decreased at higher exposures (P = 0.0121 and P = 0.0731 for dust and wheat, respectively). Atopy and sector of industry modified the sensitization risk significantly in all the analyses. Using a variant weighted estimator to calculate exposures corrected for the bias and resulted in almost the same point risk estimators. Conclusions Exposure-response relationships for allergens may be nonlinear and differ between industries. A threshold is not indicated on which to base occupational exposure standards; alternatively, other approaches, such as benchmarking, seem warranted. PMID- 15751622 TI - Impact of smoking policy on the respiratory health of food and beverage servers. AB - OBJECTIVES: The purpose of this study was to determine whether workplace smoking policy was associated with respiratory health effects among food and beverage servers. METHODS: Data were obtained from a postal survey of hospitality workers. The participation rate for the questionnaire was 73.9% of those contacted. Current smokers were excluded from the analysis. Adjustment for differences between groups in age, gender, ex-smoker versus never smoker status, home exposure environmental tobacco smoke, childhood asthma, mail versus telephone questionnaire, and hours worked per week was done using logistic regression. A subset of 88 nonsmokers underwent laboratory evaluation, including spirometry and hair nicotine analysis. RESULTS: The prevalence of irritant and respiratory symptoms among 383 nonsmokers was consistently higher among the participants from premises where smoking was permitted without restrictions on the workplace. In comparison with those from facilities where smoking was prohibited, the highest adjusted odds ratios (OR) were for chronic phlegm for those working where smoking was permitted (OR 8.5 95% confidence interval (95% CI) 2.4-30.0] or where there were partial smoking restrictions (OR 5.7 95% CI 1.7-19.4). Lung function was not reduced apart from the ratio between forced expiratory volume in 1 second and forced vital capacity, which was lower for workers from facilities where smoking was permitted. Hair nicotine levels were lowest for workers from facilities where smoking was prohibited. CONCLUSIONS: The results suggest that occupational exposure to environmental tobacco smoke, determined through smoking policies, can adversely affect the respiratory health of nonsmokers who work in the food and beverage service industry. PMID- 15751623 TI - [Recent trend of diagnosis and treatment for Zollinger-Ellison syndrome]. AB - Since 1955 it has been difficult to cure gastrinomas in patients with Zollinger Ellison Syndrome (ZES), because gastrinomas exhibit symptoms when they are still small. Besides they are often multiple and metastatic. Development of the SASI test and SRS has changed the situation, and now, based on these two techniques, curative resection of gastrinomas in ZES can be routinely performed. As the cases of resection increase, more accurate pathological findings have accumulated. We know now that duodenal gastrinomas have been a cause of ZES in more patients than pancreatic gastrinomas. In most patients with MEN-1 and ZES, duodenal gastrinomas, which are often multiple, bring about ZES. PMID- 15751624 TI - [Recent changes in informed consent for cancer treatment]. AB - "Truth telling" which was focused on just telling patients they have cancer, is gradually developing into "informed consent (IC)" which is focused on patients' understanding of their disease after careful explanation by their doctors. The major goal of IC is to respect the patients' self-determination and self selection of the therapies they will receive and to establish close relationships between patients and doctors. Although truth telling about patients' prognosis is not yet fully established, providing such information to patients is basically unavoidable to obtain true IC. In delivering the bad news and obtaining IC as part of patient medical care, it is very important for doctors to brush up their communication skills with patients. PMID- 15751625 TI - [Possible approaches to cancer chemotherapy in palliative care medicine]. AB - We discussed our attitude, ideas, and treatment practices, and presented several problems concerning palliative chemotherapy under the palliative medical environment in patients with advanced cancer in our hospital. We thought that, in many cases, they could be given further chemotherapy. Because of the clear availability of so-called palliative therapy, these patients tend not to elect another chemotherapy using more effective anti-tumor agents in the name of "acceptance." The clinical practice of palliative chemotherapy, however, really needs the passion of skillful medical oncologists, palliative care doctors, and other staff including surgeons and radiologists. The most important issue is that all the medical staff sincerely empathize with the wishes of patients and their families with a common hospice spirit. PMID- 15751626 TI - [Parenteral opioids in end-of-life care in cancer patients]. AB - The ethical obligation to relieve pain and other distressing symptoms in patients with cancer is now receiving increasing attention and concern in palliative and end-of-life care. The vast majority of patients with cancer pain are prescribed opioids in oral formulation. However, a patient at the end of his or her life often needs parenteral administration of opioids as the medical condition deteriorates, irrespective of using sedatives. Controlled release opioid preparations currently available in Japan are CR morphine sulphate, CR oxycodone and transdermal fentanyl patch. The opioids available in injectable preparations are morphine HCL and fentanyl citrate. Accordingly, not only the guideline for switching opioids, so-called "opioid rotation," but the guideline for changing the opioid administration route is also necessary in end-of-life care. The authors specifically indicate when and how to convert opioid administration from the oral to the parental route. PMID- 15751628 TI - [Palliative care team at general hospital]. AB - Some three hundred thousand patients die from cancer in a year, most of whom end their lives at a general hospital. Considering this situation, the general hospital has to provide palliative care and make up interdisciplinary Palliative Care Team (PCT). Since April 2002, we established a PCT at Akita City Hospital. Our PCT educated doctors and nurses for palliative care medicine and established a system for palliative care. For example, the assessment sheet of cancer pain is useful for treatment of pain and to equally recognize the patient's condition by each team. The PCT provides support for doctors and nurses in the general unit. We general doctors must learn more about how to provide palliative care medicine for at least cancer pain as well as cancer treatment. A team approach is needed for all cancer patients. PMID- 15751627 TI - [Pain management for cancer patients with critical pathway on computer]. AB - For relief from cancer pain, we developed critical pathway (CP) as an effective strategy for the medical staff treating cancer patients. This CP was made out of Microsoft Excel, and was used on personal computers. "Good sleeping" was set as the first goal and the second was "No pain in rest position." To achieve this, physicians and nurses evaluate medical efficacy and complications including nausea/vomiting, constipation, somnolence and hallucination everyday using controlled release oxycodone in addition to NSAIDs and prochlorperazine, stool softener and peristaltic stimulant for adverse effects. These outcomes lead to the medication change the next day by calculation using visual basic function due to opioid titration theory. In twelve patients this CP was acceptable, and all of them achieved the second goal within a week without severe adverse effects except constipation. PMID- 15751629 TI - [Palliative care in Japan--current status and future]. AB - This review paper identifies the current challenges in palliative care in Japan and proposes multiple areas to be improved in future, including respect for the individuality of patients and families, early referrals to specialized palliative care service, availability of medical treatments in specialized palliative care facilities, quality control of specialized palliative care systems, establishment of regional palliative care programs, dissemination of education programs at the various levels, and organizing a multicenter research network. PMID- 15751630 TI - [Assessment of dihydropyrimidine dehydrogenase and thymidylate synthase expression in gastric carcinoma and colonic carcinoma]. AB - We measured DPD activity and TS content in cancer and normal mucosa obtained from 23 patients with colorectal carcinoma and 7 patients with gastric carcinoma. DPD activity in colorectal carcinoma cases was significantly decreased in cancer tissues compared to those in adjacent normal mucosa (p<0.001), while TS content was significantly increased in cancer tissues (p<0.01). No significant correlation was found between cancer and adjacent normal mucosa in DPD activity and TS content of gastric carcinoma cases. Immunohistochemically, DPD and TS overexpression were found in 11 out of 23 patients (47.8%) and 3 out of 23 patients (13%) in colorectal carcinoma cases, respectively. In gastric carcinoma cases, DPD and TS overexpression were found in 3 out of 7 patients (42.9%) and 3 out of 7 patients (42.9%), respectively. No significant correlation was found between quantitative measurement and immunohistochemical overexpression in colorectal and gastric carcinoma cases. DPD and TS protein were partially overexpressed in some cases. Heterogeneity of the tumor is thought to be one of the reasons for the discrepancy between quantitative measurement and immunohistochemical overexpression. PMID- 15751631 TI - [Trial of outpatient anti-cancer chemotherapy with infusion of 5-FU and cisplatin for advanced gastric and colorectal cancers]. AB - We carried out a pilot study on the clinical efficacy and safety of outpatient anti-cancer chemotherapy with 5-FU and CDDP for 5 patients with advanced cancer of the stomach or colon, using two disposable balloon pumps. The protocol was combined chemotherapy with continuous intravenous infusion of 5-FU (500 mg/body/day) and CDDP (10 mg/body/day) in 5-day courses for 1 week, and the therapy was repeated as long as possible. Pharmacokinetic study showed that the mean serum concentration of 5-FU was 64.3+/-9.2 ng/ml, and the serum concentration of total Pt increased continuously during CDDP injection. Thus, both drugs were injected, safely and surely. One patient had a clinically evaluable lesion, and the anti-tumor effect of this case was SD. But the serum CEA level was decreased in 3 cases. The side effect of Grade 3 and 4 was not seen, but nausea, vomiting, anorexia, and weight loss were observed frequently. This therapy enabled the patients to stay home 51.6+/-10.0 days longer than with the usual methods in hospital, and this therapy was thought to improve their quality of life. Thus, this therapy is feasible and quite useful, but much attention must be paid to the patient's oral uptake during the therapy, and the clinical effects should be evaluated in randomized control trials. PMID- 15751632 TI - [Results of treatment of far advanced and recurrent stomach cancer with TS-1]. AB - We used TS-1 as first-line therapy to treat 44 patients with far advanced or recurrent gastric cancer, and assessed the results and safety. One treatment cycle consisted of TS-1, 80 mg/m2/day, for 28 days followed by a 14-day rest period. The efficacy rate in the cases capable of being evaluated was 30.1% (11/36), and 25.0%, (7/28) when TS-1 was used as monotherapy. The efficacy rate was lower than in a phase II study, however, the median survival time (MST) of 10.7 months for the patients as a whole, the 1-year survival rate of 43.2%, and the 2-year survival rate of 20.5% were favorable. There were many NC cases in which long-term therapy was possible, and they contributed to the long-term survival. The incidence of adverse events was 84.1%, but the incidence of grade 3 or more events was low at 13.6%. Since TS-1 is highly efficacious and safe, as well as convenient because of being an oral preparation, it appears that it can be ranked as the drug of first choice for chemotherapy of far advanced or recurrent gastric cancer. PMID- 15751633 TI - [Pilot study of concomitant TS-1/CDGP radiotherapy for esophageal cancer]. AB - Although surgery is treatment of choice for esophageal cancer, radiochemotherapy is being employed throughout Japan for the purpose of improving patient QOL. The results of this therapy are reported to be comparable to those associated with surgical treatment. However, since concomitant 5-FU/CDDP radiotherapy, currently the treatment of choice when implementing radiochemotherapy, is associated with a comparatively high incidence of gastrointestinal disorders and requires continuous intravenous infusion for 24 hours, it lowers the level of patient QOL. We have proposed a clinical study of concomitant TS-1/CDGP radiotherapy for the purpose of maintaining patient QOL and improving outcome. We conducted a pilot study prior to the phase I and II studies. The study was conducted on six cases and favorable results were obtained, consisting of a CR rate of 66.7% and a two year survival rate of 50%. Although bone marrow inhibition was observed as an adverse side effect, gastrointestinal disorders that were discernible to the patients were extremely mild, and patient QOL was able to be maintained. CR was observed in 2 cases who were positive for DPD as determined by immunostaining. We are planning on conducting phase I and II studies in the future based on the potential for this treatment to contribute to the preservation of patient QOL and improve prognosis. PMID- 15751634 TI - [Quality of life and psychological traits of patients undergoing surgery for gastrointestinal cancer]. AB - The purpose of this study was to assess the pre- and post-operative quality of life (QOL) of patients with gastrointestinal cancer and to investigate the relationship between QOL and various psychological and clinical factors. Eighty five patients who underwent surgery for gastrointestinal cancer and 26 control patients undergoing surgery for digestive diseases other than cancer were interviewed. Two tests were administered to assess QOL and psychological status, respectively: the Japanese-language version of the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C 30 and the Japanese-language version of the Hospital Anxiety and Depression Scale (HADS). Each test was administered before surgery, before discharge and 6 months after discharge. Changes in EORTC QLQ-C 30 sub-scale scores over time were compared among advanced phase, early-phase, and control patients. All groups showed significant changes in subscale scores of QOL. The scores of the advanced-phase group indicated worse QOL than the early-phase and control groups in many areas. The results suggest that QOL in gastrointestinal cancer patients is variable over time and is influenced by various clinical factors including the depression and anxiety. PMID- 15751635 TI - [A case of recurrence and systemic metastasis of oral cancer successfully treated with combination chemotherapy of cisplatin and gemcitabine]. AB - We report a case of recurrent and metastatic oral squamous cell carcinoma successfully treated with second-line combination chemotherapy of cisplatin and gemcitabine. A 62-year-old woman, who had previously been treated with surgery and chemoradiation, was referred to our department for systemic metastasis of oral cancer. She had received a combination regimen of cisplatin and 5 fluorouracil against locoregional recurrence. Systemic evaluation revealed metastasis to the lungs, liver and bones. Three cycles of a four-week regimen of combination chemotherapy including cisplatin and gemcitabine resulted in remarkable reduction of the pulmonary and liver metastatic tumors despite frequent neutropenia. Combination chemotherapy of gemcitabine and cisplatin was highly effective in this case, which recurred after a cisplatin-containing regimen. Gemcitabine seems promising and active for head and neck cancer treatment. PMID- 15751636 TI - [Recurrence of esophageal cancer treated by combination TS-1/CDDP therapy]. AB - A 68-year-old man underwent subtotal esophagectomy with two fields lymphadenectomy and postoperative chemotherapy so called low dose FP therapy for advanced esophageal cancer (Stage IIIa, pT 3, pN 1, M 0) in October 1999. As he was diagnosed with a recurrence of esophageal cancer as metastatic lymph node tumors which were placed in the right anterocervical and supraclavicular region in March 2001, he underwent enucleation of metastatic lymph node tumors and postoperative chemoradiation therapy, so-called low-dose FP-R therapy. Recently, since other metastatic lymph node tumors in the neck appeared again in August 2001, he underwent radical neck lymph node dissection and postoperative chemoradiation treatment, so-called FAP-R therapy. In October 2003, a chest CT showed multiple lung tumors. He was diagnosed with multiple metastatic lung tumors originating from esophageal cancer. Then, two courses of a combined chemotherapy consisting of TS-1 and CDDP were administered at an interval of one month. We judged the effect of this chemotherapy to be a partial response (PR), because the largest metastatic lung tumor 18 mm in diameter showed a reduction rate of 81.9%, and other tumors had almost disappeared in the chest CT after the combined therapy. No severe adverse effects of more than grade 3 were observed during this combined therapy. This combined chemotherapy consisting of TS-1 and CDDP may prove effective for treating recurrent cases of esophageal cancer. PMID- 15751637 TI - [A case of a nonresected gastric cancer with peritoneal dissemination maintained on TS-1 and docetaxel combination chemotherapy with good QOL]. AB - In general, treatment for gastric cancer with peritoneal dissemination or recurrent gastric cancer is outside the scope of surgery. The efficacy of new anti-cancer drugs such as TS-1 system was revealed in a controlled study by comparing treatment with non-treatment groups. We performed chemotherapy of TS-1 and docetaxel (TXT) in the outpatient clinic on a 72-year-old nonresected gastric cancer patient accompanied by peritoneal dissemination. Although no killer cell effect was recognized, the patient clinically achieved good QOL by this method for one year and seven months. In conclusion, we reported a treatment method for nonresected gastric cancer, which was treated only on an outpatient basis. The course of this case closely resembled the tumor dormancy therapy performed by Takahashi et al. PMID- 15751638 TI - [A case of liver metastases from cecal cancer successfully treated with fluorouracil and folinic acid (UFT/LV)]. AB - A 76-year-old man had undergone a right hemicolectomy for cecal cancer. Oral UFT (450 mg/day) administration alone was started 2 months following the operation. From a CT scan of the abdomen performed 3 months postoperatively, he was diagnosed with liver metastasis. Because the liver metastasis had progressed, combination oral administration of UFT+LV was started (UFT 450 mg/day, LV 75 mg/day, 4 weeks of therapy followed by a 1-week treatment break). After 1 cycle, a good partial response of that lesion was achieved. The pulmonary metastasis had almost disappeared to within normal limits. In conclusion, this treatment was very safe and effective. PMID- 15751640 TI - [A case of hepatocellular carcinoma with multiple lung metastases responding to intermittent intrahepatic arterial chemotherapy of CDDP+5-FU]. AB - A 72-year-old man who had multiple hepatocellular carcinoma underwent eight times intrahepatic arterial chemotherapies of SMANCS and two times percutaneous ethanol injection (PEIT) therapies over three years, but new diffuse lesions appeared in the liver. He was treated by intermittent intrahepatic arterial chemotherapy with CDDP 10 mg/body/w and 5-FU 500 mg/body/w. Three months after the start of this therapy, the liver tumor was enlarged and multiple lung metastases appeared. But ten months later, the size of the hepatic lesion was reduced and the lung lesion disappeared. Fifteen months later, a solitary metastatic lesion in the left lung was resected. After two years, peritonitis carcinomatosa was observed, and the patient died. PMID- 15751639 TI - [Pharmacokinetic modulating chemotherapy for rectal carcinoma metastases to the liver and lung--a case report]. AB - Pharmacokinetic modulating chemotherapy (PMC) is a new therapeutic regimen for advanced colorectal carcinoma, in which high serum 5-FU concentrations are attained through the inhibition of 5-FU degradation by simultaneously administered uracil. A 67-year-old woman, presented with unresectable multiple hepatic and pulmonary metastases following abdominoperineal resection of rectal carcinoma, was successfully treated by the PMC. The patient was initially treated by 600 mg/m2/day of 5-FU infusion, once a week, and subsequently 5-FU doses were increased to 750 mg/m2/day and then to 1,200 mg/m2/day. Hepatic metastases responded at the dose of 750 mg/m2/day and pulmonary metastases responded at the dose of 1,200 mg/m2/day. The patient remains partial response (>21 months). 5-FU serum concentrations were higher at night time and the peak concentration of 5-FU was obtained at 3 a.m. 5-FU Cmax of 600 mg/m2/day, 750 mg/m2/day and 1,200 mg/m2/day were 254, 329, 531 ng/ml, respectively. The experience of this case, together with literature review, suggests that pulmonary metastases are more resistant to 5-FU than hepatic metastases in patients with colorectal carcinoma. The high serum 5-FU concentrations at night suggest the chronomodulating nature of PMC and are effective for metastatic colorectal carcinoma. PMID- 15751641 TI - [Local recurrence of pancreatic cancer successfully treated with gemcitabine]. AB - We report a patient for whom systemic chemotherapy using gemcitabine was effective against local recurrence of pancreatic cancer. A 58-year-old man underwent pancreatoduodenectomy for a pancreatic head cancer. The diagnosis was Stage IVb poorly-differentiated tubular adenocarcinoma, scirrhous type, pT4, PL (+), P0, H0, pN2. However, after 21 months, gastrointestinal bleeding occurred. Gastroscopy and CT examination revealed a mass at the cut-end of the pancreas invading the stomach. The serum CA19-9 level was found to be elevated. Systemic chemotherapy was performed with a regimen of gemcitabine 1,000 mg/m2/week for 2 weeks, followed by a week rest. The recurrent tumor in the stomach disappeared, and the mass at the cut-end of the pancreas became small. The serum CA 19-9 level regained the normal value. Two years after the diagnosis of recurrence, he returned to work, and his chemotherapy is being continued as an outpatient. PMID- 15751642 TI - [A case of unresectable advanced pancreatic tail cancer successfully treated with gemcitabine hydrochloride]. AB - We report a case of a 76-year-old female with unresectable advanced pancreatic tail cancer in whom systemic chemotherapy using gemcitabine hydrochloride was effective. The patient came to our hospital with a complaint of upper abdominal discomfort and was diagnosed as an advanced pancreatic tail cancer with severe vessel invasion. A curative operation was thought to be impossible, so gemcitabine hydrochloride was administered (800 mg/m2/week x 3/4 weeks). After 2 courses, the CEA, CA19-9 and SPan-1 value were decreased, and reduced tumor size was identified. Sixteen months after the first diagnosis, she died dissemination. PMID- 15751643 TI - [Two-year survivor in response to gemcitabine-based chemotherapy for advanced pancreatic cancer with multiple lung metastases]. AB - A 65-year-old woman was admitted to our hospital for appetite loss and jaundice. Abdominal CT scan and ultrasonography both revealed a tumor in the head of the pancreas. Chest CT scan showed multiple nodules in both lungs. The patient was diagnosed as having advanced pancreatic cancer with multiple lung metastases. The primary tumor showed a partial response to the administration of gemcitabine 1,000 mg/m2 on days 1, 8 and 15 of a 28-day cycle. Concurrent treatment with 5 fluorouracil (200 mg/day) was efficacious against the lung metastases. One year later the pancreatic and lung tumors had enlarged, and cisplatin (20 mg/body) was added to the protocol. For 20 months the patient was treated as an outpatient, maintaining good quality of life. The patient died of progressive disease 25 months after her first hospital admission. PMID- 15751644 TI - [Successful treatment with etoposide by oral administration for recurrence of gastric diffuse large B-cell lymphoma after surgical resection--a case report]. AB - A 64-year-old woman, who had been treated for gastric diffuse large B-cell lymphoma (DLBCL) by total gastrectomy and received 3 courses of CHOP therapy at 61 years of age, was diagnosed with recurrence of DLBCL in the small intestine. After the small intestinal tumor was resected, multiple metastases were found in the liver. Because intensive chemotherapy was difficult for her poor performance status, 50 mg of etoposide daily by oral was administered for 21 consecutive days. After one course of chemotherapy, liver metastases and lymph node swelling almost disappeared without severe adverse effects, and after five courses she achieved complete remission. Though DLBCL invaded the central nervous system, the abdominal regions had been free from recurrence for 12 months. This case report suggests that oral etoposide therapy is useful for gastrointestinal DLBCL which has metastasized to the liver. PMID- 15751645 TI - [A patient with metastatic breast cancer associated with retention of pleural effusion with no response to both CMF and exemestane, whose life was saved by high-dose toremifene]. AB - The patient was a 70-year-old woman, who became aware of a mass in her right breast in 2001, but left it untreated. On March 10, 2003, she visited a nearby doctor with the chief complaint of dyspnea. Since a large painful mass was palpable in the right breast, advanced right breast cancer was suspected, and the patient was referred to our department. Examination revealed the presence of right axillary lymph node metastasis, left pleural effusion, and left atelectasis, and the patient was admitted to our hospital on an emergency basis. Two cycles of CMF were begun on April 2, but CT indicated NC to PD. Therefore, exemestane (EXE, 25 mg/day), was administered on May 13. While the size of the primary lesion was partially decreased, the tumor marker levels were increasing markedly. Administration of EXE was therefore discontinued, and toremifene (TOR, 120 mg/day), was begun. The systemic condition began to improve one month after the start of TOR administration. Two months later, the primary lesion had decreased in size. At after 9 months of TOR treatment, the size of the primary lesion and the tumor marker levels continued to decrease, and both the left pleural effusion and the left atelectasis disappeared. PMID- 15751646 TI - [A case of elderly non-small-cell lung cancer (NSCLC) treated in cooperation with a local hospital and with vinorelbine monotherapy]. AB - We report an elderly advanced NSCLC patient effectively treated with vinorelbine on an outpatient basis in cooperation with his family doctor. A 79-year-old man was referred to our hospital for evaluation of a nodular shadow in his right lung. Chest CT scan showed a 2.0 cm tumor shadow in right S1, and small nodular shadows in right S4 and left S5. Neck lymph node biopsy yielded a diagnosis of adenocarcinoma. The clinical stage was IV, and he underwent chemotherapy of vinorelbine 25 mg/m2 on days 1 and 8. The regimen opted for a setup of a schedule, while the dose of vinorelbine and evaluation by CT were done in our hospital. This regimen was well tolerated and suitable. The patient could receive long-term continuation of the chemotherapy medication, and showed improvement in terms of prolongation of life and QOL. PMID- 15751647 TI - [Acute myocardial infarction induced by lung cancer chemotherapy after radiation of left lung]. AB - A 58-year-old male with non-small cell lung cancer suffered acute myocardial infarction during carboplatin and gemcitabine administration. This case could be cured with percutaneous coronary intervention. The possible underlying mechanisms of infarction are discussed. To our knowledge, this is the first reported case of coronary artery fibrosis demonstrated by intravascular ultrasound during carboplatin-based chemotherapy after radiation of the left lung. PMID- 15751648 TI - [Clinical significance and problems of sentinel node identification for individualization of rectal cancer therapy]. AB - In the surgical treatment of advanced rectal cancer, wide lymphadenectomy has been replaced by autonomic nerve preservation because of serious problems such as postoperative urinary and sexual dysfunction. This may offer a considerable clue to as whether metastasis of lateral lymph node can be diagnosed by a new technique in order to provide more benefits to many patients with advanced rectal cancer. Moreover the sentinel lymph node (SLN) concept has lately attracted attention as a new technique. We studied SLN in 40 patients with advanced lower rectal cancer using 99mTc-Sn colloid as a tracer. SLN was successfully identified in 35 (87.5%) of 40 patients, and the mean number of SLNs identified per patient was 5.6 lymph nodes (range 1-14). In this study overall accuracy was 86%, and it was suggested that lateral lymphadenectomy was excluded in 91%, thanks to application of the SLN technique. Although the role this technique plays in the individualization of the therapy for rectal cancer will be large in the near future, many problems remain to be solved in the exact identification of SLN. PMID- 15751649 TI - [Tumor markers in prostate cancer--clinical significance and future prospect of prostate specific antigen (PSA)]. AB - Prostate specific antigen (PSA), which has high organ specificity, is an excellent tumor marker that has played a significant role in the diagnosis and treatment of prostate cancer. Screening for prostate cancer using PSA is now widely employed in Japan, and increased detection of cases with organ-confined prostate cancer is hoped to result in a decreased number of cancer-specific deaths. Although PSA has also played a critical role in as a marker for staging, assessment of treatment, and recurrence of prostate cancer, many useless biopsies are performed due to its low cancer specificity. To increase the specificity in prostate cancer detection, PSA-related markers including PSAD (PSAPZD), PSAV, and age-specific PSA were advocated, and the ratio of free PSA to total PSA (% free PSA) is also used in a clinical setting. However, since those markers can not satisfactorily exclude benign prostate diseases, various molecular forms of PSA have been analyzed using proteomics and glycomics. Recently, it was demonstrated that plasma free PSA consisted of precursor PSA (pPSA) and other isoforms, suggesting that [-2] pPSA may be a helpful marker for prostate cancer. Our group reported that a sugar chain structure of PSA in the serum of prostate cancer patients is different from that of patients with benign prostate hyperplasia. The different sugar chain structure of PSA can be easily detected by a conventional method and is expected to be useful for differential diagnosis between malignant and benign prostate diseases. PMID- 15751650 TI - [ABO genotyping by duplex amplification and oligonucleotide arrays assay]. AB - OBJECTIVE: ABO genotyping for forensic identification by oligonucleotide chip. METHODS: Oligonucleotide microarrays which could detect 3 different SNPs in exon 6 and exon 7 for ABO genotyping were used. Population studies on ABO was carried out in a sample of 115 unrelated Chinese Han individuals. The method was also applied to cases. RESULTS: The technique could identify 6 genotypes of ABO system. According to the results of population studies, no significant deviations from Hardy-Weinberg equilibrium could be found. The observed and expected heterozygosities were 0.591 and 0.616 respectively. The polymorphic information content was 0.544. The average exclusion probabilities in buos and trios was 0.188 and 0.344 respectively. The discrimination power is 0.777. CONCLUSION: The data and case application demonstrated that ABO typing by oligonucleotide probe arrays was a useful technique for paternity testing and individual identification. PMID- 15751651 TI - [The study on HLA-Cw polymorphism from Xi'an Han population by PCR-sequence specific oligonucleotide probe]. AB - OBJECTIVE: To determine the HLA-Cw allele and genotype frequencies from Xi'an Han population and obtain genetic data. METHODS: The results of HLA-Cw typing for 130 randomly selected from Xi'an Han population were obtained by using the PCR Sequence Specific Oligonucleotide Probes (SSOP). RESULTS: In this investigation, 16 alleles were detected among 130 unrelated individuals with frequencies from 0.0077 to 0.1588 and HLA-Cw*01,03,07 were the most common HLA-Cw alleles. We have made a survey of HLA-Cw alleles frequencies in Xi'an Han Population, with blank frequency being lowered to 0.018 2. CONCLUSION: The distribution of genotype frequencies met the law of Hardy-Weinberg equilibrium by Hi-square test. The frequency data can be used in forensic and paternity tests, transplant matching and anthropology. PMID- 15751652 TI - [Genetic polymorphism of two STR loci D2S1399 and D5S2500 in Eastern Chinese Han population]. AB - OBJECTIVE: To obtain the genetic polymorphism data of two STR loci D2S1399 and D5S2500 in Eastern Chinese Han population. METHODS: Blood samples or buccal swabs of unrelated Han individuals living in eastern China were analyzed using PCR nature polyacrylamide gel electrophoresis-sliver staining method. RESULTS: 11 alleles of D2S1399 and 9 alleles of D5S2500 were observed in the samples respectively, the observed heterozygosity (Ho) values, the discrimination power (DP) values and the power of exclusion (PE) values of D2S1399 and D5S2500 is 0.745 and 0.807, 0.958 and 0.917, 0.554 and 0.643, respectively. CONCLUSION: The result showed that D2S1399 and D5S2500 were highly informative loci and suitable for forensic application. PMID- 15751653 TI - [A sort of meliorative method for testing DNA from sperm smear]. AB - OBJECTIVE: To establish an effective method for testing DNA from the sperm smear. METHODS: We did exploratory research for ninety-one cases of the sperm smear which were accumulated by our laboratory during working hours. Chelex extraction method was used to extract DNA. The digested solution of sperm was purified and concentrated by using Silicon bead. STR loci were typed after PCR amplification by Profile Plus kit. RESULTS: Although there were a few sperm cells on the slide, it is easy to obtain good DNA typing result from it because the amalgamative method we used was high effective for DNA extraction. CONCLUSION: The DNA analysis of sperm smear could offer satisfactory typing results which would be useful in case. PMID- 15751654 TI - [A study of the expression of Fas after fluid percussion brain injury in rats]. AB - OBJECTIVE: To study the pathologic diagnosis and the injury time estimation in light closed encephalon injury. METHODS: Mice were hurt by fluid percussion, and were killed at 15, 30 min, 1, 3 , 6, 12 h, 1, 4, 7, 14 d respectively after injury. The expression of Fas-L in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system. RESULTS: It is showed that the expression of Fas-L could be detected in 1 h after injury, and increased significantly in three hours, and it reached apex 12 h after injury, and decreased gradually four days after injury, and returned normal 14 days after injury. CONCLUSION: This research demonstrated that Fas-L mediated apoptosis appeared not only around brain trauma but also in the brain tissue far away from the traumatic area. It indicted that the expression of Fas-L is a useful target for diagnosis of early brain injury; the regularity of Fas-L expression could be used as one of indication to date the time of brain injury. PMID- 15751655 TI - [Comprehensively analysis the correlation between the height of a person and the length of his/her handprint]. AB - OBJECTIVE: Comprehensively analysis the correlation and the law of variation between the height of a person and the length of his/her handprint. METHODS: Collecting handprint samples of those people of different age and sex from different area all of our country, and adopt the regressive analysis method to study these samples. RESULTS: A sum of useful data and regressive equation were obtained. CONCLUSION: The correlation between the height of a person and the length of his/her handprint is obviously, the approximate height of a person can be reckoned according to the length of his/her handprints. PMID- 15751656 TI - [Preliminary study on the applying value of two measurements for bone age in the cases of minors]. AB - OBJECTIVE: To research the connection of the minor's bone age and chronological age and age for penal responsibility, to discuss the application of Gu's Atlas and Li Guozhen's Percent Numeration to measure the bone age in the cases of immature criminals and the applying value. METHODS: 33 healthy youths, 12-19 years old, were selected, whose left wrists were photographed with X-rays. According to the radiographic appearance of the maturity indicators, the bone age of every person was determined respectively by the Gu's Atlas and Li Guozhen's Percent Numeration, and then compared with his/her chronological age. CMH test and Pearson test were used to evaluate precision and accuracy on concluding the immature age for penal responsibility. RESULTS: Li Guozhen's Percent Numeration was more precise than Gu's Atlas on extrapolating the chronological age of the youth, P<0.005. On extrapolating the age for penal responsibility, the accuracy of the former was 90.91%, and the latter was 78.79%, but the statistic result did not show significant difference (P>0.1). CONCLUSION: Both measurements can apply in determining the criminal cases of immature youths. PMID- 15751657 TI - [Reasons why the quality of medico-legal autopsy in the medical tangle varies from two areas]. AB - OBJECTIVE: To study reasons that the quality of medico-legal autopsy in the medical tangle varies from different area. METHODS: Collecting the cases of medical tangle in two medico-legal agencies, then counting percent of classes on the ten key-points, analyzing the data of the cases by chi-square test and t test. RESULTS: It is indicated that the applied methods and standards of the two agencies are different. There are more different in seven keypoint of medicolegal autopsy by chi-square test. CONCLUSION: Six key-points are found to be more important to medico-legal appraiser, standardization of forensic autopsy, standardization of picking up specimen from the body, diagnosis standardization of the cause of death, consultation system and standardization of writing documents on medico-legal autopsy. PMID- 15751658 TI - [Study of affecting factors of mentally prisoner's competency to serve a sentence]. AB - OBJECTIVE: Explore the affecting factors of mentally prisoner's competency to serve a sentence (CSS), establish the base of quantitative study of CSS. METHODS: Firstly, the researchers compile a questionnaire named legal-psycho ability of competency to serve a sentence questionnaire, then the researchers scaling the object with RTHD, and ask all object complete the questionnaire. there a hypothesis, that the object who are cured in cured ward is incompetent to serve a sentence (ISS), and the other who are stay in rehabilitated ward is competent to serve a sentence (CSS). RESULTS: There are 185 object admitted the study, the ISS group the CSS group have significance between psychiatric and legal aspects. CONCLUSION: At the influence of psychiatric symptoms, the mentally prisoner's competency to serve a sentence had been impaired, and they should been transferred from prison to hospital. PMID- 15751659 TI - [Expressions of NOS isoforms and roles of NO during skin wound healing]. AB - Skin wound healing is an organized process for keeping cutaneous integrity, which needs the complicated interaction between inflammatory cells and biochemical mediators. Nitric oxide synthase (NOS) isoforms, as important factors during skin wound healing, synthesize nitric oxide (NO) which may be involved in the whole event of skin wound healing and play important roles in cell proliferation, differentiation and apoptosis, angiogenesis, matrix deposits and remolding. Furthermore, NO also exerts effects in some cutaneous diseases. In the present paper, the expressions of NOS isoforms and the roles of NO during skin wound healing were reviewed with references to the advances in the studies on skin wound healing. It is suggested that nitric oxide synthases, play significant parts during skin wound healing, which is worthy of further investigation. PMID- 15751660 TI - [The damage of cardiovascular system in heroin abuses]. AB - The heroin abuses can seriously damage human body system, among them the damage of cardiovascular system is various. In this paper those damages involved heart rate, blood pressure, electrocardiogram, heart function, blood circulation, the changes of some material inside, and complications of cardiovascular system are reviewed. PMID- 15751661 TI - [Gap junctions, connexins and sudden death caused by coronary heart disease]. AB - Gap junctions construct hydrophilic trans-membrane channels which adjust the intercellular communication of chemistry and electricity. In the heart, individual cardiac myocytes are linked by gap junctions. These junctions form low resistance pathways along which the electrical impulse flows rapidly and repeatedly between all the myocardium, ensuring their synchronous contraction. In recent years, some researchers have found that connexins, the protein molecules of gap junction channels, are reduced in number or redistributed from intercalated disks (ID) to lateral cell borders in a variety of cardiac disease, especially in ischemic heart disease. The gap junction remodeling is considered to be arrhythmogenic. These findings will lead us to a new realm in the diagnostic of sudden death caused by coronary heart disease. PMID- 15751662 TI - [Application of the age-associated injure in mitochondrial DNA]. AB - Nowadays, the injury in human mitochondrial DNA (mtDNA) is well known to accumulate in various tissues with age. It's significant to further investigate and then apply it to estimation of the age at parenchymas. PMID- 15751663 TI - [Physical injury and mental injury]. AB - Physical and mental injury exist as relative aspect of human injury. They have different medicine and legal feature, as well as mutual dependence and conversion. Physical and mental injury not only present respective character in the classification, but also have difference in specific basis of rating. PMID- 15751664 TI - Effect of perfusionist technique on cerebral embolization during cardiopulmonary bypass. AB - OBJECTIVE: To determine the association between high-intensity transient signals (HITS) and perfusionist interventions, purging techniques, pump flows and venous reservoir blood volume levels during cardiopulmonary bypass. METHODS: Transcranial Doppler was used to detect HITS in the middle cerebral artery during the period of aortic crossclamping in patients undergoing coronary artery bypass grafting. Perfusionist-related interventions were recorded and included blood sampling (including the number of times that the oxygenator sampling manifold was purged), drug bolus injections and infusions (vasopressors, crystalloid and mannitol). Pump flows and venous reservoir volume levels were also documented. RESULTS: There were 534 interventions in 90 patients [median number of interventions per patient: 6 (quartiles: 4, 8)]. The median total HITS count from all interventions was 17 (5, 37). This represented 38% of the total HITS counts during aortic crossclamping. Factors contributing to differences in the HITS count included type of intervention (p <0.0001) and perfusionist (p =0.0012). Blood sampling (p<0.001) and drug bolus injections (p=0.06) had higher HITS counts per patient than infusions. Repetitive purging significantly increased HITS counts (r=0.74; p <0.001). Purging perfusionists (purging: 1-10 times) had higher HITS counts per patient [5 HITS (1, 15) than nonpurgers [0 HITS (0, 1) p <0.0001]. HITS counts were significantly correlated with reservoir volumes (r= 0.20, p=0.017) and pump flow rates (r=0.21, p =0.008). Reservoir volume levels < or =800 mL were associated with higher HITS counts per intervention [11 HITS (2, 27)] during blood sampling compared with higher volume levels [3 HITS (1, 10), p =0.001]. CONCLUSIONS: Cerebral emboli associated with perfusionist interventions can be minimized by not purging the sampling manifold, using continuous infusions rather than bolus injections, and maintaining high blood-volume levels (>800mL) in the venous reservoir. PMID- 15751665 TI - Anticoagulation during extracorporeal circulation under conditions of an ongoing systemic inflammatory response syndrome: effects of heparin. AB - OBJECTIVE: Open-heart surgery with cardiopulmonary bypass (CPB) causes changes in haemostasis. Artificial surfaces are bioincompatible and, thus, may initiate a reaction similar to an acute inflammation. In some patients, this 'postperfusion syndrome' (PPS), which includes changes in haemostasis, is the beginning of a systemic inflammatory response syndrome (SIRS). However, it is not clear whether the changes in coagulation represent a consequence or a main cause of the inflammatory reaction. Thus, the aim of our study was to investigate the cascade of coagulation and the effects of heparin under special circumstances of an ongoing SIRS. METHODS: In a prospective evaluation using standardized operative procedures with CPB, we compared Group A (control group with normal postoperative course, n =20) with Group B (patients with postoperative SIRS, n =12). At six time points beginning before and ending two days after surgery, we measured platelet counts, leucocyte counts and plasma levels of fibrinogen, factor XII and antithrombin III (ATIII), in addition to standard coagulation tests (PTT, TT and ACT). Furthermore, we determined parameters of inflammation, such as C-reactive protein, PCT, IL-6, IL-8, IL-10 and TNF-alpha. RESULTS: In Group B (SIRS), we found a reduced anticoagulation during CPB with significantly lower values for PTT (60+/-7 versus 160+/-11 s), ACT (270+/-33 versus 532+/-44 s) TT (40+/-3 versus 150+/-15 s) compared to the control Group A. Simultaneously, we found a significant increase of factor XII in the SIRS group (191+/-16 versus 10+/-2%). There were no significant differences concerning the preoperative ATIII levels and the intraoperative dosage of heparin; the intraoperative decrease of fibrinogen, ATIII and platelets was comparable in both groups. Furthermore, we could see that significant changes of inflammatory parameters in the SIRS group (increasing levels of TNF-alpha, Il-6, IL-8 and IL-10) occurred at least 30 min after the observed reduction of anticoagulatory effect. CONCLUSIONS: With our results, it could be demonstrated that the development of inflammatory complications after CPB is correlated to a significantly reduced intraoperative effect of heparin. As this reduction of anticoagulation significantly preceded the changes of inflammatory parameters in SIRS patients, we think that a hypercoagulatory state, especially in cases of ongoing inflammation, is an additional trigger of SIRS. PMID- 15751666 TI - Effects of fresh versus old stored blood in the priming solution on whole blood lactate levels during paediatric cardiac surgery. AB - It has been suggested that lactate levels may predict morbidity and mortality in paediatric patients during corrective open-heart surgery. Packed red blood cells (PRBC) are frequently necessary for priming the reservoir used in cardiopulmonary bypass (CPB). The storage of PRBC might cause a significant increase in lactate levels. The purpose of the present study was to quantify the increase in lactate levels in stored red blood cells over time and to compare lactate levels after transfusions of fresh (< or =12 days) versus old blood (>12 days) in 20 patients. We found an increase in lactate levels from 6.0 to 44.7mmol/L (mean 17.0+/-7.8 mmol/L) during storage. Lactate levels were also significantly higher after the onset of CPB in paediatric patients transfused with old blood than in patients transfused with fresh blood (1.43+0.36 versus 3.46+/-0.63, p=0.0006). Our results suggest that the higher lactate levels found after the initiation of CPB should be used with caution when assessing tissue hypoxia and predicting outcome. PMID- 15751667 TI - Clinical benefits of continuous leukocyte filtration during cardiopulmonary bypass in patients undergoing valvular repair or replacement. AB - Valve operations in the form of repair or replacement make up a significant population of patients undergoing surgical procedures in the USA annually with the use of cardiopulmonary bypass. These patients experience a wide range of complications that are considered to be mediated by activation of complement and leukocytes. The extracorporeal perfusion circuit consists of multiple synthetic artificial surfaces. The biocompatibility of the blood contact surfaces is a variable that predisposes patients to an increased risk of complement mediation and activation. This can result in an inflammatory process, causing leukocytes to proliferate and sequester in the major organ systems. The purpose of this study was to determine whether filtration of activated leukocytes improved clinical outcomes following surgical intervention for valve repair or replacement. In this paper, we report a retrospective matched cohort study of 700 patients who underwent valve procedures from June 1999 to December 2002. The control group (CG) consisted of patients who had a conventional arterial line filter. In the study group (SG), patients had a conventional arterial line filter and a leukocyte arterial line filter (Pall Medical, NY). In the SG, blood diverted to the cardioplegia system was also leukocyte depleted to enhance myocardial preservation by adapting this device to the outflow port on the filter. Patient characteristics were similar for the SG and the CG, including 228 males and 122 females, mean age (62.4 versus 64.2 years), cardiopulmonary bypass time (127+/-64 versus 116+/-53 min), and aortic crossclamp time (84+/-23 versus 81+/-23 min). Our results demonstrate that the SG achieved statistically significant reduction in the time to extubation (p =0.03) and the number of patients with prolonged intubation in excess of 24 hours (p <0.04), in addition to improved postoperative oxygenation (p=0.01), and decreased length of hospital stay (p =0.03). We believe that leukocyte filters are clinically beneficial, as demonstrated by the results presented in this study. PMID- 15751668 TI - Prospective study on cardiopulmonary bypass prime reduction and its effect on intraoperative blood product and hemoconcentrator use. AB - PURPOSE: Evaluate the feasibility and clinical significance of crystalloid prime reduction during the initiation of cardiopulmonary bypass (CPB) using a modified bridge on the cardioplegia delivery system. METHODS: Prospective trial of crystalloid prime reduction using a standard Duraflow-coated CPB circuit and Vanguard 2:1 cardio plegia delivery system. Standard prime volume was 1500 cc of Plasmalyte. Prime was reduced via the bridge in the cardioplegia system during initiation of CPB. Packed red blood cells (PRBC) were transfused for hematocrit (Hct) less than 24% while rewarming. A hemoconcentrator was used if the patient's circulating blood volume exceeded 150% of calculated. All data were prospectively collected. RESULTS: Two hundred and twenty-two consecutive patients undergoing cardiac surgery utilizing CPB were evaluated. There were 107 patients with normal prime volume (NPV) and 115 patients with reduced prime volume (RPV). There was no significant difference in sex, mean age, weight, body surface area (BSA), pre-op Hct, procedure time or procedure between the two groups. There was no difference in total crystalloids infused by the anesthetists (average NPV 1205 cc versus RPV 1148 cc). The average RPV was 622 cc (range 400-1100 cc) or a 59% reduction. Post op Hct revealed no difference (NPV 28% versus RPV 29%). There was a 24% reduction in patients requiring PRBC (NPV n=23 versus RPV n=18). The use of hemoconcentrators was reduced by 49% (NPV n=18 versus RPV n =11). The average urine output for both groups exceeded 100 cc/hour while on CPB. CONCLUSION: Using a modified cardioplegia delivery system is a safe and effective method of CPB prime reduction. A RPV resulted in fewer patients requiring PRBC transfusions and fewer hemoconcentrators used. Based on our experience, we would recommend attempting to reduce prime volume in all patients undergoing CPB. PMID- 15751669 TI - Particle separation using ultrasound can be used with human shed mediastinal blood. AB - BACKGROUND: Shed mediastinal blood collected by cardiotomy suction has been shown to be a large contributor to lipid microemboli ending up in different organs. The aim of this study was to test the separation efficiency on human shed blood of a new separation method developed to meet this demand. METHODS: Shed mediastinal blood collected from the pericardial cavity of 13 patients undergoing cardiac surgery with cardiopulmonary bypass was collected. The blood was processed in an eight-channel parallel PARSUS separator, and separation efficiency was determined. RESULTS: Erythrocyte recovery, in terms of a separation ratio, varied between 68% and 91%. Minor electrolyte changes took place, where levels of sodium increased and levels of potassium and calcium decreased. CONCLUSION: This study demonstrates that PARSUS technology can be used on human shed mediastinal blood with good separation efficiency. The technology is, thereby, suggested to have future clinical relevance. PMID- 15751670 TI - The intraoperative effect of pentoxifylline on the inflammatory process and leukocytes in cardiac surgery patients undergoing cardiopulmonary bypass. AB - BACKGROUND: Inflammation plays a pivotal role in the pathogenesis of organ dysfunction after cardiopulmonary bypass (CPB). The aim of this study was to investigate whether pentoxifylline (PTX) has effects on the inflammatory process and leukocytes in cardiac surgery patients undergoing CPB. MATERIAL AND METHODS: A double-blind, prospective, randomized, placebo-controlled study was undertaken to assess the effect of PTX on leukocyte counts, tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and C-reactive protein (CRP) levels in 60 patients undergoing CPB for elective coronary artery bypass grafting. In 30 patients, 200 mg of PTX was added to 500 mL NaCl and perfused for 180 min after induction of anaesthesia and also 100 mg of PTX was added to the warm cardioplegic solution; another 30 patients received saline solution as placebo. RESULTS: All measurements were performed before PTX infusion (T0), after induction of anaesthesia (T1), 30 min after weaning from CPB (T2), and 6 hours (T3) and 24 hours postoperatively (T4). PTX did not change the percentage of eosinophils, basophils, neutrophils, monocytes, or lymphocytes, or CRP levels. In the control group, however, total leukocyte count and IL-6 level at T3 and T4 period were significantly higher than the study group. The progressive increment in TNF-alpha level observed at each period was also significantly prominent in the control group. CONCLUSION: CPB-related whole body inflammatory response could be partially inhibited by intraoperative PTX administration. This effect of PTX would be helpful in preventing the well-known complications of CPB-induced systemic inflammation. PMID- 15751671 TI - Renal medullary hypoxia during experimental cardiopulmonary bypass: a pilot study. AB - OBJECTIVES: To determine the effect of cardiopulmonary bypass (CPB) on renal medullary oxygenation. DESIGN: Observational. SETTING: Laboratory. PARTICIPANTS: Pigs (n=3). INTERVENTIONS: Following induction of general anesthesia, a Paratrend blood gas probe was placed directly into the left renal medulla. Two animals were subjected to 90 min of CPB, while a third served as a non-CPB control. A probe was also placed in the left renal pelvis of one (CPB) animal to allow direct urine PO2 measurements. MEASUREMENTS AND MAIN RESULTS: Medullary hypoxia (PO2 <65 mmHg) was evident prior to CPB. With the onset of CPB, medullary PO2 further declined to nearly unmeasurable levels; PCO2 and pH were unchanged. Brief circulatory arrest during CPB in one animal resulted in rapid additional PCO2 rise and pH decline that corrected with reperfusion. Following the cessation of CPB, medullary PO2 gradually increased, but remained lower than pre-CPB levels. No changes in medullary PO2 were observed in the sham animal. Renal pelvis urine PO2, but not pH or PCO2, appeared to correlate with medullary values at all times. CONCLUSIONS: Our findings indicate that renal medullary hypoxia is extreme during CPB and may persist following CPB. These data suggest a basis for the vulnerability of the kidney to injury during cardiac surgery. Renal pelvis urine PO2 appears to correlate closely with medullary PO2 and may be a useful tool for studying medullary oxygenation during CPB in humans. PMID- 15751672 TI - Measurements of detector nonlinearity at 193 nm. AB - We have developed a measurement system based on a correlation method to characterize the nonlinearity of a detector's response over a large range of laser pulse energy. The system consists of an excimer-laser source, beam-shaping optics, a beam splitter, a monitor detector, a set of optical filters, and the detector under test. Detector nonlinearities as large as 10% or greater over an entire measurement range at an excimer-laser wavelength of 193 nm are observed. The measurement range of the current system is approximately 300 nJ to 50 mJ of laser pulse energy at the detector under test. The typical expanded measurement uncertainty of nonlinearity is 0.6% (k = 2). PMID- 15751673 TI - Compact description of substrate-related aberrations in high numerical-aperture optical disk readout. AB - Optical disks are read out by focusing a beam of high numerical aperture (NA) through the substrate. Deviations of the thickness from the nominal value result in spherical aberration; tilting the substrate results in coma. Exact analytical expressions for the rms aberration per micrometer thickness mismatch (for spherical aberration) and per degree tilt (for coma) are derived. The paraxial estimates for these sensitivities proportional to NA4 (spherical aberration) and NA3 (coma) underestimate the exact values by a factor of approximately 2 for the value NA = 0.85, corresponding to the new Blu-ray disk format. Expansion of the aberration function in Zernike aberrations shows that the exact aberration functions are well described by the lowest-order Zernike spherical aberration (A40) and coma (A31) term for all but the very highest NA values. PMID- 15751674 TI - Uniform large-area x-ray imaging at 9 keV using a backlit pinhole. AB - The development and application of point backlighting at high x-ray energies is an essential step in diagnosing radiation-driven experiments. The point backlighting technique provides uniform backlighter irradiance over a large field of view. This technique circumvents the large laser energy required for area backlighters at energies of 9 keV and above. We present the results of a Zn 9 keV point-backlighter source using the technique of pinhole aperturing to define the source size and hence the resolution. Details of the design and application of this technique to an undriven gold-walled hohlraum are described. PMID- 15751675 TI - Simple method for accurate characterization of birefringent crystals. AB - We present a simple method to determine the cutting angle and thickness of birefringent crystals. Our method is based on chromatic polarization interferometry and allows for accuracies of typically 0.1 degrees in the cutting angle and 0.5% in the thickness. PMID- 15751676 TI - ACR II: improved absolute cryogenic radiometer for low background infrared calibrations. AB - A second-generation absolute cryogenic radiometer (ACR II) was developed for use at the Low Background Infrared calibration facility at the National Institute of Standards and Technology. The need for spectral calibrations of very sensitive [D* = 10(14) cm (Hz)1/2W(-1)] infrared detectors necessitated the use of a cryogenic infrared monochromator and a more sensitive radiometer. The improved low-power performance of the ACR II compared with the older absolute cryogenic radiometer (ACR) has also made it useful as the primary standard for the calibration of cryogenic blackbody sources that are used as low-power infrared sources. The responsivity of the new radiometer's receiver is 210 K/mW with a type A (random component) standard uncertainty of at most 7 pW when making power measurements of less than 10 nW. The original ACR has a responsivity of 29 K/mW and has a type A standard uncertainty of approximately 100 pW when making a similar low-noise-power measurement. Other properties of the radiometers are also described and compared. PMID- 15751677 TI - Hybrid radiative-transfer-diffusion model for optical tomography. AB - A hybrid radiative-transfer-diffusion model for optical tomography is proposed. The light propagation is modeled with the radiative-transfer equation in the vicinity of the laser sources, and the diffusion approximation is used elsewhere in the domain. The solution of the radiative-transfer equation is used to construct a Dirichlet boundary condition for the diffusion approximation on a fictitious interface within the object. This boundary condition constitutes an approximative distributed source model for the diffusion approximation in the remaining area. The results from the proposed approach are compared with finite element solutions of the radiative-transfer equation and the diffusion approximation and Monte Carlo simulation. The results show that the method improves the accuracy of the forward model compared with the conventional diffusion model. PMID- 15751678 TI - Spatially selective laser irradiation method controlled by real-time image analysis: optical aspects. AB - We present a spatially selective irradiation method for laser treatment of biological surfaces. The purpose is to irradiate only the pathological targets and to preserve the healthy surrounding parts. We are interested here in the optical arrangement of the device and in the evaluation of its physical limitations before it is used for medical purposes. The method uses real-time image analysis applied to the video imagery of the surface to be treated. The result of the image analysis generates the control signal to a spatial light modulator, allowing us to project an adequate repartition of laser irradiance on the surface to be treated. Experimental results demonstrate a spatial selectivity of approximately 10 microm for a 6.7 mm x 5 mm field. The optical system has no moving parts (unlike galvanometric scanners) and is able to irradiate multiple targets simultaneously with different doses. PMID- 15751679 TI - Prism-based excitation wavelength selection for multicolor fluorescence coincidence measurements. AB - We have designed and constructed a prism setup for multiple-color confocal fluorescence experiments. The prism setup permits easy selection of any color or any combination of colors from a multicolor light source, such as a mixed-gas argon-krypton-ion laser. The selected colors emerging from the prism setup are, by design, optimally overlapped in the focus of a high-numerical-aperture objective, such as that commonly used in single-molecule fluorescence experiments. The various excitation powers can be easily adjusted in this setup. We will exemplify the potential of this setup in single-molecule fluorescence emission spectroscopy. PMID- 15751680 TI - Luminance addition of a stack of multidomain liquid-crystal displays and capability for depth-fused three-dimensional display application. AB - A stack of liquid-crystal displays is expected to reduce visual fatigue caused by a flat-panel three-dimensional (3D) display. We previously developed a compact depth-fused 3D (DFD) display by using a stack of two twisted-nematic (liquid crystal) LC panels, but its viewing-angle characteristics and color reproducibility were not so good. Therefore recent wide-view LC panels should be used. We report calculated and measured luminance addition characteristics, which are essential for evaluating 3D display characteristics, of a stack of two multidomain LC panels. We found that LC panels with super in-plane switching, patterned vertical alignment, multiple vertical alignment, and continuous pinwheel alignment modes are suitable for DFD display application. PMID- 15751681 TI - Low-refractive-index dye-aggregate films with small absorption based on anomalous dispersion. AB - Complex-refractive-index spectra of Squarylium (SQ) dye-aggregate films deposited upon metal films have been investigated by measurements of properties of the films including absorption spectra (AS) and attenuated total reflection. Complex refractive indices are estimated by Kramers-Kronig analysis for the AS and by a theoretical curve-fitting analysis for attenuated total reflection. The dye aggregate films exhibited an absorption that was blueshifted from that of a monomer, as a result of the H-aggregate formation of SQ molecules, and had a changing refractive index with anomalous dispersion about the H-absorption band. From both measurements of the SQ films it was found that there is a region of low absorption in the short-wavelength side of the absorption band and that the refractive index there is lower than that of glass. PMID- 15751682 TI - Optically monitored dip coating as a contactless viscometry method for liquid films. AB - Real-time interferometric monitoring of the dip coating process is applied to the study of properties of flowing liquids. Nonvolatile Newtonian oils are considered, allowing validity of a simple model after the steady state is reached where film physical thickness depends on time as t(-1/2). Measurement of two distinct mineral oil standards, under several withdrawing speeds, resulted in kinematic viscosities of 1.17+/-0.03 and 9.9+/-0.2 S (1S = 1 cm2/s). Agreement of these results with nominal values from the manufacturer suggests that interferometric monitoring of dip coating may become a valuable method for accurate, contactless viscometry of liquid films. Advantages and present limitations are discussed. PMID- 15751683 TI - X-ray telescope onboard Astro-E. III. Guidelines to performance improvements and optimization of the ray-tracing simulator. AB - We present a detailed study of the performance of the Astro-E x-ray telescope (XRT) onboard the Astro-E satellite. As described in preceding papers the ground based calibrations of the Astro-E XRT revealed that its image quality and effective area are somewhat worse than that expected from the original design. Conceivable causes for such performance degradation are examined by x-ray and optical microscopic measurements at various levels, such as individual reflectors, sectors, and quadrants of the XRT and their alignments. We can attribute, based on detailed measurements, the degradation of the image quality to a slope error in the individual reflectors and the positioning error of reflectors. As for the deficit of the effective area, the shadowing of x rays within the XRT body is the dominant factor. Error budgets for the performance degradation of the Astro-E XRT are summarized. The ray-tracing simulator, which is needed to construct the response function for arbitrary off-axis angles and spatial distributions of any celestial x-ray sources, has been developed and tuned based on the results of detailed measurements. The ray-tracing simulation provides results that are consistent within 3% with the real measurement except for large off-axis angles and higher energies. We propose, based on knowledge obtained from all the measurements and simulations, several plans for future developments to improve the performance of the nested thin-foil mirrors. PMID- 15751685 TI - Two-dimensional phase-measuring profilometry. AB - An improved method is proposed to perform contouring of an object based on phase measuring profilometry with a grid grating. Two phases unwrapped in perpendicular directions are obtained with the help of an adaptive bandpass filter and are used to unwrap the phases and to appoint the edge points of a shaded area and the object area. The height distribution of the object is obtained with the geometric relationship between coordinates and phases. The main axis of the projector and the main axis of the camera are not crossed and are also not in the same plane in order to arrange a measuring system easily and conveniently. The experimental results show that this technique is available for practical applications. PMID- 15751684 TI - Spatial deconvolution technique to improve the accuracy of reconstructed three dimensional diffuse optical tomographic images. AB - A straightforward spatial deconvolution operation is presented that seeks to invert the information-blurring property of first-order perturbation algorithms for diffuse optical tomography (DOT) image reconstruction. The method that was developed to generate these deconvolving operators, or filters, was conceptually based on the frequency-encoding process used in magnetic resonance imaging. The computation of an image-correcting filter involves the solution of a large system of linear equations, in which known true distributions and the corresponding recovered distributions are compared. Conversely, application of a filter involves only a simple matrix multiplication. Simulation results show that application of this deconvolution operation to three-dimensional DOT images reconstructed by the solution of a first-order perturbation equation (Born approximation) can yield marked enhancement of image quality. In the examples considered, use of image-correcting filters produces obvious improvements in image quality, in terms of both location and mirco(a) of the inclusions. The displacements between the true and recovered locations of an inclusion's centroid location are as small as 1 mm, in an 8-cm-diameter medium with 1.5-cm-diameter inclusions, and the peak value of the recovered micro(a) for the inclusions deviates from the true value by as little as 5%. PMID- 15751686 TI - Wavelet analysis of speckle patterns with a temporal carrier. AB - A novel temporal phase-analysis technique that is based on wavelet analysis and a temporal carrier is presented. To measure displacement on a vibrating object by using electronic speckle pattern interferometry, one captures a series of speckle patterns, using a high-speed CCD camera. To avoid ambiguity in phase estimation, a temporal carrier is generated by a piezoelectric transducer stage in the reference beam of the interferometer. The intensity variation of each pixel on recorded images is then analyzed along the time axis by a robust mathematical tool, i.e., a complex Morlet wavelet transform. After the temporal carrier is removed, the absolute displacement of a vibrating object is obtained without the need for temporal or spatial phase unwrapping. The results obtained by a wavelet transform are compared with those from a temporal Fourier transform. PMID- 15751687 TI - Comprehensive assessment of the two-flow model's shape factors in aquatic environments. AB - The applicability of the two-flow model to problems is dependent on assumptions that are used to reduce the model to two equations with two unknowns. Such a reduction requires proper parameterization of the shape factors. An extensive data set that reveals the relationships of shape factors to scattering-to absorption ratios (0.1 < or = b/a < or = 30) and to the mean cosine of scattering (0.66 < or = micro(s) < or = 0.95) as obtained with a Monte Carlo model is presented. The sensitivity of the computed irradiance reflectance to the shape factors is also addressed. Results show increasing sensitivity of irradiance reflectance to the shape factors with increasing scattering-to-absorption ratio and with increasing anisotropy of the scattering phase function. For waters with highly anisotropic scattering phase functions, high scattering-to-absorption ratios, or both, the proper parameterization of the shape factors must be considered. A lookup table of the shape factors' depth-dependent values for various optical conditions is also presented. PMID- 15751688 TI - Average intensity and spreading of cosh-Gaussian laser beams in the turbulent atmosphere. AB - The average intensity and spreading of cosh-Gaussian laser beams in the turbulent atmosphere are examined. Our research is based principally on formulating the average-intensity profile at the receiver plane for cosh-Gaussian excitation. The limiting cases of our formulation for the average intensity are found to reduce correctly to the existing Gaussian beam wave result in turbulence and the cosh Gaussian beam result in free space (in the absence of turbulence). The average intensity and the broadening of the cosh-Gaussian beam wave after it propagates in the turbulent atmosphere are numerically evaluated versus source size, beam displacement, link length, structure constant, and two wavelengths of 0.85 and 1.55 microm, which are most widely used in currently employed free-space-optical links. Results indicate that in turbulence the beam is widened beyond its free space diffraction values. At the receiver plane, analogous to the case of free space, this diffraction eventually leads to transformation of the cosh-Gaussian beam into an oscillatory average-intensity profile with a Gaussian envelope. PMID- 15751689 TI - Second-generation laser firefly clusters: improved scheme for distributed sensing in the atmosphere. AB - We present an improved concept of laser firefly clustering for atmospheric probing, elaborating on previous published work. The laser firefly cluster is a mobile, flexible, and versatile distributed-sensing system, whose purpose is to profile the chemical and the particulate composition of the atmosphere for pollution monitoring, meteorology, detection of contamination, etc. The fireflies are deployed in situ at the altitude of interest and evoke a backscatter response from aerosols and molecules in the immediate vicinity by using a coded laser signal. The enhanced, second-generation system affords better performance at lower energy cost and has a considerably increased scope of application. A numerical example demonstrates the potential of the innovative system. PMID- 15751690 TI - Closed-loop stability and performance analysis of least-squares and minimum variance control algorithms for multiconjugate adaptive optics. AB - Recent progress has been made to compute efficiently the open-loop minimum variance reconstructor (MVR) for multiconjugate adaptive optics systems by a combination of sparse matrix and iterative techniques. Using spectral analysis, I show that a closed-loop laser guide star multiconjugate adaptive optics control algorithm consisting of MVR cascaded with an integrator control law is unstable. Tosolve this problem, a computationally efficient pseudo-open-loop control (POLC) method was recently proposed. I give a theoretical proof of the stability of this method and demonstrate its superior performance and robustness against misregistration errors compared with conventional least-squares control. This can be accounted for by the fact that POLC incorporates turbulence statistics through its regularization term that can be interpreted as spatial filtering, yielding increased robustness to misregistration. For the Gemini-South 8-m telescope multiconjugate system and for median Cerro Pachon seeing, the performance of POLC in terms of rms wave-front error averaged over a 1-arc min field of view is approximately three times superior to that of a least-squares reconstructor. Performance degradation due to 30% translational misregistration on all three mirrors is approximately a 30% increased rms wave-front error, whereas a least squares reconstructor is unstable at such a misregistration level. PMID- 15751691 TI - Robustness study of the pseudo open-loop controller for multiconjugate adaptive optics. AB - Robustness of the recently proposed "pseudo open-loop control" algorithm against various system errors has been investigated for the representative example of the Gemini-South 8-m telescope multiconjugate adaptive-optics system. The existing model to represent the adaptive-optics system with pseudo open-loop control has been modified to account for misalignments, noise and calibration errors in deformable mirrors, and wave-front sensors. Comparison with the conventional least-squares control model has been done. We show with the aid of both transfer function pole-placement analysis and Monte Carlo simulations that POLC remains remarkably stable and robust against very large levels of system errors and outperforms in this respect least-squares control. Approximate stability margins as well as performance metrics such as Strehl ratios and rms wave-front residuals averaged over a 1-arc min field of view have been computed for different types and levels of system errors to quantify the expected performance degradation. PMID- 15751692 TI - Kinetic behavior of polymer-coated long-period-grating fiber-optic sensors. AB - A new method of analysis employing the time-dependent response of long-period grating (LPG) fiber-optic sensors is introduced. The current kinetic approach allows analysis of the time-dependent wavelength shift of the sensor, in contrast to previous studies, in which the LPG sensing element has been operated in an equilibrium mode and modeled with Langmuir adsorption behavior. A detailed kinetic model presented is based on diffusion of the analyte through the outer protective membrane coating into the affinity coating, which is bound to the fiber cladding. A simpler phenomenological approach presented is based on measurement of the slope of the time-dependent response of the LPG sensor. We demonstrate the principles of the kinetic methods by employing a commercial Cu+2 sensor with a carboxymethylcellulose sensing element. The detailed mathematical model fits the time-dependent behavior well and provides a means of calibrating the concentration-dependent time response. In the current approach, copper concentrations below parts per 10(6) are reliably analyzed. The kinetic model allows early-time measurement for low concentrations of the analyte, where equilibration times are long. This kinetic model should be generally applicable to other affinity-coated LPG fiber-optic sensors. PMID- 15751693 TI - Numerical simulation of a high-average-power diode-pumped ytterbium-doped YAG laser with an unstable cavity and a super-Gaussian mirror. AB - A numerical technique with which to compute the output characteristics of a solid state laser with an unstable cavity and a super-Gaussian coupling mirror is proposed. This technique is applied to an Yb:YAG actively Q-switched laser. With this formalism, the mode formation for the fundamental mode is analyzed and the performance achievable by such a laser for various cavity parameters is determined. Then the results obtained with such a cavity are compared with those given for a stable cavity with graded phase output mirror that is also used for obtaining super-Gaussian mode. PMID- 15751694 TI - Direct quasi-phase-matched fourth-harmonic generation. AB - We have theoretically demonstrated direct fourth-harmonic generation (FHG) based on a quasi-phase-matching (QPM) configuration in periodically poled lithium niobate. The wavelength dependence of period of FHG QPM gratings is calculated. Bandwidths of fundamental wavelength, temperature, and incident angle are also studied. We find a very wide bandwidth, as large as 115 nm, of fundamental wavelength near the wavelength of 3797 nm with the QPM period of 9.73 microm. The numerical calculation shows that the conversion efficiency for QPM FHG and cascading QPM by two-step second-harmonic generation is almost identical. PMID- 15751696 TI - Computation of the modal reflectivity for a partially etched mirror: application for integration of a laser diode and a waveguide. AB - We demonstrate a fast, accurate for weak waveguiding, approximate solution to the problem of computing the modal reflection and transmission coefficients at a discontinuity between two asymmetric multilayer waveguides together with a rigorous solution to check the validity of the approximate but fast approach. The proposed algorithm is analogous to the widely used approximation proposed by Vassallo [Electron. Lett. 21, 333 (1985)] for the case when the medium after discontinuity is uniform and has a large applicability potential for optoelectronic applications. As an example, the case of optoelectronic integration between an active laser diode and a passive waveguide device is analyzed. PMID- 15751695 TI - Numerical modeling of polarization conversion in semiconductor electro-optic modulators. AB - An accurate numerical simulation study of a polarization conversion phenomenon in deeply etched semiconductor electro-optic waveguide modulators is presented. Based on a powerful and versatile finite element package, the effect of various imperfect fabrication conditions on unwanted and unexpected polarization conversion in electro-optic semiconductor modulators is, for the first time to our knowledge reported and explained in terms of its origin. PMID- 15751697 TI - Influence of atmospheric and system parameters on multiple scattering in spaceborne backscatter lidar measurements. AB - We have simulated backscatter signals of spaceborne lidar systems with the help of a Monte Carlo model. Calculations were performed for various combinations of system parameters. As typical examples of atmospheric observation targets, two kinds of cirrus cloud and two kinds of aerosol were considered. Both total multiple scattering and the significance of individual higher scattering orders are discussed. For all cases, an approximate multiple scattering factor F was calculated that can be used to correct the single-scattering lidar equation to account also for multiple scattering. PMID- 15751698 TI - First retrieval of vertical profiles of turbulence characteristics and horizontal wind velocity from solar transmission measurements at 212 and 405 GHz. AB - We report on the investigation and successful application of vertical profiling of the structure parameter C2m and of the outer scale L0 of absorption fluctuations and of the horizontal wind velocity (vector) during daytime by the analysis of solar transmission measurements. The method is relatively simple and straightforward so that the presented (or a similar) technique could be used in the routine remote sensing of daytime C2m, L0, and wind profiles. It requires multiple beams pointing in different directions at the Sun. The retrieved profiles are consistent with the current knowledge of atmospheric physics. Simultaneous in situ wind velocity measurements agree with the retrieved wind velocity in the lowest 100 m above ground within the measurement uncertainties of less than +/-2 m/s. The derived values of C2m at 200 m above ground are in good agreement (within a factor of 1.5) with the findings of an earlier investigation at the same test site. Finally, it is shown that irradiance fluctuations of millimeter and submillimeter waves are dominantly affected by humidity fluctuations, even at a dry and elevated site. PMID- 15751699 TI - Antinoise approximation of the lidar signal with wavelet neural networks. AB - We propose a new, to our knowledge, denoising method for lidar signals based on a regression model and a wavelet neural network (WNN) that permits the regression model not only to have a good wavelet approximation property but also to make a neural network that has a self-learning and adaptive capability for increasing the quality of lidar signals. Specifically, we investigate the performance of the WNN for antinoise approximation of lidar signals by simultaneously addressing simulated and real lidar signals. To clarify the antinoise approximation capability of the WNN for lidar signals, we calculate the atmosphere temperature profile with the real signal processed by the WNN. To show the contrast, we also demonstrate the results of the Monte Carlo moving average method and the finite impulse response filter. Finally, the experimental results show that our proposed approach is significantly superior to the traditional methods. PMID- 15751700 TI - Trace detection of explosives with low vapor emissions by laser surface photofragmentation-fragment detection spectroscopy with an improved ionization probe. AB - Trace explosive residues are measured in real time by surface laser photofragmentation-fragment detection (SPF-FD) spectroscopy at ambient conditions. A 248-nm laser photofragments the target residue on a substrate, and a 226-nm laser ionizes the resulting NO fragment by resonance-enhanced multiphoton ionization by means of its A-X (0, 0) transitions near 226 nm. We tested two probes on selected explosives and modeled their electric field in the presence of a substrate with an ion optics simulation program. The limits of detection range from 1 to 15 ng/cm2 (signal-to-noise ratio of 3) at 1 atm and 298 K and depend on the electrode orientation and mechanism for NO formation. PMID- 15751701 TI - Influence of spatiotemporal coupling induced by an ultrashort laser pulse shaper on a focused beam profile. AB - 4f pulse shapers have been widely used to temporally manipulate femtosecond optical pulses by spectral filtering. When the temporal waveform is manipulated with a spatial light modulator consisting of segmented pixels, the spatial profile of the output beam also varies because of diffraction at the pixel array, which is known as a spatiotemporal coupling effect. This effect produces a complicated spatio-temporal profile near the focus of the ultrashort pulses, which may affect the interpretation of experimental results obtained with shaped ultrashort pulses. We investigate the spatial intensity distribution at the focus of temporally shaped pulses through ablation experiments. The three-dimensional space-time beam profile is also numerically calculated. PMID- 15751702 TI - [What should we understand by tuberculosis cure?]. PMID- 15751703 TI - [On the treatment of bronchial obstruction in patients with respiratory tuberculosis]. PMID- 15751704 TI - [Organization of antituberculous care in penitentiary facilities]. AB - The paper presents the organizational aspects of antituberculous care delivered at penitentiary institutions of the Russian Federation. The priorities of development, including the interaction with civil public health care of Russia and the continuation of tuberculosis-controlling activities jointly with international organizations, are identified. PMID- 15751705 TI - [Drug-resistant tuberculosis in villagers]. PMID- 15751706 TI - [Mini-invasive surgery for complications due to pulmonary tuberculosis]. PMID- 15751707 TI - [Vibration massage in the prevention of postresection complications and in the clinical rehabilitation of patients with pulmonary tuberculosis after surgical interventions]. AB - A new procedure has been developed for prevention of early postresectional complications and for clinical rehabilitation of patients with tuberculosis after surgical interventions. The procedure is based on a 13-14-day course of regional and systemic physiotherapy applied to a patient's respiratory organs and chest after surgery in a certain order with the use of tuberculostatic agents. Regional physiotherapy includes local massage of the root of the contralateral operated lung with a light vibromassage apparatus; systemic physiotherapy involves electric vibroacupressure of the whole circumference of the chest with a special device. Early postresection complications were significantly less frequently observed in the study group (n = 60) than in the control group (n = 50) (p < 0.01). The procedure reduces the likelihood of development of a number of pleuropulmonary events (atelectasis, non-specific pneumonia, residual postresection pleural cavity, bronchial fistulas) and promotes functional rehabilitation in patients. PMID- 15751708 TI - [Study of collections of Mycobacterium non-tuberculosis by a restriction test of the amplified fragment spacer sequence 16S-23S of ribosomal DNA]. AB - Fifty mycobacterial cultures were studied by restriction analysis of the amplified fragment of spacer sequence 16S-23S of ribosomal DNA. The following types were identified: M. tuberculosis complex, M. avium, M. intracellulare, M. scrofulaceum, M. kansasii, M. gordonae, M. ulcerans, M. fortuitum, M. smegmatis. The genetic heterogeneity of M. fortuitum was detected. The strains under study belong to 5 patterns: M. fortuitum I, M. fortuitum II, M. fortuitum X, M. fortuitum Y, M. fortuitum Z. The pattern characteristics for M. ulcerans (the size of a CPR product was 370 pn; the characteristics of the enzyme HaeIII induced restriction profile was 214/155 pn) were obtained. PMID- 15751709 TI - [Antibodies to mycobacterial antigens in patients with pulmonary tuberculosis]. AB - The levels of IgG and IgM antibodies to different mycobacterial antigens were studied in patients with varying pulmonary tuberculosis. The higher levels of IgG and IgM to the used antibodies were simultaneously found in patients with infiltrative tuberculosis as compared with healthy individuals. As compared to patients with fibrocavernous pulmonary tuberculosis, those with infiltrative tuberculosis had significantly lower levels of IgG antibodies to BGC sonicate and protein with a molecular weight of 40-70 kD isolated from the strain H37Rv. A correlation was found between the IgM antibodies to protein isolated from the strain H37RV and to H37RV sonicate and the degree of the activity unique to fibrocavernous pulmonary tuberculosis. PMID- 15751710 TI - [The activity of alpha2-macroglobulin and its forms in patients with destructive pulmonary tuberculosis]. AB - A hundred and three patients with destructive pulmonary tuberculosis were examined. These patients had heterodirectional changes in the serum levels of alpha2-macroglobulin (MG) and its forms. The lowering of the level of the F-form of the inhibitor outstripped the decrease in the activity of total alpha2-MG and it was detectable in the lobar processes with moderate symptoms of intoxication and with the inoculation of adjacent segments. The preservation of the low total activity of the inhibitor and its forms during antituberculous therapy characterize a diminished process. The high content of the S form of alpha2-MG is a feature in patients showing positive changes before and 2 months after treatment whereas the changes in total alpha2-MG and its F form may vary. Whether it is expedient to study the fractional composition of alpha2-MG is determined by the heterodirectionality of changes in its S and F forms depending on the pattern of a process. PMID- 15751711 TI - [Interferon-gamma and IgG antibodies to M. tuberculosis in the serum of patients with active pulmonary tuberculosis]. AB - The above parameters in the sera from patients with active tuberculosis (n = 20) and healthy individuals (n = 16) were studied in parallel to examine the relationship and mutual effects of interferon-gamma (IFN-gamma) and IgG antibodies to Mycobacterium tuberculosis, as well as their relationship with some clinical and immunological features of tuberculous infection. The relative and absolute counts of CD4 and CD16 cells were additionally measured in these patients' peripheral blood. Patients with active tuberculosis were found to have significantly elevated serum levels of IFN-gamma, IgG antibodies and CD16 cells with the decreased count of CD4 cells as compared with the similar parameters in healthy individuals. There was no relationship of the serum level of IFN-gamma to the clinical features of tuberculosis. The serum level of IFN-gamma is likely to determine the interferonogenic activity of the causative agent itself. PMID- 15751712 TI - [Diagnostic features of newly diagnosed tuberculosis on different accounts of consulting with a physician]. AB - The specific features of diagnosis were studied in 209 patients with tuberculosis at 9 phthisiological stations of the Yaroslavl Region in 1998-2002. In 96.2% of the patients, tuberculosis was found to have been newly diagnosed in general polyclinics and polyclinics. Only in 3.8% of the patients, the diagnosis of tuberculosis was made at prophylactic fluorography performed on the patients' initiative. A retrospective assessment of outpatient records indicated a significant delay in establishing tuberculosis, by using adequate diagnostic criteria if a physician had professional skills. The authors arrive at the conclusion that dispensary should assume the functions of phthisiopulmonological centers that will secure the continuity of early diagnosis of tuberculosis and its early treatment. PMID- 15751713 TI - [Leukinferon immunomodulation in pulmonary tuberculosis]. PMID- 15751714 TI - [Complex method for examination and management of patients with respiratory tuberculosis during pregnancy, labor, and puerperium]. PMID- 15751715 TI - [Order No 50 "On putting into effect accounting and reporting documents for tuberculosis monitoring" issued by the Ministry of Health on February 13, 2004]. PMID- 15751716 TI - [Reconstruction of soft tissue defects of the fingers using homodigital V-Y flap]. AB - A series of 54 patients is presented in which full-thickness soft-tissue defects on 57 digits were reconstructed using homodigital V-Y flaps. This is a modification of the Moberg procedure, which was designed for coverage of injuries of distal thumb. The V-Y flap is pedicled on two digital neurovascular bundles, possible advancement is up to 2 cm, and V-shaped base of the flap allows direct closure of the proximal defect, without skin grafting. This technique was used for the reconstruction both volar and dorsal tissue defects of the fingers. All flaps healed within 2-4 weeks. 14 patients (15 fingers) were evaluated after they recovered. In all affected fingers active range of motion was satisfactory, only with slight defect of extension in 2 cases. However, sensation of the light touch was decreased in 10 fingers, and 2PD discrimination was abnormal in 5 fingers. The versatility of V-Y technique in various clinical occasions and its low risk of complications was emphasized. This method is very useful, easy to learn even for trainees unfamiliar with microsurgery. PMID- 15751717 TI - [Reconstruction of humeral head retroversion in the treatment of anterior shoulder instability]. AB - Reduction of the physiologic retroversion of the humeral head can lead to shoulder luxation. After shoulder luxation it can also lead to shoulder instability. The aim of the paper was to check the level of humeral head retroversion in patients with anterior shoulder instability and to estimate the effectiveness of treatment of this instability with the derotational Weber osteotomy. The study included 38 patients, 34 males and 4 females, ranged 16-48 yr. (mean 31) treated in our Clinic. Every patient was done CT in order to estimate the level of humeral head retroversion. Patients with reduced humeral head retroversion qualified for operational treatment with Weber method derotational osteotomy. The outcomes were estimated according to the Rowe and Zarins score. 34 patients had excellent or good results. Pain was retreated, patients could come back to their normal activities, before the first luxation. Only 4 patients had another luxation in spite of our treatment. The control CT showed that the humeral head retroversion was 30 or more degrees. The arthroscopy was performed, the Bankart lesion was found and the patients were qualified for treatment this lesion. Weber osteotomy could be the satisfactory method of treatment of anterior shoulder instability in patients with physiologic reduction of humeral head retroversion. PMID- 15751718 TI - [Usefulness of distraction osteogenesis in the treatment of knee arthrosis]. AB - AIMS: The purpose of this study is the assessment of efficiency of distraction osteogenesis (DO) as a method of operative treatment of knee arthrosis with co existent varus deformity. MATERIAL AND METHODS: Authors undertook an attempt of evaluation of results of tibial corticotomy in 21 patients (25 operated knee joints) treated in Orthopaedic Clinic of Wroclaw Medical University in years 1995 2001, with the use of circular, external Ilizarov's fixator. All the operated patients underwent corticotomy of proximal tibial metaphysis followed by slow axis correction and bone regenerate formation started at 7th postoperative day. In orthopaedic examination functional evaluation of knee joint with modified point scale according to Ranawat (HSS-score) there were taken into account. In rentgenometric examination the limbs axis in the frontal plane and joint space morphology according to Ahlback were evaluated. RESULTS: The results in our group of patients were very encouraging. Accordingly to Ranawat scale, in majority of patients assessed before the treatment as poor and fair moved to group assessed as fair and good and even excellent. We observed decelerated regenerate formation and remodeling probably caused by advanced age of patients in 6 cases. In 3 cases persistent lose of entire correction was observed. CONCLUSIONS: In author's opinion DO as a method of operative treatment of unicompartmental knee arthrosis could be advocated as alternative method to traditional tibial osteotomy considering its biological effect stimulating regeneration of all tissues including cartilage even in advanced unicompartmental arthrosis. PMID- 15751719 TI - [AGC DA prosthesis in severe deformity and revision knee arthroplasty]. AB - The AGC DA replacement has been used in our department for latest 3 years. 28 arthroplasties were performed on 26 patients. There were 20 women and 6 men of age from 29 to 74 (average 58,7). Bilateral knee replacements were performed on one man and one woman. 18 patients from this group were diagnosed as RA, 7 OA, and 1 LED. Primary knee replacement were done on 15 knees and secondary on 13 using AGC DA type. 26 patients were examined after half a year follow up according to Clinical Rating System of The Knee Society: 21 patients had good (75%), 6 satisfactory (21.4%) and 1 poor (3.6%) results. The results after secondary replacements were generally worse then primary ones. That was mainly due to limited ROM and walking ability. There were no early complications in wound healing although 11 patients had prolonged steroids therapy and 7 of them had also methotrexate combined with steroids given as a basic pharmacological treatment of systemic disease. There were two other complications. One patient with loosening of tibial part because of primary tibial component malpositioning. Reoperation and correcting position with longer stem was performed after 8 months. Second patient with extensor mechanism insufficiency caused luxation of prosthesis in flexion. Extensive exercises of extensor muscles solved the problem. PMID- 15751720 TI - [Thromboembolic and inflammatory complications of knee joint alloplasty with cemented endoprostheses]. AB - In the presented paper the authors evaluate the long-term results of treatment for inflammatory and thromboembolic complications in knee joint alloplasty with cemented endoprostheses. The clinical material spanning the years 1998-2003 are 15 patients with analysed complications, it determine 9% of all operated. They have evaluated achieved results using the HSS knee rating scale in their own modification. As far as the authors are concerned, venographic examination is a prerequisite for appropriate care of the patients following knee joint alloplasty who might have deep venous thrombosis. It is necessary to provide similar treatment when developing infections are suspected in the perioperative period and in cases of late infections. Consequently, antibiotic therapy and a possible quick bacteriological checkup for healing a postoperative wound is a prerequisite for successful treatment. Achieved results induce statement, that thromboembolic and inflammatory complications caused at most operated chronic vein insufficiency, deficit of range of motion at operated joint and gait insufficiency. Thromboembolic complications was always related with inflammation of knee joint alloplasty in our own material. PMID- 15751721 TI - [Remodeling after in situ pinning for slipped capital femoral epiphysis]. AB - We reviewed 61 hips in 50 children (33 boys and 17 girls) with slipped capital femoral epiphysis (SCFE), all treated by in situ pinning. Average age of our patients was 11.8 years (range, from 8 to 15 years). The average clinical and radiographic follow-up was 7.4 years. By studying the changes in physeal-shalt angle on radiographs in Lauenstein (frog) position, we found evidence of physeal remodeling in three (8%) of 38 mild and seven (30%) of 23 moderate slips. Remodeling of the proximal femoral metaphysis occurred in 92% of hips with a head shaft angle less than 30 degrees and in 52% of hips between 30 degrees and 60 degrees. A process of local resorption and apposition of bone seems to be essential to final remodeling. The probability of remodeling was significantly less the greater the degree of slip, but was significantly increased among children 11 years old or younger and if the triradiate cartilage was open at the time of presentation. The range of internal rotation was significantly greater in those hips that remodeled. Pinning in situ of a slipped capital femoral epiphysis represents a simple, quick and effective method of treatment, allowing remodeling in mild and moderate slips. PMID- 15751722 TI - [Dislocation after total hip arthroplasty]. AB - From 1997 to 2002 in the Department of Traumatology there had been performed 488 total hip replacements: 334 in men and 454 in women at the age between 20 and 84 (average 63.6). The procedures were performed by 12 surgeons and postero-lateral approach was used. Dislocation was found in 22 patients (4.51%): 14 women and 8 men. Majority of dislocation occurred during first 8 weeks after an operation and 68% of them happened during first 6 weeks. The surgical treatment was necessary in 10 cases (45% of all dislocation): 3 patients had only a surgical reposition and 7 patients had a revision arthroplasty. There was found that most cases of dislocation took place at the age above 81 years. Dislocation occurred 5 times more frequently in patients operated on for a femoral neck fractures then for coxarthrosis. The prosthesis TT1 most often dislocated possibly due to its shape (low Head Neck Ratio). Based on our own experience we find dislocation is a multifactorial complication and precise description of causes is usually very difficult. PMID- 15751723 TI - [Metacarpophalangeal joint dislocation of the thumb in children]. AB - This publication presents 7 children with traumatic dislocation of the thumb, treated in University Children's Hospital in Krakow between 1993-2002. The mean age during injury was 9 years, mean observation time 2 years. One child was operated, the others had non-operative treatment. All gained good result. The literature about this dislocation has been discussed in this paper. PMID- 15751724 TI - [Percutaneous tenotomy of achilles tendon in the treatment of congenital clubfeet -a preliminary report]. AB - The authors present early results of conservative treatment in congenital clubfeet connected with the percutaneous tendon Achilles tenotomy in 10 children (16 feet). 9 feet were of type III and 7 of type IV according to the Dimeglio scale. Minimum follow up was 6 months, with an average follow up period of 10 months. All cut Achilles tendons regenerated completely. On follow up 9 feet were completely corrected, 5 presented with mild forefoot adduction and 2 feet required surgery because of recurrent equinovarus deformity. The authors claim that the percutaneous tendon Achilles tenotomy is a safe and valuable procedure which allows to avoid early surgery in 88% of clubfeet. PMID- 15751725 TI - [Skipping of the ulnar nerve and the medial head of the triceps--a case report]. AB - The authors describe rare phenomenon of the snapping of the medial head of the triceps muscle and recurrent dislocation of the ulnar nerve. The indications for operative treatment are established. The authors describe its own case making a conclusion that operative treatment is a good option in some cases. PMID- 15751726 TI - [Biomechanics of scoliosis]. AB - Scoliosis is defined as a three-dimensional deformity of the spine. The most pronounced component of scoliosis is in the frontal plane, comprising the lateral bending of the spine. Rotation of vertebra takes place in the transverse plane. In most cases of idiopathic scoliosis a decrease of thoracic kyphosis in the sagittal plane occurs. A more rare event is the appearance of a junctional kyphosis between the primary and secondary curve. The instrumentation introduced by Harrington dealt mainly with balancing the bending forces in the frontal plane (distraction of the concavity of the curve), along with fusion of the instrumented area. The multisegmental CD instrumentation allowed for the diminution of the lateral curve in the frontal plane, while at the same time "forcing" an increase of thoracic kyphosis in single curves, and restoration of physiological sagittal curves (thoracic kyphosis lumbar lordosis) in double curve scoliosis. The CD method achieved this good by a 90 degrees rotation of the rod towards the concavity of the curve, "changing" the lateral curve into kyphotic curve. In the AO USS (Universal Spine System) correction is achieved by pulling the hooks towards the rod. The procedure ends with the linking of two rods with transverse connectors forming this way a stable framework. The degree of correction achieved with this method is based on the biomechanic inter relation between the spine and the instrumentation system (application of distraction forces, compensatory forces and translocation of the instrumented segment). Post op decompensation of the spine is usually the result of incorrect hook fixation, inadequate application of forces (distraction and compression) and use of a standard hook pattern for thoracic curves (type III) in other types of scoliosis. PMID- 15751727 TI - [Some aspects of spine biomechanics and their clinical implications in idiopathic scoliosis]. AB - The spine with it's physiological curves is designed to transfer loads to the pelvis and the lower limbs. The complex interplay of forces in the spine can be described by breaking down these forces into their basic components. These forces have a direction and can therefore treated as vectors in three axis, related to the anatomic planes: frontal, sagittal and transverse the static spine forces responsible for a correct posture create tension. Tension is defined as the relation of the vector force to the surface area to which the force is applied. Tension, depending an the direction of the force applied can be normal or tangent. Gravitational force is transferred by the vertebral bodies. Posterior elements of the spine (the lamina, the processes and ligaments) are stabilizing elements. The articular processes bear loads only when lateral bending of the spine. The complex nature of the scoliotic deformity usually leads toa decompensated spine even prior to surgery. Although considerable correction of the curve can be achieved using modern instrumentation systems, the spine can be restored. Lach of decompensation or decompensation of the spine is a major problem among many patients treated surgically with systems based on hooks and rods. Such decompensation in the frontal plane can be a result of correction beyond the compensatory possibilities of the lumbar spine, inadequate placing of hooks and incorrectly applied distraction forces. Overlooking the proximal junctional kyphosis (between the two proximal thoracic curves or overlooking the distal junctional kyphosis (between the lumbar and thoracic curve) can also lead to decompensation of the spine in the sagittal plane. PMID- 15751728 TI - [Report from the 4th International Symposium on Pediatric Orthopedics and from the training sessions in San Diego (1-27 December 2003)]. PMID- 15751729 TI - President's message. PMID- 15751730 TI - One nurse's story. PMID- 15751731 TI - The clinical nurse leader (CNL) role. PMID- 15751732 TI - Secondhand smoke fact sheet. PMID- 15751733 TI - Professional liability insurance: straight answers to your questions. PMID- 15751734 TI - Integrating healing modalities into nursing practice. PMID- 15751735 TI - Pathogenic bacteria: how to get them back into the line of fire? PMID- 15751737 TI - HIV-1 assembly and budding as targets for drug discovery. AB - Research conducted during the past several years has led to an increased understanding of the roles played by the viral protein Gag and specific cellular factors in HIV assembly/budding. The identification of compounds that interfere with this process validates this late step in virus replication as a target for HIV therapeutic discovery. In this review, current understanding of HIV-1 assembly/budding is described and several developmental stage drugs that target this process are reviewed. PMID- 15751736 TI - Protecting APOBEC3G: a potential new target for HIV drug discovery. AB - Human immune cells possess a built-in mechanism that could potentially block the replication of retroviruses such as HIV-1. This protective mechanism centers on apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G), a DNA-editing enzyme produced by host cells infected by certain retroviruses that is then incorporated into virions. Due to its cytidine deaminase activity, APOBEC3G is able to mutate the minus-strand DNA formed during reverse transcription. These events ultimately halt completion of the HIV life cycle. Unfortunately, HIV-1 encodes a protein termed virion infectivity factor (Vif) that specifically suppresses the activity of APOBEC3G. Vif achieves this effect by depleting the intracellular stores of APOBEC3G, thus making this antiviral enzyme unavailable for incorporation into budding virions. APOBEC3G depletion involves the recruitment of a specific E3 ligase complex by Vif leading to the polyubiquitylation and proteasome-mediated degradation of this enzyme. The potent activity of APOBEC3G has led to considerable interest in the identification of small molecules that interrupt the Vif-induced degradative process. PMID- 15751738 TI - Current approaches to developing a preventative HIV vaccine. AB - During the past 20 years, the pendulum of opinion in the HIV-1 vaccine field has swung between two extremes, initially favoring the induction of antibodies only, and subsequently favoring the induction of cell-mediated immune responses only. At present, the consensus seems to be that induction of both humoral and cellular immunity by an HIV-1 vaccine will be required to achieve maximum protection. One obstacle to the development of an effective HIV-1 vaccine has been the difficulty in inducing broadly reactive, potent antibodies with protective functions. Defining epitopes and designing immunogens that will induce these antibodies is one of the main challenges that currently confronts the HIV-1 vaccine field. PMID- 15751739 TI - Recent advances in antileishmanial drug development. AB - Leishmania are protozoan parasites responsible for a spectrum of diseases collectively known as leishmaniasis. The disease is a significant health problem in many regions of the world and emerges as a serious co-infection in HIV positive individuals. Current treatment of the disease is based on a limited number of chemotherapeutic agents which are rapidly becoming ineffective, and are characterized by high toxicity and cost. This review focuses on recent advances in antileishmanial drug development and improvements to current treatment options. Novel approaches currently used to identify leishmanicidal compounds as diverse as antimicrobial peptides and natural plant extracts are described in this review. PMID- 15751740 TI - Hepatotoxicity of antifungal agents. AB - Antifungal agents have been implicated in numerous cases of hepatotoxicity throughout the past few decades. Hepatotoxic reactions to antifungal agents range from slight, asymptomatic abnormalities in liver function tests to potentially fatal fulminant hepatic failure. Clinically significant hepatic injury resulting from antifungal therapy most commonly manifests as acute hepatocellular, cholestatic or mixed hepatocellular-cholestatic reactions. In general, reactions usually resolve on cessation of therapy, but some antifungal agents may induce chronic liver damage. This review will summarize the hepatotoxicity profiles of the major classes of antifungal agents and will provide recommendations for drug monitoring in order to minimize the risk of hepatotoxicity. PMID- 15751741 TI - The use of quorum-sensing blockers as therapeutic agents for the control of biofilm-associated infections. AB - The development of novel antimicrobial compounds is required to treat the growing number of infections where antibiotic resistance is a serious threat, especially in situations where biofilms are involved. Antibiotic resistance is the result of two factors: first, through the development of specific antibiotic resistance, due to either mutation or the acquisition of antibiotic resistance genes; and second, by the innate tolerance of bacterial biofilms. Bacterial control, through the inhibition of bacterial cell-cell communication systems which are involved in the regulation of virulence factor production, host colonization, and biofilm formation, is discussed in this review. Specifically, this review presents current studies on the development of quorum-sensing inhibitors for the control of bacterial infections. PMID- 15751742 TI - New strategies for the treatment of infections associated with prosthetic joints. AB - Treatment of infections associated with orthopedic devices usually requires appropriate surgical intervention combined with a prolonged course of antimicrobial therapy. The choice of optimal management depends on duration and pathogenesis of infection, stability of the implant, antimicrobial susceptibility of the pathogen and condition of the soft tissue. The role of rifampin in combination with quinolones, minocycline, co-trimoxazole or fusidic acid in the treatment of staphylococcal infections is outlined in this review. Increasing antimicrobial resistance requires the use of alternative agents, such as quinupristin-dalfopristin, linezolid and daptomycin, but results of clinical trials are limited. Finally, agents with a potential for treatment or prophylaxis of device-associated infections, such as novel rifamycin derivatives, and compounds acting on bacterial quorum-sensing mechanisms are presented in this review. PMID- 15751743 TI - Bacterial efflux pump inhibition. AB - Drug efflux is recognized as a clinically relevant mechanism of antimicrobial drug resistance in selected species of bacteria. Membrane-based polypeptides belong to distinct protein superfamilies, the members of which are related by structural characteristics, mechanism of action and energy source for the transport process. These membrane-based polypeptides mediate drug efflux, and some of these proteins, the multidrug efflux proteins, are capable of extruding numerous structurally dissimilar antimicrobial agents and biocides. Inhibition of these pumps can decrease intrinsic resistance, reverse acquired resistance and reduce the emergence of mutants with higher-level target-based mutational resistance. In vitro and limited in vivo investigations have shown that combining efflux pump inhibitors with antimicrobial agents that are pump substrates can result in the recovery of clinically relevant activity of those compounds. The identification of broad-spectrum efflux pump inhibitors may reduce the need, and thus cost, of discovery and development of new antimicrobial agents that are not pump substrates. PMID- 15751744 TI - RotaTeq (sanofi pasteur/Wistar Institute/Children's Hospital of Philadelphia). AB - Sanofi pasteur MSD (a joint venture between Merck & Co and sanofi pasteur (formerly Aventis Pasteur)), in collaboration with the Wistar Institute and the Children's Hospital of Philadelphia, is developing RotaTeq, an oral liquid pentavalent vaccine for the potential prevention of rotavirus infection-related disorders, such as infant diarrhea and malabsorption. Phase III trials had commenced by March 2001. PMID- 15751745 TI - VRX-496(VIRxSYS). AB - VIRxSYS is developing VRX-496, a lentiviral HIV-based vector encoding anti-HIV antisense envelope sequences, as a potential gene therapy for HIV infection. In July 2003, VIRxSYS undertook the initial dosing of an HIV-positive patient in a phase I/IIa trial. PMID- 15751746 TI - Telavancin (Theravance). AB - Theravance is developing telavancin, an injectable peptidoglycan inhibitor antibiotic for potential use in the treatment of Gram-positive bacterial infection. Phase III trials in complicated skin and skin structure infections commenced in September 2004. PMID- 15751747 TI - Gut flora in health and disease: potential role of probiotics. AB - In a young evolving science, there are always more questions than answers. That is also the situation in the emerging field of Probiotics, and this was made very clear at the International Probiotics Workshop in Amsterdam. In the report of this workshop, we present a selection of the most urgent questions in the field of probiotics. In addition, we propose a few strategies for the future of probiotics research. During the workshop, 120 experts--from disciplines including Human Nutrition, Gastroenterology, Nutritional Therapy, Cell Biology, Microbiology and Immunology--discussed new views on microbe-host interactions and the role of probiotics in prevention and alleviation of gastro-intestinal, atopic and auto-immune diseases. There is a general consensus among the experts that administering defined strains can help in preventing and curing gut flora related diseases: the first clinical trials show a promising role for probiotics. But the system is very complex, and most underlying mechanisms are still unclear. Rapid progress in this field will depend largely on the collaboration between fundamental researchers from different disciplines and medical specialists. Besides, more clinical studies are required to convince authorities and the public of the value of microbial therapies. PMID- 15751748 TI - Intestinal flora of animal models of human diseases as an environmental factor. AB - Genetically-engineered animals are known to be useful in clarifying the functions of many genes and as animal models for human diseases. However, it has been widely reported that pathophysiology is not expressed in these animals when they become germfree or SPF animals, i.e., the pathophysiology is not the result of genes alone and a combination of gene function and intestinal flora as an environmental factor are necessary. It is important to determine the roles of each of these two factors by pathophysiological analysis. Gnotobiotic mice were produced by establishment of specified bacterial species in germfree animals to form the intestinal flora of SPF animals and they were placed in barrier facilities. Measures have been taken against infections by bacteria such as Pseudomonas aeruginosa and Enterobacter cloacae. In addition, gnotobiotic mice with a highly normal physiology are required. Analysis of the effects of each bacterial species and combinations of bacteria on in vivo functions, i.e., the cross-talk between the host and intestinal flora, is essential in the creation of better laboratory animals. Monitoring of the intestinal flora, a key factor in the colonies produced, is a topic for future research. PMID- 15751749 TI - Bacterial diversity of Escherichia coli in the gut: a reason to re-evaluate probiotic formulations? AB - Humans and animals are increasingly being subjected to various probiotic formulations with the claim of providing a number of health benefits to the consumer. These formulations usually incorporate bacterial consortia comprising of mostly lactic acid bacteria (LAB). Recent studies have shown that strains found in different regions of the gut are genetically different from each other and may therefore have different abilities to interact with bacteria that they come into contact with. Even LAB show differences in their ability to interact, and further, inhibit growth of pathogenic bacteria in vitro due to individual strain differences. If these results are repeatedly shown to be true in future assessments, an evaluation of bacterial consortia used in probiotic formulations may now be necessary. This may have an impact in the way future probiotic formulations are prepared. PMID- 15751751 TI - Erythromycin-induced resistance to clindamycin in Staphylococcus aureus. AB - PURPOSE: To describe the incidence of erythromycin-induced resistance to clindamycin in a sample of Staphylococcus aureus isolates. METHODS: 100 erythromycin-resistant and clindamycin-sensitive S. aureus were collected as a convenience sample from February to August 2003. Inducible clindamycin resistance was identified using the D-zone disc method. RESULTS: Of the 100 Staphylococcus aureus isolates, 64 were methicillin sensitive (MSSA) and 36 were methicillin resistant (MRSA). Of the 64 MSSA isolates, 22 (34%) had inducible resistance. Of the 36 MRSA isolates, 4 (11%) had inducible resistance. Overall, 26% of these clindamycin sensitive S. aureus isolates, exhibited inducible resistance to clindamycin. CONCLUSIONS: In this sample, MSSA isolates were almost three times more likely to have inducible MLS resistance compared to MRSA isolates. Inducible resistance may compromise the efficacy of clindamycin. The frequency of inducible resistance in this series of "clindamycin sensitive" S. aureus isolates is 26%. It is likely that the true percentage of clindamycin resistance is being underestimated since testing for inducible resistance is not routinely performed. PMID- 15751752 TI - Insulinoma, a rare neuroendocrine tumor: a case report. AB - We report a case of Insulinoma, a rare neuroendocrine tumor with an incidence of approximately four per 5 million. This case demonstrates the characteristic clinical, biochemical and histological features of an insulinoma, a rare benign neuroendocrine tumor where early recognition is important to ensure proper surgical treatment and prevent serious adverse consequences. PMID- 15751753 TI - Concomitant cerebral and coronary arterial gas emboli in a sport diver: a case report. AB - This report presents a case of concomitant cerebral and coronary gas emboli seen in a sport scuba diver after suffering from pulmonary barotrauma. Except for massive fatal gas embolism, no case of concomitant cerebral and coronary arterial gas emboli has been reported. The 45 year old male diver rapidly surfaced from a depth of 32 feet of sea water and experienced transient loss of consciousness, chest pain, and hemiparesis. EKG and cardiac enzymes suggested myocardial ischemia. He received three recompression treatments and recovered completely. PMID- 15751754 TI - Self-reporting of internal medicine house staff work hours. AB - INTRODUCTION: The 80-hour workweek became a reality for residency programs nationwide on July 1, 2003. In this review of administrative data, we examine the self-reporting of work hours by a cohort of Internal Medicine residents. METHODS: Data was collected from 27 residents in training at Tripler Army Medical Center over a 4 month period from September 1 to December 31 2002. House staff reported their hours on a daily basis by responding to an email message, as well as on a monthly basis utilizing the Army's UCAPERs (Uniform Chart of Account Personnel System) mandatory monthly workload tracking system. Data from the two separate reporting systems was compared for accuracy, completeness and internal consistency. RESULTS: Compliance with daily reporting was variable (67-97% with overall compliance rate of 86%) but lower when compared with the mandatory military monthly reporting system (95-100%). There were large differences in reporting of average weekly work hours among individual residents when monthly reporting was compared to daily reporting of data with higher averages with monthly data reporting. Weekly totals averaged nearly 12 hours higher when reported monthly compared to reporting on a daily basis (p < 0.0001). A total of 18 residents reported that they worked more than 80 hours per week during one month using monthly data, while only 7 reported that they averaged more than 80 hours with the daily reporting data. When average weekly hours reported on a daily basis were compared with the total number of inpatient days worked over the four month period using a simple regression model, there was a significant relationship with average hours increasing with increasing number of inpatient days worked (adjusted R square = 0. 19, p = 0.01). CONCLUSIONS: Little internal consistency was found in the comparison of daily versus monthly work hour reporting, indicating that self-reporting may not provide accurate data. Complying with the 80-hour workweek is crucial for residency programs to maintain accreditation, and thus programs will need a way to accurately capture consistent resident work hour data. Further studies are indicated to determine the most accurate way of assessing house staff work hours. PMID- 15751756 TI - Emphysematous pyelonephritis: a case report. PMID- 15751757 TI - The flood at the John A. Burns School of Medicine. PMID- 15751758 TI - Cancer in sea turtles. PMID- 15751759 TI - Chronic hepatitis C and genotyping: the clinical significance of determining HCV genotypes. AB - Chronic hepatitis C, attributed to infection with hepatitis C virus (HCV), is a global health problem. The overall prevalence of viral hepatitis worldwide is estimated to be 3-5% with over 175 million people infected with HCV. Clinically, HCV can establish a persistent, chronic infection contributing to progressive liver disease, including cirrhosis and hepatocellular carcinoma (HCC), requiring intensive treatment regimens, possible liver transplantation and long-term care. Due to the chronic nature of HCV infection and the tremendous burden on healthcare resources, clinicians and laboratorians have looked for key epidemiological, pathological and viral characteristics that may provide insight into disease progression, severity and response to therapy to permit the administration of effective therapeutic regimens as well as long-term management of infected individuals. Determination of viral genotype has been identified as one parameter that could provide direction in the clinical management of patients with chronic HCV infections. The following review provides background on determination of HCV genotypes and the relevance of viral genome characterization in the current clinical setting. PMID- 15751760 TI - Challenges for the clinical development of new nucleoside reverse transcriptase inhibitors for HIV infection. AB - There is a need for new antiretroviral drugs with activity against HIV isolates resistant to currently available agents and improved short and long-term tolerability profiles. Clinical trial designs for nucleotide and nucleoside reverse transcriptase inhibitors (NRTIs) are restricted by the characteristics of these agents (for example, their cross-resistance, resistance threshold and interaction profiles), the ethical need to ensure that patients are not maintained on suboptimal regimens, and regulatory requirements (for example, with regards to trial designs and patient populations). For example, consideration of cross-resistance profiles must influence the way in which an NRTI in development is sequenced to minimize any impact on future treatment options. The resistance threshold is determined by the number of mutations required to diminish sensitivity to a given drug. Pharmacokinetic or pharmacodynamic interactions restrict how NRTIs may be combined during clinical development. Doses may be selected on the basis of results from short-term monotherapy studies in treatment naive patients, but such studies cannot establish the long-term efficacy or tolerability of new agents used in combination regimens. Confirmatory studies in treatment-naive populations do not meet the medical and regulatory needs for clinical data in treatment-experienced populations, while studies in treatment experienced populations are subject to numerous clinical and logistical difficulties. Intensification, switch and hybrid study designs all offer suitable approaches to the evaluation of NRTIs with novel resistance profiles. Switch studies are particularly useful for agents with resistance profiles that suggest a specific sequencing approach in treatment and for those with the potential, based on pharmacokinetic data, for interactions with other agents. The successful development of new NRTIs will depend upon a thorough appreciation of these many and complex issues, not only among those involved in the design of clinical studies, but also those contributing to their review and conduct. PMID- 15751761 TI - Nucleoside analogue-sparing strategy for the treatment of chronic HIV infection: potential interest and clinical experience. AB - Nucleoside analogue-sparing antiretroviral combinations may be interesting as first-line therapies as they spare a complete class of drugs that will remain fully active for later use and prevent the risk of mitochondrial toxicity related to exposure to nucleoside reverse transcriptase inhibitors (NRTIs). This strategy is also used in patients failing NRTIs with cross-resistance to compounds in this class. Different combinations of antiretroviral drugs are theoretically available. Non-nucleoside reverse transcriptase inhibitors (NNRTIs) associated with protease inhibitor (PI) and boosted double-PI combinations have been studied through small, non-comparative clinical studies and preliminary results suggest that they are efficient and often well-tolerated. However, NNRTIs and PIs are extensively metabolized in the liver through cytochrome P450, leading to pharmacokinetic interactions; a good knowledge of the interactions between NNRTIs and PIs, or between PIs, is helpful in assisting physicians in clinical practice in choosing drugs and doses. Access to a therapeutic drug monitoring service to confirm that appropriate drug exposures are achieved is useful when using such regimens. Some negative kinetic interactions may lead to complicated combinations with a high pill burden that reduces their applicability. Gastrointestinal toxicity often remains a limiting factor in the use of boosted double-PI combinations. Non-comparative studies have allowed selection of NRTI-sparing options that now need to be compared with the current standard of care in comparative clinical trials before being considered as valuable options. Other NRTI-sparing therapeutic strategies are emerging: PI monotherapy with lopinavir/ritonavir has been evaluated in a small group of naive patients and appears promising. Drugs belonging to new classes currently under investigation, such as entry inhibitors, might be included early in the antiretroviral treatment of patients as soon as compounds with a convenient route of administration are available, increasing the number of therapeutic combinations without NRTIs. PMID- 15751763 TI - Depression and clinical progression in HIV-infected drug users treated with highly active antiretroviral therapy. AB - OBJECTIVE: To disentangle the impact of adherence from that of injecting drug status and depressive syndrome on HIV clinical progression in a cohort of highly active antiretroviral therapy (HAART)-treated HIV patients infected through drug use. DESIGN: MANIF 2000 is a French cohort of HIV-infected drug users with scheduled medical visits every 6 months. Only patients enrolled in the MANIF 2000 cohort who had a CD4 cell count >200 cells/microl at HAART initiation were selected. The follow-up period included all post-HAART initiation visits. METHODS: HIV clinical progression was defined as either AIDS-related death or reaching a CD4 level <200 cells/microl. Adherence was assessed using a self administered questionnaire and a structured face-to-face interview. Depressive symptoms were evaluated by a Center for Epidemiologic Studies Depression Scale (CES-D) score at each visit. Cox proportional hazards model was used to calculate crude and adjusted relative hazards and 95% confidence intervals and thus identify independent predictors of clinical progression. RESULTS: Of the 305 HAART-treated patients in the cohort, 243 had CD4 cell count >200 cells/microl at HAART initiation. At the first visit after HAART initiation, median CD4 cell count was 466 cells/microl and 45% had undetectable viral load. Injecting drug users accounted for 17% of the study group. Over the follow-up period, 32 patients experienced HIV clinical progression. Probable depression was encountered in 46% of patients and non-adherence in 31% of the sample. After adjustment on baseline CD4 cell count, predictors of clinical progression were: having a higher level of cumulative non-adherence over the follow-up period [HR (95% CI)=1.2 (1.1-1.3) per 10% increase] and having a high score of depressive symptoms following HAART initiation [HR (95% CI)=5.3 (2.21-3.0)]. CONCLUSIONS: Although depressive syndrome is known to influence non-adherence behaviours that are amongst the major reasons for clinical progression, it is also a predictor of clinical progression in HIV-infected intravenous drug users on HAART, independently of non-adherence behaviours. HIV care providers should be more sensitive to depressive symptoms in order to detect them early and supply HIV patients with specific care. Further research is needed to determine whether treating depressive symptoms may improve adherence and thus delay disease progression and mortality. PMID- 15751762 TI - Optimal frequency of CD4 cell count and HIV RNA monitoring prior to initiation of antiretroviral therapy in HIV-infected patients. AB - CONTEXT: Guidelines regarding the frequency of CD4 cell count and HIV RNA monitoring in HIV-infected patients vary, with recommended strategies ranging from every 2 to every 6 months. OBJECTIVE: To determine optimal CD4 cell count and HIV RNA monitoring frequency in HIV-infected patients prior to antiretroviral therapy initiation. DESIGN: Cost-effectiveness (CE) analysis using an HIV simulation model incorporating CD4 cell count and HIV RNA as immunological and virological predictors of clinical outcomes. SETTING: Hypothetical clinical setting. PATIENTS: Simulated cohort based on initial clinical presentation of HIV infected patients in the US. INTERVENTION: CD4 cell count and HIV RNA monitoring at frequencies ranging from every 2 to 24 months prior to antiretroviral initiation, as well as accelerated monitoring frequencies as CD4 cell counts approach a specified treatment threshold. OUTCOME MEASURES: Life expectancy, quality-adjusted life expectancy and costs. RESULTS: For patients presenting with median CD4 cell count 546/mm3 and median HIV RNA 4.8 log10 copies/ml, incremental CE ratios ranged from US$37800/quality-adjusted life year (QALY) gained for a constant testing frequency of every 18 months compared with every 24 months, to US$303300/QALY gained for a constant testing frequency of every 2 months compared with every 4 months when starting treatment at a CD4 cell count of 350/mm3. Monitoring every 12 months until a warning CD4 cell count threshold of 450/mm3 followed by every 3 months until 350/mm3 had an incremental CE ratio of US$74700/QALY gained. When starting antiretroviral therapy at CD4 cell count 200/mm3, monitoring every 12 months until 300/mm3 followed by every 2 months until treatment initiation yielded an incremental CE ratio of US$52200/QALY gained compared with the next best strategy. Increasing monitoring frequency as CD4 cell counts approached a treatment threshold yielded greater incremental clinical benefit for less cost than strategies using a constant frequency. CONCLUSIONS: Monitoring HIV-infected patients every 12 months until 100 CD4 cells/mm3 prior to a specified treatment threshold followed by more frequent monitoring every 2 or 3 months until antiretroviral therapy initiation is both more effective and cost-effective than the current standard of care. PMID- 15751764 TI - Relationship between changes in thymic emigrants and cell-associated HIV-1 DNA in HIV-1-infected children initiating antiretroviral therapy. AB - OBJECTIVES AND METHODS: To investigate the relationship between cell-associated HIV-1 dynamics and recent thymic T-cell emigrants, HIV-1 DNA and T-cell receptor rearrangement excision circles (TREC, a marker of recent thymic emigrants) were measured in peripheral blood mononuclear cells in 181 samples from 33 HIV-1 infected children followed for 96 weeks after antiretroviral therapy (ART) initiation. RESULTS: At baseline, HIV-1 DNA was higher in children with higher TREC (P=0.02) and was not related to age, CD4 or HIV-1 RNA in multivariate analyses (P>0.3). Overall, TREC increased and HIV-1 DNA decreased significantly after ART initiation, with faster HIV-1 DNA declines in children with higher baseline TREC (P=0.009). The greatest decreases in HIV-1 DNA occurred in children with the smallest increases in TREC levels during ART (P=0.002). However, this inverse relationship between changes in HIV-1 DNA and TREC tended to vary according to the phase of HIV-1 RNA decline (P=0.13); for the same increase in TREC, HIV-1 DNA decline was much smaller during persistent or transient viraemia compared with stable HIV-1 RNA suppression. CONCLUSIONS: Overall, these findings indicate that TREC levels predict HIV-1 DNA response to ART and suggest that immune repopulation by thymic emigrants adversely affects HIV-1 DNA decline in the absence of persistent viral suppression, possibly by providing a cellular source for viral infection and replication. PMID- 15751765 TI - Body fat changes among antiretroviral-naive patients on PI- and NNRTI-based HAART in the Swiss HIV cohort study. AB - BACKGROUND: Body fat changes are common in patients with HIV. For patients on protease inhibitor (PI)-based highly active antiretroviral therapy (HAART), these changes have been associated with increasing exposure to therapy in general and to stavudine in particular. Our objective is to show whether such associations are more or less likely for patients on non-nucleoside reverse transcriptase inhibitor (NNRTI)-based HAART. METHODS: We included all antiretroviral-naive patients in the Swiss HIV Cohort Study starting HAART after April 2000 who had had body weight, CD4 cell count and plasma HIV RNA measured between 6 months before and 3 months after starting HAART, and at least one assessment of body fat changes after starting HAART. At visits scheduled every 6 months, fat loss or fat gain is reported by agreement between patient and physician. We estimate the association between reported body fat changes and both time on therapy and time on stavudine, using conditional logistical regression. RESULTS: Body fat changes were reported for 85 (9%) out of 925 patients at their first assessment; a further 165 had only one assessment. Of the remaining 675 patients, body fat changes were reported for 156 patients at a rate of 13.2 changes per 100 patient years. Body fat changes are more likely with increasing age [odds ratio (OR) 1.18 (1.00-1.38) per 10 years], with increasing BMI [OR 1.06 (1.01-1.11)] and in those with a lower baseline CD4 cell count [OR 0.91 (0.83-1.01) per 100 cells/microl]. There is only weak evidence that body fat changes are more likely with increasing time on HAART [OR 1.16 (0.93-1.46)]. After adjusting for time on HAART, fat loss is more likely with increasing stavudine use [OR 1.70 (1.34-2.15)]. There is no evidence of an association between reported fat changes and time on NNRTI therapy relative to PI therapy in those patients who used either one therapy or the other [OR 0.98 (0.56-1.63)]. CONCLUSION: Fat loss is more likely to be reported with increasing exposure to stavudine. We find no evidence of major differences between PI and NNRTI therapy in the risk of reported body fat changes. PMID- 15751766 TI - A randomized controlled trial to enhance antiretroviral therapy adherence in patients with a history of alcohol problems. AB - OBJECTIVE: To assess the effectiveness of an individualized multicomponent intervention to promote adherence to antiretroviral therapy (ART) in a cohort of HIV-infected individuals with a history of alcohol problems. DESIGN: We conducted a randomized controlled trial to compare the usual medical follow-up with an adherence intervention. SETTING: The principal enrolment site was Boston Medical Center, a private, not-for-profit, academic medical institution. SUBJECTS: HIV infected patients with a history of alcohol problems on ART. A total of 151 were enrolled and 141 (93%) were assessed at follow-up. INTERVENTION: A nurse, trained in motivational interviewing, completed the following over 3 months in four encounters: addressed alcohol problems; provided a watch with a programmable timer to facilitate pill taking; enhanced perception of treatment efficacy; and delivered individually tailored assistance to facilitate medication use. MAIN OUTCOME MEASURES: Prior 30-day adherence > or =95%, prior 3-day adherence of 100%, CD4 cell count, HIV RNA and alcohol consumption, each at both short- and long-term follow-up. RESULTS: At follow-up, no significant differences in medication adherence, CD4 cell count, HIV RNA or alcohol consumption were found (all P values >0.25). CONCLUSIONS: A multicomponent intervention to enhance adherence among HIV-infected individuals with a history of alcohol problems was not associated with changes in medication adherence, alcohol consumption or markers of HIV disease progression. The failure to change adherence in a group at high risk for poor adherence, despite utilizing an intensive individual-focused patient intervention, supports the idea of addressing medication adherence with supervised medication delivery or markedly simplified dosing regimens. PMID- 15751767 TI - HIV-1 infection leads to increased HLA-E expression resulting in impaired function of natural killer cells. AB - HIV has evolved several strategies to evade recognition by the host immune system including down-regulation of major histocompatibility complex (MHC) class I molecules. However, reduced expression of MHC class I molecules may stimulate natural killer (NK) cell lysis in cells of haematopoietic lineage. Here, we describe how HIV counteracts stimulation of NK cells by stabilizing surface expression of the non-classical MHC class I molecule, HLA-E. We demonstrate enhanced expression of HLA-E on lymphocytes from HIV-infected patients and show that in vitro infection of lymphocytes with HIV results in up-regulation of HLA-E expression and reduced susceptibility to NK cell cytotoxicity. Using HLA-E transfected K-562 cells, we identified the well-known HIV T-cell epitope p24 aa14 22a as a ligand for HLA-E that stabilizes surface expression of HLA-E, favouring inhibition of NK cell cytotoxicity. These results propose HIV-mediated up regulation of HLA-E expression as an additional evasion strategy targeting the antiviral activities of NK cells, which may contribute to the capability of the virus in establishing chronic infection. PMID- 15751768 TI - Response to HAART and GB virus type C coinfection in a cohort of antiretroviral naive HIV-infected individuals. AB - The prognostic role of GB virus type C (GBV-C) viraemia in HIV-infected subjects treated with highly active antiretroviral therapy (HAART) is still undefined. The aim of this analysis is to assess the relationship between GBV-C infection and response to antiretroviral therapy among HIV-infected subjects initiating HAART when antiretroviral-naive. A prospective, observational study of 400 HIV-infected patients with measurements of GBV-C RNA, hepatitis C virus (HCV) antibodies and HCV RNA determined from plasma stored prior to HAART initiation. Time to virological (achieving HIV RNA < or =500 copies/ml) and immunological success (a CD4+ count increase of > or =200 cells/microl), and the time to virological relapse (confirmed HIV RNA >500 copies/ml) were assessed by Kaplan-Meier methods and Cox proportional hazard regression model. Of the subjects, 117 (29.3%) were GBV-C positive and, overall, 351 (87.8%) patients achieved virological success. After controlling for a number of confounders including HCV RNA, GBV-C viraemic patients experienced a significantly lower risk of HIV rebound than those who were GBV-C negative [relative hazard (RH)=0.56, 95% CI: 0.34-0.93, P=0.03]. Conversely, the probability of achieving initial virological success or CD4+ count response after HAART did not differ between GBV-C-negative and -positive subjects. These results suggest that GBV-C coinfection may play a role in determining the rate of HIV rebound possibly by competing with HIV replication after HIV load has been successfully suppressed by HAART. PMID- 15751770 TI - Viral interaction and responses in chronic hepatitis C and B coinfected patients with interferon-alpha plus ribavirin combination therapy. AB - BACKGROUND/AIMS: We conducted a case-control study to investigate the efficacy of interferon-alpha (IFN-alpha) and ribavirin combination therapy for patients with chronic hepatitis C and B virus (HCV/HBV) coinfection and to elucidate the interaction of these two viruses. METHODS: Forty-two chronic HCV/HBV-coinfected patients (29 IFN-naive, 13 IFN-relapsed) and 84 HCV-monoinfected controls, matched for age, sex and previous history of IFN-alpha therapy, were enrolled. All patients were treated with IFN-alpha-2b 6 MU three-times weekly plus ribavirin 1000-1200 mg daily for 24 weeks. Serum HCV RNA and HBV DNA were determined every 24 weeks for 72 weeks. RESULTS: The rate of HCV sustained virological response (SVR) was comparable among IFN-naive and IFN-relapsed HCV/HBV-coinfected patients and IFN-naive and IFN-relapsed HCV-monoinfected patients (69.0%, 69.2%, 67.2% and 57.7%, respectively; intention-to-treat analysis). HCV genotype 1b, high pretreatment HCV RNA levels and liver fibrosis were significantly associated with a lower HCV SVR. Of 16 baseline HBV viraemic patients, five (31.3%) achieved HBV SVR, which correlated negatively to HCV genotype non-1b and HCV SVR. Only one (6.3%) had simultaneous seroclearance of HCV and HBV. Antibodies to HBV surface antigen seroconversion developed in five (11.9%) patients during long-term follow-up. HCV responders had significantly higher rates of HBV DNA resurgence than HCV non-responders during and after treatment. Reciprocal viral interference was noted between HCV and HBV after IFN alpha/ribavirin therapy. CONCLUSIONS: IFN-alpha/ribavirin combination therapy is effective for HCV/HBV-coinfected patients in eradicating HCV infection and might promote HBV seroclearance, and there is a mutual viral response and reciprocal viral interaction between HBV and HCV. PMID- 15751769 TI - Similar reduction of cytomegalovirus DNA load by oral valganciclovir and intravenous ganciclovir on pre-emptive therapy after renal and renal-pancreas transplantation. AB - BACKGROUND: Pre-emptive treatment of CMV infection in transplant recipients aims at prevention of clinical disease by early detection. However, current treatment requires the intravenous (iv) administration of ganciclovir for 2 weeks, which is a considerable burden for the patient. In this observational study, the efficacy of the new oral prodrug valganciclovir was compared with iv ganciclovir. METHODS: To facilitate the introduction of valganciclovir, a therapeutic guideline was developed to use this drug under controlled conditions with regard to safety in renal/renal-pancreas transplant recipients requiring CMV therapy. Subsequently, a group of 57 consecutive transplant recipients was evaluated. Onset and treatment of CMV infections were followed by frequent monitoring of CMV DNA in plasma by quantitative real-time PCR. Details of antiviral therapy were documented. RESULTS: In 15 out of 57 transplant recipients, a total of 27 anti-CMV treatment episodes were recorded: 18 with valganciclovir (900 mg twice daily) and nine with iv ganciclovir (5 mg/kg twice daily) as initial treatment. Median CMV DNA load reduction during treatment was 0.12 log10/day in the valganciclovir group and 0.09 log10/day in the ganciclovir group. There were no haematological side effects in any group and no patient developed signs of clinical CMV disease. CONCLUSION: Similar reduction of CMV DNA load was observed during pre-emptive treatment with oral valganciclovir and iv ganciclovir in transplant recipients. Oral valganciclovir would provide an attractive and safe alternative for pre emptive CMV treatment in renal/renal-pancreas transplant patients, however, confirmation in larger randomized studies would be desirable. PMID- 15751771 TI - Influence of rosiglitazone on flow-mediated dilation and other markers of cardiovascular risk in HIV-infected patients with lipoatrophy. AB - BACKGROUND: Antiretroviral therapy for HIV infection is commonly complicated by lipoatrophy, insulin resistance and dyslipidaemia. In HIV-uninfected adults with insulin resistance or type 2 diabetes, thiazolidinediones can lower blood pressure and improve both insulin sensitivity and endothelial function. This study sought to investigate the effects of rosiglitazone on endothelial function and other markers of cardiovascular risk in patients with HIV-related lipoatrophy. METHODS: HIV-infected, lipoatrophic adults receiving antiretroviral therapy were randomized to receive either rosiglitazone 4 mg or matched placebo, twice daily. Percentage flow-mediated forearm arterial dilation (FMD%) was measured at weeks 0, 12, 24 and 48, together with other markers of vascular risk (blood pressure, lipids, glycaemic parameters, adiponectin and leptin). RESULTS: Out of 64 enrolled adults, 44 (69%) attended all visits (23 rosiglitazone, 21 placebo). Relative to placebo, at week 48, rosiglitazone decreased systolic blood pressure (8 mmHg, P=0.03), insulin (3 microIU/ml, P=0.02), insulin resistance (P=0.03) and leptin (0.6 ng/ml, P=0.02), whilst adiponectin was increased (3.3 microg/lml, P<0.0001). However, rosiglitazone increased total cholesterol (49.1 mg/dl, P=0.001), low-density lipoprotein cholesterol (23.5 mg/dl, P=0.01) and triglycerides (146 mg/dl, P=0.06). Mean baseline FMD% for the entire cohort was moderately impaired (4.5%). Compared with baseline, mean on-treatment FMD% increased by 0.8% with rosiglitazone and decreased by 0.3% with placebo, (mean difference 1.1%, 95% CI -0.2 to 2.5, P=0.09). CONCLUSIONS: Rosiglitazone has minimal effect on flow-mediated dilation in HIV-infected lipoatrophic adults. However, despite worsening of the lipid profile, the overall effect of rosiglitazone on the cardiovascular risk profile in these subjects was positive. PMID- 15751773 TI - Prediction of HIV-1 protease inhibitor resistance using a protein-inhibitor flexible docking approach. AB - Emergence of drug resistance remains one of the most challenging issues in the treatment of HIV-1 infection. Here we focus on resistance to HIV-1 protease inhibitors (PIs) at a molecular level, which can be analysed genotypically or phenotypically. Genotypic assays are based on the analysis of mutations associated with reduced drug susceptibility, but are problematic because of the numerous mutations and mutational patterns that confer drug resistance. Phenotypic resistance or susceptibility can be experimentally evaluated by measuring the amount of free drug bound to HIV-1 protease molecules, but this procedure is expensive and time-consuming. To overcome these problems, we have developed a docking protocol that takes protein-inhibitor flexibility into account to predict phenotypic drug resistance. For six FDA-approved Pls and a total of 1792 HIV-1 protease sequence mutants, we used a combination of inhibitor flexible docking and molecular dynamics (MD) simulations to calculate protein inhibitor binding energies. Prediction results were expressed as fold changes of the calculated inhibitory constant (Ki), and the samples predicted to have fold increase in calculated Ki above the fixed cut-off were defined as drug resistant. Our combined docking and MD protocol achieved accuracies ranging from 72-83% in predicting resistance/susceptibility for five of the six drugs evaluated. Evaluating the method only on samples where our predictions concurred with established knowledge-based methods resulted in increased accuracies of 83-94% for the six drugs. The results suggest that a physics-based approach, which is readily applicable to any novel PI and/or mutant, can be used judiciously with knowledge-based approaches that require experimental training data to devise accurate models of HIV-1 Pl resistance prediction. PMID- 15751772 TI - Nevirapine and efavirenz pharmacokinetics and covariate analysis in the 2NN study. AB - OBJECTIVE: The aim of this 2NN pharmacokinetic substudy was to investigate the population pharmacokinetics of nevirapine and efavirenz. METHODS: Treatment naive, HIV-1-infected patients received nevirapine (once or twice daily), efavirenz or a combination with lamivudine and stavudine. Blood samples were collected on day 3 and weeks 1, 2, 4, 24 and 48. Using non-linear mixed effects modelling, pharmacokinetics of nevirapine and efavirenz and factors involved in the inter-individual variability were investigated. RESULTS: Clearance of nevirapine in the induction phase (<14 days) and at steady state (>28 days) were 2.02 I/h and 2.81 I/h, respectively. Volume of distribution and absorption rate constant were 77.0 l and 1.66 h(-1), respectively. Clearance of nevirapine was lower in females (13.8%) and in patients with hepatitis B (19.5%). Patients from South America and Western countries had higher clearance of nevirapine compared with Thai and South African patients. The clearances of efavirenz in the induction phase and at steady state were 7.95 l/h and 8.82 l/h, respectively. The volume of distribution and absorption rate constant were 4181 and 0.287 h(-1), respectively. Concomitant use of nevirapine increased clearance of efavirenz (43%). Patients from Thailand had lower clearance than the rest of the population. CONCLUSIONS: The population pharmacokinetics of nevirapine and efavirenz were assessed in the 2NN trial. For both drugs, an induction phase was distinguished from the steady-state phase. Gender, hepatitis B and geographical region were involved in the variability of the pharmacokinetics of nevirapine. Region and concomitantly used nevirapine were determinants of the pharmacokinetics of efavirenz. PMID- 15751774 TI - Response to interferon-based therapies in HIV-infected patients with chronic hepatitis C due to genotype 4. AB - BACKGROUND: The hepatitis C virus (HCV) genotype is the main predictor of response to interferon (IFN)-based therapies. HCV genotype 4 is spreading among European intravenous drug users, who are frequently coinfected with HIV. Information about treatment response in this subset of patients is scarce and conflicting results have been reported. METHODS: All HIV-infected patients treated for chronic hepatitis C at our institution with a minimum follow-up of 6 months after discontinuing therapy were retrospectively analysed. They had received one of three HCV treatment modalities: IFN monotherapy, IFN plus ribavirin (RBV) or pegylated interferon (PEG-IFN) plus RBV. Treatment responses were stratified according to HCV genotype. RESULTS: A total of 390 patients were analysed. Sustained virological response (SVR) to HCV therapy had been reached by 90 (23.1%): 22/119 (18.5%) with IFN monotherapy; 17/106 (16%) with IFN plus RBV; and 51/165 (30.9%) with PEG-IFN plus RBV. SVR was significantly higher among those with HCV genotypes 2 or 3 (40.4%; 61/151) than in patients with either HCV genotype 1 (11.2%; 22/197) or HCV genotype 4 (16.7%; 7/42) (P<0.0001). In contrast, there were no significant differences in the response rate comparing HCV genotypes 1 and 4 (P=0.53). CONCLUSIONS: Response to IFN-based therapies in HIV-positive patients with hepatitis C due to HCV genotype 4 is poor, similar to that obtained for HCV genotype 1 and much lower than for HCV genotypes 2 and 3. Therefore, HIV-infected patients with hepatitis C due to genotype 4 should be considered as a particular subset of difficult-to-treat patients. New treatment strategies and drugs for these patients are eagerly awaited. PMID- 15751775 TI - Early virological failure with a combination of tenofovir, didanosine and efavirenz. AB - OBJECTIVE: To describe the occurrence of a high early virological failure (VF) rate and development of resistance mutations in antiretroviral-naive patients receiving tenofovir, didanosine and efavirenz. METHODS: HIV-infected antiretroviral-naive patients with viral load > or =30 000 copies/ml were enrolled in a pilot randomized trial of tenofovir/didanosine (250 mg)/ efavirenz with (arm A) or without (arm B) lopinavir/r for the first 12 weeks. As six cases of early VF (a drop of <2 log at month 3, or a rebound of >1 log from the nadir) were detected (five in arm B and one in arm A who had previously stopped lopinavir/r) an unplanned interim analysis was performed. RESULTS: A total of 29 out of 36 enrolled patients completed at least 3 months of follow-up and were included in the interim analysis. An intent-to-treat analysis showed treatment failure in 7/15 (46.7%) patients in arm B (five VF, one lost, one switched) versus 2/14 (14.3%) in arm A (one lost, one switched) (P=0.109). The patient in arm A who interrupted lopinavir/r at day 3 and continued with tenofovir/didanosine/efavirenz later developed VF. At baseline, 6/6 VF patients had VL >100000 copies/ml and an advanced stage of disease (CD4 <200 plus CDC stage C or B3) versus 0/8 non-VF patients taking the triple drug regimen (P<0.001). At failure, G190S/E alone or associated with K103N and K101R mutations was detected in five patients, and K103N/L1001/V108l in the sixth patient. Additionally, L74V/I and K65R were detected in four and two patients, respectively. CONCLUSIONS: A high early virological failure rate and the occurrence of resistance mutations were detected in a group of antiretroviral naive patients treated with tenofovir/didanosine/efavirenz. PMID- 15751776 TI - News in hepatitis C: Report from the 39th annual meeting of the European Association for the Study of the Liver. PMID- 15751777 TI - Tubular cell apoptosis and cidofovir-induced acute renal failure. AB - Cidofovir is an antiviral drug with activity against a wide array of DNA viruses including poxvirus. The therapeutic use of cidofovir is marred by a dose-limiting side effect, nephrotoxicity, leading to proximal tubular cell injury and acute renal failure. Treatment with cidofovir requires the routine use of prophylactic measures. A correct knowledge of the cellular and molecular mechanisms of cidofovir toxicity may lead to the development of alternative prophylactic strategies. We recently cared for a patient with irreversible acute renal failure due to cidofovir. Renal biopsy showed tubular cell apoptosis. Cidofovir induced apoptosis in primary cultures of human proximal tubular cells in a temporal (peak apoptosis at 7 days) and concentration (10-40 microg/ml) pattern consistent with that of clinical toxicity. Apoptosis was identified by the presence of hypodiploid cells, by the exposure of annexin V binding sites and by morphological features and was associated with the appearance of active caspase-3 fragments. Cell death was specific as it was also present in a human proximal tubular epithelial cell line (HK-2), but not in a human kidney fibroblast cell line, and was prevented by probenecid. An inhibitor of caspase-3 (DEVD) prevented cidofovir apoptosis. The survival factors present in serum, insulin-like growth factor-1 and hepatocyte growth factor, were also protective. The present data suggest that apoptosis induction is a mechanism contributing to cidofovir nephrotoxicity. The prophylactic administration of factors with survival activity for tubular epithelium should be further explored in cidofovir renal injury. PMID- 15751778 TI - Changes in mitochondrial RNA production in cells treated with nucleoside analogues. AB - BACKGROUND: To investigate mitochondrial (mt) toxicity of antiretroviral drugs further, we developed a novel real-time PCR-based assay for the quantification of mtRNA. We analysed the effects of stavudine (d4T), didanosine (ddl) and zidovudine (AZT) on the production of mtRNAs in different human cell lines and compared the production with the amount of mtDNA present in the same cells. MATERIALS AND METHODS: HUT78, CEM and U937 cells were exposed to different nucleoside reverse transcriptase inhibitors (NRTIs) for 7 days. Thereafter, nucleic acids were isolated and Taqman-based real-time PCR was used to quantify mtDNA and three different mtRNAs (ND1, CYTB and ND6 gene products). RESULTS: Different amounts of mtRNAs exist in different cell lines. When mtRNA was measured in cells exposed to an NRTI, a marked decrease was observed in cells treated with d4T, but not with ddl or AZT. Changes in mtRNA production did not always correspond to modifications in mtDNA content: 1 microM d4T significantly changed mtRNA but not mtDNA content. CONCLUSIONS: d4T, but not ddl or AZT, significantly alters mtRNA quantity and quality. The method we have developed can reveal changes that are not observed by measuring mtDNA content only, and can be used for ex vivo studies on drug toxicity. PMID- 15751779 TI - Evolution of protease and reverse transcriptase inhibitor resistance-associated mutations in HIV-1-infected protease inhibitor-treated patients with persistent low viraemia. PMID- 15751780 TI - Guidelines for management of impacted canines. AB - Though the management of impacted canine is considered a multi-disciplinary entity requiring the expertise of a number of specialists, the general practitioner needs to coordinate care and must know about the diagnosis and management of canine impaction. The management of canine impactions can be simplified by following a logical and systematic approach. The schematic treatment plan for impacted canines and the factors influencing and determining the plan are discussed in this article. PMID- 15751781 TI - Criteria for management of calcific metamorphosis: review with a case report. AB - Calcific metamorphosis is seen commonly in the dental pulp after traumatic tooth injuries and is characterized by deposition of hard tissue within the root canal space. Opinion differs among practitioners as to whether to treat these cases upon early detection of calcific metamorphosis or to observe them until symptoms or radiographic signs of pulpal necrosis are detected. In this article, the clinical, radiographic, and histopathologic appearance of calcific metamorphosis is described; a review of the literature is presented to address these issues in an attempt to establish sound rationale for treatment. Approximately 3.8% to 24% of traumatized teeth develop varying degrees of calcific metamorphosis. Studies indicate that of these, approximately 1% to 16% will develop pulpal necrosis. Most of the literature does not support endodontic intervention unless periradicular pathoses is detected or the involved tooth becomes syptomatic. It may be advisable to manage cases demonstrating calcific metamorphosis through observation and periodic examination. A report of a case where in non-surgical endodontic intervention was successfully carried out a patient suffering from calcific metamorphosis with periapical pathoses is also presented. PMID- 15751782 TI - An audit of drug prescribing practices of dentists. AB - OBJECTIVES: Data are scarce on the prescribing habits of dental practitioners. Drug use in dentistry and stomatology is undertaken to determine the pattern of drug use for patients seeking treatment. METHODS: 1820 prescriptions of dental patients attending the dental outpatient departments at Manipal Teaching Hospital, Nepal were collected by a random once weekly survey between March '01 to February '02. The information was compiled, scored and analyzed in consultation with dentists using WHO guidelines. RESULTS: Males numbered 801 (44%) and females 1019 (56%). Most of the patients were aged between 13-25 years. The dental disorders most frequently reported in our study were diseases of pulp and periapical tissue (36.5%), gingivitis and periodontal disease (28.5%), and dental caries (16%). The average number of drugs prescribed was 2.03 (3698 / 1820) and 66% prescriptions contained antimicrobials (1 or 2). 21% drugs were prescribed in generic names and 38% drugs were fixed dose combinations of 2 or more drugs. 4/5 of the prescribed drugs were systemic agents and 1/5 were local/topical agents. The most commonly prescribed systemic agents were analgesics (43.7%) followed by antimicrobials (39%). The most conmonly prescribed local / topical agents were anti-infectives (74%). In the present study, NSAID's (89.6%) were the preferred analgesics over narcotic analgesics (10.4%). The most frequently prescribed systemic analgesic and antimicrobials were ibuprofen and amoxycillin, respectively. CONCLUSIONS: This study may help in identifying the problems involved in therapeutic decision making. Also, there is a clear need for the development of prescribing guidelines and educational initiatives to encourage the rational and appropriate use of drugs in dentistry. PMID- 15751783 TI - Comparative evaluation of the effect of palatal vault configuration on dimensional changes in complete denture during processing as well as after water immersion. AB - Polymerization shrinkage is one of the major drawbacks of the acrylic resin. The present study was conducted to evaluate the dimensional changes occurring in the dentures, constructed on the casts having different configurations of the palatal vault, during processing and the effect of water sorption on the processed dentures. Eight maxillary edentulous casts having U, F and V configurations of palatal vault were selected and complete dentures were fabricated. Dimensional changes occurring in the processed dentures were studied in different regions during processing as well as after water sorption. It was observed that dimensional changes in frontal and vertical planes were maximum in V shaped palatal vault dentures. Water sorption partly compensated for the dimensional changes that occurred during processing. PMID- 15751784 TI - Canalicular adenoma of the upper lip mimicking mucocele. AB - Salivary gland neoplasms are relatively rare. Among the salivary gland neoplasms minor salivary gland neoplasms are often deceptive to the clinician and challenging to the pathologist because of their varied morphology and diverse histopathological pattern. This article documents a case of canalicular adenoma of the upper lip mimicking mucocele and discusses the differential diagnosis of minor salivary gland neoplasms. PMID- 15751785 TI - Malignant melanoma of the oral cavity--case reports and review of literature. AB - Melanoma is a malignant tumor of melanocytes or their precursor cells. It is the third most common skin cancer and is one of the most biologically unpredictable neoplasms. Sunlight is an important etiological factor in cutaneous melanoma. Oral melanoma is relatively uncommon, exhibiting a definite predilection for palate and maxillary gingiva, and is twice as common in males as in females. We herewith report three cases of oral melanoma out of which two occurred on the palate and one in the maxillary gingiva. Their clinical presentations, histopathology and management are discussed. PMID- 15751786 TI - Mucormycosis causing extensive destruction of maxilla. AB - Mucormycosis is a rare, often fatal opportunistic infection caused by a saprophytic fungus belonging to a class of Phycomycetes. It is characterized by its unrelenting progression towards vital organs with marked propensity towards arterial wall by direct extension producing vascular thrombosis leading to ischaemic necrosis. In normal circumstances the saprophytes are not pathogenic but represent opportunist-requiring impairments in patient's resistance to disease. If not treated the disease may prove fatal. We report a case of Mucormycosis with extensive necrosis of the maxilla in a 57-year-old patient suffering from uncontrolled diabetes with ketoacidosis. PMID- 15751787 TI - Gingival cyst of adult--a case report with unusual findings. AB - Gingival cysts of adult are rare odontogenic cysts of developmental origin. They occur as swellings on the gingiva, most commonly between 5th and 6th decade of life and usually do not exceed 1cm in size. The epithelial lining of these cysts are thin non-keratinised and are thought to arise from the rests of dental lamina. We report a case of multicystic variant of a gingival cyst in adult patient with few unusual findings regarding the age of occurrence, size of the lesion and the histopathological appearance. Gingival cyst of adult occurring in an 18-year-old male patient, measuring about 3 x 4 sq. cms on the facial attached gingiva in relation to 42, 43, and 44 was observed. The histopathological findings were that of the rare multicystic variant with keratinised lining and epithelial plaques in the lining. The lumen of the cyst showed keratin with one of the lumen showing calcifications within it. This case documents many interesting and unusual findings in gingival cyst of adults. It appears to be the first documentation of gingival cyst of adult containing calcifications within the cystic lumen. PMID- 15751788 TI - [In situ gelling polymers for ophthalmic drops]. AB - One of the possibilities of how to optimize bioavailability of drugs after topical ophthalmic administration is prolongation of the period of contact of the preparation with the eye tissue. The most usual way of achieving this gaol is an increase in the viscosity of the preparation by an addition of a suitable auxiliary substance. At present besides hydrogels and mucoadhesive substances, also polymers which produce the sol-gel phase change are employed for this purpose. This so-called in situ gelation takes place in a very short period of time after contact with the eye tissue due to a change in temperature, pH, or interaction with physiological ions. The present paper aims to survey in situ gelling polymeric substances, which are examined in relation to their use in ophthalmic administration. PMID- 15751789 TI - [The present and future of controlled transport of the drug into the organism]. AB - The problems of the systems of controlled (and thus also targeted) transport of the drug into the organism is one of the burning questions of contemporary biogalenics. Various more or less successful solutions were elaborated and applied, beginning with the retardette via the so-called intelligent hydrogels to various automatically controlled implants. Modem microfluidic intelligent biomicrosystems, composed of mutually interconnected microtanks, microjets, micropumps, or microcylinders, are very promising. A natural component of the complex biomicrosystem is a sensory subsystem for the collection of information from the bio-environment and a processor for the control of the process of drug supply to the organism. The present technologies of such Bio-Micro-Electro Mechanical Systems (BioMEMSs) make it possible to revolutionize drug transport not only by facilitating precise dosing and long-term control of the immediate amount of the supplied drug on the basis of the current condition of the patient, but also to target the drug to the site of its pharmacological effect. Though at present there are still many unsolved problems, transition from laboratory conditions to clinical practice has started, and it is only a matter of five to ten years that intelligent BioMEMSs will gradually become a routinely used dosage microform. The paper briefly surveys the present state and the next development of intelligent systems of drug supply into the body of the patient, termed Intelligent Drug Delivery Systems, Intelligent DDSs, or briefly IDDSs. PMID- 15751790 TI - [Enzymology of production of benzylisoquinoline alkaloids]. AB - This review paper summarizes the current knowledge of enzymes participating in the production of benzylisoquinoline alkaloids. This group of alkaloids comprises, e.g., morphine, codeine, thebaine, and sanginarine, which have an irreplaceable position in pharmaceutical practice. For the time being, chemists have not managed to prepare them synthetically with sufficient efficacy, and therefore the study of the enzymology of their formation remains a topical problem. The paper pays particular attention to the knowledge of individual enzymes on the molecular, or gene level. This very knowledge is essential for possible introduction of molecular-genetic approaches to the cultivation of plants producing therapeutically interesting benzylisoquinoline alkaloids. PMID- 15751791 TI - [Adsorption of substances on active charcoal. Part 2: basic esters of phenylcarbamic acid with beta-adrenolytic effect]. AB - The paper is concerned with the study of adsorption on active charcoal in a set of selected substances from the group of aryloxyaminopropanol derivatives with carbamate substitution on the benzene ring with beta-adrenolytic effect (Group A) and a set of substances, derivatives of [(arylcarbonyl)oxylaminopropanol with the identical, but assumed ultra-short effect (Group B), where the effect was produced by replacing the phenolether group with a metabolically unstable ester functional group. The course of adsorption in buffer solution with pH 7 in dependence on time and concentration is examined. Adsorptivity of substances is evaluated according to Freundlich and Langmuir models. Affinity of substances of Group A to adsorption material decreases with increasing hydrophilicity. PMID- 15751792 TI - [Prescription and structure of formulation of medicinal preparations in a pharmacy. II]. AB - The paper is concerned with the analysis of all medicinal preparations prepared extemporaneously in a pharmacy within 12 months (July 2002 through June 2003). The set under study comprised 5 260 preparations. Preparations were prescribed primarily by dermatologists, their share being 31.3 %. The expiratory period of the largest number of preparations (61.8 %) was one month. Of the total number of medicinal preparations, 76.1% were prepared on medical prescriptions, 3.5 % on orders, and 20.4 % were sold over the counter. The average annual share of preparations in the total turnover was 2.3 %. PMID- 15751793 TI - [Analysis of drugs used in out-patient practice of veterinary medicine]. AB - Both veterinary and human mass-produced medicinal preparations can by employed in the therapy of animals. Veterinary surgeons are allowed to dispense drugs right in their surgeries. Data were obtained by means of questionnaires from 11 respondents in the year 2002. Variability and consumption of mass-produced medicaments were evaluated in altogether 27 pharmacotherapeutic groups (17 groups were represented both in the assortment of veterinary and human brand products, 4 groups included only human brand products, and 6 groups were exclusively veterinary brand products). The greatest variability of human brand products prescribed in veterinary practice was found in antibiotics, vitamins, and analgesics including antiphlogistics. Of veterinary brand products, the greatest variability was found in antibiotics, anti-ectoparasitic agents, and minerals including calcium preparations. In the assortment of human brand products, the largest consumption is that of corticosteroids, antibiotics, and analgesics including antiphlogistics. In the assortment of veterinary brand products, the largest consumption was found in anti-ectoparasitic, antibiotic, and antiparasitic-anthelmintic drugs. The number of extemporaneous preparations prescribed by the veterinary surgeons ranged from 0 to 30 per month, the number of brand products dispensed in pharmacies was 0-40 of human brand products and 0 5 of veterinary brand products per month. The number of original packages of brand products stored in surgeries of veterinary surgeons ranged within 20-150 of human brand products and 40-400 of veterinary brand products. Nearly one half of the respondents reported that they did not need any cooperation with the pharmacist. PMID- 15751794 TI - [Power law equation and liberation kinetics]. AB - For the modelling of the kinetics of diffusion and relaxation in the polymer matrix, the power law equation in a two-parameter or three-parameter form is employed. The introductory part of the paper summarizes the employed varieties of the power law equation including the equation making it possible to distinguish diffusion from relaxation. The liberation kinetics of pilocarpinium chloride from lyophilized lamellae with graded amounts of hydroxypropylmethylcellulose is expressed by power law equations and their parameters are compared. The conclusion discusses the usability of the power law equation in the modelling of liberation of active ingredients and recommends the use of the three-parameter power law equation with a shift of the liberation out of the origin of the coordinates. PMID- 15751797 TI - Leavitt calls for dollar 60 billion in Medicaid savings. PMID- 15751795 TI - [Antilipoxygenase activity and the trace elements content of Aloe vera in relation to the therapeutical effect]. AB - Aloe vera is a rich source of many natural-health-promoting substances. The results of contemporary research on animal models indicate that the extracts have an antiinflammatory property. In this work the results of some in vitro experiments are shown: determination of the inhibitory effect of the Aloe vera extracts on the activity of partially purified lipoxygenase from the rat lung cytosol fraction, and quantitative determination of the trace elements presented in the extract (Mn, Fe, Cu, Zn) carried out by using the x-ray fluorescence analysis. The findings could explain the inhibitory effect (antilipoxygenase activity) of the Aloe vera extract in the acute inflammation process, expecially in the topical application for healing of minor burns and skin ulcers. PMID- 15751796 TI - [Effect of manganese (II), cobalt (II), and nickel (II) ions on the growth and production of coumarins in the suspension culture of Angelica archangelica L]. AB - The plant cell reacts to an increased concentration of metals in the environment by various mechanisms. They include an increase in the formation of heat-shock proteins, metallothioneins, phytochelatins, amino acids (cysteine, histidine), organic acids (citric, malic), or secondary metabolites. The latter mechanism is being investigated for its possible use in explant cultures for the stimulation of secondary metabolism, which is the source of substances of pharmaceutical importance. The study tested manganese (II) (0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, and 50 mM in the medium), cobalt (II), and nickel (II) ions (0, 0.1, 0.5, 1, 5, 10, 50, 100, 200, and 500 microM in the medium) as potential elicitors of coumarin production. At the same time, toxicity of these metals for the culture was examined by evaluating their effect on growth (characterized by fresh and dry weight of biomass at the end of a two-week cultivation). Cultures were cultivated in the dark and in the light. It has been found that the growth of cultures is not influenced by manganese in concentrations ranging from 0 to 2 mM, then it slightly decreases, at a concentration of 50 mM it is lower by 20 % when cultivated in the dark and by 30 % when cultivated in the light in comparison with the control. Cobalt in concentrations of 0 to 50 microM does not significantly influence the growth of the culture, higher concentrations decrease the biomass yields, more markedly when cultivated in the light (at 500 microM Co by 60 %, in the dark only by 30 % in comparison with the controls). Nickel in concentrations of 0.1 to 200 microM does not influence growth, and in a concentration of 500 microM decreases it by approximately 30 % in comparison with the control both in the light and dark. Production of coumarins was not stimulated by any metal in comparison with the control cultures, only the removal of manganese from the medium in the culture cultivated in the dark increased production by about 15 % versus the control. PMID- 15751798 TI - Will the federal government offer states P4P for Medicaid? PMID- 15751799 TI - Broad assault coming on health costs ... or else. PMID- 15751801 TI - [Cost of foods fortified with plant sterols refunded by a health insurance company]. AB - The Dutch health insurance company VGZ recently decided to refund part of the cost of using spread, yogurt and milk fortified with plant sterols. Plant sterols lower plasma cholesterol concentrations but they do so less effectively than statins. For health insurance companies wishing to improve the health of their clients, it would be more logical to reimburse costs of folic acid supplements for the prevention of neural tube defects, and vitamin-D-enriched products to prevent fractures in elderly people. PMID- 15751800 TI - [The aortocaval fistula, a rare complication of an aneurysm of the abdominal aorta]. AB - Four patients, 3 men aged 73, 60 and 81 years with hemodynamic instability and 1 man aged 80 with abdominal symptoms and breathlessness appeared to have an arteriovenous fistula secondary to spontaneous rupture of an atherosclerotic aneurysm: between the aorta and the V. cava inferior or between the A. and the left V. iliaca communis. One patient died, one patient had postoperative decompensatio cordis, one suffered a deep vein thrombosis and the 4th recovered without symptoms. The presence of an aortocaval fistula has to be considered in patients with a symptomatic abdominal aneurysm with a harsh bruit heard over the abdomen, signs of high venous pressure and peripheral hypoperfusion. When no rupture of the aneurysm is found at laparotomy in symptomatic patients, the presence of a fistula is rare, but has to be considered. Furthermore, a fistula can be the underlying cause of therapy-resistant heart failure or acute renal dysfunction. Pre-operative identification can lead to decrease of morbidity and mortality of the phenomenon. PMID- 15751802 TI - [Two surgeons from the Surgeon Association in Amsterdam, the Netherlands, 1699]. PMID- 15751803 TI - [The practice guideline 'TIA' (first revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - A transient ischaemic attack (TIA) can be seen as an attack of angina of the brain. In the first weeks following a TIA there is a sizeable risk of subsequent stroke. After this period, this increased risk tapers off and an elevated risk of cardiovascular disorders remains, with the emphasis on its cardiac component. The management of a patient with a TIA in general practice focuses on these two risk periods. The long-term cardiovascular risk can easily be reduced by prescribing aspirin, an established procedure in general practice. The immediate threat of obstruction of the carotid artery may, as was only recently firmly established, be averted by a timely referral for additional diagnostics and, when needed, carotid desobstruction. This part of TIA management in general practice, however, is hampered by the current waiting periods for these procedures. PMID- 15751804 TI - [The practice guideline 'TIA' (first revision) from the Dutch College of General Practitioners; a response from the perspective of neurology]. AB - The revised TIA guideline is a significant improvement: the diagnosis 'TIA' no longer depends on the symptoms having disappeared within 24 h, the criteria for referral to a neurologist have been widened, and the antithrombotic treatments for the various manifestations of cardiovascular disease have been harmonised. PMID- 15751805 TI - [Surgical treatment of early-stage vulva carcinoma and the complications of the operation]. AB - The treatment of patients with early-stage squamous-cell carcinoma of the vulva (with a depth of invasion > 1 mm), i.e. stage T1 with a tumour diameter < or = 2 cm or T2 with a diameter > 2 cm without suspect groin nodes on palpation, has become less radical; in this way, the complications can be reduced without compromising the generally favourable prognosis. Wide local excision with tumour free margins of 2 cm appears to be a safe option for the local treatment. Uni- or bilateral inguinofemoral lymphadenectomy with separate incisions is currently part of the standard treatment. The complications associated with this standard surgical treatment remain significant: there are frequent disorders of wound healing, wound infections, lymphoceles, lymphoedema and effects on psychosexual behaviour. The minimal invasive sentinel lymph-node procedure is a promising technique in patients with early-stage squamous-cell carcinoma of the vulva, but the safety of the procedure must still be proven. PMID- 15751806 TI - [Summary of the practice guideline 'TIA' (first revision) from the Dutch College of General Practitioners]. AB - The new practice guideline of the Dutch College of General Practitioners on the management of patients with a TIA resembles the first version, but there are some important changes: The concept TIA has been narrowed to a neurological deficit that has resolved spontaneously by the time the patient consults the doctor. The indications for referral of patients with a TIA in the area supplied by the carotid artery for investigation of a possible carotid stenosis and carotid surgery have been broadened. Auscultation of the internal carotid artery is no longer necessary. The daily amount of acetylsalicylic acid to be taken by a patient with a TIA has been increased from 30 mg to 80 mg. The prescription of dipyridamol following a TIA remains controversial. PMID- 15751807 TI - [Diagnostic image (227). A 7-year-old boy with oedema]. AB - Protein-losing gastroenteropathy with oedema, Menetrier's disease, was diagnosed in a 7-year-old boy. PMID- 15751808 TI - [Reduction in the consumption of benzodiazepines due to a letter to chronic users from their own general practitioner]. AB - OBJECTIVE: To investigate the effectiveness of a minimal intervention in reducing the volume of prescription of benzodiazepines at the regional level. DESIGN: Prospective. METHOD: Patients on compulsory health insurance who had received 180 or more daily doses of benzodiazepines in the course of one year received a letter from their general practitioner (GP) with information about the risks of chronic use, the advice to reduce or stop use, and an invitation to make an appointment to discuss the problem. This intervention took place in 19 GP practices in East Groningen. A reminder was sent six months later to the patients who had not responded in 9 randomly selected GP-practices. Thirty-seven practices in East Groningen and 91 practices in Northwest Groningen served as controls. Outcome measures were: (a) the percentage of patients who stopped, and (b) the change in average benzodiazepine consumption. RESULTS: During the period 6-12 months after the first letter in the intervention group (n = 1343), 11.3% of the patients (95% CI: 9.6-13.1) received no prescription whatsoever for benzodiazepines compared to 5.4% (4.6-6.3) and 4.9% (4.2-5.5) in East Groningen (n = 2932) and in Northwest Groningen (n = 4562), respectively. The average decrease in prescription volume was 13% (-9.9 to -15.1) in the intervention group compared to 3% (-0.1 to -4.1) and 3% (-1.5 to -4) in the control groups. The reminder sent half a year later had no additional effect. CONCLUSION: The informative letter from the GP to chronic users of benzodiazepines with the advice to stop or reduce this use was effective. PMID- 15751809 TI - [Percutaneous closure of a perimembraneous ventricular septal defect: the first 4 patients in The Netherlands]. AB - In four female patients, aged 30, 10, 8 and 2 years and 9 months (the latter with Down's syndrome), all with a perimembranous ventricular septal defect (VSD), percutaneous closure of the VSD was carried out using an Amplatzer endoprosthesis. These were the first 4 patients to undergo this procedure in The Netherlands. In 3 of the patients the procedure was without complications and complete occlusion of the defects was achieved. The last patient developed a left anterior hemiblock after implantation of the device and one week later an intermittent second degree atrioventricular block was detected, for which a pacemaker was implanted. A few days later the atrioventricular conduction time returned to normal. There was complete occlusion of the defect. Percutaneous closure of a perimembranous VSD using an Amplatzer prosthesis is a promising technique with good short-term results. In a selected group of patients this new technique can replace surgical treatment. PMID- 15751810 TI - [Acute dyspnoea following transfusion of plasma-containing blood products]. AB - A 35-year-old male patient who was given chemotherapy because of chronic myeloid leukaemia became dyspnoeic after transfusion of thrombocytes; initially, no explanation could be found for this dyspnoea. He went home before all diagnostic procedures were evaluated. Chest X-ray revealed bilateral pulmonary oedema, which could be due to transfusion-related acute lung injury (TRALI), especially since there were no indications for a cardiac aetiology. The patient was sent to the nearest hospital where he was treated with diuretics and observed for 24 hours. There were no complications. The pathogenesis of TRALI has been attributed to an interaction between anti-granulocyte antibodies and granulocytes. In addition, bioactive compounds produced during the storage of blood products have been implicated. It is important to recognize TRALI as the cause of dyspnoea when cardiac or pulmonary causes are excluded. The overall prognosis is good when treatment is started in time. The management of TRALI is supportive, with mechanical ventilation when necessary. After excluding donors with proven anti granulocyte antibodies from further donation, there is no increased risk for recurrent episodes after future transfusion of plasma-containing blood products. PMID- 15751811 TI - [Chronic heart failure and depression: the limitations of specialization-specific multidisciplinary guidelines]. PMID- 15751812 TI - [Medicinal cannabis for diseases of the nervous system: no convincing evidence of effectiveness]. PMID- 15751813 TI - Methotrexate in systemic lupus erythematosus. AB - The most appropriate treatment of moderate, nonorgan threatening disease activity in systemic lupus erythematosus (SLE) remains poorly defined, although methotrexate (MTX) is commonly used. The results of 20 uncontrolled case series and a single retrospective cohort study tend to support its use to treat active skin and joint disease. Unfortunately, three prospective randomized trials have reported conflicting results. Two reported improvement in overall disease activity and decreased corticosteroid requirement with MTX. The third trial showed no benefit from MTX for disease activity, but did report a reduction in corticosteroid requirements. Difficulties in conducting trials in moderately active SLE are discussed. PMID- 15751814 TI - Bone mineral density in postmenopausal Chinese patients with systemic lupus erythematosus. AB - The objective was to study the bone mineral density (BMD) and its clinical determinants in a cohort of postmenopausal patients with systemic lupus erythematosus (SLE). All postmenopausal SLE patients receiving long term glucocorticoids were identified from our medical clinics. Lumbar and femoral BMDs were measured by dual X-ray absorptiometry. Clinical determinants of BMD were studied by simple and multiple linear regression. Variables evaluated were: age, body mass index, parity, duration of menopause, smoking and alcohol drinking, duration of SLE and steroid treatment, cumulative prednisone dose, clinical and serological profile, disease activity, damage index and the use of medications. In total, 34 patients were studied. The mean age was 52.9+/-4.9 years and the median duration of SLE was 75.5 months. The mean duration of menopause was 5.2+/ 3.9 years and the daily maintenance dose of prednisone was 4.0+/-2.5 mg/day. At the lumbar spine, 33% of the patients were osteopenic and 48% were osteoporotic. Two patients had thoracic and lumbar vertebral compression fractures. At the nondominant femoral neck, 74% of patients were osteopenic but only 3% was osteoporotic. In a multivariate model, the current or past use of hydroxychloroquine (HCQ) was associated with a higher spinal BMD. The presence of anti-Sm and the absence of anti-Ro were associated with a higher femoral BMD. It was concluded that osteoporosis, especially at the spine, is a common and serious problem in postmenopausal Chinese SLE patients receiving long term glucocorticoid therapy. Active intervention should be considered. The protective role of HCQ has to be confirmed with further studies. PMID- 15751815 TI - Impact of external quality assessment on antinuclear antibody detection performance. AB - Our objective was to evaluate performance of the clinical laboratories for the detection of antinuclear antibodies (ANA) by using indirect immunofluorescence method (IIF), in France. A national external quality assessment (EQA) on ANA detection was organized by the French health products safety agency once a year since 1998. Between 606 to 687 laboratories together with six university reference laboratories experienced in performing tests in autoimmunity participated in the six-year consecutive survey. Each laboratory had to answer to methodological procedures and give coded responses. Variability in IIF methodological procedure was observed. Use of inappropriate microscope magnifications for reading slides or nonconventional cutoff dilution of serum were pointed out to concerned laboratories. Concerning ANA measurement, the rate of good responses ranged from 92.7% to 99.5% of the laboratories when the samples contained ANA. A wide dispersion of ANA titers obtained on a same sample was repeatly observed every year. Misinterpretation of particular fluorescence pattern was noticed. On ANA negative sample, the rate of good responses was 94.3%. In conclusion, ANA detection in routine practice is far from being standardized. However, EQA may have an impact on ANA detection performance when it is conducted on several consecutive year surveys, by providing advice for participating laboratories to limit inter laboratory variations related to methodological procedures. PMID- 15751816 TI - Efficacy and safety of nadroparin in the treatment of pregnant women with antiphospholipid syndrome: a prospective cohort study. AB - The optimal therapeutic management of patients with antiphospholipid syndrome (APS) during pregnancy is debatable. In the present prospective cohort study the use of a low molecular weight heparin (LMWH) (nadroparin), administered alone twice daily in 30 pregnant women who were diagnosed with APS on the basis of the current classification criteria, is evaluated. Dosage was adjusted according to anti-Xa levels in the patients as the pregnancies progressed. Three women, in whom an important gradual fall in platelet count in the first trimester did not respond to increased nadroparin doses, were shifted to a second-line treatment protocol. Fetal loss occurred in two of the 27 remaining women (7.40%), while 25 (92.59%) delivered 25 live infants, between the 32nd and 40th weeks of gestation. No fetal problems were registered during pregnancies, while maternal complications occurred in two of the 25 patients (8%). Moreover, there were no thrombotic events in any of the women during the study. Patient compliance was good and only minor side-effects were reported. The results of this study indicate that nadroparin alone is useful and safe in the management of pregnant patients with APS. However, in consideration of the good pregnancy outcome obtained in patients with only pregnancy morbidity when heparin and aspirin were used together, other studies comparing nadroparin twice daily with once daily plus Aspirin would be useful to ascertain which is more effective in these patients. PMID- 15751818 TI - Cardiac magnetic resonance imaging abnormalities in systemic lupus erythematosus: a preliminary report. AB - The purpose of this prospective, pilot study was to determine whether differences in myocardial T2 relaxivity can be identified among active systemic lupus erythematosus (SLE) patients with clinically suspected SLE myocarditis, other active SLE patients, inactive SLE patients and age and gender matched controls. Eleven consecutive female patients (six with active SLE and five with inactive SLE), and five age, gender and race matched healthy controls underwent imaging with echocardiography and cardiac magnetic resonance imaging (MRI). Echocardiographic measurements included left ventricular end diastolic (LVEDV) and end systolic volumes (LVESV), and mass (LVM) (all indexed to body mass); ejection fraction and cardiac output. The cardiac MRI measurement was the T2 relaxation time (an index of soft tissue signal, with higher levels suggestive of increased tissue water content). Patients with active SLE had significantly higher LVEDV and LVM than inactive SLE patients and healthy controls, and significantly larger LVESV than healthy controls. Myocardial T2 relaxation times were significantly higher in patients with active SLE compared to those with inactive SLE and to healthy controls, and remained higher even after excluding the two active SLE patients who had clinical myocarditis. The four active SLE patients who underwent repeat cardiac imaging studies after clinical improvement showed normalization of these myocardial abnormalities. The conclusion was that active SLE patients demonstrate abnormalities in myocardial structure manifested by high myocardial T2 relaxation times that normalized after clinical improvement in disease activity. These findings suggest that T2 relaxation values are a sensitive indicator of myocardial disease in patients with SLE and that myocardial T2 relaxation abnormality frequently occur in patients with active SLE, even in the absence of myocardial involvement by clinical criteria. PMID- 15751817 TI - Circulating levels of tissue factor and proinflammatory cytokines in patients with primary antiphospholipid syndrome or leprosy related antiphospholipid antibodies. AB - The antiphospholipid syndrome (APS) is characterized by the presence of antiphospholipid antibodies (aPL) in patients with thromboembolic complications. In APS, most aPL are autoantibodies to beta2-glycoprotein I and prothrombin, which play a major role in the APS pathogenesis. Nevertheless, antibodies with the same antigen specificity are also found in aPL patients with leprosy, in whom thromboembolic complications are uncommon. The in vivo upregulation of the tissue factor (TF) pathway and the imbalance of cytokines have been proposed as potential mechanisms of thrombosis in the APS. We measured the circulating levels of TF, interleukin 6 (IL-6), IL-6 receptor (sIL-6R), tumor necrosis factor (TNF alpha) and interferon gamma (IFN-gamma) in 83 patients with autoimmune aPL (42 with and 41 without clinical features of definite primary APS), 48 leprosy patients (33 with aPL) and 48 normal controls. There was a trend (P = 0.06) to higher median sTF in patients with autoimmune aPL (139 pg/mL) compared with leprosy patients (103.5 pg/mL) and controls (123 pg/mL). In addition, the frequency of raised sTF levels (> 187 pg/mL) was significantly higher in the group with autoimmune aPL [22.9% (APS 21.4%, non-APS 24.4%)] but not in leprosy (10.4%) compared with controls (4.2%). Elevated levels of IL-6 and TNF-alpha and a trend to lower IFN-gamma were found in patients with definite APS. Leprosy patients with aPL, however, had increased TNF-alpha and IFN-gamma but normal IL-6 levels. Levels of sIL-6R did not differ between controls and either patients with autoimmune aPL or leprosy. The different cytokine profiles as well as differences in circulating levels of TF might contribute to the high thrombotic risk found in patients with autoimmune aPL but not in leprosy related aPL patients. PMID- 15751820 TI - Cardio-pulmonary involvement in juvenile systemic lupus erythematosus. AB - Cardio-pulmonary manifestations of systemic lupus erythematosus (SLE) are well recognized in adults. We report the occurrence of clinically significant cardio pulmonary disease in a cohort of predominantly Caucasian children with SLE. All children with SLE attending the Royal Liverpool Children's NHS Trust between 1995 and 2003 were reviewed. Of 29 children with SLE, 27 (93%) were Caucasian. Nine (31%) had cardio-respiratory complications: cardiac only (n = 1); respiratory only (n = 4); both cardiac and respiratory manifestations (n = 4). Median (range) duration of follow-up of affected children: four years (six months to 11 years). Six out of eight (75%) presented with respiratory complications before SLE was diagnosed. Three children had pericardial effusions, one requiring pericardiocentesis for tamponade. One had cardiac conduction defects and another significant pulmonary hypertension. Respiratory complications comprised: interstitial lung disease (n = 4), with two showing evidence of pulmonary fibrosis; pleural effusions (n = 2), pulmonary haemorrhage (n = 1) and lupus pneumonitis (n = 1). Disease course was complicated by CMV infection in one child. Lung biopsy was performed in five cases. Seven were treated with cyclophosphamide with significant improvement in symptoms/lung function. Of this predominantly Caucasian paediatric cohort with SLE, 31% had significant cardio pulmonary involvement. All children with SLE should have regular monitoring of their cardio-respiratory status. PMID- 15751819 TI - Maternal and fetal outcome of lupus pregnancy: a prospective study of 29 pregnancies. AB - The aim of this study was to analyse pregestational and pregnancy risk factors for adverse fetal and maternal outcome in lupus pregnancy. Twenty women with systemic lupus erythematosus (SLE) (29 pregnancies) were prospectively evaluated. Mean patient age was 29.5+/-4.7 years, and mean disease duration, 6.3+/-6.5 years. Twenty-two pregnancies (75.9%) ended in live births; preterm delivery occurred in 17.4%, intrauterine growth restriction in 50%, preeclampsia in 3.7%, and gestational hypertension in 8%. Six pregnancies (20.7%) ended in spontaneous abortions. Adverse live-birth outcome was significantly associated with low pregestational serum albumin level, elevated gestational anti-dsDNA antibody, and diabetes mellitus. Spontaneous abortion was directly associated with low levels of pregestational serum albumin, positive anticardiolipin IgA, anti-beta2 glycoprotein I IgM, and anti-La antibodies, and inversely associated with number of patients' children. Postgestational lupus flare-up was noted in six pregnancies. Risk factors included high pregestational SLE Disease Activity Index (SLEDAI), lower serum albumin, elevated serum antibody to dsDNA, proteinuria, and use of prednisone and hydroxychloroquine. We conclude that despite high rate of obstetrical complications and postpartum lupus flare-up, pregnancy poses low risk for the majority of women with SLE. PMID- 15751821 TI - Autologous peripheral blood stem cell transplantation with in vivo T-cell depletion for life threatening refractory systemic lupus erythematosus. AB - We report the first Australian application of autologous haemopoietic stem cell transplantation in a 39-year old woman with severe systemic lupus erythematosus (SLE) and multiple life threatening complications, refractory to conventional therapy including intravenous cyclophosphamide. Our transplant technique, although not unique, differs from most published reports, in which an unmanipulated peripheral stem cell graft was used with in vivo lymphocyte depletion using rabbit antithymocyte globulin (ATG). Successful stem cell mobilization was achieved using granulocytecolony stimulating factor mobilization with methylprednisolone cover, after an initial attempt at mobilization was curtailed by respiratory arrest from upper airway obstruction due to cricoarytenoiditis, requiring tracheostomy. Conditioning regimen for the transplantation was cyclophosphamide 50 mg/kg on days -5 to -2 and rabbit ATG 2.2 mg/kg on days -3 and -2. An unmanipulated autograft was infused, with in vivo T cell depletion achieved through a further dose of ATG given on day +2 postinfusion. The autologous transplant was well tolerated without fever or other serious complication. At 12 months follow-up post-transplantation, there is an objective evidence of near-complete response with SLE disease activity index scores falling from 40 pretransplant to 2. We conclude that HSCT with unmanipulated peripheral stem cell graft and in vivo lymphocyte depletion with ATG is safe and effective therapy for cyclophosphamide refractory SLE. PMID- 15751822 TI - Hypoparathyroidism in systemic lupus erythematosus. AB - We present the case of a man diagnosed with lupus and hypoparathyroidism. This combined diagnosis has very rarely been reported, which is surprising given the evidence that idiopathic hypoparathyroidism can be due to autoimmunity, the multiplicity of organ systems that lupus can affect and the evidence that other endocrinopathies such as hypothyroidism do occur at increased rates in patients with lupus. Hypoparathyroidism does not always cause overt clinical symptoms but the resultant hypocalcemia be a significant health risk. In addition to presenting the case, we review the relevant literature and offer a discussion. PMID- 15751823 TI - Giant cell myocarditis: a rare cardiovascular manifestation in a patient with systemic lupus erythematosus. AB - Giant cell myocarditis (GCM) is a rare form of myocarditis with a median survival of less than one year. It has been reported to occur in patients with various underlying autoimmune diseases; however, no cases of GCM have been described in patients with clear evidence of underlying systemic lupus erythematosus (SLE). The presentation of GCM may mimic that of lupus myocarditis, including an initial response to immunosuppression. Despite initial clinical similarities, lupus myocarditis and GCM are histologically distinct entities with dramatic differences in prognosis. We report herein a patient with a longstanding history of SLE, who presented acutely with myocarditis, responded well to initial immunosuppression and then subsequently died of progressive heart failure that was found to be due to GCM. Endomyocardial biopsy can help define diagnosis and prognosis of lupus patients presenting with myocarditis, and early referral for cardiac transplantation should be considered in patients diagnosed with GCM. PMID- 15751824 TI - Systemic lupus erythematosus in Tunisia. PMID- 15751825 TI - Nonlupus nephritis in an adolescent with systemic lupus erythematosus. PMID- 15751826 TI - A case of Churg-Strauss syndrome with visual loss following central retinal artery occlusion. PMID- 15751827 TI - A cross-cultural investigation into the relationships between personality traits and work values. AB - Over 500 working individuals from 2 European countries (Great Britain and Greece) participated in a 2-study investigation into the relationships between the Big Five personality factors and individuals' work values. In Study 1 (N = 314), British employees completed the 60-item NEO-Five-Factor Inventory (NEO-FFI; P. T. Costa & R. R. McCrae, 1989) and the Work Values Questionnaire (WVQ; Mantech, 1983), which comprises 37 items measuring intrinsic (e.g., autonomy) and extrinsic (e.g., pay) work values. In Study 2 (N = 216), Greek employees completed a Greek language measure of the Big Five and a translated version of the WVQ. The authors observed a similar factor structure for the WVQ items in both studies. Personality traits, age, and gender explained between 5% and 13% of the variance in the WVQ subscales. As a result of the 2 studies, the authors concluded that there are robust associations between certain personality traits and work values, although they do not clearly follow the intrinsic vs. extrinsic distinction. PMID- 15751828 TI - Deadline rush: a time management phenomenon and its mathematical description. AB - A typical time management phenomenon is the rush before a deadline. Behavioral decision making research can be used to predict how behavior changes before a deadline. People are likely not to work on a project with a deadline in the far future because they generally discount future outcomes. Only when the deadline is close are people likely to work. On the basis of recent intertemporal choice experiments, the authors argue that a hyperbolic function should provide a more accurate description of the deadline rush than an exponential function predicted by an economic model of discounted utility. To show this, the fit of the hyperbolic and the exponential function were compared with data sets that describe when students study for exams. As predicted, the hyperbolic function fit the data significantly better than the exponential function. The implication for time management decisions is that they are most likely to be inconsistent over time (i.e., people make a plan how to use their time but do not follow it). PMID- 15751830 TI - Effect of the destructive disagreement belief on satisfaction with one's closest friend. AB - The purpose of this study was to test the hypothesis that, when imagining a serious disagreement with one's closest friend, verbalizing the irrational belief that disagreement is destructive would lead to greater relationship dissatisfaction than would verbalizing the rational belief that disagreement is not destructive. The author used two counter-demand-control conditions to test a second hypothesis that this effect would not be the result of demand characteristics. Undergraduates (119 women and 43 men) were randomly assigned to 1 of 5 conditions in a pretest-posttest design. In comparison with a control condition, posttest relationship dissatisfaction was significantly higher in the irrational condition and was lower, but not significantly so, in the rational condition. These results were not attributable to demand characteristics and suggested that this irrational belief increased relationship dissatisfaction. PMID- 15751829 TI - Relationships between critical thinking and attitudes toward women's roles in society. AB - This exploratory study is the first examination of the relationships between critical thinking--as measured by subscales on the Watson-Glaser Critical Thinking Appraisal Form S (WGCTA Form S; G. B. Watson & E. M. Glaser, 1994)-and attitudes toward women's roles in society--as measured by subscales on the Attitudes Toward Women Scale (AWS: J. T. Spence & R. L. Helmreich, 1972). The authors hypothesized that greater critical thinking skills would be associated with more liberal attitudes toward women's roles in society. Results from a sample of 90 nursing and management undergraduates supported the hypothesis. There were moderate correlations between scores from the WGCTA Form S Inference (.24 to .37) and Deduction (.19 to .26) subtests and scores from the AWS Freedom and Independence; Marital Relationships and Obligation; and Vocational, Educational, and Intellectual Roles subscales. These findings suggest that further research with larger samples and cross-cultural samples is warranted. PMID- 15751831 TI - The effects of prior knowledge on study-time allocation and free recall: investigating the discrepancy reduction model. AB - In this study, the authors examined the influence of prior knowledge activation on information processing by means of a prior knowledge activation procedure adopted from the read-generate paradigm. On the basis of cue-target pairs, participants in the experimental groups generated two different sets of items before studying a relevant list. Subsequently, participants were informed that they had to study the items in the list and that they should try to remember as many items as possible. The authors assessed the processing time allocated to the items in the list and free recall of those items. The results revealed that the experimental groups spent less time on items that had already been activated. In addition, the experimental groups outperformed the control group in overall free recall and in free recall of the activated items. Between-group comparisons did not demonstrate significant effects with respect to the processing time and free recall of nonactivated items. The authors interpreted these results in terms of the discrepancy reduction model of regulating the amount of processing time allocated to different parts of the list. PMID- 15751832 TI - Philosophy, self-knowledge, and personality in Iranian teachers and students of philosophy. AB - Like psychology, philosophy apparently operates from a commitment to the belief that self-knowledge should be a goal of disciplinary and personal development. Iranian teachers and students of philosophy responded to a Philosophical Orientations Scale created for this study that assessed the possible content of a high school philosophy course, along with instruments measuring self-knowledge, need for cognition, the five-factor model, anxiety, depression, and perceived stress. As the authors hypothesized, self-knowledge predicted higher levels of a philosophical orientation, even after controlling for the variance explained by need for cognition and openness to experience. Philosophical orientations and self-knowledge were also correlated with psychological adjustment, and teachers scored higher than students on these two sets of constructs. These data supported the hypothesis that personal and disciplinary interests in an adaptive self knowledge converge in philosophy. PMID- 15751833 TI - [Maternal recognition of pregnancy--comparative biology]. AB - Despite of the rapid expansion experienced by the reproductive biotechnologies in last few years, their success still limited by the high rates of early embryonic death. Since continuos fetomaternal dialogue is a prerequisite of pregnancy in all mammals, during the past decade more emphasis have been devoted to understanding the cell biological processes that regulate embryo maternal communication. This review aims to highlight the unique features of pregnancy in the different mammalian species and outlines the uterine immunological response triggered by the oocyte upon fertilization as well as the mechanisims developed by the embryo in the different species to silence or byoass the maternal immune system, induce corpus luteum maintainance and maternal recognition of pregnancy. PMID- 15751834 TI - [Epthelial derivates of the transversal groove of the stomodeum: parotid gland and juxtaoral organ]. AB - A study was carried out in humans on the initial developmental stages of structures derived from the epithelial transversal groove of the primitive mouth. In O'Rahilly's stage 16, an epithelial condensation can be observed, the anlage of Chievitz's organ, while the anlage of the parotid gland was observed in O'Rahilly's stage 18. These structures are defined by their relationships with the mesenchyme and with their topographical location. PMID- 15751835 TI - [Considerations regarding Spa treatments]. AB - Spa treatments constitute complex therapeutic processes, spanning multiple millennia. These treatments have had and retain multiple indications in the cure of multiple chronic affections, functional deficiencies, senile troubles, etc. Additionally it is omportant to consider the influence of climatic circumstances as well as social and psychic factors. The spa treatment in all its coplexity can play the role of therapeutic agent in various pathologic processes, but also be an important factor to consider in healthcare education. However, many professionals consider these procedures as an obsolete therapy of little justification today. PMID- 15751836 TI - [The moral behaviour in human an non-human primates]. AB - The well-known ethologist and Nobel Price Konrad Lorenz said that, according to the evolution, the human mental capacities, and therefore the morality, ought to have had forerunner in the animals which have preceded us: he call them <>, i.e. forerunners of our reason. Recently, the social life of non human primates has been intensely studied with the conclusions that there are rules which can be considered as rudiments of a moral behaviour. The so called <>, for instance, can be an advantage for others, but at the expense of those who practise it and this can be interpreted as a moral behaviour. PMID- 15751837 TI - [The orbitofrotal cortex I: anatomy and memory procesing]. AB - The orbitofrontal cortex has extensive tight connections with the medial temporal tobe and medial thalamic structures, which are responsible for memory processing and consolidation. What is more, this cortex is constantly activated in the memory encoding processes. This makes the orbitofrontal cortex a critical region for memory formation. This cortex is also connected with the motor and hetero- and uni- modal association sensory cortices, the limbic cortices, and subcortical structures responsible for functions related with these systems. All these facts convert the orbitofrontal cortex into a nodal region within the neural networks responsible of selecting, assembling and analyzing, based on our memory, present and pass experiences, so that we can organize and decide the most appropriate behaviour in a given situation. PMID- 15751838 TI - [New contributions to development of the cartilage of the second branchial arch and its clinical incidence]. AB - In the human embryo, the second branchial arch has a cartilaginous structure called Reichert's cartilage named alter its discoverer who first classified the visceral arches in 1837. The concept of Reichert's cartilage is still interpreted today as a structure that gives rise to the stapes of the middle ear, to the styloid process, to the styloid ligament, the lesser horns and the superior part of the hyoid body. The normal pattern of the cartilage of the second branchial arch has been studied in human fetuses, giving rise to a new interpretation. It has been classified into two parts, one longer cranial one joined to the otic capsule and another smaller caudal one associated with the hyoid bone. In our opinion, variability in the shape and the arrangement of the cranial portion will condition the formation of a styloid process of different length and morphology. Our studies of this cartilage explain the mechanism by which certain craniofacial variations with clinical implications can arise. PMID- 15751839 TI - [Michael Servetus: symphony in <>]. AB - On October 27th 1553, the physician and religious reformer Miguel Servet was burnt at stake. The 450th anniversary of his death took place, therefore, a few months ago. This is a fact which deserves being taken into account, not only because of the importance of the individual, but also, and mainly, because during the last years some important historiographical advances have changed the traditional approach, giving place to a new and more accurate analysis to his life and work. The topic image of Servet spread by the Spanish historiography of the 19th and most of the 20th Centuries has been that of a scientific genius, discoverer of the pulmonary circulation of blood, but also that of a crazy theologian with absurd ideas about some basic Christian dogmas, specially the Holy Trinity. This image is now considered absolutely misleading. The historiography of the complex religious movements of the 16th Century has shown that Servet, far from being a crazy theologian absolutely disconnected of the religious movements of the Century, was instead one of the leaders of an important Christian movement which G. H. Williams called <> He consecrated all his life to this goal, which also was the reason of his death, always showing an absolute and admirable intellectual and human coherence. To these goals he subdued any other objective, as well as the study and the exercise of medicine. This gives the basis to correctly understand the fact, never explained convincingly until now, why his great scientific discovery, the pulmonary circulation of blood, appeared as a minimal part in the heart of a theological book. PMID- 15751840 TI - [Mitral valve reconstruction: classic concepts for new indications]. AB - Mitral valve reconstructive surgery has been established as the treatment of choice in patients with favourable anatomy. Long-term follow-up has shown a better survival, lower incidence of thromboembolism, endocarditis, neurological events, lower need for mitral valve-related reoperation, and a better ventricular function after valve repair than replacement. Subvalvular apparatus repair with chordae tendineae replacement with politetrafluoretilene suture and, more recently, using mitral valve homograft has widened the indications for valve reconstruction in some patients. The classic concepts of mitral annuloplasty are now used in new indications, as for the ischemic mitral regurgitation or the functional mitral insufficiency associated to dilated cardiomyopathy. The understanding of the patho-physiology of these entities has allowed these new surgical alternatives for valve repair. PMID- 15751841 TI - [Prof. Dr. Andrew V. Schally]. PMID- 15751842 TI - [New approaches to treatment of various cancers based on cytotoxic analogs of LHRH, somatostatin and bombesin]. AB - The development of targeted cytotoxic analogs of hypothalamic peptides for the therapy of various cancers is reviewed and various oncological studies on experimental tumors are summarized. Novel therapeutic modalities for breast, prostate and ovarian cancer consist of the use of targeted cytotoxic analogs of LH-RH containing doxorubicin (DOX) or 2-pyrrolino-DOX. The same radicals have been incorporated finto cytotoxic analogs of somatostatin which can be also targeted to receptors for this peptide in prostatic, mammary, ovarian, renal cancers, brain tumors and their metastases, A targeted cytotoxic analog of bombesin containing 2-pyrrolino-DOX has been also synthesized and successfully tried in experimental models of prostate cancer, small cell lung carcinoma and brain tumors. The development of these new classes of peptide analogs should lead to a more effective treatment for various cancers. PMID- 15751843 TI - [Music as a symptom]. AB - The contents of this presentation are the consequence o reading the book Infectious Diseases and Music where the authors Drs. Gomis and Sanchez describe the infections suffered by more than fourty composers or interpreters. Although infections were more prevalent, intense psychological repercussions were also frequent. Reviewing the biographies of Bach, Mozart, Schubert and Beethoven I have selected some specially dramatic paragraphs of letters addressed for relatives and friends describing their intense and permanent physical and psychological disturbances which probably influenced the contents and style of their creations. Depression, anxiety and specially bipolar conditions with frequent and intense maniac phases were common but not exclusive to composers. Other artists and painters or poets also complained of similar disturbances. During their maiac states the artists perceive sounds and visual stimuli as well as their personal experiences with increased intensity and liveliness. Language is more fluid and their creativity and productivity become more powerful. PMID- 15751844 TI - [Historical compendium of physical activity and sport]. AB - The Historical Compendium of Physical Activity and Sport analyses, from a scientific perspective, past events in the array of tasks or manoeuvres comprising body movement, in a setting of human liberty and creative capacity. Sport is examined as a result of the evolution of games and in a context of these games. This book contemplates sports, whose selection criteria have included specific features of their individual or team qualities, or combat or opposition characteristics, or those related to their artistic features or to instrumentation or adaptation, or other connotations. 1st authors are 36 professors, from the universities of Barcelona, (Central), Granada, Jaen, Las Palmas de Gran Canaria, Leon, Madrid (Complutense), Malaga, Murcia, Oviedo, Pais Vasco, Salamanca, Valencia and Zaragoza. PMID- 15751845 TI - Wavelet packet correlation methods in biometrics. AB - We introduce wavelet packet correlation filter classifiers. Correlation filters are traditionally designed in the image domain by minimization of some criterion function of the image training set. Instead, we perform classification in wavelet spaces that have training set representations that provide better solutions to the optimization problem in the filter design. We propose a pruning algorithm to find these wavelet spaces by using a correlation energy cost function, and we describe a match score fusion algorithm for applying the filters trained across the packet tree. The proposed classification algorithm is suitable for any object recognition task. We present results by implementing a biometric recognition system that uses the NIST 24 fingerprint database, and show that applying correlation filters in the wavelet domain results in considerable improvement of the standard correlation filter algorithm. PMID- 15751846 TI - Improved fingerprint identification with supervised filtering enhancement. AB - An important step in the fingerprint identification system is the reliable extraction of distinct features from fingerprint images. Identification performance is directly related to the enhancement of fingerprint images during or after the enrollment phase. Among the various enhancement algorithms, artificial-intelligence-based feature-extraction techniques are attractive owing to their adaptive learning properties. We present a new supervised filtering technique that is based on a dynamic neural-network approach to develop a robust fingerprint enhancement algorithm. For pattern matching, a joint transform correlation (JTC) algorithm has been incorporated that offers high processing speed for real-time applications. Because the fringe-adjusted JTC algorithm has been found to yield a significantly better correlation output compared with alternate JTCs, we used this algorithm for the identification process. Test results are presented to verify the effectiveness of the proposed algorithm. PMID- 15751847 TI - Illumination-tolerant face verification of low-bit-rate JPEG2000 wavelet images with advanced correlation filters for handheld devices. AB - Face recognition on mobile devices, such as personal digital assistants and cell phones, is a big challenge owing to the limited computational resources available to run verifications on the devices themselves. One approach is to transmit the captured face images by use of the cell-phone connection and to run the verification on a remote station. However, owing to limitations in communication bandwidth, it may be necessary to transmit a compressed version of the image. We propose using the image compression standard JPEG2000, which is a wavelet-based compression engine used to compress the face images to low bit rates suitable for transmission over low-bandwidth communication channels. At the receiver end, the face images are reconstructed with a JPEG2000 decoder and are fed into the verification engine. We explore how advanced correlation filters, such as the minimum average correlation energy filter [Appl. Opt. 26, 3633 (1987)] and its variants, perform by using face images captured under different illumination conditions and encoded with different bit rates under the JPEG2000 wavelet encoding standard. We evaluate the performance of these filters by using illumination variations from the Carnegie Mellon University's Pose, Illumination, and Expression (PIE) face database. We also demonstrate the tolerance of these filters to noisy versions of images with illumination variations. PMID- 15751848 TI - Implementation of a high-speed face recognition system that uses an optical parallel correlator. AB - We implement a fully automatic fast face recognition system by using a 1000 frame/s optical parallel correlator designed and assembled by us. The operational speed for the 1:N (i.e., matching one image against N, where N refers to the number of images in the database) identification experiment (4000 face images) amounts to less than 1.5 s, including the preprocessing and postprocessing times. The binary real-only matched filter is devised for the sake of face recognition, and the system is optimized by the false-rejection rate (FRR) and the false acceptance rate (FAR), according to 300 samples selected by the biometrics guideline. From trial 1:N identification experiments with the optical parallel correlator, we acquired low error rates of 2.6% FRR and 1.3% FAR. Facial images of people wearing thin glasses or heavy makeup that rendered identification difficult were identified with this system. PMID- 15751849 TI - Three-dimensional sensor-based face recognition. AB - We describe a face recognition system based on two different three-dimensional (3D) sensors. We use 3D sensors to overcome the pose-variation problems that cannot be effectively solved in two-dimensional images. We acquire input data based on a structured-light system and compare it with 3D faces that are obtained from a 3D laser scanner. Owing to differences in structure between the input data and the 3D faces, we can generate the range images of the probe and stored images. For estimating the head pose of input data, we propose a novel error compensated singular-value decomposition that geometrically estimates the rotation angle. Face recognition rates obtained with principal component analysis on various range images of 35 people in different poses show promising results. PMID- 15751850 TI - Comparative study of face recognition techniques that use joint transform correlation and principal component analysis. AB - Face recognition based on principal component analysis (PCA) that uses eigenfaces is popular in face recognition markets. We present a comparison between various optoelectronic face recognition techniques and a PCA-based technique for face recognition. Computer simulations are used to study the effectiveness of the PCA based technique, especially for facial images with a high level of distortion. Results are then compared with various distortion-invariant optoelectronic face recognition algorithms such as synthetic discriminant functions (SDF), projection slice SDF, optical-correlator-based neural networks, and pose-estimation-based correlation. PMID- 15751852 TI - Extending the imaging volume for biometric iris recognition. AB - The use of the human iris as a biometric has recently attracted significant interest in the area of security applications. The need to capture an iris without active user cooperation places demands on the optical system. Unlike a traditional optical design, in which a large imaging volume is traded off for diminished imaging resolution and capacity for collecting light, Wavefront Coded imaging is a computational imaging technology capable of expanding the imaging volume while maintaining an accurate and robust iris identification capability. We apply Wavefront Coded imaging to extend the imaging volume of the iris recognition application. PMID- 15751851 TI - Eye-tracking architecture for biometrics and remote monitoring. AB - Eye tracking is one of the latest technologies that has shown potential in several areas, including biometrics; human-computer interactions for people with and without disabilities; and noninvasive monitoring, detection, and even diagnosis of physiological and neurological problems in individuals. Current noninvasive eye-tracking methods achieve a 30-Hz rate with a low accuracy in gaze estimation, which is insufficient for many applications. We propose a new noninvasive optical eye-tracking system that is capable of operating at speeds as high as 6-12 kHz. A new CCD video camera and hardware architecture are used, and a novel fast algorithm leverages specific features of the input CCD camera to yield a real-time eye-tracking system. A field-programmable gate array is used to control the CCD camera and to execute the operations. Initial results show the excellent performance of our system under severe head-motion and low-contrast conditions. PMID- 15751853 TI - New automated iris image acquisition method. AB - I propose a new iris image acquisition method based on wide- and narrow-view iris cameras. The narrow-view camera has the functionalities of automatic zooming, focusing, panning, and tilting based on the two-dimensional and three-dimensional eye positions detected from the wide- and narrow-view stereo cameras. By using the wide- and narrow-view iris cameras, I compute the user's gaze position, which is used for aligning the X-Y position of the user's eye, and I use the visible light illuminator for fake-eye detection. PMID- 15751854 TI - Secure authentication system that generates seed from biometric information. AB - As biometric recognition techniques are gradually improved, the stability of biometric authentication systems are enhanced. Although bioinformation has properties that make it resistant to fraud, biometric authentication systems are not immune to hacking. We show a secure biometric authentication system (1) to guarantee the integrity of biometric information by mixing data by use of a biometric key and (2) to raise recognition rates by use of bimodal biometrics. PMID- 15751855 TI - Optical security system for the protection of personal identification information. AB - A new optical security system for the protection of personal identification information is proposed. First, authentication of the encrypted personal information is carried out by primary recognition of a personal identification number (PIN) with the proposed multiplexed minimum average correlation energy phase-encrypted (MMACE_p) filter. The MMACE_p filter, synthesized with phase encrypted training images, can increase the discrimination capability and prevent the leak of personal identification information. After the PIN is recognized, speedy authentication of personal information can be achieved through one-to-one optical correlation by means of the optical wavelet filter. The possibility of information counterfeiting can be significantly decreased with the double identification process. Simulation results demonstrate the effectiveness of the proposed technique. PMID- 15751856 TI - Accurate first-order leaky-wave analysis of antiresonant reflecting optical waveguides. AB - We present a closed-form approximation for estimating both the field distribution and complex propagating constant of the antiresonant reflecting optical waveguide (ARROW) based on a first-order leaky-mode analysis. The formula was obtained from a novel coupled-electric-coupled-magnetic matrix method and provides six significant figures of the real part of the propagation constant beta of a SiO2/TiO2/SiO2/Si ARROW with an 8-microm core. The accuracy for the quantity of the imaginary part of beta is greater than 98.4% for the TE0 mode and 99.3% for TM0. The approximate values for field components are 96.1% accurate. In addition, a slight absorption by the substrate will result in modification of the initial improper leaky-mode behavior, which grows exponentially in the substrate, yielding a proper solution. PMID- 15751857 TI - Continued analysis of optical frequency-modulated continuous-wave interference. AB - I continue to analyze systematically the theory of optical frequency-modulated continuous-wave (FMCW) interference. Two special cases, multiple-beam optical FMCW interference and multiple-wavelength optical FMCW interference, are discussed in detail. Multiple-beam optical FMCW interference generates a signal with multiple frequencies because of mutual interference among the waves. Multiple-wavelength optical FMCW interference produces a signal whose amplitude is modulated by a synthetic wave. The applications of both types of optical FMCW interference are also discussed. PMID- 15751858 TI - Imaging of tumor angiogenesis in rat brains in vivo by photoacoustic tomography. AB - Green laser pulses at a wavelength of 532 nm from a Q-switched Nd:YAG laser were employed as irradiation sources for photoacoustic tomography (PAT). The vascular structure of the brain was imaged clearly, with optimal contrast, because blood has strong absorption near this wavelength. The photoacoustic images of rat brain tumors in this study clearly reveal the angiogenesis that is associated with tumors. Brain tumors can be identified based on the distorted vascular architecture of brain tumorigenesis and related vascular changes, such as hemorrhage. This research demonstrates that PAT can potentially provide a powerful tool for small-animal biological research. PMID- 15751859 TI - Configuration of an off-axis three-mirror system focused on compactness and brightness. AB - We propose a configuration of an off-axis three-mirror system for maximum compactness and brightness. The chief ray is arranged to cross three times inside the system, and the system has a round configuration for compactness. We introduced into the design a ray triangle formed by the reflection points of the chief rays at the mirrors. The ray triangle indicates the size and the brightness of the system. Based on the proposed configuration, a design example of a 4 degrees x 4 degrees field of view is shown. The F-number of the system is 2.2, in close agreement with the estimation from the ray triangle. PMID- 15751860 TI - Control of the backscattered intensity in random rectangular-groove surfaces with variations in the groove depth. AB - Recently a modification of the Kirchhoff approximation was presented to permit the calculation of multiple-scattered light on rough surfaces with infinite slopes. A brief description of the method, with examples of the scattering of light from rough surfaces with rectangular grooves, is presented. It is shown that, for a surface with random groove depths with a constant probability distribution of the groove depths, the backscatter intensity can increase or decrease, depending on the width of this probability distribution. PMID- 15751861 TI - Fiber-optic tunable multiwavelength variable attenuator and routing module designs that use bulk acousto-optics. AB - A compact fiber-coupled bulk acousto-optical multiwavelength variable optical attenuator module design that uses a retroreflective double-pass geometry within a single bulk acousto-optic tunable filter device is presented. The proposed attenuator module demonstrates a high 17-dB notch dynamic range at a low 100-mW drive power and uses a single bulk collinear-interaction acousto-optic tunable filter device. Experiments show a low (<1.8-dB) fiber-to-fiber insertion loss with a fast 34-micros speed within a wide 1520-1640-nm agile multinotch band. The basic broadband attenuator module design is extended to allow for efficient architectures for routing modules such as agile drop filters, analog hitless tap filters, and digital add-drop switches. PMID- 15751862 TI - High-power efficient continuous-wave TEM00 intracavity frequency-doubled diode pumped Nd:YLF laser. AB - We describe experimental results with a diode-pumped, intracavity-doubled cw Nd:YLF laser in multilongitudinal mode and TEM00 spatial transverse mode with a critical phase-matched lithium triborate crystal. Taking into account the thermal effects of Nd:YLF, energy-transfer upconversion, and the thermal fracture limit, we set up a power-scaling model to optimize and design a fundamental diode-pumped Nd:YLF laser. A highly efficient second-harmonic laser was achieved, based on the optimized cavity design. A second-harmonic-generation output power of 20.5 W at a wavelength of 527 nm was obtained at an incident pump power of 60 W, corresponding to an optical-to-optical efficiency of 34.2%. The TEM00 mode green laser operates at a measured M2 parameter of 1.2. The instability of the green laser power is less than +/- 1% RMS. PMID- 15751864 TI - Multipass configuration to achieve high-frequency conversion in Li2B4O7 crystals. AB - A multipass configuration for second-harmonic generation of Nd:YAG laser radiation is demonstrated to produce, for the first time to the authors' knowledge in twin lithium tetraborate crystals, as much as 21% conversion efficiency even though the effective nonlinear coefficient of the crystal is as low as 1/6th that of KDP. Apart from crystals that have large walk-off angles, low effective nonlinear coefficients, or both, the simple experimental setup would also be quite suitable for those crystals, especially infrared crystals, that have large effective nonlinear coefficients but low laser damage thresholds. PMID- 15751863 TI - Optical frequency-domain reflectometry with a rapid wavelength-scanning superstructure-grating distributed Bragg reflector laser. AB - Superstructure-grating distributed Bragg reflector lasers are particularly suited for optical frequency-domain reflectometry optical-coherence tomography with wide wavelength tunability and frequency agility. We report theoretical estimates of and experimental results for the data acquisition speed, the observable depth range, the resolution, and the dynamic range of an optical frequency-domain reflectometry system that uses a superstructure-grating distributed Bragg reflector laser whose wavelength can be tuned from 1533 to 1574 nm with a tuning speed of 10 micros/0.1-nm step. PMID- 15751865 TI - Multiwavelength investigation of laser-damage performance in potassium dihydrogen phosphate after laser annealing. AB - The laser-induced damage performance of potassium dihydrogen phosphate and deuterated potassium dihydrogen phosphate nonlinear optical crystals after pre exposure to lower-energy laser pulses (laser annealing, also known as laser conditioning) is investigated as a function of wavelength for both the damaging and conditioning pulses. We obtain a quantitative evaluation of the bulk damage performance of these materials by measuring the density of damage events as a function of laser parameters. This new method allows for a detailed assessment of the improvement of material performance from laser conditioning and reveals the key parameters for optimizing performance depending on the operational wavelength. PMID- 15751866 TI - Longitudinal pure spatial coherence of a light field with wide frequency and angular spectra. AB - We have observed the longitudinal pure spatial coherence of a light field in an interference experiment when the length of the temporal coherence is significantly smaller than the length of the longitudinal spatial coherence of the light field. We introduce into consideration new spatial and temporal scales of a light field: the length of the coherent (free) run and the coherent time (the time of life) of a wave train. PMID- 15751867 TI - Spectral Talbot phenomena of frequency combs induced by cross-phase modulation in optical fibers. AB - Cross-phase modulation (XPM) of a frequency comb (finite-duration optical pulse sequence) by an intense, long Gaussian pump pulse is theoretically investigated, and new effects, namely, frequency-domain self-imaging phenomena (integer and fractional Talbot effects), are reported. The conditions favorable for observing spectral self-imaging phenomena by XPM are derived and numerically confirmed. The effects of nonidealities in a practical experiment (e.g., group-delay walk-off and dispersion) are also evaluated. One can use spectral self-imaging to tune the free spectral range of a frequency comb (without affecting the shape and bandwidth of the individual passbands) simply by adjusting the pump power in a fiber XPM scheme. PMID- 15751868 TI - Combined effects of bending and elongation on polymer optical fiber losses. AB - The combined effects of bending and elongation on fiber losses as rays propagate along deformed polymer optical fibers (POFs) are investigated. The variations in power attenuation for various curvature radii and elongations are studied. The experimental results indicate that the combination of bending and elongation significantly affects the power loss of POF. From the results an equation is proposed to predict the power losses for different bent radii and elongations. The maximum difference between the proposed equation and the experimental results is less than 5%. PMID- 15751869 TI - D-type fiber biosensor based on surface-plasmon resonance technology and heterodyne interferometry. AB - A D-type fiber biosensor based on surface-plasmon resonance (SPR) technology and heterodyne interferometry is presented. The sensing device is a single-mode optical fiber in which half the core is polished away and a thin-film layer of gold is deposited. We measure the phase-difference variations instead of the light intensity as in traditional SPR techniques. Therefore the accuracy and resolution of our method are very high. Its sensitivity can reach 2 x 10(-6) refractive-index units. The sensor has some merits, e.g., tunable high sensitivity, small size, lower cost, smaller sample volume, and suitability for in vivo testing. This novel method of a D-type fiber biosensor based on SPR technology and heterodyne interferometry is valuable for chemical, biological, and biochemical sensing, and the novel method of D-type fiber biosensing is a feasible means of study. PMID- 15751870 TI - Three-dimensional-object recognition by use of single-exposure on-axis digital holography. AB - On-axis phase-shifting digital holography requires recording of multiple holograms. We describe a novel real-time three-dimensional- (3-D-) object recognition system that uses single-exposure on-axis digital holography. In contrast to 3-D-object recognition by means of a conventional phase-shifting scheme that requires multiple exposures, our proposed method requires only a single digital hologram to be synthesized and used to recognize 3-D objects. A benefit of the proposed 3-D recognition method is enhanced practicality of digital holography for 3-D recognition in terms of its simplicity and greater robustness to external scene parameters such as moving targets and environmental noise factors. We show experimentally the utility of the single-exposure on-axis digital holography-based 3-D-object recognition method. PMID- 15751871 TI - Nonlocal polymerization-driven diffusion-model-based examination of the scaling law for holographic data storage. AB - For the first time to our knowledge, a detailed theoretical basis is provided for the well-known inverse-square scaling law of holographic diffraction, which states that replay diffraction efficiency eta = gamma/M2, where M is the number of gratings stored and gamma is a constant system parameter. This law is shown to hold for photopolymer recording media governed by the predictions of the nonlocal polymerization-driven diffusion model. On the basis of the analysis, we (i) propose a media inverse scaling law, (ii) relate gamma to photopolymer material parameters and the hologram geometry and replay conditions, and (iii) comment on the form and validity of the diffraction efficiency inverse-square scaling law for higher-diffraction-efficiency gratings. PMID- 15751872 TI - Compact optical roll-angle sensor with large measurement range and high sensitivity. AB - A simple and effective optical roll-angle sensor based on a magnetic garnet single crystal is demonstrated. The measurement is based on the principle that, when linearly polarized light whose polarization varies alternately in two orthogonal directions passes through a polarization analyzer, the transmitted optical intensity varies. The output voltage from an optical detector located after the analyzer can be used to determine the roll angle. The chief advantages of the sensor are that it is compact, flexible, low cost, and power effective and provides a fast response. Most importantly, the sensor is insensitive to various fluctuations such as optical intensity, photodiode responsivity, and temperature and to vibrations. This performance is most advantageous in long-distance or long term measurements. Experimental results have shown that the angular resolution is 0.01 degree in a +/-30 degree range. PMID- 15751873 TI - Wave-front reconstruction from multidirectional phase derivatives generated by multilateral shearing interferometers. AB - To increase the accuracy of wave-front evaluation, we propose to exploit the natural capability of multiple lateral shearing interferometers to measure simultaneously more than two orthogonal phase derivatives. We also describe a method, based on Fourier-transform analysis, that uses this multiple information to reconstruct the wave-front under study. PMID- 15751874 TI - Fast distance measurements by use of dynamic speckles. AB - A technique for fast measurements of the distance to an object's surface based on spatial filtering of dynamic speckle patterns is proposed. Exploitation of two spatial filters (Ronchi rulings) enables measurements to be independent of surface speed. Experimental verification of the technique is demonstrated at speeds of the surface as high as 50 m/s. The technique may find applications in machine-vision systems for acquisition of three-dimensional images and in control systems for on-line monitoring of fast-moving parts of industrial machines. PMID- 15751875 TI - Mach-Zehnder interferometer switch with a high extinction ratio over a wide wavelength range. AB - We have realized a novel Mach-Zehnder interferometer (MZI) thermo-optic switch that can operate over a much wider wavelength range than a conventional MZI switch. We constructed the novel MZI switch by incorporating a phase-generating coupler (PGC) as an optical coupler. The PGC generates a nonlinear wavelength dependent phase, which functions as a virtual wavelength-dependent delay part that cancels out the wavelength dependence of a MZI. We fabricated the novel MZI switch on a silica-based planar light-wave circuit and achieved a low insertion loss of less than 1.6 dB and a high extinction ratio of 30 dB over a wide wavelength range of 160 nm. PMID- 15751876 TI - BaTiO3 thin-film waveguide modulator with a low voltage-length product at near infrared wavelengths of 0.98 and 1.55 microm. AB - A BaTiO3 thin-film electro-optic waveguide modulator with a low half-wave voltage length product has been demonstrated at near-infrared wavelengths of 1-1.6 microm. Half-wave voltage-length products as small as 0.25 and 0.5 V cm were measured for a 5-mm-long device at wavelengths of 973 and 1561 nm, respectively. The effective electro-optic coefficients were calculated as 420 pm/V at 973 nm and 360 pm/V at 1561 nm. Further improvements in device performance by optimizing the ferroelectric domain structure are anticipated. PMID- 15751877 TI - Proposal for ideal 3-dB splitters-combiners in photonic crystals. AB - We propose a structure for ideal 3-dB splitters (1 x 2)-combiners (2 x 1) in photonic crystals. In photonic crystals, ideal 3-dB splitters based on a three port system are achievable in principle, but it is impossible to achieve ideal 3 dB combiners based on the three-port system because there is no path for a loss, owing to strong confinement of photons. The proposed structure is based on a four port system in which the concept of a microwave circuit, the so-called rat-race circuit is adopted. Design conditions for two-dimensional photonic-crystal-based rat-race circuits have been investigated by use of the coupled-mode theory in time. With the proposed structure, a 3-dB splitter-combiner has been designed. The performance of the device was numerically calculated by the finite-difference time-domain method. PMID- 15751878 TI - Emission properties of printed organic semiconductor lasers. AB - We investigated the emission properties of a distributed-feedback resonator based on an organic semiconductor patterned by a novel printing technology. We observed the peak splitting of the photonic bandstructure of the periodic grating and extracted the effective refractive index of the outcoupled guided modes. The laser works at the second diffraction order, exhibiting narrow single-mode emission at 637 nm, with a threshold as low as 37 microJ/cm2. The results suggest that direct printing is a promising fabrication technique for optically confined integrated optoelectronics. PMID- 15751879 TI - Soliton mode-locked Er:Yb:glass laser. AB - We report on a simple diode-pumped passively mode-locked Er:Yb:glass laser generating transform-limited 1536-nm solitons of 255-fs duration with a repetition rate of 50 MHz and average power of 58 mW. We also discuss timing jitter and the trade-off between short pulses and high output power in these lasers. PMID- 15751880 TI - Spectral line narrowing of a photonically generated microwave frequency comb with vertical-cavity surface-emitting laser mode-locked coupled oscillators. AB - A technique is presented for narrowing the spectral linewidth of microwave signals generated photonically by heterodyning a pair of vertical-cavity surface emitting lasers forming an extended optically coupled cavity. The experimentally demonstrated linewidth reduction, by as much as a factor of 10(4)--to less than 10 kHz in microwave frequencies up to a couple of gigahertz--is approximately ten times that expected with conventional line-narrowing techniques such as optical feedback. An interpretation is given in terms of mode locking in pairs of optically coupled lasers as a first demonstration in the frequency domain of lag synchronization of coupled oscillators. The results of theoretical modeling agree well with the experimental results. PMID- 15751881 TI - 784-nm amplified spontaneous emission from Tm3+-doped fluoride glass fiber pumped by an 1120-nm fiber laser. AB - We report 784-nm (1G4 --> 3H5 transition) amplified spontaneous emission (ASE) from Tm3+-doped fluoride (ZrF4-BaF2-LaF3-AlF3-NaF) glass fiber pumped by an 1120 nm fiber laser. To our best knowledge, this is the first report of 784-nm (1G4 - > 3H5 transition) ASE in a Tm3+-doped fluoride fiber laser. Its effects on a 480 nm (1G4 --> 3H6 transition) blue laser were also discussed. PMID- 15751882 TI - 2.1-W picosecond passively mode-locked external-cavity semiconductor laser. AB - We demonstrate an optically pumped passively mode-locked external-cavity semiconductor laser generating 4.7-ps pulses at 957 nm with as much as 2.1 W of average output power and a 4-GHz repetition rate. Compared with earlier results, the chirp of the pulses has been greatly reduced by use of an intracavity etalon. Apart from restricting the bandwidth, the etalon also helps optimize wavelength dependent gain parameters and dispersion. PMID- 15751883 TI - Second-harmonic generation in graded metallic films. AB - We study the effective second-harmonic generation (SHG) susceptibility in graded metallic films by invoking the local field effects exactly and further numerically demonstrate that graded metallic films can serve as a novel optical material for producing a broad structure in both the linear and the SHG response and an enhancement in the SHG signal. PMID- 15751884 TI - Electro-optic modulation in hybrid solgel doped with Disperse Red chromophore. AB - An electro-optically active hybrid solgel doped with Disperse Red 13 has been developed by use of a simple solvent-assisted method. It permits a high loading concentration and has low optical loss at 1550 nm. A channel waveguide amplitude modulator has been fabricated by use of active and passive hybrid solgel materials. The device shows an electro-optic coefficient of 14 pm/V at 1550 nm and stable operation. PMID- 15751886 TI - Experimental evidence of spontaneous symmetry breaking in intracavity type II second-harmonic generation with triple resonance. AB - We describe a spontaneous symmetry-breaking phenomenon between the intensities of the ordinary and extraordinary components of the fundamental field in intracavity type II harmonic generation. It is based on a triply resonant cavity containing a type II chi(2) crystal pumped at fundamental frequency omega0. The pump beam generates a second-harmonic mode at frequency 2omega0 that acts as a pump for frequency-degenerate type II parametric downconversion. Under operating conditions symmetric with respect to the ordinary and extraordinary components of the fundamental wave, we show a breaking of the symmetry of the intensities of these two waves. PMID- 15751885 TI - Three-element phased-array approach to diffuse optical imaging based on postprocessing of continuous-wave data. AB - We present a multielement phased-array approach to diffuse optical imaging based on postprocessing of continuous-wave data for the improvement of spatial resolution. In particular, we present a theoretical and experimental analysis of the performance of a three-element source array in the study of an optically turbid medium with two embedded cylindrical inclusions. We find that the proposed phased-array approach is able to resolve two cylinders with side-to-side separation of 10 mm that are not resolved by the intensity associated with a single light source. PMID- 15751887 TI - Ultrasensitive femtosecond two-photon detector with resonantly enhanced nonlinear absorption. AB - We report on two-photon detection based on nonlinear absorption between subbands in quantum wells. Resonantly enhanced nonlinear absorption, 6 orders of magnitude higher than that of typical bulk semiconductors, leads to a threshold power density for quadratic detection as low as 0.1 W/cm2 and to femtosecond time resolution. The approach facilitates dynamic characterization of the optical light field of infrared emitters with unprecedented sensitivity. We also present a new method for determining the intersubband and intrasubband scattering times by means of autocorrelation measurements. PMID- 15751888 TI - Optical parametric oscillator with a pulse repetition rate of 39 GHz and 2.1-W signal average output power in the spectral region near 1.5 microm. AB - We present a singly resonant, synchronously pumped optical parametric oscillator with a record-high repetition rate of 39 GHz. The transform-limited 2.2-ps pulses at 1570 nm have as much as 2.1-W average output power. The all-solid-state pump source is based on a diode-pumped passively mode-locked 39-GHz Nd:YVO4 laser and an efficient ytterbium-doped fiber amplifier. PMID- 15751889 TI - Generation of pulsed polarization-entangled photon pairs in a 1.55-microm band with a periodically poled lithium niobate waveguide and an orthogonal polarization delay circuit. AB - We report a scheme for generating pulsed polarization-entangled photon pairs based on conversion from time-bin entanglement to polarization entanglement by use of an orthogonal polarization delay circuit and post-selection. We have experimentally demonstrated the scheme, using a periodically poled lithium niobate waveguide, and successfully obtained polarization entanglement in the 1.55-microm telecom wavelength band. PMID- 15751890 TI - Optimal nonmonotonic convergence of the iterative Fourier-transform algorithm. AB - The increase of the monotonic convergence rate is an important issue for iterative Fourier-transform algorithms. However, the steepest monotonic convergence of the iterative Fourier-transform algorithm does not always promise an optimal solution in the design of a diffractive optical element. The optimal nonmonotonic convergence of the iterative Fourier-transform algorithm is investigated by employing a microgenetic algorithm. The proposed hybrid scheme of the iterative Fourier-transform algorithm and the microgenetic algorithm show nonmonotonic convergence, and this results in a superior design. PMID- 15751891 TI - High-resolution optical storage by use of minute spheres. AB - We have developed a new optical storage system that uses minute spheres arranged upon a surface-relief grating. By using dye-doped minute spheres as recording bits we can limit a sensitive region within a sphere's diameter. Using a reflection-type confocal optical system, we read out shape signals from minute spheres at high resolution. The shape signals from minute spheres are utilized as clock signals in recording and readout. PMID- 15751892 TI - Photonic crystal device sensitivity analysis with Wannier basis gradients. AB - We present a powerful sensitivity analysis method for devices in a photonic crystal. The method is based on a Wannier basis field expansion and efficient matrix analysis techniques for finding eigenvalue and transmission gradients with respect to the perturbation. The method permits fast analysis of a large number of dielectric perturbation situations for multiple devices in a photonic crystal. We verify the method with finite-difference time-domain and plane-wave expansion calculations. PMID- 15751893 TI - One-beam recording in a LiNbO3 crystal. AB - We propose a new way to record images in a photorefractive LiNbO3 crystal. This method involves only a single object light without any reference light. We believe that the recording is attained by fanning holograms that result from interference between the object light and its scattered light. Although volume gratings are involved, the recorded pattern can be viewed not only with a laser light beam that is incident over a certain angular range but also with white light. PMID- 15751894 TI - Polarization properties of vectorial nonparaxial Gaussian beams in the far field. AB - The polarization properties of vectorial coherent nonparaxial Gaussian beams are studied. It is shown that, when the source of a nonparaxial Gaussian beam is completely polarized, the degree of polarization of the propagation field maintains a constant value of 1. However, when the source is completely unpolarized, the degree of polarization does not maintain a constant value of 0. In the far field, vectorial nonparaxial Gaussian beams can become partially polarized, except on the propagation axis, and are spatially nonuniform across a typical cross section of the beam even though the source field is completely unpolarized. PMID- 15751895 TI - Fully quantized theory of four-wave mixing with bosonic matter waves. AB - We solve exactly a set of fully quantized coupled equations that describe the quantum dynamics, including the evolution of quantum fluctuations, entanglements, and correlations, of four-wave mixing (FWM) with matter waves. The analytical solution reveals Rabi oscillations, collapses and revivals, and agrees with FWM experiments. It also applies to optical FWM and the Rayleigh superradiance in Bose-Einstein condensates. PMID- 15751896 TI - Engineering the frequency correlations of entangled two-photon states by achromatic phase matching. AB - We put forward a new method to control the spectra of photon pairs generated in parametric downconversion that allows their spectral properties to be tuned from correlation to anticorrelation, including uncorrelation. The method employs tilted pulses and can be implemented in materials and frequency bands for which conventional methods do not hold. PMID- 15751897 TI - Ultrabright femtosecond source of biphotons based on a spatial mode inverter. AB - A method of enhancing the efficiency of entangled biphoton sources based on a type II femtosecond spontaneous parametric downconversion (SPDC) process is proposed and implemented experimentally. Enhancement is obtained by mode inversion of one of the SPDC output beams, which allows the beams to overlap completely, thus maximizing the number of SPDC photon pairs with optimum spatiotemporal overlap. By use of this method, biphoton count rates as high as 16 kHz from a single 0.5-mm-long beta-barium borate crystal pumped by second harmonic radiation from a Ti:sapphire laser were obtained. PMID- 15751898 TI - Measurement of the collision time of dense electronic plasma induced by a femtosecond laser in fused silica. AB - Electronic plasma induced by a focused femtosecond pulse (130 fs, 800 nm) in fused silica was investigated by use of pump-probe technology. Pump and probe shadow imaging and interferometric fringe imaging were combined to determine electronic collision time tau in the conduction band, and tau was measured to be 1.7 fs at an electron density near 5 x 10(19) cm(-3). The lifetime of the electronic plasma is also measured to be approximately 170 fs by use of the time resolved shadow imaging technique. PMID- 15751899 TI - Diffraction-based femtosecond pulse shaping with a two-dimensional spatial light modulator. AB - A new diffraction-based method is proposed and demonstrated for simultaneous shaping of both the phase and amplitude of femtosecond laser pulses by use of a phase-only two-dimensional spatial light modulator. The method suppresses certain types of temporal replica features ordinarily observed in femtosecond pulse shaping owing to imperfections in modulator devices and allows for multiplexed outputs suitable for use in various applications. PMID- 15751900 TI - Interferometric technique for measuring broadband ultrashort pulses at the sampling limit. AB - We present a new technique for measuring ultrashort optical pulses by use of spectral phase interferometry for direct electric-field reconstruction that is suitable for large bandwidth pulses. The method does not require generation of a replica of the pulse to be measured and encodes the spectral phase information in a spatial interference pattern. A major advantage of this method is that the spectral sampling saturates the Whittaker-Shannon bound. Moreover, the technique allows for the characterization of some types of space-time coupling. An experimental demonstration of the technique is presented. PMID- 15751901 TI - 220-fs erbium-ytterbium:glass laser mode locked by a broadband low-loss silicon/germanium saturable absorber. AB - We demonstrate femtosecond performance of an ultrabroadband high-index-contrast saturable Bragg reflector consisting of a silicon/silicon dioxide/germanium structure that is fully compatible with CMOS processing. This device offers a reflectivity bandwidth of over 700 nm and subpicosecond recovery time of the saturable loss. It is used to achieve mode locking of an Er-Yb:glass laser centered at 1540 nm, generating 220-fs pulses, with what is to our knowledge the broadest output spectrum to date. PMID- 15751902 TI - Monolithic carrier-envelope phase-stabilization scheme. AB - A new scheme for stabilizing the carrier-envelope (CE) phase of a few-cycle laser pulse train is demonstrated. Self-phase modulation and difference-frequency generation in a single periodically poled lithium niobate crystal that transmits the main laser beam allows CE phase locking directly in the usable output. The monolithic scheme obviates the need for splitting off a fraction of the laser output for CE phase control, coupling into microstructured fiber, and separation and recombination of spectral components. As a consequence, the output yields 6 fs, 800-nm pulses with an unprecedented degree of short- and long-term reproducibility of the electric field waveform. PMID- 15751903 TI - Special issue: obesity in Louisiana. PMID- 15751904 TI - The obesity epidemic: incidence and prevalence. AB - Obesity is a major public health problem in the United States and in Louisiana in particular. The prevalence of both overweight and obesity has increased in the past two decades. National studies and studies conducted in Louisiana in adults and children have shown that overweight and obesity are related to conditions such as hypertension, diabetes, stroke, and insulin resistance syndrome. Obesity tracks over time, with obese children and adolescents being more likely to become obese adults. Numerous intervention models to modify eating and exercise behaviors have been conducted with children in Louisiana and have shown that it is possible to reduce fat and saturated fat intakes and increase the amount of moderate to vigorous daily physical activity. More research is needed, however, in both health education programs and environmental influences if we are to be effective in making the necessary changes in eating and physical activity habits of children, adolescents, and adults to reduce the prevalence of obesity. PMID- 15751905 TI - Genetic and physiological factors in obesity. AB - Body weight is determined by the interaction of the genetic makeup of an individual and the environment in which that person is living. The control systems that regulate body weight are numerous and include signals from fat that travel to the hypothalamus where cognitive and internal signals are integrated. The integration of these signals involves a complex array of neuropeptides, neurotransmitters and structural circuits. These circuits regulate appetite, intake and energy expenditure. Recent studies demonstrate that the theory of a thrifty genotype is probably correct. Some people are more susceptible to our obesogenic environment than others. Some people are able to overwhelm their genetics by voluntarily increasing energy expenditure and decreasing food intake; a feat that is rarely accomplished and requires a Herculean effort. As we better understand the environmental, genetic, physiological, and behavioral aspects of obesity, we will undoubtedly develop better strategies and therapies for obesity. PMID- 15751906 TI - Environmental and lifestyle influences on obesity. AB - Obesity is an end result of the intricate interactions of biology, behavior, and environment. Recent hypotheses in the scientific community suggest the current obesity epidemic is being driven largely by environmental factors (e.g., high energy/high fat foods, fast food consumption, television watching, "super-sized" portions, etc.) rather than biological ones. Individuals are bombarded with images and offers of high fat, high calorie, highly palatable, convenient, and inexpensive foods. These foods are packaged in portion sizes that far exceed federal recommendations. Furthermore, the physical demands of our society have changed resulting in an imbalance in energy intake and expenditure. Today's stressful lifestyles compound the effects of environmental factors by impairing weight loss efforts and by promoting fat storage. Combating the obesity epidemic demands environmental and social policy changes, particularly in the areas of portion size, availability of healthful foods, and promotion of physical activity. PMID- 15751907 TI - Politics of the obesity epidemic: results of the 2003 Louisiana Physicians Survey. AB - The obesity epidemic is producing growing interest in public policy measures targeting prevention. We developed a printed survey suitable for mailing to primary care physicians in Louisiana. Questions were asked concerning health policies relating to motorcycle helmet use, smoking, and obesity. We also obtained personal data, including type of practice, gender, age, ethnicity, personal income, height, and weight. Of 993 surveys mailed, there were 218 responses: 74% were male, 55% were from East Baton Rouge parish, an urban location, and 84% had an income of > dollar 100,000. Of respondents, 74% strongly agreed with helmet laws, 73% strongly agreed with limitations on smoking spaces and 62% strongly agreed with cigarette taxation. Concerning health policies related to obesity prevention, strongest support was for school concession policies, with 73% reporting strong agreement or agreement. There was support for regulating food advertisements, where 60% agree or strongly agree. However, only 29% agree or strongly agree with the taxing of unhealthy foods, compared to 78% of respondents favoring cigarette taxation, a statistically significant difference (< 0.0001). Support was particularly weak for governmental involvement in obesity discrimination, with only 16% of respondents agreeing or strongly agreeing. Political response to the obesity epidemic is only now emerging. However, our survey indicates that support among Louisiana physicians for obesity related policy is not equal to support for policies directed against helmet and smoking laws. PMID- 15751908 TI - Can obesity prevention work for our children? AB - The prevalence of obesity in children and adolescents is higher than 20 years ago in all racial-ethnic, age, and gender groups. Research has lead to the discovery of many risk factors for obesity, which may help practitioners target at-risk individuals. Insight concerning obesity prevention can come from examining other public health programs, which center on prevention; such as smoking, seat belt use, and sexually transmitted disease. Another guide when establishing obesity prevention is evaluation of currently successful programs. Prevention and treatment interventions for childhood obesity should promote the replacement of unhealthy eating and exercise practices with healthier behaviors. The goal of prevention should always be maintenance of normal growth patterns, rather than weight loss. In predisposed children, sedentary, non-nutritious environments challenge metabolic capacity and promote overweight conditions, further inactivity and increased sedentary behaviors. This results in clinically significant obesity, reduced insulin sensitivity and ultimately type 2 diabetes later in life. Prevention of future chronic disease in children and adults may depend on our ability to prevent the onset of obesity in young children. This should be a primary goal of pediatricians, family health care professionals, and public health professionals. PMID- 15751909 TI - Obesity related morbidity and mortality. AB - The epidemic of obesity has highlighted the extent of the risks associated with this disease. The risks arise from the increased mass of fat tissue, as well as the products produced by the increased number and size of fat cells in obese individuals. Psychosocial dysfunction, obstructive sleep apnea, and osteoarthritis can be a direct result of increased fat mass. Other diseases associated with obesity result from the metabolic consequences of enlarged fat cells. Diabetes, gallbladder stones, high blood pressure, liver disease, coronary artery disease, cerebrovascular disease, certain types of cancers, and infertility can all be traced in part to the increased secretion of inflammatory and coagulation molecules from fat cells. Finally, obesity also increases overall mortality. It is clear from this review that the morbidity and increased mortality of overweight and obesity is substantial and should prompt further attention towards the need for appropriate weight management in health care. PMID- 15751910 TI - Behavior and lifestyle: approaches to treatment of obesity. AB - The increasing prevalence of overweight and obesity in adults and children demonstrates a steadily growing epidemic. This rising rate of obesity is associated with obesity related comorbidities including cardiovascular disease, hypertension, some cancers, joint disease, and particularly, type 2 diabetes. Modest weight loss (5% to 10% of total body weight) through lifestyle intervention approaches has been found to have a beneficial effect on comorbid conditions, particularly hypertension and type 2 diabetes. Effective behavioral treatment of obesity involves modification of eating and physical activity patterns to yield negative energy balance. Research studies have found that interventions that combine a low-calorie diet, increased physical activity, and behavior therapy are most effective for weight loss and maintenance. Furthermore, extended length of treatment contact, weight loss satisfaction, and social support may promote positive long-term outcomes in obese adults and children. PMID- 15751911 TI - Pharmacologic therapy for obesity. AB - Obesity is a chronic condition, and long-term treatment will most likely be needed. Approved prescription medications for weight loss appear to have similar efficacy in controlled studies. No predictors of responsiveness in an individual patient or class of patients have been established. The choice of a medication is based on the underlying medical indication or contraindication of a particular drug, concurrent medication, age of the patient, need for monitoring, anticipation of the length of therapy, and the preference of a patient. Behavioral and dieting interventions, and increased physical activity are considered the primary means to promote and maintain weight loss. Weight-loss medications should be considered only as an adjunct for patients who are at substantial risk because of their obesity and in whom non-pharmacologic treatments have not resulted in sufficient weight loss to improve health or to prevent weight regain. PMID- 15751912 TI - Bariatric surgery. AB - The surgical treatment of obesity is indicated in patients who have failed sincere attempts at medical therapies to lose weight. The BMI must exceed 40 or exceed 35 and be associated with at least two comorbid conditions. Comorbid conditions include diabetes mellitus, hypertension, obstructive sleep apnea, hyperlipidemia, and other weight related conditions that may benefit from weight loss. Patients need to be educated concerning the specific operation to be performed. They must be taught what they need to do to optimize the likelihood of success of the surgery, and they must have an understanding concerning the potential adverse side effects. When this format is followed, bariatric surgery is a legitimate treatment for morbid obesity, and the only treatment that is generally successful. PMID- 15751913 TI - The burden of communicable diseases. PMID- 15751915 TI - Prevalence and distribution of anopheline mosquitoes in malaria endemic areas of Asir region, Saudi Arabia. AB - To study the prevalence of anopheline mosquitoes, over 180 sites were sampled in malaria-endemic areas of Asir region, Saudi Arabia, during June 1999-April 2001. A total of 7085 larval and 754 adult female Anopheles spp. specimens were collected. Seven species were identified: An. dthali, An. rupicolus, An. sergentii, An. arabiensis, An. multicolor, An. turkhudi and An. pretoriensis. Both An. arabiensis and An. sergentii are known vectors of malaria in the region. An. dthali occurred in all sites and was the most abundant species. An. turkhudi was collected in low numbers as larvae only. An. multicolor and An. pretoriensis were recorded for the first time in Asir region. An. sergentii is a species of the northern areas of the region, whereas An. arabiensis was more prevalent in the south. PMID- 15751914 TI - High altitude epidemic malaria in Bamian province, central Afghanistan. AB - We report an epidemic of Plasmodium falciparum malaria in the remote valley of Bamian (altitude 2250 m-2400 m) in the central highlands of Afghanistan. A team of malaria experts from the World Health Organization and HealthNet International carried out the investigation. A total of 215 peripheral blood smears were obtained and 63 cases of malaria (90.5% P. falciparum, the remainder P. vivax) were confirmed. The study revealed that areas vulnerable to malaria in Afghanistan are more widespread than previously recognized. The area had been malaria-free until recently, when the disease appears to have been introduced as a consequence of protracted conflict and resultant population movement, and transmitted locally during the short summer months. The outbreak led to severe morbidity and high mortality in a province having only a few poorly-provisioned health care facilities. PMID- 15751916 TI - Knowledge, perceptions and prevention of malaria among women in Sistan va Baluchestan, Islamic Republic of Iran. AB - A questionnaire about malaria knowledge, beliefs and practices was given to a random sample of 2168 married women from rural and urban areas of Sistan va Baluchestan, Islamic Republic of Iran. The mean knowledge score of subjects was low at 5.5 (maximum 15.0). Few respondents (37.6%) knew that malaria was an important disease in the area and only 58.4% knew that malaria was transmitted by mosquitoes. Most subjects (69.4%) never used a mosquito net. Only 49.9% of rural and 73.8% of urban residents would seek care for fever and chills from the local health centre. Community health workers (behvarz) were the main source of information (29.5%) for rural women; the role of physicians in education was minimal. Subsequent health education must be tailored to the educational needs of the target population in this area. PMID- 15751917 TI - Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran. AB - A species-specific polymerase chain reaction (PCR) assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran. PMID- 15751918 TI - Seroprevalence of three emerging arboviral infections in Kuwaiti nationals. AB - Diseases caused by dengue, sandfly fever and hanta viruses pose a major health risk in many countries. We determined the threat of these arboviral infections through a serologic using enzyme linked immunosorbent assay (ELISA) based tests. Hantavirus-specific antibodies were also detected using immunofluorescence. Of 499 samples tested for dengue virus IgG antibodies l4% were as positive for dengue positive by all the ELISA tests. Among the 42 showing strong IgG reactivity, only 1 was positive for dengue virus IgM antibodies. All samples tested for IgG antibodies to sandfly fever virus were negative. Hantavirus antibodies were detected in 11% of the 46 samples from high-risk individuals. The low prevalences suggest that at present these infections are not a serious problem in Kuwait. PMID- 15751919 TI - Removing hepatitis C virus from polytetrafluoroethylene-coated orthodontic archwires and other dental instruments. AB - The efficacy of removing, rather than destroying, hepatitis C virus (HCV) from polytetrafluoroethylene (PTFE)-coated orthodontic archwirs was tested. PTFE coated archwires, pieces of PTFE and endodontic files (20 each) were immersed in HCV-infected blood for 24 hours. Half were washed under running water for 10 seconds while the remainder were thoroughly scrubbed with a toothbrush under running water for 10 seconds. Items were kept in separate dishes of distilled water for 24 hours. Reverse transcription polymerase chain reaction was used for viral replication. Viruses were detected by the gel method using a transilluminator. Only scrubbed PTFE pieces and coated archwires were negative for HCV. This suggests that PTFE coating of dental instruments inhibited HCV adhesion when thoroughly scrubbed. PMID- 15751920 TI - Mycoplasma pneumoniae infection in Yemen: incidence, presentation and antibiotic susceptibility. AB - To determine the incidence, clinical presentation and antibiotic susceptibility of Mycoplasma pneumoniae at the main hospitals in Sana'a, we studied 405 patients clinically and radiographically diagnosed with lower respiratory tract infections aged 10-60 years. M. pneumoniae was identified by 3 different methods: culture, antigen detection and IgM serology. Antibiotic susceptibility testing was performed for confirmed isolates by macro-broth dilution technique. There were 125 patients (30.9%) with current infection, mostly among younger age groups, with bronchopneumonia the most common underlying clinical condition. All tested isolates were susceptible to all antibiotics in the in vitro antibiogram, with erythromycin the most active. The results indicate the need for different approaches in the diagnosis of M. pneumoniae infection in Yemen. PMID- 15751921 TI - Prevalence of Ureaplasma urealyticum and Mycoplasma genitalium in men with non gonococcal urethritis. AB - This study in Teheran, Islamic Republic of Iran, investigated the prevalence of Ureaplasma urealyticum and Mycoplasma species in men with non-gonococcal urethritis. Urethral swab samples were collected from 125 cases and 125 healthy men as a control group. The samples were then investigated by culture methods. The rates of detected bacteria in case and control groups were 19.2% and 7.2% for U. urealyticum, 7.2% and 0.8% for M. genitalium, and 2.4% and 1.6% for M. hominis respectively. Statistical analysis showed a significant difference between case and control groups in the prevalence of U. urealyticum and M. genitalium but not M. hominis. It is concluded that in men, U. urealyticum and M. genitalium may have an etiologic role in non-gonococcal urethritis. PMID- 15751922 TI - Women's perceptions and practices regarding their rights to reproductive health. AB - ABSTRACT At the outpatient clinic of Ain Shams University Maternity Hospital, perceptions and practices of 1000 women regarding their reproductive health rights (reproductive rights) were evaluated. The majority had positive perceptions about their rights to reproductive health; however, 30% disagreed with prohibitions of discrimination against women, particularly prohibitions of female genital mutilation (FGM). A significant association was found between perceptions of sexual rights and demographic characteristics and between education and practices regarding early detection of cancer. No significant association was found between education and concepts of adolescent health education as a reproductive right. Programmes about women's reproductive rights that emphasize the issue of sexual health through religious education are recommended as one of the best strategies for the eradication of FGM. PMID- 15751923 TI - Induced labour with prostaglandin E2 in different parity groups after premature rupture of membranes. AB - The study compared the outcome of induction of labour with prostaglandin E2 vaginal tablets in patients with premature rupture of membranes (PROM) at term in different parity groups. A retrospective review was made of the hospital records of 169 women attending the maternity unit of King Faisal Military Hospital, Saudi Arabia. There were no statistically significant differences between the 3 groups (parity 0, parity 1-4 and parity 5+) in rates of labour augmentation, caesarean sections, neonatal intensive care admissions or low Apgar scores. There were no serious complications of induction of labour such as infection or uterine hyperstimulation or rupture. Prostaglandin E2 may be used with care for labour induction in women with PROM at term, even grand multiparas, unless there is history of previous caesarean delivery. PMID- 15751924 TI - Outcome of induced labour in pregnancies at 41 weeks gestation and over in Saudi Arabia. AB - A retrospective, descriptive cohort study was conducted at King Faisal Military Hospital, Saudi Arabia, to compare pregnancy outcomes in patients induced with prostaglandin E2 from 41 weeks gestation. A total of 450 women whose antenatal care and delivery were conducted at the hospital during 1995-99 were studied. The main outcome measures used were caesarean section rate and perinatal morbidity and mortality. In otherwise normal pregnancies, the caesarean section rate was not significantly increased when induction of labour was carried out at 41 weeks gestation compared with > or =42 weeks. Although more perinatal complications occurred when induction was carried out at 42 weeks, the results were not statistically significant. A large prospective clinical trial is indicated. PMID- 15751925 TI - United Arab Emirates National Newborn Screening Programme: an evaluation 1998 2000. AB - To evaluate the United Arab Emirate National Newborn Screening Programme we compared coverage, timeliness of programme indicators (age at sampling, recall and treatment initiation, timing of specimen delivery and laboratory results) and specimen quality with international standards. Recall rate, sensitivity, specificity, positive predictive values and relative incidence rates for phenylketonuria (PKU) and congenital hypothyroidism (CH) were calculated. Investigations for hypothyroidism included thyroid function studies (T3, T4, fT4 and TSH), technetium-99m thyroid scan when possible and thyroglobulin and thyroid antibodies when indicated. PKU investigations included plasma amino acids and measurement of biopterin defects. In the 6 years before December 2000, 138,718 neonates were screened. Relative incidences for CH and for classic PKU were 1:1570 and 1:20,050 respectively. PMID- 15751926 TI - Study of mortality risk factors for children under age 5 in Abu Dhabi. AB - We investigated the association of biological, sociocultural and economic risk factors with child mortality in Abu Dhabi from 1 January-31 December 1997. With McNemar chi-squared test, most selected biological risk factors were statistically associated with child mortality, although maternal age older than 40 years and history of fetal death were not positively correlated with neonate, infant or age under 5 mortality. Among sociocultural and economic risk factors, maternal lack of formal education and low monthly income were significantly associated with child death. Consanguinity was significantly associated with under 5 and infant but not neonatal mortality. Gestation <37 weeks was highly associated with mortality among all ages. Strengthening health care programmes and emphasizing the need to identify high risk groups should be priorities. PMID- 15751927 TI - Student screening for inherited blood disorders in Bahrain. AB - In Bahrain and neighbouring countries inherited disorders of haemoglobin, i.e. sickle-cell disease, thalassaemias and glucose-6-phosphate dehydrogenase (G6PD) deficiency, are common. As part of the National Student Screening Project to determine the prevalence of genetic blood disorders and raise awareness among young Bahrainis, we screened 11th-grade students from 38 schools (5685 students), organized lectures and distributed information about these disorders. Haemoglobin electrophoresis, high performance liquid chromatography, blood grouping and G6PD deficiency testing were performed. Prevalences were: 1.2% sickle-cell disease; 13.8% sickle-cell trait; 0.09% beta-thalassaemia; 2.9% beta-thalassaemia trait; 23.2% G6PD deficiency; 1.9% G6PD deficiency carrier. Health education, carrier screening and premarital counselling remain the best ways to reduce disease incidence with potentially significant financial savings and social and health benefits. PMID- 15751928 TI - [Knowledge and practice of doctors and midwives working in primary health care regarding screening for cervical and breast cancers]. AB - We assessed the knowledge and practices of breast and cervical cancer of 286 physicians and 126 midwives working in primary health care in Tunis who responded to an anonymous questionnaire. Questions were related to knowledge of the epidemiology and survival rates in the early stages of breast and cervical cancers, to training in this domain and to the degree of involvement in this screening. The knowledge of the two cancers was relatively modest among both physicians and midwives. The systematic practice of Pap smear was significantly more frequent among midwives than physicians. The same result was observed for systematic clinical breast examination. Lack of training about carrying out Pap smears and the large number of consultations were the main factors negatively associated with systematic Pap smear and clinical breast examination practice. PMID- 15751930 TI - Oesophageal cancer in Bahrain. AB - ABSTRACT Data from all 148 cases of oesophageal cancer in Bahrain during 1952-99 were analysed according to patient sex and age and site and histological pattern of tumour, and compared with other Gulf countries. In Bahrain, oesophageal cancer accounted for 2.6% of malignant neoplasms. The female:male ratio was 1.8:1, and the majority of patients were > or =51 years and < or =70 years of age. The lower and upper third of the oesophagus were the most and least frequently involved sites, respectively. Squamous carcinoma (males) and adenocarcinoma (females) were the main histological types. There is varying consistency between these data and those of other Gulf countries having similar anthropological and demographic profiles. A prospective study may help to better understand the aetiology of the disease and inform preventive policies. PMID- 15751929 TI - Epstein-Barr virus in head and neck squamous cell carcinoma. AB - We compared 50 patients with head and neck squamous cell carcinomas (cases) and 45 matched healthy controls. Biopsy specimens were taken from tumours and normal tissue of the cases and controls respectively and serial paraffin embedded sections were processed to detect Epstein-Barr (EB) viral antigen. We found EB viral proteins in 38% of cases and none in controls, which suggests a positive correlation. Serum samples were also tested for the presence of EB virus IgG by ELISA for comparison with immunohistochemical findings. Patients with positive immunohistochemical staining results had significantly higher mean antibody titres compared with those with negative results. ELISA may be useful in determining the etiology of head and neck cancers, but the results are not unequivocally reliable. PMID- 15751931 TI - Distribution and correlates of total impaired fasting glucose in Oman. AB - To determine the distribution and correlates of total impaired fasting glucose in Oman, we conducted a cross-sectional survey of 1968 households with 7011 eligible residents in 2000. During face-to-face interviews, demographic data, blood pressure, fasting blood glucose, serum cholesterol, weight, height and waist and hip measurements were obtained. Of 5788 subjects tested for total impaired fasting glucose (response rate: 83%), crude prevalence was 17.3% and age-adjusted prevalence was 20.3%. Associated factors in bivariate analysis were older age, male gender, lower education, hypercholesterolaemia, being married, obesity, abnormal waist to hip ratio and hypertension. Total impaired fasting glucose is a significant public health problem in Oman. Increasing the awareness of health care providers and community through health education is crucial. PMID- 15751932 TI - Comparative study of lung function in Iranian factory workers exposed to silica dust. AB - The study compared lung function among 322 workers in pottery, ceramic, stone cutter and stone-grinder factories in the west of the Islamic Republic of Iran. Concentrations of silica particles <2 microm were measured in the ambient air of factories. Forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC) were significantly lower in stone-grinders compared with pottery, ceramic or stone-cutter workers and a control group. No difference in lung function was found in pottery and stone-cutter workers with less than 20 years occupation compared with controls. The prevalence of respiratory symptoms in stone-grinders was higher than other workers. The concentration of silica particles of stone grinder factories was 40-110 times higher than in ceramic and potteries factories. More attention is needed to ventilation systems and health care of stone-grinders. PMID- 15751933 TI - Possible new role for angiotensin-converting enzyme inhibitors in treating glomerulonephritis. AB - Serum transforming growth factor-beta (TGF-beta1) production was estimated for 10 patients with essential hypertension, 12 patients with glomerulonephritis (5 hypertensive and 7 normotensive) and 10 healthy controls. The glomerulonephritis group received angiotensin-converting enzyme inhibitor captopril 25-75 mg/day for 4 weeks. Blood urea, serum creatinine, 24-hour urinary protein and serum TGF beta1 were then re-estimated. Urea and creatinine were significantly higher in the hypertension and glomerulonephritis groups than in the controls and also higher in the glomerulonephritis group than the hypertension group. TGF-beta1 was significantly higher in the glomerulonephritis groups than in the control and hypertension groups. TGF-beta1 and 24-hour urinary protein were significantly reduced in the glomerulonephritis group. PMID- 15751934 TI - Evaluation of 2-column agglutination versus conventional tube technique for antibody screening. AB - The study aimed to determine the specificity and sensitivity of the Ortho BioVue two-column agglutination system for the detection of low concentrations of clinically significant antibodies in serum. The BioVue system was compared with the conventional tube technique (LISS-Coombs indirect antiglobulin test), and the two-stage Papenzyme test was used to resolve discrepancies between the two methods. We tested 3000 serum samples from randomly selected patients at King Hussein Medical Centre. Both the antibody screening and identification gave negative results in 2952 patients and positive results in 48 patients. We found the BioVue system to be the more sensitive technique. However, if papain enzyme treated cells were included in the conventional tube technique when applied to antibody screening and identification, both methods would be of comparable sensitivity. PMID- 15751935 TI - Histopathology of human intestinal anastomosis. AB - The faultless seal of the anastomosis is an important aspect of abdominal surgery but opportunities to make a histological evaluation of human intestinal anastomosis sites are rare. This retrospective study examined 30 anastomoses that had been resected following complications or postmortem; the aim was to describe the histological changes at various stages of healing ranging from 4 days to 3.5 years post-surgery. Anastomosis dehiscence showed features of extensive mucosal necrosis and bad submucosal apposition. Old healed anastomoses continued to have chronic inflammatory cells and muscular discontinuity with areas of intervening fibrosis. It would be useful to understand more about how the intraluminal contents are propelled through this scarred area. PMID- 15751936 TI - Patient satisfaction with primary health care services in two districts in Lower and Upper Egypt. AB - This study compares patient satisfaction with primary health care services and identifies factors associated with patient satisfaction in two health districts in Egypt where a project for upgrading primary health care services had been running for three years. An exit interview was conducted for 1108 patients using a structured questionnaire. The results revealed that most clients using primary health care services were females. Patient satisfaction was high for accessibility, waiting area conditions and performance of doctors and nurses. The main complaints centred on the availability of prescribed drugs and laboratory investigations. Additionally, level of privacy in the consultation room was described as unsatisfactory by 33% of patients. There was no association between overall patient satisfaction and age, gender, education level or type of service received. PMID- 15751937 TI - [Survey of surgical and radiological semiology modules and their adaptation to the Arabic cultural environment]. AB - Many professors of medicine oppose the Arabization of the teaching of medical science in the Maghreb countries, under pretext of the inability of Arabic to convey scientific concepts objectively compared with French. We made a qualitative survey of surgical and radiological semiology used in the Faculty of Medecine, Ibn El Jazzar. Terms, expressions and synonyms were identified and classified according to their cultural load into two categories: "culturally adapted terms" and "culturally strange terms" in relation to the national culture. It was evident that the majority of the recorded expression were based on Western culture. Thus the hypothesis of the neutrality of the French language in the medical teaching is invalid. Furthermore the use of French poses difficulties for students in understanding the scientific matter taught, and indirectly promotes Western culture within the medical academic establishments of the Arabic world. The use of the mother tongue in teaching medicine is today an educational necessity. PMID- 15751938 TI - Effects of passive smoking on children's health: a review. AB - Since the mid-1980s there has been increasing interest in the effects of passive smoking on the health of children. It has been estimated that the total nicotine dose received by children whose parents smoke is equivalent to their actively smoking between 60 and 150 cigarettes per year. This review article considers the evidence for a relationship between passive smoking and disorders such as: prenatal damage to the fetus; poor growth indicators; respiratory illness; atopy and asthma; coronary heart disease; and sudden infant death syndrome. We conclude that paediatricians should not be complacent about the hazards of passive smoking for children and that public health education efforts should be continued. PMID- 15751940 TI - Community context of health system development: implications for health sector reform in Pakistan. AB - To describe local sociopolitical and organizational factors that influence health system development in Karachi, Pakistan, we conducted participant observation while working with health providers and communities in one urban district to reorient services towards a primary health care district health system. We found that the community characteristics, particularly the diverse socialpolitical and cultural make-up and organizational complexity that involved multiple levels of government, influenced efforts towards collaboration and shaped the development of the health system. We conclude that for effective implementation of health sector reform there is a need to comprehend fully the community context and complexity of existing health service provision. PMID- 15751939 TI - Breast cancer in Egypt: a review of disease presentation and detection strategies. AB - Carcinoma of the breast is the most prevalent cancer among Egyptian women and constitutes 29% of National Cancer Institute cases. Median age at diagnosis is one decade younger than in countries of Europe and North America and most patients are premenopausal. Tumours are relatively advanced at presentation. The majority of tumours are invasive duct subtype and the profile of hormone receptors is positive for estrogen receptors and/or progesterone receptors in less than half of cases. This overview examines genetic changes, potential and established predictive and prognostic markers and end results of surgery, radiotherapy and systemic therapy for early, locally advanced and metastatic disease stages. Disease presentations common to the region and early detection strategies are presented. PMID- 15751941 TI - Factor V Leiden and fetal loss in a 33-year-old Tunisian woman. PMID- 15751942 TI - Photocycle of the flavin-binding photoreceptor AppA, a bacterial transcriptional antirepressor of photosynthesis genes. AB - The flavoprotein AppA from Rhodobacter sphaeroides contains an N-terminal domain belonging to a new class of photoreceptors designated BLUF domains. AppA was shown to control photosynthesis gene expression in response to blue light and oxygen tension. We have investigated the photocycle of the AppA BLUF domain by ultrafast fluorescence, femtosecond transient absorption, and nanosecond flash photolysis spectroscopy. Time-resolved fluorescence experiments revealed four components of flavin adenine dinucleotide (FAD) excited-state decay, with lifetimes of 25 ps, 150 ps, 670 ps, and 3.8 ns. Ultrafast transient absorption spectroscopy revealed rapid internal conversion and vibrational cooling processes on excited FAD with time constants of 250 fs and 1.2 ps, and a multiexponential decay with effective time constants of 90 ps, 590 ps, and 2.7 ns. Concomitant with the decay of excited FAD, the rise of a species with a narrow absorption difference band near 495 nm was detected which spectrally resembles the long living signaling state of AppA. Consistent with these results, the nanosecond flash-photolysis measurements indicated that formation of the signaling state was complete within the time resolution of 10 ns. No further changes were detected up to 15 micros. The quantum yield of the signaling-state formation was determined to be 24%. Thus, the signaling state of the AppA BLUF domain is formed on the ultrafast time scale directly from the FAD singlet excited state, without any apparent intermediate, and remains stable over 12 decades of time. In parallel with the signaling state, the FAD triplet state is formed from the FAD singlet excited state at 9% efficiency as a side reaction of the AppA photocycle. PMID- 15751943 TI - Structural and spectral response of Aequorea victoria green fluorescent proteins to chromophore fluorination. AB - Global replacements of tyrosine by 2- and 3-fluorotyrosine in "enhanced green" and "enhanced yellow" mutants of Aequorea victoria green fluorescent proteins (avGFPs) provided protein variants with novel biophysical properties. While crystallographic and modeled structures of these proteins are indistinguishable from those of their native counterparts (i.e., they are perfectly isomorphous), there are considerable differences in their spectroscopic properties. The fluorine being an integral part of the avGFP chromophore induces changes in the titration curves, variations in the intensity of the absorbance and fluorescence, and spectral shifts in the emission maxima. Furthermore, targeted fluorination in close proximity to the fluorinated chromophore yielded additional variants with considerably enhanced spectral changes. These unique spectral properties are intrinsic features of the fluorinated avGFPs, in the context of the rigid chromophore-microenvironment interactions. The availability of the isomorpohous crystal structures of fluorinated avGFPs allowed mapping of novel, unusual interaction distances created by the presence of fluorine atoms. In addition, fluorine atoms in the ortho position of the chromophore tyrosyl moiety exhibit a single conformation, while in the meta position two conformer states were observed in the crystalline state. Such global replacements in chromophores of avGFPs and similar proteins result in "atomic mutations" (i.e., H --> F replacements) in the structures, offering unprecedented opportunities to understand and manipulate the relationships between protein structure and spectroscopic properties. PMID- 15751944 TI - Molecular basis for the origin of differential spectral and binding profiles of dansylamide with human carbonic anhydrase I and II. AB - Sulfonamide derivatives serve as potent inhibitors of carbonic anhydrases (CAs), and a few such inhibitors have been currently used as drugs for the treatment of different pathogenic conditions in humans. In pursuit of designing the isozyme specific inhibitors of human CAs, we observed that the fluorescence spectral properties and binding profiles of a fluorogenic sulfonamide derivative, 5 (dimethylamino)-1-naphthalenesulfonamide (dansylamide, DNSA), were markedly different between the recombinant forms of human carbonic anhydrase I (hCA I) and II (hCA II). The kinetic evaluation of the overall microscopic pathways for the binding of DNSA to hCA I versus hCA II revealed that the protein isomerization step served as a major determinant of the above discrepancy. Arguments are presented that the detailed structural-functional investigations of enzyme-ligand interactions may provide insights into designing the isozyme-specific inhibitors of CAs. PMID- 15751945 TI - Microstructure of cell wall-associated melanin in the human pathogenic fungus Cryptococcus neoformans. AB - Melanin is a virulence factor for many pathogenic fungal species, including Cryptococcus neoformans. Melanin is deposited in the cell wall, and melanin isolated from this fungus retains the shape of the cells, resulting in hollow spheres called "ghosts". In this study, atomic force, scanning electron, and transmission electron microscopy revealed that melanin ghosts are covered with roughly spherical granular particles approximately 40-130 nm in diameter, and that the melanin is arranged in multiple concentric layers. Nuclear magnetic resonance cryoporometry indicated melanin ghosts contain pores with diameters between 1 and 4 nm, in addition to a small number of pores with diameters near 30 nm. Binding of the antibodies to melanin reduced the apparent measured volume of these pores, suggesting a mechanism for their antifungal effect. We propose a model of cryptococcal melanin structure whereby the melanin granules are held together in layers. This structural model has implications for cell division, cell wall remodeling, and antifungal drug discovery. PMID- 15751946 TI - Conformational substates of calmodulin revealed by single-pair fluorescence resonance energy transfer: influence of solution conditions and oxidative modification. AB - A calmodulin (CaM) mutant (T34,110C-CaM) doubly labeled with fluorescence probes AlexaFluor 488 and Texas Red in opposing domains (CaM-DA) has been used to examine conformational heterogeneity in CaM by single-pair fluorescence resonance energy transfer (spFRET). Burst-integrated FRET efficiencies of freely diffusing CaM-DA single molecules yielded distributions of distance between domains of CaM DA. We recently reported distinct conformational substates of Ca(2+)-CaM-DA and apoCaM-DA, with peaks in the distance distributions centered at approximately 28 A, 34-38 A, and 55 A [Slaughter et al. (2004) J. Phys. Chem. B 108, 10388-10397]. In the present study, shifts in the amplitudes and center distances of the conformational substates were detected with variation in solution conditions. The amplitude of an extended conformation was observed to change as a function of Ca(2+) over a free Ca(2+) range that is consistent with binding to the high affinity, C-terminal Ca(2+) binding sites, suggesting the existence of communication between lobes of CaM. Lowering pH shifted the relative amplitudes of the conformations, with a marked increase in the presence of the compact conformations and an almost complete absence of the extended conformation. In addition, the single-molecule distance distribution of apoCaM-DA at reduced ionic strength was shifted to longer distance and showed evidence of an increase in conformational heterogeneity relative to apoCaM-DA at physiological ionic strength. Oxidation of methionine residues in CaM-DA produced a substantial increase in the amplitude of the extended conformation relative to the more compact conformation. The results are considered in light of a hypothesis that suggests that electrostatic interactions between charged amino acid side chains play an important role in determining the most stable CaM conformation under varying solution conditions. PMID- 15751947 TI - NMR structural study of TcUBP1, a single RRM domain protein from Trypanosoma cruzi: contribution of a beta hairpin to RNA binding. AB - TcUBP1 is a trypanosome cytoplasmic RNA-binding protein containing a single, conserved RNA-recognition motif (RRM) domain involved in selective destabilization of U-rich mRNAs such as the Trypanosoma cruzi small mucin gene family mRNA, TcSMUG. TcUBP1 binds specific transcripts in vivo and co-localizes in the perinuclear part of the cell with components of the mRNA-stability determinant pathway such as poly(A)-binding protein 1 (PABP1) and TcUBP2, a closely related RRM-containing protein. In TcUBP proteins, the RRM domain is flanked by N-terminal Gln-rich and C-terminal Gly-Gln-rich extensions, which are involved in protein-protein interactions. In this work, we determined the solution structure of the TcUBP1 RRM domain by nuclear magnetic resonance (NMR) spectroscopy. The domain has a characteristic betaalphabetabetaalphabeta fold, consisting of a beta sheet composed of four antiparallel betastrands and two alpha helices packed against one face of the beta sheet. A unique aspect of TcUBP1 is the participation of a beta hairpin (beta4-beta5) in the beta sheet, resulting in an enlarged RNA-binding surface. Detailed analysis of the TcUBP1 interaction with a short single-stranded RNA derived from the 3' UTR of TcSMUG was carried out by titration experiments using both NMR spectroscopy and isothermal titration calorimetry. This analysis revealed that amino acids located within the beta hairpin (beta4-beta5) contribute to complex formation. This enlarged protein-RNA interface could compensate for the lack of additional RNA binding domains in TcUBP1, as observed in many other RRM-containing proteins. The structure of TcUBP1 reveals new aspects of single RRM-RNA interactions and insight into how N- and C-terminal extensions can contribute to RNA binding. PMID- 15751948 TI - Conjugated polyelectrolytes: conformation-sensitive optical probes for detection of amyloid fibril formation. AB - The in vivo deposition of amyloid fibrils is a hallmark of many devastating diseases known as the amyloidoses. Amyloid formation in vitro may also complicate production of proteins in the biotechnology industry. Simple, sensitive, and versatile tools that detect the fibrillar conformation of amyloidogenic proteins are thus of great importance. We have developed a negatively charged conjugated polyelectrolyte that displays different characteristic optical changes, detected visually or by absorption and emission, depending on whether the protein with which it forms a complex is in its native state or amyloid fibril conformation. This simple, rapid, and novel methodology was applied here to two amyloidogenic proteins, insulin and lysozyme, and its validity for detection of their fibrillar conformation was verified by currently used methods such as circular dichroism, transmission electron microscopy, and Congo red absorption. PMID- 15751949 TI - Illustration of HIV-1 protease folding through a molten-globule-like intermediate using an experimental model that implicates alpha-crystallin and calcium ions. AB - The folding of HIV-1 protease to its active form involves the coordination of structure formation and dimerization, which follows a hierarchy consisting of folding nuclei spanning from the active site, hinge region, and dimerization domain. However, the biochemical characteristics of the folding intermediates of this protein remain to be elucidated. In an experimental model, the denaturation of the tethered dimer of HIV-1 protease by guanidine hydrochloride revealed an alternative conformation resembling the molten-globule state. The molten-globule state binds to the molecular chaperone alpha-crystallin and prevents its aggregation; however, the chaperone alone failed to reconstitute HIV-1 protease into its active form. Calcium ion assisted in the release of active enzyme from the chaperone complex. Alpha-crystallin, a member of the small heat-shock protein, assists proteins to fold correctly; however, the underlying principle of signals responsible for chaperone-mediated protein folding remains enigmatic. X ray photoelectron spectroscopy has been employed to provide the evidence of calcium binding to alpha-crystallin and to decipher the effect of calcium binding on the chaperone-mediated refolding of HIV-1 protease. On the basis of our spectroscopic data, we propose that calcium ions interact with the carboxyl groups of the surface-exposed acidic amino acids of alpha-crystallin bringing electrostatic interference, which plays a pivotal role in inducing conformational changes in the chaperone responsible for the release of the active enzyme. PMID- 15751950 TI - Composition and sequence-dependent binding of RNA to the nucleocapsid protein of Moloney murine leukemia virus. AB - All retroviruses package two copies of their genomes during virus assembly, both of which are required for strand transfer-mediated recombination during reverse transcription. Genome packaging is mediated by interactions between the nucleocapsid (NC) domains of assembling Gag polyproteins and an RNA packaging signal, located near the 5' end of the genome, called Psi. We recently discovered that the NC protein of the Moloney murine leukemia virus (MLV) can bind with high affinity to conserved UCUG elements within the MLV packaging signal [D'Souza, V., and Summers, M. F. (2004) Nature 431, 586-590]. Selective binding to dimeric RNA is regulated by a conformational RNA switch, in which the UCUG elements are sequestered by base pairing in the monomeric RNA and do not bind NC, but become exposed for NC binding upon dimerization. Dimerization-dependent structural changes occur in other regions of the Psi-site, exposing guanosine-containing segments that might also bind NC. Here we demonstrate that short RNAs containing three such sequences, ACAG, UUUG, and UCCG, can bind NC with significant affinity (K(d) = 94-315 nM). Titration experiments with oligoribonucleotides of varying lengths and compositions, combined with NMR-based structural studies, reveal that binding is strictly dependent on the presence of an unpaired guanosine, and that relative binding affinities can vary by more than 1 order of magnitude depending on the nature of the three upstream nucleotides. Binding is enhanced in short RNAs containing terminal phosphates, indicating that electrostatic interactions contribute significantly to binding. Our findings extend a previously published model for genome recognition, in which the NC domains of assembling Gag molecules interact with multiple X(i-3)-X(i-2)-X(i-1)-G(i) elements (X is a variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. PMID- 15751951 TI - Crystal structure of S-ribosylhomocysteinase (LuxS) in complex with a catalytic 2 ketone intermediate. AB - S-Ribosylhomocysteinase (LuxS) is an Fe(2+)-dependent metalloenzyme that catalyzes the cleavage of the thioether bond in S-ribosylhomocysteine (SRH) to produce homocysteine (Hcys) and 4,5-dihydroxy-2,3-pentanedione (DPD), the precursor of type II bacterial quorum-sensing molecule. The proposed mechanism involves an initial metal-catalyzed aldose-ketose isomerization reaction, which results in the migration of the ribose carbonyl group from its C1 to C2 position and the formation of a 2-ketone intermediate. A repetition of the isomerization reaction shifts the carbonyl group to the C3 position. Subsequent beta elimination reaction at the C4 and C5 positions completes the catalytic cycle. In this work, a catalytically inactive mutant (C84A) of Co(2+)-substituted Bacillus subtilis LuxS was cocrystallized with the 2-ketone intermediate and the structure was determined to 1.8 A resolution. The structure reveals that the C2 carbonyl oxygen is directly coordinated with the metal ion, providing strong support for the proposed Lewis acid function of the metal ion during catalysis. Cys-84 and Glu-57 are optimally positioned to act as general acids/bases during the isomerization and elimination reactions. In addition, Ser-6, His-11, and Arg-39 are involved in substrate/ intermediate binding through hydrogen bonding interactions. The above conclusions are further confirmed by site-directed mutagenesis and visible absorption spectroscopic studies. PMID- 15751952 TI - Experimental demonstration of T:(G:G:G:G):T hexad and T:A:A:T tetrad alignments within a DNA quadruplex stem. AB - A template-based approach was used to design unprecedented architectural motifs into a known DNA framework. The structure formed by the sequence d(GCGGTTGGAT) in 0.1 M Na(+) solution has been determined using molecular dynamics simulations constrained by distance and dihedral restraints derived from NMR experiments. The molecular topology has been previously observed for the sequence d(GCGGTGGAT) (Webba da Silva, M. (2003) Biochemistry 42, 14356-65). Insertion of a single thymine into the double chain reversal formed by the segment GGTGG results in the unprecedented experimental demonstration of a T:(G:G:G:G):T hexad. The bi stranded hexad results from the pairing alignment of two G(T-G) triads. Each triad results from recognition of the sheared edge of a guanine by the Watson Crick edge of a thymine of the segment GGTTGG. The alignment is stabilized by base-stacking of the thymine to the sugar pucker of the preceding thymine. The latter is involved in formation of the T:A:A:T tetrad alignment by forming a hydrogen bond with the free amino proton of a Watson-Crick aligned A:A mispair. We have thus established that residues in double chain reversal loops linking juxtaposed tetrads of a quadruplex stem may facilitate formation of yet unknown hydrogen bond alignments. By employing a systematic approach analysis of sequence motifs appearing in double chain reversals, bridging tetrad layers should allow for the prediction of topologies and architectural motifs appearing in biologically relevant genomic regions. PMID- 15751953 TI - Secondary-structure characterization of two proficient kinase deoxyribozymes. AB - Dk1 and Dk2 are two catalytically proficient, manganese-dependent, guanine-rich deoxyribozymes previously isolated for DNA phosphorylation. In this study, we carried out a series of experiments that aimed to understand the structural properties of Dk1 and Dk2 and compare the structural similarities or differences of these two distinct deoxyribozymes that carry out similar catalytic functions. First, we performed reselections from two partially randomized DNA libraries on the basis of the original Dk1 and Dk2 sequences to isolate catalytically active sequence variants and identify nucleotides that are invariable, well-conserved, or highly mutagenized. Sequence analysis of these variants assisted by secondary structure predictions led to the identification of possible Watson-Crick base pairing regions within each deoxyribozyme. Sequence truncation and base-pair partner exchange experiments were conducted to confirm, or rule out, the existence of the predicted secondary-structure elements. Finally, methylation interference experiments were applied to identify nucleotides that are potentially important for the tertiary structure folding of the deoxyribozymes. Our data suggest that Dk1 and Dk2, despite the differences in their primary sequences and NTP requirements, use an analogous stem-loop element to anchor a structural domain of substantial tertiary interactions to execute their catalytic functions. PMID- 15751954 TI - Hinge residue Ile260 of DNA polymerase beta is important for enzyme activity and fidelity. AB - DNA polymerases ensure efficient insertion of the correct dNTP into the DNA substrate. They have evolved mechanisms for discriminating among very similar dNTP substrates. DNA polymerase beta is a repair polymerase that provides a model system for a direct study of insertion fidelity. In this study, we examined the role of hinge residue Ile260 of the rat Polbeta on enzyme activity and accuracy. We changed residue I260 to every other amino acid residue and used genetic screens to assess the activity and fidelity of the resulting mutants. The I260D, E, -K, -N, and -R mutants are significantly less active than wild-type Polbeta. Interestingly, I260H and I260Q are active but exhibit mutator activity. This suggests that the nonpolar nature of residue 260 is important for maintaining the activity and fidelity of Polbeta. We employ molecular modeling as an aid in explaining the observed phenotypes and propose a mechanism whereby the positioning of the DNA substrate in the enzyme and within the surface of the hinge may be a key player in forming an optimal active site for phosphodiester bond formation between Watson-Crick base pairs. PMID- 15751955 TI - A substrate-assisted concerted mechanism for aminoacylation by a class II aminoacyl-tRNA synthetase. AB - Aminoacyl-tRNA synthetases (aaRS) join amino acids to their cognate transfer RNAs, establishing an essential coding relationship in translation. To investigate the mechanism of aminoacyl transfer in class II Escherichia coli histidyl-tRNA synthetase (HisRS), we devised a rapid quench assay. Under single turnover conditions with limiting tRNA, aminoacyl transfer proceeds at 18.8 s( )(1), whereas in the steady state, the overall rate of aminoacylation is limited by amino acid activation to a rate of 3 s(-)(1). In vivo, this mechanism may serve to allow the size of amino acid pools and energy charge to control the rate of aminoacylation and thus protein synthesis. Aminoacyl transfer experiments using HisRS active site mutants and phosphorothioate-substituted adenylate showed that substitution of the nonbridging Sp oxygen of the adenylate decreased the transfer rate at least 10 000-fold, providing direct experimental evidence for the role of this group as a general base for the reaction. Other kinetic experiments revealed that the rate of aminoacyl transfer is independent of the interaction between the carboxyamide group of Gln127 and the alpha-carboxylate carbon, arguing against the formation of a tetrahedral intermediate during the aminoacyl transfer. These experiments support a substrate-assisted concerted mechanism for HisRS, a feature that may generalize to other aaRS, as well as the peptidyl transferase center. PMID- 15751956 TI - Role of structural plasticity in signal transduction by the cryptochrome blue light photoreceptor. AB - Cryptochromes are blue-light photoreceptors that regulate a variety of responses such as growth and circadian rhythms in organisms ranging from bacteria to humans. Cryptochromes share a high level of sequence identity with the light activated DNA repair enzyme photolyase. Photolyase uses energy from blue light to repair UV-induced photoproducts in DNA through cyclic electron transfer between the catalytic flavin adenine dinucleotide cofactor and the damaged DNA. Cryptochromes lack DNA repair activity, and their mechanism of signal transduction is not known. It is hypothesized that a light-dependent signaling state in cryptochromes is created as a result of an intramolecular redox reaction, resulting in conformational rearrangement and effector binding. Plant and animal cryptochromes possess 30-250 amino acid carboxy-terminal extensions beyond the photolyase-homology region that have been shown to mediate phototransduction. We analyzed the structures of C-terminal domains from an animal and a plant cryptochrome by computational, biophysical, and biochemical methods and found these domains to be intrinsically unstructured. We show that the photolyase-homology region interacts with the C-terminal domain, inducing stable tertiary structure in the C-terminal domain. Importantly, we demonstrate a light-dependent conformational change in the C-terminal domain of Arabidopsis Cry1. Collectively, these findings provide the first biochemical evidence for the proposed conformational rearrangement of cryptochromes upon light exposure. PMID- 15751957 TI - Intersubunit interactions associated with Tyr42 alpha stabilize the quaternary-T tetramer but are not major quaternary constraints in deoxyhemoglobin. AB - Previous mutational studies on Tyr42alpha variants as well as the current studies on the mutant hemoglobin alphaY42A show that the intersubunit interactions associated with Tyr42alpha significantly stabilize the alpha1beta2 interface of the quaternary-T deoxyhemoglobin tetramer. However, crystallographic studies, UV and visible resonance Raman spectroscopy, CO combination kinetic measurements, and oxygen binding measurements on alphaY42A show that the intersubunit interactions formed by Tyr42alpha have only a modest influence on the structural properties and ligand affinity of the deoxyhemoglobin tetramer. Therefore, the alpha1beta2 interface interactions associated with Tyr42alpha do not contribute significantly to the quaternary constraints that are responsible for the low oxygen affinity of deoxyhemoglobin. The slight increase in the ligand affinity of deoxy alphaY42A correlates with small, mutation-induced structural changes that perturb the environment of Trp37beta, a critical region of the quaternary-T alpha1beta2 interface that has been shown to be the major source of quaternary constraint in deoxyhemoglobin. PMID- 15751958 TI - Phosphorylation of bovine adrenodoxin by protein kinase CK2 affects the interaction with its redox partner cytochrome P450scc (CYP11A1). AB - Adrenodoxin (Adx), a [2Fe-2S] vertebrate-type ferredoxin, transfers electrons from the NADPH-dependent flavoprotein Adx reductase (AdR) to mitochondrial cytochrome P450 enzymes of the CYP11A and CYP11B families, which catalyze key reactions in steroid hormone biosynthesis. Adx is a known phosphoprotein, but the kinases that phosphorylate Adx have remained mostly obscure. The aim of this study was to identify previously unknown Adx phosphorylating kinases and to acquire a deeper insight into the functional consequences of such a modification. Here, we show for the first time that bovine Adx is a substrate of protein kinase CK2, whereas bovine CYP11A1, CYP11B1, and AdR are not phosphorylated by this kinase. CK2 phosphorylation of mature Adx requires the presence of both the catalytic alpha-subunit and the regulatory beta-subunit of CK2 and takes place exclusively at residue Thr-71, which is located within the redox partner interaction domain of the protein. We created two Adx mutants, Adx-T71E (imitating a phosphorylation) and Adx-T71V (which cannot be phosphorylated at this site), respectively, and investigated how these mutations affected the interaction of Adx with its redox partners. These data were supplemented with detailed spectroscopic and functional assays using the phosphorylated protein. All Adx species behaved like wild type (Adx-WT) with respect to their redox potential, iron-sulfur cluster symmetry, and overall backbone structure. Substrate conversion assays catalyzed by CYP11A1 showed an increase in product formation when Adx-T71E or CK2-phosphorylated Adx were used as electron carrier instead of Adx-WT, whereas the activity toward CYP11B1 was not altered using these Adx species. Additionally, Adx-T71E represents the only full-length Adx mutant which leads to an increase in CYP11A1 product formation. Therefore, characterizing this full-length mutant helps to improve our knowledge on the functional effects of phosphorylations on complex redox systems. PMID- 15751959 TI - P450 active site architecture and reversibility: inactivation of cytochromes P450 2B4 and 2B4 T302A by tert-butyl acetylenes. AB - The inactivations of P450 2B4 and the T302A mutant of 2B4 by tert-butyl acetylene (tBA) and the inactivation of 2B4 T302A by tert-butyl 1-methyl-2-propynyl ether (tBMP) have been investigated. tBA and tBMP inactivated both enzymes in a mechanism-based manner with the losses in enzymatic activity corresponding closely to losses in P450 heme. HPLC and ESI-LC-MS analysis detected two different tBA- or tBMP-modified heme products with masses of 661 and 705 Da, respectively. Interestingly, the inactivations of the P450s 2B4 by tBA and tBMP were partially reversible by dialysis, and the tBA- or tBMP-modified heme products could only be observed with ESI-LC-MS/MS when the inactivated samples were acidified prior to analysis, indicating a requirement for protons in the formation of stable heme adducts in both the wild-type and mutant 2B4 enzymes. Results of studies using artificial oxidants to support enzyme inactivation suggest that the oxenoid-iron activated oxygen species is preferentially utilized during the inactivation of the P450s 2B4 by tBA. These results argue against the use of a peroxo-iron species by P450 2B4 T302A. Molecular dynamics studies of wild-type P450 2B4 reveal that contiguous hydrogen bond networks, including structural waters, link a conserved glutamate (E301) to the distal oxygen of the peroxo-heme species via threonine 302. Interestingly, models of 2B4 T302A reveal that a compensatory, ordered hydrogen bond network forms despite the removal of T302. These results indicate that while T302 may play a role in proton delivery in the formation of the oxenoid-iron complex and in the stabilization of acetylene heme adducts in 2B4, it is not essential for proton delivery given the presence of E301 in the binding site. PMID- 15751960 TI - Steady-state and transient kinetic analyses of taurine/alpha-ketoglutarate dioxygenase: effects of oxygen concentration, alternative sulfonates, and active site variants on the FeIV-oxo intermediate. AB - Taurine/alpha-ketoglutarate (alphaKG) dioxygenase (TauD), an archetype alphaKG dependent hydroxylase, is a non-heme mononuclear Fe(II) enzyme that couples the oxidative decarboxylation of alphaKG with the conversion of taurine to aminoacetaldehyde and sulfite. The crystal structure of taurine-alphaKG Fe(II)TauD is known, and spectroscopic studies have kinetically defined the early steps in catalysis and identified a high-spin Fe(IV)-oxo reaction intermediate. The present analysis extends our understanding of TauD catalysis by investigating the steady-state and transient kinetics of wild-type and variant forms of the enzyme with taurine and alternative sulfonates. TauD proteins substituted at residues surrounding the active site were shown to fold properly based on their abilities to form a diagnostic chromophore associated with the anaerobic Fe(II) alphaKG chelate complex and to generate a tyrosyl radical upon subsequent reaction with oxygen. Steady-state studies of mutant proteins confirmed the importance of His 70 and Arg 270 in binding the sulfonate moiety of taurine and indicated the participation of Asn 95 in recognizing the substrate amine group. The N97A and S158A variants are likely to undergo an increase in hydrophobicity and expansion of the substrate-binding pocket, thus accounting for their decreased K(m) toward pentanesulfonic acid compared to wild-type TauD. Stopped flow UV-visible spectroscopic examination of the reaction of oxygen with taurine alphaKG-Fe(II)TauD confirmed a minimal three-step sequence of reactions attributed to Fe(IV)-oxo formation (k(1)), bleaching to the Fe(II) state upon substrate hydroxylation (k(2)), rebinding of excess substrates (k(3)), and indicated that none of the steps exhibit detectable solvent k(H)/k(D) isotope effects. This demonstrates that no protons are involved in the rate-determining step of Fe(IV)-oxo formation, in contrast to heme iron oxygenases. The Fe(IV)-oxo species is likely to be utilized in conversion of the alternative substrates pentanesulfonic acid and 3-N-morpholinopropanesulfonic acid; however, this spectroscopic intermediate was not detected because of the decreased k(1)/k(2) ratio. With taurine, k(1) was shown to depend on the oxygen concentration allowing calculation of a second-order rate constant of 1.58 x 10(5) M(-)(1) s( )(1) for this irreversible reaction. Stopped-flow analyses of TauD variants provided several insights into how the protein environment influences the rates of Fe(IV)-oxo formation and decay. The Fe(IV)-oxo species was not detected in the N95D or N95A variants because of a reduced k(1)/k(2) ratio, likely related to a decreased substrate-dependent conversion of the six-coordinate to five-coordinate metal site. PMID- 15751961 TI - Distance and affinity dependence of triplex-induced recombination. AB - Triplex-forming oligonucleotides (TFOs) have the potential to serve as gene therapeutic agents on the basis of their ability to mediate site-specific genome modification via induced recombination. However, high-affinity triplex formation is limited to polypurine/polypyrimidine sites in duplex DNA. Because of this sequence restriction, careful analysis is needed to identify suitable TFO target sites within or near genes of interest. We report here an examination of two key parameters which influence the efficiency of TFO-induced recombination: (1) binding affinity of the TFO for the target site and (2) the distance between the target site and the mutation to be corrected. To test the influence of binding affinity, we compared induced recombination in human cell-free extracts by a series of G-rich oligonucleotides with an identical base composition and an increasing number of mismatches in the third strand binding code. As the number of mismatches increased and, therefore, binding affinity decreased, induced recombination frequency also dropped. There was an apparent threshold at an equilibrium dissociation constant (K(d)) of 1 x 10(-)(7) M. In addition, TFO chemical modification with N,N-diethylethylenediamine (DEED) internucleoside linkages to confer improved binding was found to yield increased levels of induced recombination. To test the ability of triplex formation to induce recombination at a distance, episomal targets with informative reporter genes were constructed to contain polypurine TFO target sites at varying distances from the mutations to be corrected. TFO-induced recombination in mammalian cells between a plasmid vector and a donor oligonucleotide was detected at distances ranging from 24 to 750 bp. Together, these results indicate that TFO-induced recombination requires high-affinity binding but can affect sites hundreds of base pairs away from the position of triplex formation. PMID- 15751962 TI - Binding of six nucleotide cofactors to the hexameric helicase RepA protein of plasmid RSF1010. 1. Direct evidence of cooperative interactions between the nucleotide-binding sites of a hexameric helicase. AB - The interactions of nucleotides with RepA hexameric helicase from plasmid RSF1010 have been examined using nucleotide analogues, TNP-ADP, TNP-ATP, and MANT-ADP. The binding of the analogues is accompanied by strong quenching of the protein fluorescence. A quantitative fluorescence titration method has been applied to analyze the interactions, independent of any assumptions of proportionality between the fluorescence quenching and the average degree of binding. The fluorescence quenching as a function of the average degree of binding is expressed by an empirical function that enables analysis of the data, without the necessity of determining quenching parameters for different complexes. At saturation, the RepA hexamer binds six nucleotide molecules, indicating that each subunit of the hexamer can engage in interactions with the cofactor. The nucleotide macroscopic affinity decreases with the increasing degree of binding, indicating heterogeneity among the binding sites. A statistical thermodynamic hexagon model provides an excellent description of the binding process and requires only two interaction parameters, the intrinsic binding constant, K, and cooperativity parameter, sigma. The heterogeneity in affinity reflects negative cooperative interactions between the binding sites. Analyses of the data provide clear evidence that the alternative model of two independent classes of binding sites does not describe the nucleotide binding. Such a model cannot account for both, the binding isotherms and the dependence of the fluorescence quenching upon the degree of binding. Thus, cooperative interactions between the nucleotide binding sites are an intrinsic property of the RepA helicase. The presence of the cooperative interactions indicates significant communication among the subunits of the helicase. PMID- 15751963 TI - Binding of six nucleotide cofactors to the hexameric helicase RepA protein of plasmid RSF1010. 2. Base specificity, nucleotide structure, magnesium, and salt effect on the cooperative binding of the cofactors. AB - Interactions of the RepA hexameric helicase with nucleotide cofactors have been examined using nucleotide analogues, TNP-ADP and TNP-ATP, and unmodified nucleotides. Thermodynamic parameters for the interactions of modified and unmodified nucleotides have been obtained using quantitative fluorescence titration and competition titration methods. The intrinsic binding constant of ATP is by a factor of approximately 10 and approximately 1000 higher than the value observed for ADP and PO(4)(-). The data suggest that helicase acquires free energy transducing capabilities when associated with the ssDNA, thus, forming a "holoenzyme". ATP binding is characterized by significantly stronger negative cooperativity than ADP. The cooperative interactions are predominantly induced through the specific interactions of the gamma phosphate and the ribose with the protein. The salt effect on cofactor binding indicates a very different nature of the intrinsic and cooperative interactions. Surprisingly, binding of Mg(2+), to both the cofactor and helicase, predominantly controls the ADP-RepA interactions. Mg(2+) cations seem to play a role in affecting the distribution of high and low ssDNA-affinity states, through the strong effect on the diphosphate versus triphosphate binding. The data indicate that Mg(2+) has a dual function in nucleotide-helicase interactions. At low [Mg(2+)], NTP binds stronger than NDP and the enzyme is predominantly in the high ssDNA-affinity state. At higher [Mg(2+)], NTP binds weaker than NDP and the helicase subunits can exist in alternating low- and high-affinity states that facilitate the efficient dsDNA unwinding. The RepA helicase shows a preference toward purine nucleotides. The cooperative interactions are independent of the type of the base. PMID- 15751965 TI - Chlorella virus Marburg topoisomerase II: high DNA cleavage activity as a characteristic of Chlorella virus type II enzymes. AB - Although the formation of a covalent enzyme-cleaved DNA complex is a prerequisite for the essential functions of topoisomerase II, this reaction intermediate has the potential to destabilize the genome. Consequently, all known eukaryotic type II enzymes maintain this complex at a low steady-state level. Recently, however, a novel topoisomerase II was discovered in Paramecium bursaria chlorella virus-1 (PBCV-1) that has an exceptionally high DNA cleavage activity [Fortune et al. (2001) J. Biol. Chem. 276, 24401-24408]. If robust DNA cleavage is critical to the physiological functions of chlorella virus topoisomerase II, then this remarkable characteristic should be conserved throughout the viral family. Therefore, topoisomerase II from Chlorella virus Marburg-1 (CVM-1), a distant family member, was expressed in yeast, isolated, and characterized. CVM-1 topoisomerase II is 1058 amino acids in length, making it the smallest known type II enzyme. The viral topoisomerase II displayed a high DNA strand passage activity and a DNA cleavage activity that was approximately 50-fold greater than that of human topoisomerase IIalpha. High DNA cleavage appeared to result from a greater rate of scission rather than promiscuous DNA site utilization, inordinately tight DNA binding, or diminished religation rates. Despite the fact that CVM-1 and PBCV-1 topoisomerase II share approximately 67% amino acid sequence identity, the two enzymes displayed clear differences in their DNA cleavage specificity/site utilization. These findings suggest that robust DNA cleavage is intrinsic to the viral enzyme and imply that chlorella virus topoisomerase II plays a physiological role beyond the control of DNA topology. PMID- 15751964 TI - Tyrosine phosphorylation of adaptor protein 3BP2 induces T cell receptor-mediated activation of transcription factor. AB - Molecular adaptors/scaffolds have indispensable roles in the activation of lymphocytes. In this report, we have demonstrated the role of tyrosine phosphorylation of an adaptor protein 3BP2 (c-Abl-SH3 domain binding protein-2, also known as SH3BP2) in T cell receptor (TCR)-mediated activation of transcription factor. Short interfering RNA for 3BP2 suppresses the expression level of endogenous 3BP2 and inhibits TCR-mediated activation of interleukin (IL) 2 promoter and nuclear factor of activated T cells (NFAT) element. Engagement of TCR induces tyrosine phosphorylation and lipid raft translocation of 3BP2. The overexpression studies reveal that substitution of 3BP2-Tyr(183), Tyr(446), or Arg(486) in the SH2 domain suppresses TCR-mediated activation of NFAT. Point mutations of 3BP2 cannot affect the translocation of 3BP2 into the lipid raft. Phosphorylation of Tyr(183) is required for the interaction with Vav1, the guanine nucleotide exchanging factor of Rac1. In fact, overexpression of dominant negative form of Rac1 inhibits TCR-mediated activation of NFAT. Phosphorylation of Tyr(446) recruits the SH2 domain of Lck for the optimal activation of transcription factors. Furthermore, point mutation of Arg(486) in the 3BP2-SH2 domain that couples ZAP-70 to LAT dramatically reduces NFAT activation. These results suggest that the site-directed functions of 3BP2 induce the activation of transcription factors. PMID- 15751966 TI - Induction of a cell stress response gene RTP801 by DNA damaging agent methyl methanesulfonate through CCAAT/enhancer binding protein. AB - RTP801 is a newly discovered stress response gene that is induced by hypoxia and other cell stress signals. Here, we investigated the mechanism by which a DNA damaging agent, methyl methanesulfonate (MMS), induces RTP801 transcription. In HaCaT human keratinocytes, MMS was able to induce a rapid increase in the mRNA level of RTP801. Correspondingly, MMS treatment was capable of stimulating a 2.5 kb RTP801 promoter. Deletion studies with the promoter demonstrated a critical region between -1057 and -981 bp of the promoter that is responsive to MMS treatment. Point mutations of the consensus Elk-1 and C/EBP sites within this region were able to abrogate the stimulatory effect of MMS, indicating that Elk-1 and C/EBP are both involved in the transcriptional regulation of the RTP801 gene by MMS. Furthermore, a gel mobility shift assay revealed that MMS was able to initiate rapid formation of a protein complex that bound the C/EBP site of the promoter. In addition, an anti-C/EBPbetaantibody was capable of further shifting the bound protein complex. Therefore, these studies indicate that RTP801 is a transcriptional target of MMS in human keratinocytes and that C/EBP is implicated in transcriptional control of the gene. PMID- 15751967 TI - A complementary single-stranded docking site is required for enhancement of strand exchange by human immunodeficiency virus nucleocapsid protein on substrates that model viral recombination. AB - Enhancement of strand exchange by nucleocapsid protein (NC) is proposed to occur during retroviral recombination. The mechanism was examined using an RNA (donor) DNA hybrid that mimicked a retrovirus replication intermediate. This consisted of a 25 base pair hybrid region flanked on each side by single-stranded RNA or DNA. A second set of acceptor RNAs that could bind to the 25-base hybrid region and to various lengths of additional bases on the DNA was used to displace the donor by hybridizing with the DNA. Displacement required a complementary single-stranded DNA region outside the donor-DNA 25-nucleotide hybrid region. NC enhanced displacement slightly when the acceptor could bind 10 nucleotides and significantly when binding 22 or more nucleotides in the single-stranded region. Two mutated acceptors that bound over 47 total nucleotides on the DNA (22 in the single-stranded region plus 25 in the hybrid region) were constructed. One had three mismatches in the hybrid region; the other, three in the single-stranded region and one in the hybrid region. Each acceptor bound the DNA with approximately equal thermodynamic stability, yet NC stimulated exchange with the former and actually inhibited with the latter. This emphasized the importance of the single-stranded region in NC stimulation. The results support a mechanism where NC enhances the docking of the acceptor to the single-stranded region and then the acceptor "zippers" through the hybrid and displaces the donor. Results with the mutated acceptors indicate that NC may actually inhibit strand exchange between genomes in nonhomologous regions. PMID- 15751968 TI - Ezrin mutants affecting dimerization and activation. AB - ERM (ezrin/radixin/moesin) proteins provide a regulated linkage between membrane associated proteins and the actin cytoskeleton. Previous work has shown that ezrin can exist in a dormant monomeric state in which the N-terminal FERM domain is tightly associated with the C-ERMAD (carboxyl-terminal ERM association domain), masking binding sites for at least some ligands, including F-actin and the scaffolding protein EBP50. Activation of ezrin requires relief of the intramolecular association, and this is believed to involve phosphorylation of threonine 567. Studies have therefore employed the T567D phosphomimetic mutant to explore the consequences of ezrin activation in vivo. Ezrin also exists as a stable dimer, in which the orientation of the two subunits is unknown, but might involve the central alpha-helical region predicted to form a coiled-coil. By characterization of ezrin mutants, we show that relief of the intramolecular association in the monomer results in unmasking of ligand binding sites and a significant conformational change, that the T567D mutation has a small effect on the biochemical activation of ezrin, and that the predicted coiled-coil region does not drive dimer formation. These results provide strong support for the conformational activation model of ezrin, elucidate the basis for dimer formation, and reveal that a mutant generally considered to be fully activated is not. PMID- 15751969 TI - Defined sites of interaction between subunits E (Vma4p), C (Vma5p), and G (Vma10p) within the stator structure of the vacuolar H+-ATPase. AB - Vacuolar H(+)-ATPases (V-ATPases) are multi-subunit membrane proteins that couple ATP hydrolysis to the extrusion of protons from the cytoplasm. Although they share a common macromolecular architecture and rotational mechanism with the F(1)F(0)-ATPases, the organization of many of the specialized V-ATPase subunits within this rotary molecular motor remains uncertain. In this study, we have identified sequence segments involved in linking putative stator subunits in the Saccharomyces V-ATPase. Precipitation assays revealed that subunits Vma5p (subunit C) and Vma10p (subunit G), expressed as glutathione-S-transferase fusion proteins in E. coli, are both able to interact strongly with Vma4p (subunit E) expressed in a cell-free system. GST-Vma10p also associated with Vma2p and Vma1p, the core subunits of the ATP-hydrolyzing domain, and was able to self-associate to form a dimer. Mutations within the first 19-residue region of Vma4p, which disrupted interaction with Vma5p in vitro, also prevented the Vma4p polypeptide from restoring V-ATPase function in a complementation assay in vivo. These mutations did not prevent assembly of Vma5p (subunit C) and Vma2p (subunit B) into an inactive complex at the vacuolar membrane, indicating that Vma5p must make multiple interactions involving other V-ATPase subunits. A second, highly conserved region of Vma4p between residues 19 and 38 is involved in binding Vma10p. This region is highly enriched in charged residues, suggesting a role for electrostatic effects in Vma4p-Vma10p interaction. These protein interaction studies show that the N-terminal region of Vma4p is a key factor not only in the stator structure of the V-ATPase rotary molecular motor, but also in mediating interactions with putative regulatory subunits. PMID- 15751970 TI - Protein kinase C phosphorylation modulates N- and C-terminal regulatory activities of the PITX2 homeodomain protein. AB - PKC phosphorylation regulates PITX2 DNA binding and transcriptional activity. Mutation of individual PKC sites demonstrates the functional regulation of PITX2 through phosphorylation. Immunoprecipitation of PITX2 and a PITX2 PKC mutant protein reveal specific in vivo phosphorylation by PKC in transfected cells. The transcriptional activity of PITX2 is negatively regulated by N-terminal phosphorylation and positively regulated by C-terminal phosphorylation. We demonstrate a mechanism of increased PITX2 transcriptional activation through protein interactions facilitated by phosphorylation of the PITX2 C-terminal tail. Phosphorylation of the PITX2 C terminus enhances the interaction with cellular factors. In corroboration with the PITX2 PKC functional studies, a newly identified C-terminal PITX2 mutation associated with Axenfeld-Rieger syndrome (ARS) demonstrates reduced phosphorylation. This mutation (PITX2 DeltaT1261) creates a frameshift mutation in codon 227 resulting in 11 novel amino acids downstream followed by premature truncation of the protein. Three PKC sites in the C-terminal tail and OAR domain are deleted, which results in decreased transcriptional activation. PITX2 DeltaT1261 is unable to interact with a cellular factor to synergistically activate transcription and demonstrates the first link of ARS with defective PITX2 protein interactions. Gene expression profiling of homozygous Pitx2 mutant mouse tissue reveals decreased Dlx2 expression as a potential molecular basis for developmental defects associated with ARS patients. Overall, phosphorylation imparts another level of regulation to the activity of the PITX2 homeodomain protein during development. PMID- 15751971 TI - Primary contact sites in intrinsically unstructured proteins: the case of calpastatin and microtubule-associated protein 2. AB - Intrinsically unstructured proteins (IUPs) exist in a disordered conformational state, often considered to be equivalent with the random-coil structure. We challenge this simplifying view by limited proteolysis, circular dichroism (CD) spectroscopy, and solid-state (1)H NMR, to show short- and long-range structural organization in two IUPs, the first inhibitory domain of calpastatin (CSD1) and microtubule-associated protein 2c (MAP2c). Proteases of either narrow (trypsin, chymotrypsin, and plasmin) or broad (subtilisin and proteinase K) substrate specificity, applied at very low concentrations, preferentially cleaved both proteins in regions, i.e., subdomains A, B, and C in CSD1 and the proline-rich region (PRR) in MAP2c, that are destined to form contacts with their targets. For CSD1, nonadditivity of the CD spectra of its two halves and suboptimal hydration of the full-length protein measured by solid-state NMR demonstrate that long range tertiary interactions provide the structural background of this structural feature. In MAP2c, such tertiary interactions are absent, which points to the importance of local structural constraints. In fact, urea and temperature dependence of the CD spectrum of its PRR reveals the presence of the extended and rather stiff polyproline II helix conformation that keeps the interaction site exposed. These data suggest that functionally significant residual structure exists in both of these IUPs. This structure, manifest as either transient local and/or global organization, ensures the spatial exposure of short contact segments on the surface. Pertinent data from other IUPs suggest that the presence of such recognition motifs may be a general feature of disordered proteins. To emphasize the possible importance of this structural trait, we propose that these motifs be called primary contact sites in IUPs. PMID- 15751972 TI - Structural basis for thermal stability of human low-density lipoprotein. AB - The stability of human low-density lipoprotein (LDL), the major cholesterol carrier in plasma, was analyzed by heating samples of different concentrations at a rate from 11 to 90 K/h. Correlation of the calorimetric, circular dichroism, fluorescence, turbidity, and electron microscopic data shows that thermal disruption of LDL involves irreversible changes in the particle morphology and protein conformation but no global protein unfolding. Heating to 85 degrees C induces LDL conversion into smaller and larger particles and apparent partial dissociation, but not unfolding, of its sole protein, apoB. Further heating leads to partial unfolding of the beta-sheets in apoB and to fusion of the protein depleted LDL into large aggregated lipid droplets, resulting in a previously unidentified high-temperature calorimetric peak. These lipid droplets resemble in size and morphology the extracellular lipid deposits formed in the arterial wall in early atherosclerosis. The strong concentration dependence of LDL fusion revealed by near-UV/visible CD, turbidity, and calorimetry indicates high reaction order, and the heating rate dependence suggests high activation energy that arises from transient disruption of lipid and/or protein packing interactions in the course of particle fusion and apparent apoB dissociation. Consequently, thermal stability of LDL is modulated by kinetic barriers. Similar barriers may confer structural integrity to LDL subclasses in vivo. PMID- 15751974 TI - Brightness of yellow fluorescent protein from coral (zFP538) depends on aggregation. AB - The yellow fluorescent protein from coral (zFP538) forms aggregates in water solutions. According to dynamic light scattering and gel filtration data, the aggregation number is approximately 1000-10000 at pH 8-9 and protein concentration 1 mg/mL. Gel filtration demonstrated that dissociation of the aggregates takes place upon dilution, and the molecular weight of the aggregates decreases with pH. Atomic force microscopy (AFM) and near-field scanning optical microscopy (NSOM) were used to obtain images of zFP538 in the solid state. It was shown that protein films are comprised of fluorescent ellipsoidal granules with a 50-300 nm major axis and a 30-130 nm minor axis. The dependence of zFP538 fluorescence on protein concentration between 1.2 x 10(-)(9) and 5.5 x 10(-)(7) M can be divided in two linear regions with different slopes indicating the existence of at least two different forms of zFP538. The fluorescence of zFP538 decreases with time upon acidification, and the decrease depends on pH and protein concentration. Between pH 3.5 and pH 5.5, relative residual fluorescence is higher for concentrated zFP538 solutions (about 10(-)(6) M) as compared with diluted ones (10(-)(7) M and below). Aggregation makes zFP538 more stable against fluorescence quenching upon acidification: the decrease in zFP538 fluorescence at protein concentration 1 mg/mL is completely reversible, unlike that observed for less concentrated solutions. This phenomenon may be due to the decrease in the freedom of chromophore mobility in zFP538 aggregates. PMID- 15751973 TI - Exocyclic DNA lesions stimulate DNA cleavage mediated by human topoisomerase II alpha in vitro and in cultured cells. AB - DNA adducts are mutagenic and clastogenic. Because of their harmful nature, lesions are recognized by many proteins involved in DNA repair. However, mounting evidence suggests that lesions also are recognized by proteins with no obvious role in repair processes. One such protein is topoisomerase II, an essential enzyme that removes knots and tangles from the DNA. Because topoisomerase II generates a protein-linked double-stranded DNA break during its catalytic cycle, it has the potential to fragment the genome. Previous studies indicate that abasic sites and other lesions that distort the double helix stimulate topoisomerase II-mediated DNA cleavage. Therefore, to further explore interactions between DNA lesions and the enzyme, the effects of exocyclic adducts on DNA cleavage mediated by human topoisomerase IIalpha were determined. When located within the four-base overhang of a topoisomerase II cleavage site (at the +2 or +3 position 3' relative to the scissile bond), 3,N(4)-ethenodeoxycytidine, 3,N(4)-etheno-2'-ribocytidine, 1,N(2)-ethenodeoxyguanosine, pyrimido[1,2-a]purin 10(3H)-one deoxyribose (M(1)dG), and 1,N(2)-propanodeoxyguanosine increased DNA scission approximately 5-17-fold. Enhanced cleavage did not result from an increased affinity of topoisomerase IIalpha for adducted DNA or a decreased rate of religation. Therefore, it is concluded that these exocyclic lesions act by accelerating the forward rate of enzyme-mediated DNA scission. Finally, treatment of cultured human cells with 2-chloroacetaldehyde, a reactive metabolite of vinyl chloride that generates etheno adducts, increased cellular levels of DNA cleavage by topoisomerase IIalpha. This finding suggests that type II topoisomerases interact with exocyclic DNA lesions in physiological systems. PMID- 15751975 TI - Spectroscopic properties of the carotenoid 3'-hydroxyechinenone in the orange carotenoid protein from the cyanobacterium Arthrospira maxima. AB - The cyanobacterial water-soluble orange carotenoid binding protein (OCP) is an ideal system for study of the effects of protein environment on photophysical properties of carotenoids. It contains a single pigment, the carotenoid 3' hydoxyechinenone (hECN). In this study, we focus on spectroscopic properties of hECN in solution and in the OCP, aiming to elucidate the spectroscopic effects of the carotenoid-protein interaction in the context of the function(s) of the OCP. The noncovalent binding of hECN to the OCP causes a conformational change in the hECN, leading to a prolongation of the effective conjugation length. This change is responsible for shortening of the S(1) lifetime from 6.5 ps in solution to 3.3 ps in the OCP. The conformational change and the hydrogen bonding via the carbonyl group of hECN result in stabilization of an intramolecular charge transfer (ICT) state. No signs of the ICT state were found in hECN in solution, regardless of the solvent polarity; spectral bands in transient absorption spectra of OCP-bound hECN exhibit features typical for the ICT state. Application of global fitting analysis revealed further effects of binding hECN in the OCP. The S(1) state of hECN in the OCP decays with two time constants of 0.9 and 3.3 ps. Modeling of the excited-state processes suggests that these two components are due to two populations of hECN in the OCP that differ in the hydrogen bonding via the carbonyl group. These results support the hypothesis that the OCP functions as a photoprotective shield under excess light. Mechanistically, the broadening of the hECN absorption spectrum upon binding to OCP enhances filtering effect of hECN. Furthermore, the binding-induced conformational change and activation of the ICT state that leads to a shortening of hECN lifetime effectively makes the protein-bound hECN a more effective energy dissipator. PMID- 15751976 TI - Effects of active site mutations on the metal binding affinity, catalytic competence, and stability of the family II pyrophosphatase from Bacillus subtilis. AB - Family II inorganic pyrophosphatases (PPases) have been recently found in a variety of bacteria. Their primary and tertiary structures differ from those of the well-known family I PPases, although both have a binuclear metal center directly involved in catalysis. Here, we examined the effects of mutating one Glu, four His, and five Asp residues forming or close to the metal center on Mn(2+) binding affinity, catalysis, oligomeric structure, and thermostability of the family II PPase from Bacillus subtilis (bsPPase). Mutations H9Q, D13E, D15E, and D75E in two metal-binding subsites caused profound (10(4)- to 10(6)-fold) reductions in the binding affinity for Mn(2+). Most of the mutations decreased k(cat) for MgPP(i) by 2-3 orders of magnitude when measured with Mn(2+) or Mg(2+) bound to the high-affinity subsite and Mg(2+) bound to both the low-affinity subsite and pyrophosphate. In the E78D variant, the k(cat) for the Mn-bound enzyme was decreased 120-fold, converting bsPPase from an Mn-specific to an Mg specific enzyme. K(m) values were less affected by the mutations, and, interestingly, were decreased in most cases. Mutations of His(97) and His(98) residues, which lie near the subunit interface, greatly destabilized the bsPPase dimer, whereas most other mutations stabilized it. Mn(2+), in sharp contrast to Mg(2+), conferred high thermostability to wild-type bsPPase, although this effect was reduced by all of the mutations except D203E. These results indicate that family II PPases have a more integrated active site structure than family I PPases and are consequently more sensitive to conservative mutations. PMID- 15751977 TI - UDP-N-acetylmuramic acid (UDP-MurNAc) is a potent inhibitor of MurA (enolpyruvyl UDP-GlcNAc synthase). AB - Purified recombinant MurA (enolpyruvyl-UDP-GlcNAc synthase) overexpressed in Escherichia coli had significant amounts of UDP-MurNAc (UDP-N-acetylmuramic acid) bound after purification. UDP-MurNAc is the product of MurB, the next enzyme in peptidoglycan biosynthesis. About 25% of MurA was complexed with UDP-MurNAc after five steps during purification that should have removed it. UDP-MurNAc isolated from MurA was identified by mass spectrometry, NMR analysis, and comparison with authentic UDP-MurNAc. Subsequent investigation showed that UDP-MurNAc bound to MurA tightly, with K(d,UDP)(-)(MurNAc) = 0.94 +/- 0.04 microM, as determined by fluorescence titrations using ANS (8-anilino-1-naphthalenesulfonate) as an exogenous fluorophore. UDP-MurNAc binding was competitive with ANS and phosphate, the second product of MurA, and it inhibited MurA. The inhibition patterns were somewhat ambiguous, likely being competitive with the substrate PEP (phosphoenolpyruvate) and either competitive or noncompetitive with respect to the substrate UDP-GlcNAc (UDP-N-acetylglucosamine). These results indicate a possible role for UDP-MurNAc in regulating the biosynthesis of nucleotide precursors of peptidoglycan through feedback inhibition. Previous studies indicated that UDP-MurNAc binding to MurA was not tight enough to be physiologically relevant; however, this was likely an artifact of the assay conditions. PMID- 15751979 TI - Role of hexagonal structure-forming lipids in diadinoxanthin and violaxanthin solubilization and de-epoxidation. AB - In this study, we have examined the influence of different lipids on the solubility of the xanthophyll cycle pigments diadinoxanthin (Ddx) and violaxanthin (Vx) and on the efficiency of Ddx and Vx de-epoxidation by the enzymes Vx de-epoxidase (VDE) from wheat and Ddx de-epoxidase (DDE) from the diatom Cyclotella meneghiniana, respectively. Our results show that the lipids MGDG and PE are able to solubilize both xanthophyll cycle pigments in an aqueous medium. Substrate solubilization is essential for de-epoxidase activity, because in the absence of MGDG or PE Ddx and Vx are present in an aggregated form, with limited accessibility for DDE and VDE. Our results also show that the hexagonal structure-forming lipids MGDG and PE are able to solubilize Ddx and Vx at much lower lipid concentrations than bilayer-forming lipids DGDG and PC. We furthermore found that, in the presence of MGDG or PE, Ddx is much more solubilizable than Vx. This substantial difference in Ddx and Vx solubility directly affects the respective de-epoxidation reactions. Ddx de-epoxidation by the diatom DDE is saturated at much lower MGDG or PE concentrations than Vx de epoxidation by the higher-plant VDE. Another important result of our study is that bilayer-forming lipids DGDG and PC are not able to induce efficient xanthophyll de-epoxidation. Even in the presence of high concentrations of DGDG or PC, where Ddx and Vx are completely solubilized, a strongly inhibited Ddx de epoxidation is observed, while Vx de-epoxidation by VDE is completely absent. This indicates that the inverted hexagonal phase domains provided by lipid MGDG or PE are essential for de-epoxidase activity. We conclude that in the natural thylakoid membrane MGDG serves to solubilize the xanthophyll cycle pigments and furthermore provides inverted hexagonal structures associated with the membrane bilayer, which are essential for efficient xanthophyll de-epoxidase activity. PMID- 15751978 TI - Kinetic isotope effect analysis of the ribosomal peptidyl transferase reaction. AB - The ribosome is the macromolecular machine responsible for protein synthesis in all cells. Here, we establish a kinetic framework for the 50S modified fragment reaction that makes it possible to measure the kinetic effects that result from isotopic substitution in either the A or P site of the ribosome. This simplified peptidyl transferase assay follows a rapid equilibrium random mechanism in which the reverse reaction is nonexistent and the forward commitment is negligible. A normal effect (1.009) is observed for (15)N substitution of the incoming nucleophile at both low and high pH. This suggests that the first irreversible step is the formation of the tetrahedral intermediate. The observation of a normal isotope effect that does not change as a function of pH suggests that the ribosome promotes peptide bond formation by a mechanism that differs in its details from an uncatalyzed aminolysis reaction in solution. This implies that the ribosome contributes chemically to catalysis of peptide bond formation. PMID- 15751980 TI - Efficacy of soluble phospholipids in the prothrombinase reaction. AB - The prothrombinase complex is comprised of an enzyme, factor Xa, and a cofactor, factor Va, that each bind peripherally to membranes containing phosphatidylserine (PS) and activate the substrate, prothrombin. The mechanism by which the membrane contributes to enhanced catalytic efficacy of prothrombinase is not precisely known but is generally attributed to some aspect of enzyme and substrate assembly on the multisite surface of the membrane. A recent proposal has suggested a radically different role in which individual phospholipid molecules, either in the membrane or as single soluble molecules, act by an entirely allosteric mechanism that does not involve the multisite feature of the membrane [Zhai, X., Srivastava, A., Drummond, D. C., Daleke, D., and Lentz, B. R. (2002) Biochemistry 41, 5675-5684]. Our study measured prothrombinse activity in the presence of phospholipids such as short-chain phosphatidylserine and lysophosphatidylserine (lyso-PS). Both enhanced prothrombinase activity, and the increase was consistent with the requirement for extended bilayer structure. Even then, prothrombinase activity was low when compared with activity on bilayer membranes of mixed PS and phosphatidylcholine (PC). Lyso-PS approached the activity of PS/PC membranes only when it was mixed with PC bilayers. The results suggest that the two-dimensional membrane bilayer surface is necessary for the support of full prothrombinase activity. PMID- 15751981 TI - Relationship between the structure of amphiphilic copolymers and their ability to disturb lipid bilayers. AB - Nonionic amphiphiles and particularly block copolymers of ethylene oxide and propylene oxide (Pluronics) cause pronounced chemosensitization of tumor cells that exhibit multiple resistance to antineoplastic drugs. This effect is due to inhibition of P-glycoprotein (P-gp) responsible for drug efflux. It was suggested that the inhibition of P-gp might be due to changes in its lipid surrounding. Indeed, high dependence of P-gp activity on the membrane microviscosity was demonstrated [Regev et al. (1999) Eur. J. Biochem. 259, 18-24], suggesting that the ability of Pluronics to affect the P-gp activity is mediated by their effect on the membrane structure. We have found recently that adsorption of Pluronics on lipid bilayers induced considerable disturbance of the lipid packing [Krylova et al. (2003) Chemistry 9, 3930-3936]. In the present paper, we studied 19 amphiphilic copolymers, including newly synthesized hyperbranched polyglycerols, Pluronic and Brij surfactants, for their ability to accelerate flip-flop and permeation of antitumor drug doxorubicin (DOX) in liposomes. It was found that not only bulk hydrophobicity but also the chemical microstructure of the copolymer determines its membrane disturbing ability. Copolymers containing polypropylene oxide caused higher acceleration of flip-flop and DOX permeation than polysurfactants containing aliphatic chains. The effects of copolymers containing hyperbranched polyglycerol "corona" were more pronounced, as compared to the copolymers with linear poly(ethylene oxide) chains, indicating that a bulky hydrophilic block induces additional disturbances in the lipid bilayer. A good correlation between the copolymer flippase activity and a linear combination of copolymer bulk hydrophobicity and the van der Waals volume of its hydrophobic block was found. The relationship between the structure of a copolymer and its ability to disturb lipid membranes presented in this paper may be useful for the design of novel amphiphilic copolymers capable of affecting the activity of membrane transporters in living cells. PMID- 15751982 TI - Probing the oligomeric state of phospholamban variants in phospholipid bilayers from solid-state NMR measurements of rotational diffusion rates. AB - Phospholamban (PLB) is a small transmembrane protein that regulates calcium transport across the sarcoplasmic reticulum (SR) of cardiac cells. PLB self associates into pentamers within sodium dodecyl sulfate (SDS) micelles, but the oligomeric status of PLB in SR membranes is not known. This work has shown that a mutant of PLB, with all native cysteine residues replaced by alanine (Ala-PLB), runs as a monomer on SDS-PAGE gels, in agreement with previous studies [Karim et al. (2000) Biochemistry 39, 10892-10897]. By contrast, a peptide representing the transmembrane domain of the cysteine-free mutant (TM-Ala-PLB) coexists as pentamers, dimers, and monomers on gels. Solid-state NMR methods were used to examine the size and heterogeneity of Ala-PLB and TM-Ala-PLB labeled with (13)C and (2)H in the transmembrane domain and incorporated into dimyristoylphosphatidylcholine (DMPC) bilayers. Wide line (2)H NMR and (13)C cross-polarization magic-angle spinning (CP-MAS) NMR spectra of Ala-PLB and TM Ala-PLB revealed two distinct species of each of the proteins in the membranes. In the case of Ala-PLB one species was present initially and a second species emerged after 12 h. Measurements of (1)H-(13)C dipolar couplings for the two species of Ala-PLB showed that the rotational diffusion of one species was relatively rapid, defined by a correlation time (tau(R)) of less than 10 micros, whereas the rotation of the other species was comparatively slow (tau(R) approximately 60 micros). These results suggest that although Ala-PLB runs as a monomer on gels, a mixture of different oligomeric forms of the protein, possibly monomers and pentamers, is present in DMPC bilayers. Caution must therefore be exercised in using SDS-PAGE to draw conclusions about the oligomeric state of PLB variants in lipid bilayers. PMID- 15751983 TI - Botulinum neurotoxin serotype F: identification of substrate recognition requirements and development of inhibitors with low nanomolar affinity. AB - Botulinum neurotoxins (BoNTs A-G) are zinc metalloendoproteases that exhibit extraordinary specificities for proteins involved in neurotransmitter release. In view of the extreme toxicities of these molecules, their applications in human medicine, and potential for misuse, it is of considerable importance to elucidate the mechanisms underlying substrate recognition and to develop inhibitors, with the ultimate goal of obtaining anti-botulinum drugs. We synthesized peptides based on vesicle-associated membrane protein (VAMP) to investigate the substrate requirements of BoNT F, which cleaves VAMP between residues Q58 and K59. The minimum substrate was a peptide containing VAMP residues 32-65, which includes only one of the two VAMP structural motifs thought to be required for botulinum substrate recognition. BoNT F exhibited a strict requirement for residues D57 (P(2)), K59 (P(1)'), and L60 (P(2)'), but peptides containing substitutions for R56 (P(3)), Q58 (P(1)), and S61 (P(3)') were cleaved. Therefore, the P(2), P(1)', and P(2)' residues of VAMP are of paramount importance for BoNT F substrate recognition near the scissile bond. K(i) values of uncleavable analogues were similar to K(m) values of the substrate, suggesting that substrate discrimination occurs at the cleavage step, not at the initial binding step. We then synthesized inhibitors of BoNT F that incorporated d-cysteine in place of glutamine 58, exhibited K(i) values of 1-2 nM, and required binding groups on the N-terminal but not the C-terminal side of the zinc ligand. The latter characteristic distinguishes BoNT F from other zinc metalloendoproteases, including BoNTs A and B. PMID- 15751984 TI - A chimeric protein induces tumor cell apoptosis by delivering the human Bcl-2 family BH3-only protein Bad. AB - Deregulation of PI3K/Akt and Raf/Mek/Erk signal transduction cascades is one of the principal causes of neoplastic transformation. The inactivation of the proapoptotic protein Bad, upon phosphorylation by different kinases of these two pathways, may play an important role in different human malignancies. Therefore, we have expressed and purified a new chimeric protein, hGM-CSF-Bad, linking the human granulocyte-macrophage colony-stimulating factor to the N-terminus of the proapoptotic protein human Bad, to deliver Bad into tumor cells and induce apoptosis. Indeed, the human GM-CSF receptor is a good target because it is overexpressed on many leukemias and solid tumors and is not detectable on stem cells. We found that the chimeric protein binds the human GM-CSF receptor, is endocytosed, and appears to reach the cytosol via retrograde ER transport. After entering cells, the protein is able to induce apoptosis of human leukemia cells and human colon and gastric carcinoma cell lines (IC(50) values as low as 1 muM). We conclude that GM-CSF-Bad can overcome the inappropriate survival stimuli in transformed cells and restore the apoptotic pathway. The completely human sequence and the elevated selectivity for cancer cells could prevent immunogenicity and the nonspecific toxicity of targeted toxins in future clinical application of this fusion protein. PMID- 15751985 TI - Expression and characterization of the catalytic domains of soluble guanylate cyclase: interaction with the heme domain. AB - The catalytic domains (alpha(cat) and beta(cat)) of alpha1beta1 soluble guanylate cyclase (sGC) were expressed in Escherichia coli and purified to homogeneity. alpha(cat), beta(cat), and the alpha(cat)beta(cat) heterodimeric complex were characterized by analytical gel filtration and circular dichroism spectroscopy, and activity was assessed in the absence and presence of two different N-terminal regulatory heme-binding domain constructs. Alpha(cat) and beta(cat) were inactive separately, but together the domains exhibited guanylate cyclase activity. Analysis by gel filtration chromatography demonstrated that each of the approximately 25-kDa domains form homodimers. Heterodimers were formed when alpha(cat) and beta(cat) were combined. Results from circular dichroism spectroscopy indicated that no major structural changes occur upon heterodimer formation. Like the full-length enzyme, the alpha(cat)beta(cat) complex was more active in the presence of Mn(2+) as compared to the physiological cofactor Mg(2+), although the magnitude of the difference was much larger for the catalytic domains than for the full-length enzyme. The K(M) for Mn(2+)-GTP was measured to be 85 +/- 18 microM, and in the presence of Mn(2+)-GTP, the K(D) for the alpha(cat)beta(cat) complex was 450 +/- 70 nM. The N-terminal heme-bound regulatory domain of the beta1 subunit of sGC inhibited the activity of the alpha(cat)beta(cat) complex in trans, suggesting a domain-scale mechanism of regulation by NO. A model in which binding of NO to sGC causes relief of an autoinhibitory interaction between the regulatory heme-binding domain and the catalytic domains of sGC is proposed. PMID- 15751986 TI - The P450cam G248E mutant covalently binds its prosthetic heme group. AB - Previous studies on mammalian peroxidases and cytochrome P450 family 4 enzymes have shown that a carboxylic group positioned close to a methyl group of the prosthetic heme is required for the formation of a covalent link between a protein carboxylic acid side chain and the heme. To determine whether there are additional requirements for covalent bond formation in the P450 enzymes, a glutamic acid or an aspartic acid has been introduced into P450(cam) close to the heme 5-methyl group. Spectroscopic and kinetic studies of the resulting G248E and G248D mutants suggest that the carboxylate group coordinates with the heme iron atom, as reported for a comparable P450(BM3) mutant [Girvan, H. M., Marshall, K. R., Lawson, R. J., Leys, D., Joyce, M. G., Clarkson, J., Smith, W. E., Cheesman, M. R., and Munro, A. W. (2004) J. Biol. Chem. 279, 23274-23286]. The two P450(cam) mutants have low catalytic activity, but in contrast to the P450(BM3) mutant, incubation of the G248E (but not G248D) mutant with camphor, putidaredoxin, putidaredoxin reductase, and NADH results in partial covalent binding of the heme to the protein. No covalent attachment is observed in the absence of camphor or any of the other reaction components. Pronase digestion of the G248E P450(cam) mutant after covalent attachment of the heme releases 5 hydroxyheme, establishing that the heme is covalently attached through its 5 methyl group as predicted by in silico modeling. The results establish that a properly positioned carboxyl group is the sole requirement for autocatalytic formation of a heme-protein link in P450 enzymes, but also show that efficient covalent binding requires placement of the carboxyl close to the methyl but in a manner that prevents strong coordination to the iron atom. PMID- 15751987 TI - Effect of modification of the length and flexibility of the acyl carrier protein thioesterase interdomain linker on functionality of the animal fatty acid synthase. AB - A natural linker of approximately 20 residues connects the acyl carrier protein with the carboxy-terminal thioesterase domain of the animal fatty acid synthase. This study examines the effects of changes in the length and amino acid composition of this linker on catalytic activity, product composition, and segmental motion of the thioesterase domain. Deletion of 10 residues, almost half of the interdomain linker, had no effect on either mobility of the thioesterase domain, estimated from fluorescence polarization of a pyrenebutyl methylphosphono moiety bound covalently to the active site serine residue, or functionality of the fatty acid synthase; further shortening of the linker limited mobility of the thioesterase domain and resulted in reduced fatty acid synthase activity and an increase in product chain length from 16 to 18 and 20 carbon atoms. Surprisingly, however, even when the entire linker region was deleted, the fatty acid synthase retained 28% activity. Lengthening of the linker, by insertion of an unusually long acyl carrier protein-thioesterase linker from a modular polyketide synthase, increased mobility of the thioesterase domain without having any significant effect on catalytic properties of the complex. Interdomain linkers could also be used to tether, to the acyl carrier protein domain of the fatty acid synthase, a thioesterase active toward shorter chain length acyl thioesters generating novel short-chain fatty acid synthases. These studies reveal that although truncation of the interdomain linker partially impacts the ability of the thioesterase domain to terminate growth of the acyl chain, the overall integrity of the fatty acid synthase is quite tolerant to moderate changes in linker length and flexibility. The retention of fatty acid synthesizing activity on deletion of the entire linker region implies that the inherent flexibility of the phosphopantetheine "swinging arm" also contributes significantly to the successful docking of the long-chain acyl moiety in the thioesterase active site. PMID- 15751988 TI - Deletions of helices 2 and 3 of human apoA-I are associated with severe dyslipidemia following adenovirus-mediated gene transfer in apoA-I-deficient mice. AB - The objective of this study was to determine the effect of two amino-terminal apolipoprotein A-I (apoA-I) deletions on high-density lipoprotein (HDL) biosynthesis and lipid homeostasis. Adenovirus-mediated gene transfer showed that the apoA-I[Delta(89-99)] deletion mutant caused hypercholesterolemia, characterized by increased plasma cholesterol and phospholipids, that were distributed in the very low density/intermediate density/low-density lipoprotein (VLDL/IDL/LDL) region, and normal triglycerides. The capacity of the mutant protein to promote ATP-binding cassette transporter A1- (ABCA1-) mediated cholesterol efflux and to activate lecithin:cholesterol acyltranserase (LCAT) was approximately 70-80% of the wild-type (WT) control. The phospholipid transfer protein (PLTP) activity of plasma containing the apoA-I[Delta(89-99)] mutant was decreased to 32% of the WT control. Similar analysis showed that the apoA I[Delta(62-78)] deletion mutant in apoA-I-deficient mice caused combined hyperlipidemia characterized by increased triglycerides, cholesterol, and phospholipids in the VLDL/IDL region. There was enrichment of the VLDL/IDL with mutant apoA-I that resulted in reduction of in vitro lipolysis. The capacity of this mutant to promote ABCA1-mediated cholesterol efflux was normal, and the capacity to activate LCAT in vitro was reduced by 53%. The WT apoA-I and the apoA I[Delta(62-78)] mutant formed spherical HDL particles, whereas the apoA I[Delta(89-99)] mutant formed discoidal HDL particles. We conclude that alterations in apoA-I not only may have adverse effects on HDL biosynthesis but also may promote dyslipidemia due to interference of the apoA-I mutants on the overall cholesterol and triglycerides homeostasis. PMID- 15751989 TI - Redox potentials of chlorophylls in the photosystem II reaction center. AB - Water oxidation generating atmospheric oxygen occurs in photosystem II (PSII), a large protein-pigment complex located in the thylakoid membrane. The recent crystal structures at 3.2 and 3.5 A resolutions provide novel details on amino acid side chains, especially in the D1/D2 subunits. We calculated the redox potentials for one-electron oxidation of the chlorophyll a (Chla) molecules in PSII, considering the protein environment in atomic detail. The calculated redox potentials for the dimer Chla (P(D1/D2)) and accessory Chla (Chl(D1/D2)) were 1.11-1.30 V relative to the normal hydrogen electrode at pH 7, which is high enough for water oxidation. The D1/D2 proteins and their cofactors contribute approximately 390 mV to the enormous upshift of 470 mV compared to the redox potential of monomeric Chla in dimethylformamide. The other subunits are responsible for the remaining 80 mV. The high redox potentials of the two accessory Chla Chl(D1/D2) suggests that they also participate in the charge separation process. PMID- 15751990 TI - Ligand binding properties of bacterial hemoglobins and flavohemoglobins. AB - Bacterial hemoglobins and flavohemoglobins share a common globin fold but differ otherwise in structural and functional aspects. The bases of these differences were investigated through kinetic studies on oxygen, carbon monoxide, and nitric oxide binding. The novel bacterial hemoglobins from Clostridium perfringens and Campylobacter jejuni and the flavohemoglobins from Bacillus subtilis and Salmonella enterica serovar Typhi have been analyzed. Examination of the biochemical and ligand binding properties of these proteins shows a clear distinction between the two groups. Flavohemoglobins show a much greater tendency to autoxidation compared to bacterial hemoglobins. The differences in affinity for oxygen, carbon monoxide, and nitric oxide between bacterial hemoglobins and flavohemoglobins are mainly due to differences in the association rate constants. The second-order rate constants for oxygen and carbon monoxide binding to bacterial hemoglobins are severalfold higher than those for flavohemoglobins. A similar trend is observed for NO association with the oxidized iron(III) form of the proteins. No major differences are observed among the values obtained for the dissociation rate constants for the two groups of bacterial proteins studied, and these constants are all rather similar to those for myoglobin. Taken together, our data suggest that differences exist between the mechanisms of ligand binding to bacterial hemoglobins and flavohemoglobins, suggesting different functions in the cell. PMID- 15751991 TI - First reaction of a bare silicon surface with acid chlorides and a one-step preparation of acid chloride terminated monolayers on scribed silicon. AB - Methyl-terminated and acyl chloride terminated monolayers are produced when silicon is scribed under mono- and diacid chlorides, respectively. To the best of our knowledge, this is the first report of the reaction between a bare silicon surface and acid chlorides. This reaction takes place by wetting the silicon surface in the air with the acid chloride and scribing. Scribing activates the silicon surface by removing its passivation layer. We propose that scribed silicon abstracts chlorine from an acid chloride to form an Si-Cl bond and that the resulting acyl radical diffuses back to the surface to condense with the surface and form an alkyl monolayer. X-ray photoelectron spectroscopy (XPS) confirms the presence of chlorine and shows a steady increase in the amount of carbon with increasing alkyl chain lengths of the acid chlorides. Time-of-flight secondary ion mass spectrometry shows SiCl(+) species and a steady increase in representative hydrocarbon fragments with increasing alkyl chain lengths of the acid chlorides. XPS indicates that diacid chlorides react primarily at one of their ends to create acyl chloride terminated surfaces in a single step. The resulting surfaces are shown to react with various amines (piperazine, morpholine, and octylamine) and a protein. Calculations at Hartree-Fock and density functional theory levels are consistent with the proposed mechanism. PMID- 15751992 TI - Organic and inorganic nanoparticle hybrids. AB - Viruses are exemplary models in nanoassembly for their regular geometries, well characterized surface properties, and nanoscale dimensions. Armed with versatile tools aimed at site-directed mutagenesis to modify the virion's surface, conjugation chemistry for capsid coupling, and manipulation of nanoparticles, we have demonstrated nanoscale assembly of inorganic carbon nanotubes and quantum dots with engineered viruses to produce an intimate array of hybrid structures. PMID- 15751994 TI - Magnetic alignment of self-assembled anthracene organogel fibers. AB - High magnetic fields are shown to be remarkably effective to orient self assembled 2,3-bis-n-decyloxyanthracene (DDOA) fibers during organogel preparation. Magnetic orientation of DDOA results in a highly organized material displaying a fiber-orientation order parameter of 0.85, a large linear birefringence, and fluorescence dichroism. The aligned organogel is stable after removal of the magnetic field at room temperature and consists of fibers oriented perpendicular to the magnetic field direction, as shown by scanning electron microscopy. Models for the molecular organization within the gel fibers are discussed upon quantitative analysis of the birefringence. Prospectively, magnetic alignment can be used to improve specific properties of organogel materials. PMID- 15751993 TI - Signal-directed sequential assembly of biomolecules on patterned surfaces. AB - The signal-guided and sequential assembly of biomolecules onto patterned surfaces is demonstrated. Readily transmittable electric signals are used to guide spatially selective deposition of the pH-responsive polysaccharide, chitosan, and functionalized chitosan conjugates, by generating localized pH gradients. The nucleophilic primary amine groups of chitosan enable facile conjugation of proteins and nucleic acids by two approaches, one an enzymatic approach and the other a standard chemical modification, thus providing flexibility when sequentially assembling biomolecules in a spatially selective manner. Moreover, we developed an agarose gel "biomask" for the sequential assembly of single stranded DNA and confirmed its functionality through nucleic acid hybridization assays. PMID- 15751995 TI - Controlled synthesis of nonspherical microparticles using microfluidics. AB - The controlled synthesis of nonspherical microparticles using microfluidics processing is described. Polymer droplets, formed by shearing a photopolymer using a continuous water phase at a T-junction, were constrained to adopt nonspherical shapes by confining them using appropriate microchannel geometries. Plugs were obtained by shearing the polymer phase at low shear rates, while disks were obtained by flattening droplets using a channel of low height. The nonspherical shapes formed were permanently preserved by photopolymerizing the constrained droplets in situ using ultraviolet light. Monodisperse plugs and disks of different lengths and diameters were obtained by varying the flow rates of the two phases. PMID- 15751996 TI - Altering the crystal morphology of silicalite-1 through microemulsion-based synthesis. AB - The crystal morphology of silicalite-1 was adjusted through a microemulsion-based hydrothermal synthesis. The surfactant cetyltrimethylammonium bromide (CTAB) with cosurfactant butanol was used to form water-in-oil microemulsions containing the silicalite-1 synthesis gel. The crystal morphology of silicalite-1 was adjusted from coffin-shaped to novel rod-shaped and to irregular-shaped nanoparticles by varying the microemulsion composition. Silicalite-1 synthesized in the microemulsion has a smaller size and a more narrow size distribution than that produced by conventional synthesis without the microemulsion. The novel morphology of silicalite-1 may facilitate assembly into films and find applications in separation and catalysis. PMID- 15751997 TI - Pd nanoparticle aging and its implications in the suzuki cross-coupling reaction. AB - Examination of the catalysts recovered in the N,N-dihexylcarbodiimide-palladium nanoparticle composite catalyzed Suzuki cross-coupling reactions revealed that the metal nanoparticles transformed gradually from spherical-shape to larger needle-shaped crystals. Two types of Ostwald ripening processes were observed. One involves rapid aggregation of the incipient nanoparticle catalyst (2-5 nm) into blackberry-like assemblies (100-200 nm), which is accompanied with the much slower dissolution of small crystals or amorphous nanoparticles and the formation of larger needle-shaped crystals. The observed structural changes provided new insights into the durability of the polymer nanoparticle composite catalyst. PMID- 15751998 TI - Synthesis of SiO2/polystyrene nanocomposite particles via miniemulsion polymerization. AB - The SiO(2)/polystyrene nanocomposite particles were synthesized through miniemulsion polymerization by using sodium lauryl sulfate surfactant (SLS), hexadecane costabilizer in the presence of silica particles coated with methacryloxy(propyl)trimethoxysilane. Core-shell or other interesting morphology composite particles were obtained depending on the size of the silica particles and the surfactant concentration employed. By adjusting these parameters, it was possible to control the size and morphology of the composite particles. PMID- 15751999 TI - A variable-temperature diffuse reflectance infrared fourier transform spectroscopy study of the binding of water and pyridine to the surface of acid activated metakaolin. AB - Four metakaolins were prepared by heating a Spanish kaolin at 600, 700, 800, and 900 degrees C for 10 h. Following preliminary optimization, these metakaolins were acid activated in 6 M hydrochloric acid at 90 degrees C for 6 h; the samples calcined at 600, 700, and 800 degrees C produced the highest surface area solids and were selected for further study. Variable-temperature diffuse reflectance infrared Fourier transform spectroscopy analysis of the resulting acid-activated metakaolins (AAMKs) identified a wide range of hydrogen bond strengths in adsorbed water at room temperature. Above 300 degrees C it was possible to fit the broad hydroxyl stretching band to seven contributing components at 3730, 3700, 3655, 3615, 3583, 3424, and 3325 cm(-1). As the sample temperature was increased, the 3730 cm(-1) band increased in intensity as the water hydrogen bonded to AlOHAl was thermally desorbed. The other six bands decreased in intensity. The spectra of adsorbed pyridine indicated the presence of both Bronsted and Lewis acid sites on the surface of the air-dried AAMKs. Preheating the AAMK at 200 degrees C prior to pyridine sorption reduced the number of Bronsted acid sites and increased the number of thermally stable Lewis acid sites. A reduction in the amount of adsorbed pyridine after pretreating the AAMK at 400 degrees C was tentatively attributed to a reduction in surface area. This was reflected in fewer thermally stable Lewis acid sites in the AAMK pretreated at 400 degrees C compared to the number present in the sample pretreated at 200 degrees C. PMID- 15752000 TI - Structure and interactions of block copolymer micelles of Brij 700 studied by combining small-angle X-ray and neutron scattering. AB - Spherical micelles of the diblock copolymer/surfactant Brij 700 (C(18)EO(100)) in water (D(2)O) solution have been investigated by small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS). SAXS and SANS experiments are combined to obtain complementary information from the two different contrast conditions of the two techniques. Solutions in a concentration range from 0.25 to 10 wt % and at temperatures from 10 to 80 degrees C have been investigated. The data have been analyzed on absolute scale using a model based on Monte Carlo simulations, where the micelles have a spherical homogeneous core with a graded interface surrounded by a corona of self-avoiding, semiflexible interacting chains. SANS and SAXS data were fitted simultaneously, which allows one to obtain extensive quantitative information on the structure and profile of the core and corona, the chain interactions, and the concentration effects. The model describes the scattering data very well, when part of the EO chains are taken as a "background"contribution belonging to the solvent. The effect of this becomes non-negligible at polymer concentrations as low as 2 wt %, where overlap of the micellar coronas sets in. The results from the analysis on the micellar structure, interchain interactions, and structure factor effects are all consistent with a decrease in solvent quality of water for the PEO block as the theta temperature of PEO is approached. PMID- 15752001 TI - Release and formation of surface-localized ionic clusters (SLICs) into phospholipid rafts from colloidal solutions during coalescence. AB - Stimuli-responsive behavior of phospholipids in the presence of ionic surfactants utilized in synthesis of MMA/nBA colloidal particles was investigated. Utilizing 1-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC) phospholipid, and sodium dioctyl sulfosuccinate (SDOSS) surfactant as dispersing media in H(2)O, narrow unimodal particle size distributions of methyl methacrylate (MMA)/n-butyl acrylate (nBA) copolymers were synthesized. The particle diameters were 154 nm when a SDOSS/MHPC mixture was used and 161 nm using MHPC as the only surface stabilizing species. When such colloidal dispersions are exposed to 1.7, 3.3, and 6.7 mM aqueous CaCl(2) and KCl electrolyte solutions, surface-localized ionic clusters are generated at the film-air interface that may serve as lipid rafts composed of crystalline phases of MHPC deposited on poly(MMA)/nBA films. These studies illustrate that it is possible to control release and morphology developments of surface phospholipid rafts on artificial surfaces. PMID- 15752002 TI - Pickering emulsions with controllable stability. AB - We prepare solid-stabilized emulsions using paramagnetic particles at an oil/water interface that can undergo macroscopic phase separation upon application of an external magnetic field. A critical field strength is found for which emulsion droplets begin to translate into the continuous-phase fluid. At higher fields, the emulsions destabilize, leading to a fully phase-separated system. This effect is reversible, and long-term stability can be recovered by remixing the components with mechanical agitation. PMID- 15752003 TI - Porphyrin-based organogels: control of the aggregation mode by a pyridine carboxylic acid interaction. AB - To find a new strategy for designing porphyrin-based organogelators, hydrogen bond-donating (carboxylic acid)/accepting (pyridine) substituents or electron donating (dialkylamino)/withdrawing (pyridine) substituents were introduced into peripheral positions of a porphyrin (1cp or 1ep, respectively), and the gelation properties were compared with those of symmetrical reference compounds bearing two pyridyl substituents or two ester groups (1pp or 1ee, respectively). It was found that the symmetrical molecules show a solubility that is quite inferior to that of 1cp and 1ep having a dipole moment and precipitate from most organic solvents. 1cp, 1ep, and 1ee formed gels with cyclohexane, methylcyclohexane, and several alcoholic solvents, but scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observations revealed that their superstructures constructed in the organogels are very different. In cyclohexane, 1cp resulted in a sheetlike structure, whereas 1ep and 1ee resulted in a fiberlike structure. The difference is attributed to the two-dimensional interactive forces in 1cp consisting of the porphyrin-porphyrin pi-pi stacking and the carboxylic acid-pyridine hydrogen bonding. In fact, when the hydrogen bonding interaction was weakened by alcoholic solvents or by adding pyridine or N,N-(dimethylamino)pyridine, the sheetlike structure was transfigured to the fiberlike structure. Further detailed analyses of their aggregation modes were conducted by spectroscopic methods such as ultraviolet-visible (UV-vis) absorption, Fourier transform infrared (FT-IR), and X-ray diffraction (XRD). On the basis of these findings, the influence of these peripheral substituents on the gel formation and the aggregation mode was discussed. PMID- 15752004 TI - Colloid filtration theory and the Happel sphere-in-cell model revisited with direct numerical simulation of colloids. AB - The transport of colloids and bacterial cells through saturated porous media is a complex phenomenon involving many interrelated processes that are often treated via application of classical colloid filtration theory (CFT). This paper presents a numerical investigation of CFT from the Lagrangian perspective, to evaluate the role of some of the classical assumptions underlying the theory and to demonstrate a means to include processes relevant to bacterial transport that were inadequately characterized or neglected in the original formulation, including Brownian diffusion and potentially hysteretic potential functions. The methodology is based on conducting a Lagrangian trajectory analysis within Happel's sphere-in-cell porous media model to obtain the collection efficiency (eta), the frequency at which colloids or bacteria make contact with the solid phase of the porous medium. The Lagrangian framework of our model lends itself to mechanistic modeling of the biological processes that may be important in subsurface bacterial transport. The numerical study presented here focuses on the size range of bacterial colloids and smaller (down to 10 nm). Results of our model runs are in good agreement with the deterministic trajectory analysis of Rajagopalan and Tien (when diffusion is neglected) and in excellent agreement with the analytical solution to the Smoluchowski-Levich approximation of the convective-diffusion equation (when external forces and interception are neglected). Simple addition of our result for the deterministic eta to our result for the Smoluchowski-Levich eta matches the overall Rajagopalan and Tien eta to within 5% error or less for all cases studied. When we simulate diffusion and the deterministic forces together, our results diverge from the Rajagopalan and Tien eta as the particle size decreases, with discrepancies as large as 73%. These results suggest that accurate prediction of eta values for bacteria-sized (and all submicrometer) colloids requires simultaneous consideration of the primary transport mechanisms. PMID- 15752005 TI - Polyelectrolyte-induced peeling of charged multilamellar vesicles. AB - We study mixtures of charged surfactants, which alone in solution form uni- and multilamellar vesicles, and oppositely charged polyelectrolytes (PEs). The phase behavior is investigated at fixed surfactant concentration as a function of the PE-to-surfactant charge ratio, x. We find that, for x > 0, aggregates form. Light microscopy and X-ray scattering experiments show that the isoelectric point plays a crucial role, since the morphology and the microscopic structure of the aggregates are different before (x < or = 1) and after the isoelectric point (x > 1). To better understand the dynamics for the formation of PE/surfactant complexes, we perform light microscopy experiments where we follow in real time the effect of a PE solution on one multilamellar vesicle (MLV). We find that the PE induces a peeling of the bilayers of the MLV one by one. The peeling is accompanied by strong shape fluctuations of the MLV and leads ultimately to a pile of small aggregates. This novel phenomenon is analyzed in detail and discussed in terms of PE-induced tension and pore formation and growth in a surfactant bilayer. PMID- 15752006 TI - On the surface conductance, flow rate, and current continuities of microfluidics with nonuniform surface potentials. AB - The characteristics of electrokinetic flow in a microchannel depend on both the nature of surface potentials, that is, whether it is uniform or nonuniform, and the electrical potential distribution along the channel. In this paper, the nonlinear Poisson-Boltzmann equation is used to model the electrical double layer and the lattice Boltzmann model coupled with the constraint of current continuity is used to simulate the microfluidic flow field in a rectangular microchannel with a step variation of surface potentials. This current continuity, including surface conduction, convection, and bulk conduction currents, has often been neglected in the literature for electroosmotic flow with nonuniform (heterogeneous) microchannels. Results show that step variation of ion distribution caused by step variation surface potential will influence significantly the electrical potential distribution along the channel and volumetric flow rate. For the system considered, we showed that the volumetric flow rate could have been overestimated by as much as 70% without consideration of the current continuity constraint. PMID- 15752007 TI - Atomic force microscopy study of the interaction between adsorbed poly(ethylene oxide) layers: effects of surface modification and approach velocity. AB - The interaction forces between layers of the triblock copolymer Pluronic F108 adsorbed onto hydrophobic radio frequency glow discharge (RFGD) thin film surfaces and hydrophilic silica, in polymer-free 0.15 M NaCl solution, have been measured using the atomic force microscope (AFM) colloid probe technique. Compression of Pluronic F108 layers adsorbed on the hydrophobic RFGD surfaces results in a purely repulsive force due to the steric overlap of the layers, the form of which suggests that the PEO chains adopt a brush conformation. Subsequent fitting of these data to the polymer brush models of Alexander-de Gennes and Milner, Witten, and Cates confirms that the adsorbed Pluronic F108 adsorbs onto hydrophobic surfaces as a polymer brush with a parabolic segment density profile. In comparison, the interaction between Pluronic F108 layers adsorbed on silica exhibits a long ranged shallow attractive force and a weaker steric repulsion. The attractive component is reasonably well described by van der Waals forces, but polymer bridging cannot be ruled out. The weaker steric component of the force suggests that the polymer is less densely packed on the surface and is less extended into solution, existing as polymeric isolated mushrooms. When the surfaces are driven together at high piezo ramp velocities, an additional repulsive force is measured, attributable to hydrodynamic drainage forces between the surfaces. In comparing theoretical predictions of the hydrodynamic force to the experimentally obtained data, agreement could only be obtained if the flow profile of the aqueous solution penetrated significantly into the polymer brush. This finding is in line with the theoretical predictions of Milner and provides further evidence that the segment density profile of the adsorbed polymer brush is parabolic. A velocity dependent additional stepped repulsive force, reminiscent of a solvation oscillatory force, is also observed when the adsorbed layers are compressed under high loads. This additional force is presumably a result of hindered drainage of water due to the presence of a high volume fraction of polymer chains between the surfaces. PMID- 15752008 TI - Grafting of amino functional monomer onto initiator-modified polystyrene particles. AB - Polystyrene nanoparticles with grafted chains of an amino functionalized polymer were prepared by a two-step polymerization process. In the first step, the polystyrene seed particles were synthesized by the conventional batch emulsion polymerization using terpolymer HAS (hydroperoxide monomer, acrylic acid, and styrene) as a surface-active initiator. The surface of the obtained particles contains carboxyl groups, which are responsible for the latex stability, and residual undecomposed hydroperoxide groups. Therefore, in the second step, an amino functional monomer was grafted onto the hydroperoxide modified polystyrene particles by a "grafting from" approach. X-ray photoelectron spectroscopy, NMR, and scanning electron microscopy were used to examine the surface of the amino functionalized particles. The amount of incorporated amino groups onto the particles was determined by fluorescenometric titration. In general, the number of amino groups on the particle surface increased with the increase of the functional monomer content in the reaction mixture. The incorporation of the functional monomer was also confirmed by electrophoretic measurements. Final particles possess amphoteric character due to the presence of amino and carboxyl groups on the surface. Adsorption of human immunoglobulins G onto the amino functionalized particles was studied as a function of pH and ionic strength. The covalent binding of human IgG was performed using the glutaraldehyde preactivation method. The immunoreactivity of the latex-IgG complex was examined by the latex agglutination test. PMID- 15752010 TI - In situ investigations on organic foam films using neutron and synchrotron radiation. AB - We report on small-angle neutron scattering (SANS) and X-ray scattering (SAXS) investigations of foam films stabilized by sodium dodecyl sulfate. Previous measurements on dry foams (Axelos, M. A. V.; Boue, B. Langmuir 2003, 19, 6598) have shown the presence of spikes in the two-dimensional scattering data which suggest that the incident beam is reflected on some film surfaces. The latter interpretation is confirmed by new neutron studies performed on ordered ("bamboo") foams which allow selection of single films. In the first case, we show that the spikes of the scattered intensity can be obtained by reflection on two parts of the foam, namely, the films and the Plateau borders. With synchrotron radiation, first observations of distinct interference fringes have allowed an accurate determination of the film thickness. A comparison with X-ray and neutron data is made, opening a general discussion about the capabilities of small-angle scattering techniques for studying the microscopic properties of foam films. PMID- 15752009 TI - Formation and characterization of phospholipid monolayers spontaneously assembled at interfaces between aqueous phases and thermotropic liquid crystals. AB - This paper reports an experimental investigation of the self-assembly of phospholipids (l-alpha-phosphatidylcholine-beta-oleoyl-gamma-palmitoyl (l-POPC), dipalmitoyl phosphatidylcholine (DPPC), and l-alpha-dilauroyl phosphatidylcholine (l-DLPC)) at interfaces between aqueous phases and the nematic liquid crystal (LC) 4'-pentyl-4-cyanobiphenyl. Stable planar interfaces between the aqueous phases and LCs were created by hosting the LCs within gold grids (square pores with widths of 283 microm and depths of 20 microm). At these interfaces, the presence and lateral organization of the phospholipids leads to interface-driven orientational transitions within the LC. By doping the phospholipids with a fluorescently labeled lipid (Texas Red-1,2-dipalmitoyl-sn-glycero-3 phosphoethanolamine (TR-DPPE)), quantitative epifluorescence microscopy revealed the saturation coverage of phospholipid at the interface to be that of a monolayer with an areal density of approximately 49 +/- 8% relative to hydrated lipid bilayers. By adsorbing phospholipids to the aqueous-LC interface from either vesicles or mixed micelles of dodecyltrimethylammonium and phospholipid, control of the areal density of phospholipid from 42 +/- 10 to 102 +/-18% of saturation monolayer coverage was demonstrated. Fluorescence recovery after photobleaching (FRAP) experiments performed by using laser scanning confocal microscopy (LSCM) revealed the lateral mobility of fluorescently labeled DPPE in l-DLPC assembled at the interface with the liquid crystal to be (6 +/- 1) x 10( 12) m(2)/s for densely packed monolayers. Variation of the surface coverage and composition of phospholipid led to changes in lateral diffusivity between (0.2 +/ 0.1) x 10(-12) and (15 +/- 2) x 10(-12) m(2)/s. We also observed the phospholipid-laden interface to be compartmentalized by the gold grid, thus allowing for the creation of patterned arrays of phospholipids at the LC-aqueous interface. PMID- 15752011 TI - Role of desorption kinetics in determining marangoni flows generated by using electrochemical methods and redox-active surfactants. AB - We report quantitative measurements of Marangoni flows generated at the surfaces of aqueous solutions by using water-soluble redox-active surfactants in combination with electrochemical methods. These measurements are interpreted within the framework of a simple model that is based on lubrication theory and the proposition that the kinetics of the desorption of redox-active surfactants from the surfaces of aqueous solutions plays a central role in determining the strength of the Marangoni flow. The model predicts that the leading edge velocity of the Marangoni flow will decay exponentially with time and that the rate constant for the decay of the velocity can yield an estimate of the surfactant desorption rate constant. Good agreement between theory and experiments was found. By interpreting experimental measurements of electrochemically generated Marangoni flows within the framework of the model, we conclude that the desorption rate constant of the redox-active surfactant Fc(CH(2))(11) N(+)(CH(3))(3)Br(-), where Fc is ferrocene, is 0.07 s(-)(1). We also conclude that the ionic strength of the aqueous solution has little effect on the desorption rate constant of the ferrocenyl surfactant. PMID- 15752012 TI - Anion effects on calixarene monolayers: a Hofmeister series study. AB - Due to their amphiphilic structure, calixarenes adsorb at the air/water interface and form stable Langmuir films. We have explored the effect of salts on calix[6]- and calix[8]arene spreading isotherms at the air/water interface. A wide range of different potassium salts was used in the subphase: KCl, KI, KBr, KSCN, KNO(3), CH(3)COOK, K(2)SO(4), and K(3)PO(4). The differences in Langmuir isotherms are due to the presence of different anions in the subphase, to the different conformations of the ligands at the interface, and to the different complexing affinities of calix[6]- and calix[8]arene for potassium ions. The two systems show a significant specific ion effect that can be discussed in terms of Hofmeister series. Characteristic monolayer parameters, e.g., limiting area (A(lim)), collapse pressure (pi(coll)), modulus of compressibility (C(s)(-1)), and surface potential (DeltaV), are discussed in terms of some physicochemical parameters that reflect dispersion forces: in particular, anion polarizabilities, lyotropic number (N), molar surface tension increment (sigma), and partial molar volume (nu(s)). PMID- 15752013 TI - Blends of amphiphilic poly(dimethylsiloxane) and nonamphiphilic octaisobutyl-POSS at the air/water interface. AB - Brewster angle microscopy (BAM) shows that a nonamphiphilic polyhedral oligomeric silsesquioxane (POSS) nanofiller, octaisobutyl-POSS, forms aggregates at all surface concentrations at the air/water interface. When amphiphilic poly(dimethylsiloxane) (PDMS) is blended with the octaisobutyl-POSS (>10 wt % PDMS), the degree of POSS aggregation dramatically decreases. Thermodynamic analyses and morphology studies through surface pressure-area per monomer isotherm data and BAM, respectively, exhibit three distinct composition regimes: (1) Blends with >70 wt % POSS have unstable isotherms whose shapes deviate from those of PDMS and form large rigid domains comparable to but smaller than pure, octaisobutyl-POSS films. (2) At compositions between approximately 40 and 70 wt % POSS, the isotherms' features are qualitatively similar to those of pure PDMS, and extensive nanofiller "networks" are observed by BAM. (3) For compositions < or = approximately 30 wt % POSS, the isotherms are essentially those of pure PDMS with small POSS domains dispersed in the PDMS matrix. These results provide further insight into nanofiller aggregation mechanisms and dispersion that may be present in thicker films and bulk systems. PMID- 15752014 TI - Molecular orientation of evaporated pentacene films on gold: alignment effect of self-assembled monolayer. AB - Pentacene films deposited on self-assembled monolayers (SAMs) bearing different terminal functional groups have been studied by reflection-absorption IR, grazing angle XRD, NEXAFS, AFM, and SEM analyses. A film with pentacene molecules nearly perpendicularly oriented was observed on Au surfaces covered with an SAM of alkanethiol derivative of X-(CH2)(n)-SH, with X = -CH(3), -COOH, -OH, -CN, NH(2), C(60), or an aromatic thiol p-terphenylmethanethiol. On the other hand, a film with the pentacene molecular plane nearly parallel to the substrate surface was found on bare Au surface. A similar molecular orientation was found in thinner ( approximately 5 nm) and thicker (100 nm) deposited films. Films deposited on different surfaces exhibit distinct morphologies: with apparently smaller and rod-shaped grains on clean bare Au surface but larger and islandlike crystals on SAM-modified surfaces. X-ray photoemission electron microscopy (X PEEM) was used to analyze the orientation of pentacene molecules deposited on a SAM-patterned Au surface. With the micro-NEXAFS spectra and PEEM image analysis, the microarea-selective orientation control on Au was characterized. The ability to control the packing orientation in organic molecular crystals is of great interest in fabricating organic field effect transistors because of the anisotropic nature of charge transport in organic semiconducting materials. PMID- 15752015 TI - Simulated adsorption properties and synthesis prospects of homochiral porous solids based on their heterochiral analogs. AB - Molecular simulations of chiral molecules in porous heterochiral materials were performed to investigate fundamental adsorption properties and possibilities for production of homochiral porous solids. Zeolite BEA polymorph A and zeotype UCSB 7K each provide separated pores of opposite chirality. Single enantiomer and racemic mixture adsorption results are presented and indicate that significant equilibrium enantiomeric excesses of 40-70% in UCSB-7K and 10% in BEA can be achieved. Larger, better-fitting molecules display higher enantiomeric excesses. For dimethylallene, which moves on molecular dynamics time scales in UCSB-7K, self-diffusivities vary by almost an order of magnitude between the two opposite handed UCSB-7K pores for a given enantiomer. The predicted properties indicate that equilibrium and nonequilibrium strategies using related homochiral materials for separations may be successful. To this end, a discussion of strategies for selectively blocking pores of one chirality on the basis of enantiomer segregation is provided. PMID- 15752016 TI - Decontamination of gaseous acetaldehyde over CoOx-loaded SiO2 xerogels under ambient, dark conditions. AB - A series of CoO(x)-doped silica xerogels with various Co(2+) loadings (Co/Si = 0, 1, 2, 4, 6, and 10 mol %) has been prepared. All xerogels exhibit large (800-1050 m(2)/g) surface areas. Narrow pore size distributions with pore size maxima around 3 nm are characteristic for Co/Si = 1, 2, 4, 6, 10 samples. As-prepared CoO(x)/SiO(2) xerogels show high catalytic activity in the air oxidation of gaseous acetaldehyde at room temperature. Carbon dioxide and trace amounts of methane are the only products detected in the gas phase. Acetic acid, a less volatile product, resides on the surface of the xerogels but can slowly desorb. The formation of CO(2) begins after an induction period. The beginning of CO(2) production coincides with the conversion of Co(2+) incorporated in the SiO(2) framework into Co(3+). Thermogravimetry/gas chromatography/mass spectrometry analysis, UV-vis and FTIR spectroscopies, as well as kinetic measurements are employed for CoO(x)/SiO(2) catalyst characterization. A possible mechanism of the reaction is discussed. PMID- 15752017 TI - An experimental study of gas adsorption on fractal surfaces. AB - The validity of the fractal versions of the FHH and BET theories for describing the adsorption of butane and nitrogen on a variety of partially dehydroxylated silica surfaces has been tested. The fractal dimensions obtained from adsorption data have been compared with those obtained completely independently using SAXS. It was found that the fractal dimensions obtained from butane adsorption isotherms, using both the fractal FHH and fractal BET theories, agreed well with the corresponding values obtained from SAXS over overlapping length scales. However, in general, a systematic deviation between the fractal dimension obtained from nitrogen adsorption and that obtained from SAXS was observed. The fractal dimensions obtained from nitrogen adsorption were consistently larger than those obtained from SAXS, which is the opposite of what has often been found in the literature. It has been suggested that the differences in the suitability of the adsorption theories tested to describe butane and nitrogen adsorption is due to the significant difference between the interaction strengths of these two different molecules with silica surfaces. A modified theory that can account for the discrepancy between the fractal dimensions obtained from nitrogen adsorption and SAXS has been proposed. The implications of the new theory for the accuracy of nitrogen adsorption BET surface areas for silicas are discussed. PMID- 15752018 TI - Effects of pulsed low-frequency electromagnetic fields on water characterized by light scattering techniques: role of bubbles. AB - Well-characterized purified water was exposed for 6 h to pulsed low-frequency weak electromagnetic fields. After various time periods, nondegassed and degassed water samples were analyzed by static light scattering. Just after electromagnetic exposure (day 0), a reduction of over 20% in the maximum light scattering intensity at 488 nm wavelength in both nondegassed and degassed samples was observed. By contrast, on day 12 the difference was observed only in nondegassed water samples. The latter effect was attributed to the different geometries of the containers combined with the basic origin of the whole phenomenon due to gas bubbles present in water. By the use of dynamic light scattering, the bubble mean diameter was estimated to be around 300 nm. Our results suggest that the electromagnetic exposure acts on gas nanobubbles present in water and emphasizes the role of the gas/liquid interface. The possibility that exposure to electromagnetic fields disturbs the ionic double layer that contributes to bubble stabilization in water is discussed. PMID- 15752019 TI - Oriented monolayers prepared from lyotropic chromonic liquid crystal. AB - We use a layer-by layer electrostatic self-assembly technique to obtain in-plane oriented aggregates of mesogenic dye molecules cast from lyotropic chromonic liquid crystals (LCLCs) on mica substrates. The aqueous solutions of dye used for deposition are in the nematic phase. Atomic force microscopy and X-ray photoelectron spectroscopy of the dried film reveal that the LCLC molecules adsorb at the charged substrate preserving ordered aggregates of elongated shape characteristic of the nematic phase in the aqueous solution. These elongated aggregates of LCLC molecules form films with in-plane orientational order and are compositionally distinct from the substrate. PMID- 15752020 TI - Self-assembly of the perfluoroalkyl-alkane F14H20 in ultrathin films. AB - Scanning force microscopy on monomolecular films of eicosylperfluorotetradecane, F(CF(2))(14)(CH(2))(20)H, on mica, silicon oxide, or water revealed spontaneous organization to well-defined nanoscopic ribbon and spiral or toroidal superstructures. Whether ribbons or nanospirals were formed depended on the solvent from which the molecular monofilm was cast. Ribbons were observed when a hydrocarbon or a perfluorocarbon solvent was used, e.g., decalin or perfluorodecalin. When the compound, however, was deposited from nonselective hexafluoroxylene, the molecules assembled into spirals of defined size. The spirals/toroids transformed to ribbons when exposed either to decalin or perfluorodecalin vapor, and the ribbons transformed to toroids when exposed to hexafluoroxylene vapor. These changes could be observed in situ. Scanning force microscopy yielded an identical height and width for the bands forming the spirals and for the parallel flat ribbons. X-ray reflectivity yielded a height of 3.61 +/- 0.05 nm, again identical for both morphologies. Yet, the length of the extended F(CF(2))(14)(CH(2))(20)H molecule, i.e., 4.65 nm, exceeds the layer thickness obtained from X-ray reflectometry. It is, however, consistent with an arrangement where the fluorinated chains are oriented normal to the surface layer and where the alkyl segments are tilted with a 122 degrees angle between the two segments. Within the plane defined by the tilt, this angle allows a dense packing of the alkyl segments compensating for the larger cross-section of the fluorocarbon segment. The tilt plane defines an "easy" direction along which the monolayer structure can preserve order. In the plane perpendicular to this axis, long-range ordered dense packing of the alkyl chains is not possible. Incommensurable packing can in principle explain the finite and regular width of the ribbons and the stepwise turn in the spirals. PMID- 15752021 TI - In situ study of the thermoresponsive behavior of micropatterned hydrogel films by imaging ellipsometry. AB - A patterned hydrogel was immobilized on a polymer substrate by low-pressure argon plasma treatment using a masking technique. The polymer sample showed a thermoresponsive aggregation behavior in the region of 35-37 degrees C. The micropatterned, thermoresponsive hydrogel film has been characterized with imaging ellipsometry. The characterization was carried out on the dry film as well as on a swollen sample in water. The thermoresponsive behavior was studied in deionized water by temperature-dependent measurements in a solid-liquid cell. Through imaging ellipsometry, it was possible to distinguish the different regions of interest on a micrometer scale and to follow the swelling of the hydrogel part as a function of the temperature. It was possible to visualize the swelling as 3D profiles of Delta at various temperatures. Long-term changes of the sample could also be detected, which cannot be picked up by conventional ellipsometry. PMID- 15752022 TI - Compression behavior of Langmuir-Blodgett monolayers of regioselectively substituted cellulose ethers with long alkyl side chains. AB - Regioselectively substituted alkylcellulose ethers having long alkyl side chains, 6-O- (6C18), 2,3-di-O- (23C18), and tri-O-octadecyl-cellulose (triC18) were successfully synthesized. The key step of these syntheses was removal of the residual alkylation reagent by an isothermal crystallization procedure to isolate and purify the compounds, since a physical entanglement between the long alkyl side chains in the cellulose derivatives and the reagent had caused difficulty in obtaining the purified derivatives. After the monolayers from these cellulose ethers were fabricated on a water surface, they were deposited on substrates by a vertical dipping method to be Langmuir-Blodgett (LB) monolayers. During the compression process of each monolayer, a surface pressure (pi)-area (A) isotherm behaved in a different way. Atomic force microscopy (AFM) was employed to interpret changes of the surface topography of the obtained LB monolayers depending on the surface pressure. The compressed 23C18 LB monolayer was observed to be more homogeneous than other samples. On the basis of the LB monolayer thickness estimated by AFM as well as X-ray reflection measurements, the 23C18 LB monolayer was assumed to possibly possess the vertical arrangement of an up ordering of all the alkyl side chains on the individual glucose ring against the water surface. In other words, with increase in the surface pressure, the usual conformation of a 2(1) screw of cellulose backbone may be changed into an unusual conformation with a certain phi-psi dihedral angle resulting in 1-fold axis without a symmetry element. These results suggest that the formation of such compressed LB monolayers was strongly influenced by the hydrophobic interaction among the distribution of the long alkyl side chains in the anhydroglucose unit and further lack of inter- and/or intramolecular hydrogen bonds engaged in cellulose ethers, and as a result, those effects may even change the main chain conformation. PMID- 15752023 TI - Particle zips: vertical emulsion films with particle monolayers at their surfaces. AB - Vertical emulsion films with particle monolayers at their surfaces have been studied by direct microscope observations. The effects of particle wettability and surface coverage on the structure and stability of water films in octane and octane films in water have been investigated. Monodisperse silica particles (3 microm in diameter) hydrophobized to different extents have been used. It is found that the structure and stability of emulsion films strongly depend on the film type (water-in-oil or oil-in-water), the particle contact angle, the interactions between particles from the same and the opposite monolayer, and the monolayer density. Stable films are observed only when the particle wettability fulfills the condition for stable particle bridges--in agreement with the concept that hydrophilic particles can give stable oil-in-water emulsions, whereas hydrophobic ones give water-in-oil emulsions. In the case of water films with dilute disordered monolayers at their surfaces, the hydrophilic particles are expelled from the film center toward its periphery, giving a dimple surrounded by a ring of particles bridging the film surfaces. In contrast, the thinning of octane films with dilute ordered monolayers at their surfaces finally leads to the spontaneous formation of a dense crystalline monolayer of hydrophobic particles bridging both surfaces at the center of the film. The behaviors of water and octane films with dense close-packed particle monolayers at their surfaces are very similar. In both cases, a transition from bilayer to bridging monolayer is observed at rather low capillary pressures. The implications of the above finding for particle stabilized emulsions are discussed. PMID- 15752024 TI - Synergism in the spreading of hydrocarbon-chain surfactants on polyethylene film anionic and cationic mixtures by a two-step procedure. AB - A study has been made of the adsorption, interaction, and spreading of mixtures of anionic and cationic surfactants at the aqueous solution/polyethylene (PE) interface. When a drop of an aqueous solution of an anionic or cationic hydrocarbon-chain surfactant is placed on a highly hydrophobic PE film (contact angle of water > 90 degrees ), it spreads to an area very little larger than that of a drop of water of the same volume. If the anionic and cationic hydrocarbon chain surfactant solutions are mixed prior to being applied to PE film, synergism is small, if any, and the reproducibility of the experimental results is poor. However, when the cationic and anionic aqueous solutions are applied on the PE film in a sequential manner, a remarkable synergism in spreading is observed and the results are very reproducible. The area spread by an aqueous solution of the anionic-cationic mixture may be more than 400 times that of aqueous solutions of the same volume and surfactant concentration of the individual surfactant components. Previous work in this laboratory on surfactant systems showing synergism in spreading on PE film, but only weak interaction at the aqueous solution/air interface, showed that the synergy was due to changes at the aqueous solution/PE interface and not to the changes at the aqueous solution/air or PE/air interface. Investigation of the adsorption behavior at the aqueous solution/solid interface of two of the anionic-cationic mixtures studied here indicates the reason for differences in spreading behavior observed with different anionic-cationic mixtures. The more similar the adsorption tendencies at the solid/aqueous solution interface of the anionic and cationic surfactants, and the closer their adsorption to an equimolar monolayer there, the stronger their interaction there and the greater their enhancement of the spreading. A mechanism is proposed for the synergy in spreading observed, based upon the difference between the surface tension in the precursor film at the spreading interface and that at the top of the spreading drop. PMID- 15752025 TI - Aqueous and gaseous adsorption from montmorillonite-carbon composites and from derived carbons. AB - Clay-carbon composites and the carbons derived from demineralization of the clay template were examined for their aqueous adsorption properties (2,4,6 trichlorophenol and methylene blue) and for their gas adsorption/separation abilities regarding CO(2), CH(4), and N(2) gases. The sorption results are discussed in relation with their structural properties (surface area, pore width and volume, and surface chemistry). It was found that the properties of the adsorbents depend highly on the synthetic route, for instance, on the use of clay or H(2)SO(4) as structure mediating and activating agents, respectively. Particularly, the simultaneous use of clay and H(2)SO(4) leads to a synergistic action, which imparts to the final solids the highest sorption capacity and the best potential for separation of CO(2) from gaseous mixtures of CH(4) and N(2). PMID- 15752026 TI - Interfacial reactions in confinement: kinetics and temperature dependence of the surface hydrolysis of polystyrene-block-poly(tert-butyl acrylate) thin films. AB - The effect of confinement on the kinetics of the surface hydrolysis of polystyrene-block-poly(tert-butyl acrylate) (PS(n)-b-PtBA(m)) thin films on oxidized silicon substrates in 3 M aqueous hydrochloric acid was systematically investigated. As shown by X-ray photoelectron spectroscopy (XPS) and contact angle measurements, a skin layer of acid-sensitive PtBA is present on the surface of PS(n)-b-PtBA(m) films, consistent with the lower surface tension of PtBA compared to that of PS. The thickness of the skin layer was determined by angle dependent XPS as approximately 8 nm for PS(690)-b-PtBA(1210). Tapping mode atomic force microscopy showed an increasing surface coverage of swollen poly(acrylic acid)-rich globules with increasing hydrolysis time. Using ex situ Fourier transform infrared spectroscopy, the reaction kinetics was determined quantitatively as a function of temperature, polymer film thickness, thermal pretreatment of the films, and block copolymer composition. The initial stages of the hydrolysis can be described as a pseudo-first-order reaction under all conditions investigated. The corresponding rate constants were found to be 2 orders of magnitude lower than those reported for the hydrolysis of tert-butyl acetate in solution and depended linearly on the fraction of PtBA exposed at the surface. However, the polymer film thickness, thermal pretreatment of the films, block copolymer composition, and local composition did not affect the rate constants. The negative value of the activation entropy (DeltaS(298)++ = -103 J/mol K), determined according to the Arrhenius equation and transition state theory, indicates that the tightness of the transition state is more pronounced in the PS(n)-b-PtBA(m) film compared to reactions in solution. Thus, the spatial constraints due to the incorporation of the reactive ester groups in thin polymer films are responsible for the observed reduced reactivity. PMID- 15752027 TI - Thermal study of accumulation of conformational disorders in the self-assembled monolayers of C8 and C18 alkanethiols on the Au111 surface. AB - The thermal stability of short alkanethiol CH(3)(CH(2))(7)SH (C(8)) and long C(18) self-assembled monolayers (SAMs) is investigated using grazing angle reflection-absorption infrared spectroscopy, cyclic voltammetry, and molecular dynamics simulation. We track the disordering of SAM by untilting and gauche defect accumulation with increasing temperature in the 300-440 K range, a range of interest to tribology. Molecular dynamics simulation with both fully covered and partially covered C(6), C(8), and C(18) monolayers brings out the morphological changes in the SAM, which may be associated with the observed thermal stability characteristics. The molecular dynamics simulations reveal that short-chain C(6) and C(8) alkanethiols are more defective at lower temperature than the long-chain C(18) alkanethiol. With increasing temperature disorder in the SAM, as reflected in both untilting and gauche defect accumulation, tends to saturate at temperatures below 360 K for short-chain SAMs such that any further increase in temperature, until desorption, does not lead to any significant change in conformational order. In contrast the disorder in the long-chain C(18) SAM increases monotonically with temperature beyond 360 K. Thus, in a practical range of temperature, the ability of a SAM to retain order with increasing thermal perturbations is governed by the state of disorder prior to heat treatment. This deduction derived from molecular dynamics simulation helps to rationalize the significant difference we have observed experimentally between the thermal response of short- and long-chain thiol molecules. PMID- 15752028 TI - Phase behavior and viscoelastic properties of trisilanolcyclohexyl-POSS at the air/water interface. AB - A trisilanol polyhedral oligomeric silsesquioxane (POSS), trisilanolcyclohexyl POSS (TCyP), has recently been reported to undergo a series of phase transitions from traditional Langmuir monolayers to unique rodlike hydrophobic aggregates in multilayer films that are different from "collapsed" morphologies seen in other systems at the air/water interface. This paper focuses on the phase transitions and morphology of films varying in average thickness from monolayers to trilayers and the corresponding viscoelastic properties of trisilanolcyclohexyl-POSS molecules at the air/water interface by means of surface pressure-area per molecule (Pi-A) isotherms, Brewster angle microscopy (BAM), and interfacial stress rheometry (ISR) measurements. The morphology studies by BAM reveal that the TCyP monolayer can collapse into different 3D structures by homogeneous or heterogeneous nucleation mechanisms. For homogeneous nucleation, analysis by Vollhardt et al.'s nucleation and growth model reveals that TCyP collapse is consistent with instantaneous nucleation with hemispherical edge growth at Pi = 3.7 mN.m(-1). Both surface storage (Gs') and loss (Gs") moduli obtained by ISR reveal three different non-Newtonian flow regimes that correlate with phase transitions in the Pi-A isotherms: (A) A viscous liquidlike "monolayer"; (B) a "biphasic regime"between a liquidlike viscous monolayer and a more rigid trilayer; and (C) an elastic solidlike "trilayer". These observations provide interesting insights into collapse mechanisms and structures in Langmuir films. PMID- 15752029 TI - Degradation kinetics of VX on concrete by secondary ion mass spectrometry. AB - At trace coverages on concrete surfaces, the nerve agent VX (O-ethyl S-2 diisopropylaminoethyl methyl phosphonothiolate) degrades by cleavage of the P-S and S-C bonds, as revealed by periodic secondary ion mass spectrometry (SIMS). The observed kinetics were (pseudo-) first-order, with a half-life of 2-3 h at room temperature. The rate increased with surface pH and temperature, with an apparent second-order constant of k(OH) = 0.64 M(-1) min(-1) at 25 degrees C and an activation energy of 50-60 kJ mol(-1). These values are consistent with a degradation mechanism of alkaline hydrolysis within the adventitious water film on the concrete surface. Degradation of bulk VX on concrete would proceed more slowly. PMID- 15752030 TI - Degradation mechanisms and environmental effects on perfluoropolyether, self assembled monolayers, and diamondlike carbon films. AB - The degradation mechanisms and durability of selected lubricants and environmental effects on the lubricants which could be used for microelectromechanical/nanoelectromechanical systems (MEMS/NEMS) applications were studied in this paper. The degradation of perfluoropolyether (Z-DOL), four self-assembled monolayers (SAMs)-hexadecane thiol, perfluoroalkylsilane, and alkylsilane (C8 and C18)-and diamondlike carbon (DLC) films was investigated in high vacuum. Gaseous products and friction force were detected using a quadrupole mass spectrometer and strain gauges. It is believed that triboelectrical reaction and mechanical scission cause the degradation of Z-DOL. SAMs are believed to degrade by cleavage at an interfacial bond accompanied with triboelectrical reactions. DLC is believed to degrade by mechanical shear and thermal oxidation. Environmental effects on lubricant films were studied in high vacuum, argon, and air at various humidity levels. It was found that the environment has a significant influence on the lubricant performance. The lubricant films exhibit high friction and low durability in high vacuum. Oxygen in the air can cause the thermal oxidation of SAMs and DLC films. Water molecules can act as a lubricant for Z-DOL films at a moderate humidity level, while they can penetrate the Z-DOL films at a high humidity level. Water molecules can detach the SAM molecules from the substrate, whereas, for DLC films, water molecules can act as a lubricant. PMID- 15752031 TI - Thermally- and photoinduced changes in the water wettability of low-surface-area silica and titania. AB - The surface properties of silica and titania are mainly determined by the presence, density, and type of terminal hydroxyl groups (Si-OH "silanol" and Ti OH "titanol"). Thermal treatment at elevated temperatures causes dehydroxylation on both surfaces, confirmed by streaming potential and ToF-SIMS measurements. The magnitude of the zeta potential markedly decreases after heat treatment, but the IEP is not affected. The intensity ratio MOH(+)/M(+) (M = Si or Ti), which reflects the surface density of OH groups, also decreases noticeably after high temperature treatment. The mechanism is condensation of adjacent silanol/titanol groups into siloxane/titanoxane bonds. Ultraviolet light (lambda = 254 nm) has little effect on silica but rapidly induces hydrophilicity on titania surfaces. There is a strong correlation between the amount of hydrocarbons adsorbed on the surface and the density of titanol groups (thence the water contact angle). The effect of UV radiation can be entirely attributed to photolytic decomposition of organic contaminants. Dehydroxylated titania and silica (at 1050 degrees C) show very different wetting behavior: silica is moderately hydrophobic (water contact angle of about 40 degrees), while titania is hydrophilic (0 degrees). This dissimilarity can be explained with a simple model estimating the van der Waals and acid-base interfacial interactions. PMID- 15752032 TI - Keggin ion mediated synthesis of hydrophobized Pd nanoparticles for multifunctional catalysis. AB - Development of simple and reliable protocols for the synthesis of organically soluble catalytically active metal nanoparticles is an important aspect of research in nanomaterials. We demonstrate herein the formation of Pd nanoparticles by reduction of aqueous Pd(NO(3))(2) by photoexcited Keggin ions (phosphotungstate anions). This results in the formation of Pd nanoparticles capped with with Keggin ions that render the particles negatively charged. The Keggin ion capped Pd nanoparticles may then be phase transferred into nonpolar organic solvents such as toluene by electrostatic complexation with cationic surfactants such as octadecylamine at the liquid-liquid interface. This results in a new class of catalyst wherein both the Pd core and Keggin ion shell may be used in a range of catalytic reactions leading to a truly multifunctional catalyst dispersible in organic solvents. PMID- 15752033 TI - CO2-expanded liquid deposition of ligand-stabilized nanoparticles as uniform, wide-area nanoparticle films. AB - Deposition of nanoparticles into uniform, wide-area thin films using CO(2) as an antisolvent is presented. Ligand-stabilized silver particles are controllably precipitated from organic solvents by pressurizing and expanding the solution with carbon dioxide. Subsequent addition of carbon dioxide as a dense supercritical fluid provides for removal of the organic solvent while avoiding the surface tensions common to evaporating solvents that are detrimental to nanoscale assemblies and structures. This brand new CO(2)-expanded liquid particle deposition technique allows for the targeted deposition of particles and results in more uniform and lower defect metal nanoparticle thin films than are provided by conventional solvent evaporation techniques. PMID- 15752034 TI - Influence of temperature and headgroup size on condensed-phase patterns in langmuir monolayers of some oxyethylenated nonionic surfactants. AB - The surface phase behavior in Langmuir monolayers of some oxyethylenated nonionic surfactants of the general formula C16En, with n = 1, 2, 3, and 4, at the air water interface has been studied by film balance and Brewster angle microscopy (BAM) over a wide range of temperatures. The C16E4 monolayers cannot show any indicative features of phase transition because of strong dipolar as well as hydration-induced repulsive interactions between the bulky headgroups. On the other hand, the monolayers of C16E1, C16E2, and C16E3 show a sharp cusp point followed by a pronounced plateau region in their respective isotherms with subsequent formation of a variety of structures in the two-phase coexistence region between the liquid expanded (LE) and liquid condensed (LC) phases at different temperatures. As usually observed, the domains of C16E1, which bears only one ethylene oxide (EO) unit in the headgroup, are circular at lower temperatures while fractal at higher temperatures. On the other hand, those for C16E2 and C16E3 are initially found to be irregular structures, which attain increasingly compact shape with increasing temperature, and finally become circular when the subphase temperature is 26 and 15 degrees C for C16E2 and C16E3, respectively. It is concluded that a higher degree of dehydration around the headgroup region appreciably reduces the headgroup size, which imparts to the molecules an increase in hydrophobicity, thereby a closer molecular packing. Consequently, the line tension of the interface increases, showing compact structures at higher temperatures. Since C16E1 bears only one EO unit in its headgroup, the dehydration effect cannot appreciably raise its hydrophobicity to overcome the increases in thermal motion and chain flexibility of the molecules. Rather, increases in subphase temperature result in a decrease in the line tension of the interface, giving fractal structures at higher temperatures. PMID- 15752035 TI - Surface chemical modification of waxy maize starch nanocrystals. AB - The surface of waxy maize starch nanocrystals obtained from sulfuric acid hydrolysis of native waxy maize starch granules was chemically modified using two different reagents, namely, alkenyl succinic anhydride and phenyl isocyanate. The occurrence of chemical modification was evaluated by FTIR and X-ray photoelectron spectroscopies. Contact angle measurements from which the surface energy of the materials under investigation was deduced showed that chemical modification led to more hydrophobic particles. Chemical modification altered the morphology of particles, as shown by observation by transmission electron microscopy, but not their crystallinity (X-ray diffraction analysis). PMID- 15752036 TI - Self-assembly of poly(N-isopropylacrylamide)-carrying microspheres into two dimensional colloidal arrays. AB - Droplets containing polymer particles were deposited on a substrate. Poly(N isopropylacrylamide) (PNIPAM) hydrogel and particles with PNIPAM graft chains on the surface self-assembled into a two-dimensional (2-D) superlattice when their dilute dispersions were dried on substrates. The capillary force between the particles induced ordered array formation during water evaporation. The presence of a PNIPAM layer on the particle surface gave the particles steric stability during ordered array formation. By grafting PNIPAM chains on particle surfaces by living radical polymerization, we successfully controlled the structural patterns of the colloidal arrays. These, controllable, 2-D colloidal arrays were generated on various substrates upon air-drying. PMID- 15752037 TI - Spreading of viscous liquids at high temperature: silicate glasses on molybdenum. AB - The spreading of Si-Ca-Al-Ti-O glasses on molybdenum has been investigated. By controlling the oxygen activity in the furnace, spreading can take place under reactive or nonreactive conditions. As the nucleation of the reaction product under reactive conditions is slow in comparison to the spreading kinetics, in both cases the glass front moves on the metal surface with similar spreading velocities. Spreading can be described using a molecular dynamics model where the main contribution to the wetting activation energy comes from the viscous interactions in the liquid. Enhanced interfacial diffusions in low-oxygen activities (reactive cases) form triple-line ridges that can pin the wetting front and cause a stick-slip motion. PMID- 15752038 TI - Adsorption of hydrocarbons on mesoporous SBA-15 and PHTS materials. AB - Plugged hexagonal templated silica (PHTS) materials are synthesized using a high TEOS/EO(20)PO(70)EO(20) ratio in the SBA-15 synthesis. This generates internal microporous nanocapsules or plugs in part of the channels, which could be inferred from the two-step desorption branch. These materials exhibit a tunable amount of open and plugged pores and a very high micropore volume (up to 0.24 mL/g) and are more stable than the conventional micellar templated structures known so far. In this study the adsorption properties of PHTS are investigated and compared to those of its plug-free analogue SBA-15. For this purpose nitrogen, n-hexane, n-heptane, c-hexane, 3-methylpentane, 1-hexene, and water were adsorbed on SBA-15 and PHTSs with a different ratio of open and plugged mesopores. The adsorption of n-hexane, c-hexane, n-heptane, and 3-methylpentane on SBA-15 and PHTS-A demonstrated that the presence of the plugs had an effect on the uptake of adsorbate in the low relative pressure region, the position of the capillary condensation step, and the total adsorbed amount of adsorbate. The results showed that n-heptane and 3-methylpentane cannot access part of the micropore system of SBA-15 and PHTS-A. Adsorption of c-hexane and n-hexane on PHTS-A indicated that not only the kinetic diameter but also the shape of the molecule is an important factor for being able to be adsorbed into the micropores or past the plugs. Moreover, these two adsorbates were the most efficient in filling up the available pore volume. From the adsorption of n-hexane on PHTSs with a different ratio of open and plugged pores, it was concluded that the size of the plugs differed, which depends on the synthesis conditions. Water adsorption isotherms proved SBA-15 and PHTS-B to be more hydrophobic than PHTS-A. n-Hexane, 1-hexene, and toluene were adsorbed on SBA-15 and the PHTSs to investigate the influence of the polarity of the adsorbate. The isotherms showed higher uptakes for polar adsorbates on more hydrophobic materials and vice versa. PMID- 15752039 TI - Statistical thermodynamics models for polyatomic adsorbates: application to adsorption of n-paraffins in 5A zeolite. AB - Experimental adsorption isotherms of five n-paraffins (ethane, propane, butane, pentane, and hexane) in 5A zeolite were described by means of a statistical thermodynamics model for linear adsorbates (MLA) developed by Ramirez-Pastor et al. (1999) and compared with the well-known multisite Langmuir model (MSL) of Nitta et al. (1984). The experimental data, obtained by different authors in a wide range of temperatures and pressures, were correlated by using an algorithm of multiple fitting. Two main conclusions were drawn from the analysis of experimental data: (i) for small molecules (ethane, propane), MLA is the more accurate model, validating the hypothesis of the linear rigid character of the adsorbate and reinforcing previous results obtained from the analysis of computational experiments developed for dimers and linear trimers; (ii) for large molecules (n-butane, n-pentane, n-hexane), the better performance of the MSL model suggests that the admolecules adsorb in a nonlinear structure. The isosteric heat of adsorption dependence on the number of carbons obtained from our study, ranging between 23.84 kJ/mol for ethane and 59.26 kJ/mol for hexane, showed a very good agreement with previous results reported in the literature, confirming the consistency of our analysis. PMID- 15752040 TI - Control of moisture at buried polymer/alumina interfaces through substrate surface modification. AB - Moisture absorption in poly(4-tert-butoxycarbonyloxystyrene) (PBOCSt) films supported on Al(2)O(3) sputter coated silicon wafers is measured using neutron and X-ray reflectivity. Accumulation of water at the interface during moisture exposure results in an apparent film-thickness-dependent swelling for ultrathin PBOCSt films. The swelling of a film on Al(2)O(3) is less than the swelling of a film of the same thickness on SiO(x) for films thinner than 20 nm. This is due to comparatively less moisture accumulation at the Al(2)O(3)/PBOCSt interface. A simple, zero adjustable parameter model consisting of a fixed water-rich layer at the interface and bulk swelling through the remainder of the film describes the thickness-dependent swelling quantitatively. The influence of four different Al(2)O(3) surface treatments on the moisture distribution within PBOCSt films was examined: bare Al(2)O(3), tert-butylphosphonic acid, phenylphosphonic acid, and n octyltrichlorosilane. Both the phenyl and the octyl surface treatments reduce the accumulation of water at the polymer/substrate interface. The tert-butyl treatment does not reduce the interfacial water concentration, presumably due to insufficient surface coverage. PMID- 15752041 TI - Use of block copolymer-stabilized cadmium sulfide quantum dots as novel tracers for laser scanning confocal fluorescence imaging of blend morphology in polystyrene/poly(methyl methacrylate) films. AB - This paper describes the first use of polymer-coated quantum dots (QDs) as fluorescent tracers for LSCFM imaging of phase morphology in polymer blends. Cadmium sulfide (CdS) QDs stabilized at the surface with a PS-b-PAA block copolymer are shown to be well dispersed via their polystyrene (PS) brush layer in the PS phase of solvent-cast 40/60 (w/w) PS/PMMA blends. The QDs are excluded from the PMMA phase, providing excellent fluorescence contrast for LSCFM imaging of the phase-separated blends. The presence of PS-b-PAA-stabilized QDs does not appear to affect the blend morphology, since the observed morphologies are the same when the percentage of QDs within the PS phase is varied from 10 to 50 wt %. These QD fluorescent tracers are used to characterize several aspects of blend morphology in solvent-cast 40/60 PS/PMMA blends containing PS homopolymer with either 100 (low molecular weight) or 1250 (high molecular weight) repeat units. In the PS(1250)/PMMA blends, a percolating distribution of PMMA droplets (2-25 mum) in a PS matrix is observed in the bulk, and a distinct inversion in the continuous phase is found near the glass substrate. In the PS(100)/PMMA blends, a "phase-in-phase" morphology is found, consisting of large PS domains (20-100 mum) dispersed in a PMMA continuous phase and small PMMA domains (1-2 mum) scattered throughout the larger PS droplets. The observed change in blend structure is attributed to a lower interfacial tension for the lower molecular weight PS. PMID- 15752042 TI - Effect of mono-CDNP substitution of lysine residues on the redox reaction of cytochrome c electrostatically adsorbed on a mercaptoheptanoic acid modified Au(111) surface. AB - The effect of charge-inverting modification of single surface lysine residue on the electron transfer (ET) reaction of horse heart cytochrome c (cyt c) is examined for 12 different types of mono-4-chloro-2,5-dinitrobenzoic acid substituted cyt c (mCDNPc) adsorbed on a Au(111) electrode modified with a self assembled monolayer (SAM) of 7-mercapto-heptanoic acid (MHA). A negative shift in the redox potential by 10-35 mV as compared to that of native cyt c and a monolayer coverage in the range of 13-17 pmol cm(-2) are observed for electroactive mCDNPc's. The magnitude of the decrease in the ET rate constant (k(et)) of mCDNPc's compared with that of native cyt c depends on the position of the CDNP substitution. For mCDNPc's in which the modified lysine residue is outside of the interaction domain of cyt c with the SAM, the ratio of the k(et) of mCDNPc to that of native cyt c is correlated to the change in the dipole moment vector of cyt c due to the CDNP modification. This correlation suggests that the dipole moment of cyt c determines its orientation of adsorption on the SAM of MHA and significantly affects the rate of the ET. The CDNP modification of lysine residues at the interaction domain significantly decreases the rate, demonstrating the importance of the local charge environment in determining the rate of ET. PMID- 15752043 TI - Alpha-helix-inducing dimerization of synthetic polypeptide scaffolds on gold. AB - Designed, synthetic polypeptides that assemble into four-helix bundles upon dimerization in solution were studied with respect to folding on planar gold surfaces. A model system with controllable dimerization properties was employed, consisting of negatively and positively charged peptides. Circular dichroism spectroscopy and surface plasmon resonance based measurements showed that at neutral pH, the peptides were able to form heterodimers in solution, but unfavorable electrostatic interactions prevented the formation of homodimers. The dimerization propensity was found to be both pH- and buffer-dependent. A series of infrared absorption-reflection spectroscopy experiments of the polypeptides attached to planar gold surfaces revealed that if the negatively charged peptide was immobilized from a loading solution where it was folded, its structure was retained on the surface provided it had a cysteine residue available for anchoring to gold. If it was immobilized as random coil, it remained unstructured on the surface but was able to fold through heterodimerization if subsequently exposed to a positively charged polypeptide. When the positively charged peptide was immobilized as random coil, heterodimerization could not be induced, probably because of high-affinity interactions between the charged primary amine groups and the gold surface. These observations are intended to pave the way for future engineering of functional surfaces based on polypeptide scaffolds where folding is known to be crucial for function. PMID- 15752044 TI - Sequence-specific binding of DNA to liposomes containing di-alkyl peptide nucleic acid (PNA) amphiphiles. AB - We present a method to covalently attach peptide nucleic acid (PNA) to liposomes by conjugation of PNA peptide to charged amino acids and synthetic di-alkyl lipids ("PNA amphiphile," PNAA) followed by co-extrusion with disteroylphosphatidylcholine (DSPC) and cholesterol. Attachment of four Glu residues and two ethylene oxide spacers to the PNAA was required to confer proper hydration for extrusion and presentation for DNA hybridization. The extent of DNA oligomer binding to 10-mer PNAA liposomes was assessed using capillary zone electrophoresis. Nearly all PNAs on the liposome surface are complexed with a stoichiometric amount of complementary DNA 10-mers after 3-h incubation in pH 8.0 Tris buffer. No binding to PNAA liposomes was observed using DNA 10-mers with a single mismatch. Longer DNA showed a greatly attenuated binding efficiency, likely because of electrostatic repulsion between the PNAA liposome double layer and the DNA backbone. Langmuir isotherms of PNAA:DSPC:chol monolayers indicate miscibility of these components at the compositions used for liposome preparation. PNAA liposomes preserve the high sequence-selectivity of PNAs and emerge as a useful sequence tag for highly sensitive bioanalytical devices. PMID- 15752045 TI - Oligo(ethylene glycol) containing polymer brushes as bioselective surfaces. AB - The nitroxide-mediated polymerization of styrenic monomers containing oligo(ethylene glycol) (OEGn) moieties was chosen for the preparation of biocompatible polymer brushes tethered to silicon oxide surfaces due to the broad range of monomer structures available and the use of a nonmetallic initiator. These surfaces were characterized by near-edge X-ray absorption fine structure and water contact angle measurements. The biocompatibility of these grown polymer brushes was studied and compared with deposited assemblies of surface-bound OEGn terminated silanes with selected chain lengths. Grown polymer brushes with short OEGn side chains suppressed protein adsorption significantly more than the deposited assemblies of short OEGn chains, and this was attributed to higher surface coverage by the brushes. Cell adhesion studies confirmed that OEGn containing polymer brushes are particularly effective in preventing nonspecific adhesion. Studies of protein adsorption and cell localization carried out with specific ligands on surfaces patterned demonstrated the potential of these surface-tethered polymer brushes for the formation of micro- and nanoscale devices. PMID- 15752046 TI - Nanoscale properties of mixed fengycin/ceramide monolayers explored using atomic force microscopy. AB - To gain insight into the interactions between fengycin and skin membrane lipids, mixed fengycin/ceramide monolayers were investigated using atomic force microscopy (AFM) (monolayers supported on mica) and surface pressure-area isotherms (monolayers at the air-water interface). AFM topographic images revealed phase separation in mixed monolayers prepared at 20 degrees C/pH 2 and composed of 0.25 and 0.5 fengycin molar ratios, in the form of two-dimensional (2 D) hexagonal crystalline domains of ceramide surrounded by a fengycin-enriched fluid phase. Surface pressure-area isotherms as well as friction and adhesion AFM images confirmed that the two phases had different molecular orientations: while ceramide formed a highly ordered phase with crystalline chain packing, fengycin exhibited a disordered fluid phase with the peptide ring lying horizontally on the substrate. Increasing the temperature and pH to values corresponding to the skin parameters, i.e., 37 degrees C/pH 5, was found to dramatically affect the film organization. At low fengycin molar ratio (0.25), the hexagonal ceramide domains transformed into round domains, while at higher ratio (0.5) these were shown to melt into a continuous fengycin/ceramide fluid phase. These observations were directly supported by the thermodynamic analysis (deviation from the additivity rule, excess of free energy) of the monolayer properties at the air water interface. Accordingly, this study demonstrates that both the environmental conditions (temperature, pH) and fengycin concentration influence the molecular organization of mixed fengycin/ceramide monolayers. We believe that the ability to modulate the formation of 2-D domains in the skin membrane may be an important biological function of fengycin, which should be increasingly investigated in future pharmacological research. PMID- 15752047 TI - Modeling the stability and the motion of DNA nucleobases on the gold surface. AB - We simulate the structure and dynamics of the four DNA bases on the most stable gold surface. The experimental adsorption energies are reproduced to about 1 kcal mol(-1), and the existence of anchor points in the molecules is evidenced. The simulations also show that the bases drift on the gold surface with a degree of mobility that is not inversely proportional to the experimental (and calculated) desorption energies. When the same type of calculations is applied to pairs of bases it is seen that for at least two of them, namely GG and TT, there is a cooperative effect that increases their adsorption energy with respect to those of the single molecules. The molecular mobility on the surface is still present when a pair of interacting bases is considered. PMID- 15752048 TI - Combining optical lithography with rapid microwave heating for the selective growth of Au/Ag bimetallic core/shell structures on patterned silicon wafers. AB - We demonstrate a novel approach for the production of patterned films of nanometer-sized Au/Ag bimetallic core/shell nanoparticles (NPs) on silicon wafers. In this approach, we first self-assembled monodisperse Au NPs, through specific Au...NH(2) interactions, onto a silicon substrate whose surface had been modified with a pattern of 3-aminopropyltrimethoxysilane (APTMS) groups to form a sandwich structure having the form Au NPs/APTMS/SiO(2). These Au NPs then served as seeds for growing the Au/Ag bimetallic core/shell NPs: we reduced silver ions to Ag metal on the surface of Au seeds under rapid microwave heating in the presence of sodium citrate. Energy-dispersive X-ray analysis confirmed that the Au/Ag bimetallic core/shell NPs grew selectively on the regions of the surface of the silicon wafer that had been patterned with the Au seeds. Scanning electron microscopy images revealed that we could synthesize well-scattered, high-density (>82%) thin films of Au/Ag bimetallic core/shell NPs through the use of this novel strategy. The patterned structures that can be formed are simple to produce, easily controllable, and highly reproducible; we believe that this approach will be useful for further studies of nanodevices and their properties. PMID- 15752049 TI - Synthesis, characterization of dihydrolipoic acid capped gold nanoparticles, and functionalization by the electroluminescent luminol. AB - The use of gold nanoparticles as biological probes requires the improvement of colloidal stability. Dihydrolipoic acid (DHLA), a dithiol obtained by the reduction of thioctic acid, appears therefore very attractive for the stabilization and the further functionalization of gold nanoparticles because DHLA is characterized by a carboxylic acid group and two thiol functions. The ionizable carboxylic acid groups ensure, for pH > or = 8, the water solubility of DHLA-capped gold (Au@DHLA) nanoparticles, prepared by the Brust protocol, and the stability of the resulting colloid by electrostatic repulsions. Moreover almost all DHLA, adsorbed onto gold, adopts a conformation allowing their immobilization by both sulfur ends. It is proved by sulfur K-edge X-ray absorption near edge structure spectroscopy, which appears as an appropriate tool for determining the chemical form of sulfur atoms present in the organic monolayer. Such a grafting renders the DHLA monolayers more resistant to displacement by dithiothreitol than mercaptoundecanoic acid monolayers. The presence of DHLA on gold particles allows their functionalization by the electroluminescent luminol through amine coupling reactions assisted by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. As a luminol-functionalized particle is nine times as bright as a single luminol molecule, the use of the particles as a biological probe with a lower threshold of detection is envisaged. PMID- 15752050 TI - On the formation of narrowly polydispersed PMMA by surface initiated polymerization (SIP) from AIBN-coated/intercalated clay nanoparticle platelets. AB - Various free radical surface initiated polymerization (SIP) conditions were investigated on clay nanoparticles coated with monocationic 2,2' azobisisobutyronitrile (AIBN) type free radical initiators. Interesting differences in the mechanism of polymer nanocomposite product formation and the role of nanoparticle surface bound AIBN initiators were observed on three types of poly(methyl methacrylate) (PMMA) polymerization conditions: bulk, suspension, and solution. X-ray diffraction (XRD) and differential scanning calorimetry (DSC) measurements confirmed the attachment of the initiator on the clay particles without decomposition of the azo group. XRD and transmission electron microscopy (TEM) showed that a well-dispersed structure was accomplished only by bulk and solution SIP. The suspension SIP product consisted of a partially exfoliated structure as shown by XRD and clay particle aggregate formation as shown by TEM. In general, the molecular weights (MWs) of the surface bound polymers were found to be lower than those of the free polymer. Using the same clay loading and initiator concentrations, we observed that relatively higher MW polymers were obtained from suspension and bulk polymerizations in contrast to solution method. However, the most interesting observation is that the surface bound polymers (on all three conditions) showed much narrower polydispersity compared to that of a typical AIBN type free radical polymerization. PMID- 15752051 TI - Synthesis of nanoporous graphite-derived carbon-silica composites by a mechanochemical intercalation approach. AB - A mechanochemical intercalation approach which applies a simple mechanical milling to induce intercalation reaction was applied to introduce controlled amount of tetraethoxylsilane (TEOS) into surfactant-preexpanded graphite oxide, and the relationships between the intercalation structure, the porosities of the calcined products, and the Si addition were examined. It was found that a small added amount of TEOS produced a more expanded ordered layer structure with the interlayer distance and silicon content increasing with the amount of TEOS added, although a large amount of added TEOS easily induces layer delamination, resulting in a less ordered structure. The silica structure in the composite is changed from a disordered structure having enhanced bond strain to a condensed silica network when the amount of TEOS added increases. The porosities of the final calcined samples increase with the increase of silicon content but then decrease slightly after reaching a maximum where silicon content starts to become constant, indicating that both silicon content and the composition state of silica particles and carbon layers play important roles in porosity formation. PMID- 15752052 TI - Photocatalytic activity of a hierarchically macro/mesoporous titania. AB - Light-harvesting macroporous channels have been successfully incorporated into a mesoporous TiO(2) framework to increase its photocatalytic activity. This bimodal porous material was characterized by X-ray diffractometry in both low-angle and wide-angle ranges, N(2) adsorption-desorption analysis, scanning and transmission electron microscopy, FT-IR, and diffuse reflectance spectroscopy. Ethylene photodegradation in gas-phase medium was employed as a probe reaction to evaluate the photocatalytic reactivity of the catalysts. The results reveal that sintering temperature significantly affects the structural stability and photocatalytic activity of titania. The catalyst which calcined at 350 degrees C possessed an intact macro/mesoporous structure and showed photocatalytic reactivity about 60% higher than that of commercial P25 titania. When the sample was calcined at 500 degrees C, the macroporous structure was retained but the mesoporous structure was partly destroyed. Further heating at temperatures above 600 degrees C destroyed both macro- and mesoporous structures, accompanied by a loss in photocatalytic activity. The high photocatalytic performance of the intact macro/mesoporous TiO(2) may be explained by the existence of macrochannels that increase photoabsorption efficiency and allow efficient diffusion of gaseous molecules. PMID- 15752054 TI - Cubic phase nanoparticles (Cubosome): principles for controlling size, structure, and stability. AB - Methods and compositions for producing lipid-based cubic phase nanoparticles were first discovered in the 1990s. Since then a number of studies have been presented, but little is known about how to control key properties such as particle size, morphology, and stability of cubic phase dispersions. In the present work we give examples of how these properties can be tuned by composition and processing conditions. Importantly we show that stable particle dispersions with consistent size and structure can be produced by a simple processing scheme comprising a homogenization and heat treatment step. PMID- 15752053 TI - Electrostatics of PEGylated micelles and liposomes containing charged and neutral lipopolymers. AB - The electrostatics of large unilamellar vesicles (LUVs) of various lipid compositions were determined and correlated with steric stabilization. The compositional variables studied include (a) degree of saturation, comparing the unsaturated egg phosphatidylcholine (EPC) and the fully hydrogenated soy phosphatidylcholine (HSPC) as liposome-forming lipids; (b) the effect of 40 mol % cholesterol; (c) the effect of mole % of three methyl poly(ethylene glycol) (mPEG)-lipids (the negatively charged mPEG-distearoyl phosphoethanolamine (DSPE) and two uncharged lipopolymers, mPEG-distearoyl glycerol (DSG) and mPEG oxycarbonyl-3-amino-1,2-propanediol distearoyl ester (DS)); and (d) the negatively charged phosphatidyl glycerol (PG). The lipid phases were as follows: liquid disordered (LD) for the EPC-containing LUV, solid ordered (SO) for the HSPC-containing LUV, and liquid ordered (LO) for either of those LUV with the addition of 40 mol % cholesterol. The LUV zeta potential and electrical surface potential (psi(0)) were determined. It was found that progressive addition of mPEG(2k)-DSPE to liposomes increases negative surface potential and reduces surface pH to a similar extent as the addition of PG. However, due to the "hidden charge effect", zeta potential was more negative for liposomes containing PG than for those containing mPEG(2k)-DSPE. Replacing mPEG-DSPE with mPEG(2k)-DS or mPEG DSG had no effect on surface pH and surface potential, and zeta potential was approximately zero. Addition of low concentrations of cationic peptides (protamine sulfate and melittin) to PG- or mPEG-DSPE-containing liposomes neutralized the liposome negative surface potential to a similar extent. However, only in liposomes containing PG, did liposome aggregation occur. Replacing the negatively charged lipopolymer mPEG-DSPE with the neutral lipopolymers mPEG-DS or mPEG-DSG eliminates or reduces such interactions. The relevance of this effect to the liposome performance in vivo is discussed. PMID- 15752055 TI - Photochemical preparation of poly(N-vinyl-2-pyrrolidone)-stabilized platinum colloids and their deposition on titanium dioxide. AB - Preparation processes for Pt-deposited TiO(2) (Pt/TiO(2)) by the synthesis of Pt nanoparticles and their deposition were pursued by transmission electron microscopy, extended X-ray absorption fine structure, UV-vis spectroscopy, and Fourier transform infrared spectroscopic studies. Colloidal dispersions of Pt particles stabilized by poly(N-vinyl-2-pyrrolidone) (PVP) were photochemically synthesized in aqueous ethanol solution. The average diameter of Pt particles was estimated to be 2.0 +/- 0.5 nm, which was almost unchanged by changing the reducing agent from ethanol to methanol and 2-propanol. The PVP-stabilized Pt particles were distributed over a TiO(2) surface only by mixing the Pt colloidal dispersions and TiO(2). CO was chemically coordinated on the Pt particles on a TiO(2) surface after heat treatment was carried out in an O(2) flow at 673 K to completely remove the residual PVP on Pt/TiO(2). Hydrogen reduction at 473 K did not increase the amount of CO adsorbed on Pt sites. The Pt/TiO(2) catalyst after the oxidation treatment showed higher activity for CO photooxidation than that obtained for pure TiO(2) catalyst. The CO photooxidation rate was not unchanged by the H(2) reduction. PMID- 15752056 TI - Infrared study of the interaction of charged silica particles with TiO2 particles containing adsorbed cationic and anionic polyelectrolytes. AB - Attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy was used to study the adsorption of charged silica particles onto TiO(2) particles coated with anionic sodium polyacrylate (NaPA) or cationic poly(diallyldimethylammonium) chloride (PDADMAC). To the best of our knowledge, this is the first time that IR spectroscopy has been used to study the interaction of a polymer layer on one particle with a second different particle. The results show that, once adsorbed on the TiO(2) particle, the PDADMAC or the NaPA does not transfer to the silica particles. In the case of NaPA coated TiO(2), positively charged silica particles deposit on the TiO(2) and this is accompanied by a change in the relative intensities of the bands due to COOH and COO(-) groups. From this change in band intensity, it is calculated that only approximately 6% of the COO(-) groups located in the loops and tails bind to the silica particle. This shows that the polymer bridges the two particles through an electrostatic interaction with the outer COO(-) groups. Similarly, in the case of the TiO(2) particles coated with PDADMAC, negatively charged silica deposits on the TiO(2) and this is accompanied by an increase in intensity of the symmetric bending mode of the (+)N(CH(3))(3) group. This change in band intensity arises from the binding of these cationic sites of the polymer to the negative surface sites on the silica. PMID- 15752057 TI - Solvent-induced textural changes of as-synthesized mesoporous alumina, as reported by spin probe electron spin resonance spectroscopy. AB - The effect of solvent used during the synthesis and postsynthesis treatment on textural properties of organized mesoporous aluminas was investigated and related to the behavior of spin probes studied by electron spin resonance (ESR) spectroscopy. It was found that the structure of surfactant aggregates serving in the as-synthesized precipitates as templates could be easily modified by treatment with different solvents. This treatment induces corresponding variations in surface areas, mesopore volumes, and mesopore diameters of the final products. The ESR spectrum of 5-doxyl stearic acid spin probe properly reflects the changes in template structure based on changes of the solvent used and represents an early indicator of the corresponding textural modifications of the mesoporous alumina. PMID- 15752058 TI - Internal structure of polyelectrolyte multilayers probed via neutral impact collision ion scattering spectroscopy. AB - Layering in polyelectrolyte multilayer films has been studied by neutral impact collision ion scattering spectroscopy. The method affords a direct look at vertical ordering within these films at the nanometer scale. By labeling certain polyelectrolyte layers with heavy atom (Ru) probes, sufficient contrast has been obtained to visualize and quantify the distribution of these labeled polyelectrolytes throughout the film. The results indicate that the materials under investigation here produce linear film growth with very limited layer interpenetration. The interdigitation length between neighboring layers within the film is measured as 3.9 nm, which is approximately 1.4 times the thickness of an individual polycation/polyanion pair and is slightly less than the measured air/film roughness (4.7 nm). Detailed analysis shows that under the conditions used and at the depths probed in this study, the observed layer thickness is not significantly broadened by either instrumental or stochastic factors. PMID- 15752059 TI - Amphiphilic hydrogel nanoparticles. Preparation, characterization, and preliminary assessment as new colloidal drug carriers. AB - Inverse emulsion photopolymerization of acrylated poly(ethylene glycol)-bl poly(propylene glycol)-bl-poly(ethylene glycol) and poly(ethylene glycol) was successfully employed to prepare stable, cross-linked, amphiphilic nanoparticles. Even at low emulsifier concentrations (2%) and high water-to-hexane weight ratios (35/65), the stability of the inverse emulsion allowed for the formation of well defined colloidal material. Inverse emulsion characteristics and polymerization conditions could be controlled to vary the size of the nanoparticles between 50 and 500 nm. The presence of hydrophobic nanodomains within these otherwise hydrophilic nanoparticles was verified by using pyrene as a microenvironmentally sensitive probe. The hydrophobic poly(propylene glycol)-rich domains appear to be suitable for incorporation of hydrophobic drugs, encapsulating Doxorubicin up to 9.8% (w/w). We believe that the complex nano-architecture of these materials makes them a potentially interesting colloidal drug delivery carrier system and that the method should be useful for a number of amphiphilic macromolecular precursors. PMID- 15752060 TI - Effects of film-forming conditions on surface properties and structures of diblock copolymer with perfluoroalkyl side chains. AB - Film-forming conditions (cast solvents and film-forming technique: casting or spin-coating) are found to regulate the surface free energy of a diblock copolymer composed of methyl methacrylate and 2-perfluorooctylethyl methacrylate (PMMA-b-PFEMA). The surface properties and structures both in the solid state and in the solution of this polymer were investigated using dynamic contact angle, X ray photoelectron spectroscopy, X-ray diffraction, reflection-adsorption Fourier transform infrared spectroscopy, dynamic light scattering, and surface pressure measurements. The surface free energy of the PMMA-b-PFEMA film drastically changed from 7.8 to 28.4 mJ/m(2), depending on the film-forming conditions. For the film cast from the micellar solution, the surface free energy is governed by the unimers, localized at the air-solution interface. On the other hand, higher amounts of micelles with the laid perfluoroalkyl side chains were exposed on the surface of the spin-coated film, which yielded a relatively high surface free energy. These findings demonstrated a significant effect of the in-solution molecular association on the surface structures and properties of the solid. In particular, the in-solution unimers played the most important role in lowering the surface free energy of the PMMA-b-PFEMA films. PMID- 15752061 TI - Hofmeister effects in surface tension of aqueous electrolyte solution. AB - The surface tension of electrolyte solutions shows marked specific ion effects. We here show an important role for both ionic solvation energies and ionic dispersion potentials in determining this ion specific surface tension of salt solutions. The ion self-free energy changes when an ion moves from bulk solution into the interfacial region, with its decreasing water density profile. We will show that the solvation energies of different ions correlate very well with the surface tension of salt solutions. Inclusion of this distance-dependent self-free energy contribution brings qualitative agreement with experiments and the right Hofmeister series. This is so not only for surface tension changes but also for measured surface potentials. The inclusion of ionic dispersion interaction potentials further improves the agreement with experiments. We discuss how further progress in the theory of the surface tension of salts can be achieved. PMID- 15752062 TI - Modeling droplets on superhydrophobic surfaces: equilibrium states and transitions. AB - We present a lattice Boltzmann solution of the equations of motion describing the spreading of droplets on topologically patterned substrates. We apply it to model superhydrophobic behavior on surfaces covered by an array of micrometer-scale posts. We find that the patterning results in a substantial increase in contact angle, from 110 degrees to 156 degrees. The dynamics of the transition from drops suspended on top of the posts to drops collapsed in the grooves is described. PMID- 15752063 TI - Comment on tilt of atomic force microscope cantilevers: effect on spring constant and adhesion measurements. PMID- 15752067 TI - Different contribution of interleukin-6 and cortisol activity to total plasma fibrin concentration and to acute mental stress-induced fibrin formation. AB - Acute mental stress may contribute to atherosclerosis by affecting inflammation and coagulation; however, the crosstalk between inflammation and coagulation during stress has not been studied. In the present study, we investigated the association of plasma fibrinogen, plasma IL-6 (interleukin-6) and free salivary cortisol with the procoagulant marker D-dimer reflecting fibrin formation both over a 2-h period and in response to acute mental stress. Twenty-one male volunteers (mean age, 47+/-8 years) underwent the Trier Social Stress Test combining a 3-min preparation phase, a 5-min job interview and 5-min mental arithmetic test before an audience. IL-6, fibrinogen, D-dimer and cortisol were measured immediately before and after stress, and after 45 min and 105 min of recovery from stress. Two distinct areas under the curve were computed to obtain integrated measures of total protein activity over the entire 2-h period and of stress reactivity of proteins. IL-6 (P < 0.001), fibrinogen (P = 0.001), D-dimer (P = 0.021) and cortisol (P < 0.001) had all significantly changed across the four time points assessed, as determined by ANOVA. For the entire 2-h period, total fibrinogen activity (R2 = 0.33, P = 0.007) and total cortisol activity (DeltaR2 = 0.17, P = 0.034) explained 50% of the variance in total D-dimer activity. Stress-induced changes in fibrinogen (R2 = 0.47, P = 0.001) and IL-6 (DeltaR2 = 0.18, P = 0.008) together explained 65% of the variance in D-dimer reactivity to stress. Total fibrin formation was independently predicted by fibrinogen and hypothalamo-pituitary-adrenal activity. Pro-inflammatory and procoagulant changes with stress were associated. Aside from fibrinogen reactivity, IL-6 reactivity was an independent predictor of stress-induced fibrin formation. PMID- 15752064 TI - Basal immunoglobulin signaling actively maintains developmental stage in immature B cells. AB - In developing B lymphocytes, a successful V(D)J heavy chain (HC) immunoglobulin (Ig) rearrangement establishes HC allelic exclusion and signals pro-B cells to advance in development to the pre-B stage. A subsequent functional light chain (LC) rearrangement then results in the surface expression of IgM at the immature B cell stage. Here we show that interruption of basal IgM signaling in immature B cells, either by the inducible deletion of surface Ig via Cre-mediated excision or by incubating cells with the tyrosine kinase inhibitor herbimycin A or the phosphatidylinositol 3-kinase inhibitor wortmannin, led to a striking "back differentiation" of cells to an earlier stage in B cell development, characterized by the expression of pro-B cell genes. Cells undergoing this reversal in development also showed evidence of new LC gene rearrangements, suggesting an important role for basal Ig signaling in the maintenance of LC allelic exclusion. These studies identify a previously unappreciated level of plasticity in the B cell developmental program, and have important implications for our understanding of central tolerance mechanisms. PMID- 15752069 TI - Towards a better understanding of the substrate specificity of the UDP-N acetylglucosamine C4 epimerase WbpP. AB - WbpP is the only genuine UDP-GlcNAc (UDP-N-acetylglucosamine) C4 epimerase for which both biochemical and structural data are available. This represents a golden opportunity to elucidate the molecular basis for its specificity for N acetylated substrates. Based on the comparison of the substrate binding site of WbpP with that of other C4 epimerases that convert preferentially non-acetylated substrates, or that are able to convert both acetylated and non-acetylated substrates equally well, specific residues of WbpP were mutated, and the substrate specificity of the mutants was determined by direct biochemical assays and kinetic analyses. Most of the mutations tested were anticipated to trigger a significant switch in substrate specificity, mostly towards a preference for non acetylated substrates. However, only one of the mutations (A209H) had the expected effect, and most others resulted in enhanced specificity of WbpP for N acetylated substrates (Q201E, G102K, Q201E/G102K, A209N and S143A). One mutation (S144K) totally abolished enzyme activity. These data indicate that, although all residues targeted in the present study turned out to be important for catalysis, determinants of substrate specificity are not confined to the substrate-binding pocket and that longer range interactions are essential in allowing proper positioning of various ligands in the binding pocket. Hence prediction or engineering of substrate specificity solely based on sequence analysis, or even on modelling of the binding pocket, might lead to incorrect functional assignments. PMID- 15752068 TI - C/EBPalpha activates the transcription of triacylglycerol hydrolase in 3T3-L1 adipocytes. AB - TGH (triacylglycerol hydrolase) catalyses the lipolysis of intracellular stored triacylglycerol. To explore the mechanisms that regulate TGH expression in adipose tissue, we studied the expression of TGH during the differentiation of 3T3-L1 adipocytes. TGH mRNA and protein levels increased dramatically in 3T3-L1 adipocytes compared with pre-adipocytes. Electrophoretic mobility shift assays demonstrated enhanced binding of nuclear proteins of adipocytes to the distal murine TGH promoter region (-542/-371 bp), yielding one adipocyte-specific migrating complex. Competitive and supershift assays demonstrated that the distal TGH promoter fragment bound C/EBPalpha (CCAAT/enhancer-binding protein alpha). Transient transfections of different mutant TGH promoter-luciferase constructs into 3T3-L1 adipocytes and competitive electromobility shift assays showed that the C/EBP-binding elements at positions -470/-459 bp and -404/-390 bp are important for transcriptional activation. Co-transfection with C/EBPalpha cDNA and TGH promoter constructs in 3T3-L1 pre-adipocytes demonstrated that C/EBPalpha increased TGH promoter activity. Ectopic expression of C/EBPalpha in NIH 3T3 cells activated TGH mRNA expression without causing differentiation into adipocytes. These experiments directly link increased TGH expression in adipocytes to transcriptional regulation by C/EBPalpha. This is the first evidence that C/EBPalpha participates directly in the regulation of an enzyme associated with lipolysis. PMID- 15752070 TI - PathoPlant: a database on plant-pathogen interactions. AB - Pathogen recognition and signal transduction during plant pathogenesis is essential for the activation of plant defense mechanisms. To facilitate easy access to published data and to permit comparative studies of different pathogen response pathways, a database is indispensable to give a broad overview of the components and reactions so far known. PathoPlant has been developed as a relational database to display relevant components and reactions involved in signal transduction related to plant-pathogen interactions. On the organism level, the tables 'plant', 'pathogen' and 'interaction' are used to describe incompatible interactions between plants and pathogens or diseases. On the molecular level, plant pathogenesis related information is organized in PathoPlant's main tables 'molecule', 'reaction' and 'location'. Signal transduction pathways are modeled as consecutive sequences of known molecules and corresponding reactions. PathoPlant entries are linked to associated internal records as well as to entries in external databases such as SWISS-PROT, GenBank, PubMed, and TRANSFAC. PathoPlant is available as a web-based service at http://www.pathoplant.de. PMID- 15752071 TI - An organizational model of transcription factor binding sites for a histone promoter in D. melanogaster. AB - The Drosophila H2A-H2B histone spacer, a small region that functions as a bidirectional promoter for the gene pair, was used as a test sequence for generation of a computationally derived organizational model of transcription factor (TF) binding sites. Expression studies of the spacer revealed that it contains the necessary sequences to confer replication-dependent transcription in partially synchronized cells in culture. Informatics analysis of the spacer uncovered a number of binding sites for specific TFs, none of which had been previously associated with this particular promoter. Each of the TFs in the promoter organizational model are also known to participate in stages of fly development that are characterized by DNA replication and/or cell division, thus providing a biologically functional rationale for an association. Moreover, phylogenetic analysis of the binding sites provides evidence for evolutionary conservation of the essential features of the organizational model. The model, if correct, provides information about the molecules that couple developmental specific demands and histone gene transcription. PMID- 15752072 TI - Consensus based validation of membrane porins. AB - Porins are outermembrane beta-barrel proteins. They have varied biological functionality ranging from phage receptors, immunogenicity, pathogenicity to apoptosis. However, only a small number has been structurally and functionally characterised. A validation mechanism and a database of porins would be useful for target selection in proteomics and structural genomics work. Here we report a validation mechanism developed for membrane porins. A database server for porins, PRNDS, has been created containing experimentally proven porins and likely putative porins. Each porin is validated and ranked using a weighted scoring system developed based on six, structure and sequence based criteria. The server also predicts possible porins. PMID- 15752074 TI - Clusters of proteins in archaeal and bacterial proteomes using compositional analysis. AB - In silico proteomics complements computational genomics in characterizing genome evolution. Here we examine cluster patterns in archaeal and bacterial proteomes using compositional properties of protein sequences in contrast to the traditionally used sequence alignment procedures. Application of standard Principal Component Analysis to the multi-dimensional data identified cluster patterns. Two types of cluster patterns exist in bacterial proteomes. Proteomes of type I have one major cluster with few isolated points in space revealing an underlying largely homogeneous compositional structure. In type II proteomes two clusters of protein distribution were discernible. The two clusters differ in size and were separated from each other although the boundary was somewhat fuzzy. Proteins falling in the major cluster were labeled as 'typical' and proteins of the minor cluster were called 'atypical'. The atypical proteins were mapped to Cluster of Orthologous Groups. Species distribution in COGs maps with respect to atypical proteins illuminated the biological relationships of extreme diversity among the archaeal members and of diversity among bacteria in relation to their niche. Amino acids that were over-represented in the atypical proteins had higher biosynthetic cost compared to 'typical' ribosomal proteins. However, archaea and bacteria economize by preferring the less costly amino acid to others closely related in chemical structure. Further, over-representation of serine in atypical proteins of archaeal members suggests re-examining these proteomes for the presence of Serine/Threonine phosphatases and kinases in Archaea. Our computational procedure can serve as a useful addition to the existing tools for carrying out in silico proteomics. PMID- 15752073 TI - Identification and analysis of a new family of bacterial serine proteinases. AB - A family of hypothetical proteins, identified predominantly from archaeal genomes, has been analyzed in order to understand its functional characteristics. Using extensive sequence similarity searches it is inferred that this family is remotely related (best sequence identity is 19%) to ClpP proteinases that belongs to serine proteinase class. This family of hypothetical proteins is referred to as SDH proteinase family based on conserved sequential order of Ser, Asp and His residues and predicted serine proteinase activity. Results of fold recognition of SDH family sequences confirmed the remote relationship between SDH proteinases and Clp proteinases and revealed similar tertiary location of putative catalytic triad residues critical for serine proteinase function. However, the best sequence alignment we could obtain suggests that while catalytic Ser is conserved across Clp and SDH proteinases the location of the other catalytic triad residues, namely, His and Asp are swapped in their amino acid alignment positions and hence in 3-D structure. The evidence of conserved catalytic triad suggests that SDH could be a new family of serine proteinases with the fold of Clp proteinase, however sharing the catalytic triad order of carboxypeptidase clan. Signal peptide sequence identified at the N-terminus of some of the homologues suggests that these might be secretory serine proteinases involved in cleavage of extracellular proteins while the remote homologues, ClpP proteinases, are known to work in intracellular environment. PMID- 15752075 TI - STR2: a structure to string approach for locating G-box riboswitch shapes in pre selected genes. AB - Traditional sequence-based search methods such as BLAST and FASTA can be used to identify sequence similarities. Recently, there is a growing interest in performing RNA shape similarity searches inside selected genes to locate RNA structure motifs that are known to possess functionally important roles. For example, in the newly discovered RNA genetic control elements called "riboswitches", the box domain is known to be highly conserved among various bacterial species in both its nucleotide composition and shape. However, in non bacterial species, shape conservation is likely to become more important than sequence conservation when searching for riboswitch patterns. For this purpose, we present an approach tailored for detecting RNA shape similarities. We extend the Structure to String (ST R2) method that was initially proposed to locate shape similarities in proteins to identify predicted secondary structures of RNAs. The ST R2 for RNAs is a translation of a secondary structure to a string of characters, after which known sequence-based search algorithms with an efficient implementation are being used. We validate that the ST R2 succeeds to locate G box riboswitches in prokaryotes, as expected. Subsequently we show running examples when attempting to detect G-box riboswitch candidates in eukaryotes. PMID- 15752076 TI - In silico reconstruction of nutrient-sensing signal transduction pathways in Aspergillus nidulans. AB - We report here probable nutrient-sensing signal transduction pathways in Aspergillus nidulans, a model filamentous fungus, based on sequence homology studies with known Saccharomyces cerevisiae and Schizosaccharomyces pombe proteins. Specifically, we identified A. nidulans homologs for yeast proteins involved in (1) filamentation-invasion, (2) cAMP-PKA, (3) pheromone response, (4) cell integrity and (5) TOR signaling pathways. We have also studied autophagy, one of the most important cellular responses regulated by TOR signaling. The Basic Local Alignment Search Tool program "blastp" was used to assess the homology of proteins. We note that by using a highly conservative approach, 70% of the S. cerevisiae signal transduction proteins (107 proteins out of 153 proteins studied) have significant homologs in A. nidulans. Using a slightly less conservative approach, we are able to identify homologs for as high as 91% of the S. cerevisiae signal transduction proteins (139 proteins out of 153 proteins studied). The filamentation-invasion, cell integrity and TOR signaling pathways showed greatest similarity with S. cerevisiae, while the cAMP-PKA and pheromone response pathways showed greater similarity with S. pombe. Based on these results, probable pathways in A. nidulans were constructed using well-established S. cerevisiae and S. pombe models. PMID- 15752077 TI - Mucosal diseases series. PMID- 15752078 TI - Epithelial biology. AB - The oral mucous membrane has features similar to skin but also differs in several ways. This paper reviews the aspects of epithelial biology necessary for an understanding of the vesiculoerosive disorders. PMID- 15752079 TI - Tongue disease in advanced AIDS. AB - OBJECTIVE: This study describes the involvement and the histological alterations found in the tongues of 92 autopsied patients who died with AIDS. MATERIALS AND METHODS: Sex, age, CD4 cell count and clinical history were obtained from the files of 92 patients who died with AIDS. All the tongues were examined for macroscopical alterations and stained using H&E, Gomori-Grocott, Ziehl-Neelsen, PAS, Brow-Hopps and Mucicarmine. Histological autopsy findings were grouped based on a protocol that was designed following the World Health Organization recommendations. RESULTS: The mean age of the patients who died of AIDS and CD4 cell count were 36 years and 82 cells microL(-1), respectively. Histological alterations of the tongues were found in 75% of the cadavers. The most common lesions were hairy leukoplakia (HL) (42 cases), candidosis (31 cases) and non specific chronic glossitis (29 cases), followed by concomitant lesions (28 cases), non-specific chronic ulceration (17 cases), melanotic pigmentation (13 cases), herpes simplex (10 cases), lymphoepithelial cysts (two cases), cryptococcosis (two cases), mycobacteriosis (one case), histoplasmosis (one case), cytomegalovirus infection (one case) and non-Hodgkin Lymphoma (one case). HL with oral candidosis (n = 13) were the most common concomitant lesions. CONCLUSION: These findings indicate that the tongue is a favorite site to occurrence of reactive, infectious and concurrent lesions in the end-stage of AIDS patients. PMID- 15752081 TI - The effects of chewing areca/betel quid with and without cigarette smoking on oral submucous fibrosis and oral mucosal lesions. AB - OBJECTIVES: The purpose of this study was to investigate the risk of areca/betel quid chewing with or without cigarette smoking on oral submucous fibrosis (OSF) and other oral mucosal lesions. METHODS: A stratified case-control study was designed. There were in total 102 patients with oral mucosal lesions or OSF (confirmed pathologically) in the case group. OSF (n = 62) and oral mucosal lesions (n = 62) in 102 subjects were separately analyzed for men and women investigating their risks. RESULTS: For OSF, people with both smoking and chewing habits had a statistically significant odds ratio (OR) 8.68 (95% CI = 1.87, 40.23). For the group of people with chewing habit only and without any lifetime cigarette smoking habit, the OR was 4.51 (95% CI = 1.20, 16.94). For other oral mucosal lesions, people with mixed habits and chewing only had also significant risks (OR = 8.37 and 3.95, respectively). For both OSF and other oral lesions, the ORs of mixed habits and chewing only were both higher in women than in men. CONCLUSIONS: The areca/betel quid used in Taiwan does not contain any tobacco product. The only way of areca/betel quid could synergize with any tobacco product is through cigarette smoking. A statistically significant association with oral mucosal lesions and OSF was still found in the group of areca/betel quid chewing only. PMID- 15752080 TI - Prevalence and distribution of oral lesions: a cross-sectional study in Turkey. AB - OBJECTIVES: The purpose of this study was to evaluate the prevalence and distribution of oral lesions (OLs) in Turkish population. SUBJECTS AND METHODS: In this cross-sectional study, 765 residents (F/M: 375/390) were selected by the cluster sampling method and examined according to WHO criteria. RESULTS: Excessive melanin pigmentation (6.9%) was the most common lesion in the study population. The tongue lesions observed in this study were as follows: fissured tongue (5.2%), varices (4.1%), hairy tongue (3.8%), geographic tongue (1.0%), atrophic tongue papillae (0.7%) and ankyloglossia (0.3%). The denture-related lesions were denture stomatitis (4.3%), suction irritation (0.8%), denture hyperplasia and torus palatinus as bony lesion (0.5%) and traumatic ulcers (0.3%). In regression analysis, being 65 years old and over was found as a statistically significant risk factor for the occurrence of pigmentation, fissured tongue, varices, hairy tongue, denture stomatitis and petechiae (P < 0.05). CONCLUSIONS: Pigmentation, fissured tongue and denture stomatitis were observed to be the most common lesions in Turkish population. Elderly population was a significant risk factor for occurrence of some OLs. PMID- 15752082 TI - Statherin levels in saliva of patients with precancerous and cancerous lesions of the oral cavity: a preliminary report. AB - OBJECTIVE: The aim of this study was to measure concentration of human salivary statherin in patients with oral cavity pathologies and salivary gland diseases. SUBJECTS AND METHODS: Levels of statherin were analysed with High Performance Liquid Chromatography (HPLC) in following groups of subjects: group A: 24 patients with neoplastic diseases of salivary glands, group B: 13 patients with inflammatory lesions of salivary glands, group C: 13 patients with precancerous and cancerous lesions of the oral cavity excluding salivary gland tumors, group D: 20 healthy volunteers (control group). RESULTS: Our preliminary data indicated a sensible reduction of the statherin level in the saliva of patients with precancerous and cancerous lesions of the oral cavity (group C) compared with the healthy subjects (group D). The statherin levels are not significantly reduced either in the inflammatory (group B) or in the salivary glands tumours (group A), compared with the healthy subjects (group D). CONCLUSION: Statherin could play a protective effect in oral cavity in association with its other functions. PMID- 15752083 TI - The effects of acute and chronic lithium treatment on rat submandibular salivation. AB - OBJECTIVE: Acute and chronic actions of lithium on salivation induced by agonists associated with receptor-linked hydrolysis of membrane inositol phospholipids (carbachol and phenylephrine) and by agonist linked to activation of adenylate cyclase (isoproterenol) were investigated. MATERIAL AND METHODS: In anaesthetized rats, submandibular salivation induced by intravenous injection of carbachol, phenylephrine and isoproterenol, was measured and expressed as volume of fluid (microl) elicited per 100 mg wet weight of each gland per minute. The experiments were repeated after acute and chronic treatment of lithium (7 mg kg(-1)). The results were analysed with unpaired t-test. RESULTS: Chronic, but not acute lithium treatment significantly decreases carbachol- and phenylephrine-induced salivation while isoproterenol-induced salivation was not changed neither after acute nor after chronic administration of lithium. CONCLUSION: The results suggest that hyposalivation during chronic lithium therapy could be mediated by alterations in the phosphatidylinositol cycle and a consequent lack of inositol after agonist stimulation. PMID- 15752084 TI - Calcium pyrophosphate dihydrate crystal deposition disease of the temporomandibular joint. AB - A case of a 74-year-old woman with calcium pyrophosphate dihydrate crystal deposition disease of the temporomandibular joint (TMJ) is presented. This disease rarely involves the TMJ and is not usually considered in the differential diagnosis of TMJ disorders. To our knowledge, only 23 cases have been reported in the literature and only four without any destructive changes of the condyle as in the present case. PMID- 15752085 TI - Bilateral intraosseous adenoid cystic carcinoma of the mandible: report of a case with lung metastases at first clinical presentation. AB - OBJECTIVE: Malignant epithelial tumours arising in the jaws are very rare. Adenoid cystic carcinoma (ACC) represents approximately 7.5% of all carcinomas and only a few cases of intraosseous (central) ACC have been reported in the literature. MATERIALS: The salient clinico-pathological features of a case of ACC, bilaterally occurring in the mandible of a young caucasian woman who also had lung metastases are reported to appropriately characterize such unusual lesions and discriminate them from other tumours that more commonly affect the mandible. RESULTS: The patient presented with a painful swelling of the right retro-molar area and paraesthesia of the ipsilateral lower lip and radiological investigations disclosed bilateral radiolucent lesions of the mandible with unequivocal signs of malignancy but without intra-lesional calcifications or association with teeth roots or cystic component. Conventional histological examination disclosed typical ACC with solid and cribriform growth patterns and extensive infiltration of the adjacent tissues. CONCLUSIONS: The diagnosis of intraosseous malignant salivary gland type neoplasms is very difficult in view of their rarity and lack of specific signs and mainly achieved after histological examination and complete clinico-radiological work up. As surgical treatment of the patient was not indicated, due to extensive neoplastic disease, the patient is being controlled with multimodal treatment, including chemo- and radiotherapy and is alive with persistent disease 3 years after the original diagnosis. PMID- 15752087 TI - Risk for developing cancer in Sweden after use of snuff. PMID- 15752086 TI - A case of chronic calcium pyrophosphate dihydrate crystal disease (tophaceous pseudogout) in the temporomandibular joint. AB - Pseudogout is a rare joint disease which is characterized by the presence of calcium pyrophosphate dihydrate crystals in the intraarticular and periarticular tissue. The crystals tend to attach to fibrocartilage tissue. Pseudogout principally affects the knee and wrist joints. Involvement of the temporomandibular joint (TMJ) is very rare. There have been <20 cases reported world-wide. Both acute and chronic manifestations have been described. We present here an unusual case that necessitated a high condylectomy. PMID- 15752090 TI - Proceedings of the Sixth International Conference on Breath Odor. PMID- 15752091 TI - Trimethylaminuria (fish-odour syndrome) and oral malodour. AB - A small but important percentage of oral malodour cases have an extra-oral aetiology and certain of these fall into the category of 'blood-borne halitosis'. Odoriferous substances generated within the body and transported to the lungs via the circulatory system may, if sufficiently volatile, leave with the exhaled air and impart a foetid odour to the breath. The aliphatic tertiary amine, trimethylamine, is such a volatile compound that is generated to excess in patients with a metabolic disorder known as trimethylaminuria (fish-odour syndrome). This article highlights this condition and draws attention to its potential role in the causation of recalcitrant oral malodour. PMID- 15752092 TI - In vitro models for oral malodor. AB - A model is a representation of some real phenomena and contains aspects or elements of the real system to be modeled. The model reflects (or duplicates) the type of behavior (or mechanisms) seen in the real system. The main characteristic of any model is the mapping of elements or parameters found in the system being studied (e.g. tongue dorsum biofilm in situ) on to the model being devised (e.g. laboratory perfusion biofilm). Such parameters include correct physico-chemical (abiotic) conditions as well as biotic conditions that occur in both model and reality. The main purpose of a model is to provide information that better explains the processes observed or thought to occur in the real system. Such models can be abstract (mental, conceptual, theoretical, mathematical or computational) or 'physical', e.g. in the form of a real disaggregated in vitro system or laboratory model. A wide range of different model systems have been used in oral biofilm research. These will be briefly reviewed with special emphasis on those models that have contributed most to knowledge in breath odor research. The different model systems used in breath odor research are compared. Finally, the requirements for developing an overall 'bad breath model' from considering the processes as a whole (real oral cavity, substrates in saliva, biotransformation by tongue microflora, odor gases in the breath) and extending this to the detection of malodor by the human nose will be outlined and discussed. PMID- 15752093 TI - Malodour in denture wearers: an ill-defined problem. AB - Denture plaque has not been studied to the same extent as dental plaque, and although there are many similarities in microbial composition, there are some significant differences. Denture-induced stomatitis is associated with poor denture hygiene, a more acidogenic plaque and the presence of Candida albicans. Obligate Gram-negative anaerobic micro-organisms, although present in denture plaque, have rarely been specifically investigated. Opportunist pathogens including coliforms and staphylococci have been isolated from dentures. Teeth adjacent to partial dentures are more susceptible to caries and periodontal diseases, perhaps due to an increased plaque buildup at the prosthesis/tooth interface. Little work has been published on malodour associated with dentures. The inert material provides a substratum for the plaque biofilm, which encompasses a range of odour-producing species. The microbiology of the tongue in denture wearers has not been specifically studied. Thus the nature, origin and extent of malodour in denture wearers is ill-defined, but many species capable of producing malodorous compounds are present. The wide age and health range presented by denture wearers further confounds investigation. There is a need for further work in the area, both for cosmetic- and health-associated reasons in the increasing elderly population. PMID- 15752094 TI - The rationale and potential for the reduction of oral malodour using Streptococcus salivarius probiotics. AB - The primary treatment for oral malodour is the reduction of bacterial populations, especially those present on the tongue, by use of a variety of antimicrobial agents or mechanical devices. However, shortly after treatment the problematic bacteria quickly repopulate the tongue and the malodour returns. In our studies, we have used a broadly-active antimicrobial (chlorhexidine) to effect temporary depletion of the oral microbiota and then have attempted to repopulate the tongue surface with Streptococcus salivarius K12, a benign commensal probiotic. The objective of this is to prevent re-establishment of non desirable bacterial populations and thus help limit the re-occurrence of oral malodour over a prolonged period. In this paper, we discuss why contemporary probiotics are inadequate for treatment of oral malodour and examine the rationale for selection of particular bacterial species for future use in the treatment of this condition. In our preliminary trials of the use of a chlorhexidine rinse followed by strain K12 lozenges, the majority (8/13) of subjects with confirmed halitosis maintained reduced breath levels of volatile sulphur compounds for at least 2 weeks. We conclude that probiotic bacterial strains originally sourced from the indigenous oral microbiotas of healthy humans may have potential application as adjuncts for the prevention and treatment of halitosis. PMID- 15752095 TI - Pilot study of a breath test for volatile organic compounds associated with oral malodor: evidence for the role of oxidative stress. AB - BACKGROUND: We performed a pilot study of a new method to identify the volatile organic compounds (VOCs) in breath associated with oral malodor, using gas chromatography and mass spectroscopy (GC/MS). METHODS: Oral cavity breath was collected from seven patients with oral malodor. Breath samples (150 ml) were concentrated onto sorbent traps and analyzed by GC/MS. RESULTS: Organoleptic scores ranged from 3.0 to 4.0 (mean = 3.3) on a scale of 0-5. Twenty-four of 30 (80.0%) of the most abundant oral malodor volatile organic compounds (OMVOCs) were alkanes and methylated alkanes. These VOCs are products of oxidative stress, generated by lipid peroxidation of polyunsaturated fatty acids in cell membranes. CONCLUSIONS: Increased oxidative stress in the oral cavity of patients with oral malodor may account for the increased risk of atherosclerosis, coronary heart disease and stroke associated with periodontal disease. The breath test for OMVOCs could potentially provide an objective new test for the assessment of oral malodor. PMID- 15752096 TI - Comparison of ninhydrin method of detecting amine compounds with other methods of halitosis detection. AB - OBJECTIVES: Halitosis is a result of overactivity of Gram-negative bacteria for which protein amino acids are the major source of energy. Therefore, statistical correlation between concentrations of volatile sulphur compounds (VSCs) and low molecular-weight amines should be expected. MATERIALS AND METHODS: Eighty-four patients suffering from halitosis and 40 healthy volunteers aged 20-62 years (average 39.7) participated in the study. In all subjects low-molecular-weight amines were evaluated by the ninhydrin method. Patients with halitosis were randomized into treatment groups. Zinc tablets, tablets and mouthwash containing chlorhexidine or lyophilized lactic acid-forming bacteria were used. RESULTS: Analysis showed that the level of amines was highest in subjects with halitosis (0.39, s.d. +/- 0.06, P < 0.001) and correlated significantly with results of VSC measurement and organoleptic scores (P < 0.001). Reduction of amine levels after treatment was statistically significant (P < 0.001). Three months following treatment the amine levels began to increase (0.37, s.d. +/- 0.05). The amine levels in healthy controls were lower (0.29, s.d. +/- 0.07) and remained at a stable level. CONCLUSION: The salivary amine levels significantly correlated with VSC levels and organoleptic scores. Therefore, the ninhydrin method of detecting salivary amines may be an alternative or additional method of diagnosing halitosis. This method may also be used to evaluate treatment efficacy. PMID- 15752097 TI - Effects of training on odor judges scoring intensity. AB - OBJECTIVE: This pilot study was intended to test whether a training protocol improved validity of odor judges (OJs), with or without experience, and whether odorant types differed in error proneness. METHODS: The OJs (four experienced, two inexperienced) completed a 4-phase training protocol based on the American Society of Testing and Materials standards (ASTM): (i) introduction to sensory scales, n-butanol reference, sniffing techniques; (ii) pretraining measurements; 20 samples of varying intensities of four unpleasant and three pleasant odorants; (iii) exercises assessing quality, intensity, ranking, and matching; and (iv) post-training measurements. MAIN OUTCOME MEASURES: Subjects' intensity scores were analyzed as the absolute difference from the 'true' intensity (ASTM n butanol standard) using repeated measures ANOVA. RESULTS: Training significantly (P = 0.02) reduced OJ errors. Experienced and novice judges did not differ in average errors (P = 0.99), or in improvement in error from pre- to post-training (P = 0.94). Improvement was consistent from pre- to post-training for all odorants except dimethylsulfide for which errors worsened (P = 0.01). Unpleasant and pleasant odorants differed (P = 0.006) in error. After removing water the effects of water control scores from the pleasant odorants, the difference was not significant (P = 0.26). CONCLUSIONS: The OJs improved in their ability to assess odor intensity irrespective of previous experience. Training is recommended for all OJs prior to research trials. PMID- 15752099 TI - Breath odor evaluation by detection of volatile sulfur compounds - correlation with organoleptic odor ratings. PMID- 15752098 TI - Use of n-butanol as an odorant to standardize the organoleptic scale of breath odour judges. AB - OBJECTIVE: The alcohol n-butanol has been recommended for use as a standard odorant by various groups for the training or standardization of breath odour judges and sensory evaluation panels. The objective of this study is to assess the use of n-butanol as a suitable odorant for organoleptic training of breath judges. METHODS: One judge with full smell acuity was trained in the method of organoleptic assessment using odorant solutions of main chemical classes (acids, amines, indole and sulphides) with the exception of alcohols. The subject was proficient in scoring odorant solutions, standard gas mixtures and human breath using the Rosenberg 0-5 organoleptic scale. A wide range of n-butanol solutions were prepared from 0 to 90 000 ppm and dispensed as replicate 12-ml volumes in Universal bottles (24 ml) leaving a headspace of 12 ml. Sets of odorants were prepared, labelled by code, randomized and presented to the judge in a completely blind fashion. The judge scored each concentration. This process was repeated on 32 occasions over a period of 12 weeks. Mean values of data for each determination for each concentration series were plotted against the log concentrations of odorant. Linear regression slope analysis was used to measure slope, the 95% CI of slope and the scatter of points (R2 value). Headspace concentrations of odorant were determined using gas chromatography (GC) analysis. RESULTS: The n-butanol regression slope gave a high R(2) value (0.971) and low scatter. However, the data did not correspond to other published work using an ASTM method where the range of recommended butanol concentrations was insufficient at both the high and low ends to determine the top and threshold. Moreover, headspace analysis using GC confirmed the published gas concentrations to be in error by a factor of 100. It was also observed that high concentrations of odorants were irritant causing desensitization if used for prolonged periods. CONCLUSION: The published method had erroneous headspace calculations listed and n-butanol could not be recommended as a training odorant because of its irritancy. PMID- 15752100 TI - Clinical effectiveness of a triclosan/copolymer/sodium fluoride dentifrice in controlling oral malodor: a 3-week clinical trial. PMID- 15752101 TI - Effect of a triclosan/PVM/MA copolymer/fluoride dentifrice on volatile sulfur compounds in vitro. AB - OBJECTIVE: The objective of the investigation was to document the in vitro efficacy of a triclosan/PVM/MA copolymer/fluoride (TCF) dentifrice against the formation of volatile sulfur compounds (VSC) as well as the growth of H2S producing bacteria. Clinical studies using organoleptic judges, gas chromatography, or a portable sulfide monitor have generally been employed in the assessment of treatments for the control of oral malodor. However, these studies are not appropriate for screening purposes because of the expense and time required. METHODS: An in vitro method was developed for the purpose of screening new compounds, agents or formulations for their ability to control VSC formation and for determining bio-equivalence of efficacy when implementing changes in existing formulations. The method combines basic microbiological methods, dynamic flow cell techniques and head space analysis. The in vitro VSC method was validated by comparing the efficacy of two dentifrices containing TCF with a control fluoride dentifrice as the TCF products have been clinically proven to control oral malodor. RESULTS: In the validation studies, the TCF-containing dentifrices were significantly better (P < 0.05) than the control dentifrice in inhibiting VSC formation and reducing H(2)S-producing bacteria. For example, when compared with baseline, the TCF dentifrices reduced VSC formation between 42 and 49% compared with the control dentifrice which reduced VSC formation 3%. There was no significant difference (P > 0.05) between the two TCF dentifrice formulations. CONCLUSION: Using an in vitro breath VSC model, it has been demonstrated that two variants of a dentifrice containing triclosan, PVM/MA copolymer and fluoride have efficacy that is significantly better than a control fluoridated dentifrice and that there is no significant difference between the triclosan/PVM/MA copolymer/fluoride dentifrice variants. PMID- 15752102 TI - Microbiological culture analysis of the tongue anaerobic microflora in subjects with and without halitosis. AB - OBJECTIVE: Determination of the microflora present on the tongue dorsum of subjects with and without halitosis using conventional microbiological culture methods. METHODS: Twenty-one halitosis and 20 control patients were recruited using a strict clinical protocol. Samples were collected from the posterior dorsum of the tongue using a sterile brush. Each sample was vortex mixed for 30 s and serial 10-fold dilutions to 10(-7) were carried out. Samples were plated onto fastidious anaerobe agar (FAA) and FAA enriched with vancomycin. These were incubated under anaerobic conditions for 10 days at 37 degrees C. Strict anaerobes were identified by metronidazole sensitivity and bacteria were identified to genus level by a combination of colony morphology, Gram staining and biochemical and enzymatic tests (rapid ID 32 A). RESULTS: The predominant species in test and control groups were Veillonella sp. and Prevotella sp. Greater species diversity was found in the halitosis samples compared with controls. The halitosis samples contained an increased incidence of unidentifiable Gram-negative rods, Gram-positive rods and Gram-negative coccobacilli. CONCLUSIONS: There was no obvious association between halitosis and any specific bacterial genus. The increased species diversity found in halitosis samples suggests that halitosis may be the result of complex interactions between several bacterial species. The role of uncultivable bacteria may also be important in contributing to this process. PMID- 15752103 TI - Clinical efficacy of a triclosan/copolymer/NaF dentifrice and a commercially available breath-freshening dentifrice on hydrogen sulfide-forming bacteria. AB - OBJECTIVE: The objective of this randomized, cross-over clinical study was to compare the 4-h and overnight effects of a triclosan/copolymer/fluoride (TCF-AF) dentifrice and a commercially available breath freshening (CBF) dentifrice containing fluoride for their ability to control H(2)S-forming bacteria. METHODS: Following a 1 week washout period of brushing with a regular fluoride dentifrice, subjects reported to the clinical site without performing oral hygiene, eating or drinking in preparation for baseline saliva sampling. After providing a baseline saliva sample, subjects were randomly assigned a test dentifrice and instructed to brush their teeth for 1 min, twice a day for 1 week. On the morning of the eighth day, subjects returned to the test site having refrained from oral hygiene, eating and drinking for overnight sampling. Subjects then brushed for 1 min with the assigned dentifrice, and returned for 2 and 4 h postbrushing evaluations. Following a second 1 week washout, subjects repeated the same regimen, but now using the other dentifrice. Oral micro-flora samples were collected by subjects rinsing with 10 ml of sterile water for 10 s. Each collected sample was serially diluted and plated in duplicate onto lead acetate agar. After incubating for 72 h, dark colonies were counted, expressed as log colony-forming units/ml, and reductions from baseline were calculated. RESULTS: Compared with baseline, the TCF-AF dentifrice reduced H2S-forming bacteria 0.82, 0.80 and 0.22 log units at the 2 and 4 h and overnight time points compared with 0.60, 0.43 and 0.07 log units for the CBF dentifrice. The TCFAF dentifrice was statistically significantly better (P < 0.05) than the CBF dentifrice at 4 h and overnight after brushing the teeth. CONCLUSION: The results of this randomized study indicate that the TCF-AF dentifrice is effective in controlling H2S producing bacteria. PMID- 15752104 TI - Quantitative detection of volatile sulfur compound- producing microorganisms in oral specimens using real-time PCR. AB - OBJECTIVE: It is well-known that some periodontopathic bacteria, especially Porphyromonas gingivalis, Fusobacterium nucleatum, Tannerella forsythia (formerly Bacteroides forsythus or Tan. forsythensis), and Treponema denticola, actively produce volatile sulfur compounds (VSCs), such as H2S and CH3SH. We previously reported a qualitative relationship between periodontopathic bacteria and VSCs; however, a quantitative analysis of periodontopathic bacteria in oral specimens is required for further characterization of the relationship between oral bacteria and VSCs. In this study, we report a real-time polymerase chain reaction (PCR) method for the quantitative analysis of VSC-producing bacteria in oral specimens. SUBJECTS AND METHODS: Specimens were collected from 22 patients who visited the Preventive Dentistry and Breath Odor Clinic of Kyushu Dental College. A real-time PCR assay using the TaqMan system, based on the 5'-3' exonuclease activity of Taq polymerase, was employed for the quantitative analysis of periodontopathic bacteria that produce VSCs. RESULTS: Using real-time PCR, we performed a quantitative analysis of P. gingivalis, F. nucleatum, Tan. forsythia, and T. denticola in the saliva, on the tongue coat, and in the subgingival plaque of patients with oral malodor. CONCLUSION: Real-time PCR using the TaqMan system can be used for the quantitative analysis of VSC-producing oral bacteria. PMID- 15752105 TI - Association between oral malodour and periodontal disease-related parameters in a population of 71 Israelis. AB - The aim of the present study was to determine whether oral malodour and periodontal disease parameters are associated with one another in 71 Israeli subjects (mean age 36.2 +/- 18.4; ages ranging from 15 to 65). Parameters measured included whole mouth odour judge scoring, Halimeter, OK to Kiss test, gingival index, plaque index and probing depth. Odour judge scores were significantly associated with Halimeter (r = 0.55; P < 0.001), as well as the OK to Kiss test (r = 0.52; P < 0.001). However, neither gingival index, plaque index nor probing depth was significantly associated with odour judge scores or Halimeter scores. Logistic regression analysis showed that both Halimeter and OK to Kiss scores factored significantly (P = 0.005 and 0.018, respectively, odds ratios 14.9 and 2.7, respectively) in predicting the severity of oral malodour. Results suggest that in the population studied, periodontal health and oral malodour are not associated with one another. PMID- 15752107 TI - The analysis of characteristics of elderly people with high VSC level. AB - OBJECTIVE: The purpose of this study was to examine the characteristics of elderly subjects who had objectionable levels of volatile sulfur compound (VSC). SUBJECTS AND METHOD: In 2002, a total of 115 85-year-old persons in Japan were subjected to oral examinations, tongue coat collections, measurements of VSCs levels inside the mouth using a portable gas chromatography (Oral Chroma, Abilit, Japan), and assessments of quality of life (QOL) using an SF-36 questionnaire. RESULTS: Sixty-six of the subjects were edentulous and 49 were dentulous. They were divided into two groups by VSC levels, those with oral malodor (both H2S > 112 ppb and CH3SH > 26 ppb; subjects with oral malodor, OM group; n = 7) and those without (n = 108). Our results showed that tongue coat deposit amounts and proportion of dentulous subjects were significantly higher in the OM group. Further, in an analysis of QOL, the SF-36 scores for vitality, social functioning and mental health were significantly higher in OM. CONCLUSION: We found that elderly subjects with oral malodor tended to be dentulous and had large deposits of tongue coating. However, oral malodor in the OM group subjects did not appear to cause a disadvantage in their social lives. PMID- 15752106 TI - Creation of oral care flavours to deliver breath-freshening benefits. AB - OBJECTIVE: Oral care products deliver breath freshening primarily via mechano chemical cleaning or by antimicrobial active systems. Dental flavours provide taste benefits, and freshen breath mainly by sensorial masking. We aimed to determine whether flavours could deliver breath freshening in products by inhibiting bacterial volatile sulphide compound (VSC) production. SUBJECTS AND METHODS: Flavour materials were screened for inhibition of hydrogen sulphide formation by Klebsiella pneumoniae in vitro, grouped by efficacy, and data provided to flavourists. Flavours were formulated to maximize the content of VSC effective ingredients and re-screened to confirm performance. Extensive, iterative testing of flavours identified reliable creative rules to deliver efficient inhibition of H2S generation. Breath-freshening flavours in whole products were then tested in-house in a 'breath freshness panel'. MAIN OUTCOME MEASURES: Malodour of panellists (not preselected for malodour score) was scored before and after product use, on the 'Rosenberg' 0-5 scale, together with residual flavour score, by extensively trained judges. Products were tested in double-blind, crossover studies, and results analysed using ANOVA. RESULTS AND CONCLUSIONS: Products flavoured using these rules delivered significantly greater breath freshening at 2 h than control products, and equivalent benefits to products containing 0.1% (w/w) triclosan or 0.2% (w/w) zinc sulphate. PMID- 15752108 TI - Halitosis and periodontal disease in subjects with mental disabilities. AB - OBJECTIVE: The aim of this study was to investigate the relationship between halitosis, presence of N-benzoyl-DL-arginine-2-naphthylamide (BANA) positive activity (indicative of Treponema denticola, Porphyromonas gingivalis, Tannerella forsythensis) and periodontal conditions in subjects with mental disabilities. MATERIALS AND METHODS: The population consisted of 17 Down syndrome (DS), 17 mentally retarded (MR) and 17 mentally healthy subjects (MH) - control group. A portable sulfide monitor was used to measure the concentration of volatile sulfur compounds (VSC) found in the mouth. Clinical parameters [Plaque Index (PlI), Probing Depth (PD), Bleeding on Probing (BOP)] were obtained from six reference teeth. Subgingival plaque samples for BANA Test were taken from the same six teeth and dorsum of the tongue. RESULTS: PlI and BOP were higher in DS than in MR and MH, respectively (P < 0.05). Mean PD was similar between DS and healthy groups but it was higher than the MR group. No significant differences were found among the three groups regarding the presence of BANA positive species, however the VSC levels were significantly lower in DS (97.23 ppb) than in MR (203.23 ppb) and MH individuals (180.53 ppb). CONCLUSIONS: Even though the presence of BANA positive activity was similar among DS, MR and MH subjects, halitosis seemed to be lower in the DS group. PMID- 15752109 TI - Subjective patients' opinion and evaluation of halitosis using halimeter and organoleptic scores. AB - OBJECTIVES: The aim of the study was to evaluate the level of interest in fetor ex ore among respondents, patients of the Department of Conservative Dentistry in Warsaw. MATERIALS AND METHODS: A questionnaire was completed by 295 patients, 202 females and 93 males aged 18-74 years (average 38.73 years). Each person was also examined for organoleptic score and volatile sulfur compounds (VSC) by Halimeter. Halitosis was diagnosed if the average level of VSC was > or =125 ppb and the organoleptic measurement using a 0-5 point scale was > or =2. Statistical analysis was performed using chi-square test. RESULTS: Incidence of halitosis was greatest in age ranges 25-34 years (29.68%) and 45-54 years (24.52%). Sixty-eight (43.87%) persons with diagnosed halitosis frequently reported having a problem with bad breath and only 5.81% persons with halitosis did not notice this problem (P < 0.001). Among 54 healthy persons three (5.56%) reported having a problem with halitosis (P < 0.001). CONCLUSION: A statistically significant correlation was found between clinical organoleptic diagnosis of halitosis and VSC level by halimeter. Subjective patients' opinion correlated well with objective evaluation of halitosis. PMID- 15752110 TI - Relationship between the SF-36 questionnaire and patient's satisfaction following halitosis therapy. AB - OBJECTIVE: To assess the health-related quality of life (HRQOL) of patients complaining of halitosis at their first visit and at a later time when their complaint had diminished following therapy, using a self-administered questionnaire, the Medical Outcome Study Short Form-36 (SF-36). The aim of this study was to examine the relationship between HRQOL of patients before and after self-reported disappearance of their complaint following oral hygiene improvements for halitosis. SUBJECTS AND METHODS: Seventy patients of our special clinic for halitosis served as subjects. At the first visit, each completed the SF-36 before determination of volatile sulfur compound (VSC) concentration in mouth air. After excluding dropouts, the same measurements were performed for subjects whose self-reported complaint had disappeared following oral hygiene therapy. RESULTS AND DISCUSSION: At the initial visit, SF-36 scale scores for general health, vitality, social functioning, role-emotion, and mental health were significantly lower when compared with the national averages in Japan. For subjects with self-reported disappearance of complaint, only social functioning rose significantly among SF-36 scores at the end of the study. These results suggest that an awareness of improvement in social life could be related to patient's satisfaction with halitosis oral hygiene therapy. PMID- 15752111 TI - Lethal photosensitization of oral pathogens via red-filtered halogen lamp. AB - OBJECTIVE OF INVESTIGATION: The ability of laser irradiation in the presence of photosensitizing agent to induce lethal effect on oral bacteria is well documented. We designed an in-vitro experiment to achieve phototoxic results on two common oral pathogens, using a high intensity, red filtered halogen lamp. Our goal was to determine the minimum duration of light exposure and drug dilution to achieve at least 50% reduction in bacteria counts. METHODS: Two common oral pathogens, Porphyromonas gingivalis and Prevotella intermedia were used in experiments. The source for light energy was a continuous working, high intensity, red filtered, halogen lamp (HL) with light transmitted through a flexible light guide over petri dishes containing live bacteria. Microorganisms were exposed to light for 5, 10 and 20 min. Methylene Blue (MB) in concentrations of 0.1, 0.075, 0.05, 0.025 and 0.01% was used as a photosensitizing agent. Light energy alone and MB alone was used as controls. RESULTS AND DISCUSSION: Optimum lethal photosensitization (50% or more bacteria killing) of oral pathogens was achieved using halogen light illumination for 5 min and longer with 0.05% MB or exposure to light for 20 min in the presence of 0.025 and 0.01% MB. Light exposure of 20 min in the absence of MB was not effective in killing bacteria. In the absence of light, MB at concentrations of 0.025 and 0.001% was not effective. Reduction of bacteria with the use of 0.05% MB alone was also insignificant. However, 0.075 and 0.1% MB, even in the absence of light was found to be bacteriocidal. CONCLUSIONS: Our in-vitro data indicate that we were able to achieve lethal photosensitization of two common oral pathogens with high intensity red filtered HL in the presence of diluted MB. In this era of increased incidence of antibiotic resistance, bacterial killing with laser or light energy in the presence of photosensitizing agent can prove to be a valuable treatment modality. PMID- 15752112 TI - Is transmission of bacteria that cause halitosis from pets to humans possible? AB - OBJECTIVE: Bacteria that cause halitosis naturally colonize the animal's mouth. The possibility of transmission of these bacteria from pets to human is an intriguing question. Therefore, the possible correlation between halitosis and regular contact of dental patients with pet animals was the aim of the presented study. MATERIALS AND METHODS: The study covered 84 patients suffering from halitosis and 40 healthy controls aged 20-62 years. Each person completed a questionnaire and was then examined for organoleptic score, volatile sulphur compounds (VSC) and evaluation of low molecular mass amines in saliva by a ninhydrin colorimetric reaction. Halitosis was diagnosed if the average level of VSC was 125 ppb and the organoleptic measurement using a 0-5 point scale was 2. Statistical analysis was performed using Wilcoxon's and chi-square tests. RESULTS: The results showed that among the halitosis group over 80% of patients had reported having pets in childhood and over 70% owned a pet at present. In the control group results are 47 and 40%, respectively. Analysis showed a significant correlation between frequency of halitosis and regular contact with pets at present (P < 0.001) or in childhood (P < 0.001). CONCLUSION: Pets (dogs, cats) owned in childhood or at present may transmit bacteria that cause halitosis. PMID- 15752115 TI - Identification of the immunodominant regions of the Em18 antigen and improved serodiagnostic specificity for alveolar echinococcosis. AB - The aim of this work was to identify the immunodominant regions of the Em18 antigen to improve the specificity in diagnosis of alveolar echinococcosis (AE). Two recombinant antigens ReEm18-1 and ReEm18-2, which have the same sequence except that nine amino acid residues are absent in ReEm18-2, were tested by ELISA and Western Blot (WB) for their diagnostic efficiency. Serological evaluation of the two antigens demonstrated that the sensitivity of both antigens was 95.5% in ELISA and WB, and the specificity was 93.6% and 95.7% in ELISA, and 81.4% and 82.9% in WB, respectively. Five more expression clones (EmS1-EmS5), which contain different regions of the Em18 sequence, were constructed for defining the immunodominant regions of the antigen. Fourteen monoclonal antibodies (mAbs) against ReEm18-2 antigen and the sera from different groups of patients were used to identify the epitope regions of the five antigen fragments. Results showed that the epitopes recognized by the mAbs are located in the N-terminal third of the sequence, but the immunodominant area recognized by native serum antibodies may be located further downstream (C-terminal) in the sequence. The nonspecific cross-reactivity is due to epitopes present in the C-terminal third of the sequence. The antigen fragments that contain the first two-thirds of the sequence have the same sensitivity to AE sera as those of the ReEm18-1 and ReEm18-2 antigens, but removal of the C-terminal third of the sequence improved the specificity of the assay from 93.6% to 99.3% (ELISA) and 81.4% to 90.7% (WB). We conclude that the necessary part of the ReEm18 antigen sequence for AE diagnosis is the N-terminal half to two-thirds of the entire sequence. PMID- 15752116 TI - Echinococcus multilocularis metacestode extract triggers human basophils to release interleukin-4. AB - Infections with parasitic helminths are associated with a T helper 2 (Th2) immune response and IgE production. The underlying mechanism, however, is only partially understood. Recently we have isolated a protein from extracts of Schistosoma mansoni eggs that triggers human basophils from non-sensitized donors to release interleukin-4 (IL-4), the key cytokine of a Th2 response. We called this protein IPSE (for IL-4-inducing principle from Schistosoma mansoni eggs). Supposing that IPSE-like IL-4-inducing activities might be a general principle shared among different helminth species, we investigated extracts from the cestode E. multilocularis for its effect on human basophils. Our results showed that extracts from metacestodes of E. multilocularis cause basophil degranulation, as well as the secretion of histamine, IL-4 and IL-13, in a dose-dependent manner. IgE stripping and resensitization of basophils indicated that the mechanism of IL 4 induction requires the presence of IgE on the cells. Since analogous properties have been demonstrated earlier for IPSE, we think that S. mansoni and E. multilocularis may induce a Th2 response in their hosts via a related mechanism, namely, by the induction of IL-4 release from basophils. PMID- 15752117 TI - Inflammatory antigens of Brugia malayi and their effect on rodent host Mastomys coucha. AB - The study was aimed at identifying pro- and anti-inflammatory cytokine releasing potential of Brugia malayi adult worm fractions and their role in filarial infection and pathogenesis. THP-1 cells were incubated with soluble somatic Brugia malayi adult worm extract (BmAS) and its Sephadex G-200 fractions BmAFI, BmAFII and BmAFIII and the effect of the fractions on parasitological, immunological and lymph node parameters was assessed in Mastomys coucha. BmAFII stimulated the pro-inflammatory TNF-alpha, IL-1beta and IL-6 release; IL-10 release was insignificant. Sensitization of animals with BmAFII and subsequent intraperitoneal implantation of worms enhanced CMI response. BmAFII also increased lymph node weight and cellularity, stimulated lymph node mast cells and eliminated intraperitoneally instilled worms. BmAFI stimulated several folds more release of IL-10, whereas TNF-alpha release was negligible. Sensitization with BmAFI elicited low CMI responses, moderately stimulated mast cells and facilitated survival of implanted adult parasites. Fifty percent of naive animals exposed to BmAFI showed oedematous lymph nodes and increased node weight. NCP bound molecules corresponding to BmAFI and II showed cytokine-stimulating potential in vitro. It is concluded that BmAFII is protective and stimulates pro inflammatory cytokines, whereas BmAFI facilitates parasite survival and stimulates IL-10. PMID- 15752118 TI - Recombinant SSP4 protein from Trypanosoma cruzi amastigotes regulates nitric oxide production by macrophages. AB - Acute infection with Trypanosoma cruzi is characterized by immunosuppression mediated by T cells and macrophages (Mphis). Nitric oxide (NO) production during the initial phase of acute infection might participate in the clearance of parasites by Mphis, whereas its overproduction during the late phase of acute infection would account for the immunosuppression observed. Trypanosoma cruzi molecules that might regulate the host responses have not been fully identified. Here, we demonstrate that active immunization with MBP::SSP4, a recombinant protein derived from a surface antigen specific of T. cruzi amastigotes (TcSSP4), was able to stimulate Ab production (IgG1, IgG2a, and IgG2b). On the other hand, MBP::SSP4 was able to stimulate NO production by peritoneal Mphis from BALB/c mice and Mphis from the J774 cell line. This effect was also observed at the level of inducible nitric oxide synthase (iNOS) detected by Western Blot. Furthermore, MBP::SSP4 was also shown to induce the expression of IL-1alpha, IL 6, IL-12, IFN-gamma, and TNF-alpha in normal animals, and IL-10 in immunized animals. In addition the protein MBP::SSP4 was able to bind to the surface of PMphis and J774 Mphis. These results suggest that TcSSP4 could modulate Mphi NO production and this may represent a mechanism participating in the immunoregulatory processes during Chagas' disease. PMID- 15752121 TI - Ultraviolet-free phototherapy. AB - BACKGROUND: Ultraviolet (UV) radiation phototherapy has been used for decades in the management of common skin diseases. On the other hand, UV radiation is a complete carcinogen and as a consequence, UV phototherapy is usually not used for the long-term management of children and young adults and in combination with topical or systemic immunosuppressants. PURPOSE: The therapeutic effectiveness of a new UV-free irradiation device in the treatment of patients with atopic hand and/or foot eczema was studied. METHODS: In a single-blinded trial 10 patients with atopic hand and/or foot eczema were treated with a sham irradiation device and with the new developed UV-free irradiation device DermoDyne during the following 4 weeks. All irradiation's lasted 30 min per treatment three times weekly. RESULTS: UV-free irradiation was found to induce a significant clinical improvement of atopic hand and foot eczema (P = 0.0001) in marked contrast to the sham-irradiation (P = 0.39). CONCLUSION: Our studies demonstrate that visible light can be successfully used for the treatment of patients with atopic eczema. PMID- 15752119 TI - Phosphatidylcholine-specific phospholipase C but not gamma interferon regulate gene expression and secretion of CC Chemokine Ligand-2 (CCL-2) by human astrocytes during infection by Toxoplasma gondii. AB - We have used human astrocytoma-derived cells to investigate the cellular responses of central nervous system cells to Toxoplasma gondii infection. At 24 h post inoculation, the secretion of CCL-2 (or Monocyte Chemotactic Protein-1) was augmented six-fold over the control. This secretion was down-regulated by D609, a specific inhibitor of phosphatidylcholine-dependent phospholipase C (PC-PLC), but not modulated by gamma interferon (IFN-gamma). Ribonuclease protection assay analyses showed significant down-regulation of CCL-2 mRNA production during infection by Toxoplasma gondii when cells were treated by D609. The mRNA levels of the seven other chemokines studied were not modified by D609. CCL-2 seems to contribute to the cell recruitment during human cerebral reactivation of Toxoplasma gondii. Cellular production of this CC chemokine during toxoplasmosis may be regulated by a PC-PLC-dependent pathway. PMID- 15752122 TI - PUVA-gel vs. PUVA-bath therapy for severe recalcitrant palmoplantar dermatoses. A randomized, single-blinded prospective study. AB - BACKGROUND/PURPOSE: In order to avoid unwanted effects of systemic psoralen and ultraviolet A (PUVA) therapy, various topical PUVA treatment modalities have been developed and are being increasingly used. However, up to now very few controlled studies comparing the therapeutic efficacy of different topical photochemotherapy modalities are available. Thus, the aim of our study was to compare the clinical efficacy of conventional PUVA-bath therapy to topical PUVA-gel therapy in patients with recalcitrant dermatoses of the palms and soles. METHODS: Twenty patients with severe palmoplantar dermatoses or localized psoriatic plaques were enrolled in our observer-blinded, randomized half-sided study. The treatment modalities compared were: (i) aqueous 8-methoxypsoralen (8-MOP)-containing gel plus broadband UVA irradiation (PUVA-gel therapy) and (ii) 8-MOP bath of the hands and/or feet plus broadband UVA (PUVA-bath therapy). RESULTS: On the body half, which was randomized to PUVA-gel therapy, the median Area and Severity Index for palmoplantar dermatoses (ASIppd) decreased from 28 (range 6-56) to 1.5 (range 1-37, P = 0.00) after a median 33 (13-49) irradiations compared with a reduction from 26.5 (range 6-52.5) to 1.5 (range 0-38, P = 0.00) for PUVA-bath therapy. Both improvements of ASIppd scores were found to be statistically significant, with no significant difference between PUVA-gel and PUVA-bath therapy. Severe phototoxic reactions such as strong erythema, blistering and/or pain were not observed in any patient. CONCLUSION: PUVA-gel therapy seems to be an effective therapeutic alternative to conventional PUVA-bath therapy in treating localized dermatoses of the palms and soles. The advantage of PUVA-gel therapy is reduced organizational efforts and expenses. PMID- 15752123 TI - UVA1 phototherapy for disseminated granuloma annulare. AB - BACKGROUND/PURPOSE: Disseminated granuloma annulare is a benign granulomatous skin disease of unknown etiology. Recently, UVA1 (340-400 nm) phototherapy has been found effective in a small series of four patients. The purpose of this two center study was to determine the rate and duration of clinical response to UVA1 phototherapy in a larger cohort of 20 patients with disseminated granuloma annulare. METHODS: Twenty patients with long-standing, stable disease (median 42 months, 95% CI 23-105) underwent UVA1 phototherapy. Sixteen patients were treated with a high-dose regimen (median single dose 110 J/cm2, 95% CI 103-121) and four patients with a medium-dose regimen (median single dose 50 J/cm2, CI 50-50). The clinical response was graded on a 5-point scale [0 = none, 1 = poor, 2 = moderate, 3 = substantial, 4 = (near) complete]. After cessation of therapy, patients with a clinical score of 3 or 4 were followed up to evaluate the duration of clinical improvement. RESULTS: At the end of treatment, five patients each had substantial improvement or (near) complete clearance. Another five patients had a moderate response, three patients were considered as poor responders and two patients as treatment failures. Out of the 10 patients with good or excellent response nine were available for follow up. Of these, two patients were still clear after 3 and 6 months, and seven patients relapsed after a median of 3 months (95% CI 1.68-6.46). CONCLUSIONS: UVA1 phototherapy provided good or excellent results in half of our 20 patients with disseminated granuloma annulare. In the majority of patients with a satisfactory response, however, discontinuation of treatment was followed by early recurrence of disease. PMID- 15752124 TI - Pharmacokinetics and clinical effects of mono-L-aspartyl chlorin e6 (NPe6) photodynamic therapy in adult patients with primary or secondary cancer of the skin and mucosal surfaces. AB - BACKGROUND/PURPOSE: Mono-L-aspartyl chlorin e6 (NPe6) is a photosensitizer that exhibits chemical purity, absorption at 664 nm wavelength and may be useful in photodynamic therapy (PDT). METHODS: This open label phase I clinical trial at the University of California, Davis Medical Center examined the pharmacokinetic properties of Npe6 and clinical response to PDT with this photosensitizer. A single intravenous dose of Npe6 was administered to 14 cancer patients with superficial malignancies (basal cell carcinoma = 22 lesions, squamous cell cancer = 13 lesions, papillary carcinoma = 14 lesions). Patients received one of five ascending doses (0.5 mg/kg (n = 4), 1.0 mg/kg (n = 3), 1.65 mg/kg (n = 3), 2.5 mg/kg (n = 3), or 3.5 mg/kg (n = 1)) 4-8 h prior to light activation. The total light dose (range 25-200 J/cm2) depended on the tumor shape and size. Light was delivered using an argon-pumped tunable dye laser. Serum NPe6 concentrations were measured over a 28-day period. The toxicity and cutaneous clinical efficacy of NPe6 were observed. RESULTS: Four weeks post-PDT, 20 of 22 basal cell carcinoma tumors (91%) showed a complete response. Eighteen of 27 other malignant cutaneous tumors showed a complete (n = 15/27, 56%) or partial (n = 3/27, 11%) response. Fewer non-responders were seen at an Npe6 dose level of 1.65 mg/kg or higher. Only 2 of 14 patients experienced an adverse event that was definitely related to NPe6 administration. Photosensitivity resolved within 1 week of NPe6 dosing in 12 of 14 patients. Analysis of serum levels of 11 individual patients indicated that a two-compartment model with a residual phase best fits the data. The mean alpha, beta, and terminal half-lives were 8.63+/-2.92, 105.90+/-37.59 and 168.11+/-53.40 h (+/-1 SD), respectively. The observed mean volume of distribution was 5.94+/ 2.55 l, and the mean clearance was 0.0394+/-0.0132 l/h. These values were independent of the dose administered. CONCLUSION: The photosensitizer, NPe6, was well tolerated with minimal phototoxic side effects, and demonstrated preliminary efficacy against cutaneous malignancies. PMID- 15752125 TI - No additional effect of topical calcipotriol on narrow-band UVB phototherapy in patients with generalized vitiligo. AB - BACKGROUND/PURPOSE: There is no definite cure for vitiligo; however, treatment responses with photobiological modalities are quite acceptable. Of all these, narrow-band UVB phototherapy was proposed rather recently. Calcipotriol has been shown to have stimulating activity on melanogenesis besides immunomodulatory and anti-inflammatory effects. This study was performed to determine whether adding topical calcipotriol to narrow-band UVB phototherapy enhances the efficacy of treatment. METHODS: In this prospective, single-blinded (investigator), right left comparison clinical study, 20 patients with generalized vitiligo were enrolled. Symmetrical lesions with similar sizes, bilaterally distributed on arms, legs, hands, feet or trunk were selected as reference lesions. In addition to narrow-band UVB, totally 96 treatment sessions, received two or three times weekly, the patients were asked to apply 0.005% topical calcipotriol on the selected side of the reference lesions twice daily. Then, they were monitored at the end of every 24-session interval. RESULTS: Cosmetically acceptable repigmentation was observed in 55% (11/20) of the patients without taking calcipotriol into account. There was statistically significant better response on the side that calcipotriol was not applied at the 24th session (P < 0.05). No statistically significant difference was found between the calcipotriol-treated and non-treated sides at 48th, 72th, and 96th sessions (P > 0.05). CONCLUSION: Our data confirm that, narrow-band UVB phototherapy is effective by itself in vitiligo, and show that adding topical calcipotriol does not improve treatment outcome. PMID- 15752126 TI - Ultraviolet protective performance of photoprotective lipsticks: change of spectral transmittance because of ultraviolet exposure. AB - BACKGROUND: Photoinstability of sunscreens because of ultraviolet (UV) exposure is a well-known and common phenomenon. Recently, it was also shown that sunscreens with complex filter combinations are photoinactivated by UV exposures, which can easily be acquired by solar exposure over several hours. OBJECTIVES: To assess the change of the spectral transmission after UV exposure (UV-challenged protective performance) of 27 commercially available photoprotective lipsticks. METHODS: Quartz slides were covered with a lipstick layer (area density 1.0+/-0.1 mg/cm2) and irradiated with increasing doses of solar-simulated radiation. The spectral transmission (T) was measured spectrophotometrically before and after 5, 12.5, 25, and 50 standard erythema doses (SED) of exposure. We calculated the change in transmission (photoinstability) as the difference between the spectral transmission before and after a defined UV exposure, DeltaT, and the arithmetic mean, for both the UVA (DeltaTA) and UVB (DeltaTB) ranges. A product was labelled as photounstable if the mean photoinstability in the UVA, DeltaTA, or UVB range, DeltaTB, was higher than 5% for an UV exposure of 12.5 SED. RESULTS: Eleven products showed a significant photoinstability in the UVA range (DeltaTA between 6% and 27%), only one product in the UVB range (DeltaTB = 13%), and one product in both the UVA (DeltaTA = 31%) and UVB (DeltaTB = 9%) range. In one product photoinstability became significant in the UVA range at higher UV exposures. CONCLUSIONS: Out of 27 lipsticks only 13 products showed a photostable performance (DeltaTA < 5% and DeltaTB < 5% for 12.5 SED). We propose therefore that only products, which fulfil these UV photostability criteria should be marketed. PMID- 15752127 TI - UVA-1 cold light therapy in the treatment of atopic dermatitis: 61 patients treated in the Leiden University Medical Center. AB - BACKGROUND: UVA-1 has been shown to be effective in the treatment of patients with atopic dermatitis. However, its optimal therapeutic conditions are not yet fully established. METHODS: In an open prospective study we retrospectively compared the effect of 4 weeks therapy (32 patients) with the effect of the usual 3 weeks therapy (29 patients) in patients with atopic dermatitis, using a medium dose UVA-1 cold light (45 J/cm2), 5 days a week. RESULTS: Scoring atopic dermatitis index (SCORAD) and dermatology life quality index (DLQI) quality of life indexes improved significantly during both 3 and 4 weeks UVA-1. Patients who were treated for 4 weeks showed a superior improvement of the SCORAD index [23.12 points, 95% confidence interval (CI) 16.09-30.16, vs. 13.32 points, 95% CI 5.61 21.04, P = 0.059], and the DLQI (5.41 points, 95% CI 2.38-7.88, vs. 3.86 points, 95% CI 1.88-5.84, P = 0.360), compared with patients who were treated for 3 weeks. However, the differences did not reach statistical significance. Only patients who were treated for 4 weeks were able to maintain their improvement 6 weeks after therapy. In both groups 50% of patients had intermittently used mild topical corticosteroids in the follow-up period. CONCLUSION: Extension of UVA-1 therapy from 3 to 4 weeks results in a clinically relevant improvement of the outcome, and more prolonged therapeutic effects, measured by the SCORAD index. PMID- 15752128 TI - Fixed solar urticaria induced by UVA and visible light: a report of a case. AB - Fixed solar urticaria (FSU) is a rare and less severe subgroup of solar urticaria. It is characterized by urticarial eruptions, which occurs on the same parts of the body following sun exposure. The lesions are reproducible at the same sites with similar morphology and distribution pattern after repeated sun exposure. The action spectrum of FSU is broad (300-700 nm). We reported a case of FSU induced by UVA and visible light. The patient responded well to cetirizine treatment. PMID- 15752129 TI - A case of toxic epidermal necrolysis with lesions mostly on sun-exposed skin. AB - Toxic epidermal necrolysis (TEN) is a cutaneous, mostly drug-induced reaction pattern characterized by a widespread necrosis and exfoliation of epidermis, involving more than 30% of the body surface area. Photo-induced Stevens-Johnson syndrome and erythema multiforme, which belong to the same spectrum of TEN, have been described before. However, there is only one case of TEN limited to sun exposed areas, which was caused by clobazam. In this report a second case of TEN because of naproxen, with lesions showing clear-cut borders on sun-exposed body areas resembling a sun-burn reaction, is presented. PMID- 15752130 TI - UVA-1 Phototherapy. PMID- 15752132 TI - Chronic actinic dermatitis recognized on minimal erythema dose testing prior to narrow-band UVB treatment for psoriasis. PMID- 15752133 TI - Crossing the divide--transport between the endoplasmic reticulum and Golgi apparatus in plants. AB - The transport of proteins between the endoplasmic reticulum (ER) and the Golgi apparatus in plants is an exciting and constantly expanding topic, which has attracted much attention in recent years. The study of protein transport within the secretory pathway is a relatively new field, dating back to the 1970s for mammalian cells and considerably later for plants. This may explain why COPI- and COPII-mediated transport between the ER and the Golgi in plants is only now becoming clear, while the existence of these pathways in other organisms is relatively well documented. We summarize current knowledge of these protein transport routes, as well as highlighting key differences between those of plant systems and those of mammals and yeast. These differences have necessitated the study of plant-specific aspects of protein transport in the early secretory pathway, and this review discusses recent developments in this area. Advances in live-cell-imaging technology have allowed the observation of protein movement in vivo, giving a new insight into many of the processes involved in vesicle formation and protein trafficking. The use of these new technologies has been combined with more traditional methods, such as protein biochemistry and electron microscopy, to increase our understanding of the transport routes in the cell. PMID- 15752134 TI - Exiting the endoplasmic reticulum. AB - Vesicular transport from the endoplasmic reticulum (ER) to the Golgi complex constitutes the initial step in protein secretion. COPII-coated vesicles mediate the export of newly synthesized proteins from the ER, and this transport step is coupled with COPI-mediated retrograde traffic to form a transport circuit that supports the compositional asymmetry of the ER-Golgi system. Biochemical and structural studies have advanced our understanding of the mechanisms that control vesicle formation and cargo-protein capture. Recent work has highlighted the function of transitional ER regions in specifying the location of COPII budding. PMID- 15752135 TI - The cellular pathway of CD1e in immature and maturing dendritic cells. AB - Dendritic cells (DCs) present antigens to T cells via CD1, HLA class I or class II molecules. During maturation, HLA class II-restricted presentation is optimized. The relocalization of CD1e from Golgi to endosomal compartments during DC maturation suggests also an optimization of the antigen-presentation pathway via CD1 molecules. We here detail the biosynthesis and cellular pathway of CD1e in immature and maturing DCs. Unlike the other CD1 molecules, CD1e was found to reach late endosomes through sorting endosomes, without passing through the plasma membrane in either immature or maturing cells. After induction of DC maturation, CD1e disappeared rapidly from the Golgi and was transiently localized in HLA-DR+ vesicles, while the number of CD1e+/CD1b+ compartments increased for at least 20 h. High-resolution light microscopy showed that, in immature DCs, CD1e+ vesicles were often in close apposition to EEA1+ or HLA-DR+ compartments, while CD1e displayed a nearly exclusive distribution in the lysosomes of mature DCs, a finding corroborated by immunoelectron microscopy. During maturation, CD1e synthesis progressively declined, while the endosomal cleavage of CD1e still occurred. Thus, CD1e displays peculiar properties, suggesting an unexpected role among the family of CD1 antigen-presenting molecules. PMID- 15752137 TI - Characterization of endocytic vesicles using magnetic microbeads coated with signalling ligands. AB - Iron microbeads coated with the protein ligands insulin and EGF (Fe-INS and Fe EGF) were prepared. Examination of the traffic of these ligand-coated microbeads demonstrated their internalization via clathrin-coated vesicles. Using magnetic methods, we have purified vesicles derived from the endocytic pathway. Vesicles prepared by this method are essentially free of contamination with other endomembrane compartments. Examination of the vesicles derived from cells treated with Fe-INS beads demonstrated the presence of the components of the Ras/Erk cascade on their surface. We conclude that the coupling of the Erk-signalling cascade induced by insulin takes place on the surface of endocytic vesicles derived from the internalization of the insulin receptor. PMID- 15752136 TI - Core formation and the acquisition of fusion competence are linked during secretory granule maturation in Tetrahymena. AB - The formation of dense core secretory granules is a multistage process beginning in the trans Golgi network and continuing during a period of granule maturation. Direct interactions between proteins in the membrane and those in the forming dense core may be important for sorting during this process, as well as for organizing membrane proteins in mature granules. We have isolated two mutants in dense core granule formation in the ciliate Tetrahymena thermophila, an organism in which this pathway is genetically accessible. The mutants lie in two distinct genes but have similar phenotypes, marked by accumulation of a set of granule cargo markers in intracellular vesicles resembling immature secretory granules. Sorting to these vesicles appears specific, since they do not contain detectable levels of an extraneous secretory marker. The mutants were initially identified on the basis of aberrant proprotein processing, but also showed defects in the docking of the immature granules. These defects, in core assembly and docking, were similarly conditional with respect to growth conditions, and therefore are likely to be tightly linked. In starved cells, the processing defect was less severe, and the immature granules could dock but still did not undergo stimulated exocytosis. We identified a lumenal protein that localizes to the docking competent end of wildtype granules, but which is delocalized in the mutants. Our results suggest that dense cores have functionally distinct domains that may be important for organizing membrane proteins involved in docking and fusion. PMID- 15752138 TI - Molecular basis for dissimilar nuclear trafficking of the actin-bundling protein isoforms T- and L-plastin. AB - T- and L-plastin are highly similar actin-bundling proteins implicated in the regulation of cell morphology, lamellipodium protrusion, bacterial invasion and tumor progression. We show that T-plastin localizes predominantly to the cytoplasm, whereas L-plastin distributes between nucleus and cytoplasm in HeLa or Cos cells. T-plastin shows nuclear accumulation upon incubation of cells with the CRM1 antagonist leptomycin B (LMB). We identified a Rev-like nuclear export sequence (NES) in T-plastin that is able to export an otherwise nuclear protein in an LMB-dependent manner. Deletion of the NES promotes nuclear accumulation of T-plastin. Mutation of residues L17, F21 or L26 in the T-plastin NES inhibits nuclear efflux. L-plastin harbors a less conserved NES and lacks the F21 T plastin residue. Insertion of a Phe residue in the L-plastin NES specifically enhances its export activity. These findings explain why both isoforms exhibit specific distribution patterns in eukaryotic cells. PMID- 15752139 TI - New faces of the familiar clathrin lattice. AB - The clathrin triskelion self-assembles into a lattice that coats transport vesicles participating in several key membrane traffic pathways. A new model of a clathrin lattice at approximately 8 angstrom resolution, generated by Fotin et al. (Nature 2004;432:573) confirmed the basic structural features of clathrin that were defined over many years of biochemical and structural analysis. In addition, new structural features of the clathrin trimerization domain were modelled for the first time, and the predictions correlated well with previous biochemical studies. A second model, placing auxilin within the lattice suggested a possible lattice contact targeted during lattice disassembly (Fotin et al. Nature 2004;432:649). This contact predicts interactions of the newly modelled trimerization domain with a newly defined extension of the clathrin triskelion, the ankle domain. These aspects of the new models were emphasized in the published reports describing them and in recent commentary (Brodsky, Nature 2004;432:568). Also emerging from the new models is a better picture of how the clathrin structure is distributed throughout the lattice, allowing the first predictions of interacting molecular interfaces contributing to contacts in the assembled lattice. The focus of this interchange is to emphasize these additional features revealed by the recently published models from Fotin and colleagues. PMID- 15752140 TI - Meeting report: EMBO workshop 'Cell Biology of Virus Infection', September 25-29, 2004, EMBL, Heidelberg, Germany. PMID- 15752142 TI - Transfusion medicine illustrated. A weapon of mass destruction: the spleen in immune thrombocytopenia. PMID- 15752143 TI - The dilemma of screening for antibodies against low-frequency human platelet antigens. PMID- 15752144 TI - Blood donor screening for agents of bioterror: more questions than answers. PMID- 15752145 TI - High variability of the RH locus in different ethnic backgrounds. PMID- 15752146 TI - Augmented mobilization and collection of CD34+ hematopoietic cells from normal human volunteers stimulated with granulocyte-colony-stimulating factor by single dose administration of AMD3100, a CXCR4 antagonist. AB - BACKGROUND: AMD3100, a selective antagonist of CXCR4, rapidly mobilizes CD34+ hematopoietic progenitor cells (HPCs) from marrow to peripheral blood with minimal side effects. STUDY DESIGN AND METHODS: To further investigate potential clinical utility of AMD3100 for CD34+ cell mobilization and collection, a Phase I study in normal volunteers was performed examining single-dose administration of AMD3100 alone and in combination with a standard 5-day granulocyte-colony stimulating factor (G-CSF) regimen. RESULTS: AMD3100 (160 microg/kg x 1 on Day 5) significantly increased both G-CSF-stimulated (10 microg/kg/day) mobilization of CD34+ cells (3.8-fold) and leukapheresis yield of CD34+ cells. Moreover, collection of CD34+ cells was comparable between individuals mobilized by a single-dose regimen of AMD3100 (240 microg/kg) and individuals mobilized with a 5 day regimen of G-CSF. AMD3100-mobilized leukapheresis products contained significantly greater numbers of T and B cells compared to G-CSF-stimulated leukapheresis products. CONCLUSION: These findings indicate that AMD3100 can be used alone or as an adjunct to G-CSF to mobilize cells for HPC transplantation. PMID- 15752147 TI - Minimum conditions of major histocompatibility complex compatibility and recipient immune compromise required to establish donor white blood cell persistence in a murine transfusion model. AB - BACKGROUND: In some patients multiply transfused to treat severe trauma, white blood cells (WBCs) from a single blood donor can persist for years, constituting up to 5 percent of all circulating WBCs. The immunologic mechanisms responsible for this are not known but, if understood, might allow manipulation of the human immune system to induce microchimerism for a variety of therapeutic purposes. To better characterize these mechanisms, a murine transfusion model was developed with a panel of immunologic knockouts as transfusion recipients. By conducting a systematic series of transfusion experiments, the purpose was to determine which recipient immune cell population, when abrogated, could lead to prolonged survival of donor cells (microchimerism). STUDY DESIGN AND METHODS: Blood was transfused from normal donors to knockout recipients in syngeneic, allogeneic, and xenogeneic settings. Donor WBC survival was evaluated by quantitative polymerase chain reaction, and recipient lymphocyte subsets by fluorescence activated cell sorting. RESULTS: In the syngeneic setting, donor WBCs persisted in C2ta, RAG-1, and TCR knockout recipients. Allogeneic donor WBCs persisted in RAG-2 and RAG-2/Common gamma knockout recipients. Xenogeneic donor WBCs required RAG-2/Common gamma and RAG-2/Pfp double knockouts to persist. CONCLUSION: It is concluded that donor-recipient major histocompatibility complex (MHC) concordance alone is not sufficient to achieve microchimerism. Further, the degree of recipient immune compromise necessary to achieve persistent microchimerism is directly proportional to the degree of donor-recipient MHC disparity. PMID- 15752148 TI - Phenotypic characterization and preclinical production of human lineage-negative cells for regenerative stem cell therapy. AB - BACKGROUND: Regenerative stem cell therapy (SCT) is currently being tested in clinical trials. The ideal type and source of cells have not yet been defined. Lineage (Lin) depletion is an experimental procedure capable of enriching all recently recognized SC types with regenerative potency. This study was performed to define a practicable monoclonal antibody (MoAb) cocktail for Lin depletion and to test whether clinical-scale Lin depletion is possible. STUDY DESIGN AND METHODS: MoAbs (CD2/14/15/19/41/56/glycophorin A) were selected to mark seven mature hematopoietic lineages. Lin7-negative (Lin7NEG) cells were analyzed in peripheral blood (PB, n = 9), mobilized PB (MPB, n = 5), umbilical cord blood (UCB, n = 5), and marrow aspirates (BM, n = 4) by flow cytometry. Preclinical Lin depletion was tested with leukapheresis products from PB following good manufacturing practice (GMP) principles. RESULTS: Lin7NEG cells comprised 0.23 +/ 0.04, 0.27 +/- 0.03, 0.53 +/- 0.07, and 0.49 +/- 0.03 percent of PB, MPB, UCB, and BM, respectively. Basophils, CD34+, and dendritic cells constituted the major Lin7NEG subpopulations (84 +/- 2, 90 +/- 3, 40 +/- 3, and 80 +/- 3% in PB, MPB, UCB, and BM, respectively). Minor populations included CD7- and CD45- cells. Preclinical CD2/14/15/19/56 (Lin5) depletion after automated red blood cell and platelet reduction resulted in up to a 16.7-fold enrichment of CD34+ and CD34 /Lin5NEG cells. CONCLUSIONS: A seven-MoAb cocktail is sufficient to label more than 99 percent of nucleated cells in PB, MPB, UCB, and BM. Preclinical Lin depletion can be performed under GMP conditions from PB apheresis procedures. PMID- 15752149 TI - The highly variable RH locus in nonwhite persons hampers RHD zygosity determination but yields more insight into RH-related evolutionary events. AB - BACKGROUND: Knowledge about paternal RHD hemi- or homozygosity is of clinical interest in alloimmunized pregnant women. D negativity in white persons is usually caused by deletion of the RHD gene. Recently, the physical structure of the RH locus and the mechanism causing the deletion of the RHD gene have been explored, enabling RHD zygosity determination in white persons by specific detection of a hybrid Rhesus box characteristic for the RHD- locus. STUDY DESIGN AND METHODS: RHD zygosity was determined in 402 samples from five different ethnic groups by polymerase chain reaction (PCR)-restriction fragment length polymorphism and by a newly developed real-time quantitative PCR. The Rhesus boxes of samples showing discrepancies between both tests were cycle sequenced. RESULTS: In nonwhite persons, several mutated Rhesus boxes exist that hamper zygosity determination by detection of the RHD- locus. Such mutated Rhesus boxes in D+RHD homozygous black persons have a frequency of 0.22. In white persons, no mutated Rhesus boxes were encountered so far. CONCLUSIONS: Owing to the high frequency of the mutated Rhesus boxes, zygosity determination by detection of the RHD- locus is not feasible in nonwhite persons. The cosegregation of variant RHD genes (RHDpsi and (C)cdes) with specific mutated Rhesus boxes yields more insight into the evolutionary events concerning variant RHD genes and mutated Rhesus boxes. PMID- 15752150 TI - Genetic mechanisms of Rhesus box variation. AB - BACKGROUND: The RHD gene is flanked by two highly homologous DNA segments of approximately 9000 bp, the upstream and downstream Rhesus boxes. In haplotypes with an RHD deletion, the fusion of the two Rhesus boxes generates the single hybrid Rhesus box, the detection of which has been applied for RHD zygosity determination. Aberrant Rhesus boxes can confound this application and appear to be frequent among African individuals. STUDY DESIGN AND METHODS: A total of 5850 bp of the upstream and of the downstream Rhesus boxes were sequenced in 18 samples that were representative for all four D clusters and of the hybrid Rhesus boxes in four samples that were mistyped in assays for the hybrid Rhesus box. RESULTS: The known differences between upstream and downstream Rhesus boxes were in part restricted to subsets of RHD alleles. Forty-six additional polymorphisms were detected and caused by single-nucleotide substitutions, short insertions, or deletions. Gene conversions were found in the upstream Rhesus boxes of RHDpsi, DAU-1, and DAU-3 and in the downstream Rhesus boxes of Ccdes, weak D type 4.1, type 4.2 (DAR), and DAU-0. Recombinations between haplotypes were likely in several alleles like DIII type 4. Four nonstandard hybrid Rhesus boxes were suggestive of multiple RHD deletion events. CONCLUSION: There is considerable variation of Rhesus box sequences associated with distinct RHD alleles. RHD zygosity diagnostics in African persons is best based on quantitative polymerase chain reaction or amplification of the full-length hybrid Rhesus box. Because aberrant Rhesus boxes were observed among European persons, use of more than one method for hybrid Rhesus box detection may even be advisable in European persons. PMID- 15752151 TI - Molecular characterization of D- Korean persons: development of a diagnostic strategy. AB - BACKGROUND: D- frequencies show wide racial differences: in particular, in Korean persons it is approximately 1/100th of that in Caucasians (0.15% vs. 15%). The molecular mechanisms of D- may be unique to races, and thus specific molecular diagnostic approaches are necessary for individual races. Such marked racial differences have been attributed to the founder effect. It was hypothesized that D- frequencies may be affected by genomic variations of Rhesus boxes, which are instruments of unequal crossing-over. STUDY DESIGN AND METHODS: First, the molecular basis of D- in Korean donors was characterized. A total of 264 D- persons were analyzed by performing RHD exon polymerase chain reaction (PCR), Rhesus box hybrid PCR-PstI, RHD sequencing, and a novel RHD-CE-D hybrid PCR. Second, with sequencing analysis, Rhesus boxes in Korean and Caucasian persons were compared. RESULTS: Of 264 D- Korean individuals, 74 percent completely lacked RHD, 9 percent had RHD-CE-D hybrid, and 17 percent had point mutations. Three genetic causes, RHD deletion, RHD-CE(2-9)-D2, and 1227G>A, explained 99.4 percent of D- alleles in Korean persons. Three novel mutations were also found. The identity region of 10 Rhesus boxes in Korean persons was in complete concordance with that in Caucasian persons. CONCLUSION: In this study, a molecular diagnostic strategy was established and the genetic causes of almost all D- Korean persons was able to be diagnosed with a simple decision tree. The study provides a good example of the molecular diagnosis of D- persons, especially in low-frequency areas. Our hypothesis that D- frequencies are affected by genomic variations in Rhesus boxes was excluded. Rather, racial differences may be influenced by the founder effect. PMID- 15752152 TI - Immunization against a low-frequency human platelet alloantigen in fetal alloimmune thrombocytopenia is not a single event: characterization by the combined use of reference DNA and novel allele-specific cell lines expressing recombinant antigens. AB - BACKGROUND: Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is caused by maternal immunization against a fetal platelet (PLT) alloantigen. In cases of FNAIT attributed to low-frequency PLT alloantigens, the laboratory diagnosis is often hampered by the lack of adequate PLTs. STUDY DESIGN AND METHODS: Three families with maternal immunization against fetal PLT antigens were analyzed. In Family 1, previous immunization of another female or woman has been observed. In Families 2 and 3, newborns presented with the typical clinical picture of FNAIT. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing with reference to DNA from Epstein-Barr virus transformed B-lymphoblastoid cell lines. Antibodies were characterized by glycoprotein (GP)-specific immunoassay with a panel of stable Chinese hamster ovary cell lines expressing low-frequency alloantigens. RESULTS: In three families, maternal immunization associated with the low-frequency alloantigens human PLT antigen (HPA)-8bw (Sra), HPA-11bw (Groa), and HPA-13bw (Sita) was identified. Maternal serum samples showed positive reactions in an antigen capture assay with cell lines carrying recombinant GP IIb/IIIa (HPA-8bw and 11bw) or GPIa/IIa (HPA-13bw), respectively. These results could be confirmed by genotyping analysis of fathers and newborns. CONCLUSION: This study demonstrates that cases of FNAIT attributed to low-frequency PLT alloantigens cannot be regarded as single events. The availability of reference DNA and cell lines expressing recombinant PLT alloantigens can facilitate their identification. PMID- 15752153 TI - Nondeletional ABO*O alleles express weak blood group A phenotypes. AB - BACKGROUND: Owing to a single-base deletion, the vast majority of ABO*O alleles encode for a truncated and catalytically inactive ABO glycosyltransferase, leading to the generation of a premature stop codon. Less frequent nondeletional ABO*O alleles such as ABO*O03, in contrast, have nonsynonymous mutations that may abolish the protein's enzyme activity by altering its sugar-binding site. STUDY DESIGN AND METHODS: Extensive ABO phenotyping and genotyping were performed in healthy blood group O donors with weak anti-A isoagglutinins and their relatives as well as in blood group O donors selected for the presence of ABO*O03. HeLa cells were used to transfect ABO expression plasmids. RESULTS: Donors or relatives carrying ABO*O03 and/or its rare variant ABO*Aw08 in homozygous (n = 2) or heterozygous (n = 14) form showed weak A antigen expression detectable only by adsorption-elution (n = 15) or by monoclonal anti-A typing (n = 1). The serum samples of most donors (n = 13) contained weak anti-A; in the remaining donors, anti-A isoagglutinin reactivity was in the normal range. In the transfection studies, weak A antigen expression on HeLa cells transfected with plasmids containing ABO*O03 or ABO*Aw08 expression constructs was detectable only by adsorption-elution. CONCLUSION: The data provide evidence that nondeletional ABO*O03-like alleles produce detectable amounts of A antigens. PMID- 15752154 TI - Relevance of the HPA-15 (Gov) polymorphism on CD109 in alloimmune thrombocytopenic syndromes. AB - BACKGROUND: Alloantibodies against the human platelet (PLT) alloantigen (HPA)-15 system residing on CD109 can cause fetal and neonatal alloimmune thrombocytopenia (FNAIT), posttransfusion purpura, and PLT transfusion refractoriness. The detection of antibodies against HPA-15, however, is hampered by the variable low expression and instability of the CD109 molecule during preparation and storage. STUDY DESIGN AND METHODS: This study analyzed the occurrence of HPA-15 alloantibodies in 1403 patients: 930 FNAIT and 473 polytransfused (PT) patients by modified monoclonal antibody specific immobilization of PLT antigens (MAIPA) assay with well-defined phenotyped PLTs. A DNA typing technique was developed to confirm the phenotypes of PLT donors. B-cell lines were established as sources of reference DNA. RESULTS: Genotyping of 407 unrelated blood donors revealed the gene frequencies 0.512 and 0.488 for HPA-15a and -15b, respectively. Based on the selection of PLTs expressing high amounts of CD109 on the surface (mean fluorescence intensity ratio 4-5 on expression peak on Days 2-4 after apheresis) antibody screening by the MAIPA assay was performed. In total, 16 (1.1%) HPA-15 alloantibodies were found comprising four anti-HPA-15a and 12 anti-HPA-15b. Anti HPA-15b without other PLT-reactive antibodies were detectable in three serum samples of PT patients. The incidence of HPA-15 alloimmunization in PT patients was significantly higher than in mothers with FNAIT (3.0% vs. 0.22%). In relation to all detected HPA-specific antibodies, HPA-15 is responsible for 6.2 percent of alloimmunizations. CONCLUSION: These observations indicate that alloimmunization against HPA-15 should be considered as a cause for immune thrombocytopenia, particularly in patients receiving multiple PLT transfusions. PMID- 15752155 TI - Surface decoration of red blood cells with maleimidophenyl-polyethylene glycol facilitated by thiolation with iminothiolane: an approach to mask A, B, and D antigens to generate universal red blood cells. AB - BACKGROUND: The surface decoration of red blood cells (RBCs) by polyethylene glycol (PEG) chains has been an approach developed to camouflage the blood group antigens from their antibodies. A PEGylation protocol, however, that can mask the antigens appropriately to inhibit the agglutination of RBCs with the respective antibodies is not available so far. STUDY DESIGN AND METHODS: A new approach for PEGylation of RBC membrane proteins has been designed with thiolation-mediated maleimide chemistry. The accessibility of the surface lysine residues of membrane proteins to bulky PEG reagents was increased by linking an extension arm carrying a thiol group. RESULTS: RBCs have been PEGylated by thiolation-mediated chemistry with maleimidophenyl-PEG (Mal-Phe-PEG) reagents of different chain lengths. Mal Phe-PEG-5000 chains alone masked the most important antigens of the Rh system (C, c, E, e, and D) from their antibodies. The masking of the A and B antigens needed a combination of Mal-Phe-PEG-5000 and Mal-Phe-PEG-20000 chains to inhibit the agglutination of RBCs completely with anti-A or anti-B. CONCLUSIONS: Thiolation mediated PEGylation of RBCs with Mal-Phe-PEG-5000 and Mal-Phe-PEG-20000 converts Group A Rh(D)+ and B Rh(D)+ RBCs into RBCs with serologic behavior comparable to Group O Rh(D)- RBCs that are considered as universal RBCs for transfusion. PMID- 15752156 TI - Chemical compounds that target thiol-disulfide groups on mononuclear phagocytes inhibit immune mediated phagocytosis of red blood cells. AB - BACKGROUND: Patients having immune cytopenias produce antibodies that target hematopoietic cells resulting in their phagocytosis and intracellular destruction. Early reports suggested that phagocytosis could be inhibited by interfering with membrane thiol (SH) groups on phagocytes. Thus, whether chemical compounds that interact with SH or disulfide (SS) groups on mononuclear phagocytes can inhibit phagocytosis of antibody-coated cells was examined. STUDY DESIGN AND METHODS: A monocyte monolayer assay (MMA), which examines the in vitro monocyte-macrophage (Mphi) interaction with anti-Rh(D)-coated red cells (RBCs), was used to study the ability of different SH and SS chemicals to inhibit the Fc receptor-mediated phagocytosis of sensitized RBCs. The compounds examined included thimerosal, dithiothreitol (DTT), pentane-1-thiol, and two recently described SH and two SS chemicals that have been synthesized. RESULTS: All compounds were found to be able to inhibit phagocytosis to varying degrees correlating to the structure of the molecule. In general, those compounds that interact with free SH groups to inhibit phagocytosis were found better than SH containing compounds that interact with SSs. Thimerosal and p-nitrophenyl methyl disulfide were the most effective compounds inhibiting phagocytosis. Both chemicals showed greater than 50 percent inhibition at concentrations as low as 10(-9) mol per L. DTT was the least effective compound tested. Only thimerosal showed significant toxicity, as determined by decreased cell viability and increased apoptosis, but only at concentrations of 10(-8) mol per L. The effect of chemical treatment was on attachment rather than on phagocytosis itself. Fcgamma receptor-independent endocytosis was not affected by the chemical treatment. CONCLUSION: These studies indicate that pharmacologic strategies that target SH groups on mononuclear phagocytes may have future efficacy for the treatment of immune cytopenias. PMID- 15752157 TI - Autologous peripheral blood progenitor cells are a potential source of parvovirus B19 infection. AB - BACKGROUND: Parvovirus B19 is a cause of delayed red blood cell (RBC) engraftment after marrow transplantation (BMT). The diagnosis of parvovirus infection requires serologic and DNA testing in the context of clinical disease and characteristic marrow morphologic findings; however, the source of infection is often difficult to determine. STUDY DESIGN AND METHODS: Investigation of a case of delayed RBC engraftment and pure RBC aplasia (PRCA) occurring 3 months after autologous peripheral blood progenitor cell (PBPC) transplantation in a patient with high-risk diffuse large B-cell lymphoma. DNA testing of serum and of a sample of cryopreserved PBPCs was performed. RESULTS: Marrow morphology showed a maturational arrest of erythroid cells with giant proerythroblasts. Polymerase chain reaction and nucleic acid hybridization confirmed the presence of parvovirus DNA in the serum and in a sample of sequestered PBPCs saved at the time of PBPC harvest. PRCA resolved after the administration of intravenous immune globulin. CONCLUSION: Autologous PBPCs are a potential source of parvovirus infection, which may cause significant disease after autologous BMT. PMID- 15752158 TI - Nucleic acid test screening of blood donors for orthopoxviruses can potentially prevent dispersion of viral agents in case of bioterrorism. AB - BACKGROUND: Microbiologic agents such as variola virus (VAR) are very attractive for terrorism. As a result of international collaboration under the WHO eradication campaign, smallpox was declared eradicated in 1980. Therefore, the immunization programs were discontinued worldwide. Because most people are now immunologically naive, VAR is considered to be a potential threat agent or bioterrorist weapon. Real-time polymerase chain reaction (PCR) followed by melting analysis was developed for fast and safe analysis and allows differentiation of VAR from other orthopoxviruses (OPVs) like vaccinia or camelpox virus. STUDY DESIGN AND METHODS: A RealArt Orthopox LC PCR kit (Artus GmbH) was used to amplify OPV sequences from blood donor samples. A total of 31,500 blood donor samples were tested in minipools of up to 96 samples. To evaluate the sensitivity of the assay, routine donor minipools (90 +/- 6 samples per pool) were spiked with vaccinia virus used as positive control. RESULTS: Specificity was 100 percent because none of 31,500 blood donors was positive for the presence OPV. The detection limit of the assay was 10.6 copies per PCR procedure. Therefore, a sensitivity of 1590 copies per mL was calculated. Overall, 0.28 percent of test results had to be considered invalid owing to negative internal controls. CONCLUSION: The RealArt Orthopox LC PCR kit enables reliable detection of OPV DNA in viremic blood donor samples, even at the beginning of the disease when patients present minor clinical symptoms, and could be implemented in our routine screening procedure immediately. Thus, the assay could potentially help to prevent dispersion of viral agents by blood transfusion in case of bioterrorism. PMID- 15752159 TI - The impact of male-to-male sexual experience on risk profiles of blood donors. AB - BACKGROUND: Men who have had sex with men (MSM) since 1977 are permanently deferred from donating blood. Excluding only men who engaged in male-to-male sex within either the prior 12 months or 5 years has been proposed. Little is known about infectious disease risks of MSM who donate blood. STUDY DESIGN AND METHODS: Weighted analyses of data from an anonymous mail survey of blood donors were conducted to examine the characteristics of men reporting male-to-male sex during specified time periods. RESULTS: Of the 25,168 male respondents, 569 (2.4%) reported male-to-male sex, 280 (1.2%) since 1977. Compared to donors who did not report male-to-male sex, the prevalence of reactive screening test results was higher among donors who reported the practice within the past 5 years (< or =12 months odds ratio [OR] 5.3, 95% confidence interval [CI] 2.6-10.4; >12 months to 5 years, OR 7.1, 95% CI 1.2-41.7); however, no significant difference was found for donors who last practiced male-to-male sex more than 5 years ago (>5 years after 1977, OR 1.4, 95% CI 0.7-2.6; 1977 or earlier, OR 1.6, 95% CI 0.7-3.7). The prevalence of unreported deferrable risks (UDRs) other than male-to-male sex was significantly higher for all donors who reported male-to-male sex with ORs ranging from 3.1 to 18.9 (p < or = 0.01). CONCLUSIONS: No evidence was found to support changing current policy to permit donations from men who practiced male to-male sex within the past 5 years. For donors with a more remote history of male-to-male sex, the findings were equivocal. A better understanding of the association between male-to-male sex and other UDRs appears needed. PMID- 15752160 TI - Plateletpheresis does not cause long-standing platelet-derived growth factor release into the donor blood. AB - BACKGROUND: Recently, long-standing elevations of soluble growth factors released from platelets (PLTs) after contact with artificial surfaces during dialysis were described. They could be jointly responsible for the high frequency of death from cardiovascular diseases in dialysis patients. There are no comparable data on the extent and the duration of a growth factor release by plateletpheresis procedures. STUDY DESIGN AND METHODS: A total of 37 plateletpheresis procedures were performed with two different devices. PLT-derived growth factor (PDGF) isoform AB, transforming growth factor (TGF)-beta1, and beta-thromboglobulin (beta-TG) were measured in the donors' plasma samples, and PLT activation and function were measured by cytometry and aggregometry before and after plateletpheresis and 1 and 24 hours later. RESULTS: Before apheresis, the following mean plasma levels were found: beta-TG, 98.6 +/- 37.3 IU per mL; PDGF AB, 71.5 +/- 38.5 pg per mL; and TGF-beta1, 2.24 +/- 0.80 ng per mL. At the end of the apheresis procedures, the mean PDGF-AB level had increased by a factor of 1.8 (p < 0.05). One hour later, the mean PDGF-AB level had normalized again. No significant change in the levels of beta-TG and TGF-beta1 was found by the apheresis procedures. There was no influence of the blood cell separator type on the results. CONCLUSION: Only a slight and rapidly reversible increase in soluble PDGF-AB was found during plateletpheresis and no increase in soluble TGF-beta1 and beta-TG was found. This change should not be harmful to the donor. PMID- 15752161 TI - Additive effects between platelet concentrates and desmopressin in antagonizing the platelet glycoprotein IIb/IIIa inhibitor eptifibatide. AB - BACKGROUND: Platelet (PLT) glycoprotein (GP) IIb/IIIa receptor antagonists have demonstrated efficacy in decreasing ischemic complications of percutaneous coronary intervention and/or unstable angina. In case of bleeding, the drug can be stopped and PLT transfusions can be given. STUDY DESIGN AND METHODS: This crossover study tested the additive effects of PLT concentrates (PCs) after desmopressin (DDAVP) infusion in antagonizing the anti-PLT effects of GPIIb/IIIa inhibitors and aspirin. After eptifibatide and aspirin infusion (at standard dosages), 10 healthy volunteers received DDAVP or placebo. Thereafter, increasing amounts of PLTs from fresh single-donor apheresis concentrates were added in vitro to blood samples of all volunteers to increase PLT counts by 30 x 10(9), 60 x 10(9), or 120 x 10(9) per L. RESULTS: Adding platelets in vitro further improved PLT function after DDAVP: it shortened collagen-adenosine diphosphate closure times (p < 0.01), to normal ranges as measured by the PLT function analyzer (PFA-100). In contrast, normal PLT function could not be restored even when PLT counts were increased by 50 percent (120 x 10(9)/L) in the placebo group. CONCLUSION: Combined use of PLTs from fresh apheresis PC and DDAVP additively enhances recovery of normal PLT function after eptifibatide infusion. Such a strategy may help to avoid excessive transfusion of PC. PMID- 15752162 TI - The quality of plasma collected by automated apheresis and of recovered plasma from leukodepleted whole blood. AB - BACKGROUND: There exists a current lack of information about the composition of the different types of plasma. No direct comparisons between apheresis plasma (AP) and recovered plasma (RP) derived from in-line-filtered whole blood (WB) have been published to date. STUDY DESIGN AND METHODS: Sixty AP units, 100 RP units from in-line-filtered WB held for 3 hours at 20 degrees C between donation and freezing, and an additional 100 RP units held for 15 hours at 20 degrees C before freezing were analyzed for coagulation factors and inhibitors, total protein, immunoglobulin G (IgG), and hemostasis and proteolysis activation markers. The influence of twice freezing and thawing on clotting factors V, VIII, and XI was also examined. RESULTS: AP contains substantially greater activities of factor (F) V, FVIII, F IX, and FXI than RP frozen within 3 hours after WB donation. Prolonged holding of RP at 20 degrees C for more than 15 hours caused an additional reduction in FVIII, FXI, and protein S activities. Significantly greater levels of prothrombin fragments 1 and 2, platelet factor 4, and neutrophil elastase were found in RP compared with AP. IgG was lower in AP compared with RP. Twice freezing and thawing caused a marked drop in FV, FVIII, and FXI activity. CONCLUSION: Higher FVIII and F IX potencies in AP compared with RP can be expected to result in greater yields when used for purification of these clotting factors. AP is presumably more efficient than RP for treating coagulopathies. RP, however, may contain higher IgG levels than AP. PMID- 15752163 TI - Monocyte enrichment of mononuclear apheresis preparations with a multistep back flush procedure on a cord blood filter. AB - INTRODUCTION: Monocytes or mononuclear cells have been investigated for the treatment of chronic wounds and spinal cord injuries, as well as serve as a source for dendritic or endothelial cell culture. Because these cells may have clinical benefit yet no rapid and inexpensive closed system for monocyte purification is commercially available, a method was investigated to enrich monocytes from mononuclear apheresis units using a cord blood filter. METHODS: A 4-step method for monocyte enrichment was developed which involved 1) filtering a mononuclear apheresis unit through a cord blood filter, 2) chasing with medium to remove non-adherent residual cells and plasma, 3) back-flushing under low shear conditions to remove loosely adherent lymphocytes, and 4) back-flushing under high shear conditions to collect a fraction enriched in monocytes. Apheresis units and enriched monocyte preparations were characterized by cell count and differential, filter-isolated preparations were cryopreserved, and thawed preparations were assayed for viability, and phagocytosis. Enriched monocyte preparations were also assayed for inflammatory cytokines secretion and secretion of prostaglandin E2 during short-term culture. RESULTS: Monocytes were viable, capable of phagocytosis, and enriched using the multistep filter elution technique to represent 42 +/- 13-percent of white cells in the final preparation. Fifty-three-percent of monocytes were recovered in the final preparation, while total cell counts of red cells, platelets, neutrophils and lymphocytes were reduced to 3.0, 3.0, 4.5 and 16-percent, respectively, from levels present in mononuclear apheresis units. Filter enriched monocyte preparations secreted IL-8, IL-6, MCP-1, and MIP-1alpha, during short term culture. CONCLUSION: The use of a multi-step back flush procedure with a cord blood filter resulted in rapid enrichment of viable and functional monocytes from mononuclear apheresis units with significant reduction of contaminating platelets and red cells. PMID- 15752164 TI - The specific power of parentage exclusion in a child's blood relatives. AB - BACKGROUND: The impersonation of parent and child by two other blood relatives is an important problem in parentage analysis involving potential immigrants. STUDY DESIGN AND METHODS: A statistic (AR) is proposed, based on the specific power of exclusion of paternity, which describes the ability of a child's test results to demonstrate evidence of nonparentage under the hypothesis that an ostensible parent is actually an older sibling. A case illustrates the value of A(R): a woman and her two alleged children were typed at 3 variable number of tandem repeat (VNTR) loci after 18 short tandem repeat (STR) loci initially showed strong evidence of the woman's maternity of one child and her exclusion from parentage of the second. AR and 1 - AR were calculated from the STR types of the first child. RESULTS: The woman was excluded from maternity of both children with the additional VNTR tests. Given the 18 STR test findings of the first child, the probability was 12 percent that there would be no inconsistencies with parentage in a sibling pretending to be a parent. CONCLUSION: The value 1 - AR, siblings not excluded from parentage, explains how a seemingly large number of examined loci can fail to reveal even one genetic inconsistency if two siblings have posed as parent and child. Approximately 25 STR loci appear necessary to achieve 95 percent confidence of detecting at least one genetic inconsistency indicative of nonparentage. PMID- 15752165 TI - Mononuclear cell variability and recruitment in non-cytokine-stimulated donors after serial 10-liter leukapheresis procedures. AB - BACKGROUND: We introduced monitoring of mononuclear cell (MNC) counts to obtain enhanced donor control and a stable quality of MNC products, because there are limited data available about blood donors after serial leukapheresis (LP) procedures. STUDY DESIGN AND METHODS: In a prospective paired study, 13 male healthy blood donors underwent 10-L LP procedures performed on two apheresis devices by use of two MNC program settings (COBE Spectra, Gambro BCT, SF 250 vs. SF 500; and AS.TEC 204, Fresenius Hemocare, CP 129 vs. CP 194). Donors' pre- and postdonation MNC counts were analyzed by fluorescence-activated cell sorting. RESULTS: After each 10-L LP procedure, a transient decline (p < 0.05) of CD14+ monocyte and platelet counts appeared in donors. Loss of donors' CD3+ T cells, CD19+ B cells, and CD16+56+ natural killer (NK) cells during MNC collection was partly compensated by cell recruitment. The MNC recruitment factor (RF) seems to be higher with high-yield MNC program settings. Negative correlations (p < 0.01) were noticed between predonation counts and RFs of CD3+ T cells and CD16+56+ NK cells. Four serial 10-L LP procedures did not result in long lasting MNC depletion for donors. CONCLUSION: MNC recruitment seems to depend on MNC program settings and collected cell yields. Low MNC counts could result in high cell recruitment that may contribute to stable collection results to some degree. Nevertheless, there seems to be a considerable individual variation of MNC recruitment in donors that should be investigated in more detail. PMID- 15752167 TI - Risk of Rh(D) alloimmunization after transfusion of platelets from D+ donors to D recipients. PMID- 15752168 TI - ABO discrepancy by usurpation of identity. PMID- 15752169 TI - Selecting cord blood units for storage by CD34+ cell counts. PMID- 15752172 TI - Potential transmission of West Nile virus in the British Isles: an ecological review of candidate mosquito bridge vectors. AB - West Nile virus (WNV) transmitted by mosquitoes (Diptera: Culicidae) infects various vertebrates, being pathogenic for birds, horses and humans. After its discovery in tropical Africa, sporadic outbreaks of WNV occurred during recent decades in Eurasia, but not the British Isles. WNV reached New York in 1999 and spread to California by 2003, causing widespread outbreaks of West Nile encephalitis across North America, transmitted by many species of mosquitoes, mainly Culex spp. The periodic reappearance of WNV in parts of continental Europe (from southern France to Romania) gives rise to concern over the possibility of WNV invading the British Isles. The British Isles have about 30 endemic mosquito species, several with seasonal abundance and other eco-behavioural characteristics predisposing them to serve as potential WNV bridge vectors from birds to humans. These include: the predominantly ornithophilic Culex pipiens L. and its anthropophilic biotype molestus Forskal; tree-hole adapted Anopheles plumbeus Stephens; saltmarsh-adapted Ochlerotatus caspius Pallas, Oc. detritus Haliday and Oc. dorsalis (Meigen); Coquillettidia richiardii Ficalbi, Culiseta annulata Schrank and Cs. morsitans (Theobald) from vegetated freshwater pools; Aedes cinereus Meigen, Oc. cantans Meigen and Oc. punctor Kirby from seasonal woodland pools. Those underlined have been found carrying WNV in other countries (12 species), including the rarer British species Aedes vexans (Meigen), Culex europaeus Ramos et al., Cx. modestus Ficalbi and Oc. sticticus (Meigen) as well as the Anopheles maculipennis Meigen complex (mainly An. atroparvus van Thiel and An. messeae Falleroni in Britain). Those implicated as key vectors of WNV in Europe are printed bold (four species). So far there is no proof of any arbovirus transmission by mosquitoes in the British Isles, although antibodies to Sindbis, Tahyna, Usutu and West Nile viruses have been detected in British birds. Neighbouring European countries have enzootic WNV and human infections transmitted by mosquito species that are present in the British Isles. However, except for localized urban infestations of Cx. pipiens biotype molestus that can be readily eliminated, there appear to be few situations in the British Isles where humans and livestock are exposed to sustained risks of exposure to potential WNV vectors. Monitoring of mosquitoes and arbovirus surveillance are required to guard the British Isles against WNV outbreaks and introduction of more anthropophilic mosquitoes such as Stegomyia albopicta (Skuse) and Ochlerotatus japonicus (Theobald) that have recently invaded Europe, since they transmit arboviruses elsewhere. PMID- 15752173 TI - In vitro rearing of the screwworm fly Wohlfahrtia magnifica. AB - Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae) is the main agent of traumatic myiasis in many European, African and Asian countries. Although it can be reared in vivo without technical difficulty, such rearing presents ethical problems. Studies were therefore made of in vitro rearing to facilitate development of laboratory colonies that could be used in a wide range of biological, physiological and applied studies of W. magnifica, particularly in the long period of the year when natural populations of the fly are unavailable for study. Parental colonies of W. magnifica were established from larvae collected from natural infestations of sheep and cattle in central Hungary. First stage larvae were harvested from gravid females and were reared in groups of 5-20 on one of six artificial diets. The diets were based on various combinations of five to seven of eight ingredients: water, agar, blood (heparinized or dried), ground meat, egg yolk, low-fat milk powder, yeast and 10% formol. The larvae were incubated on the diets at 37 degrees C. There was no mortality of first stage larvae, which appeared to feed together in foci, in a natural manner. However, during the second stage, and especially after renewal of diet associated with disturbance of the larvae, many larvae began to disperse, crawling over the surface of the media and feeding less intensively. Mortality of larvae during all larval stadia was 64-98%, compared to 33% in batches of third stage larvae collected from natural infestations. The mean weights of puparia from artificial diets ranged from 38.7 to 59.3 mg, compared to 92.2 mg of puparia from larvae collected from natural infestations. There was a high mortality in the pupal stage, from 61 to 100%. Only a maximum of 6% of first stage larvae were successfully reared to the adult stage. Further studies are needed to identify factors present or absent in the diets that contributed to the present poor development of W. magnifica in vitro. PMID- 15752174 TI - Malaria transmission dynamics in central Cote d'Ivoire: the influence of changing patterns of irrigated rice agriculture. AB - The dynamics of malaria transmission was studied comparatively in the villages of Zatta and Tiemelekro, central Cote d'Ivoire, from February 2002 to August 2003. Prominent agroecosystems in these villages are irrigated rice growing and vegetable farming, respectively. Mosquitoes (Diptera: Culicidae) were collected on human bait at night and by pyrethrum knock-down spray sheet collections at four randomly selected sentinel sites in each village. In 2002, for a total of 96 man-nights per village, 7716 mosquitoes were collected in Zatta and 3308 in Tiemelekro. In 2003, with half the sampling effort, 859 and 2056 mosquitoes were collected in Zatta and Tiemelekro, respectively. Anopheles gambiae Giles s.l. was the predominant mosquito and the key malaria vector throughout, followed by An. funestus Giles. Anthropophily among adult female Anopheles exceeded 95% in both villages. Comparison between years revealed that the biting rate of An. gambiae s.l. in Zatta decreased several-fold from 49.3 bites per person per night (b/p/n) in 2002 to 7.9 b/p/n in 2003 (likelihood ratio test (LRT) = 1072.66; P < 0.001). Although the biting rate remained fairly constant in Tiemelekro, the difference between years was significant (16.1 vs. 18.2 b/p/n; LRT = 148.06; P < 0.001). These observations were paralleled by a marked decrease in the infective rate of An. gambiae s.l. in Zatta (4.6-1.2%), and an increase in Tiemelekro (3.1-7.6%). Meanwhile, the entomological inoculation rate of An. gambiae s.l. decreased 21 fold in Zatta, from 789 to 38 infective bites per person per year (ib/p/y), whereas it remained high in Tiemelekro (233 vs. 342 ib/p/y). The interruption of irrigated rice growing in Zatta in 2003, consequential to a farmers' conflict over land, might be the underlying cause for the significant reduction in malaria transmission, whereas more stable conditions occurred in Tiemelekro. PMID- 15752175 TI - Micro-geographical variation among male populations of the sandfly, Lutzomyia (Nyssomyia) intermedia, from an endemic area of American cutaneous leishmaniasis in the state of Rio de Janeiro, Brazil. AB - The genetic relationships among male Lutzomyia (Nyssomyia) intermedia (Lutz & Neiva) (Diptera: Psychodidae) from three populations from the same endemic area of American cutaneous leishmaniasis (ACL) in the state of Rio de Janeiro, Brazil, were compared. The sandflies were collected in three ecologically different habitats: domestic, extra-domestic and sylvatic over a total range of 800 m. Three molecular markers were employed to assess population variation. Based on MLEE markers, it could not be concluded that the three populations do not belong to the same gene pool (F(st) = 0.005). No within-population departure from Hardy Weinberg equilibrium was detected (P < 0.05) and they presented the same level of gene variation. The number of migrants (Nm) indicated that at least 50 individuals per generation migrated between the three habitats. RAPD-PCR markers revealed that, except for the primer five, all were polymorphic. Phenetic analysis of the genotypes showed the presence of two principal clusters corresponding to: (1) domestic plus extra-domestic and (2) sylvatic. Unique genotypes were observed in each population. The sylvatic population was the most polymorphic, showing the largest number of genotypes and low level of similarity between them. Three mtDNA gene markers were studied by SSCP analysis. The most frequent haplotype for each marker ranged in frequency from 60 to 87% and individuals with unique haplotypes varied from 1 to 5%. Interestingly, the SSCP analysis showed a low level of polymorphism within populations. The disagreement between the different molecular markers observed and the hypothesis that L. intermedia could be participating in the transmission cycle of Leishmania (Viannia) braziliensis in environments ranging from the interior of human dwellings to the forest, are discussed. PMID- 15752176 TI - North American face flies Old World origins: mitochondrial evidence. AB - Analysis of a 513 base sequence of mitochondrial cytochrome oxidase I gene in Musca autumnalis De Geer (Diptera: Muscidae) from the U.S.A., England, Russia and Kazakhstan confirms that North American flies originated in Western Europe. Flies from the U.S.A., England and southern Russia shared most of their mitochondrial diversities, but face flies from Kazakhstan were substantially dissimilar, suggesting highly restricted gene flow and a species complex within the Palearctic. PMID- 15752177 TI - Spatial diversity in mitochondrial cytochrome c oxidase in house flies. AB - DNA sequence analysis at mitochondrial gene COI was surveyed in 293 house flies, Musca domestica Linneaus (Diptera: Muscidae), in 29 populations from North, Central and South America, Europe, Asia, Africa, and the Western Pacific. Nei's gene diversity index (H(S)) was 0.47, the chance that two randomly chosen flies have different COI haplotypes. Haplotype diversity was greater in the Old World (H(S) = 0.58) than the New World (H(S) = 0.31). The hierarchical partition of the total diversity indicated substantial differentiation at all levels (G(ST) = 0.30), and highly structured populations. All pairwise estimates of gene flow between zoogeographical regions were less than 0.70 reproducing females per generation. The results are compared to those of a similar study based on the single-strand conformation polymorphism method. Probable colonization scenarios for house flies into the New World are discussed and it is concluded that house flies are a recent addition to the fauna of the Western Hemisphere. PMID- 15752178 TI - Filarial susceptibility and effects of Wolbachia in Aedes pseudoscutellaris mosquitoes. AB - The mosquito Aedes pseudoscutellaris (Theobald), a member of the Aedes (Stegomyia) scutellaris complex (Diptera: Culicidae), is an important vector of subperiodic Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae), causing human lymphatic filariasis, on South Pacific islands. Maternal inheritance of filarial susceptibility in the complex has previously been asserted, and larval tetracycline treatment reduced susceptibility; the maternally inherited Wolbachia in these mosquitoes were suggested to be responsible. To investigate the relationship of these two factors, we eliminated Wolbachia from a strain of Ae. pseudoscutellaris by tetracycline treatment, and tested filarial susceptibility of the adult female mosquitoes using Brugia pahangi (Edeson & Buckley). Filarial susceptibility was not significantly different in Wolbachia-free and infected lines of Ae. pseudoscutellaris, suggesting that the Wolbachia in these mosquitoes do not influence vector competence. Crosses between Wolbachia-infected males and uninfected females of Ae. pseudoscutellaris showed cytoplasmic incompatibility (CI), i.e. no eggs hatched, unaffected by larval crowding or restricted nutrient availability, whereas these factors are known to affect CI in Drosophila simulans. Reciprocal crosses between Ae. pseudoscutellaris and Ae. katherinensis Woodhill produced no progeny, even when both parents were Wolbachia-free, suggesting that nuclear factors are responsible for this interspecific sterility. PMID- 15752179 TI - Population differentiation and Wolbachia phylogeny in mosquitoes of the Aedes scutellaris group. AB - Mosquito species of the Aedes (Stegomyia) scutellaris (Walker) group (Diptera: Culicidae) are distributed across many islands of the South Pacific and include major regional vectors of filariasis, such as Aedes polynesiensis (Marks). Analysis of populations of Ae. polynesiensis at the extremes of its range, from Fiji and from Moorea, French Polynesia, using the rDNA ITS2 (internal transcribed spacer 2) region and six microsatellite markers showed considerable genetic differentiation between them (F(ST) = 0.298-0.357). Phylogenetic analysis of the Wolbachia endosymbionts in three members of the complex revealed that based on the wsp gene they are all very similar and belong to the Mel subgroup of the A clade, closely related to the Wolbachia strain present in the gall wasp Callyrhytis glandium (Giraud) (Hymenoptera: Cynipidae). By contrast they are only distantly related to the A-clade Wolbachia in Aedes albopictus (Skuse), a species closely allied to the Ae. scutellaris group. There was very low differentiation between the Wolbachia in the Moorea and Fiji populations of Ae. polynesiensis. PMID- 15752180 TI - Multi-country field trials comparing wash-resistance of PermaNet and conventional insecticide-treated nets against anopheline and culicine mosquitoes. AB - Insecticide-treated bednets (ITNs) are commonly used as a means of personal protection from malaria transmission by anopheline mosquitoes (Diptera: Culicidae). Long-lasting insecticidal nets (LLINs) have special treatments intended to remain effective after many washes. The present trials assessed the efficacy and wash-resistance of several production batches of PermaNet (polyester net coated with polymer resin containing pyrethroid insecticide deltamethrin 55 mg ai/m2) against malaria vectors in Pakistan, Iran and Tanzania compared to ITNs conventionally treated with alphacypermethrin 15 or 20 mg ai/m2, or deltamethrin 25 or 50 mg ai/m2. Insecticidal efficacy of the nets before and after repeated washing (using W.H.O. recommended and traditional local washing procedures) was monitored through contact bioassays with Anopheles and by experimental hut and outdoor platform tests. Local washing regimes gradually reduced the insecticidal efficacy of conventionally treated nets, but they were not exhausted, even after 21 washes. Using a more rigorous laboratory washing method, insecticide was more readily stripped from conventionally treated nets. PermaNet retained high efficacy after 21 washes, giving more than 97% mortality of Anopheles in contact bioassays with 3-min exposure. Using the more sensitive bioassay criterion of 'median time to knockdown', PermaNet showed no loss of insecticidal activity against Anopheles after washing repeatedly in 2 out of 6 trials; whereas in a further three trials knockdown activity of PermaNet and conventional ITNs declined at comparable rates. Higher mortality levels of Anopheles in contact bioassays did not always translate to superiority in experimental hut or enclosed platform trials. In only one of four comparative field trials did PermaNet out perform conventional ITNs after washing: this was in the trial of PermaNet 2.0- the product with improved quality assurance. Because PermaNet and conventionally treated nets were both quite tolerant of local washing procedures, it is important in field trials to compare LLINs with conventional ITNs washed an equivalent number of times. Our comparison of PermaNet 2.0 against conventionally treated deltamethrin nets (CTDN) in Pakistan demonstrated superior performance of the LLIN after 20 washes in phase I and phase II bioassays, and this was corroborated by chemical assays of residual deltamethrin. Although PermaNet 2.0 has received WHOPES interim recommendation for malaria control purposes, its performance should be monitored in everyday use throughout its lifespan in various cultural settings to assess its durability and long-term effectiveness for malaria prevention and control. As many millions of conventionally treated nets are already in routine use, and these will require regular re-treatment, programme strategies should be careful to preserve the effectiveness of ITNS before and after establishing the reliability of LLINs in long-term use. PMID- 15752181 TI - A double-blinded, placebo-controlled trial of garlic as a mosquito repellant: a preliminary study. AB - The hypothesis that the ingestion of garlic provides protection against bloodsucking pests such as mosquitoes was investigated using a randomized, double blinded, placebo-controlled crossover study. Subjects were asked to consume either garlic (one visit) or a placebo (the other visit). They were then exposed to laboratory-reared Aedes aegypti (Linnaeus) (Diptera: Culicidae). The numbers of mosquitoes that did not feed on the subjects, the number of mosquito bites, the weights of the mosquitoes after feeding and the amounts of blood ingested were determined. The data did not provide evidence of significant systemic mosquito repellence. A limitation of the study is that more prolonged ingestion of garlic may be needed to accomplish repellence. PMID- 15752182 TI - Borreliacidal activity of saliva of the tick Amblyomma americanum. AB - Amblyomma americanum (Linneaus) (Acari: Ixodidae), an important tick vector of human and animal disease, is not a competent vector of the bacterial agent of Lyme disease, Borrelia burgdorferi, although its range overlaps the geographical distribution of Lyme disease within the United States. A possible mechanism that could prevent acquisition of B. burgdorferi spirochetes from infected hosts is the toxic effect of A. americanum saliva on B. burgdorferi. The data presented here indicate that after 24 and 48 h of exposure to A. americanum saliva, significantly fewer B. burgdorferi were alive compared to treatment controls as assessed by spirochete motility under dark-field microscopy and resistance to the dead stain, propidium iodide. After 48 h, fewer than 13% of saliva-exposed B. burgdorferi were alive. In contrast, significantly more B. burgdorferi exposed to Ixodes scapularis (Acari: Ixodidae) saliva survived after 24 or 48 h compared to A. americanum saliva or treatment controls. PMID- 15752183 TI - Morphological variability and genetic identity in Rhinoestrus spp. causing horse nasal myiasis. AB - Larvae of Rhinoestrus purpureus (Brauer) and Rhinoestrus usbekistanicus Gan (Diptera: Oestridae) cause nasal myiases of equids. During a recent epidemiological survey in southern Italy some morphological and taxonomical doubts arose concerning the identification of Rhinoestrus third stage larvae on the basis of the features of the posterior spiracles and the distribution of dorsal spines on the third segment. Four different morphotypes were retrieved: R. usbekistanicus-like, R. purpureus-like and two morphotypes with shared features. The genes encoding for the mitochondrial cytochrome oxidase I (COI) and for the ribosomal subunits 16S and 28S of the four morphotypes of Rhinoestrus were investigated to determine whether they belonged to a single taxon or they displayed genetic differences indicative of more than one species. The three genes showed a very low level of sequence variation (COI 0-0.43%, 16S 0-1.45%, 28S 0-0.23%) falling within the intraspecific ranges previously described for Oestridae species. Finally, the peritreme features and the spinulation of the third segment of the four morphotypes examined could not be used to differentiate the two species. PMID- 15752184 TI - Repellency of deet and SS220 applied to skin involves olfactory sensing by two species of ticks. AB - Responses of host-seeking nymphs of the blacklegged tick, Ixodes scapularis Say and lone star tick, Amblyomma americanum (Linnaeus) (Acari: Ixodidae) to the repellents N,N-diethyl-3-methylbenzamide (deet) and (1S, 2'S)-2-methylpiperidinyl 3-cyclohexene-1-carboxamide (SS220) were studied using fingertip laboratory bioassays. Ethanol solutions of both compounds applied to the skin strongly repelled both species of ticks at 0.8 and 1.6 micromole of compound/cm2 skin. The ticks were also repelled when two layers of organdie cloth covered the portion of a finger treated with either deet or SS220. Gas chromatographic analyses of the outer layer of cloth that had covered skin treated with 1.6 micromole compound/cm2 skin revealed only 0.1 nmole SS220/cm2 cloth and 2.8 nmole deet/cm2 cloth. However, in bioassays in which a single layer of cloth was treated with a dose of deet or SS220 equivalent to the amount found in the outer layer of cloth, ticks were not repelled. Results unequivocally demonstrated that these ticks responded to the repellents in the vapour phase when repellent treated skin was covered with cloth to obviate tactile contact with them, and made it clear that the ticks detect the repellents by olfactory sensing. Heretofore, the mode of action of deet and SS220 was unclear. PMID- 15752185 TI - Sand fly species of Sanliurfa province in Turkey. AB - The species composition and seasonal abundance of sand flies (Diptera: Phlebotominae) were studied in the years 2000-2002 in the Sanliurfa region, which is the largest focus of cutaneous leishmaniasis in south-eastern Turkey. Sixteen species were identified among 29 771 specimens collected at 17 different sites by light traps, sticky papers and aspirators. The most common species were Phlebotomus papatasi (Scopoli) (45.4%), P. perfiliewi Parrot (21.9%), and P. sergenti Parrot (19.4%). The other species found were P. major Adler & Theodor (3%), P. neglectus Leger & Pesson (2.2%), P. brevis Theodor & Mesghali (2%), P. alexandri Sinton (1.9%), P. galilaeus Theodor (1.6%), P. halepensis Theodor (0.84%), Sergentomyia adleri Theodor (0.78%), S. dentata Sinton (0.49%), S. minuta Rondani (0.42%), S. theodori Parrot (0.16%), P. kazeruni Theodor & Mesghali (0.001%) and P. mascitti Grassi (0.001%) and one unidentified Phlebotomus species. Among these species P. galilaeus, S. minuta and S. dentata are the first records for this area. All species showed seasonal fluctuations, with the period of highest abundance between May and October. PMID- 15752186 TI - Distribution of phlebotomine sandflies, of the subgenus Larroussius, in Morocco. AB - Adult sandflies (Diptera: Psychodidae) of the genus Phlebotomus and of the subgenus Larroussius were collected with sticky traps from thirty-four localities in central and south-western Morocco. Four Larroussius species were present in this area, Phlebotomus perniciosus Newstead, being the most widespread and abundant species particularly in the mountainous area, was sympatric with the closely related species Phlebotomus longicuspis Nitzulescu. Only one male of P. perniciosus displayed the bifid copulatory valves characteristic of the species. All specimens showed copulatory valves curved at their apex. The number of coxite hairs is a taxonomical tool that differentiates this morph of P. perniciosus from P. longicuspis. Phlebotomus ariasi Tonnoir was collected from six mountainous stations. Phlebotomus mariae Rioux et al. was scarce; only five males were collected and were often associated with P. ariasi. PMID- 15752187 TI - Changes in the haemocyte population of the mosquito, Culex quinquefasciatus, following infection with the filarial parasite, Wuchereria bancrofti. AB - The mosquito Culex quinquefasciatus Say (Diptera: Culicidae) is the vector of the filarial parasite Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae), which causes human bancroftian filariasis. Information on the mosquito humoral response against the filarial parasite during the process of its infection and development is important, as it decides the vector competence of the mosquito. Visible changes in the haemocyte population of mosquito, if any, will be an indicator of the possible humoral factors. The present study was aimed at investigating changes in the populations of various types of haemocytes of Cx. quinquefasciatus following infection with W. bancrofti. On day 2 post-feeding on microfilaraemic blood, the haemolymph perfusate of infected mosquitoes with L1 stage of the parasite showed 44.1% granulocytes, 42% prohaemocytes and 13.9% plasmatocytes, whereas that of the control mosquitoes fed on amicrofilaraemic blood showed 63.4% plasmatocytes, 22.2% prohaemocytes and 14.4% granulocytes. Differences in the population numbers of haemocyte types between the infected and control were significant (P > 0.05). However, the mosquitoes examined on day 6 post-feeding, when the parasite was in L2 stage, did not show any such changes. But, similar changes reappeared on day 12 in mosquitoes with L3 stage of the parasite. The observed haemocyte population changes indicate the possibility of some amount of humoral immune response, through the production of certain immune molecules, in Cx. quinquefasciatus infected with W. bancrofti. The nature and exact role of such a response on the filarial parasite development need further investigation. PMID- 15752188 TI - A new release on life: emerging concepts in proteolysis and parasite invasion. AB - Cell invasion by apicomplexan pathogens such as the malaria parasite and Toxoplasma is accompanied by extensive proteolysis of zoite surface proteins (ZSPs) required for attachment and penetration. Although there is still little known about the proteases involved, a conceptual framework is emerging for the roles of proteolysis in cell invasion. Primary processing of ZSPs, which includes the trimming of terminal peptides or segmentation into multiple fragments, is proposed to activate these adhesive ligands for tight binding to host receptors. Secondary processing, which occurs during penetration, results in the shedding of ZSPs by one of two mechanistically distinct ways, shaving or capping. Resident surface proteins are typically shaved from the surface whereas adhesive ligands mobilized from intracellular secretory vesicles are capped to the posterior end of the parasite before being shed during the final steps of penetration. Intriguingly, recent studies have revealed that ZSPs can be released either by being cleaved adjacent to the membrane anchor or actually within the membrane itself. Mounting evidence suggests that intramembrane cleavage is catalysed by one or more integral membrane serine proteases of the Rhomboid family and we propose that several malaria adhesive ligands may be potential substrates for these enzymes. We also discuss the evidence that the key reason for ZSP shedding during invasion is to break the connection between parasite surface ligands and host receptors. The sequential proteolytic events associated with invasion by pathogenic protozoa may represent vulnerable pathways for the future development of synergistic anti-protozoal therapies. PMID- 15752189 TI - Maturation of the Escherichia coli divisome occurs in two steps. AB - Cell division proteins FtsZ (FtsA, ZipA, ZapA), FtsE/X, FtsK, FtsQ, FtsL/B, FtsW, PBP3, FtsN and AmiC localize at mid cell in Escherichia coli in an interdependent order as listed. To investigate whether this reflects a time dependent maturation of the divisome, the average cell age at which FtsZ, FtsQ, FtsW, PBP3 and FtsN arrive at their destination was determined by immuno- and GFP-fluorescence microscopy of steady state grown cells at a variety of growth rates. Consistently, a time delay of 14-21 min, depending on the growth rate, between Z ring formation and the mid cell recruitment of proteins down stream of FtsK was found. We suggest a two-step model for bacterial division in which the Z-ring is involved in the switch from cylindrical to polar peptidoglycan synthesis, whereas the much later localizing cell division proteins are responsible for the modification of the envelope shape into that of two new poles. PMID- 15752190 TI - A magnesium-dependent mreB null mutant: implications for the role of mreB in Bacillus subtilis. AB - MreB shares a common prokaryotic ancestor with actin and is present in almost all rod-shaped bacteria. MreB proteins have been implicated in a range of important cell processes, including cell morphogenesis, chromosome segregation and cell polarity. The mreB gene frequently lies at the beginning of a cluster of genes, immediately upstream of the conserved mreC and mreD genes. RNA analysis showed that in Bacillus subtilis mreB is co-transcribed with mreC and that these genes form part of an operon under the control of a promoter(s) upstream of mreB. Construction of an in-frame deletion of mreB and its complementation by mreB(+) only, in trans, established that the gene is important for maintenance of cell width and cell viability under normal growth conditions, independent of polar effects on downstream genes. Remarkably, virtually normal growth was restored to the mreB null mutant in the presence of high concentrations of magnesium, especially when high concentrations of the osmoprotectant, sucrose were also present. Under these conditions, cells could be maintained in the complete absence of an mreB gene, with almost normal morphology. No detectable effect on chromosome segregation was evident in the mutant, nor was there an effect on the topology of nascent peptidoglycan insertion. A GFP-MreB fusion was used to look at the localization of MreB in live cells. The pattern of localization was similar to that previously described, but no tight linkage to nucleoid positioning was evident. Propagation of the mreB null mutant in the absence of magnesium and sucrose led to a progressive increase in cell width, culminating in cell lysis. Cell division was also perturbed but this effect may be secondary to the disturbance in cell width. These results suggest that the major role of MreB in B. subtilis lies in the control of cell diameter. PMID- 15752191 TI - Structural and functional studies of the enteropathogenic Escherichia coli type III needle complex protein EscJ. AB - The type III secretion system (TTSS) is a macromolecular structure that spans the cell wall of Gram-negative bacterial pathogens, enabling delivery of virulence effector proteins directly to the membranes and cytosol of host eukaryotic cells. TTSS consists of a conserved needle complex (NC) that is composed of sets of inner and outer membranes rings connected by a periplasmic rod. Enteropathogenic Escherichia coli (EPEC) is an extracellular diarrhoeagenic pathogen that uses TTSS to induce actin polymerization and colonizes the intestinal epithelium. In EPEC, EscJ is predicted to be targeted to the periplasm, in a sec-dependent manner, and to bridge the TTSS membrane-associated rings. In this study we determined the global fold of EscJ using Nuclear Magnetic Resonance spectroscopy. We show that EscJ comprises two subdomains (D1 - amino acid residues 1-55 in the mature protein, and D2 - amino acid residues 90-170), each comprising a three stranded beta-sheet flanked by two alpha-helices. A flexible region (residues 60 85) couples the structured regions D1 and D2. Periplasmic overexpression of EscJ(D1) and EscJ(D2) in a single escJ mutant bacterium failed to restore protein secretion activity, suggesting that the flexible linker is essential for the rod function. In contrast, periplasmic overexpression of EscJ(D1) and EscJ(D2) in the same wild-type bacterium had a dominant-negative phenotype suggesting defective assembly of the TTSS and protein translocation. PMID- 15752192 TI - Genetic analysis of mch mutants in two Methanosarcina species demonstrates multiple roles for the methanopterin-dependent C-1 oxidation/reduction pathway and differences in H(2) metabolism between closely related species. AB - A mutation in the mch gene, encoding the enzyme 5,10-methenyl tetrahydromethanopterin (H(4)MPT) cyclohydrolase, was constructed in vitro and recombined onto the chromosome of the methanogenic archaeon Methanosarcina barkeri. The resulting mutant does not grow in media using H(2)/CO(2), methanol, or acetate as carbon and energy sources, but does grow in media with methanol/H(2)/CO(2), demonstrating its ability to utilize H(2) as a source of electrons for reduction of methyl groups. Cell suspension experiments showed that methanogenesis from methanol or from H(2)/CO(2) is blocked in the mutant, explaining the lack of growth on these substrates. The corresponding mutation in Methanosarcina acetivorans C2A, which cannot grow on H(2)/CO(2), could not be made in wild-type strains, but could be made in strains carrying a second copy of mch, suggesting that M. acetivorans is incapable of methyl group reduction using H(2). M. acetivorans mch mutants could also be constructed in strains carrying the M. barkeri ech hydrogenase operon, suggesting that the block in the methyl reduction pathway is at the level of H(2) oxidation. Interestingly, the ech dependent methyl reduction pathway of M. acetivorans involves an electron transport chain distinct from that used by M. barkeri, because M. barkeri ech mutants remain capable of H(2)-dependent methyl reduction. PMID- 15752193 TI - MpcT is the transducer for membrane potential changes in Halobacterium salinarum. AB - In Halobacterium salinarum mutants containing either of the light-driven ion pumps bacteriorhodopsin (H(+)) or halorhodopsin (Cl(-)) as their only retinal protein, a decrease of irradiance in the absence of respiration causes a phototactic response. The conversion of the causal event, a decrease of proton motive force across the cell membrane, into a reversal of flagellar motor rotational direction was expected to involve a transducer. Via deletion analysis of all 18 known and putative halobacterial transducer (htr) genes, we found that Htr14, a methylatable membrane-bound transducer lacking an extracellular domain, mediates the biological response, which includes adaptive methylation. Based on a minimal stimulus length of 200 ms and the determined cytoplasmic buffering capacity, we conclude that the change in the membrane potential (DeltaPsi), and not that of the internal pH, is the signal-generating event. Htr14 was therefore renamed to Membrane potential change Transducer, or MpcT. It is the first transducer for which the causative stimulus could be narrowed to a change in DeltaPsi, as opposed to a change in pH or cellular redox state. PMID- 15752194 TI - Functional analysis of the Campylobacter jejuni N-linked protein glycosylation pathway. AB - We describe in this report the characterization of the recently discovered N linked glycosylation locus of the human bacterial pathogen Campylobacter jejuni, the first such system found in a species from the domain Bacteria. We exploited the ability of this locus to function in Escherichia coli to demonstrate through mutational and structural analyses that variant glycan structures can be transferred onto protein indicating the relaxed specificity of the putative oligosaccharyltransferase PglB. Structural data derived from these variant glycans allowed us to infer the role of five individual glycosyltransferases in the biosynthesis of the N-linked heptasaccharide. Furthermore, we show that C. jejuni- and E. coli-derived pathways can interact in the biosynthesis of N-linked glycoproteins. In particular, the E. coli encoded WecA protein, a UDP-GlcNAc: undecaprenylphosphate GlcNAc-1-phosphate transferase involved in glycolipid biosynthesis, provides for an alternative N-linked heptasaccharide biosynthetic pathway bypassing the requirement for the C. jejuni-derived glycosyltransferase PglC. This is the first experimental evidence that biosynthesis of the N-linked glycan occurs on a lipid-linked precursor prior to transfer onto protein. These findings provide a framework for understanding the process of N-linked protein glycosylation in Bacteria and for devising strategies to exploit this system for glycoengineering. PMID- 15752195 TI - Neisseria gonorrhoeae secretes chromosomal DNA via a novel type IV secretion system. AB - The process of DNA donation for natural transformation of bacteria is poorly understood and has been assumed to involve bacterial cell death. Recently in Neisseria gonorrhoeae we found that mutations in three genes in the gonococcal genetic island (GGI) reduced the ability of a strain to act as a donor in transformation and to release DNA into the culture. To better characterize the GGI and the process of DNA donation, the 57 kb genetic island was cloned, sequenced and subjected to insertional mutagenesis. DNA sequencing revealed that the GGI has characteristics of a horizontally acquired genomic island and encodes homologues of type IV secretion system proteins. The GGI was found to be incorporated near the chromosomal replication terminus at the dif site, a sequence targeted by the site-specific recombinase XerCD. Using a plasmid carrying a small region of the GGI and the associated dif site, we demonstrated that this model island could be integrated at the dif site in strains not carrying the GGI and was spontaneously excised from that site. Also, we were able to delete the entire 57 kb region by transformation with DNA from a strain lacking the GGI. Thus the GGI was likely acquired and integrated into the gonococcal chromosome by site-specific recombination and may be lost by site specific recombination or natural transformation. We made mutations in six putative type IV secretion system genes and assayed these strains for the ability to secrete DNA. Five of the mutations greatly reduced or completely eliminated DNA secretion. Our data indicate that N. gonorrhoeae secretes DNA via a specific process. Donated DNA may be used in natural transformation, contributing to antigenic variation and the spread of antibiotic resistance, and it may modulate the host immune response. PMID- 15752196 TI - Tethering the Z ring to the membrane through a conserved membrane targeting sequence in FtsA. AB - The cytokinetic Z ring is required for bacterial cell division. It consists of polymers of FtsZ, the bacterial ancestor of eukaryotic tubulin, linked to the cytoplasmic membrane. Formation of a Z ring in Escherichia coli occurs as long as one of two proteins, ZipA or FtsA, is present. Both of these proteins bind FtsZ suggesting that they might function to tether FtsZ filaments to the membrane. Although ZipA has a transmembrane domain and therefore can function as a membrane anchor, interaction of FtsA with the membrane has not been explored. In this study we demonstrate that FtsA, which is structurally related to eukaryotic actin, has a conserved C-terminal amphipathic helix that is essential for FtsA function. It is required to target FtsA to the membrane and subsequently to the Z ring. As FtsA is much more widely conserved in bacteria than ZipA, it is likely that FtsA serves as the principal membrane anchor for the Z ring. PMID- 15752197 TI - Qri2/Nse4, a component of the essential Smc5/6 DNA repair complex. AB - We demonstrate a role for Qri2 in the essential DNA repair function of the Smc5/6 complex in Saccharomyces cerevisiae. We generated temperature-sensitive (ts) mutants in QRI2 and characterized their properties. The mutants arrest after S phase and prior to mitosis. Furthermore, the arrest is dependant on the Rad24 checkpoint, and is also accompanied by phosphorylation of the Rad53 checkpoint effector kinase. The mutants also display genome instability and are sensitive to agents that damage DNA. Two-hybrid screens reveal a physical interaction between Qri2 and proteins that are non-Smc elements of the Smc5/6 DNA repair complex, which is why we propose the name NSE4 for the open reading frame previously known as QRI2. A key role for Nse4 in Smc5/6 function is likely, as overexpressing known subunits of the Smc5/6 complex suppresses nse4(ts) cell cycle arrest. The nse4(ts) growth arrest is non-lethal and unlike the catastrophic nuclear fragmentation phenotype of smc6(ts) mutants, the nucleus remains intact; replicative intermediates and sheared DNA are not detected. This could imply a role for Nse4 in maintenance of higher order chromosome structure. PMID- 15752198 TI - Mutant forms of the Escherichia colibeta sliding clamp that distinguish between its roles in replication and DNA polymerase V-dependent translesion DNA synthesis. AB - The Escherichia colibeta sliding clamp is proposed to play an important role in regulating DNA polymerase traffic at the replication fork. As part of an ongoing effort to understand how organisms manage the actions of their multiple DNA polymerases, we examined the ability of several mutant forms of the beta clamp to function in DNA polymerase V- (pol V-) dependent translesion DNA synthesis (TLS) in vivo. Our results indicate that a dnaN159 strain, which expresses a temperature sensitive form of the beta clamp, was impaired for pol V-dependent TLS at the permissive temperature of 37 degrees C. This defect was complemented by a plasmid that expressed near-physiological levels of the wild-type clamp. Using a dnaN159 mutant strain, together with various plasmids expressing mutant forms of the clamp, we determined that residues H148 through R152, which comprise a portion of a solvent exposed loop, as well as position P363, which is located in the C-terminal tail of the beta clamp, are critically important for pol V dependent TLS in vivo. In contrast, these same residues appear to be less critical for pol III-dependent replication. Taken together, these findings indicate that: (i) the beta clamp plays an essential role in pol V-dependent TLS in vivo and (ii) pol III and pol V interact with non-identical surfaces of the beta clamp. PMID- 15752199 TI - Subcellular localization of a sporulation membrane protein is achieved through a network of interactions along and across the septum. AB - During the process of spore formation in Bacillus subtilis many membrane proteins localize to the sporulation septum where they play key roles in morphogenesis and cell-cell signalling. However, the mechanism by which these proteins are anchored at this site is not understood. In this report we have defined the localization requirements for the mother-cell membrane protein SpoIVFA, which anchors a signalling complex in the septal membrane on the mother cell side. We have identified five proteins (SpoIID, SpoIIP, SpoIIM, BofA and SpoIIIAH) synthesized in the mother cell under the control of sigma(E) and one protein (SpoIIQ) synthesized in the forespore under the control of sigma(F) that are all required for the proper localization of SpoIVFA. Surprisingly, these proteins appear to have complementary and overlapping anchoring roles suggesting that SpoIVFA is localized in the septal membrane through a web of protein interactions. Furthermore, we demonstrate a direct biochemical interaction between the extracellular domains of two of the proteins required to anchor SpoIVFA: the forespore protein SpoIIQ and the mother-cell protein SpoIIIAH. This result supports the idea that the web of interactions that anchors SpoIVFA is itself held in the septal membrane through a zipper-like interaction across the sporulation septum. Importantly, our results suggest that a second mechanism independent of forespore proteins participates in anchoring SpoIVFA. Finally, we show that the dynamic localization of SpoIIQ in the forespore is impaired in the absence of SpoIVFA but not SpoIIIAH. Thus, a complex web of interactions among mother cell and forespore proteins is responsible for static and dynamic protein localization in both compartments of the sporangium. We envision that this proposed network is involved in anchoring other sporulation proteins in the septum and that protein networks with overlapping anchoring capacity is a feature of protein localization in all bacteria. PMID- 15752201 TI - Evidence for non-enzymatic glycosylation of Escherichia coli chromosomal DNA. AB - We have recently shown that the process of non-enzymatic glycosylation (glycation) takes place in Escherichia coli under physiological conditions and affects both recombinant and endogenous bacterial proteins. In this study, we further demonstrate that E. coli chromosomal DNA is also subjected to glycation under physiological growth conditions. The E. coli DNA accumulates early glycation (Amadori) products as proven by the nitroblue tetrazolium (NBT) reduction assay. It showed also immunoreactivity to a monoclonal antibody raised against N(in)-(carboxymethyl)lysine and fluorescent properties indicative of modifications with advanced glycation end-products. Two types of fluorophores were detected in the E. coli DNA with excitation maxima at 360 nm and 380 nm and emission maxima at 440 nm and 410 nm. Using the NBT reduction assay, fluorescence spectroscopy and enzyme-linked immunosorbent assay we revealed that glycation adducts accumulate in DNA predominantly in the stationary phase of growth, although they could be detected also in exponential-phase cells. Besides on the growth phase, the extent of DNA glycation depends also on the nutrient broth composition being more extensive in rich media. Thiamine was found to inhibit both DNA glycation and spontaneous point mutations as judged by the decreased rate of the argE3 to Arg(+) reversions in the E. coli strain AB1157. PMID- 15752200 TI - Characterization of Cryptococcus neoformans variety gattii SOD2 reveals distinct roles of the two superoxide dismutases in fungal biology and virulence. AB - We studied superoxide dismutases (SODs) in the encapsulated yeast Cryptococcus neoformans (Cn) variety gattii to analyse the role of mitochondrial MnSOD (SOD2) in fungal biology and virulence. SOD2 was cloned from a Cn cosmid library, sod2 mutant and sod2 + SOD2 reconstituted strains were constructed by homologous recombination, and two sod1sod2 double mutants were constructed by replacing SOD2 in the sod1 mutant with the sod2::HYG allele. The SOD2 protein (SOD2p) encoded 225 amino acids, with 36-66% identity with other fungal SOD2ps. SOD2 deletion rendered Cn highly growth-defective at 37 degrees C in 19-20% oxygen (normal air), and this defect was reversed by limiting oxygen to 1.3% as well in the presence of antioxidant, ascorbic acid. The sod2 mutant accumulated significantly more reactive oxygen species (ROS) at 37 degrees C as well at 30 degrees C in the presence of antimycin A, suggesting that SOD2p is the primary defence of Cn against the superoxide anion (O(2) (.-)) in the mitochondria. The sod2 was also highly susceptible to redox-cycling agents, high salt and nutrient limitations. The sod2 mutant was avirulent in intranasally infected mice and markedly attenuated in its virulence in intravenously infected mice. The virulence defect of sod2 mutant appeared related to its growth defects in high oxygen environment, but not resulting from increased sensitivity to oxidative killing by phagocytes. The sod1sod2 double mutants were avirulent in mice. Additionally, sod1sod2 double mutants showed a marked reduction in the activities of other known Cn virulence factors; and they were more susceptible to PMN killing than was the sod2 single mutant. Previously, we reported that the attenuation of sod1 mutant in mice was resulting from enhanced susceptibility to phagocyte killing, combined with a reduction in the activities of a number of virulence factors. Thus, SOD1p and SOD2p play distinct roles in the biology and virulence of Cn var. gattii via independent modes of action. PMID- 15752202 TI - The expanding world of small RNAs in the hyperthermophilic archaeon Sulfolobus solfataricus. AB - Archaeal L7Ae is a multifunctional protein that binds to a distinctive K-turn motif in RNA and is found as a component in the large subunit of the ribosome, and in ribose methylation and pseudouridylation guide RNP particles. A collection of L7Ae-associated small RNAs were isolated from Sulfolobus solfataricus cell extracts and used to construct a cDNA library; 45 distinct cDNA sequences were characterized and divided into six groups. Group 1 contained six RNAs that exhibited the features characteristic of the canonical C/D box archaeal sRNAs, two RNAs that were atypical C/D box sRNAs and one RNA representative of archaeal H/ACA sRNA family. Group 2 contained 13 sense strand RNA sequences that were encoded either within, or overlapping annotated open reading frames (ORFs). Group 3 contained three sequences form intergenic regions. Group 4 contained antisense sequences from within or overlapping sense strand ORFs or antisense sequences to C/D box sRNAs. More than two-thirds of these sequences possessed K-turn motifs. Group 5 contained two sequences corresponding to internal regions of 7S RNA. Group 6 consisted of 11 sequences that were fragments from the 5' or 3' ends of 16S and 23S ribosomal RNA and from seven different tRNAs. Our data suggest that S. solfataricus contains a plethora of small RNAs. Most of these are bound directly by the L7Ae protein; the others may well be part of larger, transiently stable RNP complexes that contain the L7Ae protein as core component. PMID- 15752203 TI - The Mycobacterium tuberculosis iniA gene is essential for activity of an efflux pump that confers drug tolerance to both isoniazid and ethambutol. AB - Little is known about the intracellular events that occur following the initial inhibition of Mycobacterium tuberculosis by the first-line antituberculosis drugs isoniazid (INH) and ethambutol (EMB). Understanding these pathways should provide significant insights into the adaptive strategies M. tuberculosis undertakes to survive antibiotics. We have discovered that the M. tuberculosis iniA gene (Rv 0342) participates in the development of tolerance to both INH and EMB. This gene is strongly induced along with iniB and iniC (Rv 0341 and Rv 0343) by treatment of Mycobacterium bovis BCG or M. tuberculosis with INH or EMB. BCG strains overexpressing M. tuberculosis iniA grew and survived longer than control strains upon exposure to inhibitory concentrations of either INH or EMB. An M. tuberculosis strain containing an iniA deletion showed increased susceptibility to INH. Additional studies showed that overexpression of M. tuberculosis iniA in BCG conferred resistance to ethidium bromide, and the deletion of iniA in M. tuberculosis resulted in increased accumulation of intracellular ethidium bromide. The pump inhibitor reserpine reversed both tolerance to INH and resistance to ethidium bromide in BCG. These results suggest that iniA functions through an MDR-pump like mechanism, although IniA does not appear to directly transport INH from the cell. Analysis of two-dimensional crystals of the IniA protein revealed that this predicted transmembrane protein forms multimeric structures containing a central pore, providing further evidence that iniA is a pump component. Our studies elucidate a potentially unique adaptive pathway in mycobacteria. Drugs designed to inhibit the iniA gene product may shorten the time required to treat tuberculosis and may help prevent the clinical emergence of drug resistance. PMID- 15752204 TI - Unique mechanistic features of post-translational regulation of glutamine synthetase activity in Methanosarcina mazei strain Go1 in response to nitrogen availability. AB - PII-like signal transduction proteins are found in all three domains of life and have been shown to play key roles in the control of bacterial nitrogen assimilation. This communication reports the first target protein of an archaeal PII-like protein, representing a novel PII receptor. The GlnK(1) protein of the methanogenic archaeon Methanosarcina mazei strain Go1 interacts and forms stable complexes with glutamine synthetase (GlnA(1)). Complex formation with GlnK(1) in the absence of metabolites inhibits the activity of GlnA(1). On the other hand, the activity of this enzyme is directly stimulated by the effector molecule 2 oxoglutarate. Moreover, 2-oxoglutarate antagonized the inhibitory effects of GlnK(1) on GlnA(1) activity but did not prevent GlnK(1)/GlnA(1) complex formation. On the basis of these findings, we hypothesize that besides the dominant effector molecule 2-oxoglutarate, the nitrogen sensor GlnK(1) allows finetuning control of the glutamine synthetase activity under changing nitrogen availabilities and propose the following model. (i) Under nitrogen limitation, increasing concentrations of 2-oxoglutarate stimulate maximal GlnA(1) activity and transform GlnA(1) into an activated conformation, which prevents inhibition by GlnK(1). (ii) Upon a shift to nitrogen sufficiency after a period of nitrogen limitation, GlnA(1) activity is reduced by decreasing internal 2-oxoglutarate concentrations through diminished direct activation and by GlnK(1) inhibition. PMID- 15752205 TI - A pathway-specific transcriptional regulatory gene for nikkomycin biosynthesis in Streptomyces ansochromogenes that also influences colony development. AB - DNA sequence analysis of a 7.5 kb XhoI DNA fragment from the region flanking the nikkomycin biosynthesis gene cluster in Streptomyces ansochromogenes revealed one 3.3 kb open reading frame (ORF), designated sanG. The deduced product of sanG (1061 amino acids), which is similar to PimR of Streptomyces natalensis, contains an OmpR-like DNA binding domain in its N-terminal portion and A- and B-type nucleotide binding motifs in the middle of the protein. Disruption of sanG abolished nikkomycin biosynthesis, reduced sporulation and led to brown pigment accumulation. All aspects of this complex phenotype were complemented by a single copy sanG which was integrated into the chromosome. The introduction of multiple copies of sanG resulted in increased nikkomycin production. S1 mapping results indicated that sanG is transcribed from at least three promoters (P1, P2 and P3), P1 being strongly upregulated when production of nikkomycins starts. Two putative transcription units for nikkomycin biosynthesis, starting from sanN and sanO, are dependent on the expression of sanG, whereas a putative transcription unit starting from sanF was not regulated by sanG. These results suggested that sanG encodes a transcriptional activator important for nikkomycin biosynthesis that, unusually, also has pleiotropic effects on secondary metabolism and development. PMID- 15752206 TI - A new in vivo termination function for DNA polymerase I of Escherichia coli K12. AB - Escherichia coli deleted for the tus gene are viable. Thus there must be at least one other mechanism for terminating chromosome synthesis. The tus deletion strain yielded a small fraction of cells that overproduce DNA, as determined by flow cytometry after run-out chromosome replication in the presence of rifampicin and cephalexin. A plasmid, paraBAD tus+, prevented the excess DNA replication only when arabinose was added to the medium to induce the synthesis of the Tus protein. Transduction studies were done to test whether or not additional chromosomal deletions could enhance the excess chromosome replication in the tus deletion strain. A strain containing a second deletion in metE udp overproduced DNA at a high level during run-out replication. Further studies demonstrated that a spontaneous unknown mutation had occurred during the transduction. This mutation was mapped and sequenced. It is polA(G544D). Transduction of polA(G544D) alone into the tus deletion strain produced the high DNA overproduction phenotype. The polA(G544D) and six other polA alleles were then tested in wild type and in tus deletion backgrounds. The two alleles with low levels of 5'-->3' exonuclease (exo) overproduced DNA while those with either high or normal exo overproduce much less DNA in run-out assays in the wild-type background. In contrast, all seven mutant polA alleles caused the high DNA overproduction phenotype in a tus deletion background. To explain these results we propose a new in vivo function for wild-type DNA polymerase I in chromosome termination at site(s) not yet identified. PMID- 15752207 TI - Induction of Staphylococcus epidermidis biofilm formation via proteolytic processing of the accumulation-associated protein by staphylococcal and host proteases. AB - Because of its biofilm forming potential Staphylococcus epidermidis has evolved as a leading cause of device-related infections. The polysaccharide intercellular adhesin (PIA) is significantly involved in biofilm accumulation. However, infections because of PIA-negative strains are not uncommon, suggesting the existence of PIA-independent biofilm accumulation mechanisms. Here we found that biofilm formation in the clinically significant S. epidermidis 5179 depended on the expression of a truncated 140 kDa isoform of the 220 kDa accumulation associated protein Aap. As expression of the truncated Aap isoform leads to biofilm formation in aap-negative S. epidermidis 1585, this domain mediates intercellular adhesion in a polysaccharide-independent manner. In contrast, expression of full-length Aap did not lead to a biofilm-positive phenotype. Obviously, to gain adhesive function, full-length Aap has to be proteolytically processed through staphylococcal proteases as demonstrated by inhibition of biofilm formation by alpha(2)-macroglobulin. Importantly, also exogenously added granulocyte proteases activated Aap, thereby inducing biofilm formation in S. epidermidis 5179 and four additional, independent clinical S. epidermidis strains. It is therefore reasonable to assume that in vivo effector mechanisms of the innate immunity can directly induce protein-dependent S. epidermidis cell aggregation and biofilm formation, thereby enabling the pathogen to evade clearance by phagocytes. PMID- 15752208 TI - Integration and excision by the large serine recombinase phiRv1 integrase. AB - The Mycobacterium tuberculosis prophage-like element phiRv1 encodes a site specific recombination system utilizing an integrase of the serine recombinase family. Recombination occurs between a putative attP site and the host chromosome, but is unusual in that the attB site lies within a redundant repetitive element (REP13E12) of which there are seven copies in the M. tuberculosis genome; four of these elements contain attB sites suitable for phiRv1 integration in vivo. Although the mechanism of directional control of large serine integrases is poorly understood, a recombination directionality factor (RDF) has been identified that is required for phiRv1 integrase-mediated excisive recombination in vivo. Here we describe defined in vitro recombination reactions for both phiRv1 integrase-mediated integration and excision and show that the phiRv1 RDF is not only required for excision but inhibits integrative recombination; neither reaction requires DNA supercoiling, host factors, or high energy cofactors. Integration, excision and excise-mediated inhibition of integration require simple substrates sites, indicating that the control of directionality does not involve the manipulation of higher-order protein-DNA architectures as described for the tyrosine integrases. PMID- 15752209 TI - The Salmonella genomic island 1 is an integrative mobilizable element. AB - Salmonella genomic island 1 (SGI1) is a genomic island containing an antibiotic resistance gene cluster identified in several Salmonella enterica serovars. The SGI1 antibiotic resistance gene cluster, which is a complex class 1 integron, confers the common multidrug resistance phenotype of epidemic S. enterica Typhimurium DT104. The SGI1 occurrence in S. enterica serovars Typhimurium, Agona, Paratyphi B, Albany, Meleagridis and Newport indicates the horizontal transfer potential of SGI1. Here, we report that SGI1 could be conjugally transferred from S. enterica donor strains to non-SGI1 S. enterica and Escherichia coli recipient strains where it integrated into the recipient chromosome in a site-specific manner. First, an extrachromosomal circular form of SGI1 was identified by PCR which forms through a specific recombination of the left and right ends of the integrated SGI1. Chromosomal excision of SGI1 was found to require SGI1-encoded integrase which presents similarities to the lambdoid integrase family. Second, the conjugal transfer of SGI1 required the presence of a helper plasmid. The conjugative IncC plasmid R55 could thus mobilize in trans SGI1 which was transferred from the donor to the recipient strains. By this way, the conjugal transfer of SGI1 occurred at a frequency of 10(-5)-10(-6) transconjugants per donor. No transconjugants could be obtained for the SGI1 donor lacking the int integrase gene. Third, chromosomal integration of SGI1 occurred via a site-specific recombination between a 18 bp sequence found in the circular form of SGI1 and a similar 18 bp sequence at the 3' end of thdF gene in the S. enterica and E. coli chromosome. SGI1 appeared to be transmissible only in the presence of additional conjugative functions provided in trans. SGI1 can thus be classified within the group of integrative mobilizable elements (IMEs). PMID- 15752210 TI - Targeted deletion of Plasmodium knowlesi Duffy binding protein confirms its role in junction formation during invasion. AB - Red cell invasion by Plasmodium merozoites involves multiple steps such as attachment, apical reorientation, junction formation and entry into a parasitophorous vacuole. These steps are mediated by specific molecular interactions. P. vivax and the simian parasite P. knowlesi require interaction with the Duffy blood group antigen to invade human erythrocytes. P. vivax and P. knowlesi Duffy binding proteins (PvDBP and PkDBP), which bind the Duffy antigen during invasion, share regions of sequence homology and belong to a family of erythrocyte binding proteins (EBPs). By deletion of the gene that encodes PkDBP, we demonstrate that interaction of PkDBP with the Duffy antigen is absolutely necessary for invasion of human erythrocytes by P. knowlesi. Electron microscopy studies reveal that PkDBP knockout parasites are unable to form a junction with human erythrocytes. The interaction of PkDBP with the Duffy antigen is thus necessary for the critical step of junction formation during invasion. These studies provide support for development of intervention strategies that target EBPs to inhibit junction formation and block erythrocyte invasion by malaria parasites. PMID- 15752211 TI - Acid-tolerant Listeria monocytogenes persist in a model food system fermented with nisin-producing bacteria. AB - AIMS: To investigate the induction of the acid tolerance response (ATR) in Listeria monocytogenes and to assess the persistence of the pathogen in broth fermented using a nisin-producing starter culture. METHODS AND RESULTS: Lactic, acetic and hydrochloric acids were used to induce the ATR in L. monocytogenes growing at early exponential phase. Cells were then challenged in medium acidified to pH 3.5 with the same acid. Only lactic acid induced a detectable ATR. ATR+ cells maintained their initial numbers after 1 h exposure while ATR- were reduced by c. 4 log10 CFU. ATR+ or ATR- cells were also inoculated in M17G broth fermented with nisin-producing (nis+) or control (nis-) Lactococcus lactis. When exposed to nisin, the numbers of ATR+ cells were c. 2 log10 CFU higher than non detectable ATR- cells at day 3. In the absence of nisin (nis- culture), L. monocytogenes was recovered from all ATR+ and ATR- samples after 30 days. In contrast, no L. monocytogenes were recovered from any nis+ATR- samples but four of five nis+ATR+ samples were positive for L. monocytogenes after 30 days. CONCLUSIONS: The ATR confers cross-resistance to nisin for at least 30 days in a system fermented by nisin-producing bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The cross-resistance induced by the ATR should be considered for the safety of foods fermented with bacteriocin-producing cultures. PMID- 15752212 TI - Controlling production of brominated cyclic depsipeptides by Pseudoalteromonas maricaloris KMM 636T. AB - AIMS: This study aims at evaluating the impact of the nutrient medium components on the in vitro production of the cytotoxic alterochromides. METHODS AND RESULTS: The employment matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) facilitated the identification of a range of brominated cyclic depsipeptides with molecular masses of 843/845, 857/859 and 922/924/926 Da, and 936/938/940 Da produced by the marine bacterium Pseudoalteromonas maricaloris KMM 636T. The fractions of cytotoxic yellow pigments yielded after methanol extraction of P. maricaloris KMM 636T cells grown on five nutrient media were solely composed of brominated cyclic depsipeptides. Bromo-alterochromides A and B were obtained after cultivation on low nutrient media, while dibrominated derivatives were the principal components of the biosynthesis during cultivation on nutrient rich media. CONCLUSIONS: The quantity of bromo-alterochromides and their dibromo- derivates varied depending on the media composition. SIGNIFICANCE AND IMPACT OF THE STUDY: MALDI-TOF mass spectrometry enables to generate accurate mass analysis for the identification of peptide and its derivates which is important in controlling the production of biologically active compounds in vitro. PMID- 15752213 TI - Dehydroepiandosterone induction of increased resistance to vancomycin in Staphylococcus aureus clinical isolates (MSSA, MRSA). AB - AIMS: To investigate whether dehydroepiandosterone (DHEA), an androgen present throughout life, alters the response of Staphylococcus aureus clinical isolates to vancomycin. METHODS AND RESULTS: DHEA in physiologically relevant concentrations (0.1, 0.5, 1.0 and 5.0 micromol l(-1)) was tested for its effect on methicillin-sensitive S. aureus (MSSA, n = 53) and methicillin-resistant S. aureus (MRSA, n = 73) response to vancomycin using standard protocols. Mutant selection was determined by serial transfer of selected isolates (n = 5). DHEA mediated at least a fourfold increase in vancomycin MIC for 42% of MSSA and 21% of MRSA. For five of the isolates (0.1 and 0.5 micromol l(-1) DHEA) the MIC was increased to levels (8 microg ml(-1)) defined as vancomycin-intermediate resistance. CONCLUSION: Resistance was detected only in the presence of DHEA, and was not related to altered generation time, indicating induction of phenotypic resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings require further investigation to determine what role DHEA plays in clinical vancomycin treatment failure that has been reported in the absence of vancomycin genotypic resistance or heteroresistance. PMID- 15752214 TI - Main compounds responsible for off-odour of strawberries infected by Phytophthora cactorum. AB - AIMS: Volatile compounds present in strawberries infected with Phytophthora cactorum, especially those responsible for the characteristic off-odour of such fruits were the subject of this study. METHODS AND RESULTS: Six strawberry varieties (Redgauntlet, Selva, Korona, Tenira, Real, Pegasus) inoculated with P. cactorum strain (PC-5), isolated from naturally infected fruit and one variety inoculated with 15 strains of P. cactorum in the laboratory were analysed. All the samples had a distinct, to a various degree, off-odour reminiscent of watercolour paint with phenolic notes. Volatile compounds were isolated by solid phase microextraction and simultaneous distillation extraction methods. To detect compounds responsible for the characteristic off-odour, gas chromatography olfactometry was used. Two compounds were found to be responsible for the characteristic off-odour of strawberries infected by P. cactorum: 4-ethyl phenol and 4-ethyl-2-metoxy phenol (4-ethyl guaiacol). The content of these compounds in infected varieties ranged from 1.12 to 22.56 mg kg(-1) and 0.14-1.05 mg kg(-1) respectively. Other volatile compounds, not detected in noninoculated sound strawberries, were also identified: camphene, 1-octene-3-ol, 3-octanone, o cymene, phenyl methanol, cis-linaloloxide, nonanal, phenyl ethyl alcohol, 2 undecanone and alpha-muurolene. SIGNIFICANCE AND IMPACT OF THE STUDY: Volatile compounds responsible for the characteristic off-odour of strawberries infected with P. cactorum were identified. Also compounds produced as a result of P. cactorum growth on strawberry fruit were characterized. PMID- 15752215 TI - Phylloplane bacteria increase seedling emergence, growth and yield of field-grown groundnut (Arachis hypogaea L.). AB - AIM: To isolate and characterize groundnut-associated bacterial isolates for growth promotion of groundnut in field. METHODS AND RESULTS: Three hundred and ninety-three groundnut-associated bacteria, representing the geocarposphere, phylloplane and rhizosphere, and endophytes were applied as seed treatment in greenhouse. Maximum increase in plant biomass (up to 26%) was observed following treatment with a rhizosphere isolate identified as Bacillus firmis GRS 123, and two phylloplane isolates Bacillus megaterium GPS 55 and Pseudomonas aeruginosa GPS 21. There was no correlation between the production of L-tryptophan-derived auxins and growth promotion by the test isolates. Actively growing cells and peat formulations of GRS 123 and GPS 55, and actively growing cells of GPS 21, significantly increased the plant growth and pod yield (up to 19%) in field. Rifampicin-resistant mutants of GRS 123 and GPS 21 colonized the ecto- and endorhizospheres of groundnut, respectively, up to 100 days after sowing (DAS), whereas GPS 55 was recovered from both the habitats at 100 DAS. CONCLUSION: Seed bacterization with phylloplane isolates promoted groundnut growth indicating the possibility of isolating rhizosphere beneficial bacteria from different habitats. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of phylloplane bacteria as effective plant growth-promoting rhizobacteria (PGPR) broadens the spectrum of PGPR available for field application. PMID- 15752216 TI - Development of an IMS-PCR assay for the detection of Mycobacterium avium ssp. paratuberculosis in water. AB - AIMS: To develop a sensitive detection method for Mycobacterium avium ssp. paratuberculosis (Map) in water by modifying and optimizing an existing immunomagnetic separation polymerase chain reaction (IMS-PCR) technique. METHODS AND RESULTS: Sterile distilled water (50 ml) spiked with 10(6) Map ml(-1) was subjected to either filtration (0.45 microm pore size) followed directly by IS900 PCR (method 1) or centrifugation (2500 g for 20 min) followed by IMS and IS900 PCR (method 2). Method 2 permitted the detection of Map, whereas method 1 did not. Method 2 was then optimized by adding different concentrations of Tween 80 (0.05, 0.1, 0.2, 0.4 and 0.6% v/v) to water samples spiked with Map (10(6)-1 CFU ml(-1)) prior to centrifugation, and assessing the impact of this action on the detection sensitivity of subsequent IMS-PCR. The optimum Tween 80 concentration was found to be 0.4%, which permitted the detection of 10 Map CFU ml(-1) in spiked water samples by IMS-PCR. CONCLUSIONS: This method will be used to determine the incidence of Map in water destined for domestic use in future studies. SIGNIFICANCE AND IMPACT OF THE STUDY: A sensitive method for the detection of Map in water involving addition of 0.4% Tween 80, centrifugation and IMS-PCR was developed. PMID- 15752217 TI - Evaluation of candidate probiotic strains for gilthead sea bream larvae (Sparus aurata) using an in vivo approach. AB - AIMS: The aim of this study was to evaluate the effect of six bacterial strains on gilthead sea bream larvae (Sparus aurata). METHODS AND RESULTS: Six bacterial strains isolated from well-performing live food cultures were identified by sequencing fragments of their 16s rDNA genome to the genus level as Cytophaga sp., Roseobacter sp., Ruergeria sp., Paracoccus sp., Aeromonas sp. and Shewanella sp. Survival rates of gilthead sea bream larvae transferred to seawater added these bacterial strains at concentrations of 6 +/- 0.3 x 10(5) bacteria ml(-1) were similar to those of larvae transferred to sterilized seawater and showed an average of 86% at 9 days after hatching, whereas, survival rates of larvae transferred to filtered seawater were lower (P < 0.05), and showed an average of 39%, 9 days after hatching. CONCLUSION: Several bacterial strains isolated from well-performing live food cultures showed a positive effect for sea bream larvae when compared with filtered seawater. SIGNIFICANCE AND IMPACT OF THE STUDY: The approach used in this study could be applied as an in vivo evaluation method of candidate probiotic strains used in the rearing of marine fish larvae. PMID- 15752218 TI - Beta-D-glucuronidase activity assay to assess viable Escherichia coli abundance in freshwaters. AB - AIMS: The relationships between the beta-D-glucuronidase (GLUase) activity, the abundance of culturable Escherichia coli and the number of viable E. coli were investigated in river and wastewater samples. METHODS AND RESULTS: GLUase activity was measured as the rate of hydrolysis of 4-methylumbelliferyl-beta-D glucuronide. Culturable E. coli were enumerated by the most probale number (MPN) microplate method. Viable E. coli were estimated by fluorescent in situ hybridization (FISH) coupled with a procedure of viability testing (DVC-FISH procedure). Significant correlations were found between the log of GLUase activity and both, the log culturable E. coli and the log of viable E. coli. CONCLUSIONS: GLUase activity per viable E. coli gave a broadly constant value from low to highly contaminated waters while GLUase activity per culturable E. coli strongly increased at low contaminated waters because of an underestimation of the number of active E. coli by the culture-based method. SIGNIFICANCE AND IMPACT OF THE STUDY: GLUase activity is a reliable parameter for the rapid quantification of viable E. coli in waters. PMID- 15752219 TI - Effect of cereal brans on Lentinula edodes growth and enzyme activities during cultivation on forestry waste. AB - AIMS: To develop strategies for increasing the growth of Lentinula edodes in eucalyptus residues. To this end, we have examined the effects of cereal brans additions on production of mycelial biomass and enzymes. METHODS AND RESULTS: Three isolates of the mushroom shiitake, L. edodes (Berk. Pegler), were evaluated for enzyme and ergosterol production on eucalyptus residue supplemented with 5, 10, 15 and 20% (w/w) of soya, wheat or rice brans. Nitrogen imput on eucalyptus residues accelerated mycelial growth by supplying the L. edodes with this limiting nutrient. High levels of enzymes activities were produced in eucalyptus residues supplemented by soya bran. Comparison of cellulose and xylanase production with manganese peroxidase (MnP) at 20% soya bran indicated that hydrolytic enzymes, but oxidative enzymes were reduced. CONCLUSIONS: Mycelial growth measurements revealed that eucalyptus residues supplemented with cereal brans supported fast growth of L. edodes, indicating that mycelium extension is related to the bioavailability of nitrogen. The type and concentration of nutrient supplement has a considerable effect both on substrate colonization and on the type of hydrolytic and oxidative enzymes produced. These characteristics may be useful for mushroom growing. SIGNIFICANCE AND IMPACT OF THE STUDY: Lentinula edodes is commercially important for edible mushroom production and supplements which enhance growth and enzymes production might also be beneficial for mushroom yields. PMID- 15752220 TI - Molecular vs culture methods for the detection of bacterial faecal indicators in groundwater for human use. AB - AIMS: The current standard culture methods are unable to detect nongrowing bacteria and, thus, might not be sufficient for precise monitoring of the microbiological quality of waters. The use of a molecular method such as PCR could be a valid alternative to detect bacterial faecal contamination indicators such as Escherichia coli and Enterococcus faecalis and reveal the presence of culturable and nonculturable bacterial forms. METHODS AND RESULTS: The presence of E. coli and Ent. faecalis cells in 30 groundwater samples was evaluated with the standard culture method and compared with a specific PCR protocol. A substantial percentage (50%) of the samples not containing culturable cells proved positive in the search for Ent. faecalis DNA by PCR. Quantification by competitive PCR (cPCR) of the DNA detected allowed us to calculate the number of nonculturable cells present in water samples: the number varied from 2 to 120 cells ml(-1). Only four samples were positive for E. coli DNA and the corresponding nonculturable cells varied from 24 to 70 ml(-1). CONCLUSIONS: This study demonstrates that the standard culture methods in use are unable to detect a substantial proportion of the bacterial population which is nonculturable but, as previously demonstrated, potentially still viable and able to express those pathogenic factors needed for causing infections in humans. SIGNIFICANCE AND IMPACT OF THE STUDY: To protect human health it is necessary to develop and use methods which detect the nonculturable as well as culturable bacteria present in water. PMID- 15752221 TI - Comparison of environmental profiles for growth and deoxynivalenol production by Fusarium culmorum and F. graminearum on wheat grain. AB - AIMS: Comparisons were made of the effect of water activity (a(w) 0.99-0.85), temperature (15 and 25 degrees C) and time (40 days) on growth/production of the trichothecene mycotoxin deoxynivalenol (DON) by Fusarium culmorum and Fusarium graminearum on wheat grain. METHODS AND RESULTS: Studies examined colonization of layers of wheat grain for 40 days. Fusarium culmorum grew optimally at 0.98 a(w) and minimally at 0.90 a(w) at 15 and 25 degrees C. Colonization by F. graminearum was optimum at 0.99 a(w) at 25 and 0.98 a(w) at 15 degrees C. Overall, temperature, a(w) and their interactions significantly affected growth of both species. Production of DON occurred over a much narrower range (0.995-0.96 a(w)) than that for growth. Optimum DON was produced at 0.97 and 0.99 a(w) at 15 and 25 degrees C, respectively, by F. culmorum, and at 0.99 a(w) and 15 degrees C and 0.98 a(w) at 25 degrees C for F. graminearum. Statistically, one-, two- and three way interactions were significant for DON production by both species. CONCLUSIONS: This suggests that the ecological requirements for growth and mycotoxin production by such species differ considerably. The two-dimensional profiles on grain for DON production by these two species have not been examined in detail before. SIGNIFICANCE AND IMPACT OF THE STUDY: This type of information is essential for developing climate-based risk models for determining the potential for contamination of cereal grain with this trichothecene mycotoxin. It will also be useful information for monitoring critical control points in prevention of such toxins entering the wheat production chain. PMID- 15752222 TI - Detection of viable and dead Listeria monocytogenes on gouda-like cheeses by real time PCR. AB - AIMS: Surface contamination by Listeria monocytogenes of gouda-like cheeses during processing represents a potential public health problem. The aim of this work was to develop novel real-time PCR diagnostics to detect the presence of viable, dead or viable but not culturable (VBNC) cells on gouda-like cheeses. METHODS AND RESULTS: We used ethidium monoazide bromide (EMA)-PCR for direct quantification of viable and dead cells, while semiquantitative detection of culturable cells below the PCR detection limit (c. 100 CFU g(-1)) was obtained by combining growth and real-time PCR. We were able to quantify the fraction of >0.5% viable cells in a background of dead cells by EMA-PCR, given that the viable cell concentration was above the PCR detection limit. The combined growth and real-time PCR complemented the EMA-PCR, and enabled semiquantitative detection of low levels of culturable cells (10 and 100 CFU g(-1)). SIGNIFICANCE AND IMPACT OF THE STUDY: The significance of this work is that we have developed a novel concept for detection of viable and potentially infectious L. monocytogenes. PMID- 15752223 TI - A time to act for prevention. PMID- 15752224 TI - What have we learned from clinical trials on prevention? AB - The incidence of end-stage renal disease is increasing at an alarming rate. While diabetes is now the most common cause of renal failure, hypertension commonly coexists, exacerbating renal damage and the other vascular complications of diabetes. Clinical trials have shown the benefits of intervention strategies that target not only the retardation of renal disease, but also the primary prevention of the conditions known to cause renal damage. These strategies include: lifestyle modification programs to prevent diabetes and hypertension; aggressive treatment of established hypertension; combination antihypertensive regimens; and the use of angiotensin-converting-enzyme inhibitors and angiotensin II receptor blockers for additional renoprotection beyond that of blood pressure control. In addition, albuminuria and proteinuria have emerged as powerful predictors of progression of renal disease. The present paper reviews the results of available clinical trials and meta-analyses on these interventional strategies, and highlights the potential of albuminuria and proteinuria in predicting renal and cardiovascular outcomes. PMID- 15752225 TI - Nephrotoxicity of several newer agents. PMID- 15752226 TI - Mechanisms of tubulointerstitial injury in IgA nephropathy. AB - BACKGROUND: IgA nephropathy (IgAN) runs a highly variable clinical course, with frequent involvement of tubulointerstitial damage. A subgroup of IgAN with severe tubulointerstitial damage is often associated with the most rapid progression to end-stage renal failure. In IgAN, mesangial sclerosis and tubulointerstitial damage were found to be correlated with the increase in pore size of the glomerular barrier. METHODS: The direct toxicity of proximal tubular epithelial cells (PTEC) by IgA in IgAN is still unresolved. Activation of PTEC by mediators released from infiltrating cells or resident kidney cells that induce tubular inflammation is the common final pathway in most chronic renal diseases. We hypothesize that mediators released from human mesangial cells (HMC) triggered by IgA deposition may lead to PTEC activation. RESULTS: We found that IgA binding to PTEC was less than one tenth that of HMC. The binding was nonspecific and exhibited no increased cell proliferation or enhanced synthesis of cytokines or adhesion molecules. However, when PTEC were cultured with IgA-HMC spent medium prepared from IgAN patients, there was enhanced proliferation of PTEC and increased synthesis of cytokines and adhesion molecules. CONCLUSION: These findings implicate a glomerulotubular cross-talk with mediators released from the mesangium, contributing to the pathogenesis of tubulointerstitial damage in IgAN. There are preliminary data to suggest that the expression of angiotensin II subtype-1 receptor and angiotensin II subtype-2 receptor in PTEC differs from that of HMC. These novel findings may provide clinicians new therapeutic approach for selective blockade of the tubulointerstitial injury in IgAN. PMID- 15752227 TI - Preventing renal failure in patients with severe lupus nephritis. AB - BACKGROUND AND METHODS: Advances in immunosuppressive treatment regimens, with increased efficacy, while minimizing the treatment-related toxicities, and better prevention and treatment of complications, have resulted in improved patient and renal survival in subjects with severe proliferative lupus nephritis over the past few decades. This review discusses the issues that are pertinent to the preservation of renal function in these patients. RESULTS AND CONCLUSION: Treatment of severe proliferative lupus nephritis can be divided into an initial phase of induction followed by a prolonged maintenance phase, both of which impact upon the long-term renal and patient survival. The immunosuppressive potency of the treatment required for disease control varies according to the disease activity during the different phases. Despite variations in the choice, duration, and route of administration of antiproliferative agents, data to date suggest that immunosuppressive treatments combining cyclophosphamide or mycophenolate mofetil with corticosteroid appear to have similar efficacy in terms of inducing immunologic remission. In this regard, the immunologic efficacy of treatment is prerequisite to the prevention of irreversible loss of nephrons, but long-term renal outcome is also dependent on factors other than treatment efficacy, such as preexisting renal parenchymal damage and blood pressure control. Prompt diagnosis, early effective therapy, and reducing the risk of relapses are the disease specific measures that are essential to long-term renal preservation and the prevention of renal failure in subjects with severe proliferative lupus nephritis. PMID- 15752228 TI - Calcification and cardiovascular problems in renal failure. AB - Extraosseus calcification has plagued management of renal failure since the beginning of hemodialysis, but the issue has largely been neglected because the impact on survival was thought to be limited. The recent recognition that hyperphosphatemia is a strong predictor of all-cause mortality, and particularly of cardiac mortality, has transformed the situation. Relatively stringent, though difficult to implement, guidelines have been proposed for the management of hyperphosphatemia. Important recent insights document that, for different reasons, both high and low turnover of bone disease increase the risk of vascular calcifications. Vascular calcification impacts cardiac death not only by complicating coronary atherosclerosis, but also by increasing the stiffness of central arteries, impacting on heart function (increased impedance, reduced coronary perfusion). While in the past extraosseous calcification, including vascular calcification, was thought to be a passive process resulting from transgression of a critical Ca x P product, recent studies show that the adverse effect of hyperphosphatemia is also mediated by active processes (e.g., induction of "osteogenic" genetic programs), and is modulated by calcification inhibitors. PMID- 15752229 TI - The need for early nephrology referral. AB - The incidence of end-stage renal disease is increasing. Progression to end stage can be slowed if kidney damage is detected at an early stage. Prognosis and outcomes in patients with chronic kidney disease have been related to the quality of predialysis care and the timing of referral. Many patients with chronic kidney disease are referred to a nephrologist close to commencement of renal replacement therapy. This leads to suboptimal management of complications of chronic renal insufficiency, and increased morbidity and mortality of patients on renal replacement therapy. This article addresses the evidence that examines the view that patients need to be referred early in order to avoid complications of chronic renal insufficiency. Early referral can be achieved through improved communication between primary health care givers and nephrology services. A multidisciplinary approach has a significant impact on outcomes. In the face of rising incidence of chronic kidney disease, early referral of all patients is not possible. Therefore, identification of patients at risk for rapid deterioration of renal function is important in order to rationalize and reduce health care expenditure. PMID- 15752230 TI - Beneficial responses to modified diets in treating patients with chronic kidney disease. AB - BACKGROUND: In rats, protein-restricted diets prolong survival from experimental chronic kidney disease (CKD). Diets for CKD patients that are restricted in protein and modified to limit the intake of phosphates and sodium not only reduce the accumulation of unexcreted metabolic products but also protect against hypertension and proteinuria. Their ability to slow progression of CKD is controversial, and this fact, plus concern about the nutritional adequacy of restricted diets, has limited this treatment strategy for CKD patients. METHODS: Expected responses of CKD patients to dietary manipulation are examined. The influence of this strategy on complications of CKD, including results from the MDRD Study, is analyzed. RESULTS: Dietary compliance with low-protein diets can be monitored from 24-hour urea excretion. Long-term therapy with diets containing 0.6 g protein/kg/day or less protein supplemented with essential amino acids or their ketoanalogues maintain body weight and normal values of serum proteins while reducing the accumulation of acid, phosphates, urea. Low-protein diets suppress proteinuria even when angiotensin-converting enzyme inhibitors are used. Patients in the MDRD Study who complied with the diet had a significant slowing of the rate of loss of glomerular filtration rate (GFR). CONCLUSION: Successful dietary manipulation can ameliorate many complications of CKD. Low-protein diets are nutritionally safe, they reduce the accumulation of metabolic products, and they can suppress progressive loss of kidney function. Dietary manipulation should be an integral part of the therapy for patients with progressive CKD. PMID- 15752231 TI - Kidney prevention recipes for your office practice. PMID- 15752232 TI - The epidemiology of chronic kidney disease. AB - The world's disease profile is changing, and chronic diseases now account for the majority of global morbidity and mortality, rather than infectious diseases. The causes of chronic kidney diseases reflect this change and diabetes, together with hypertension, is now the major cause of end-stage renal failure worldwide, not only within the developed world, but also increasingly within the emerging world. Diabetes is of epidemic proportions, and its prevalence will double in the next 25 years, particularly in the developing countries. This will place an enormous financial burden on countries, including the cost of the management of end-stage renal failure. Thus, it is medically and economically imperative for awareness, detection, and prevention programs to be introduced across the world, particularly in the developing countries. This will require concerted action from global institutions, governments, health service providers, and medical practitioners. PMID- 15752233 TI - Health economics of renal replacement therapy: perspectives from Singapore. AB - The prevalence of end-stage renal disease (ESRD) in Singapore is high and projected to increase sharply due to the aging population and the high prevalence of diabetes. The number of patients treated with dialysis was projected to rise from 2633 in 1999 to nearly 6000 in the year 2010. The cost of dialysis provision was estimated to increase 2.5-fold from US dollar 90 million in 1999 to US dollar 241 million in 2010. To address this, the Singapore Ministry of Health launched three initiatives in the year 2000: First, to reduce the incidence of ESRD through (1) primary prevention of diabetes, (2) community-based screening to facilitate early detection of patients with diabetes, (3) improving glycemic and blood pressure (BP) control of diabetics in the primary care setting, and (4) the establishment of dedicated Renal Retardation clinics to optimize BP control (and glycemic control for diabetics), and reduce the level of proteinuria in patients at high risk of ESRD development. Second, to increase the percentage of ESRD patients treated with continuous ambulatory peritoneal dialysis (CAPD), which is a lower-cost modality compared with hemodialysis; and third, amendments to the existing opt-out legislation for organ procurement for transplantation to increase the supply of kidneys for cadaveric renal transplant. PMID- 15752235 TI - Proteinuria screening for children. AB - BACKGROUND: In Japan, urine screenings are performed annually at school for proteinuria and hematuria, but the effectiveness of this practice has not been clarified. METHODS: Urine screening at school was performed, and we investigated the prevalence of urine abnormalities and incidence and the causes of their diseases. Therefore, we studied effectiveness of the school-screening program. RESULTS: The prevalence of urinary abnormalities was 0.52% among elementary school children and 0.75% among junior high school children. The incidence was 0.24% among elementary school children. The school-screening program is effective in early detection of glomerulonephritis, so the number of new end-stage renal disease (ESRD) patients starting treatment has been changing. DISCUSSION: The school-screening program is effective for early detection of glomerulonephritis. In case of generations who underwent the school-screening program, the age that one develops ESRD has been rising year by year, and the number of new ESRD patients starting treatment before 20 years old is lower in Japan than in America. CONCLUSION: The school-screening program in Japan represents a highly effective mass screening technique. PMID- 15752234 TI - A report with consensus statements of the International Society of Nephrology 2004 Consensus Workshop on Prevention of Progression of Renal Disease, Hong Kong, June 29, 2004. AB - This report summarizes the discussions of the International Society of Nephrology (ISN) 2004 Consensus Workshop on Prevention of Progression of Renal Disease, which was held in Hong Kong on June 29, 2004. Three key areas were discussed during the workshop: (1) screening for chronic kidney disease; (2) evaluation and estimating progression of chronic kidney disease; and (3) measures to prevent the progression of chronic kidney disease. Fifteen consensus statements were made in these three areas, as endorsed by the participants of the workshop. The ISN can make use of and take reference to these statements in formulating its policy for tackling chronic kidney disease, a disease with significant global impact. PMID- 15752236 TI - The validity of screening based on spot morning urine samples to detect subjects with microalbuminuria in the general population. AB - BACKGROUND: No study has yet investigated the validity of prescreening by albumin measurements in a spot morning urine sample to identify in the general population subjects with microalbuminuria. We therefore tested the diagnostic performance of urinary albumin concentration (UAC) and albumin-creatinine ratio (ACR), measured in a spot morning urine sample, in predicting a urinary albumin excretion (UAE) > or =30 mg in subsequent 24-hour urines (microalbuminuria). METHODS: Subjects (2527) participating in the PREVEND study, a representative sample from the general population, collected a spot morning urine sample and, on average, 77 days later, two 24-hour urine collections. RESULTS: The ROC curve of UAC in predicting microalbuminuria has an area-under-the-curve of 0.92 with a discriminator value of 11.2 mg/L. Using this cut-off value for UAC, sensitivity in predicting microalbuminuria is 85.0%, and specificity 85.0%. For ACR these values are, respectively: area-under-the-curve 0.93, discriminator value 9.9 mg/g, sensitivity 87.6%, and specificity 87.5%. Sensitivity for UAC in predicting microalbuminuria does not differ significantly from the sensitivity for ACR, whereas the difference between the specificities of UAC and ACR reaches statistical significance, but is numerically very small. In various subgroups characterized by differences in urinary creatinine excretion, the area-under-the ROC curve, sensitivity, as well as specificity, do not increase relevantly compared to the results in the overall study population. This holds true for ACR as well as UAC. CONCLUSION: The diagnostic performance of measuring UAC in a spot morning urine sample in predicting microalbuminuria in subsequent 24-hour urine collections is satisfactory, and, moreover, comparable to that of measuring ACR. In order to keep the burden and costs involved in population screening for microalbuminuria as low as possible, we therefore propose prescreening by measuring UAC in a spot morning urine sample. Those subjects with a UAC above a certain predefined level (e.g., 11 mg/L) should be asked to collect timed urine samples. PMID- 15752238 TI - Genetics of common progressive renal disease. AB - Familial aggregation of common chronic kidney diseases provides a unique opportunity to investigate the susceptibility genetic and environmental factors. In the past decade, a wealth of new data has become available concerning the genetic susceptibility leading to numerous nephropathies. Knowledge of the genetic components allows better understanding of initiation and progression of these chronic kidney diseases. In addition, one can envision that identification of genetically susceptible individuals might lead to earlier diagnosis and potential reversal of the current epidemic of end-stage renal disease. The goal of the current discussion is to review various issues pertaining to the role of genetic factors in common chronic kidney diseases, as exemplified by two leading causes of end-stage renal diseases worldwide, nephropathy of type 2 diabetes and IgA nephropathy. The genetic and environmental interplay leading to the nephropathies is highlighted. PMID- 15752237 TI - Prevalence of silent kidney disease in Hong Kong: the screening for Hong Kong Asymptomatic Renal Population and Evaluation (SHARE) program. AB - BACKGROUND: End-stage renal disease (ESRD) is epidemic worldwide. In Hong Kong, the annual incidence of ESRD has risen from 100 pmp (per million population) in 1996 to 140 pmp in 2003. SHARE (Screening for Hong Kong Asymptomatic Renal Population and Evaluation program) is a population-based screening program aimed at identifying the prevalence of unrecognized renal disease in asymptomatic individuals, allowing further evaluation and disease-modifying interventions. METHODS: From November to December 2003, SHARE was conducted in several large residential communities in Hong Kong. The screening tool included a questionnaire documenting demographics and history or family history of diabetes mellitus (DM), hypertension (HT), and chronic kidney disease (CKD), together with on-site measurements of blood pressure (BP) and urine dipstick for protein, blood, and glucose. RESULTS: There were a total of 1811 participants. One thousand two hundred and one subjects were entered into the final analysis. Among the 1201 who were apparently "healthy" (asymptomatic and without history of DM, HT, or CKD), the prevalence of positive (> or =1+) urine dipstick for protein, glucose, blood, protein or blood, any urine abnormality, and HT (BP> or =140/90) was 3.2%, 1.7%, 13.8%, 16%, 17.4%, and 8.7%, respectively. Thirty three percent of the age over 60 years old group had either hypertension or urine abnormalities, compared with 24.0% in the 41- to 60-year-old group and 9.7% in the 20- to 40-year-old group. Having a family history of diabetes or hypertension increases the risk of having urine abnormalities, while a family history of hypertension also increases the risk of high blood pressure. CONCLUSION: It is concluded that subclinical abnormalities in urinalysis or BP readings are prevalent across all age groups in the adult population. An effective screening program at the primary care level that identifies these subjects for further evaluation is warranted, and the public in Hong Kong should be educated toward the significance of such findings in order to have regular health check for asymptomatic renal diseases. PMID- 15752239 TI - Genetic factors in end-stage renal disease. AB - Despite more aggressive treatment of diabetes, hypertension, and hyperlipidemia, the incidence and prevalence rates of end-stage renal disease (ESRD) continue to increase worldwide. The likelihood of developing chronic kidney disease in an individual is determined by interactions between genes and the environment. Familial clustering of nephropathy has repeatedly been observed in all population groups studied and for multiple etiologies of kidney disease. A three- to nine fold greater risk of ESRD is observed in individuals with a family history of ESRD. Marked racial variation in the familial aggregation of kidney disease exists, with high rates in African American, Native American, and Hispanic American families. Disparate etiologies of nephropathy aggregate within African American families, as well. These data have led several investigators to search for genes linked to diabetic and other forms of nephropathy. Evidence for linkage to kidney disease has been detected and replicated at several loci on chromosomes 3q (types 1 and 2 diabetic nephropathy), 10q (diabetic and nondiabetic kidney disease), and 18q (type 2 diabetic nephropathy). Multicenter consortia are currently recruiting large numbers of multiplex diabetic families with index cases having nephropathy for linkage and association analyses. In addition, large scale screening studies are underway, with the goals of better defining the overall prevalence of chronic kidney disease, as well as educating the population about risk factors for nephropathy, including family history. Given the overwhelming burden of kidney disease worldwide, it is imperative that we develop a clearer understanding of the pathogenesis of nephropathy so that individuals at risk can be identified and treated at earlier, potentially reversible, stages of their illness. PMID- 15752240 TI - Diabetic nephropathy: recent insights into the pathophysiology and the progression of diabetic nephropathy. AB - Diabetes has become the single most frequent comorbid condition in patients admitted for renal replacement therapy. This is the result of a greater prevalence of type 2 diabetes and better survival of diabetic patients. Progress has been made in pinpointing the predisposition to diabetes on metabolic abnormalities of muscle mitochondrial metabolism, but the long sought genes predisposing to diabetes and to diabetic nephropathy have not yet been identified. Of great concern are experimental studies documenting that maternal hyperglycemia causes nephron underdosing in the offspring. Relevant to pathogenesis and treatment of diabetic nephropathy are, among others, recent insights that hyperglycemia sensitizes target organs to blood pressure-induced damage, and that local renin-angiotensin systems play an important role in genesis and progression of diabetic nephropathy. PMID- 15752241 TI - Renoprotection with and without blood pressure reduction. AB - BACKGROUND: AT1-receptor blockade dose dependently lowers blood pressure (BP) and albuminuria. Reduction of BP and albuminuria are independent treatment targets for renoprotection, but whether this requires similar dose titration is unknown. METHODS: We tested this in two studies designed to find the optimal antialbuminuric dose of losartan in type 1 diabetic (DM, N= 50) and nondiabetic renal patients (ND, N= 12). After baseline, treatment followed with losartan 50, 100, and 150 mg/day, each dose for eight (DM) or six weeks (ND). At the end of each period, albuminuria (24-hour samples) and mean arterial pressure (MAP) were measured. Patients were divided into "good" and "poor" BP responders (BP+, BP-) according to BP response above or below group median. RESULTS: Baseline MAP in the BP- groups was 102 (97, 104) mm Hg in DM (median, 95% CI) and 91 (80, 108) mm Hg in ND. The top of the dose response for BP (obtained at losartan 100 mg) in the BP- groups was -2 (-4, 3) mm Hg in DM and -1 (-6, 2) mm Hg in ND, versus -15 (-18, -12) mm Hg and -16 (-26, -18) mm Hg in BP+ groups (both P < 0.05). Albuminuria was reduced dose dependently both in BP- and BP+: with 100 mg, the reduction in albuminuria in DM BP- was -32% (-49, 13) versus -45% (-60, -38) in DM BP+ and -45% (-70,-7) versus -25% (-58, -6) in ND BP- and BP+ (all P > 0.05). Moreover, in patients in whom BP fell below the recommended treatment target of 130/80 mm Hg (13 in DM and 10 in ND), albuminuria was progressively reduced, with further increasing the dose of losartan in most patients. CONCLUSION: Absence of BP response to losartan does not preclude a reduction in albuminuria, and optimal reduction of albuminuria may require titration beyond the predefined BP target. PMID- 15752243 TI - Prevention of the progression of chronic kidney disease: practice in China. AB - With the epidemic rise of end-stage renal disease (ESRD) in many countries of the world, there is an urgent need to develop and implement strategies aiming at preventing the development and progression of chronic kidney disease (CKD), and the situation is the same in China. Glomerulonephritis is still the most common cause of ESRD in China; however, epidemiologic studies have revealed that the prevalence of diabetes and hypertension, which both are major causes of ESRD in many developed countries, are increasing dramatically. Additional studies about the prevalence of albuminuria in diabetes mellitus (DM) patients, and the prevalence of kidney lesion in certain high-risk population (e.g., hypertension and atherosclerosis) are undergoing. According to a questionnaire survey and some reports, education program for Chinese nephrologists and practitioners should to be strengthened. PMID- 15752242 TI - Role of lipid control in diabetic nephropathy. AB - Patients with diabetic nephropathy are known to be associated with many lipoprotein abnormalities, including higher plasma levels of very low-density lipoprotein, low-density lipoprotein and triglycerides, and lower levels of high density lipoprotein. Many studies have reported that lipids may induce both glomerular and tubulointerstitial injury through mediators such as cytokines, reactive oxygen species, chemokines, and through hemodynamic changes. Clinical studies in patients with diabetic nephropathy showed that lipid control can be associated with an additional effect of reduction in proteinuria. Experimental studies demonstrated that lipid-lowering agents exerted a certain degree of renoprotection, through both indirect effects from lipid lowering and a direct effect on cell protection. Therefore, lipid control appears to be important in the prevention and treatment of diabetic nephropathy. Diabetic nephropathy has become the leading cause of end-stage renal failure in many countries, including Taiwan. One of the major risk factors for the development and progression of diabetic nephropathy is dyslipidemia. In this paper we will review the role of lipid in mediating renal injury and the beneficial effects of lipid control in diabetic nephropathy. PMID- 15752244 TI - Experience with prevention programs in Thailand. AB - BACKGROUND: Thailand is a developing country with a population of 62 million. It is estimated to have 1.6 million stage 3 or higher chronic kidney disease (CKD) patients. With limited funding for dialysis support, prevention of renal disease progression is mandatory and urgently needed. METHODS: The Nephrology Society of Thailand has conducted a survey among Thai nephrologists. The data were analyzed, and a multidisciplinary team approach was introduced to make patients more informed about CKD and more compliant with dietary prescription. RESULTS: The survey showed that the majority of Thai nephrologists know about CKD guidelines. They regularly employed several modes of drug therapy and appropriate target of blood pressure control. However, only 38% and 5% of nephrologists used multidisciplinary team approach and knew how to calculate dietary protein intake, respectively, in CKD patients. To facilitate dietary counseling and care team approach, we invented several dietary record forms (in Thai), Thai food exchange list, 7-day low-protein Thai food menu, and several patient education guidebooks. We also set 1-day lecture tour courses for physicians, nurses, and dietitians in various parts of the country. At our own institute, we followed 17 CKD patients for 4 years: the creatinine clearance (CCr) and glomerular filtration rate (GFR) were 37.5 +/- 5.5 and 24.4 +/- 4.3 mL/min, respectively, at the beginning, and were 35.7 +/- 4.5 and 25.0 +/- 3.5 mL/min, respectively, 4 years thereafter. The rates of Ccr and GFR decline were -0.5 +/- 0.9 mL/min/year and 0.2 +/- 0.6 mL/min/year, respectively. CONCLUSION: A combination of appropriate drug and dietary therapy by employing a multidisciplinary team approach can effectively delay renal disease progression. PMID- 15752245 TI - Prevention of renal failure: the Malaysian experience. AB - Renal replacement therapy in Malaysia has shown exponential growth since 1990. The dialysis acceptance rate for 2003 was 80 per million population, prevalence 391 per million population. There are now more than 10,000 patients on dialysis. This growth is proportional to the growth in gross domestic product (GDP). Improvement in nephrology and urology services with widespread availability of ultrasonography and renal pathology has improved care of renal patients. Proper management of renal stone disease, lupus nephritis, and acute renal failure has decreased these as causes of end-stage renal disease (ESRD) in younger age groups. Older patients are being accepted for dialysis, and 51% of new patients on dialysis were diabetic in 2003. The prevalence of diabetes is rising in the country (presently 7%); glycemic control of such patients is suboptimal. Thirty three percent of adult Malaysians are hypertensive and blood pressure control is poor (6%). There is a national coordinating committee to oversee the control of diabetes and hypertension in the country. Primary care clinics have been provided with kits to detect microalbuminuria, and ACE inhibitors for the treatment of hypertension and diabetic nephropathy. Prevention of renal failure workshops targeted at primary care doctors have been launched, opportunistic screening at health clinics is being carried out, and public education targeting high-risk groups is ongoing. The challenge in Malaysia is to stem the rising tide of diabetic ESRD. PMID- 15752246 TI - Experience with a program for prevention of chronic renal failure in India. AB - BACKGROUND: For the past eight years, the Kidney Help Trust of Chennai has run a program to prevent chronic renal failure by regular screening of an entire population of 25,000, and treatment of diabetes and hypertension with the cheapest available drugs. The total cost amounts to 25 cents per capita of the study population. METHODS: The program has recently been expanded to cover the adjacent area with a population of 21,500. Both the original population and the new population are being surveyed. Persons at risk of renal failure are identified as reported earlier, and glomerular filtration rate (GFR) is estimated by the MDRD formula. The survey is as yet incomplete. Six thousand one hundred people in the new area, and 20,986 in the old, have been studied so far, and the numbers and percentage of those with GFR below normal have been compared. RESULTS: The prevalence of impaired renal function (GFR below 80 mL/min) in the original population is 8.6 per thousand, and in the new population is 13.9 per thousand (P= 0.005, RR 1.61, CI 1.15-2.24). CONCLUSION: This model provides an effective method for prevention of chronic renal failure at the community level. PMID- 15752247 TI - Pathophysiology of disease progression in proteinuric nephropathies. PMID- 15752248 TI - The scientific care for prevention: an overview. PMID- 15752249 TI - Blockade of NFkappaB activation and renal inflammation by ultrasound-mediated gene transfer of Smad7 in rat remnant kidney. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) in renal fibrosis has been well studied, but little attention has been paid to the potential role of TGF beta in the resolution of renal inflammation. We hypothesize that TGF-beta exerts its anti-inflammation properties by stimulating its negative signaling pathway involving Smad7. METHODS: A rat remnant kidney model was treated with a doxycycline-regulated Smad7 gene or control empty vector using an ultrasound microbubble (Optison)-mediated system. Smad7 transgene expression within the kidney was tightly controlled by the addition of doxycycline in the daily drinking water. All animals were euthanized at week 4 for examination of inflammatory responses. RESULTS: Real-time polymerase chain reaction (PCR) and immunohistochemistry revealed that gene transfer of Smad7 resulted in a substantial inhibition of interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) expression (all P < 0.01 vs. control). This was associated with the attenuation of histology damage, proteinuria, serum creatinine, and an increase in creatinine clearance (all P < 0.05). In addition, overexpression of Smad7 significantly inhibited renal inflammation, including ICAM-1, iNOS, and accumulation of macrophages and T cells in both glomeruli and tubulointerstitium. Furthermore, gene transfer of Smad7 also substantially blocked nuclear factor kappa B (NFkappaB) activation in the rat remnant kidney (P < 0.01). CONCLUSION: TGF-beta/Smad7 signaling plays a critical role in the resolution of renal inflammation in rat remnant kidney model. Inhibition of NFkappaB activation is a key mechanism by which Smad7 suppresses renal inflammation, which suggests a crosstalk pathway between NFkappaB and Smad7. The ability of Smad7 to inhibit renal inflammation indicates that ultrasound-microbubble-mediated Smad7 gene therapy may represents a new therapeutic strategy for glomerulonephritis. PMID- 15752250 TI - Basic research in progressive glomerulopathies: the role of fibrosing factors in IgA nephropathy and diabetic nephropathy. PMID- 15752251 TI - The role of tubulointerstitial inflammation. AB - BACKGROUND: Exploration of the role of tubulointerstitial inflammation in experimental chronic renal disease (CRD) is an essential step to understanding and finding new treatments for human CRD. Adriamycin nephrosis (AN) is an experimental analogue of human focal glomerular sclerosis and tubulointerstitial inflammation. METHODS: Using murine and rat AN, we have systematically investigated the pathogenic roles of chemokines, costimulatory molecules, and inflammatory cells, such as macrophages and effector and regulatory T lymphocytes. The profile of humoral and cellular mediators was studied in vitro and in vivo. The pathogenic significance of various factors was investigated by DNA vaccination, leukocyte reconstitution and depletion, retroviral transduction, and blockade with monoclonal antibodies. RESULTS: Renal cortical and tubular cell CC-chemokines, including MCP-1, RANTES, and MIP-1alpha, were up-regulated via mediation of NFkappaB, and contributed to disease by attracting inflammatory cells into the interstitium. The role of these chemokines was confirmed by DNA vaccination. CD40-CD40L costimulation signals were involved in expansion and activation of the inflammatory infiltrate, whereas PD-1 signals were inhibitory, and CD28-B7 appeared to have a neutral effect. Macrophage and CD8+ T cells were shown to be effectors of injury, whereas CD4+CD25+ and gammadelta T cells acted as regulatory cells. FoxP3 transduction was able to convert naive T cells to CD4+CD25+ regulatory T cells. CONCLUSION: There is a broad range of humoral and cellular factors involved in the pathogenesis of experimental CRD, some of which are potential targets for treatment of human CRD. PMID- 15752252 TI - Randomized-controlled trial: effect of a reservoir biteguard on quality of life in xerostomia. AB - BACKGROUND: To assess the effect of a reservoir biteguard for artificial saliva on the oral health-related quality of life of patients with xerostomia. METHODS: Double-blind randomized placebo-controlled trial among 86 adults with xerostomia. Study group received the trial biteguard. Control group received a conventional biteguard. Outcomes were number of impacts and total scores as recorded by oral impacts on daily performances (OIDP). RESULTS: At 1-month follow up 84 people remained in the trial. The median number of impacts in the study and control groups was 3 and 4 respectively. The median total score was 6 and 12 respectively. In ANCOVA receipt of the reservoir biteguard reduced the number of impacts recorded by OIDP but there was no difference in the total score. CONCLUSIONS: Reservoir biteguards improved the quality of life of people with xerostomia by reducing the number of impacts on daily life. PMID- 15752253 TI - Assessment of SS-A and SS-B in parotid saliva of patients with Sjogren's syndrome. AB - BACKGROUND: The purpose of this study was to compare the sensitivity of parotid saliva to that of serum in detecting anti-SSA/Ro and anti-SSB/La autoantibodies in patients with Sjogren's syndrome. METHODS: Forty patients and 20 controls participated in the study; all patients met the 1993 European Community criteria for the diagnosis of Sjogren's syndrome. Healthy controls were age- and sex matched individuals with no signs or symptoms of Sjogren's syndrome. Serum and saliva samples were evaluated using AffiniTech SSA/Ro and SSB/La antibodies kits (AffiniTech, Ltd. Bentonville, AR, USA). The results were also compared with serological status of SS-A and SS-B as reported by an independent clinical laboratory. RESULTS: Serum was significantly more sensitive than saliva in detecting SSA/Ro and SSB/La antibodies (P = 0.001). There was high agreement between the results with the AffiniTech kits and the independent laboratory (kappa = 0.80; P < 0.001). However, there was poor agreement between saliva and serum results (kappa = 0.174; P = 0.168). CONCLUSIONS: The overall results appear to support that serum analysis is effective method for evaluating the presence of SS-A and SS-B autoantibodies. PMID- 15752254 TI - Hepatitis C virus-associated oral lichen planus: is the geographical heterogeneity related to HLA-DR6? AB - BACKGROUND: The association between hepatitis C virus (HCV) and oral lichen planus (OLP) is more common in the Mediterranean area and Japan, possibly because of immunogenetic factors. METHODS: Intermediate-resolution HLA-DRB typing by hybridization with oligonucleotide probes was performed in 31 Italian OLP patients with HCV infection, in 45 Italian OLP and in 48 British OLP patients without HCV infection. As healthy controls we included data from 145 unrelated Italian and 101 unrelated British bone marrow donors. RESULTS: Italian HCV+ve OLP patients possessed the HLA-DR6 allele more frequently than Italian and British OLP patients without HCV infection (51.6% vs. 17.7% vs. 16.7%; P corrected = 0.028 and 0.017, respectively). There was no difference in the frequency of the HLA-DR6 allele between Italian and British control subjects. CONCLUSIONS: The present data suggest that HLA-DR6 may be responsible for the peculiar geographic heterogeneity of the association between HCV and OLP. PMID- 15752255 TI - Frequent microsatellite alterations of chromosome locus 4q13.1 in oral squamous cell carcinomas. AB - BACKGROUND: Studies have revealed that losses of chromosome 4q24-25 regions are frequent in cancers including head and neck squamous cell carcinoma. Our previous comparative genomic hybridization analysis showed extensive losses of chromosome arm 4q in oral squamous cell carcinoma (OSCC). METHODS: To be more precise in mapping the potential regions of allelic losses and to understand the microsatellite instability (MSI) on 4q involving in oral pathogenesis, we performed allelotypings using eight polymorphic markers. Microsatellite analyses were first performed on 100 randomly selected controls to confirm the high informative rates of markers. Twenty OSCC tissues were microdissected from surgical specimens for microsatellite alterations (MA) analysis. RESULTS: MA was observed in 95% OSCC cases. The most eminently altered locus was 4q13.1 (75%), followed by 4q22.2 and 4q32.1 (55%). Allelic losses also occurred most frequently on these loci. Thirty-five percent cases had MA spanning 4q13.1 to 4q21.1. MSI occurred in 35% OSCC, at a lesser extent compared with allelic losses. The most common locus for MSI was 4q21.2 (20%). In addition, 4q MSI was significantly associated with the lymph node metastasis of OSCC (P = 0.01). So far, most tumor suppressor genes on 4q have not been specified. CONCLUSION: Our results were additive to previous findings and proposed novel scenario of suppressor loci located at 4q13.1-21.1 whose inactivation could be important for progression of OSCC. PMID- 15752256 TI - Morphological changes of regional lymph node in squamous cell carcinoma of the oral cavity. AB - BACKGROUND: Regional immune responses with various types of cancer have been studied histopathologically, however, the prognostic value remains conflicting. The aim of this study was to evaluate morphological changes related to lymph node metastasis and the prognostic value for oral cavity squamous cell carcinoma. METHODS: With histopathologic whole architecture of 430 lymph nodes, gross area, germinal center (GC) area, paracortical area (PA), and tumor area were measured. RESULTS: Metastatic node had significantly lower distribution ratio of PA to lymphoarea than that of tumor-free node. GC area was not constantly associated with lymph node metastasis. In Cox multivariate analysis, the mean ratio of PA to gross area/lymphoarea was an independent prognostic factor. CONCLUSIONS: The proportion of PA to gross/lymph area was associated with lymph node metastasis and long-term survival and may be useful in stratification of those patients for a requirement of adjuvant treatments. PMID- 15752257 TI - Expression of p63 and p73 in ameloblastomas. AB - BACKGROUND: To clarify the role of p53 homologs in oncogenesis and cytodifferentiation of odontogenic tumors, expression of p63 and p73 was analyzed in ameloblastomas as well as tooth germs. METHODS: Tissue specimens of nine tooth germs and 48 benign and five malignant ameloblastomas were examined by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) for the expression of p63 and p73. RESULTS: Immunoreactivity for p63 and p73 was evident in epithelial cells neighboring the basement membrane in developing and neoplastic odontogenic tissues. p63 expression in desmoplastic ameloblastomas was significantly higher than in acanthomatous and granular cell ameloblastomas, and ameloblastic carcinomas showed higher p63 expression than metastasizing ameloblastomas. p73 expression was significantly higher in plexiform ameloblastomas than in follicular ameloblastomas, and basal cell ameloblastomas showed higher p73 expression than granular cell ameloblastomas. mRNA transcripts for Delta Np63 and TAp73 were detected in all developing and neoplastic odontogenic tissues. TAp63 mRNA was expressed in five of eight tooth germs, 16 of 34 ameloblastomas, and one of one malignant ameloblastoma, whereas Delta Np73 mRNA was recognized in one of eight tooth germs, nine of 34 ameloblastomas, and one of one malignant ameloblastoma. CONCLUSION: The expression of p63 and p73 suggests that these p53 homologs play a role in differentiation and proliferation of odontogenic epithelial cells. Variations of predominantly expressed isoforms suggest that p63 and p73 might differentially function in odontogenic tissues. PMID- 15752258 TI - Ploidy analysis of field cancerization and cancer development in the hamster cheek pouch carcinogenesis model. AB - BACKGROUND: The hamster cheek-pouch carcinogenesis model is a well-known animal system that closely mimics the development of premalignant and malignant lesions in human oral cancer. Our aim was to numerically characterize the premalignant and malignant lesions and expressions of field cancerization in this model using ploidy as the end-point. METHODS: To study the DNA content and proliferation status of the cells in this model we assessed the Feulgen reaction and the immunohistochemical reaction for 5-bromo-2-deoxiuridine (BrdU) in different histological areas of serial tissue sections of the cheek pouches of animals injected with BrdU. RESULTS: Ploidy values were higher in cancerized epithelia with no unusual microscopic features (NUMF), in preneoplastic and tumor areas than in control epithelia. The aneuploidy index was higher in NUMF areas than in control and differed significantly from control in preneoplastic areas and carcinoma. CONCLUSIONS: The unexpected alteration in DNA content observed in NUMF epithelia is of great relevance as a biomarker of field cancerized areas. PMID- 15752259 TI - Expression of p63 protein and mRNA in oral epithelial dysplasia. AB - BACKGROUND: Abnormalities in the TP53 are regarded as the most consistent findings in oral squamous cell carcinoma. Two related members of the TP53 family, p73 and p63, have shown remarkable structural similarity to TP53, indicating possible functional and biological interactions. The aim of the present study was to investigate the expression of p63 protein and mRNA in oral epithelial dysplasia. METHODS: Immunohistochemical p63 staining was compared for samples from 90 male patients with buccal epithelial dysplasias and 15 healthy individuals with normal buccal mucosa and 15 subjects with reactive epithelial hyperplasia of the oral mucosa secondary to traumatic insult. The buccal lesions consisted of mild, moderate and severe epithelial dysplasias (30 samples in each category). The mRNA expression using reverse transcription polymerase chain reaction (RT-PCR) was also included for a subset of available fresh tissue specimens (four samples in each category of mild and moderate epithelial dysplasia; five samples in severe epithelial dysplasia; five samples in each of normal and reactive epithelial hyperplasia). RESULTS: Nuclear p63 staining was demonstrated predominantly in the basal layers of the epithelium of the normal buccal mucosa and reactive epithelial hyperplasia specimens. For epithelial dysplasia lesions, however, staining was not restricted to the basal layers, extending to the middle spinous layer for samples in the mild category, with p63 immunoexpression observed across almost the full thickness of the dysplastic epithelium for analogous moderate and severe specimens. Compared with normal/reactive hyperplastic mucosa, p63 staining in the dysplastic mucosa was significantly increased. The severity of dysplasia was increased with the increase of p63 staining. Furthermore, Delta Np63mRNA was identified in all of the fresh tissue samples whereas expression of transactivation (TA) isotype was not detected. A subset of moderate epithelial dysplasia and severe variant showing p63-positive staining has undergone malignant transformation to squamous cell carcinomas in about 5 years follow-up. CONCLUSION: Our results indicate that impaired p63 immunoexpression (predominantly Delta N isoform) is associated with the severity of oral epithelial dysplasias and up-regulation of p63 may play a role in the early stage of human oral tumorigenesis. PMID- 15752260 TI - An ultrastructural and a cytochemical study of candidal invasion of reconstituted human oral epithelium. AB - BACKGROUND: Opportunistic yeast, Candida albicans causes superficial and systemic mycoses in compromised patients. Adhesion to host tissues, morphogenesis and extracellular phospholipases (PL) are thought to contribute to its virulence. The nature of numerous host-parasite interactions at the invasive phase of oral candidiasis is not fully understood. Hence in this study, we explore the ultrastructural features of oral candidiasis using a tissue culture model based on reconstituted human oral epithelium (RHOE). METHODS: Reconstituted human oral epithelium (Skinethic Laboratory, Nice, France) was inoculated with C. albicans SC5314 and incubated up to 48 h. The infected tissue was harvested at 12, 24 and 48 h and examined using light, scanning (SEM) and transmission electron microscopy (TEM). Localized activity of PLs of C. albicans during tissue invasion was also examined using a cytochemical method. RESULTS: Over a period of 48 h C. albicans invaded the RHOE, and histological examination revealed characteristic hallmarks of pathological tissue invasion. Hyphal penetration into the superficial epithelium, particularly at cell junctions, together with features of cellular internalization of yeasts was noted. Phospholipase activity was visible at the tips of hyphae and initial sites of bud formation. Further, SEM studies revealed cavitations on the surface epithelial cells particularly pronounced at the sites of hyphal invasion. Hyphal invasion was seen both at cell surfaces and intercellular cell junctions of the epithelium, the latter resembling thigmotropic behaviour. CONCLUSIONS: Our findings confirm that multiple cellular interactions such as internalization, thigmotropism and extracellular PLs contribute to invasive candidiasis. The RHOE model, described here, appears to be a satisfactory model for the investigation of ultrastructural and histochemical features of invasive candidiasis in humans. PMID- 15752261 TI - The phenotype of in vitro reconstituted normal human oral epithelium is essentially determined by culture medium. AB - OBJECTIVE: To evaluate the role of various culture media and serum supplement on growth of oral cells in monolayer, and on morphogenesis of in vitro reconstituted normal human oral epithelium. METHODS: Primary keratinocytes and fibroblasts were isolated from normal human buccal mucosa. The monolayers were assessed by growth curve analysis and morphology. The organotypic cultures were evaluated by morphometry, immunohistochemistry, and TUNEL. RESULTS: FAD medium (a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium) was able to support fibroblast growth in defined conditions, and to diminish the negative effect of physiological Ca concentration on keratinocytes in monolayers. Medium type had a profound influence on morphogenesis of in vitro reconstituted human oral epithelium. FAD medium was superior to other types of medium tested in supporting both epithelial growth and differentiation. Defined conditions supported epithelial morphogenesis equally well as serum-containing medium. CONCLUSIONS: This study points to an essential role of medium composition for optimized growth and differentiation of primary organotypic cultures. PMID- 15752262 TI - Successful treatment of an extensive verrucous carcinoma with topical 5 aminolevulinic acid-mediated photodynamic therapy. AB - Our recent study found that a new topical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) protocol composed of multiple 3-min fractionated irradiations with a light emitting diode (LED) red light at 635 +/- 5 nm for a total of 1000 s (fluence rate: 100 mW/cm(2); light exposure dose: 100 J/cm(2)) after topical application of 20% ALA for 1.5 or 2 h can be used successfully for the treatment of oral verrucous hyperplasia. In this case report, we tested the efficacy of this new treatment protocol of ALA-PDT for an extraoral verrucous carcinoma (VC) lesion at the right mouth angle and an intraoral VC lesion at the right buccal mucosa of a 56-year-old male areca quid chewer and smoker. The extraoral tumor was cleared after six treatments of topical ALA-PDT and the intraoral tumor showed complete regression after 22 treatments of topical ALA PDT. No recurrence of the VC lesion was found after a follow-up period of 6 months. We suggest that PDT using a topical application of 20% ALA followed by multiple 3-min fractionated irradiations with an LED red light is also an effective and successful treatment modality for VC. PMID- 15752263 TI - Strain typing of German transmissible spongiform encephalopathies field cases in small ruminants by biochemical methods. AB - Following the implementation of a large scale transmissible spongiform encephalopathies (TSE) surveillance programme of small ruminants, evidence for a natural transmission of bovine spongiform encephalopathy (BSE) to a French goat has been found. During the years 2002-2004, a massive TSE rapid testing programme on >250,000 small ruminants was carried out in Germany. In this national survey, 186 scrapie-affected sheep were found which originated from 78 flocks. The majority of these cases were of the classical TSE type (115 sheep belonging to 14 outbreaks). However, 71 cases coming from 64 flocks were of the novel atypical scrapie type. According to the regulation EU 999/2001, all TSE cases in small ruminants have to be examined by strain typing methods to explore any possibility of the existence of BSE cases in the field sheep population. Here we report on a biochemical typing strategy (termed FLI-test), which includes the determination of molecular masses, antibody binding affinities and glycosylation pattern of the TSE induced abnormal prion protein. Based on this typing approach none of the analysed German classical TSE outbreaks (total number of analysed sheep: 36) displayed biochemical features indicative for a BSE infection. However, in two cases distinct but BSE-unrelated PrP(Sc) types were found, which alludes to the existence of different scrapie strains in the German sheep population. PMID- 15752264 TI - Detection of beak and feather disease virus (BFDV) and avian polyomavirus (APV) DNA in psittacine birds in Italy. AB - Beak and feather disease (psittacine circovirus) and Budgerigar fledgling disease (avian polyomavirus) are viral diseases that can frequently affect captive psittacine birds. We designed the first survey to investigate the presence of beak and feather disease virus (BFDV) and Avian polyomavirus (APV) inside the population of captive psittacine birds in Italy. Samples were collected in 18 Italian psittacine breeding centres and four trade centres over a 4-year period. A total of 1516 birds were tested for BFDV and 877 birds were tested for APV by means of a polymerase-chain-reaction (PCR) assay. BFDV was found in 122 (8.05%) and APV in 7 (0.79%) birds. No significant difference in infection rate was found between imported and locally raised parrots. We report the first BFDV DNA isolation in wild birds imported to Italy from Papua New Guinea. PMID- 15752265 TI - Effects of fluorequinolone treatment acidified feed, and improved hygiene measures on the occurrence of Salmonella Typhimurium DT104 in an integrated pig breeding herd. AB - Worldwide, the use of antimicrobials in food production has been associated with drug resistance in foodborne pathogens such as Salmonella. However, little is known about the efficaciousness of fluorequinolone treatment on Salmonella Typhimurium T104 infections in pig breeding herds. A combined eradication procedure with enrofloxacin application on sows and piglets, feeding of encapsulated organic acids to sows, disinfection with peracetic acid, separation of the growers from the sows and serological discrimination using a new whole cell-based enzyme-linked immnosorbent assay (ELISA) was evaluated for the suitability to eradicate and to control endemic S. Typhimurium DT104 infections in a closed herd. Thirty-seven sows and their piglets were treated everyday from day 14 ante partum until the day of weaning. Eighteen sows and their piglets served as controls. From the first day of life until day 168 after birth, faecal samples (n = 1671) of all piglets were analysed for Salmonella shedding. In parallel, systemic antibody responses were monitored by whole-cell-based isotype specific ELISA systems. From birth to weaning the prevalence in both groups was between 2% and 9%. After weaning, intermittent shedding could be observed in both groups, and salmonellae could be found in up to 7.7% of the faecal samples. As a result, a dramatic increase in Salmonella-infected growers was observed, as of day 115 after birth, 47.4% of the animals of the treated group were tested positive for S. Typhimurium. Our results indicate that despite long-term antibiosis treatment and optimized hygiene measures, shedding of S. Typhimurium by the sows and the subsequent infection of their offspring could not be effectively prevented. Although it could be not shown that elimination of S. Typhimurium DT104 infection was achieved, the disinfection procedures described and the diagnostic test used are effective instruments to decrease the Salmonella load and to identify individual infected animals. Both of these are important factors for an improved consumer protection. PMID- 15752266 TI - The use of homologous antigen in the serological diagnosis of brucellosis caused by Brucella melitensis. AB - In the European Union the serological diagnosis of brucellosis caused by Brucella melitensis is performed using the heterologous antigen of B. abortus S99. The possible higher sensitivity or ability of an early detection of antibodies by a homologous antigen may prove very useful in the final phases of an eradication programme. Results obtained in sheep experimentally infected by B. melitensis biovar 3 were compared using B. abortus S99, B. melitensis M1, M2 and M3 antigens in the Rose Bengal plate test (RBPT), the complement fixation test (CFT) and an enzyme-linked immunosorbent assay (ELISA) test. Forty-six sheep from an officially brucellosis-free flock were experimentally infected intraconjunctivally with B. melitensis biovar 3. Prior to infection, all animals were tested first against Brucella antibodies, weekly for 2 months post-infection (PI) and then monthly for a further 7 months. All sera were tested against the antigens listed above using RBPT, CFT and ELISA. Using a Bayesian approach, test sensitivities were estimated and compared. Their ability for the early detection of antibodies was evaluated through a regression model based on a logit response model, using the number of days PI as the independent variable and the logit of the fraction of positive animals as the dependent variable. No significant differences were detected among the various antigens used, either in terms of sensitivity or in terms of antibody kinetics; however, the CFT was significantly less sensitive than the RBPT and ELISA and it also showed a lower rate of increase of percentage positive animals (beta-coefficient of regression analysis). PMID- 15752267 TI - Isolation and characterization of potentially human pathogenic, cytotoxin producing aeromonas strains from retailed seafood in Berlin, Germany. AB - The presence of potentially human pathogenic strains of Aeromonas was investigated in 84 samples of seafood which were purchased from retail traders in Berlin, Germany in spring 2000. A total of 134 Aeromonas strains were isolated on selective [GSP agar and Aeromonas (Ryan) agar] and unselective (standard count agar and enterohaemolysin agar) media from 27 (32.1%) of the samples and were classified as Aeromonas hydrophila (67.9%), A. caviae (26.1%) and A. sobria (6.0%) by biotyping. Thirteen (48.1%) of the 27 positive samples contained more than one species of Aeromonas. Production of haemolysins on enterohaemolysin agar was found with 132 (98.5%) of the strains at 28 degrees C and with 130 strains (97.0%) at 37 degrees C growth temperature. Vero cytotoxins were produced by 99 (73.9%) of the strains when grown at 28 degrees C but only by 24 of the strains (17.9%) at 37 degrees C. The latter strains were identified as A. hydrophila (n = 22) and A. sobria (n = 2) which came from 17 (20.2%) samples of raw seafood and from ready-to-eat salted herring 'Matjes' products. Cytotoxin-encoding genes for aerolysin (aer) and haemolysin A (hlyA) were investigated by PCR. Aer and hlyA genes were detected in both, strains which produced toxins only at 28 degrees C and strains which produced toxins at 37 degrees C. Our data indicate that raw seafood and ready-to-eat fish products can harbour potential human pathogenic, cytotoxin producing Aeromonas strains. PMID- 15752268 TI - The use of beef bull semen reduced the risk of abortion in Neospora-seropositive dairy cows. AB - There is an evidence that the epidemiology of neosporosis differs in dairy and beef cattle, such that beef cattle carry a lower risk of abortion. The aim of the present study was to establish whether artificial insemination using semen from beef bulls could reduce the risk of abortion in dairy cows seropositive for the Neospora caninum parasite. Our study was based on yearly serological screening for neosporosis and on the confirmation of Neospora infection in aborted fetuses in two high-producing dairy herds with a mean 28% seroprevalence of N. caninum antibodies. The study population comprised of 273 pregnancies in seropositive animals: 156 pregnancies monitored after insemination using Holstein-Friesian semen and 117 after insemination using beef bull semen. Abortion rates for these animals were 28.2% (77 of 273), 34.6% (54 of 156) and 19.7% (23 of 117). Logistic regression analysis indicated no significant effects of lactation number and previous abortion on the abortion rate. Based on the odds ratio, a 1-unit increase in the Neospora antibody titre yielded a 1.01-fold increase in the abortion rate. The likelihood of abortion was two times higher for cows in one of the two herds and 2.8 times lower (one of 0.36) for pregnant cows inseminated with beef bull semen rather than Holstein-Friesian semen. Our results indicate that the use of beef bull semen can reduce the risk of abortion in dairy cows, and suggest that annual screening for neosporosis, specifically the antibody titre to the protozoan, could be an useful predictor of abortion risk in reproductive health programmes. PMID- 15752269 TI - Occurrence of Isospora suis in Germany, Switzerland and Austria. AB - Nationwide surveys for the occurrence of Isospora suis were carried out in Germany, Austria and Switzerland including a questionnaire regarding herd size, health status and management practices and a coccidiosis sampling kit for pooled faecal samples from litters of suckling piglets. A total of 184 veterinary practices participated in the survey and returned 1745 samples (331 kits) from 324 farms in the north (n = 98), south (n = 84), centre/east (n = 42) and west (n = 10) of Germany, Austria (n = 61) and Switzerland (n = 29) with larger farms in north and centre/east (average number of sows: 270 and 500) and smaller ones in the south (95), Austria (60) and Switzerland (43). Larger farms tended to have better hygienic standards (slatted floors, disinfection of the farrowing units). The majority of the participating farms (93.5%) reported problems with diarrhoea in piglets at 2-3 weeks of age, significantly associated (P < 0.001) with uneven weaning weights (94.9%). Toltrazuril (5%; Baycox) was used only rarely; however, in these farms unevenness of weaning weights was less frequently observed (P = 0.011). A 76.2% of the farms were positive for I. suis (samples contained mostly low or moderate oocyst numbers), especially in the south (P < 0.001). Oocysts were more frequently found in samples from farms with reported diarrhoea (P = 0.011), uneven weight gain (P = 0.019) or in herds of small size (P < 0.001). Disinfection, floor type or treatment with toltrazuril did not affect the frequency of observation of oocysts. PMID- 15752270 TI - The detection of an unidentified type of adenovirus in the stools of calves with weak calf syndrome by use of a commercial kit designed for the detection of human adenoviruses. AB - An outbreak of polyarthritis in newborn calves in a large collective dairy herd was characterized by intra-articular blood-tinged synoviae, blood tainted faeces and massive subcorneal haemorrhages. Faecal samples from eight clinical newborn cases, 10 from unrelated dairy farms and 10 faecal samples from healthy calves were examined by the Rida Quick rotavirus/adenovirus-combi test . A specific adenovirus antigen precipitin-line was seen in the reaction in all the faecal samples from the diseased calves (n = 8), while all the others (n = 20) were negative. In addition, the same positive reaction was noted when one aqueous humor and two synovial samples were tested with this kit. Several other enteropathogens were found sporadically, but no conclusive significance could be attributed to their presence. Bovine viral diarrhoea and infectious bovine rhinothracheitis viruses as well as Chlamydia spp. and Mycoplasma spp. were not involved in this episode. PMID- 15752271 TI - Shortening of the bovine tongue according to regulation (EC) 999/2001 is not complying with the current legal definition of specified risk material - a macroscopical and histological preliminary study. AB - The full elimination of all specified risk material (SRM) in food of animal origin is crucial for consumer protection and is of high priority in inner EU trade. Among other tissues, the tonsils of cattle are considered as SRM. The aim of this study was to evaluate whether the 'cut at the back of the tongue just before the tongue bones' required by EC regulation is sufficient to remove tonsils and lymphatic tissue completely. Eight skulls from cattle were collected for the simulation of a vertical cut according to the EC regulation and the detection of the target at the back of the tongue. Further, specimens of the lingual mucosa were cut out from two tongues and examined microscopically. The most caudal of these specimens was from the macroscopically visible part of the lingual tonsil. The most rostral specimen contained the most caudal Papilla vallata. Simulation of the obligatory ventro-dorsal cut yielded hits at varying locations on the dorsal surface of the tongue, sometimes including tissue of the lingual tonsil. Histological examination of the lingual mucosa gave clear evidence that lymphatic tissue resembling the tissue of a tonsil in terms of its histological organization and infiltration of the mucosal epithelium could even be found in areas with no macroscopically visible lingual tonsils. PMID- 15752272 TI - Ultrastructure of Goussia cruciata (Apicomplexa: Coccidia) infecting the liver of horse mackerel, Trachurus trachurus (L.), from Ibero-Atlantic waters. AB - The ultrastructure of developmental stages of Goussia cruciata and the pathology they cause in the liver of Trachurus trachurus (Teleostei: Carangidae) caught off the Galician (North-West Spain) and Portuguese North Atlantic coasts are described. Each oocyst contained four ellipsoidal sporocysts, with two sporozoites. The sporocyst wall consisted of a thick and dense inner layer with transverse striations and a multi-lamellated outer layer formed by parallel dense internal bands alternating with lighter areas. The lamellae formed filamentous extensions of the wall. The sporocyst wall striation period was smaller than that observed in G. clupearum, which has a similar habitat. The dehiscence suture, characteristic of the genus, was present in the sporocysts of G. cruciata. The sporocysts were arranged in a symmetrical and characteristic cross shape. A large number of sporocysts with sporozoites were observed in direct contact with host liver cells. No macroscopic lesions were observed. In heavily infected fish, aggregations of oocysts were often enveloped in a 'yellow body' composed of amylopectin granules derived from the parasite and necrotic or aggregated host cells. Degenerating parasites were frequently observed in liver tissue. Host inflammatory cells were accumulated near some oocysts. The ultrastructure of the parasite, together with its strict host specificity, confirmed G. cruciata as a separate and valid species. PMID- 15752273 TI - Aquatic birnavirus induces apoptosis through activated caspase-8 and -3 in a zebrafish cell line. AB - In this study, the possible influence of temperature on infectious pancreatic necrosis virus (IPNV)-induced apoptosis in a zebrafish liver epithelium (ZLE) cell line was investigated. At a lower temperature (18 degrees C), there was expression of viral proteins VP2 and VP3 at 4 h post-infection (p.i.). At this time no expression was found in the high temperature group at 28 degrees C. The cell survival ratio was 52 and 18% at 24 and 48 h p.i., respectively, during IPNV infection at 18 degrees C. In addition, we assayed for apoptosis in IPNV-infected cells with terminal deoxynucleotidyl transferase (TdT)-mediated end labelling (TUNEL) of DNA at different dosages of virus. We found a ratio of apoptotic cells of 8 and 25% at 12 and 18 h p.i., respectively, in the multiplicity of infection (MOI) 1 group. The MOI 10 group had 20 and 45% apoptotic cells at 12 and 18 h, respectively. Furthermore, at 18 degrees C IPNV activated the caspase-8 and 3 from 1.5 to 2 times at 12 and 18 h p.i., respectively. Taken together, these findings suggest that successful virus replication occurs at the low temperature (18 degrees C) compared with the non-permissive temperature of 28 degrees C. Thus, IPNV replication is capable of activating caspase-8 and -3 and inducing host apoptosis. PMID- 15752274 TI - Aeromonas sobria, a causative agent of disease in farmed perch, Perca fluviatilis L. AB - Significant numbers of perch, Perca fluviatilis, raised on a pilot fish farm in Switzerland presented focal skin lesions on the lateral sides and fin rot. Mortality rates reached levels of up to 1% of the total fish on the farm per day. Virtually pure cultures of Aeromonas sobria were isolated from the liver, kidney, spleen and skin lesions of affected fish. Aeromonas sobria isolated from the farmed perch had a haemolytic effect on sheep and trout erythrocytes, autoaggregated, was cytotoxic for cultured fish cells and possessed genes involved in type III protein secretion. Experimental infection of naive perch with a single colony isolate of A. sobria from an affected farm fish resulted in the development of clinical signs identical to those seen on the farm. The results indicate that A. sobria can act as a primary pathogen of perch. PMID- 15752275 TI - Response of crayfish, Procambarus clarkii, haemocytes infected by white spot syndrome virus. AB - White spot syndrome virus (WSSV) is a serious pathogen of aquatic crustaceans. Little is known about its transmission in vivo and the immune reaction of its hosts. In this study, the circulating haemocytes of crayfish, Procambarus clarkii, infected by WSSV, and primary haemocyte cultures inoculated with WSSV, were collected and observed by transmission electron microscopy and light microscopy following in situ hybridization. In ultra-thin sections of infected haemocytes, the enveloped virions were seen to be phagocytosed in the cytoplasm and no viral particles were observed in the nuclei. In situ hybridization with WSSV-specific probes also demonstrated that there were no specific positive signals present in the haemocytes. Conversely, strong specific positive signals showed that WSSV replicated in the nuclei of gill cells. As a control, the lymphoid organ of shrimp, Penaeus monodon, infected by WSSV was examined by in situ hybridization which showed that WSSV did not replicate within the tubules of the lymphoid organ. In contrast to previous studies, it is concluded that neither shrimp nor crayfish haemocytes support WSSV replication. PMID- 15752276 TI - Effects of temperature, salinity, desiccation and chemical treatments on egg embryonation and hatching success of Benedenia seriolae (Monogenea: Capsalidae), a parasite of farmed Seriola spp. AB - The effects of temperature and salinity on the embryonation period and hatching success of eggs of Benedenia seriolae were investigated. Temperature strongly influenced embryonation period; eggs first hatched 5 days after laying at 28 degrees C and 16 days after laying at 14 degrees C. The relationship between temperature and embryonation period is described by quadratic regression equations for time to first and last hatching. Hatching success was >70% for B. seriolae eggs incubated at temperatures from 14 to 28 degrees C. However, no B. seriolae eggs embryonated and hatched at 30 degrees C and <2% of eggs hatched when incubated at 24 degrees C after transfer to 30 degrees C for 48 h. Embryonation period was similar for eggs incubated in sea water at 25, 30 and 35 per thousand salinity, but increased for eggs incubated at higher or lower salinities. When incubated at salinities ranging from 25 to 45 per thousand, more than 70% of B. seriolae eggs embryonated and hatched. Hatching success was lower at 20 and 50 per thousand salinity and few or no eggs hatched at 10 and 15 per thousand. Hatching of B. seriolae eggs can be prevented by desiccation for 3 min, by immersion in water at 50 degrees C for 30 s or by treatment with 25% ethanol for 3 min. PMID- 15752277 TI - Efficacy of furunculosis vaccines in turbot, Scophthalmus maximus (L.): evaluation of immersion, oral and injection delivery. AB - The commercial furunculosis vaccine Aquavac Furovac 5 and an autogenous vaccine, based on the challenge strain, induced immune protection in turbot, Scophthalmus maximus (L.), as shown in challenge tests 120 days post-immunization by injection (relative percentage of survival, RPS = 72-99%). This protective effect lasted for at least 6 months post-immunization at appreciable levels (RPS = 50-52%). Neither the autogenous vaccine nor the commercial vaccine was able to induce significant levels of protection against Aeromonas salmonicida in turbot when administered by immersion. Antibody levels were high or moderate in fish vaccinated by injection with the different vaccines and very low in fish vaccinated by immersion. The field results show that delivering an oral boost after the primary vaccination by injection did not enhance protection of turbot against furunculosis and that water-based (autogenous vaccine) and oil adjuvanted (Alpha Ject 1200) vaccines administered by injection conferred similar levels of protection (RPS > 80%) in turbot. PMID- 15752278 TI - Influence of feeding ratio and size on susceptibility to microsporidial gill disease caused by Loma salmonae in rainbow trout, Oncorhynchus mykiss (Walbaum). AB - Two trials were designed to quantify the effect of feeding ratio and fish size on the cohabitation transmission of Loma salmonae, the causative agent of microsporidial gill disease (MGD) in salmonids, Oncorhynchus spp. To evaluate the effect of feeding ratio on disease onset, groups of 45 rainbow trout, O. mykiss (Walbaum) (RBT), were fed daily at 1% (low), 2% (medium) or 4% (high) of the fish biomass in the tank. There were three tanks at each feeding level: two tanks were exposed to the pathogen and one was a control. For the second objective, 300 RBT were separated into seven tanks so that the weight classifications were small (17 23 g), medium (32-38 g) and large (57-63 g). Each size class was done in duplicate with one control tank containing medium-sized fish. Separately for each trial, on day 0 post-exposure (PE) five highly infectious RBT were added to each tank (not including the control tanks) to begin the cohabitation exposure period. Beginning on day 21 PE and continuing biweekly until days 70 and 77 PE for the feeding and fish size trials, respectively, each fish was evaluated for visible branchial xenomas to determine disease onset time. Using survival analysis, the survival curves for the low, medium and high feedings were not significantly different from each other. However, there were significant differences amongst the small, medium and large weight classes in the size trial. The median numbers of days to the development of branchial xenomas was 31, 38 and 42 for small, medium and large size fish, respectively. On any given day, a medium or large sized fish had a hazard ratio for developing branchial xenomas of 0.66 and 0.63, respectively, compared with a small fish. In addition to host species and host strain differences, fish size is now considered a host risk factor for the development of MGD. PMID- 15752279 TI - Experimental infection of turbot, Scophthalmus maximus (L.), by Aeromonas salmonicida subsp. achromogenes and evaluation of cross protection induced by a furunculosis vaccine. AB - Turbot was shown to be sensitive to injection challenges by Aeromonas salmonicida subsp. achromogenes (Asa). A systemic disease was induced and the bacterium was isolated from various internal organs. Histopathological changes involved haemorrhages, necrosis and degeneration in skin and muscle, haemorrhages and necrosis in kidney, degeneration in the heart muscle, and fusion of the secondary gill lamellae. A polyvalent commercial salmon vaccine, containing A. salmonicida subsp. salmonicida as one of five antigens, did not confer protection in turbot against an experimental Asa infection 13 weeks post-vaccination. Vaccination induced a significant antibody response against Asa cells but not against extracellular products of the bacterium. The results of the study indicate that Asa may be a potential threat to turbot farming and that the development of new turbot vaccines is needed. PMID- 15752280 TI - Pityriasis versicolor alba. AB - Pityriasis versicolor alba is a hypopigmented or depigmented variant of pityriasis versicolor characterized by maculous, partly pityriasiform, scaly depigmented lesions occurring particularly in seborrhoeic areas. Long-persisting hypopigmentation after healing of the pityriasis versicolor was first described by Gudden in 1853. Hypopigmentation and depigmentation were later differentiated as an independent variant of the disease. In 1848, Eichstedt recognized the pathogen-related character of pityriasis versicolor in its hyperpigmented form. Today it is generally accepted that the disease is caused by yeasts of the genus Malassezia, of which nine species are differentiated. It is controversial whether a single species is responsible for the disease. The pathogenesis of depigmentation has not been established. A screening effect by the scale layer as well as toxic effects on pigment synthesis by fungal metabolites have been discussed. With regard to the second mechanism, the newly discovered tryptophan derived metabolites of M. furfur might be significant. Evidence-based data concerning the therapy of pityriasis versicolor alba do not exist. According to current recommendations, pityriasis versicolor should be rapidly treated with antimycotics, followed by ultraviolet therapy to induce maturation of existent melanosomes and accelerate repigmentation. However, depigmented lesions are difficult to improve by ultraviolet therapy. PMID- 15752281 TI - Burnout and job satisfaction comparing healthcare staff of a dermatological hospital and a general hospital. AB - BACKGROUND: Psychological distress among healthcare professionals can have negative effects on the well-being of the professionals and also on the quality of care they provide to patients. OBJECTIVES: To evaluate burnout and job satisfaction of dermatologists and nurses working with dermatological patients compared with physicians and nurses of other specialties. METHODS: A self completed anonymous questionnaire was distributed to the personnel of two hospitals in Rome, Italy: a dermatological hospital (IDI) and a general hospital (GH), belonging to the same non-profit organization. Standardized instruments were used to assess burnout (Maslach Burnout Inventory) and job satisfaction. Multiple logistic regression was used to examine the association between burnout and working in dermatology vs. other specialties, job satisfaction, years of employment and respondents' sex and age. RESULTS: We distributed 929 questionnaires to clinical and non-clinical staff of IDI and 494 questionnaires to the GH staff (response rates: 53% at IDI and 50% at the GH). Among respondents there were 67 physicians and 59 nurses at IDI and 70 physicians and 148 nurses at the GH. Subsequent analyses refer only to this clinical subsample. Factor analysis showed that among physicians and nurses the two main factors explaining job satisfaction were respondents 'satisfaction with the management of their unit' and 'opportunities for personal growth'. Among nurses the likelihood of burnout decreased significantly with higher levels of job satisfaction [odds ratio (OR) = 0.78; 95% confidence interval (95% CI) 0.7-0.9] and working in dermatology compared with other specialties (OR = 0.46; 95% CI 0.2-0.9). Among physicians a lower likelihood of burnout was associated with job satisfaction (OR = 0.66; 95% CI 0.5-0.8) and older age (OR = 0.28; 95% CI 0.1-0.8). CONCLUSIONS: Among both physicians and nurses, job satisfaction was associated with a lower likelihood of burnout, independently of clinical specialty and other factors. Burnout was similar for dermatologists and other specialists. Nurses of the GH compared with those working in dermatology had a higher probability of burnout and were significantly less satisfied with the management of their units and with opportunities for personal growth. PMID- 15752282 TI - Intravenous infusion for the treatment of diabetic and ischaemic non-healing pedal ulcers. AB - Diabetic and ischaemic non-healing pedal ulcers have a tendency for chronicity and increased chances of infection, which may threaten the viability of the foot. Systemic administration of therapeutic agents may be insufficient in these cases. We have assessed the role of retrograde venous perfusion (RVP) for the treatment of nine diabetic and 10 ischaemic non-healing pedal ulcers. Agents used were soda bicarbonate, heparin, lignocaine, gentamicin and pentoxiphylline. Five of nine diabetic non-healing ulcers showed complete healing and the remaining four improved. The complete recovery in the cases of diabetic ulcer occurred in 10-24 days (mean 16 days), while ischaemic ulcers took 10-14 days for complete recovery (mean 13.6 days). There was a reduction of rest pain in all 10 patients with ischaemic disease; five patients showed complete healing of ulcers, and the other five improved significantly. In two patients, pre-gangrene changes were reversed. RVP is a useful adjunct to conservative or surgical treatment of non-healing pedal ulcers. Its main impact was in improving ischaemia and promoting healing. PMID- 15752283 TI - Effects and side-effects of spironolactone therapy in women with acne. AB - BACKGROUND AND AIMS: Androgen hormones play an important role in the pathogenesis of acne. Despite the demonstrated effects, spironolactone, an androgen receptor blocker, is not commonly used to treat acne. We planned an open-labelled, prospective study to evaluate the effects and side-effects of spironolactone therapy in women with acne. MATERIALS AND METHODS: Thirty-five consecutive patients with acne were treated with spironolactone 100 mg/day, 16 days each month for 3 months. The patients were divided according to the clinical severity of the lesions as having mild, moderate and severe acne. Serum total testosterone and dehydroepiandrosterone sulfate (DHEAS) levels were measured before and after treatment. Lesion numbers and hormone levels before and after treatment were compared with one-sampled t-test. RESULTS: The mean age of the patients was 21.4 +/- 3.5 years. Two patients discontinued the study due to side-effects. Five patients were lost in the follow-up. Clinically significant improvement was noted in 24 patients (85.71%). No response was seen in four patients. All of the nonresponding patients had received previous unsuccessful therapies. Mean number of lesions and mean DHEAS levels of the 24 patients with clinical improvement decreased significantly after treatment (P < 0.01 and P < 0.05, respectively). There was no change in the mean total testosterone levels before and after treatment (P > 0.05). CONCLUSION: Spironolactone is a safe and effective medication for women with acne vulgaris. Although its side-effects seem to be high, they are in the majority of cases not a reason to stop treatment. PMID- 15752284 TI - Evaluation of nitrosative and oxidative stress in Behcet disease. AB - Plasma nitrotyrosine and nitrite/nitrate levels as markers of nitrosative stress and plasma malondialdehyde (MDA) and protein carbonyl as markers of oxidative stress were determined in patients with Behcet disease (BD). To evaluate the balance between oxidant and antioxidant systems in these patients, we measured erythrocyte lysate CuZn superoxide dismutase (CuZn SOD) activity, plasma sulfhydryl (SH) values and total antioxidant activity. We also determined levels of plasma C-reactive protein (CRP), a key marker of inflammation, and compared them with those of healthy subjects. We found plasma nitrotyrosine levels of BD patients to be increased, indicating that nitrosative stress may occur in these patients. Plasma MDA and CRP levels in BD patients were found to be significantly higher than those in control group. However, plasma SH levels were decreased. No changes were observed in the other measured parameters of the patient group compared with the controls. These data suggest the possible involvement of nitric oxide (NO) together with reactive oxygen substances (ROS) in the pathogenesis of BD. PMID- 15752285 TI - Oral itraconazole for the treatment of seborrhoeic dermatitis: an open, noncomparative trial. AB - BACKGROUND: Seborrhoeic dermatitis is an inflammatory cutaneous disorder in which the colonization of the affected area by Malassezia has been proved to play a key role. OBJECTIVE: To perform a noncomparative open clinical study with oral itraconazole capsule (200 mg/day x 7 days) and consecutive usage 200 mg/day for the first 2 days of the following 2 months in patients with seborrhoeic dermatitis. METHODS: Twenty-nine patients were enrolled to determine the efficacy and safety of oral itraconazole. The patients were evaluated according to itching, burning, erythema, desquamation and seborrhoea, each scored on a 0-4 scale on days 15 (T15), 30 (T30), 60 (T 60) and 90 (T90). Itraconazole capsule 100 mg was given twice a day for 1 week and then, after a 3-week interval, patients used itraconazole capsule 200 mg/day for the first 2 days of the following 2 months. The clinical response was graded as markedly effective, effective, moderate or ineffective. RESULTS: A clinical improvement (evaluated as markedly effective or effective) was observed in 23 patients (83%) at T15, 21 (76%) at T30, 20 (72%) at T60 and 17 (61%) at T90. At baseline, the mean +/- SD total clinical scores were 10.44 +/- 2.45, 1.98 +/- 0.5, 2.97 +/- 1.12, 3.15 +/- 1.74 and 3.30 +/- 1.90 at T0, T15, T30, T60 and T90, respectively. Compared with baseline values, itraconazole capsule significantly reduced the mean +/- SD total score as well as individual erythema and desquamation (Wilcoxon's signed test-two tailed) (P < 0.0001). No drug-related systemic adverse event was observed during the study. CONCLUSIONS: Seborrhoeic dermatitis shows marked reduction in inflammation when treated with itraconazole. The anti-inflammatory activity of oral itraconazole and efficacy on Malessezia suggests that itraconazole capsule will be first oral treatment option in future in severe seborrhoeic dermatitis. PMID- 15752286 TI - Dithranol irritation in psoriasis treatment: a study of 68 inpatients. AB - The irritant response of perilesional skin is a serious limitation of dithranol therapy in psoriasis. No data are available on the actual prevalence and severity of irritation during 24-h dithranol treatment in an inpatient setting. Using a retrospective analysis of 68 patients with psoriasis visiting our inpatient department for dithranol treatment, the occurrence of dithranol irritation was studied. We found a relatively high frequency of dithranol irritation. Furthermore, most irritation occurs at the start of the therapy with relatively low concentrations. PMID- 15752287 TI - Immunophenotypic analysis of the p53 gene in non-melanoma skin cancer and correlation with apoptosis and cell proliferation. AB - BACKGROUND: Sunlight precipitates a series of genetic events that lead to the development of skin cancers such as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). The p53 tumour suppressor gene, which plays a pivotal role in cell division and apoptosis, is frequently found mutated in sunlight-induced skin tumours. OBJECTIVE: To investigate the immunoreactivity of the p53 gene in non melanoma skin cancers and to correlate its expression with apoptotic and cell proliferation markers. METHODS: We analysed 35 non-melanoma tumours including 19 BCCs and 16 SCCs from sun-exposed skin areas. p53 protein expression was studied immunohistochemically using the DO7 monoclonal antibody against wild-type and mutant p53 forms. The percentage of p53-immunopositive nuclei was measured by image analysis. Cell proliferation and apoptosis were also assessed by image analysis following Ki-67 immunostaining and application of the TUNEL method on paraffin sections, respectively. RESULTS: The percentage of p53-expressing cells varied from 3.5 to 90 in BCCs (median value 54.4%) and from 3.7 to 94 in SCCs (median value 40.3%). The mean value of Ki-67-positive cells was comparable in both groups of tumours with a mean value of 40.6% in BCCs and 34.6% in SCCs. Conversely, the TUNEL assay showed sporadic staining of apoptotic cells within the tumours with a mean value of 1.12% in BCCs and 1.8% in SCCs. p53 protein expression was correlated positively with cell proliferation (r = 0.75, P = 0.000001) and negatively with apoptosis (r = -0.23, P = 0.05). CONCLUSION: p53 immunoreactivity was high in the majority of the skin carcinomas examined and correlated positively with cell proliferation and negatively with apoptosis. The p53 protein overexpression appears to be related to an inactivated protein resulting from mutations of the p53 gene or other unclear molecular mechanisms. PMID- 15752288 TI - Ultraviolet A dosimetry in photopatch test centres in Europe. AB - OBJECTIVE: The aim of the study was to investigate the state of ultraviolet (UV) A dosimetry in photopatch testing centres in Europe. BACKGROUND: It has been acknowledged that a widespread disparity exists in the way photopatch testing is carried out in different countries throughout Europe. This prompted the formation of a European taskforce that developed a consensus methodology. One issue that was not addressed directly was that of UVA dosimetry, which clearly impacts on the consistency of the clinical results. STUDY DESIGN: Ten UV meters from photopatch test centres throughout Europe were set up at a reproducible position from a photopatch test light source containing UVA fluorescent lamps and the irradiances were noted from each meter. One meter served as a reference meter with calibration carried out in an optical laboratory traceable to the National Physical Laboratory. RESULTS: The mean of all the measurements was 5.74 mW/cm(2) with a SD of 0.36 mW/cm(2). Expanded uncertainty at 95% confidence level was 12%. CONCLUSIONS: It is desirable that meter calibration should have an accuracy of the order of +/-10%. The variation in recorded values among the 10 meters investigated was within acceptable limits. The meter must be calibrated using a similar light source to that used in the phototest equipment, in this case a UVA fluorescence lamp. PMID- 15752289 TI - The effects of foot disease on quality of life: results of the Achilles Project. AB - BACKGROUND: Foot diseases have a high prevalence in the general population, but their impact on quality of life has not been assessed in large-scale studies. The Achilles Project surveyed foot disease in patients visiting their primary care physician or dermatologist. METHODS: A total of 43 593 patients were asked about the impact of their condition on their quality of life: pain, discomfort in walking, limitations in daily activities, and embarrassment. RESULTS: Overall, 52.5% of patients had some aspect of their quality of life affected by their foot disease. More specifically, 30.7% of patients experienced pain, 40.3% had discomfort in walking, 19.6% had their daily activities limited, and 27.3% were embarrassed. The survey indicated a larger impact of foot disease on the quality of life of women vs. men in all categories, except for daily activities. Similarly, the elderly (> or = 65 years) were more affected by their foot disease, although they suffered no more embarrassment than other age groups. Participation in sports seemed to lower the proportion of patients who had their quality of life adversely affected. Non-fungal foot diseases, particularly ulcer and gangrene, are more likely to cause pain, discomfort in walking and limit daily activities, than fungal diseases. CONCLUSIONS: In general, non-fungal foot diseases caused pain, discomfort in walking and limitations in daily activities in more patients than fungal foot diseases, but a higher proportion of patients with fungal foot diseases were embarrassed by their condition than patients with non-fungal foot diseases. The study found that the impact of foot disease on quality of life may be greater than previously suggested. Given that effective treatments are available, routine examination of patients' feet by dermatologists and primary care physicians may help to reduce the burden of these foot conditions. PMID- 15752290 TI - Methotrexate for psoriasis: current European practice. A postal survey. AB - The aim of this survey was to determine current practice throughout Europe regarding use and monitoring of methotrexate therapy for psoriasis. A structured questionnaire with questions on methotrexate prescribing and monitoring was mailed to 150 dermatologists in 32 European countries in June 2002. Dermatologists' names were chosen at random from the 2001 European Academy of Dermatology and Venereology membership directory. A reply was received from 69 dermatologists of whom 59 prescribed methotrexate regularly. In those patients receiving systemic treatment for psoriasis, methotrexate was the most widely used drug (4.4 patients of every 10) followed by acitretin (3.2) and cyclosporin (1.6). Myelosuppression was the commonest reported fatal side-effect of methotrexate (eight of a total of 10 cases). None of the respondents routinely requested a baseline liver biopsy before starting methotrexate treatment and of every 10 of their patients on long-term methotrexate it was estimated that less than two had had a liver biopsy some time during treatment. Serum measurement of the amino-terminal peptide of type III procollagen (PIIINP) was used routinely to detect liver fibrosis by 12 (20%) of the 59 respondents who regularly prescribed methotrexate. This survey demonstrates that, despite the advent of new therapies, methotrexate retains a central role in the treatment of severe psoriasis in Europe. The responses demonstrated variation in, among others, the methotrexate dose regimen utilized and the use of folate supplementation, confirming the need for randomized controlled studies to address these issues. PMID- 15752291 TI - Successful UVA1 phototherapy in a patient with scleredema adultorum. AB - Scleredema adultorum is a rare connective tissue disorder of unknown cause. Both bath-PUVA and cream-PUVA therapy were reported to be effective. We describe a patient with scleredema adultorum who showed a striking clinical improvement with a medium-dose UVA1 phototherapy (single dose, 50 J/cm(2); 35 treatments). PMID- 15752292 TI - Liver failure and transplantation after itraconazole treatment for toenail onychomycosis. AB - Three weeks after completing a 4-pulse course of itraconazole for toenail onychomycosis, a 25-year-old woman patient developed severe liver crisis and required an emergency liver transplant. We report the case and discuss the use of itraconazole in onychomycosis and dermatomycoses. PMID- 15752293 TI - Multinucleate cell angiohistiocytoma: a new case report. AB - Multinucleate cell angiohistiocytoma (MCAH) was first described by Smith and Wilson-Jones in 1985. It is an uncommon entity but probably underdiagnosed because of lack of recognition by clinicians and pathologists. We report a 47 year-old man with asymptomatic grouped violaceous papules on the dorsum of the hands for 3 years. The histopathological and immunopathological features of our case revealed characteristics of MCAH similar to the initial description of Smith and Wilson-Jones and other reports. PMID- 15752295 TI - Reactive angioendotheliomatosis: is it 'intravascular histiocytosis'? AB - We report the case of a 68-year-old female with reactive angioendotheliomatosis (RAE). This case highlights the benign course of this condition and suggests that this entity might be an intravascular histiocytosis. PMID- 15752294 TI - Pseudoxanthoma elasticum and nephrolithiasis. AB - We report the case of a 42-year-old man with pseudoxanthoma elasticum (PXE) and recurrent bilateral nephrolithiasis. Diagnosis of PXE was made by yellow papules on the neck and ophthalmologic angioid streaks. This diagnosis was confirmed by a skin biopsy (Von Kossa stain) and by genotyping analysis of ABCC6 (homozygous mutation R1138Q). Nephrolithiasis was recurrent and biological investigations showed hypophosphoraemia, hyperphosphaturia, hypercalciuria, normocalcaemia, normal serum parathyroid hormone value, high 1,25-dihydroxy vitamin D value and a renal calcium oxalate stone. ABCC6 encodes for MRP6, a multidrug resistant protein strongly expressed in the liver and kidney. The substrates of the MRP6 remain unknown. As PXE is characterized by calcification of elastic fibres and this patient presents important phosphocalcic anomalies, we discuss the possible implications of MRP6 in the phosphocalcic metabolism. PMID- 15752296 TI - Autodestruction of penis due to carcinoma. AB - Four unusual cases of autodestruction of the penis are described with their aetiology, diagnosis and management. Causes for delay in diagnosis and therapy, prevention and early recognition of this condition are emphasized. PMID- 15752297 TI - Dermatitis artefacta: unusual appearance in an older woman. AB - Dermatitis artefacta is a rare and difficult condition for diagnosis and treatment, with the highest incidence of onset in late adolescence to early adult life. Most patients are young women who have a personality disorder; borderline features are common and the patient's denial of psychological distress makes management and treatment difficult. Patients use a variety of means to cause the skin changes. Clinical presentation of the skin lesions does not conform to those of known dermatoses and are located on easily reached parts of the skin. We report an unusual case of a 72-year-old woman with symmetrical changes under the breasts and in the right inguinal region. The lesions were composed partly of haemorrhagic round lesions and partly of scars. A skin biopsy was taken and consultations with the psychiatrist, internist and the patient's family led to the diagnosis of self-induced dermatitis. The skin lesions were covered by occlusion techniques and the lesions improved very rapidly. The patient was discharged from the hospital under psychiatric and family care. PMID- 15752298 TI - Eosinophilic cellulitis of papulonodular presentation (Wells' syndrome). AB - Eosinophilic cellulitis (Wells' syndrome) is a rare condition of unknown aetiopathogenesis and is characterized by erythemal plaques and a histological picture of eosinophilic infiltration of the dermis with 'flame' figures. Here we describe a patient with the papulonodular variant of eosinophilic cellulitis associated with an unusual hepatic dysfunction. PMID- 15752299 TI - Hypereosinophilic dermatitis-like erythema annulare centrifugum in a patient with chronic lymphocytic leukaemia. AB - A 65-year-old male presented with a history of disseminated erythema annulare centrifugum lesions and a high peripheral blood eosinophilia (55%). Histopathology of a lesion revealed a superficial and deep perivascular and interstitial inflammatory eosinophilic infiltrate. The bone marrow aspirate showed a heavy eosinophilic infiltrate. There was no evidence of any other systemic involvement except for bone marrow eosinophilia. The patient had a 6 year history of chronic lymphocytic leukaemia. The disease is in complete remission for the last 3 years. The clinical and histopathological findings in the reported patient correspond to the diagnosis of hypereosinophilic dermatitis. Topical treatment with corticosteroid creams was successful. The cutaneous lesions gradually resolved in 8 weeks, within 7 months the peripheral blood eosinophilia slowly declined from 55% to 7%. PMID- 15752300 TI - Bowen's carcinoma of the scrotal skin associated with human papillomavirus type 82. AB - We have previously cloned human papillomavirus type 82 (HPV-82) from a vaginal intraepithelial neoplasia, but it is not known whether HPV-82 can induce a cutaneous lesion. A large erosive nodule developed on the scrotum of a 50-year old Japanese patient. Histopathologically, the lesion was composed of two distinct parts; one part showing changes characteristic of Bowen's disease in the epidermis, and the other showing elongated rete ridges and proliferation of atypical basaloid cells in the dermis. These parts were partially connected, giving the diagnosis of Bowen's carcinoma. Immunohistochemically, HPV capsid antigen was detected only in the nuclei of a few cells on the upper part of the epidermis. HPV-82 was identified in the lesion by blot hybridization and viral DNA was demonstrated in the lesion by in situ hybridization. HPV-82 has tropism for both the skin and the genital regions. PMID- 15752302 TI - Increased activity of factor VIII coagulant associated with venous ulcer in a patient with Klinefelter's syndrome. AB - Klinefelter's syndrome is the most frequent major abnormality of sexual differentiation in men with two or more X chromosomes. Recurrent venous ulcers as a result of a post-thrombotic syndrome are a well known symptom in patients with Klinefelter's syndrome. Until now the underlying pathomechanisms are not completely understood. Platelet hyperaggregability, factor V Leiden mutation and abnormalities in fibrinolysis were implicated as possible contributing factors. Here we describe the detection of an increased activity of factor VIII coagulant (factor VIII:C). This is the first case report on increased factor VIII:C activity associated with venous ulcers in a patient with Klinefelter's syndrome. Elevated factor VIII plasma levels are gradually accepted to be associated with an increased risk for venous thromboembolism. Therefore, we discuss that the examination of factor VIII:C may help in clarifying individual thromboembolic risks, especially in patients with Klinefelter's syndrome. PMID- 15752301 TI - Management of erosive lichen planus with topical tacrolimus and recurrence secondary to metoprolol. AB - Metoprolol, a widely prescribed beta-adrenergic receptor blocker, has occasionally been associated with a diversity of cutaneous reactions. We present a 79-year-old male patient with erosive lichen planus (LP) on the feet and hands who was successfully treated with topical tacrolimus. Six months after the lesions had been cured the patient received the beta-receptor blocker metoprolol for the treatment of hypertonus. Within only 2 weeks of metoprolol intake the erosive lesions on the palms and feet recurred. After discontinuation of the drug and repetitive topical treatment with tacrolimus a complete remission of the lesions could be achieved. The recurrence of erosive LP probably secondary to metoprolol and the therapeutic success of topical tacrolimus in the treatment of LP are discussed. PMID- 15752303 TI - Successful treatment of a leg ulcer occurring in a rheumatoid arthritis patient under leflunomide therapy. AB - OBJECTIVE: We report the case of a leg ulcer in a rheumatoid arthritis (RA) patient under treatment with leflunomide, discuss the influence of the drug on the aetiopathogenesis of the ulcer and describe its successful treatment. CASE SUMMARY: A 68-year-old woman with a 12-year history of RA developed a leg ulcer after 4 months of leflunomide treatment. Other ulcerogenic factors were ruled out. There were some clinical hints for rheumatoid vasculitis. The ulcer was resistant to ambulant conservative phase adapted wound bed preparation and a split skin transplantation failed. After omission of leflunomide and washout procedure with cholestyramine a second split skin transplantation resulted in complete healing. DISCUSSION: Leflunomide inhibits the division of activated T cells and thus inhibits among others the production of proinflammatory cytokines and the adhesion of cells to the endothelium. These mechanisms may partly explain the possible influence of leflunomide on the perpetuation of the ulcer. Until now, occurrence of vasculitis and leg ulcers has been described in one case each for the novel immunomodulator leflunomide. No successful treatment of a leg ulcer under leflunomide has been described yet. Omission of leflunomide and a washout treatment in our case led to a complete healing. This may indicate a critical role of leflunomide in the maintenance of this slow healing ulcer. CONCLUSIONS: An association between leflunomide intake, occurrence of leg ulcers in RA patients and delayed wound healing should be considered. PMID- 15752304 TI - Delayed onset of warts over tattoo mark provoked by sunburn. AB - Multiple warts in a 32-year-old-man are reported that developed after tattooing and remaining exclusively confined to that area. The tattooing was done 2.5 years earlier by a professional tattoo artist. It was previously a lesion-free tattoo, but when damaged by sunburn developed multiple skin warts. The ability of a latent virus to induce warts after cutaneous ultraviolet exposure was discussed. PMID- 15752305 TI - Tacrolimus ointment for the treatment of severe facial plaque psoriasis. AB - Recently, tacrolimus ointment has proved to be effective and well tolerated in patients with facial psoriasis. A few months ago we had the opportunity to treat a patient with tacrolimus ointment who had severe and recalcitrant plaque psoriasis of the face. This present case illustrates the impressive improvement of facial plaque psoriasis following 5 months of treatment with tacrolimus 0.1% ointment twice a day. Significant improvement of facial plaque psoriasis was seen after 1 month and complete clearance after 5 months of therapy. Based on the available literature and illustrated by the present case we may conclude that tacrolimus ointment 0.1% can be recommended as a first-line treatment for facial psoriasis. PMID- 15752307 TI - Acquired periungal arteriovenous tumour (cirsoid aneurysm). PMID- 15752306 TI - Livedo reticularis associated with interferon alpha therapy in two melanoma patients. AB - We report two patients who developed intense livedo reticularis clearly related to the administration of interferon alpha 2b as an adjuvant therapy for melanoma. Histological studies showed scattered perivascular infiltrates without vasculitis. Laboratory tests excluded any underlying condition. Resolution of the symptoms was observed in both patients when interferon alpha was withdrawn. These cases highlight the occurrence of livedo reticularis as an uncommon side-effect of interferon alpha treatment. PMID- 15752308 TI - Nodular syphilis mimicking histoid leprosy. PMID- 15752309 TI - Dermatomyofibroma in a male child. PMID- 15752310 TI - Bilateral callosities, plantar ulcers and peripheral neuropathy in a 15-year-old nondiabetic boy--think of neuroborreliosis. PMID- 15752311 TI - Lupus mastitis: a 2-year history of a single localization of lupus erythematosus mimicking breast carcinoma. PMID- 15752312 TI - A case of lichen amyloidosis treated with pulsed dye laser. PMID- 15752314 TI - Tubercular lymphadenitis with lymphangiectases of the vulva. PMID- 15752313 TI - Milia en plaque. PMID- 15752315 TI - Lichen planus in children and adolescents: a report of eight cases. PMID- 15752316 TI - Darier's disease (keratosis follicularis): gynaecomastia unique hitherto unreported association. PMID- 15752317 TI - Improving nursing services through higher education. PMID- 15752318 TI - Assessing the postoperative patient: philosophy, knowledge and theory. AB - The assessment of the postoperative patient is critical in identifying the patient's current condition and to monitor any complications that might occur. However, many nurses fail to realize the implications that philosophical paradigms, knowledge and theoretical concepts bring to the assessment practice and the manner in which assessment is conducted; in particular, within 24 h after surgery. This article seeks to explore the usefulness of philosophy, knowledge and theory as they relate to nursing assessment of the postoperative patient within the initial 24 h after surgery. It is the duty of nurses to select and effectively apply appropriate philosophical dimensions, knowledge and theories in practice and critically evaluate these areas with respect to assessment of the postoperative patient, thus aiming for a holistic assessment of the patient. PMID- 15752319 TI - Making choices: how older people living in independent living units decide to enter the acute care system. AB - Older people living in independent living units make choices about where they live and the degree of support required to maximize their health and well-being. This can include when to enter the acute care system. Using a multimethod, multistage qualitative approach, this study aims to explore and describe the decision-making process of older people living in independent living units to enter the acute care system. Based on the findings, recommendations are provided which can ensure that older people do not enter acute care facilities until they need to, or if they do need to, they can access the care they require and leave with the best possible chance of not re-entering unnecessarily. The findings highlight that it is not enough to focus on the older person at the point of entry into the acute care system so the recommendations aim to assist in the development of best-practice initiatives for older people living in independent living units. PMID- 15752320 TI - Are patients with or at risk of pressure ulcers allocated appropriate prevention measures? AB - The aims of the study were to investigate the risk for and prevalence of pressure ulcers in different medical care groups, to discover if patients at risk for or with pressure ulcers are allocated appropriate pressure ulcer preventions and to investigate which variables are associated with appropriate pressure ulcer preventions. A cross-sectional survey design was used and followed the methodology developed by the European Pressure Ulcer Advisory Panel. A total of 612 patients participated in the study. The prevalence of pressure ulcers was greatest in geriatric care, followed by intensive care, acute care and neurological care. The majority of patients at risk for or with pressure ulcers did not receive appropriate preventative measures, either while they were in bed or in a chair. Significant variables associated with appropriate preventions in bed were intensive care, geriatric care, a low Braden score, a low score in the subscale activity and a long hospital stay. PMID- 15752321 TI - Conducting client supervision in community health care. AB - The aim of the present study was to examine registered public health nurses' views and reported actions related to client supervision in community health care. Transcript-based, qualitative and interpretative content analysis was used to analyse the findings from six focus group interviews. The main findings represented the following themes: the components of the assessment basis related to choosing supervision as an intervention strategy, the strategies related to conducting supervision and the meaning of supervision as an intentional act. In conclusion, the registered public health nurses' reported model for conducting supervision seems to correspond with principles in the new strategies for health promotion initiated by the World Health Organization. Further research is needed to explore what the nurses really do when conducting supervision in order to provide extended knowledge about their model. PMID- 15752322 TI - The care encounter: pondering caring, honest communication and control. AB - The proposition presented in this paper is that caring, underpinned by beneficence and non-maleficence, assumes that nurses will make determinations about another's best interests and act accordingly. With some emphasis on high level (nursing home) aged care, it is the author's contention that nurses give priority to communication that controls the care encounter rather than acting in another's best interests. This view is supported in the literature, which suggests that nurses perceive honesty and information-sharing as less important in caring than do patients, and nurses' perception that telling another the truth is harmful and, therefore, contravenes the duties of beneficence and non maleficence. PMID- 15752323 TI - Operating theatre nurses: emotional labour and the hostess role. AB - Emotional labour has been established as a significant factor in nursing work, although no studies have been done looking at emotional labour specifically in an operating theatre nursing context. Theatre staff (17 nurses and three Operating Department Practitioners (technicians) were observed in practice over a period of nine months by one of the authors. Each of the staff was subsequently interviewed. The transcriptions of the observation fieldwork notes and the semistructured interviews were analysed for themes and content. The (predominantly female) nurses perceived that one of their responsibilities was 'looking after the surgeons'. We have described this as the 'hostess' role. This role consisted of two major areas of activity: 'keeping the surgeons happy' and 'not upsetting the surgeons'. Examples are given of how this was accomplished through talk and actions. The (predominantly male) operating department practitioners did not see this as part of their work. This 'hostess' role is a kind of emotional labour, but performed with coworkers rather than patients. Like other forms of emotional labour, it is strongly gendered. The emotional labour performed by the theatre nurses was necessary to maintain what has been called elsewhere the 'sentimental order'. PMID- 15752324 TI - Tempe fermentation, innovation and functionality: update into the third millenium. PMID- 15752325 TI - Maltose utilization in Enterococcus faecalis. AB - AIMS: The aim of this research was to characterize the metabolic pathway for maltose utilization in Enterococcus faecalis. METHODS AND RESULTS: Screening a library of Enterococcus faecalis insertional mutants allowed the isolation of mutants affected in maltose utilization. Genetic analysis of the insertion loci revealed insertions in neighbour genes encoding an EII component of a phosphotransferase system (PTS) transporter (malT) and a maltose phosphorylase homologue (malP). The malP gene forms part of an operon which also includes genes encoding a phosphoglucomutase (malB), a mutarotase (aldose 1-epimerase) (malM) and a transcriptional regulator (malR). Analysis of (14)C-labelled carbohydrates uptake revealed that more than 97% of maltose enters the cells by the PTS transporter MalT. CONCLUSIONS: Both experimental data and genetic organization of the malPBMR operon strongly suggest that in Enterococcus faecalis, maltose enters using a PTS, leaving maltose-6-phosphate inside the cells which is hydrolysed by a maltose phosphate phosphorylase (MalP). SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes a new pathway for maltose utilization in lactic acid bacteria. PMID- 15752326 TI - Survival of E. coli O157:H7 in organic wastes destined for land application. AB - AIM: To determine the persistence of Escherichia coli O157 in contrasting organic wastes spread to land and to assess the potential environmental risk associated with the disposal of these wastes to land. METHODS AND RESULTS: Twenty-seven organic wastes originating from slaughterhouses, wastewater treatment plants (raw and treated sewage), creameries and farms (bovine slurry), were inoculated with E. coli O157:H7 and incubated at 10 degrees C. Although pathogen numbers gradually declined in all the wastes, albeit at different rates even in the same waste type, E. coli O157:H7 was still viable in 77% of organic wastes tested after 2 months. CONCLUSIONS: Long-term storage of organic wastes led to a significant and gradual decline in E. coli O157:H7 numbers. Consequently, storage may be a useful means of reducing the pathogen load of wastes destined for land application. However, in most cases, long-term storage cannot be expected to completely eliminate E. coli O157:H7 from waste. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate that current legislation may be insufficient to protect the environment from E. coli O157:H7 contamination from untreated wastes spread to land. PMID- 15752327 TI - Improvement of bioinsecticides production through adaptation of Bacillus thuringiensis cells to heat treatment and NaCl addition. AB - AIMS: The present work aimed to increase yields of delta-endotoxin production through adaptation of Bacillus thuringiensis cells to heat shock and sodium chloride and to investigate their involvements in bioinsecticides production improvement. METHODS AND RESULTS: Growing B. thuringiensis cells were heat treated after different incubation times to study the response of the adaptative surviving cells in terms of delta-endotoxin synthesis. Similarly, adaptation of B. thuringiensis cells to sodium chloride was investigated. Adaptation to combined stressors was also evaluated. When applied separately in the glucose based medium, 20-min heat treatment of 6-h-old cultures and addition of 7 g l(-1) NaCl at the beginning of the incubation gave respectively 38 and 27% delta endotoxin production improvements. Heat shock improved toxin synthesis yields, while NaCl addition improved delta-endotoxin production by increasing the spore titres without significant effect on toxin synthesis yields. Cumulative improvements (66%) were obtained by combination of the two stressors at the conditions previously established for each one. Interestingly, when the similar approach was conducted by using the large scale production medium based on gruel and fish meal, 17, 8 and 29% delta-endotoxin production improvements were respectively, obtained with heat shock, NaCl and combined stressors. CONCLUSIONS: Heat treatment of vegetative B. thuringiensis cells and NaCl addition to the culture media improved bioinsecticides production. Heat treatment increased toxin synthesis yields, while addition of NaCl increased biomass production yields. Cumulative improvements of 66 and 29% were obtained in glucose and economic production media, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Overproduction of bioinsecticides by B. thuringiensis could be obtained by the combination of heat treatment of vegetative cells and addition of NaCl to the culture medium. This should contribute to a significant reduction of the cost of B. thuringiensis bioinsecticides production and utilization, and also manage for higher toxin content in the bioinsecticides, which is very interesting from a practical point of view because fewer spores would be disseminated into the ecosystem. PMID- 15752328 TI - Cloning and characterization of aspartate-beta-semialdehyde dehydrogenase from Mycobacterium tuberculosis H37 Rv. AB - AIMS: To clone and characterize the aspartate-beta-semialdehyde dehydrogenase of Mycobacterium tuberculosis H37Rv. METHODS AND RESULTS: The asd gene of M. tuberculosis H37Rv was cloned in pGEM-T Easy vector, subcloned in expression vector pQE30 having a T5 promoter, and overexpressed in Escherichia coli. The ASD enzyme was expressed to levels of 40% but was found to be inactive. Functional ASD was obtained by altering induction and growth conditions and the enzyme was purified to near homogeneity using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. The K(m) and V(max) values for the three substrates L-ASA, NADP and Pi, the turnover number and specific activity of the enzyme were determined. CONCLUSIONS: Functional ASD enzyme of M. tuberculosis was obtained by gene cloning and protein purification using affinity chromatography. The K(cat) and specific activity of the enzyme were 8.49 s(-1) and 13.4 micromol min(-1) microg( 1) respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The ASD enzyme is a validated drug target. We characterized this enzyme from M. tuberculosis and future work would focus on deducing the three-dimensional structure of the enzyme and design of inhibitors, which could be used as drugs against TB. PMID- 15752329 TI - Aspergillus carbonarius growth and ochratoxin A production on a synthetic grape medium in relation to environmental factors. AB - AIMS: The effects of water activity (0.90-0.99 a(w)), temperature (15-37 degrees C), and their interaction on growth and ochratoxin A (OTA) production by eight isolates of Aspergillus carbonarius were investigated on synthetic nutrient medium (SNM) with composition similar to grapes. METHODS AND RESULTS: Growth data were modelled by an multiple linear regression and response surface models were obtained. Aspergillus carbonarius grew much faster at 30 degrees C than at the other temperature levels tested and its growth rate increased with increasing a(w), maximum growth rate being between 0.95 and 0.99 a(w). In general, isolates grew faster at 35-37 degrees C than at 20 degrees C, although no significant differences were found between these temperatures. OTA accumulation was also favoured by high a(w) levels, and although it was observed in the whole range of temperatures, maximum amounts were detected at 20 degrees C. No OTA was found at the most unfavourable growth conditions. CONCLUSIONS: Optimum a(w) level for growth seems to correspond with optimum for OTA production, meanwhile the most propitious temperature for the toxin production was below the best one for growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Prediction of A. carbonarius growth would allow estimating their presence and therefore, the OTA production, as it was found that conditions for the toxin production were more limited than those permitting growth. PMID- 15752330 TI - Biopesticide effect of green compost against fusarium wilt on melon plants. AB - AIMS: The biopesticide effect of four green composts against fusarium wilt in melon plants and the effect of soil quality in soils amended with composts were assayed. METHODS AND RESULTS: The composts consisted of pruning wastes, with or without addition of coffee wastes (3/1 and 4/1, dry wt/dry wt) or urea (1000/1, dry wt/dry wt). In vitro experiments suggested the biopesticide effect of the composts against Fusarium oxysporum, while only the compost of pine bark and urea (1000/1dry wt/dry wt) had an abiotic effect. Melon plant growth with composts and F. oxysporum was one to four times greater than in the non-amended soil, although there was no significant decrease in the level of the F. oxysporum in the soil. The addition of composts to the soil also improved its biological quality, as assessed by microbiological and biochemical parameters: ATP and hydrolases involved in the P (phosphatase), C (beta-glucosidase) and N (urease) cycles. CONCLUSIONS: Green composts had greater beneficial characteristics, improved plant growth and controlled fusarium wilt in melon plants. These composts improve the soil quality of semi-arid agricultural soils. SIGNIFICANCE AND IMPACT OF THE STUDY: Biotic and abiotic factors from composts have been tested as responsible of their biopesticide activity against fusarium wilt. PMID- 15752331 TI - A beta-glucosidase gene isolated from wine Lactobacillus plantarum is regulated by abiotic stresses. AB - AIMS: Little genetic information exists on the ability of wine lactic acid bacteria (LAB) to hydrolyse glycoconjugates during malolactic fermentation. We tried to fill this important gap by characterizing a gene codifying for a putative beta-glucosidase enzyme from wine Lactobacillus plantarum and from a commercial strain of Oenococcus oeni. METHODS AND RESULTS: The coding region of the putative beta-glucosidase gene is 1400 nucleotides long and started with an ATG codon. The gene is widespread among LAB and the highest identity was observed between the nucleotide of L. plantarum, Lactobacillus pentosus, Lactobacillus paraplantarum and O. oenibeta-glucosidase gene. The protein sequence deduced from the isolated genes has a calculated molecular mass of 61.19 kDa. Furthermore, the expression of the beta-glucosidase gene in L. plantarum strain was analysed, under several stress, by reverse transcriptase (RT)-PCR and Northern-blot analysis. The gene was apparently regulated by abiotic stresses such as temperature, ethanol and pH. CONCLUSIONS: The beta-glucosidase gene is widespread among LAB and its expression is probably regulated by a wide range of abiotic stresses. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibitory effect of temperature and ethanol on the L. plantarumbeta-glucosidase gene may be useful to explain the differences found in beta-glucosidase activity reported in wines by several authors. PMID- 15752332 TI - The effect of sulphur dioxide and oxygen on the viability and culturability of a strain of Acetobacter pasteurianus and a strain of Brettanomyces bruxellensis isolated from wine. AB - AIMS: The objective of this study was to investigate the effects of free molecular and bound forms of sulphur dioxide and oxygen on the viability and culturability of a selected strain of Acetobacter pasteurianus and a selected strain of Brettanomyces bruxellensis in wine. METHODS AND RESULTS: Acetic acid bacteria and Brettanomyces/Dekkera yeasts associated with wine spoilage were isolated from bottled commercial red wines. One bacterium, A. pasteurianus strain A8, and one yeast, B. bruxellensis strain B3a, were selected for further study. The resistance to sulphur dioxide and the effect of oxygen addition on these two selected strains were determined by using plating and epifluorescence techniques for monitoring cell viability in wine. Acetobacter pasteurianus A8 was more resistant to sulphur dioxide than B. bruxellensis B3a, with the latter being rapidly affected by a short exposure time to free molecular form of sulphur dioxide. As expected, neither of these microbial strains was affected by the bound form of sulphur dioxide. The addition of oxygen negated the difference observed between plate and epifluorescence counts for A. pasteurianus A8 during storage, while it stimulated growth of B. bruxellensis B3a. CONCLUSIONS: Acetobacter pasteurianus A8 can survive under anaerobic conditions in wine in the presence of sulphur dioxide. Brettanomyces bruxellensis B3a is more sensitive to sulphur dioxide than A. pasteurianus A8, but can grow in the presence of oxygen. Care should be taken to exclude oxygen from contact with wine when it is being transferred or moved. SIGNIFICANCE AND IMPACT OF THE STUDY: Wine spoilage can be avoided by preventing growth of undesirable acetic acid bacteria and Brettanomyces/Dekkera yeasts through the effective use of sulphur dioxide and the management of oxygen throughout the winemaking process. PMID- 15752333 TI - Chitosan inactivates spoilage yeasts but enhances survival of Escherichia coli O157:H7 in apple juice. AB - AIMS: To develop new measures for controlling both spoilage and pathogenic micro organisms in unpasteurized apple juice using chitosan. METHODS AND RESULTS: Micro organisms were isolated and identified from apple juice treated or untreated with chitosan using enrichment, selective media, microscopy, substrate assimilation patterns and ribosomal DNA profiling. Chitosan (0.05-0.1%) delayed spoilage by yeasts at 25 degrees C for up to 12 days but the effect was species specific: Kloeckera apiculata and Metschnikowia pulcherrima were inactivated but Saccharomyces cerevisiae and Pichia spp. multiplied slowly. In challenge experiments at 25 degrees C, total yeast counts were 3-5 log CFU ml(-1) lower in chitosan-treated juices than in the controls for 4 days but the survival of Escherichia coli O157:H7 was extended from 1 to 2 days; at 4 degrees C, chitosan reduced the yeast counts by 2-3 log CFU ml(-1) for up to 10 days but survival of the pathogen was prolonged from 3 to 5 days. The survival of Salmonella enterica serovar Typhimurium was unaffected by chitosan at either temperature. CONCLUSIONS: The addition of chitosan to apple juice delayed spoilage by yeasts but enhanced the survival of E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that the use of chitosan in the treatment of fruit juices may potentially lead to an increased risk of food poisoning from E. coli O157:H7. PMID- 15752334 TI - Purification, amino acid sequence and characterization of Bacthuricin F4, a new bacteriocin produced by Bacillus thuringiensis. AB - AIMS: Purification and characterization of a new bacteriocin, Bacthuricin F4 of Bacillus thuringiensis. METHODS AND RESULTS: A newly isolated B. thuringiensis subsp. kurstaki strain BUPM4, was shown to produce a novel bacteriocin named Bacthuricin F4. The highest bacteriocin activity was found in the growth medium and evidenced in the late exponential growth phase. Bacthuricin F4 could be purified by a two-step procedure: ammonium sulphate precipitation of protein from culture supernatant followed by a reverse phase chromatography. Upon purification, the specific activity was increased 100-fold. This bacteriocin was heat-stable up to 70 degrees C and resisted up to pH 3.0. Bacthuricin F4 was sensitive to proteases demonstrating its proteinaceous nature. Its molecular mass, determined by mass spectrometry was 3160.05 Da. Direct N-terminal sequencing of Bacthuricin F4 revealed the following sequence: DWTXWSXL. The latter was unique in the databases. Bacthuricin F4 was active against Bacillus species while it had little or no effect on Gram-negative bacteria. CONCLUSIONS: A strain BUPM4 of B. thuringiensis subsp. kurstaki, was shown to produce a new bacteriocin named Bacthuricin F4 of both new molecular mass (3160.05 Da) and new amino acid terminal sequence. This is, to our knowledge, the first bacteriocin exhibiting such characteristics reported to be produced by B. thuringiensis. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacteriocin produced by the B. thuringiensis strain BUPM4 respond to both criteria of thermostability and stability to low pHs. Thus, it could be used for the control of the related species of Bacillus harmful for agricultural products. PMID- 15752335 TI - Detection of Helicobacter pylori DNA in human faeces and water with different levels of faecal pollution in the north-east of Spain. AB - AIMS: To assess the role of water in the faecal transmission of Helicobacter pylori by detecting the DNA of this pathogen in human faecal samples and environmental water samples with a range of faecal pollution from the north-east of Spain. METHODS AND RESULTS: Semi-nested PCR was used to detect H. pylori in stools and water, both matrices with a complex biota. DNA was detected using highly specific primers of an ureA gene fragment. In addition, antigens were used to detect the bacteria in stools. Helicobacter pylori was detected in 33% of 36 human faecal samples and in 66% of wastewater samples, and 11% of river samples, but in none of the spring waters samples. Faecal pollution of the aquatic environment was tested analysing the presence of microbial indicators. CONCLUSIONS: We report the presence of H. pylori DNA in stools and in aquatic environments with different levels of faecal pollution, from the north-east of Spain. In this study a higher number of positive results were obtained in the more faecally polluted waters. SIGNIFICANCE AND IMPACT OF THE STUDY: These data indicate that water may be a vector of H. pylori in its faecal-oral route. PMID- 15752336 TI - Excision vs sponge swabbing - a comparison of methods for the microbiological sampling of beef, pork and lamb carcasses. AB - AIMS: The aim of this research was to compare excision sampling with polyurethane and cellulose acetate sponge swabbing for the recovery of total viable counts and Enterobacteriaceae on meat carcasses. METHODS AND RESULTS: Two sample types were used to compare the methods: (i) individual samples, taken from four sites on each carcass and (ii) composite samples, created by pooling the samples from four sites from an additional set of carcasses. When the polyurethane sponge and excision method were compared for individual sites, there were no significant differences in bacterial recovery on beef and pork carcasses and on two of four sites on lamb carcasses. However, when samples from each site were pooled, the excision method was more efficient than either swabbing method across the three animal species. CONCLUSIONS: Sampling using the polyurethane sponge represents an equivalent alternative method to excision for the bacteriological sampling of carcass surfaces which is nondestructive and less labour intensive. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides the scientific basis for using sponge swabbing instead of excision in compliance with 2001/471/EC. PMID- 15752337 TI - Response of the microbial community to copper oxychloride in acidic sandy loam soil. AB - AIMS: Determining the response of different microbial parameters to copper oxychloride in acidic sandy loam soil samples using cultivation-dependent and direct microscopic techniques. METHODS AND RESULTS: Culturable microbial populations were monitored for 245 days in a series of soil microcosms spiked with different copper oxychloride concentrations. Microbial populations responded differently to additional Cu. Protistan numbers and soil metabolic potential decreased. Experiments with more soil samples revealed that metabolic potential was not significantly affected by < or =100 mg kg(-1) additional Cu. However, a negative impact on protista was noted in soil containing only 15 mg kg(-1) EDTA extractable Cu. The negative impact on protistan numbers was less severe in soils with a higher phosphorous and zinc content. CONCLUSIONS: Bacterial populations responded differently, and protista were most sensitive to elevated Cu levels. Protistan numbers in soil from uncultivated land were higher and seemed to be more sensitive to additional Cu than the numbers of these organisms in soil originating from cultivated land. SIGNIFICANCE AND IMPACT OF THE STUDY: Protistan sensitivity to small increases in Cu levels demonstrates the vulnerability of the soil ecosystem to Cu perturbations, especially when the importance of protista as link in the flow of energy between trophic levels is considered. PMID- 15752338 TI - Evidence of septic system failure determined by a bacterial biochemical fingerprinting method. AB - AIMS: To provide evidence of septic system failure by comparing two faecal indicator bacteria, enterococci and Escherichia coli, from defective septic tanks and adjacent creeks. METHODS AND RESULTS: A biochemical fingerprinting method was used to type and compare enterococci and E. coli strains from 39 septic tanks with creek water samples. Phenotypic diversity of enterococci (0.5 +/- 0.3) and E. coli (0.5 +/- 0.3) in septic tanks were significantly lower than those found in water samples (0.8 +/- 0.1, P < 0.0001 for enterococci and 0.9 +/- 0.1, P < 0.0001 for E. coli). Among 1072 enterococci isolates tested from septic tanks, 203 biochemical phenotypes (BPTs) were found of which 98 BPTs from 33 septic tanks were identical to several water samples. Similarly, among 621 E. coli isolates tested from septic tanks, 159 BPTs were found of which 53 BPTs from 26 septic tanks were also identical to water samples. The number of the latter bacteria was significantly (P = 0.01) higher in water samples collected from downstream compared with that of upstream in the study area. A high similarity between the populations of both indicator bacteria was also found between defective septic tanks and downstream water samples further indicating the contamination of both creeks by defective septic systems. CONCLUSIONS: Biochemical fingerprinting of faecal indicator bacteria is a useful and rapid method to provide direct evidence for septic system failure. Combination of both faecal indicator bacteria (enterococci and E. coli) provides a better judgement of the performance of a septic system. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to provide direct evidence of septic system failure by identifying the presence of specific bacterial types in septic tanks and surface waters. Based on our findings, we suggest that the performance evaluation of a septic system should be accompanied by direct analysis of faecal indicator bacteria. PMID- 15752339 TI - Dissemination of transferable CTX-M-type extended-spectrum beta-lactamase producing Escherichia coli in Korea. AB - AIMS: Among 365 Escherichia coli isolated in 2003, 31 cefotaxime-resistant isolates were obtained from clinical specimens taken from adults hospitalized in Busan, Korea. Six extended-spectrum beta-lactamase (ESBL)-producing isolates were investigated further to determine the mechanism of resistance. METHODS AND RESULTS: These isolates were analysed by antibiotic susceptibility testing, pI determination, plasmid profiles, transconjugation test, PCR-restriction fragment length polymorphism (RFLP), enterobacterial repetitive consensus (ERIC)-PCR and DNA sequencing. All six of these isolates were found to contain the CTX-M-type ESBL genes. Five clinical isolates and their transconjugants produced CTX-M-3. One clinical isolate (K17391) and its transconjugant (trcK17391) produced CTX-M 15. Five clinical isolates also produced another TEM-1. One clinical isolate (K12776) also contained another TEM-52. CTX-M-3 ESBL gene was responsible for the resistance to piperacillin, cephalothin, cefotaxime, cefepime and aztreonam. CTX M-15 or TEM-52 was especially responsible for the resistance to ceftazidime. CONCLUSIONS: These results appear to represent the in vivo evolution of CTX-M type beta-lactamase genes (bla(CTX-M-3) --> bla(CTX-M-15)) under the selective pressure of antimicrobial therapy (especially ceftazidime). PCR-RFLP is a reliable method to discriminate CTX-M-15 gene from CTX-M-3 gene. ERIC-PCR analysis revealed that dissemination of CTX-M-3 was not due to a clonal outbreak of a resistant strain but to the intra-species spread of resistance to piperacillin, cephalothin, cefotaxime, cefepime and aztreonam in Korea. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the occurrence of CTX-M-1 cluster ESBLs in Korea. A more comprehensive survey of these ESBL types from Korea is urgently needed because of the in vivo evolution of CTX-M-15 from CTX-M-3. The emergence of these CTX-M-type ESBLs suggests that diagnostic laboratories should screen for ESBLs with ceftazidime as well as cefotaxime; they should still perform clavulanate synergy tests on resistant isolates. PMID- 15752340 TI - Highly diverse variable number tandem repeat loci in the E. coli O157:H7 and O55:H7 genomes for high-resolution molecular typing. AB - AIM: Evaluation of the Escherichia coli genome for variable number tandem repeat (VNTR) loci in order to provide a subtyping tool with greater discrimination and more efficient capacity. METHODS AND RESULTS: Twenty-nine putative VNTR loci were identified from the E. coli genomic sequence. Their variability was validated by characterizing the number of repeats at each locus in a set of 56 E. coli O157:H7/HN and O55:H7 isolates. An optimized multiplex assay system was developed to facility high capacity analysis. Locus diversity values ranged from 0.23 to 0.95 while the number of alleles ranged from two to 29. This multiple-locus VNTR analysis (MLVA) data was used to describe genetic relationships among these isolates and was compared with PFGE (pulse field gel electrophoresis) data from a subset of the same strains. Genetic similarity values were highly correlated between the two approaches, through MLVA was capable of discrimination amongst closely related isolates when PFGE similar values were equal to 1.0. CONCLUSIONS: Highly variable VNTR loci exist in the E. coli O157:H7 genome and are excellent estimators of genetic relationships, in particular for closely related isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli O157:H7 MLVA offers a complimentary analysis to the more traditional PFGE approach. Application of MLVA to an outbreak cluster could generate superior molecular epidemiology and result in a more effective public health response. PMID- 15752341 TI - Effects of a biocontrol agent and methyl jasmonate on postharvest diseases of peach fruit and the possible mechanisms involved. AB - AIMS: To investigate effects of application of 200 micromol l(-1) methyl jasmonate [MeJA (200)] and Cryptococcus laurentii alone or in combination against postharvest diseases (Monilinia fructicola and Penicillium expansum) in peach fruit stored at 25 and 0 degrees C, and to evaluate the possible mechanisms involved. METHODS AND RESULTS: The efficacy of controlling postharvest diseases by resistance induced in peach fruit treated with MeJA (200) and C. laurentii alone or in combination and the relationship between activities of defence related enzymes in peach fruit and lesions caused by M. fructicola and P. expansum were examined. At the same time, the effects of MeJA (200) on the population of C. laurentii in the peach wounds and on the mycelial growth of M. fructicola and P. expansumin vitro were investigated. The results indicated that treatment of peach fruit with C. laurentii at 1 x 10(8) CFU ml(-1) alone, or combining C. laurentii at 5 x 10(7) CFU ml(-1) with MeJA (200) all resulted in a lower lesion diameter of brown rot and blue mould caused by M. fructicola and P. expansum compared with the controls in peach fruit. MeJA (200) enhanced the population of C. laurentii, and inhibited mycelial growth of P. expansum. However, it had a little effect on M. fructicolain vitro. MeJA and C. laurentii alone or in combination induced higher activities of Chitinase, beta-1,3 glucanase, phenylalanine ammonia-lyase and peroxidase (POD) than applying the yeast alone at both 25 and 0 degrees C. CONCLUSIONS: MeJA (200) not only directly inhibited mycelial spread of postharvest pathogens, but also increased population of C. laurentii, which induced stronger disease resistance in fruit than MeJA or yeast alone, and resulted in a lower lesion diameter of brown rot and blue mould caused by M. fructicola and P. expansum. SIGNIFICANCE AND IMPACT OF THE STUDY: MeJA (200) in combination with C. laurentii was beneficial for controlling brown rot and blue mould caused by M. fructicola and P. expansum in peach fruit. The inhibitory mechanism was mainly because of resistance induced in peach fruit by MeJA and C. laurentii. In addition, direct inhibition of MeJA on P. expansum also played a role in controlling blue mould. PMID- 15752342 TI - mRNA detection by reverse transcription-PCR for monitoring viability and potential virulence in a pathogenic strain of Vibrio parahaemolyticus in viable but nonculturable state. AB - AIMS: This work investigates the maintenance of viability and potential virulence of Vibrio parahaemolyticus in a viable but nonculturable population (VBNC) state by reverse transcription-polymerase chain reaction (RT-PCR). METHODS AND RESULTS: Housekeeping genes, 16S-23S rDNA and rpoS, as well as virulence genes, tdh1 and tdh2, were selected and detected by PCR in a pathogenic strain of V. parahaemolyticus (Vp4). Their expression was then studied by RT-PCR in V. parahaemolyticus Vp4 cultivated in rich medium at 37 degrees C. The 16S-23S rDNA and rpoS, tdh1, tdh2 genes were transcripted at the mid-logarithmic, stationary and late stationary phases, corresponding to various physiological states. The expression of these genes was also studied by RT-PCR in a VBNC population of V. parahaemolyticus Vp4 in artificial seawater (ASW). The effect of temperature (washing of bacterial culture and microcosms) on the attaining VBNC bacteria was first considered. Washing of V. parahaemolyticus Vp4, collected at the mid logarithmic phase, at 10 or 4 degrees C before inoculation in ASW at 4 degrees C allowed bacteria entered the VBNC state between 22 and 31 days. The 16S-23S rDNA and rpoS gene were expressed in the VBNC bacteria whereas no expression of the tdh1 and tdh2 genes was observed in the same populations. CONCLUSION: The two selected housekeeping genes, 16S-23S rDNA and rpoS, proved to be good viability markers for V. parahaemolyticus Vp4 in culturable and VBNC states. These first data indicated that the pathogenic strain Vp4 would not maintain the expression of the virulence genes, tdh1 and tdh2, in VBNC state. SIGNIFICANCE AND IMPACT OF THE STUDY: Use of RT-PCR for investigating the maintenance or not of viability and potential virulence in VBNC V. parahaemolyticus will facilitate further study to evaluate the potential risk presented by this pathogen in the environment. PMID- 15752343 TI - Improving the specificity of 16S rDNA-based polymerase chain reaction for detecting Borrelia burgdorferi sensu lato-causative agents of human Lyme disease. AB - AIMS: 16S rDNA sequences of Borrelia burgdorferi sensu lato were aligned with the 16S rDNA sequences of Borrelia hermsii, Borrelia turicatae, and Borrelia lonestari in order to identify primers that might be used to more specifically identify agents of human Lyme disease in ticks in human skin samples. METHODS AND RESULTS: Standard polymerase chain reaction (PCR), using an oligonucleotide sequence, designated TEC1, was shown, in combination with a previously developed primer (LD2) to amplify strains of B. burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii, but not the non-Lyme causing B. hermsii or B. turicatae. This primer pair, designated Bbsl, was successfully used to amplify B. burgdorferi sensu lato from skin biopsies of patients with Lyme disease symptoms as well as from Ixodes scapularis, Amblyomma americanum and Dermacentor variabilis ticks. CONCLUSIONS: The primer set Bbsl allows for the rapid detection and differentiation of B. burgdorferi sensu lato from non-Lyme disease-causing Borrelia species in ticks and human tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR primer set, Bbsl, will greatly facilitate detection of the causative agents of Lyme disease in infected ticks and human skin samples assisting in epidemiological studies, and potentially allowing for a more rapid diagnosis of the disease in patients. PMID- 15752344 TI - Combined application of methods to taxonomic identification of Saccharomyces strains in fermenting botrytized grape must. AB - AIMS: Although numerous physiological and molecular methods have been proposed for yeast taxonomy, the unambiguous separation of Saccharomyces sensu stricto species in natural samples is still an incompletely resolved issue. In this study the power of various methods was compared in the identification of strains isolated from fermenting botrytized grape musts. METHODS AND RESULTS: Conventional taxonomic and physiological tests and molecular methods developed for rapid identification were used. CONCLUSIONS: None of the methods tested was sufficiently powerful. However, the combination of electrophoretic karyotyping and the PCR-RFLP of MET2 with growth tests at 10 and 37 degrees C provided results sufficient for species identification of Saccharomyces wine strains which were not interspecific hybrids or recombinants. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed combination of molecular and physiological methods allows specific taxonomic identification and separation of Saccharomyces wine strains without extensive genetic and molecular analysis. The proposed combined approach can also identify hybrids and recombinants. PMID- 15752345 TI - The occurrence of Campylobacter subtypes in environmental reservoirs and potential transmission routes. AB - AIM: To identify potential reservoirs and transmission routes of human pathogenic Campylobacter spp. METHODS AND RESULTS: An enrichment PCR method for the detection and identification of Campylobacter jejuni and/or Campylobacter coli in faecal, food and river water samples was applied to 1450 samples of 12 matrix types obtained from a defined geographical area. PCR-positive samples were cultured to yield isolates for typing, and the data for 616 C. jejuni isolates obtained. Serotyping and SmaI macrorestriction profiling using pulsed field gel electrophoresis revealed a high level of diversity within the isolates from each matrix. Campylobacter jejuni and C. coli subtypes indistinguishable from those obtained from human cases were detected in most of the matrices examined. No Campylobacter isolates were isolated from possum faeces. CONCLUSIONS: Ten of the 12 matrices examined may be involved in the transmission of human campylobacteriosis as they contained Campylobacter subtypes also isolated from clinical cases. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicate that, for this rural population, a range of potential transmission routes that could lead to campylobacteriosis exist. Their relative importance needs to be assessed from an exposure assessment standpoint. PMID- 15752346 TI - Berry phenolics selectively inhibit the growth of intestinal pathogens. AB - AIMS: To investigate the effects of berries and berry phenolics on pathogenic intestinal bacteria and to identify single phenolic compounds being responsible for antimicrobial activity. METHODS AND RESULTS: Antimicrobial activity of eight Nordic berries and their phenolic extracts and purified phenolic fractions were measured against eight selected human pathogens. Pathogenic bacterial strains, both Gram-positive and Gram-negative, were selectively inhibited by bioactive berry compounds. Cloudberry and raspberry were the best inhibitors, and Staphylococcus and Salmonella the most sensitive bacteria. Phenolic compounds, especially ellagitannins, were strong inhibitory compounds against Staphylococcus bacteria. Salmonella bacteria were only partly inhibited by the berry phenolics, and most of the inhibition seemed to originate from other compounds, such as organic acids. Listeria strains were not affected by berry compounds, with the exception of cranberry. Phenolic compounds affect the bacteria in different mechanisms. CONCLUSIONS: Berries and their phenolics selectively inhibit the growth of human pathogenic bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Antimicrobial properties of berries could be utilized in functional foods. Furthermore these compounds would be of high interest for further evaluation of their properties as natural antimicrobial agents for food and pharmaceutical industry. PMID- 15752347 TI - Detection of lactococcal 936-species bacteriophages in whey by magnetic capture hybridization PCR targeting a variable region of receptor-binding protein genes. AB - AIMS: To develop PCR assays able to distinguish between groups within lactococcal 936-species bacteriophages, as defined by their different receptor-binding protein (RBP) genes. METHODS AND RESULTS: DNA sequences of RBP genes from 17 lactococcal bacteriophages of the 936-species were compared, and six phage groups were identified. For each phage group a specific primer pair targeting a variable region of the RBP genes was designed. In nine of 20 whey samples, from dairies with recorded phage problems, between one and six phage groups were identified by conventional PCR. The sensitivity and specificity of the method was improved by magnetic capture hybridization (MCH)-PCR using a capture probe targeting an 80-bp highly conserved region just upstream from the RBP gene in all the investigated phages. The MCH-PCR was performed on 100 microl whey samples and the detection limit of the assay was 10(2)-10(3) PFU ml(-1) as opposed to the detection limit of 10(4) PFU ml(-1) for conventional PCR performed on 1-microl whey samples. CONCLUSIONS: In this study, PCR assays have been developed to detect six different types of RBP genes in lactococcal 936-species bacteriophages. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR assays have practical applications at cheese plants for detection of 936-species phages with different RBP and thereby potentially with different host ranges. This knowledge will make it possible to improve starter culture rotation systems in the dairy industry. PMID- 15752349 TI - Oxidative stress in the hippocampus after pilocarpine-induced status epilepticus in Wistar rats. AB - The role of oxidative stress in pilocarpine-induced status epilepticus was investigated by measuring lipid peroxidation level, nitrite content, GSH concentration, and superoxide dismutase and catalase activities in the hippocampus of Wistar rats. The control group was subcutaneously injected with 0.9% saline. The experimental group received pilocarpine (400 mg.kg(-1), subcutaneous). Both groups were killed 24 h after treatment. After the induction of status epilepticus, there were significant increases (77% and 51%, respectively) in lipid peroxidation and nitrite concentration, but a 55% decrease in GSH content. Catalase activity was augmented 88%, but superoxide dismutase activity remained unaltered. These results show evidence of neuronal damage in the hippocampus due to a decrease in GSH concentration and an increase in lipid peroxidation and nitrite content. GSH and catalase activity are involved in mechanisms responsible for eliminating oxygen free radicals during the establishment of status epilepticus in the hippocampus. In contrast, no correlations between superoxide dismutase and catalase activities were observed. Our results suggest that GSH and catalase activity play an antioxidant role in the hippocampus during status epilepticus. PMID- 15752350 TI - Increased NADPH concentration obtained by metabolic engineering of the pentose phosphate pathway in Aspergillus niger. AB - Many biosynthetic reactions and bioconversions are limited by low availability of NADPH. With the purpose of increasing the NADPH concentration and/or the flux through the pentose phosphate pathway in Aspergillus niger, the genes encoding glucose 6-phosphate dehydrogenase (gsdA), 6-phosphogluconate dehydrogenase (gndA) and transketolase (tktA) were cloned and overexpressed in separate strains. Intracellular NADPH concentration was increased two- to ninefold as a result of 13-fold overproduction of 6-phosphogluconate dehydrogenase. Although overproduction of glucose 6-phosphate dehydrogenase and transketolase changed the concentration of several metabolites it did not result in increased NADPH concentration. To establish the effects of overexpression of the three genes, wild-type and overexpressing strains were characterized in detail in exponential and stationary phase of bioreactor cultures containing minimal media, with glucose as the carbon source and ammonium or nitrate as the nitrogen source and final cell density limiting substrate. Enzymes, intermediary metabolites, polyol pools (intra- and extracellular), organic acids, growth rates and rate constant of induction of acid production in postexponential phase were measured. None of the modified strains had a changed growth rate. Partial least square regressions showed the correlations between NADPH and up to 40 other variables (concentration of enzymes and metabolites) and it was possible to predict the intracellular NADPH concentration from relatively easily obtainable data (the concentration of enzymes, polyols and oxalate). This prediction might be used in screening for high NADPH levels in engineered strains or mutants of other organisms. PMID- 15752351 TI - Effect of coenzyme modification on the structural and catalytic properties of wild-type transketolase and of the variant E418A from Saccharomyces cerevisiae. AB - Transketolase from baker's yeast is a thiamin diphosphate-dependent enzyme in sugar metabolism that reconstitutes with various analogues of the coenzyme. The methylated analogues (4'-methylamino-thiamin diphosphate and N1'-methylated thiamin diphosphate) of the native cofactor were used to investigate the function of the aminopyrimidine moiety of the coenzyme in transketolase catalysis. For the wild-type transketolase complex with the 4'-methylamino analogue, no electron density was found for the methyl group in the X-ray structure, whereas in the complex with the N1'-methylated coenzyme the entire aminopyrimidine ring was disordered. This indicates a high flexibility of the respective parts of the enzyme-bound thiamin diphosphate analogues. In the E418A variant of transketolase reconstituted with N1'-methylated thiamin diphosphate, the electron density of the analogue was well defined and showed the typical V-conformation found in the wild-type holoenzyme [Lindqvist Y, Schneider G, Ermler U, Sundstrom M (1992) EMBO J11, 2373-2379]. The near-UV CD spectrum of the variant E418A reconstituted with N1'-methylated thiamin diphosphate was identical to that of the wild-type holoenzyme, while the CD spectrum of the variant combined with the unmodified cofactor did not overlap with that of the native protein. The activation of the analogues was measured by the H/D-exchange at C2. Methylation at the N1' position of the cofactor activated the enzyme-bound cofactor analogue (as shown by a fast H/D-exchange rate constant). The absorbance changes in the course of substrate turnover of the different complexes investigated (transient kinetics) revealed the stability of the alpha-carbanion/enamine as the key intermediate in cofactor action to be dependent on the functionality of the 4-aminopyrimidine moiety of thiamin diphosphate. PMID- 15752352 TI - Dehydroepiandrosterone inhibits the proliferation of human umbilical vein endothelial cells by enhancing the expression of p53 and p21, restricting the phosphorylation of retinoblastoma protein, and is androgen- and estrogen-receptor independent. AB - Dehydroepiandrosterone (DHEA), a steroid hormone, modified the proliferation of human umbilical vein endothelial cells in a dose-dependent manner. Its inactive sulfate ester (DHEA-S) and two of its metabolites -- estradiol and testosterone - had no inhibitory effect at physiological concentrations. Antiproliferation was associated with arrest in the G1 phase of the cell cycle, but not with cell death, as evaluated by cleavage of poly(ADP-ribose) polymerase and exposure of phosphatidylserine. The effect was not blocked by inhibitors of androgen or estrogen receptors. DHEA diminished the levels of phosphorylated retinoblastoma protein and increased the expression of p53 and p21 mRNAs. These results show that DHEA inhibits endothelial cell proliferation by regulating cell cycle relevant proteins through a cytoplasmic steroid hormone-independent pathway. PMID- 15752353 TI - Entamoeba histolytica TATA-box binding protein binds to different TATA variants in vitro. AB - The ability of Entamoeba histolytica TATA binding protein (EhTBP) to interact with different TATA boxes in gene promoters may be one of the key factors to perform an efficient transcription in this human parasite. In this paper we used several TATA variants to study the in vitro EhTBP DNA-binding activity and to determine the TATA-EhTBP dissociation constants. The presence of EhTBP in complexes formed by nuclear extracts (NE) and the TATTTAAA oligonucleotide, which corresponds to the canonical TATA box for E. histolytica, was demonstrated by gel shift assays. In these experiments a single NE-TATTTAAA oligonucleotide complex was detected. Complex was retarded by anti-EhTBP Igs in supershift experiments and antibodies also recognized the cross-linked complex in Western blot assays. Recombinant EhTBP formed specific complexes with TATA variants found in E. histolytica gene promoters and other TATA variants generated by mutation of TATTTAAA sequence. The dissociation constants of recombinant EhTBP for TATA variants ranged between 1.04 (+/-0.39) x 10(-11) and 1.60 (+/-0.37) x 10(-10) m. TATTTAAA and TAT_ _AAA motifs presented the lowest KD values. Intriguingly, the recombinant EhTBP affinity for TATA variants is stronger than other TBPs reported. In addition, EhTBP is more promiscuous than human and yeast TBPs, probably due to modifications in amino acids involved in TBP-DNA binding. PMID- 15752354 TI - Conserved pore-forming regions in polypeptide-transporting proteins. AB - Transport of solutes and polypeptides across membranes is an essential process for every cell. In the past, much focus has been placed on helical transporters. Recently, the beta-barrel-shaped transporters have also attracted some attention. The members of this family are found in the outer bacterial membrane and the outer membrane of endosymbiotically derived organelles. Here we analyze the features and the evolutionary development of a specified translocator family, namely the beta-barrel-shaped polypeptide-transporters. We identified sequence motifs, which characterize all transporters of this family, as well as motifs specific for a certain subgroup of proteins of this class. The general motifs are related to the structural composition of the pores. Further analysis revealed a defined distance of two motifs to the C-terminal portion of the proteins. Furthermore, the evolutionary relationship of the proteins and the motifs are discussed. PMID- 15752355 TI - N-terminal deletion of the gamma subunit affects the stabilization and activity of chloroplast ATP synthase. AB - Five truncation mutants of chloroplast ATP synthase gamma subunit from spinach (Spinacia oleracea) lacking 8, 12, 16, 20 or 60 N-terminal amino acids were generated by PCR by a mutagenesis method. The recombinant gamma genes were overexpressed in Escherichia coli and assembled with alphabeta subunits into a native complex. The wild-type (WT) alphabetagamma assembly i.e. alphabetagammaWT exhibited high (Mg2+)-dependent and (Ca2+)-dependent ATP hydrolytic activity. Deletions of eight residues of the gamma subunit N-terminus caused a decrease in rates of ATP hydrolysis to 30% of that of the alphabetaWT assembly. Furthermore, only approximately 6% of ATP hydrolytic activity was retained with the sequential deletions of gamma subunit up to 20 residues compared with the activity of the alphabetaWT assembly. The inhibitory effect of the epsilon subunit on ATP hydrolysis of these alphabetagamma assemblies varied to a large extent. These observations indicate that the N-terminus of the gamma subunit is very important, together with other regions of the gamma subunit, in stabilization of the enzyme complex or during cooperative catalysis. In addition, the in vitro binding assay showed that the gamma subunit N-terminus is not a crucial region in binding of the epsilon subunit. PMID- 15752356 TI - Characterization of alpha-synuclein aggregation and synergistic toxicity with protein tau in yeast. AB - A yeast model was generated to study the mechanisms and phenotypical repercussions of expression of alpha-synuclein as well as the coexpression of protein tau. The data show that aggregation of alpha-synuclein is a nucleation elongation process initiated at the plasma membrane. Aggregation is consistently enhanced by dimethyl sulfoxide, which is known to increase the level of phospholipids and membranes in yeast cells. Aggregation of alpha-synuclein was also triggered by treatment of the yeast cells with ferrous ions, which are known to increase oxidative stress. In addition, data are presented in support of the hypothesis that degradation of alpha-synuclein occurs via autophagy and proteasomes and that aggregation of alpha-synuclein disturbs endocytosis. Reminiscent of observations in double-transgenic mice, coexpression of alpha synuclein and protein tau in yeast cells is synergistically toxic, as exemplified by inhibition of proliferation. Taken together, the data show that these yeast models recapitulate major aspects of alpha-synuclein aggregation and cytotoxicity, and offer great potential for defining the underlying mechanisms of toxicity and synergistic actions of alpha-synuclein and protein tau. PMID- 15752357 TI - Caspase-2 is resistant to inhibition by inhibitor of apoptosis proteins (IAPs) and can activate caspase-7. AB - Caspases are a family of cysteine proteases with roles in cytokine maturation or apoptosis. Caspase-2 was the first pro-apoptotic caspase identified, but its functions in apoptotic signal transduction are still being elucidated. This study examined the regulation of the activity of caspase-2 using recombinant proteins and a yeast-based system. Our data suggest that for human caspase-2 to be active its large and small subunits must be separated. For maximal activity its prodomain must also be removed. Consistent with its proposed identity as an upstream caspase, caspase-2 could provoke the activation of caspase-7. Caspase-2 was not subject to inhibition by members of the IAP family of apoptosis inhibitors. PMID- 15752358 TI - Flavonol 3-O-glycoside hydroxycinnamoyltransferases from Scots pine (Pinus sylvestris L.). AB - Flavonol 3-O-glucosides esterified with ferulic or p-coumaric acid at positions 3'' and 6'' are the major UV-B screening pigments of the epidermal layer of Scots pine (Pinus sylvestris) needles. The last steps in the biosynthesis of these compounds are catalyzed by enzymes that transfer the acyl part of hydroxycinnamic acid CoA esters to flavonol 3-O-glucosides. A newly developed enzyme assay revealed three flavonol 3-O-glucoside hydroxycinnamoyltransferases (HCTs) in Scots pine needles with specificities for positions 3'', 4'' or 6''. The positions of the acyl groups were identified by cochromatography with reference compounds and by NMR spectroscopy. The enzymes were characterized by molecular mass, isoelectric point, and also pH and temperature optima. Substrate specificities for flavonol glycosides and hydroxycinnamic acid CoA esters as well as kinetic properties of 3''- and 6''HCT suggested that acylation preferably occurs with glucosides and p-coumaroyl-CoA. In addition, acylation takes place in a well-defined order, beginning at position 6'' followed by acylation at position 3''. These results give the first detailed characterization of flavonol 3-O glycoside HCTs involved in the protection of plant tissues against UV-B (280-315 nm) radiation. PMID- 15752360 TI - Allosteric properties of the GTP activated and CTP inhibited uracil phosphoribosyltransferase from the thermoacidophilic archaeon Sulfolobus solfataricus. AB - The upp gene, encoding uracil phosphoribosyltransferase (UPRTase) from the thermoacidophilic archaeon Sulfolobus solfataricus, was cloned and expressed in Escherichia coli. The enzyme was purified to homogeneity. It behaved as a tetramer in solution and showed optimal activity at pH 5.5 when assayed at 60 degrees C. Enzyme activity was strongly stimulated by GTP and inhibited by CTP. GTP caused an approximately 20-fold increase in the turnover number kcat and raised the Km values for 5-phosphoribosyl-1-diphosphate (PRPP) and uracil by two- and >10-fold, respectively. The inhibition by CTP was complex as it depended on the presence of the reaction product UMP. Neither CTP nor UMP were strong inhibitors of the enzyme, but when present in combination their inhibition was extremely powerful. Ligand binding analyses showed that GTP and PRPP bind cooperatively to the enzyme and that the inhibitors CTP and UMP can be bound simultaneously (KD equal to 2 and 0.5 microm, respectively). The binding of each of the inhibitors was incompatible with binding of PRPP or GTP. The data indicate that UPRTase undergoes a transition from a weakly active or inactive T-state, favored by binding of UMP and CTP, to an active R-state, favored by binding of GTP and PRPP. PMID- 15752359 TI - Identification of the heparin-binding domains of the interferon-induced protein kinase, PKR. AB - PKR is an interferon-induced serine-threonine protein kinase that plays an important role in the mediation of the antiviral and antiproliferative actions of interferons. PKR is present at low basal levels in cells and its expression is induced at the transcriptional level by interferons. PKR's kinase activity stays latent until it binds to its activator. In the case of virally infected cells, double-stranded (ds) RNA serves as PKR's activator. The dsRNA binds to PKR via two copies of an evolutionarily conserved motif, thus inducing a conformational change, unmasking the ATP-binding site and leading to autophosphorylation of PKR. Activated PKR then phosphorylates the alpha-subunit of the protein synthesis initiation factor 2 (eIF2alpha) thereby inducing a general block in the initiation of protein synthesis. In addition to dsRNA, polyanionic agents such as heparin can also activate PKR. In contrast to dsRNA-induced activation of PKR, heparin-dependent PKR activation has so far remained uncharacterized. In order to understand the mechanism of heparin-induced PKR activation, we have mapped the heparin-binding domains of PKR. Our results indicate that PKR has two heparin binding domains that are nonoverlapping with its dsRNA-binding domains. Although both these domains can function independently of each other, they function cooperatively when present together. Point mutations created within these domains rendered PKR defective in heparin-binding, thereby confirming their essential role. In addition, these mutants were defective in kinase activity as determined by both in vitro and in vivo assays. PMID- 15752361 TI - Influence of divalent cations on the structural thermostability and thermal inactivation kinetics of class II xylose isomerases. AB - The effects of divalent metal cations on structural thermostability and the inactivation kinetics of homologous class II d-xylose isomerases (XI; EC 5.3.1.5) from mesophilic (Escherichia coli and Bacillus licheniformis), thermophilic (Thermoanaerobacterium thermosulfurigenes), and hyperthermophilic (Thermotoga neapolitana) bacteria were examined. Unlike the three less thermophilic XIs that were substantially structurally stabilized in the presence of Co2+ or Mn2+ (and Mg2+ to a lesser extent), the melting temperature [(Tm) approximately 100 degrees C] of T. neapolitana XI (TNXI) varied little in the presence or absence of a single type of metal. In the presence of any two of these metals, TNXI exhibited a second melting transition between 110 degrees C and 114 degrees C. TNXI kinetic inactivation, which was non-first order, could be modeled as a two-step sequential process. TNXI inactivation in the presence of 5 mm metal at 99-100 degrees C was slowest in the presence of Mn2+[half-life (t(1/2)) of 84 min], compared to Co2+ (t(1/2) of 14 min) and Mg2+ (t(1/2) of 2 min). While adding Co2+ to Mg2+ increased TNXI's t(1/2) at 99-100 degrees C from 2 to 7.5 min, TNXI showed no significant activity at temperatures above the first melting transition. The results reported here suggest that, unlike the other class II XIs examined, single metals are required for TNXI activity, but are not essential for its structural thermostability. The structural form corresponding to the second melting transition of TNXI in the presence of two metals is not known, but likely results from cooperative interactions between dissimilar metals in the two metal binding sites. PMID- 15752362 TI - Functional analysis of the methylmalonyl-CoA epimerase from Caenorhabditis elegans. AB - Methylmalonyl-CoA epimerase (MCE) is an enzyme involved in the propionyl-CoA metabolism that is responsible for the degradation of branched amino acids and odd-chain fatty acids. This pathway typically functions in the reversible conversion of propionyl-CoA to succinyl-CoA. The Caenorhabditis elegans genome contains a single gene encoding MCE (mce-1) corresponding to a 15 kDa protein. This was expressed in Escherichia coli and the enzymatic activity was determined. Analysis of the protein expression pattern at both the tissue and subcellular level by microinjection of green fluorescent protein constructs revealed expression in the pharynx, hypodermis and, most prominently in body wall muscles. The subcellular pattern agrees with predictions of mitochondrial localization. The sequence similarity to an MCE of known structure was high enough to permit a three-dimensional model to be built, suggesting conservation of ligand and metal binding sites. Comparison with corresponding sequences from a variety of organisms shows more than 1/6 of the sequence is completely conserved. Mutants allelic to mce-1 showed no obvious phenotypic alterations, demonstrating that the enzyme is not essential for normal worm development under laboratory conditions. However, survival of the knockout mutants was altered when exposed to stress conditions, with mutants surprisingly showing an increased resistance to oxidative stress. PMID- 15752364 TI - Autonomous oscillations in Saccharomyces cerevisiae during batch cultures on trehalose. AB - We report that autonomous oscillations, which usually happen in aerobic glucose limited continuous cultures of yeast at low dilution rate, were also observed in trehalose discontinuous cultures of Saccharomyces cerevisiae. This unexpected oscillatory behaviour was therefore examined using fast Fourier transformation of online gas measurements. This robust mathematical analysis underlined the existence of two types of oscillation. The first was found to be linked to the cell cycle because (a) the periodicity corresponded to a fraction of the generation time and (b) the oscillations were accompanied by a transient increase in the budding index, mobilization of storage carbohydrates, and fermentative activity. Moreover, these oscillations occurred in a range of specific growth rates between 0.04 and 0.15 h(-1). All these criteria were consistent with the cell-cycle-related metabolic oscillations observed in the same range of growth rates in glucose-limited continuous cultures. The second type were short-period respiratory oscillations, independent of the specific growth rate. Both types of oscillation were found to take place consecutively and/or simultaneously during batch culture on trehalose. In addition, mobilization of intracellular trehalose emerged as a key parameter for the sustainability of these autonomous oscillations as they were no longer observed in a mutant defective in neutral trehalase activity. We propose that batch culture on trehalose may be an excellent device for further investigation of the molecular mechanisms that underlie autonomous oscillations in yeast. PMID- 15752363 TI - A novel glycogen-targeting subunit of protein phosphatase 1 that is regulated by insulin and shows differential tissue distribution in humans and rodents. AB - Stimulation of glycogen-targeted protein phosphatase 1 (PP1) activity by insulin contributes to the dephosphorylation and activation of hepatic glycogen synthase (GS) leading to an increase in glycogen synthesis. The glycogen-targeting subunits of PP1, GL and R5/PTG, are downregulated in the livers of diabetic rodents and restored by insulin treatment. We show here that the mammalian gene PPP1R3E encodes a novel glycogen-targeting subunit of PP1 that is expressed in rodent liver. The phosphatase activity associated with R3E is slightly higher than that associated with R5/PTG and it is downregulated in streptozotocin induced diabetes by 60-70% and restored by insulin treatment. Surprisingly, although mRNA for R3E is most highly expressed in rat liver and heart muscle, with only low levels in skeletal muscle, R3E mRNA is most abundant in human skeletal muscle and heart tissues with barely detectable levels in human liver. This species-specific difference in R3E mRNA expression has similarities to the high level of expression of GL mRNA in human but not rodent skeletal muscle. The observations imply that the mechanisms by which insulin regulates glycogen synthesis in liver and skeletal muscle are different in rodents and humans. PMID- 15752365 TI - Identification of alternative promoter usage for the matrix Gla protein gene. Evidence for differential expression during early development in Xenopus laevis. AB - Recent cloning of the Xenopus laevis (Xl) matrix Gla protein (MGP) gene indicated the presence of a conserved overall structure for this gene between mammals and amphibians but identified an additional 5'-exon, not detected in mammals, flanked by a functional, calcium-sensitive promoter, 3042 bp distant from the ATG initiation codon. DNA sequence analysis identified a second TATA-like DNA motif located at the 3' end of intron 1 and adjacent to the ATG-containing second exon. This putative proximal promoter was found to direct transcription of the luciferase reporter gene in the X. laevis A6 cell line, a result confirmed by subsequent deletion mutant analysis. RT-PCR analysis of XlMGP gene expression during early development identified a different temporal expression of the two transcripts, strongly suggesting differential promoter activation under the control of either maternally inherited or developmentally induced regulatory factors. Our results provide further evidence of the usefulness of nonmammalian model systems to elucidate the complex regulation of MGP gene transcription and raise the possibility that a similar mechanism of regulation may also exist in mammals. PMID- 15752366 TI - Structural characterization of Ca2+/CaM in complex with the phosphorylase kinase PhK5 peptide. AB - Phosphorylase kinase (PhK) is a large hexadecameric enzyme consisting of four copies of four subunits: (alphabetagammadelta)4. An intrinsic calmodulin (CaM, the delta subunit) binds directly to the gamma protein kinase chain. The interaction site of CaM on gamma has been localized to a C-terminal extension of the kinase domain. Two 25-mer peptides derived from this region, PhK5 and PhK13, were identified previously as potential CaM-binding sites. Complex formation between Ca2+/CaM with these two peptides was characterized using analytical gel filtration and NMR methods. NMR chemical shift perturbation studies showed that while PhK5 forms a robust complex with Ca2+/CaM, no interactions with PhK13 were observed. 15N relaxation characteristics of Ca2+/CaM and Ca2+/CaM/PhK5 complexes were compared with the experimentally determined structures of several Ca2+/CaM/peptide complexes. Good fits were observed between Ca2+/CaM/PhK5 and three structures: Ca2+/CaM complexes with peptides from endothelial nitric oxide synthase, with smooth muscle myosin light chain kinase and CaM kinase I. We conclude that the PhK5 site is likely to have a direct role in Ca2+-regulated control of PhK activity through the formation of a classical 'compact' CaM complex. PMID- 15752367 TI - An engineered disulfide bridge mimics the effect of calcium to protect neutral protease against local unfolding. AB - The extreme thermal stabilization achieved by the introduction of a disulfide bond (G8C/N60C) into the cysteine-free wild-type-like mutant (pWT) of the neutral protease from Bacillus stearothermophilus[Mansfeld J, Vriend G, Dijkstra BW, Veltman OR, Van den Burg B, Venema G, Ulbrich-Hofmann R & Eijsink VG (1997) J Biol Chem272, 11152-11156] was attributed to the fixation of the loop region 56 69. In this study, the role of calcium ions in the guanidine hydrochloride (GdnHCl)-induced unfolding and autoproteolysis kinetics of pWT and G8C/N60C was analyzed by fluorescence spectroscopy, far-UV CD spectroscopy and SDS/PAGE. First order rate constants (kobs) were evaluated by chevron plots (ln kobs vs. GdnHCl concentration). The kobs of unfolding showed a difference of nearly six orders of magnitude (DeltaDeltaG# = 33.5 kJ.mol(-1) at 25 degrees C) between calcium saturation (at 100 mM CaCl2) and complete removal of calcium ions (in the presence of 100 mM EDTA). Analysis of the protease variant W55F indicated that calcium binding-site III, situated in the critical region 56-69, determines the stability at calcium ion concentrations between 5 and 50 mM. In the chevron plots the disulfide bridge in G8C/N60C shows a similar effect compared with pWT as the addition of calcium ions, suggesting that the introduced disulfide bridge fixes the region (near calcium binding-site III) that is responsible for unfolding and subsequent autoproteolysis. Owing to the presence of the disulfide bridge, the DeltaDeltaG# is 13.2 kJ.mol(-1) at 25 degrees C and 5 mM CaCl2. Non-linear chevron plots reveal an intermediate in unfolding probably caused by local unfolding of the loop 56-69. The occurrence of this intermediate is prevented by calcium concentrations of > 5 mM, or the introduction of the disulfide bridge G8C/N60C. PMID- 15752368 TI - Inhibitory properties of cystatin F and its localization in U937 promonocyte cells. AB - Cystatin F is a recently discovered type II cystatin expressed almost exclusively in immune cells. It is present intracellularly in lysosome-like vesicles, which suggests a potential role in regulating papain-like cathepsins involved in antigen presentation. Therefore, interactions of cystatin F with several of its potential targets, cathepsins F, K, V, S, H, X and C, were studied in vitro. Cystatin F tightly inhibited cathepsins F, K and V with Ki values ranging from 0.17 nM to 0.35 nM, whereas cathepsins S and H were inhibited with 100-fold lower affinities (Ki approximately 30 nM). The exopeptidases, cathepsins C and X were not inhibited by cystatin F. In order to investigate the biological significance of the inhibition data, the intracellular localization of cystatin F and its potential targets, cathepsins B, H, L, S, C and K, were studied by confocal microscopy in U937 promonocyte cells. Although vesicular staining was observed for all the enzymes, only cathepsins H and X were found to be colocalized with the inhibitor. This suggests that cystatin F in U937 cells may function as a regulatory inhibitor of proteolytic activity of cathepsin H or, more likely, as a protection against cathepsins misdirected to specific cystatin F containing endosomal/lysosomal vesicles. The finding that cystatin F was not colocalized with cystatin C suggests distinct functions for these two cysteine protease inhibitors in U937 cells. PMID- 15752369 TI - Liver X receptor agonists inhibit tissue factor expression in macrophages. AB - Exposure of blood to tissue factor (TF) rapidly initiates the coagulation serine protease cascades. TF is expressed by macrophages and other types of cell within atherosclerotic lesions and plays an important role in thrombus formation after plaque rupture. Macrophage TF expression is induced by pro-inflammatory stimuli including lipopolysaccharide (LPS), interleukin-1beta and tumor necrosis factor alpha. Here we demonstrate that activation of liver X receptors (LXRs) LXRalpha and LXRbeta suppresses TF expression. Treatment of mouse peritoneal macrophages with synthetic LXR agonist T0901317 or GW3965 reduced TF expression induced by pro-inflammatory stimuli. LXR agonists also suppressed TF expression and its activity in human monocytes. Human and mouse TF promoters contain binding sites for the transcription factors AP-1, NFkappaB, Egr-1 and Sp1, but no LXR-binding sites could be found. Cotransfection assays with LXR and TF promoter constructs in RAW 264.7 cells revealed that LXR agonists suppressed LPS-induced TF promoter activity. Analysis of TF promoter also showed that inhibition of TF promoter activity by LXR was at least in part through inhibition of the NFkappaB signaling pathway. In addition, in vivo, LXR agonists reduced TF expression within aortic lesions in an atherosclerosis mouse model as well as in kidney and lung in mice stimulated with LPS. These findings indicate that activation of LXR results in reduction of TF expression, which may influence atherothrombosis in patients with vascular disease. PMID- 15752370 TI - Moving about. PMID- 15752371 TI - Comparative effects of glyceryl trinitrate and amyl nitrite on pulse wave reflection and augmentation index. AB - AIMS: The influence of vasodilators on augmentation index (AIx) offers a simple, rapid and noninvasive method of evaluating vascular function. Glyceryl trinitrate (GTN) is widely used as an endothelium-independent vasodilator, although other nitrates that are shorter acting may have advantages in clinical studies. The aim of this study was to compare the effects of two short-acting nitrates, GTN and amyl nitrite, which have differing pharmacodynamic profiles. METHODS: Twenty-one healthy volunteers (15 male; mean age 35 years, range 21-56 years) attended on three occasions and received sublingual GTN (0.5 mg for 3 min), inhaled amyl nitrite (0.2 ml inhaled for 30 s), or no treatment in a randomized cross-over design. Haemodynamic responses of AIx, blood pressure and thoracic bioimpedance (heart rate, cardiac index) were assessed by measurement at baseline, every 60 s for the first 5 min, and then every 5 min for a further 55 min. RESULTS: AIx was reduced by amyl nitrite (peak effect -9 +/- 2% at 1 min, P < 0.002) and GTN (peak effect -12 +/- 3% at 4 min, P < 0.05). Compared with amyl nitrite, the onset and offset of action of GTN was slower. Amyl nitrite initially increased heart rate by 27 +/- 4% (P < 0.001) and cardiac index by 13 +/- 3% (P < 0.001) whereas GTN had no significant effect (P > 0.05). Neither agent affected blood pressure. CONCLUSIONS: GTN causes a slower and more sustained reduction in AIx than amyl nitrite. Although amyl nitrite causes a more rapid fall and recovery in AIx, it induces a reflex tachycardia that may limit interpretation of the initial (1 min) but not later (2 min) changes in AIx. The prolonged offset of GTN suggests that a sufficient washout period must be included when making repeated measures or when assessing the subsequent effects of other agents. PMID- 15752372 TI - Mycophenolic acid pharmacokinetics and related outcomes early after renal transplant. AB - AIMS: The pharmacokinetics of mycophenolic acid and its glucuronide are complex. This study investigated the pharmacokinetics, pharmacodynamics and protein binding of mycophenolic acid and its glucuronide metabolite, early post transplant in renal allograft recipients. METHODS: Forty-two de novo renal transplant recipients receiving mycophenolate mofetil and concomitant cyclosporin (n = 32) or tacrolimus (n = 10) participated in the study. Blood samples were taken on day 5 post-transplant for measurement of free and total concentrations of mycophenolic acid, mycophenolic acid glucuronide and relevant biochemistry. Associations between free fraction and biochemistry were investigated. Free and total 6-h area under the concentration-time curve (AUC0-6) of mycophenolic acid was assessed relative to clinical outcomes in the first month post-transplant. RESULTS: Kinetic variability of free and total mycophenolic acid and its glucuronide was greater in patients on cyclosporin (12- to 18-fold variation) than on tacrolimus (four- to fivefold) cotherapy. Cyclosporin-treated patients also had significantly lower predose total mycophenolic acid concentrations than tacrolimus-treated patients (median 0.8 mg l(-1) and 1.6 mg l(-1), respectively, P = 0.002). Mycophenolic acid glucuronide predose concentration correlated positively with mycophenolic acid glucuronide AUC0-6 (r > 0.95). Mycophenolic acid free fraction varied 11-fold, from 1.6% to 18.3%, whilst the glucuronide free fraction varied threefold, from 17.4% to 54.1%. Urea and creatinine concentrations correlated positively (r > 0.46), whilst albumin correlated negatively (r = -0.54) with free fraction of mycophenolic acid. Similar relationships were found for the free fraction of mycophenolic acid glucuronide. Mycophenolic acid free fraction was on average 70% higher in patients with albumin concentrations below a specified albumin cut-off concentration of 31 g l( 1)[free fraction = 7 +/- 4% for lower albumin and 4 +/- 3% for higher albumin, respectively; P = 0.001; 95% confidence interval (CI) for the difference 1.9, 4.2]. Neither free nor total mycophenolic acid AUC0-6 was related to rejection (P > 0.07). Free AUC0-6 was significantly higher in those patients with thrombocytopenic, leukopenic and/or infectious outcomes than in those without (mean +/- SD 1.9 +/- 0.3 mg h(-1) l(-1) and 1.1 +/- 0.1 mg h(-1) l(-1), P = 0.0043; 95% CI for the difference 0.3, 1.4). CONCLUSIONS: The marked variability in mycophenolic acid/glucuronide pharmacokinetics occurring early post-transplant during the current study was greater in cyclosporin (12-18-fold) than in tacrolimus (four- to fivefold) treated patients. Concomitant cyclosporin was associated with total mycophenolic acid concentrations approximately half that of tacrolimus. Patients with marked renal impairment had the highest free fractions reported to date. The exposure to unbound mycophenolic acid was significantly related to infections and haematological toxicity. PMID- 15752373 TI - Dosing strategy for enoxaparin in patients with renal impairment presenting with acute coronary syndromes. AB - BACKGROUND: Phase III clinical studies have confirmed that enoxaparin is superior to standard heparin in reducing the rate of recurrent ischaemic events in patients with non-ST elevation acute coronary syndromes. Patients with moderate to severe renal impairment were, however, excluded from these studies. Due to the hydrophilic disposition of enoxaparin, accumulation is likely in patients with renal dysfunction, thereby increasing the risk of haemorrhagic complications if standard weight adjusted treatment doses are used. Arbitrary dose reduction has been reported to increase the risk of ischaemic events, presumably due to inadequate enoxaparin concentrations. AIM: The aims of this study were to investigate the influence of glomerular filtration rate (GFR) on the pharmacokinetics of subcutaneously administered enoxaparin, and to develop a practical dosing algorithm in renal impairment that can easily be used at the bedside. METHODS: Thirty-eight patients, median age 78 years (range 44-87), mean GFR 32 ml min(-1) (range 16-117) and mean weight 69 kg (range 32-95), presenting with acute coronary syndrome were recruited into the study. Approximately 10 anti Xa concentrations were taken per patient over their period of therapy. A population pharmacokinetic model was developed using non linear mixed effects modelling techniques, utilizing the software NONMEM. Stochastic simulations were performed to identify the most suitable dosing regimen. RESULTS: Three hundred and thirteen anti-Xa concentrations were collected. A two compartment, first order input model was identified as the best baseline model. Covariates found to improve model fitting were GFR as a linear function on clearance (CL) and weight as a linear function on the central volume compartment (Vc). The fraction of drug excreted unchanged (Fu) was estimated at 71%. CL and Vc from the final covariate model were estimated as; CL (l h(-1)) = 0.681 per 4.8 l hr(-1) (GFR) + 0.229 Vc (l) = 5.22 per 80 kg (total body weight) CONCLUSIONS: Clearance of enoxaparin was predictably related to GFR estimated using the Cockroft and Gault equation, with ideal body weight used as the size descriptor. According to our model no dosage adjustment from the standard 1.0 mg kg(-1) 12 hourly is required for the first 48 h of treatment. Maintenance doses thereafter can be calculated using standard proportional adjustments based on Fu equal to 0.71. PMID- 15752374 TI - Pharmacokinetics of pitavastatin in subjects with Child-Pugh A and B cirrhosis. AB - AIM: Lipid lowering therapy with 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors is increasingly used for the prevention of cardiovascular events, but they should be used with caution in patients with impaired liver function. We therefore studied the pharmacokinetics of pitavastatin in patients with liver cirrhosis. METHODS: Plasma concentrations of pitavastatin were determined after administration of 2 mg single-dose pitavastatin to 12 male patients with liver cirrhosis (six Child-Pugh grade A and six grade B). These results were compared with the single-dose pharmacokinetic results obtained from six male volunteers without liver disease. RESULTS: Administration of 2 mg single dose pitavastatin to patients with Child-Pugh grade A and grade B cirrhosis resulted in a 1.19- and 2.47-fold increase in Cmax and 1.27- and 3.64-fold increase in AUCt, respectively, when compared with normal subjects. The geomean Cmax of pitavastatin was 59.5 ng ml(-1), 70.7 ng ml(-1) and 147.1 ng ml(-1) in the control, Child-Pugh grade A and Child-Pugh grade B groups, respectively. The geomean AUCt of pitavastatin in the three groups was 121.2 ng h(-1) ml(-1), 154.2 ng h(-1) ml(-1) and 441.7 ng h(-1) ml(-1), respectively. The geomean Cmax of pitavastatin lactone was 20.3 ng ml(-1), 19.1 ng ml(-1) and 9.9 ng ml(-1) in the control, Child-Pugh grade A and grade B groups, respectively. The AUCt of pitavastatin lactone was 120.2 h(-1) ml(-1), 108.8 h(-1) ml(-1) and 87.5 h(-1) ml(-1), respectively. CONCLUSION: The plasma concentration of pitavastatin is increased in patients with liver cirrhosis. In such patients, caution is required, although dose reduction may not be necessary in Child-Pugh A cirrhosis. PMID- 15752375 TI - The disposition of oral amodiaquine in Papua New Guinean children with falciparum malaria. AB - AIMS: We assessed the disposition of oral amodiaquine (AQ) and CYP2C8 polymorphism in 20 children with falciparum malaria. METHODS: AQ and DEAQ concentrations were determined with SPE-HPLC method. CYP2C8 genotypes were assessed by PCR-RFLP method. RESULTS: AQ was not detectable beyond day 3 postdose. Cmax for DEAQ was reached in 3.0 days. The mean values for t1/2, MRT, and AUCtotal were 10.1 days, 15.5 days and 4512.6 microg l(-1) day, respectively. All the children were CYP2C8* homozygous. CONCLUSION: Our data are consistent with those previously reported, and the AQ regimen seems pharmacokinetically adequate in the absence of CYP2C8 polymorphism. PMID- 15752376 TI - Effects of clarithromycin on lansoprazole pharmacokinetics between CYP2C19 genotypes. AB - AIMS: Lansoprazole is a substrate of CYP2C19 and CYP3A. The aim of this study was to compare the inhibitory effects of clarithromycin, an inhibitor of CYP3A on the metabolism of lansoprazole between CYP2C19 genotypes. METHODS: A two-way randomized double-blind, placebo-controlled crossover study was performed. Eighteen volunteers, of whom six were homozygous extensive metabolizers (EMs), six were heterozygous EMs and six were poor metabolizers (PMs) for CYP2C19, received two 6-day courses of either clarithromycin 800 mg or placebo daily in a randomized fashion with a single oral dose of lansoprazole 60 mg on day 6 in all cases. Plasma concentrations of lansoprazole and its metabolites, 5 hydroxylansoprazole and lansoprazole sulphone were monitored up to 24 h after dosing. RESULTS: During placebo administration, the mean AUC0, infinity of lansoprazole in homozygous EMs, heterozygous EMs and PMs were 4652 (95% CI, 2294, 7009) ng ml(-1) h, 8299 (4784, 11814) ng ml(-1) h and 25293 (17643, 32943) ng ml( 1) h (P < 0.001), respectively. Clarithromycin treatment significantly increased Cmax by 1.47-fold, 1.71-fold and 1.52-fold and AUC0, infinity of lansoprazole by 1.55-fold, 1.74-fold, and 1.80-fold in these genotype groups, respectively, whereas elimination half-life was prolonged only in PMs. The clarithromycin mediated percent increase in pharmacokinetic parameters such as Cmax, AUC0, infinity or elimination half-life did not differ between the three CYP2C19 genotypes. CONCLUSIONS: The present study indicates that there are significant drug interactions between lansoprazole and clarithromycin in all CYP2C19 genotype groups probably through CYP3A inhibition. The bioavailability of lansoprazole might, to some extent, be increased through inhibition of P-glycoprotein during clarithromycin treatment. PMID- 15752377 TI - Association between CYP1A2 activity and riluzole clearance in patients with amyotrophic lateral sclerosis. AB - AIMS: Riluzole is used in a fixed dosing schedule of 50 mg twice daily to treat patients with amyotropic lateral sclerosis (ALS), one form of motor neurone disease. The large variability in the pharmacokinetics of riluzole may be a factor contributing to its limited therapeutic benefit. Riluzole is assumed to be mainly metabolized by the cytochrome P450 enzyme 1A2 (CYP1A2). The aim of the study was to investigate the relationship between CYP1A2 activity and riluzole clearance with a view to optimize drug treatment. METHODS: A group of 30 ALS patients participated in the study. In each patient the CYP1A2 activity was determined using caffeine as a metabolic probe. Riluzole clearance was estimated from serum drug concentration measurements followed by Bayesian fitting. RESULTS: Riluzole clearance and the serum paraxanthine : caffeine (P/C) ratio showed a positive correlation (r = 0.693; P = 0.0002). Linear regression analysis identified the P/C ratio (beta: 1.16) and height (beta: 0.027) as independent predictors of riluzole clearance (adjusted r2 = 0.369). CONCLUSIONS: The P/C ratio, used as measure of CYP1A2 activity, significantly correlated with the riluzole clearance, although only 37% of the observed variability could be explained. PMID- 15752378 TI - Melphalan pharmacokinetics in children with malignant disease: influence of body weight, renal function, carboplatin therapy and total body irradiation. AB - AIM: To analyse the pharmacokinetics of melphalan in 52 children (0.3-18 years) and determine whether any clinical factors affect the pharmacokinetic parameters Additionally, to examine whether a test melphalan dose can predict the pharmacokinetics of a full dose, when there are 5 intervening days of carboplatin therapy. METHODS: Melphalan concentrations were measured in 14 blood samples collected from each child following doses ranging from 30 to 180 mg m(-2). The pharmacokinetics were analysed with Kinetica 4.0. RESULTS: Children who did not have carboplatin (n = 27) had median melphalan clearance (CL) of 15.5 l h(-1) m( 2) (interquartile range: 12.4-19.9 l h(-1) m(-2)) and steady state volume of distribution (Vss) of 14.9 l m(-2) (interquartile range: 12.7-18.3 l m(-2)). Children who had carboplatin (n = 25) had 34% lower median CL (10.2 l h(-1) m( 2)) and 18% lower median Vss (12.2 l m(-2)) (P < 0.001). Melphalan elimination was impaired in a separate group of three children given concomitant carboplatin and etoposide. Stepwise multiple linear regression indicated that weight, carboplatin, glomerular filtration rate (GFR) and total body irradiation (TBI) significantly affected CL, while weight and carboplatin influenced Vss. A test dose (10 mg m(-2)) tended to underpredict the area-under-the-concentration-vs. time-curve for a full (180 mg m(-2)) dose in 19 individuals given carboplatin. CONCLUSIONS: In children, melphalan CL is influenced by weight, carboplatin, TBI and GFR. Vss is influenced by weight and carboplatin. PMID- 15752379 TI - Population pharmacokinetics of orally administered paclitaxel formulated in Cremophor EL. AB - AIM: The vehicle Cremophor EL (CrEL) has been shown to impair the absorption of paclitaxel by micellar entrapment of the drug in the gastrointestinal tract. The goal of this study was to develop a semimechanistic population pharmacokinetic model to study the influence of CrEL on the oral absorption of paclitaxel. METHOD: Paclitaxel plasma-concentration time profiles were available from 55 patients (M:F, 17 : 38; total 67 courses; 797 samples), receiving paclitaxel orally once or twice daily (dose range 60-360 mg m(-2)) together with 12-15 mg kg(-1) cyclosporin A. A population pharmacokinetic model was developed using the nonlinear mixed effect modelling program NONMEM. RESULTS: After absorption, paclitaxel pharmacokinetics were best described using a two-compartment model with linear distribution from the central compartment into a peripheral compartment and first-order elimination. Paclitaxel in the gastrointestinal tract was modelled as free fraction or bound to CrEL, with only the free fraction available for absorption into the central compartment. The equilibrium between free and bound paclitaxel was influenced by the concentration of CrEL present in the gastrointestinal tract. The concentration of CrEL in the gastrointestinal tract decreased with time with a first order rate constant of 1.73 h(-1). The bioavailability of paclitaxel was independent of the dose and of CrEL. Estimated apparent paclitaxel clearance and volume of distribution were 127 l h(-1) and 409 l, respectively. Large interpatient variability was observed. Covariate analysis did not reveal significant relationships with any of the pharmacokinetic parameters. CONCLUSION: A pharmacokinetic model was developed that described the pharmacokinetics of orally administered paclitaxel. CrEL strongly influenced paclitaxel absorption from the gastrointestinal tract resulting in time-dependent but no significant dose-dependent absorption over the examined dose range studied. PMID- 15752380 TI - Mechanistic pharmacokinetic modelling of ephedrine, norephedrine and caffeine in healthy subjects. AB - AIM: The combination of ephedrine and caffeine has been used in herbal products for weight loss and athletic performance-enhancement, but the pharmacokinetic profiles of these compounds have not been well characterized. This study aimed to develop a mechanistic model describing ephedrine, norephedrine, and caffeine pharmacokinetics and their interactions in healthy subjects. METHODS: The pharmacokinetic model was developed based on the simultaneous modelling using plasma samples gathered from two clinical trials. The treatments consisted of single-doses of pharmaceutical caffeine and ephedrine, given alone or together, and an herbal formulation containing both caffeine and ephedrine. We used a mixed effect statistical model and the program NONMEM to take account of intersubject variability. RESULTS: Three hundred and seventy-nine ephedrine, 352 norephedrine, 417 caffeine plasma concentrations and 40 ephedrine urine concentrations were obtained from 24 subjects. A one-compartment model with first-order absorption described the caffeine data. Caffeine clearance was 0.083 l min(-1) (CV 38%) and decreased to 0.038 l min(-1) in presence of oral contraceptive therapy, its volume of distribution was 38.6 l (CV 20%) and its absorption rate constant was 0.064 l min(-1) (CV 50%). A four-compartment model described the pharmocokinetics of ephedrine and norephedrine. Ephedrine was eliminated mostly renally, with a clearance of 0.34 l min(-1) (CV 11%), and a volume of distribution of 181 l (CV 19%). Nonlinearity in the conversion of ephedrine to norephedrine was observed. Different models showed that the simultaneous administration of caffeine, or the amount of caffeine in the absorption compartment, was associated with a slower rate of absorption of ephedrine. A 32% greater relative bioavailability of herbal compared with pharmaceutical ephedrine administration was observed. CONCLUSIONS: We describe a mechanistic model for ephedrine, norephedrine and caffeine pharmacokinetics and their interactions. The relative bioavailability of ephedrine differed between the herbal supplement compared with the pharmaceutical formulation. Concomitant ingestion of caffeine slowed the absorption rate of ephedrine, which is mainly related to the amount of the former in the absorption compartment. A saturable process appears to be involved in the metabolism of ephedrine to norephedrine. PMID- 15752381 TI - Co-administration of ketoconazole with H1-antagonists ebastine and loratadine in healthy subjects: pharmacokinetic and pharmacodynamic effects. AB - AIMS: Two studies were conducted to evaluate the effects of coadministration of ketoconazole with two nonsedating antihistamines, ebastine and loratadine, on the QTc interval and on the pharmacokinetics of the antihistamines. METHODS: In both studies healthy male subjects (55 in one study and 62 in the other) were assigned to receive 5 days of antihistamine (ebastine 20 mg qd in one study, and loratadine 10 mg qd in the other) or placebo alone using a predetermined randomization schedule, followed by 8 days of concomitant ketoconazole 450 mg qd/antihistamine or ketoconazole 400 mg qd/placebo. Serial ECGs and blood sampling for drug analysis were performed at baseline and on study days 5 (at the end of monotherapy) and 13 (at the end of combination therapy). QT intervals were corrected for heart rate using the formula QTc = QT/RR(alpha) with special emphasis on individualized alpha values derived from each subject's own QT/RR relationship at baseline. RESULTS: No significant changes in QTc interval from baseline were observed after 5 days administration of ebastine, loratadine or placebo. Ketoconazole/placebo increased the mean QTc (95% CI) by 6.96 (3.31 10.62) ms in the ebastine study and by 7.52 (4.15-10.89) ms in the loratadine study. Mean QTc was statistically significantly increased during both ebastine/ketoconazole administration (12.21 ms; 7.39-17.03 ms) and loratadine/ketoconazole administration (10.68 ms; 6.15-15.21 ms) but these changes were not statistically significantly different from the increases seen with placebo/ketoconazole (6.96 ms; 3.31-10.62 ms), P = 0.08 ebastine study, (7.52 ms; 4.15-10.89 ms), P = 0.26 loratadine study). After the addition of ketoconazole, the mean area under the plasma concentration-time curve (AUC) for ebastine increased by 42.5 fold, and that of its metabolite carebastine by 1.4 fold. The mean AUC for loratadine increased by 4.5 fold and that of its metabolite desloratadine by 1.9 fold following administration of ketoconazole. No subjects were withdrawn because of ECG changes or drug-related adverse events. CONCLUSIONS: Ketoconazole altered the pharmacokinetic profiles of both ebastine and loratadine although the effect was greater for the former drug. The coadministration of ebastine with ketoconazole resulted in a non significant mean increase of 5.25 ms (-0.65 to 11.15 ms) over ketoconazole with placebo (6.96 ms) while ketoconazole plus loratadine resulted in a nonsignificant mean increase of 3.16 ms (-2.73 to 8.68 ms) over ketoconazole plus placebo (7.52 ms). Changes in uncorrected QT intervals for both antihistamines were not statistically different from those observed with ketoconazole alone. The greater effect of ketoconazole on the pharmacokinetics of ebastine was not accompanied by a correspondingly greater pharmacodynamic effect on cardiac repolarization. PMID- 15752382 TI - A predictive model for exemestane pharmacokinetics/pharmacodynamics incorporating the effect of food and formulation. AB - AIMS: Exemestane (Aromasin) is an irreversible aromatase inactivator used for the treatment of postmenopausal women with advanced breast cancer. The objective of this study was to evaluate the effect of formulation comparing a sugar-coated tablet (SCT) with a suspension and food on the pharmacokinetics (PK) and pharmacodynamics (PD) with respect to plasma estrone sulphate (E1S) concentrations of exemestane, using a PK/PD approach. METHODS: This was an open, three-period, randomized, crossover study. Twelve healthy postmenopausal women received single oral doses of 25 mg exemestane as a SCT after fasting or food and as a suspension after fasting. Exemestane and E1S concentrations were determined before and up to 14 days after drug administration. Population analysis was performed in two steps: (i) a compartmental PK model was selected incorporating the effect of food and formulation; (ii) conditional on the PK model, a PD model was developed employing indirect response models. Model selection was performed using standard statistical tests. Validation and assessment of the predictive capability of the selected model was performed using real test data sets obtained from the literature. RESULTS: A three-compartment model with first-order elimination rate best described exemestane disposition (k12 0.454, k21 0.158, k13 0.174, k31 0.016 and k 0.738 h(-1)). Absorption was described by a mono exponential function [ka 2.3 (SCT after fasting), 1.1 (SCT after food) and 7.6 h( 1) (suspension); lag time 0.2 h]. The PD model assumed that E1S plasma concentrations are determined by a zero-order synthesis rate (6.5 pg ml(-1) h( 1)) and a first-order elimination constant (0.032 h(-1)). Exemestane inhibited E1S synthesis with a C50 value of 22.1 pg ml(-1). The mean population estimates were used to simulate the administration of different doses of the drug (0.5, 1, 2.5, 5 and 25 mg day(-1)). The model predictions were in agreement with historical data. CONCLUSIONS: Exemestane absorption is influenced by the formulation of the drug and by food, but its disposition is independent of both. PK differences do no translate into clinically important differences in the PD. The PK/PD model developed was able to predict successfully the response to different doses and administration schedules with respect to oestrogen suppression. PMID- 15752383 TI - Relationship between the C3435T and G2677T(A) polymorphisms in the ABCB1 gene and P-glycoprotein expression in human liver. AB - AIMS: The aim of the study was to determine whether a correlation exists between MDR1 (ABCB1) gene polymorphisms at positions 3435 (C3435T) and 2677 (G2677T(A)) and the expression of human hepatic P-glycoprotein (P-gp). METHODS: P-gp protein expression in 26 human livers was assessed by Western blotting and ABCB1 mRNA expression was determined by real time RT-PCR. The C3435T and G2677T(A) polymorphisms were identified by RFLP and direct sequence analysis, respectively. RESULTS: The C and G allele frequencies for the C3435T and G2677T(A) polymorphisms were 0.48 and 0.79, respectively, and the genotypes were in Hardy Weinberg equilibrium. There was a 200- and 20-fold variation in the expression of ABCB1 mRNA and Pgp protein expression, respectively. There were no differences in mRNA and protein expression identified amongst the different genotypes attributable to the C3435T and G2677T(A) polymorphisms in the ABCB1 gene. Exposure to a PXR ligand prior to death did not influence mRNA or protein expression. CONCLUSIONS: There is substantial variability in the expression of Pgp in human liver, but this is not due to the presence of C3435T and G2677T(A) polymorphisms in the ABCB1 gene, although our study is limited by a small sample size. PMID- 15752387 TI - From crystal ball towards cognitive anaesthesiology. PMID- 15752384 TI - Paracetamol: a haemorrhagic risk factor in patients on warfarin. AB - AIM: To quantify the effect of paracetamol on the anticoagulant effect of warfarin under normal clinical conditions. PATIENTS AND METHODS: In a prospective double-blind, cross-over, placebo-controlled study, 11 patients on stable warfarin therapy received in random order two 14-day regimens of paracetamol 4 g day(-1) or placebo, with a 14-day or more wash-out period in between, time necessary to fulfil the inclusion criteria. RESULTS: In patients on paracetamol, the mean maximum increase in the International Normalized Ratio (INR) observed was 1.04 +/- 0.55 vs. 0.20 +/- 0.32 in those on placebo (P = 0.003). The mean maximum INR observed was significantly higher with paracetamol than with placebo (3.47 vs. 2.61, P = 0.01). In patients receiving paracetamol, the mean observed INR was significantly increased after 4 days (+ 0.6 +/- 0.6, P < 0.001). CONCLUSION: Paracetamol at 4 g day(-1) induces a significant increase in INR in patients receiving a stable regimen of warfarin, increasing the risk of bleeding associated with warfarin. PMID- 15752388 TI - Impact of the AEP-Monitor/2-derived composite auditory-evoked potential index on propofol consumption and emergence times during total intravenous anaesthesia with propofol and remifentanil in children. AB - BACKGROUND: The composite auditory evoked potential index (cAAI), derived from the AEP Monitor/2 (version 1.6; Danmeter A/S, Odense, Denmark) is a measure of the hypnotic component of general anaesthesia. The purpose of this study was to evaluate the impact of cAAI guidance on propofol consumption and emergence times in children receiving total intravenous anaesthesia (TIVA) with propofol and remifentanil. METHODS: Twenty children, aged 3-11 years, scheduled for strabismus repair under TIVA with propofol and remifentanil were enrolled. Remifentanil was given to all patients at a constant infusion rate of 0.3 microg kg(-1) min(-1) throughout the anaesthesia. Patients were randomly allocated to receive a continuous propofol infusion adjusted either according to a conventional clinical practice (Group C, n = 10) or guided by cAAI-monitoring (Group G, n = 10, target cAAI 25-35). All patients were connected to the AEP Monitor/2, but in group C the anaesthetist was blinded to cAAI values. Propofol consumption (mgkg(-1)h(-1)) and emergence times (min) were the primary and secondary outcome measures. RESULTS: Propofol consumption and emergence times (mean +/- SD) were significantly lower in group G compared to group C (Propofol: G: 4.2 +/- 1.7 vs. C 6.4 +/- 1.3 mg kg( 1) h(-1); P < 0.01; emergence times: G: 5.1 +/- 3.7 vs. C 13.2 +/- 8.2 min; P < 0.01). Intraoperative cAAI values (median [interquartile range]) were significantly higher in group G (23.9 [18-29.7]) than in group C (18.4 [16.0 22.1]; P < 0.01). Haemodynamic variables remained stable within age-related limits, and there were no observations of adverse events, especially no clinical signs of intraoperative awareness in any patient. CONCLUSION: Composite auditory evoked potential index monitoring during propofol/remifentanil-TIVA in children results in reduced propofol consumption and faster emergence. PMID- 15752389 TI - EEG spectral entropy, heart rate, photoplethysmography and motor responses to skin incision during sevoflurane anaesthesia. AB - BACKGROUND: Analgesia is a part of balanced anaesthesia, but direct indicators of nociception do not exist. We examined the relationship between motor reactions and physiological variables during skin incision in sevoflurane anaesthesia and hypothesized that nociception could be detected and graded by significant changes in these variables. METHODS: Thirty-one women scheduled for abdominal hysterectomy participated in the study. Anaesthesia was induced with fentanyl (1 microg kg(-1)), propofol (1 mg kg(-1)) and sevoflurane. Skin incision was performed 14 min after induction during 1.6% end-tidal sevoflurane anaesthesia without neuromuscular blockade. Electrocardiography (ECG), photoplethysmography (PPG) and electroencephalography (EEG) were registered, and a range of variables was computed from these signals. The postincision values, normalized with respect to their preincision values, of movers vs. non-movers were compared. The variables showing significant differences between movers and non-movers were used to develop a logistic regression equation for the classification of patients into movers or non-movers. RESULTS: Twenty-six patients were eligible for analysis, and 12 (46%) displayed a motor reaction to skin incision (movers). Many ECG, PPG and EEG-related variables showed significant differences between the pre- and postincision periods. The best classification performance, assessed by leave-one out cross-validation, between movers and non-movers was achieved with the combination of response entropy of EEG, RR-interval and PPG notch amplitude. The corresponding equation yielded 96% correct classification with 90% sensitivity and 100% specificity. The classification performance of any single variable alone was considerably worse. CONCLUSION: Combination of information from different sources may be required for monitoring the adequacy of analgesia during anaesthesia. PMID- 15752390 TI - Peroperative depth of anaesthesia may influence postoperative opioid requirements. AB - BACKGROUND: Studies on monitoring the depth of anaesthesia have shown that with the use of these monitors the peroperative consumption of anaesthetics can be reduced. Studies have also indicated that the peroperative depth of anaesthesia may affect the postoperative course. The purpose of this study was to evaluate a possible relation between the depths of anaesthesia and the postoperative pain score and consumption of morphine. METHODS: We used middle latency auditory evoked potentials (MLAEPs) for monitoring the depth of anaesthesia. The study was prospective, observer blinded and included 50 women scheduled for elective abdominal hysterectomy. Anaesthesia was induced using propofol and remifentanil. Before leaving the recovery room the patients were provided with a programmed patient-controlled pump (PCA), which was only activated on demand. Auditory evoked potentials were registered from just before induction of anaesthesia and during the whole procedure, but the anaesthetist did not have access to the monitor. RESULTS: Fifty patients were included and seven were excluded. The remaining 43 patients were divided into two groups: Group High (n=12) with an AAI>28 for >5% of the registration time and Group Low (n=31) with an AAI>28 for <5% of the registration time. Group High had significantly higher morphine requirements in the recovery and activated the PCA-pump more frequently during the first 24 postoperative hours. CONCLUSION: The results indicate that the peroperative depth of anaesthesia may have effects on the postoperative analgesic requirements. PMID- 15752391 TI - Delayed emergence process does not result in a lower incidence of emergence agitation after sevoflurane anesthesia in children. AB - BACKGROUND: Emergence agitation (EA) is more frequent after sevoflurane anesthesia than other inhalational agents but the etiology remains unclear. We investigated whether the EA after sevoflurane anesthesia is related to rapid emergence. METHODS: Eighty-five patients, aged 1-7 years, undergoing elective urology surgery were studied. After induction, anesthesia was maintained with 2.0 2.5% sevoflurane and 50% N(2)O. BIS was monitored continuously throughout surgery and emergence. At the end of surgery, the children were randomly assigned to one of two groups: group I, immediate cessation of sevoflurane; or group G, gradual decrease of sevoflurane with the rate of 0.1%.min(-1). Emergence agitation score was recorded during recovery. RESULTS: Times from immediate cessation or start of decrease of sevoflurane to BIS 70, 80, 90 and extubation were significantly longer in group G. The incidence of severe agitation (score > or =3) was not significantly different between group I and group G: which were 35.7% and 32.6%, respectively. CONCLUSION: Prolonged recovery did not reduce the incidence of EA following sevoflurane anesthesia in children. PMID- 15752392 TI - Preoperative caudal block prevents emergence agitation in children following sevoflurane anesthesia. AB - BACKGROUND: The frequency of emergence agitation in children is increased following sevoflurane anesthesia. However, controversies still exist concerning the exact etiology of this postanesthetic problem. Although this phenomenon is present with adequate pain relief or even following pain-free procedures, pain is still regarded as a major contributing factor. METHODS: In a prospective, randomized, double-blind study, we enrolled 48 premedicated and calm 2-6-year-old children undergoing inguinal hernia repair. We assigned children to one of two groups: children assigned to the caudal group (n = 24) received a caudal block to supplement sevoflurane, while children assigned to the fentanyl group (n = 24) received a bolus injection of 1 microg kg(-1) intravenous fentanyl before skin incision to supplement sevoflurane. In the post anesthesia care unit, all children were received by their parent, and the incidence of emergence agitation and pain scores, as well as hemodynamic changes, were compared in both groups. RESULTS: Forty-four children completed the study. In the fentanyl group, 59% of the children were agitated following emergence from anesthesia as compared to 4.5% in the caudal group (P < 0.001). Also, pain scores, mean values of heart rate and blood pressure as well as morphine requirement were significantly higher in the post anesthesia care unit in the fentanyl group compared to the caudal group. CONCLUSION: Our results show that in children undergoing inguinal hernia repair, pain control with a preoperative caudal block as compared to intraoperative intravenous fentanyl significantly reduces the incidence of emergence agitation and pain scores following sevoflurane anesthesia. PMID- 15752394 TI - Variability of duration of action of neuromuscular-blocking drugs in elderly patients. AB - BACKGROUND: Steroid-based, non-depolarizing neuromuscular-blocking (NMB) drugs (e.g. rocuronium, vecuronium) are characterized by organ-dependent elimination and significantly longer durations of action in elderly compared to young patients. Cisatracurium is a benzylisoquinolinium NMB drug with a duration of action not altered by ageing. The objective of the study was to determine if elderly patients had less variability in duration of action with 2 x ED95 of cisatracurium compared to equipotent doses of rocuronium or vecuronium. METHODS: Informed consent was obtained from 66 elderly patients with normal renal and liver function. Preoperative midazolam (1 mg) was given IV. The anaesthestic induction was with 5 mg kg(-1) thiopental and 2 microg kg(-1) fentanyl. The patients received 0.6 mg kg(-1) rocuronium, 0.1 mg kg(-1) vecuronium or 0.1 mg kg(-1) cisatracurium. Anaesthetic maintenance was with sevoflurane in oxygen/nitrous oxide. Neuromuscular-blocking duration of action was defined as the return of T1 twitch height to 25% of control. Variability was determined by subtracting the actual duration of action from the mean duration of action for each drug. RESULTS: The durations of action (range, min) were: cisatracurium, 37 81; vecuronium, 35-137; and rocuronium, 33-119. The median of the variability of duration was significantly less with cisatracurium (7 min) compared to vecuronium (18 min) and rocuronium (18 min) (P < 0.05). CONCLUSION: When used with sevoflurane/N(2)O, there was a two-fold greater variability of duration of neuromuscular blockade in elderly patients receiving rocuronium or vecuronium compared with cisatracurium. PMID- 15752393 TI - Less postoperative nausea and vomiting after propofol + remifentanil versus propofol + fentanyl anaesthesia during plastic surgery. AB - BACKGROUND: The effect of different opioids on postoperative nausea and vomiting (PONV) has not been conclusively determined yet, thus the aim of this study was to compare the incidence of PONV in propofol-anaesthetized patients receiving either fentanyl or remifentanil as opioid supplement. METHODS: Sixty ASA physical status I and II patients scheduled for plastic surgery gave their written informed consent for this prospective, randomized, double-blind study. Anaesthesia was induced with propofol, rocuronium and fentanyl (n = 30; 2 microg kg(-1)) or remifentanil (n = 30; 1 microg kg(-1)). After tracheal intubation, anaesthesia was maintained with propofol, oxygen in air and an infusion of the opioid studied, which was modified according to clinical criteria. Baseline postoperative analgesia was achieved with intravenous propacetamol + metamizol. Intravenous morphine was given if visual analogic scale (VAS) for pain was > or = 4 (scale 0-10) and metoclopramide was administered if a patient presented > or = 2 PONV episodes (nausea or vomiting) in less than 30 min. Postoperatively (2, 12 and 24 h), we registered VAS, rescue morphine consumption, number of patients with episodes of PONV and number of patients requiring metoclopramide. P < 0.05 was considered significant. RESULTS: There were no significant differences between groups in the demographic parameters, ASA physical status, propofol dose, VAS, and rescue morphine requirements. Fourteen patients in the fentanyl group and four in the remifentanil group presented PONV episodes 2-12 h postoperative hours' interval; (P < 0.05). Ten patients in the fentanyl group and four in the remifentanil group presented vomiting episodes in the same period (P < 0.05); and eight patients in the fentanyl group and one in the remifentanil group required metoclopramide; (P < 0.05). The number of postoperative PONV episodes were low, both in the 0-2-h period (n = 2 vs. n = 1, fentanyl and remifentanil, respectively) and in the 12-24-h period (n = 3 vs. n = 1). CONCLUSION: Propofol + fentanyl anaesthesia resulted in a higher incidence of PONV and requirements of antiemetic drugs in the period between 2 and 12 postoperative hours compared with propofol + remifentanil, in patients undergoing plastic surgery. PMID- 15752395 TI - Acceleromyography vs. electromyography: an ipsilateral comparison of the indirectly evoked neuromuscular response to train-of-four stimulation. AB - BACKGROUND: There is a considerable body of evidence which suggests that data obtained using acceleromyography (AMG) cannot be used interchangeably with observations obtained by mechanomyographic (MMG) or electromyograhic (EMG) methods. All previous such studies evaluated the responses from contralateral limbs. This investigation was undertaken to determine if these previously described differences were in part a function of observing the responses from opposing limbs. METHODS: We compared the ipsilateral EMG and AMG response to an ED(95) bolus of atracurium in 50 subjects. In half of the individuals the thumb was free to move freely; in half, a small elastic preload was applied to the thumb. Train-of-four (TOF) recovery was followed until a TOF ratio >0.90 was recorded by both monitors. Acceleromyography vs. EMG differences and the resultant 95% confidence limits for twitch height (T1) and the TOF ratio were determined. RESULTS: When the AMG TOF value had recovered to a value of 0.72 +/- 0.03; the simultaneously evoked EMG value averaged only 0.59 +/- 0.08. This difference was statistically significant (P < 0.001). Although the mean difference AMG vs. EMG was little more than 0.10, differences in an individual might be twice that amount. When the AMG TOF value had recovered to 0.90, the simultaneously evoked EMG value averaged 0.85. Again the 95% confidence limits for individual observations was very wide. With EMG, once the TOF ratio returns to a value of 0.70, T1 has returned to 95% of control. In contrast with AMG, return of T1 -95% of control requires a TOF ratio of almost 0.90. Addition of an elastic preload to the thumb decreased control TOF variability without effecting the relationship between twitch height and the TOF ratio. CONCLUSION: Acceleromyographic TOF values tend to overestimate the extent of EMG recovery. Acceleromyographic TOF values <0.90 are indicative of incomplete neuromuscular recovery. PMID- 15752396 TI - Dose-response relationship of rocuronium: a comparison of electromyographic vs. acceleromyographic-derived values. AB - BACKGROUND: Acceleromyography (AMG) is being employed with increasing frequency as a research tool. However, there is almost no information available regarding the accuracy of values for drug potency obtained using AMG. This study was an attempt to determine if AMG-derived ED(50/95) values are interchangeable with those measured with a more traditional neuromuscular monitor. METHODS: Thirty adult patients were studied. Anesthesia was induced and maintained with N20, propofol, and supplementation opioid. Tracheal intubation was accomplished without muscle relaxants. Simultaneous ipsilateral AMG and EMG responses to 0.10 Hz stimulation was recorded. Following instrument calibrations, a single dose of rocuronium was administered. The first patient received a bolus of 0.17 mg kg(-1) of rocuronium. Using the Hill equation with a postulated slope of 4.50, the ED(50) was calculated. The second subject received a dose which approximated the calculated ED(50) for patient no. 1. Successive subjects were given a dose based on the running average of the estimated ED(50). RESULTS: The AMG-derived ED50/95 values for rocuronium (0.163 +/- 0.055 and 0.314 +/- 0.105 mg mg(-1)) were virtually identical to those established using EMG (0.159 +/- 0.043 and 0.306 +/- 0.084 mg kg(-1)). While mean peak twitch depression (Delta T1) was the same in both groups for individual subjects Delta T1 differed by +/- 20% (95% confidence interval). DISCUSSION: Acceleromyography-derived twitch heights for individual patients are not necessarily interchangeable with information obtained using electromyography. Nevertheless, acceleromyography appears to be a valid methodology for determining the drug potency when a population rather than an individual subject is being studied. PMID- 15752397 TI - Increase in the use of rebreathing gas flow systems and in the utilization of low fresh gas flows in Finnish anaesthetic practice from 1995 to 2002. AB - BACKGROUND: The use of rebreathing systems together with low fresh gas flows saves anaesthetic gases, reduces the costs of anaesthesia, causes less environmental and ergonomic adverse effects, i.e. less air contamination in the operating room, and has favourable physiological effects. We assessed whether the use of non-rebreathing vs. rebreathing gas flow systems and high vs. lower fresh gas flows has changed during recent years. METHODS: The use of rebreathing and non-rebreathing systems and the utilization of fresh gas flows were evaluated by sending a questionnaire to the heads of anaesthesia departments at all public health care hospitals in Finland in 1996 and 2003. The data was gathered from the previous years 1995 and 2002, respectively. RESULTS: The use of rebreathing systems increased from 62% to 83% of all instances of general anaesthesia (P < 0.001). In rebreathing gas flow systems, there was a significant shift from high fresh gas flows (3 l min(-1) and more) towards lower fresh gas flows (between 1 to 2 l min(-1) and even below 1 l min(-1)) (P < 0.001). CONCLUSIONS: The benefits of low fresh gas flows have now been achieved in most instances of rebreathing system anaesthesia, which was not the case in 1995. PMID- 15752398 TI - Rapid fluid infusion therapy decreases the plasma concentration of continuously infused propofol. AB - BACKGROUND: Rapid fluid infusion therapy to treat hypovolemia in anesthetized patients is a common practical regimen in daily clinical settings. This study investigated the effect of large volume loading on the plasma concentration of propofol (Cp), hemodynamic parameters, hemoglobin concentration (Hb), hematocrit value (Ht) and the bispectral index (BIS). METHODS: Sixty patients were administered propofol using a target-controlled infusion technique. We studied two independent groups. Half of the patients (group F, n = 30) were administered fentanyl, and the other half (group E, n = 30) epidural administration of mepivacaine for analgesia. After achieving a pseudo-steady state of propofol anesthesia, baseline values of blood pressure, heart rate, Hb, Ht, cardiac output, Cp and BIS were measured, and 10 ml/kg Ringer's solution was infused over 15 min. RESULTS: In group F, Cp was significantly decreased from 2.24 (0.69) [mean (SD)] to 2.07 (0.61) microg/ml and in group E from 2.02 (0.98) to 1.75 (0.51) microg/ml immediately after infusion (P < 0.05). The significant reduction lasted until 30 min in group F, whereas, Cp quickly recovered in group E. Cardiac output was increased only in group F. The dilution ratio demonstrated the prolonged diluting effect in group E and the significant correlation with the rate of decrease in Cp (P < 0.0003, R = 0.21). The BIS value showed no significant change immediately after infusion. CONCLUSION: Large volume loading decreased Cp without a significant change in BIS values. The effect of infusion therapy on the depth of anesthesia might be small and usually negligible during propofol anesthesia. PMID- 15752399 TI - Long-time outcome after transient transtentorial herniation in patients with traumatic brain injury. AB - BACKGROUND: This study investigates mortality and morbidity in patients with traumatic brain injury (TBI) who developed episode(s) of transtentorial herniation. The transtentorial herniation was defined as a deterioration of consciousness accompanied by uni- or bilateral pupil dilatation. METHODS: The medical records of all patients with traumatic brain injury admitted during 1999 to the Neuro- or General Intensive Care Units at Sahlgrenska University Hospital were analyzed, and patients with at least one episode of transtentoryal herniation were included. Information regarding patient age, gender, type of trauma, initial GCS, precipitating reason for herniation, uni-/bilateral pupil dilatation, treatment(s) and outcome after at least 6 months, assessed with the Glasgow Outcome Scale (GOS), was collected from medical records. RESULTS: The study included 27 patients, average age 44 years (range 6-81), with a male proportion of 81%. The majority of the patients were victims of traffic accidents and falls. The results demonstrated that 16/27(59%) of the patients had a favorable outcome (GOS 4/5), 4/27(15%) were severely disabled (GOS 3), none was vegetative (GOS 2) and 7/27(26%) died (GOS 1). When analyzing patient subgroups, best outcome was found in children where 3/4 (75%) had a GOS 4/5. CONCLUSION: Transtentorial herniation is a serious consequence of supratentorial edema/mass lesions in patients with TBI. However, with aggressive neurointensive care and neurosurgical treatments we found a 59% patient incidence of a favorable outcome. PMID- 15752400 TI - Treatment of elevated intracranial pressure with indomethacin: friend or foe? AB - Indomethacin has been suggested as a therapeutic tool to manage elevated intracranial pressure in patients with severe head injury and patients undergoing craniotomy for brain tumors. Indomethacin is a non-selective cyclooxygenase inhibitor. Compared to other cyclooxygenase inhibitors indomethacin has unique effects on cerebral blood flow. Administration of indomethacin causes cerebral vasoconstriction and decreases cerebral blood flow, which elicits a decrease in intracranial pressure. The mechanism of indomethacin-induced cerebral vasoconstriction is not completely understood and controversies exist whether indomethacin causes cerebral ischemia. The primary aims of this article were to review the existing knowledge of indomethacin's influence upon cerebral hemodynamics and elevated ICP in patients with brain pathology. Furthermore, indomethacin's mechanism of action and whether it causes cerebral ischemia are discussed. PMID- 15752401 TI - Mild hypothermia reduces expression of intercellular adhesion molecule-1 (ICAM-1) and the accumulation of neutrophils after acid-induced lung injury in the rat. AB - BACKGROUND: The pathophysiology of the acute phase of acid-induced lung injury (AILI) has been elucidated. However, once acute respiratory distress syndrome (ARDS) develops, the mortality rate remains high and there is, as yet, no effective therapy. There are reports that application of mild hypothermia is an effective treatment for ARDS. In this study, we hypothesize that mild hypothermia inhibits activation of neutrophils and expression of intercellular adhesion molecule-1 (ICAM-1) in an injured lung. We studied the effects of mild hypothermia on the expression of ICAM-1 and the accumulation of neutrophils after AILI in the rat. METHODS: Male Sprague-Dawley rats were randomly allocated to one of the four groups: control normothermic group, induced mild hypothermia group, acid-instilled normothermic group, and acid-instilled group with mild hypothermia. At 6 h after instillation of acid, lungs were removed to measure neutrophil activity and to detect the expression of ICAM-1 in each group. RESULTS: Oxygenation in acid-instilled rats was significantly impaired as compared to that in non-instilled groups, but induction of mild hypothermia gradually improved oxygenation. Expression of ICAM-1 was enhanced in the acid instilled normothermic group. By contrast, no overexpression of ICAM-1 and its mRNA was detected in the acid-instilled hypothermic group. In addition, accumulation of neutrophils was markedly inhibited after exposure to mild hypothermia irrespective of the instillation of acid. CONCLUSION: Our data suggest mild hypothermia can inhibit the adhesion, activation, and accumulation of neutrophils during the acute phase of AILI in the rat and may have the potential to reduce ongoing inflammation of ALI or ARDS. PMID- 15752402 TI - Are there changes in leg vascular resistance during laparoscopic cholecystectomy with CO2 pneumoperitoneum? AB - BACKGROUND: The prompt haemodynamic response to carbon dioxide insufflation during laparoscopic cholecystectomy suggests involvement of the sympathetic system. The aim of the present study was to examine if a change in vascular resistance in leg skeletal muscle could be an important mechanism behind the increased afterload. Furthermore, the arterio-venous differences of the catecholamines were measured in the leg before and during insufflation of carbon dioxide into the peritoneal cavity. METHODS: Ten patients (ASA I) scheduled for laparoscopic cholecystectomy were included. After induction of anaesthesia, catheters were introduced percutaneously into the radial artery, the femoral vein and the cubital vein for pressure monitoring and blood sampling. The arterial blood flow in the legs was measured by mercury-in-Silastic strain gauge venous occlusion plethysmography. Vascular resistance in the right leg (LVR) was calculated from the formula: (MAP-FVP)/calf blood flow. Measurements were made before and 5 min after insufflation of pneumoperitoneum. RESULTS: Induction of pneumoperitoneum increased the heart rate (P < 0.05) and also increased mean arterial pressure and femoral vein pressure as well as the calculated leg vascular resistance (P < 0.01). Calf blood flow did not change significantly in either leg. Both arterial and venous noradrenaline concentrations were higher after insufflation (P < 0.01). CONCLUSION: In patients without heart or lung disease, pneumoperitoneum at an intra-abdominal pressure level of 11-13 mmHg increased the peripheral vascular resistance in the leg while the arterial blood flow in the leg was unaffected. Catecholamine levels increased, but were still low. Therefore, we suggest that the increase in peripheral vascular resistance is caused by increased myogenic activity in the resistance vessels secondary to increased arterial and transmural pressure rather than by increased neurogenic sympathetic activity. PMID- 15752403 TI - Precision of bolus thermodilution cardiac output measurements in patients with atrial fibrillation. AB - BACKGROUND: The precision of bolus thermodilution cardiac output measurements in patients with atrial fibrillation (AF) has not previously been determined. A priori we suspected that the precision would be lower in patients with AF than in patients with sinus rhythm (SR). Consequently, we also determined if the precision could be improved by injecting the thermal indicator into the right ventricle instead of the right atrium. METHODS: Cardiac output was determined as the average result of four injections of 10 ml of iced saline. Replicate measurements were performed with thermal indicator injections into the right atrium and ventricle. The coefficients of variation and the precisions were calculated. RESULTS: In the 25 patients with AF, mean cardiac output was 3.96 l min(-1) (range 2.4-7.4), the coefficient of variation 0.073 (95% CI +/- 0.011), and the precision 0.38 l min(-1) (95% CI +/- 0.14) with injection into the right atrium. In the 25 patients with SR, mean cardiac output was 4.73 l min(-1) (range 2.4-7.3), the coefficient of variation 0.047(95% CI +/- 0.006), and the precision 0.38 l min(-1) (95% CI +/- 0.14). In both groups, an agreement analysis demonstrated that the injection of indicator into the right ventricle resulted in a significantly higher cardiac output [AF+0.25 (95% CI +/- 0.15) l min(-1), SR+0.29 ( +/- 0.20) l min(-1)]. CONCLUSION: The coefficient of variation for cardiac output determinations is 55% higher in patients with AF. Two measurements, separated by time or intervention, must differ by 15% in AF patients and 9% in SR patients before one can be 95% confident that a real change has taken place. PMID- 15752404 TI - Artificial colloids impair haemostasis. An in vitro study using thromboelastometry coagulation analysis. AB - BACKGROUND: Hydroxyethyl starch (HES) solutions impair haemostatic mechanisms. The impact of the degree of substitution (DS) of a HES solution on thromboelastometry tracings is unclear. Therefore we tested the hypothesis of whether the DS has an effect on the haemostatic defect caused by HES, and assessed whole blood coagulation by thromboelastometry coagulation analysis (ROTEM, Pentapharm Co., Munich, Germany) in serial in vitro haemodilutions of colloids. METHODS: Whole blood was withdrawn from 12 volunteers in a crossover study. Six per cent low-molecular weight HES with a high (HES MW 120 kDa/degree of substitution 0.7) and low (HES MW 130 kDa/0.4) degree of substitution, 4% succinylated gelatin (GEL) or 4% albumin (ALB) was added to citrated venous whole blood samples to make 20, 40, 60 vol.% end-concentrations of each of the solutions. Samples were analyzed by ROTEM. RESULTS: There was a comparable decrease in maximum clot firmness (MCF) and shear elastic modulus [G = 5000 x MCF/(100-MCF)] by HES 120/0.7 and HES 130/0.4 at 20 and 40 vol.% dilutions. At 60 vol.% dilution HES 120/0.7 decreased less alpha-angle and MCF than HES 130/0.4 (P < 0.05). With moderate dilutions all colloids shortened coagulation time (CT). At 20, 40 and 60 vol.% dilutions MCF and G were more decreased in both HES groups than in the ALB and GEL groups (P < 0.05). Furthermore, at 40 and 60 vol.% dilutions G deteriorated more in the GEL than in the ALB group (P < 0.05). CONCLUSION: In vitro the impact of the degree of substitution of HES solution on thromboelastometry coagulation analysis was modest. Haemodilution with gelatin and albumin induced fewer coagulation abnormalities than HES. In addition, the haemodilution with gelatin impaired coagulation more than albumin solution. PMID- 15752405 TI - Vasodilatory shock in severe acute pancreatitis without sepsis: is there any place for hydrocortisone treatment? AB - BACKGROUND: Hydrocortisone (HC) has been reported to rapidly improve hemodynamics and reduce the time to vasopressor cessation in septic shock, but none has focused on this effect in acute pancreatitis. We therefore performed a study to assess the effects of hydrocortisone on catecholamine-dependent shock among patients with severe acute pancreatitis. METHODS: A retrospective, case controlled study among 10 patients with severe acute pancreatitis and HC treatment for catecholamine-dependent shock was performed. The control group comprised 11 conventionally treated patients with the same severity of pancreatitis and circulatory shock according to the norepinephrine support required. In focus were the first 48 h from the start of HC administration in the HC group and from the reference point in the control group, respectively. The reference point for the control group was the time point at which doses of norepinephrine exceeded 0.3 microg kg(-1) min(-1). RESULTS: Patients in the HC group were weaned off norepinephrine in a significantly shorter time (61 h in HC group vs. 141 h, P = 0.016). The HC group received significantly less norepinephrine (area under curve of norepinephrine dose, P = 0.041). The reduction in norepinephrine dose was comparable at 24 h, being -0.051 (-0.208 0.022) microg kg(-1) min(-1) in the HC group vs. -0.026 (-0.150-0.030) microg kg( 1) min(-1) in the controls (P = 0.307), and at 48 h with respective figures of 0.206 (-0.317 to -0.102) microg kg(-1) min(-1) and -0.103 (-0.178-0.029) microg kg(-1) min(-1) (P = 0.072), from the start of HC administration. CONCLUSION: According to our data it seems reasonable to formulate a hypothesis that low doses of HC shorten the time to vasopressor cessation and rapidly reduce the need for norepinephrine support in patients with shock associated with severe acute pancreatitis without sepsis. PMID- 15752406 TI - Effects of edaravone on human neutrophil function. AB - BACKGROUND: Neutrophils play a crucial role in the antibacterial host defence system. Edaravone is used in critically ill patients who are often immuno compromised secondary to concomitant disease or immunosuppressive therapy. The aim of the current study was to assess the effect of edaravone, a novel free radical scavenger, on several aspects of human neutrophil function using an in vitro system. METHODS: Chemotaxis, phagocytosis, reactive oxygen species (ROS) production by neutrophil (cellular) and xanthine-xanthine oxidase (acellular) systems, and intracellular calcium ion levels ([Ca(2 +) ]i) were measured in the absence and in the presence (at a clinically relevant concentration, and 0.1 fold, and 10-fold this concentration) of edaravone. RESULTS: The clinically relevant concentration of edaravone did not inhibit chemotxais, phagocytosis, or superoxide production of neutrophils. Even at its ordinary clinical plasma concentration, the drug inhibited hydrogen peroxide (H(2)O(2)) and hydroxyl radical (OH*) generation in the cellular (neutrophil) as well as in the cell-free (xanthine-xanthine oxidase) system (P < 0.05). Edaravone did not affect elevation of [Ca(2 +) ]i in neutrophils stimulated by a chemotactic factor. CONCLUSIONS: These findings suggest that edaravone quenched H(2)O(2), and OH* generated rather than impaired the ability of neutrophils to produce the ROS. However, further studies using in vivo systems are required to elucidate the effects of edaravone on neutrophil function in clinical settings. PMID- 15752407 TI - Lack of effect of ranitidine on gastric luminal pH and mucosal PCO2 during the first day in the ICU. AB - BACKGROUND: Histamine(2) (H(2))-blocking agents can attenuate intragastric CO(2) production by reducing gastric acid secretion and preventing the interaction between H(+) and bicarbonate. However, gastric acid production may be impaired in acute circulatory failure due to poor mucosal perfusion, and H(2)-blockade could further impair mucosal perfusion. METHODS: Forty patients with acute circulatory and/or respiratory failure, age 61 +/- 16 years (mean +/- SD), APACHE II score 21 +/- 7, and SOFA score 8 +/- 3, received randomly either ranitidine, 50 mg (R) or placebo (P) every 8 h. Gastric intraluminal pH (gpH; antimony probe with external reference electrode) and mucosal pCO(2) (prCO(2), semicontinuous air-tonometry) were measured during 24 h, and blood gases were taken at 6-h intervals. RESULTS: Gastric intraluminal pH was 4.3 +/- 2.4 in P and 5.1 +/- 1.6 in R (NS). Mean prCO(2) was 6.8 +/- 2.7 kPa in P and 7.4 +/- 2.1 kPa in R, and mucosal-arterial pCO(2) gradient (Delta pCO(2)) was 2.2 +/- 2.9 kPa and 2.4 +/- 2.4 kPa, respectively (NS). Within-patient variabilities of gpH and prCO(2) were not influenced by ranitidine. A posthoc analysis revealed that non-survival in R was associated with a low mucosal pHi after 24 h (P = 0.002). This was explained by a low arterial pH but not by differences in gpH or prCO(2). CONCLUSION: In acute respiratory and circulatory failure, H(2) blockade has an inconsistent impact on gpH and does not reduce variabilities of gpH or prCO(2). PMID- 15752408 TI - Pharmacokinetics of 0.5% levobupivacaine following ilioinguinal-iliohypogastric nerve blockade in children. AB - BACKGROUND: Bupivacaine (2 mg kg(-1)) has been recommended for blockade of the ilioinguinal and iliohypogastric nerves in paediatric patients undergoing inguinal surgery. We determined the plasma concentrations of levobupivacaine following ilioinguinal-iliohypogastric block. METHODS: Twenty children scheduled for elective surgery for inguinal surgery received 2 mg kg(-1) of 0.5% levobupivacaine. Surgical anaesthesia was maintained with mask inhalation of oxygen, nitrous oxide and sevoflurane. Venous blood samples were drawn at regular intervals up to 2 h and plasma was separated. Total venous plasma concentrations were determined by gas chromatography. Bupivacaine concentrations from a study with a similar protocol were used as historical controls for comparison. RESULTS: The groups were similar with respect to age, weight and dosage of local anaesthetic. The initial distribution half-time (Talpha), the peak plasma concentration (Cmax) achieved, the time to the peak plasma concentration were similar (Tmax), and the mean areas under the concentration time curve (AUC) were similar between the two local anaesthetics. CONCLUSIONS: Levobupivacaine and bupivacaine are equally absorbed to similar maximum concentrations. PMID- 15752409 TI - Intrathecal fentanyl added to hyperbaric ropivacaine for transurethral resection of the prostate. AB - BACKGROUND: Our purpose was to evaluate the effect of intrathecal fentanyl 25 microg added to 18 mg of 6 mg ml(-1) hyperbaric ropivacaine on the characteristics of subarachnoid block and postoperative pain relief in patients undergoing TURP surgery. METHODS: The patients were randomly assigned into two groups: Group S (saline group, n=16) received 3 ml of 18 mg hyperbaric ropivacaine + 0.5 ml saline--in total, a 3.5-ml volume intrathecally; and Group F (fentanyl group, n=15) received 3 ml of 18 mg hyperbaric ropivacaine + 0.5 ml of 25 microg fentanyl--in total, a 3.5-ml volume intrathecally. In both groups the onset and recovery times of the sensory block, degree and recovery times of the motor block and side-effects were recorded and statistically compared. RESULTS: There was no significant difference between the groups in achieving the highest level of sensory block, and in the times taken to reach the peak level. Regression to L1 was significantly prolonged in the fentanyl group compared with the saline group (P=0.004). Times to the first feeling of pain and the first analgesic requirement were significantly prolonged in the fentanyl group compared with the saline group (P=0.011 and P=0.016, respectively). The frequency of pruritus was significantly higher in the fentanyl group compared with the saline group (P=0.022). CONCLUSION: Addition of fentanyl 25 microg to hyperbaric ropivacaine 18 mg for spinal anesthesia in patients undergoing TURP may significantly improve the quality and prolong the duration of analgesia, without causing a substantial increase in the frequency of major side-effects. PMID- 15752410 TI - Anesthetic effect of epidural anesthesia with cephalad or caudad catheterization for ankle surgery or hemorrhoidectomy. AB - BACKGROUND: The larger size of the first sacral nerve root has been reported to be an unfavorable factor leading to sacral sparing in epidural anesthesia. Previous studies have shown that an adequate analgesic effect of the epidural block was achieved with the catheter placement in the caudal direction. In this study, the anesthetic effect of epidural anesthesia with catheter placement of a cephalic or caudad direction was compared in ankle and hemorrhoid surgery. METHODS: Twenty-one ASA physical status I or II patients undergoing surgery for ankle fractures with epidural anesthesia were enrolled and randomized into two groups. The epidural catheter was placed either to a cephalad (AU group) or caudal (AD group) direction. Another 21 patients undergoing hemorrhoidectomy were also randomized into two groups to receive epidural anesthesia in a similar way (HU and HD groups). The onset for, duration of, and recovery time from epidural anesthesia and the incidence of analgesic request were recorded. RESULTS: No significant differences were demonstrated when age, height, weight or sex were compared between the four study groups. The onset time of the block and the incidence of intrasurgical analgesic request were lower in the caudal subgroup when the ankle surgery patients were compared. Otherwise, there were no significant differences in the duration of anesthesia and time to recovery or level of anesthesia. CONCLUSION: Injection of local anesthetic solution through a caudally oriented epidural catheter produces faster onset and superior quality of anesthesia in comparison with the injection through the cephaladly oriented catheter in ankle surgery, but not hemorrhoidectomy. PMID- 15752411 TI - Intensive care treatment of severe mixed metabolic acidosis. AB - We report a case of severe metabolic acidosis associated with acute renal failure and septicaemia following treatment with maximal therapeutic doses of metformin and diclofenac. On the second day of intensive care the patient deteriorated with respiratory insufficiency and abdominal pain during continuous renal replacement therapy. A laparoscopy revealed a perforated cholecystitis with abscess formation. The patient regained renal function and recovered. Intake of diclofenac 5 days before this episode could have been the main cause of renal insufficiency and metabolic acidosis in this patient and could also have delayed surgical treatment by masking early clinical signs of perforated cholecystitis. The renal failure may also have caused metformin and lactate to accumulate, contributing to the mixed pattern of metabolic acidosis. This case report describes a mixed organic and non-organic metabolic acidosis associated with acute renal failure, presumably resulting from a combination of drugs and diseases often found in the elderly - metformin for diabetes mellitus and a non steroidal anti-inflammatory drug for cholecystolithiasis. Acid-base balance and electrolyte changes were rapidly normalized by continuous renal replacement therapy. PMID- 15752412 TI - Nitric oxide inhalation is useful in the management of right ventricular failure caused by myocardial contusion. AB - We report a severe head injury and blunt chest trauma with sternal and multiple rib fractures with high-energy impact in a 22-year-old man. Twelve hours after the accident, haemodynamic status of the patient rapidly worsened because of right ventricular (RV) failure due to myocardial contusion, requiring increasing doses of catecholamine. Nitric oxide inhalation was used to decrease RV afterload, and produced an immediate improvement in haemodynamic status, permitting a decrease in catecholamine administration. From days 2 through 8, cardiac function continued to improve, and was normal on day 9. Nitric oxide inhalation was stopped on day 4. PMID- 15752413 TI - General anesthesia with remifentanil for Cesarean section in a patient with HELLP syndrome. AB - HELLP syndrome is a severe complication of pre-eclampsia characterised by hemolysis, elevated liver enzymes and a low platelet count. It is associated with an increased risk of adverse outcome for both the mother and the fetus. Patients with HELLP syndrome are also at greater risk of pulmonary edema, adult respiratory distress syndrome, abruptio placentae, intracerebral hemorrhage, eclamptic convulsions, disseminated intravascular coagulation, ruptured liver hematomas and acute renal failure. Perinatal mortality is equally high. Before delivery, aggressive obstetric management is directed toward stabilization of the affected organ systems, if possible, and interruption of the pregnancy in the early phase of the accelerated disease progression. Definitive therapy is delivery. Parturients HELLP syndrome often require general anesthesia for Cesarean section delivery. The anesthetic technique is critical for these patients with a high risk of uncontrollable hypertension, bleeding and multiple organ failure. Remifentanil is increasingly used as a very short analgesic agent providing cardiovascular stability in high-risk patients. We report the management of a patient presenting in labor with HELLP syndrome, and describe the successful use of remifentanil as part of the anesthetic technique for her subsequent Cesarean section. PMID- 15752414 TI - Use of a new supraglottic airway device--the CobraPLA--in a 'difficult to intubate/difficult to ventilate' scenario. AB - A 67-year-old male was scheduled for cadaveric renal transplant. Following rapid sequence anaesthesia, only the tip of the epiglottis was visualized. Mask ventilation was difficult, and therefore an alternative airway device was considered. A size 4 CobraPLA (Engineered Medical System, Indianapolis, IN) was inserted into the hypopharynx, and ventilation was easily accomplished. Then, a size 7 endotracheal tube was threaded over a fiberoptic bronchoscope, through the CobraPLA, into the trachea. The description and insertion technique of the CobraPLA, a new supraglottic airway device is shortly discussed. This is the first report of successful management of difficult mask ventilation with the new CobraPLA, supraglottic airway device. PMID- 15752415 TI - The safety of the perioperative administration of cyclooxygenase-2 inhibitors for post-surgical pain. PMID- 15752416 TI - Propofol and questionable lack of sedation. PMID- 15752418 TI - Subdural empyema due to spinal anesthesia. PMID- 15752419 TI - Pk value does depend on the fineness of the observer scale. PMID- 15752421 TI - Using item response theory to explore the psychometric properties of extended matching questions examination in undergraduate medical education. AB - BACKGROUND: As assessment has been shown to direct learning, it is critical that the examinations developed to test clinical competence in medical undergraduates are valid and reliable. The use of extended matching questions (EMQ) has been advocated to overcome some of the criticisms of using multiple-choice questions to test factual and applied knowledge. METHODS: We analysed the results from the Extended Matching Questions Examination taken by 4th year undergraduate medical students in the academic year 2001 to 2002. Rasch analysis was used to examine whether the set of questions used in the examination mapped on to a unidimensional scale, the degree of difficulty of questions within and between the various medical and surgical specialties and the pattern of responses within individual questions to assess the impact of the distractor options. RESULTS: Analysis of a subset of items and of the full examination demonstrated internal construct validity and the absence of bias on the majority of questions. Three main patterns of response selection were identified. CONCLUSION: Modern psychometric methods based upon the work of Rasch provide a useful approach to the calibration and analysis of EMQ undergraduate medical assessments. The approach allows for a formal test of the unidimensionality of the questions and thus the validity of the summed score. Given the metric calibration which follows fit to the model, it also allows for the establishment of items banks to facilitate continuity and equity in exam standards. PMID- 15752422 TI - Phylogenomic approaches to common problems encountered in the analysis of low copy repeats: the sulfotransferase 1A gene family example. AB - BACKGROUND: Blocks of duplicated genomic DNA sequence longer than 1000 base pairs are known as low copy repeats (LCRs). Identified by their sequence similarity, LCRs are abundant in the human genome, and are interesting because they may represent recent adaptive events, or potential future adaptive opportunities within the human lineage. Sequence analysis tools are needed, however, to decide whether these interpretations are likely, whether a particular set of LCRs represents nearly neutral drift creating junk DNA, or whether the appearance of LCRs reflects assembly error. Here we investigate an LCR family containing the sulfotransferase (SULT) 1A genes involved in drug metabolism, cancer, hormone regulation, and neurotransmitter biology as a first step for defining the problems that those tools must manage. RESULTS: Sequence analysis here identified a fourth sulfotransferase gene, which may be transcriptionally active, located on human chromosome 16. Four regions of genomic sequence containing the four human SULT1A paralogs defined a new LCR family. The stem hominoid SULT1A progenitor locus was identified by comparative genomics involving complete human and rodent genomes, and a draft chimpanzee genome. SULT1A expansion in hominoid genomes was followed by positive selection acting on specific protein sites. This episode of adaptive evolution appears to be responsible for the dopamine sulfonation function of some SULT enzymes. Each of the conclusions that this bioinformatic analysis generated using data that has uncertain reliability (such as that from the chimpanzee genome sequencing project) has been confirmed experimentally or by a "finished" chromosome 16 assembly, both of which were published after the submission of this manuscript. CONCLUSION: SULT1A genes expanded from one to four copies in hominoids during intra-chromosomal LCR duplications, including (apparently) one after the divergence of chimpanzees and humans. Thus, LCRs may provide a means for amplifying genes (and other genetic elements) that are adaptively useful. Being located on and among LCRs, however, could make the human SULT1A genes susceptible to further duplications or deletions resulting in 'genomic diseases' for some individuals. Pharmacogenomic studies of SULT1Asingle nucleotide polymorphisms, therefore, should also consider examining SULT1A copy number variability when searching for genotype-phenotype associations. The latest duplication is, however, only a substantiated hypothesis; an alternative explanation, disfavored by the majority of evidence, is that the duplication is an artifact of incorrect genome assembly. PMID- 15752424 TI - A new Apicomplexa-specific protein kinase family: multiple members in Plasmodium falciparum, all with an export signature. AB - BACKGROUND: Malaria caused by protozoan parasites of the genus Plasmodium spp. is a major health burden in tropical countries. The development of new control tools, including vaccines and drugs, is urgently needed. The availability of genome sequences from several malaria parasite species provides a basis on which to identify new potential intervention targets. Database mining for orthologs to the Plasmodium falciparum trophozoite protein R45, a vaccine candidate, led us identify a new gene family. RESULTS: Orthologs to the P. falciparum trophozoite protein R45 were detected exclusively in protozoan parasites of the phylum Apicomplexa, including several Plasmodium spp., Toxoplasma gondii and Cryptosporidium parvum. All family members are hybrid genes with a conserved C terminal protein kinase domain of a novel type, recently called FIKK kinase, associated with a non conserved N-terminal region without any known functional signature. While a single copy gene was detected in most species, considerable gene expansion was observed in P. falciparum and its closest phylogenic relative P. reichenowi, with 20 and six copies, respectively, each with a distinct N terminal domain. Based on full length protein sequence, pairs of orthologs were observed in closely related species, such as P. berghei and P.y. yoelii, P. vivax and P. knowlesi, or P. reichenowi and P. falciparum. All 20 P. falciparum paralogs possess a canonical Plasmodium export element downstream of a signal / anchor sequence required for exportation outside the parasitophorous vacuole. This is consistent with the reported association of the trophozoite protein R45, the only paralog characterised to date, with the infected red blood cell membrane. Interestingly, most genes are located in the subtelomeric region of chromosomes, in association with other multigene families contributing to the remodelling of the infected red blood cell membrane, in particular the ring erythrocyte surface antigen multigene family. CONCLUSION: This Apicomplexan specific gene family was called R45-FIKK kinase. The family hallmark is a kinase domain with unusual characteristics, raising the possibility of designing drug or vaccine strategies targeting this domain. The characteristics of the P. falciparum family suggest a role in remodelling the infected cell and as such possibly contribute to the particular virulence of this species. PMID- 15752423 TI - Emergency vaccination of rabies under limited resources -- combating or containing? AB - BACKGROUND: Rabies is the most important viral zoonosis from a global perspective. Worldwide efforts to combat the disease by oral vaccination of reservoirs have managed to eradicate wildlife rabies in large areas of central Europe and North-America. Thus, repeated vaccination has been discontinued recently on a geographical scale. However, as rabies has not yet been eradicated globally, a serious risk of re-introduction remains. What is the best spatial design for an emergency vaccination program -- particularly if resources are limited? Either, we treat a circular area around the detected case and run the risk of infected hosts leaving the limited control area, because a sufficient immunisation level has not yet been built up. Or, initially concentrate the SAME resources in order to establish a protective ring which is more distant from the infected local area, and which then holds out against the challenge of the approaching epidemic. METHODS: We developed a simulation model to contrast the two strategies for emergency vaccination. The spatial-explicit model is based on fox group home-ranges, which facilitates the simulation of rabies spread to larger areas relevant to management. We used individual-based fox groups to follow up the effects of vaccination in a detailed manner. Thus, regionally -- bait distribution orientates itself to standard schemes of oral immunisation programs and locally -- baits are assigned to individual foxes. RESULTS: Surprisingly, putting the controlled area ring-like around the outbreak does not outperform the circular area of the same size centred on the outbreak. Only during the very first baitings, does the ring area result in fewer breakouts. But then as rabies is eliminated within the circle area, the respective ring area fails, due to the non-controlled inner part. We attempt to take advantage of the initially fewer breakouts beyond the ring when applying a mixed strategy. Therefore, after a certain number of baitings, the area under control was increased for both strategies towards the same larger circular area. The circle circle strategy still outperforms the ring-circle strategy and analysis of the spatial-temporal disease spread reveals why: improving control efficacy by means of a mixed strategy is impossible in the field, due to the build-up time of population immunity. CONCLUSION: For practical emergency management of a new outbreak of rabies, the ring-like application of oral vaccination is not a favourable strategy at all. Even if initial resources are substantially low and there is a serious risk of rabies cases outside the limited control area, our results suggest circular application instead of ring vaccination. PMID- 15752425 TI - cAMP controls cytosolic Ca2+ levels in Dictyostelium discoideum. AB - BACKGROUND: Differentiating Dictyostelium discoideum amoebae respond upon cAMP stimulation with an increase in the cytosolic free Ca2+ concentration ([Ca2+]i) that is composed of liberation of stored Ca2+ and extracellular Ca2+-influx. In this study we investigated whether intracellular cAMP is involved in the control of [Ca2+]i. RESULTS: We analyzed Ca2+-fluxes in a mutant that is devoid of the main cAMP-phosphodiesterase (PDE) RegA and displays an altered cAMP metabolism. In suspensions of developing cells cAMP-activated influx of extracellular Ca2+ was reduced as compared to wild type. Yet, single cell [Ca2+]i-imaging of regA- amoebae revealed a cAMP-induced [Ca2+]i increase even in the absence of extracellular Ca2+. The cytosolic presence of the cAMP PDE inhibitor 3-isobutyl-1 methylxanthine (IBMX) induced elevated basal [Ca2+]i in both, mutant and wild type cells. Under this condition wild type cells displayed cAMP-activated [Ca2+]i transients also in nominally Ca2+-free medium. In the mutant strain the amplitude of light scattering oscillations and of accompanying cAMP oscillations were strongly reduced to almost basal levels. In addition, chemotactic performance during challenge with a cAMP-filled glass capillary was altered by EGTA incubation. Cells were more sensitive to EGTA treatment than wild type: already at 2 mM EGTA only small pseudopods were extended and chemotactic speed was reduced. CONCLUSION: We conclude that there is a link between the second messengers cAMP and Ca2+. cAMP-dependent protein kinase (PKA) could provide for this link as a membrane-permeable PKA-activator also increased basal [Ca2+]i of regA- cells. Intracellular cAMP levels control [Ca2+]i by regulating Ca2+-fluxes of stores which in turn affect Ca2+-influx, light scattering oscillations and chemotactic performance. PMID- 15752426 TI - Genome-scale reconstruction of the metabolic network in Staphylococcus aureus N315: an initial draft to the two-dimensional annotation. AB - BACKGROUND: Several strains of bacteria have sequenced and annotated genomes, which have been used in conjunction with biochemical and physiological data to reconstruct genome-scale metabolic networks. Such reconstruction amounts to a two dimensional annotation of the genome. These networks have been analyzed with a constraint-based formalism and a variety of biologically meaningful results have emerged. Staphylococcus aureus is a pathogenic bacterium that has evolved resistance to many antibiotics, representing a significant health care concern. We present the first manually curated elementally and charge balanced genome scale reconstruction and model of S. aureus' metabolic networks and compute some of its properties. RESULTS: We reconstructed a genome-scale metabolic network of S. aureus strain N315. This reconstruction, termed iSB619, consists of 619 genes that catalyze 640 metabolic reactions. For 91% of the reactions, open reading frames are explicitly linked to proteins and to the reaction. All but three of the metabolic reactions are both charge and elementally balanced. The reaction list is the most complete to date for this pathogen. When the capabilities of the reconstructed network were analyzed in the context of maximal growth, we formed hypotheses regarding growth requirements, the efficiency of growth on different carbon sources, and potential drug targets. These hypotheses can be tested experimentally and the data gathered can be used to improve subsequent versions of the reconstruction. CONCLUSION: iSB619 represents comprehensive biochemically and genetically structured information about the metabolism of S. aureus to date. The reconstructed metabolic network can be used to predict cellular phenotypes and thus advance our understanding of a troublesome pathogen. PMID- 15752427 TI - Could flies explain the elusive epidemiology of campylobacteriosis? AB - BACKGROUND: Unlike salmonellosis with well-known routes of transmission, the epidemiology of campylobacteriosis is still largely unclear. Known risk factors such as ingestion of contaminated food and water, direct contact with infected animals and outdoor swimming could at most only explain half the recorded cases. DISCUSSION: We put forward the hypothesis that flies play a more important role in the transmission of the bacteria, than has previously been recognized. Factors supporting this hypothesis are: 1) the low infective dose of Campylobacter; 2) the ability of flies to function as mechanical vectors; 3) a ubiquitous presence of the bacteria in the environment; 4) a seasonality of the disease with summer peaks in temperate regions and a more evenly distribution over the year in the tropics; 5) an age pattern for campylobacteriosis in western travellers to the tropics suggesting other routes of transmission than food or water; and finally 6) very few family clusters. SUMMARY: All the evidence in favour of the fly hypothesis is circumstantial and there may be alternative explanations to each of the findings supporting the hypothesis. However, in the absence of alternative explanations that could give better clues to the evasive epidemiology of Campylobacter infection, we believe it would be unwise to rule out flies as important mechanical vectors also of this disease. PMID- 15752428 TI - Fuzzy species among recombinogenic bacteria. AB - BACKGROUND: It is a matter of ongoing debate whether a universal species concept is possible for bacteria. Indeed, it is not clear whether closely related isolates of bacteria typically form discrete genotypic clusters that can be assigned as species. The most challenging test of whether species can be clearly delineated is provided by analysis of large populations of closely-related, highly recombinogenic, bacteria that colonise the same body site. We have used concatenated sequences of seven house-keeping loci from 770 strains of 11 named Neisseria species, and phylogenetic trees, to investigate whether genotypic clusters can be resolved among these recombinogenic bacteria and, if so, the extent to which they correspond to named species. RESULTS: Alleles at individual loci were widely distributed among the named species but this distorting effect of recombination was largely buffered by using concatenated sequences, which resolved clusters corresponding to the three species most numerous in the sample, N. meningitidis, N. lactamica and N. gonorrhoeae. A few isolates arose from the branch that separated N. meningitidis from N. lactamica leading us to describe these species as 'fuzzy'. CONCLUSION: A multilocus approach using large samples of closely related isolates delineates species even in the highly recombinogenic human Neisseria where individual loci are inadequate for the task. This approach should be applied by taxonomists to large samples of other groups of closely related bacteria, and especially to those where species delineation has historically been difficult, to determine whether genotypic clusters can be delineated, and to guide the definition of species. PMID- 15752429 TI - A HEV-restricted sulfotransferase is expressed in rheumatoid arthritis synovium and is induced by lymphotoxin-alpha/beta and TNF-alpha in cultured endothelial cells. AB - BACKGROUND: The recruitment of lymphocytes to secondary lymphoid organs relies on interactions of circulating cells with high endothelial venules (HEV). HEV are exclusive to these organs under physiological conditions, but they can develop in chronically-inflamed tissues. The interaction of L-selectin on lymphocytes with sulfated glycoprotein ligands on HEV results in lymphocyte rolling, which represents the initial step in lymphocyte homing. HEV expression of GlcNAc6ST-2 (also known as HEC-GlcNAc6ST, GST-3, LSST or CHST4), an HEV-restricted sulfotransferase, is essential for the elaboration of L-selectin functional ligands as well as a critical epitope recognized by MECA-79 mAb. RESULTS: We examined the expression of GlcNAc6ST-2 in relationship to the MECA-79 epitope in rheumatoid arthritis (RA) synovial vessels. Expression of GlcNAc6ST-2 was specific to RA synovial tissues as compared to osteoarthritis synovial tissues and localized to endothelial cells of HEV-like vessels and small flat-walled vessels. Double MECA-79 and GlcNAc6ST-2 staining showed colocalization of the MECA-79 epitope and GlcNAc6ST-2. We further found that both TNF-alpha and lymphotoxin-alphabeta induced GlcNAc6ST-2 mRNA and protein in cultured human umbilical vein endothelial cells. CONCLUSION: These observations demonstrate that GlcNAc6ST-2 is induced in RA vessels and provide potential cytokine pathways for its induction. GlcNAc6ST-2 is a novel marker of activated vessels within RA ectopic lymphoid aggregates. This enzyme represents a potential therapeutic target for RA. PMID- 15752430 TI - Inclusion of Scar/WAVE3 in a similar complex to Scar/WAVE1 and 2. AB - BACKGROUND: The Scar/WAVE family of proteins mediates signals to actin assembly by direct activation of the Arp2/3 complex. These proteins have been characterised as major regulators of lamellipodia formation downstream of Rac activation and as members of large protein complexes. RESULTS: We have investigated the interactions of the three human Scar/WAVE isoforms with several previously described binding partners for Scar/WAVE 1 or 2. We find that all three Scar/WAVE isoforms behave similarly and are likely to participate in the same kinds of protein complexes that regulate actin assembly. CONCLUSION: Differences between Scar/WAVE proteins are therefore likely to be at the level of tissue distribution or subtle differences in the affinity for specific binding partners. PMID- 15752431 TI - Association between a variation in the phosphodiesterase 4D gene and bone mineral density. AB - BACKGROUND: Fragility fractures caused by osteoporosis are a major cause of morbidity and mortality in aging populations. Bone mineral density (BMD) is a useful surrogate marker for risk of fracture and is a highly heritable trait. The genetic variants underlying this genetic contribution are largely unknown. METHODS: We performed a large-scale association study investigating more than 25,000 single nucleotide polymorphisms (SNPs) located within 16,000 genes. Allele frequencies were estimated in contrasting DNA pools from white females selected for low (<0.87 g/cm2, n = 319) and high (> 1.11 g/cm2, n = 321) BMD at the lumbar spine. Significant findings were verified in two additional sample collections. RESULTS: Based on allele frequency differences between DNA pools and subsequent individual genotyping, one of the candidate loci indicated was the phosphodiesterase 4D (PDE4D) gene region on chromosome 5q12. We subsequently tested the marker SNP, rs1498608, in a second sample of 138 white females with low (<0.91 g/cm2) and 138 females with high (>1.04 g/cm2) lumbar spine BMD. Odds ratios were 1.5 (P = 0.035) in the original sample and 2.1 (P = 0.018) in the replication sample. Association fine mapping with 80 SNPs located within 50 kilobases of the marker SNP identified a 20 kilobase region of association containing exon 6 of PDE4D. In a second, family-based replication sample with a preponderance of females with low BMD, rs1498608 showed an opposite relationship with BMD at different sites (p = 0.00044-0.09). We also replicated the previously reported association of the Ser37Ala polymorphism in BMP2, known to interact biologically with PDE4D, with BMD. CONCLUSION: This study indicates that variants in the gene encoding PDE4D account for some of the genetic contribution to bone mineral density variation in humans. The contrasting results from different samples indicate that the effect may be context-dependent. PDE4 inhibitors have been shown to increase bone mass in normal and osteopenic mice, but up until now there have been no reports implicating any member of the PDE4 gene family in human osteoporosis. PMID- 15752432 TI - wFleaBase: the Daphnia genome database. AB - BACKGROUND: wFleaBase is a database with the necessary infrastructure to curate, archive and share genetic, molecular and functional genomic data and protocols for an emerging model organism, the microcrustacean Daphnia. Commonly known as the water-flea, Daphnia's ecological merit is unequaled among metazoans, largely because of its sentinel role within freshwater ecosystems and over 200 years of biological investigations. By consequence, the Daphnia Genomics Consortium (DGC) has launched an interdisciplinary research program to create the resources needed to study genes that affect ecological and evolutionary success in natural environments. DISCUSSION: These tools include the genome database wFleaBase, which currently contains functions to search and extract information from expressed sequenced tags, genome survey sequences and full genome sequencing projects. This new database is built primarily from core components of the Generic Model Organism Database project, and related bioinformatics tools. SUMMARY: Over the coming year, preliminary genetic maps and the nearly complete genomic sequence of Daphnia pulex will be integrated into wFleaBase, including gene predictions and ortholog assignments based on sequence similarities with eukaryote genes of known function. wFleaBase aims to serve a large ecological and evolutionary research community. Our challenge is to rapidly expand its content and to ultimately integrate genetic and functional genomic information with population-level responses to environmental challenges. URL: http://wfleabase.org/. PMID- 15752433 TI - Postage stamp multiple anterior capsulorhexisotomies in pediatric cataract surgery. AB - BACKGROUND: Capsule related complications are common following pediatric cataract surgery. We report a new technique of multiple anterior capsulorhexisotomies after lens aspiration and intraocular lens (IOL) implantation. METHODS: After performing automated lens aspiration, an IOL was implanted into the capsular bag. A bent 26 gauge needle was introduced through one side port and multiple small cuts were made in one half of the circumference of the anterior capsular rim by making a radial movement of the needle tip centripetally over the margin of the anterior capsular rim. The needle was again introduced through the other side port and multiple similar cuts were made in the other half thereby creating nearly 20 - 30 cuts at the margin of the anterior capsular rim. RESULTS: The mean size of the primary capsulorhexis was 4.33 +/- 0.20 mm. A uniform enlargement of the capsulorhexis could be performed in all the eyes without peripheral extension in any of the eyes. There was no damage to the posterior capsule and no scratch mark on the IOL. In one eye, the primary capsulorhexis was slightly eccentric, though it was covering the IOL optic all around. The rhexisotomies in this eye were limited to the capsular rim that was overlapping more on the IOL optic (sectoral anterior capsulorhexisotomies). CONCLUSION: The technique of postage stamp anterior capsulorhexisotomies is a feasible technique in pediatric cataracts. PMID- 15752435 TI - Managing febrile respiratory illnesses during a hypothetical SARS outbreak. AB - Since the World Health Organization declared the global outbreak of severe acute respiratory syndrome (SARS) contained in July 2003, new cases have periodically reemerged in Asia. This situation has placed hospitals and health officials worldwide on heightened alert. In a future outbreak, rapidly and accurately distinguishing SARS from other common febrile respiratory illnesses (FRIs) could be difficult. We constructed a decision-analysis model to identify the most efficient strategies for managing undifferentiated FRIs within a hypothetical SARS outbreak in New York City during the season of respiratory infections. If establishing reliable epidemiologic links were not possible, societal costs would exceed 2.0 billion US dollars per month. SARS testing with existing polymerase chain reaction assays would have harmful public health and economic consequences if SARS made up <0.1% of circulating FRIs. Increasing influenza vaccination rates among the general population before the onset of respiratory season would save both money and lives. PMID- 15752436 TI - Human disease from influenza A (H5N1), Thailand, 2004. AB - Influenza A (H5N1) is endemic in poultry across much of Southeast Asia, but limited information exists on the distinctive features of the few human cases. In Thailand, we instituted nationwide surveillance and tested respiratory specimens by polymerase chain reaction and viral isolation. From January 1 to March 31, 2004, we reviewed 610 reports and identified 12 confirmed and 21 suspected cases. All 12 confirmed case-patients resided in villages that experienced abnormal chicken deaths, 9 lived in households whose backyard chickens died, and 8 reported direct contact with dead chickens. Seven were children <14 years of age. Fever preceded dyspnea by a median of 5 days, and lymphopenia significantly predicted acute respiratory distress syndrome development and death. Among hundreds of thousands of potential human cases of influenza A (H5N1) in Asia, a history of direct contact with sick poultry, young age, pneumonia and lymphopenia, and progression to acute respiratory distress syndrome should prompt specific laboratory testing for H5 influenza. PMID- 15752437 TI - Lack of H5N1 avian influenza transmission to hospital employees, Hanoi, 2004. AB - To establish whether human-to-human transmission of influenza A H5N1 occurred in the healthcare setting in Vietnam, we conducted a cross-sectional seroprevalence survey among hospital employees exposed to 4 confirmed and 1 probable H5N1 case patients or their clinical specimens. Eighty-three (95.4%) of 87 eligible employees completed a questionnaire and provided a serum sample, which was tested for antibodies to influenza A H5N1. Ninety-five percent reported exposure to > or = 1 H5N1 case-patients; 59 (72.0%) reported symptoms, and 2 (2.4%) fulfilled the definition for a possible H5N1 secondary case-patient. No study participants had detectable antibodies to influenza A H5N1. The data suggest that the H5N1 viruses responsible for human cases in Vietnam in January 2004 are not readily transmitted from person to person. However, influenza viruses are genetically variable, and transmissibility is difficult to predict. Therefore, persons providing care for H5N1 patients should continue to take measures to protect themselves. PMID- 15752438 TI - Bacterial zoonoses and infective endocarditis, Algeria. AB - Blood culture-negative endocarditis is common in Algeria. We describe the etiology of infective endocarditis in this country. Samples from 110 cases in 108 patients were collected in Algiers. Blood cultures were performed in Algeria. Serologic and molecular analysis of valves was performed in France. Infective endocarditis was classified as definite in 77 cases and possible in 33. Causative agents were detected by blood cultures in 48 cases. All 62 blood culture-negative endocarditis cases were tested by serologic or molecular methods or both. Of these, 34 tested negative and 28 had an etiologic agent identified. A total of 18 infective endocarditis cases were caused by zoonotic and arthropodborne bacteria, including Bartonella quintana (14 cases), Brucella melitensis (2 cases), and Coxiella burnetii (2 cases). Our data underline the high prevalence of infective endocarditis caused by Bartonella quintana in northern Africa and the role of serologic and molecular tools for the diagnosis of blood culture-negative endocarditis. PMID- 15752439 TI - Novel flavivirus or new lineage of West Nile virus, central Europe. AB - A flavivirus (strain 97-103) was isolated from Culex pipens mosquitoes in 1997 following floods in South Moravia, Czech Republic. The strain exhibited close antigenic relationship to West Nile virus (WNV) prototype strain Eg-101 in a cross-neutralization test. In this study, mouse pathogenicity characteristics and the complete nucleotide and putative amino acid sequences of isolate 97-103, named Rabensburg virus (RabV) after a nearby Austrian city, were determined. RabV shares only 75%-77% nucleotide identity and 89%-90% amino acid identity with representative strains of WNV lineages 1 and 2. Another RabV strain (99-222) was isolated in the same location 2 years later; it showed >99% nucleotide identity to strain 97-103. Phylogenetic analyses of RabV, WNV strains, and other members of the Japanese encephalitis virus (JEV) complex clearly demonstrated that RabV is either a new (third) lineage of WNV or a novel flavivirus of the JEV group. PMID- 15752440 TI - Survey for bat lyssaviruses, Thailand. AB - Surveillance for lyssaviruses was conducted among bat populations in 8 provinces in Thailand. In 2002 and 2003, a total of 932 bats of 11 species were captured and released after serum collection. Lyssavirus infection was determined by conducting virus neutralization assays on bat serum samples. Of collected samples, 538 were either hemolysed or insufficient in volume, which left 394 suitable for analysis. These samples included the following: Pteropus lylei (n = 335), Eonycteris spelaea (n = 45), Hipposideros armiger (n = 13), and Rousettus leschennaulti (n = 1). No serum samples had evidence of neutralizing antibodies when tested against rabies virus. However, 16 samples had detectable neutralizing antibodies against Aravan virus, Khujand virus, Irkut virus, or Australian bat lyssavirus; all were specifically associated with fruit bats P. lylei (n = 15) and E. spelaea (n = 1). These results are consistent with the presence of naturally occurring viruses related to new putative lyssavirus genotypes. PMID- 15752441 TI - Spotted fever group and typhus group rickettsioses in humans, South Korea. AB - The presence of the nucleic acid of the spotted fever group (SPG) and typhus group (TG) rickettsiae was investigated in 200 serum specimens seropositive for SFG rickettsiae by multiplex-nested polymerase chain reaction with primers derived from the rickettsial outer membrane protein B gene. The DNA of SFG, TG, or both rickettsiae was amplified in the 24 serum specimens, and sequence analysis showed Rickettsia conorii, R. japonica, and R. felis in the specimens. R. conorii and R. typhi were found in 7 serum specimens, which indicated the possibility of dual infection in these patients. These findings suggest that several kinds of rickettsial diseases, including boutonneuse fever, rickettsialpox, R. felis infection, and Japanese spotted fever, as well as scrub typhus and murine typhus, are occurring in Korea. PMID- 15752443 TI - Sporadic cryptosporidiosis decline after membrane filtration of public water supplies, England, 1996-2002. AB - The incidence of sporadic cryptosporidiosis among 106,000 residents of 2 local government districts in northwest England before and after installation of membrane filtration of public water supplies was compared to that of 59,700 residents whose public water supplies remained unchanged. A national outbreak of foot and mouth disease in livestock during 2001 was associated with a decline in sporadic human cryptosporidiosis in all regions of the United Kingdom. In a Poisson regression model, membrane filtration was associated with an estimated 79% reduction (incidence ratio 0.207, 95% confidence intervals 0.099-0.431, p < 0.0001) after adjustment for the interval of the foot and mouth disease epidemic and the water source. Despite the confounding effect of that epidemic, membrane filtration of the public water supply was effective in reducing the risk for sporadic human Cryptosporidium infection in this population. PMID- 15752442 TI - Pneumocystis jirovecii in general population. AB - The possible presence of Pneumocystis among healthy adults was examined by detecting Pneumocystis jirovecii-specific DNA in prospectively obtained oropharyngeal wash samples from 50 persons without underlying lung disease or immunosuppression. Pneumocystis carriage, defined by detecting Pneumocystis DNA by nested polymerase chain reaction in 2 independent analyses plus successful mitochondrial large subunit ribosomal RNA typing by direct sequencing, was found in 20% of cases. All carriers were asymptomatic, anti-HIV negative, and had normal total lymphocyte and CD4+ cell counts. A second sample obtained in the 6 month follow-up was positive in 2 of 9 available carriers. Genotype analysis showed different polymorphisms; 85A/248C (40%) and 85C/248C (30%) were most frequently observed. This study provides the first evidence that P. jirovecii DNA can be frequently detected in the respiratory tract of immunocompetent adults, which agrees with the hypothesis that the general population could be a reservoir and source of this infection. PMID- 15752444 TI - Carbapenemase-producing Enterobacteriaceae, U.S. rivers. AB - Our study was initiated by previous isolation of 30 imipenem-resistant, gram negative rods from 7 of 16 U.S. rivers sampled from 1999 to 2001. Imipenem hydrolysis was detected in 22 of those isolates identified as Enterobacter asburiae. Random amplified polymorphism DNA analysis showed that these E. asburiae isolates were genetically indistinguishable. An identical clavulanic acid-inhibited beta-lactamase IMI-2 was identified from each isolate that shared 99% and 97% amino acid identity with the chromosome-encoded beta-lactamases IMI-1 and NmcA, respectively, from E. cloacae clinical isolates. The blaIMI-2 gene was located on a self-transferable 66-kb plasmid. Sequence analysis of a cloned 5.5 kb DNA fragment obtained from 1 of the imipenem-resistant E. asburiae isolates identified an upstream LysR-type regulator gene that explained inducibility of IMI-2 expression. beta-Lactamase IMI-2 is the first inducible and plasmid-encoded carbapenemase. Identification of clonally related E. asburiae isolates from distant rivers indicates an environmental and enterobacterial reservoir for carbapenemase genes. PMID- 15752445 TI - Rickettsial infection in animals and Brazilian spotted fever endemicity. AB - We compared the rickettsial infection status of Amblyomma cajennense ticks, humans, dogs, and horses in both Brazilian spotted fever (BSF)-endemic and nonendemic areas in the state of Sao Paulo, Brazil. Most of the horses and few dogs from BSF-endemic areas had serologic titers against Rickettsia rickettsii antigens. In contrast, no dogs or horses from BSF-nonendemic areas had serologic titers against R. rickettsii antigens, although they were continually exposed to A. cajennense ticks. All human serum samples and ticks from both areas were negative by serologic assay and polymerase chain reaction, respectively. Our results indicate that surveys of horse serum are a useful method of BSF surveillance in areas where humans are exposed to A. cajennense ticks. In addition, we successfully performed experimental infection of A. cajennense ticks with R. parkeri. PMID- 15752446 TI - Isolation of Waddlia malaysiensis, a novel intracellular bacterium, from fruit bat (Eonycteris spelaea). AB - An obligate intracellular bacterium was isolated from urine samples from 7 (3.5%) of 202 fruit bats (Eonycteris spelaea) in peninsular Malaysia. The bacterium produced large membrane-bound inclusions in human, simian, and rodent cell lines, including epithelial, fibroblastlike, and lymphoid cells. Thin-section electron microscopy showed reticulate bodies dividing by binary fission and elementary bodies in the inclusions; mitochondria surrounded the inclusions. The inclusions were positive for periodic acid-Schiff stain but could not be stained by fluorescein-labeled anti-Chlamydia trachomatis major outer membrane protein monoclonal antibody. The bacterium was resistant to penicillin and streptomycin (MICs > 256 mg/L) but susceptible to tetracycline (MIC = 0.25 mg/L) and chloramphenicol (MIC = 0.5 mg/L). Sequence analysis of the 16SrRNA gene indicated that it was most closely related to 2 isolates of Waddlia chondrophila (94% and 96% identity). The 16S and 23S rRNA gene signatures were only 91% identical. We propose this novel bacterium be called W. malaysiensis. PMID- 15752447 TI - Quarantine for SARS, Taiwan. AB - During the 2003 outbreak of severe acute respiratory syndrome (SARS) in Taiwan, >150,000 persons were quarantined, 24 of whom were later found to have laboratory confirmed SARS-coronavirus (SARS-CoV) infection. Since no evidence exists that SARS-CoV is infective before the onset of symptoms and the quarantined persons were exposed but not symptomatic, we thought the quarantine's effectiveness should be investigated. Using the Taiwan quarantine data, we found that the onset to-diagnosis time of previously quarantined confirmed case-patients was significantly shortened compared to that for those who had not been quarantined. Thus, quarantine for SARS in Taiwan screened potentially infective persons for swift diagnosis and hospitalization after onset, thereby indirectly reducing infections. Full-scale quarantine measures implemented on April 28 led to a significant improvement in onset-to-diagnosis time of all SARS patients, regardless of previous quarantine status. We discuss the temporal effects of quarantine measures and other interventions on detection and isolation as well as the potential usefulness of quarantine in faster identification of persons with SARS and in improving isolation measures. PMID- 15752448 TI - Wild animal mortality monitoring and human Ebola outbreaks, Gabon and Republic of Congo, 2001-2003. AB - All human Ebola virus outbreaks during 2001-2003 in the forest zone between Gabon and Republic of Congo resulted from handling infected wild animal carcasses. After the first outbreak, we created an Animal Mortality Monitoring Network in collaboration with the Gabonese and Congolese Ministries of Forestry and Environment and wildlife organizations (Wildlife Conservation Society and Programme de Conservation et Utilisation Rationnelle des Ecosystemes Forestiers en Afrique Centrale) to predict and possibly prevent human Ebola outbreaks. Since August 2001, 98 wild animal carcasses have been recovered by the network, including 65 great apes. Analysis of 21 carcasses found that 10 gorillas, 3 chimpanzees, and 1 duiker tested positive for Ebola virus. Wild animal outbreaks began before each of the 5 human Ebola outbreaks. Twice we alerted the health authorities to an imminent risk for human outbreaks, weeks before they occurred. PMID- 15752449 TI - Surveillance and control measures during smallpox outbreaks. AB - We reviewed historical data from 2 smallpox outbreaks in Liverpool and Edinburgh during the early and middle years of the 20th century to assess their contribution to developing modern strategies for response to a deliberate release of smallpox virus. Reports contemporaneous to these outbreaks provide detail on the effectiveness of public health interventions. In both outbreaks, extensive contact tracing, quarantine, and staged vaccination campaigns were initiated, and the outbreaks were controlled within 15 months and 3 months, respectively. In Edinburgh, the number of fatalities associated with vaccination exceeded the number of deaths from the disease. In Liverpool, ambulatory, vaccine-modified cases and misdiagnosis as chickenpox resulted in problems with outbreak control. The relatively slow spread of smallpox, as exemplified by the report from Liverpool, allowed for effective implementation of targeted intervention methods. Targeted surveillance and containment interventions have been successful in the past and should be explored as alternatives to mass vaccination. PMID- 15752450 TI - In vitro host-cell susceptibility to Usutu virus. AB - We investigated the susceptibility to Usutu virus (Flavivirus) of 13 permanent cell lines, 3 primary cell cultures, and chicken embryos. Vero, PK-15, and goose embryo fibroblast cells developed cytopathic effects; however, viral multiplication was detected in all mammalian cell types by immunohistochemical tests. Chicken embryo fibroblast cells and chicken embryos were resistant. PMID- 15752451 TI - Bat incidents at children's camps, New York State, 1998-2002. AB - From 1998 to 2002, a total of 299 bat incidents were reported at 109 children's camps in New York; 1,429 campers and staff were involved, and 461 persons received rabies treatment. In 53.8% of the incidents, the bat was captured and samples tested negative for rabies virus, which resulted in 61.3% of persons not receiving rabies treatment. PMID- 15752452 TI - West Nile virus in Morocco, 2003. AB - West Nile virus (WNV) reemerged in Morocco in September 2003, causing an equine outbreak. A WNV strain isolated from a brain biopsy was completely sequenced. On the basis of phylogenetic analyses, Moroccan WNV strains isolated during the 1996 and 2003 outbreaks were closely related to other strains responsible for equine outbreaks in the western Mediterranean basin. PMID- 15752453 TI - Diagnostic system for rapid and sensitive differential detection of pathogens. AB - Naturally emerging and deliberately released pathogens demand new detection strategies to allow early recognition and containment. We describe a diagnostic system for rapid, sensitive, multiplex discrimination of microbial gene sequences and report its application for detecting 22 respiratory pathogens in clinical samples. PMID- 15752454 TI - Comparing aberration detection methods with simulated data. AB - We compared aberration detection methods requiring historical data to those that require little background by using simulated data. Methods that require less historical data are as sensitive and specific as those that require 3-5 years of data. These simulations can determine which method produces appropriate sensitivity and specificity. PMID- 15752455 TI - Malaria epidemic and drug resistance, Djibouti. AB - Analysis of Plasmodium falciparum isolates collected before, during, and after a 1999 malaria epidemic in Djibouti shows that, despite a high prevalence of resistance to chloroquine, the epidemic cannot be attributed to a sudden increase in drug resistance of local parasite populations. PMID- 15752456 TI - Late recognition of SARS in nosocomial outbreak, Toronto. AB - Late recognition of severe acute respiratory syndrome (SARS) was associated with no known SARS contact, hospitalization before the nosocomial outbreak was recognized, symptom onset while hospitalized, wards with SARS clusters, and postoperative status. SARS is difficult to recognize in hospitalized patients with a variety of underlying conditions in the absence of epidemiologic links. PMID- 15752457 TI - Bacteremic typhoid fever in children in an urban slum, Bangladesh. AB - We confirmed a bacteremic typhoid fever incidence of 3.9 episodes/1,000 person years during fever surveillance in a Dhaka urban slum. The relative risk for preschool children compared with older persons was 8.9. Our regression model showed that these children were clinically ill, which suggests a role for preschool immunization. PMID- 15752458 TI - Molecular evidence of Pneumocystis transmission in pediatric transplant unit. AB - We describe an outbreak of Pneumocystis jirovecii pneumonia in a pediatric renal transplant unit, likely attributable to patient-to-patient transmission. Single strand conformation polymorphism molecular typing showed that 3 affected patients had acquired the same 2 strains of Pneumocystis, which suggests interhuman infection. An infant with mitochondriopathy was the probable index patient. PMID- 15752459 TI - Distinguishing levuglandins produced through the cyclooxygenase and isoprostane pathways. AB - The cyclooxygenase (COX) pathway generates enantiomerically pure levuglandin (LG) E(2) by a rearrangement of the prostaglandin (PG) endoperoxide PGH(2). The isoprostane pathway generates racemic LGE(2) together with stereoisomers, designated collectively as isoLGE(2), through free radical-induced lipid oxidation. Within seconds, both LGs and isoLGs are rapidly sequestered by protein adduction. In theory, the diastereomeric purity of LGE(2)-protein adduct-derived lysyl lactams can reveal the relative contributions of the COX and isoprostane pathways to LGE(2) stereoisomer production in vivo. Notably, however, the detection of LGE(2)-protein adducts does not provide a basis for inferring their formation through the isoprostane pathway in vivo unless the COX pathway can be rigorously excluded. In contrast, LGE(2)structural isomers, designated collectively as iso[n]LGE(2)s, are produced exclusively through the isoprostane pathway. Immunoassays that selectively recognize iso[n]LGE(2)-protein adducts are the only tools available to unambiguously detect and quantify the production of isolevuglandins in vivo through free radical-induced oxidation of arachidonates. PMID- 15752460 TI - A histological and NMR study of the melon of the striped dolphin (Stenella coeruleoalba). AB - The melon, the echolocation organ of the striped dolphin (Stenella coeruleoalba), was investigated by morphological and high-resolution (13)C nuclear magnetic resonance (NMR) methods, in order to characterize structure and composition gradient at the histological and molecular level. Morphological analysis showed that the lipidic components are organized in an adipose tissue; moreover, a clear muscular component was observed. Age-related structural differences also were noted. Furthermore, NMR yielded detailed information at a qualitative quantitative level on the lipid components. PMID- 15752462 TI - Free-radical fragmentation of galactocerebrosides: a MALDI-TOF mass spectrometry study. AB - Analysis of final products of radiation-induced transformations of galactocerebrosides (GalCer) in aqueous dispersions has been performed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and its combination of thin-layer chromatography (TLC). Ceramides were found to be the main products of GalCer gamma-radiolysis. From experimental results obtained in this study, as well as from the data available in the literature, an inference is made that the formation of ceramides occurs owing to fragmentation of radicals with an unpaired electron of the C2 atom of the carbohydrate moiety, formed from the starting compounds. PMID- 15752463 TI - FTIR spectroscopic studies of lipid dynamics in phytosphingosine ceramide models of the stratum corneum lipid matrix. AB - IR spectroscopic studies are reported for N-stearyl-D-erythro-phytosphingosine (Cer NP) and N-stearyl-2-hydroxy-D-erythro-phytosphingosine (Cer AP) in a hydrated model of the skin lipid barrier comprised of equimolar mixtures of each ceramide with cholesterol and d(35)-stearic acid. Examination of the methylene stretching, rocking and bending modes reveal some rotational freedom and hexagonal packing in both the ceramide and stearic acid chains. Analysis of the acid carbonyl stretch and the ceramide Amide I modes show both shift to higher frequencies, indicating weaker hydrogen bonding, in the mixed systems compared to the pure materials. For both systems, the fatty acid chain disordering temperatures are significantly increased from those of the pure acids. The observed behaviors of these phytosphingosine ceramide systems are fundamentally different from the previously reported analogous sphingosine ceramide systems. The implications of these observations for lipid organization in the stratum corneum are briefly discussed. PMID- 15752461 TI - Thermodynamic and structural characterization of amino acid-linked dialkyl lipids. AB - Using differential scanning calorimetry (DSC), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), we determined some thermodynamic and structural parameters for a series of amino acid-linked dialkyl lipids containing a glutamic acid-succinate headgroup and di-alkyl chains: C12, C14, C16 and C18 in CHES buffer, pH 10. Upon heating, DSC shows that the C12, C14 and annealed C16 lipids undergo a single transition which XRD shows is from a lamellar, chain ordered subgel phase to a fluid phase. This single transition splits into two transitions for C18, and FTIR shows that the upper main transition is predominantly the melting of the hydrocarbon chains whereas the lower transition involves changes in the headgroup ordering as well as changes in the lateral packing of the chains. For short incubation times at low temperature, the C16 lipid appears to behave like the C18 lipid, but appropriate annealing at low temperatures indicates that its true equilibrium behavior is like the shorter chain lipids. XRD shows that the C12 lipid readily converts into a highly ordered subgel phase upon cooling and suggests a model with untilted, interdigitated chains and an area of 77.2A(2)/4 chains, with a distorted orthorhombic unit subcell, a=9.0A, b=4.3A and beta=92.7 degrees . As the chain length n increases, subgel formation is slowed, but untilted, interdigitated chains prevail. PMID- 15752464 TI - Interaction of L-arginine with dihexadecylphosphate unilamellar liposomes: the effect of the lipid phase organization. AB - The interaction of L-arginine with unilamellar liposomes of dihexadecylphosphate sodium salt (DHP-Na) has been investigated using calorimetric, light scattering, fluorescence spectroscopy and zeta-potential techniques. Heating from room temperature, the bilayer exhibits a phase transition from a subgel (L(c)) to the gel (L(beta')) phase as well as a pre-transition (L(beta')-P(beta')), which is followed by the main lipid phase transition (P(beta')-L(alpha)). Direct studies of the interaction of L-arginine with the DHP-Na bilayers via isothermal titration calorimetry at 27 degrees C depict significant differences between samples in the L(c) and the L(beta') phases reflecting the effect of molecular organization of the lipids upon the interaction. While L-arginine has only a small impact upon the L(c) to L(beta') phase transition, it affects more significantly the transition temperature as well as the shape of the DSC peaks of the main lipid phase transition. Based on fluorescence and zeta-potential studies, the permeability of L-arginine through the liposomal membrane is higher within the temperature range of the main lipid phase transition. Encapsulated l arginine obstructs the formation of the subgel phase. PMID- 15752465 TI - Iron affects the structure of cell membrane molecular models. AB - The effects of Fe(3+) and Fe(2+) on molecular models of biomembranes were investigated. These consisted of bilayers of dimyristoylphosphatidylcholine (DMPC) and of dimyristoylphosphatidylethanolamine (DMPE), classes of phospholipids located in the outer and inner moieties of cell membranes, respectively. X-ray studies showed that very low concentrations of Fe(3+) affected DMPC organization and 10(-3)M induced a total loss of its multilamellar periodic stacking. Experiments carried out with Fe(2+) on DMPC showed weaker effects than those induced by Fe(3+) ions. Similar experiments were performed on DMPE bilayers. Fe(3+) from 10(-7)M up to 10(-4)M had practically no effect on DMPE structure. However, 10(-3)M Fe(3+) induced a deep perturbation of the multilamellar structure of DMPE. However, 10(-3)M Fe(2+) had no effect on DMPE organization practically. Differential scanning calorimetry measurements also revealed different effects of Fe(3+) and Fe(2+) on the phase transition and other thermal properties of the examined lipids. In conclusion, the results obtained indicate that iron ions interact with phospholipid bilayers perturbing their structures. These findings are consistent with the observation that iron ions change cell membrane fluidity and, therefore, affect its functions. PMID- 15752466 TI - Monitoring cholesterol organization in membranes at low concentrations utilizing the wavelength-selective fluorescence approach. AB - We previously showed using a fluorescent analogue of cholesterol (NBD cholesterol, or 25-[N-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-methyl]amino]-27 norcholesterol), that cholesterol may exhibit local organization at low concentrations in membranes by the formation of transbilayer tail-to-tail dimers of cholesterol (Rukmini, R., Rawat, S.S., Biswas, S.C., Chattopadhyay, A., 2001. Biophys. J. 81, 2122-2134). In this report, we have monitored the microenvironmental features of cholesterol monomers and dimers utilizing wavelength-selective fluorescence spectroscopy. Our results utilizing red edge excitation shift (REES) and wavelength-dependent change in fluorescence anisotropy show that the microenvironment around the NBD moieties in the dimer form is more rigid possibly due to steric constraints imposed by the dimer conformation. These results provide new information and are relevant in understanding the organization of cholesterol in membranes at low concentrations. PMID- 15752467 TI - Antitumoural properties of benzannelated seven-membered 5-fluorouracil derivatives and related open analogues. Molecular markers for apoptosis and cell cycle dysregulation. AB - Attention is increasingly being focussed on the cell cycle and apoptosis as potential targets for therapeutic intervention in cancer. We prepared a series of bioisosteric benzannelated seven-membered 5-FU O,N-acetals to test them against the MCF-7 human breast cancer cell line. Benzo-fused seven-membered O,O-acetals or their acyclic analogues led to the expected 5-FU O,N-acetals (or aminals), in addition to six- and 14-membered aminal structures and acyclic compounds. All the cyclic aminals provoked a G0/G1-phase cell cycle arrest, whereas Ftorafur, a known prodrug of 5-FU, and 1-[2-(2-hydroxymethyl-4-nitrophenoxy)-1-methoxyethyl] 5-fluorouracil (11) induced an S-phase cell cycle arrest. Although breast cancer is most often treated with conventional cytotoxic agents it has proved difficult to induce apoptosis in breast cancer cells, but improved clinical responses may be obtained by identifying therapies that are particularly effective in activating apoptosis. 1-(2,3-Dihydrobenzoxepin-2-yl)-5-fluorouracil (5) may be particularly useful in stimulating apoptosis in breast cancer. PMID- 15752468 TI - Rigid analogs of DMPP as probes for the nicotinic receptors. AB - Chemical manipulation of the nicotinic agonist DMPP, endowed with modest activity on the central receptors, definitely improved its affinity and pharmacokinetic properties. Although their pharmacophore is somehow different from that of classical nicotinic ligands, some DMPP derivatives show low nanomolar affinity for the central nicotinic receptors. Introduction of rigidity in the structure of DMPP and in that of its analogue 1-(3-pyridyl)piperazine, resulted in molecules with lower or null affinity for the central nicotinic receptors. This suggests that the frozen structures chosen either do not represent the bioactive conformation, or their volume is not compatible with the space available within the interaction site. PMID- 15752469 TI - Synthesis and in vitro pharmacological activity of oxypropanol analogs of labetalol. AB - New oxypropanol alpha- and beta-adrenoceptor blocking agents, analogs of labetalol, were synthesised and studied in vitro in left atria and aorta for alpha and beta activity. PMID- 15752471 TI - Synthesis of 2,3,5,6-tetrahydro-3H-imidazo[2,1-b] [1,3,5]benzotriazepines and their oxidative ring contraction into 1-(4,5-dihydro-1H-imidazol-2-yl)-1H benzimidazoles. AB - Syntheses of novel 5-aryl-2,3,5,6-tetrahydro-3H-imidazo[2,1-b] [1,3,5]benzotriazepine derivatives 3a-g were performed by reacting 2-(2 aminoarylimino)imidazolidines 1a-b with corresponding aryl aldehydes. The compounds 3 incorporating aminal group upon treatment with 2,3-dichloro-5,6 dicyano-1,2-benzoqinone (DDQ) underwent the oxidative ring contraction to give 1 (4,5-dihydro-1H-imidazol-2-yl)-2-aryl-benzimidazoles 4a-g. Reactions of the compounds 1a-c with carbonyldiimidazole (CDI) afforded novel 2,3,5,6-tetrahydro 1H-imidazo[2,1-b] [1,3,5]benzotriazepin-5-ones 5a-c which when heated in boiling methanol gave the corresponding 1-(4,5-dihydro-1H-imidazol-2-yl)-1,3-dihydro-2H benzimidazol-2-ones 6a-c. Radioligand binding studies using rat central imidazoline I2 receptors and alpha2-adrenoceptors demonstrated that benzimidazoles 4a-g display a low affinity (microM) for these receptors while benzimidazol-2-ones 6a-b elicited a moderate affinity for I2 receptor with Ki values of 490 and 220 nM, respectively. PMID- 15752470 TI - 1,5-Benzodiazepines XIV. Synthesis of new substituted 9H-bis-[1,2,4]triazolo[4,3 a:3',4'-d] [1,5]benzodiazepines and relate compounds endowed with in vitro cytotoxic properties. AB - A series of 11 new 9H-bis-[1,2,4]triazolo[4,3-a:3',4'-d] [1,5]benzodiazepine derivatives 8e-o was synthesized. Ten of these compounds (8e-m,o), along with four analogues (8a-d) (previously synthesized by us) were tested in vitro in order to evaluate their cytotoxic and anti-HIV-1 properties. In this connection other six original compounds, i.e., five 9-substituted compounds prepared starting from the 6,12-diphenylderivative 8c (compounds 10, 11, 12, 13a,b) and the bis-triazolone derivative 14, were synthesized and tested for the same purpose. While none of the 20 compounds tested exhibited any appreciable anti-HIV 1 activity, some of them exhibited interesting cytotoxic properties, the best results being shown by compounds 8c,d,k and 11 (CC(50) range=3-12 microM). Therefore, these four compounds were further evaluated for their antiproliferative activity against a panel of human tumor cell lines; actually, compounds 8d, 8k and 11 showed antiproliferative properties against either or both leukemia- and lymphoma-derived cell lines in the low micromolar range. PMID- 15752472 TI - Homopterocarpanes as bridged triarylethylene analogues: synthesis and antagonistic effects in human MCF-7 breast cancer cells. AB - A series of new compounds structurally derived from 6a,12a-dihydro-6H,7H-[1] benzopyran-[4,3-b]-benzopyran (homopterocarpane) was efficiently synthesized by reduction of the corresponding pyrilium salts obtained by treatment of selected flavanones and aldehydes with anhydrous HClO4. Cytotoxic effects on the human breast cancer cell line MCF-7 and antiestrogenic activity (only for compounds which resulted more active than tamoxifen (TAM)) on MCF-7 cells stimulated by 17beta-estradiol were evaluated. In vivo antiestrogenic activity and the relative binding affinity were also assessed. Some of the new compounds (4c, 4h, 4i and 4l) showed a biological activity in the micromolar range, and were more potent than TAM taken as the reference. PMID- 15752473 TI - Evaluation of the electrochemical behavior and analytical potentialities of a carbon paste electrode modified with a ruthenium (III) piperidinedithiocarbamate complex. AB - The preparation and electrochemical characterization of a carbon paste electrode modified with bis(N,N-piperidinedithiocarbamate)-mu-tris(N,N piperidinedithiocarbamate)diruthenium(III) complex, alpha-[Ru2(Pip)5]Cl are described. The best voltammetric response was obtained for a 10% (m/m) alpha [Ru2(Pip)5]Cl content in the paste, potassium acid phthalate solution pH 4.0 as supporting electrolyte and scan rate of 100 mV s-1. The analytical potentialities of the electrode have been evaluated using L-ascorbic acid (vitamin C) as a probe. A sensitive linear voltammetric response for L-ascorbic acid was obtained in the concentration range 4.50-113x10(-5) mol l-1 (7.92-200 mg l-1) with a slope of 1.12x10(4) microA mol-1, and a detection limit (3sigma/slope) of 7.00x10(-6) mol l-1 using cyclic voltammetry. The concentrations of L-ascorbic acid in pharmaceutical formulations (tablets and liquid) were determined using the modified electrode and compared with an iodimetric procedure with good agreement at the 95% confidence and relative error lower than 1%. PMID- 15752474 TI - Development and validation of RP-HPLC method for cetrimonium bromide and lidocaine determination. AB - The simple and rapid RP-HPLC method, for the simultaneous determination of lidocaine and cetrimonium bromide in the presence of pellet color corrigent, was developed. Separations were performed on a Beckman Ultrasphere ODS 4.6 mm x 15 cm, 5 microm particle column at 40 degrees C. The mobile phase consisted of water phase and acetonitrile (72:28 V/V), pH value of the mobile phase was adjusted to 2.0 with 85% ortophosphoric acid. Bisacodil was used as an internal standard. The flow rate was 1 ml/min and UV detection was performed at 208 nm. The proposed RP HPLC method was validated and all the parameters for the validation of the method are given. According to the obtained results, the developed method was found to be suitable and accurate for the determination of these drugs in commercial formulations. PMID- 15752475 TI - Determination of tramadol hydrochloride in ampoule dosage forms by using UV spectrophotometric and HPLC-DAD methods in methanol and water media. AB - Two newly developed simple and sensitive methods for determination of tramadol hydrochloride in ampoule dosage forms were described and validated. Measurements for spectrophotometric method were performed using UV-Vis Spectrophotometer in ranges of 200-400 nm. The solutions of standard and the samples were prepared in methanol and water media and the UV absorption spectrums of tramadol were monitored with maximum absorptions at 275 and 271 nm for both mediums, respectively. The standard calibration curves of tramadol were constructed by plotting absorbance vs. concentration in the concentration range with the final dilution of 10-100 microg ml-1. Reversed phase chromatography for HPLC method was conducted using a Phenomenex Bondclone C18 column with an isocratic mobile phase consisting of 25% acetonitrile in 75% 0.01 M phosphate buffer (pH 3). The effluent was monitored on a DAD detector at 218 nm. Linear response (r>0.99) was observed over the range of 0.5-40 microg ml-1 for methanol and water and run on six different occasions. The methods were applied successfully to pharmaceutical ampoule forms, but also for comparison in two different solvent media. Besides, it was completely validated and proven to be rugged. PMID- 15752476 TI - Preparation and in vitro evaluation of sustained release tablet formulations of diclofenac sodium. AB - The effects of formulation variables on the release profile of diclofenac sodium (DS) from hydroxypropylmethyl cellulose (HPMC) and chitosan matrix tablets were studied. DS tablets were prepared by wet granulation and direct compression methods and different ratios of HPMC and chitosan were used. Physical properties of the prepared tablets and targeted commercial sustained release (SR) tablet and the drug release were studied in tablets that were placed in 0.1 M HCl for 1 h and phosphate buffer solution was added to reach pH value of 7.5. In vitro studies showed that 20% HPMC contained SR formulation with direct (dry) compression method is the optimum formulation due to its better targeting profile in terms of release. This formulation also exhibited the best-fitted formulation into the zero order kinetics. The precision and accuracy of the analytical method were also checked. The repeatability and reproducibility of the method were also determined. PMID- 15752477 TI - A zucchini-peroxidase biosensor applied to dopamine determination. AB - A biosensor modified with peroxidase from crude extract of zucchini (Cucurbita pepo) was developed for the determination of dopamine in pharmaceutical samples. This enzyme catalyses the oxidation of dopamine to dopaminequinone, in presence of hydrogen peroxide, which the electrochemical reduction can be followed at a peak potential of -0.02 V. The recovery of dopamine from the samples ranged from 94.8% to 106% and a rectilinear analytical curve for dopamine concentration from 5.0 x 10(-4) to 3.0 x 10(-3) mol L-1 (r=0.9982) was obtained. The detection limit was 2.6x10(-5) mol L-1 and the relative standard deviation was less than 1.2% for 7.9 x 10(-4) mol L-1 dopamine in 0.1 mol L-1 phosphate buffer solution at pH 6.0 (n=10). PMID- 15752478 TI - Examining legal rules to protect children from injuries in recreational and sport activities. AB - Our jurisprudence assigns duties to persons to keep children safe. Under negligence law, a breach of duty causing an accident means the breaching party can be liable for damages inflicted on the injured person. Legislatures are considering new laws that reduce the damages that activity providers will need to pay to injured participants. Under some statutes, injured persons are precluded from maintaining lawsuits. In other cases, injured plaintiffs have a more stringent burden of proving liability. While activity providers may use insurance to pay for accident damages, for some injuries we might hold injured persons responsible. Four suggestions are offered as mechanisms to reduce tort litigation. PMID- 15752479 TI - Using persuasive messages to encourage voluntary hearing protection among coal miners. AB - INTRODUCTION: This longitudinal field study was designed to encourage Appalachian coal miners in West Virginia and Pennsylvania to engage in hearing-protection behaviors. METHOD: Participants were mailed postcards that featured either a positive, negative, or neutral message on the outside of the postcard and a message encouraging hearing protection behaviors on the inside. The first posttest measurement of the effectiveness of the persuasive messages was conducted about a week after the postcards were mailed. The delayed posttest measurement was conducted six weeks later. RESULTS: Responses from 307 coal miners revealed that the positive or neutral messages generated significantly more self-reported hearing protection behaviors than the negative message. Identical results were obtained in a delayed posttest assessment of miners' self reported hearing protection behaviors. The positive message was also more effective than either the neutral or negative message in preventing defensive mechanisms from emerging over time. IMPACT ON INDUSTRY: Positive and neutral messages were convincingly more successful than negative messages in facilitating self-reported hearing protection behaviors among coal miners. Similarly, the positive messages kept defensive processes at bay. PMID- 15752480 TI - Identifying best practices states in motorcycle rider education and licensing. AB - PROBLEM: After decreasing to a historic low in 1997, motorcycle crash-related fatalities are increasing. Although causes remain unclear, motorcycle rider education and licensing play key roles in reducing motorcycle crashes and injuries. Yet, little is known about what constitutes effective rider training and licensing. This study develops a model of best practices in motorcycle rider education and licensing and combines primary and secondary data to identify states that most closely adhere to this model. Evidence on the validity of the model is also examined. METHOD: States were rated along three areas of best practices: (a) program administration; (b) rider education; and (c) licensing based on 2001 data collected for a National Highway Traffic Safety Administration (NHTSA)-sponsored study. RESULTS: Results indicate wide variation in states' adherence to best practices; several states meet most, others very few. When the areas of best practices are considered separately, a state tends to behave similarly on all three. Initial evidence supports the validity of the model, with high best practices states having the lowest rates of motorcycle fatalities. IMPACT ON TRAFFIC SAFETY: As motorcycle-related crashes increase and state and federal support for rider education programs diminishes, it is critical that states identify deficiencies in their program and learn from successful states about efficient, cost-effective strategies for increasing best practices in motorcycle rider education and licensing. PMID- 15752481 TI - Probationary and non-probationary drivers' nighttime crashes in Western Australia, 1996-2000. AB - INTRODUCTION: This study was designed to explore the temporal aspects of crashes for probationary and non-probationary drivers. METHODS: Data from the West Australian Road Injury Database from 1996-2000 were used to calculate age-sex specific crash rates per 100,000 person-days and to plot proportions of fatal and hospital crashes by time for probationary and non-probationary drivers. The population attributable risk was used to estimate the potential number of lives saved by nighttime driving restriction in the probationary period. RESULTS: Probationary drivers were seven times more likely to crash than non-probationary drivers. While the highest number of crashes was in the daytime, probationary drivers had a higher proportion of fatal or hospitalization crashes at night than non-probationary drivers. CONCLUSION: Restrictions on driving at night could form part of graduated driver training. Even if some probationary drivers disobeyed the restriction, a substantial reduction in car occupant fatalities and hospitalizations could result. PMID- 15752482 TI - Safety rules and regulations on mine sites - the problem and a solution. AB - INTRODUCTION: Many accidents and incidents on mine sites have a causal factor in the rules and regulations that supposedly are in place to prevent the incident from occurring. The causes involve a lack of awareness or understanding, ignorance, or deliberate violations. The issue of mine rules, procedures, and regulations is a central focus of this paper, highlighted by this recent comment "very few people have accidents for which there is no procedure in place..." METHOD: An attitudinal survey was conducted at 33 mines throughout NSW, Queensland and international mine sites involving almost 500 mineworkers. The survey was in the form of a self-completing questionnaire, consisting of approximately 65 questions. It aimed to seek the opinions of the mining workforce on safety rules and regulations generally, as well as how they apply to their specific jobs on a mine site. The research also aimed to investigate: (a) the level of awareness and understanding of mine rules and procedures such as manager's rules and safe work procedures (SWPs); (b) the level of awareness and understanding of mine safety regulations and legislation; (c) the extent of communication of and commitment to rules and regulations; (d) the extent of compliance with rules and regulations; and (e) attitudes regarding errors, risk taking, and accidents and their interaction with rules and regulations. The sample consisted of a random selection of underground and open pit mines, extracting coal, metals, or industrial minerals. RESULTS: The insights provided by the mineworkers enabled a set of principles to be developed to guide mine management and regulators in the development of more effective rules and regulations. CONCLUSIONS AND IMPACT ON THE MINING INDUSTRY: (a) Management and regulators should not continue to produce more and more rules and regulations to cover every aspect of mining. (b) Detailed prescriptive regulations, detailed safe work procedures, and voluminous safety management plans will not "connect" with a miner. (c) Achieving more effective rules and regulations is not the only answer to a safer workplace. PMID- 15752483 TI - Physical limitations and musculoskeletal complaints associated with work in unusual or restricted postures: a literature review. AB - INTRODUCTION: The vast majority of ergonomics research has addressed the demands of work in standing or sitting postures, and understandably so. However, many workers (e.g., underground miners, aircraft baggage handlers, plumbers, agricultural workers, mechanics, and others) are often required to adopt postures such as kneeling, stooping, squatting, or lying down for significant periods of the workday. METHOD: A literature search was performed using the ISI Web of Science database (for years 1980-2004). Articles retrieved from this search were evaluated in terms of relevance to assessing physical capabilities of workers in these postures and/or the musculoskeletal epidemiology associated with these postures. RESULTS: Work in unusual and restricted postures was associated with significantly higher rates of musculoskeletal complaints compared to workers not adopting these postures in epidemiology studies (Odds Ratios ranging from 1.13 to 13). Some studies suggested a dose-response relationship, with longer exposures leading to increased musculoskeletal complaints. Physical strength and psychophysical lifting capacity vary significantly as unusual or restricted postures are adopted, with lower lifting capacities evident in the kneeling, squatting, and lying positions. CONCLUSIONS: Workers who adopt unusual or restricted postures appear to be at higher risk of musculoskeletal complaints and often exhibit reduced strength and lifting capacity. Research needs in this area include improved exposure assessment tools, studies of intervention effectiveness, adaptations of the body in response of work in unusual postures, and elucidation of relevant injury pathways. IMPACT ON INDUSTRY: Workers who adopt unusual or restricted postures in their work often experience higher musculoskeletal injury rates. If awkward postures cannot be eliminated in the workplace, jobs should be designed in accordance with the reduced strength and lifting capabilities observed in these postures. PMID- 15752484 TI - The prediction of safe lifting behavior: an application of the theory of planned behavior. AB - PROBLEM: Injuries resulting from lifting are costly, and create significant pain and discomfort. While engineering controls are the most effective means of reducing risks, most organizations continue to rely on manual lifting techniques. The problem, however, is that the use of safe-lifting techniques is inconsistent and managers have a difficult time motivating use of these techniques. Consequently, it is important to understand the factors driving safe-lifting behaviors. METHODS: This study used a survey to apply the theory of planned behavior (Ajzen., I., 1991. The theory of planned behavior. Organization Behavior and Human Processes, 50, 179-211) to safe-lifting among 136 materials management employees at a heavy manufacturing organization. Structural equation modeling and factor analysis were employed to analyze relationships among constructs. RESULTS: Results revealed that perceived behavioral control and intention were the strongest predictors of safe-lifting behavior. Subjective norms, to a lesser degree, were also important influences on intention. Attitudes did not surface as effective direct predictors of safe-lifting behavior, but did affect behavior and intent via mediating factors (subjective norms and perceived behavioral control). Finally, the theory of planned behavior was supported as an effective model explaining safe-lifting behavior, and had potential application for many other safety-related behaviors. IMPACT ON INDUSTRY: Results from this study emphasize the importance of perceived behavioral control as a factor associated with safety related behavior. PMID- 15752485 TI - Freight transport and non-driving work duties as predictors of falling asleep at the wheel in urban areas of Crete. AB - BACKGROUND: This study investigated the impact of subjective reports of drowsy driving and non-driving duties on the falling asleep responses and road crash involvement of professional drivers in Crete. An attempt was also made to elucidate other driving parameters, such as freight transportation, which could be potential predictors of risky driving, after controlling for lifestyle patterns. METHOD: A sample of 317 professional drivers was studied through personal interviews. The interview questionnaire included items about sleep and fatigue as contributing factors to falling asleep probability and crash risk. In addition, the drivers reported the type of freight they carried in their last trip, as well as practices such as smoking and alcohol consumption. RESULTS: The first logistic regression analysis showed that the most significant predictors of falling asleep at the wheel were transportation of fruits/vegetables and livestock, non-driving hours of work, insufficient hours of sleep, and smoking. The second logistic regression analysis revealed all the previous items as powerful factors of crash probability, including the transportation of express freight and freezer. IMPACT: The findings of the current study are discussed as they pertain to directions for future studies and for the development of fatigue countermeasures. PMID- 15752487 TI - Identifying crash propensity using specific traffic speed conditions. AB - INTRODUCTION: In spite of recent advances in traffic surveillance technology and ever-growing concern over traffic safety, there have been very few research efforts establishing links between real-time traffic flow parameters and crash occurrence. This study aims at identifying patterns in the freeway loop detector data that potentially precede traffic crashes. METHOD: The proposed solution essentially involves classification of traffic speed patterns emerging from the loop detector data. Historical crash and loop detector data from the Interstate-4 corridor in the Orlando metropolitan area were used for this study. Traffic speed data from sensors embedded in the pavement (i.e., loop detector stations) to measure characteristics of the traffic flow were collected for both crash and non crash conditions. Bayesian classifier based methodology, probabilistic neural network (PNN), was then used to classify these data as belonging to either crashes or non-crashes. PNN is a neural network implementation of well-known Bayesian-Parzen classifier. With its superb mathematical credentials, the PNN trains much faster than multilayer feed forward networks. The inputs to final classification model, selected from various candidate models, were logarithms of the coefficient of variation in speed obtained from three stations, namely, station of the crash (i.e., station nearest to the crash location) and two stations immediately preceding it in the upstream direction (measured in 5 minute time slices of 10-15 minutes prior to the crash time). RESULTS: The results showed that at least 70% of the crashes on the evaluation dataset could be identified using the classifiers developed in this paper. PMID- 15752486 TI - The relationship between protective and risk characteristics of acting and experienced workload, and musculoskeletal disorder cases among nurses. AB - PROBLEMS: Limited research is available on the acting (work characteristics) and experienced (perceived stress) workload of nurses. The relationship between risk and protective characteristics of work-related factors and the prevalence of musculoskletal symptoms in different body regions is also unclear. METHODS: The study was a cross-sectional design with 97 female registered nurses working in a hospital setting. Two surveys were used to document the workload exposure of the nurses. One survey consisted of 148 items aimed to measure the acting workload variables from the environment; the other survey included 33 items that were aimed to measure the nurses' experienced workload. The musculoskeletal outcomes were documented with a modified version of the Nordic Musculoskeletal Symptom Survey. RESULTS: Factor analyses revealed three factors that accounted for 56% of the total variance. Factor 1 (i.e., integrated experienced energy replenishment/expenditure) represented the psychological effects of work characteristics, effort, perceived risk, and performance. Factor 2 (i.e., acting energy replenishment/expenditure) consisted of non-physical variables of the work characteristics, while Factor 3 (i.e., acting energy expenditure) included both acting and experienced workload. Logistic regression analyses indicated that Factor 3 was significantly associated with the musculoskeletal symptoms of lower and upper back, hands/wrists, and knees/lower legs (odds ratios > 1.0). Factor 2 was significantly associated with the musculoskeletal symptoms of the upper back and knees/lower legs (odds ratios < 1.0). SUMMARY: Both the acting and experienced workloads exhibited associations with musculoskeletal outcomes in the lower back, upper back, hands/wrists, and knees/lower legs in terms of risk and protective effects. PMID- 15752488 TI - Photolysis of polychlorinated dibenzo-p-dioxins and dibenzofurans dissolved in vegetable oils: influence of oil quality. AB - Sunlight or ultraviolet light irradiation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in the presence of vegetable oil offers a potential method for the cleanup of contaminated soil. In this study, the effects of different types of vegetable oils on the photochemical degradation of 1,2,3,4,6,7,8-heptachlorodibenzofuran and heptachlorodibenzo-p-dioxin (1,2,3,4,6,7,8-HpCDF/HpCDD) were investigated in the laboratory. Using a blacklight lamp as a source of ultraviolet light, 93-100% of 1,2,3,4,6,7,8-HpCDF degraded in 60 min in rapeseed oil, extra virgin olive oil and olive oil. Less degradation occurred in palm oil (59%), toluene (39%) and hexane (20%). The better degradation in vegetable oils in comparison with organic solvents was attributed to the photooxidation of lipids producing hydrogen for PCDD/F dechlorination. In addition to the hydrogen donor capacity, permeability of ultraviolet light was involved in the differences between vegetable oils. alpha Tocopherol and chlorophyll did not influence the performance of oil at concentrations normally present in vegetable oils, whereas beta-carotene had an inhibitory effect on the degradation of 1,2,3,4,6,7,8-HpCDF. Up to 28% of the degradation products of 1,2,3,4,6,7,8-HpCDF were formed via the dechlorination pathway. Products included both toxic (2,3,7,8-chlorinated) and non-toxic PCDD/Fs, the toxic PCDD/Fs being more stable. Irradiation of 1,2,3,4,6,7,8-HpCDD yielded only non-toxic dechlorination products. Polychlorinated hydroxybiphenyls (OH-PCBs), polychlorinated dihydroxybiphenyls (DOH-PCBs) and polychlorinated hydroxydiphenylethers (OH-PCDEs) containing one to seven chlorine atoms were not detected in irradiated HpCDF/HpCDD samples. PMID- 15752489 TI - Air quality impacts of speed-restriction zones for road traffic. AB - This study investigates the air quality impacts of six 20 mph zones implemented in the North West of England. A monitoring study has been undertaken to measure the concentrations of nitrogen dioxide (NO2) and benzene in ambient air both before and after the implementation of the zones. An emissions estimation and simple dispersion modelling study has also been undertaken using traffic survey data collected at the zones at corresponding periods. The ambient air quality measurements and the modelling predictions did not show any significant impacts after implementation of the 20 mph zones. PMID- 15752490 TI - Quantifying and reducing uncertainty in life cycle assessment using the Bayesian Monte Carlo method. AB - The traditional life cycle assessment (LCA) does not perform quantitative uncertainty analysis. However, without characterizing the associated uncertainty, the reliability of assessment results cannot be understood or ascertained. In this study, the Bayesian method, in combination with the Monte Carlo technique, is used to quantify and update the uncertainty in LCA results. A case study of applying the method to comparison of alternative waste treatment options in terms of global warming potential due to greenhouse gas emissions is presented. In the case study, the prior distributions of the parameters used for estimating emission inventory and environmental impact in LCA were based on the expert judgment from the intergovernmental panel on climate change (IPCC) guideline and were subsequently updated using the likelihood distributions resulting from both national statistic and site-specific data. The posterior uncertainty distribution of the LCA results was generated using Monte Carlo simulations with posterior parameter probability distributions. The results indicated that the incorporation of quantitative uncertainty analysis into LCA revealed more information than the deterministic LCA method, and the resulting decision may thus be different. In addition, in combination with the Monte Carlo simulation, calculations of correlation coefficients facilitated the identification of important parameters that had major influence to LCA results. Finally, by using national statistic data and site-specific information to update the prior uncertainty distribution, the resultant uncertainty associated with the LCA results could be reduced. A better informed decision can therefore be made based on the clearer and more complete comparison of options. PMID- 15752491 TI - Iodine-129 in soils from Northern Ukraine and the retrospective dosimetry of the iodine-131 exposure after the Chernobyl accident. AB - Forty-eight soil profiles down to a depth of 40 cm were taken in Russia and Ukraine in 1995 and 1997, respectively, in order to investigate the feasibility of retrospective dosimetry of the 131I exposure after the Chernobyl accident via the long-lived 129I. The sampling sites covered areas almost not affected by fallout from the Chernobyl accident such as Moscow/Russia and the Zhitomir district in Ukraine as well as the highly contaminated Korosten and Narodici districts in Ukraine. 129I was analyzed by radiochemical neutron activation analysis (RNAA) and accelerator mass spectrometry (AMS). 127I was measured for some profiles by RNAA or ion chromatography (IC). The results for 127I demonstrated large differences in the capabilities of the soils to store iodine over long time spans. The depth profiles of 129I and of 137Cs showed large differences in the migration behavior between the two nuclides but also for each nuclide among the different sampling sites. Though it cannot be quantified how much 129I and 137Cs was lost out of the soil columns into deeper depths, the inventories in the columns were taken as proxies for the total inventories. For 129I, these inventories were at least three orders of magnitude higher than a pre nuclear value of 0.084+/-0.017 mBq m(-2) derived from a soil profile taken in 1939 in Lutovinovo/Russia. From the samples from Moscow and Zhitomir, a pre Chernobyl 129I inventory of (44+/-24) mBq m(-2) was determined, limiting the feasibility of 129I retrospective dosimetry to areas where the 129I inventories exceed 100 mBq m(-2). Higher average 129I inventories in the Korosten and Narodici districts of 130 and 848 mBq m(-2), respectively, allowed determination of the 129I fallout due to the Chernobyl accident. Based on the total 129I inventories and on literature data for the atomic ratio of 129I/131I=13.6+/-2.8 for the Chernobyl emissions and on aggregated dose coefficients for 131I, the thyroid exposure due to 131I after the Chernobyl accident was estimated for the inhabitants of four villages in the Korosten and of three villages in the Narodici districts. The limitations and uncertainties of the 129I retrospective dosimetry are discussed. PMID- 15752492 TI - Chemical water quality in Thailand and its impacts on the drinking water production in Thailand. AB - In Thailand, surface water and groundwater are the main water sources for tap water and drinking water production. Thirty-six different samples from surface waters from Chao Praya and Mae Klong rivers, tap waters, bottled drinking waters, groundwaters and commercial ice cubes from around the Bangkok area were collected. Water samples were also taken from two waterworks in the Chonburi province. The extensive survey showed that, overall in all water samples investigated, there was only a minor pollution which could be traced back to the analyses performed including amongst others total organic carbon, inorganics and heavy metals, pesticides, organochlorine compounds, volatile organic compounds, surfactants, pharmaceuticals and disinfection by-products. However, whenever organic micropollutants could be detected in surface water, such as, e.g. the herbicide atrazine, they were also present in the tap water produced thereof proving that the present treatment steps are not sufficient for removal of such pollutants. The concentration of disinfection by-products was higher in tap water produced from Chao Praya river than from Mae Klong river. Disinfection by products were also found in bottled drinking water. Commercial ice cubes contained anionic surfactants and their metabolites at elevated concentrations. The data of this study constitute the first set of homogenous data for the chemical water quality and also aid development of new water quality criteria in Thailand. PMID- 15752493 TI - Atmospheric polycyclic aromatic hydrocarbons: size distribution, estimation of their risk and their depositions to the human respiratory tract. AB - Polycyclic aromatic hydrocarbons (PAHs) in the atmosphere may be one of the causes of lung cancer. However, in many countries including Japan, only benzo[a]pyrene is regulated, though more toxic PAHs than benzo[a]pyrene have been reported. Therefore, it is necessary to characterize the risk of atmospheric PAHs and to find major contributing compounds that can act as suitable indicators for estimating and monitoring the risk of PAHs as a whole. The primary objectives of this study are to investigate the risk of 22 combined atmospheric PAHs and to find major contributing compounds that can act as adequate indicators for estimating the risk. The average lifetime cancer risk for all 22 PAHs in winter was 1.7 x 10(-4) in the Yokohama residential area. Although benzo[a]pyrene has been considered to be a useful indicator in many countries, the present study also showed that benzo[a]pyrene contributed only ca. 40% of the total cancer risk for all 22 PAHs, as calculated by unit risk. Six PAHs (dibenzo[a,h]anthracene, benzo[b]fluoranthene, indeno[1,2,3-cd]perylene, dibenzo[a,h]pyrene, dibenzo[a,e]pyrene, and benzo[a]pyrene) contributed 93% of the overall risk. Therefore, these six PAHs, including the high molecular ones, would be better indicators of the risk than benzo[a]pyrene alone. The secondary objective of this study is to reveal the PAHs that are major contributors to the deposition that contribute to the total deposition of PAHs in the human respiratory tract. The results revealed that the same six PAHs identified using unit risk evaluation were large contributors in all five parts of the respiratory tracts in adult males in spite of deposition process. It was concluded that the reason for this was that these six PAHs had the same size distributions, of which the 0.43-1.1 microm fraction was the most largest, about 57-70% of the whole concentration. PMID- 15752494 TI - Biosolids compost amendment for reducing soil lead hazards: a pilot study of Orgro amendment and grass seeding in urban yards. AB - In situ inactivation of soil Pb is an alternative to soil removal and replacement that has been demonstrated in recent years at industrial sites with hazardous soil Pb concentrations. Most children exposed to elevated soil Pb, however, reside in urban areas, and no government programs exist to remediate such soils unless an industrial source caused the contamination. Modern regulated biosolids composts have low Pb concentrations and low bioaccessible Pb fractions and can improve grass growth on urban soils. High Fe and P biosolids composts can reduce the bioavailability and bioaccessibility of soil Pb and can aid in establishing vegetation that would reduce soil transfer into homes. For these reasons, we conducted a field test of their use to reduce Pb bioaccessibility in urban soils in Baltimore, MD USA. We chose biosolids compost for its expected reduction in the bioaccessible Pb fraction of urban soils, ease of use by urban residents, and ability to beautify urban areas. Nine urban yards with mean soil Pb concentrations >800 mg Pb kg(-1) were selected and sampled at several distances from the house foundation before soil treatment. The soils were rototilled to 20 cm depth to prepare the sites, and resampled. The yards were then amended with 6 8 cm depth of Orgro biosolids compost (110-180 dry t/ha) rich in Fe and P, mixed well by rototilling, and resampled. Kentucky bluegrass (Poa pratensis) was seeded and became well established. Soils were resampled 1 year later. At each sampling time, total soil Pb was measured using a modified U.S. EPA nitric acid hotplate digestion method (SW 846 Method 3050) and bioaccessible Pb fraction was measured using the Solubility/Bioaccesibility Research Consortium standard operating procedure with modifications, including the use of glycine-buffered HCl at pH 2.2. Samples of untreated soils were collected from each yard and mixed well to serve as controls for the Pb bioaccessibility of field treated soils over time independent of positional variance within yards. At 1-year post-treatment, grass cover was healthy and reductions in bioaccessible Pb concentrations compared to pre-tillage were 64% (from 1655 to 595 mg kg(-1)) and 67% (from 1381 to 453 mg kg(-1)) at the sampling lines closest to the houses. Little or no reduction in bioaccessible Pb concentration was observed at sampling lines more remote from the house that also had the lowest bioaccessible Pb concentrations at pre-tillage (620 and 436 mg kg(-1), respectively). For the control soils, changes over time in total Pb and bioaccessible Pb concentrations and the bioaccessible Pb fraction were insignificant. This study confirms the viability of in situ remediation of soils in urban areas where children are at risk of high Pb exposure from lead in paint, dust and soil. PMID- 15752495 TI - Reproductive status in female perch (Perca fluviatilis) outside a sewage treatment plant processing leachate from a refuse dump. AB - Reproductive failure and endocrine disruption in female perch (Perca fluviatilis) and female brook trout (Salvelinus fontinalis), naturally exposed to toxic leachate from a municipal refuse dump, has raised concerns about the reproductive status in female fish outside the sewage treatment plant (STP), which now treats the same leachate. Female perch from the receiving water, Lake Siljan, were therefore sampled in a gradient from the STP in Fornby and Osterviken and compared with reference females from Isunda or Solleron. A high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) technique enabled the analysis of circulating steroids in blood plasma: the sex steroids progesterone (P), 17alpha-hydroxyprogesterone (17alpha-OHP), androstenedione (A), testosterone (T), estrone (E1), and 17beta-oestradiol (E2), the maturation inducing hormone (MIH) 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta P), and the glucocorticoids 11-deoxycortisol (11-DC) and cortisol (C). Body and organ measurements, ratio of sexually mature (SM) females, and frequencies of body lesions, were also recorded. The percentages of SM females found in Fornby on Oct 23 (83.8%) and in Osterviken on May 14 (54.0%) were lower compared with Isunda (100%). The lack of differences in gonadosomatic index (GSI) and liver somatic index (LSI) indicated a normal vitellogenesis in SM females from Fornby and Osterviken. No differences in circulating levels of steroids were observed between the sites, except at the time of spawning, when elevated P and 17alpha OHP levels in females from Osterviken indicated a steroidogenic shift from the synthesis of sex steroids towards the production of the MIH 17alpha,20beta-P. The significant peak-levels of both 17alpha,20beta-P and 11-DC in female perch from Osterviken at the same time suggest that both are involved in controlling final oocyte maturation in perch. The elevated levels of androgens (A and T) in females caught on the same occasion in Fornby implied that these females were at an earlier stage of oocyte maturation, prior to the steroidogenic shift towards MIH production. This study confirms the results of an earlier study revealing low numbers of SM female perch outside the STP in Fornby. The lack of associated effects on organ indexes and circulating levels of steroids does not indicate exposure to endocrine disrupting substances (EDSs) from the treated sewage. The consistently high numbers of SM females in the northern part of Siljan suggest, however, that natural factors are not responsible for the reproductive failures in the southern part of the lake. PMID- 15752496 TI - Accumulation of mercury in the tissues of the common octopus Octopus vulgaris (L.) in two localities on the Portuguese coast. AB - Mercury concentrations were measured in tissues of 12 individuals of Octopus vulgaris, captured by the commercial fishery at two points along the Portuguese coast, Viana do Castelo and Cascais, in spring 2002. Concentrations were determined in six tissues (digestive gland, branchial hearts, gills, mantle, arms, and gonads). Correlations between mercury concentrations in different tissues were examined as were correlations between mercury levels and total length, mantle length, weight, gonadosomatic index, digestive gland index, and state of maturation. Differences between sexes and localities were analysed. The concentration of mercury in the digestive gland (Viana, 0.58+/-0.08, and Cascais, 3.43+/-2.57 mg/kg dry weight) was higher than in the other tissues, and values were generally similar to those recorded in previous studies on octopods. Arm muscle contained most of the mercury with 56% of the total body burden followed by the digestive gland with 31%, mantle with 11%, gills with 0.8%, gonad with 0.5%, and branchial heart with 0.2%. In all tissues, mercury concentrations were slightly higher in samples from Cascais than in Viana do Castelo, which is consistent with higher concentrations recorded in seawater at Cascais. Levels of mercury determined in octopus were within the range of values legally defined as safe for human consumption. PMID- 15752497 TI - Chemical and ecotoxicological assessment of polycyclic aromatic hydrocarbon- contaminated sediments of the Niger Delta, Southern Nigeria. AB - The extent of environmental contamination and sources of polycyclic aromatic hydrocarbons (PAHs) compounds to sediments of the Niger Delta, Nigeria were assessed using combined chemical analysis and toxicity bioassay techniques. Concentrations of two- to six-ring PAHs of molecular mass 128-278 and toxicity to Vibrio fischeri and Lemna minor are considered in this investigation. Levels of the sum of the 16 USEPA priority pollutant PAHs varied from 20.7 to 72.1 ng/g dry weight. Sediment PAH levels were highest in samples collected from Delta Steel located at the outskirts of Warri, and Quality control centre, Ughelli West; with total PAH concentrations of 72.1 and 67.5 ng/g dry weight, respectively. The overall levels of PAHs in this study are low compared to other regions and reveal moderate PAHs pollution in the sediments of the Niger Delta. Two- and three-ring aromatic hydrocarbons predominated in almost all the sediments, which indicate a petrogenic origin. The sediment total PAH (PAHtot) concentration, normalized to organic carbon content (OC), ranged from 120.2 to 1.99 ng PAHtot/mg OC; and showed distinctively that the sedimentary organic matter of the sample from Delta Steel is highly contaminated with PAHs, and had a value of 120.2 ng PAHtot/mg OC. The toxicity bioassays indicated that the sample collected from Warri Refinery Area (SDWRR) was the most toxic to V. fischeri, with an EC50 value of 0.45 mg sediment equiv./mL test medium; and samples from Ogunu (SDOGN) and Warri Refinery area (SDWRR) showed high toxicity to L. minor, with percent inhibitions of 42.6% and 33.67%, respectively, after 7 days of exposure. The total PAH concentrations showed no correlation with toxicity bioassays, and thereby implied that chemical analysis of PAHs cannot be an indicator of sediment toxicity. PMID- 15752498 TI - Geochemical fingerprints and controls in the sediments of an urban river: River Manzanares, Madrid (Spain). AB - The geochemical fingerprint of sediment retrieved from the banks of the River Manzanares as it passes through the City of Madrid is presented here. The river collects the effluent water from several Waste Water Treatment (WWT) plants in and around the city, such that, at low flows, up to 60% of the flow has been treated. A total of 18 bank-sediment cores were collected along the course of the river, down to its confluence with the Jarama river, to the south-east of Madrid. Trace and major elements in each sample were extracted following a double protocol: (a) "Total" digestion with HNO3, HClO4 and HF; (b) "Weak" digestion with sodium acetate buffered to pH=5 with acetic acid, under constant stirring. The digests thus obtained were subsequently analysed by ICP-AES, except for Hg which was extracted with aqua regia and sodium chloride-hydroxylamine sulfate, and analysed by Cold Vapour-AAS. X-ray diffraction was additionally employed to determine the mineralogical composition of the samples. Uni- and multivariate analyses of the chemical data reveal the influence of Madrid on the geochemistry of Manzanares' sediments, clearly manifested by a marked increase in the concentration of typically "urban" elements Ag, Cr, Cu, Pb and Zn, downstream of the intersection of the river with the city's perimeter. The highest concentrations of these elements appear to be associated with illegal or accidental dumping of waste materials, and with the uncontrolled incorporation of untreated urban runoff to the river. The natural matrix of the sediment is characterised by fairly constant concentrations of Ce, La and Y, whereas changes in the lithology intersected by the river cause corresponding variations in Ca-Mg and Al-Na contents. In the final stretch of the river, the presence of carbonate materials seems to exert a strong geochemical control on the amount of Zn and, to a lesser extent, Cu immobilised in the sediments. This fact suggests that a variable but significant proportion of both elements may be susceptible to reincorporation in the aqueous phase under realistic environmental conditions. PMID- 15752499 TI - Sources and timing of anthropogenic pollution in the Ensenada de San Simon (inner Ria de Vigo), Galicia, NW Spain: an application of mixture-modelling and nonlinear optimization to recent sedimentation. AB - The Ensenada de San Simon is the inner part of the Ria de Vigo, one of the major mesotidal rias of the Galician coast, NW Spain. The geochemistry of its bottom sediments can be accounted for in terms of both natural and anthropogenic sources. Mixture-modelling enables much of the Cr, Ni, V, Cu, Pb and Zn concentrations of the bottom and subaqueous sediments to be explained by sediment input from the river systems and faecal matter from manmade mussel rafts. The compositions and relative contributions of additional, unknown, sources of anomalous heavy-metal concentrations are quantified using constrained nonlinear optimization. The pattern of metal enrichment is attributed to: material carried in solution and suspension in marine water entering the Ensenada from the polluted industrial areas of the adjacent Ria de Vigo; wind-borne urban dusts and/or vehicular emissions from the surrounding network of roads and a motorway road-bridge over the Estrecho de Rande; industrial and agricultural pollution from the R. Redondela; and waste from a former ceramics factory near the mouth of the combined R. Oitaben and R. Verdugo. Using (137)Cs dating, it is suggested that heavy metal build-up in the sediments since the late 1970s followed development of inshore fisheries and introduction of the mussel rafts (ca. 1960) and increasing industrialisation. PMID- 15752500 TI - Spatial and vertical distribution of polycyclic aromatic hydrocarbons in mangrove sediments. AB - The distribution and historical changes of polycyclic aromatic hydrocarbons (PAHs) contamination in mangrove sediments in Hong Kong SAR were investigated. Surface sediments (2-3 cm) collected from four mangrove swamps exhibited significant spatial variations in concentrations of total PAH (with SigmaPAHs ranging from 56 to 3758 ng g(-1) dry wt), as well as the composition of 16 USEPA priority PAH compounds. Within a small swamp with an area of 0.68 ha, the total PAH concentrations also differed from sampling site to site, indicating that the PAH contamination is localized and confined to a very small area within the same swamp. Discharges from municipal and industrial wastewater, urban runoff, oil leakage from boats and ships, and accidental oil spill are possible sources of the PAH contamination. The sediment depth profiles reveal that the surface sediment layer (0-5 cm) had lower total PAH concentrations than that in the bottom layer (15-20 cm), and PAH composition also changed with the sediment layers. Based on the estimated annual sediment deposition rate in Hong Kong SAR of around 0.4-0.5 cm, the present findings suggest that the PAH contamination was most serious between 1958 and 1979 but started to decline thereafter. Such decline was probably due to changes in petroleum usage in urban areas and a better control of wastewater discharges from 1980 onwards in this region. PMID- 15752501 TI - Source apportionment of fine and coarse atmospheric particles in Auckland, New Zealand. AB - Atmospheric fine particles (with a diameter less than 2.5 microm) and coarse particles (with a diameter between 2.5 and 10 microm) were collected simultaneously at an urban background site in Auckland, New Zealand, in the summer and winter of 2003. Major water-soluble ions, black carbon (BC), and some elements in the samples were analysed, and the data compiled into two sets. Selected trace gases (NH3, HONO, HNO3, and SO2) were also collected at the same time. Application of positive matrix factorisation to the particle data resolved five sources for the fine particles and five for the coarse particles. Weighted multiple linear regression was used to investigate the mass contributions of the sources. It was found that although natural sources had significant contributions to the coarse particles, anthropogenic sources dominated the contributions to the fine particles, particularly in winter. Significant seasonal changes of the sources were observed. Overall, sea salt and wind-blown soil had significantly stronger contributions in the summer; in contrast, the winter contribution of vehicle emissions, road dust, and industry and incineration sources was significantly higher. These seasonal changes may be explained by variations in meteorological conditions and atmospheric chemistry. The nonparametric correlations between the atmospheric concentrations of the trace gases and the mass contributions of the resolved sources demonstrate that these sources are realistic, providing a useful supplementary approach for the assessment of receptor modelling. PMID- 15752502 TI - Vertical distributions of 239+240Pu activities and 240Pu/239Pu atom ratios in sediment cores: implications for the sources of Pu in the Japan Sea. AB - The vertical distributions of 239+240Pu-specific activities and 240Pu/239Pu atom ratios in sediment cores of the Japan Sea were investigated. For comparison, the 239+240Pu-specific activities and 240Pu/239Pu atom ratios in the surface layer of bottom sediments in the Sea of Okhotsk were also determined. The 239+240Pu specific activities in the surface layer sediments varied from 0.43 to 2.65 mBq/g in the Japan Sea. The lowest value was seen in the eastern Japan Basin, while the highest value was found in a sample collected from a coastal area off Hokkaido in the northern Japan Sea. It was found that the 240Pu/239Pu atom ratios in the surface sediments for the Japanese side of the Japan Sea ranged from 0.15 to 0.22. The 240Pu/239Pu atom ratios in the surface sediments collected from the Yamato Basin and Japan Basin were typically close to the global fallout value of 0.18, but higher 240Pu/239Pu atom ratios (0.20-0.22) were seen in samples from the northern Japan Sea. Vertical profiles of 240Pu/239Pu atom ratios revealed that these atom ratios were higher than the global fallout value in the deeper layers in the sediment cores from the northern Japan Sea on the Hokkaido coast and the Yamato Basin. In addition, the sediment cores from the northern Japan Sea near the Soya Strait and in the Ishikari Estuary had a constant vertical distribution of 240Pu/239Pu atom ratios with a value around 0.20. The results obtained in this work support the hypothesis that the prevailing ocean currents transported Pu derived from the Pacific Proving Grounds to the Japan Sea, and this contributed to the general elevated Pu inventory in the Japan Sea. PMID- 15752503 TI - Heavy metals in Lake Balaton: water column, suspended matter, sediment and biota. AB - During the period 1999-2002, five sampling cruises have been carried out on Lake Balaton to assess trace metal distribution in the lake and to identify major sources. Eighteen elements, including Cr, Co, Ni, Cu, Zn, Cd, Pb (trace metals) and Al, Ba, Ca, Fe, K, Mg, Mn, Na, P, S, Sr (major metals), were determined in one or more of the lake's compartments. Lower trace metal concentrations in rainwater were observed in June and February 2000, while much higher levels were present in September 2001 (during a storm event) and in snow (February 2000). In the Northern and Western parts of the lake, especially at the inflow of river Zala and the locations of the yacht harbours, metal concentrations were higher in almost all compartments. Because the lake is very shallow, storm conditions also change significantly the metal distributions in the dissolved and particulate phases. The Kis-Balaton protection system located on Zala river functions very efficiently for retaining suspended particulate matter (SPM; 72% retention) and associated metals. Metal concentrations in surface sediments of the lake showed a high variability. After normalisation for the fine sediment fraction, only a few stations including Zala mouth appeared to be enriched in trace metals. In zooplankton, Zn seemed to be much more elevated compared to the other trace metals. Based on the molar ratios of the trace metals in the various compartments and input flows of the lake, several trends could be deduced. For example, molar ratios of the trace metals in the dissolved and solid (suspended particulate matter and sediments) phases in the lake are fairly similar to those in Zala River. PMID- 15752505 TI - Release of toxic metals and metalloids from Los Rueldos mercury mine (Asturias, Spain). AB - The abandoned mercury mining works of "Los Rueldos" are located 20 km from Oviedo, along the northwestern border of the Asturian Central Coal Basin, in an area with intense tectonization. Hg mainly appears as cinnabar, but occasionally metacinnabar and native Hg are present; associated with Hg ore appear As-rich minerals (arsenopyrite, As-rich pyrite, realgar and scorodite). In the spoil heap, Hg content ranges from 14 to 2224 mg kg(-1), and As from 4746 to 62,196 mg kg(-1). Mine drainage and spoil heap leachates show acidic conditions (pH: 2.43 2.50), 2900-4600 mg l(-1) sulphate, 1.4-9.2 mg l(-1) As, 0.03-0.48 mg l(-1) Pb, and 3.6-14 microg l(-1) Hg. According to the analytical data and characteristics of the site, the application of corrective measurements to avoid the dispersion of contaminants in the environment must be considered. PMID- 15752504 TI - Mercury distribution and speciation in Lake Balaton, Hungary. AB - The distribution and speciation of mercury in air, rain, lake water, sediment, and zooplankton in Lake Balaton (Hungary) were investigated between 1999 and 2002. In air, total gaseous mercury (TGM) ranged from 0.4 to 5.9 ng m(-3) and particulate phase mercury (PPM) from 0.01 to 0.39 ng m(-3). Higher concentrations of both TGM and PPM occurred during daytime. Higher concentrations of PPM occurred in winter. In rain and snow, total mercury ranged from 10.8 to 36.7 ng L(-1) in summer but levels up to 191 ng L(-1) in winter. Monomethylmercury (MMHg) concentrations ranged from 0.09 to 1.26 ng L(-1) and showed no seasonal variations. Total Hg in the unfiltered lake water varied spatially, with concentrations ranging from 1.4 to 6.5 ng L(-1). Approximately 70% of the total Hg is dissolved. MMHg levels ranged from 0.08 to 0.44 ng L(-1) as total and from 0.05 to 0.37 ng L(-1) in the dissolved form. Lower Hg concentrations in the water column occurred in winter. In suspended particulate matter and in sediment, total mercury ranged from 9 to 160 ng g(-1) dw, and MMHg ranged from 0.07 to 0.84 ng g( 1) dw. In zooplankton, an average mercury level of 31.0+/-6.8 ng g(-1) dw occurred, with MMHg accounting for approximately 17%. In sediments, suspended matter- and zooplankton-high Hg and MMHg levels occurred at the mouth of the River Zala, but, in the lake, higher concentrations occurred on the Northern side, and an increasing trend from north-west to north-east was observed. In general, regarding Hg, Lake Balaton can be considered as a relatively uncontaminated site. The high-pH and well-oxygenated water as well as the low organic matter content of the sediment does not favour the methylation of Hg. In addition, bioconcentration and bioaccumulation factors are relatively low compared to other aquatic systems. PMID- 15752506 TI - A comparison of total mercury and methylmercury export from various Minnesota watersheds. AB - Methylmercury (MeHg) bioaccumulates in aquatic food webs and can pose health risks to animals at higher trophic levels. Characterization of MeHg production in and export from watersheds can help clarify exposure scenarios for aquatic life downstream. A number of studies have demonstrated that anoxic conditions in the saturated soils of wetlands can promote the production of MeHg, and these wetlands may be major sources of MeHg to connected water bodies. Here, we report in-stream loadings of total mercury (THg) and MeHg for five rivers in Minnesota (USA). The watersheds of these rivers differ widely in the proportion of land area made up by wetlands and in other land use, drainage, and soil characteristics. Export of THg from these rivers varied widely, with much higher loadings and annual average concentrations of THg in streams of the Minnesota River basin compared to streams in the headwater Mississippi River basin. In contrast and despite the apparent differences in the makeup of these watersheds, yields and annual average concentrations of MeHg were remarkably similar for the rivers studied here. Differences in land use/land cover, drainage, soils, and other characteristics of these watersheds influence the export of both THg and MeHg in these rivers, but overall MeHg yields vary less than THg yields. PMID- 15752507 TI - Surface adsorption of metals onto the earthworm Lumbricus rubellus and the isopod Porcellio scaber is negligible compared to absorption in the body. AB - In terrestrial organisms, bioaccumulation is usually based on a summation of the amount of metal adsorbed to the body wall and absorbed into the body. The relative proportions of metal adsorption and absorption are usually not quantified. In this study, the distinction between adsorbed and absorbed metals was investigated in two different terrestrial species exposed to metals for 2 weeks. The earthworm Lumbricus rubellus was chosen as representative for organisms mainly taking up metals via the dermal route, and the isopod Porcellio scaber as an organism taking up metals mainly via the alimentary tract. Cross sections of whole animals were made using a cryostat and accumulated metals were localized by means of a phosphor screen (autoradiography). Radiolabels were used to determine the distribution of metals over the different organs and to distinguish between adsorption and absorption. Cd in the earthworm was mainly found in tissues of the chloragogenous region, whereas Zn was also found in various other organs and in the connective tissue. In the isopod, both Cd and Zn were mainly located in the hepatopancreas. Adsorbed amounts of Cd and Zn were negligible compared to internalized Cd and Zn concentrations for both organisms. Consequently, when focusing on effects of metal uptake for the organism itself, there is no need to correct for adsorption. This suggests that adsorption to the epidermis is not a rate limiting step in metal uptake by soil invertebrates. PMID- 15752509 TI - A continuum of care to save newborn lives. PMID- 15752510 TI - Epidemiological transition, medicalisation of childbirth, and neonatal mortality: three Brazilian birth-cohorts. PMID- 15752511 TI - Conceiving and dying in Afghanistan. PMID- 15752512 TI - Evidence-based: US road and public-health side of the street. PMID- 15752513 TI - Mucormycosis: an infectious complication of traumatic injury. PMID- 15752514 TI - Chimborazo and the old kilogram. PMID- 15752517 TI - Development. PMID- 15752518 TI - Controversial treatments for spinal-cord injuries. PMID- 15752519 TI - Controversial treatments for spinal-cord injuries. PMID- 15752520 TI - Evidence base for interventions in complex emergencies. PMID- 15752521 TI - The Kozijavkin method: giving parents false hope? PMID- 15752523 TI - Salt in food. PMID- 15752522 TI - Procurement of organs from executed prisoners. PMID- 15752524 TI - Doctors and bioterrorism. PMID- 15752525 TI - Salt in food. PMID- 15752526 TI - Poly-ticks, politics, and Lyme disease. PMID- 15752527 TI - A Doctor's Covenant to address a staff shortage. PMID- 15752528 TI - The challenge of reducing neonatal mortality in middle-income countries: findings from three Brazilian birth cohorts in 1982, 1993, and 2004. AB - BACKGROUND: Middle-income countries will need to drastically reduce neonatal deaths to achieve the Millennium Development Goal for child survival. The evolution of antenatal and perinatal care indicators in the Brazilian city of Pelotas from 1982 to 2004 provides a useful case study of potential challenges. METHODS: We prospectively studied three birth cohorts representing all urban births in 1982, 1993, and from January to July, 2004. The same methods were used in all three studies. FINDINGS: Despite improvements in maternal characteristics, prevalence of preterm births increased from 6.3% (294 of 4665) in 1982 to 16.2% (342 of 2112) in 2004, corresponding to a 47 g reduction in mean birthweight. Average number of antenatal visits in 2004 was 8.3 per woman, but quality of care was still inadequate--97% of women had an ultrasound scan, but only 1830 (77%) had a vaginal examination and 559 of 1748 non-immunised women did not receive tetanus toxoid. Rate of caesarean sections increased greatly, from 28% (1632 of 5914) in 1982 to 43% (1039 of 2403) in 2004, reaching 374 of 456 (82%) of all private deliveries in 2004. The increased rate of preterm births seemed to result largely from caesarean sections or inductions. Newborn care improved, and gestational-age-specific mortality rates had fallen by about 50% since 1982. As a result, neonatal mortality rates had been stable since 1990, despite the increase in preterm deliveries. INTERPRETATION: Excessive medicalisation--including labour induction, caesarean sections, and inaccurate ultrasound scans--led by an unregulated private sector with spill-over effects to the public sector, might offset the gains resulting from improved maternal health and newborn survival. These challenges will have to be faced by middle-income countries striving to achieve the child survival Millennium Development Goal. PMID- 15752529 TI - Incidence of macrolide resistance in Streptococcus pneumoniae after introduction of the pneumococcal conjugate vaccine: population-based assessment. AB - BACKGROUND: The prevalence of macrolide resistance in Streptococcus pneumoniae has risen in recent years after the introduction of new macrolides and their increased use. We assessed emergence of macrolide-resistant invasive S pneumoniae disease in Atlanta, GA, USA, before and after the licensing, in February 2000, of the heptavalent pneumococcal conjugate vaccine for young children. METHODS: Prospective population-based surveillance was used to obtain pneumococcal isolates and demographic data from patients with invasive pneumococcal disease. We calculated cumulative incidence rates for invasive pneumococcal disease for 1994-2002 using population estimates and census data from the US Census Bureau. FINDINGS: The incidence of invasive pneumococcal disease in Atlanta fell from 30.2 per 100,000 population (mean annual incidence 1994-99) to 13.1 per 100,000 in 2002 (p<0.0001). Striking reductions were seen in children younger than 2 years (82% decrease) and in those 2-4 years (71% decrease), age-groups targeted to receive pneumococcal conjugate vaccine. Significant declines were also noted in adults aged 20-39 (54%), 40-64 (25%), and 65 years and older (39%). Macrolide resistance in invasive S pneumoniae disease in Atlanta, after increasing steadily from 4.5 per 100,000 in 1994 to 9.3 per 100,000 in 1999, fell to 2.9 per 100,000 by 2002. Reductions in disease caused by mefE-mediated and erm-mediated macrolide resistant isolates of conjugate-vaccine serotypes 6B, 9V, 19F, and 23F, and the vaccine-associated serotype 6A were also recorded. INTERPRETATION: Vaccines can be a powerful strategy for reducing antibiotic resistance in a community. PMID- 15752530 TI - Where giving birth is a forecast of death: maternal mortality in four districts of Afghanistan, 1999-2002. AB - BACKGROUND: Maternal mortality in Afghanistan is uniformly identified as an issue of primary public-health importance. To guide the implementation of reproductive health services, we examined the numbers, causes, and preventable factors for maternal deaths among women in four districts. METHODS: We did a retrospective cohort study of women of reproductive age (15-49 years) who died between March 21, 1999, and March 21, 2002, in four selected districts in four provinces: Kabul city, Kabul province (urban); Alisheng district, Laghman province (semirural); Maywand, Kandahar province (rural); and Ragh, Badakshan province (rural, most remote). Deaths among women of reproductive age were identified through a survey of all households in randomly selected villages and investigated through verbal autopsy interviews of family members. FINDINGS: In a population of 90 816, 357 women of reproductive age died; 154 deaths were related to complications during pregnancy, childbirth, or the puerperal period. Most maternal deaths were caused by ante-partum haemorrhage, except in Ragh, where a greater proportion of women died of obstructed labour. All measures of maternal risk were high, especially in the more remote areas; the maternal mortality ratio (per 100,000 livebirths) was 418 (235-602) in Kabul, 774 (433-1115) in Alisheng, 2182 (1451-2913) in Maywand, and 6507 (5026-7988) in Ragh. In the two rural sites, no woman who died was assisted by a skilled birth attendant. INTERPRETATION: Maternal mortality in Afghanistan is high and becomes significantly greater with increasing remoteness. Deaths could be averted if complications were prevented through optimisation of general health status and if complications that occurred were treated to reduce their severity--efforts that require a multisectoral approach to increase availability and accessibility of health care. PMID- 15752531 TI - Relation between muscle Na+K+ ATPase activity and raised lactate concentrations in septic shock: a prospective study. AB - BACKGROUND: Hyperlactataemia during septic shock is often viewed as evidence of tissue hypoxia. However, this blood disorder is not usually correlated with indicators of perfusion or diminished with increased oxygen delivery. Muscles can generate lactate under aerobic conditions in a process linking glycolytic ATP supply to stimulation of Na+K+ ATPase. Using in-vivo microdialysis, we tested whether inhibition of Na+K+ ATPase can reduce muscle lactate. METHODS: In 14 patients with septic shock, two microdialysis probes were inserted into the quadriceps muscles and infused with lactate-free Ringer's solution in the absence or presence of 10(-7) mol/L ouabain, a specific inhibitor of Na+K+ ATPase. We measured lactate and pyruvate concentrations in both the dialysate fluid and arterial blood samples. FINDINGS: All patients had increased blood lactate concentrations (mean 4.0 mmol/L; SD 2.1). Lactate and pyruvate concentrations were consistently higher in muscle than in arteries during the study period, with a mean positive gradient of 1.98 mmol/L (SD 0.2; p=0.001) and 230 micromol/L (30; p=0.01), respectively. Ouabain infusion stopped over production of muscle lactate and pyruvate (p=0.0001). Muscle lactate to pyruvate ratios remained unchanged during ouabain infusion with no differences between blood and muscle. INTERPRETATION: Skeletal muscle could be a leading source of lactate formation as a result of exaggerated aerobic glycolysis through Na+K+ ATPase stimulation during septic shock. Lactate clearance as an end-point of resuscitation could therefore prove useful. RELEVANCE TO CLINICAL PRACTICE: In patients with septic shock, a high lactate concentration should be interpreted as a marker of disease, portending a bad outcome. The presence of hyperlactataemia in resuscitated septic patients should not be taken as proof of oxygen debt needing increases in systemic or regional oxygen transport to supranormal values. Lactate, instead of being regarded only as a marker of hypoxia, might be an important metabolic signal. PMID- 15752532 TI - Multifocal cutaneous mucormycosis complicating polymicrobial wound infections in a tsunami survivor from Sri Lanka. AB - A man injured in the tsunami of Dec 26, 2004, returned to Sydney for management of his soft-tissue injuries. Despite broad-spectrum antibiotics, surgical wound debridement, and vigilant wound care, his condition worsened. Muscle and fat necrosis developed in a previously debrided thigh wound, and necrotising lesions arose from previous abrasions. Histological analysis showed mucormycosis in three non-contiguous sites, and Apophysomyces elegans was isolated from excised wound tissue. Wound infections, both bacterial and fungal, will undoubtedly add to the morbidity and mortality already recorded in tsunami-affected areas. Other cases [correction] of cutaneous mucormycosis might develop in survivors, but this disease can be difficult to diagnose and even harder to treat, particularly in those remaining in affected regions. PMID- 15752533 TI - Sensorineural hearing loss in children. AB - During the past three to four decades, the incidence of acquired sensorineural hearing loss (SNHL) in children living in more developed countries has fallen, as a result of improved neonatal care and the widespread implementation of immunisation programmes. The overall decrease has been accompanied by a relative increase in the proportion of inherited forms of SNHL. The contribution made by one gene in particular, GJB2, to the genetic load of SNHL has strongly affected the assessment and care of children with hearing loss. These changes in the incidence of SNHL have not been seen in children living in less developed countries, where the prevalence of consanguinity is high in many areas, and both genetic and acquired forms of SNHL are more common, particularly among children who live in poverty. Focused genetic counselling and health education might lead to a decrease in the prevalence of inherited SNHL in these countries. Establishment of vaccination programmes for several vaccine-preventable infectious diseases would reduce rates of acquired SNHL. Although the primary purpose of such programmes is the prevention of serious and in many cases fatal infections, a secondary benefit would be a reduction in disease-related complications such as SNHL that cause permanent disability in survivors. PMID- 15752534 TI - 4 million neonatal deaths: when? Where? Why? AB - The proportion of child deaths that occurs in the neonatal period (38% in 2000) is increasing, and the Millennium Development Goal for child survival cannot be met without substantial reductions in neonatal mortality. Every year an estimated 4 million babies die in the first 4 weeks of life (the neonatal period). A similar number are stillborn, and 0.5 million mothers die from pregnancy-related causes. Three-quarters of neonatal deaths happen in the first week--the highest risk of death is on the first day of life. Almost all (99%) neonatal deaths arise in low-income and middle-income countries, yet most epidemiological and other research focuses on the 1% of deaths in rich countries. The highest numbers of neonatal deaths are in south-central Asian countries and the highest rates are generally in sub-Saharan Africa. The countries in these regions (with some exceptions) have made little progress in reducing such deaths in the past 10-15 years. Globally, the main direct causes of neonatal death are estimated to be preterm birth (28%), severe infections (26%), and asphyxia (23%). Neonatal tetanus accounts for a smaller proportion of deaths (7%), but is easily preventable. Low birthweight is an important indirect cause of death. Maternal complications in labour carry a high risk of neonatal death, and poverty is strongly associated with an increased risk. Preventing deaths in newborn babies has not been a focus of child survival or safe motherhood programmes. While we neglect these challenges, 450 newborn children die every hour, mainly from preventable causes, which is unconscionable in the 21st century. PMID- 15752535 TI - The 21st century health challenge of slums and cities. PMID- 15752536 TI - 5 NICE years. PMID- 15752537 TI - Complication of urethral catheterisation. PMID- 15752538 TI - Traumatic scoliosis. PMID- 15752539 TI - Chronic ethanol intake modifies renin-angiotensin system-regulating aminopeptidase activities in mouse cerebellum. AB - In developing cerebellum, where critical periods of vulnerability have been established for several basic substances, it has been extensively studied the wide array of abnormalities induced by exposure to ethanol (EtOH). However, little is known about the effects of EtOH consumption on cerebellar functions in adult individuals. Several studies show participation in cognitive activities to be concentrated in the lateral cerebellum (hemispheres), whereas basic motor functions such as balance and coordination are represented in the medial parts of the cerebellum (vermis and paravermis). In addition to the circulating renin angiotensin system (RAS), a local system has been postulated in brain. The effector peptides of the RAS are formed via the activity of several aminopeptidases (AP). The present work analyses the effect of chronic EtOH intake on the RAS-regulating AP activities in the soluble and membrane-bound fractions of two cerebellar locations: the hemispheres and the vermis. We hypothesize that cerebellar RAS is involved in basic motor functions rather than in cognitive activities. PMID- 15752540 TI - Adrenomedullin gene expression and levels in the cardiovascular system after treatment with lipopolysaccharide. AB - To study the effect of septicaemia, the temporal changes in tissue adrenomedullin (AM) and preproAM mRNA levels were studied in the heart and blood vessels after lipopolysaccharide (LPS) injection. Radioimmunoassay and solution hybridization RNase protection assays were used to follow the changes in AM and its mRNA levels respectively after intraperitoneal injection of 10 mg/kg LPS in rats. The preproAM mRNA levels increased at 1 h in the right atrium after LPS injection, while the AM contents decreased at 1 h in the left atrium. The preproAM mRNA levels increased at 3 and 6 h in the left ventricle, whereas it increased at 6 h in the right ventricles after LPS injection. There was an increase in preproAM mRNA levels at 1 and 3 h in the mesenteric artery, while AM levels were increased at 1, 3 and 6 h. However, there were no such changes in the thoracic aorta. There were also increases in tumour necrosis factor-alpha (TNF-alpha), interleukin (IL) 1beta and IL-6 in the heart, and in the mesenteric artery (TNF-alpha and IL 1beta) and in thoracic aorta (IL-1beta and IL-6). The present results suggest that the biosynthesis and secretion of AM may be increased in cardiovascular tissues of rats injected with LPS, and that AM may play multiple roles in inflammation. PMID- 15752541 TI - Central penetration and stability of N-terminal tripeptide of insulin-like growth factor-I, glycine-proline-glutamate in adult rat. AB - Insulin-like growth factor-I is a neurotrophic factor and can prevent neurons from ischemic brain injury. However, the large molecular weight and metabolic effects can be problematic in its central delivery. Glycine-proline-glutamate (GPE) is the N-terminal tripeptide of insulin-like growth factor-I, which is naturally cleaved in the plasma and brain tissues. GPE reduces neuronal loss from hypoxic-ischemic brain injury following central administration. Central penetration and the stability of GPE in the plasma and central nervous system were examined in rats using radioimmunoassay and HPLC. GPE was rapidly metabolised in the plasma (8 min) after intraperitoneal administration. Despite having a short half-life in plasma, GPE was detected in the cerebrospinal fluid up to 40 min after intraperitoneal administration. With present of peptidase inhibitors, GPE existed in the brain tissue up to 3 h after intracerebroventricular administration, suggesting a role for peptolysis in its stability. The endopeptidase inhibitors 4- (2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) reduced GPE metabolism in the brain tissue while acid peptidase inhibitor pepstatin-A decreased GPE metabolism in the plasma. GPE reduced neuronal loss in the CA1-2 sub-region of the hippocampus given (intraperitoneally) after 30 min of hypoxic-ischemic injury in adult rats, further suggested the effectiveness of GPE central uptake. These results indicated that GPE crosses the blood-CSF and the functional CSF-brain barriers. The longer half-life of GPE in the CNS may be due to its unique enzymatic stability. PMID- 15752542 TI - Myocardial infarction stimulates galanin expression in cardiac sympathetic neurons. AB - Cardiac ischemia-reperfusion alters sympathetic neurotransmission in the heart, but little is known about its effect on neuropeptide expression in sympathetic neurons. Ischemia followed by reperfusion induces the production of inflammatory cytokines in the heart, including interleukin-6 and cardiotrophin-1. These cytokines and related molecules inhibit the expression of neuropeptide Y (NPY), and stimulate the expression of vasoactive intestinal peptide (VIP), substance P (SubP), and galanin (GAL) in cultured sympathetic neurons. Therefore, we quantified NPY, VIP, SubP, and GAL mRNA in neurons of the stellate ganglia 1 week after ischemia-reperfusion to determine if neuropeptide expression was altered in cardiac sympathetic neurons. NPY, VIP, and SubP mRNAs were unchanged compared to unoperated control animals, but GAL mRNA was increased significantly. The increased GAL mRNA was not accompanied by elevated GAL peptide content in the stellate ganglia. Galanin content was increased significantly in the heart, however, indicating that elevated GAL mRNA led to increased peptide production. GAL content was increased in the left ventricle below the coronary artery ligation, but was not increased significantly in the atria or the base of the heart above the ligation. The buildup of GAL specifically in the damaged left ventricle is consistent with previous reports that GAL is transported to regenerating nerve endings after axon damage. PMID- 15752543 TI - Effect of humanin on decreased ATP levels of human lymphocytes harboring A3243G mutant mitochondrial DNA. AB - Humanin (HN) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis by mutant Alzheimer's disease genes. This 24 residue peptide has been recently shown to suppress apoptosis by interfering with activation of Bcl-2-associated X protein (Bax) in cytosol. In the present study, we showed that HN increases ATP levels in human lymphocytes, muscular TE671 cells, and neural SKN-MC cells, and protects these cells from serum deprivation induced apoptosis. The suppressed apoptotic death of serum-deprived cells would be explained by the anti-Bax effect of HN; however, HN also increased ATP levels of serum-supplemented cells (non-apoptotic cells), in which Bax is likely to be inactive. This result suggests the presence of a certain mechanism independent of Bax inactivation to increase ATP levels of cells under non-apoptotic condition. By treatment with HN, the ATP levels of lymphocytes from patients with mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) associated with A3243G mutant mtDNA were increased as well, suggesting that HN is able to prevent cells in MELAS from falling into ATP deficiency. Our quantitative PCR findings indicated that the HN-induced increase in ATP may not be a consequence of mitochondrial proliferation, because HN rather suppressed mtDNA replication. This suppression may be important in the treatment of affected cells in MELAS, since the mutant mtDNAs that increase during compensatory mtDNA replication for ATP deficiency cause excessive formation of reactive oxygen species, leading to further energy crisis. We thus propose that HN, which increases cellular ATP levels without inducing mtDNA replication, may be suited for the treatment of MELAS. PMID- 15752545 TI - Intracerebroventricular administration of galanin decreases free water intake and operant water reinforcer efficacy in water-restricted rats. AB - The 29/30 amino acid neuropeptide galanin coexists with vasopressin in the hypothalamus and has been shown to inhibit the actions of vasopressin and aldosterone, suggesting an inhibitory role for galanin in physiological water retention mechanisms and water seeking and water consumption behavior. Little work, however, has examined a role for galanin in water intake regulation. Furthermore, many experiments that have reported galanin-induced impairments in the performance of tasks thought to measure learning and memory have used water restriction routines and water reinforcers to maintain responding. Therefore, the present study examined the effects of intracerebroventricularly administered galanin (5.0-20.0 microg/5 microl) on free water consumption during a 10 min test session and a follow up open field exploration, an operant progressive ratio (PR) schedule, a test used to assess reinforcer strength, and an operant fixed time schedule (FT 20) in 23.5h water restricted rats. Finally, in an additional experiment that was designed to simulate the effects of a galanin-induced decrease in water reinforcer efficacy, the rats were allowed access to water prior to testing in an operant delayed non-matching to position (DNMTP) task. A galanin-induced decrease in water consumption was observed in both the free access test and the FT 20 at the 20 microg dose, but no significant galanin induced alterations in open field behavior. A decrease in responses emitted and rewards received was observed on the PR schedule at the 5, 10, and 20 microg doses. Pre-session access to water significantly reduced the number of trials per session in the DNMTP but did not reduce accuracy. This study is the first to observe a galanin-induced reduction in water intake and reinforced operant behavior, and suggests that galanin may play a role in regulating water intake and reinforcement. However, the present data also suggest that DNMTP choice accuracy deficits observed previously cannot be attributed to a galanin-induced change in reinforcer efficacy. PMID- 15752544 TI - Acute, subacute and chronic effects of central neuropeptide Y on energy balance in rats. AB - Central neuropeptide Y (NPY) injection has been reported to cause hyperphagia and in some cases also hypometabolism or hypothermia. Chronic central administration induced a moderate rise of short duration in body weight, without consistent metabolic/thermal changes. In the present studies the acute and subsequent subacute ingestive and metabolic/thermal changes were studied following intracerebroventricular (i.c.v.) injections of NPY in cold-adapted and non adapted rats, or the corresponding chronic changes following i.c.v. NPY infusion. Besides confirming basic earlier data, we demonstrated novel findings: a temporal relationship for the orexigenic and metabolic/thermal effects, and differences of coordination in acute/subacute/chronic phases or states. The acute phase (30-60 min after injection) was anabolic: coordinated hyperphagia and hypometabolism/hypothermia. NPY evoked a hypothermia by suppressing any (hyper)metabolism in excess of basal metabolic rate, without enhancing heat loss. Thus, acute hypothermia was observed in sub-thermoneutral but not thermoneutral environments. The subsequent subacute catabolic phase exhibited opposite effects: slight increase in metabolic rate, rise in body temperature, reaching a plateau within 3-4 h after injection -- this was maintained for at least 24 h; meanwhile the food intake decreased and the normal daily weight gain stopped. This rebound is only indirectly related to NPY. Chronic (7-day long) i.c.v. NPY infusion induced an anabolic phase for 2-3 days, followed by a catabolic phase and fever, despite continued infusion. In cold-adaptation environment the primary metabolic effect of the infusion induced a moderate hypothermia with lower daytime nadirs and nocturnal peaks of the circadian temperature rhythm, while at near thermoneutral environments in non-adapted rats the infusion attenuated only the nocturnal temperature rise by suppressing night-time hypermetabolism. Further finding is that in cold-adapted animals, the early feeding effect of NPY-infusion was enhanced, whereas the early hypothermic effect in cold was limited by interference with competing thermoregulatory mechanisms. PMID- 15752546 TI - Armadillidin: a novel glycine-rich antibacterial peptide directed against gram positive bacteria in the woodlouse Armadillidium vulgare (Terrestrial Isopod, Crustacean). AB - We report the isolation and the characterization of a novel antibacterial peptide from hemocytes of the woodlouse Armadillidium vulgare, naturally infected or uninfected by Wolbachia, an intracellular Gram-negative bacterium. This molecule displays antibacterial activity against Gram-positive bacteria despite its composition which classes it into the glycine-rich antibacterial peptide family, usually directed against fungi and Gram-negative bacteria. The complete sequence was determined by a combination of Edman degradation, mass spectrometry and cDNA cloning using a hemocyte library. The mature peptide (53 residues) has a 5259 Da molecular mass and is post-translationally modified by a C-terminal amidation. This peptide is characterized by a high level of glycine (47%) and a fivefold repeated motif GGGFH(R/S). As no evident sequence homology to other hitherto described antibacterial peptides has been found out, this antibacterial peptide was named armadillidin. Armadillidin is constitutively expressed in hemocytes and appears to be specific of A. vulgare. PMID- 15752547 TI - Pathogenesis and inflammatory response to Edwardsiella tarda infection in the zebrafish. AB - The zebrafish (Danio rerio) is a widely used model for developmental biology, neurobiology, toxicology, and genetic disease. Recently, the zebrafish has been recognized as a valuable model for infectious disease and immunity. In this study the pathogenesis and inflammatory cytokine response of zebrafish to experimental Edwardsiella tarda infection was characterized. In challenge experiments, zebrafish embryos were susceptible to infection by immersion. Adult fish were susceptible to challenge by intraperitoneal (ip) injection but not static immersion unless the epithelial layer was perturbed by scraping prior to exposure. To determine if E. tarda infection induces a typical acute inflammatory response, mRNA expression levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha) were assessed by quantitative real-time PCR. The expression levels of IL-1beta and TNFalpha were significantly upregulated in infected zebrafish embryos and adults. The methods developed in this study will be particularly valuable for targeted gene disruption studies of host immune components and in zebrafish genetic screens. PMID- 15752548 TI - Expression profiling of immune-related genes from Japanese flounder Paralichthys olivaceus kidney cells using cDNA microarrays. AB - A Japanese flounder Paralichthys olivaceus cDNA microarray containing 871 unique cDNAs including 91 putative immune-related genes from our EST studies was constructed and used to characterize of gene expression of in vitro grown kidney cells stimulated with mitogens such as ConA, PMA, LPS or infected with hirame rhabdovirus (HRV). The numbers of genes whose expressions were increased or decreased by these factors were: 17 by Con A, 139 by PMA, 76 by LPS and 182 by HRV infection. The treatment of Con A for 1 and 6h affected the expression of only a few of the immune-related genes. PMA down-regulated far more genes than it up-regulated. Apoptosis-related factors, such as c-fos, NGF induced protein IB and NR13 genes, were among the genes whose expressions were induced by PMA. LPS induced the expression of inflammation-related genes, such as IL-1beta, monocyte chemotactic protein 1 and collagenase. The expressions of many genes were induced after 3h HRV infection but some of them were decreased to the basal level after 6h HRV infection. The expression of some genes of unknown function were induced or reduced by Con A, PMA or LPS or by HRV infection in different time periods. From all of the gene expression profiling in this study, we could get lots of information about the dynamic changes in the gene expression of the kidney cells under different stress or stimulations. PMID- 15752549 TI - Adhesion and ingestion activities of fish phagocytes induced by bacterium Aeromonas salmonicida can be distinguished and directly measured from highly diluted whole blood of fish. AB - The phagocytes of fish play an important role in innate host defense against bacterial infection, and participate in various immunoregulatory processes. Here, we investigated the effects of various opsonins in the ingestion and adhesion processes by examining respiratory burst (RB) activity in blood and head kidney (HK) fish phagocytes. RB activity was induced in rainbow trout phagocytes with the bacterium Aeromonas salmonicida (strain MT004) in the presence of various opsonins [purified antibodies (Ab), immune serum (IS), normal serum (NS) and heat inactivated immune serum (HI-IS)], and measured in terms of luminol-amplified chemiluminescence (CL) emission at 20 degrees C for 210 min. The RB activity of blood phagocytes was measured directly from highly diluted whole blood and compared to that observed in isolated head kidney (HK) phagocytes measured under similar conditions. In addition, the extracellular RB activity of adhesion (extracellular degranulation) and the intracellular RB activity of ingestion were distinguished through their inhibition by gelatin and cytochalasin D. Our results showed that the first CL peak appeared within 50 min, and decreased or vanished when gelatin was added to the reaction or when the active complement was destroyed by heating. The second CL peak appeared after 50 min, depending on the utilized opsonin, and vanished when cytochalasin D was added to the reaction. Our results indicate that adhesion and ingestion compete for consumption of reactive oxygen intermediates. Specific IgM without an active complement was a relatively inefficient opsonin, whereas specific IgM with an active complement increased the magnitude of ingestion-mediated RB activity and accelerated the ingestion of target bacteria. Taken together, these results indicate that adhesion and ingestion responses competed for limited phagocyte resources and that the bacterial uptake by blood phagocytes can be measured directly from highly diluted blood. PMID- 15752550 TI - Lymphoid follicles of different phenotype appear in ileum during involution of the sheep ileal Peyer's patch. AB - The ileal Peyer's patch (IPP) of young sheep is a site of diversification of the primary antibody repertoire and where involution takes place at young age. Tissue samples from the ileum were collected in 134 animals aged from 1 month to 6 years, and IPP follicle phenotypes were characterised. We describe a new type of ileal lymphoid follicles that became relatively more frequent during involution, and had numerous intrafollicular T-cells and BAQ44A+ B-cells and large interfollicular T-cell areas. As opposed to classical IPP follicles in which the BAQ44A+ cells were confined to the narrow follicle-neck region, the novel atypical ileal lymphoid follicle had these cells distributed throughout the follicle. The relative distribution of cell types in the typical IPP follicle remained fairly constant during involution. Many animals older than 9 months (64/92) still had had typical IPP follicles and even sheep 4 years and older (5/9) had IPP-type follicles. PMID- 15752551 TI - Kinetics and distribution of bovine gammadelta T-lymphocyte in the intestine: gammadelta T cells accumulate in the dome region of Peyer's patch during prenatal development. AB - The kinetics and distribution of gammadelta T cells in bovine intestine including jejunal and ileal Peyer's patch were examined. The number of gammadelta T cells increased significantly in the dome region during prenatal development, but decreased notably after birth. The number of some gammadelta T cells, CD4+ cells, and CD8+ cells in the intestinal villi remained constant during prenatal development, but increased significantly after birth. The kinetics of the gammadelta T cells in the dome region during prenatal development were quite distinct from those of the gammadelta T cells, CD4+ cells, and CD8+ cells in the intestinal villi. In the fetal ileum at full-term gestation, the frequencies of expression of the T-cell receptor gamma variable region (TCR Vgamma) family were TCR Vgamma1 (48%), Vgamma2 (4%), and Vgamma5 (48%). However, in 2-month-old calf ileum, TCR Vgamma5 (90%) was dominant. We speculate that functional differences exist between gammadelta T cells in the dome region during prenatal development and in the intestinal villi after birth. PMID- 15752552 TI - Use of a microgravity organ culture dish system to demonstrate the signal dampening effects of modeled microgravity during T cell development. AB - Recently, we have shown that exposure of fetal thymus organ cultures (FTOC) to modeled microgravity (MMG) using a clinostat with a microgravity organ culture dish system (MOCDS) blocks T cell development in a manner independent of steroid stress hormones present in vivo. In this study, we describe the development of the MOCDS system, as well as its use in attempting to understand the mechanism by which T cell development is inhibited in MMG. We show that after MMG exposure FTOC exhibited a significant reduction in CD4+CD8+ double positive (DP) cell production, but those DP cells which remained expressed higher levels of the T cell receptor (TCR) associated molecule, CD3. Interestingly, CD4-CD8- double negative (DN) cells expressed lower levels of CD3 on their surface. DN, as well as immature single positive (ISP) cells, also expressed reduced levels of the IL 7 receptor alpha chain (CD127). These changes in CD3 and CD127 expression were concomitantly associated with an increased production of tumor necrosis factor (TNF)-alpha. We were also able to show that addition of an exogenous signal (anti CD3epsilon monoclonal antibody) to these cultures effectively mitigated the MMG induced effects, suggesting that MMG-exposure causes a signal dampening effect on developing thymocytes. PMID- 15752553 TI - T cells at the foot of the mountains: 10 years of EFIS-Tatra Conferences. PMID- 15752554 TI - The emerging role of the T cell-specific adaptor (TSAd) protein as an autoimmune disease-regulator in mouse and man. AB - T cell-specific adapter protein is a relatively recently described signaling adapter molecule expressed predominantly in T cells and NK cells. Studies in mouse and man have indicated that reduced expression of TSAd in T cells may predispose toward the development of autoimmune disease. In lupus-prone TSAd deficient mice the development of autoimmunity is associated with an impaired T cell death response to antigens in vivo. Probably, this impaired death response is consequent to reduced T cell antigen receptor (TCR)-induced synthesis of the interleukin-2 (IL-2) cytokine in TSAd-deficient T cells. TSAd appears to contribute to IL-2 synthesis at multiple different levels acting in both the nucleus and cytoplasm of T cells. Recent advances relating to the role of TSAd in T cell signal transduction and as a regulator of autoimmune responses are discussed. PMID- 15752555 TI - Complement: central to innate immunity and bridging to adaptive responses. AB - The complement system, a pillar of innate immunity, has belatedly become recognised as a key modulator of adaptive immunity, acting to direct, modulate and modify the responses of lymphocytes to stimuli. These effects are mediated by interactions between complement components or activation-derived fragments and specific binding proteins--complement receptors and regulators--on the target cells. This review will describe the current state of knowledge in this swiftly moving field. It is hoped that the recognition of these properties will help to establish complement in the role it richly deserves as the lynchpin of immunity. PMID- 15752556 TI - Mucosal adjuvants and anti-infection and anti-immunopathology vaccines based on cholera toxin, cholera toxin B subunit and CpG DNA. AB - Mucosal immunisation may be used both to protect the mucosal surfaces against infections and as a means for immunological treatment of peripheral immunopathological disorders through the induction of systemic antigen-specific tolerance ('oral tolerance'). The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems that can help to present the appropriate vaccine or immunotherapy antigens to the mucosal immune system. The most potent (but also toxic) mucosal adjuvants are cholera toxin (CT) and the closely related Escherichia coli heat-labile enterotoxin (LT), and much effort and significant progress have been made recently to generate toxicologically acceptable derivatives of these toxins with retained adjuvant activity. Among these are the non-toxic, recombinantly produced cholera toxin B-subunit (CTB). CTB is a specific protective antigen component of a widely registered oral cholera vaccine as well as a promising vector for either giving rise to mucosal anti-infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. CT and CTB have also recently been used as combined vectors and adjuvants for markedly promoting ex vivo dendritic cell (DC) vaccination with different antigens and also steering the immune response to the in vivo-reinfused DCs towards either broad Th1 + Th2 + CTL immunity (CT) or Th2 or tolerance (CTB). Another type of mucosal adjuvants is represented by bacterial DNA or synthetic oligodeoxynucleotides containing CpG-motifs, which especially when linked to CTB have been found to effectively stimulate both innate and adaptive mucosal immune responses. The properties and clinical potential of these different classes of adjuvants are being discussed. PMID- 15752557 TI - Cure of innate intestinal immune pathology by CD4+CD25+ regulatory T cells. AB - CD4+CD25+ regulatory T (T(R)) cells are a naturally occurring population of T cells that suppress the development of a variety of pathological immune responses. However, as human inflammatory diseases are usually not diagnosed until after the onset of clinical symptoms, it is of great interest to determine whether CD4+CD25+ T(R) cells can reverse established pathology. To examine this question we have utilized a murine model of human inflammatory bowel disease (IBD), where pathology is triggered by infection of immune deficient RAG-/- mice with the pathogenic bacterium Helicobacter hepaticus. Here we demonstrate that adoptively transferred CD4+CD25+ T(R) cells can cure established intestinal inflammation that is mediated by innate immune activation in H. hepaticus infected RAG-/- mice. CD4+CD25+ T(R) cell-mediated amelioration of innate intestinal pathology was accompanied by a reversal in systemic innate immune activation, but did not involve any detectable anti-bacterial effects, as bacterial colonization levels were unchanged. Cure of established pathology was not achieved using subpopulations of CD4+CD25- T cells, further emphasizing the enhanced regulatory activity of CD4+CD25+ T(R) cells. PMID- 15752558 TI - From toxin to adjuvant: basic mechanisms for the control of mucosal IgA immunity and tolerance. AB - We provide compelling evidence that delivery of Ag in the absence of ADP ribosylation can promote tolerance, whereas ADP-ribosyltransferase activity induces IgA immunity and prevents tolerance. By linking Ag to the ADP ribosylating enzyme, cholera toxin subunit A1 (CTA1), we could show that the combination of targeting to antigen-presenting cells (APC) and enzymatic activity is a highly effective means of controlling the induction of tolerance or immunity. Firstly, we demonstrated that cholera toxin (CT), although potentially binding to all nucleated cells, in fact, bound preferentially to dendritic cells (DC) in vivo. Following injection of CT-conjugated Ag, we found that DC in the marginal zone (MZ) of the spleen accumulated Ag, a process that was GM1 ganglioside receptor dependent. Contrary to CTB, which also delivered Ag to the MZ DC, CT matured and activated co-stimulatory functions in the targeted DC and greatly augmented immune responses to Ag. Secondly, when Ag was incorporated into the CTA1-DD fusion protein, which equals the CT in adjuvant function but lacks GM1-ganglioside-binding ability, we greatly augmented specific responses to Ag. The DD-bound Ag was distinctly targeted to B cells and probably also to follicular dendritic cells (FDC) in vivo. Thus, in both constructs Ag was targeted to APC and associated with an ADP-ribosylating enzyme, which resulted in greatly enhanced immunogenicity. When the enzymatic activity was absent, as in CT B-subunit (CTB) or in the inactive CTA1R7K-DD mutant, Ag largely failed to stimulate an active immune response. Rather, this type of Ag exposure resulted in Ag-specific tolerance, especially when mucosal delivery of Ag was attempted. Therefore, targeting to APC in the absence or presence of the CTA1-enzyme appears to be an effective means to control tolerance and active protective IgA immunity. PMID- 15752559 TI - Molecular mechanisms that set the stage for DC-T cell engagement. AB - The unsurpassed capacity of dendritic cells (DC) to prime naive T cells is thought to depend on the formation of an immunological synapse. DC-SIGN, a C-type lectin exclusively expressed at the cell surface of DC, functions as an adhesion receptor facilitating T cell binding and priming through recognition of glycosylated ICAM-3 on naive T cells. Yet, DC-SIGN also mediates binding to pathogens such as HIV by recognizing glycosylated gp120. The scope of the present study was to investigate whether DC-SIGN upon recognition of its cellular ligand and pathogenic ligand affects DC synapse formation and activation/mobilization of other adhesion receptors such as LFA-1 to the cell contact site. Using a DC-SIGN deletion mutant, we show that DC-SIGN is a constitutively active receptor that mediates ligand binding independent of signaling through the cytoplasmic domain. Surprisingly, initial binding of gp120 to DC-SIGN did not result in increased adhesion levels of LFA-1 to its ligand ICAM-1 in both immature DC and Raji-DC SIGN cells. However, ligand binding to DC-SIGN induced recruitment of LFA-1 to the adhesion site. Moreover, we could demonstrate that activation of LFA-1 results in DC-SIGN-LFA-1 co-clustering in the cell membrane. This triggers binding of ligands to LFA-1 that are shared with DC-SIGN, such as ICAM-3, but not of ligands that are not shared with DC-SIGN, such as ICAM-1. Thus, we propose that upon ligand binding DC-SIGN recruits LFA-1 to the contact site, resulting in the formation of DC-SIGN-LFA-1 co-clusters, in which the initial DC-SIGN-mediated interactions with ligand are transient and eventually shift to more stable LFA-1 dependent interactions. PMID- 15752560 TI - HIV and human complement: inefficient virolysis and effective adherence. AB - Both, HIV envelope proteins gp120 and gp41 can directly activate complement system, even in the absence of HIV-specific antibodies. During the budding process HIV acquires host membrane-associated molecules among these complement regulatory proteins (CRPs). The presence of CRPs on the viral surface rescues HIV from complement-mediated virolysis. The inefficient virolysis results in the deposition of complement-fragments on the viral surface allowing interactions of HIV with complement receptor expressing cells. In this review, the interaction of HIV with the complement system and the consequences of complement opsonisation on virus infection will be discussed. PMID- 15752561 TI - Progression to CMV end-organ disease in HIV-1-infected individuals despite abundance of highly differentiated CMV-specific CD8+ T-cells. AB - Since CMV-specific T-cells have been shown to generally express an advanced state of differentiation, we investigated whether these mature CMV-specific T-cells are sustained in HIV-infected patients, who are not treated with HAART, receive no CMV medication, but do progress to AIDS with CMV end-organ disease (AIDS-CMV). CD8+ and CD4+ T-cell phenotype was studied in these patients in comparison with long-term asymptomatic HIV-infected individuals, progressors to AIDS without CMV end-organ disease as well as CMV-seropositive HIV-negative controls. CMV-specific CD8+ T-cells from progressors to AIDS-CMV expressed markers typical of highly differentiated effector T-cells, being CCR7-, CD27- CD45RO+/-, with high CD57 expression and increased Ki67 expression, compatible with functional effector cell capabilities. In addition, CD4+ T-cells with the characteristic CD27-CD28- phenotype previously shown to be induced by CMV infection specifically, were found in very high numbers in the HIV+ individuals, but the highest in progressors to AIDS-CMV just before onset of disease. Also the normally rare CD45RO-CD27-CD4+ subset increased significantly, whereas the CD45RO-CD27+CD4+ subset decreased. Our data show that in patients progressing to AIDS-CMV, CMV specific CD8+ T-cells have expanded and are fully differentiated to mature functional effector T-cells. These cells are not protective apparently, but may contribute to tissue-associated immunopathology characteristic of these clinical conditions. PMID- 15752562 TI - Newly identified host factors modulate HIV replication. PMID- 15752563 TI - The p95-100 kDa ligand of the T cell-specific adaptor (TSAd) protein Src-homology 2 (SH2) domain implicated in TSAd nuclear import is p97 Valosin-containing protein (VCP). AB - T cell-specific adapter protein (TSAd) is required for normal T cell antigen receptor (TCR)-induced transcription of cytokine genes in T cells. How TSAd controls cytokine transcription is unknown. Previously, we have shown that TSAd is actively transported to the nucleus of T cells suggesting that this adapter may in part function within this cellular compartment. Nuclear translocation of TSAd is dependent upon an intact Src-homology-2 (SH2) domain and a p95-100 kDa ligand of the SH2 domain has been implicated in nuclear import. Here, using microchemical techniques, we identify p95-100 as p97 Valosin-containing protein (VCP) whose homolog in yeast is the cell division control protein, CDC48. Physical interaction between TSAd and VCP can be demonstrated between endogenous proteins in T cells. Interaction is direct and is dependent upon phosphorylation of tyrosine residue 805 of VCP that has been previously recognized as a major target of tyrosine kinase(s) involved in TCR signaling. Significantly, with the use of CDC48 mutant yeast, we demonstrate that VCP/CDC48 is required for transport of TSAd into the eukaryotic nucleus. These findings provide important insights into the mechanism of TSAd nuclear import and the role of TSAd in T cell signal transduction. PMID- 15752564 TI - Can tissue transglutaminase be a marker of idiopathic inflammatory myopathies? AB - In the normal striated muscle, tissue transglutaminase (TG2) content is vestigial. However, this protein's presence has been reported to occur in myoblasts and myotubes during the fetal period. Its increased expression has been also found in the muscle tissue in the course of sporadic inclusion body myositis, as well as in polymyositis (PM) and dermatomyositis (DM), which are considered to be diseases of immunological origin. Based on in vitro studies, a substantial TG2 role in the infiltration of some T cell subsets into inflamed tissues has been suggested lately. In this study, the immunohistochemical reactions in the guinea pig experimental myositis specimens and in the ones from PM/DM patients were compared. The guinea pig tissue specimens were taken from muscles affected by experimental myositis induced by intramuscular injections of: 1/sera from 30 neoplasm patients with no metastases; 2/sera from 10 healthy people; 3/sera from 2 DM patients; 4/neuropeptides (SP, NPY or VIP) and from 5/the muscles affected by the reversed passive Arthus reaction (RPAR). The immunostaining for TG2 revealed substantial presence of this protein in single, damaged muscle fibers and a weak reaction in regenerating fibers appearing in PM/DM patients' specimens. From among experimental myositis specimens, a very intensive reaction appeared only in the damaged and regenerating muscle fibers present in the slides from guinea pig muscles injected with DM patients' sera. Such results suggest some presence of a specific factor(s) (the one(s) responsible for TG2 expression in the damaged muscle fibers) in DM patients' sera. The results suggest that transglutaminase can be a marker of inflammatory myopathies. A probable correlation between TG2 expression in muscles and organismal immunological factors, including the complement activation status, requires additional studies. PMID- 15752565 TI - Immune response after adjuvant mucosal immunization of mice with inactivated influenza virus. AB - Satisfactory mucosal immunity in the respiratory tract is very important for protection against influenza. It can be achieved only by mucosal immunization. Mucosal vaccination with inactivated influenza virus may not be sufficiently effective and suitable adjuvants are therefore sought. We tested intratracheal immunization of mice with inactivate B type influenza virus in a mixture with formolized G+ bacterium Bacillus firmus, whose adjuvant effects have previously been documented in another system. The treatment resulted in a marked increase of both systemic and mucosal antibody response in IgG and IgA classes. Stimulation of T lymphocytes after adjuvant immunization was very mild, no proliferation taking place after specific stimulation with antigen in vitro. However, slightly increased systemic (spleen) and local (lungs) production of cytokines without perceptible Th1/Th2 polarization was determined. B. firmus is an efficient adjuvant in respiratory tract immunization while with subcutaneous immunization it lowers the antibody response. PMID- 15752566 TI - Stereoselective differentiation in the Salt-induced Peptide Formation reaction and its relevance for the origin of life. AB - All living organisms on earth are almost totally made up of biomolecules of only one chiral form. For example, proteins are built almost exclusively of L-amino acids, and sugars are composed of D-saccharides, a fact that is usually referred to as biohomochirality. Its origin is the center of numerous investigations and theories but is not really elucidated yet. The results of experimental investigations of peptide formation in a prebiotically relevant scenario, as described in this paper, give indications on a possible pathway for the synthesis of homochiral L-peptides in the course of the Salt-induced Peptide Formation (SIPF) reaction. PMID- 15752567 TI - pH- and thermal-dependent conformational transition of PGAIPG, a repeated hexapeptide sequence from tropoelastin. AB - The secondary structure of PGAIPG (Pro-Gly-Ala-IIe-Pro-Gly), a repeated hexapeptide of tropoelastin, in buffer solution of different pH was determined by using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. The thermal-dependent structural change of PGAIPG in aqueous solution or in solid state was also examined by thermal FTIR microspectroscopy. The conformation of PGAIPG in aqueous solution exhibited a pH-dependent structural characterization. A predominant peak at 1614 cm(-1) (aggregated beta sheet) with a shoulder near 1560 cm(-1) (beta-sheet) appeared in pH 5.5-8.5 buffer solutions. A new broad shoulder at 1651 cm(-1) (random coil and/or alpha helix) with 1614 cm(-1) was observed in the pH 4.5 buffer solution. However, the broad shoulder at 1651 cm(-1) was converted to a maximum peak at 1679 cm(-1) (beta-turn/antiparallel beta-sheet) when the pH shifted from 4.5 to 3.5, but the original pronounced peak at 1614 cm(-1) became a shoulder. Once the pH was lowered to 2.5, the IR spectrum of PGAIPG was dominated by major absorption at 1679 cm(-1) with a minor peak at 1552 cm(-1) (alpha-helix/random coil). The result indicates that the pH was a predominant factor to transform PGAIPG structure from aggregated beta-sheet (pH 8.5) to beta-turn/intermolecular antiparallel beta-sheet (pH 2.5). Moreover, a partial conformation of PGAIPG with minor alpha-helix/random coil structures was also explored in the lower pH buffer solution. There was no thermal-dependent structural change for solid-state PGAIPG. The thermal-induced formation of aggregated beta-sheet for PGAIPG in aqueous solution was found from 28 to 30 degrees C, however, which might be correlated with the formation of an opaque gel that turned from clear solution. The formation of aggregated beta-sheet structure for PGAIPG beyond 30 degrees C might be due to the intermolecular hydrogen bonded interaction between the hydrophobic PGAIPG fragments induced by coacervation. PMID- 15752568 TI - 3D molecular modeling, free radical modulating and immune cells signaling activities of the novel peptidomimetic L-glutamyl-histamine: possible immunostimulating role. AB - An original representative of the patented by author family of histamine containing peptidomimetics L-glutamyl-histamine (L-Glu-Hist) was synthesized and characterized as a biologically active compound with a role of cytokine mimic leading to cellular responses of improved specificity. The study assesses the ability of L-Glu-Hist to affect molecular modeling, modulate free radical activity and influence immune cell signaling. The energy-minimized 3D conformations of L-Glu-Hist derived from its chemical structure resulted in stabilization for Fe2+ chelating complexes. L-Glu-Hist accelerated the decrease of ferrous iron in the ferrous sulfate solution in a concentration-dependent mode and showed the ferroxidase-like activity at concentrations less than 3 mM in the phenanthroline assay, whereas in the concentration range 3-20 mM L-Glu-Hist restricted the availability of Fe2+ to phenanthroline due to binding of ferrous ions in chelating complexes. L-Glu-Hist showed stimulatory effect on phosphatidylcholine liposomal peroxidation (LPO) catalyzed by the superoxide anion radical (O2*-)-generating system (Fe2+ + ascorbate) at low (less or about 1 mM) L-Glu-Hist concentrations and both revealed the inhibitory effect on LPO in this system of high (approximately 10 mM) L-Glu-Hist concentration. The stimulation of LPO by L-Glu-Hist was related to the ability of peptidomimetic in small (approximately 0.05 mM) concentrations to release O2*- free radicals as determined by the superoxide dismutase-inhibitable cytochrome c reduction assay. O2*- release by L-Glu-Hist might result from its ferroxidase-like activity, while inhibition of LPO by L-Glu-Hist was caused by its chelating activity to Fe2+ ions, prevention of free radical generation and lipid hydroperoxide-degrading ability of 5-20 mM L-Glu-Hist. L-Glu-Hist released O2*- in concentrations which stimulated [3H]-thymidine incorporation into DNA and proliferation of mouse spleen lymphocytes and mononuclear cells from human blood. L-Glu-Hist modulates the ability of oxygen free radicals to act as signaling agents at low concentrations, influencing gene expression. The structural peptide-like analogues of L-Glu-Hist such as L-Glu-Trp, carcinine (beta-alanylhistamine), but not L-Pro-Glu-Trp were active in stimulating thymidine incorporation and in inducing proliferation of mononuclear cells as compared to mitogen concanavalin A at doses 2.5-25.0 microg/ml. Our data provide evidence that L-Glu-Hist may act as a very fast, specific and sensitive trigger for lymphocyte proliferation and immunoregulation. The cited abilities and further obtained in vivo results make Immudilin ((INCI: glutamylamidoethyl imidazole, aqueous solution), L-Glu-Hist) a useful immunoregulatory agent. PMID- 15752569 TI - Phylloseptins: a novel class of anti-bacterial and anti-protozoan peptides from the Phyllomedusa genus. AB - Six novel peptides called phylloseptins (PS-1, -2, -3, -4, -5, and -6) showing anti-bacterial (PS-1) and anti-protozoan (PS-4 and -5) activities were isolated from the skin secretion of the Brazilian tree-frogs, Phyllomedusa hypochondrialis and Phyllomedusa oreades. Phylloseptins have a primary structure consisting of 19 21 amino acid residues (1.7-2.1 kDa). They have common structural features, such as a highly conserved N-terminal region and C-terminal amidation. Phylloseptin-1 (FLSLIPHAINAVSAIAKHN-NH2) demonstrated a strong effect against gram-positive and gram-negative bacteria (MICs ranging from 3 to 7.9 microM), without showing significant hemolytic activity (<0.6% at the MIC range) towards mammalian cells. Atomic force microscopy experiments indicated that the bacteriolytic properties of these peptides might be related to their disruptive action on the cell membrane, characterized by a number of bubble-like formations, preceding every cell lysis. PS-4 and PS-5 showed anti-protozoan activity with IC50 at about 5 microM for Trypanosoma cruzi. PMID- 15752570 TI - Isolation of trichogin, an antifungal protein from fresh fruiting bodies of the edible mushroom Tricholoma giganteum. AB - An antifungal protein was isolated from the mushroom Tricholoma giganteum var. golden blessings. The protocol included ion exchange chromatography on DEAE cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose, and gel filtration by fast protein liquid chromatography on Superdex 75. The antifungal protein, designated trichogin, was unadsorbed on DEAE-cellulose but was adsorbed on Affi-gel blue gel and CM cellulose. It exhibited antifungal activity against Fusarium oxysporum, Mycosphaerella arachidicola and Physalospora piricola. Trichogin inhibited HIV-1 reverse transcriptase with an IC50 of 83 nM. PMID- 15752571 TI - Kenojeinin I, antimicrobial peptide isolated from the skin of the fermented skate, Raja kenojei. AB - An antimicrobial peptide was purified from fermented skate skin extract using the solid-phase extraction and separation on HPLC reversed-phase chromatography. Amino acid sequence of the purified peptide (Peak A) having an antimicrobial activity revealed the presence of many cationic residues of the total 28 amino acids. Its molecular mass was found to be 3059 Da. This result was in excellent agreement with the theoretical molecular mass calculated from the amino acid sequence. The synthetic kenojeinin I had inhibitory effects on B. subtilis (MIC, 12 microg/ml), E. coli (28 microg/ml), and S. cerevisiae (12 microg/ml). These results indicate that fermented skate skin is potentially antimicrobial. PMID- 15752572 TI - Aplysia capsulin is localized to egg capsules and egg cordon sheaths and shares sequence homology with Drosophila dec-1 gene products. AB - Differential library screening of an albumen gland cDNA library, Western blot analysis, protein expression, immunolocalization studies, comparative genomics, and secretion assays identified a major Aplysia californica albumen gland protein ('capsulin') that is localized to egg capsules and to the sheaths of the egg cordon. Capsulin shared sequence homology with eggshell proteins encoded by the Drosophila dec-1 gene. The 1790-amino acid A. californica precursor contains 17 repeat sequences that are flanked by basic residue processing sites. The numerous proteolytic processing sites may facilitate the breakdown of capsulin prior to when veliger larvae break out of egg capsules as free-swimming larvae. An Aplysia brasiliana capsulin repeat sequence was 97% identical to its A. californica homolog. Capsulin fragments were not detected in the eluates of egg cordons, suggesting that capsulin is not a candidate water-borne pheromone precursor. PMID- 15752573 TI - Characterization of a peptide from skin secretions of male specimens of the frog, Leptodactylus fallax that stimulates aggression in male frogs. AB - During the breeding season of the mountain chicken frog Leptodactylus fallax, fighting between males results in the emergence of dominant animals that subsequently attract females to nesting sites. A peptide, termed Leptodactylus aggression-stimulating peptide (LASP), was isolated from norepinephrine stimulated skin secretions from male specimens of L. fallax that was not present in skin secretions obtained from females. The primary structure of the peptide was established as: Gly-Leu-Trp-Asp-Asp-Leu-Lys-Ala-Ala-Ala-Lys-Lys-Val-Val-Ser Ser-Leu-Ala-Ser-Ala-Ala-Ile-Glu-Lys-Leu NH2. LASP had no pheromone-like action on females but had a chemoattractive effect on males and stimulated aggressive behaviors, such as rearing and leaping. It is suggested that this peptide may play an important role in initiating the competitive male-male interactions that are associated with the onset of reproductive behavior in L. fallax. PMID- 15752574 TI - An opioid peptide from synganglia of the tick, Amblyomma testindinarium. AB - An opioid peptide, which shares similarity with mammalian hemorphins, has been identified from the synganglia (central nervous system) of the hard tick, Amblyomma testindiarium. Its primary sequence was established as LVVYPWTKM that contains a tetrapeptide sequence Tyr-Pro-Trp-Thr of hemorphin-like opioid peptides. By hot-plate bioassay, the purified peptide and synthetic peptide displayed dose-related antinociceptive effect in mice, as observed for other hemorphin-like opioid peptides. This is the first opioid peptide identified from ticks. Ticks may utilize the opioid peptide in their strategy to escape host immuno-surveillance as well as in inhibiting responses directed against themselves. PMID- 15752575 TI - Cardiovascular effects of endomorphins in alloxan-induced diabetic rats. AB - Endomorphins, the endogenous, potent and selective mu-opioid receptor agonists, have been shown to decrease systemic arterial pressure (SAP) in rats. In the present study, responses to endomorphins were investigated in systemic vascular bed of alloxan-induced diabetic rats and in non-diabetic rats. Diabetes was induced by alloxan (220 mg/kg, i.p.) in male Wistar rats. At 4-5 weeks after the onset of diabetes, intravenous injections of endomorphins (1-30 nmol/kg) led to an increase of SAP and heart rate (HR) consistently and dosed-dependently. SAP increased 7.68+/-3.73, 11.19+/-4.55, 21.19+/-2.94 and 27.48+/-6.21% from the baseline at the 1, 3, 10 and 30 nmol/kg dose, respectively, of endomorphin 1 (n=4; p<0.05), and similar changes were observed in response to endomorphin 2. The hypertension could be antagonized markedly by i.p. 2 mg/kg of naloxone. On the other hand, bilateral vagotomy would attenuate the effects of hypertension and diminished the changes of HR in response to endomorphins. With diabetic rats, 6-10 weeks after the induction of diabetes, intravenous injections of endomorphins produced non-dose-related various changes in SAP, such as a single decrease, or a single increase, or biphasic changes characterized by an initial decrease followed by a secondary increase, or no change at all. These results suggest that diabetes may lead to the dysfunction of the cardiovascular system in response to endomorphins. Furthermore, the diabetic rats of 4-5 weeks after alloxan-treatment, the increase in SAP and HR caused by i.v. endomorphins might be explained by a changed effect of vagus and by a naloxone-sensitive mechanism. PMID- 15752576 TI - Antinociceptive and antipyretic effects of a derivatized tetrapeptide from lactoferrin in rats. AB - PEP1261, a tetrapeptide derivative used in this study, corresponds to residues 39 42 of human lactoferrin. The parent protein lactoferrin is known to exhibit antinociceptive activity and it regulates many aspects of inflammation. This study is aimed to evaluate the antinociceptive and antipyretic activities of PEP1261 in rats. PEP1261 exhibits a significant dose dependent antinociceptive activity with optimal effect at 40 mg/kg body weight (b.w.) (i.p.) in both tail flick model and acetic acid induced writhing in rats. PEP1261 at the doses of 20 and 40 mg/kg b.w. (i.p.) is also observed to exhibit notable antipyretic effect in lipopolysaccharide-induced pyrexia in rats. In conclusion, the results suggest that PEP1261 possesses antinociceptive and antipyretic activities better than the control peptide KRDS. PMID- 15752577 TI - Reciprocal opioid-opioid interactions between the ventral tegmental area and nucleus accumbens regions in mediating mu agonist-induced feeding in rats. AB - Feeding elicited by the mu-selective agonist, [D-Ala2, M-Phe4, Gly-ol5] encephalin administered into the nucleus accumbens is blocked by accumbal pre treatment with mu, delta1, delta2 and kappa, but not mu1 opioid antagonists. Correspondingly, mu-agonist-induced feeding elicited from the ventral tegmental area is blocked by ventral tegmental area pre-treatment with mu and kappa, but not delta opioid antagonists. A bi-directional opioid-opioid feeding interaction has been firmly established such that mu-agonist-induced feeding elicited from the ventral tegmental area is blocked by accumbal naltrexone, and that accumbal mu-agonist-induced feeding is blocked by naltrexone pre-treatment in the ventral tegmental area. To determine which opioid receptor subtypes mediate the regional bi-directional opioid-opioid feeding interactions between these two sites, the present study examined the dose-dependent ability of either general (naltrexone), mu (beta-funaltrexamine), kappa (nor-binaltorphamine) or delta (naltrindole) opioid antagonists administered into one site to block mu-agonist-induced feeding elicited from the other site. General, mu and kappa, but not delta opioid receptor antagonist pre-treatment in the ventral tegmental area dose-dependently reduced mu-agonist-induced feeding elicited from the nucleus accumbens. General, mu and delta, and to a lesser degree kappa, opioid receptor antagonist pre treatment in the nucleus accumbens dose-dependently reduced mu-agonist-induced feeding elicited from the ventral tegmental area. Thus, multiple, but different opioid receptor subtypes are involved in mediating opioid-opioid feeding interactions between the nucleus accumbens and ventral tegmental area regions. PMID- 15752578 TI - High Fos expression during the active phase in orexin neurons of a diurnal rodent, Tamias sibiricus barberi. AB - To investigate whether a diurnal animal possesses the orexinergic system implicating vigilance and behavior, we examined Fos immunoreactivity (IR) in orexinergic neurons of Korean chipmunks raised under 12h light-dark cycles. Brain tissue, collected at four different zeitgeber times (ZT), was double-labeled with Fos and orexin-A antibodies. There was no difference in the number of orexin-IR neurons in the hypothalamus across all ZTs. However, more orexin-IR neurons expressing Fos-IR were found at ZTs 3 and 9 than ZTs 15 and 21. The results demonstrate circadian variations in the activation of orexin neurons corresponding with locomotor cycles, similarly seen in nocturnal rodents. PMID- 15752579 TI - CRF receptor type 1 mediates continual hypoxia-induced CRF peptide and CRF mRNA expression increase in hypothalamic PVN of rats. AB - We demonstrated previously that hypoxia activated CRF and CRF mRNA in PVN, and CRF receptor 1 (CRFR1) mRNA in rat pituitary. The aim of the study is to test whether the hypoxia-activated CRF and CRF mRNA is associated with triggering CRFR1. Rats were exposed to hypobaric hypoxia at altitude of 2 and 5 km. CRF and CRF mRNA were assayed by immunostaining and in situ hybridization. CRFR1 mRNA was assayed by RT-PCR. Results showed that 5 km continual hypoxia increased CRF and CRF mRNA in PVN, CRFR1 mRNA in pituitary, and plasma corticosterone. The hypoxia increased CRF, CRF mRNA, CRFR1 mRNA, and corticosterone were blocked by CRFR1 antagonist (CP-154,526), suggesting that CRFR1 in PVN and pituitary are responsible for the hypoxia-increased CRF and CRF mRNA in PVN. PMID- 15752580 TI - A comparison of leptin and ghrelin levels in plasma and saliva of young healthy subjects. AB - In the last 10 years, saliva has been increasingly used as a diagnostic fluid and in predictions of disease progression. Leptin and ghrelin are synthesized in several tissues including the salivary glands. The action of ghrelin is antagonistic to that of leptin. This study was undertaken to measure and compare the saliva ghrelin-leptin and plasma ghrelin-leptin levels in healthy young subjects. In 30 healthy subjects, after an overnight fast, saliva and plasma leptin levels were measured using the ELISA method while saliva and plasma immunoreactive ghrelin levels were measured using a commercial radioimmunoassay (RIA). The latter uses 125I-labeled bioactive ghrelin as a tracer and a rabbit polyclonal antibody raised against full-length octanoylated human ghrelin (Phoenix, Europe, Karlsruhe, Germany). The results of this investigation revealed that saliva leptin levels (6.19+/-2.10 microg/l) were lower than plasma levels (7.39+/-3.23 microg/l) while saliva ghrelin levels (188.5+/-84.7 pg/ml) were higher than plasma levels (126.4+/-38.5 pg/ml), when male and female subjects were considered together. Saliva leptin levels (5.93+/-1.94 microg/l) were lower than plasma levels (6.22+/-2.92 pg/ml) while saliva ghrelin levels (190.3+/-80.2 pg/ml) were higher than plasma levels (120.4+/-35.7 pg/ml) in young males. Saliva leptin levels (6.47+/-2.29 microg/l) were lower than plasma levels (8.73+/-3.14 microg/l) while saliva ghrelin levels (183.2+/-90.2 pg/ml) were higher than plasma levels (129.3+/-42.8 pg/ml) in young females, and both saliva and plasma leptin levels were slightly lower in male subjects in comparison with female subjects. Also, Immunohistochemistry study indicated that ghrelin positivity was found in ductus epithelium of salivary gland. We have demonstrated for the first time that saliva ghrelin levels were higher than in plasma while saliva leptin levels were almost the same as in plasma. Measurements of ghrelin and leptin in saliva is non-invasive, simple, and generally much preferred by patients and thus may be an acceptable alternative to plasma sampling. PMID- 15752581 TI - Central vagal stimulation activates enteric cholinergic neurons in the stomach and VIP neurons in the duodenum in conscious rats. AB - The influence of central vagal stimulation induced by 2h cold exposure or intracisternal injection of thyrotropin-releasing hormone (TRH) analog, RX-77368, on gastro-duodenal enteric cholinergic neuronal activity was assessed in conscious rats with Fos and peripheral choline acetyltransferase (pChAT) immunoreactivity (IR). pChAT-IR was detected in 68%, 70% and 73% of corpus, antrum and duodenum submucosal neurons, respectively, and in 65% of gastric and 46% of duodenal myenteric neurons. Cold and RX-77368 induced Fos-IR in over 90% of gastric submucosal and myenteric neurons, while in duodenum only 25-27% of submucosal and 50-51% myenteric duodenal neurons were Fos positive. In the stomach, cold induced Fos-IR in 93% of submucosal and 97% of myenteric pChAT-IR neurons, while in the duodenum only 7% submucosal and 5% myenteric pChAT-IR neurons were Fos positive. In the duodenum, cold induced Fos in 91% of submucosal and 99% of myenteric VIP-IR neurons. RX-77368 induces similar percentages of Fos/pChAT-IR and Fos/VIP-IR neurons. These results indicate that increased central vagal outflow activates cholinergic neurons in the stomach while in the duodenum, VIP neurons are preferentially stimulated. PMID- 15752582 TI - Neurotoxicity and oxidative stress in D1M-substituted Alzheimer's A beta(1-42): relevance to N-terminal methionine chemistry in small model peptides. AB - Small model peptides containing N-terminal methionine are reported to form sulfur centered-free radicals that are stabilized by the terminal N atom. To test whether a similar chemistry would apply to a disease-relevant longer peptide, Alzheimer's disease (AD)-associated amyloid beta-peptide 1-42 was employed. Methionine at residue 35 of this 42-mer has been shown to be a key amino acid residue involved in amyloid beta-peptide 1-42 [A beta1-42]-mediated toxicity and therefore, the pathogenesis of AD. Previous studies have shown that mutation of the methionine residue to norleucine abrogates the oxidative stress and neurotoxic properties of A beta(1-42). In the current study, we examined if the position of methionine at residue 35 is a criterion for toxicity. In doing so, we tested the effects of moving methionine to the N-terminus of the peptide in a synthetic peptide, A beta(1-42)D1M, in which methionine was substituted for aspartic acid at the N-terminus of the peptide and all subsequent residues from D1 to L34 were shifted one position towards the carboxy-terminus. A beta(1-42)D1M exhibited oxidative stress and neurotoxicity properties similar to those of the native peptide, A beta(1-42), all of which are inhibited by the free radical scavenger Vitamin E, suggesting that reactive oxygen species may play a role in the A beta-mediated toxicity. Additionally, substitution of methionine at the N terminus by norleucine, A beta(1-42)D1Nle, completely abrogated the oxidative stress and neurotoxicity associated with the A beta(1-42)D1M peptide. The results of this study validate the chemistry reported for short peptides with N-terminal methionines in a disease-relevant peptide. PMID- 15752583 TI - Characterization of a naturally-occurring polymorphism in the UHR-1 gene encoding the putative rat prolactin-releasing peptide receptor. AB - The rat orphan receptor UHR-1 and its human orthologue, GPR10, were first isolated in 1995. The ligand for this receptor, prolactin-releasing peptide (PrRP), was identified in 1998 by reverse pharmacology and has subsequently been implicated in a number of physiological processes. As supported by its localization and regulation in the hypothalamus and brainstem, we have shown previously that PrRP is involved in energy homeostasis. Here we describe a naturally occurring polymorphism in the UHR-1 gene that results in an ATG to ATA change at the putative translational initiation site. The presence of the polymorphism abolished the binding of 125I PrRP in rat brain slices but did not affect the ability of PrRP to reduce fast-induced food intake. Together this data suggest that PrRP may be exerting its feeding effects through a receptor other than UHR-1. PMID- 15752584 TI - Characterization of functional urotensin II receptors in human skeletal muscle myoblasts: comparison with angiotensin II receptors. AB - The properties of urotensin II (U-II) receptor (UT receptor) and angiotensin II (ANG II) receptor (AT receptor) in primary human skeletal myoblasts (HSMM) and differentiated skeletal myotubes (HSMMT) were characterized. Radiolabeled U-II and ANG II bound specifically to HSMM with Kd's of 0.31 nM (2311 receptors/cell) and 0.61 nM (18,257 receptors/cell), respectively. The cyclic segment of U-II peptide, CFWKYC, was the minimal sequence required for binding, with the WKY residues essential. Inhibitor studies suggested AT1 is the predominant ANG II receptor. After radioligand binding, under conditions designed to minimize receptor internalization, half the bound U-II was resistant to acid washing suggesting that U-II binds tightly to its receptor in a quasi-irreversible fashion. The AT1 receptor-bound radioligand was completely removed under the same conditions. RT-PCR detected the expression of mRNAs for UT and AT1 receptors. Western blotting showed that U-II and ANG II signaled via ERK1/2 kinase. UT receptor was not lost upon differentiation into myotubes since both mRNA for UT receptor and U-II binding were still present. ANG II receptors were also present as shown by ANG II-induced calcium mobilization. PMID- 15752585 TI - Distribution of atrial natriuretic peptide and its effects on contraction and intracellular calcium in ventricular myocytes from streptozotocin-induced diabetic rat. AB - The distribution of atrial natriuretic peptide (ANP) in blood plasma and cardiac muscle and its effects on ventricular myocyte contraction and intracellular free calcium concentration [Ca2+]i in the streptozotocin (STZ)-induced diabetic rat have been investigated. Blood plasma concentration and heart atrial and ventricular contents of ANP were significantly increased in STZ-treated rats compared to age-matched controls. STZ treatment increased the number of ventricular myocytes immunolabeled with antibodies against ANP. In control myocytes the percentage of cells that labeled positively and negatively were 17% versus 83%, respectively. However, in myocytes from STZ-treated rat the percentages were 52% versus 53%. Time to peak (TPK) shortening was significantly and characteristically prolonged in myocytes from STZ-treated rats (360+/-5 ms) compared to controls (305+/-5 ms). Amplitude of the Ca2+ transient was significantly increased in myocytes from STZ-treated rats compared to controls (0.39+/-0.02 versus 0.29+/-0.02 fura-2 RU in controls) and treatment with ANP reduced the amplitude of the Ca2+ transient to control levels. ANP may have a protective role in STZ-induced diabetic rat heart. PMID- 15752586 TI - Deficit in beta-endorphin peptide and tendency to alcohol abuse. AB - Human and animal studies suggest that there is a correlation between endogenous opioid peptides, especially beta-endorphin, and alcohol abuse. It has been proven that the consumption of alcohol activates the endogenous opioid system. Consumption of alcohol results in an increase in beta-endorphin level in those regions of the human brain, which are associated with a reward system. However, it has also been observed that habitual alcohol consumption leads to a beta endorphin deficiency. It is a well-documented phenomenon that people with a genetic deficit of beta-endorphin peptide are particularly susceptible to alcoholism. The plasma level of beta-endorphin in subjects genetically at high risk of excessive alcohol consumption shows lower basal activity of this peptide. Its release increases significantly after alcohol consumption. Clinical and laboratory studies confirm that certain genetically determined factors might increase the individual's vulnerability to alcohol abuse. PMID- 15752587 TI - Biology after the genome: a physical review. Proceedings of BIFI 2004 International Conference. February 11-13, 2004. Zaragoza, Spain. PMID- 15752588 TI - Heat capacity effects in protein folding and ligand binding: a re-evaluation of the role of water in biomolecular thermodynamics. AB - Large "anomalous" heat capacity (DeltaC(p)) effects are a common feature of the thermodynamics of biomolecular interactions in aqueous solution and, as a result of the improved facility for direct calorimetric measurements, there is a growing body of experimental data for such effects in protein folding, protein-protein and protein-ligand interactions. Conventionally such heat capacity effects have been ascribed to hydrophobic interactions, and there are some remarkably convincing demonstrations of the usefulness of this concept. Nonetheless, there is also increasing evidence that hydrophobic interactions are not the only possible source of such effects. Here we re-evaluate the possible contributions of other interactions to the heat capacity changes to be expected for cooperative biomolecular folding and binding processes, with particular reference to the role of hydrogen bonding and solvent water interactions. Simple models based on the hydrogen-bonding propensity of water as a function of temperature give quantitative estimates of DeltaC(p) that compare well with experimental observations for both protein folding and ligand binding. The thermodynamic contribution of bound waters in protein complexes is also estimated. The prediction from simple lattice models is that trapping of water in a complex should give more exothermic binding (DeltaDeltaH-6 to -12 kJ mol(-1)) with lower entropy (DeltaDeltaS(0) approximately -11 J mol(-1) K(-1)) and more negative DeltaC(p) (by about -75 J mol(-1) K(-1)) per water molecule. More generally, it is clear that significant DeltaC(p) effects are to be expected for any macromolecular process involving a multiplicity of cooperative weak interactions of whatever kind. PMID- 15752589 TI - Phase transition in vector spin glasses. AB - We first give an experimental and theoretical introduction to spin glasses, and then discuss the nature of the phase transition in spin glasses with vector spins. Results of Monte Carlo simulations of the Heisenberg spin glass model in three dimensions are presented. A finite size scaling analysis of the correlation length of the spins and chiralities shows that there is a single, finite temperature transition at which both spins and chiralities order. PMID- 15752590 TI - The effect of charge-introduction mutations on E. coli thioredoxin stability. AB - Technological applications of proteins are often hampered by their low-stability and, consequently, the development of procedures for protein stabilization is of considerable biotechnological interest. Here, we use simple electrostatics to determine positions in E. coli thioredoxin at which mutations that introduce new charged residues are expected to lead to stability enhancement. We also obtain the corresponding mutants and characterize their stability using differential scanning calorimetry. The results are interpreted in terms of the accessibility in the native structure of the mutated residues and the potential effect of the mutations on the residual structure of the denatured state. PMID- 15752591 TI - Low T scaling in the binary 2d spin glass. AB - We investigate 2d Ising spin glasses with binary couplings via exact computations of the partition function on lattices with periodic boundary conditions. After introducing the physical issues, we sketch the algorithm to compute the partition function as a polynomial with integer coefficients. This technique is then exploited to obtain the thermodynamic properties of the spin glass. We find an anomalous low temperature scaling of the heat capacity c(v) approximately e( 2beta) and that hyperscaling holds. PMID- 15752592 TI - ITC in the post-genomic era...? Priceless. AB - The information available after decoding the genome of the human species and many others is opening the possibility of new approaches to target thousands of protein interactions critical for a continuously increasing list of genetic and infectious diseases and pathologies, and to understand complex regulatory pathways and interaction networks describing cell function and interrelation. There is a need for a reliable technique offering the capability of measuring accurately macromolecular interactions (e.g. protein/ligand, protein/protein, protein/nucleic acid) in the laboratory. Compared to other analytical techniques, isothermal titration calorimetry (ITC) exhibits some important advantages for characterizing intermolecular interactions and binding equilibria. ITC is suitable for characterizing both low affinity interactions (e.g. protein network regulation and natural ligands) and high affinity interactions (e.g. rational drug design). Considering the advanced technological level reached as well as the outstanding quality of the information accessible through this technique, ITC is expected to play a very prominent role in the next years in the areas of rational drug design and protein network regulation. PMID- 15752593 TI - Core deformations in protein families: a physical perspective. AB - An analysis is presented on how structural cores change shape within protein families, and whether or not there is a relationship between these structural changes and the vibrational modes that proteins experiment due to topological constraints. A set of 13 representative and well-populated protein families are studied. The evolutionary directions of deformation are obtained by applying a new multiple structural alignment technique to superimpose the structures and extract a conserved core, together with Principal Components Analysis (PCA) to extract the main deformation modes. A low-resolution Normal Mode Analysis (NMA) technique is used in parallel to study the properties of the mechanical core plasticity of the same proteins. We find that the evolutionary deformations span a low dimensional space. A statistically significant correspondence exists between these principal deformations and the vibrational modes accessible to a particular topology. We conclude that, to a significant extent, the structures of evolving proteins seem to respond to sequence changes by collective deformations along combinations of low-frequency modes. The findings have implications in structure prediction by homology modeling. PMID- 15752594 TI - Activated dynamics and timescale separation within the landscape paradigm: signature of complexity, diversity and glassiness. AB - The landscape paradigm has become a widespread picture within the realm of complex systems. Complex systems include a great variety of systems, ranging from glasses to biopolymers, which display a common dynamical behavior. Within this framework, the dynamics of a such a system can be envisioned as the search it performs on its (potential energy) landscape. This approach rests on the belief that the relaxation behavior depends only on generic features, irrespective of specific details and lies on the validity of a timescale separation scenario computationally corroborated but not properly validated yet form first principles. In this work we shall show that the prevalence of activated dynamics over other kinds of mechanisms determines the emergence of complex dynamical behavior. Thus, complexity and diversity are not intrinsic properties of a system but depend on the kind of exploration of the landscape. We shall focus mainly on an ample generic context (complex hierarchical systems which have been used as models of glasses, spin glasses and biopolymers) and a specific one (model glass formers). For the last case we shall be able to reveal (in mechanistic terms) the microscopic rationale for the occurrence of timescale separation. Furthermore, we shall explore the connections between these two up to now mostly unrelated contexts and the relation to a variational principle, and we shall reveal the conditions for the applicability of the landscape approach. PMID- 15752595 TI - Quantum magnets with anisotropic infinite range random interactions. AB - Using exact diagonalization techniques, we study the dynamical response of the anisotropic disordered Heisenberg model for systems of S=1/2 spins with infinite range random exchange interactions at temperature T=0. The model can be considered as a generalization, to the quantum case, of the well-known Sherrington-Kirkpatrick classical spin glass model. We also compute and study the behavior of the Edwards Anderson order parameter and energy per spin as the anisotropy evolves from the Ising to the Heisenberg limits. PMID- 15752597 TI - Topological properties of the energy landscape of small peptides. AB - The topology of the potential energy landscape (PEL) underlying the dynamics of a two dimensional off-lattice model for a heteropolymer is analyzed for different sequences of amino-acids. A statistical characterization of the metastable minima and first-order saddles of the PEL highlights structural differences in the landscape of good and bad folding sequences and provides insight on the chain dynamics during folding. PMID- 15752596 TI - Theoretical models of possible compact nucleosome structures. AB - Chromatin structure seems related to the DNA linker length. This paper presents a systematic search of the possible chromatin structure as a function of the linker lengths, starting from three different low-resolution molecular models of the nucleosome. Gay-Berne potential was used to evaluate the relative nucleosome packing energy. Results suggest that linker DNAs, which bridges and orientate nucleosomes, affect both the geometry and the rigidity of the global chromatin structure. PMID- 15752598 TI - Analysis of PIN1 WW domain through a simple statistical mechanics model. AB - We have applied a simple statistical mechanics Go-like model to the analysis of the PIN1 WW domain, resorting to mean field and Monte Carlo techniques to characterize its thermodynamics, and comparing the results with the wealth of available experimental data. PIN1 WW domain is a 39-residue protein fragment which folds on an antiparallel beta-sheet, thus representing an interesting model system to study the behavior of these secondary structure elements. Results show that the model correctly reproduces the two-state behavior of the protein, and also the trends of the experimental phi(T) values. Moreover, there is a good agreement between Monte Carlo results and the mean field ones, which can be obtained with a substantially smaller computational effort. PMID- 15752599 TI - Relevant distance between two different instances of the same potential energy in protein folding. AB - In the context of complex systems and, particularly, of protein folding, a physically meaningful distance is defined which allows to make useful statistical statements about the way in which energy differences are modified when two different instances of the same potential-energy function are used. When the two instances arise from the fact that different algorithms or different approximations are used, the distance herein defined may be used to evaluate the relative accuracy of the two methods. When the difference is due to a change in the free parameters of which the potential depends on, the distance can be used to quantify, in each region of parameter space, the robustness of the modeling to such a change and this, in turn, may be used to assess the significance of a parameters' fit. Both cases are illustrated with a practical example: the study of the Poisson-based solvation energy in the Trp-Cage protein (PDB code 1L2Y). PMID- 15752600 TI - Native geometry and the dynamics of protein folding. AB - In this paper, we investigate the role of native geometry on the kinetics of protein folding based on simple lattice models and Monte Carlo simulations. Results obtained within the scope of the Miyazawa-Jernigan indicate the existence of two dynamical folding regimes depending on the protein chain length. For chains larger than 80 amino acids, the folding performance is sensitive to the native state's conformation. Smaller chains, with less than 80 amino acids, fold via two-state kinetics and exhibit a significant correlation between the contact order parameter and the logarithmic folding times. In particular, chains with N=48 amino acids were found to belong to two broad classes of folding, characterized by different cooperativity, depending on the contact order parameter. Preliminary results based on the Go model show that the effect of long range contact interaction strength in the folding kinetics is largely dependent on the native state's geometry. PMID- 15752601 TI - Classification of amino acids induced by their associated matrices. AB - In this paper we carry out an analysis of different types of potential and substitution matrices for amino acids, oriented to give a classification of the latter. The cluster decomposition is obtained, in a fully unsupervised way, from the subdominant ultrametric associated to the distance between amino acids induced by the corresponding matrix. In the comparative study, by looking at the classifications obtained from diverse matrices, we can get information on how they account for the different chemical-physical properties of the amino acids. PMID- 15752602 TI - Statistics of infections with diversity in the pathogenicity. AB - The statistics of outbreaks in a model for the propagation of meningococcal disease is analyzed, taking into account the possibility that the population is fragmented into weakly connected patches. It is shown that, depending on the size of the sample studied, the ration between the variance and the average of infected cases can vary from unity (Poisson statistics) to epsilon(-1), where epsilon is the normalized infection rate. PMID- 15752603 TI - Phase transition in tensionless surfaces. AB - We study the critical behavior of the Laplacian roughening model, which describes the growth of tensionless surfaces. This type of growth phenomena is very important, for instance, in biological membranes and in molecular beam epitaxy. We summarize previous analytical and numerical results and point out their contradictions and differences, thus making clear the context of our work. Our contribution, achieved through large-scale numerical simulations, is the confirmation that the model exhibits a single continuous phase transition: the transition takes place between a continuum massless (i.e., with infinite correlation length) bilaplacian behavior and a massive propagator (finite correlation length). PMID- 15752604 TI - Folding simulations of small proteins. AB - Understanding how a protein folds is a long-standing challenge in modern science. We have used an optimized atomistic model (united-residue force field) to simulate folding of small proteins of various structures: HP-36 (alpha protein), protein A (beta), 1fsd (alpha+beta), and betanova (beta). Extensive Monte Carlo folding simulations (ten independent runs with 10(9) Monte Carlo steps at a temperature) starting from non-native conformations are carried out for each protein. In all cases, proteins fold into their native-like conformations at appropriate temperatures, and glassy transitions occur at low temperatures. To investigate early folding trajectories, 200 independent runs with 10(6) Monte Carlo steps are also performed at a fixed temperature for a protein. There are a variety of possible pathways during non-equilibrium early processes (fast process, approximately 10(4) Monte Carlo steps). Finally, these pathways converge to the point unique for each protein. The convergence point of the early folding pathways can be determined only by direct folding simulations. The free energy surface, an equilibrium thermodynamic property, dictates the rest of the folding (slow process, approximately 10(8) Monte Carlo steps). PMID- 15752605 TI - Understanding the structural characteristics of compstatin by conformational space annealing. AB - The structural characteristics of the 13-residue compstatin molecule are investigated using the conformational space annealing (CSA) method with CHARMM force field and the GBSA continuum solvent model. In order to sample conformations in the energy range of the minimized NMR structures, we have used the stopping criterion to the CSA search when a conformation whose energy is less than -490 kcal/mol is found. With this stopping criterion, a great variety of conformations are generated around experimentally known structures. Twenty independent CSA runs starting from random states find 1000 representative conformations in the energy landscape of the compstatin, which are classified into thirty-one structural families. The majority of the conformations (94.4%) are in the coil state. Other conformers containing a 3(10)-helix, a pi-helix, a beta-hairpin, and an alpha-helix are also found. PMID- 15752606 TI - Protein structure prediction based on fragment assembly and parameter optimization. AB - We propose a novel method for ab-initio prediction of protein tertiary structures based on the fragment assembly and global optimization. Fifteen residue long fragment libraries are constructed using the secondary structure prediction method PREDICT, and fragments in these libraries are assembled to generate full length chains of a query protein. Tertiary structures of 50 to 100 conformations are obtained by minimizing an energy function for proteins, using the conformational space annealing method that enables one to sample diverse low lying local minima of the energy. Then in order to enhance the performance of the prediction method, we optimize the linear parameters of the energy function, so that the native-like conformations become energetically more favorable than the non-native ones for proteins with known structures. We test the feasibility of the parameter optimization procedure by applying it to the training set consisting of three proteins: the 10-55 residue fragment of staphylococcal protein A (PDB ID 1bdd), a designed protein betanova, and 1fsd. PMID- 15752607 TI - Shannon information, LMC complexity and Renyi entropies: a straightforward approach. AB - The LMC complexity, an indicator of complexity based on a probabilistic description, is revisited. A straightforward approach allows us to establish the time evolution of this indicator in a near-equilibrium situation and gives us a new insight for interpreting the LMC complexity for a general non equilibrium system. Its relationship with the Renyi entropies is also explained. One of the advantages of this indicator is that its calculation does not require a considerable computational effort in many cases of physical and biological interest. PMID- 15752609 TI - On the robustness of complex heterogeneous gene expression networks. AB - We analyze a continuous gene expression model on the underlying topology of a complex heterogeneous network. Numerical simulations aimed at studying the chaotic and periodic dynamics of the model are performed. The results clearly indicate that there is a region in which the dynamical and structural complexity of the system avoid chaotic attractors. However, contrary to what has been reported for Random Boolean Networks, the chaotic phase cannot be completely suppressed, which has important bearings on network robustness and gene expression modeling. PMID- 15752608 TI - Towards a new interaction enzyme:coenzyme. AB - Ferredoxin-NADP(+) reductase catalyses NADP(+) reduction, being specific for NADP(+)/H. To understand coenzyme specificity determinants and coenzyme specificity reversion, mutations at the NADP(+)/H pyrophosphate binding and of the C-terminal regions have been simultaneously introduced in Anabaena FNR. The T155G/A160T/L263P/Y303S mutant was produced. The mutated enzyme presents similar k(cat) values for NADPH and NADH, around 2.5 times slower than that reported for WT FNR with NADPH. Its K(m) value for NADH decreased 20-fold with regard to WT FNR, whereas the K(m) for NADPH remains similar. The combined effect is a much higher catalytic efficiency for NAD(+)/H, with a minor decrease of that for NADP(+)/H. In the mutated enzyme, the specificity for NADPH versus NADH has been decreased from 67,500 times to only 12 times, being unable to discriminate between both coenzymes. Additionally, giving the role stated for the C-terminal Tyr in FNR, its role in the energetics of the FAD binding has been analysed. PMID- 15752610 TI - Structure and conformational stability of the enzyme I of Streptomyces coelicolor explored by FTIR and circular dichroism. AB - The bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS), formed by a cascade of several proteins, couples the translocation and phosphorylation of specific sugars across cell membranes. The structure and thermal stability of the first protein (enzyme I, EI) of the PTS in Streptomyces coelicolor is studied by using far-UV circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) at pH 7.0. The deconvolution of FTIR spectra indicates that the protein is mainly composed by a 35% of alpha-helical structure and 30% of beta-sheet. The thermal denaturation curves, as followed by both techniques, show only a midpoint at 330 K. This thermal denaturation behaviour is different to that observed in other members of the EI family. PMID- 15752611 TI - Dynamics of antibodies from cryo-electron tomography. AB - The issue of protein dynamics and its implications in the biological function of proteins are arousing greater and greater interest in molecular biology. In cryo electron tomography experiments one takes several snapshots of a given biological macromolecule. In principle, a large enough collection of snapshots may then be used to calculate its equilibrium configuration in terms of the experimentally accessible degrees of freedom, and hence estimate its potential energy. Consequently, one could analyze the biological functions of biomolecules by directly accessing their dynamics. In this work, we analyze the results of cryo electron tomography experiments on monoclonal murine IgG2a antibodies. With the aid of a novel software for image processing, we measure the equilibrium distribution of the angles which describe the configuration of the molecule. This helps us shed some critical light on recent results from X-ray crystallography. We then build a model of the antibody dynamics, which enables us to use the measured angular distribution in order to derive an explicit expression of the IgG potential energy. Finally, as a preliminary application of our results, we investigate the dynamical effects in the rate of formation of the antigen antibody encounter complex. In particular, we suggest that the dynamics of antibodies operates in the direction of decreasing anticooperativity of the two antigen binding arms. PMID- 15752612 TI - Skipping of exon 2 and exons 2 plus 3 of HMG-CoA lyase (HL) gene produces the loss of beta sheets 1 and 2 in the recently proposed (beta-alpha)8 TIM barrel model of HL. AB - HMG-CoA lyase (HL) deficiency is a rare autosomal recessive genetic disorder that affects ketogenesis and leucine catabolism. We report a new Spanish patient who bears the frequent nonsense mutation G109T (Mediterranean mutation). This mutation can produce aberrant splicing with three mRNA variants: one of the expected size, the second with deletion of exon 2, and the third with deletion of exons 2 and 3. Recently our group proposed a 3D model for human HL containing a (beta-alpha)(8) (TIM) barrel structure. We have studied the effect of the deletions of exon 2 and exons 2 plus 3 on the proposed HL model. Exon 2 skipping led to the loss of beta-sheet 1, and the skipping of exons 2 and 3 caused the disappearance of alpha helix 1 and beta-sheets 1 and 2.- PMID- 15752613 TI - The revelation of expressing region in the processed ceruloplasmin gene in human genome by biocomputational and biochemical methods. AB - ANNOTATION: Translation in all open reading frames (ORF) of human ceruloplasmin (Cp) pseudogene revealed the only translating sequence of 984 nucleotides. The amino acid sequence contains a signal peptide for mitochondrial protein import at N-terminus. The predicted protein without taking the signal peptide into consideration has 92% identity to the corresponding Cp fragment. It contains 20 amino acid substitutions, 8 of them are significant. There is His-X-His motif in the center of a molecule that is typical for copper containing oxidases. Potential copper-binding site appears as a result of the substitution P923H along human Cp sequence. Cp pseudogene transcription product was found in the cultured human cell lines HepG2 and HuTu 80 using RT-PCR strategy. Cp polypeptides with molecular weight of nearly 30 kDa were found in mitochondria of HuTu 80 cells. The possible biological role of mitochondrial Cp is under discussion. PMID- 15752615 TI - Novel knowledge-based potentials for ligand docking: variational approach to the old problem. AB - The variational approach of evaluation for knowledge-based potentials is considered for the first time. In this approach, the problem to derive knowledge based potentials is solved as the optimization task in the multiparametric model of atom types, reference states and interaction cutoff radii. Using analogy to liquid state theory we offered four new reference states and derived corresponding knowledge-based potentials. The cutoff radii and atom types are optimized to minimize averaged root-mean square deviations (RMSD) of the ligand docked positions regarding to the experimentally determined poses. The number of atom types is varied on the developed atom type tree with 6 root (C, N, O, S, P and the halogen type) and 49 apical atom types. We showed a pronounced effect of atom type choice on docking accuracy and proved that splitting of elements C, N and O of the periodic system up to the 18 optimal atom types essentially improves docking accuracy. PMID- 15752614 TI - A new interpolation formula for semiflexible polymers. AB - A new formula for the force vs. extension relation is derived from the discrete version of the so-called Worm-like chain model. This formula correctly fits some recent experimental data on polymer stretching. Moreover, we have compared our formula with a Monte Carlo simulation of a semiflexible polymer. PMID- 15752616 TI - Empirical parametrization of pK values for carboxylic acids in proteins using a genetic algorithm. AB - Considerable effort has been devoted to the development of theoretical electrostatic methods to predict the pK values of ionizable residues in proteins. However, predictions appear often to be still at the qualitative or semi quantitative level. We believe that, with the increasing number experimentally available pK values for proteins of known structure, an alternative approach becomes feasible: the empirical parametrization of the experimental protein pK database. Of course, in the long term, this empirical approach is no substitute for rigorous electrostatic analysis but, in the short term, it may prove to have useful predictive power and it may help to pinpoint the main structural determinants of pK values in proteins. Here we demonstrate the feasibility of the parametrization approach by fitting (using a genetic algorithm as fitting tool) the database for carboxylic acid pK values in proteins on the basis of an empirical equation that takes into account the two following kinds of effects: (1) long-range charge-charge interactions; (2) interactions of the given carboxylic acid group with its environment in the protein, which are described in terms of contributions from the different kind of atoms present in the protein (atomic contributions). PMID- 15752617 TI - Towards a new therapeutic target: Helicobacter pylori flavodoxin. AB - Helicobacter pylori flavodoxin is the electronic acceptor of the pyruvate oxidoreductase complex (POR) that catalyzes pyruvate oxidative decarboxilation. Inactivation of this metabolic route precludes bacterial survival. Because flavodoxin is not present in the human host, substances interfering electronic transport from POR might be well suited for eradication therapies against the bacterium. H. pylori flavodoxin presents a peculiar cofactor (FMN) binding site, compared to other known flavodoxins, where a conserved aromatic residue is replaced by alanine. A cavity thus appears under the cofactor that can be filled with small organic molecules. We have cloned H. pylori fldA gene, expressed the protein in Escherichia coli and characterized the purified flavodoxin. Thermal up shift assays of flavodoxin with different concentrations of benzylamine, as well as fluorescence titration experiments indicate benzylamine binds in the pocket near the FMN binding site. It seems thus that low affinity inhibitors of H. pylori flavodoxin can be easily found that, after improvement, may give rise to leads. PMID- 15752618 TI - Stiff polymer adsorption. Onset to pattern recognition. AB - We present the results of extensive off-lattice Monte-Carlo simulations of a stiff polymer chain adsorbing onto a sticky periodic stripe-like pattern of variable width. We have analyzed, in terms of the chain length and rigidity, the adsorption and the pattern recognition process as a function of the stripe width. We have seen that this process is twofold: (i) the chain adsorbs rather isotropically onto the surface at a characteristic temperature T(c) and (ii) a further reduction in the temperature is needed for the chain to reorganize and adjust to the specific pattern. Such polymer reorganization has been studied through the evaluation of the chain degree of stretching and asphericity. We have found an optimal stripe width that maximizes the stretching. We have introduced a criteria to estimate the characteristic temperature at which the pattern recognition takes place T(r)50%) lipopolysaccharides (LPS)-induced NO release from RAW264.7 cells at 50 microg/ml, with 18 having greater than 100% inhibition. At 200 microg/ml, 61 methanol extracts exhibited inhibitory activity (>50%), with 45 showing greater than 100% inhibition. In addition, the free radical scavenging effects of 6 methanolic extracts were found to be more than 50% for extract concentration of 0.5 mug/ml. The results indicate that the extracts contain active compounds that inhibit NO release and scavenge free radical. PMID- 15752631 TI - Vasodilator activity of the aqueous extract of Viscum album. AB - The aqueous extract of Viscum album leaves showed a significant coronary vasodilator activity on the Langendorff's isolated and perfused heart model. The data obtained suggest that the aqueous extract of V. album contains some biologically active principles that may act as inducers of the nitric oxide/soluble guanylate cyclase pathway. PMID- 15752632 TI - Antitumor activity of three benzopyrans isolated from Hypericum polyanthemum. AB - In the present study we have investigated the in vitro antitumor effects of three benzopyrans, 6-isobutyryl-5,7-dimethoxy-2,2-dimethylbenzopyran (1), 7-hydroxy-6 isobutyryl-5-methoxy-2,2-dimethylbenzopyran (2) and 5-hydroxy-6-isobutyryl-7 methoxy-2,2-dimethylbenzopyran (3) isolated from Hypericum polyanthemum. The three compounds tested demonstrated potent growth inhibitory activity at 40 microg/ml (<25% control growth) in the NCI-H460, HT-29 and U-373MG human cell lines. Determination of cell cycle distribution demonstrated that the antiproliferative effect of the three benzopyrans could be associated to alterations in the cell cycle phase distribution. Treatment with the IC50 of the three compounds induced an arrested in S phase. Only in cells treated with compound 3 did the percentage of sub-G1 population increase up to 9%, suggesting that this compound induced more cell death than the others. Consistent with sub G1 analysis, appreciable oligonucleosomal DNA fragmentation was only observed in the NCI-H460 cell line treated with compound 3. From these results it can be suggested that despite no differences among the cytotoxicity of the three compounds, it was observed that the mechanism of their antiproliferative effects appears to be different. PMID- 15752633 TI - Antioxidant activity, reducing power and total phenolic content of some lichen species. AB - The total antioxidant activity, total phenolic content and reducing power of methanol and water extracts of four lichen species, Bryoria fuscescens, Dermatocarpon intestiniformis, Peltigera rufescens and Pseudevernia furfuracea, were determined in vitro. Water and methanol extracts of P. rufescens showed the highest antioxidant activity. However, there was no correlation between antioxidant activity and total phenolic content of the extracts. Although the methanol extract of P. furfuracea had the highest total phenolic contents, it exhibited low antioxidant activity. In contrast, there was a strong correlation between reducing power and total antioxidant activity of the extracts. The highest reducing power was determined for the methanol extract of P. rufescens. PMID- 15752634 TI - Antioxidant activity of water extract of Scoparia dulcis. AB - An aqueous extract of Scoparia dulcis showed marked antioxidative activity in vitro, which supports the therapeutic effects claimed by traditional practitioners. PMID- 15752635 TI - Hepatoprotective activity of Adhatoda vasica aqueous leaf extract on D galactosamine-induced liver damage in rats. AB - Adhatoda vasica leaf showed significant hepatoprotective effect at doses of 50 100 mg/kg, p.o., on liver damage induced by D-galactosamine in rats. PMID- 15752636 TI - Antimicrobial activity of Sternbergia sicula and Sternbergia lutea. AB - n-Hexane, ethyl acetate, ethanolic and aqueous extracts of the bulbs and aerial parts of Sternbergia sicula and Sternbergia lutea were tested for their antimicrobial and antifungal activity. PMID- 15752637 TI - Antinociceptive diterpene from Euphorbia decipiens. AB - One myrsinol-type diterpene ester (1) isolated from Euphorbia decipiens was evaluated for analgesic activity in the acetic acid induced writhing test in mice. Different dose (5-20 mg/kg i.p.) of the compound showed significant antinociceptive activity, which was comparable to standard analgesic drugs, aspirin and ibuprofen (100 mg/kg i.p.). PMID- 15752638 TI - Antibacterial properties of essential oils from Thai medicinal plants. AB - By using disc diffusion assay, the antimicrobial activity of 32 essential oil samples extracted from local plants or plants cultivated in Thailand was evaluated against zoonotic enteropathogens including Salmonella spp., Escherichai coli O157, Campylobacter jejunii and Clostridium perferingens which are important for broiler export. Out of the essential oil tested, only the essential oil of Zingiber cassumuna, Cinnamomum bejolghota, Mentha arvensis var. piperacens, Cymbopogon citratus and Ocimum basilicum var. citratum showed promising antibacterial activity against the bacteria tested. PMID- 15752639 TI - Antibacterial activity of the endemic Hypericum kazdaghensis. AB - The antibacterial activity of the chloroform, acetone and methanol extracts of endemic Hypericum kazdaghensis leaves were evaluated by disc diffusion method. All extracts exhibited antimicrobial activity against all tested bacteria. PMID- 15752640 TI - Antibacterial activity of Eupatorium glandulosum leaves. AB - Antibacterial screening of petroleum ether, chloroform, ethyl acetate, methanol and aqueous extracts of Eupatorium glandulosum leaves exhibited a broad spectrum of inhibitory activity against Gram (+) and Gram (-) pathogenic bacteria. PMID- 15752641 TI - Antifungal activity of Rubia tinctorum, Rhamnus frangula and Caloplaca cerina. AB - The results of a preliminary antifungal screening of the methanol extracts and the major anthraquinone aglycones, alizarin and emodin, of Rubia tinctorum and Rhamnus frangula in comparison with the antifungal activity of the anthraquinone containing lichen Caloplaca cerina and its main secondary metabolite parietin are reported. PMID- 15752642 TI - Sensitivity of fungi isolated from onychomycosis to Eugenia cariophyllata essential oil and eugenol. AB - The antifungal activity of Eugenia cariophyllata essential oil and eugenol, its major constituent, on fungal strains isolated from onychomycosis was evaluated. The natural products presented prominent antifungal action with MIC of 1% and 4%, respectively. PMID- 15752643 TI - In vitro spasmolytic effect of aqueous extract of Calotropis procera on Guinea pig trachea smooth muscle chain. AB - The aqueous extract of Calotropis procera was evaluated for its spasmolytic effect using in vitro trachea smooth muscle chain of Guinea-pig. The extract (50, 100 and 200 microg/ml) showed a dose-dependent relaxant activity probably exhibited through the direct relaxant action on the smooth muscle. PMID- 15752644 TI - Antibacterial and antifungal activity of pinosylvin, a constituent of pine. AB - The antibacterial and antifungal activities of pinosylvin (3,5-dihydroxy-trans stilbene), a constituent of pine, were studied and compared with those of resveratrol (3,5,4'-trihydroxy-trans-stilbene). Pinosylvin exhibited more potent growth inhibitory activity against Candida albicans and Saccharomyces cerevisiae. PMID- 15752645 TI - Antinociceptive activity of Ceriops decandra leaf and pneumatophore. AB - The ethanol extract of Ceriops decandra leaf and pneumatophore, at the oral doses of 250 and 500 mg/kg, showed a dose-dependent and significant inhibition of acetic acid-induced writhing in mice. On the contrary, the bark extract was devoid of any significant activity. PMID- 15752646 TI - Antifungal activities of five Combretaceae used in Togolese traditional medicine. AB - Five species of Combretaceae growing in Togo were investigated for their antifungal activity against 20 pathogenic fungi (10 yeast and 10 filamentous fungi). The five hydroethanolic extracts of Terminalia glaucescens and Anogeissus leiocarpus appeared to be the most active, their MICs ranging from 0.25 mg/ml to 4 mg/ml. The results confirm the traditional therapeutic properties of these plants. PMID- 15752648 TI - A new alkaloid from the fruits of Celastrus orbiculatus. AB - From the fruits of Celastrus orbiculatus, a new alkaloid 3-oxo-4-benzyl-3, 4 dihydro-1H-pyrrolo [2, 1-c] oxazine-6-methylal (1), was isolated. The structure was elucidated on the basis of spectroscopic methods. PMID- 15752647 TI - Flavonoids of Helichrysum compactum and their antioxidant and antibacterial activity. AB - From the capitula of Helichrysum compactum, the flavonoids apigenin, kaempferol, luteolin, naringenin, 3,5-dihydroxy-6,7,8-trimethoxyflavone, kaempferol-3-O glucoside, luteolin-7-O-glucoside and luteolin-4',7-di-O-glucoside and from the leafy stems apigenin, kaempferol, luteolin, quercetin, apigenin-7-O-glucoside, luteolin-7-O-glucoside, and quercetin-3-O-glucoside were isolated. Extracts of the capitula of H. compactum show antioxidant activity by inhibition of lipid peroxidation and also show antibacterial activity. PMID- 15752649 TI - p,p'-DDE depresses the immune competence of chinook salmon (Oncorhynchus tshawytscha) leukocytes. AB - p,p'-DDE, the main metabolite of DDT, is still detected in aquatic environments throughout the world. Here, the effects and mechanisms by which p,p'-DDE exposure might affect the immune system of chinook salmon (Oncorhynchus tshawytscha) was studied. Isolated salmon splenic and pronephric leukocytes were incubated with different concentrations of p,p'-DDE, and cell viability, induction of apoptosis, and mitogenic responses were measured by flow cytometry and Alamar Blue assay. p,p'-DDE significantly reduced cell viability and proliferation and increased apoptosis. The effect of p,p'-DDE on pronephric leukocytes was more severe than on splenic leukocytes, likely because pronephric leukocytes had a higher proportion of granulocytes, cells that appear more sensitive to p,p'-DDE. The effect of p,p'-DDE on leukocytes appeared to vary between developmental stages or seasonal differences. The mitogenic response of leukocytes of chinook salmon exposed to p,p'-DDE in vivo exhibited a biphasic dose-response relationship. Only leukocytes isolated from salmon treated with 59 ppm p,p'-DDE had a significantly lower percentage of Ig+ blasting cells than controls, although the response was biphasic. These results support the theory that exposure to chemical contaminants could lead to an increase in disease susceptibility and mortality of fish due to immune suppression. PMID- 15752650 TI - Differential spontaneous killing of human and murine tumour cell lines by leucocyte subpopulations from carp peripheral blood leucocytes. AB - Cell populations from carp (Cyprinus carpio L.) peripheral blood leucocytes (PBLs) were examined for nonspecific cytotoxicities. By using monoclonal antibodies (MAbs) against carp thrombocytes (TCL-HB8) and both neutrophils and monocytes (TCL-BE8), PBLs with a density of 1.08 g ml-1 were separated into three fractions: thrombocytes, a mixture of neutrophils and monocytes, and other cells (mainly lymphocytes), and the separated cells were tested for cytotoxic activities against mammalian tumour cell lines (K562, HeLa, P815 and Yac-1 cell). Consequently, the mixture of neutrophils and monocytes exhibited cytolysis against these target cells, whereas the lymphocyte-rich and thrombocyte fractions did not show any cytolysis. To isolate only neutrophils, which do not contain monocytes, the MAb (TCL-BE8) positive cells from PBLs with a density of 1.08-1.09 g ml-1 were separated. Pure isolated neutrophils showed cytotoxic activities against K562 cells, but not P815 cells. Furthermore, analysis of the cytolytic mechanisms indicated that killing of these cells depended on H2O2 or HOCl. These results suggest that both neutrophils and monocytes are effectors for nonspecific cytotoxicity in carp PBLs, and neutrophils may be distinct from monocytes in their reactivity in cytolysis, including target cell selectivity and/or target cell sensitivity, and the cytolytic pathway. In carp, cytotoxicity of target cells can be mediated by several populations of their leucocytes which have cytotoxic capacities with various recognition and cytolytic mechanisms. PMID- 15752651 TI - The immunostimulatory effect of hot-water extract of Gracilaria tenuistipitata on the white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus. AB - The total haemocyte count (THC), phenoloxidase activity, respiratory burst, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency to the pathogen Vibrio alginolyticus were examined in the white shrimp Litopenaeus vannamei (10.3+/-1.5 g) injected individually with hot-water extract of Gracilaria tenuistipitata at 4 or 6 microg g-1. L. vannamei receiving hot water extract of G. tenuistipitata at either dose increased significantly its THC, phenoloxidase activity, and respiratory burst after 2 days. L. vannamei received hot-water extract of G. tenuistipitata at 6 microg g-1 increased its phagocytic activity and clearance efficiency to V. alginolyticus after 1 day. In another experiment, L. vannamei which had been injected with hot-water extract of G. tenuistipitata were challenged with V. alginolyticus at 2x10(6) colony-forming units (cfu) shrimp-1 and then placed in seawater of 34 per thousand. The survival of shrimp that received hot-water extract of G. tenuistipitata at 6 microg g-1 was significantly higher than that of shrimp that received saline and the control shrimp after 3 days, and at the termination of the experiment (6 days after the challenge). It is therefore concluded that L. vannamei receiving the hot-water extract of G. tenuistipitata at 6 microg g-1 or less increased its immune ability and resistance to V. alginolyticus infection. PMID- 15752652 TI - Purification and characterisation of phenoloxidase from amphioxus Branchiostoma belcheri tsingtauense. AB - Phenoloxidase (PO) from the humoral fluid of amphioxus B. belcheri tsingtauense was purified using a sequential combination of ammonium sulphate precipitation, Sephadex G-200 chromatography and DEAE Sepharose Fast Flow chromatography. In PAGE, the purified enzyme exhibited a single band of 150 kDa under non-reducing conditions, and was resolved to three bands with molecular masses of 72, 46 and 44 kDa, respectively, under reducing conditions, suggesting that the PO in amphioxus humoral fluid seems to be a heterotrimer of three polypeptides held together by disulphide bonds. The substrate specificity and inhibition characteristics both indicate that the PO isolated from amphioxus humoral fluid is a tyrosinase-type enzyme. In addition, mouse antisera against the purified PO were prepared, and their specificity was confirmed by Western blotting, facilitating the future determination of the origin of PO in the humoral fluid and the distribution of PO-synthesising tissues in amphioxus. PMID- 15752653 TI - On the structure of intestine and presence of endocytic cells in the lamina propria of platy and black tetra (Teleostei). AB - The structure of the intestine in platy (Xiphophorus maculatus) and black tetra (Gymnocorymbus ternetzi) and the capability of cells within the intestinal wall to endocytose intraperitoneally injected horse-spleen ferritin, are described. The intestinal epithelial layer has about the same thickness in both species, but the width of the lamina propria and tunica muscularis in black tetra was only about 1/5 of that in platy. Ferritin was taken up by numerous cells within the lamina propria throughout the entire length of the platy intestine. The uptake was demonstrated as large and strongly coloured intracellular Prussian blue granules in sections treated with acid ferrocyanide. There was no such uptake by the lamina propria in black tetras. We suggest that the high numbers of endocytic cells within the intestinal lamina propria of platies provide a local defence against foreign cells and particles. Such a functional role may to some extent compensate for the lack of an HCl-based defence in the digestive system of this stomach-less species. PMID- 15752655 TI - Identification and characterisation of a homolog of an activation gene for the recombination activating gene 1 (RAG 1) in amphioxus. AB - Expression of recombination activating genes (RAG) involved in the V (D) J recombination is regulated by the RAG1 gene activator (RGA) in mammals. The sequence of a cDNA clone from an amphioxus cDNA library was found to be homologous to that of RGA from mouse stromal cells with 45% identity. The full length cDNA sequence comprises 1119 bp and encodes a putative protein of 210 amino acid residues. Characterisation of the amino acid sequence revealed that two MtN3 domains and seven transmembrane spans are present in this protein, indicating a potential role as a plasma membrane protein. This gene is expressed in many tissues and at differential developmental stages. A high expression level of RGA is detected in gonad tissues, and gastrula embryo and adult stages. The presence of the RGA gene in amphioxus suggests that the signal pathway required for the expression of RAG could exist in this primitive protochordate. It also implies that in the related molecules, primitive adaptive immunity may have existed in cephalochordate although the complete machinery of VDJ rearrangement may not be formed. PMID- 15752654 TI - Evaluation of DNA vaccination of spotted sand bass (Paralabrax maculatofasciatus) with two major outer-membrane protein-encoding genes from Aeromonas veronii. AB - Genes encoding two major outer membrane proteins (OMPs) of the bacterial pathogen Aeromonas veronii, Omp38 and Omp48, were used to construct DNA vaccines. The protective effect of such vaccines against motile aeromonad septicaemia was evaluated in spotted sand bass (Paralabrax maculatofasciatus), an endemic species of the Mexican Northwest Pacific coast and a potential resource for the aquaculture industry. Weak protein expression, as determined by immunoblotting, was observed after transfection of eukaryotic cells with the DNA vaccines. Fish immunized with a single intramuscular injection of 20 microg of the omp38 and omp48 DNA vaccines showed slightly, but significantly elevated serum antibody levels 4 and 6 weeks after vaccination, compared to fish vaccinated with the control plasmid pcDNA3.1. Spotted sand bass vaccinated with the omp38 and omp48 DNA vaccines and challenged with A. veronii by intraperitoneal route recorded a relative percent survival (RPS) between 50 and 60%. Histopathological signs of motile aeromonad septicaemia were observed in around 40% of omp38 and omp48 vaccinated fish and 80% of pcDNA3.1-vaccinated control fish. The results indicate that P. maculatofasciatus vaccinated with a single dose of DNA plasmids encoding the major OMPs from A. veronii shows partial protection against infection and mortality by A. veronii experimental infection. PMID- 15752656 TI - Biological effect of vaccination can be assessed directly from diluted whole blood of rainbow trout using homologous blood phagocytes as immunosensors. AB - A rapid and simple method is presented for determining antibody activity following vaccination, directly from diluted fish blood. The proposed method evaluates the effects of specific antibodies on ingestion by blood phagocytes, and may be used for measuring antibody levels following vaccination. The enhancing effect of trout IgM on ingestion was measured by luminol-amplified chemiluminescence (CL) emission of blood phagocytes. Respiratory burst (RB) activity of blood phagocytes was induced with the strain MT004 of bacterial fish pathogen Aeromonas salmonicida. To determine the boosting level of specific IgM on ingestion, various volumes of purified trout IgM containing specific antibodies against A. salmonicida were added to blood samples collected from non vaccinated fish, and the RB activity of blood phagocytes was measured. The presence of antibodies in plasma of artificially prepared immune blood (AIB) was confirmed using enzyme-linked immunosorbent assay (ELISA). At a final blood dilution of 1:250, the mean RB activity of blood samples boosted with IgM was more than seven times higher, compared to other tested blood dilutions boosted with equal amount of IgM. Accordingly, a dilution of 1:250 was employed in the field study of vaccinated and non-vaccinated fish. The levels of A. salmonicida specific antibodies in plasma samples of vaccinated and non-vaccinated fish were additionally confirmed with the ELISA assay. Based on these results, it is proposed that the biological activity of elicited antibodies can be assessed directly from diluted fish blood, using homologous blood neutrophils as immune sensors. PMID- 15752657 TI - Molecular characterisation of a turbot Mx cDNA. PMID- 15752658 TI - [The Bioethics law]. PMID- 15752659 TI - [Leucocytospermia, oxidative stress and male fertility: facts and hypotheses]. AB - Leukocytospermia is frequent and significantly increased (over 10(6)/ml) in 20% of male factor infertility. It induces the production of highly toxic reactive oxygen species (ROS) which impair genital track accessory glands and sperm cell functions. The seminal medium contains extremely potent antioxidative defenses which usually balance the oxidative stress. In vivo, these defenses can be overwhelmed when ROS production is extremely important and/or when it lasts for a very long period of time. Infertility can then appear. In vitro, ROS have been univoqually demonstrated for being highly toxic since spermatozoa are no longer protected. Sperm cell defects are : decrease of acrosome reaction and fusiogenic ability and increase of DNA fragmentations. In case of male factor infertility, a leukocytospermia represents an essential or an additional risk factor that should be treated, specially when in vitro therapy is to be scheduled, in order to improve gamete quality. PMID- 15752660 TI - [Hysterectomies in patients with no history of vaginal delivery. A study of 243 cases]. AB - OBJECTIVE: The object of this study was to audit the policy of hysterectomy in nulliparous women in a university hospital. PATIENTS AND METHODS: A retrospective medical records analysis of all hysterectomies performed during an 8-year period. Patients with no history of vaginal delivery were stratified into three groups: group 1, patients who underwent abdominal hysterectomies; group 2, patients undergoing vaginal hysterectomy (2a) or laparoscopy-assisted vaginal hysterectomy (2b). The groups were compared as to demographic data, surgical complications and outcomes. RESULTS: During the study period, there were 243 hysterectomies in patients with no history of vaginal delivery. Among these, vaginal hysterectomies (group 2) were undertaken in 75% (182 patients) and successfully performed in all but 13 patients (7.1%). Mean uterine weight was 943 grams in group 1 and 370 grams in group 2. Abdominal route (group 1) was associated with longer operative time (average: 105 min) than vaginal route (group 2a; 81 min) but shorter operative time that laparoscopy-assisted vaginal route (group 2b; 173 min). There was no significant difference in mean estimated blood loss and complications rates between groups 1 and 2. Hospital stay was shorter in group 2. Laparoscopic assistance was not associated with bigger uteri, neither with fewer complications. DISCUSSION AND CONCLUSION: Nulliparity should no longer be considered a contraindication to vaginal hysterectomy. In such patients, many more hysterectomies should be carried out vaginally and laparoscopic assistance does not offer obvious advantages over the standard vaginal approach. PMID- 15752661 TI - [Comparison of pre-induction ultrasonographic cervical length and Bishop score in predicting risk of cesarean section after labor induction with prostaglandins]. AB - OBJECTIVE: To compare pre-induction ultrasonographic cervical length and Bishop score in predicting risk of caesarean section after labor induction with prostaglandins. PATIENTS AND METHODS: Assessment of the Bishop score and measurement of the cervical length by transvaginal sonography were performed by two operators, blinded to each other's results among women with singleton pregnancies at between 34(+0) - 41(+3) weeks of gestation requiring induction of labor with prostaglandins for medical indications. Fisher's exact test and regression logistic models were used for statistics analysis. In order to measure the strength of the association between ultrasonographic cervical length or Bishop score on one hand, and the caesarean sections rate (global or for failed induction or failure to progress) on the other hand, we computed odds ratios with 95% confidence interval. RESULTS: Among the 266 patients included in the study, multivariate analysis has shown that only Bishop score was predictive for the global caesarean section risk (OR [95% CI] 0.63 [0.45-0.87] ; P =0.005). However, neither Bishop score (OR [95% CI] 0.68 [0.42-1.09] ; P =0.11), nor ultrasonographic cervical length (OR [95% CI] 1.01 [0.95-1.08] ; P =0.59) was predictive for failed induction or failure to progress caesarean section risk. DISCUSSION AND CONCLUSION: The Bishop score appears to be a better predictor of the global caesarean section risk than ultrasonographic cervical length after induction of labor for medical reasons. PMID- 15752662 TI - [Pelvic and para-aortic lymphatic involvement in tubal carcinoma: topography and surgical implications]. AB - OBJECTIVE: The purpose of this study is to determine the topography of pelvic and para-aortic node involvement in Fallopian tube carcinoma (PFTC). This will help us to recommend appropriate surgical treatment options to the related patients. PATIENTS AND METHOD: A retrospective study was performed on 19 women with PFTC who underwent a systematic bilateral pelvic and para-aortic lymphadenectomy. RESULTS: The overall frequency of lymph node involvement was 47% (9/19). The frequency of pelvic and para-aortic metastases was 21% (4/19) and 42% (8/19) respectively. The frequency of lymph node metastases according to the stage of the disease (stage I, II and III) was : 29% (2/7), 50% (1/2) and 60% (6/10) respectively. The left para-aortic chain above the level of the inferior mesenteric artery was the site most frequently involved (75%) when para-aortic nodes were involved. DISCUSSION AND CONCLUSIONS: In patients with primary tubal carcinoma, the left para-aortic chain above the level of the inferior mesenteric artery is the most frequently involved. A complete lymphadenectomy (including all pelvic and para-aortic chains up to the level of the left renal vein) should be performed in patients with primary tubal carcinoma, even in patients with stage I disease. PMID- 15752663 TI - [The antigonadrotropic activity of chlormadinone acetate in reproductive women]. AB - INTRODUCTION: Progesterone modulates pulsatile secretion of LH throughout the normal menstrual cycle in women. This effect is observed essentially at hypothalamic level via progesterone receptors, with a decline in frequency of GnRH cyclic secretion. It was observed that when several nor-progesterone and nor testosterone derived progestins were administered, the antigonadotropic function of the hypothalamic-pituitary-ovarian axis was inhibited. OBJECTIVE: Determine chlormadinone acetate's antigonadotropic activity when administered according to two schemes: from the day 5 to day 25 (L5) and from day 8 to day 25 (L8) of the menstrual cycle. PATIENTS AND METHODS: Randomized monocentric study carried out in twenty normally cycling healthy women included in two parallel groups and receiving 10 mg of chlormadinone acetate once a day, either from day 5 to day 25, or from day 8 to day 25 of the menstrual cycle. Hormonal dosages were determined with blood samplings performed each day from day 5 to day 25 of each menstrual cycle for estradiol, FSH and LH and from day 14 to day 25 of each cycle for progesterone. RESULTS: Sixteen women in group L5 and 14 in group L8 were kept for per protocol analysis. At the first cycle of treatment a decline in LH peak amplitude was observed in all women (16/16) of the L5 group and in 12 women out of 14 in the L8 group. For all women under treatment, graphs show a decrease in progesterone secretion parallel to decline in LH secretory peak amplitude as well as significant decline in estradiol secretion. Clinical tolerance was good. DISCUSSION AND CONCLUSION: Chlormadinone acetate induces anti-gonadotropic activity which is similar to other nor-progesterone and nor-testosterone derived progestins. This activity is shown at first cycle when the drug is used from day 5; when the drug is used from day 8, cycle 2 or 3 must be waited for in order to obtain the same result. PMID- 15752664 TI - [Small cell carcinoma of the ovary of the hypercalcemic type revealed by a severe acute pancreatitis: about one case]. AB - Small cell carcinoma of the ovary of the hypercalcemic type is a rare tumour, usually lethal and occurring almost exclusively in young patients. In the majority of described cases, signs of this lesion were revealed by the associated hypercalcemia or by virtue of the physical tumour bulk alone. We report the first case of ovarian small cell carcinoma of the hypercalcemic revealed by a severe acute pancreatitis in a 19-year-old patient. PMID- 15752665 TI - [Risk taking behaviors: girls' and boys' specificities. An anthropological approach]. AB - Risk taking behaviors show attempts to escape from suffering, to struggle to live. They are different for boys and girls. Girls take suffering upon themselves and make of their bodies the very scene of that suffering, whereas boys throw themselves against the world in provocative, transgressive behaviours. Besides, when the will of the girl is to be unique, the will of the boy is to be the best. PMID- 15752666 TI - [The limits of laparoscopic myomectomy]. AB - Feasibility of laparoscopic myomectomy has been already shown with numerous clinical studies. Short-term benefits of this procedure are nowadays established. Its limits are related to the surgical technique, the myoma process, and the clinical context of the patient. By using preoperative exclusion criteria, particularly the size and the number of myomas, laparoscopic treatment is possible with little laparoconversion and complications rates. One could be in doubt about the risk of uterine rupture during a pregnancy occurring after laparoscopic myomectomy. In infertile patients, this procedure is as effective as laparotomy, even though its benefits in terms of postoperative adhesions should be demonstrated. Less invasive surgery should be preferred. PMID- 15752667 TI - [Advances in cervical pathology: the perspective of the European colposcopists]. PMID- 15752668 TI - [Initial or interval debulking surgery for advanced stage ovarian cancer: state of-the-art. How to select patients?]. AB - The management of advanced stage ovarian cancer has been deeply modified over the last few years. In patients with massive peritoneal spread, the use of neoadjuvant chemotherapy, followed by interval surgery, reduces the morbidity of radical surgery with an improvement of the quality of life. Nevertheless, results of ongoing randomized studies should be waited before stating about the results on survival of such management compared to initial debulking surgery. Waiting such results, the standard treatment of advanced stage ovarian cancer in 2005 remains initial surgery, performed in order to obtain ideally a total resection of all macroscopic diseases, and followed by adjuvant chemotherapy. However, in patients with massive spread, interval debulking surgery is becoming an interesting option, and will perhaps become a standard management. But criteria to select patients between initial and interval debulking surgery should be clearly defined. Those different points will be studied in this paper. PMID- 15752669 TI - [For or against transvaginal digital examination in obstetrics]. PMID- 15752670 TI - [For transvaginal digital examination in obstetrics]. PMID- 15752671 TI - [Against systematic transvaginal digital examination in obstetrics]. PMID- 15752673 TI - [Recommendations on the management of cerebral ventriculomegaly (January 2004)]. PMID- 15752674 TI - [Response of D. Dewailly to the article by S. Trimeche et al]. PMID- 15752675 TI - [Response of M. Cosson to the article by G. Bader et al]. PMID- 15752676 TI - European cystic fibrosis society consensus on standards--a roadmap to "best care". PMID- 15752677 TI - Standards of care for patients with cystic fibrosis: a European consensus. PMID- 15752678 TI - Docosahexaenoic acid trials in cystic fibrosis: a review of the rationale behind the clinical trials. PMID- 15752679 TI - Long-term azitromycin treatment of cystic fibrosis patients with chronic Pseudomonas aeruginosa infection; an observational cohort study. AB - BACKGROUND: In cystic fibrosis (CF), chronic endobronchial infection with Pseudomonas aeruginosa is a serious complication. Macrolides can increase lung function and weight in patients, and reduce exacerbations. METHODS: In 2001, we introduced long-term, low-dose azithromycin (AZ) treatment as an integral part of our routine treatment of these patients. Our study is an observational cohort study of all CF patients with chronic P. aeruginosa infection in our CF center comparing clinical parameters of the patients 12 months prior to treatment with the same values during 12 months of treatment. RESULTS: 45 patients (27 men, median age 29 years) completed 1-year treatment. Median weight increased from 63.1 kg in the pre-treatment period to 63.9 kg during treatment (p=0.01). Median slope of decline in lung function increased from pre-treatment FEV1 -4.1% and FVC -3.0% to +0.8% (p<0.001) and +1.6% (p=0.01), respectively. 90% of sputum samples contained mucoid P. aeruginosa before treatment, decreasing to 81% during treatment (p=0.003). Median CRP decreased from 6.2 mmol/l to 5.8 mmol/l (ns). CONCLUSION: Long-term, low-dose AZ treatment in adult CF patients with chronic P. aeruginosa infection is safe and reduces the decline in lung function, increases weight, and reduces the percentage of mucoid strains of P. aeruginosa in sputum samples. PMID- 15752680 TI - Prospective evaluation of emerging bacteria in cystic fibrosis. AB - BACKGROUND: Bacteria contribute considerably to the progression of lung disease in cystic fibrosis. In this prospective, multi-centre study, we aimed to evaluate the occurrence of emerging bacteria and the physicians' assessments of the clinical importance of these findings. METHODS: Twelve CF centres (total number of patients: 1419) reported the detection of any Stenotrophomonas maltophilia, Burkholderia cepacia complex, MRSA, Alcaligenes xylosoxidans, Klebsiella species and Mycobacteria during an observation period of 6 months. RESULTS: 213 specimens with emerging bacteria were reported from 145 different patients. The proportion of patients with emerging bacteria differed between centres (3-38%, mean: 12.6%) and increased with age. The predominant bacterium was S. maltophilia (n=106 positive specimens), followed by Klebsiellae (n=36), B. cepacia complex (n=31), A. xylosoxidans (n=16), Mycobacteria (n=11), MRSA (n=11), and others (n=2). In many instances the same microorganisms had already been reported earlier, indicating intermittent or chronic colonisation. The clinical status was reported to be stable in 70% of patients, and antibiotic treatment was anticipated for 46% of positive specimens. Comparison of clinical data to age matched controls did not reveal any significant differences with regard to pulmonary and nutritional status prior to detection of emerging bacteria. CONCLUSION: These data suggest a high variability between centres regarding the prevalence of emerging bacteria. Most patients maintained a stable clinical condition during the 6-month study period despite being colonised with emerging bacteria. PMID- 15752682 TI - Asians with cystic fibrosis in the UK have worse disease outcomes than clinic matched white homozygous delta F508 controls. AB - BACKGROUND: We tested the hypothesis that the Asian cystic fibrosis (CF) phenotype is comparable to the commonest genetic form of CF found in 50% of the white UK CF population using the UK CF Database, a national disease-specific patient registry. METHODS: 50 Asian CF patients were matched by Centre with 143 white homozygous delta F508 patients for gender, age and chronic Pseudomonas aeruginosa status (a marker of morbidity). The authors compared FEV1 and FVC% predicted, mean height, weight and BMI Z scores. RESULTS: FVC% predicted, weight and BMI Z scores were significantly worse in the Asians. Asian male/female FVC% predicted (p-value, 95% CI) -15.1 (p=0.001, -24.0, -8.8)/-15.2 (p=0.014, -27.1, 3.3) compared with white controls. Asian females also had significantly worse FEV1% predicted compared with controls (-14.9, p=0.025, 95% CI: -27.8, -2.0). Asians had significantly lower raw Z scores for weight (males p=0.002, females p=0.013) and BMI (males p=0.002, females p=0.008). CONCLUSIONS: These data suggest that the Asian CF phenotype is as severe as the white controls with the homozygous delta F508 phenotype but is worse in some outcomes, especially in Asian females. Socio-cultural factors and rare CF genotypes may contribute to the severity of CF in this vulnerable group. PMID- 15752681 TI - Risk factors for acquisition of methicillin-resistant Staphylococcus aureus (MRSA) by patients with cystic fibrosis. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is an increasing problem for patients with cystic fibrosis (CF). It has been associated with clinical deterioration in some patients with CF, creates additional infection control problems, and may affect acceptance onto transplant waiting lists. Recent attempts to eradicate the organism have met with only moderate success. An understanding of those factors which increase the risk of acquisition of MRSA by CF patients will aid the development of effective preventative strategies. We conducted a retrospective case-control study comparing a variety of risk factors for 15 MRSA-positive patients and 30 age-sex-matched MRSA-negative controls who attended the Regional Paediatric or Regional Adult Cystic Fibrosis Units in Leeds. During the year prior to initial isolation, MRSA-positive CF patients spent more days in hospital (mean 19.8 days versus 5.5 days, p=0.0003), received more treatment days of oral ciprofloxacin (43.5 days versus 13.9 days, p=0.03) more treatment days of oral/intravenous cephalosporins (42.7 days versus 15.4 days, p=0.04) and were more likely to be chronically infected with Aspergillus fumigatus (40% versus 10%, p=0.04) than the age-sex-matched MRSA-negative controls. There were no significant differences in observed clinical parameters (clinical and X-ray scores) with between the two groups. Minimising the number and length of hospital admissions and judicious use of antibiotics, particularly ciprofloxacin, should be the key components of any strategies designed to reduce the risk of MRSA acquisition by patients with CF. PMID- 15752683 TI - Associations between clinical variables and quality of life in adults with cystic fibrosis. AB - BACKGROUND: The disease progression of cystic fibrosis (CF) is marked by an increase in clinical conditions and therapeutic interventions, which have the potential to affect health-related quality of life (HRQoL). This cross-sectional study explored associations between clinical variables and HRQoL. METHODS: HRQoL was measured using the Cystic Fibrosis Quality of Life (CFQoL) questionnaire, which consists of nine domains: physical, social, treatment, chest symptoms, emotional functioning, concerns for the future, relationships, body image, and career concerns. The CFQoL was completed by 223 adults with CF. Clinical and demographic data collected were: age, gender, FEV1% predicted, BMI, Burkholderia cepacia status, lung transplant status, diabetic status, level of nutritional intervention, and presence of an intravenous access device. Multiple regression using forward selection was used to construct models relating these variables to each HRQoL domain. RESULTS: Despite many of the variables being inter-related, some variables were associated with CFQoL domains even in the presence of other important clinical factors. FEV1% predicted was weakly positively associated with all nine domains. Strong evidence emerged that patients who had received a lung transplant reported a higher HRQoL in physical and social functioning, chest symptoms, and treatment issues. Females tended to report a lower quality of life for chest symptoms and career issues, but higher values for body image. Patients with an access device expressed more career concerns. There was no evidence of an association between B. cepacia and any of the nine CFQoL domains. The model for the body image domain explained a high percentage of the variance (R2=30%): negative body image was associated with lower BMI, having an access device, diabetes, and enteral feeding. CONCLUSIONS: While important associations were identified, much of the variance in HRQoL remains unexplained. Other clinical and psychosocial variables merit investigation. A longitudinal study is required to investigate how the disease trajectory and associated treatments affect an individual's quality of life. PMID- 15752684 TI - A pilot study of oral L-arginine in cystic fibrosis. AB - Exhaled nitric oxide has previously been found to be low in cystic fibrosis. The aim of this study was to determine whether exhaled nitric oxide levels would increase in response to oral L-arginine supplementation administered daily for 4 weeks. Exhaled and nasal nitric oxide was measured weekly. Plasma L-arginine levels increased in response to supplementation but this was not reflected in an increase in eNO levels. PMID- 15752685 TI - Rapidly progressive lung disease in a patient with cystic fibrosis on long-term azithromycin: possible role of mycoplasma infection. AB - Macrolides is effective therapy in patients with cystic fibrosis (CF). We describe a girl with CF given long-term azithromycin who died of rapidly progressive lung disease. She was found to have rising titers of mycoplasma serology, suggesting a possible causative role of a resistant mycoplasma infection. Mycoplasma infection should be considered in CF patients who are deteriorating, even if they are being treated with macrolides, to which these organisms are usually susceptible. PMID- 15752686 TI - Effect of the polypeptide binding on the thermodynamic stability of the substrate binding domain of the DnaK chaperone. AB - The effect of polypeptide binding on the stability of the substrate binding domain of the molecular chaperone DnaK has been studied by thermodynamic analysis. The calorimetric scan of the fragment of the substrate binding domain DnaK384-638, consisting of a beta-domain and an alpha-helical lid, showed two transitions centered at 56.2 and 76.0 degrees C. On the other hand, the thermal unfolding of the shorter fragment DnaK386-561, which lacks half of the alpha helical lid, exhibited a single transition at 57.0 degrees C. Therefore, the transition of DnaK384-638 at 56.2 degrees C is mainly attributed to the unfolding of the beta-domain. The calorimetric scan of DnaK384-638D526N showed that the unfolding of the beta-domain was composed of two transitions. The polypeptide bound DnaK384-638 exhibited a symmetrical DSC peak at 58.6 degrees C, indicating that the substrate binding shifts the beta-domain toward a single cooperative unit. A low concentration of GdnHCl (<1.0 M) induced a conformational change in the beta-domain of DnaK384-638 without changes in the secondary structure. While the thermal unfolding of the beta-domain of DnaK384-638 was composed of two transitions in the presence of GdnHCl, the beta-domain of the substrate bound DnaK384-638 exhibited a single symmetrical DSC peak in the same condition. All together, our results indicate that complex between DnaK384-638 and substrate forms a rigid conformation in the beta-domain. PMID- 15752687 TI - Characterization of a membrane-associated apoplastic lipoxygenase in Phaseolus vulgaris L. AB - An extracytoplasmic 86.7 kDa protein was isolated from intercellular washing fluids (IWF) of Phaseolus vulgaris etiolated hypocotyls. Micro sequencing of tryptic peptides of the 86.7 kDa protein revealed 100% identity with a bean lipoxygenase (LOX) protein fragment. Purified P87-LOX exhibited LOX activity characterized by an optimal pH of 6.0 and linolenic acid as an optimal substrate, and was classified as a 13-LOX with respect to its positional specificity of linoleic acid oxygenation. A protein identical to P87-LOX, as determined by MALDI TOF analysis and biochemical characterization, was purified from hypocotyl microsomes. Immunoblot analysis showed that P87-LOX is present in plasma membrane enriched fractions, from which it was solubilized using high ionic strength buffers. These observations suggest that P87-LOX is a peripheral protein associated to the apoplastic face of the plasma membrane. PMID- 15752688 TI - Changing the metal ion selectivity of rabbit muscle enolase by mutagenesis: effects of the G37A and G41A mutations. AB - During the reaction catalyzed by enolase, a mobile loop, residues 36-45, closes over the active site. In order to probe the role of this loop movement in catalysis, the glycines at positions 37 and 41 of rabbit muscle enolase (beta beta) have been mutated to alanines. The mutant forms-G37A and G41A-of enolase are both active, but have altered selectivity for divalent cations. G37A, when assayed with Mg(2+), has 12% the activity of the wild type. However, it is twice as active as wild type when assayed with Mn(2+), Zn(2+), or Co(2+). G41A has 4% the activity of the wild type with Mg(2+), is more active than wild type with Co(2+), and slightly less active than wild type with Mn(2+) and Zn(2+). The kinetic isotope effect for both mutants is greater than that of the wild type with all 4 divalent cations. These results indicate that the flexibility of this loop has subtle effects on catalytic activity. PMID- 15752690 TI - Cloning and expression of ostrich trypsinogen: an avian trypsin with a highly sensitive autolysis site. AB - One of ostrich (Struthio camelus) trypsinogen genes was cloned from pancreatic cDNA. Its amino acid sequence compared to known trypsin sequences from other species shows high identity and suggests that it is a member of the phylogenetically anionic trypsinogen I subfamily. After cytoplasmic over expression in Escherichia coli and renaturation, the activation properties of ostrich trypsinogen were studied and compared to those of human trypsinogen 1 (also called as human cationic trypsinogen). Ostrich trypsinogen undergoes bovine enterokinase activation and autoactivation much faster than human trypsinogen 1 and exhibits on a synthetic substrate a somewhat higher enzymatic activity than the latter one. The most interesting property of ostrich trypsin is its relatively fast autolysis that can be explained via a mechanism different from the common mechanism for rat and human 1 trypsins. The latter proteases have a site, Arg117-Val118, where the autolysis starts and then goes on in a zipper-like fashion. This is absent from ostrich trypsin. Instead it has a couple of cleavage sites within regions 67-98, including two unusual ones, Arg76-Glu77 and Arg83 Ser84. These appear to be hydrolysed fast in a non-consecutive manner. Such an autolysis mechanism could not be inhibited by a single-site mutation which in humans is proposed to lead to pancreatitis. PMID- 15752689 TI - Characterization and kinetic analysis of enzyme-substrate recognition by three recombinant lactococcal tripeptidases. AB - Tripeptidases from Lactococcus lactis subsp. lactis (L9PepTR), L. lactis subsp. cremoris (L6PepTR), and L. lactis subsp. hordniae (hTPepTR) were cloned, overexpressed, purified, and characterized. Although these enzymes contained three to seven naturally occurring amino acid differences, both metal-binding and catalytic sites were highly conserved. The k(cat) values of hTPepTR were approximately 1.5- to 2-fold higher than those of L9PepTR, while, for L6PepTR, they were approximately 0.8- to 1.4-times the L9PepTR values. The K(m) of tripeptidase from subsp. lactis (L9PepTR) was considerably larger when glycine was the amino acid located at both the N- and C-terminus of the peptide substrate. In addition, the K(m) values of L9PepTR increased in the following order for YGG, LGG, FGG, SGG, and alpha-aminoisobutyrylglycylglycine, while the k(cat)/K(m) decreased in the same order. These results suggest that the dipole moment and steric hindrance of the N-terminal amino acid side chain may be the most important factors controlling substrate specificity. PMID- 15752691 TI - Processing of chicken progastrin at post-Phe bonds by an aspartyl protease. AB - Prohormones mature to biologically active peptide hormones through posttranslational modifications, which include endoproteolytic cleavages. Cleavages at mono- and dibasic sites are well characterized, and several of the responsible prohormone convertases have been identified. There is, however, evidence that endoproteolytic maturation occurs also at other sites. Among these, post-Phe cleavage occurs in the maturation of chicken progastrin, where the processing to gastrin-30 has been examined in detail. In this study we have characterized an endoprotease of the aspartic acid protease family in chicken and human tissue capable of cleaving at the Phe site. Enzymatic activity was monitored by radioimmunoassays using antibodies specific for the N- and C-termini exposed after cleavage. Analysis showed that only pepstatin, a specific inhibitor of aspartic proteases, inhibited the enzyme. The pH optimum of the enzyme ranged from pH 2 to pH 5. Amino acid substitution from Phe to Ala in the substrate completely abolished enzyme activity. The endoproteolytic activity was identified in chicken antrum and pectoral muscle as well as human cardiac and prostate extracts, suggesting that the enzyme has widespread biological functions. Experiments using recombinant cathepsin D and E indicated that neither is responsible for the endoproteolytic cleavage of chicken progastrin at post-Phe bonds. PMID- 15752692 TI - Differential scanning calorimetry of the irreversible denaturation of Rapana thomasiana (marine snail, Gastropod) hemocyanin. AB - The thermal denaturation of the hemocyanin from gastropod Rapana thomasiana (RtH) at neutral pH was studied by means of differential scanning calorimetry (DSC). The denaturation was completely irreversible as judged by the absence of any endotherm on rescanning of previously scanned samples. Two transitions, with apparent transition temperatures (T(m)) at 83 and 90 degrees C, were detected by DSC using buffer 20 mM MOPS, containing 0.1 M NaCl, 5 mM CaCl(2) and 5 mM MgCl(2), pH 7.2. Both T(m) were dependent on the scanning rate, suggesting that the thermal denaturation of RtH is a kinetically controlled process. The activation energy (E(A)) of 597+/-20 kJ mol(-1) was determined for the main transition (at 83 degrees C). E(A) for the second transition was 615+/-25 kJ mol( 1). The T(m) and Delta H(cal) values for the thermal denaturation of RtH were found to be independent of the protein concentration, signifying that the dissociation of the protein into monomers does not take place before the rate determining state of the process of thermal unfolding. PMID- 15752693 TI - Characterization of recombinant human protein C inhibitor expressed in Escherichia coli. AB - The serine protease inhibitor (serpin) protein C inhibitor (PCI; also named plasminogen activator inhibitor-3) regulates serine proteases in hemostasis, fibrinolysis, and reproduction. The biochemical activity of PCI is not fully defined partly due to the lack of a convenient expression system for active rPCI. Using pET-15b plasmid, Ni(2+)-chelate and heparin-Sepharose affinity chromatography steps, we describe here the expression, purification and characterization of wild-type recombinant (wt-rPCI) and two inactive mutants, R354A (P1 residue) and T341R (P14 residue), expressed in Escherichia coli. Wild type rPCI, but not the two mutants, formed a stable bimolecular complex with thrombin, activated protein C and urokinase. In the absence of heparin, wt-rPCI thrombin, -activated protein C, and -urokinase inhibition rates were 56.7, 3.4, and 2.3 x 10(4) M(-1) min(-1), respectively, and the inhibition rates were accelerated 25-, 71-, and 265-fold in the presence of 10 mug/mL heparin for each respective inhibition reaction. The stoichiometry of inhibition (SI) for wt-rPCI thrombin was 2.0, which is comparable to plasma-derived PCI. The present report describes for the first time the expression and characterization of recombinant PCI in a bacterial expression system and demonstrates the feasibility of using this system to obtain adequate amounts of biologically active rPCI for future structure-function studies. PMID- 15752694 TI - Compact molten globule-like state of hUBF HMG Box1 at extremely low pH. AB - Using far and near-UV CD, ANS fluorescence and 2D NMR spectroscopy, an acid induced partly folded state (A state) at extremely low pH for hUBF HMG Box1 was identified and characterized. As compared to the native state (N), the A state has similar secondary structure, less compact pack with larger amounts of exposed hydrophobic surface, and narrower chemical shift dispersion in (1)H-(15)N HSQC spectrum, which implies that it is a molten globule (MG)-like species. On the other hand, substantial tertiary contacts and cooperative thermal denaturing transition indicate that the A state is closer-relative to the classic MG-to the native folded state. In addition, when the solution pH is adjusted to neutrality, the protein in the A state refolds to the native state easily. All these data suggest that the A state of hUBF HMG Box1 could represent a potential folding intermediate on protein folding pathway. PMID- 15752695 TI - Unfolding and inactivation of cutinases by AOT and guanidine hydrochloride. AB - We present a comparative analysis of the unfolding and inactivation of three cutinases in the presence of guanidine hydrochloride (GdnHCl) and bis(2 ethylhexyl) sodium sulfosuccinate (AOT). Previous investigations have focused on the cutinase from Fusarium solani pisi (FsC). In addition to FsC, the present study includes the cutinase from Humicola insolens (HiC) and a mutant variant of HiC (muHiC) with increased activity and decreased surfactant sensitivity. Equilibrium and time-resolved denaturation by AOT were studied in aqueous solution and reverse micelles, and were compared with GdnHCl denaturation. The far-UV CD and fluorescence denaturation profiles obtained in the aqueous solutions of the two denaturants coincide for all three cutinases, indicating that unfolding is a co-operative two-state process under these conditions. In reverse micelles, the cutinases unfold with mono-exponential rates, again indicating a two-state process. The free energy of denaturation in water was calculated by linear extrapolation of equilibrium data, yielding very similar values for the three cutinases with averages of -11.6 kcal mol(-1) and -2.6 kcal mol(-1) for GdnHCl and AOT, respectively. Hence, the AOT denatured state (D(AOT)) is less destabilised than the GdnHCl denatured state (D(GdnHCl)), relative to the native state in water. Far-UV CD spectroscopy revealed that D(AOT) retains some secondary structure, while D(GdnHCl) is essentially unstructured. Similarly, fluorescence data suggest that D(AOT) is more compact than D(GdnHCl). Activity measurements reveal that both D(AOT) and D(GdnHCl) are practically inactive (catalytic activity <1% of that of the native enzyme). The fluorescence spectrum of D(AOT) in reverse micelles did not differ significantly from that observed in aqueous AOT. NMR studies of D(AOT) in reverse micelles indicated that the structure is characteristic of a molten globule, consistent with the CD and fluorescence data. PMID- 15752696 TI - The binding of 1,8 ANS congeners to I-FABP and comparison of some hypotheses about ANS' spectral sensitivity to environment. AB - The ANS congeners 1-anilinonaphthalene and 1-amino,8-sulfonato naphthalene were investigated as analogs of 1,8 anilinonaphthalene sulfonate. Like 1,8 ANS, they also bind to I-FABP, and the fully-bound spectra reveal interesting similarities and differences with respect to ANS binding. The nature of these differences suggests that certain hypotheses in the literature about ANS photophysics ought to be revised. The conceptual decomposition of energetic effects in the thermodynamics of ANS binding proposed earlier [W. Kirk, E. Kurian, F. Prendergast, Characterization of the sources of protein-ligand affinity: 1 sulfonato-8-(1')anilinonaphthalene binding to intestinal fatty acid binding protein. Biophys. J. (1996) 70 69-83.] is extended further in this report. PMID- 15752697 TI - Log P effect of organic solvents on a thermophilic alcohol dehydrogenase. AB - An alcohol dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernix was activated by water-miscible organic solvents. This activation was influenced by the kind and the concentration of the added organic solvents. The k(cat) was increased by a factor of over ten when the mole fraction of acetonitrile was 0.1. This effect was large when organic solvents with large log P values were added. In fact, the k(cat) showed a strong positive correlation with the log P value of the mixed solvent at a constant mole fraction of water, while it was not affected by the kind of organic solvents added. Both the activation enthalpy and the entropy decreased with an increase in log P. The contribution of the activation enthalpy to the free energy of activation was larger than that of the activation entropy, and the free energy of activation decreased with an increase in log P. PMID- 15752698 TI - A comparison of solution conformation and hydrodynamic properties of equine, porcine and rabbit serum albumin using viscometric measurements. AB - This paper presents the results of viscosity determinations on aqueous solutions of equine, porcine and rabbit serum albumin over a wide range of concentrations and at temperatures ranging from 5 degrees C to (42-45) degrees C. The results are compared with human and bovine serum albumin previously studied. Viscosity temperature dependence is discussed on the basis of the modified Arrhenius formula. The effective specific volume, the activation energy and entropy of viscous flow for all investigated albumins are compared. Viscosity-concentration dependence, in turn, is discussed on the basis of Mooney equation. Based on the assumption that theoretical and experimental values of Simha factor--at high temperature limit--are equal to each other, the hydrodynamic volume of the studied albumins has been calculated. The numerical values of a self-crowding factor were also obtained. At low concentration limit, the numerical values of the intrinsic viscosity and of Huggins coefficient were compared. PMID- 15752700 TI - Comparative study of the stability of poplar plastocyanin isoforms. AB - The stability of the two isoforms of poplar plastocyanin (PCa and PCb) was studied with differential scanning calorimetry (DSC) technique. It was shown that the thermal unfolding of both isoforms is an irreversible process with two endothermic and one exothermic peaks. The melting temperature of PCb was found to be 1.3+/-0.2 K degrees higher than of PCa, which indicates that PCb is more stable. The enthalpy of unfolding was estimated from the heat capacity curves and was found to be significantly higher for PCb at salt concentration I=0.1 M. In addition, PCb unfolding enthalpy and melting temperature are much more sensitive to the changes in the salt concentration as found in the experiments done at different ionic strength. The experiments were complemented with numerical calculations. The salt effect on the stability was modeled using the X-ray structure of PCa and a homology modeled structure of PCb. It was found, in agreement with the experimental data, that the stability of PCb changes by 4.7 kJ more than PCa, as the salt concentration increases from zero to 0.1 M. Thus, the differences in only 12 amino acid positions between "a" and "b" isoforms result in a measurable difference in the folding enthalpy and a significant difference in the salt dependence. The optimization of the electrostatic energies of PCa and PCb were studied and it was shown that PCb is better electrostatically optimized. PMID- 15752699 TI - The first archaeal agmatinase from anaerobic hyperthermophilic archaeon Pyrococcus horikoshii: cloning, expression, and characterization. AB - Agmatinase is one of the key enzymes in the biosynthesis of polyamines such as putrescine and sperimidine from arginine in microorganisms. The gene (PH0083) encoding the putative agmatinase of hyperthermophilic archaeon Pyrococcus horikoshii was identified based on the genome database. The gene was cloned and expressed, and the product was mainly obtained as inactive inclusion body in Escherichia coli. The inclusion body was dissolved in 6 M guanidine-HCl and successively refolded to active enzyme by the dilution of the denaturant. The enzyme exclusively catalyzed the hydrolysis of agmatine, but not arginine. This indicates that PH0083 codes agmatinase. The enzyme required divalent cations such as Co(2+), Ca(2+) and Mn(2+) for the activity. The highest activity was observed under fairly alkaline conditions, like pH 11. The purified recombinant enzyme consisted of four identical subunits with a molecular mass of 110-145 kDa. The enzyme was extremely thermostable: the full activity was retained on heating at 80 degrees C for 10 min, and a half of the activity was retained by incubation at 90 degrees C for 10 min. From a typical Michaelis-Menten type kinetics, an apparent K(m) value for agmatine was determined to be 0.53 mM. Phylogenic analysis revealed that the agmatinase from P. horikoshii does not belong to any clusters of enzymes found in bacteria and eukarya. This is the first description of the presence of archaeal agmatinase and its characteristics. PMID- 15752701 TI - A survey for phosphoglucose isomerase with lysyl aminopeptidase activity in Vibrionaceae and non-Vibrio pathogens. AB - Phosphoglucose isomerase (PGI) with a novel lysyl aminopeptidase (LysAP) activity was recently purified and characterized from Vibrio vulnificus. We showed that it cleaves the amino-terminal lysyl residue from des-Arg(10)-kallidin to produce des Arg(9)-bradykinin, suggesting that it plays a role in virulence. A survey was conducted to determine the presence of this potential virulence-enhancing enzyme among twenty-three halotolerant human and fish pathogens from eleven species within the Vibrionaceae family, including V. vulnificus, V. parahaemolyticus, V. cholerae, Aeromonas hydrophila, and Plesiomonas shigelloides. In addition, fourteen species of non-Vibrionaceae pathogens were screened for LysAP activity. Cell lysates were partially purified by anion exchange chromatography and fractions were screened for LysAP and isomerase activities. PGI-LysAP activity was detected in chromatographic fractions from all the Vibrio species tested, but was not detected in any of the non-Vibrionaceae pathogens. Levels of isomerase and LysAP activity correlated (R(2)=0.92) for nine strains of V. vulnificus. Since the Vibrionaceae represent an important family of human and fish pathogens, our identification of PGI-LysAP activity in a broad array of vibrios may lead to the development of improved analytical methods for their identification as well as interventions to reduce the high morbidity and mortality associated with some Vibrionaceae infections in clinical, veterinary, and aquaculture settings. PMID- 15752702 TI - The target of RNAIII-activating protein (TRAP) from Staphylococcus aureus: purification, crystallization and preliminary X-ray analysis. AB - The target of the RNAIII-activating protein (TRAP) is a 21 kDa protein in which phosphorylation is activated by the RNAIII-activating protein (RAP), which causes an increase in RNAII and RNAIII synthesis and the production of the virulence factors. In an attempt to examine the structural role of TRAP in the signal transduction pathway, TRAP from Staphylococcus aureus was overexpressed, purified and crystallized using PEG 8000 and 5% Jeffamine M600 (pH 7.0), as precipitants by hanging-drop vapour diffusion methods at 287 K. The crystals belong to the orthorhombic space group, P2(1)2(1)2(1), with unit cell parameters of a=39.68, b=50.41, c=85.45 A. There is one monomer of TRAP per crystallographic asymmetric unit with a crystal volume per protein mass (V(M)) of 2.06 A(3) Da(-1) and a solvent content of 40.3%. A complete data set diffracting to 1.9 A resolution was collected from a single crystal at 100 K using a synchrotron-radiation source. PMID- 15752703 TI - 1,2,5-Thiadiazolidin-3-one 1,1-dioxide-based heterocyclic sulfides are potent inhibitors of human tryptase. AB - We describe herein the design, synthesis, and in vitro biochemical evaluation of a series of potent, time-dependent inhibitors of the mast cell-derived serine protease tryptase. The inhibitors were readily obtained by attaching various heterocyclic thiols, as well as a basic primary specificity residue P(1), to the 1,2,5-thiadiazolidin-3-one 1,1-dioxide scaffold. The inhibitors were found to be devoid of any inhibitory activity toward a neutral (elastase) or cysteine (papain) protease, however they were also fairly efficient inhibitors of bovine trypsin. The differential inhibition observed with trypsin suggests that enzyme selectivity can be optimized by exploiting differences in the S' subsites of the two enzymes. The results described herein demonstrate the versatility of the heterocyclic scaffold in fashioning mechanism-based inhibitors of neutral, basic, and acidic (chymo)trypsin-like serine proteases. PMID- 15752705 TI - Identification and characterization of the ATP-binding site in human pancreatic glucokinase. AB - The central role of human pancreatic glucokinase in insulin secretion and, consequently, in maintenance of blood glucose levels has prompted investigation into identification of ATP-binding site residues and examination of ATP- and glucose-binding interactions. Because glucokinase has been resistant to crystallization, computer generated homology models were developed based on the X ray crystal structure of the COOH-terminal domain of human brain hexokinase 1 bound to glucose and ADP or glucose and glucose-6-phosphate. Human pancreatic glucokinase mutants were designed based upon these models and on ATPase domain sequence conservation to identify and characterize potential glucose and ATP binding sites. Specifically, mutants Asp78Ala, Thr82Ala, Lys90Ala, Lys102Ala, Gly227Ala, Thr228Ala, Ser336Leu, Ser411Ala, and Ser411Leu were constructed, expressed, purified, and kinetically characterized under steady-state conditions. Compared to their respective wild type controls, several mutants demonstrated dramatic changes in V(max), cooperativity of glucose binding and S(0.5) for ATP and glucose. Results suggest a role for Asp78, Thr82, Gly227, Thr228, and Ser336 in ATP binding and indicate these residues are essential for glucose phosphorylation by human pancreatic glucokinase. PMID- 15752704 TI - Cloning and characterization of histamine dehydrogenase from Nocardioides simplex. AB - Histamine dehydrogenase (NSHADH) can be isolated from cultures of Nocardioides simplex grown with histamine as the sole nitrogen source. A previous report suggested that NSHADH might contain the quinone cofactor tryptophan tryptophyl quinone (TTQ). Here, the hdh gene encoding NSHADH is cloned from the genomic DNA of N. simplex, and the isolated enzyme is subjected to a full spectroscopic characterization. Protein sequence alignment shows NSHADH to be related to trimethylamine dehydrogenase (TMADH: EC 1.5.99.7), where the latter contains a bacterial ferredoxin-type [4Fe-4S] cluster and 6-S-cysteinyl FMN cofactor. NSHADH has no sequence similarity to any TTQ containing amine dehydrogenases. NSHADH contains 3.6+/-0.3 mol Fe and 3.7+/-0.2 mol acid labile S per subunit. A comparison of the UV/vis spectra of NSHADH and TMADH shows significant similarity. The EPR spectrum of histamine reduced NSHADH also supports the presence of the flavin and [4Fe-4S] cofactors. Importantly, we show that NSHADH has a narrow substrate specificity, oxidizing only histamine (K(m)=31+/-11 microM, k(cat)/K(m)=2.1 (+/-0.4)x10(5)M(-1)s(-1)), agmatine (K(m)=37+/-6 microM, k(cat)/K(m)=6.0 (+/-0.6)x10(4)M(-1)s(-1)), and putrescine (K(m)=1280+/-240 microM, k(cat)/K(m)=1500+/-200 M(-1)s(-1)). A kinetic characterization of the oxidative deamination of histamine by NSHADH is presented that includes the pH dependence of k(cat)/K(m) (histamine) and the measurement of a substrate deuterium isotope effect, (D)(k(cat)/K(m) (histamine))=7.0+/-1.8 at pH 8.5. k(cat) is also pH dependent and has a reduced substrate deuterium isotope of (D)(k(cat))=1.3+/-0.2. PMID- 15752706 TI - Inhibition of COX activity by NSAIDs or ascorbate increases cAMP levels and enhances differentiation in 1alpha,25-dihydroxyvitamin D3-induced HL-60 cells. AB - Arachidonic acid metabolism is modulated during differentiation induced by 1alpha,25(OH)(2)D(3) in HL-60 cells. Antioxidants that affect arachidonic acid metabolism enhance this differentiation program. Ascorbate also enhances differentiation in 1alpha,25(OH)(2)D(3)-induced cells depending on the induction of cAMP. The aim of this work was to study if this cAMP rise depends on modulation of arachidonic acid metabolism by ascorbate. Cyclooxygenase inhibitors, indomethacin and aspirin, increased cAMP levels and also enhanced 1alpha,25(OH)(2)D(3)-induced differentiation in HL-60 cells. Ascorbate did not affect the release of arachidonic acid-derived metabolites but decreased the levels of TXB(2) and PGE(2), suggesting the inhibition of cyclooxygenase. On the other hand, free arachidonic acid increased both cAMP levels and differentiation in the absence or presence of 1alpha,25(OH)(2)D(3). Neither cyclooxygenase inhibitors nor ascorbate modified AA effect. Then, inhibition of cyclooxygenase activity by ascorbate could accumulate free arachidonic acid or other metabolites that increase cAMP levels and enhance differentiation in 1alpha,25(OH)(2)D(3) induced HL-60 cells. PMID- 15752707 TI - The influence of substrate on the spectral properties of oxyferrous wild-type and T252A cytochrome P450-CAM. AB - To probe whether the nature of the substrate can directly influence the spectral properties of oxyferrous cytochrome P450-CAM, the complex has been investigated in the absence and in the presence of the natural substrate (1R)-camphor (camphor) and of several camphor analogs. The oxyferrous complex of T252A P450 CAM, a mutant lacking the hydroxyl group that forms a hydrogen bond to the heme iron-coordinated dioxygen, has also been studied to gauge the influence of this hydrogen bond. UV-visible absorption and magnetic circular dichroism (MCD) spectra of these oxyferrous adducts prepared and stabilized at -40 degrees C in 60% (v/v) ethylene glycol are generally similar, exhibiting absorption bands at approximately 355, approximately 420, approximately 554, and approximately 585 nm (shoulder) and a characteristic MCD trough at approximately 585 nm. The MCD spectrum of camphor-bound oxyferrous P450-CAM is similar to that of the substrate free oxyferrous enzyme, but the spectrum of the oxyferrous enzyme differs detectably in the presence of substrate analogs. The spectra of the oxyferrous T252A mutant and wild-type enzyme are overall similar except for Soret band position blue shifts by 2-6 nm for the mutant. 5-Methylenylcamphor (epoxidation substrate) appears to have an anomalous binding mode for the mutant compared with that for the wild-type enzyme. The present results indicate that the structures of the camphor analogs can sensitively influence the physical (spectroscopic) properties of the P450 dioxygen complex and could also affect its reactivity. The ability of substrate to modulate the reactivity of P450 intermediates could be a relevant factor in explaining the remarkable diversity of reactions catalyzed by the enzyme. PMID- 15752708 TI - Expression patterns of mouse and human CYP orthologs (families 1-4) during development and in different adult tissues. AB - The present study compared the relative expression pattern of 10 orthologous CYP forms from families 1-4 in cDNA panels of human and mouse fetal and adult tissues. Except for CYP1A2, all of these CYPs exhibited specific patterns of expression during mouse ontogeny, suggesting possible involvement in development. Cyp1a1 and Cyp2r1 were the only two of the orthologs to be expressed only in the E7 mouse; Cyp2s1 was expressed in all stages, including E7, while Cyp2e1 appeared only at E17. Highest expression of the individual CYPs in the different late term human fetal tissues was: thymus, CYP1B1 and CYP2U1; liver, CYP2E1; brain, CYP2R1, CYP1A1 and CYP4X1; and lung, CYP4B1 and CYP2W1. In general, the level of individual human CYP transcripts was lower in the fetal than the corresponding adult tissues. The pattern of expression in adult mouse and human tissues was fairly similar for CYP1A1, CYP1A2, CYP1B1, CYP2S1, and CYP2U1 orthologs. PMID- 15752710 TI - Activation of deoxycytidine kinase by deoxyadenosine: implications in deoxyadenosine-mediated cytotoxicity. AB - The inborn deficiency of adenosine deaminase is characterised by accumulation of excess amounts of cytotoxic deoxyadenine nucleotides in lymphocytes. Formation of dATP requires phosphorylation of deoxyadenosine by deoxycytidine kinase (dCK), the main nucleoside salvage enzyme in lymphoid cells. Activation of dCK by a number of genotoxic agents including 2-chlorodeoxyadenosine, a deamination resistant deoxyadenosine analogue, was found previously. Here, we show that deoxyadenosine itself is also a potent activator of dCK if its deamination was prevented by the adenosine deaminase inhibitor deoxycoformycin. In contrast, deoxycytidine was found to prevent stimulation of dCK by various drugs. The activated form of dCK was more resistant to tryptic digestion, indicating that dCK undergoes a substrate-independent conformational change upon activation. Elevated dCK activities were accompanied by decreased pyrimidine nucleotide levels whereas cytotoxic dATP pools were selectively enhanced. dCK activity was found to be downregulated by growth factor and MAP kinase signalling, providing a potential tool to slow the rate of dATP accumulation in adenosine deaminase deficiency. PMID- 15752709 TI - Histone acetyltransferase p300 promotes the activation of human WT1 promoter and intronic enhancer. AB - The Wilms' tumor gene-1 (WT1) encodes a zinc finger protein involved in gene regulation during kidney, gonad, and heart development. In addition to its promoter, a 258 bp intronic enhancer is required for tissue-specific expression of WT1 gene. p300 is a histone acetyltransferase (HAT) and exerts essential functions in gene regulation. Here, we show that p300 increased the expression of endogenous WT1 mRNA and promoted the activation of the WT1 promoter and intronic enhancer. The results also revealed that the adenovirus E1A repressed the p300 function, while the p300-binding defective E1A delta 2-36 did not, and p300 HAT activity was important for its function since p300 mutant with the HAT domain deleted partially abrogated its ability to activate the WT1 promoter and intronic enhancer. Furthermore, p300 and c-Myb synergistically activated the expression of WT1 gene. This study revealed that p300 and its HAT activity were involved in regulation of WT1 transcription. PMID- 15752711 TI - Osmolyte-induced folding enhances tryptic enzyme activity. AB - Osmolytes form a class of naturally occurring small compounds known to protect proteins in their native folded and functional states. Among the osmolytes, trimethylamine-N-oxide (TMAO) has received special interest lately because it has shown an extraordinary capability to support folding of denatured to native-like species, which show significant functional activity. Most enzymes and/or proteins are commonly stored in glycerol to maintain their activity/function. In the present study, we tested whether TMAO can be a better solute than glycerol for two commonly used proteases, trypsin and chymotrypsin. Our enzyme kinetic data suggest that the enzyme activity of trypsin is significantly enhanced in TMAO compared to glycerol, whereas chymotrypsin activity is not significantly changed in either case. These results are in accordance with the osmolyte effects on the folding of these enzymes, as judged by data from fluorescence emission spectroscopy. These results suggest that TMAO may be a better solute than glycerol to maintain optimal tryptic enzyme activity. PMID- 15752712 TI - Functional importance of Ca2+-deficient N-terminal lobe of molluscan troponin C in troponin regulation. AB - Ca(2+)-binding sites I and II in the N-terminal lobe of molluscan troponin C (TnC) have lost the ability to bind Ca(2+) due to substitutions of the amino acid residues responsible for Ca(2+) liganding. To evaluate the functional importance of the Ca(2+)-deficient N-terminal lobe in the Ca(2+)-regulatory function of molluscan troponin, we constructed chimeric TnCs comprising the N-terminal lobes from rabbit fast muscle and squid mantle muscle TnCs and the C-terminal lobe from akazara scallop TnC, TnC(RA), and TnC(SA), respectively. We characterized their biochemical properties as compared with those of akazara scallop wild-type TnC (TnC(AA)). According to equilibrium dialysis using (45)Ca(2+), TnC(RA), and TnC(SA) bound stoichiometrically 3 mol Ca(2+)/mol and 1 mol Ca(2+)/mol, respectively, as expected from their primary structures. All the chimeric TnCs exhibited difference-UV-absorption spectra at around 280-290 nm upon Ca(2+) binding and formed stable complexes with akazara scallop troponin I, even in the presence of 6M urea, if Ca(2+) was present. However, when the troponin complexes were constructed from chimeric TnCs and akazara scallop troponin T and troponin I, they showed different Ca(2+)-regulation abilities from each other depending on the TnC species. Thus, the troponin containing TnC(SA) conferred as high a Ca(2+) sensitivity to Mg-ATPase activity of rabbit actomyosin-akazara scallop tropomyosin as did the troponin containing TnC(AA), whereas the troponin containing TnC(RA) conferred virtually no Ca(2+) sensitivity. Our findings indicate that the N-terminal lobe of molluscan TnC plays important roles in molluscan troponin regulation, despite its inability to bind Ca(2+). PMID- 15752713 TI - Porin and cytochrome oxidase containing contact sites involved in the oxidation of cytosolic NADH. AB - Cytochrome c (cyto-c) added to isolated mitochondria promotes the oxidation of extra-mitochondrial NADH and the reduction of molecular oxygen associated to the generation of an electrochemical membrane potential available for ATP synthesis. The electron transport pathway activated by exogenous cyto-c molecules is completely distinct from the one catalyzed by the respiratory chain. Dextran sulfate (500 kDa), known to interact with porin (the voltage-dependent anion channel), other than to inhibit the release of ATP synthesized inside the mitochondria, greatly decreases the activity of exogenous NADH/cyto-c system of intact mitochondria but has no effect on the reconstituted system made of mitoplasts and external membrane preparations. The results obtained are consistent with the existence of specific contact sites containing cytochrome oxidase and porin, as components of the inner and the outer membrane respectively, involved in the oxidation of cytosolic NADH. The proposal is put forward that the bi-trans-membrane electron transport chain activated by cytosolic cyto-c becomes, in physio-pathological conditions: (i) functional in removing the excess of cytosolic NADH; (ii) essential for cell survival in the presence of an impairment of the first three respiratory complexes; and (iii) an additional source of energy at the beginning of apoptosis. PMID- 15752714 TI - Characterization of monochloramine toxicity on PC12 cells and protective effect of tocopherol via antioxidative function. AB - Monochloramine (NH(2)Cl) is a physiological oxidant produced by activated neutrophils. In the present work, we studied the underlying mechanism of cytotoxic effects of NH(2)Cl on an undifferentiated rat pheochromocytoma PC12 cell line and the protective effects of antioxidants. The cells treated with 100 microM NH(2)Cl exhibited signs of apoptotic cell death such as phosphatidylserine exposure and caspase activation. To understand the mechanism of NH(2)Cl cytotoxicity, we examined the effect of various kinds of antioxidants including alpha-tocopherol (alpha-Toc) and beta-tocopherol (beta-Toc). These antioxidants exerted a protective effect against NH(2)Cl-induced cell death, and alpha-Toc exhibited the most potent inhibitory effect among the antioxidants used. A loss of cellular glutathione was observed in the cells treated with 100 microM NH(2)Cl. The formation of reactive oxygen species (ROS) was also measured using the fluorescent probe dichlorofluorescin diacetate. The fluorescence intensity increased prior to cell death and an antioxidant, such as alpha-Toc, suppressed the increase in ROS. Interestingly, beta-Toc also exerted similar inhibitory effects on cytotoxicity and caspase activation. These results suggest that free radical mediated process is involved in NH(2)Cl-induced PC12 cell death and that tocopherols inhibit this cell death via antioxidative function. PMID- 15752715 TI - A retinoic acid binding cytochrome P450: CYP120A1 from Synechocystis sp. PCC 6803. AB - At least 35 cytochrome P450 (P450, CYP) or cytochrome P450-like genes have been identified in 10 cyanobacterial genomes yet none have been functionally characterized. CYP110 and CYP120 represent the two largest cyanobacterial P450 families with 16 and four members, respectively, identified to date. The Synechocystis sp. PCC 6803 CYP120A1 protein sequence shares high degrees of conservation with CYP120A2 from Trichodesmium erythraeum IMS101 and CYP120B1 and CYP120C1 from Nostoc punctiforme PCC 73102. In this communication, we report the cloning, expression, purification, and characterization of CYP120A1 from Synechocystis. Homology modeling predictions of the three-dimensional structure of CYP120A1 coupled with in silico screening for potential substrates and experimental spectroscopic analyses have identified retinoic acid as a compound binding with high affinity to this P450's catalytic site. These characterizations of Synechocystis CYP120A1 lay the initial foundations for understanding the basic role of cytochrome P450s in cyanobacteria and related organisms. PMID- 15752716 TI - Role of sulfhydryl groups in band 3 in the inhibition of phosphate transport across erythrocyte membrane in visceral leishmaniasis. AB - Membrane destabilization in erythrocytes plays an important role in the premature hemolysis and development of anemia during visceral leishmaniasis (VL). Marked degradation of the anion channel protein band 3 is likely to allow modulation of anion flux across the red cell membrane in infected animals. The present study describes the effect of structural modification of band 3 on phosphate transport in VL using (31)P NMR. The result showed progressive decrease in the rate and extent of phosphate transport during the post-infection period. Interdependence between the intracellular ionic levels seems to be a determining factor in the regulation of anion transport across the erythrocyte membrane in control and infected conditions. Infection-induced alteration in band 3 made the active sites of transport more susceptible to binding with amino reactive agents. Inhibition of transport by oxidation of band 3 and subsequent reversal by reduction using dithiothreitol suggests the contribution of sulfhydryl group in the regulation of anion exchange across the membrane. Quantitation of sulfhydryl groups in the anion channel protein showed the inhibition to be closely related to the decrease of sulfhydryl groups in the infected hamsters. Downregulation of phosphate transport during leishmanial infection may be ascribed to the sulfhydryl modification of band 3 resulting in the impaired functioning of this protein under the diseased condition. PMID- 15752717 TI - Gap junction channels reconstituted in two closely apposed lipid bilayers. AB - Intercellular communication mediated by gap junction channels plays an important role in many cellular processes. In contrast to other channels, gap junction channels span two plasma membranes resulting in an intracellular location for both ends of the junctional pore and the regulatory sites for channel gating. This configuration presents unique challenges for detailed experimental studies of junctional channel physiology and ligand-activation in situ. Availability of an appropriate model system would significantly facilitate future studies of gap junction channel function and structure. Here we show that the double-membrane channel can be reconstituted in pairs of closely apposed lipid bilayers, as experienced in cells. We have trapped the calcium-sensitive dye, arsenazo III (AIII), partially calcium-saturated (AIII-Ca), in one population of connexin32 reconstituted-liposomes, and EGTA in a second one. In such mixtures, the interaction of EGTA with AIII-Ca was measured by a large color shift from blue to red (decreased absorbance at 652 nm). The exchange of these compounds through gap junctions was proportional to these decrements. Results indicate that these connexon-mediated interliposomal channels are functional and are inhibited by the addition of alpha-glycyrrhetinic acid and by flufenamic acid, two gap junction communication inhibitors. Future use of this model system has the potential to improve our understanding of the permeability and modulation of junctional channels in its native intercellular assembly. PMID- 15752718 TI - Differential changes in phospholipase D and phosphatidate phosphohydrolase activities in ischemia-reperfusion of rat heart. AB - Phospholipase D (PLD2) produces phosphatidic acid (PA), which is converted to 1,2 diacylglycerol (DAG) by phosphatidate phosphohydrolase (PAP2). Since PA and DAG regulate Ca(2+) movements, we examined PLD2 and PAP2 in the sarcolemma (SL) and sarcoplasmic reticular (SR) membranes from hearts subjected to ischemia and reperfusion (I-R). Although SL and SR PLD2 activities were unaltered after 30 min ischemia, 5 min reperfusion resulted in a 36% increase in SL PLD2 activity, whereas 30 min reperfusion resulted in a 30% decrease in SL PLD2 activity, as compared to the control value. SR PLD2 activity was decreased (39%) after 5 min reperfusion, but returned to control levels after 30 min reperfusion. Ischemia for 60 min resulted in depressed SL and SR PLD2 activities, characterized with reduced V(max) and increased K(m) values, which were not reversed during reperfusion. Although the SL PAP2 activity was decreased (31%) during ischemia and at 30 min reperfusion (28%), the SR PAP2 activity was unchanged after 30 min ischemia, but was decreased after 5 min reperfusion (25%) and almost completely recovered after 30 min reperfusion. A 60 min period of ischemia followed by reperfusion caused an irreversible depression of SL and SR PAP2 activities. Our results indicate that I-R induced cardiac dysfunction is associated with subcellular changes in PLD2 and PAP2 activities. PMID- 15752719 TI - Reversible activation of secretory phospholipase A2 by sulfhydryl reagents. AB - Secretory phospholipase A(2)s (sPLA(2)s) have been implicated in physiological and pathological events, but the regulatory mechanism(s) of their activities in cells remains to be solved. Previously, we reported that phenylarsine oxide (PAO), a sulfhydryl reagent, stimulated arachidonic acid (AA) release in rat pheochromocytoma PC12 cells. In this study, we examined the effects of thimerosal, another sulfhydryl reagent, to clarify the sulfhydryl modification and activation of sPLA(2) molecules in cells. Like PAO, thimerosal-stimulated AA release in an irreversible manner and the responses were not additive. Dithiol compounds such as dithiothreitol inhibited AA release from both the thimerosal- and the PAO-treated cells, and monothiol compounds (l-Cys and glutathione) decreased the thimerosal response. Both sulfhydryl reagents stimulated AA release from the HEK293T cells expressing human sPLA(2)X, and stimulated the sPLA(2) activities of bee venom sPLA(2) and the soluble fraction of sPLA(2)X-expressing cells. Our results suggest that the sPLA(2)s in cells are inactive and modification of disulfide bonds in the molecules can be a trigger of sPLA(2) activation in cells. Sulfhydryl reagents are useful tools for studying the regulatory mechanism(s) of sPLA(2) activity in cells. PMID- 15752720 TI - Equilibrium titrations of acid-induced unfolding-refolding and salt-induced molten globule of cytochrome c by FT-IR spectroscopy. AB - Despite extensive investigations on the acid-unfolded and acid/salt-induced molten globule(-like) states of cytochrome c using variety of techniques, structural features of the acid-unfolded state in terms of residual secondary structures and the structural transition between the acid-unfolded and acid/salt refolded states have not been fully characterized beyond the circular dichroism (CD) spectroscopy. It is unusual that secondary structure(s) of the unfolded state leading to the molten globule state, an important protein folding intermediate, as determined by CD was not fully corroborated by independent experimental method(s). In this study, we carried out an equilibrium titration of acid-induced unfolding and subsequent acid- and salt-induced refolding of cytochrome c using Fourier transform infrared spectroscopy. The spectral profiles of the equilibrium titration reveal new structural details about the acid unfolded state and the structural transition associated with the acid/salt refolded molten globule(-like) states of cytochrome c. PMID- 15752721 TI - Functional characterizations of novWUS involved in novobiocin biosynthesis from Streptomyces spheroides. AB - NovW, novU, and novS gene products represent dTDP-4-keto-6-deoxy-D-glucose 3,5 epimarase, C-methyltransferase and dTDP-glucose-4-ketoreductase involved in noviose biosynthetic pathway, respectively. We have expressed three genes to elucidate the functions of NovW, NovU, and NovS in Escherichia coli. NovW and NovU catalyze the formation of dTDP-4-keto-6-deoxy-5-C-methyl-L-lyxo-hexose from dTDP-4-keto-6-deoxy-D-glucose. NovS reduces the product formed from the reaction of NovW with dTDP-4-keto-6-deoxy-D-glucose in the presence of NADH to result in dTDP-l-rhamnose. Furthermore, a pathway for the biosynthesis of noviose is proposed. PMID- 15752722 TI - On the difference in stability between horse and sperm whale myoglobins. AB - The work in the literature on apomyoglobin is almost equally divided between horse and sperm whale myoglobins. The two proteins share high homology, show similar folding behavior, and it is often assumed that all folding phenomena found with one protein will also be found with the other. We report data at equilibrium showing that horse myoglobin was 2.1 kcal/mol less stable than sperm whale myoglobin at pH 5.0, and aggregated at high concentrations as measured by gel filtration and analytical ultracentrifugation experiments. The higher stability of sperm whale myoglobin was identified for both apo and holo forms, and was independent of pH from 5 to 8 and of the presence of sodium chloride. We also show that the substitution of sperm whale myoglobin residues Ala15 and Ala74 to Gly, the residues found at positions 15 and 74 in horse myoglobin, decreased the stability by 1.0 kcal/mol, indicating that helix propensity is an important component of the explanation for the difference in stability between the two proteins. PMID- 15752723 TI - Examination of MgATP binding in a tryptophan-shift mutant of phosphofructokinase from Bacillus stearothermophilus. AB - A tryptophan-shift variant of Bacillus stearothermophilus phosphofructokinase (BsPFK), W179F/F76W, was constructed to evaluate the binding and allosteric characteristics associated with MgATP. W179F/F76W BsPFK has a specific activity of 77+/-1 U/mg at pH 7 and 25 degrees C, which is a 35% decrease compared to the wild-type enzyme. The K(m) for MgATP increases from 43+/-3 microM for wild-type BsPFK to 160+/-7 microM in the variant. Binding and allosteric interaction between Fru-6-P and PEP for the variant are similar to those of the wild-type enzyme. W179F/F76W BsPFK has distinct fluorescence properties relative to wild type or other tryptophan-shifted mutants of BsPFK. The binding of MgATP produces an 80% decrease in the fluorescence intensity while MgADP causes a 70% decrease. Capitalizing on these fluorescence changes, dissociation constants of 30+/-1 microM and 0.53+/-0.02 mM were measured for MgATP and MgADP, respectively. In addition, PEP was shown to enhance MgATP binding by 2.6-fold. PMID- 15752724 TI - Cathepsin B promotes both motility and invasiveness of oral carcinoma cells. AB - We previously demonstrated that overexpression of cathepsin B (CB) protease in oral squamous cell carcinomas correlated positively with advanced tumor stage and poor histologic malignancy grade. Here we examined whether CB contributes to the invasiveness of oral carcinoma cells. For RNA-mediated inhibition, two ribozymes that target CB mRNA were designed and stably expressed in the oral squamous cell carcinoma cell line 1386Tu. Both ribozymes diminished expression of CB mRNA, protein, and activity, without affecting cathepsin D or beta-actin, as determined by quantitative real-time PCR, Western blots, and protease activity assays. Matrigel invasion assays showed that the invasiveness of the cells was significantly reduced by the expressed ribozymes and, surprisingly, the motilities of the ribozyme-transfected cells were also diminished. Our results document a direct role for CB in promoting oral cancer spread and invasion, and open the possibility of controlling oral carcinoma malignancy and metastasis by targeting CB with RNA inhibitor strategies. PMID- 15752725 TI - Measurement and characterization of C-3 epimerization activity toward vitamin D3. AB - Recently, epimerization of the hydroxyl group at C-3 has been identified as a unique metabolic pathway of vitamin D compounds. We measured C-3 epimerization activity in subcellular fractions prepared from cultured cells and investigated the basic properties of the enzyme responsible for the epimerization. C-3 epimerization activity was detected using a NADPH-generating system containing glucose-6-phosphate, NADP, glucose-6-phosphate dehydrogenase, and Mg(2+). The highest level of activity was observed in a microsomal fraction prepared from rat osteoblastic UMR-106 cells but activity was also observed in microsomal fractions prepared from MG-63, Caco-2, Hep G2, and HUH-7 cells. In terms of maximum velocity (V(max)) and the Michaelis constant (K(m)), 25-hydroxyvitamin D(3) [25(OH)D(3)] exhibited the highest specificity for the epimerization at C-3 among 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], 25(OH)D(3), 24,25 dihydroxyvitamin D(3) [24,25(OH)(2)D(3)], and 22-oxacalcitriol (OCT). The epimerization activity was not inhibited by various cytochrome P450 inhibitors and antiserum against NADPH cytochrome P450 reductase. Neither CYP24, CYP27A1, CYP27B1 nor 3(alpha-->beta)hydroxysteroid epimerase (HSE) catalyzed the epimerization in vitro. Based on these results, the enzyme(s) responsible for the epimerization of vitamin D(3) at C-3 are thought to be located in microsomes and different from cytochrome P450 and HSE. PMID- 15752726 TI - Ferredoxin-dependent glutamate synthase moonlights in plant sulfolipid biosynthesis by forming a complex with SQD1. AB - UDP-sulfoquinovose synthase, SQD1, catalyzes the transfer of sulfite to UDP glucose giving rise to UDP-sulfoquinovose, which is the head group donor for the biosynthesis of the plant sulfolipid sulfoquinovosyldiacylglyerol. The native SQD1 enzyme of spinach exists as a 250 kDa heteroprotein complex with much higher affinity for the substrate sulfite than the recombinant SQD1 protein itself. The SQD1 protein co-purified with nine proteins. Likely binding partners included rubisco activase, HSP70, and ferredoxin-dependent glutamate synthase (FdGOGAT). While the first two proteins are known to interact with many other proteins, the identification of FdGOGAT was most intriguing because this 160kDa protein contains an FMN cofactor known to bind sulfite in vitro. Using different constructs expressing recombinant forms of the multidomain protein FdGOGAT, it was demonstrated that the FMN-binding domain of FdGOGAT is essential for specific binding of the protein to SQD1. A model suggests that FdGOGAT could channel sulfite to SQD1. PMID- 15752727 TI - Regulation of tyrosinase by tetrahydropteridines--what is real? A comment on the work published by Wood et al. on December 24, 2004. PMID- 15752729 TI - Functional characterization of a conditionally immortalized mouse epididymis caput epithelial cell line MEPC5 using temperature-sensitive simian virus 40 large T-antigen. AB - A conditionally immortalized epididymis caput cell line, MEPC5, was established by infecting primary cultured mouse epididymis caput cells with a temperature sensitive simian virus 40 large T-antigen. At a permissive temperature of 33 degrees C, the large T-antigen was expressed and the cells grew continuously. However, the downregulation of T-antigen at a nonpermissive temperature of 39 degrees C and the upregulation of cell density at 33 degrees C were associated with growth arrest and the increased protein expression of p21(waf1), a cell cycle inhibitor. The cells expressed epididymal caput-expressed genes such as phosphatidylethanolamine binding protein, polyoma enhancer activator 3, ME1, sulfated glycoprotein-2 (SGP-2), androgen receptor, and retinoic acid receptor alpha. Interestingly, the expression levels of ME1 and SGP-2 were significantly elevated under the cell growth-restricted conditions. The established mouse epididymis caput epithelial cell line MEPC5 retains some characteristics of differentiated epididymis epithelial cells, and should prove an excellent model for studies of gene expression and the physiological functions of epididymis caput epithelial cells. PMID- 15752728 TI - Role of phospholipase D1 in neurite outgrowth of neural stem cells. AB - Employing neural stem cells from the brain cortex of E12 rat embryos, we investigated the possible role of phospholipase D (PLD) in the synaptogenesis and neurite formation of neural cells during differentiation. Expression level of PLD1 increased during neuronal differentiation of the neural stem cells, resulting in increased PLD activity. Expression level of synapsin I, a marker of synaptogenesis, also increased as the differentiation of neural stem cells progressed. To figure out the effect of PLD on synapsin I expression, we treated the neural stem cells with phorbol myristate acetate (PMA) to stimulate PLD activity. Increased PLD activity induced by PMA treatment resulted in elevated synapsin I expression and neurite outgrowth during neuronal differentiation. To further confirm the role of PLD in neurite outgrowth, we transfected the dominant negative form of rat PLD1 cDNA (DN-rPLD1) into neural stem cells to downregulate PLD activity. Overexpression of DN-rPLD1 showed the complete inhibition of neurite outgrowth of neural stem cells under differentiation condition. While transfection of DN-rPLD1 did not affect the synapsin I expression, overexpression of rPLD1 resulted in increased synapsin I expression of the neural cells. These results suggest that PLD1 plays a critical role in neurite outgrowth during differentiation of the neural stem cells. In conclusion, this is the first evidence to show that PLD1 acts as an important regulator of neurite outgrowth in neural stem cell by promoting neuronal differentiation via increase of synapsin I expression. PMID- 15752730 TI - Comparative genomics approach toward critical determinants for the imprinting of an evolutionarily conserved gene Impact. AB - The Impact is an evolutionarily conserved gene subjected to genomic imprinting in mouse but not in human. A characteristic tandem repeat similar to those found in many other imprinted genes and an elevated expression level, both observed only for the mouse gene, are implicated in the evolution of imprinting, to which the repeat might have contributed via enhancement of the expression. To pursue the possibility further, we examined the correlation among the repeat, expression level, and imprinting of Impact in various mammals ranging from rodents, lagomorphs, carnivores, artiodactyls to primates. Intriguingly, rabbit Impact is abundantly expressed and imprinted like those of rodents, but is missing the repeat from its first intron like those of other mammals that express both alleles weakly. It thus seems that lineage-specific enhancement of gene expression rather than the tandem repeat per se played a critical role in the evolution of imprinting of Impact. PMID- 15752731 TI - Purification and partial characterization of a low temperature responsive Mn-SOD from tea (Camellia sinensis (L.) O. Kuntze). AB - The manganese containing superoxide dismutase (Mn-SOD) was purified from a tea clone, TEENALI, which showed the lowest period of winter dormancy. Protein was purified using leaves of tea by ammonium sulfate precipitation, followed by column chromatography using DEAE-cellulose, and silica-based size exclusion chromatography on HPLC system. Upto 51-fold purification and a specific activity of 56.66 U/mg of protein was achieved, which yielded a single band upon denaturing PAGE. The enzyme had a native molecular weight of about 169 kDa, whereas a monomer with molecular weight of 43 kDa was found on SDS-PAGE suggesting it to be homotetramer. The purified enzyme had pH optima of 8.0. It exhibited a wide temperature range for its activity with optima at 0 degrees C suggesting its role in low temperature tolerance. The manuscript presents purification and characterization of high molecular weight Mn-SOD from tea and discusses its implication in tolerance of low temperature stress. PMID- 15752733 TI - Novel TLR4-antagonizing peptides inhibit LPS-induced release of inflammatory mediators by monocytes. AB - Toll-like receptor 4 (TLR4) has become a new target for combating Gram-negative bacterium-induced sepsis. In this study, we screened peptides that can interact with TLR4 from a random 16-peptide library using yeast two-hybrid system and performed functional identification for the obtained peptides. We got two positive clones out of 1.28x10(7) transformants. The peptides were sequenced and synthesized. Protein sequence comparison confirmed that the two peptides had no homologous proteins. The two peptides were found to significantly inhibit LPS induced NF-kappaB activation in HEK-293 cells that were transfected with TLR4 cDNA, LPS-induced IkappaBalpha (IkappaB alpha) phosphorylation and NF-kappaB activation in monocytes, and release of IL-1, IL-6, and TNF-alpha by monocytes. We further confirmed that the No. 9 peptide could bind to TLR4 extracellular domain, but the No. 24 peptide could not, suggesting that two novel peptides were identified as the antagonists of TLR4, which significantly inhibited the effects of endotoxin in vitro. The No. 9 peptide may function through binding to TLR4 extracellular domain. Our findings suggest a promising countermeasure against Gram-negative bacterium-induced sepsis. PMID- 15752732 TI - Interleukin-4 inhibits RANKL-induced expression of NFATc1 and c-Fos: a possible mechanism for downregulation of osteoclastogenesis. AB - Interleukin-4 (IL-4), an anti-inflammatory cytokine, has been shown to inhibit osteoclast differentiation. Therefore, this cytokine is considered to be a promising therapeutic applicant for bone-resorbing diseases such as rheumatoid arthritis (RA). Recently NFATc1, a transcription factor, has been shown to play critical roles in osteoclastogenesis. The aim of this study was to clarify the role of IL-4 on the intracellular signaling of NFATc1. A RAW264.7 monocyte/macrophage cell line and murine bone marrow precursors were differentiated into osteoclasts in the presence of receptor activator of nuclear factor kappaB ligand (RANKL) and/or macrophage colony-stimulating factor. Tartrate-resistant acid phosphatase (TRAP) staining and a pit assay using dentine were used for the identification of activated osteoclasts. The protein expression of IL-4 receptor, NFATc1, and c-Fos was determined by Western blot analysis. In addition, the gene expression of NFATc1 and c-Fos was determined by reverse transcription and polymerase chain reaction. The IL-4 receptor was constitutively expressed in RAW264.7 cells. RANKL induced osteoclast generation, as determined by TRAP staining and pit assay. IL-4 inhibited RANKL-induced osteoclastogenesis at low concentrations of 10ng/ml and more. Interestingly, IL-4 potently inhibited RANKL-induced expression of NFATc1 at mRNA level. Furthermore, IL-4 inhibited c Fos expression, which is shown to be responsible for NFATc1 expression, in time- and dose-dependent manners. In addition, IL-4 inhibited the RANKL-induced expression of NFATc1 and c-Fos in murine bone marrow cells. Thus, we suggest that IL-4 may downregulate osteoclastogenesis in part through inhibition of the expression of transcription factors, NFATc1 and c-Fos. These findings provide new insight into development of new medication for osteoporosis and RA. PMID- 15752734 TI - P120-GAP associated with syndecan-2 to function as an active switch signal for Src upon transformation with oncogenic ras. AB - BALB/3T3 cells transfected with plasmids pcDNA3.1-[S-ras(Q(61)K)] of shrimp Penaeus japonicus were applied to reveal a complex of p120-GAP/syndecan-2 being highly expressed upon transformation. Of interest, most of the p120-GAP/syndecan 2 complex was localized at caveolae, a membrane microdomain enriched with caveolin-1. To confirm the molecular interaction between syndecan-2 and p120-GAP, we further purified p120-GAP protein from mouse brains by using an affinity column of HiTrap-RACK1 and expressed mouse RACK1-encoded fusion protein and mouse syndecan-2-encoded fusion protein in bacteria. We report molecular affinities exist between p120-GAP and RACK1, syndecan-2 and RACK1 as well as p120-GAP and syndecan-2. The selective affinity between p120-GAP and syndecan-2 was found to be sufficient to detach RACK1. The p120-GAP/syndecan-2 complex was demonstrated to keep Src tyrosine kinase in an activated form. On the other hand, the syndecan 2/RACK1 complex was found to have Src in an inactivated form. These data indicate that the p120-GAP/syndecan-2 complex at caveolae could provide a docking site for Src to transmit tyrosine signaling, implying that syndecan-2/p120-GAP functions as a tumor promoter upon transformation with oncogenic ras of shrimp P. japonicus. PMID- 15752735 TI - A gene cluster for the fatty acid catabolism from Pseudonocardia autotrophica BCRC12444. AB - Genes involved in fatty acid degradation (fad) were isolated from Pseudonocardia autotrophica BBRC12444. Six open reading frames and a bi-directional promoter region were identified by DNA sequence analyses and primer extension. The fad gene cluster included five ORFs, designated fadA, fadB, fadR, fadC, and fadD. Base on their amino acid sequence identity, the gene products were identified as acyl-CoA ligase (FadA), enoyl-CoA hydratase (FadB), transcriptional regulator (FadR), cytochrome P450 monooxygenase (FadC), and ferredoxin (FadD). Regulatory protein, FadR, could bind to an operator sequence located in the divergent promoter region between fadR and fadC genes, implicating the control of fatty acid degradation. The real-time quantitative PCR assays revealed that the expression of the fadA, fadB, fadR, and fadC genes was induced by long chain fatty acids and repressed by glucose. All results demonstrated that the fad gene cluster participated in the pathway of the fatty acid catabolism. This is the first bacterial fad gene cluster to be reported. PMID- 15752736 TI - A comparison of two DNA base excision repair glycosylases from Arabidopsis thaliana. AB - Plants contain the genes for both formamidopyrimidine-DNA glycosylase (FPG) and oxoguanine glycosylase (OGG). These enzymes play analogous roles in the base excision repair pathways of bacteria (FPG) and archaea, yeast, and mammals (OGG). Why have plants retained both genes? We tested one hypothesis by comparing the specificities of Arabidopsis FPG and OGG purified from Escherichia coli expression clones. Using depurinated DNA as substrate, the specific activity of Arabidopsis FPG was higher than that of Arabidopsis OGG. Using DNA oxidized by treatment with light in the presence of methylene blue, the specific activities of Arabidopsis FPG and OGG were equal. Using an oligonucleotide containing one oxoguanine (paired with C) and labeled with fluorescein, the specific activity of Arabidopsis OGG was greater than that of either FPG. The results support the hypothesis that genes for the two enzymes have been retained during evolution of plants for their specialized enzyme activities. PMID- 15752737 TI - Endostatin induces acute endothelial nitric oxide and prostacyclin release. AB - Chronic exposure to endostatin (ES) blocks endothelial cell (EC) proliferation, and migration and induces EC apoptosis thereby inhibiting angiogenesis. Nitric oxide (NO) and prostacyclin (PGI(2)), in contrast, play important roles in promoting angiogenesis. In this study, we examined the acute effects of ES on endothelial NO and PGI(2) production. Unexpectedly, a cGMP reporter cell assay showed that ES-induced acute endothelial NO release in cultured bovine aortic endothelial cells (BAECs). Enzyme immunoassay showed that ES also induced an acute increase in PGI(2) production in BAECs. These results were confirmed by ex vivo vascular ring studies that showed vascular relaxation in response to ES. Immunoblot analysis showed that ES stimulated acute phosphorylation of endothelial nitric oxide synthase (eNOS) at Ser116, Ser617, Ser635, and Ser1179, and dephosphorylation at Thr497 in BAECs, events associated with eNOS activation. Short-term exposure of EC to ES, therefore, unlike long-term exposure which is anti-angiogenic, may be pro-angiogenic. PMID- 15752738 TI - Evaluation of insulin resistance linkage to rat chromosome 4 in SHR of a Japanese colony. AB - The spontaneously hypertensive rat (SHR) is a model of human insulin resistance syndrome. Quantitative trait loci for cellular defects in glucose and fatty acid metabolism have been mapped to an overlapping region of rat chromosome (RNO) RNO4 in SHR of the National Institute of Health colony, where a deletion in the Cd36 gene has been implicated as the causative mutation of insulin resistance. The present study has examined the potential presence of RNO4 linkage to a series of metabolic phenotypes in F(2) progeny derived from SHR of a Japanese colony (SHR/Izm) without the Cd36 mutation. Our data demonstrate that 'major' insulin resistance gene(s) are unlikely to exist on RNO4 in SHR/Izm and in vitro phenotypes measured in isolated adipocytes do not cosegregate in the F(2) population studied. Thus, it seems to be difficult to explain the underlying genetic mechanisms of insulin resistance by a single major gene on RNO4. PMID- 15752739 TI - Psychological stress induces chemoresistance in breast cancer by upregulating mdr1. AB - Psychological distress reduces the efficacy of chemotherapy in breast cancer patients. The mechanism may be related to the altered neuronal or hormonal secretions during stress. Here, we reported that adrenaline, a hormone mediating the biological activities of stress, upregulates mdr1 gene expression in MCF-7 breast cancer cells via alpha(2)-adrenergic receptors in a dose-dependent manner. Mdr1 upregulation can be specifically inhibited by pretreatment with mdr1-siRNA. Consequently, adrenergic stimulation enhances the pump function of P-glycoprotein and confers resistance of MCF-7 cells to paclitaxel. In vivo, restraint stress increases mdr1 gene expression in the MCF-7 cancers that are inoculated subcutaneously into the SCID mice and provokes resistance to doxorubicin in the implanted tumors. The effect can be blocked by injection of yohimbine, an alpha(2)-adrenergic inhibitor, but not by metyrapone, a corticosterone synthesis blocker. Therefore, we conclude that breast cancers may develop resistance against chemotherapeutic drugs under psychological distress by over-expressing mdr1 via adrenergic stimulation. PMID- 15752740 TI - The role of alternative splicing and C-terminal amino acids in thromboxane receptor stabilization. AB - The thromboxane receptor has two alternatively spliced isoforms, alpha and beta, which differ only in sequences within the cytoplasmic C-terminal domain. Oxidative stress induced by H(2)O(2) in a COS-7 cell model results in stabilization of the thromboxane receptor beta isoform by translocation from the endoplasmic reticulum to the Golgi complex, which in turn results in protection of the receptor from degradation. We now report that both the alpha and beta thromboxane receptor isoforms respond identically to oxidative stress. Further, mutagenesis studies indicate that replacing the normal C-terminus with a nonsense sequence also does not alter stabilization behaviour ruling out a role for the distinct C-termini in this process. Further mutagenesis implicates a cluster of arginine residues within the C-terminal domain as involved in oxidative stress induced stabilization. These data identify a region of the thromboxane receptor that is responsible for responding to oxidative challenge and open the possibility of identification of the molecular machinery underpinning this response. PMID- 15752741 TI - A co-cultured skin model based on cell support membranes. AB - Tissue engineering of skin based on collagen:PCL biocomposites using a designed co-culture system is reported. The collagen:PCL biocomposites having collagen:PCL (w/w) ratios of 1:4, 1:8, and 1:20 have been proven to be biocompatible materials to support both adult normal human epidermal Keratinocyte (NHEK) and mouse 3T3 fibroblast growth in cell culture, respectively, by Dai, Coombes, et al. in 2004. Films of collagen:PCL biocomposites were prepared using non-crosslinking method by impregnation of lyophilized collagen mats with PCL/dichloromethane solutions followed by solvent evaporation. To mimic the dermal/epidermal structure of skin, the 1:20 collagen:PCL biocomposites were selected for a feasibility study of a designed co-culture technique that would subsequently be used for preparing fibroblast/biocomposite/keratinocyte skin models. A 55.3% increase in cell number was measured in the designed co-culture system when fibroblasts were seeded on both sides of a biocomposite film compared with cell culture on one surface of the biocomposite in the feasibility study. The co-culture of human keratinocytes and 3T3 fibroblasts on each side of the membrane was therefore studied using the same co-culture system by growing keratinocytes on the top surface of membrane for 3 days and 3T3 fibroblasts underneath the membrane for 6 days. Scanning electron microscopy (SEM) and immunohistochemistry assay revealed good cell attachment and proliferation of both human keratinocytes and 3T3 fibroblasts with these two types of cells isolated well on each side of the membrane. Using a modified co-culture technique, a co-cultured skin model presenting a confluent epidermal sheet on one side of the biocomposite film and fibroblasts populated on the other side of the film was developed successfully in co-culture system for 28 days under investigations by SEM and immunohistochemistry assay. Thus, the design of a co-culture system based on 1:20 (w/w) collagen:PCL biocomposite membranes for preparation of a bi-layered skin model with differentiated epidermal sheet was proven in principle. The approach to skin modeling reported here may find application in tissue engineering and screening of new pharmaceuticals. PMID- 15752743 TI - The tyrosine kinases Fyn and Hck favor the recruitment of tyrosine-phosphorylated APOBEC3G into vif-defective HIV-1 particles. AB - The main function of Vif is to limit the antiviral activity of APOBEC3G by counteracting its packaging into HIV-1 virions. In this work, we examine the possible functional interactions between Vif, APOBEC3G, and two Src family tyrosine kinases, Fyn and Hck, present in T lymphocytes and in monocyte macrophages, respectively. By GST pull-down, we show that the SH3 domains of Fyn and Hck, and the corresponding full-length proteins bind Vif of HIV-1. One consequence of this interaction is a reduction in their catalytic activity. Interestingly, we also observed that APOBEC3G can be phosphorylated on tyrosine in the presence of Fyn or Hck, suggesting that both kinases may regulate APOBEC3G function. Accordingly, we demonstrate that in the presence of Fyn or Hck and in the absence of Vif, the overall level of APOBEC3G incorporated into HIV-1 particles is decreased, whereas the level of encapsidation of its phosphorylated form is significantly enhanced. PMID- 15752742 TI - Meg1/Grb10 overexpression causes postnatal growth retardation and insulin resistance via negative modulation of the IGF1R and IR cascades. AB - The Meg1/Grb10 protein has been implicated as an adapter protein in the signaling pathways from insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) in vitro. To elucidate its in vivo function, four independent Meg1/Grb10 transgenic mouse lines were established, and the effects of excess Meg1/Grb10 on both postnatal growth and glucose metabolism were examined. All of the Meg1/Grb10 transgenic mice showed growth retardation after weaning (3-4 weeks), which indicates that ectopic overexpression of Meg1/Grb10 inhibits postnatal growth that is mediated by IGF1 via IGF1R. In addition, the mice became hyperinsulinemic owing to high levels of insulin resistance, which demonstrates that Meg1/Grb10 also modulates the insulin receptor cascade negatively in vivo. Type II diabetes arose frequently in the two transgenic lines, which also showed impaired glucose tolerance. In these mice, severe atrophy of the pancreatic acinus cells was associated with high-level production of Meg1/Grb10 in the pancreas. These results suggest that Meg1/Grb10 inhibits the function of both insulin and IGF1 receptors in these cells, since a similar phenotype has been reported for Ir and Igf1r double knockout mice. Taken together, these results indicate that Meg1/Grb10 interacts with both insulin and IGF1 receptors in vivo, and negatively regulates the IGF growth pathways via these receptors. PMID- 15752744 TI - Induced ICER Igamma down-regulates cyclin A expression and cell proliferation in insulin-producing beta cells. AB - We have previously found that cyclin A expression is markedly reduced in pancreatic beta-cells by cell-specific overexpression of repressor inducible cyclic AMP early repressor (ICER Igamma) in transgenic mice. Here we further examined regulatory effects of ICER Igamma on cyclin A gene expression using Min6 cells, an insulin-producing cell line. The cyclin A promoter luciferase assay showed that ICER Igamma directly repressed cyclin A gene transcription. In addition, upon ICER Igamma overexpression, cyclin A mRNA levels markedly decreased, thereby confirming an inhibitory effect of ICER Igamma on cyclin A expression. Suppression of cyclin A results in inhibition of BrdU incorporation. Under normal culture conditions endogenous cyclin A is abundant in these cells, whereas ICER is hardly detectable. However, serum starvation of Min6 cells induces ICER Igamma expression with a concomitant very low expression level of cyclin A. Cyclin A protein is not expressed unless the cells are in active DNA replication. These results indicate a potentially important anti-proliferative effect of ICER Igamma in pancreatic beta cells. Since ICER Igamma is greatly increased in diabetes as well as in FFA- or high glucose-treated islets, this effect may in part exacerbate diabetes by limiting beta-cell proliferation. PMID- 15752746 TI - The substrate specificity of SARS coronavirus 3C-like proteinase. AB - The 3C-like proteinase of severe acute respiratory syndrome coronavirus (SARS) has been proposed to be a key target for structural based drug design against SARS. We have designed and synthesized 34 peptide substrates and determined their hydrolysis activities. The conserved core sequence of the native cleavage site is optimized for high hydrolysis activity. Residues at position P4, P3, and P3' are critical for substrate recognition and binding, and increment of beta-sheet conformation tendency is also helpful. A comparative molecular field analysis (CoMFA) model was constructed. Based on the mutation data and CoMFA model, a multiply mutated octapeptide S24 was designed for higher activity. The experimentally determined hydrolysis activity of S24 is the highest in all designed substrates and is close to that predicted by CoMFA. These results offer helpful information for the research on the mechanism of substrate recognition of coronavirus 3C-like proteinase. PMID- 15752745 TI - Lipophilic ionophore complexes as superoxide dismutase mimetics. AB - A wide range of metal ion complexes exhibit superoxide dismutase like activities as detected by inhibition of nitroblue tetrazolium reduction. Mn(II) and Cu(II) complexes of EDTA, EHPG, and EGTA exhibit SOD like activities commensurate with many of the purpose-built SOD mimics. Here, we report analogous lipophilic chelators that localise metal ions (Cu(II), Mn(II), and Fe(III)) in the lipid membranes and lipoproteins to protect them from superoxide mediated oxidative damage. Spectroscopic titrations and Jobs method confirm that both 1:1 and 2:1 metal ion monensin complexes form. The cupric complexes are the most active exhibiting IC(50) values of 0.09 and 0.18 microM for 2Cu(II)-monensin and Cu(II) monensin, respectively, for superoxide destruction. In addition, the IC(50) value for Mn(II)-monensin is 0.31 microM. In conclusion, Mn(II) and Cu(II) complexes of the ionophore monensin exhibit considerable superoxide scavenging activities and represent a novel class of catalytic antioxidants for the protection of lipid structures. PMID- 15752747 TI - Impaired acidification in early endosomes of ClC-5 deficient proximal tubule. AB - ClC-5 chloride channel deficiency causes proteinuria, hypercalciuria, and nephrolithiasis (Dent's disease). Impaired endosomal acidification in proximal tubule caused by reduced chloride conductance is a proposed mechanism; however, functional analysis of ClC-5 in oocytes predicts low ClC-5 chloride conductance in endosomes because of their acid interior pH and positive potential. Here, endosomal pH and chloride concentration were measured in proximal tubule cell cultures from wildtype vs. ClC-5 deficient mice using fluorescent sensors coupled to transferrin (early/recycling endosomes) or alpha(2)-macroglobulin (late endosomes). Initial pH in transferrin-labeled endosomes was approximately 7.2, decreasing at 15 min to 6.0 vs. 6.5 in wildtype vs. ClC-5 deficient cells, respectively; corresponding endosomal chloride concentration increased from approximately 16 mM to 47 vs. 36 mM. In contrast, acidification and chloride accumulation were not impaired in late endosomes or Golgi. Our results provide direct evidence for ClC-5 involvement in acidification of early endosomes in proximal tubule by a chloride shunt mechanism. PMID- 15752748 TI - Genes involved in nonpermissive temperature-induced cell differentiation in Sertoli TTE3 cells bearing temperature-sensitive simian virus 40 large T-antigen. AB - Sertoli TTE3 cells, derived from transgenic mice bearing temperature-sensitive simian virus 40 large T (tsSV40LT)-antigen, proliferated continuously at a permissive temperature (33 degrees C) whereas inactivation of the large T-antigen by a nonpermissive temperature (39 degrees C) led to differentiation as judged by elevation of transferrin. To clarify the detailed mechanisms of differentiation, we investigated the time course of changes in gene expression using cDNA microarrays. Of the 865 genes analyzed, 14 genes showed increased levels of expression. Real-time quantitative PCR revealed that the mRNA levels of p21(waf1), milk fat globule membrane protein E8, heat-responsive protein 12, and selenoprotein P were markedly elevated. Moreover, the differentiated condition induced by the nonpermissive temperature significantly increased mRNA levels of these four genes in several cell lines from the transgenic mice bearing the oncogene. The present results regarding changes in gene expression will provide a basis for a further understanding of molecular mechanisms of differentiation in both Sertoli cells and cell lines transformed by tsSV40LT-antigen. PMID- 15752749 TI - Cholesterol 7alpha-hydroxylase is phosphorylated at multiple amino acids. AB - The activity of cholesterol 7alpha-hydroxylase (gpCYP7A1), the rate limiting enzyme in bile acid synthesis, has been postulated to be regulated by phosphorylation/dephosphorylation. This study has found that several kinase activators rapidly reduce the amount of bile acid produced by the human hepatoma cell line, HepG2, and that gpCYP7A1 from HepG2 cell extracts eluted in the phosphoprotein fraction of FeIII columns. After incubating the HepG2 cells with radioactive orthophosphate, the band identified as gpCYP7Al on immunoblots was strongly labeled. Recombinant gpCYP7A was expressed as 6xHIS fusion polypeptides and subjected to kinase assays. The locations of phosphorylation were mapped further by screening synthetic peptides against AMP-activated protein kinase (AMPK), c-Jun N-terminal kinase, protein kinase A, and a panel of nine protein kinase C isoforms. AMPK, also known as 3-hydroxy-3-methylglutaryl coenzyme A reductase kinase, phosphorylated cholesterol 7alpha-hydroxylase, suggesting a potential mechanism of coordination of cholesterol synthesis and degradation. PMID- 15752750 TI - The Klebsiella pneumoniae nitrogenase Fe protein gene (nifH) functionally substitutes for the chlL gene in Chlamydomonas reinhardtii. AB - The entire coding region of chlL, an essential chloroplast gene required for chlorophyll biosynthesis in the dark in Chlamydomonas reinhardtii, was precisely replaced by either the Klebsiella pneumoniae nifH (encoding the structural component of nitrogenase Fe protein) or the Escherichia coli uidA reporter gene encoding beta-glucuronidase. Homoplasmic nifH or uidA transformants were identified by Southern blots after selection on minimal medium plates for several generations. All the uidA transformants had the "yellow-in-the-dark" phenotype characteristic of chlL mutants, whereas homoplasmic nifH transformants exhibited a partial "green-in-the-dark" phenotype. NifH protein was detected in the nifH transformants but not in the wild-type strain by Western blotting. Fluorescence emission measurements also showed the existence of chlorophyll in the dark-grown nifH transformants, but not in the dark-grown uidA transformants. The nifH transplastomic form of C. reinhardtii that lacks the chlL gene can still produce chlorophyll in the dark, suggesting that the nifH product can at least partially substitute for the function of the putative "chlorophyll iron protein" encoded by chlL. Thus, introducing nitrogen fixation gene directly into a chloroplast genome is likely to be feasible and providing a possible way of engineering chloroplasts with functional nitrogenase. Notably, to introduce foreign genes without also introducing selective marker genes, a novel two-step chloroplast transformation strategy has been developed. PMID- 15752751 TI - PDGF-D contributes to neointimal hyperplasia in rat model of vessel injury. AB - In this study, we determined the role of PDGF-D, a new member of the PDGF family, in a rat model of balloon injured artery made with a 2F catheter in Sprague Dawley male rats. PDGF-D expression was studied in the injured and control segments of abdominal aorta. The function of PDGF-D was evaluated in rat vascular smooth muscle cells stably transfected with PDGF-D gene. We found that in normal abdominal aorta, PDGF-D was highly expressed in adventia, moderate in endothelia, and unidentified in media. Stable transfection of PDGF-D gene into vascular smooth muscle cells increased the cell migration by 2.2-fold, and the proliferation by 2.3-fold, respectively, and MMP-2 production and activity as well. These results support the fact that PDGF-D is involved in the formation of neointimal hyperplasia induced by balloon catheter injury and may serve as a target in preventing vascular restenosis after coronary angioplasty. PMID- 15752752 TI - Functional characterization of the HNF4alpha isoform (HNF4alpha8) expressed in pancreatic beta-cells. AB - Mutations in the hepatocyte nuclear factor (HNF) 4alpha gene cause a form of maturity-onset diabetes of the young (MODY1), which is a monogenic form of type 2 diabetes characterized by impaired insulin secretion by pancreatic beta-cells. HNF4alpha is a transcription factor expressed in the liver, kidney, intestine, and pancreatic islet. Multiple splice variants of the HNF4alpha gene have been identified and an isoform of HNF4alpha8, an N-terminal splice variant, is expressed in pancreatic beta-cells. However, expression levels of HNF4alpha protein in pancreatic beta-cells and the transcriptional activity of HNF4alpha8 are not yet understood. In the present study, we investigated the expression of HNF4alpha in beta-cells and examined its functional properties. Western blotting and immunohistochemical analysis revealed that the expression of HNF4alpha protein in pancreatic islets and INS-1 cells was much lower than in the liver. A reporter gene assay showed that the transactivation potential of HNF4alpha8 was significantly weaker than that of HNF4alpha2, which is a major isoform in the liver, suggesting that the total level of HNF4alpha activity is very weak in pancreatic beta-cells. We also showed that the N-terminal A/B region of HNF4alpha8 possessed no activation function and C-terminal F region negatively regulated the transcriptional activity of HNF4alpha8. The information presented here would be helpful for the better understanding of MODY1/HNF4alpha diabetes. PMID- 15752753 TI - Hepatoprotective and antioxidant effects of Morus bombycis Koidzumi on CCl4 induced liver damage. AB - The antioxidant activity and liver protective effect of Morus bombycis Koidzumi were investigated. Aqueous extracts of M. bombycis Koidzumi had higher superoxide radical scavenging activity than other types of extracts. The aqueous extract at a dose of 100 mg/kg showed significant hepatoprotective activity when compared with that of a standard agent. The biochemical results were confirmed by histological observations indicating that M. bombycis Koidzumi extract together with CCl(4) treatment decreased ballooning degeneration. The water extract recovered the CCl(4)-induced liver injury and showed antioxidant effects in assays of FeCl(2)-ascorbic acid-induced lipid peroxidation in rats. Based on these results, we suggest that the hepatoprotective effect of the M. bombycis Koidzumi extract is related to its antioxidative activity. PMID- 15752754 TI - Identification and characterization of murine DNAM-1 (CD226) and its poliovirus receptor family ligands. AB - The leukocyte adhesion molecule DNAM-1 (CD226) is a member of the immunoglobulin superfamily and constitutively expressed on the majority of CD4+ and CD8+ T lymphocytes, natural killer (NK) cells, monocytes/macrophages, and a subset of B lymphocytes. The poliovirus receptor (PVR; CD155) and its family member nectin 2 (CD112) have recently been identified as the ligands for DNAM-1. Interaction of DNAM-1 with the ligands induces NK cell- and CD8+ T cell-mediated cytotoxicity and cytokine secretion. However, in vivo function of the receptor-ligand interaction has remained unclear. Here, we identified murine DNAM-1 and PVR homologues that physically and functionally bind each other. We demonstrated that ligand binding of murine DNAM-1 induced a costimulatory signal in antigen specific CD8+ T cells. These results should provide a useful animal model to explore a role of DNAM-1 in immune responses in vivo. PMID- 15752755 TI - Characterization of mouse myotilin and its promoter. AB - Myotilin is a sarcomeric protein mutated in two forms of muscle disease, limb girdle muscular dystrophy type 1A and myofibrillar myopathy. Myotilin is expressed late during human myofibrillogenesis and localizes to Z-discs in mature sarcomere. It interacts with alpha-actinin, actin, and filamin C, and has strong F-actin-bundling activity. These features suggest an important role for myotilin in sarcomere organization. In our effort towards the construction of a genetic model for myotilin-related muscle disorders, we have cloned mouse myotilin, including its promoter region, and studied the expression in various tissues. Mouse myotilin is 90% identical with the human orthologue. Northern blot analysis revealed strong mRNA transcripts in skeletal and cardiac muscle, and weak expression in liver and lung tissue. Western blot and RT-PCR analysis showed the presence of one major product in mouse tissues. Analysis of the 5'-flanking region revealed a number of putative regulatory elements that drive expression in differentiating myoblasts. Finally, endogenous myotilin is induced at later stages of Z-disc assembly in C(2)C(12) cells indicating conservation between mouse and human promoter region. PMID- 15752756 TI - Identification of novel CBP interacting proteins in embryonic orofacial tissue. AB - cAMP response element-binding protein (CREB)-binding protein (CBP) plays an important role as a general co-integrator of multiple signaling pathways and interacts with a large number of transcription factors and co-factors, through its numerous protein-binding domains. To identify nuclear factors associated with CBP in developing orofacial tissue, a yeast two-hybrid screen of a cDNA library derived from orofacial tissue from gestational day 11 to 13 mouse embryos was conducted. Using the carboxy terminus (amino acid residues 1676-2441) of CBP as bait, several novel proteins that bind CBP were identified, including an Msx interacting-zinc finger protein, CDC42 interaction protein 4/thyroid hormone receptor interactor 10, SH3-domain GRB2-like 1, CCR4-NOT transcription complex subunit 3, adaptor protein complex AP-1 beta1 subunit, eukaryotic translation initiation factor 2B subunit 1 (alpha), and cyclin G-associated kinase. Results of the yeast two-hybrid screen were confirmed by glutathione S-transferase pull down assays. The identification of these proteins as novel CBP-binding partners allows exploration of new mechanisms by which CBP regulates and integrates diverse cell signaling pathways. PMID- 15752757 TI - Construction of a novel expression system on a human artificial chromosome. AB - Efficient regulation of transgene would greatly facilitate the analysis of gene function in biological systems for basic research and clinical applications. The tetracycline-regulatable system (TRS) has proven to be a promising tool for such purposes. Despite their widespread application, a number of challenges are still associated with the use of TRS, including clonal variability in the regulation and copy number. We have recently constructed a novel human artificial chromosome (HAC) called 21DeltaqHAC. By housing a TRS-based DNA-PKcs expression cassette in this HAC, we were able to circumvent the problems associated with conventional TRS-based vectors. We achieved tight control of DNA-PKcs expression and rescued the radiosensitive phenotype of DNA-PKcs-deficient CHO cells. The combined use of HAC and the TRS serves as a model for controllable and fixed copy number expression vectors. Our study also demonstrates the suitability of the HAC to accommodate multi-subunit constructs such as that of the TRS. PMID- 15752758 TI - The effects of thiophosphate substitutions on native siRNA gene silencing. AB - RNA mediated interference has emerged as a powerful tool in controlling gene expression in mammalian cells. We investigated the gene silencing properties of six thiophosphate substituted siRNAs (all based on a commercial luciferase medium silencer) compared to that of unmodified siRNA. We also examined the cytotoxicity and dose-response using several thiophosphate modified siRNAs with unmodified siRNA. Our results show that two thiophosphate siRNA sequences convert from medium to high silencers with the addition of four randomly placed thiophosphates. Both thiophosphate siRNAs have a statistically significant difference in luciferase gene silencing (5% and 6% activity) relative to the unmodified native medium silencer referred to as siRNA-2 (18% activity) and four other thiophosphate siRNAs that maintain their medium silencing capability. This indicates that specific thiophosphate substitutions may alter native siRNA function. Further, this shows that thiophosphate siRNAs with the same nucleotide sequence but with different sulfur modification positions have different silencing effects. Both the native siRNA and the thio siRNAs showed a concentration dependent relationship, i.e., with concentration increase, the luciferase gene silencing effect also increased. Confirming cytotoxicity experiments showed no significant changes when HeLa cells were treated with 10nM thiophosphate siRNAs over the course of several days. These results suggest that specific placement of thiophosphates could play an important role in the development of siRNAs as therapeutics by engineering in properties such as strength of binding, nuclease sensitivity, and ultimately efficacy. PMID- 15752759 TI - Expression of TARSH gene in MEFs senescence and its potential implication in human lung cancer. AB - To reveal the molecular mechanism of cellular senescence, we have surveyed the genes that are specifically upregulated via MEFs senescence by suppression subtractive hybridization method. We show here that mTARSH was induced particularly in the relative early phase of MEFs cellular senescence. Further structural analysis of mTARSH disclosed five splicing variants shared a common reading frame whose diversity was derived from the SH3-binding motif cluster in the middle of the gene. We also show that mTARSH mRNA predominantly expressed in lung and that conspicuous expression of TARSH was drastically declined in all several lung cancer cell lines we tested. Thus, TARSH presumably represents a trigger gene for evoking cellular senescence, which has also been suggested to be involved in the prevention of tumorigenesis. PMID- 15752760 TI - Co-culture of human CD34+ cells with mesenchymal stem cells increases the survival of CD34+ cells against the 5-aza-deoxycytidine- or trichostatin A induced cell death. AB - It has been suggested that epigenetic regulation plays an important role in maintaining the stemness and lineage differentiation of hematopoietic stem cells (HSCs), 5-aza-deoxycytidine (aza-D) and Trichostatin A (TSA) being candidate additives for HSC ex vivo expansion. Although they have potent activity to maintain the stemness, they can also cause serious cell death. This study examined the effects of mesenchymal stem cells (MSCs) on the maintenance of CD34+ cells driven by aza-D and TSA in culture with the combined cytokines of thrombopoietin, flt-3 ligand, stem cell factor, interleukin-3, and interleukin-6. In cultures without MSCs, although aza-D and TSA retained the CD34 frequency 4 to 8 times more than in the cytokines alone, a large portion of cells underwent apoptotic cell death. Consequently, CD34+ cell expansion could not be achieved in any condition without MSCs. In cultures with MSCs, the total cell number was higher in aza-D or TSA than in any conditions in the cultures without MSCs. The CD34 frequency was also similar to the level in the cultures in aza-D or TSA without the MSCs. These results suggest that a co-culture of CD34+ cells with the MSCs might not simply deliver the proliferation signals but also stemness and survival signals, and overlap the action of epigenetic regulators. PMID- 15752761 TI - PARG1, a protein-tyrosine phosphatase-associated RhoGAP, as a putative Rap2 effector. AB - Rap2 belongs to the Ras family of small GTP-binding proteins, but its specific signaling role is unclear. By yeast two-hybrid screening, we have found that the Caenorhabditis elegans ortholog of Rap2 interacts with a protein containing a Rho GTPase-activating protein (RhoGAP) domain, ZK669.1a, whose human ortholog PARG1 exhibits RhoGAP activity in vitro. ZK669.1a and PARG1 share a homology region with previously unknown function, designated the ZK669.1a and PARG1 homology (ZPH) region. Here we show that the ZPH region of PARG1 mediates interaction with Rap2. PARG1 interacted with Rap2 in a GTP-dependent manner but not with Ras or Rap1. We also show that PARG1 and its mutant lacking the ZPH region induce typical cytoskeletal changes for Rho inactivation in fibroblasts. Rap2 suppressed this in vivo action of PARG1 but not that of the mutant PARG1. These results suggest that PARG1 is a putative specific effector of Rap2 to regulate Rho. PMID- 15752762 TI - Protection against malaria induced by chirally modified Plasmodium falciparum's MSP-1 42 pseudopeptides. AB - The C-terminal portion of the Plasmodium falciparum blood stage MSP-1 antigen plays a key role in invasion of human erythrocytes. The MSP-1(1282-1301) non polymorphic 1585 peptide, from the processed MSP-1(42) fragment, is poorly immunogenic and highly alpha-helical [Angew. Chem. Int. Ed. 40 (2001) 4654]. Assessing the alpha-carbon asymmetry and its implication in the host immune response is proposed in this work to overcome the 1585 peptide's immunological properties. Accordingly, the effect of incorporating single D-amino acids and psi [CH(2)-NH] isoster bonds into the 1585 peptide was examined both at the immunogenic and 3D-structure levels. Therefore, specific binding to RBCs is promoted by site-directed chiral modifications on the native peptide as well as by simultaneously combining specific D-substitutions with psi-[CH(2)-NH] isoster bonds transforming this molecule into a high specific HLAbeta1*1101 allele binder. D-analog pseudopeptide immunized animals induced antibodies selectively recognizing a recombinant as well as native MSP-1(42) and MSP-1(33) fragments. Protection and low parasitemia levels were induced in Aotus monkeys immunized with the EVLYL(dK)PLAGVYRSLKKQLE analog. Peptide alpha-carbon chiral transformation is therefore an important target for structural modulation and, consequently, represents a novel approach towards designing multi-component subunit-based malarial vaccines. PMID- 15752763 TI - Simvastatin triggers mitochondria-induced Ca2+ signaling alteration in skeletal muscle. AB - Statin drugs represent the major improvement in the treatment of hypercholesterolemia that constitutes the main origin of atherosclerosis, leading to coronary heart disease. Besides tremendous beneficial effects of statins, various forms of muscular toxicity (myalgia, cramp, exercise intolerance, and fatigability) occur frequently. We hypothesized that the iatrogenic effects of statins could result from alterations in Ca(2+) homeostasis. Acute applications of simvastatin on human skeletal muscle fibers triggered a Ca(2+) wave of intra cellular Ca(2+) that mostly originates from sarcoplasmic reticulum (SR) Ca(2+) release. In addition, simvastatin increased mitochondrial NADH content and induced mitochondrial membrane depolarization (EC(50)=1.96 microM) suggesting an altered mitochondrial function. Consequently on simvastatin application, a weak mitochondrial Ca(2+) efflux (EC(50)=7.8 microM) through permeability transient pore and Na(+)/Ca(2+) exchanger was triggered, preceding the large SR-Ca(2+) release. Increased SR Ca(2+) content after acute application of statin is also suggested by the increased Ca(2+) spark amplitude and by the effect of cyclopiazonic acid. We thus conclude that simvastatin induced alterations in mitochondrial function which lead to an increase in cytoplasmic Ca(2+), SR-Ca(2+) overload, and Ca(2+) waves. Taken together, these statin-induced muscle dysregulations may contribute to myotoxicity. PMID- 15752765 TI - 4,4'-Dianilino-1,1'-binaphthyl-5,5'-sulfonate, a novel molecule having chaperone like activity. AB - 4,4'-Dianilino-1,1'-binaphthyl-5,5'-sulfonate (bis-ANS) and 1-anilinonaphthalene 8-sulfonate (ANS) are hydrophobic probes that are widely used in protein folding studies, using their capacity to bind to hydrophobic regions of partially unfolded proteins and in turn leading to an increase in fluorescence. Here we reveal a novel chaperone-like activity for bis-ANS, which acted as a highly effective inhibitor for the thermal- or chemical-induced aggregation of alcohol dehydrogenase, insulin or the whole cell extract of Escherichia coli, with ANS showing a much weaker effect. The studies to elucidate the mechanism underlying this activity show that bis-ANS is able to form stable soluble aggregates with the denaturing proteins and dramatically increase its fluorescence intensity upon incubation with aggregation-prone proteins. Moreover, we found that bis-ANS is able to prevent the heat inactivation of citrate synthase. These observations suggest that bis-ANS is able to block the exposed hydrophobic surfaces to suppress protein aggregation, acting in a way similar to what small heat shock proteins (one sub-class of molecular chaperones) do. The data presented here, together with the report that bis-ANS was able to suppress the amyloid formation of the prion peptide [J. Biol. Chem. 279 (2004) 5346], suggest that this molecule may be used as a potential protein stabilizer in addition to its current application as a hydrophobic probe. PMID- 15752764 TI - Crystal structure of recombinant human growth and differentiation factor 5: evidence for interaction of the type I and type II receptor-binding sites. AB - The crystal structure of human growth differentiation factor 5 (GDF5) was solved at 2.4A resolution. The structure is very similar to the structure of bone morphogenetic factor 7 (BMP7) and consists of two banana-shaped monomers, linked via a disulfide bridge. The crystal packing of GDF5 is the same as the crystal packing of BMP7. This is highly unusual since only 25-30% of the crystal contacts involve identical residues. Analysis of the crystal packing revealed that residues of the type I receptor epitope are binding to residues of the type II receptor-binding epitope. The fact that for both BMP family members the type I and type II receptor-binding sites interact suggests that the complementary sites on the receptors may interact as well, suggesting a way how preformed receptor heterodimers may form, similar to the preformed receptors observed for the erythropoietin receptor and the BMP2 receptors. PMID- 15752766 TI - Transcriptional regulation of the mouse interleukin-2 receptor beta chain gene by Ets and Egr-1. AB - To clarify the mechanisms and factors involved in the regulation of mouse IL 2Rbeta gene expression, we isolated the 5'-flanking region of IL-2Rbeta gene and investigated the promoter activity. Here we elucidated the positive regulatory regions, the most potent of which are located between -50 to -30bp and -164 to 135bp. These regions contain a potentially functional Ets and Egr-1-binding sites whose mutations abrogate promoter activity. Data from electrophoretic mobility shift assay indicate that Ets and Egr-1, but not Sp1, bind to the positive regulatory regions, -50 to -30bp and -164 to -135bp, respectively. Furthermore, recruitment of Ets and Egr-1 at endogenous IL-2Rbeta promoter segments in an IL-2 dependent F7 cells was verified by the chromatin immunoprecipitation assay. This study for the first time delineates the molecular mechanisms underlying regulation of mouse IL-2Rbeta gene transcription by Ets family proteins, partially with Egr-1, and thereby further elucidates the molecular basis of lymphocyte activation and differentiation. PMID- 15752767 TI - Circulating adhesion molecules in apoE-deficient mouse strains with different atherosclerosis susceptibility. AB - Recruitment of inflammatory cells in the arterial wall by vascular adhesion molecules plays a key role in development of atherosclerosis. Apolipoprotein E deficient (apoE(-/-)) mice have spontaneous hyperlipidemia and develop all phases of atherosclerotic lesions. We sought to examine plasma levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) and sP-selectin in two apoE(-/-) strains C57BL/6 (B6) and BALB/c with early or advanced lesions. Mice were fed chow or a Western diet containing 42% fat, 0.15% cholesterol, and 19.5% casein. On either diet, BALB/c.apoE(-/-) mice developed much smaller atherosclerotic lesions and displayed significantly lower levels of sVCAM-1 and sP-selectin than B6.apoE(-/-) mice. The Western diet significantly elevated sVCAM-1 levels in both strains and sP-selectin levels in B6.apoE(-/-) mice. BALB/c.apoE(-/-) mice exhibited 2-fold higher HDL cholesterol levels on the chow diet and 15-fold higher HDL levels on the Western diet than B6.apoE(-/-) mice, although the two strains had comparable levels of total cholesterol and triglyceride. Thus, increased atherosclerosis is accompanied by increases in circulating VCAM-1 and P selectin levels in the two apoE(-/-) mouse strains, and the high HDL level may protect against atherosclerosis by inhibiting the expression of adhesion molecules in BALB/c.apoE(-/-) mice. PMID- 15752768 TI - Characterization of two non-testis-specific CABYR variants that bind to GSK3beta with a proline-rich extensin-like domain. AB - To explore more possible roles for GSK3beta function, the yeast two-hybrid screening using GSK3beta as a bait protein was performed. In this study, we demonstrated that two variants of CABYR (281 and 379) interacted with GSK3beta in the yeast two-hybrid and GST pull down assay. Molecular characterization showed that CABYR variants formed a dimer with a proline-rich extensin-like domain, which slightly overlapped with GSK3beta-binding site. In kinase assay, we also showed that CABYR variants act as an ideal substrate for GSK3beta within the extensin-like domain and phosphorylation sites on CABYR were mapped. Interestingly, Northern blot showed that CABYR transcripts were expressed more distinctly in the fetal brain than in the adult brain, suggesting that this protein may play a role during brain development. Moreover, differential expression of CABYR variants may exhibit aberrant expression in brain tumors and cancer cell lines. PMID- 15752769 TI - Evidence that insulin secretion influences SNAP-25 through proteasomal activation. AB - Regulation of SNARE proteins by glucose in pancreatic islets is complex and insufficiently clarified. We aimed to study effects of glucose per se separate from enhancing effects on exocytosis. A 24h culture of rat islets at elevated glucose (27 mmol/L) increased t-SNARES (SNAP-25, syntaxin) (Western blotting). Co culture with diazoxide, which inhibits glucose-induced insulin secretion, reversed these effects. Effects on SNAP-25 were similar in human and rat islets. Effects of diazoxide were mimicked by blocking secretion with somatostatin (rat islets). Blocking secretion by cooling abolished both glucose and diazoxide effects on SNAP-25. Total SNAP-25 mRNA as well as isoforms alpha and beta were increased by 24-h elevated glucose. Diazoxide failed to reverse the glucose effects on mRNA. However, effects of diazoxide on SNAP-25 protein were nullified by proteasome inhibitors (ALLN, MG-132, and epoxomicin) but not by lysosomal inhibition (NH(4)Cl). Exocytosis per se modifies SNAREs by a process linked to proteasomal activation. PMID- 15752770 TI - Regulation of adiponectin receptor R1 and R2 gene expression in adipocytes of C57BL/6 mice. AB - Adiponectin has gained significant attention as a mediator of insulin sensitivity. Recently, two receptors of this adipocyte-secreted hormone, adiponectin receptor 1 (AdipoR1) and 2 (AdipoR2), have been cloned. To improve our understanding of the regulation of these receptors in adipocytes, AdipoR1 and AdipoR2 mRNA was measured by quantitative real-time reverse transcription polymerase chain reaction in brown adipocytes and adipocytes from epigonadal and subcutaneous adipose tissue of C57BL/6 mice as a function of feeding and circadian clocks. AdipoR1 gene expression was higher in brown adipose tissue and epigonadal adipose tissue, but lower in subcutaneous fat in fasted as compared to random-fed mice. In parallel, AdipoR2 mRNA levels were also higher in epigonadal adipose tissue of fasted as compared to fed mice, however, there was no regulation of AdipoR2 mRNA in brown and subcutaneous fat depending on the feeding state. Furthermore, AdipoR2 gene expression was significantly higher in epigonadal as compared to subcutaneous fat. Interestingly, a parallel circadian gene expression pattern for both AdipoR1 and AdipoR2 with lower expression between 20:00 and 06:00 h in brown, epigonadal, and subcutaneous adipose tissue was also found. In conclusion, our results suggest a fat depot specific regulation of AdipoR1 and AdipoR2 gene expression in brown and white fat by fasting. In addition, we have identified a coordinated circadian pattern of AdipoR1 and AdipoR2 gene expression in these tissues. PMID- 15752771 TI - Evaluation of in vitro inhibitory potential of small interfering RNAs directed against various regions of foot-and-mouth disease virus genome. AB - India is endemic for foot-and-mouth disease and it continues to be a major threat to the livestock industry despite vaccination programmes. In the present study, the ability of specific small interfering (si)RNAs directed against different genomic regions of foot-and-mouth disease virus (FMDV) to inhibit virus replication in BHK-21 cells was examined. For preliminary evaluation of possible siRNA-mediated FMDV inhibition, a cocktail of several unique populations of 12 30bp siRNAs were successfully produced corresponding to three target regions located at structural (VP3-VP1), non-structural (2A-2C), and non-structural untranslated (3D-3'UTR) region of serotype Asia1. Once the populations of siRNAs generated were found to reduce the virus titre significantly, two highly conserved 21bp siRNA duplexes were designed by analysing all FMDV sequence entries available in public-domain databases. In virus titration assay, more than 99% inhibition of virus yield for all the four serotypes (type Asia1, O, A, and C) could be demonstrated in cells transfected with each of the FMDV-specific siRNAs at 24h post-infection, compared to control cells transfected with scrambled siRNA. This was well supported by reduction in OD values in FMDV specific sandwich ELISA. Although 100-fold reduction in virus titre with siRNA1 is substantial considering the transfection efficiency and fixed level of input siRNA, siRNA2 emerged to be a better choice as target where more than 300-fold reduction was observed and its inhibitory effect extended up to 48 h post infection against all the serotypes. Interestingly, in the present study type A virus (IND 17/77) had a single mismatch at position 2 in the siRNA2 target region but it did not abrogate the inhibitory effect. PMID- 15752772 TI - Intracellular interferon triggers Jak/Stat signaling cascade and induces p53 dependent antiviral protection. AB - Intracellular interferons (IFNs) exert biological functions similar to those of extracellular IFNs, but the signal transduction pathway triggered by the intracellular ligands has not been fully revealed. We investigated the signaling cascade by sequence-specific knockdown of signaling molecules by means of the RNA interference. Truncated IFN-beta gene was constructed so that the N-terminal secretory signal sequence was deleted (SD.IFN-beta). Cells transfected with this construct showed phosphorylation and activation of the STAT1 without any detectable secretion of the cytokine. The MHC class I expression was significantly augmented, while the augmentation was suppressed by short interfering RNA duplexes specific for JAK1, TYK2, and IFN-alpha/beta receptor (IFNAR) 1 and 2c chains. The SD.IFN-beta also induced p53 and phosphorylation of p53 at Ser(15). Specific silencing of p53 abrogated the antiviral effect of SD.IFN-beta, suggesting that the tumor suppressor is critically involved in antiviral defense mediated by intracellular IFN. PMID- 15752773 TI - Growth kinetics of SARS-coronavirus in Vero E6 cells. AB - Vero E6 cells are commonly used for in vitro studies of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and for antiviral evaluation purposes. A better understanding of the SARS-CoV growth kinetics in Vero E6 cells is crucial to help elucidate the mechanism of antiviral activity of selective antiviral agents. In this study, the growth kinetics of SARS-CoV in Vero E6 cells were studied by quantitation of intra- and extracellular viral RNA load as well as extracellular virus yield at different time points post infection. At 12h post-infection, the intracellular viral RNA load was 3x10(2) fold higher than at the time of infection, and the extracellular viral RNA load was increased with a factor of 2 x 10(3). Intracellular viral RNA levels started to rise at 6h post-infection. One hour later (at 7h post-infection), the levels of extracellular SARS-CoV RNA also began to rise. This was corroborated by the fact that infectious progeny SARS-CoV also first appeared in the supernatant between 6 and 7h post-infection. At 12h post-infection, SARS-CoV reached titers in the supernatant of 5.2 x 10(3) CCID(50)/ml. PMID- 15752774 TI - A novel mutation in the mitochondrial tRNA Asn gene associated with a lethal disease. AB - We describe a lethal mitochondrial disease in a 10-month-old child who presented with encephalomyopathy. Histochemical and electron microscopy examinations of skeletal muscle biopsy revealed abnormal mitochondria associated with a combined deficiency of complexes I and IV. After excluding mitochondrial DNA deletions and depletion, direct sequencing was used to screen for mutation in all transfer RNA (tRNA) genes. A T-to-C substitution at position 5693 in the tRNA(Asn) gene was found in blood and muscle. Microdissection of muscle biopsy and its analysis revealed the highest level of this mutation in cytochrome c oxidase (COX) negative fibres. We suggest that this novel mutation would affect the anticodon loop structure of the tRNA(Asn) and cause a fatal mitochondrial disease. PMID- 15752775 TI - Glucosamine induces rapid desensitization of glucose transport in isolated adipocytes by increasing GlcN-6-P levels. AB - We have examined the hypothesis that glucosamine (GlcN) can rapidly induce insulin resistance through an allosteric mechanism. When insulin-treated adipocytes were exposed to 2mM GlcN, glucose uptake was rapidly reduced by approximately 60% with a T(1/2) of 2 min. We also observed an increase in intracellular GlcN-6-P (at 5 min) from undetectable levels to approximately 260 nmol/g. Continued GlcN treatment resulted in additional accumulation of GlcN-6-P (>1200 nmol/g at 2h), but caused no further decrease in glucose uptake. Although the acute inhibitory action of GlcN could be completely reversed by removing extracellular GlcN, a slow and progressive decrease in insulin-stimulated glucose transport was observed with longer treatment times (T(1/2) of 45 min, 62% loss by 5h). From these data, we conclude that: (1) GlcN elevates intracellular GlcN-6-P levels within minutes, resulting in desensitization of the glucose transport system through allosteric inhibition of hexokinase; (2) prolonged treatment elevates GlcN-6-P to levels that cannot be effectively lowered by cell washing; and (3) residual levels of GlcN-6-P continue to allosterically inhibit glucose uptake, resulting in a slower rate of desensitization that is temporally similar to glucose-induced desensitization, but mechanistically different. PMID- 15752776 TI - LARGE2 facilitates the maturation of alpha-dystroglycan more effectively than LARGE. AB - The LARGE gene is thought to encode a putative glycosyltransferase because of its typical topology. However, no enzyme activity has been demonstrated yet, although the gene apparently supports the functional maturation of alpha-dystroglycan by glycosylation when it is transfected into cells. A novel homologous gene to LARGE was identified and named LARGE2. LARGE2 recombinant was co-expressed with alpha dystroglycan in human embryonic kidney 293T cells to determine its activity to support the maturation of alpha-dystroglycan. The alpha-dystroglycan co transfected with LARGE2 was more highly glycosylated than that co-transfected with LARGE. Pull-down experiments demonstrated binding activity of LARGE2 as well as LARGE toward alpha-dystroglycan. LARGE2 was found to support the maturation of alpha-dystroglycan more effectively than LARGE. Both of them are ubiquitously expressed in many tissues, except the brain where LARGE2 was not expressed at all. This compensatory function can explain the residual functionally glycosylated alpha-dystroglycan in a patient with MDC1D whose LARGE genes are congenitally null. PMID- 15752777 TI - Dendrimer modified magnetite nanoparticles for protein immobilization. AB - A cascading polyamidoamine (PAMAM) dendrimer was synthesized on the surface of magnetite nanoparticles to allow enhanced immobilization of bovine serum albumin (BSA). Characterization of the synthesis revealed exponential doubling of the surface amine from generations one through four starting with an amino silane initiator. Furthermore, transmission electron microscopy (TEM) revealed clear dispersion of the dendrimer-modified magnetite nanoparticles in methanol solution. The dendrimer-modified magnetite nanoparticles were used to carry out magnetic immobilization of BSA. BSA immobilizing efficiency increased with increasing generation from one to five and BSA binding amount of magnetite nanoparticles modified with G5 dendrimer was 7.7 times as much as that of magnetite nanoparticles modified with only aminosilane. There are two major factors that improve the BSA binding capacity of dendrimer-modified magnetite nanoparticles: one is that the increased surface amine can be conjugated to BSA by a chemical bond through glutaraldehyde; the other is that the available area has increased due to the repulsion of surface positive charge. PMID- 15752778 TI - Energy landscapes for adsorption of a protein-like HP chain as a function of native-state stability. AB - Dynamic Monte Carlo (DMC) simulations of the adsorption of simple protein-like chains are used to more clearly define the molecular basis for the dependence of adsorption thermodynamics on the stability of the unique lowest-energy "native state" conformation of the chain. Arai and Norde were among the first to show that proteins of low native-state stability strongly denature upon adsorption to weakly attractive sorbent surfaces, while relatively modest changes in conformation are observed in stable proteins under identical adsorption conditions. When the protein has a low native-state stability, favorable adsorption entropies are typically observed in such systems, leading to the general belief that the chain gains conformational entropy during adsorption through a net reduction in intramolecular interactions specific to the native state structure. Analysis of energy landscapes generated from our DMC simulation results show that a net loss in specific intramolecular interactions can lead to a positive delta(ads)S under certain adsorption conditions. However, the influence of chain conformation on delta(ads)S is found to correlate more directly with the manner in which the unique states of the system are distributed among the energy levels available to the adsorbed chain. Delta(ads)S is found to tend toward a maximum for adsorption processes described by thermally averaged energy landscapes in which the energy levels carrying the highest Boltzmann weights have a high degree of conformational degeneracy. This condition is met when the average interaction energy between the chain and the sorbent equals that between two hydrophobic segments of the chain. PMID- 15752779 TI - Modeling the mechanism involved during the sorption of methylene blue onto fly ash. AB - Batch sorption experiments were carried out to remove methylene blue from its aqueous solutions using fly ash as an adsorbent. Operating variables studied were initial dye concentration, fly ash mass, pH, and contact time. Maximum color removal was observed at a basic pH of 8. Equilibrium data were represented well by a Langmuir isotherm equation with a monolayer sorption capacity of 5.718 mg/g. Sorption data were fitted to both Lagergren first-order and pseudo-second-order kinetic models and the data were found to follow pseudo-second-order kinetics. Rate constants at different initial concentrations were estimated. The process mechanism was found to be complex, consisting of both surface adsorption and pore diffusion. The effective diffusion parameter D(i) values were estimated at different initial concentrations and the average value was determined to be 2.063 x 10(-9)cm2/s. Analysis of sorption data using a Boyd plot confirms the particle diffusion as the rate-limiting step for the dye concentration ranges studied in the present investigation (20 to 60 mg/L). PMID- 15752780 TI - Effect of Cr(VI) anions on adsorption and desorption behavior of Cu(II) in the colloidal systems of two authentic variable charge soils. AB - Heavy metals in wastes exist as multiple pollutants. The study of the interactions between multiple pollutants and soils should be of significance in practice. In the present study, the effect of chromate on adsorption and desorption behavior of Cu(II) in two variable charge soils was investigated, with the emphasis on the adsorption and desorption equilibria of Cu(II). The results showed that chromate can affect adsorption and desorption of Cu(II) in the colloidal systems of two variable charge soils. The extent of the effect was related to the initial concentrations of chromate and Cu(II), the system pH, and the nature of the soils. The presence of chromate led to an increase in the adsorption of Cu(II). For example, in the presence of 0.5, 0.8, 1.0, and 1.5 mmol L(-1) of chromate, for the rhodic ferralsol the adsorption of Cu(II) increased by 15.3, 18.0, 19.0, and 20.2%, respectively. For the hyperrhodic ferrasol, the corresponding figures were 11.9, 17.0, 20.3, and 26.1%, respectively. The presence of chromate also caused an increase in the desorption of Cu(II). For instance, in the presence of 0.5, 1.0, and 1.5 mmol L(-1) of chromate, the desorption for the rhodic ferralsol increased by 16.9, 27.5, and 34.1%, respectively. For the hyperrhodic ferralsol, the corresponding figures were 18.1, 35.6, and 51.4%, respectively. The increments of the adsorption and desorption increased with the increase in equilibrium concentration of Cu(II) in the solution. For instance, when the equilibrium concentrations were 0.5, 1.0, 1.5, and 2.0 mmol L(-1), the increments for the rhodic ferralsol were 2.5, 3.2, 3.3, and 3.0 mmol kg(-1), respectively. For the hyperrhodic ferralsol, the corresponding figures were 2.9, 3.5, 4.0, and 4.2 mmol kg(-1), respectively. The effect of chromate for the hyperrhodic ferralsol was greater than that for the rhodic ferralsol. This is caused by the difference in the content of iron oxides for the two soils. The increments of the adsorption and the desorption of Cu(II) increased with the rise in pH, reaching a maximum value, and then decreased. It can be assumed that the increment of the adsorption was caused by the change in surface charge of the soils induced by the adsorption of chromate and the cooperative adsorption of chromate adsorbed and Cu(II). The increase of electrostatically adsorbed Cu(II) was responsible for the increase in the desorption of Cu(II). PMID- 15752781 TI - Adsorption of Co2+, Ni2+, Cu2+ and Zn2+ from 0.5 M NaCl and major ion sea water on a mixture of delta-MnO2 and amorphous FeOOH. AB - The pH(pzc) values of several mechanical mixtures of amorphous hydrous oxides of iron (amorphous FeOOH) and manganese (delta-MnO2) have been determined using the solid addition method. While the pH(pzc) of delta-MnO2 remains almost unchanged, the corresponding value for amorphous FeOOH tends to increase with increased proportion of delta-MnO2 in the mixtures. The adsorption behavior of Co2+, Ni2+, Cu2+, and Zn2+ with respect to pH on a mechanical mixture of 70% delta-MnO2 and 30% amorphous FeOOH from 0.5 M NaCl and major ion sea water has been studied. Since delta-MnO2 is much more active adsorbent than amorphous FeOOH at pH below 6.5, the adsorption data on mixture have not only been normalized with respect to the mass of delta-MnO2 in the mixture, but also compared with adsorption data on delta-MnO2 alone. It is interesting to note that though each trace metal behaves in a different way from the other especially with respect to the nature of electrolyte medium, it is generally observed that the adsorption on the mixed oxide system is higher than that on delta-MnO2 alone under similar condition. It is also observed that adsorption in major ion sea water at a particular pH value is lower than in 0.5 M NaCl solution. PMID- 15752782 TI - Carbon-mineral adsorbents prepared by pyrolysis of waste materials in the presence of tetrachloromethane. AB - Natural bentonite spent in the process of plant oil bleaching was used as an initial material for preparation of carbon-mineral adsorbents. The spent bleaching earth was treated using four procedures: T (thermal treatment); H (hydrothermal treatment); C (thermal treatment with addition of CCl4 vapor); M (modification of porous structure). Raw bentonite, RB (raw bleaching earth), and carbon materials prepared using plant oil were compared. The physicochemical characteristics of the adsorbents were determined using different methods: nitrogen adsorption/desorption, XRD, TEM, and MS-TPD. Carbon-mineral adsorbents contain from 5.23 to 19.92% C (w/w) and carbon adsorbents include from 84.2 to 91.18% C (w/w). Parallel processes of organic substance carbonization, porous structure modification, sublimation or evaporation of metal chlorides, and removal of hydrogen chloride take place during pyrolysis of waste mineral materials in the CCl4 atmosphere. PMID- 15752783 TI - Decationization and dealumination of clinoptilolite tuff and ammonium exchange on acid-modified tuff. AB - The paper presents results of investigation of exchange of the clinoptilolite tuff cations with hydrogen ions from HCl solution of concentration 0.1 mmol cm( 3) and ammonium ions solutions of concentrations 0.0071 to 2.6 mmol cm(-3). Molal concentrations, x (mmol g(-1)) of cations exchanged in acid solution and in ammonium ions solutions were compared with molal concentrations of cations obtained by determination of the cation-exchange capacity of clinoptilolite tuff. The obtained results show that at ammonium ion concentrations lower than 0.1 mmol cm(-3), with regard to exchange capacity for particular ions, best exchanged are Na+ ions, followed by Mg2+ and Ca2+ ions, while exchange of K+ ions is the poorest (Na+ > Mg2+ > Ca2+ > K+). At ammonium concentrations from 0.2 to 1 mmol cm(-3) the order is Na+ > Ca2+ > Mg2+ > K+. At concentrations higher than 1 mmol cm(-3) the order is Na+ > Ca2+ > K+ > Mg2+. The results are a consequence of the uptake of hydrogen ions by zeolite samples in ammonium ions solutions at concentrations lower than 1 mmol cm(-3) and indicate the importance of Mg2+ (besides Na+ ions) for the exchange between clinoptilolite cations and H+ ions, in contrast to K+ ions, whose participation in the reaction with H+ ions is the lowest. During decationization of the clinoptilolite in acid solution, best exchanged are Na+, Mg2+, and Ca2+ ions, while exchange of K+ ions is the poorest. Due to poor exchange of K+ and H+ ions and good exchange of Na+, Mg2+, and Ca2+ ions, it is to be assumed that preservation of stability of the clinoptilolite structure is caused by K+ ions present in the channel C. Clinoptilolite is dissolved in the clinoptilolite A and B channels where Na+, Mg2+, and Ca2+ ions are present. On the acid-modified clinoptilolite samples, exchange of ammonium ions is poorer than on natural zeolite. The longer the contact time of the zeolite and acid solution, the worse ammonium ions exchange. It can be assumed that H+ ions exchanged with zeolite cations are consumed for solution of aluminum in the clinoptilolite structure; therefore the concentration of H+ ions as exchangeable cations decreases. In the ammonium ion solution at a concentration of 0.0065 mmol cm(-3), from the acid-modified zeolite samples, Al3+ ions are exchanged best, followed by Na+, Mg2+, Ca2+, and K+ ions. Further to the results, it is to be assumed that exchangeable Al3+ ions available from clinoptilolite dissolution are best exchanged with H+ ions in acid solution. PMID- 15752784 TI - Scale inhibition study by turbidity measurement. AB - The concept of a critical supersaturation ratio (CSSR) has been used to characterize the effectiveness of different types of scale inhibitors, inhibitor concentration, and precipitating solution pH in order to prevent the formation of barium sulfate scale. The scale inhibitors used in this work were aminotrimethylene phosphonic acid (ATMP), diethylenetriaminepentamethylene phosphonic acid (DTPMP), and phosphinopolycarboxylic acid polymer (PPCA). The CSSR at which barium sulfate precipitates was obtained as a function of time for different precipitation conditions and was used as an index to evaluate the effect of the precipitation conditions. The results showed that the CSSRs decrease with increasing elapsed time after mixing the precipitating solutions, but increases with increasing scale inhibitor concentration and solution pH. The CSSR varies linearly with the log of the scale inhibitor concentration and with the precipitating solution pH. A SEM analysis showed that the higher the scale inhibitor concentration and solution pH, the smaller and more spherical the BaSO4 precipitates. Analysis of the particle size distribution revealed that increasing the elapsed time, the scale inhibitor concentration, and precipitating solution pH, all produce a broader particle size distribution and a smaller mean diameter of the BaSO4 precipitates. DTPMP and PPCA were the most effective BaSO4 scale inhibitors per ionizable proton and the most effective on a concentration basis, respectively. PMID- 15752785 TI - Structural and adsorptive properties of Ba or Mg oxide modified zirconia. AB - Zirconia nanoparticles modified by barium oxide or magnesium oxide were synthesized by using a co-precipitation process followed by ethanol supercritical drying. The nanoparticles obtained were further calcined at 873 K. BET surface area, XRD, and TGA were used to characterize the prepared samples. Isotherms of N2 and CO2 adsorption on these modified zirconia nanoparticles were measured at various temperatures. Additions of BaO or MgO resulted in an increase in CO2 adsorption capacity of the modified zirconia particles. Results also show that BaO as a modifier is more effective than MgO in enhancing the CO2 adsorption capacity of zirconia. At 1 bar and 473 K, Ba modified zirconia adsorbs approximately 0.25 mmol/g of CO2. PMID- 15752786 TI - Determination of surface properties of iron hydroxide-coated alumina adsorbent prepared for removal of arsenic from drinking water. AB - A novel type adsorbent was prepared by in situ precipitation of Fe(OH)3 on the surface of activated Al2O3 as a support material. The iron content of the adsorbent was 0.31+/-0.003% m/m (56.1 mmol/g); its mechanical and chemical stability proved to be appropriate in solutions. The total capacity of the adsorbent was 0.12 mmol/g, and the pH of zero point of charge, pH(zpc) = 6.9+/ 0.3. Depending on the pH of solutions, the adsorbent can be used for binding of both anions and cations, if pH(eq) < pH(zpc) anions are sorbed on the surface of adsorbent (S) through [SOH2+] and [SOH] groups. A graphical method was used for the determination of pH(iep) (isoelectric points) of the adsorbent and values of pH(iep) = 6.1+/-0.3 for As(III) and pH(iep) = 8.0+/-0.3 for As(V) ions were found. The amount of surface charged groups (Q) was about zero within the a pH range of 6.5-8.6, due to the practically neutral surface formed on the adsorption of As(V) ions. At acidic pH (pH 4.7), Q = 0.19 mol/kg was obtained. The adsorption of arsenate and arsenite ions from solutions of 0.1-0.4 mmol/L was represented by Langmuir-type isotherms. A great advantage of the adsorbent is that it can be used in adsorption columns, and low waste technology for removal of arsenic from drinking water can be developed. PMID- 15752787 TI - Adsorption of methylene blue onto jute fiber carbon: kinetics and equilibrium studies. AB - Jute fiber obtained from the stem of a plant was used to prepare activated carbon using phosphoric acid. Feasibility of employing this jute fiber activated carbon (JFC) for the removal of Methylene blue (MB) from aqueous solution was investigated. The adsorption of MB on JFC has found to dependent on contact time, MB concentration and pH. Experimental result follows Langmuir isotherm model and the capacity was found to be 225.64 mg/g. The optimum pH for the MB removal was found to be 5-10. The kinetic data obtained at different concentrations have been analyzed using a pseudo-first-order, pseudo-second-order equation, intraparticle diffusion and Elovich equation. Among the kinetic models studied, the intraparticle diffusion was the best applicable model to describe the adsorption of MB onto JFC. PMID- 15752788 TI - Adsorption of benzoic acid onto high specific area activated carbon cloth. AB - The adsorption of benzoic acid from aqueous solution onto high area carbon cloth at different pH values has been studied. Over a period of 125 min the adsorption process was found to follow a first-order kinetics and the rate constants were determined for the adsorption of benzoic acid at pH 2.0, 3.7, 5.3, 9.1, and 11.0. The extents of adsorption and the percentage coverage of carbon cloth surfaces were calculated at 125 min of adsorption. Adsorption isotherms at pH values of 2.0, 3.7, and 11.0 were derived at 25 degrees C. Isotherm data were treated according to Langmuir and Freundlich equations and the parameters of these equations were evaluated by regression analysis. The fit of experimental isotherm data to both equations was good. It was found that both the adsorption rate and the extent of adsorption at 125 min were the highest at pH 3.7 and decreased at higher or lower pH values. The types of interactions governing in the adsorption processes are discussed considering the surface charge and the dissociation of benzoic acid at different pH values. PMID- 15752789 TI - Adsorption and desorption studies of a water soluble dye, Quinoline Yellow, using waste materials. AB - Bottom ash, a power plant waste, and de-oiled soya, an agricultural waste material, were employed for the removal and recovery of Quinoline Yellow, a water soluble dye. Characterization of adsorbent materials was made by their infrared and differential thermal analysis curves. Along with batch adsorption studies, which involve effect of pH, adsorbate concentration, sieve size, adsorbent dosage, contact time, temperature, etc., kinetic studies and column operations were also made to remove the dye from wastewater. On the basis of kinetic studies, specific rate constants involved in the processes were calculated and first-order adsorption kinetics was observed in both the cases. The paper also incorporates Langmuir and Freundlich adsorption isotherm models, which are used to calculate thermodynamic parameters and also to suggest a plausible mechanism of the ongoing adsorption processes. Fixed bed columns were prepared for both the adsorbents and bulk removal of the dye was achieved by eluting aqueous solution of the dye and saturation factor for both columns were evaluated. Dilute NaOH solution was then percolated through the exhausted columns to recover the adsorbed dye. PMID- 15752790 TI - Enzymatic degradation of model cellulose films. AB - Enzymatic degradation of model cellulose films prepared by a spin-coating technique was investigated by ellipsometry. The cellulose films were prior to degradation characterized by ellipsometry, contact angle measurements, ESCA (electron spectroscopy for chemical analysis) and AFM (atomic force microscopy). At enzyme addition to preformed cellulose films an initial adsorption was observed, which was followed by a total interfacial mass decrease due to enzymatic degradation of the cellulose films. The degradation rate was found to be constant during an extended time of hours, whereafter the degradation leveled off. In parallel to the decreased interfacial mass, the cellulose degradation resulted in a thinner and more dilute interfacial film. At long degradation times, however, there was an expansion of the cellulose film. The enzyme concentration affected the degradation rate significantly, with a faster degradation at a higher enzyme concentration. The effects of pH, temperature, ionic strength and stirring rate in the cuvette were also investigated. PMID- 15752791 TI - Combined study of X-ray reflectivity and atomic force microscopy on a surface grafted phospholipid monolayer on a solid. AB - We investigated the detailed structure of a surface-grafted phospholipid monolayer, which was polymerized in situ onto a methacryloyl-silanized solid surface. By the combined study of X-ray reflectivity and atomic force microscopy, the in situ polymerization step of the lipid molecules are sufficiently detailed to reveal the molecular structure of lipid molecules before and after in situ polymerization. From the data of the X-ray reflectivity, we confirmed that the in situ polymerization process produces a flat lipid monolayer structure and that the lipid monolayer is substantially grafted on a silanized surface by chemical bonding. After the polymerization and washing processes, the thickness of the head group was 9 angstroms and the thickness of the tail group was 21 angstroms. The surface morphology of the polymerized phospholipid monolayer obtained by the measurements of atomic force microscopy was consistent with the results of the X ray reflectivity. The cross-sectional analysis shows that the surface coverage of lipid molecules, which are chemically grafted onto a silanized surface, is approximately 89%. PMID- 15752792 TI - Biosensor signal amplification of vesicles functionalized with glycolipid for colorimetric detection of Escherichia coli. AB - A supramolecular assembly of phospholipid-polymerized diacetylene vesicles functionalized with glycolipid can provide a molecular recognition function. The Escherichia coli-glycolipid binding event leads to a visible color change from blue to red, readily seen with the naked eye and quantified by absorption spectroscopy. The biosensor signal is amplified through a suitable increase of phospholipid content in the mixed lipid vesicles and pH of aqueous solutions. PMID- 15752793 TI - Film formation from monodisperse acrylic lattices 1. Influence of concentration and layer thickness on particle ordering. AB - The influence of concentration and layer thickness on particle ordering in polymer latex films, both open and closed, has been studied by means of rheology, microscopy and turbidimetry. Monodisperse acrylic lattices were synthesized by semicontinuous emulsion polymerization. The lattices exhibited a distinct thixotropy above a certain concentration, which is attributed to crystallization. Microscopy revealed a three-layer structure and a dependence of crystal size and crystal packing on layer thickness and layer concentration. Turbidimetry (i.e., analysis of light transmission and interference) was used to study the progress of ordering in open and closed systems. In closed films crystallization proceeds faster at higher concentrations and in thinner films. Above a sufficiently high concentration no crystallization was observed. Furthermore, an induction time was found below certain concentrations. The drying of open latex films at temperatures below the minimum film formation temperature (MFFT) was shown to proceed through five distinct stages. The drying of open films at temperatures below the MFFT was studied by analysis of the turbidity of rewetted films. A more compact structure was found in thinner films. The structure in lattices is discussed as a result of long-range and short-range ordering. PMID- 15752794 TI - Film formation from monodisperse acrylic lattices 2. Influence of drying temperature on the film formation process. AB - The influence of drying temperature on the properties of latex films was investigated by gravimetry, turbidimetry (i.e., analysis of transmission spectra and interference), atomic force microscopy and measurement of water vapor permeability. Several pitfalls in the determination of water content of dried films that absorb water after being submerged in it have been proposed, such as fading boundaries, remaining water after drying and change of particle sizes. At moderately higher temperatures film formation is improved. This improvement follows from the formation of smoother film surfaces (AFM), lower water vapor permeabilities and smaller values for delta lambda(min). On the other hand, defects as cracks and channels also are created, especially at high temperatures. It appears, however, that these channels do not run from the one surface of a film to the other. PMID- 15752795 TI - Hofmeister effects on the colloidal stability of an IgG-coated polystyrene latex. AB - The effect of various ions related to the Hofmeister series (HS) on different properties of a cationic latex covered with a protein (IgG) is analyzed in this study. NaNO3, NH4NO3, and Ca(NO3)2 were used to compare the specificity of the cations, and NaCl, NaSCN, NaNO3, and Na2SO4, to compare the specificity of the anions. Two pH values, 4 and 10, were chosen to analyze the behavior of these ions acting as counter- and co-ions. At pH 4, the total surface charge is positive, whereas at pH 10 it is negative. Three different phenomena have been studied in the presence of these Hofmeister ions: (1) colloidal aggregation, (2) electrophoretic mobility, and (3) colloidal restabilization. The specific effect of the ions was clearly observed in all experiments, obtaining ion sequences ordered according to their specificity. The most important parameter for ion ordering was the sign of the charge of the colloidal particle. Positively charged particles displayed an ion order opposite that observed for negatively charged surfaces. Another influential factor was the hydrophobic/hydrophilic character of the particle surface. IgG-latex particle surfaces at pH 10 were more hydrophilic than those at pH 4. The SCN- ion had a peculiar specific effect on the phenomena studied (1)-(3) at pH 10. With respect to the restabilization studies at high ionic strengths, new interesting results were obtained. Whereas it is commonly known that cations may provoke colloidal restabilization in negative particles when they act as counterions, our experiments demonstrated that such restabilization is also possible with positively charged particles. Likewise, restabilization of negative surfaces induced by the specific effect of chaotropic anions (acting as co-ions) was also observed. PMID- 15752796 TI - Flocculation of colloidal clay by bacterial polysaccharides: effect of macromolecule charge and structure. AB - The molecular mechanism of montmorillonite flocculation by bacterial polysaccharides was investigated, with special emphasis on the effect of carboxylic charges in the macromolecules on the mechanisms of interaction with the clay surface. An indirect way to quantify the energy of interaction was used, by comparing the flocculation ability of variously acidic polysaccharides. Data on tensile strength of aggregates in diluted suspension were collected by timed size measurements in the domain 0.1-600 microm, using laser diffraction. The flow behavior of settled aggregates was studied by rheology measurements. Flocculation of colloidal clay suspension by polysaccharides requires cancelling of the electrostatic repulsions by salts, which allows approach of clay surfaces close enough to be bridged by adsorbing macromolecules. The amount of acidic charges of the polysaccharides, and especially their location in the molecular structure, governs the bridging mechanism and the resulting tensile strength of the aggregates. The exposure of carboxylate groups located on side chains strongly promotes flocculation. In turn, charges located on the backbone of the polysaccharide are less accessible to interaction, and the flocculation ability of such polysaccharides is lowered. Measurements at different pH indicate that adsorption of acidic polysaccharides occurs via electrostatic interactions on the amphoteric edge surface of clay platelets, whereas neutral polysaccharides rather adsorb via weak interactions. Increased tensile strength in diluted aggregates due to strong surface interactions results in proportionally increased viscosity of the concentrated aggregates. PMID- 15752797 TI - Correlation of the gradient diffusion coefficients of human lactoferrin with interparticle interactions validated by photon correlation spectroscopy. AB - The gradient diffusion coefficients of recombinant human lactoferrin, a glycoprotein that is of commercial interest, have been extensively investigated theoretically and experimentally. A theoretical prediction was developed to allow calculation of the thermodynamic coefficient in terms of the electrostatic repulsive forces, London-van der Waals forces, entropic forces and additional interactions of unknown source and determination of the hydrodynamic coefficient by a perturbation theory method. In addition, dilute limit calculations were carried out for both of them. The thermodynamic and hydrodynamic coefficients were incorporated in the generalised Stokes-Einstein equation to calculate the gradient diffusion coefficient. Dynamic light scattering experiments were also conducted under different physiochemical conditions in which the gradient diffusion coefficients and the mean sizes of the macromolecules were measured. It was shown that of the theoretical approaches studied, good quantitative agreement between theory and experiment was found. PMID- 15752798 TI - Chemical destabilization of oil-in-water emulsion by novel polymerized diethanolamines. AB - The main objective of this study was to synthesize novel demulsifiers for resolving oil-in-water emulsions. Diethanolamine polyethers are considered as a cationic polymer type. The study describes an improved synthesis of a series of diethanolamine polyethers via condensation of 3-7 or 9 mol of diethanolamine. The structure and the molecular weights of the major components in the reaction mixture were confirmed via IR and MS analyses. The demulsifiers were used for treatment of pollution in the refinery wastewater with or without FeCl3. The flocculation efficiency of the synthesized demulsifiers was determined by turbidity measurement of the treated and untreated O/W emulsion in the Cairo Oil Refinery Company. The critical flocculation concentration (CFC) and charge density of the synthesized demulsifiers were determined. Biodegradation of diethanolamine polyethers was measured in river water within 7-8 days. PMID- 15752799 TI - Investigation of the interfacial properties of water-in-diluted-bitumen emulsions using micropipette techniques. AB - The interfacial properties of water-in-diluted bitumen emulsions were studied using micropipette techniques. It was observed that, as bitumen concentration in the bulk phase (C0) increased, the interfacial tension on the water droplet surfaces decreased. In addition, there was a small effect on the interfacial tension when different solvent mixtures were used. Mixtures of toluene and heptane in different ratios were used as solvents for bitumen dilution. Crumpling of the interface was influenced by bitumen concentration and type of solvent. No crumpling was found for bitumen content less than 0.01% for all solvents used. Crumpling was observed at higher bitumen concentrations when deionized water (pH 5.4-5.6) was used. Setting "heptol[A]" to be the mixture of toluene and heptane, with the volume percent of toluene being A, the following were concluded. Crumpling disappeared at C0 > 1% and when heptol[100] was used, and also at C0 > 10% and when heptol[30] was used. Crumpling was strongly affected by the water pH. In the case of heptol[50], at a higher pH, the crumpling region that normally occurred at C0 > 0.01% disappeared. The micropipette technique proved to be useful in studying the interfacial properties of micrometer-sized emulsion drops. PMID- 15752800 TI - Preparation of ZnO nanoparticles in a reverse micellar system and their photoluminescence properties. AB - ZnO nanoparticles with spherical morphology and narrow size distribution were obtained by calcination of Zn(OH)2 nanoparticles, which were prepared in a polyethylene glycol mono-4-nonylphenyl ether (NP-5)/cyclohexane reverse micellar system and incorporated into polyurea (PUA) via an in situ polymerization of hexamethylene diisocyanate (HDI). The resulting ZnO nanoparticles demonstrated a near-UV emission and a green emission, the intensity ratio of which depended on calcination conditions. For the nanoparticles studied, the calcination atmosphere influenced remarkably the photoluminescence properties such as intensity ratio of the near-UV emission to green emission, rather than the size, morphology, and crystallinity of the ZnO nanoparticles. The green emission decreased by calcination in O2 flow but increased by calcination in N2 flow, as compared with the case calcined in air flow. This finding suggests that the green emission is enhanced with the increase of the number of oxygen vacancies of the ZnO nanoparticles and thus the photoluminescence properties of the nanoparticles were successfully controlled by the calcination condition, without changing the size and morphology. PMID- 15752801 TI - Recovery of nanoparticles produced in phosphatidylcholine-based template phases. AB - This paper focuses on the characterization and use of polymer-modified phosphatidylcholine (PC)/sodium dodecyl sulfate (SDS)-based inverse microemulsions as a template phase for BaSO4 nanoparticle formation. The area of the optically clear inverse microemulsion phase in the isooctane/hexanol/water/PC/SDS system is not significantly changed by adding polyelectrolytes, i.e., poly(diallyldimethylammonium chloride) (PDADMAC), or amphoteric copolymers of diallyldimethylammonium chloride and maleamid acid to the SDS-modified inverse microemulsion. Shear experiments show non-Newtonian flow behavior and oscillation experiments show a frequency-dependent viscosity increase (dilatant behavior) of the microemulsions. Small amounts of bulk water were identified by means of differential scanning calorimetry. One can conclude that the macromolecules are incorporated into the individual droplets, and polymer-filled microemulsions are formed. The polymer-filled microemulsions were used as a template phase for the synthesis of BaSO4 nanoparticles. After solvent evaporation the nanoparticles were redispersed in water and isooctane, respectively. The polymers incorporated into the microemulsion are involved in the redispersion process and influence the size and shape of the redispersed BaSO4 particles in a specific way. The crystallization process mainly depends on the type of solvent and the polymer component added. In the presence of the cationic polyelectrolyte PDADMAC the crystallization to larger cubic crystals is inhibited, and layers consisting of polymer-stabilized spherical nanoparticles of BaSO4 (6 nm in size) will be observed. PMID- 15752802 TI - Preparation of gold-, gold/silver-dendrimer nanocomposites in the presence of benzoin in ethanol by UV irradiation. AB - Gold- and gold/silver-dendrimer nanocomposites have been synthesized by UV irradiation of their salts dissolved in ethanol containing dendrimers. As dendrimers, poly(amidomaine) PAMAM dendrimers and poly(propyleneimine) PPI dendrimers of various generations were used. The photoreduction of their salts is greatly accelerated by using benzoin as a photoinitiator. The sizes of gold in the nanocomposites are affected by the concentration of benzoin as well as the concentration of dendrimers, but are hardly changed with the kind of dendrimers. For gold/silver-dendrimer nanocomposites, the absorption spectra of gold/silver nanoparticles in the nanocomposites are very similar to the theoretical spectra of gold/silver alloy nanoparticles, suggesting the formation of gold/silver alloy nanoparticles. From the comparison of TEM and DLS measurements, it is found that the metal-dendrimer nanocomposites consist of metal nanoparticles covering by dendrimer molecules. PMID- 15752803 TI - Improvement of red mud polymer-matrix nanocomposites by red mud surface treatment. AB - The effects of chemical treatments on red mud (RM) were investigated in terms of thermal stabilities of PMMA/RM and PVC/RM nanocomposites. N2/77 K adsorption behavior and contact angles were studied in the pore structures and surface energetics of RM, respectively. Thermal stabilities of the nanocomposites were investigated using a thermal mechanical analyzer (TMA) and thermogravimetric analysis (TGA). As a result, the acidically treated RM (ARM) had higher adsorption properties, including specific surface area, than untreated RM (VRM) or basically treated RM (BRM). A change in the structure of the ARM surface was due to hydrolysis or leaching a metal salt out of RM. Also, the electron acceptor (gamma(S)+, acid) of ARM and the electron donor (gamma(S)-, base) of BRM were increased in the development of acid and basic functional groups, respectively. PMMA/ARM nanocomposites had higher thermal stability and mechanical interfacial properties than PMMA/VRM or BRM nanocomposites. These results were due to the improvements of the dispersion properties and acid-base interfacial interactions of basic PMMA and ARM. In this work, although the dispersion properties of the BRM decreased, the thermal stabilities and mechanical interfacial properties of PVC/BRM nanocomposites increased, which could be attributed to improvement in the interfacial interactions between acidic PVC and BRM. PMID- 15752804 TI - Electrical double-layer effects on the deposition of zeolite A on surfaces. AB - Zeolite particles formed from an aluminosilicate solution possess a negative surface charge due to the substitution of aluminum atoms into a SiO4 tetrahedral structure making it difficult to form a continuous layer in solution. The particle interactions with surfaces and each other can be studied using the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. The interaction energy between zeolite-zeolite and zeolite-substrate on various materials can be estimated in this fashion. The zeolite LTA particles show a stronger repulsion interaction on all substrates and on the each other as compared to the ZSM-5 particles. This repulsive energy also increases as the particles size increases. This results in the formation of conglomerate in the solution rather than forming an adhered layer on the substrate. PMID- 15752805 TI - Covalent assembly of shortened multiwall carbon nanotubes on polyelectrolyte films and relevant electrochemistry study. AB - A significant and versatile approach was developed for perpendicularly aligning multiwall carbon nanotubes on diverse substrates suitable for layer-by-layer self assembly. The multiwall carbon nanotubes (s-MWNTs) used were shortened with oxidation under ultrasonic and functionalized with acyl chloride in thionyl chloride (SOCl2). The monolayer of s-MWNTs perpendicularly grafted to the substrate was obtained by dipping the polyelectrolyte modifying substrate into a tetrahydrofuran suspension of the functionalized s-MWNTs. The interaction proved to be stable and not liable to be affected by the ambience. Transmission electron microscopy and atomic force microscopy were used to examine the morphology of the MWNTs and s-MWNTs grafted on the substrates. Raman spectroscopy was applied to verify the existence of s-MWNTs for assembly, and Fourier transform infrared absorption spectra were used to investigate the interaction pattern between s MWNTs and polyelectrolyte. The electrochemistry properties of the monolayer of s MWNTs when the substrate was indium-tin oxide were studied. PMID- 15752806 TI - Effect of drying on the mesoporous structure of sol-gel derived silica with PPO PEO-PPO template block copolymer. AB - The effects of drying method on the pore structure of mesoporous silica were studied from the viewpoint of enhancing closed porosity in mesoporous silica. The mesoporous silica was prepared via a sol-gel process using polyethyleneoxide polypropyleneoxide-polyethyleneoxide (PEO-PPO-PEO) triblock copolymer (Pluronic P123) as the structure-directing template. The closed porosity was evaluated from the apparent mass density of the sample measured by a helium pycnometer. These mesoporous silicas were also characterized by transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and nitrogen adsorption. The drying method was shown to be responsible for the finally templated mesoporous structure of the silica. More rapid drying is more preferable for enhancing the closed porosity of the mesoporous silica. The closed pores were formed by immediate immobilization of copolymer molecular assemblies in the silica matrix due to the instant removal of the solvent and solidification at higher temperatures. The drying method, mainly affecting the drying rate, is highly influential on the finally replicated mesoporous structure in silica. PMID- 15752807 TI - Development of an ultrasonic technique for in situ investigating the properties of deposited protein during crossflow ultrafiltration. AB - Although an amount of research has reported that a flux minimum occurs at the isoionic/isoelectric points (pH 4.6-5.0) in the absence of salts in the ultrafiltration of bovine serum albumin (BSA), the real mechanism remains incompletely understood due to the lack of additional techniques in real time to detect the properties of deposited BSA (gel) layers formed during ultrafiltration (UF). An ultrasonic technique was developed as an analytical noninvasive tool to in situ investigate the properties of deposited BSA layers at pH 4.9 (isoionic or isoelectric point, IEP) and 6.9 during crossflow ultrafiltration. The membrane was a polysulfone (PSf) UF membrane with molecular weight cut-off (MWCO) 35 kDa. The feed used was 0.5 g/l BSA solution. Results show good correspondence between the ultrasonic signal responses and the development of BSA gel layers on the membranes. The deposit is thicker at pH 6.9 than at pH 4.9. However, the deposited gel layers are more compressible at pH 4.9 than at pH 6.9. The flux decline is mainly controlled by the density (packing) of the deposit layer. At pH 6.9, protein mainly deposits on the membrane surface. Around the isoelectric point, protein absorbs within and on the membranes. A functional relationship between acoustic signals and fouling resistance exists. The fouling resistance is mainly attributed to pore blocking or pore constriction. PMID- 15752808 TI - Effect of the intercalation conditions of a montmorillonite with octadecylamine. AB - Intercalation of montmorillonite with octadecylamine under several conditions is reported. Octadecylamine was protonated in situ with HCl to obtain octadecylammonium cations. Water and water/ethanol mixtures were used as reaction medium, and the ratios amine/clay and HCl/amine were also varied. Intercalation was successful when the amine/clay ratio was in the range 1-3 mmol/g; optimal results were obtained for an amine/clay ratio of 2.0 mmol/g. For a given amine/clay ratio, the HCl/amine ratio also influences the intercalation. Basal spacings of the intercalated solids were between 13.4-36.7 angstroms. The amount of organic matter incorporated into the solids also varied widely; up to 40 wt% is fixed. Specific surface area is very low in all the intercalated solids because of the blockage of the clay porosity by the organic molecules. Co intercalation of octadecylammonium and of the inorganic polycation [Al13O4(OH)24(H2O)12]7+ was also considered, giving rise to intercalated solids with basal spacings between 17 and 22 A, also with a high fixation of organic matter. PMID- 15752809 TI - Observation and modelling of clamshell droplets on vertical fibres subjected to gravitational and drag forces. AB - Extensive experimental investigation of the wetting processes of fibre-liquid systems during air filtration (when drag and gravitational forces are acting) has shown many important features, including droplet extension, oscillatory motion, and detachment of drops from fibres as airflow velocity increases, and also movement or flow of droplets along fibres. A detailed experimental study of the processes was conducted using stainless steel filter fibres and H2O aerosol, which coalesce on the fibre to form clamshell droplets. The droplets were predominantly observed in the Reynolds transition flow region, since this is the region where most of the above features occur. The droplet oscillation is believed to be induced by the onset of the transition from laminar to turbulent flow as the increasing droplet size increases Reynolds number for the flow around the droplet. Two-dimensional flow in this region is usually modelled using the classical Karman vortex street, however there exist no 3D equivalents. Therefore to model such oscillation it was necessary to create a new conceptual model to account for the forces both inducing and preventing such oscillation. The agreement between the model and experimental results is very good for both the radial and transverse oscillations. PMID- 15752810 TI - Correlation between surface and bulk structures of alcohol-water mixtures. AB - Adsorption isotherms of binary aqueous solutions of methanol, ethanol, 1 propanol, 2-propanol, tert-butanol, and 1-butanol are demonstrated, being calculated by using the Gibbs adsorption equation with experimental data of surface tension and vapor pressure found in the literature. For all of the alcohol-water mixtures, the maximum value in the adsorption isotherm, namely, the maximum surface excess is about that expected for the formation of a monolayer. Furthermore, the composition of the mixture for the maximum surface excess coincides with that corresponding to the minimum in the excess partial molar volume of the solutes. These results indicate that the hydrophobic hydration in bulk induces the surface excess of the alcohols and after a monolayer is formed, the hydrophobic hydration itself is no longer retained. PMID- 15752811 TI - Capillary rise kinetics of some building materials. AB - The presence of water in masonry is one of the main factors in deterioration. Capillary rise is the most usual mechanism of water penetration into building materials. In this study the kinetics of the capillary rise phenomenon was studied for various building materials: four stones, two bricks, and six plasters. A first-order kinetic model was proposed, in which the equilibrium moisture height derived from Darcy law. The capillary height time constant found to be strongly affected by the material characteristics. Moreover, the capillary height time constant can be predicted if the average pore radius of the materials is known. PMID- 15752812 TI - Wetting by simple room-temperature polymer melts: deviations from Newtonian behavior. AB - The hydrodynamics near moving contact lines of two room-temperature polymer melts, polyisobutylene (PIB) and polystyrene (PS), are different from those of a third polymer melt, polydimethylsiloxane (PDMS). While all three fluids exhibit Newtonian behavior in rotational rheological measurements, a model of the hydrodynamics near moving contact lines which assumes Newtonian behavior of the fluid accurately describes the interface shape of a variety of PDMS fluids but fails to describe the interface deformation by viscous forces in PIB and PS. The magnitude of the deviations from the model and the distance along the liquid vapor interface over which they are seen increase with increasing capillary number. We conclude that the wetting behaviors of PIB and PS are influenced by weak elasticity in these low molecular weight melts and that dynamic wetting is more sensitive to this elasticity than standard rheometric techniques. PMID- 15752813 TI - Swelling-shrinkage measurements of bentonite using coupled environmental scanning electron microscopy and digital image analysis. AB - The swelling clays have been proposed as engineered barriers in geological disposal systems for waste because these materials are assumed to build a better impermeable zone around wastes by swelling. However, the swelling potential of soils is also considered a prevalent cause of damage to buildings and constructions. For these reasons, it is fundamental to investigate the physicochemical and mechanical behavior of swelling clays. In the current study, the swelling-shrinkage potential (aggregates scale) was estimated using an environmental scanning electron microscope (ESEM) coupled with a digital image analysis (DIA) program (Visilog). In fact, the isolated aggregates of raw and cation-exchanged bentonite were directly observed at different relative humidities in an ESEM chamber. Then the "Visilog" software was used to estimate the percent augmentation of the aggregate surface as a function of time and as a function of relative humidity. This estimation allows for the calculation of the swelling-shrinkage potential (%) of bentonite. Finally, a kinetic model of first order was tested to fit the kinetic experimental data of swelling-shrinkage potential. The results show that ESEM-DIA coupling can be a powerful method of estimating the swelling-shrinkage potential of expansive clays. In addition, the exponential models fit well with the kinetic experimental data. PMID- 15752814 TI - Hygroscopic growth of self-assembled layered surfactant molecules at the interface between air and organic salts. AB - We report here the self-assembly of surfactant molecules at the interface of air and the hygroscopic quaternary ammonium salt tetrabutylammonium acetate (TBAAc). Homogeneously dissolved surfactant molecules at 100 degrees C self-assemble upon contacting air due to high moisture adsorption by the organic salt when cooling down. Highly ordered lamellar phases with different lattice spacings have been observed when surfactants with various lengths of alkyl chains were used. C(n)TMAB/TBAAc systems showed all-trans conformation of interior methylene carbons and interdigited bilayers with an average CH2 increment of 0.119 nm, while C(n)NH2/TBAAc systems showed trans/gauche mixed conformations of interior methylene carbons and bilayers with an average CH2 increment of 0.247 nm. C(n)NH2s in C(n)NH2/TBAAc formed bilayers through water-mediated intermolecular hydrogen bonds with a water layer thickness of 0.51-0.61 nm. In C(n)TAB/TBAAc, as the head group of C(n)TAB is bigger, the interdigited bilayer thickness (d spacing) is smaller, because the bigger head groups accommodate enough space for alkyl tails to come in between them. PMID- 15752815 TI - Effect of head group polarity and spacer chain length on the aggregation properties of gemini surfactants in an aquatic environment. AB - The aggregation behavior of cationic gemini surfactants with respect to variation in head group polarity and spacer length is studied through conductance, surface tension, viscosity, and small-angle neutron-scattering (SANS) measurements. The critical micellar concentration (cmc), average degree of micelle ionization (beta(ave)), minimum area per molecule of surfactant at the air-water interface (A(min)), surface excess concentration (gamma(max)), and Gibb's free energy of micellization (delta G(mic)) of the surfactants were determined from conductance and surface tension data. The aggregation numbers (N), dimensions of micelles (b/a), effective fractional charge per monomer (alpha), and hydration of micelles (h(E)) were determined from SANS and viscosity data, respectively. The increasing head group polarity of gemini surfactant with spacer chain length of 4 methylene units promotes micellar growth, leading to a decrease in cmc, beta(ave), and delta G(mic) and an increase in N and b/a. This is well supported by the observed increase in hydration (h(E)) of micelles with increase in aggregation number (N) and dimension (b/a) of micelle. PMID- 15752816 TI - Self-assembly and influence of the organic counterion in the ternary systems dodecylamine/acrylic acid/water and dodecylamine/methacrylic acid/water. AB - Due to complete proton transfer from the acid to the amine, a reaction between an equimolar mixture of dodecylamine and (meth)acrylic acid leads to the formation of dodecylammonium (meth)acrylate. The latter can be considered as a surfactant with a polymerizable organic counterion. The ternary phase diagrams of the two systems dodecylamine/acrylic acid/water and dodecylamine/methacrylic acid/water are described. Both systems can form isotropic solutions and lyotropic liquid crystalline lamellar phases. Moreover, the system with the methacrylate counterion can also form a cubic phase in the water-rich part of the phase diagram. The difference in the self-organization observed for the two systems is explained by the greater bulkiness and hydrophobicity of the methacrylate. Whereas the acrylate counterion behaves rather like a classic inorganic counterion, the methacrylate counterion resides in the outermost part of the aggregates, giving rise to a change in the surface curvature. PMID- 15752817 TI - Mixing behavior of fluorinated and hydrogenated gemini surfactants at the air water interface. AB - Mixing behavior of hydrogenated and fluorinated cationic gemini surfactants was studied at the air-water interface by Brewster angle microscopy and pi-A isotherm curves. In the bulk, these two molecules did not mix and showed phase separation. At the air-water interface, if a monolayer was formed by separate deposition of the two solutions, they formed separate domains, and the compression occurred in two steps: first the domains with hydrogenated gemini surfactant were compressed until they showed collapse; then the domains with fluorinated gemini surfactant were compressed. If the two solutions were mixed before the deposition, they remained mixed upon compression; on the other hand, separate domains under separate deposition were shown to mix if the subphase was heated. PMID- 15752818 TI - Two-fluid electroosmotic flow in microchannels. AB - This paper presents a mathematical model to describe a two-fluid electroosmotic pumping technique, in which an electrically non-conducting fluid is delivered by the interfacial viscous force of a conducting fluid; the latter is driven by electroosmosis. The electrical potential in the conducting fluid and the analytical solution of the steady two-fluid electroosmotic stratified flow in a rectangular microchannel was presented by assuming a planar interface between the two immiscible fluids. The effects of viscosity ratio, hold-up, concentration, and interfacial zeta potential are analyzed to show the potential feasibility of this technique. PMID- 15752819 TI - A new and simpler approach for the solution of the electrostatic potential differential equation. Enhanced solution for planar, cylindrical and annular geometries. AB - The motivation of the study performed in this project is focused on deriving a more effective, accurate, and mathematically friendly solution for the prediction of the electrostatic potential, commonly used on electrokinetic research and its related applications. In this contribution, based on the Debye-Huckel approximation, a new solution strategy for the differential equations of the electrostatic potential is proposed. In fact, a simple predictor-corrector calculation is developed to achieve more accurate predictions of electrostatic potential profiles. Furthermore, in this study the authors introduce the correction function f(AO) to the inverse Debye length, lambda. The f(AO) function improves the Debye-Huckel approximation and it is a recursive function of the electrical potential. Once the inverse Debye length, lambda, has been corrected by the f(AO) function and introduced in the simplified solution of the Poisson Boltzmann equation (i.e., the linear approximation, due to Debye and Huckel), the electrostatic potential outcome little differs from the numerical solution of the complete (nonlinear) differential equation. This new approach embraces different geometries of interest, such as planar, cylindrical, and annular, with excellent results in all the cases and for a wide range of electrostatic potential values. This new predicting semi-analytical technique can be a useful tool on electrical field applications such as the separation of a mixture of macromolecules and the removal of contaminants in soil cleaning processes. Illustrative results are presented for the geometries identified above. PMID- 15752820 TI - A new approach for analyzing particle motion near an interface using total internal reflection microscopy. AB - A new method was developed for analyzing the normal motion of a single colloidal particle near an interface. The optical technique of total internal reflection microscopy (TIRM) was used to determine the distribution of vertical displacements of a particle from a specific starting position as a function of time. At very small displacement times, the displacements are normally distributed with a variance that is proportional to the diffusion coefficient times the displacement time. The change in the diffusion coefficient with separation distance between the particle and plate was found to match that predicted by Brenner (Chem. Eng. Sci. 16 (1961) 242). As the sampling time becomes very large, the variance reaches a constant value determined strictly by the shape of the local potential energy profile holding the particle. A major advantage of this approach, relative to other measurement methods, is that the particle's spatially variant diffusion coefficient can be determined without any knowledge of the forces acting on the particle. PMID- 15752821 TI - Drag of a dispersion of nonhomogeneously structured flocs in a flow field. AB - The influence of floc structure and floc concentration on the drag acting on a floc is investigated theoretically. A two-layer model is adopted to describe floc structure, and a cell model is used to simulate a floc dispersion. The influences of the key parameters of the problem under consideration, including floc concentration, Reynolds number, the ratio (permeability of outer layer/permeability of inner layer), and the ratio (thickness of outer layer/thickness of inner layer), on the drag coefficient are discussed. We show that the more heterogeneous the floc structure is, the greater the drag and the more significant the deviation of curve of variation of drag coefficient against Reynolds number from a Stokes-law-like relation. The drag on a floc declines with the decrease in floc concentration, and, due to the convective flow of the fluid, the distortion of streamlines surrounding a floc becomes more serious and the deviation of the variation of the curve of drag against Reynolds number from a Stokes-law-like relation is more significant. PMID- 15752822 TI - Thermophoresis in liquids: a molecular dynamics simulation study. AB - Thermophoresis in liquids is studied by molecular dynamics simulation (MD). A theory is developed that divides the problem in the way consistent with the characteristic scales. MD is then conducted to obtain the solution of each problem, which is to be all combined for macroscopic predictions. It is shown that when the temperature gradient is applied to the nonconducting liquid bath that contains neutral particles, there occurs a pressure gradient tangential to the particle surface at the particle-liquid interface. This may induce the flow in the interfacial region and eventually the particle to move. This applies to the material system that interacts through van der Waals forces and may be a general source of the thermophoresis phenomenon in liquids. The particle velocity is linearly proportional to the temperature gradient. And, in a large part of the given temperature range, the particle motion is in the direction toward the cold end and decreases with respect to the temperature. It is also shown that the particle velocity decreases or even reverses its sign in the lowest limit of the temperature range or with a particle of relatively weak molecular interactions with the liquid. The characteristics of the phenomenon are analyzed in molecular details. PMID- 15752823 TI - Temperature-induced sedimentation to dispersion of ionic vesicles. AB - We present a temperature-induced sedimentation/dispersion transition of ionic vesicles in the system of alkyldimethylamine oxide hemihydrochloride (CnDMAO.1/2HCl) with a hydrocarbon chain length of 12-16 (n = 12, 14, and 16) and sodium 2-naphthalenesulfonate (NaNphS). The temperature-sensitive sedimentation/dispersion of ionic vesicles took place around a temperature of 50 degrees C, which was weakly dependent on the alkyl chain length. The combined effect of the thermally induced dissociation of the counterions from the vesicle and a hydrogen bonding between the nonionic and the cationic head groups is likely to be responsible for this unique behavior. PMID- 15752824 TI - A novel nanogold multilayer constructed by Langmuir-Blodgett and self-assembly techniques. AB - A multilayer comprising nanogold particles and organic molecules was constructed by self-assembly of the small organic molecule 1-phenyl-5-mercaptotetrazole onto the octadecylamine-protected nanogold monolayer in the course of transferring the monolayer, layer by layer, from a Langmuir-Blodgett trough to solid substrates. The structure and properties of the constructed multilayer were characterized by TEM images and ATR-IR, XPS, and UV-vis spectra. It is evidenced for the first time that the combination of Langmuir-Blodgett and self-assembly techniques provides a new way to organize functional nanoparticles such as organic-nanogold complexes. PMID- 15752825 TI - Enzymes in the cavity of hollow silica nanoparticles. AB - Due to limitations of the existing preparative methods of hollow nanoparticles by either heating at high temperature (>600 degrees C) or by using strong acid, alkali, or an organic solvent, it was not possible up till now to encapsulate any sensitive organic molecule like enzyme or others inside the cavity of hollow nanoparticles. We have demonstrated a much softer method of preparing hollow silica nanoparticles with horseradish peroxidase (HRP) inside the cavity by synthesizing HRP-doped core-shell silica-coated silver chloride nanoparticles and finally leaching out silver chloride with dilute ammonia at low temperatures. TEM pictures showed the hollow cavity inside the nanoparticles. The enzyme entrapped in these particles was active. The turnover number of HRP entrapped into these hollow particles and dispersed in aqueous buffer (pH 7.2) (k(cat) = 2.56 x 10(6) s(-1)) was found to be less than that of free enzyme in aqueous buffer (k(cat) = 6.133 x 10(7) s(-1)) but higher than that of HRP entrapped in solid-core silica nanoparticles and dispersed in aqueous buffer (k(cat) = 1.05 x 10(5) s(-1)). The result showed that hollow nanoparticles could be prepared using soft chemical methods and sensitive chemicals like active enzyme could be entrapped in the cavities and it retains its activity. PMID- 15752826 TI - Vaccines and unexpected observations: flaws or cause for concern? PMID- 15752827 TI - Hepatitis B virus genotypes. AB - Eight genotypes of hepatitis B virus (A-H) are currently recognized, and subgenotypes have recently been described in four of these genotypes (A, B, C and F). The genotypes show a distinct geographical distribution between and even within regions, and are proving to be an invaluable tool in tracing the molecular evolution and patterns and modes of spread of hepatitis B virus. Structural and functional differences between genotypes can influence the severity, course and likelihood of complications, and response to treatment of hepatitis B virus infection and possibly vaccination against the virus. Although the number of studies on these genotypes has increased dramatically during recent years, much remains to be learnt about their full implications. PMID- 15752828 TI - Interchangeability and tolerability of a virosomal and an aluminum-adsorbed hepatitis A vaccine. AB - The interchangeability of virosomal (Epaxal) and aluminum-adsorbed (Havrix 1440) hepatitis A virus (HAV) vaccines was studied in 111 healthy adults who were vaccinated in a randomized, single-blind, crossover clinical trial. Anti-HAV antibody titers were measured at days 0 (first dose), 14, and 28, and months 3, 6, 12 (second dose), 13, 24, 36, 48, 60 and 72. Most subjects (>95%) had sero converted 14 days after the first dose of either vaccine. The second dose with either vaccine induced a high antibody response in all vaccines, irrespective of the type of vaccine administered as the first dose. Although both vaccines were well tolerated, the incidence of local adverse events (in particular pain) was significantly lower in subjects receiving the virosomal vaccine. Six-year follow up data did not reveal any significant differences between the vaccination groups. PMID- 15752829 TI - Prime-boost vaccine regimen confers protective immunity to human-derived enterotoxigenic Escherichia coli. AB - Development of effective vaccines against diarrhea caused by enterotoxigenic Escherichia coli (ETEC) strains is still a priority for those living at or traveling to endemic regions. In this work, we evaluated the protective role of an anti-ETEC vaccine regimen based on parenteral priming with a DNA vaccine, pRECFA, followed by oral boosting with a recombinant attenuated Salmonella Typhimurium vaccine strain, HG3, both encoding the same antigen, the structural subunit (CfaB) of the ETEC CFA/I fimbriae. The DNA-priming Salmonella-boosting protocol enhanced both murine anti-CfaB serum IgG and fecal IgA antibody responses and increased the ability of serum antibodies to inhibit the adhesive properties of the CFA/I fimbriae expressed by live bacteria, as compared to mice immunized with only one vaccine type. Addition of a mucosal adjuvant (LTR192G) to the Salmonella vaccine strain further enhanced the synergic effects of the vaccine regimen on the induced CfaB-specific antibody responses. DBA/2 dams submitted to the prime-boost regimen transferred complete passive protection to suckling neonates challenged with a virulent ETEC strain. Detection of milk anti CfaB IgA antibodies and protection conferred by vaccinated dams to neonates born from non-vaccinated dams indicated that secretion of antigen-specific IgA is the immune response induced by the protective vaccine regimen. These results demonstrate that priming with a DNA vaccine and boosting with a Salmonella strain enhances both quantitatively and qualitatively the antibody responses to the CfaB antigen and represents an alternative for either active or passive immunization approach to ETEC-associated diarrhea. PMID- 15752830 TI - T-cell triggering by CD3- and CD28-binding molecules linked to a human virus modified tumor cell vaccine. AB - The aim was to develop T cell costimulatory molecules that are broadly applicable to augment anti-tumor immune responses upon application of a virus-modified tumor vaccine to cancer patients. We generated recombinant bispecific single-chain antibodies with one specificity directed against the CD3 or the CD28 antigen on human T cells and the other against the viral target molecule hemagglutinin neuraminidase (HN) of Newcastle Disease Virus (NDV). By re-directing unstimulated primary human T cells against HN-expressing NDV-infected tumor cells, the bispecific molecule bsHN-CD3 cross-linked effector and target cells and rapidly induced cytotoxicity at nanomolar concentrations. The bsHN-CD28 molecule exerted T cell co-stimulatory function. Maximal T cell activation was achieved with tumor cells infected by NDV and modified with both new stimulatory molecules. This was revealed by T cell proliferation, upregulation of CD69 and CD25 and by release of cytokines, interferons and chemokines. The new molecules combine high-effectivity with specificity and safety. PMID- 15752832 TI - Reactogenicity profile of a combined hepatitis A and B vaccine in clinical practice: a naturalistic study in adult travellers. AB - A prospective observational naturalistic study was conducted to assess the reactogenicity of the combined hepatitis A and hepatitis B (HAB) vaccine in a real-life setting. All healthy candidates for HAB vaccination attending an adult vaccination centre between October 1998 and February 2000 were invited to participate in the study. A follow-up diary card was provided to subjects to record local and general symptoms during a 4-day follow-up. Intensity was graded from 1 to 3. Redness was recorded as presence or absence. Fever was defined as axillar temperature > or =37.5 degrees C and grade 3 >39.0 degrees C. For all other symptoms, grade 3 was defined as an adverse reaction preventing normal everyday activities; 998 subjects (74% females), mean age (+/-S.D.) of 23 years (+/-4.5) (range: 11-54 years) agreed to participate. At first immunization 92% were <30 years old. Grade 3 pain and swelling was recorded in 1.2% and 0.3% of local symptom sheets completed, respectively; 438 subjects received the HAB vaccine alone (group 1) whereas 560 received at least one concomitant vaccine (group 2). In 45%, 27%, 18% and 10% of subjects the HAB vaccine was coadministered with 1, 2, 3 or 4 to 6 vaccines (mainly Td adult-type, typhoid, MMR and IPV vaccine). Grade 3 pain and swelling were recorded in 1.2% & 0.3% of symptom sheets (SS), respectively. In group 1, any fever and grade 3 fever was recorded in 3.5% and 0.1% of SS. Group 1 versus 2 had a lower risk for any fatigue (p=0.0002; OR=0.617) and any malaise (p=0.0076; OR=0.693) but not for grade 3 symptoms. In conclusion, our study showed that the HAB vaccine is well tolerated in adults either alone or coadministered with other vaccines in the routine clinical practice. PMID- 15752831 TI - Long lived multi-isotype anti-HIV antibody responses following a prime-double boost immunization strategy. AB - Despite decades of work, an effective HIV vaccine remains elusive. In an effort to elicit protective immunity, investigators have sought to define vaccines able to elicit durable HIV-specific B-cell and T-cell activities. Additionally, vaccines are sought which can induce antibodies of a variety of isotypes, as each isotype possesses unique attributes in terms of opsonization, Fc receptor binding capacity, complement fixation and location. One prominent new vaccine strategy, applied to numerous distinct antigenic systems is the prime boost-regimen, with DNA, vaccinia virus (VV), and/or purified recombinant protein. To examine the durability, location and isotype distribution of responses induced by prime-boost regimens, we tested successive immunizations with DNA, VV and protein (D-V-P), comparing three forms of protein inoculations: (i) purified protein administered intramuscularly with complete Freunds adjuvant, (ii) purified protein administered intranasally, and (iii) purified protein conjugated to oxidized mannan, administered intranasally. We found that all three protocols elicited serum antibodies of multiple isotypes, with serum IgA being most prominent among mice immunized with mannan-conjugated protein. All D-V-P protocols, regardless of protein form or route, also elicited antibody responses at mucosal surfaces. In bronchoalveolar lavage, a tendency toward IgA production was again most prominent in mice boosted with the protein-mannan conjugate. Both B-cell and T-cell responses were sustained for more than 1 year post-immunization following each form of vaccination. Contemporaneous with long-lasting serum and mucosal antibodies were antibody forming cells in the bone marrow of primed animals. Results highlight the D-V-P vaccination strategy as a promising approach for attaining durable, multi-isotype B-cell and T-cell activities toward HIV. PMID- 15752833 TI - An analytical framework for immunization programs in Canada. AB - Recent years have seen an increase in the number of new vaccines available on the Canadian market, and increasing divergence in provincial and territorial immunization programs as jurisdictions must choose among available health interventions with limited funding. We present an analytical framework, which we have developed to assist in the analysis and comparison of potential immunization programs. The framework includes 58 criteria classified into 13 categories, including the burden of disease, vaccine characteristics and immunization strategy, cost-effectiveness, acceptability, feasibility, and evaluability of program, research questions, equity, ethical, legal and political considerations. To date this framework has been utilized in a variety of different contexts, such as to structure expert presentations and reports and to examine the degree of consensus and divergence among experts, and to establish priorities. It can be transformed for a variety of other uses such as educating health professionals and the general public about immunization. PMID- 15752834 TI - Aerosol-, but not intradermal-immunization with the live vaccine strain of Francisella tularensis protects mice against subsequent aerosol challenge with a highly virulent type A strain of the pathogen by an alphabeta T cell- and interferon gamma- dependent mechanism. AB - Francisella tularensis is an extremely virulent facultative intracellular bacterial pathogen of many mammalian species including mice and humans in which it causes a spectrum of disease collectively called tularemia. In humans, intradermal or inhaled inocula of 10cfu or less of the most virulent strains of the pathogen are sufficient to cause severe infection and possible death; in mice similar inocula are routinely lethal. An attenuated live vaccine strain, F. tularensis LVS, was developed almost 50 years ago, and remains the sole prophylactic against virulent strains of the pathogen. Using F. tularensis LVS as a model vaccine, we recently showed that it was possible to systemically immunize various mouse strains and protect them against subsequent massive (2000 cfu) intradermal (i.d.) challenge, but not against low dose (approximately 10 cfu) aerosol challenge, with virulent strains of the pathogen. This is troubling because the latter route is considered an important means of deliberately disseminating F. tularensis in a bioterrorist attack. Others have previously shown that administering LVS to humans, guinea pigs and monkeys as an aerosol enhanced protection against subsequent aerosol challenge with virulent F. tularensis. In the present study, we show the same phenomenon in BALB/c and C3H/HeN mice. In this model, interferon gamma (IFNgamma) and CD4+ and CD8+ T cells are essential for the expression of anti-Francisella immunity in the lungs. Combined this immune response operates by limiting dissemination of the pathogen to susceptible internal organs. Further, understanding of how inhaled LVS elicits local cell-mediated protective immunity will be critical for devising improved vaccines against pulmonary tularemia. PMID- 15752835 TI - Confidence in vaccination: a parent model. AB - Although vaccination has been heralded as one of the 10 greatest public health achievements, how parents differ in their views about vaccination is not well understood. A deeper understanding of these attitudes and beliefs may improve the effectiveness of vaccine communications. In this mailed survey of U.S. parents in January 2001 (return response rate 49%), parental confidence in vaccination was very high, although there was significant variation among parents. Using multivariate analyses to group and profile parents, 90% of parents (n=1820) were classified into one of four distinct parent groups: (1) "Vaccine Believer" parents who were convinced of the benefit of vaccination; (2) "Cautious" parents noteworthy for a high emotional investment in their child; (3) "Relaxed" parents characterized by a less involved parenting style and some skepticism about vaccines; and (4) "Unconvinced" parents distinguished by their distrust of vaccinations and vaccination policy. These findings suggest that messages that are customized to parents' attitudes and beliefs may improve their understanding and acceptance of vaccination. PMID- 15752836 TI - Incidence of vaccine preventable pneumococcal invasive infections and blood culture practices in Italy. AB - In order to estimate the incidence of invasive pneumococcal diseases (IPD) and the amount of vaccine preventable serious infections, a 1-year population-based surveillance was undertaken in two comparable Italian regions (Piemonte and Puglia, representing 14% of the Italian population) prospectively collecting data and strains from all the hospital microbiological laboratories. A retrospective analysis of hospital discharge records, matched with the laboratory database, was also undertaken in nine hospitals in these two regions to determine the frequency of use of blood cultures and its impact on IPD incidence estimate. For children under 2 years of age, the incidence rates of IPD were 11.3 per 100,000 and 5.9 per 100,000 in Piemonte and in Puglia, respectively; for subjects 65 years of age and over the incidence rates were 5.7 per 100,000 and 0.2 per 100,000, in the two regions, respectively. The number of blood cultures performed was six times higher in Piemonte than that in Puglia. About 96% of isolates from IPD patients, aged 65 years and over, belonged to serogroups included in the 23-valent polysaccharide vaccine, whereas about 79% of strains isolated from patients under 5 years of age were related to serotypes included in the 7-valent conjugate vaccine. The estimate of the incidence of IPD is affected greatly by the different attitudes in performing blood cultures, especially in older patients. In Italy, bacteriological culture procedures should be undertaken more frequently to provide decision-makers with reliable estimates of serious vaccine preventable conditions. PMID- 15752837 TI - Production of yellow fever 17DD vaccine virus in primary culture of chicken embryo fibroblasts: yields, thermo and genetic stability, attenuation and immunogenicity. AB - While a good vaccine against yellow fever (YF) virus has been available for decades, the basic technology for the production of YF vaccine in chicken embryos has remained substantially unchanged since the 1940s. Here we describe the highly efficient and economic production of the 17DD strain of YF virus in chicken embryo fibroblast (CEF) cell cultures with viral titers ranging from 6.3 to 6.7 log10PFU/mL. Thermostability of two different formulations (5 and 50-dose vials) of the CEF vaccine virus was found to be as high as the current vaccines retaining the minimal titer required for YF 17D vaccines. The production passage in CEF did not lead to the selection of genetic variants as shown by nucleotide sequence analyses of the CEF-derived vaccine lots or the sequence of viruses recovered from monkeys experimentally inoculated with the CEF virus. YF 17DD virus produced in CEF was also indistinguishable from its seed lot virus parent in terms of plaque size and immunogenicity in mice and monkeys. Comparison of the CEF virus and the seed lot virus made in chicken embryo in the internationally accepted monkey neurovirulence test (MNVT) revealed a higher clinical score for the former. The differences in central nervous system (CNS) histological scores for monkeys inoculated with the chicken embryo and experimental CEF vaccines were at the borderline level of statistical significance. These data warrant further studies on the monkey attenuation of other batches of CEF-derived vaccines. PMID- 15752838 TI - Transcutaneous immunization with a 40-kDa outer membrane protein of Porphyromonas gingivalis induces specific antibodies which inhibit coaggregation by P. gingivalis. AB - This study seeks to assess the potential of a 40-kDa outer membrane protein of Porphyromonas gingivalis (40k-OMP) as a transcutaneous vaccine against chronic periodontitis. Transcutaneous immunization (TCI) of mice with 40k-OMP alone elicited 40k-OMP-specific IgG antibody (Ab) responses in both serum and saliva. When administered with cholera toxin (CT) as adjuvant, TCI with 40k-OMP not only elevated IgG Abs as noted above, but also induced IgA responses in serum but not in saliva. Salivary IgG from mice given 40k-OMP alone or 40k-OMP plus CT showed higher binding levels to the 40k-OMP than did that of non-immunized mice. Ab forming cell (AFC) analysis revealed high numbers of 40k-OMP-specific IgG AFCs in the spleen but low numbers in the salivary glands of mice given 40k-OMP alone or 40k-OMP plus CT. Since 40k-OMP-specific IgG inhibited the coaggregation of P. gingivalis vesicles and S. gordonii, TCI with 40k-OMP may be a useful tool in the quest to prevent P. gingivalis infection. PMID- 15752840 TI - Montanide ISA 720 vaccines: quality control of emulsions, stability of formulated antigens, and comparative immunogenicity of vaccine formulations. AB - Montanide ISA 720 is an experimental adjuvant, formulated as water-in-oil emulsions, that induces high antibody titers in several animal species. It has been used in human vaccine trials with malaria and HIV vaccines. The heightened response is likely due, in part, to the formation of a depot at the injection site. However, post-formulation modifications were seen with seven proteins tested during storage of ISA 720 formulations at 37 degrees C for 1 week and two proteins stored longer at 4 degrees C. Potency studies in mice, in which the stored vaccines were diluted into placebo emulsions for appropriate dosing, indicated that this instability could lead to loss of immunogenicity in the post injection depot, limiting the allowable storage time of preformed vaccines. We describe point-of-injection formulation for ISA 720 vaccines that meets the requirement for in vitro stability. For preformed vaccines, addition of glycine or glycylglycine prevented antigen modification on storage at 37 degrees C, providing a potential way of stabilizing antigen/ISA 720 formulations for in vitro storage and the post-injection depot. PMID- 15752839 TI - Antibody-dependent cell-mediated cytotoxic responses in participants enrolled in a phase I/II ALVAC-HIV/AIDSVAX B/E prime-boost HIV-1 vaccine trial in Thailand. AB - Antibody-dependent cell-mediated cytotoxicity (ADCC) was assessed in volunteers participating in an ALVAC-HIV (vCP1521)/AIDSVAX B/E gp120 prime-boost vaccine trial in Thailand. ADCC activity was measured using chromium release from gp120 subtype B- and CRF01_AE-coated targets in 95 vaccinees and 28 placebo recipients. There was a significant difference in the magnitude of the ADCC response to both targets between vaccinees and placebo recipients. The frequency of responders to subtype B and to CRF01_AE was 96% and 84% in the vaccine group versus 11% and 7% in the placebo group. The results demonstrate that this HIV vaccine is a potent inducer of ADCC activity and may be an additional protection of this prime-boost vaccine in preventing HIV disease. PMID- 15752841 TI - Antibodies generated by a novel DNA vaccination identify the MHC class III encoded BAT2 polypeptide. AB - Recombinant vaccines containing Ii sequences were employed to elicit an antibody response. Gene gun immunisation of mice with the recombinant Ii-antigen-encoding vectors induced antigen-specific antibodies. Antibody levels were substantially elevated when the DNA construct was extended by a sequence encoding the protease inhibitory domain of the invariant chain isoform Ii41. Employing this approach, we raised antibodies specific for a novel MHC class III encoded protein. The antibodies identify the 216 kDa BAT2 polypeptide. Immunostaining of embryonic tissue sections showed specific expression, especially in central nervous tissue, and suggests BAT2 as a novel differentiation marker. PMID- 15752843 TI - Solubility of chlorine in aqueous hydrochloric acid solutions. AB - The solubility of chlorine in aqueous hydrochloric acid solutions was studied. The effects of HCl concentration and temperature on the solubility were evaluated, and the thermodynamic parameters of the dissolution were calculated. It was found that the solubility isotherms had a minimum at about 0.5M HCl concentration at all the temperatures studied and that solubility decreased with the increase of temperature at all the HCl concentration range investigated. PMID- 15752842 TI - Some characteristics of 3,7-dinitro-, 3,7-dinitroso- and dinitrate compounds derived from 1,3,5,7-tetraazabicyclo[3.3.1]nonane. AB - The paper presents a set of some literature data and the authors' own experimental results of stability, sensitivity and explosion parameters of energetic Mannich N-bases, 3,7-dinitro-1,3,5,7-tetraazabicyclo[3.3.1]nonane (DPT), 3,7-dinitroso-1,3,5,7-tetraazabicyclo[3.3.1]nonane (DNPT) and hexamethylenetetramine dinitrate (HEXADI). Both their chemical and thermal reactivities are discussed. The results of small-scale cook-off test, determination of initiation ability, detonation velocity, impact sensitivity and performance show that the lowest process safety risks are connected with HEXADI. PMID- 15752844 TI - Thermal decomposition of methylene-4,4'-di(ethylphenyl-carbamate) to methylene 4,4'-di(phenylisocyanate). AB - During thermal decomposition of methylene-4,4'-di(ethylphenylcarbamate) (MDU), methylene-4,4'-di(phenylisocyanate) (MDI) appears to be the main product, provided that the ethanol formed in the reaction is carried away. Polycarbodiimides are the main by-products along with small amounts of 4,4' di(aminophenyl)methane and its derivatives. Under the reaction conditions used in these studies, the latter compound together with its derivatives, reacts both with MDI and MDU. At 220-310 degrees C and a reaction time of 80-110 min, the selectivity of the MDU transformation to MDI in relation to consumed MDU is 60-87 mol.%. The selectivity of the transformation to polycarbodiimides is 7-20 mol.%, and that to amines and urea derivatives is about 1-2 mol.%. PMID- 15752845 TI - Simple determination of performance of explosives without using any experimental data. AB - A simple procedure is introduced by which detonation pressure of CaHbNcOd explosives can be predicted from a, b, c, d and calculated gas phase heat of formation of explosives at any loading density without using any assumed detonation products and experimental data. It is shown here that the loading density, simply calculated heat of formation by additivity rule and atomic composition can be integrated into an empirical formula for predicting the detonation pressure of proposed explosives. Calculated detonation pressures by the introduced method for both pure and explosive formulations show good agreement with respect to measured detonation pressure over a wide range of loading density. The deviations are within about experimental errors. PMID- 15752846 TI - Effectiveness of expedient sheltering in place in a residence. AB - The objective of this study was to evaluate the effectiveness of expedient sheltering in place in a residence for protection against airborne hazards, as outlined in the U.S. Department of Homeland Security (DHS) guidance to the public. An improved method was developed to determine the air flow rate for a shelter inside a house. Expedient sheltering measures (plastic sheeting and duct tape) were applied to a room inside a test house by participants who followed the DHS guidance. Measured air flow rates were used to determine protection factors for various scenarios. Protection factors were calculated for the house and shelter under various occupancy times, weather conditions, and outdoor exposure times for hazardous agents. Protection factors ranged from 1.3 to 539, depending on the conditions. Results indicate that proper sealing can make a substantial difference in the effectiveness of the shelter. Sheltering in place can be most beneficial if people enter shelters before the arrival of a cloud of hazardous agent, and people exit shelters as soon as the cloud passes over. However, sheltering in place can be detrimental if people enter or exit shelters too late. CO2 and O2 concentrations inside the shelter are not likely to reach dangerous levels under most scenarios, but concentrations could reach dangerous levels under certain conditions, and concentration levels could affect individuals with respiratory problems. PMID- 15752847 TI - Accidental releases of titanium tetrachloride (TiCl4) in the context of major hazards--spill behaviour using REACTPOOL. AB - Titanium tetrachloride is a highly toxic and corrosive substance that is used widely in the process industries. On accidental release, it creates liquid pools that can either boil or evaporate. The main feature of the liquid pool is the reaction of titanium tetrachloride with water. There are three sources of water available for reaction: free ground water, atmospheric moisture and substrate water. Unfortunately, there is no specific study that examines the liquid phase hydrolysis reaction of titanium tetrachloride. Based on thermodynamic calculations and relevant information found on the topic, it is concluded that liquid titanium tetrachloride reacts exothermically with all three sources of water yielding hydrogen chloride gas and a solid complex of titanium oxychloride. The purpose of this paper is to describe the spill behaviour of titanium tetrachloride reporting a number of results using the REACTPOOL model [T. Kapias, R.F. Griffiths, C. Stefanidis, REACTPOOL: a code implementing a new multi compound pool model that accounts for chemical reactions and changing composition for spills of water reactive chemicals, J. Hazard. Mater. A81 (2001) 1-18]. It also addresses the dangers involved in cases of accidental release of titanium tetrachloride and reports its properties, referring to toxicity data and other relevant information. The spill behaviour of titanium tetrachloride has been incorporated into REACTPOOL. Model results indicate that the pool behaviour is mainly affected by the amount of free ground water, the wind speed and surface roughness. Although titanium tetrachloride has been involved in a number of major accidents, there are no experimental data relevant to the modelling requirements. PMID- 15752848 TI - Synthesis, characterization, thermolysis and performance evaluation studies on alkali metal salts of TABA and NTO. AB - The lithium (Li) and potassium (K) salts of 3-nitro-1,2,4-triazol-5-one (NTO) and 2,4,6-trinitroanilino benzoic acid (TABA) were prepared and characterized during this work. The synthesis was carried out by addition of a solution of lithium/potassium hydroxide to the aqueous solution of NTO and TABA, respectively. The products were characterized by elemental analysis, metal content determination and Fourier Transform Infrared (FTIR) Spectrum. Differential scanning calorimetry (DSC) profile indicated that Li and K salts of NTO and TABA undergo exothermic decomposition in the temperature range of 257-360 degrees C suggesting their energetic nature. The thermo gravimetric (TG) weight loss pattern revealed loss of water for Li/K salts of NTO and TABA in the temperature range of 115-155 degrees C. Sensitivity results revealed that the compounds are insensitive to impact and friction (impact sensitivity--height of 50% explosion>170 cm and friction insensitivity up to 36 kg) stimuli despite even the parent molecule of NTO salts (NTO) being HEM in the hazard category of 1.1. The FTIR spectra of the gaseous products evolved during TGA of NTO and TABA salts indicated the release of NO2. The formation of products such as LiNCO and KNCO was also observed in case of NTO salts, whereas that of CO2 and NH containing products was indicated in case of TABA salts during this study. In order to assess the performance as energetic ballistic modifiers (EBMs), NTO and TABA salts were incorporated in the ammonium perchlorate-hydroxyl terminated polybutadiene (AP-HTPB) composite propellants. The potassium salts enhanced the burning rate of the propellant. The best catalytic effect was obtained with K TABA salt, which increased the burning rate to the extent of approximately 81% as well as brought down the n-value to 0.15 (pressure 2-9 MPa). PMID- 15752849 TI - Spectro-thermal decomposition study of 1,4-dinitroglycoluril (DINGU). AB - Simultaneous thermal analysis and high temperature FTIR study of 1,4 dinitroglycoluril (DINGU) is reported. TG showed 90% weight loss in the temperature range 225-250 degrees C. Isothermal TG of DINGU showed about 70% weight loss in the temperature range 210-230 degrees C. Kinetic parameters evaluated using a computer program showed that alpha-t data are best described by the Avrami-Erofeev's equation for n=2 with an activation energy of 165 kJ/mol. The kinetics of decomposition of DINGU was followed by studying NH (3388 cm(-1)), CO (1770-1810 cm(-1)) and NO2 symmetric stretching (1565-1570 cm(-1)) IR bands. All three bands showed loss of intensity with temperature and time. alpha-t data of decomposition with respect to NO2 stretching was again best described by the Avrami-Erofeev's equation for n=2. Gaseous decomposition products observed in the IR were N2O, NO2, CO2, HCN and NO. PM3 and Hartree-Fock level calculations on various bond lengths, bond angles and dihedral angles were computed to support the analysis of decomposition study using TG and IR. The data showed that CN and NH bonds are much shorter than the NN bonds, indicative of the weaker NN bond and hence, the possibilities of rupture of the same bond preferentially. This paper also discusses the sensitivity and performance properties of DINGU. PMID- 15752850 TI - Environmental risk assessment of hydrofluoroethers (HFEs). AB - Hydrofluoroethers (HFEs) are being used as third generation replacements to chlorofluorocarbons (CFCs), hydrochlorofluorocarbons (HCFCs) and perfluorocarbons (PFCs) because of their nearly zero stratospheric ozone depletion and relatively low global warming potential. HFEs have been developed under commercial uses as cleaning solvents (incl., HFE-7500, C7F15OC2H5; HFE-7200, C4F9OC2H5; HFE-7100, C4F9CH3; HFE-7000, n-C3F7OCH3), blowing agents (incl., HFE-245mc, CF3CF2OCH3; HFC 356mec, CF3CHFCF2OCH3), refrigerants (incl., HFE-143a, CF3OCH3; HFE-134, CHF2OCHF2; HFE-245mc, CF3CF2OCH3), and dry etching agents in semiconductor manufacturing, (incl., HFE-227me, CF3OCHFCF3). From the environmental, ecological, and health points of view, it is important to understand their environmental risks for these HFEs from a diversity of commercial applications and industrial processes. This paper aims to introduce these HFEs with respect to physiochemical properties, commercial uses, and environmental hazards (i.e. global warming, photochemical potential, fire and explosion hazard, and environmental partition). Further, it addresses the updated data on the human toxicity, occupational exposure and potential health risk of commercial HFEs. It is concluded that there are few HFEs that still possess some environmental hazards, including global warming, flammability hazard and adverse effect of exposure. The partition coefficient for these HFEs has been estimated using the group contribution method; the values of logKow for commercial HFEs have been estimated to be below 3.5. PMID- 15752851 TI - Experimental study of flammability limits of natural gas-air mixture. AB - Flammability limits data are essential for a quantitative risk assessment of explosion hazard associated with the use of combustible gas. The present work is to obtain the fundamental flammability data for prevention of the hazards in the practical applications. Experiments have been conducted in a constant volume combustion bomb, and the fuel considered here is natural gas (NG). The pressure histories in the combustion bomb are recorded and a criterion of 7% pressure rise has been used to judge a flammable mixture. The effects of ethane on NG-air flammability limits have been investigated. By adding diluent (carbon dioxide, nitrogen or their mixture) into NG-air mixture, the dilution effects on the flammability limits have been explored as well, and the results are plotted as functions of diluent ratio. PMID- 15752852 TI - Comparison of fly ash properties from Afsin-Elbistan coal basin, Turkey. AB - Afsin-Elbistan (AE) coal fly ashes obtained by burning coal samples from top, middle and bottom sections of the AE coal seam were characterized and their properties were compared. Chemical analysis of the AE coal fly ashes showed that they are mainly composed of CaO, SiO2, Fe2O3 and Al2O3. Quantitative X-ray diffraction (XRD) analyses were carried out using an interactive data processing system (SIROQUANT) based on Rietveld interpretation methods. Lime is found in all the samples, ranging from around 7% to just over 38%. Amorphous contents of fly ashes are ranged between 19% and 25%. Different types of AE fly ashes revealed that bottom section coal fly ash is very similar to Class F, while medium and top section coal fly ashes are close to Class C and they might be used as mineral admixture in concrete. But also they do not comply with any of the standard. The results presented here show new possibilities for AE coal fly ashes in a wide range of fields, resulting in great advantages in waste minimization, as well as, resources conservation. PMID- 15752853 TI - Effect of salinity on nitrous oxide emission in the biological nitrogen removal process for industrial wastewater. AB - The effects of wastewater salinity on both nitrogen removal efficiency and N2O emission rate were investigated in a single nitrification process, a single denitrification process and an anoxic-oxic activated sludge process. In the single nitrification process, by increasing the salt concentration from 1.0 to 2.0 wt%, the N2O conversion ratio in the steady state increased by 2.2 times, from 0.22 to 0.48%. In the single denitrification process, a minimal change in the N2O conversion ratio was observed in the steady state even when the salt concentration was increased from 3.0 to 5.0 wt%. From the results of the anoxic oxic activated sludge process, it was found that a salt concentration increase from 1.6 to 3.0 wt% significantly increases the N2O conversion ratio from 0.7 to 13%. It is suggested that an increase in salt concentration markedly influences N2O emission both directly and indirectly via the inhibition of N2O reductase activity. The indirect inhibition is due to the high concentration of dissolved oxygen which is transported from the oxic tank to the anoxic tank through the circulated liquid. Thus, the salt concentration should be maintained below 3.0% to suppress N2O emission in an anoxic-oxic activated sludge process. PMID- 15752854 TI - Phenol and substituted phenols AOPs remediation. AB - The oxidation of phenol and two substituted species (4-nitrophenol and 4 chlorophenol) has been carried out by means of the O3, UV-vis, O3+UV-vis, TiO2+UV vis, O(3)+UV-vis+TiO2 and O3+TiO2 systems. From UV-vis experiments, the quantum yield of these organics has been calculated (0.018, 0.005 and 0.017 mol per Einstein for phenol, 4-nitrophenol and 4-chlorophenol, respectively). Broadly speaking, the addition of titania powder results in a slight inhibition of the parent compound degradation rate, although a positive effect is experienced when measuring the chemical oxygen demand (COD) and total organic carbon (TOC) removals. Amongst the technologies investigated, those combining ozone and radiation show the best efficiency in terms of phenols elimination and also COD and TOC decay rates. A simple economy analysis of the processes illustrates how the combinations O3+UV-vis and O3+UV-vis+TiO2 are the most attractive technologies, although some additional considerations have to be taken into account. PMID- 15752855 TI - Comparison of the treatment methods efficiency for decolorization and mineralization of Reactive Black 5 azo dye. AB - Degradation of Reactive Black 5 (RB5), a well-known non-biodegradable disazo dye, has been studied using UV/TiO2, wet-air oxidation (WAO), electro-Fenton (EF) and UV/electro-Fenton (UV/EF) advanced oxidation processes (AOPs). The efficiency of substrate decolorization and mineralization in each process has been comparatively discussed by decreases in concentration and total organic carbon content of RB5 solutions. The most efficient method on decolorization and mineralization was observed to be WAO process. Mineralization efficiency was observed in the order of WAO>UV/TiO2>UV/EF>EF. Final solutions of AOPs applications after 90 min treatment can be disposed safely to environment. Photocatalytic degradation kinetics of RB5 successfully fitted to Langmuir Hinshelwood (L-H) kinetics model. The values of second order degradation rate constant (k'') and adsorption constant (K) were determined as 5.085 mg L(-1)min( 1) and 0.112 L mg(-1), respectively. PMID- 15752856 TI - Sorption study of an acid dye from an aqueous solutions using modified clays. AB - The removal of the pollutant Supranol Yellow 4GL (S.Y.4GL) was studied by using different clays: clay exchanged with sodium (BNa+) and hydroxyaluminic polycation pillared clays in the presence or absence of non-ionic surfactant. While decomposing the surfactant at 500 degrees C, the surface of the clay changed significantly. The study of the behaviour of the three clays with respect to coloring solutions, allowed to determine the equilibrium time and the rate determining step of the dye S.Y.4GL adsorption. Two simplified kinetic models, were tested to investigate the adsorption mechanisms in terms of pseudo-first order and pseudo-second order equations. Besides, the adsorption capacity data were fitted to Langmuir and Freundlich equations as well. A better fixation was obtained with an acidic pH. The effect of temperature on the adsorption of dye has been also studied and the thermodynamic parameters DeltaG degrees , DeltaH degrees and DeltaS degrees were determined. PMID- 15752857 TI - Vitrified metal finishing wastes I. Composition, density and chemical durability. AB - Durable phosphate glasses were formed by vitrifying waste filter cakes from two metal finishing operations. Some melts formed crystalline components during cooling. Compositional analysis of dried, heat treated and vitrified samples was made using energy-dispersive X-ray spectroscopy, X-ray fluorescence spectroscopy, inductively-coupled plasma spectroscopy and Leco induction furnace combustion analysis. Hydrolytic dissolution, measured by an adapted product consistency test, was reduced by up to 3 orders of magnitude upon heat treatment or vitrification, surpassing the performance of borosilicate glass in some cases. This was attributed to the high levels of iron and zinc in the wastes, which greatly improve the durability of phosphate glasses. One of the wastes arose from a metal phosphating process and was particularly suitable for vitrification due to its high P2O5 content and favourable melting behaviour. The other waste, which arose from a number of processes, was less suitable as it had a low P2O5 content and during heating it emitted harmful corrosive gases and underwent violent reactions. Substantial volume reductions were obtained by heat treatment and vitrification of both wastes. Compositions and performances of some vitrified wastes were comparable with those of glasses which are under consideration for the immobilisation of toxic and nuclear wastes. PMID- 15752858 TI - Catalytic activity of active carbons impregnated before activation of pinewood sawdust and nutshells to be used on the control of atmospheric emissions. AB - This work analyses the catalytic activity of metal oxides impregnated on activated carbons to be used for the complete oxidation of benzene present in atmospheric emissions. When the impregnation step is performed before CO2 activation, the knowledge about catalytic activity is as yet quite scarce, being the main objective of the study here reported. Pinewood sawdust and nutshells were recycled to produce the activated carbons. Non-expensive metal oxides (CoO, Co3O4 and CrO3) were impregnated. When the impregnation was performed before CO2 activation instead of after activation, at 523 K the kinetic constants were 3.6 4.3 times higher for sawdust carbons, and 2.1-2.7 times higher for nutshell carbons, due to a better metal oxide dispersion on higher mesopore areas and on wider micropores. With Co3O4 as catalyst, a benzene conversion of 90% was reached at a lower temperature than with CrO3 (472 and 558 K, respectively). The carbons impregnated before CO2 activation allowed very good conversions at temperatures that guarantee carbon stability (lower than 575 K). The results obtained led to the conclusion that activated carbon is a suitable support for metal oxide catalyst aiming the complete oxidation of benzene, mainly if an adequately porous texture is induced, proceeding to the impregnation before CO2 activation. PMID- 15752859 TI - Characterization of the catalytic films formed on stainless steel anodes employed for the electrochemical treatment of cuprocyanide wastewaters. AB - Surface composition changes at stainless steel anodes in an electrochemical reactor applied for the electrochemical treatment of cuprocyanide-containing wastewaters operating under different hydrodynamic conditions were investigated. Under highly alkaline conditions in situ generation of a surface film on the anode with catalytic properties towards cyanide electrolysis was observed. X-ray photoelectron spectroscopy (XPS) results demonstrated that only copper oxi hydroxide compounds constitute the surface film developed on the stainless steel anodes, as no traces of N- and C-containing compounds were observed. The collected XPS spectra revealed relevant details concerning the oxidation states of copper in the film, and the products Cu2O, CuO and Cu(OH)2 were identified on the surface of the anodes. However, the quantitative proportions of the individual products differ and depend on the type of mixing employed during reactor operation. PMID- 15752860 TI - Effects of an electric or magnetic field on the radiolytic degradation of two biorefractory contaminants. AB - Effects of an electric or magnetic field on the radiolytic degradation of two biorefractory contaminants, Acid Orange 7 (AO7) and nitrobenzene (NB), were evaluated in this work. A continuous DC electric current with a low density (approximately 2.8-5.6 mA cm(-2)) applied during the radiolytic degradation of AO7 and NB solutions only led to slight enhancement in their degradation rate constants, but altered significantly the degradation mechanisms. On the other hand, application of a magnetic field (0.4 T) in irradiation processes slightly enhanced the degradation kinetics without leading to any change in degradation mechanisms. PMID- 15752861 TI - Sequencing batch reactor performance treating PAH contaminated lagoon sediments. AB - The applicability of sediment slurry sequencing batch reactors (SBR) to treat Venice lagoon sediments contaminated by polycyclic aromatic hydrocarbons (PAHs) was investigated, carrying out experimental tests. The slurry, obtained mixing tap water and contaminated sediments with 17.1 mg kg(-1) TS total PAHs content, was loaded to a 8l lab-scale completely stirred reactor, operated as a sequencing batch reactor. Oxygen uptake rate exerted by the slurry, measured by means of a DO-stat titrator, was used to monitor the in-reactor biological activity and to select the optimal operating conditions for the sediment slurry SBR. The PAHs removal efficiency was evaluated in different operating conditions, obtained changing the hydraulic retention time (HRT) of the lab-scale reactor and adding an external carbon source to the slurry. HRT values used during the experiments are 98, 70 and 35 days, whereas the carbon source was added in order to evaluate its effect on the biological activity. The results have shown a stable degradation of PAHs, with a removal efficiency close to 55%, not dependent on the addition of carbon source and the tested HRTs. PMID- 15752862 TI - Assessing effect of electrode configuration on the efficiency of electrokinetic remediation by sequential extraction analysis. AB - The electroremediation experiments were conducted on artificially polluted soils by introducing a single metallic contaminant (Pb, Zn and Cu) and multiple metallic contaminants (Pb+Zn+Cu). Based on sequential extraction results, it was observed that the removal efficiencies of lead, zinc and copper vary depending on types of contamination. When the soil was contaminated only by lead, the removal efficiency was found to be 48%. However, the removal efficiency of lead decreased to 32% when the soil was contaminated by the combination of lead, zinc and copper. Similar results were observed for zinc and copper. The corresponding removal efficiency values for zinc and copper were 92% and 37%, and 34% and 31%, respectively. Effects of electrode geometry on the removal efficiency of metals were investigated by constructing a multiple anode arrangement. In this arrangement, the electrokinetic unit consists of three cylinders, which lie one inside the other, and the soil was placed in the middle cylinder. The central cylinder was the cathode well and the outer cylinder was the anode well, where eight identical anode electrodes were placed in octagonal with respect to the cathode electrode. By using this electrode arrangement in removal of metals from the soil contaminated with the combination of three metals (Pb+Zn+Cu), the removal efficiencies of lead, zinc and copper were found to be 29%, 18% and 18%, respectively. As it can be seen, these numerical values are much lower than the values that were obtained when the traditional two-plate electrode arrangement used in the electroremediation experiments (32%, 37% and 31%). PMID- 15752863 TI - Removal of Cr(VI) from aqueous solution by two Lewatit-anion exchange resins. AB - The sorption of hexavalent chromium, Cr(VI), from aqueous solutions on macroporous resins containing tertiary amine groups (Lewatit MP 62 and Lewatit M 610) was studied at varying Cr(VI) concentrations, adsorbent dose, pH, contact time and temperatures. The concentration of chromium in aqueous solution was determined by inductively coupled plasma spectrometry (ICP-AES). Batch shaking sorption experiments were carried out to evaluate the performance of Lewatit MP 62 and Lewatit M 610 anion exchange resins in the removal of Cr(VI) from aqueous solutions. The ion-exchange process, which is pH dependent, shows maximum removal of Cr(VI) in the pH range 2-6 for an initial Cr(VI) concentration of 100 ppm. The sorption increases with the decrease in pH and slightly decreases with the increase in temperature. Both ion exchangers had high bonding constants with Lewatit M 610 showing stronger binding. It was observed that the maximum adsorption capacity of 0.40 mmol of Cr(VI)/g for Lewatit MP 62 and 0.41 mmol of Cr(VI)/g for Lewatit M 610 was achieved at pH of 5.0. The thermodynamic parameters (free energy change, DeltaG degrees ; enthalpy change, DeltaS degrees ; and entropy change, DeltaH degrees ) for the sorption have been evaluated. The rise in temperature caused a slight decrease in the value of the equilibrium constant (Kc) for the sorption of Cr(VI) ion. The sorption of Cr(VI) on the resin was rapid during the first 15 min and equilibrium was found to be attained within 30 min. The sorption of Cr(VI) onto the resins followed reversible first-order rate kinetics. Such ion exchange resins can be used for the efficient removal of chromium from water and wastewater. PMID- 15752864 TI - Removal of lead from aqueous solutions by activated phosphate. AB - The potential of using activated phosphate as a new adsorbent for the removal of Pb from aqueous solutions was investigated. The kinetic of lead adsorption and the adsorption process were compared for natural phosphate (NP) and activated phosphate (AP). The results indicate that equilibrium was established in about 1h for NP and 3 h for AP. The effect of the pH was examined in the range 2-6. The maximum removal obtained is between two and three for NP and between three and four for AP. The maximum adsorption capacities at 25 degrees C are 155.04 and 115.34 mg/g for AP and NP, respectively. The effect of temperature has been carried out at 25, 35 and 45 degrees C. The data obtained from adsorption isotherms of lead at different temperatures fit to linear form of Langmuir adsorption equation. The thermodynamic parameters such as enthalpy (DeltaH), free energy (DeltaG) and entropy (DeltaS) were calculated. They show that adsorption of lead on NP and AP is an endothermic process more effective at high temperatures. These results show that AP is a good adsorbent for heavy metals from aqueous solutions and could be used as a purifier for water and wastewater. PMID- 15752865 TI - Determination of the adsorption capacity of activated carbon made from coffee grounds by chemical activation with ZnCl2 and H3PO4. AB - In order to evaluate the adsorptive capacities of granular activated carbon produced from coffee grounds by chemical activation, the adsorption of different phenols and acid and basic dyes, has been carried out. The comparison with a commercial activated carbon has been made. Adsorption isotherms of phenols and dyes (acid and basic) onto produced and commercial granular activated carbons were experimentally determined by batch tests. Both Freundlich and Langmuir models are well suited to fit the adsorption isotherm data. As a result, the coffee grounds based activated carbon may be promising for phenol and dye removal from aqueous streams. PMID- 15752866 TI - The effect of ionic strength and hardness of water on the non-ionic surfactant enhanced remediation of perchloroethylene contamination. AB - The objective of this study is to evaluate the perchloroethylene (PCE) removal by an aqueous surfactant solutions based on influential factors (ionic strength, hardness) of various groundwaters and surface waters contaminated with PCE. The experimental methods used in this study were separatory funnel experiments and batch experiments. Separatory funnel experiments were performed to determine which surfactants are good solubilizers for PCE. Batch experiments were performed to evaluate the effect of ions in sampled water for PCE removal. The results of separatory funnel experiments indicated that the surfactant polyoxyethylene (20) sorbitan monostearate (Tween 60) showed to be a predominant solubilizer for the removal of PCE (87.3%). Separatory funnel experiments also showed that the hydrophilic-lipophilic balance (HLB) number and the chemical structure of the surfactants were good indicators of surfactant effectiveness for removal of PCE from water. The results of batch experiments showed that non-ionic surfactants are affected by the ionic strength of sampled water. The % of PCE removal of the Tween 60 surfactant solution was measured to be 88.3% by batch experiments. This result was affected by the characteristics of the surfactant (HLB, chemical structures) and the ionic strength of water. Therefore, the ionic strength of contaminated water, HLB and chemical structures of surfactants must be considered in surfactant-enhanced remediation. PMID- 15752867 TI - A novel solubilization of phenanthrene using Winsor I microemulsion-based sodium castor oil sulfate. AB - Problems associated with polycyclic aromatic hydrocarbons (PAHs) contaminated site in environmental media have received increasing attention. Ex situ soil washing is commonly used for treating contaminated soils by separating the most contaminated fraction of the soil for disposal. Surfactant-enhanced soil washing is being considered with increasing frequency to actually achieve soil contaminant separation. In this research, a novel solubilization of phenanthrene and extraction of phenanthrene from spiked soil by sodium castor oil sulfate (SCOS) microemulsion was presented and compared with the conventional surfactants, Triton X-100 (TX100), Tween 80 (TW80), Brij35, sodium dodecylbenzene sulfonate (SDBS) and sodium dodecyl sulfate (SDS). Unlike conventional surfactants, SCOS forms stable microemulsion in water and thus behaves much like a separate bulk phase in concentrating organic solutes. The extent of solubility enhancement is linearly proportional to the concentration of SCOS microemulsion, in contrast with the effect of a conventional surfactant in which a sharp inflection occurs in the vicinity of the measured critical micelle concentration. SCOS microemulsion exhibits the largest mass solubilization ratio among the selected surface active agents (SAAs) in both soil-free system and soil-water system. The partitioning coefficients of phenanthrene between the emulsified phase and the aqueous phase, Kem, is slightly larger than those between the micellar pseudo phase and the aqueous phase, Kmc. The extraction experiments demonstrate high and fast desorption of phenanthrene from spiked soil by SCOS microemulsion perhaps due to its high solubilization capacity compared with the conventional surfactant solutions. The results show that SCOS could be an attractive alternative to synthetic surfactants in ex situ washing for PAH contaminated soils. PMID- 15752868 TI - Analysis of arsenic and some other elements in coal fly ash by X-ray photoelectron spectroscopy. AB - Surface characterization of coal fly ash (CFA) was carried out by use of X-ray photoelectron spectroscopy (XPS), especially focusing on the occurrence of As. A peak in the XPS spectrum of CFA was assigned to oxide forms of As(3d). The molar ratios of Al, As, Ca, Fe, and S normalized to Si were obtained from XPS analysis (MR-X). Also, the molar ratios of those elements were calculated from bulk analysis (total element concentration in CFA) (MR-B). The MR-X/MR-B ratio of As was much higher than those of other elements, suggesting that As is highly enriched on the surface of CFA. When eight CFA samples were analyzed, there was an approximate relationship between the MR-X values and MR-B values for As. The leaching of elements from CFA was examined by XPS analysis and by bulk analysis. The leaching tests using EDTA and HNO3 resulted in a great decrease in the As(3d) peak area; the %leaching of As obtained by XPS analysis was almost equal to that by bulk analysis. PMID- 15752869 TI - Modification of surface properties of Lentinus sajor-caju mycelia by physical and chemical methods: evaluation of their Cr6+ removal efficiencies from aqueous medium. AB - The hexavalent chromium biosorption onto untreated and heat-, acid- and alkali treated Lentinus sajor-caju mycelia were studied from aqueous solutions. The particles sizes of the fungal mycelia ranged from 100 to 200 microm. The effect of pH, temperature, biosorbent dose, initial concentration of chromium ions, contact time parameters were investigated in a batch system. Biosorption equilibrium was established in about 4 h. The surface charge density of the fungal preparations varied with pH, and the maximum absorption of chromium ions on the fungal preparations were obtained at pH 2.0. The biosorption of chromium ions by the tested fungal preparations increased as the initial concentration of chromium ions increased in the medium. The maximum biosorption capacities of the untreated and heat, HCl- and NaOH-treated fungal biomass were 0.363, 0.613, 0.478 and 0.513 mmol Cr6+ per gram of dry biomass, respectively. The correlation regression coefficients and the Langmuir constant values show that the biosorption process can be well defined by Langmuir equation. The chromium adsorption data were analysed using the first- and the second-order kinetic models. The first-order equation is the most appropriate equation to predict the biosorption capacities of all the fungal preparations. In addition, the polarity and surface energy of the untreated and all the modified biomass film preparations were determined by contact angle measurement. All the tested fungal biomass preparations could be regenerated using 0.1 M NaOH solution. PMID- 15752870 TI - Adsorption of mercury by carbonaceous adsorbents prepared from rubber of tyre wastes. AB - Rubber from tyre wastes has been used to prepare carbonaceous adsorbents and the products obtained have been tested as adsorbents for mercury in aqueous solution. The adsorbents have been prepared by applying thermal, chemical and combined (thermal and chemical or vice versa) treatments. Tyre rubber has been: heated at 400 or 900 degrees C for 2 h in N2, chemically-treated with H2SO4, HNO3 or H2SO4/HNO3 solution for 24 h, and in two successive steps first heated at 400 degrees C for 2h in N2 and then treated with a H2SO4/HNO3 solution for 24 h, or vice versa. Resultant products have been characterised in terms of elementary composition and textural properties. The adsorption of mercury has been studied from kinetic and equilibrium standpoints. The treatments effected to tyre rubber decrease the carbon content and the hydrogen content. The oxygen content and the nitrogen content increase for the chemically-treated products. The heat treatment of tyre rubber results in a larger development of surface area, microporosity, and mesoporosity than the chemical treatments. These treatments, however, produce a great creation of macropores. In comparison to the starting rubber, the adsorption process of mercury is faster when the material is heated or treated with the H2SO4, HNO3 or 1:3 H2SO4/HNO3 solution. These adsorbents are either a non-porous solid or possess a high mesopore volume or a wide pore size distribution in the macropore range. The adsorption capacity is larger for products prepared by heat, chemical and combined treatments of the rubber. A common textural characteristic of these adsorbents is their better developed microporosity. The ability to adsorb mercury is higher for the heated products than for the chemically-treated ones. The maximum adsorption of mercury is 211 mg g(-1). The constant Kf of the Freundlich equation is as high as 108.9 mg g(-1). PMID- 15752871 TI - Effect of easily degradable substrate on anaerobic degradation of pentachlorophenol in an upflow anaerobic sludge blanket (UASB) reactor. AB - The effect of microbial easily degradable substrate (MEDS) on the anaerobic degradation of pentachlorophenol (PCP) in two upflow anaerobic sludge blanket (UASB) reactors was investigated. The results indicated that glucose-utilizing activity decreased with the increase of PCP concentration in the mixed culture, and MEDS promoted PCP-dechlorination and degrading activities. The concentration of MEDS increased from 917 to 4580 mg L(-1) with the increase of PCP concentration from 100 to 181 mg L(-1)in influent, which was necessary for maintenance of steady operation of the experimental reactors, the removal rate of PCP and COD ranged up to 99.5 and 90.0% and the concentration of PCP in the effluent was less than 0.5 mg L(-1). The concentration of PCP in effluent was linearly or logarithmically related to sucrose concentration in the influent while PCP was less than the maximum permissible PCP concentration. The activity of anaerobic sludge in the reactor decreased as the concentration of PCP increased, but it could be recovered step by step as time progressed. In the lowest layer of the reactor, the activity of sludge was the highest. So it is feasible to accelerate the degradation of the organic toxic compounds like PCP, by adding suitable quantities of microbial easily degradable substrate to the system. PMID- 15752872 TI - Total concentrations and fractions of Cd, Cr, Pb, Cu, Ni and Zn in sewage sludge from municipal and industrial wastewater treatment plants. AB - Heavy metals are one of the important factors that affect the final disposal of sewage sludge. In this paper, the metal mobility and bioavailability of heavy metals in sewage sludge were studied by using Community Bureau of Reference (BCR) sequential extraction procedure to get more information for the reasonable disposal of sludge. Sewage sludge was collected from five municipal wastewater treatment plants and three industrial wastewater treatment plants. The sludge was examined for and the total concentrations and different chemical fractions of Cd, Cr, Pb, Cu, Ni and Zn. The total metal concentrations of heavy metals in sludge varied greatly. The contents of Zn and Cu were the highest, followed by then Cr, Ni and Pb and the content of Cd was the least. There was no significant difference in total metal concentration between municipal and industrial wastewater treatment plants. Fractions extracted by the BCR sequential procedure were acid soluble/exchangeable, reducible and oxidizable fraction. Sludge pH was found to have profound effect on the chemical fractions of heavy metals. Acidic sludges (Xiamen and Jinlin Petrochemical Group Co., wastewater treatment plant) had higher proportion of the acid soluble/exchangeable fractions than in neutral sludge. In neutral sludges, Pb and Cr were principally distributed in between the oxidizable fraction and the residual fraction; Cu was in the oxidizable fraction; Cd mainly in the residual fraction in municipal wastewater treatment plants and had high percentage of acid soluble/exchangeable and reducible fractions in industrial wastewater treatment plants; Ni and Zn had higher percentage in the acid soluble/exchangeable and the oxidizable fraction. PMID- 15752873 TI - Crab shell-based biosorption technology for the treatment of nickel-bearing electroplating industrial effluents. AB - This paper discusses the possible application of a biosorption system with acid washed crab shells in a packed bed up-flow column for the removal of nickel from electroplating industrial effluents. Between two nickel-bearing effluents, effluent-1 is characterized by considerable amount of light metals along with trace amounts of lead and copper. Effluent-2 is characterized by relatively low conductivity, total dissolved solids and total hardness compared to effluent-1. Crab shells exhibited uptakes of 15.08 and 20.04 mg Ni/g from effluent-1 and effluent-2, respectively. The crab shell bed was regenerated using 0.01 M EDTA (pH 9.8, aq. NH3) and reused for seven sorption-desorption cycles. The EDTA elution provided elution efficiencies up to 99% in all the seven cycles. This, together with the data from regeneration efficiencies for seven cycles, provided evidence that the reusability of crab shell in the treatment of nickel-bearing electroplating industrial effluents is viable. PMID- 15752874 TI - A challenge to the therapeutic nihilism of ESPAC-1. PMID- 15752875 TI - The multidisciplinary clinic. PMID- 15752876 TI - Application of the No Action Level (NAL) protocol to correct for prostate motion based on electronic portal imaging of implanted markers. AB - PURPOSE: To evaluate the efficacy of the No Action Level (NAL) off-line correction protocol in the reduction of systematic prostate displacements as determined from electronic portal images (EPI) using implanted markers. METHODS AND MATERIALS: Four platinum markers, two near the apex and two near the base of the prostate, were implanted for localization purposes in patients who received fractionated high dose rate brachytherapy. During the following course of 25 fractions of external beam radiotherapy, the position of each marker relative to the corresponding position in digitally reconstructed radiographs (DRRs) was measured in EPI in 15 patients for on average 17 fractions per patient. These marker positions yield the composite displacements due to both setup error and internal prostate motion, relative to the planning computed tomography scan. As the NAL protocol is highly effective in reducing systematic errors (recurring each fraction) due to setup inaccuracy alone, we investigated its efficacy in reducing systematic composite displacements. The analysis was performed for the center of mass (COM) of the four markers, as well as for the cranial and caudal markers separately. Furthermore, the impact of prostate rotation on the achieved positioning accuracy was determined. RESULTS: In case of no setup corrections, the standard deviations of the systematic composite displacements of the COM were 3-4 mm in the craniocaudal and anterior-posterior directions, and 2 mm in the left-right direction. The corresponding SDs of the random displacements (interfraction fluctuations) were 2-3 mm in each direction. When applying a NAL protocol based on three initial treatment fractions, the SDs of the systematic COM displacements were reduced to 1-2 mm. Displacements at the cranial end of the prostate were slightly larger than at the caudal end, and quantitative analysis showed this originates from left-right axis rotations about the prostate apex. Further analysis revealed that significant time trends are present in these prostate rotations. No significant trends were observed for the prostate translations. CONCLUSIONS: The NAL protocol based on marker positions in EPI halved the composite systematic displacements using only three imaged fractions per patient, and thus allowed for a significant reduction of planning margins. Although large rotations of the prostate, and time trends therein, were observed, the net impact on the measured displacements and on the accuracy obtained with NAL was small. PMID- 15752877 TI - Prostate motion and isocenter adjustment from ultrasound-based localization during delivery of radiation therapy. AB - PURPOSE: To study prostate motion from 4,154 ultrasound alignment fractions on 130 prostate patients treated with conformal radiotherapy or intensity-modulated radiation therapy at the University of Nebraska Medical Center. METHODS AND MATERIALS: Each prostate patient was immobilized in a vacuum cradle. Daily treatment was verified by ultrasound scan after laser setup with skin marks and before radiation delivery by the same physician responsible for anatomic delineation during planning. Directional statistics were employed to test the significance of shift directions. RESULTS: Polar histograms showed the prevalence of prostate motion in superior-posterior directions. The average direction was about 27 degrees from the superior axis. The average changes of prostate position in superior to inferior (SI), anterior-posterior (AP), and left to right (LR) directions and in radial distance were 0.25, -0.13, 0.03, and 0.92, cm respectively. Our data indicated that prostate motion was not patient specific, and its average magnitude remained virtually unchanged over time. Recommended planning target volume (PTV) margins for use without ultrasound localization were 0.90 cm in SI, 1.02 cm in AP, and 0.80 cm in LR directions. CONCLUSION: Ultrasound localization revealed a predominance of prostate shift from planning position in the superior-posterior direction, with an average closer to the superior axis. The motion data provides recommended margins for PTV. PMID- 15752878 TI - Dose-response characteristics of low- and intermediate-risk prostate cancer treated with external beam radiotherapy. AB - PURPOSE: In this era of dose escalation, the benefit of higher radiation doses for low-risk prostate cancer remains controversial. For intermediate-risk patients, the data suggest a benefit from higher doses. However, the quantitative characterization of the benefit for these patients is scarce. We investigated the radiation dose-response relation of tumor control probability in low-risk and intermediate-risk prostate cancer patients treated with radiotherapy alone. We also investigated the differences in the dose-response characteristics using the American Society for Therapeutic Radiology and Oncology (ASTRO) definition vs. an alternative biochemical failure definition. METHODS AND MATERIALS: This study included 235 low-risk and 387 intermediate-risk prostate cancer patients treated with external beam radiotherapy without hormonal treatment between 1987 and 1998. The low-risk patients had 1992 American Joint Committee on Cancer Stage T2a or less disease as determined by digital rectal examination, prostate-specific antigen (PSA) levels of < or =10 ng/mL, and biopsy Gleason scores of < or =6. The intermediate-risk patients had one or more of the following: Stage T2b-c, PSA level of < or =20 ng/mL but >10 ng/mL, and/or Gleason score of 7, without any of the following high-risk features: Stage T3 or greater, PSA >20 ng/mL, or Gleason score > or =8. The logistic models were fitted to the data at varying points after treatment, and the dose-response parameters were estimated. We used two biochemical failure definitions. The ASTRO PSA failure was defined as three consecutive PSA rises, with the time to failure backdated to the mid-point between the nadir and the first rise. The second biochemical failure definition used was a PSA rise of > or =2 ng/mL above the current PSA nadir (CN + 2). The failure date was defined as the time at which the event occurred. Local, nodal, and distant relapses and the use of salvage hormonal therapy were also failures. RESULTS: On the basis of the ASTRO definition, at 5 years after radiotherapy, the dose required for 50% tumor control (TCD(50)) for low-risk patients was 57.3 Gy (95% confidence interval [CI], 47.6-67.0). The gamma50 was 1.4 (95% CI, -0.1 to 2.9) around 57 Gy. A statistically significant dose-response relation was found using the ASTRO definition. However, no dose-response relation was noted using the CN + 2 definition for these low-risk patients. For the intermediate-risk patients, using the ASTRO definition, the TCD(50) was 67.5 Gy (95% CI, 65.5-69.5) Gy and the gamma50 was 2.2 (95% CI, 1.1-3.2) around TCD(50). Using the CN + 2 definition, the TCD(50) was 57.8 Gy (95% CI, 49.8-65.9) and the gamma50 was 1.4 (95% CI, 0.2-2.5). Recursive partitioning analysis identified two subgroups within the low-risk group, as well as the intermediate-risk group: PSA level <7.5 vs. > or =7.5 ng/mL. Most of the benefit from the higher doses for the low- and intermediate-risk group was derived from the patients with the higher PSA values. For the low-risk group, the dose-response curves essentially plateaued at 78 Gy. CONCLUSION: A dose-response relation was found using the ASTRO definition for low risk prostate cancer. However, we found only marginal or no dose-response relation when the CN + 2 definition was used. Most of the benefit from the higher doses derived from low-risk patients with higher PSA levels. In all cases, little projected gain appears to exist at doses >78 Gy for these patients. A dose response relation was noted for the intermediate-risk patients using either the CN + 2 or ASTRO definition. Most of the benefit from the higher doses also derived from the intermediate-risk patients with higher PSA levels. Some room for improvement appears to exist with additional dose increases in this group. PMID- 15752879 TI - What pretreatment prostate-specific antigen level warrants long-term androgen deprivation? AB - PURPOSE: Several large randomized prospective studies have demonstrated a survival benefit with the addition of long-term androgen deprivation to definitive radiotherapy for patients with Gleason score 8-10 or T3-T4 prostate cancer. However, these studies were performed before the routine use of prostate specific antigen (PSA) measurement. The purpose of this study was to determine what pretreatment (initial) PSA (iPSA) level, if any, warrants the addition of long-term androgen deprivation in the PSA era. METHODS AND MATERIALS: The data set evaluated consisted of 1003 prostate cancer patients treated definitively with three-dimensional conformal radiotherapy between May 1, 1989 and November 30, 1999 (median follow-up, 61 months). Specifically excluded were patients with T3-T4 disease or Gleason score greater than 7 or those who had undergone androgen deprivation as a part of their initial therapy. The median radiation dose was 76 Gy. Patients were randomly split into two data sets, with the first (n = 487) used to evaluate the optimal iPSA cutpoint for which a statistically significant difference in outcome was noted. The second data set (n = 516) served as a validation data set for the initial modeling. The analysis of the optimal iPSA cutpoint was based on a recursive partitioning approach for censored data using the log-rank test for nodal separation of freedom from biochemical failure (FFBF) as defined by the American Society for Therapeutic Radiology and Oncology definition. Cox multivariate regression analysis was used to confirm independent predictors of outcome among the clinical and treatment-related factors: iPSA (grouped as defined by the recursive partitioning analysis), Gleason score (2-6 vs. 7), T stage (T1c-T2a vs. T2b-T2c), and total radiation dose (continuous). RESULTS: The recursive partitioning analysis data set resulted in an optimal iPSA cutpoint of 35 ng/mL, such that the 5-year Kaplan-Meier estimate of FFBF was 80%, 69%, and 19% for iPSA groups of 0-9.9, 10-35, and >35 ng/mL, respectively. The validation data set demonstrated the optimal iPSA cutpoint to be 30 ng/mL. Conservatively choosing 30 ng/mL as the optimal cutpoint, the 5-year FFBF estimate for the entire 1003 patients was 82%, 69%, and 20% for iPSA groups 0-9.9 (n = 630), 10-30 (n = 329), and >30 (n = 44) ng/mL, respectively. On multivariate regression analysis, with the iPSA grouped as above, the Gleason score and radiation dose were independent predictors of outcome in this patient group (all p < 0.001). On univariate analysis, a higher radiation dose improved FFBF when the iPSA level was between 10 and 30 ng/mL (p = 0.001) but not when the iPSA level was >30 or <10 ng/mL. CONCLUSION: Recursive partitioning techniques defined an iPSA cutpoint of 30 ng/mL for delineating intermediate vs. high risk. Patients with a PSA level >30 ng/mL in the absence of Gleason score >7 or T3 disease do poorly when treated with radiotherapy alone and should be considered for long term androgen deprivation or other aggressive systemic therapy. PMID- 15752880 TI - Gastrointestinal toxicity and its relation to dose distributions in the anorectal region of prostate cancer patients treated with radiotherapy. AB - PURPOSE: To study the correlations between the dose distributions in the anorectal region and late GI symptoms in patients treated for localized prostate carcinoma. METHODS AND MATERIALS: Data from a randomized study were analyzed. In this trial, patients were treated with either rectangular or conformal fields with a dose of 66 Gy. Data concerning GI symptoms were collected from questionnaires of 197 patients. The distributions of the anorectal region were projected on maps, and the dose parameters were calculated. The incidences of complaints were studied as a function of the dose-area parameters and clinical parameters, using a proportional hazard regression model. Finally, we tested a series of dose parameters originating from different parts of the anorectal region. RESULTS: Analyzing the total region, only a statistically significant dose-area effect relation for bleeding was found (p < 0.01). Defining subareas, we found effect relations for bleeding, soiling, fecal incontinence, and mucus loss. For bleeding and mucus loss, the strongest correlation was found for the dose received by the upper 70-80% of the anorectal region (p < 0.01). For soiling and fecal incontinence, we found the strongest association with the dose to the lower 40-50% (p < 0.05). CONCLUSION: We found evidence that complaints originate from specific regions of the irradiated lower GI tract. Bleeding and mucus loss are probably related to irradiation of the upper part of the rectum. Soiling and fecal incontinence are more likely related to the dose to the anal canal and the lower part of the rectum. PMID- 15752881 TI - Acute and late complications after radiotherapy for prostate cancer: results of a multicenter randomized trial comparing 68 Gy to 78 Gy. AB - PURPOSE: To compare acute and late gastrointestinal (GI) and genitourinary (GU) side effects in prostate cancer patients randomized to receive 68 Gy or 78 Gy. METHODS AND MATERIALS: Between June 1997 and February 2003, 669 prostate cancer patients were randomized between radiotherapy with a dose of 68 Gy and 78 Gy, in 2 Gy per fraction and using three-dimensional conformal radiotherapy. All T stages with prostate-specific antigen (PSA) <60 ng/mL were included, except any T1a and well-differentiated T1b-c tumors with PSA < or =4 ng/mL. Stratification was done for four dose-volume groups (according to the risk of seminal vesicles [SV] involvement), age, hormonal treatment (HT), and hospital. The clinical target volume (CTV) consisted of the prostate with or without the SV, depending on the estimated risk of SV invasion. The CTV-planning target volume (PTV) margin was 1 cm for the first 68 Gy and was reduced to 0.5 cm (0 cm toward the rectum) for the last 10 Gy in the 78 Gy arm. Four Dutch hospitals participated in this Phase III trial. Evaluation of acute and late toxicity was based on 658 and 643 patients, respectively. For acute toxicity (<120 days), the Radiation Therapy Oncology Group (RTOG) scoring system was used and the maximum score was reported. Late toxicity (>120 days) was scored according to the slightly adapted RTOG/European Organization for Research and Treatment of Cancer (EORTC) criteria. RESULTS: The median follow-up time was 31 months. For acute toxicity no significant differences were seen between the two randomization arms. GI toxicity Grade 2 and 3 was reported as the maximum acute toxicity in 44% and 5% of the patients, respectively. For acute GU toxicity, these figures were 41% and 13%. No significant differences between both randomization arms were seen for late GI and GU toxicity, except for rectal bleeding requiring laser treatment or transfusion (p = 0.007) and nocturia (p = 0.05). The 3-year cumulative risk of late RTOG/EORTC GI toxicity grade > or =2 was 23.2% for 68 Gy, and 26.5% for 78 Gy (p = 0.3). The 3-year risks of late RTOG/EORTC GU toxicity grade > or =2 were 28.5% and 30.2% for 68 Gy and 78 Gy, respectively (p = 0.3). Factors related to acute GI toxicity were HT (p < 0.001), a higher dose-volume group (p = 0.01), and pretreatment GI symptoms (p = 0.04). For acute GU toxicity, prognostic factors were: pretreatment GU symptoms (p < 0.001), HT (p = 0.003), and prior transurethral resection of the prostate (TURP) (p = 0.02). A history of abdominal surgery (p < 0.001) and pretreatment GI symptoms (p = 0.001) were associated with a higher incidence of late GI grade > or =2 toxicity, whereas HT (p < 0.001), pretreatment GU symptoms (p < 0.001), and prior TURP (p = 0.006) were prognostic factors for late GU grade > or =2. CONCLUSIONS: Raising the dose to the prostate from 68 Gy to 78 Gy resulted in higher incidences of acute and late GI and GU toxicity, but these differences were not significant, except for late rectal bleeding requiring treatment and late nocturia. Other factors than the studied dose levels appeared to be important in predicting toxicity after radiotherapy, especially previous surgical interventions (abdominal surgery or TURP), hormonal therapy, and the presence of pretreatment symptoms. PMID- 15752882 TI - Toward a definition of a threshold for harmless doses to the anal-sphincter region and the rectum. AB - PURPOSE: To investigate dysfunction caused by unwanted radiation to the anal sphincter region and the rectum. METHODS AND MATERIALS: A questionnaire assessing bowel symptoms, sexual function, and urinary symptoms was sent to 72 patients with clinically localized prostatic adenocarcinoma treated by external beam radiation therapy at the Radiumhemmet, Karolinska Hospital, in Stockholm, Sweden, 2-4 years after treatment. The mean percentage dose-volume histograms for patients with and without the specific symptom were calculated. RESULTS: Of the 65 patients providing information, 9 reported fecal leakage, 10 blood and mucus in stools, 10 defecation urgency, and 7 diarrhea or loose stools. None of the 19 and 13 patients who received, respectively, a dose of > or =35 Gy to < or =60% or > or =40 Gy to < or =40% of the anal-sphincter region volume reported fecal leakage (p < 0.05). In dose-volume histograms, a statistically significant correlation was found between radiation to the anal-sphincter region and the risk of fecal leakage in the interval 45-55 Gy. There was also a statistically significant correlation between radiation to the rectum and the risk of defecation urgency and diarrhea or loose stools in the interval 25-42 Gy. No relationship was found between anatomic rectal wall volume and the investigated late effects. CONCLUSIONS: Although the limited data in this study prevent the definition of a conclusive threshold regarding volume and dose to the anal sphincter region and untoward morbidity, it seems that careful monitoring of unnecessary irradiation to this area should be done because it can potentially help reduce the risk of adverse effects, such as fecal leakage. Future studies should pay more attention to the anal-sphincter region and help to more rigorously define its radiotherapeutic tolerance. PMID- 15752883 TI - Concomitant radiation therapy and paclitaxel for unresectable locally advanced breast cancer: results from two consecutive phase I/II trials. AB - PURPOSE: The management of unresectable locally advanced breast cancer (ULABC) remains a major challenge because of the necessity both to treat local disease and to prevent distant disease. Two consecutive Phase I/II trials of concomitant chemotherapy and radiation (CRT) were performed to attempt to address both local and distant disease control in ULABC. This analysis focuses on rates of locoregional control and radiation-associated acute and late complications. METHODS AND MATERIALS: Thirty-three patients with unresectable locally advanced or inflammatory breast cancers (T4N0-3M0-1) or locally recurrent disease were treated with CRT on two consecutive Phase I/II trials. Radiotherapy consisted of 60-70 Gy to the breast or chest wall and 60 Gy to draining lymphatics in a week on/week-off (WO/WO) schedule. Chemotherapy consisted of either continuous infusion or bolus paclitaxel +/- vinorelbine. A subset analysis of 16 patients with nonmetastatic ULABC Stage IIIB-C (T4N0-3M0) was performed. Among this cohort, 13 patients (81%) underwent planned mastectomy after CRT. RESULTS: Of the 16 patients with Stage IIIB-C disease, acute toxicity included moist desquamation (n = 8) and Grade 3-4 neutropenia (n = 3). Late toxicity included breast reconstruction loss, decreased range of arm motion, lymphedema, and skin toxicity, although none was life-threatening. Of 15 assessable patients, 14 had a clinical response, 7 had a pathologic complete response (pCR) including 6 of 13 patients undergoing mastectomy. With a median follow-up for living patients of 43.8 months, the 4-year actuarial locoregional control, disease-free survival, and overall survival were 83%, 33%, and 56% respectively. CONCLUSIONS: Concurrent WO/WO radiation therapy and paclitaxel +/- vinorelbine is effective locoregional therapy for ULABC with an acceptable toxicity profile. Further investigation of concurrent chemoradiotherapy in ULABC is warranted. PMID- 15752884 TI - Cost-effectiveness of radiation therapy following conservative surgery for ductal carcinoma in situ of the breast. AB - PURPOSE: To assess the cost-effectiveness of radiation therapy (RT) in patients with ductal carcinoma in situ (DCIS) after breast-conserving surgery (BCS). METHODS AND MATERIALS: A Markov model was constructed for a theoretical cohort of 55-year-old women with DCIS over a life-time horizon. Probability estimates for local noninvasive (N-INV), local invasive (INV), and distant recurrences were obtained from National Surgical Adjuvant Breast and Bowel Project (NSABP) B-17. Utilities for eight nonmetastatic health states were collected from both healthy women and DCIS patients. Direct medical (2002 Medicare fee schedule) and nonmedical costs (time and transportation) of RT were ascertained. RESULTS: For BCS + RT vs. BCS alone, the estimated N-INV and INV rates at 12 years were 9% and 8% vs. 16% and 18%, respectively. The incremental cost of adding RT was 3300 US dollars despite an initial RT cost of 8700 US dollars due to higher local recurrence-related salvage costs incurred with the BCS alone strategy. An increase of 0.09 quality-adjusted life-years (QALYs) primarily reflected the lower risk of INV with RT, resulting in an incremental cost-effectiveness ratio (ICER) of 36,700 US dollars/QALY. Sensitivity analyses revealed the ICER to be affected by baseline probability of a local recurrence, relative efficacy of RT in preventing INV, negative impact of an INV on quality of life, and cost of initial RT. Cost of salvage BCS + RT and source of utilities (healthy women vs. DCIS patients) influenced the ICER albeit to a lesser degree. CONCLUSIONS: Addition of RT following BCS for patients with DCIS should not be withheld because of concerns regarding its cost-effectiveness. PMID- 15752885 TI - Feasibility of dose escalation using intensity-modulated radiotherapy in posthysterectomy cervical carcinoma. AB - PURPOSE: To evaluate retrospectively the utility of intensity-modulated radiotherapy (IMRT) in reducing the volume of normal tissues receiving radiation at varying dose levels when the female pelvis after hysterectomy is treated to doses of 50.4 Gy and 54 Gy. METHODS AND MATERIALS: Computed tomography scans from 10 patients who had previously undergone conventional postoperative RT were selected. The clinical tumor volume (vaginal apex and iliac nodes) and organs at risk were contoured. Margins were added to generate the planning tumor volume. The Pinnacle and Corvus planning systems were used to develop conventional and IMRT plans, respectively. Conventional four-field plans were prescribed to deliver 45 Gy (4F(45 Gy)) or 50.4 Gy; eight-field IMRT plans were prescribed to deliver 50.4 Gy (IMRT(50.4 Gy)) or 54 Gy (IMRT(54 Gy)) to the planning tumor volume. All plans were normalized so that > or =97% of the planning tumor volume received the prescribed dose. Student's t test was used to compare the volumes of organs at risk receiving the same doses with different plans. RESULTS: The mean volume of bowel receiving > or =45 Gy was lower with the IMRT(50.4 Gy) (33% lower) and IMRT(54 Gy) (18% lower) plans than with the 4F(45 Gy) plan. The mean volume of rectum receiving > or =45 Gy or > or =50 Gy was also significantly reduced with the IMRT plans despite an escalation of the prescribed dose from 45 Gy with the conventional plans to 54 Gy with IMRT. The mean volume of bladder treated to 45 Gy was the same or slightly lower with the IMRT(50.4 Gy) and IMRT(54 Gy) plans compared with the 4F(45 Gy) plan. Compared with the 4F(45 Gy) plan, the IMRT(50.4 Gy) plan resulted in a smaller volume of bowel receiving 35 45 Gy and a larger volume of bowel receiving 50-55 Gy. Compared with the 4F(45 Gy) plan, the IMRT(54 Gy) plan resulted in smaller volumes of bowel receiving 45 50 Gy; however, small volumes of bowel received 55-60 Gy with the IMRT plan. CONCLUSION: Intensity-modulated RT may permit an increase in the radiation dose that can safely be delivered to the central pelvis and pelvic lymph nodes after hysterectomy. However, dose-volume calculations using individual CT scans do not account for internal organ motion. Detailed data concerning the relationships among radiation dose, treatment volume, and treatment effects are lacking, and prospective studies of pelvic IMRT are needed to determine the safety and efficacy of this treatment. PMID- 15752886 TI - Radical radiotherapy for cervix cancer: the effect of waiting time on outcome. AB - PURPOSE: To assess the effect of treatment waiting time on clinical outcome for patients with cervix cancers treated with radical radiotherapy. METHODS AND MATERIALS: A retrospective analysis was conducted on all cervix cancer patients treated with radical radiotherapy between 1990 and 2001 at the Ottawa Regional Cancer Centre. Analyses were performed according to the three following separate definitions of waiting times: interval from start of radiotherapy to (1) date of initial biopsy, (2) date of examination under anesthesia, and (3) date of radiation oncology consultation. Associations between waiting times and patient characteristics and disease control were investigated using t-tests, analyses of variance, and Cox regression analyses. RESULTS: A total of 195 patients were studied. The vast majority of patients were treated within 5, 6, and 8 weeks of their consultation (91%), examination under anesthesia (88%), and biopsy (81%), respectively. On average, delays between initial biopsy and treatment start were greater for older patients (p = 0.025) (5.8 weeks for <40 years old vs. 6.6 weeks for >70 years old) and those with smaller tumors (p < 0.001) (5.0 weeks for >4 cm vs. 6.3 weeks for < or =4 cm). Univariate analysis revealed no adverse effect of treatment delay on tumor control. Multivariate analysis, with the inclusion of multiple prognostic tumor and treatment parameters, revealed an adverse effect of treatment delay on survival outcomes. CONCLUSIONS: Longer radiotherapy waiting times were found to be associated with diminished survival outcomes for patients treated radically for cervix cancer. The significance of this observed association requires further investigation. PMID- 15752887 TI - Clitoral therapy device for treatment of sexual dysfunction in irradiated cervical cancer patients. AB - PURPOSE: The purpose of this pilot study was to evaluate the efficacy of the clitoral therapy device (Eros Therapy) in alleviating sexual dysfunction in irradiated cervical cancer patients. METHODS AND MATERIALS: Eligible patients had a history of cervical cancer treated with radiotherapy and self-reported sexual dysfunction of sexual arousal and/or orgasmic disorders. Patients used the noninvasive, nonpharmacologic clitoral therapy device using a hand-held, battery powered vacuum to cause clitoral engorgement four times weekly for 3 months during foreplay and self-stimulation. Study instruments included the Female Sexual Function Index, Derogatis Interview for Sexual Functioning, and Dyadic Adjustment Scale. The outcome evaluation was performed at 3 months. RESULTS: Between 2001 and 2002, 15 women were enrolled and 13 completed the study. The median patient age and radiotherapy-enrollment interval was 43.5 years and 2 years, respectively. At baseline, all patients reported symptoms of sexual arousal and/or orgasmic disorders, and some also had sexual desire and pain disorders. At 3 months, statistically significant improvements were seen in all domains tested, including sexual desire, arousal, lubrication, orgasm, sexual satisfaction, and reduced pain. The median Female Sexual Function Index total score increased from 17 to 29.4 (maximal score, 36; p <0.001). The median Derogatis Interview for Sexual Functioning total raw score increased from 46 to 95 (maximal score, 118; p <0.001). At baseline, the Derogatis Interview for Sexual Functioning total T-score corresponded to the bottom 10th percentile of normal sexual functioning. At 3 months, the total T-score placed the patients at the normalcy cutoff. Gynecologic examinations revealed improved mucosal color and moisture and vaginal elasticity and decreased bleeding and ulceration. CONCLUSION: Our results from this pilot study suggest that the clitoral therapy device may alleviate sexual dysfunction in irradiated cervical cancer patients. A randomized, controlled trial is warranted to assess the full benefits of this approach. PMID- 15752888 TI - Pretreatment anemia is correlated with the reduced effectiveness of radiation and concurrent chemotherapy in advanced head and neck cancer. AB - PURPOSE: Pretreatment anemia is an adverse prognostic variable in squamous cell head-and-neck cancer (HNC) patients treated with radiotherapy (RT) alone. Tumor hypoxia is an adverse parameter for treatment with RT alone or with RT and concurrent chemotherapy (CCT). Tumor hypoxia is more prevalent in patients who present with pretreatment hemoglobin (Hgb) concentrations less than 13 g/dL. RT/CCT improves survival over RT alone in advanced HNC, and its use is becoming more widespread. This study was performed to evaluate whether pretreatment Hgb less than 13 g/dL was correlated with treatment outcome in patients with advanced HNC treated with a uniform regimen of RT/CCT. METHODS AND MATERIALS: The study population consisted of patients with AJCC Stage III or IV, M0 HNC who were treated with 70 to 72.5 Gy accelerated hyperfractionated RT (1.25 Gy b.i.d.) and CCT consisting of 2 cycles of CDDP (12-20 mg/m(2)/d x 5 days) and continuous infusion 5-FU (600 mg/m(2)/d x 5 days) during Week 1 and Week 6. A planned break in RT occurred during Week 4. These patients were enrolled on the experimental arm of a prospective randomized trial that compared this regimen to hyperfractionated irradiation alone from 1990 to 1996. RT/CCT was delivered as standard therapy from 1996 to 2000. The primary endpoint was failure-free survival (FFS). Secondary endpoints included local-regional control and overall survival. RESULTS: One hundred and fifty-nine patients were treated from 1990 to 2000. The median (25-75%) pretreatment Hgb was 13.6 (12.2-13.5) g/dL. Hgb was 13 g/dL or higher in 105 patients and less than 13 g/dL in 54 patients. Primary tumor sites included oropharynx (43%), hypopharynx/larynx (36%), oral cavity (9%), and nasopharynx (6%). Seventy-eight percent of the patients with Hgb 13 g/dL or higher and 92% of the patients with Hgb less than 13 g/dL had a primary tumor stage of T3 or T4 (p = 0.01). Node-positive disease was present in 74 of 105 (70%) of patients with Hgb 13 g/dL or higher patients and in 36/54 (67%) of patients with Hgb less than 13 g/dL patients. Median follow-up of surviving patients was 42 months (range, 4-128 months). Five-year FFS was 75% for patients with Hgb 13 g/dL or higher vs. 50% for patients with Hgb less than 13 g/dL had a (p < 0.01). A total of 49 failures occurred in both patient cohorts. The median (25-75%) decrease in Hgb during RT/CCT was 2.2 (1.3-3.1) g/dL, both in patients who failed and in those who remained disease-free. CONCLUSION: Pretreatment Hgb less than 13 g/dL is correlated with adverse outcomes in advanced HNC patients treated with RT/CCT. Whether anemia actually causes poor outcomes remains unknown. The therapeutic effect of anemia correction is being evaluated in prospective trials. PMID- 15752890 TI - Treatment results for nasopharyngeal carcinoma in the modern era: the Hong Kong experience. AB - PURPOSE: To analyze the treatment results achievable for nasopharyngeal carcinoma in the modern era to identify the key failures for future improvement and to provide an updated baseline for future trials. METHODS AND MATERIALS: The results of 2687 consecutive patients treated at all public oncology centers in Hong Kong during 1996-2000 were retrospectively analyzed. The stage distribution (by American Joint Committee on Cancer and International Union Against Cancer staging system, 1997) was 7% Stage I, 41% Stage II, 25% Stage III, and 28% Stage IVA-B. All patients were irradiated with 6-MV photons and the median total dose was 66 Gy. Only 23% of patients had additional treatment with chemotherapy. RESULTS: The 5-year local, nodal, and distant failure-free rates were 85%, 94%, and 81%, respectively; patients with local failure had significantly higher risk of nodal and distant failures. The 5-year progression-free, overall, and cancer-specific survival rates were 63%, 75%, and 80%, respectively. The presenting stage was the most important prognostic factor for all endpoints: with overall survival decreasing from 90% for Stage I to 58% for Stage IVA-B. The results achieved by the 2070 patients treated by radiotherapy alone were almost identical to that of the whole series, the distant failure-free rate among patients with locoregional control was 89% for Stage I-II and 75% for Stage III-IVB. The 860 patients (32%) staged with magnetic resonance imaging achieved significantly better results than those staged by computed tomography, the overall survival being 93% vs. 83% for Stages I-II, and 72% vs. 63% for Stages III-IVB (p = 0.001). CONCLUSIONS: Treatment results for nasopharyngeal carcinoma have substantially improved in the modern era; future trials should be based on updated baseline results. Further reduction of distant failure is important for future breakthrough, particularly for patients with advanced disease. PMID- 15752889 TI - Twice-daily reirradiation for recurrent and second primary head-and-neck cancer with gemcitabine, paclitaxel, and 5-fluorouracil chemotherapy. AB - PURPOSE: We previously demonstrated the efficacy of concurrent gemcitabine, paclitaxel, and 5-fluorouracil in conjunction with twice-daily (1.5-Gy) radiotherapy delivered on alternating weeks (TFGX(2)) in locally advanced head and-neck cancer. Here, we report the clinical outcome and late toxicity of TFGX(2) in a subset of patients previously irradiated to the head and neck. METHODS AND MATERIALS: Twenty-nine previously irradiated patients, presenting with recurrent or second primary head-and-neck cancer, underwent TFGX(2). Twelve patients underwent attempted surgical resection before chemoradiotherapy, 10 of whom were left with no measurable disease. Patients with measurable disease received a median radiation dose of 72 Gy; those with no measurable disease received a median dose of 61 Gy. The cumulative dose ranged from 74.4 to 156.4 Gy (mean, 125.7 Gy; median, 131.0 Gy). RESULTS: The median follow-up was 19.1 months (50.9 months for living patients). The 5-year overall survival rate was 34.5%, and the locoregional control rate was 54.5%. In patients with measurable disease at treatment, the 5-year overall survival and locoregional control rate was 26.3% and 45.1%, respectively, compared with 50.0% (p = 0.14) and 70% (p = 0.31), respectively, for those with no measurable disease. Measurable disease and radiation dose were highly statistically significant for overall survival and locoregional control on multivariate analysis. Of 14 patients assessable for late toxicity, 3 developed Grade 4-5, 8 Grade 2-3, and 3 Grade 0-1 toxicity. CONCLUSION: Aggressive reirradiation with chemotherapy in locally advanced head and-neck cancer provides a chance for long-term cure at the expense of toxicity. Attempted surgical resection before chemoradiotherapy improved disease control and survival. PMID- 15752891 TI - A phase II study of hyperfractionated accelerated radiotherapy (HART) after induction cisplatin (CDDP) and vinorelbine (VNR) for stage III non-small-cell lung cancer (NSCLC). AB - PURPOSE: The purpose was to assess the feasibility and efficacy of hyperfractionated accelerated radiotherapy (HART) after induction chemotherapy for Stage III non-small-cell lung cancer. METHODS AND MATERIALS: Treatment consisted of 2 cycles of cisplatin 80 mg/m(2) on Day 1 and vinorelbine 25 mg/m(2) on Days 1 and 8 every 3 weeks followed by HART, 3 times a day (1.5, 1.8, 1.5 Gy, 4-h interval) for a total dose of 57.6 Gy. RESULTS: Thirty patients were eligible. Their median age was 64 years (range, 46-73 years), 24 were male, 6 were female, 8 had performance status (PS) 0, 22 had PS 1, 9 had Stage IIIA, and 21 had Stage IIIB. All but 1 patient completed the treatment. Common grade > or =3 toxicities during the treatment included neutropenia, 25; infection, 5; esophagitis, 5; and radiation pneumonitis, 3. The overall response rate was 83%. The median survival was 24 months (95% confidence interval [CI], 13-34 months), and the 2-year overall survival was 50% (95% CI, 32-68%). The median progression free survival was 10 months (95% CI, 8-20 months). CONCLUSION: Hyperfractionated accelerated radiotherapy after induction of cisplatin and vinorelbine was feasible and promising. Future investigation employing dose-intensified radiotherapy in combination with chemotherapy is needed. PMID- 15752892 TI - The integration of 18-fluoro-deoxy-glucose positron emission tomography and endoscopic ultrasound in the treatment-planning process for esophageal carcinoma. AB - PURPOSE: Accurate delineation of the gross tumor volume (GTV) is important in radiation therapy treatment planning. We evaluated the impact of PET and endoscopic ultrasound (EUS) compared with CT simulation in the planning of radiation fields for patients with esophageal carcinoma. MATERIAL AND METHODS: Twenty-five patients presenting with esophageal carcinoma for radiation therapy underwent PET scans in the treatment position after conventional CT simulation. Patients underwent PET/CT scanning after being injected with 10 to 20 mCi of [F 18]-2-deoxy-2-fluro-D-glucose. The length of the abnormality seen on the CT portion of the PET/CT scan vs. the PET scan alone was determined independently by 2 separate investigators. The length of the GTV and detection of regional adenopathy by PET was also correlated with EUS in 18 patients. Of the 18 patients who had EUS, 2 had T2 tumors and 16 had T3 tumors. Eighteen patients had adenocarcinoma and 7 had squamous cell carcinoma. Nine tumors were located at the gastroesophageal junction, 8 at the lower esophagus, 7 in the middle esophagus, and 1 in the cervical esophagus. The PET scans were reviewed to determine the length of the abnormality by use of a standard uptake value (SUV) of 2.5 to delineate the tumor extent. RESULTS: The mean length of the cancer was 5.4 cm (95% CI 4.4-6.4 cm) as determined by PET scan, 6.77 cm (95% CI, 5.6-7.9 cm) as determined by CT scan, and 5.1 cm (95% CI, 4.0-6.1 cm) for the 22 patients who had endoscopy. The length of the tumors was significantly longer as measured by CT scans compared with PET scans (p = 0.0063). EUS detected significantly more patients with periesophageal and celiac lymphadenopathy compared to PET and CT. The SUV of the esophageal tumors was higher in patients with peri-esophageal lymphadenopathy identified on PET scans. CONCLUSION: Endoscopic ultrasound and PET scans can add additional information to aid the radiation oncologist's ability to precisely identify the GTV in patients with esophageal carcinoma. PMID- 15752893 TI - Assessment of quality of life in patients with rectal cancer treated by preoperative radiotherapy: a longitudinal prospective study. AB - PURPOSE: To assess prospectively the quality of life (QOL) of patients treated by preoperative radiotherapy (RT) and surgery for locally advanced rectal cancer. METHODS AND MATERIALS: We studied 53 patients treated with bi-fractionated RT (50 Gy in 40 fractions within 4 weeks) followed at a median interval of 45 days by abdominoperineal resection in 11 patients and low anterior resection in 42 patients. Their QOL was assessed using two self-rating questionnaires developed by the European Organization for Research and Treatment of Cancer (EORTC): one was cancer specific (EORTC QLQ-C30) and one was site specific (EORTC QLQ-C38). The questionnaires were completed before RT and 12-16 months after RT, at which time 17 patients had undergone colostomy. We hypothesized that at least some scores of the various scales would vary between the two analyses. RESULTS: Compared with the pre-RT scores, at 1 year, patients reported statistically significant improvement in their emotional state (median 75 vs. 100, p <0.0001), perspective of the future (67 vs. 100, p = 0.0004), and their global QOL (75 vs. 83, p = 0.0008), as well as a decrease in GI symptoms (13 vs. 0, p = 0.002). However, the sexual dysfunction score increased significantly, particularly in men (17 vs. 83, p = 0.0045), and a trend toward a lower body image score was observed (100 vs. 89, p = 0.068). At 1 year, patients with colostomies reported similar or significantly improved symptom scores for fatigue, pain, GI problems, and sleep disturbance, but no such improvements were observed in patients without stomas. CONCLUSION: One year after combined treatment for locally advanced rectal cancer, patients exhibited statistically significant improvement in some important QOL outcomes, including global QOL, despite a decrease in sexual function and body image. Any additional improvement in QOL outcome may require refinements in the RT and surgical techniques to reduce late sequelae, particularly sexual dysfunction. Our results suggest that QOL considerations do not justify sphincter-conserving approaches if locoregional tumor control would be compromised. PMID- 15752894 TI - Radiochemotherapy in the conservative treatment of anal canal carcinoma: retrospective analysis of results and radiation dose effectiveness. AB - PURPOSE: This retrospective analysis reports the results on patients with anal canal carcinoma treated by combined radiotherapy and chemotherapy. METHODS AND MATERIALS: Between March 1993 and December 2001, 43 patients with anal canal carcinoma were treated with radiochemotherapy at the Hospital do Cancer A.C. Camargo. Stage distribution was as follows: I, 3 (7%); II, 23 (53.5%); IIIA, 8 (18.6%); and IIIB, 9 (21%). The median age was 56 years (range, 36-77 years) with most patients being women (4:1). External radiotherapy (RT) was delivered at the whole pelvis followed by a boost at the primary tumor. The median dose of RT at the whole pelvis and at the primary tumor was 45 Gy and 55 Gy, respectively. Chemotherapy was carried out during the first and last 4 days of RT with continuous infusion of 5-fluorouracil (1000 mg/m(2)) and bolus mitomycin C (10 mg/m(2)). Median overall treatment time was 51 days (range, 30-129 days). Thirty four patients (79%) did not receive elective RT at the inguinal region. Patient's age, tumor stage, overall treatment time, and RT dose at primary tumor were variables analyzed for survival and local control. RESULTS: Median follow-up time was 42 months (range, 4-116 months). Overall survival and colostomy-free survival at 5 years was 68% and 52%, respectively. Overall survival according to clinical stage was as follows: I, 100%; II, 82%; IIIA, 73%; and IIIB, 18% (p = 0.0049). Complete response was observed in 40 patients (93%). Local recurrence occurred in 9 (21%) patients, and of these, 6 were rescued by surgery. Local control with a preserved sphincter was observed in 34 patients (79%). According to the RT dose, local control was higher among patients who received more than 50 Gy at primary tumor (86.5% vs. 34%, p = 0.012). Inguinal failure was observed in 5 patients (15%) who did not receive inguinal elective RT. Distant metastasis was observed in 11 patients (25.6%). Temporary interruption of the treatment as a result of acute toxicity was necessary in 12 patients (28%). Four patients developed mild chronic complications. CONCLUSIONS: This analysis suggests that the treatment scheme employed was effective for anal sphincter preservation and local control; however, the incidence of distant metastases was relatively high. The clinical stage was the main prognostic factor for overall survival. Local control was higher in patients treated with doses of more than 50 Gy at primary tumor. The high incidence of inguinal failure implies the need for elective RT in this region. PMID- 15752895 TI - Local radiotherapy for patients with unresectable hepatocellular carcinoma. AB - PURPOSE: To evaluate the response to local radiotherapy (RT) for unresectable hepatocellular carcinoma (HCC) and to analyze the dose-response relationship and the treatment-related morbidities. METHODS AND MATERIALS: Between 1998 and 2002, 59 patients who were treated with localized RT were evaluated. RT was delivered with a curative intent, and the radiation dose was 30-55 Gy (biologic effective dose of 39.0-70.2 Gy(10) using the alpha/beta ratio of 10 Gy) with 2-3 Gy as a daily dose. The tumor response was evaluated by the change in maximum tumor size on serial CT scans, and the morbidity was evaluated by the Common Terminology Criteria for Adverse Events v3.0. RESULTS: An objective tumor response was achieved in 39 of 59 patients (66.1%) with complete response (CR) in 5 patients and partial response (PR) in 34 patients. More than 50 Gy(10) had a significant response; CR or PR was 72.8% with >50 Gy(10) and 46.7% with < or =50 Gy(10) (p = 0.0299). The 2-year overall survival rate after RT was 27.4% (median survival time: 10 months), and this was affected by the tumor response (p = 0.0640); the 2 year overall survival rate after RT was 50.0% for CR and 21.8% for PR. There was no Grade 3 or 4 acute toxicity, and 3 patients (5.1%) developed gastric or duodenal ulcer. CONCLUSIONS: Radiotherapy for unresectable HCC resulted in 66.1% of tumor response with acceptable toxicity, and the radiation dose seems to be a significant prognostic factor in RT response for HCC. PMID- 15752896 TI - Combination of gamma-radiation antagonizes the cytotoxic effects of vincristine and vinblastine on both mitotic arrest and apoptosis. AB - PURPOSE: Combination therapy with different modalities is a common practice in the treatment of cancer. The promising clinical profile of vincristine and vinblastine has promoted considerable interest in combining these vinca alkaloids with radiation therapy to treat a variety of solid tumors. However, the therapeutic efficacy and the interaction between the vinca alkaloids with radiation is not entirely clear. In this study, we assessed the potential interactions in the combination of vincristine or vinblastine with gamma radiation against human tumor cells in vitro. METHODS AND MATERIALS: Vincristine or vinblastine and gamma-radiation were administrated at three different sequences designed as preradiated, coradiated, and postradiated combinations in human breast cancer cells and human epidermoid carcinoma cells. The cytotoxic interactions and mutual influences between these two modalities were analyzed by a series of assays including cytotoxic, morphologic, and biochemical examinations. RESULTS: Our results showed that the combination of these two modalities did not produce any synergistic or additive effects. Instead, the clonogenic assays showed the survival rates of these combinations were increased up to 2.17-fold and 2.7-fold, respectively, of those treated with vincristine or vinblastine alone (p < 0.01). DNA fragmentation, TalphaT-mediated dUTP nick end labeling (TUNEL) assay, and flow cytometric assays also showed that the combination of gamma-radiation significantly interfered with the ability of these vinca alkaloids to induce apoptosis. Further analyses indicated that addition of gamma-radiation resulted in cell cycle arrest at the G(2) phase, which subsequently prevented the mitotic arrest induced by vincristine or vinblastine. In addition, biochemical examinations revealed that gamma-radiation regulated p34(cdc2)/cyclin B1 and survivin, and inhibited IkappaBalpha degradation and bcl 2 phosphorylation. CONCLUSIONS: These results suggest that gamma-radiation might specifically block the cell cycle at the G(2) phase, which in turn interferes with the cytotoxic effects of vincristine or vinblastine on mitotic arrest and apoptosis. Thereby, it eventually results in an antagonistic interaction between these two modalities. This finding may be implicated in the clinical application of combination therapy of vinca alkaloids and radiation. PMID- 15752897 TI - Protective effect of superoxide dismutase in radiation-induced intestinal inflammation. AB - PURPOSE: To analyze the therapeutic value of Cu/Zn-superoxide dismutase (SOD1) supplementation in an experimental model of radiation-induced intestinal inflammation and explore its mechanistic effects. METHODS AND MATERIALS: Mice were subjected to abdominal irradiation with 10 Gy or sham irradiation and studied 24 or 72 hours after radiation. Groups of mice were treated with 0.1, 4, or 6 mg/kg/day of SOD1 or vehicle. Leukocyte-endothelial cell interactions in intestinal venules were assessed by intravital microscopy. Endothelial intercellular adhesion molecule-1 (ICAM-1) expression was determined with radiolabeled antibodies. Effects of SOD1 on histologic damage and levels of lipid hydroperoxides were also measured. RESULTS: A significant increase in the flux of rolling leukocytes and number of firmly adherent leukocytes in intestinal venules was observed at 24 and 72 hours after irradiation. Treatment with SOD1 had no effect on leukocyte rolling but significantly and dose-dependently decreased firm leukocyte adhesion to intestinal venules. Treatment with SOD1 at doses that reduced leukocyte recruitment abrogated the increase in hydroperoxides in intestinal tissue and ICAM-1 upregulation in intestinal endothelial cells. The inflammatory score, but not a combined histology damage score, was also significantly reduced by SOD1. CONCLUSIONS: Treatment with SOD1 decreases oxidative stress and adhesion molecule upregulation in response to abdominal irradiation. This is associated with an attenuation of the radiation-induced intestinal inflammatory response. PMID- 15752898 TI - Post-irradiation phosphorylation of structural maintenance chromosome 1 (SMC1) is independent of the Fanconi protein pathway. AB - PURPOSE: To confirm the sensitivity of cells from patients with Fanconi anemia (FA) to ionizing radiation, and to determine whether the phosphorylation of structural maintenance chromosome 1 (SMC1) was associated with radiosensitivity, as it is in other DNA repair disorders. METHODS AND MATERIALS: Using lymphoblastoid cell lines from FA patients before and after exposure to ionizing radiation, the colony survival assay, radioresistant DNA synthesis, and SMC1 phosphorylation were measured. FA lymphoblastoid cell lines that had been transfected with the wild-type FANC gene were used as controls. RESULTS: Cells from FA patients of six complementation groups were radiosensitive. Despite this, SMC1 phosphorylation was normal in each case; radioresistant DNA synthesis, a measure of S phase checkpoint integrity, was defective in FANCD2 lymphoblastoid cell lines and was corrected in FANCD2 + D2 cells. CONCLUSIONS: The data indicate that the FANC pathway proteins play a major role in the cellular responses to ionizing radiation, but not in SMC1 phosphorylation or in the S phase checkpoint of FANCD2-deficient cells. Thus, SMC1 activation is not a common denominator of radiosensitivity, as has been suggested by radiation responses of cells from ataxia-telangiectasia, Nijmegen breakage syndrome, or Mre11 deficiency patients. PMID- 15752899 TI - Enhanced delivery of iodine for synchrotron stereotactic radiotherapy by means of intracarotid injection and blood-brain barrier disruption: quantitative iodine biodistribution studies and associated dosimetry. AB - PURPOSE: Synchrotron stereotactic radiotherapy (SSR) is a binary cancer treatment modality that involves the selective accumulation of a high Z element, such as iodine, in tumors, followed by stereotactic irradiation with kilovoltage X-rays from a synchrotron source. The success of SSR is directly related to the absolute amount of iodine achievable in the tumor. The purposes of this preclinical study were to determine whether the delivery of iodine to brain tumor models in rats could be enhanced by the means of its intracarotid injection with or without a hyperosmotic solution and to evaluate corresponding absorbed X-ray doses. METHODS AND MATERIALS: Experiments were performed on four groups of F98 glioma-bearing rats, which received either intracarotid (IC) or intravenous (IV) infusions of a mixture (6 mL in 12 min) of an iodinated contrast agent associated or not with a transient blood-brain barrier opener (mannitol). The mixture volumetric proportions were 8/13 of Iomeron (C = 350 mg/mL) for 5/13 of mannitol or saline, respectively. Absolute iodine concentration kinetic was measured in vivo in the tumor, blood, contralateral and ipsilateral brain, and muscle by monochromatic computed tomography. Associated dosimetry was performed by computing the iodine dose enhancement factor (DEF) in each region and building dose distribution maps by analytical simulations. RESULTS: Infusion of mannitol significantly enhanced iodine tumor uptake compared with the control values (p < 0.0001 and p = 0.0138, for IC and IV protocols, respectively). The mean iodine concentrations (C) reached 20.5 +/- 0.98 mg/mL (DEF = 4.1) after administration of iodine and mannitol vs. 4.1 +/- 1.2 mg/mL i.c. with serum (DEF = 1.6). The tumor iodine uptakes after jugular injection with mannitol (C = 4.4 +/- 2.1 mg/mL, DEF = 1.7) were not significantly different from IC injection of iodine without mannitol (p = 0.8142). The IV injection of iodine with saline led to an iodine concentration in the tumor of 1.2 +/- 0.98 mg/mL and a DEF of 1.2. CONCLUSIONS: This study established that optimizing the delivery of iodine by means of IC injection combined with a blood-brain barrier opener (mannitol) significantly increases the iodine uptake of F98 rat gliomas. This infusion protocol could potentially enhance the efficacy of SSR treatment, because the radiation dose is proportional to the iodine amount present in the irradiation bed. PMID- 15752900 TI - Inhibition of phosphatidylinositol-3-kinase causes cell death through a protein kinase B (PKB)-dependent mechanism and growth arrest through a PKB-independent mechanism. AB - PURPOSE: To identify whether inhibition of phosphatidylinositol-3-kinase (PI3K) causes apoptosis through inhibition of protein kinase B (PKB), implicating PKB as an important therapeutic target in prostate cancer. METHODS AND MATERIALS: After treatment with the PI3K inhibitor, LY294002, proliferation and apoptosis of the prostate cancer cell line, LNCaP, were measured by cell cycle analysis and cleavage of poly (ADP-ribose) polymerase. To test the hypothesis that inhibition of PKB is responsible for the LY294002-induced apoptosis, LNCaP cells expressing a constitutively active form of PKB were generated. RESULTS: Treatment of LNCaP cells with the PI3K inhibitor, LY294002, caused inactivation of PKB, growth arrest, and apoptosis. LY294002-induced apoptosis was increased in the absence of serum. The G1 growth arrest was associated with an increase in p27(kip1) expression. Cells expressing constitutively active PKB were protected from apoptosis induced by LY294002, but not from the G1 growth arrest induced by PI3K inhibition. CONCLUSION: These data suggest that PKB activity regulates apoptosis, but not G1 arrest, and identify PKB as a potential critical target for cancer therapy. Targeted therapy against kinases might complement more conventional therapies, including androgen suppression for prostate cancer. PMID- 15752902 TI - Characterization of carbonic anhydrase IX (CA IX) as an endogenous marker of chronic hypoxia in live human tumor cells. AB - PURPOSE: Published clinical studies provide conflicting data regarding the prognostic significance of carbonic anhydrase IX (CA IX) overexpression as an endogenous marker of tumor hypoxia and its comparability with other methods of hypoxia detection. We performed a systematic analysis of CA IX protein levels under various in vitro conditions of tumor hypoxia in HT 1080 human fibrosarcoma and FaDu human pharyngeal carcinoma cells. Because sorting of live CA IX positive cells from tumors provides a tool to study the radiosensitivity of chronically hypoxic cells, we modified and tested a CA IX flow cytometry protocol on mixed hypoxic/aerobic suspensions of HT 1080 and FaDu cells. METHODS AND MATERIALS: HT 1080 and FaDu cells were treated with up to 24 h of in vitro hypoxia and up to 96 h of reoxygenation. To test the effect of nonhypoxic stimuli, glucose and serum availability, pH and cell density were modified. CA IX protein was quantified in Western blots of whole-cell lysates. Mixed suspensions with known percentages of hypoxic cells were prepared for CA IX flow cytometry. The same mixtures were assayed for clonogenic survival after 10 Gy. RESULTS: Hypoxia-induced CA IX protein expression was seen after >6 h at < or =5% O(2), and protein was stable over 96 h of reoxygenation in both cell lines. Glucose deprivation abolished the hypoxic CA IX response, and high cell density caused CA IX induction under aerobic conditions. Measured percentages of CA IX-positive cells in mixtures closely reflected known percentages of hypoxic cells in HT 1080 and were associated with radioresistance of mixtures after 10 Gy. CONCLUSION: CA IX is a stable marker of current or previous chronic hypoxia but influenced by nonhypoxic stimuli. Except the time course of accumulation, all properties of this marker resembled our previous findings for hypoxia-inducible factor-1alpha. A modified flow cytometry protocol provided good separability of CA IX-negative and positive cells in vitro and can be applied to sort chronically hypoxic cells from tumors. PMID- 15752901 TI - Expression of bone morphogenic protein 2/4, transforming growth factor-beta1, and bone matrix protein expression in healing area between vascular tibia grafts and irradiated bone-experimental model of osteonecrosis. AB - PURPOSE: For the surgical treatment of osteoradionecrosis after multimodal therapy of head-and-neck cancers, free vascular bone grafts are used to reconstruct osseous structures in the previously irradiated graft bed. Reduced, or even absent osseous healing in the transition area between the vascular graft and the irradiated graft bed represents a clinical problem. Inflammatory changes and fibrosis lead to delayed healing, triggered by bone morphogentic protein 2/4 (BMP2/4) and transforming growth factor (TGF)-beta(1). Given the well-known fibrosis-inducing activity of TGF-beta(1), an osteoinductive effect has been reported for BMP2/4. However, the influence of irradiation (RT) on this cytokine expression remains elusive. Therefore, the aim of the present in vivo study was to analyze the expression of BMP2/4, TGF-beta(1), collagen I, and osteocalcin in the transition area between the bone graft and the graft bed after RT. METHODS AND MATERIALS: Twenty Wistar rats (male, weight 300-500 g) were used in this study. A free vascular tibia graft was removed in all rats and maintained pedicled in the groin region. Ten rats underwent RT with 5 x 10 Gy to the right tibia, the remainder served as controls. After 4 weeks, the previously removed tibia grafts were regrafted into the irradiated (Group 1) and nonirradiated (Group 2) graft beds. The interval between RT and grafting was 4 weeks. After a 4 week osseous healing period, the bone grafts were removed, and the transition area between the nonirradiated graft and the irradiated osseous graft bed was examined histomorphometrically (National Institutes of Health imaging program) and immunohistochemically (avidin-biotin-peroxidase complex) for the expression of BMP2/4, TGF-beta(1), collagen I, and osteocalcin. RESULTS: Absent or incomplete osseous healing of the graft was found in 9 of 10 rats after RT with 50 Gy and in 1 of 10 of the rats with nonirradiated osseous grafts. Histomorphometrically, the proportion of osseous healing in the transition area was 17% in Group 1 and 48% in Group 2 (p = 0.001). Compared with the nonirradiated rats, reduced enchondral and perichondral ossification was found in the healing area after RT, with a reduction of BMP2/4 and osteocalcin expression. TGF-beta(1) and collagen I expression in the transition area to the irradiated osseous graft bed was significantly increased compared with that in the nonirradiated osseous graft bed. CONCLUSION: After RT, osseous healing of vascular bone grafts is significantly reduced and may be a result of radiation induced inhibition of BMP2/4 and osteocalcin expression. In addition, induction of TGF-beta(1) and collagen I expression occurs. Because the effects of the TGF beta superfamily are manifold and partially unknown, additional research directions could be in the exogenous application of BMP2/4 and inhibition of TGF beta(1) by antibody treatment to search for appropriate therapeutic approaches for improving osseous healing in the irradiated graft bed. PMID- 15752903 TI - Intravascular contrast agent-enhanced MRI measuring contrast clearance and tumor blood volume and the effects of vascular modifiers in an experimental tumor. AB - PURPOSE: To examine the feasibility of using the MRI blood pool agent NC100150 for evaluation of tumor blood volume (TBV) estimates by both dynamic contrast enhanced MRI (DCE-MRI) and susceptibility contrast MRI assays in an experimental tumor. Contrast agent clearance (K(trans); depends on perfusion and permeability) from the DCE-MRI time curves was estimated, and changes in TBV and K(trans) were measured after administration of two drugs that reduce perfusion by different mechanisms. METHODS AND MATERIALS: The DCE-MRI experiments were simulated with expected physiologic values for the C3H mouse mammary carcinoma. The C3H tumor was examined by DCE-MRI and susceptibility contrast MRI with NC100150 (NC100150 Injection, Clariscan; Amersham Health, Oslo, Norway) after treatment with either hydralazine or combretastatin (Oxigene, Boston, MA). RESULTS: Simulations showed that reliable estimates of changes in TBV and K(trans) could be performed with DCE-MRI. Hydralazine was shown to reduce TBV as measured by both assays and to reduce K(trans). Dynamic contrast-enhanced MRI also suggested that TBV and K(trans) were reduced in combretastatin-treated tumors, and the TBV reduction was confirmed by susceptibility contrast MRI. Data suggested the drug to affect mainly the total TBV, whereas microvessels as such seemed less altered. CONCLUSION: The study supports the use of the combined DCE-MRI and susceptibility contrast MRI assay with a blood pool agent in characterizing tumors and their response to treatment. PMID- 15752904 TI - Combining vascular and cellular targeting regimens enhances the efficacy of photodynamic therapy. AB - PURPOSE: Photodynamic therapy (PDT) can be designed to target either tumor vasculature or tumor cells by varying the drug-light interval. Photodynamic therapy treatments with different drug-light intervals can be combined to increase tumor response by targeting both tumor vasculature and tumor cells. The sequence of photosensitizer and light delivery can influence the effect of combined treatments. METHODS AND MATERIALS: The R3327-MatLyLu rat prostate tumor model was used in this study. Photosensitizer verteporfin distribution was quantified by fluorescence microscopy. Tumor blood flow changes were monitored by laser-Doppler system and tumor hypoxia was quantified by the immunohistochemical staining for the hypoxic marker EF5. The therapeutic effects of PDT treatments were evaluated by the histologic examination and tumor regrowth assay. RESULTS: Fluorescence microscopic studies indicated that tumor localization of verteporfin changed from predominantly within the tumor vasculature at 15 min after injection, to being throughout the tumor parenchyma at 3 h after injection. Light treatment (50 J/cm(2)) at 15 min after verteporfin injection (0.25 mg/kg, i.v.) induced significant tumor vascular damage, as manifested by tumor blood flow reduction and increase in the tumor hypoxic fraction. In contrast, the vascular effect observed after the same light dose (50 J/cm(2)) delivered 3 h after administration of verteporfin (1 mg/kg, i.v.) was an initial acute decrease in blood flow, followed by recovery to the level of control. The EF5 staining revealed no significant increase in hypoxic fraction at 1 h after PDT using 3 h drug-light interval. The combination of 3-h interval PDT and 15-min interval PDT was more effective in inhibiting tumor growth than each individual PDT treatment. However, it was found that the combined treatment with the sequence of 3-h interval PDT before 15-min interval PDT led to a superior antitumor effect than the other combinative PDT treatments. Histologic studies confirmed that this combined treatment led to damage to both tumor vasculature and tumor cells. Importantly, the combined PDT treatment did not increase normal tissue damage and tissue recovered well at 60 days after treatment. CONCLUSIONS: Our results suggest that targeting both tumor vascular and cellular compartments by combining a long-interval PDT with a short-interval PDT can be an effective and safe way to enhance PDT damage to tumor tissue. PMID- 15752905 TI - Improved treatment planning for COMS eye plaques. AB - PURPOSE: A recent reanalysis of the Collaborative Ocular Melanoma Study (COMS) medium tumor trial concluded that incorporating factors to account for anisotropy, line source approximation, the gold plaque, and attenuation in the Silastic seed carrier into the dose calculations resulted in a significant and consistent reduction of calculated doses to structures of interest within the eye. The authors concluded that future eye plaque dosimetry should be "performed using the most up-to-date parameters available." The reason these factors are important is attributable to the low energy (125)I radiation (approximately 28 keV) that is primarily absorbed by the photoelectric process. Photoelectric absorption is quite dependent on the atomic composition of the absorbing material. Being 40% silicon by weight, the effective atomic number of Silastic is significantly greater than that of water. Although the AAPM TG43 brachytherapy formalism inherently addresses the issues of source anisotropy and geometry, its parameter that accounts for scatter and attenuation, the radial dose function g(r), assumes that the source is immersed in infinite homogeneous water. In this work, factors are proposed for (125)I that correct for attenuation in the Silastic carrier and scatter deficits resulting from the gold plaque and nearby air. The implications of using (103)Pd seeds in COMS plaques are also discussed. METHODS AND MATERIALS: An existing TG43-based ophthalmic plaque planning system was modified to incorporate additional scatter and attenuation correction factors that better account for the path length of primary radiation in the Silastic seed carrier and the distance between the dose calculation point and the eye-air interface. RESULTS: Compared with homogeneous water, the dose-modifying effects of the Silastic and gold are greatest near the plaque surface and immediately adjacent to the plaque, while being least near the center of the eye. The calculated dose distribution surrounding a single (125)I seed centered in a COMS 20 mm plaque was found to be consistent with previously published examples that used thermoluminescent dosimetry measurements and Monte Carlo methods. For fully loaded 12 and 20 mm plaques, calculated dose to critical ocular structures ranged from 16%-50% less than would have been reported using the standard COMS dose calculation protocol. CONCLUSIONS: Treatment planning for COMS eye plaques that accurately accounts for the presence of the gold, Silastic and extraocular air is both possible and practical. PMID- 15752907 TI - Dosimetric advantages of IMRT simultaneous integrated boost for high-risk prostate cancer. AB - PURPOSE: A sequential two-phase process, initial and boost irradiation, is the common practice for the radiotherapy management of high-risk prostate cancer. In this work, we explore the feasibility of using intensity modulated radiation therapy (IMRT) simultaneous integrated boost (SIB), a single-phase process, to simultaneously deliver high dose to the prostate and lower dose to the pelvic nodes. In addition, we introduce the concept of voxel-equivalent dose for the comparison of treatment plans. METHODS AND MATERIALS: The SIB is designed to deliver the same dose (e.g., 45 Gy, 25 x 1.8 Gy) as the conventional method to the pelvic nodes and to deliver higher doses to prostate in the same 25 fractions (i.e., hypofractionation). The equivalent uniform dose (EUD) was used to determine suitable SIB fractionations that deliver the biologically equivalent doses to prostate. For tumor, the EUD was estimated based on the linear quadratic (LQ) model. The most recent LQ parameters derived from clinical data for prostate cancer were used. The sensitivity of LQ parameters was evaluated. The EUD for normal tissue was computed based on the widely used Lyman model. To be able to consider biologic effectiveness spatially (e.g., voxel by voxel), we propose a new concept, termed the voxel-equivalent dose (VED). The calculation of VED was similar to that for EUD, except that it was done within a voxel. To demonstrate dosimetric feasibility and advantages of the proposed IMRT SIB, we have performed a retrospective planning study on selected patient cases using commercial IMRT and three-dimensional (3D) planning systems. Four treatment scenarios were considered: (1) the conventional 3D plan for initial whole-pelvic irradiation and subsequent conventional 3D boost plan for prostate gland, (2) the conventional 3D plan for initial whole-pelvic irradiation and subsequent IMRT boost plan for prostate, (3) IMRT plan for initial whole-pelvic irradiation and subsequent IMRT boost plan for prostate, and (4) IMRT SIB. EUDs and VED-based dose-volume histograms for prostate, pelvic nodes, small bowel, rectum, bladder, and other tissue for all 4 scenarios were compared. RESULTS: A series of equivalent hypofractionation regimens suitable for the IMRT SIB were obtained for high-risk prostate cancer. For example, the conventional treatment regimen of 42 x 1.8 Gy (EUD = 75.4 Gy) would be equivalent to a SIB regimen of 25 x 2.54 Gy. From the comparison of 3D VED dose distributions and dose-volume histograms between the SIB and the conventional two-phase irradiation, we found that the SIB offers better or equivalent dose conformity to prostate and pelvic nodes and better sparing to the critical structures. For example, for the 4 treatment scenarios with a prostate EUD of 75.4 Gy, the corresponding rectal EUDs are 67.1 (3D + 3D), 65.6 Gy (3D + IMRT), 63.7 Gy (IMRT + IMRT), and 62.0 Gy (SIB). CONCLUSIONS: A new IMRT simultaneous integrated boost strategy that irradiates prostate via hypofractionation while irradiating pelvic nodes with the conventional fractionation is proposed for high-risk prostate cancer. Compared to the conventional two-phase treatment, the proposed SIB technique offers potential advantages, including better sparing of critical structures, more efficient delivery, shorter treatment duration, and better biologic effectiveness. PMID- 15752906 TI - MRI-based volumetric assessment of cardiac anatomy and dose reduction via active breathing control during irradiation for left-sided breast cancer. AB - PURPOSE: Heart dose-volume analysis using computed tomography (CT) is limited because of motion artifact and poor delineation between myocardium and ventricular space. We used dedicated cardiac magnetic resonance imaging (MRI) to quantify exclusion of left ventricular (LV) myocardium via active breathing control (ABC) during left breast irradiation and to determine the correlation between irradiated whole heart and LV volumes. METHODS AND MATERIALS: Fifteen patients who completed adjuvant irradiation for early-stage left breast cancer participated. Treatment consisted of 45 Gy to the entire breast using ABC followed by a 16-Gy electron boost to the lumpectomy cavity. Patients underwent planning CT scans in free breathing (FB) and moderate deep inspiration breath hold (mDIBH). Electrocardiogram-gated cardiac MRI was performed in the treatment position using alpha-cradle immobilization. MRI scans were acquired in late diastole (LD), mid-diastole (MD), and systole (S) for both FB and mDIBH. After image fusion with the patients' radiation therapy planning CT scan, MRI LV volumes were defined for the three examined phases of the cardiac cycle, and comparative dose-volume analysis was performed. RESULTS: Cardiac volume definition was found to differ significantly because of combinations of respiratory and intrinsic heart motion. The fraction of LV myocardium receiving 50% (22.5 Gy) of the prescribed whole breast dose (V(22.5)) was reduced by 85.3%, 91.8%, and 94.6% via ABC for LD, MD, and S, respectively. Linear regression revealed strong correlation between MRI-defined whole heart and LV V(22.5) reduction via ABC, suggesting that LV myocardium accounts for up to approximately 50% of the excluded heart volume through this technique. Significant but weaker correlations were noted between CT-defined whole heart and LV V(22.5) reductions with marked variability in the measurements of patients with larger amounts of heart in the treatment field. CONCLUSIONS: Cardiac MRI demonstrated a significant reduction in LV myocardium irradiated with the use of ABC. The correlation between reduction in V(22.5) values for LV wall and CT-defined whole heart suggests that CT is adequate for determining which patients are likely to benefit from ABC treatment, but inaccuracies inherent to standard CT dictate that more detailed imaging studies such as MRI are required for accurate cardiotoxicity assessment. PMID- 15752908 TI - Use of deformed intensity distributions for on-line modification of image-guided IMRT to account for interfractional anatomic changes. AB - PURPOSE: Recent imaging studies have demonstrated that there can be significant changes in anatomy from day to day and over the course of radiotherapy as a result of daily positioning uncertainties and physiologic and clinical factors. There are a number of strategies to minimize such changes, reduce their impact, or correct for them. Measures to date have included improved immobilization of external and internal anatomy or adjustment of positions based on portal or ultrasound images. Perhaps the most accurate way is to use CT image-guided radiotherapy, for which the possibilities range from simple correction of setup based on daily CT images to on-line near real-time intensity modulated radiotherapy (IMRT) replanning. In addition, there are numerous intermediate possibilities. In this paper, we report the development of one such intermediate method that takes into account anatomic changes by deforming the intensity distributions of each beam based on deformations of anatomy as seen in the beam's eye-view. METHODS AND MATERIALS: The intensity distribution deformations are computed based on anatomy deformations discerned from the changes in the current image relative to a reference image (e.g., the pretreatment CT scan). First, a reference IMRT plan is generated based on the reference CT image. A new CT image is acquired using an in-room CT for every fraction. The anatomic structure contours are obtained for the new image. (For this article, these contours were manually drawn. When image guided IMRT methods are implemented, anatomic structure contours on subsequent images will likely be obtained with automatic or semiautomatic means. This could be achieved by, for example, first deforming the original CT image to match today's image, and then using the same deformation transformation to map original contours to today's image.) The reference intensity distributions for each beam are then deformed so that the projected geometric relationship within the beam's-eye-view between the anatomy (both target and normal tissues) extracted from the reference image and the reference intensity distribution is the same as (or as close as possible to) the corresponding relationship between anatomy derived from today's image and the newly deformed intensity distributions. To verify whether the dose distributions calculated using the deformed intensity distributions are acceptable for treatment as compared to the original intensity distributions, the deformed intensities are transformed into leaf sequences, which are then used to compute intensity and dose distributions expected to be delivered. The corresponding dose volume histograms and dose-volume and dose-response indices are also computed. These data are compared with the corresponding data derived (a) from the original treatment plan applied to the original image, (b) from the original treatment plan applied to today's image, and (c) from a new full-fledged IMRT plan designed based on today's image. RESULTS: Depending on the degree of anatomic changes, the use of an IMRT plan designed based on the original planning CT for the treatment of the current fraction could lead to significant differences compared to the intended dose distributions. CT-guided setup compared to the setup based on skin marks or bony landmarks may improve dose distributions somewhat. Replanning IMRT based on the current fraction's image yields the best physically deliverable plan (the "gold standard"). For the prostate and head-and-neck examples studied as proof of principle, the results of deforming intensities within each beam based on the anatomy seen in the beam's-eye-view are a good approximation of full fledged replanning compared with other alternatives. CONCLUSIONS: Our preliminary results encourage us to believe that deforming intensities taking into account deformation in the anatomy may be a rapid way to produce new treatment plans on line in near real-time based on daily CT images. The methods we have developed need to be applied to a group of patients for both prostate and head-and-neck cases to confirm the validity of our approach. PMID- 15752909 TI - Inverse treatment planning based on MRI for HDR prostate brachytherapy. AB - PURPOSE: To develop and optimize a technique for inverse treatment planning based solely on magnetic resonance imaging (MRI) during high-dose-rate brachytherapy for prostate cancer. METHODS AND MATERIALS: Phantom studies were performed to verify the spatial integrity of treatment planning based on MRI. Data were evaluated from 10 patients with clinically localized prostate cancer who had undergone two high-dose-rate prostate brachytherapy boosts under MRI guidance before and after pelvic radiotherapy. Treatment planning MRI scans were systematically evaluated to derive a class solution for inverse planning constraints that would reproducibly result in acceptable target and normal tissue dosimetry. RESULTS: We verified the spatial integrity of MRI for treatment planning. MRI anatomic evaluation revealed no significant displacement of the prostate in the left lateral decubitus position, a mean distance of 14.47 mm from the prostatic apex to the penile bulb, and clear demarcation of the neurovascular bundles on postcontrast imaging. Derivation of a class solution for inverse planning constraints resulted in a mean target volume receiving 100% of the prescribed dose of 95.69%, while maintaining a rectal volume receiving 75% of the prescribed dose of <5% (mean 1.36%) and urethral volume receiving 125% of the prescribed dose of <2% (mean 0.54%). CONCLUSION: Systematic evaluation of image spatial integrity, delineation uncertainty, and inverse planning constraints in our procedure reduced uncertainty in planning and treatment. PMID- 15752910 TI - In regard to De Jaeger et al.: significance of plasma transforming growth factor beta levels in radiotherapy for non-small-cell lung cancer (INT J RADIAT ONCOL BIOL PHYS 2004;58:1378-1387). PMID- 15752911 TI - Late effects of radiotherapy for Hodgkin's disease in adolescence. PMID- 15752912 TI - In regard to Pisch et al: placement of 125I implants with the da Vinci robotic system after video-assisted thoracoscopic wedge resection: a feasibility study (INT J RADIAT ONCOL BIOL PHYS 2004;60:928-932). PMID- 15752914 TI - Use of different urethrogram protocols can influence the definition of planning target volume inferior margin in prostate cancer radiotherapy. In regard to Liu et al.: prostate movement during simulation resulting from retrograde urethrogram compared with "natural" prostate movement (INT J RADIAT ONCOL BIOL PHYS 2004;60:470-475). PMID- 15752917 TI - In regard to Boccia et al.: assessment and management of cutaneous reactions with amifostine administration: findings of the ethyol (amifostine) cutaneous treatment advisory panel (ECTAP) (INT J RADIAT ONCOL BIOL PHYS 2004;60:302-309). PMID- 15752919 TI - Management of diabetic dyslipidemia. AB - Identification and management of dyslipidemia is an important element in the multi-factorial approach to prevent coronary heart disease. Diabetic dyslipidemia typically consists of elevated triglyceride, low high-density lipoprotein cholesterol, predominance of small, dense low-density lipoprotein (LDL) particles, and average LDL cholesterol (LDL-C). Lipid-lowering therapy has a beneficial effect on cardiovascular outcomes. Statin treatment is beneficial in patients who are older than 40 years of age, irrespective of the LDL-C value. To achieve lipid targets, attention should be directed first toward nonpharmacologic therapeutic interventions, such as diet, exercise, smoking cessation, weight loss, and improving glycemic control. Although statin therapy is recommended for most subjects, judicious use of combination therapy should be considered in the highest risk subjects. PMID- 15752920 TI - The dyslipidemia of diabetes mellitus: giving triglycerides and high-density lipoprotein cholesterol a higher priority? AB - CVD is the primary cause of morbidity and mortality in patients who have diabetes mellitus. Most such patients have at least one lipid abnormality. Managing these complex lipid disorders is a crucial component of comprehensive diabetes mellitus care and limits the risk for cardiovascular morbidity and mortality. With the high prevalence of mixed lipid disorders, management must focus on all components of the lipid profile. Lowering LDL-C levels remains the first priority, but abnormalities in HDL-C and TG levels also should be treated aggressively. Statins, fibrates, and niacin, along with newer therapies such as ezetimibe, can improve significantly components of the lipid profile. Alone or in combination, these agents can treat the dyslipidemia of diabetes mellitus effectively and safely. PMID- 15752921 TI - Compensatory hyperinsulinemia and the development of an atherogenic lipoprotein profile: the price paid to maintain glucose homeostasis in insulin-resistant individuals. AB - The ability of insulin to stimulate glucose disposal varies sixfold to eightfold among apparently healthy individuals. The only way that insulin-resistant persons can prevent the development of type 2 diabetes is by secreting the increased amount of insulin that is necessary to compensate for the resistance to insulin action. The greater the magnitude of muscle and adipose tissue insulin resistance, the more insulin must be secreted to maintain normal or near-normal glucose tolerance. Although compensatory hyperinsulinemia may prevent the development of fasting hyperglycemia in insulin-resistant individuals, the price paid is the untoward physiologic effects of increased circulating insulin concentrations on tissues that retain normal insulin sensitivity. This article focused on the interplay between insulin resistance at the level of the muscle and adipose tissue and normal hepatic insulin sensitivity; this leads to the atherogenic lipoprotein profile that is characteristic of insulin-resistant individuals. It would be inappropriate to minimize the importance of differential insulin sensitivity in the genesis of the changes in lipoprotein metabolism that increase CVD risk in insulin-resistant persons. It would be equally remiss not to emphasize that differential tissue insulin resistance also is necessary to explain why insulin-resistant/hyperinsulinemic individuals are more likely to develop the clinical syndromes (with the exception of type 2 diabetes mellitus) that are listed in Box 1. PMID- 15752922 TI - Hypertension management in type 2 diabetes mellitus: recommendations of the Joint National Committee VII. AB - HTN in patients who have diabetes should be managed aggressively; the goal BP of less than 130/80 mm Hg should be attained if clinicians seek to reduce cardiovascular morbidity and mortality for these patients. Along with instituting medical therapy after HTN is detected, lifestyle modifications need to be managed aggressively, together with strict glycemic and lipid control. Early management and optimization of treatment of HTN can delay and possibly prevent progression of cardiovascular complications,such as CAD, CKD, peripheral vascular disease, and cerebrovascular disease. Studied approaches to treat HTN in diabetics have included ACEIs and ARBs. Either class of medication, generally in combination with a thiazide diuretic, should be considered as initial therapy. Calcium antagonists, BBs, and alpha-antagonists also have a role in this population of patients, usually as third- and fourth-line add-ons. The importance of using agents that block RAAS is becoming understood better. Typically, three or more antihypertensive medications plus lifestyle interventions are required to achieve a goal BP of less than 130/80 mm Hg. Managing patients who have diabetes and HTN is a dynamic, ever-changing challenge. Early and aggressive antihypertensive therapy pays off;it is hoped that the insights in this article enable clinicians to meet the challenge more successfully. PMID- 15752923 TI - Glycemic management of type 2 diabetes: an emerging strategy with oral agents, insulins, and combinations. AB - The many antihyperglycemic preparations are best used for type 2 diabetes in a logical sequence, using combinations of agents, with clear targets for glycemic control. On the basis of long familiarity, proven benefit and known side effects, and low cost, the sulfonylureas, metformin, and insulin still deserve to be the standard treatments. As shown in the central shaded area of Fig. 4, standard treatment begins with monotherapy and progresses to oral combination therapy and then to two oral agents plus basal insulin. Several triggers for deviation from the standard methods are identified (see Fig. 4). The incidence of each of the conditions that require early individualized treatment has not been studied, but it seems reasonable to estimate no more than 10% each for a strongly symptomatic presentation, inability to use a sulfonylurea or metformin, inability to use insulin, or an early need for prandial therapy. If this estimate is correct, approximately two thirds of patients who are diagnosed with type 2 diabetes should do well with standard therapy for up to 10 years using the standard methods shown. Eventually, many more will need individualized treatment to maintain glycemic control. This scheme is certain to evolve as further information on the nonglycemic benefits (or hazards) of the various therapies appears and as new treatments are released. Notably, agents that mimic or potentiate the effects of gastrointestinal peptides, such as amylin and GLP- 1 analogues and dipeptidyl peptidase IV inhibitors, are likely to alter the current algorithm. For now, systematic application of the scheme (see Fig. 4) should improve the success of treatment greatly from its currently disappointing level. PMID- 15752924 TI - Hospital management of diabetes. AB - The evidence continues to strengthen our understanding that improved glycemic control with the use of insulin therapy may significantly improve morbidity and mortality in hospitalized patients with hyperglycemia, with or without a previous diagnosis of diabetes. However, many questions remain concerning the impact and relative contributions of blood glucose and insulin per se. Nevertheless, the publication of numerous and consistent studies have made it clear that the topic of glycemic management in the hospital requires a larger priority among clinicians caring for these patients. The recently published guidelines by the American Association of Clinical Endocrinologists are the first formal recommendations on this topic,but national guidelines for blood glucose levels cannot take into account all of the different challenges facing different hospitals. This suggests that each institution will require individualization of protocols even though the ultimate metabolic goals are identical. Furthermore, it is not realistic to expect those unfamiliar with diabetes therapy to appreciate all of the nuances and vagaries of insulin treatment. Like any medical treatment, a significant amount of time will need to be invested by the providers involved with the.care of these patients before a mastery of the therapy can be achieved. Nevertheless, because the rewards to our patients can be significant, we need to strive to improve the systems where we work. Individual clinicians with vast experience in diabetes care cannot be successful for the inpatient with diabetes unless the hospital has systems in place to effectively and efficiently facilitate the management of the metabolic needs of this population. The main challenge now is the safe and effective implementation of these guidelines in both small and large hospitals given the limited level of re-sources available in today's medical environment. Therefore, our single most important recommendation is to ensure that all clinicians involved in the management of these patients are in agreement about general philosophies of diabetes management. We would recommend that there are "champions" for each discipline: endocrinology, cardiology, anesthesiology, surgery, nursing,and pharmacy, all of which have developed hospital-specific guidelines for glycemic management. These recommendations can be slowly adapted, one unit at a time, until the entire hospital has transitioned to a more "diabetes-friendly" environment. The ultimate goal of well-controlled glycemia with minimal hypoglycemia should be possible for most hospitals, and we hope this review will assist clinicians in achieving this objective. We await additional outcome research with carefully controlled studies to confirm the value of these recommendations at different levels of glycemic control. We believe that we can already state with confidence that the preliminary evidence shows that, like outpatient diabetes management,metabolic control matters during acute illness. PMID- 15752925 TI - Potential cardiovascular benefits of insulin sensitizers. AB - A multiple risk factor approach is needed in patients who have type 2 diabetes. Because many risk factors are linked with IR, treatment with insulin sensitizers has the potential to modulate these risk factors favorably. TZDs 'have many important effects beyond lowering blood glucose. By targeting IR, they improve many cardiovascular risk factors that are associated with the IR syndrome. In particular, they increase HDL-C, have anti-inflammatory effects, improve endothelial function and fibrinolysis, and decrease carotid intimal thickness; however, no evidence-based studies on cardiovascular outcomes are available to substantiate the potential cardioprotective effects of TZDs. Several clinical trials that were designed to investigate the effect that these agents have on reducing cardiovascular events are well under way. PMID- 15752926 TI - Insulin therapy in people who have dysglycemia and type 2 diabetes mellitus: can it offer both cardiovascular protection and beta-cell preservation? AB - Mounting evidence suggests that insulin therapy may reduce risk for CV events while preserving beta-cell function, and several continuing long-term CV trials are testing these hypotheses explicitly. PMID- 15752928 TI - The prevention of type 2 diabetes mellitus. AB - With a worldwide pandemic of type 2 diabetes upon us, it is imperative that effective and practical preventive strategies be developed for this disease which, once established, carries with it excess morbidity and mortality. This article reviews recently published data on the prevention of type 2 diabetes and compares the demonstrated effectiveness of each strategy, including lifestyle modification and several pharmacologic agents. Although much new information is available, many translational questions remain that chiefly concern the generalizability and applicability of these programs to clinical practice. Recent diabetes prevention guidelines are discussed, and a practical framework for their implementation is presented. PMID- 15752927 TI - Novel pharmacologic agents for type 2 diabetes. AB - After many decades of relative therapeutic stagnation since the initial discovery of insulin, followed by some modifications on its structure and only having sulfonylureas and biguanides for many years, the last decade has seen a surge in new therapeutic options for the management of diabetes. The results of the United Kingdom Prospective Diabetes Study and Kumamoto study indicate the need for aggressive glycemic control and the slow inexorable clinical deterioration associated with type 2 diabetes overtime. The propensity for weight gain and hypoglycemia are the two major limitations that subcutaneous insulin and sulfonylureas have been particularly prone to. The newer antidiabetic medications and those on the horizon attempt to address these limitations. GLP-1 agonists and the DPP-IV inhibitors exploit the innate incretin system to improve glycemia while promoting satiety and weight management. Like GLP-1 related compounds, pramlintide offers the potential to address postprandial hyperglucagonemia associated with type 2 diabetes only limited by the multiple injections and gastrointestinal side effects. The glitazars offer the hope ofa new approach to diabetes care addressing not just glycemia, but dyslipidemia and other components of the metabolic syndrome, though the side effect profile remains a major unknown. The INGAP peptide represents the holy grail of diabetes care as it offers the potential of a new paradigm: that of islet regeneration and potential for a cure. But at this stage, with no human data available, it remains highly speculative. Beyond these and other novel agents being developed to meet the challenge of the worldwide epidemic of diabetes, the central place of insulin in diabetes care cannot be forgotten. In view of this the continued efforts of improvement in insulin delivery, kinetics and action have spurred such innovations as the various inhaled insulins and new insulin analogues. There is cause for guarded optimism and excitement about the years ahead. There is reason to expect that despite the growing burden of diabetes worldwide, we will be better equipped to manage it and its comorbidities and prevent its onset and possibly even cure it. PMID- 15752929 TI - Prevention of cardiovascular outcomes in type 2 diabetes mellitus: trials on the horizon. AB - There has been an explosion of interest in CVD and diabetes mellitus because of the epidemic nature of the diseases and their tight epidemiologic link. Clinical trials over the past decade have built substantial evidence for the role of lipid management, blood pressure control, and antiplatelet therapy in managing risk for CVD in patients who have diabetes mellitus. The many clinical trials underway will not only hone existing recommendations by establishing appropriate targets and techniques for lipid and blood pressure management, but also should demonstrate the role and most appropriate techniques for management of glycemia, flow-limiting coronary lesions, and obesity. PMID- 15752930 TI - New approaches to hypertension management: always reasonable but now necessary. PMID- 15752931 TI - Hypertension as a hemodynamic disease: the role of impedance cardiography in diagnostic, prognostic, and therapeutic decision making. AB - Hypertension is the most common cardiovascular disease, affecting approximately 60 million Americans. Despite the importance of this condition, only the minority of patients are appropriately identified and treated to reach recommended blood pressure (BP) goals. Although historically defined as an elevation of BP alone, hypertension is characterized by abnormalities of cardiac output, systemic vascular resistance, and arterial compliance. These hemodynamic aspects of hypertension have implications for diagnosis, risk stratification, and treatment. Impedance cardiography (ICG) has emerged as a unique and highly accurate noninvasive tool that is used to assess hemodynamic parameters. Measurement of the various hemodynamic components using ICG in those with hypertension allows more complete characterization of the condition, a greater ability to identify those at highest risk, and allows more effectively targeted drug management. This article reviews the importance of hemodynamic factors in hypertension and the evolving role of ICG technology in the assessment and management of this important cardiovascular condition. PMID- 15752932 TI - Reproducibility of impedance cardiography parameters in outpatients with clinically stable coronary artery disease. AB - BACKGROUND: Impedance cardiography (ICG) is a noninvasive method of determining hemodynamic parameters. It is clinically important to determine whether any change in ICG parameters occur due to changes in disease status or therapeutic interventions, or due to normal hemodynamic and technology variability. The objective of this study was to establish the intra- and inter-day reproducibility of ICG in a stable population with coronary artery disease (CAD). METHODS: A prospective, time series design was used. The study group consisted of 96 clinically stable CAD patients in an outpatient cardiac rehabilitation program. Measurements of ICG hemodynamic parameters were obtained at four points in time: after 5 and 10 min of rest on the first day and after 5 and 10 min of rest on a second day, 1 week later. RESULTS: There were small but significant intra-day changes between the 5- and 10-min hemodynamic measures. Mean absolute percent changes in intra-day hemodynamic measures were <8%. High intra-day correlation was observed, ranging from 0.85 for mean arterial pressure to 0.99 for thoracic fluid content. There were expectedly larger inter-day hemodynamic variation of up to 18% and lower inter-day correlation for all ICG parameters ranging from 0.66 to 0.88. CONCLUSIONS: Impedance cardiography measurements demonstrate both intra- and inter-day reproducibility within clinically acceptable ranges in a clinically stable population of CAD patients. The expected ranges of variation can be used to gauge whether a patient's hemodynamic status has changed because of disease or intervention. PMID- 15752933 TI - Noninvasive hemodynamic profiles in hypertensive subjects. AB - BACKGROUND: Hypertension is a disease state characterized by increased blood pressure (BP) associated with hemodynamic abnormalities, including elevated systemic vascular resistance index (SVRI); and altered cardiac index (CI). The objective of this study was to use noninvasive impedance cardiography (ICG) to evaluate hemodynamic characteristics of subjects with and without hypertension. METHODS: A total of 19 healthy nonhypertensive and 136 hypertensive subjects were retrospectively evaluated. Hemodynamic parameters were measured with ICG and included CI, SVRI, total arterial compliance index (TACI), and thoracic fluid content (TFC); these were compared with subject type, blood pressure value, demographics, and medications. RESULTS: The BP levels of healthy and hypertensive subjects were 117/71 and 154/90 mm Hg, respectively (P < .0001). Subjects with prehypertension had a lower TACI (0.97 v 1.21, P < .05) compared with those with a normal BP, ie, <120/80 mm Hg. Hypertensive subjects had significantly lower SI, CI, TACI, and TFC and significantly higher SVRI. Subjects with stage 2 hypertension had higher SVRI (4149 v 3418 dyne.sec(2).cm(-5).m(2), P < .01) and lower TACI (0.61 v 0.53 mm Hg/mL/m(2), P < .05) than those with stage 1 hypertension. Compared with subjects with controlled hypertension, normal subjects had significantly lower SVRI (1996 v 2746 dyne.sec(2).cm(-5).m(2), P < .0001) and significantly higher CI (3.23 v 2.63 L/min/m(2), P < .001), SI (48.2 v 37.4 mL/m(2), P < .0001), TACI (1.08 v 0.85 mm Hg/mL/m(2), P < .01), and TFC (29.1 v 24.1/kOhm, P < .0001). The parameters of TACI, SVRI, and CI demonstrated modest correlation (-0.75, 0.62, and -0.30), respectively, with SBP. In the 54 subjects with BP <140/90 mm Hg, SVRI values varied significantly, with 32 subjects (39.2%) with SVRI values in the high range (>2483 dyne.sec(2).cm( 5).m(2)). CONCLUSIONS: Hemodynamic parameters from ICG displayed significantly different hemodynamic profiles between hypertensive and nonhypertensive subjects. However, significant individual variation of hemodynamic status exists. Hemodynamic measurements with ICG characterize hemodynamic status and may be helpful in diagnostic, prognostic, and therapeutic decision making in hypertensive subjects. PMID- 15752934 TI - Noninvasive hemodynamic assessment of the effect of mean arterial pressure on the amplitude of pulse pressure. AB - BACKGROUND: The present study compares the relationships between mean arterial pressure (MAP) and pulse pressure (PP) in young and older men and whether MAP determines age-related changes in PP. METHODS: Impedance cardiography was used to evaluate systemic hemodynamics noninvasively in 189 unmedicated men referred to our hypertension unit. Patients were divided by age according to the median value (< and >/=40 years). RESULTS: In younger patients, increasing supine MAP was associated with a transition in the blood pressure pattern from systolic to diastolic, whereas in older patients, increasing MAP was associated with change from diastolic to systolic hypertension. In young patients elevation in MAP was associated with a parallel decrease in PP (P < .001) and stroke volume index (SVI) (P < .001), whereas in older patients higher MAP was associated with higher PP (P < .001) and PP/SVI ratio (P < .001), a measure of arterial stiffness. When the sample was divided according to the median value of MAP (< and >/=101 mm Hg), differences in the age-related change in PP and SVI became apparent. With a lower MAP, changes in PP and SVI remained parallel until the sixth decade, after which they began a progressive dissociation. In contrast, at higher MAP, the dissociation between PP and SVI began two decades earlier, indicating an acceleration of the age-related increase in arterial stiffness. Therefore, the relationship between MAP and PP was negative in younger men at the expense of a decrease in SVI, and positive in older men due to increasing arterial stiffness. CONCLUSIONS: Noninvasive hemodynamic parameters from impedance cardiography provide a useful method to characterize the mechanism of increased blood pressure. PMID- 15752935 TI - Prognostic value of hemodynamic findings from impedance cardiography in hypertensive stroke. AB - BACKGROUND: Impedance cardiography (ICG), a noninvasive method of hemodynamic monitoring, is a useful tool in the care of critically ill patients. Use of this technology shows promise in the hemodynamic assessment of hypertensive stroke patients. This study describes the different ICG findings of patients with hypertensive stroke and correlates parameters with patient outcome. METHODS: Adult patients with stroke admitted to the intensive care unit (ICU) who were on antihypertensive medications or had hypertensive blood pressure (BP) levels at the time of admission were included. Patients were classified as having ischemic or hemorrhagic stroke on the basis of imaging studies. The ICG was done in the initial 24 h of admission using the BioZ Cardiac Output Monitor (CardioDynamics, Inc., San Diego, CA) and 18 hemodynamic parameters were analyzed. Statistical analysis was performed to assess correlation of hemodynamic findings with the type of stroke and survival to hospital discharge. RESULTS: Fifty-two adult patients with a mean age of 60.5 +/- 12.4 years were included. Of this population, 29 had ischemic stroke and 23 had hemorrhagic stroke. Seventeen patients died during hospital stay. Overall, noninvasive hemodynamic parameters of stroke patients showed a high systemic vascular resistance index (SVRI) and systemic vascular resistance (SVR), and decreased cardiac output (CO), stroke index (SI), and stroke volume (SV). Among the ischemic stroke group, nonsurvivors had a significantly higher mean SVRI and mean SVR compared to the survivors. In contrast, among the hemorrhagic stroke patients, nonsurvivors had a significantly lower mean SVRI and SVR compared to survivors. Except for SI and BP, there was no difference in the noninvasive hemodynamic parameters between the ischemic stroke group and the hemorrhagic stroke group. CONCLUSIONS: In this population of hypertensive stroke patients admitted to the ICU, ICG showed an elevated SVRI and SVR and depressed CO, SV, and SI. In the ischemic stroke group, higher SVR and SVRI were associated with in-hospital death, whereas in the hemorrhagic stroke group, lower SVR and SVRI were associated with in-hospital death. The ICG may provide significant prognostic information in patients admitted with hypertensive stroke. PMID- 15752936 TI - Diagnostic ability of B-type natriuretic peptide and impedance cardiography: testing to identify left ventricular dysfunction in hypertensive patients. AB - BACKGROUND: Patients with hypertension are at high risk for the development of left ventricular dysfunction (LVD). Echocardiography is considered to be the gold standard for diagnosis of LVD; but its cost, complexity, and availability prevents its use for frequent evaluation. Brain natriuretic peptide (BNP) and N terminal BNP (NT-BNP) can identify heart failure in dyspneic patients. Impedance cardiography (ICG) is a noninvasive method of measuring hemodynamic and electromechanical timing parameters. The objective of this study was to determine the ability of BNP, NT-BNP, and ICG to detect the presence of LVD in patients with hypertension. METHODS: A convenience sample of subjects undergoing echocardiography who had a history of hypertension or current systolic blood pressure >/=140 mm Hg were enrolled and retrospectively evaluated. Patients with known LVD were excluded. Diagnosis of LVD was determined by the presence of systolic or diastolic dysfunction, valvular or wall motion abnormalities, or left ventricular hypertrophy. RESULTS: A total of 193 subjects were enrolled: 189 men and four women, age 68.8 +/- 11.7 years. Multivariate regression analysis of history and symptoms, BNP, and ICG parameters identified significant predictor variables for LVD including cardiac index (P = .005), left cardiac work index (P = .008), BNP (P = .017), arrhythmia (P = .023), angina (P = .034), and systemic vascular resistance (P = .048). Receiver operating characteristic (ROC) analysis determined the area under the ROC curve (AUC) of BNP (0.60), NT-BNP (0.67), ICG velocity index (0.66), composite ICG (0.66), ICG combined with BNP (0.70), and ICG combined with NT-BNP (0.73). CONCLUSIONS: In this high-risk hypertensive population, BNP, NT-BNP, and ICG were useful to identify the presence of LVD. The use of ICG with natriuretic peptide testing may improve the ability to detect LVD. PMID- 15752937 TI - Systolic blood pressure does not reliably identify vasoactive status in chronic heart failure. AB - BACKGROUND: In chronic heart failure (CHF), titration of vasodilating medications is often guided by monitoring of systolic blood pressure (BP). However, systolic BP may not indicate the patient's true vasoactive status-best approximated by systemic vascular resistance-because cardiac output is also a contributing factor. Impedance cardiography (ICG) is a validated noninvasive method of measuring cardiac output and systemic vascular resistance. METHODS: To evaluate the relationship between systolic BP and systemic vascular resistance index (SVRI) in patients with CHF, we retrospectively evaluated the systolic BP and SVRI from 71 consecutive patients during 615 CHF clinic visits. Measurement of systolic BP was through the oscillometric method and SVRI through ICG (BioZ ICG Monitor, CardioDynamics, San Diego, CA). Absolute values and relative changes in systolic BP and SVRI were compared and characterized by systolic BP grouping. RESULTS: The 71 patients were an average of 69.3 +/- 12.2 years, New York Heart Association functional class 2.52 +/- 0.6, 46.5% men, and 47.8% ischemic etiology. Frequency by systolic BP grouping was: <100 mm Hg in 67 subjects (10.9%), 100 to 119 mm Hg in 245 subjects (39.8%), and >/=120 mm Hg in 303 subjects (49.3%). The correlation (R(2) value) of systolic BP to SVRI was 0.21 (N = 615), and change in systolic BP to change in SVRI from previous visit was 0.27 (N = 547). In 138 visits in which systolic BP did not change by 5 mm Hg or more, SVRI changed by 20% or more in 41 (29.7%). In the 67 visits in which systolic BP was below 100, only 6 (9.0%) had low SVRI and 57 (85.1%) had normal SVRI. In the 245 visits with systolic BP 100 to 119, 58 (23.7%) had high SVRI. CONCLUSIONS: Measurement of systolic BP alone does not reliably indicate the degree of vasoconstriction or vasodilation that exists in patients with CHF. Measurement of SVRI by ICG may help guide determination of need and tolerance for vasodilating medications in CHF. PMID- 15752938 TI - Use of noninvasive hemodynamics in hypertension management. AB - Hypertension is a chronic disease that is controlled in the United States in only 34% of those taking antihypertensive medications. Because hypertension is a hemodynamic disorder, the patient's hemodynamic profile before and after medical intervention may assist in the decision and evaluation of ongoing antihypertensive therapy. There are several medication classes used in the management of hypertension and it is challenging at best for the clinician to determine the optimal therapeutic combination of medications for each patient. Physician perceptions and patient symptoms are examples of barriers affecting the management and control of hypertension. Impedance cardiography is a noninvasive monitoring technique that provides reliable and reproducible hemodynamic measurements. Three case studies are presented that illustrate how hemodynamic parameters were used to achieve hypertension control in the outpatient setting. PMID- 15752939 TI - Time for focus on morning hypertension: pitfall of current antihypertensive medication. PMID- 15752940 TI - Relationship among end-stage renal disease, hypertension, and sleep apnea in nondiabetic dialysis patients. AB - BACKGROUND: Cardiovascular diseases (CVD) are responsible for more than 50% of the deaths in patients with end-stage renal disease (ESRD). Sleep apnea (SA) has been recognized as a risk factor for CVD. Previous studies have shown a higher prevalence of SA among patients on dialysis. METHODS: Forty-five nondiabetics patients with ESRD underwent a polysomnographic analysis with concomitant clinical evaluation and laboratory tests. Fourteen patients (31.1%) presented with an apnea/hypopnea index (AHI) more than 5, confirming a high prevalence of SA. We observed abnormal sleep pattern with high percentages of sleep stage 1 and low percentages of sleep stages 3 and 4. RESULTS: Patients with AHI more than 5 presented higher levels of systolic, diastolic, and mean blood pressures (MBP) as compared with those with AHI less than 5 (P < .05). When other variables were compared (age, time of dialytic treatment, cause of ESRD, use of antihypertensive drugs, body mass index, serum levels of hemoglobin, hematocrit, creatinine, KT/V index, pH, bicarbonate, parathormone, and alkaline phosphatase), no differences were found between the two groups. In a logistic regression model, MBP and age more than 40 years were positively related to the presence of SA. CONCLUSIONS: Our study is in agreement with previous works and shows that patients with ESRD have a higher SA index compared to those with normal renal function. In spite of having higher levels of BP no other parameter was different among apneic and nonapneic patients. Hypertension may play a pivotal role linking SA and CVD. PMID- 15752941 TI - Inhibition of brain angiotensin-converting enzyme by peripheral administration of trandolapril versus lisinopril in Wistar rats. AB - BACKGROUND: Peripheral administration of blockers of the renin-angiotensin system (RAS) may affect the RAS in the brain to a variable degree. In the present study, we determined inhibition of angiotensin-converting enzyme (ACE) in the brain after peripheral administration of a lipophilic (trandolapril) versus hydrophilic (lisinopril) ACE inhibitor. METHODS: Trandolapril (0.2, 1, and 5 mg/kg/day, subcutaneously) was compared with lisinopril (2, 10, and 50 mg/kg/day, subcutaneously), each for 6 days. At 4 and 24 h after the last dose, (125)I-351A binding on brain ACE was measured. RESULTS: Trandolapril and lisinopril caused similar inhibition of ligand binding outside the blood-brain barrier (BBB). However, inside the BBB, trandolapril was more effective at low and medium doses (for lisinopril, 28% to 51% inhibition at a dose of 2 mg, 63% to 72% at 10 mg, and 84% to 86% at 50 mg; and for trandolapril, 62% to 68% inhibition at a dose of 0.2 mg, 84% to 87% at 1 mg, and 88% to 93% at 5 mg). In contrast, in the brain structures caudate putamen and globus pallidus, lisinopril inhibited ligand binding better than trandolapril (for lisinopril 30% to 44% at a dose of 2 mg and 71% to 74% at 10 mg, versus for trandolapril 21% to 27% at 0.2 mg and 51% to 63% at 1 mg). At 24 h after the last dose, inhibition by trandolapril persisted more than inhibition by lisinopril both outside and inside the BBB. CONCLUSIONS: These results suggest that peripheral administration of even hydrophilic ACE inhibitors can result in marked inhibition of brain ACE inside the BBB but that different brain structures show variable inhibition. PMID- 15752942 TI - In-hospital complications of acute myocardial infarction in hypertensive subjects. AB - BACKGROUND: Recent studies have shown a worse in-hospital outcome in hypertensive than in normotensive patients with acute myocardial infarction (AMI), which has been attributed to more frequent complications. The aim of this study was to investigate clinical patterns, risk factors, and in-hospital complications in hypertensive and normotensive patients with AMI. METHODS: Of 4994 consecutive patients with AMI admitted to the intensive care unit, hypertensive patients with first infarction (n = 915; mean age 68.8 +/- 11.4 years) and 915 gender- and age matched normotensive subjects were retrospectively studied. RESULTS: In the univariate analysis, hypertensive subjects presented more frequently non-Q-wave infarction and ST segment depression than did normotensive subjects, even if hypertensive subjects more frequently had diabetes, dyslipidemia, renal failure, peripheral artery disease, cerebrovascular disease, and chronic obstructive pulmonary disease (P < .01 for all). Hypertensive subjects less frequently presented with cardiogenic shock (4.0% v 11.6%; P < .01), atrioventricular block (4.9% v 7.4%; P = .02), ventricular fibrillation (2.2% v 3.7%; P = .04), cardiac rupture (0.1% v 0.9%; P = .02), and ventricular thrombosis (0.5% v 1.5%; P < .03), and a higher frequency of paroxysmal atrial fibrillation (9.2 v 5.6%; P < .01). Mortality was significantly higher in patients with anterior versus inferior infarction, for all normotensive and hypertensive subjects (13.7% v 7.1%; P < .001), but mortality was remarkably higher in normotensive than in hypertensive subjects (17.8% v 6.2%; P < .001), regardless of infarction site (anterior, 11.2% v 4.1%; P < .001; inferior, 4.4% v 1.9%; P < .001). CONCLUSIONS: Hypertensive subjects with first AMI have a better in-hospital outcome than age- and gender-matched normotensive subjects, perhaps due to a less severe extension of the infarction area or to a different pathophysiologic mechanism. PMID- 15752943 TI - Left ventricular mass and mechanics in mild-to-moderate hypertension: effect of nebivolol versus telmisartan. AB - BACKGROUND: The aim of this study was to compare the effects of nebivolol and telmisartan on left ventricular mass (LVM) and midwall mechanics in mild-to moderate hypertension. METHODS: A total of 40 patients with mild-to-moderate hypertension were randomized to receive either nebivolol (2.5 to 5.0 mg/day) or telmisartan (40 to 80 mg/day) to achieve a target diastolic blood pressure of <90 mm Hg. Blood pressure (BP) was measured with sphygmomanometry, and LVM and midwall fractional shortening (mFS) were estimated by two-dimensionally guided M mode echocardiography at baseline and at 3-month follow-up. RESULTS: Age, sex distribution, and baseline SBP and DBP and heart rate were similar in the two groups. Both nebivolol and telmisartan reduced systolic (156 +/- 7 v 124 +/- 8 mm Hg, P < .01, and 153 +/- 5 v 120 +/- 7 mm Hg, P < .01, respectively) and DBP (99 +/- 4 v 80 +/- 2 mm Hg, P < .01 and 98 +/- 2 v 80 +/-2 mm Hg, P < .01, respectively) and increased mFS (16% +/- 2% v 19% +/- 2%, P < .01, and 15% +/- 2% v 18% +/- 2%, P < .01, respectively). The LVM indices decreased significantly with both nebivolol (98 +/- 16 v 84 +/- 13 g/m(2), P < .01) and telmisartan (97 +/- 13 v 83 +/- 8 g/m(2), P < .01). We found that mFS was inversely related to DBP in the nebivolol but not in the telmisartan group. CONCLUSIONS: In mild-to moderate hypertension, nebivolol and telmisartan are equally effective in reducing BP and increasing mFS. There may be differences between nebivolol and telmisartan regarding the mechanism of increase in mFS. PMID- 15752945 TI - A double-blind, dose-response study of losartan in hypertensive children. AB - BACKGROUND: The aim of this study was to determine the dose-response relationship for losartan, 2.5 to 100 mg, and to assess the safety and tolerability of losartan in hypertensive children 6 to 16 years of age. METHODS: This was a multicenter, randomized, double-blind, dose-response study. In Period 1, a total of 175 patients were stratified by weight (<50 kg and >/=50 kg) and randomized to one of three dose groups by stratum (low, 2.5/5.0 mg; middle, 25/50 mg; or high, 50/100 mg) for 3 weeks. The ratio of the three dose levels for both weight strata was 1:10:20. In Period 2, patients in each dose group were randomized to continue the same treatment or placebo washout for 2 additional weeks. RESULTS: In Period 1, sitting trough diastolic blood pressure (DBP) decreased in a dose-dependent manner (P < .0001). At week 3, changes in DBP from baseline in the low-, middle-, and high-dose groups were -6.0 mm Hg, -11.7 mm Hg, and -12.2 mm Hg, respectively. In Period 2, DBP increased significantly in patients who switched from middle- and high-dose losartan to placebo (mean increase 6.0 mm Hg, P = .003) relative to DBP in patients who remained on active treatment; however, these levels remained stable in those patients who switched from low-dose losartan to placebo (mean increase 1.1 mm Hg, P = .628). CONCLUSIONS: In hypertensive children 6 to 16 years of age, losartan given once daily reduced blood pressure in a dose dependent fashion. A once-daily starting dose of losartan, 0.75 mg/kg (maximum 50 mg) effectively lowered DBP within 3 weeks. Losartan up to a dosage of 1.44 mg/kg (maximum 100 mg) once daily is generally well tolerated. PMID- 15752944 TI - Insulin sensitivity and endothelial function in hypertension: a comparison of temocapril and candesartan. AB - BACKGROUND: Recent studies have suggested that angiotensin-converting enzyme inhibitors (ACEi) have a more pronounced effect on endothelial function (END) than angiotensin II receptor blocker (ARB); however, whether this pronounced effect is more beneficial to patients with insulin sensitivity (IS) remains uncertain. The present study compared the effects of ACEi and ARB on END and IS in patients with hypertension. METHODS: A total of 23 patients with hypertension were given either ACEi or ARB alternatively in a cross-over manner for 8-week intervals. Both END and IS were examined after each treatment period; END was assessed by the response of forearm blood flow to reactive hyperemia and IS by an insulin tolerance test. The plasma levels of bradykinin (BK), NOx, tumor necrosis factor (TNF-alpha), and adiponectin (Adi) were also measured after each treatment. RESULTS: We found that END, BK, and NOx were higher after the ACEi treatment than after the ARB treatment. Although the IS and the Adi levels were similar after both treatments, the TNF-alpha level was lower after the ARB treatment than after the ACEi. CONCLUSIONS: We conclude that ACEi and ARB may have similar effects on insulin sensitivity, irrespective of the more pronounced effects of ACEi on endothelial function. The BK-NO pathway might contribute, at least in part, to the pronounced effect of ACEi. On the other hand, the underlying mechanisms affecting insulin sensitivity might differ for both treatments. These results suggest that endothelial function is not a major determinant of insulin sensitivity under physiologic conditions. PMID- 15752947 TI - Average blood pressure and cardiovascular disease-related mortality in middle aged women. AB - BACKGROUND: The aim of this study was to assess which average blood pressure (BP) component (ie, systolic BP [SBP], diastolic BP [DBP], pulse pressure [PP], or mean arterial pressure [MAP]), is most strongly related to cardiovascular disease (CVD)-related mortality and to evaluate whether the strength of the relation varies with follow-up time. METHODS: This was a prospective cohort study. The studied cohort comprised a population of postmenopausal women (n = 7813) between the ages of 49 and 66 years of age, of whom four BP measurements were available, obtained at four different time points. Average BP, ie, the mean of the four measurements divided by the standard deviation, was entered in Cox proportional hazards models to facilitate direct comparison. Hazard ratios (HR) were calculated adjusted for age, body mass index, presence of diabetes mellitus, smoking habit, and use of BP-lowering medication. In addition analyses were repeated in strata of follow-up time (10, 15, and 20 years). RESULTS: During a mean follow-up of 13.1 years, 463 CVD-related deaths occurred. For SBP and MAP the highest HR for CVD mortality were found; however, the confidence intervals (CI) overlapped (SBP: HR = 1.43, 95% CI = 1.30 to 1.58; DBP: HR = 1.35, 95% CI = 1.23 to 1.50; PP: HR = 1.30, 95% CI = 1.19 to 1.42; MAP: HR = 1.43, 95% CI = 1.30 to 1.58). Analyses in strata of follow-up time did not show a difference in strength of the associations with increasing follow-up time. CONCLUSIONS: In this prospective follow-up study of postmenopausal women, SBP and MAP seemed to be strongest related with CVD-related death; however the CI of the HR overlapped. PMID- 15752946 TI - Normalization for body size and population-attributable risk of left ventricular hypertrophy: the Strong Heart Study. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is identified by left ventricular mass (LVM) normalized by body surface area (BSA) or height (in meters) also raised to allometric powers. The presence of LVH detected by these indices predicts increased cardiovascular (CV) events. Whether different indexations of LVH differ in their ability to predict excess risk is unknown. METHODS: A total of 2400 subjects, (1589 women and 811 men), 59 +/- 8 years of age and without prevalent CV disease, valve disease or wall motion abnormalities and high prevalence of obesity were followed for an average of 86 months. Reference values (mean +/- 1.96 SD) for LVM/BSA, LVM/BSA(1.5), LVM/m, LVM/m(2.7), and LVM/m(2.13) were obtained in 251 normal participants and population-attributable risk percent (PAR%) for fatal and nonfatal CV events were calculated from prevalence of LVH and hazard ratios (HR). RESULTS: In the entire population or in hypertensive participants, prevalence of LVH was higher for LVM/m(2.7) (20% and 28%) and LVM/m(2.13) (18% and 25%) than for BSA (7% and 11%). Age and sex-adjusted PAR% for LVM/m(2.7) or LVM/m(2.13) were on average 1.8-fold greater than for LVM/BSA in the entire population, and 1.6-fold greater in hypertensive participants, differences that were statistically significant. CONCLUSIONS: The presence of LVH identified by LVM normalized for height to allometric powers is associated with a higher proportion of incident CV events than is LVH detected by normalization for BSA and is convenient for identification of individuals at high risk and in need of preventive intervention in populations with high prevalence of obesity. Allometric power methods allow detection of prognostically adverse, obesity related LVH, which is unidentified using BSA. PMID- 15752948 TI - Heart failure after long-term supravalvular aortic constriction in rats. AB - BACKGROUND: Pressure overload in humans follows a chronic and progressive course, often resulting in eventual cardiac decompensation and death. Animal models of heart failure generally fail to mimic the temporal features observed in human disease often covering a major portion of the life span, and findings of short term studies are of uncertain applicability. The purpose was to determine whether chronic pressure overload introduced gradually in young normotensive rats would lead predictably to heart failure and to characterize specific phenotype features that have been well documented in another model of heart failure. METHODS: Rats underwent banding of the ascending aorta at 7 weeks of age such that the hemodynamic load increased gradually with ontogenic growth. Two groups of hypertrophied hearts from aortic-banded rats, with and without signs of heart failure, were compared with those of control rats at a mean age of 11 months. RESULTS: Hearts of aorta-banded rats underwent a transition from stable compensated hypertrophy to heart failure that was characterized by augmented hypertrophy, depressed contractile function, elevated fibrosis, increased myocardial stiffness, and marked alterations in the expression of genes encoding contractile, regulatory, and extracellular matrix proteins. CONCLUSIONS: Gradual constriction of the rat aorta resulted in heart failure after a variable length of time (3 to 18 months). Despite differences in genotype, the ultimate phenotype associated with the transition to failure in the aorta-banded rat is nearly identical to that observed in the aged spontaneously hypertensive rat (SHR), with a few notable differences. The findings suggest that a common heart failure phenotype follows long-term pressure overload regardless of the underlying etiology. PMID- 15752949 TI - Dietary fish oil prevents vascular dysfunction and oxidative stress in hyperinsulinemic rats. AB - BACKGROUND: Fish oil has been shown to improve blood pressure (BP) in some disease states by an unknown mechanism. We tested the ability of fish oil to prevent vascular dysfunction in fructose-fed rats, a model of insulin resistance and hypertension. METHODS: Rats were placed on three diets: 1) regular rat diet (control); 2) diet containing 60% fructose (FFR); or 3) diet containing 60% fructose and 4.4% fish oil (FFR+FO). After 8 weeks, blood, heart, aorta, and mesenteric artery tissue were collected from each animal. Secondary branch segments of mesenteric arteries were isolated for vascular reactivity studies. RESULTS: Systolic BP increased significantly in the FFR but was reduced to control levels by the addition of fish oil to the diet. In the mesenteric artery segments from FFR, the dose-response curves to acetylcholine were significantly shifted to the right compared with those of control rats and rats on the fish oil diet. Expression of endothelial nitric oxide synthase (eNOS) protein and mRNA was reduced in the FFR aortas and hearts, and this reduction was reversed by the fish oil. Dietary fish oil prevented the hyperlipidemia that occurred in the FFR but did not prevent hyperinsulinemia. Plasma concentrations of hydrogen peroxide, 8 isoprostane, and monocyte chemoattractant protein-1 were significantly elevated in the FFR and were significantly lower in the FFR treated with fish oil. CONCLUSIONS: These results demonstrate that dietary fish oil prevents vascular dysfunction in FFR and that this effect of fish oil is associated with increased eNOS expression and decreased oxidative stress. PMID- 15752951 TI - The effect of rosiglitazone on urine albumin excretion in patients with type 2 diabetes mellitus and hypertension. AB - BACKGROUND: Thiazolidinediones are antidiabetic agents that improve insulin sensitivity (IS). Accumulating data indicate that these agents provide beneficial effects beyond glycemic control, such as improvement in vascular function. The aim of this study was to determine the effect of rosiglitazone on urine albumin excretion (UAE) in patients with type 2 diabetes mellitus (DM) and hypertension. METHODS: The study involved 20 subjects with type 2 DM who were already on 15 mg glibenclamide daily but were achieving poor glycemic control and who had either poorly controlled or newly diagnosed hypertension. In these patients, rosiglitazone (4 mg daily) was added to the existing therapeutic regimen for 26 weeks. At baseline and the end of the treatment, subjects gave a 24-h urine collection for direct measurement of albumin and a spot specimen for determination of the albumin-to-creatinine ratio (ACR). Subjects also had a hyperinsulinemic euglycemic clamp and an ambulatory blood pressure (BP) monitoring. RESULTS: At the end of the study, UAE was significantly reduced versus baseline, as measured either directly in the 24-h collection (22.4 +/- 4.6 v 13.8 +/- 3.0 mg/day, P < .05) or with ACR (20.9 +/- 3.8 v 14.0 +/- 2.8 mg/g, P < .05). The percentage changes in UAE (DeltaALB for the 24-h collection and DeltaACR for ACR) correlated with the respective changes in IS (r = -0.64, P < .01 for DeltaALB and r = -0.48, P < .05 for DeltaACR), systolic BP (r = 0.63, P < .01 and r = 0.58, P < .01 respectively), and diastolic BP (r = 0.56, P < .05 and r = 0.50, P < .05 respectively). CONCLUSIONS: In this study, treatment of type 2 diabetic hypertensive patients with rosiglitazone significantly decreased UAE. Lowering of BP and improvement of IS should play roles in this UAE reduction. PMID- 15752952 TI - Acute hyperinsulinemia reduces plasma leptin levels in insulin-sensitive Japanese men. AB - BACKGROUND: Elevated plasma leptin levels have been demonstrated in obesity and hypertension. These conditions are documented as insulin-resistant states with sympathetic overactivity. However, the relation between plasma insulin, leptin levels, and sympathetic nerve activity, especially after meals, has not been established. METHODS: To evaluate the impact of insulin sensitivity and obesity on the response of plasma leptin to acute physiologic insulin elevation, we studied 31 nonobese (body mass index [BMI] < 25 kg/m(2)) and 38 overweight or obese (obese; BMI >/= 25 kg/m(2)) normotensive men. Using the Homeostasis Model Assessment of Insulin Resistance (HOMA-IR), the subjects were subdivided into insulin-sensitive (HOMA-IR <2.5) and insulin-resistant (HOMA-IR >/=2.5) groups. There were 11 nonobese and 28 obese men who were insulin-resistant. Blood pressure (BP), plasma glucose, insulin, leptin, and norepinephrine (NE) levels were measured before and after 75-g oral glucose loading every 30 min for 120 min. RESULTS: In nonobese subjects, fasting plasma insulin and leptin levels and areas-under-the-curves (AUC) for insulin and leptin were significantly lower in the insulin-sensitive group than in the insulin-resistant group. In the insulin sensitive group regardless of BMI, plasma leptin levels decreased after glucose loading. Plasma glucose, insulin, NE, and BP levels increased in nonobese insulin sensitive subjects after glucose loading, whereas in obese insulin-sensitive subjects, plasma NE and BP did not change in response to glucose. In insulin resistant subjects regardless of BMI, marked elevations in plasma insulin did not cause any change in plasma leptin, NE, and BP levels. CONCLUSION: These results demonstrate that insulin sensitivity and adiposity affect the response of leptin and sympathetic nerve activity to acute postprandial hyperinsulinemia. PMID- 15752950 TI - Short-term oral folic acid supplementation enhances endothelial function in patients with type 2 diabetes. AB - BACKGROUND: Endothelial dysfunction and arterial stiffening are commonly observed in type 2 diabetes. These abnormalities might be secondary to increased plasma concentrations of homocysteine. We sought to determine whether oral folic acid supplementation, by lowering homocysteine levels, enhanced endothelial function and reduced arterial stiffness in type 2 diabetes. METHODS: Twenty-six type 2 diabetic patients (age 56.5 +/- 0.9 years, diabetes duration 5.5 +/- 0.6 years, means +/- SEM) with no history of cardiovascular disease received 5 mg/d of oral folic acid or placebo for 4 weeks in a double-blind, randomized controlled, parallel group trial. The following parameters were measured before and after treatment: 1) endothelial function (forearm arterial blood flow during local intra-arterial administration of endothelium-dependent [acetylcholine 1.5, 4.5, and 15 microg/min] and endothelium-independent [sodium nitroprusside 1, 2, and 4 microg/min] vasodilators); and 2) carotid-radial and carotid-femoral pulse wave velocity. RESULTS: Folic acid reduced plasma homocysteine concentrations and enhanced endothelium-dependent vasodilatation during each acetylcholine infusion rate (mean and 95% confidence interval post versus pretreatment differences in forearm arterial blood flow ratio between the infused and control arm +0.19 (0.03 0.35), P < .01; +0.39 (0.02-0.81), P < .05; and +0.40 (0.09-0.89), P < .05, respectively). Endothelium-independent vasodilatation and pulse wave velocity were not affected. No significant changes in forearm arterial blood flow and pulse wave velocity were observed in the placebo group. Multiple regression analysis showed that changes in folic acid, but not homocysteine, concentrations independently described changes in maximal endothelium-dependent vasodilatation. CONCLUSIONS: Short-term oral folic acid supplementation significantly enhances endothelial function in type 2 diabetic patients, independent of homocysteine lowering. PMID- 15752953 TI - What is the accuracy of clinic blood pressure measurement? AB - BACKGROUND: In clinical practice, blood pressure (BP) is frequently measured at the end of the visit in patients sitting on one side of the bed and not on a chair according to guidelines. METHODS: In 540 consecutive subjects with essential hypertension (EH) attending a hospital outpatient clinic, BP was measured in the following sequence: 1) patient seated on chair for at least 5 min, 2) patient supine, 3) patient seated on bed, and 4) patient standing for a few minutes. RESULTS: We found that mean (+/-SEM) BP was 143.5/87.2 +/- 0.9/0.5, 153.4/89.7 +/- 1.0/0.5, 148.9/90.9 +/- 1.0/0.5, and 144.8/91.7 +/- 1.0/0.6 mm Hg, respectively (P < .05 v position 1 for all). In 14% of patients, either systolic BP (SBP) or diastolic BP (DBP) was above the conventional upper limits of normality in the seated-on-bed but not in the recommended seated-on-chair position ("false" high clinic BP), whereas SBP and DBP were "false" normal (below limit for bed-seated and above limit for chair-seated position) in only 6% and 2% of patients, respectively. Overall, SBP and DBP increments from the chair- to the bed-seated position were inversely related to the baseline chair-seated values; systolic increments were directly related to age, in particular in the subgroup of untreated EH (n = 70), and to body mass index. A gender-related difference was apparent, as female subjects had more pronounced increments in SBP (+7.4 +/- 0.8 v +3.5 +/- 0.7 mm Hg) and DBP (+4.4 +/- 0.5 v 2.9 +/- 0.4 mm Hg) than did male subjects (P < .05 for both). CONCLUSIONS: Clinic SBP and DBP are overestimated in the bed-seated position at the end of the visit compared with the recommended chair-seated position in treated and untreated patients with EH, in particular in elderly obese women with mild hypertension. PMID- 15752954 TI - Assessment of central and peripheral arterial stiffness: studies indicating the need to use a combination of techniques. AB - BACKGROUND: Several new techniques exist for measuring arterial stiffness, but their association with central pulse wave velocity (PWV), an established measure of central arterial stiffness, has not been compared in the same study. The aim of this study was to compare the agreement of the new techniques with central PWV. METHODS: Fifteen men with coronary artery disease (59 +/- 2 years of age) and 15 healthy men (28 +/- 1 years of age) were recruited. The following measures were performed in a random sequence and repeated after 1 week: central and distal pulse wave velocity (PWV), large and small artery compliance (C1 and C2, respectively), and stroke volume/pulse pressure (SV/PP) (measured by HDI/PulseWave CR-2000), augmentation index (AIx) and central pulse pressure (CPR) (Sphygmocor), stiffness index (SI) (Micro Medical Pulse Trace), systemic arterial compliance (SAC) (area method), and brachial PP (Dinamap). Methods were compared using correlation coefficients and Bland-Altman analysis. RESULTS: Although all measures of arterial stiffness except PWV correlated significantly with central PWV (P < .01 for each), Bland-Altman analysis showed poor agreement (confidence interval [CI] >3 Z-scores) between central PWV and C1, C2, SV/PP, and SAC). There was good agreement (CI > 2 Z-scores) between central PWV and SI, AIx, CPP, and brachial PP. The coefficient of variation was lowest with central PWV (7.6%), brachial PP (8.0%), and SV/PP (8.6%) and was significantly higher (P < .05) in increasing order with C1 (11.3%), C2 (15.6%), SI (17.8%), SAC (19.3%), AIx (22.4%), and CPP (25.3%). CONCLUSIONS: Based on our study findings, C1, C2, SV/PP, and SAC show poor agreement with central PWV, an established measure of central arterial stiffness. Indices of this type should therefore be useful in providing a more complete understanding of arterial stiffness. In comparison, SI, AIx, and CPP are more closely related to central arterial stiffness. PMID- 15752955 TI - Low mineral intake is associated with high systolic blood pressure in the Third and Fourth National Health and Nutrition Examination Surveys: could we all be right? AB - Analysis of the first National Health and Nutrition Examination Survey (NHANES) in 1984 revealed that a dietary pattern low in mineral intake, specifically calcium, potassium, and magnesium, was associated with hypertension in American adults. Using more recent survey data from NHANES III and NHANES IV, we re examined the validity of this relationship. Blood pressure (BP) and nutrient intake data from 10,033 adult participants in NHANES III and 2311 adults in NHANES IV revealed findings similar to those of the earlier analysis, demonstrating that the association between inadequate mineral consumption and higher BP is valid and has persisted over two decades. Exploring this relationship further, we separated untreated hypertensive persons by hypertension type (systolic, diastolic, or both), and observed that the BP effect of low mineral intake was most pronounced in those with only systolic hypertension. We also observed that sodium intake was significantly lower in the systolic hypertension group and significantly higher in the diastolic hypertension group compared with the other groups. The nutrient pattern in the combined hypertension group was similar to that of the normotensive group. These findings may help to explain the inconsistent responses generally observed in dietary intervention studies, and they highlight the possible importance of tailored nutritional recommendations for hypertension based on hypertension category and individual dietary practices. Although randomized controlled trials are needed to characterize further the relationship between nutrient intake and hypertension type, these findings indicate that dietary management of hypertension may be more effective if the focus is on the overall nutritional profile rather than single nutrient intake as currently recommended for most patients. PMID- 15752956 TI - Impact of a social marketing media campaign on public awareness of hypertension. AB - BACKGROUND: Barriers to high blood pressure (BP) awareness and control are exacerbated by poor knowledge of the consequences and uncertainty regarding how to and who should direct care. We developed a social marketing hypertension awareness program to determine baseline awareness, knowledge, and treatment behavior, and then studied the impact of a targeted, media intervention among randomly surveyed adults at risk in a representative urban community compared to a control community immediately and 6 months after the intervention. METHODS: The program consisted of three random-digit telephone surveys conducted in two mid sized Ontario cities to determine high BP awareness, knowledge, and treatment behavior. Using baseline knowledge and attitudes toward high BP in both communities, a social marketing awareness strategy and mass media intervention campaign incorporating television, radio, print, direct to patient, and interactive techniques was developed and implemented in the test city only. Both test and control cities were resurveyed immediately after and at 6 months post media intervention to detect change and decay. RESULTS: A sample of 6873 men and women more than 35 years old who were aware of their high BP demonstrated a high prevalence of high BP in the general population ( approximately 34% in both communities). At baseline this population had poor knowledge of their own BP numbers and poor understanding of the diseases related to high BP. Although few considered high BP a health concern, they had good understanding of lifestyle interventions for high BP prevention and control. The number of the respondents who claimed to have high BP increased immediately after intervention in the test city (38%; P < .02), whereas the number of respondents who were treated and uncontrolled decreased (P < .05) compared to control. There was a significant increase in patients' knowledge of consequences and in their perception that they were most responsible for high BP control in the test city (P < .005) compared to control. At 6 months, no further changes were observed in those claiming to have high BP in either city, whereas decay to baseline in those treated but not controlled and those claiming responsibility for their BP control was observed in the test city. No changes were observed in the control city accept for an increase from baseline to 6 months in the percentage claiming to be treated but uncontrolled. We were unable to determine whether the increase in number treated but uncontrolled was due to a higher treatment rate, similar treatment rate but more patients being uncontrolled, or a combination of these scenarios. CONCLUSIONS: High BP is very prevalent in adults and knowledge of lifestyle options for management is encouraging. In the short-term, although our media awareness program increased the number of respondents claiming to have high BP and patient self-efficacy for BP control, this was not maintained. We did not change knowledge of consequences or importantly, the health importance of BP control among those at risk. Hence, in addition to a mass media campaign, attention should be focused on dissemination of awareness knowledge information through medical professionals at the point of care. PMID- 15752957 TI - Systematic review of the effect of daily alcohol intake on blood pressure. AB - Numerous epidemiologic investigations have found an association between moderate intake of alcohol and increased blood pressure (BP). However, in controlled clinical studies that directly tested the effects of alcohol intake on BP, findings are inconsistent, perhaps because of differences in duration of alcohol use and the timing of BP measurements. In this setting, we performed a systematic review of trials that measured BP after a period of sustained alcohol intake (defined as daily intake of at least one alcoholic drink daily) in one group and that also had a control group of individuals who consumed no alcohol. Nine studies met the entrance criteria. The review demonstrated a significant rise in systolic blood pressure (SBP) and diastolic BP (DBP) of 2.7 mm and 1.4 mm Hg, respectively, after alcohol intake. An early effect of alcohol leading to a reduction BP (in the hours after exposure) and a later effect (next day) of raising BP led to smaller differences in the net effect of alcohol on BP when ambulatory BP monitoring measurements were compared with casual office- or clinic based measurements. Our findings may have important implications for interpreting studies measuring the effect of alcohol on BP as well as for regular clinical care. These findings indicate that the timing of BP measurements after alcohol intake has a substantial effect on the magnitude and perhaps even the direction of BP change. PMID- 15752958 TI - Role of angiotensin receptor blockers as monotherapy in reaching blood pressure goals. AB - The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure emphasizes the urgent need to lower blood pressure (BP) to a goal of <140/90 mm Hg in patients with uncomplicated hypertension and to <130/80 mm Hg in high-risk patients, such as those with diabetes mellitus or chronic kidney disease, to prevent cardiovascular disease morbidity and mortality. Consequently, a meaningful measure of the efficacy of an antihypertensive therapy is its ability to achieve BP reduction to below the recommended BP goals. Angiotensin II receptor blockers (ARB) are highly effective antihypertensive agents with excellent tolerability profiles similar to those of placebo. A literature search using MEDLINE, EMBASE, and BIOSIS to identify studies reporting data on the percentage of patients attaining BP goals found that monotherapy with an ARB can generally result in the attainment of the diastolic BP (DBP) goal of <90 mm Hg in approximately 50% of hypertensive patients. However, to our knowledge, the attainment of the systolic BP (SBP) and combined SBP/DBP goals with ARB monotherapy has not been reported. Therefore, a secondary analysis of BP efficacy data from a published study that directly compared recommended starting doses of four currently marketed ARB was performed to assess combined SBP and DBP goal attainment. This analysis showed that the percentage of patients achieving the combined SBP/DBP goal rate of <140/90 mm Hg was highest with olmesartan medoxomil (32.4%) compared with recommended starting doses of losartan potassium (16.1%), valsartan (14.5%), or irbesartan (25.9%). Optimal ARB monotherapy can achieve recommended BP goals in a significant proportion of hypertensive patients. However, the majority of hypertensive patients will require combination therapy with two or more antihypertensive agents. PMID- 15752959 TI - Increased calcium/magnesium ratio in young and old spontaneously hypertensive rats: Pathogenetic factor in the development of hypertension and vascular fibrosis. PMID- 15752961 TI - Correlation of neurological manifestations of Takayasu's arteritis with cerebral angiographic findings. AB - For categorizing the presenting neurological symptoms in Takayasu's arteritis (TA) and correlating to the imaging findings of cerebral angiography, we retrospectively reviewed the angiographies of 27 patients presented with stroke, transient ischemic attack (TIA), visual disturbance, and/or dizziness. Neurological manifestations of TA resulted from decreased blood flow due to a steno-occlusive lesion and/or shifting of blood flow, thromboembolism, or hypertension. The pattern analysis of neurological manifestations related to specific arterial involvement is mandatory in understanding neurological manifestations of this systemic disease. PMID- 15752962 TI - Diffusion-weighted imaging with calculated apparent diffusion coefficient in intracranial hemorrhagic lesions. AB - In the literature published so far, measurement of values of the apparent diffusion coefficient (ADC) using an echo-planar imaging (EPI) technique in intracranial hemorrhagic lesions show no uniform results. Furthermore, no data exist for bleedings into intracranial lesions. We investigated the ADCs of 18 intracranial hemorrhagic lesions of different stages using echo-planar diffusion weighted imaging (DWI). The ADC values measured in the hemorrhagic lesions ranged from 1.42 x 10(-3) to 0.22 x 10(-3) mm(2)/s. There were no significant differences between the ADC values in the hemorrhagic lesions and the contralateral white matter (P=.39). A differentiation between the lesions only with the ADC value was not possible as well. Using EPI DWI in intracranial hemorrhagic lesions of different stages, no reliable ADC values were found and a dependable differentiation between the lesions is not possible. PMID- 15752963 TI - Patterns of brain activation in patients with mild Alzheimer's disease during performance of subtraction: an fMRI study. AB - We performed a functional MRI (fMRI) study to compare the difference of activation between healthy aged people and patients with mild Alzheimer's disease (AD) during performance of subtraction. Nine patients with mild AD and nine healthy aged volunteers were recruited in this study. The analysis of fMRI data revealed that brain activation is decreased in several regions in AD patients in comparison with healthy participants. But in the right inferior prefrontal lobe, activation is greater in patients than in the controls. We believe that our findings will help the understanding mechanism of neuronal activity in AD. PMID- 15752964 TI - Percutaneous nucleotomy: preliminary communication on a decompression probe (Dekompressor) in percutaneous discectomy. Ten case reports. AB - For the past years, new therapeutic options have been proposed secondary to the progress in the interventional imaging. The purpose of this article is to report a preliminary study on the use of the 1.5 mm "Dekompressor probe" (Stryker, Kalamazoo, MI, USA) in the percutaneous discectomy by decompression. We have conducted, from September 2003 to March 2004, a retrospective review on 10 patients (mean age of 49.8), chosen at random, presenting a nonextruded herniated disc resisting all medical treatments. The procedure is CT guided or performed under fluorosocopy, a discography is performed by introducing the probe at the level of the protruded disc. After local anesthesia, an incision of a few millimeters is performed, allowing the introduction of a coxial trocar to the level of the disc. The decompression probe is then introduced. No technical failure occurred. The results in our series show, in eight patients, a satisfactory result with a decrease of the initial VAS of more than 70% and a complete elimination of the medical therapy. Our preliminary results are encouraging but should also be confirmed by a multicentric based on a large series, and the criteria of inclusion or exclusion must be strictly respected to obtain satisfactory clinical results. PMID- 15752965 TI - Assessment of renal cell carcinoma after cryoablation using contrast-enhanced gray-scale ultrasound: a case series. AB - We performed contrast-enhanced gray-scale harmonic ultrasound (CUS) in three patients with renal cell carcinoma after cryoablation. CUS showed the decreased enhancement similar to the findings of CT or MRI. It identified feeding vessels more distinctly and found a larger number compared with CT or MRI. A persistent feeder in one case was related to recurrence. CUS may potentially be an alternative to CT and MRI and a useful method in the prediction of recurrent and residual tumors. PMID- 15752966 TI - Posttransplant symptomatic lymphocele treatment with percutaneous drainage and ethanol sclerosis: long-term follow-up. AB - PURPOSE: Lymphocele following kidney transplantation is a well-known complication. We present here the long-term results of percutaneous drainage and ethanol sclerosis in posttransplant lymphoceles. PATIENTS AND METHODS: Between October 1988 and September 2003, 18 patients with symptomatic lymphoceles were treated with percutaneous transcatheter ethanol sclerosis among 300 renal transplants followed up. Catheters were positioned under ultrasonography (US) guidance. Cavity dimensions and extravasation were evaluated by pouch graph. The day after catheter insertion, sclerotherapy with ethanol was applied. Daily output and probable complications were followed up on periodic controls, and the alcohol was instilled when required. RESULT: The mean initial size of the lymphoceles was 6 x 8 x 10 cm (range 3 x 4 x 6 to 7 x 12 x 21 cm), with a mean initial volume of 230 ml (range 40-1340 ml). Incomplete resolution was achieved in all patients after sclerotherapy in the first week (100%). The mean duration of therapy was 17 days (range 4-45 days). The mean therapy sitting was 12 (4-21), and the average alcohol volume in one sitting was 30 ml. One recurrence and one graft lost were seen in this series. Minor complications like discomfort and mild temperature elevation were determined in 10 patients. In one case, temporary double-J stent was required to alleviate the increased pressure in collecting system of transplanted kidney. No late complication was occurred. CONCLUSION: As a less invasive method, percutaneous transcatheter ethanol sclerosis and drainage in the treatment of symptomatic lymphocele is a simple, safe and cost-effective method. Other treatment choices, like open or laparoscopic fenestration, can be used in case of failure. PMID- 15752967 TI - Magnetic resonance delineation of the anterior cruciate ligament of the knee: flexed knee position within a surface coil. AB - The purpose of this study was to evaluate the effect of the knee position at three different flexion angles in magnetic resonance (MR) delineation of the anterior cruciate ligament (ACL) in the knee and to determine the optimal knee position. Thirteen knees of normal volunteers were examined at 15 degrees, 30 degrees, and 45 degrees of flexion with a surface coil, and three sets of obtained oblique sagittal MR images were evaluated by four observers. MR images at 30 degrees of knee flexion most clearly delineate compared with those at 15 degrees and 45 degrees of knee flexion. We recommended examining the knee in 30 degrees of flexion. PMID- 15752968 TI - Efficacy of CT-guided percutaneous needle biopsy in the diagnosis of malignant lymphoma at first presentation. AB - PURPOSE: The aim of this study was to evaluate retrospectively the accuracy and reliability of CT-guided percutaneous biopsy as an alternative to surgical biopsy in a selected population of patients without superficial enlarged lymph nodes and a final diagnosis of malignant lymphoma at first presentation. METHODS: The results of 145 CT-guided needle biopsies in 137 patients with malignant lymphoma at its first presentation and without superficial enlarged lymph nodes were analyzed retrospectively. Biopsies were performed in 24 patients with Hodgkin's disease (HD) and 113 with non-Hodgkin lymphoma (NHL). Factors such as patient's sex, age, type of lymphoma and biopsy site were evaluated to detect factors that could influence the success rate of the procedure. RESULTS: Biopsy specimens were diagnostic in 101 of the 113 patients with NHL and in 18 of the 24 patients with HD. Repeating of a previously nondiagnostic biopsy was successful in 7 out of 13 patients with NHL. No positive results were obtained, repeating the inconclusive biopsy in six patients with HD. CONCLUSIONS: Our results suggest that percutaneous CT-guided biopsy is a useful and reliable tool in the diagnosis and classification of malignant lymphomas in patients without superficial lymphadenopathy and can be considered as an alternative to surgical sampling. However, little advantages were obtained, repeating previously inconclusive biopsies: In these cases, surgical sampling is mandatory. PMID- 15752969 TI - Nasopharyngeal carcinoma: the impact of CT-scan and of MRI on staging, radiotherapy treatment planning, and outcome of the disease. AB - The present study is a critical review of the role of computed tomography (CT) and magnetic resonance imaging (MRI) in the diagnosis and therapy of nasopharyngeal cancer. It is estimated that following CT-scan/MRI, T,N staging of the disease changes in about half of cases, which results in important adjustments of the radiotherapy treatment planning, both in terms of radiation field dimensions and of dose specifications. The development of novel technology with CT-scan-simulated 3-D conformal or intensity-modulated radiotherapy (IMRT) planning is gradually becoming the standard therapy for nasopharyngeal carcinoma (NPC). CT-scan/MRI is also of value in detecting massive parapharyngeal involvement, low-neck lymphadenopathy or fixation of nodes onto adjacent structures, which are important features indicating the necessity to integrate chemotherapy or surgical neck dissection in the overall treatment policy. CT positron emission tomography (PET) scan is recently under evaluation for the identification of the most active tumor regions, which will allow a biological radiotherapy planning (RTP) using IMRT techniques. PMID- 15752970 TI - Malignant fibrous histiocytoma of the female breast: a case report. AB - Malignant fibrous histiocytoma (MFH) of the breast is very rare. Sonographic findings of MFH of the breast have not been reported before. We herein report a case of a 46-year-old woman with a histologically proved MFH of her right breast. Ultrasound examination revealed a 6-cm tumor mass with solid and cystic components, which were attributed after histological examination to hypercellularity and hemorrhage within the tumor. Histopathological diagnosis was compatible with MFH of the breast. PMID- 15752971 TI - Descending necrotizing mediastinitis: a rare pathology to keep in mind. AB - The spreading of a cervical infection to the mediastinum is a complication rarely observed, and its prognosis is still very severe. The infectious spreading follows known anatomical tracts, leading to the invasion of definite mediastinal spaces. The cervicothoracic scanning is the ideal procedure leading to the diagnosis, the localization of the infectious sites, and to the demonstration of an abscess and the presence of air in the mediastinum. We report a case of a patient in whom the early use of specific imaging and of proper aggressive surgical intervention did not allow the elimination of a fatal outcome. PMID- 15752972 TI - CT characteristics of cervical Castleman's disease. AB - Castleman's disease is an uncommon disorder of benign lymph-node hyperplasia. Among all the lesions, less than 10% occurs in the neck. Our case differs from those cases previously reported. Multiple lesions were found on both sides of the neck. Contrast-enhanced CT scan showed dotty and stripy enhancement around these lesions, and rim-like enhancement was also found on delayed scan at 5 min as well. The possibility of Castleman's disease should be considered in multiple cervical masses that show well-defined marked and homogeneous enhancement, especially when stripy enhancement is found around the lesions simultaneously. PMID- 15752973 TI - Perforated colonic diverticulitis mimicking urachal carcinoma: computed tomography and magnetic resonance findings. AB - We report on a case of perforated diverticulitis of the sigmoid colon, which mimicked urachal carcinoma, and discuss its radiological findings. PMID- 15752974 TI - Leucine-rich nuclear-export signals: born to be weak. AB - CRM1 mediates the nuclear export of proteins exposing leucine-rich nuclear-export signals (NESs). Most NESs bind to CRM1 with relatively low affinity. Recently, higher-affinity NESs were selected from a 15-mer random peptide library. Unexpectedly, complexes between high-affinity NESs and CRM1 accumulate at the cytoplasmic filaments of the nuclear pore complex (NPC). This finding suggests that high-affinity NES binding to CRM1 impairs the efficient release of export complexes from the NPC, explaining why leucine-rich NESs have evolved to be weak. PMID- 15752975 TI - Vacuolar proteases livening up programmed cell death. AB - The molecular identity of the key executioners involved in controlling plant programmed cell death (PCD) has been elusive. In a recent paper published in Science, Hatsugai and coworkers reported that a well-characterized protease called VPE from the plant cell vacuole can cleave caspase-specific substrates and is required for cell death activation by tobacco mosaic virus. This work provides clear evidence for the importance of the vacuole in plant PCD and a novel regulatory function for this organelle as well as for VPE proteases. PMID- 15752976 TI - More than just proliferation: Myc function in stem cells. AB - Adult stem cells are essential to maintain regenerative tissues such as skin epidermis, gastrointestinal mucosa or the hematopoietic system. Recent studies in mice suggest that the transcription factor and oncoprotein c-Myc has unexpected functions during both self-renewal and the differentiation of stem and early progenitor cells, particularly in interactions between stem cells and the local microenvironment or "niche". By incorporating recent findings on Myc and hematopoietic stem cells we propose a model in which "resting" hematopoietic stem cells are "activated" to self-renew and to differentiate at the interface between the niche and non-niche microenvironments. PMID- 15752977 TI - Tumor cells caught in the act of invading: their strategy for enhanced cell motility. AB - Invasion of neighboring extracellular matrix tissue, the lymphatic system and blood vessels is a key element of tumor cell metastasis in many epithelial tumors. Understanding the cell motility pathways that contribute to invasion can provide new approaches and targets for anticancer therapy. The recent convergence of technologies for expression profiling and intravital imaging has revealed the identities of some of the genes that contribute to motility and chemotaxis of cancer cells in tumors. In particular, the genes encoding a minimum motility machine are coordinately upregulated in tumor cells collected by an in vivo invasion assay. These results support a "tumor microenvironment invasion model" and provide new target opportunities for cancer therapy. PMID- 15752978 TI - Turning cells red: signal transduction mediated by erythropoietin. AB - Erythropoietin (EPO) is the crucial cytokine regulator of red blood-cell production. Since the discovery of EPO in 1985 and the isolation of its cognate receptor four years later, there has been significant interest in understanding the unique ability of this ligand-receptor pair to promote erythroid mitogenesis, survival and differentiation. The development of knockout mice has elucidated the precise role of the ligand, receptor and downstream players in murine erythroid development. In this review, we summarize EPO-mediated signaling pathways and examine their significance in vivo. PMID- 15752979 TI - Site selection for the cleavage furrow at cytokinesis. AB - The question of how the site for division of the cytoplasm is determined at the end of mitosis has been studied for over a century, and it remains an active, controversial and fascinating problem in cell biology. This problem draws on the use of several model cell types, with the goal of understanding and identifying how the cell cycle regulates signals between the mitotic apparatus and the cell cortex. Studies in different cell types and using a vast array of techniques reveal different answers: these might reflect differences in experimental approaches, multiple and redundant mechanisms and, importantly, diversity in biology. In this article (which is part of the Cytokinesis series), we present a summary and critique of the major models for the roles of the mitotic apparatus microtubules in stimulating furrow formation at cytokinesis. PMID- 15752980 TI - Regulation of innate immunity by Rho GTPases. AB - Leukocytes are key cellular components of innate immunity. These phagocytic cells respond to bacteria at sites of infection through chemotactic sensing and directed motility regulated by Rho GTPases. The development of sensitive probes of Rho GTPase dynamics has provided insights into the temporal and spatial aspects of GTPase regulation during chemotaxis and subsequent microbial phagocytosis. The resulting destruction of ingested bacteria by means of reactive oxygen species (ROS) depends on a Rac-regulated "molecular switch" that is modulated by antagonistic crosstalk involving Cdc42. Recent studies of leukocytes derived from Rac1- and Rac2-knockout mice have shown that these highly homologous GTPases have unique biological roles. An understanding of the biochemical basis for such distinct activities should provide novel insights into the molecular details of Rho GTPase function and regulation in innate immunity. PMID- 15752981 TI - Delta-catenin at the synaptic-adherens junction. AB - Delta-catenin belongs to the p120-catenin (p120(ctn)) protein family, which is characterized by ten, characteristically spaced Armadillo repeats that bind to the juxtamembrane segment of the classical cadherins. Delta-catenin is the only member of this family that is expressed specifically in neurons, where it binds to PDZ domain proteins in the post-synaptic compartment. As a component of both adherens and synaptic junctions, delta-catenin can link the adherens junction to the synapse and, thereby, coordinate synaptic input with changes in the adherens junction. By virtue of its restriction to the post-synaptic area, delta-catenin creates an asymmetric adherens junction in the region of the synapse. The crucial nature of the specialized function of delta-catenin in neurons is demonstrated by a targeted gene mutation, which causes deficits in learning and in synaptic plasticity. Taken together, recent evidence indicates that delta-catenin is a sensor of synaptic activity and implements activity-related morphological changes at the synapse. PMID- 15752982 TI - RS domains contact the pre-mRNA throughout spliceosome assembly. AB - SR proteins are essential metazoan splicing factors that contain an RNA-binding domain and an arginine/serine-rich domain that functions to promote assembly of the spliceosome. The prevailing model over the past several years suggests that the RS domains function as protein-interaction domains. However, two new papers from Green et al. demonstrate that these RS domains directly contact the pre-mRNA within the functional spliceosome. The sequential character of these contacts suggests that RS domain interactions with RNA promote spliceosome assembly. PMID- 15752983 TI - From genes to machines: DNA nanomechanical devices. AB - The structural properties that enable DNA to serve so effectively as genetic material can also be used for other purposes. The complementarity that leads to the pairing of the strands of the DNA double helix can be exploited to assemble more complex motifs, based on branched structures. These structures have been used as the basis of larger 2D and 3D constructions. In addition, they have been used to make nanomechanical devices. These devices range from DNA-based shape shifting structures to gears and walkers, a DNA-stress gauge and even a translation device. The devices are activated by mechanisms as diverse as small molecules, proteins and, most intriguingly, other molecules of DNA. PMID- 15752984 TI - Expanding functional diversity of the coactivators. AB - Nuclear receptors (NR) function as ligand-regulated transcription factors that transduce hormonal signals from steroid hormones and other lipophillic ligands. NR-mediated transcription depends on coactivators, a diverse group of proteins that affect the transcriptional machinery in a variety of ways such as via their associated enzymatic activities as histone acetyltransferases, methyltransferases, ubiquitin ligases or as agents that integrate signaling via kinase-signaling pathways. Coactivators have various roles in the transcriptional process (i) as molecules that influence key points in the different stages of transcription, (ii) as integrators of environmental growth-factor and steroid hormone signals, and (iii) as agents of carcinogenesis. PMID- 15752985 TI - Iron-sulfur-protein biogenesis in eukaryotes. AB - Iron-sulfur (Fe-S) clusters (ISCs) are versatile, ancient co-factors of proteins that are involved in electron transport, enzyme catalysis and regulation of gene expression. The synthesis of ISCs and their insertion into apoproteins involves the function of complex cellular machineries. In eukaryotes, the mitochondrial ISC-assembly machinery is involved in the maturation of all cellular iron-sulfur proteins. A mitochondrial export machinery and a recently discovered cytosolic assembly system specifically participate in the maturation of cytosolic and nuclear iron-sulfur proteins. Of the approximately 20 assembly components, more than ten are encoded by essential genes, which indicates that the process is indispensable for life. Mutations in two of the assembly components lead to neurological diseases. The essential character of Fe-S-protein biogenesis in eukaryotes and its importance for human disease identifies this evolutionary ancient process as one of the most important biosynthetic pathways of life. PMID- 15752986 TI - Multifaceted roles of glycolytic enzymes. AB - Although glycolysis is a biochemical pathway that evolved under ancient anaerobic terrestrial conditions, recent studies have provided evidence that some glycolytic enzymes are more complicated, multifaceted proteins rather than simple components of the glycolytic pathway. These glycolytic enzymes have acquired additional non-glycolytic functions in transcriptional regulation [hexokinase (HK)-2, lactate dehydrogenase A, glyceraldehyde-3-phosphate dehydrogenase (GAPD) and enolase 1], stimulation of cell motility (glucose-6-phosphate isomerase) and the regulation of apoptosis (glucokinase, HK and GAPD). The existence of multifaceted roles of glycolytic proteins suggests that links between metabolic sensors and transcription are established directly through enzymes that participate in metabolism. These roles further underscore the need to consider the non-enzymatic functions of enzymes in proteomic studies of cells and tissues. PMID- 15752987 TI - The RIP kinases: crucial integrators of cellular stress. AB - Since the discovery of the first member ten years ago, the receptor-interacting protein (RIP) family kinases have emerged as essential sensors of cellular stress. The different members integrate both extracellular stress signals transmitted by various cell-surface receptors and signals emanating from intracellular stress. The cascades of events initiated by activated RIPs are complex. Not only are pro-survival, inflammatory and immune responses triggered by RIP kinases via the activation of transcription factors such as NF-kappaB and AP-1, but opposing, death-inducing programs can also be initiated by the RIP kinases. Hence, RIP kinases are crucial regulators of cell survival and cell death. PMID- 15752989 TI - The consequences of gene and genome duplication in plants. PMID- 15752988 TI - Historical article: Titration studies and the structure of DNA. AB - During the period of 1946-1956 ideas on the structure of the nucleic acids, especially DNA, were profoundly influenced by physico-chemical studies and, in particular, by electrometric titration studies pioneered by J. Masson Gulland and D.O. Jordan. It was eventually conclusively inferred when such studies had been sufficiently refined that: in DNA, the only hydrogen bonding that occurred was between the 1:6 amino groups of adenine and cytosine, and the -NH-CO "enolic" groups of thymine and guanine; the polynucleotide chains of DNA were not branched (or interrupted); and the internally hydrogen-bonded double-helical structure was stable both in aqueous solution and in the fibrous state. PMID- 15752990 TI - The evolutionary dynamics of plant duplicate genes. AB - Given the prevalence of duplicate genes and genomes in plant species, the study of their evolutionary dynamics has been a focus of study in plant evolutionary genetics over the past two decades. The past few years have been a particularly exciting time because recent theoretical and experimental investigations have led to a rethinking of the classic paradigm of duplicate gene evolution. By combining recent advances in genomic analysis with a new conceptual framework, researchers are determining the contributions of single-gene and whole-genome duplications to the diversification of plant species. This research provides insights into the roles that gene and genome duplications play in plant evolution. PMID- 15752991 TI - Evolving disease resistance genes. AB - Defenses against most specialized plant pathogens are often initiated by a plant disease resistance gene. Plant genomes encode several classes of genes that can function as resistance genes. Many of the mechanisms that drive the molecular evolution of these genes are now becoming clear. The processes that contribute to the diversity of R genes include tandem and segmental gene duplications, recombination, unequal crossing-over, point mutations, and diversifying selection. Diversity within populations is maintained by balancing selection. Analyses of whole-genome sequences have and will continue to provide new insight into the dynamics of resistance gene evolution. PMID- 15752992 TI - Polyploidy and genome evolution in plants. AB - Genome doubling (polyploidy) has been and continues to be a pervasive force in plant evolution. Modern plant genomes harbor evidence of multiple rounds of past polyploidization events, often followed by massive silencing and elimination of duplicated genes. Recent studies have refined our inferences of the number and timing of polyploidy events and the impact of these events on genome structure. Many polyploids experience extensive and rapid genomic alterations, some arising with the onset of polyploidy. Survivorship of duplicated genes are differential across gene classes, with some duplicate genes more prone to retention than others. Recent theory is now supported by evidence showing that genes that are retained in duplicate typically diversify in function or undergo subfunctionalization. Polyploidy has extensive effects on gene expression, with gene silencing accompanying polyploid formation and continuing over evolutionary time. PMID- 15752993 TI - Arabidopsis epigenetics: when RNA meets chromatin. AB - Recent work in plants and other eukaryotes has uncovered a major role for RNA interference in silent chromatin formation. The heritability of the silent state through multiple cell division cycles and, in some instances, through meiosis is assured by epigenetic marks. In plants, transposable elements and transgenes provide striking examples of the stable inheritance of repressed states, and are characterized by dense DNA methylation and heterochromatin histone modifications. Arabidopsis is a useful higher eukaryotes model with which to explore the crossroads between silent chromatin and RNA interference both during development and in the genome-wide control of repeat elements. PMID- 15752994 TI - Advances in plant chromosome identification and cytogenetic techniques. AB - Recent developments that improve our ability to distinguish slightly diverged genomes from each other, as well as to distinguish each of the nonhomologous chromosomes within a genome, add a new dimension to the study of plant genomics. Differences in repetitive sequences among different species have been used to develop multicolor fluorescent in situ hybridization techniques that can define the components of allopolyploids in detail and reveal introgression between species. Bacterial artificial chromosome probes and repetitive sequence arrays have been used to distinguish each of the nonhomologous somatic chromosomes within a species. Such karyotype analysis opens new avenues for the study of chromosomal variation and behavior, as well as for the localization of individual genes and transgenes to genomic position. PMID- 15752995 TI - Updating the 'crop circle'. AB - Comparative analyses unravel the relationships between genomes of related species. The most comprehensive comparative dataset obtained to date is from the grass family, which contains all of the major cereals. Early studies aimed to identify chromosomal regions that have remained conserved over long evolutionary time periods, but in recent years, researchers have focused more on the extent of colinearity at the DNA-sequence level. The latter studies have uncovered many small rearrangements that disturb colinearity in orthologous chromosome regions. In part, genomes derive their plasticity from genome- and gene-amplification processes. Duplicated gene copies are more likely to escape selective constraints and thus move to other regions of the genome, where they might acquire new functions or become deleted. These rearrangements will affect map applications. The most popular applications, especially since the complete rice genomic sequence has been available, are the use of comparative data in the generation of new markers to tag traits in other species and to identify candidate genes for these traits. The isolation of genes underlying orthologous traits is the first step in conducting comparative functional studies. PMID- 15752996 TI - Plant biotechnology. PMID- 15752997 TI - Autophagic recycling: lessons from yeast help define the process in plants. AB - The autophagic engulfment of cytoplasm and organelles and their delivery to the vacuole have long been speculated to play an essential role in bulk protein turnover in plants. Until recently, however, the importance and the mechanism(s) of action of these processes have remained obscure. Aided by the discovery of numerous orthologs of the yeast AUTOPHAGY (ATG) protein system in Arabidopsis, significant advances have been now made in understanding these processes. Both reverse genetic analyses of the Arabidopsis ATG genes and the use of the encoded proteins as cytological markers have confirmed the presence of autophagy in plants and have demonstrated its importance in nutrient recycling, especially during senescence and growth under carbon- or nitrogen-limiting conditions. PMID- 15752998 TI - Flux an important, but neglected, component of functional genomics. AB - Genomics approaches aimed at understanding metabolism currently tend to involve mainly expression profiling, although proteomics and steady-state metabolite profiling are increasingly being carried out as alternative strategies. These approaches provide rich information on the inventory of the cell. It is, however, of growing importance that such approaches are augmented by sophisticated integrative analyses and a higher-level understanding of cellular dynamics to provide insights into mechanisms that underlie biological processes. We argue the need for, and discuss theoretical and practical aspects of, the determination of metabolic flux as a component of functional genomics. PMID- 15752999 TI - Dormancy release, ABA and pre-harvest sprouting. AB - Seed dormancy is an adaptive trait that enables the seeds of many species to remain quiescent until conditions become favorable for germination. Dormancy is normally initiated during seed maturation and maintained to seed maturity. In mature seeds, the loss of dormancy may be gradual (after-ripening) or can be terminated by chilling and other environmental triggers. Dormancy is an important trait for many important crop species: it inhibits pre-harvest spouting or vivipary, a widespread problem in many regions of the world. Too much dormancy, however, can lead to non-uniform germination in the field. Recent progress has been made in understanding the role of abscisic acid metabolism and dormancy release in both model plants and crop species. Advances in our understanding of the molecular mechanisms that are involved in dormancy, along with approaches using quantitative genetics, will provide new strategies through which the desired level of dormancy can be introduced into crop species. PMID- 15753001 TI - Metabolic engineering of new fatty acids in plants. AB - Metabolic engineering of plants to express high levels of new fatty acids that are of nutritional and industrial importance has proven to be highly challenging. Significant advances have been made recently, however, particularly in the development of the first plant oils to contain long-chain polyunsaturated fatty acids, such as arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. Methods of increasing the accumulation of Delta12-modified fatty acids synthesized by transgenically expressed FAD2-like enzymes have also been investigated. Biochemical analyses of plants that express these introduced fatty acid metabolic pathways have highlighted the central importance of ensuring the removal of novel fatty acids from their site of synthesis on phosphatidylcholine to enable their further modification, exclusion from membrane lipids and accumulation in seed triacylglycerols. PMID- 15753000 TI - Plant-based material, protein and biodegradable plastic. AB - Fibrous proteins from spiders, proteins with synthetic multiple repeats and mammalian structural proteins such as collagen have been produced in transgenic plants. Recent advances in the production of biodegradable plastic in plants also show the potential of molecular farming for research into and production of materials. Selection of a growing variety of such products, optimization of expression, and the development of effective purification strategies will further promote this growing field of biotechnology. PMID- 15753002 TI - Towards the rational design of cereal starches. AB - A major challenge in cereal biotechnology is to achieve the rational design of renewable polymers to meet specific requirements for improving human health, nutrition, and food quality, to increase the energy supply, and to provide safer and more profitable industrial inputs. The field of starch synthesis research has advanced at a rapid pace over the past decade, and many core observations about the pathway are well established over a range of species. Owing to the complexity of the starch-synthesis process, in which suites of enzymes act at the interface between soluble and insoluble phases, the rational design of starch granules with specific functionality is still in its infancy. Our fundamental biochemical knowledge of starch biosynthesis has recently advanced, and this new information could be exploited to create novel variability in carbohydrate polymers in cereal grains. We propose two strategies for moving more rapidly towards truly rational design of starch. First, the focusing of fundamental research on processes that are involved in the regulation of starch synthesis and granule assembly. Second, the development of iterative strategies, exploiting new molecular genetics tools, to screen for desired properties in high-throughput systems. PMID- 15753003 TI - TILLING without a plough: a new method with applications for reverse genetics. AB - TILLING (Targeting Induced Local Lesions IN Genomes) is a powerful reverse genetic technique that employs a mismatch-specific endonuclease to detect induced or natural DNA polymorphisms. Its advantages over other reverse genetic techniques include its applicability to virtually any organism, its facility for high-throughput and its independence of genome size, reproductive system or generation time. TILLING is currently being used for the detection of both induced and natural variation in several plant species. PMID- 15753004 TI - RNA silencing and antiviral defense in plants. AB - Much progress has been made recently in identifying the molecular components of RNA silencing in plants, and in understanding their roles in the biogenesis of small interfering RNAs and microRNAs, in RNA-directed DNA methylation, and in RNA mediated antiviral defense. However, many crucial questions remain unanswered. What are the molecular bases of sense and antisense transgene-mediated silencing? Why does silencing only appear to spread through transgenes? Plant viruses encode silencing suppressors to counteract host RNA silencing, and some of these suppressors affect microRNA accumulation and function and hence normal plant development. Is viral pathogenicity determined, partly or entirely, by their silencing suppressor activity? PMID- 15753005 TI - Bioprospecting in plants for engineered proteins. AB - For more than two decades, bioengineered plants have produced protein therapeutics for human and animal use. Almost all proteins produced by other existing systems, including antibodies, vaccines and plasma proteins, have now been manufactured in plants. Considering the limitations of microbial and mammalian reactor-based protein-production technologies and the impending bottleneck in manufacturing capacity, plants are now emerging as an attractive alternative system with which to supply the growing need for protein-based therapeutics. However, full realization of the promise of plant-derived engineered proteins requires that we confront the dual challenges of bioequivalence and product consistency, challenges that are largely related to post-translational protein modifications (PTMs) that are crucial to the structure and function of most eukaryotic proteins. Among the protein PTMs, the foremost challenge for bioactivity and acceptance by the pharmaceutical and biotechnology industries and regulatory agencies is glycosylation. Advances made in recent years that 'humanize' plant glycosylation pathways combined with the discovery of terminal sialic acids (SAs) in plants now make feasible the bioengineering in plants of glycoproteins that have mammalian-like glycosylation. PMID- 15753006 TI - The clinical importance of the nomenclature, evolution and taxonomy of human papillomaviruses. AB - Human papillomaviruses (HPVs) are formally described by isolation of their circular double-stranded DNA genomes and establishment and comparison of the nucleotide sequence of these genomes. Alternatives such as serological diagnosis and maintenance of HPVs in culture are neither clinically useful nor consistently feasible. Novel HPV isolates have traditionally been described as "types". The analysis of specific HPV types is of medical importance, because HPV types typically induce type-specific lesions, i.e. they may be specific for cutaneous or mucosal epithelia, or give rise to benign warts or malignant carcinomas. Recently, it was formally decided that papillomaviruses are a virus family separate from the polyomaviruses. Within the papillomavirus family, closely or remotely related types form species or genera. These formal agreements were important as they brought the taxonomy of papillomaviruses in line with that of other viruses, bacteria and higher organisms, although their impact on medical practice and terminology used in clinical studies is limited. Notably, however, HPV types that are closely related (i.e. form "species") are associated with similar lesions. Confusion of the terms "type" and "subtype" should be avoided, as the latter term refers to some specific but rare taxonomic assemblages. In contrast to many RNA viruses, HPV types evolve very slowly, and diverged since the origin of humans only by about 2%. These divergent isolates are called "variants". HPVs evolved together with humankind and Homo sapiens was never without HPVs, and consequently never without warts and cervical cancer. Variants of the same HPV type may have different pathogenicity and may account for part of the worldwide disparities in the occurrence of genital cancers. PMID- 15753007 TI - The papillomavirus life cycle. AB - Papillomaviruses infect epithelial cells, and depend on epithelial differentiation for completion of their life cycle. The expression of viral gene products is closely regulated as the infected basal cell migrates towards the epithelial surface. Expression of E6 and E7 in the lower epithelial layers drives cells into S-phase, which creates an environment that is conducive for viral genome replication and cell proliferation. Genome amplification, which is necessary for the production of infectious virions, is prevented until the levels of viral replication proteins rise, and depends on the co-expression of several viral proteins. Virus capsid proteins are expressed in cells that also express E4 as the infected cell enters the upper epithelial layers. The timing of these events varies depending on the infecting papillomavirus, and in the case of the high-risk human papillomaviruses (HPVs), on the severity of neoplasia. Viruses that are evolutionarily related, such as HPV1 and canine oral papillomavirus (COPV), generally organize their productive cycle in a similar way, despite infecting different hosts and epithelial sites. In some instances, such as following HPV16 infection of the cervix or cottontail rabbit papillomavirus (CRPV) infection of domestic rabbits, papillomaviruses can undergo abortive infections in which the productive cycle of the virus is not completed. As with other DNA tumour viruses, such abortive infections can predispose to cancer. PMID- 15753008 TI - The epidemiology of human papillomavirus infections. AB - Infection with oncogenic human papillomavirus (HPV) types is a necessary cause of cervical cancer, the second most frequently occurring cancer in women worldwide. Rates of acquisition of HPV are high, particularly among sexually active young adults. Reported estimates of incident HPV infection among initially negative women have reached as high as 60% over a 5-year follow-up period. In this article, we review the epidemiology of HPV infection. In addition to estimates of disease frequency, we highlight risk factors for HPV infection, including the number of lifetime sex partners, which is the most salient risk factor. We discuss significant issues surrounding the natural history of HPV infection, including viral persistence versus clearance, immune response, development of lesions and development of cancer. Finally, we discuss strategies for preventing HPV infection. PMID- 15753009 TI - HPV-mediated transformation of the anogenital tract. AB - Infection with high-risk human papillomavirus (HR-HPV) has been associated with intraepithelial neoplasia and carcinomas at various sites of the anogenital tract, including the cervix, vulva, vagina, penis and anus. Although HR-HPV is a necessary cause for cervical cancer, the majority of anal cancers and a subset of cancers at other genital sites, additional (epi)genetic events are required for malignant transformation. HPV-mediated transformation of human epithelial cells has been recognized as a multi-step process resulting from deregulated transcription of the viral oncogenes E6 and E7 in the proliferating cells. Interference of E6 and E7 with cell cycle regulators induces genetic instability, which drives the continuous selection of oncogenic alterations providing cells with a malignant phenotype. Early genetic events during cervical carcinogenesis associated with immortalization, include deletions at chromosomes 3p, 6 and 10p, whereas amongst others gain of chromosome 3q, loss of chromosome 11 and epigenetic alterations such as inactivation of the TSLC1 tumor suppressor gene represent later events associated with tumor invasion. PMID- 15753010 TI - The role of human papillomavirus testing in cervical screening. AB - Organised, cytology-based cervical screening has led to a reduction in deaths associated with cervical cancer. Human papillomavirus (HPV) is necessary for the development of cervical cancer and associated pre-cursor cervical intraepithelial neoplasia and accumulated evidence suggests that incorporation of HPV testing could further refine screening programmes. HPV testing is discussed in the context of primary screening, for triage, and as a test of cure of treatment and possible value in developing countries. The high negative predictive value of a "double negative" cytology and HPV result could allow considerable changes in policy such as increased intervals between screening rounds, adjustment of age ranges for testing and schedule for return to routine screening post treatment. HPV testing for the triage of women to colposcopy with borderline or atypical squamous cells of undetermined significance (ASCUS) cytology could be clinically effective, but may be limited in women with low-grade squamous intraepithelial lesions (LSIL) or mild dyskaryosis by high HPV prevalence. Markers of HPV persistence harbour enormous potential to identify women at greatest risk of disease progression. Due to the diversity of existing cytology-based screening programmes, full cost-effectiveness analyses of HPV testing should be performed and assessed within local contexts. PMID- 15753011 TI - Molecular diagnosis of human papillomavirus (HPV) infections. AB - Human papillomaviruses (HPVs) comprise more than 100 genotypes. The mucosal types can be divided into high-risk and low-risk (LR) types depending on the associated disease risk. HPV infection is mainly diagnosed by molecular methods, since reliable serological tools are not available and culture of the virus is not possible. Accurate molecular diagnostic techniques that can be used to inform patient management and follow-up after treatment are now available for detection and identification of HPV. The diagnosis of HPV infections in patients at risk of disease in a clinical setting requires a different approach from that used for epidemiological studies, vaccination trials and natural history studies. This review describes the different molecular methods available for HPV detection and genotyping and their possible clinical utility. PMID- 15753013 TI - Human papillomavirus (HPV) in head and neck cancer. AB - The term "head and neck cancer" has been widely adopted in the recent literature, to include the lesions at several anatomic sites: the lip, oral cavity, nose and para-nasal sinuses, naso-pharynx, oro-pharynx, hypo-pharynx, and larynx. In this communication, the data on human papillomavirus (HPV) involvement in oral, oro pharyngeal, sino-nasal, and laryngeal carcinomas are reviewed. Our group was the first to present evidence on the involvement of HPV infections in both laryngeal and oral carcinogenesis, prompted by the discovery of morphological similarities between oral and cervical squamous cell lesions. The latest meta-analyses of the epidemiological studies as well as the multi-centre case-control studies have confirmed HPV as an independent risk factor for oral cancer, with a range of odds ratios (OR) between 3.7 and 5.4. Until 2002, 4768 oral carcinomas have been analysed for HPV DNA, and 22% were reported to contain HPV by any of the detection techniques. Of all non-genital cancers, tonsillar carcinomas appear to have the highest prevalence of HPV. By the end of 2002, 422 cases of tonsillar carcinoma have been analyzed for the presence of HPV DNA, with the overall detection rate of 51%. HPV 16 is the most prevalent HPV type found in 84% of HPV DNA-positive tumours. HPV seems to be mainly episomal in tonsillar carcinomas, but the significance of this observation is still obscure. Interestingly, patients with HPV 16-positive tumours seem to have a better overall- and disease specific survival, as compared with the HPV-negative group. To date, 1041 sino nasal papillomas have been analysed for HPV and 347 (33%) cases have been positive, whereas of the 322 sino-nasal carcinomas analysed so far, 70 (22%) have been positive for any HPV type. Laryngeal squamous cell papilloma and recurrent respiratory papillomatosis (RRP) are well-established HPV-induced tumours, whereas the role of HPV in laryngeal carcinomatosis remains controversial. The molecular mechanisms of HPV-associated carcinogenesis of the head and neck require further study. PMID- 15753012 TI - High-risk mucosal human papillomavirus infections during infancy & childhood. AB - Human papillomaviruses (HPVs) are small DNA tumour viruses associated with a variety of proliferative diseases. More than 100 types have been identified and can broadly be grouped into cutaneous and mucosal types according to their site of infection, and can be further subdivided into low-risk (LR) and high-risk (HR) types depending upon their association with malignancy. The main route of transmission of HR mucosal HPVs is through sexual contact, although the acquisition of virus cannot be entirely explained by this mode alone. Evidence also exists for horizontal transmission by other routes and vertical transmission. HR HPVs, particularly HPV-16, have been detected in oral swabs from newborns, infants and children. Such alternative modes of transmission and acquisition may have an important impact in several areas, including vaccination strategies, epidemiological studies, and the clinical management of children with HPV-associated diseases. This article reviews the literature describing the detection of HPV infections during infancy and childhood and provides evidence for a role of vertical transmission in the spread of HPV infection. PMID- 15753014 TI - Human papillomaviruses and skin cancer. AB - Cutaneous epidermodysplasia verruciformis (EV)-associated human papillomaviruses (HPVs) are found frequently in skin cancers especially in immunosuppressed people. They are also detectable in the normal skin and hair follicles of a proportion of individuals who have no immune defect. The available evidence to link HPVs causally with skin carcinogenesis is not conclusive, but includes epidemiological, molecular and immunological studies. PMID- 15753015 TI - Immune control of human papillomavirus (HPV) associated anogenital disease and potential for vaccination. AB - This review discusses: (1) immune mechanisms relevant to the natural control of a human papillomavirus (HPV) infection; (2) viral strategies to evade or subvert immune attack; (3) the significance of immune escape as a feature of the evolution of invasive cancer; (4) vaccine strategies for prevention and/or therapy. HPV infection and associated malignancy can induce humoral and cellular immunity to capsid and oncogene viral proteins, but it is not always clear whether such responses are a consequence of the disease rather than the resolving factor(s). Prophylactic strategies are utilising virus-like particles (VLP) composed of the L1 viral capsid protein to induce neutralising antibodies, while therapeutic approaches are aimed at generating specific T cells targeted at the viral E6 and/or E7 oncogenes. Thus far, HPV VLP vaccines have proved clinically efficacious in the early clinical trials to prevent infection. Different types of therapeutic vaccines including peptide, protein, DNA or viral vector encoded have proved safe and immunogenic in patients, although there is often no correlation with clinical outcome. Understanding the equilibrium between viral and immunological factors will be important in providing the appropriate tools to evoke effective prophylactic and therapeutic immunity. It seems likely that combined prophylactic and therapeutic vaccine approaches could offer the best prospect for any significant reduction in death from cervical cancer worldwide. PMID- 15753016 TI - Therapy for genital human papillomavirus-related disease. AB - Genital human papillomavirus (HPV) infection is very common, and often sub clinical and usually resolves without any treatment. Genital warts are caused by HPV 6/11 infection and are one of the commonest clinically recognised disease manifestations of genital HPV. Subjects with genital warts usually perceive them as cosmetically disfiguring, often leading to adverse psychological symptoms, and most subjects with genital warts will present requesting treatment. A wide variety of treatments are available including both provider- and patient-applied therapies. Various individual subject and disease factors mediate appropriate therapy choice. Some of the treatments that are used for genital warts can also be used for some cases of intraepithelial neoplasia caused by high-oncogenic risk HPVs occurring at vulval, anal or penile sites. Specific treatment considerations apply to genital warts in pregnancy and laryngeal papillomatosis and these are also discussed. PMID- 15753017 TI - A new licence to clone. AB - Many countries are still grappling with the issue of therapeutic cloning of human cells as the UN tackles this issue alongside reproductive cloning, but in the UK a new programme looking at the causes of motor neuron disease has recently been sanctioned. Michael Gross reports. PMID- 15753019 TI - Planet of the apes. AB - What makes us humans so special? Our language, our genes, our culture, our cognitive skills? At the Max-Planck-Institute for Evolutionary Anthropology in Leipzig, psychologists, linguists and biologists tackle this old question in a truly multidisciplinary way. Their results have implications not just for our understanding of human evolution--they also touch directly on many social and environmental issues. Florian Maderspacher reports. PMID- 15753020 TI - Q & A. [interview]. AB - Elaine and Gary Ostrander spent their youth in New Jersey and New York before heading to Nebraska for their teen years and eventually Washington State for High School and college, as their father moved around in library administration. Elaine was an undergraduate at the University of Washington, a graduate student at the Oregon Health Sciences University and a postdoc with James Wang at Harvard, studying DNA supercoiling. She next went to Berkeley, where she began the canine genome project, initiating the meiotic linkage map and working on human chromosome 21 at the Lawrence Berkeley National Labs. In 1993 she moved to the Fred Hutchinson Cancer Research Center where she is now a Member of the Divisions of Clinical Research and Human Biology. She is also an Affiliate Professor of Genome Sciences and Biology at the University of Washington, and heads the Program in Genetics at the Hutchinson Center. Gary completed his undergraduate degree in Biology at Seattle University, a M.S. degree at Illinois State University and a Ph.D at the University of Washington in Ocean and Fisheries Science. He went on to be a postdoc in the Department of Pathology at the University of Washington Medical School while being mentored by Senitroh Hakomori of the Fred Hutchinson Cancer Research Center and Eric Holmes of the Pacific Northwest Research Foundation. His work focused on using novel aspects of the biology of fishes to address fundamental questions about cancer. He subsequently held both faculty and administrative positions at Oklahoma State University. Since 1996, he has been at the Johns Hopkins University, where he currently holds academic appointments in the Departments of Biology and Comparative Medicine and is the Associate Provost for Research. PMID- 15753021 TI - Major sperm protein. PMID- 15753022 TI - Neurotransmitters. PMID- 15753023 TI - Geography predicts neutral genetic diversity of human populations. PMID- 15753024 TI - Magnetosensation in zebrafish. PMID- 15753025 TI - Evolutionary ecology: wasp mother's little helpers. AB - The medical application of antibiotics dramatically reduced human infant mortality in the previous century. A new study indicates that ground nesting wasps exploit Streptomyces strains that they rear in their antennae for the same purpose. PMID- 15753026 TI - Protein localization: reach out and touch the forespore. AB - Bacterial proteins are typically sorted to subcellular regions with distinct physical characteristics that serve as cellular 'addresses', but many proteins are evidently sorted to specific areas that lack any apparent unique identity. Recent work in Bacillus subtilis suggests that such proteins may be localized by interacting with extracellular domains of proteins in an adjacent cellular compartment. PMID- 15753027 TI - Virology: Gulliver among the Lilliputians. AB - The discovery and genome sequencing of the mimivirus, a parasite of Acanthamoeba, blurs the boundary between viruses and cells: the 1.2 Mb genome of the mimivirus is predicted to contain 1262 genes and is much bigger than the genomes of many parasitic bacteria. PMID- 15753028 TI - Polytopic proteins: preventing aggregation in the membrane. AB - It has been proposed that the aggregation of nascent transmembrane segments of polytopic proteins is prevented by chaperones present in the endoplasmic reticulum membrane; now the first experimental support for this proposal has been reported. PMID- 15753029 TI - Ant navigation: reading geometrical signposts. AB - Ants often travel along complex pheromone trail systems between their nest and foraging areas. A new and surprising discovery is that Pharaoh's ants can work out from the geometry of individual branch points on the trail whether they are heading towards or away from the nest. PMID- 15753030 TI - Evolution of cooperation: does selfishness restraint lie within? AB - Traditional models of how cooperative strategies succeed in evolution have largely focused on social interactions among individuals and selection acting at kin and group levels. A recent study at the genetic level suggests that cooperation may also be promoted by the evolution of gene-trait relationships that limit the range of possible cheating mechanisms that can evolve. PMID- 15753031 TI - Hedgehog signaling: an Arrestin connection? AB - Arrestins are best known for terminating signaling by G protein coupled receptors. New binding, localization and genetic studies suggest that Arrestins may also participate in the transduction of Hedgehog signals by the seven transmembrane domain protein, Smoothened. PMID- 15753032 TI - Mitosis: PARty time in the spindle. AB - Poly(ADP-ribose), a post-translational protein modification known to affect chromatin structure, has now been shown to regulate microtubule organization during mitosis. These findings alter conventional views of the mechanisms of spindle assembly and function. PMID- 15753033 TI - The KLP-6 kinesin is required for male mating behaviors and polycystin localization in Caenorhabditis elegans. AB - BACKGROUND: Male mating behavior of the nematode Caenorhabditis elegans offers an intriguing model to study the genetics of sensory behavior, cilia function, and autosomal dominant polycystic kidney disease (ADPKD). The C. elegans polycystins LOV-1 and PKD-2 act in male-specific sensory cilia required for response and vulva-location mating behaviors. RESULTS: Here, we identify and characterize a new mating mutant, sy511. sy511 behavioral phenotypes were mapped to a mutation in the klp-6 locus, a gene encoding a member of the kinesin-3 family (previously known as the UNC-104/Kif1A family). KLP-6 has a single homolog of unknown function in vertebrate genomes, including fish, chicken, mouse, rat, and human. We show that KLP-6 expresses exclusively in sensory neurons with exposed ciliated endings and colocalizes with the polycystins in cilia of male-specific neurons. Cilia of klp-6 mutants appear normal, suggesting a defect in sensory neuron function but not development. KLP-6 structure-function analysis reveals that the putative cargo binding domain directs the motor to cilia. Consistent with a motor cargo association between KLP-6 and the polycystins, klp-6 is required for PKD-2 localization and function within cilia. Genetically, we find klp-6 regulates behavior through polycystin-dependent and -independent pathways. CONCLUSION: Multiple ciliary transport pathways dependent on kinesin-II, OSM-3, and KLP-6 may act sequentially to build cilia and localize sensory ciliary membrane proteins such as the polycystins. We propose that KLP-6 and the polycystins function as an evolutionarily conserved ciliary unit. KLP-6 promises new routes to understanding cilia function, behavior, and ADPKD. PMID- 15753034 TI - Association of CNK1 with Rho guanine nucleotide exchange factors controls signaling specificity downstream of Rho. AB - BACKGROUND: Rho is a small GTPase that controls signal transduction pathways in response to a large number of extracellular stimuli. With over 15 potential Rho target proteins identified to date, however, it is not clear how distinct signaling outputs can be generated downstream of a particular stimulus. RESULTS: Several of the known Rho targets are structurally reminiscent of scaffold proteins, which are generally thought to play an important role in controlling signaling specificity. Here, we show that the Rho target CNK1 is a scaffold protein that interacts with Net1 or p115RhoGEF, two Rho-specific guanine nucleotide exchange factors (GEFs), as well with MLK2 and MKK7, two of the kinase components in the JNK MAP kinase cascade. CNK1 acts cooperatively with the two GEFs to activate JNK MAP kinase, but not other Rho-mediated pathways. In HeLa cells, serum or sphingosine-1-phosphate stimulate Rho-dependent activation of the JNK MAP kinase cascade, and this requires endogenous CNK1. CONCLUSIONS: We conclude that CNK1 couples a subset of Rho exchange factors to activation of the JNK MAP kinase pathway and that signaling specificity is achieved through complexes containing both upstream activators and downstream targets of Rho. PMID- 15753035 TI - Fusogenic activity of EFF-1 is regulated via dynamic localization in fusing somatic cells of C. elegans. AB - BACKGROUND: Many animal tissues form via fusion of cells. Yet in all instances of developmental cell fusion, the mechanism underlying fusion of plasma membranes remains poorly understood. EFF-1 is required for most somatic cell fusions in C. elegans, and misexpressed EFF-1 alters the normal pattern of fusing hypodermal cells. However, the autonomous activity of EFF-1, the rules governing its specificity, and the mechanism of its action have not been examined. RESULTS: We show that EFF-1 acts as a cellular fusogen, capable of inducing fusion of virtually any somatic cells in C. elegans, yet targeted precisely to fusion-fated contacts during normal development. Misexpression of EFF-1 in early embryos causes fusion among groups of cells composed entirely of nonfusion-fated members. Measurements of cytoplasm diffusion in induced fusion events show that ectopic EFF-1 expression produces fusion pores similar to those in normal fusion events. GFP-labeled EFF-1 is specifically targeted to fusion-competent cell contacts via reciprocal localization to the touching membranes of EFF-1-expressing cells. EFF 1 function is also governed by intercellular barriers that prohibit cell fusion between distinct tissues. Analysis of mutant versions of EFF-1 indicates a novel mode of fusogenicity, employing neither a phospholipase active site nor hydrophobic fusion-peptide acting solely in pore formation. CONCLUSIONS: EFF-1 can confer potent fusogenic activity to nonfusing cell types. However, it is normally targeted only to fusion-fated cell borders via mutual interaction between EFF-1-expressing cells and relocalization to the plasma membrane. Because EFF-1 appears evolutionarily unique to nematodes, multiple mechanisms may have evolved for controlled plasma-membrane fusion in development. PMID- 15753036 TI - Asymmetrical activation in the human brain during processing of fearful faces. AB - Traditional split-field studies and patient research indicate a privileged role for the right hemisphere in emotional processing [1-7], but there has been little direct fMRI evidence for this, despite many studies on emotional-face processing [8-10](see Supplemental Background). With fMRI, we addressed differential hemispheric processing of fearful versus neutral faces by presenting subjects with faces bilaterally [11-13]and orthogonally manipulating whether each hemifield showed a fearful or neutral expression prior to presentation of a checkerboard target. Target discrimination in the left visual field was more accurate after a fearful face was presented there. Event-related fMRI showed right-lateralized brain activations for fearful minus neutral left-hemifield faces in right visual areas, as well as more activity in the right than in the left amygdala. These activations occurred regardless of the type of right hemifield face shown concurrently, concordant with the behavioral effect. No analogous behavioral or fMRI effects were observed for fearful faces in the right visual field (left hemisphere). The amygdala showed enhanced functional coupling with right-middle and anterior-fusiform areas in the context of a left-hemifield fearful face. These data provide behavioral and fMRI evidence for right lateralized emotional processing during bilateral stimulation involving enhanced coupling of the amygdala and right-hemispheric extrastriate cortex. PMID- 15753037 TI - Real-time imaging of DNA ejection from single phage particles. AB - Infection by tailed dsDNA phages is initiated by release of the viral DNA from the capsid and its polarized injection into the host. The driving force for the genome transport remains poorly defined. Among many hypothesis [1], it has been proposed that the internal pressure built up during packaging of the DNA in the capsid is responsible for its injection [2-4]. Whether the energy stored during packaging is sufficient to cause full DNA ejection or only to initiate the process was tested on phage T5 whose DNA (121,400 bp) can be released in vitro by mere interaction of the phage with its E. coli membrane receptor FhuA [5-7]. We present a fluorescence microscopy study investigating in real time the dynamics of DNA ejection from single T5 phages adsorbed onto a microfluidic cell. The ejected DNA was fluorescently stained, and its length was measured at different stages of the ejection after being stretched in a hydrodynamic flow. We conclude that DNA release is not an all-or-none process but occurs in a stepwise fashion and at a rate reaching 75,000 bp/sec. The relevance of this stepwise ejection to the in vivo DNA transfer is discussed. PMID- 15753038 TI - Requirement of homeobox gene STIMPY/WOX9 for Arabidopsis meristem growth and maintenance. AB - Most organs of flowering plants develop postembryonically from groups of pluripotent cells called meristems [1]. The shoot apical meristem (SAM) is specified by two complementary pathways [2-4]. SHOOT MERISTEMLESS (STM; [5]) defines the entire SAM region [6]. WUSCHEL (WUS), on the other hand, functions in a more restricted set of cells to promote stem-cell fate and is regulated by the CLAVATA genes in a negative feedback loop [7-10]. In contrast, little is known about how the growth of the SAM, which increases in size during vegetative development [11], is regulated. We have characterized STIMPY (STIP; also called WOX9 [12]), a homeobox gene required for the growth of the vegetative SAM, in part by positively regulating WUS expression. In addition, STIP is required in several other aerial organs and the root. What sets STIP apart from STM and WUS is that stip mutants can be fully rescued by stimulating the entry into the cell cycle with sucrose. Therefore, STIP is likely to act in all these tissues by maintaining cell division and preventing premature differentiation. Taken together, our findings suggest that STIP identifies a new genetic pathway integrating developmental signals with cell-cycle control. PMID- 15753039 TI - Rhesus monkeys attribute perceptions to others. AB - Paramount among human cognitive abilities is the capacity to reason about what others think, want, and see--a capacity referred to as a theory of mind (ToM). Despite its importance in human cognition, the extent to which other primates share human ToM capacities has for decades remained a mystery. To date, primates [1, 2] have performed poorly in behavioral tasks that require ToM abilities, despite the fact that some macaques are known to encode social stimuli at the level of single neurons [3-5]. Here, we presented rhesus macaques with a more ecologically relevant ToM task in which subjects could "steal" a contested grape from one of two human competitors. In six experiments, monkeys selectively retrieved the grape from an experimenter who was incapable of seeing the grape rather than an experimenter who was visually aware. These results suggest that rhesus macaques possess an essential component of ToM: the ability to deduce what others perceive on the basis of where they are looking. These results converge with new findings illustrating the importance of competitive paradigms in apes [6]. Moreover, they raise the possibility that, in primates, cortical cells thought to encode where others are looking [7] may encode what those individuals see as well. PMID- 15753040 TI - Perceiving time to collision activates the sensorimotor cortex. AB - The survival of many animals hinges upon their ability to avoid collisions with other animals or objects, or to precisely control the timing of collisions. Optical expansion provides a compelling impression of object approach and in principle can provide the basis for judgments of time to collision (TTC) [1]. It has been demonstrated that pigeons [2] and houseflies [3] have neural systems that can initiate rapid coordinated actions on the basis of optical expansion. In the case of humans, the linkage between judgments of TTC and coordinated action has not been established at a cortical level. Using functional magnetic resonance imaging (fMRI), we identified superior-parietal and motor-cortex areas that are selectively active during perceptual TTC judgments, some of which are normally involved in producing reach-to-grasp responses. These activations could not be attributed to actual movement of participants. We demonstrate that networks involved in the computational problem of extracting TTC from expansion information have close correspondence with the sensorimotor systems that would be involved in preparing a timed motor response, such as catching a ball or avoiding collision. PMID- 15753041 TI - Regulation of bidirectional melanosome transport by organelle bound MAP kinase. AB - Regulation of intracellular transport plays a role in a number of processes, including mitosis, determination of cell polarity, and neuronal growth. In Xenopus melanophores, transport of melanosomes toward the cell center is triggered by melatonin, whereas their dispersion throughout the cytoplasm is triggered by melanocyte-stimulating hormone (MSH), with both of these processes mediated by cAMP-dependent protein kinase A (PKA) activity [1, 2]. Recently, the ERK (extracellular signal-regulated kinase) pathway has been implicated in regulating organelle transport and signaling downstream of melatonin receptor [3, 4]. Here, we directly demonstrate that melanosome transport is regulated by ERK signaling. Inhibition of ERK signaling by the MEK (MAPK/ERK kinase) inhibitor U0126 blocks bidirectional melanosome transport along microtubules, and stimulation of ERK by constitutively active MEK1/2 stimulates transport. These effects are specific because perturbation of ERK signaling has no effect on the movement of lysosomes, organelles related to melanosomes [5]. Biochemical analysis demonstrates that MEK and ERK are present on melanosomes and transiently activated by melatonin. Furthermore, this activation correlates with an increase in melanosome transport. Finally, direct inhibition of PKA transiently activates ERK, demonstrating that ERK acts downstream of PKA. We propose that signaling of organelle bound ERK is a key pathway that regulates bidirectional, microtubule based melanosome transport. PMID- 15753042 TI - Mitotic spindles and cleavage planes are oriented randomly in the two-cell mouse embryo. AB - Most experimental embryological studies performed on the early mouse embryo have led to the conclusion that there are no mosaically distributed developmental determinants in the zygote and early embryo (for example see [1-6]). It has been suggested recently that "the cleavage pattern of the early mouse embryo is not random and that the three-dimensional body plan is pre-patterned in the egg" (in [7] for review see [8-10]). Two major spatial cues influencing the pattern of cleavage divisions have been proposed: the site of the second meiotic division [11, 12] and the sperm entry point [13-14], although the latter is controversial [15-17]. An implication of this hypothesis is that the orientations of the first few cleavage divisions are stereotyped. Such a define cleavage pattern, leading to the segregation of developmental determinants, is observed in many species [18]. Recently, it was shown that the first cleavage plane is not predetermined but defined by the topology of the two apposing pronuclei [19]. Because the position of the female pronucleus is dependent upon the site of polar body extrusion and the position of the male pronuclei is dependent upon the sperm entry point [19-20], this observation leaves open the possibility that the sperm may provide some kind of directionality [7]. But, even if asymmetries were set up only after fertilization, a stereotyped cleavage pattern should take place during the following cleavage divisions. Thus, we studied the cleavage pattern of two cell embryos by videomicroscopy to distinguish between the two hypotheses. After the mitotic spindle formed, its orientation did not change until cleavage. During late metaphase and anaphase, the spindle poles appear to be anchored to the cortex through astral microtubules and PARD6a. Only at the time of cleavage, during late anaphase, do the forming daughter cells change their relative positions. These studies show that cleavage planes are oriented randomly in two cell embryos. This argues against a prepatterning of the mouse embryo before compaction. PMID- 15753043 TI - MutSalpha binds to and promotes synapsis of transcriptionally activated immunoglobulin switch regions. AB - Immunoglobulin class switch recombination joins a new constant (C) region to the rearranged and expressed heavy chain variable (VDJ) region in antigen-activated B cells (Figure 1A) (reviewed in [1, 2]). Switch recombination is activated by transcription of intronic, G-rich and repetitive switch (S) regions and produces junctions that are heterogeneous in sequence and position in the S regions. Switch recombination depends upon the B cell-specific cytidine deaminase, AID, and conserved DNA repair factors, including the mismatch repair heterodimer, MutSalpha (MSH2/MSH6). In mice, ablation of Msh2 or Msh6, but not Msh3, decreases levels of switch recombination and diminishes heterogeneity of switch junctions [3-7]. Here, we demonstrate that MSH2 associates with transcribed S regions in primary murine B cells activated for switch recombination. Electron microscopic imaging reveals that MutSalpha binds in vitro to DNA structures formed within transcribed S regions and mediates their synapsis. MutSalpha binds with high affinity to G4 DNA formed upon transcription of the S regions and also binds to U.G mismatches, initial products of DNA deamination by AID. These results suggest that MutSalpha interacts with the S regions in switching B cells to promote DNA synapsis and recombination. PMID- 15753044 TI - Symbiotic bacteria protect wasp larvae from fungal infestation. AB - Symbiotic associations between different organisms are of great importance for evolutionary and ecological processes [1-4]. Bacteria are particularly valuable symbiotic partners owing to their huge diversity of biochemical pathways that may open entirely new ecological niches for higher organisms [1-3]. Here, we report on a unique association between a new Streptomyces species and a solitary hunting wasp, the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae). Beewolf females cultivate the Streptomyces bacteria in specialized antennal glands and apply them to the brood cell prior to oviposition. The bacteria are taken up by the larva and occur on the walls of the cocoon. Bioassays indicate that the streptomycetes protect the cocoon from fungal infestation and significantly enhance the survival probability of the larva, possibly by producing antibiotics. Behavioral observations strongly suggest a vertical transmission of the bacteria. Two congeneric beewolf species harbor closely related streptomycetes in their antennae, indicating that the association with protective bacteria is widespread among philanthine wasps and might play an important role in other insects as well. This is the first report on the cultivation of bacteria in insect antennae and the first case of a symbiosis involving bacteria of the important antibiotic-producing genus Streptomyces. PMID- 15753045 TI - The zebrafish-secreted matrix protein you/scube2 is implicated in long-range regulation of hedgehog signaling. AB - The Hedgehog (Hh) signal plays a pivotal role in induction of ventral neuronal and muscle cell types around the midline during vertebrate development [1]. We report that the gene disrupted in zebrafish you mutants, in which Hh signaling is impaired, encodes the secreted matrix protein Scube2. Consistently, epistasis analyses suggested that Scube2 functions upstream of Hh ligands or through a parallel pathway. In addition, overexpression analyses suggested that Scube2 is an essential, but a permissive, mediator of Hh signaling in zebrafish embryos. Surprisingly, the you gene is expressed in the dorsal neural tube, raising the possibility that Scube2 could indirectly act via a long-range regulator of Hh signaling. The dorsal Bmps have a long-range and opposing influence on Hh signaling [2-5]. We show that neural plate patterning is affected in you mutants in a way that is consistent with the aberrant long-range action of a Bmp dependent signal. We further show that Bmp activity can be attenuated by the coexpression of Scube2. Our data support the idea that Scube2 can modulate the long-range action of Bmp-dependent signaling in the neural tube and somites. PMID- 15753046 TI - Linking brainwaves to the brain: an ERP primer. AB - This article reviews literature on the characteristics and possible interpretations of the event-related potential (ERP) peaks commonly identified in research. The description of each peak includes typical latencies, cortical distributions, and possible brain sources of observed activity as well as the evoking paradigms and underlying psychological processes. The review is intended to serve as a tutorial for general readers interested in neuropsychological research and as a reference source for researchers using ERP techniques. PMID- 15753047 TI - Length effect in word naming in reading: role of reading experience and reading deficit in italian readers. AB - Vocal reaction times (RTs) in naming 3- to 8-letter words were measured in proficient and dyslexic readers (Study 1). In proficient readers, RTs were independent of word length up to 5-letter words, indicating parallel processing. In the 5- to 8-letter range, RTs increased linearly, indicating sequential processing. Reading experience was associated with both faster discrimination of individual elements and parallel processing of increasingly large word parts. In dyslexics, RTs increased linearly with increasing length indicating reliance on sequential decoding. Individual analysis indicated 2 profiles of RTs (Types A and B). In Study 2, the distinction between A and B dyslexics was not associated with the use of different reading procedures. However, a more marked speed deficit characterized Type B dyslexics. PMID- 15753049 TI - Reaching in children with and without developmental coordination disorder under normal and perturbed vision. AB - The aim of this study is to describe the reaching action of children with developmental coordination disorder (DCD) and to investigate whether their use of visual feedback during the time course of this action differs from that of normally developing children. Fifty-two children subdivided into 2 age bands (7-8 and 9-10 years) within 2 groups (with and without DCD) participated in this experiment. They were asked to reach for a target positioned either ipsilaterally or contralaterally to the reaching hand in 2 visual conditions: a condition where vision was unrestrained (normal vision) and a condition where they wore glasses with prismatic lenses (perturbed vision). An analysis of the experimental data indicates that the trajectories followed by the DCD group were longer and more curved than those of the control group. Further, the deceleration times were longer for the DCD group than for the normally developing children. The introduction of the prismatic lenses supports the idea that the use of visual feedback by children with DCD may be different from that in children without DCD. PMID- 15753048 TI - The development of endogenous orienting: control over the scope of attention and lateral asymmetries. AB - This study examined the development of endogenous orienting in children ages 6, 10, and 14 years and in adults. Participants were asked to respond with a button press to targets appearing in the left or right visual field. Cues that correctly or incorrectly indicated the target's location preceded the appearance of targets at stimulus onset asynchronies (SOAs) of 100 and 800 msec. Cues that signaled a target's appearance, but not its location, were also included. In addition to raw reaction times, orienting effects, benefits, and costs were examined. Two main findings emerged: First, for all participants except the youngest children effect sizes increased with increasing SOA. The absence of an SOA effect in the youngest group is interpreted as evidence of their difficulty in voluntarily adjusting the scope of their attentional focus. In addition, field asymmetries changed with the age of the respondent: 6-year-olds showed a right-field advantage, 10-year-olds failed to show any laterality differences, and 14-year-olds and adults responded more quickly to targets in the left than in the right field. This finding is consistent with developmental data on a number of cognitive processes, and is interpreted within a developmental framework of right-hemisphere dominance for spatial orienting (cf. Mesulam, 1998). PMID- 15753050 TI - Cognitive heterogeneity in Williams syndrome. AB - This study used the Woodcock-Johnson Tests of Cognitive Ability-Revised to investigate a wide range of cognitive abilities in people with Williams syndrome (WS). It involved a comparatively large sample of 31 people with WS, but took a case-series approach. The study addressed the widespread claims of a characteristic "WS cognitive profile" by looking for heterogeneity rather than homogeneity. People with WS showed a variety of preserved (significantly above mental age [MA]), expected (at MA), and significantly impaired (significantly below MA) levels of functioning. Such results provide clear evidence for heterogeneity in cognitive functions within WS. We found the most homogeneity on a test of phonological processing and a test of phonological short-term memory, with half of the WS sample performing at MA levels on these tests. Interestingly, no WS individual showed a weakness on a test of nonverbal reasoning, and only one WS individual showed a weakness on a test of verbal comprehension. In addition, we found that strengths on analysis-synthesis and verbal analogies occurred only for WS individuals with an MA less than 5.5 years (our sample median MA); people with an MA greater than 5.5 years performed at MA level on these 2 tests. Results also provided preliminary evidence for distinct subgroups of WS people based on their cognitive strengths and weaknesses on a broad range of cognitive functions. On the basis of the findings, caution should be made in declaring a single cognitive profile that is characteristic of all individuals with WS. Just as there is heterogeneity in genetic and physical anomalies within WS, not all WS individuals share the same cognitive strengths and weaknesses. Also, not all WS individuals show the profile of a strength in verbal abilities and a weakness in spatial functions. PMID- 15753051 TI - Acute lung Injury in a healthy donor during mobilization of peripheral blood stem cells using granulocyte-colony stimulating factor alone. AB - Granulocyte-colony stimulating factor (G-CSF), a hematopoietic growth factor, is widely used to accelerate recovery from neutropenia after severe chemotherapy, both decreasing the risk of infection and mobilizing peripheral blood stem cells. Adverse effects occur with G-CSF use in approximately 30% of cases, comprised predominantly of bone pain, headache, and general fatigue. Pulmonary toxicity is very rare. Here, we describe a healthy donor for allogeneic hematopoietic stem cell transplantation who developed acute lung injury (ALI) after 4 days of G-CSF administration. Among the serum cytokines examined, only Interleukin (IL)-1beta level was elevated in this case. As a high level of IL-1beta was detected at the onset of ALI, on day 4 after G-CSF administration, and decreased to below the level of detection on day 11, it is possible in a certain part that IL-1beta was involved in the onset of G-CSF-related ALI in the present case. Granulocyte colony stimulating factor (G-CSF) is commonly administered to healthy donors to mobilize peripheral blood stem cells (PBSC) for allogeneic hematopoietic stem cell transplantation (allo-HSCT). Adverse events from G-CSF use in healthy donors have been described in approximately 30% of cases, and are comprised predominantly of bone pain, headache, and general fatigue. Pulmonary complications caused by G-CSF include cough, dyspnea, and interstitial or alveolar pulmonary edema with mild-to-severe deterioration of blood oxygen level. Few cases of acute respiratory distress syndrome (ARDS) following G-CSF administration have been reported. The present report describes a healthy donor for allo-HSCT with acute lung injury (ALI) after 4 days of G-CSF administration. The cytokine-related mechanisms of G-CSF administration that contribute to ALI are discussed. PMID- 15753052 TI - Cyclosporin neurotoxicity with Epstein-Barr virus-associated hemophagocytic syndrome. AB - In September 2000, a 22-year-old female was admitted to our hospital due to high grade fever, liver enzymes elevation and pancytopenia. Bone marrow aspiration was performed, and hemophagocytosis was present. Epstein-Barr virus (EBV) DNA was positive in her peripheral blood, and we diagnosed the case as EBV-associated hemophagocytic syndrome (EB-VAHS) after excluding other malignancies. The initial therapy including etoposide and dexamethasone was started. As severe leukocytopenia developed, etoposide was stopped and cyclosporin A (CsA) was administered continuously. Four days after administration of CsA, she developed convulsive seizures with loss of consciousness. An MRI demonstrated decreased signal with T1-weighting and high signal with T2-weighting in the subcortical white matter including the posterior lobe. We stopped CsA infusion, and glycerol was administered. Soon the symptom disappeared. When patients developed an episode of convulsive seizure, other diagnostic possibilities were central nervous system (CNS) involvement of hemophagocytosis, EBV encephalitis and acute disseminated encephalomyelitis (ADEM). CsA neurotoxicity must be considered even in the case of EB-VAHS with administration of CsA. As previously reported, Fluid attenuated Inversion Recovery (FLAIR) imaging improved diagnostic confidence and conspicuity of the T2 hyper intense lesions of CsA neurotoxicity, as well as tacrolimus encephalopathy, typically in the subcortical white matter. Key words; Cyclosporin neurotoxicity; Epstein-Barr virus associated-Hemophagocytic syndrome; Magnetic Resonance Image (MRI). PMID- 15753053 TI - Autoimmune type antiphospholipid antibodies in a patient with Q fever. PMID- 15753054 TI - HTLV-1 associated adult T-cell leukemia/lymphoma in Israel: report of two patients of Romanian origin. AB - Human T-lymphotropic virus type 1 (HTLV-1) was the first human oncovirus isolated by Gallo et al. in 1980 and established as an etiological agent for adult T-cell leukemia/ lymphoma (ATL). Although more than 15 million individuals are infected by HTLV-1 through the world, the spread of the virus is highly endemic. The HTLV 1 infection is prevailing in southwestern Japan, inter-tropical Africa, Central and South America. In Kyushu district, Japan, the seroprevalence reaches >30% in the adult population. In the US, Europe and the Middle East the HTLV-1 infection is very rare, and cases of ATL have been reported sporadically. We describe here acute ATL in two patients of Jewish- Romanian origin. The epidemiological anamnesis and screening indicate that both patients acquired the HTLV-1 from their mothers leaving in Romania. PMID- 15753055 TI - Pentostatin (2'-deoxycoformycin) for the treatment of hepatosplenic gammadelta T cell lymphomas. AB - We report the results of treatment with single agent 2'-deoxycoformycin (Pentostatin) in two patients with Hepatosplenic gammadelta T-cell lymphoma (HSgammadeltaTCL), a rare lymphoma subtype with a highly unfavorable prognosis. Present and previous data reviewed here demonstrates the striking cytotoxic activity of Pentostatin against gammadelta+ tumor T cells. Further studies are warranted to define the optimal strategy to fully exploit therapeutic potential of this drug in patients with HSgammadeltaTCL. PMID- 15753056 TI - Deep lamellar keratoplasty for corneal perforation due to chronic graft-versus host disease following allogeneic hematopoietic stem-cell transplantation. PMID- 15753057 TI - Clinical heterogeneity of acquired hemophilia A: a description of 4 cases. AB - Acquired hemophilia A is a rare but severe auto-immune bleeding disorder characterized by the presence of autoantibodies directed against clotting factor VIII. Acquired hemophilia A may be idiopathic or associated with several conditions, such as postpartum, autoimmune diseases, malignancies or drugs. The treatment modalities of bleeding episodes and eradication of the factor VIII auto antibody depend on the titer of anti-factor VIII:C and may include desmopressin (DDAVP), prednisolone, prednisolone-cyclophosphamide, high dose intravenous gammaglobulin, FVIII-VWF concentrate and/or recombinant FVIIa (rFVIIa). In this study we report four cases of autoimmune factor VIII inhibitors (2 associated with autoimmune disorders, 2 idiopathic) demonstrating the heterogeneity of this disease from pathogenic, clinical, therapeutic and prognostic points of view. PMID- 15753058 TI - Catastrophic antiphospholipid syndrome: atypical presentation in the setting of chronic graft versus host disease: case report and review of the literature. AB - A 44-year-old female developed a catastrophic thrombotic syndrome following allogeneic stem cell transplantation for acute lymphoblastic leukemia. In the context of chronic graft versus host disease, she developed a lethal multi-system thrombotic state as evidenced by stroke, renal failure and cardiac thrombi in association with elevated anticardiolipin antibody. The case is discussed in the framework of the existing literature and derives clinical practice recommendations for this rare but clinically devastating entity. PMID- 15753059 TI - Imatinib mesylate (STI-571) and porphyria cutanea tarda in a Chinese patient. PMID- 15753060 TI - Successful treatment with allogeneic bone marrow transplantation of an early relapse of ALK-positive anaplastic large cell lymphoma. PMID- 15753061 TI - Cortical cell assemblies: a possible mechanism for motor programs. AB - The concept of a motor program has been used to interpret a diverse range of empirical findings related to preparation and initiation of voluntary movement. In the absence of an underlying mechanism, its exploratory power has been limited to that of an analogy with running a stored computer program. We argue that the theory of cortical cell assemblies suggests a possible neural mechanism for motor programming. According to this view, a motor program may be conceptualized as a cell assembly, which is stored in the form of strengthened synaptic connections between cortical pyramidal neurons. These connections determine which combinations of corticospinal neurons are activated when the cell assembly is ignited. The dynamics of cell assembly ignition are considered in relation to the problem of serial order. These considerations lead to a plausible neural mechanism for the programming of movements and movement sequences that is compatible with the effects of precue information and sequence length on reaction times. Anatomical and physiological guidelines for future quantitative models of cortical cell assemblies are suggested. By taking into account the parallel re entrant loops between the cerebral cortex and basal ganglia, the theory of cortical cell assemblies suggests a mechanism for motor plans that involve longer sequences. The suggested model is compared with other existing neural network models for motor programming. PMID- 15753062 TI - Quantitative analysis of human movement synergies: constructive pattern analysis for gait. AB - To record three-dimensional coordinates of the joints from normal human subjects during locomotion, we used a digital motion analysis system (ELITE). Recordings were obtained under several different conditions, which included normal walking and stepping over obstacles. Principal component analysis was used to analyze coordinate data after conversion of the data to segmental angles. This technique gave a stable summary of the redundancy in gait kinematic data in the form of reduced variables (principal components). By modeling the shapes of the phase plots of reduced variables (distortion analysis) and using a limited number of model parameters, good resolution was obtained between subtly different conditions. Hence, it was possible to accurately resolve small distributed changes in gait patterns within subjects. These methods seem particularly suited to longitudinal studies in which relevant movement features are not known a priori. Assumptions and neurophysiological applications are discussed. PMID- 15753063 TI - A statistical approach to sensorimotor strategies: conjugate cross-correlations. AB - A simple method, based on cross-correlation functions (CCFs) between two time series of kinematic or physiological measurements, is proposed for the analysis of multisegmental movements. Special emphasis is placed on measuring accelerations. When the movements of two body segments are coordinated but consistently time lagged, their CCF displays a peak at the corresponding time abscissa. The reproducible positions of the peaks reflect biomechanical or physiological constraints. Several significantly large peaks can be observed in a CCF. It is possible to identify coordinated movements involving more than two segments by applying simple rules of compatibility between the time lags and between the signs of the correlation peaks. With the method proposed, it is possible to determine the signs of relative variation and the time lags of the successive statistically correlated segmental movements. This is particularly useful in the case of both continuous and periodic sensorimotor control, where classical poststimulus methods cannot be applied. Unlike the classical poststimulus methods, this method does not require a time origin, and it is not necessary to monitor the muscles or even to specify exactly which ones are involved. The method is also applicable to experiments involving a time origin (e.g., and applied perturbation), although in this case it is less accurate than the averaging technique. Individual postural strategies can be identified, which suggests some interesting potential applications of the method to clinical studies. PMID- 15753064 TI - The effects of periodic visual occlusion on ball catching. AB - Four experiments were conducted to examine the effects of periodic visual occlusion on one-handed ball catching. Tennis balls were projected one at a time over distances of 8-12 m, and liquid-crystal visual occlusion spectacles provided intermittent vision by opening and closing the lens shutters at different frequencies. As well as frequency, we manipulated duty cycle, or the proportion of time that the lenses were open. Generally, catching performance deteriorated as frequency was reduced. Although longer visual samples (i.e., increased duty cycle) mediated this effect to some extent, the most potent variable was the time between visual samples; performance deteriorated rapidly when this interval was greater than 80 ms (i.e., 10 Hz with 20-ms lens open times). Presumably this occurred because subjects had difficulty integrating visual information separated by longer temporal intervals. PMID- 15753065 TI - Hand preference, practice order, and spatial assimilations in rapid bimanual movement. AB - When subjects make rapid bimanual aiming movements over different distances, spatial assimilations are shown; the shorter distance limb overshoots when paired with a longer distance limb. Recent research has also shown spatial assimilations to be greater in the nonpreferred left limb of right-handed subjects, but it is not known whether the increased spatial assimilations represent a handedness effect or one of hemispheric lateralization of motor control. To determine the nature of the asymmetric effect, left- (n = 32) and right- (n = 60) handed subjects part practiced, then whole practiced, short (20 degrees ) and long 60 degrees ) reversal movements. During whole practice, both groups showed spatial assimilations in the shorter distance limb, particularly when the left limb performed the short movement. This asymmetry was greatest for right-handed subjects, but left-handed subjects showed smaller, but systematic effects, providing moderate support for the hypothesis that the asymmetric effect is due to hemispheric lateralization of motor control. All interlimb differences in spatial accuracy for the short and long movements were eliminated with practice, however, suggesting the asymmetric effect was temporary as well. In addition, subjects who part practiced the long movement just prior to whole practice showed greater overshooting in the short distance limb compared with subjects who followed the other practice order throughout whole practice and the no-KR retention trials. Such findings suggest that the part-practice order of bimanual tasks can directionally bias whole-task performance. PMID- 15753066 TI - Hand, eye, and head coordination while pointing to perturbed targets. AB - Normal human subjects were required to manually point to small visual targets that suddenly changed location upon finger movement initiation. They pointed either as fast or as accurately as possible. Movements of the eyes were measured by electrooculography, and the movements of the unrestrained limb and head were monitored by an optoelectric system (WATSMART), which allowed for the analysis of kinematic parameters in three-dimensional space. The temporal and kinematic reorganization of each body part in response to the target perturbations were variable, which indicated independent control for each part of the system. That is, the timing and nature of the reorganization varied for each body part. In addition, the pattern of reorganization depended upon the speed and accuracy demands of the movement task. As well, the movement termination patterns (eyes finished first, the finger reached the target, then the head stopped moving) were extremely consistent, indicating that movement termination may be a controlled variable. Finally, no evidence was found to suggest that visual information was used to amend arm movements early (before peak velocity) in the trajectory. PMID- 15753067 TI - Crossing without vision of path gaps. AB - Two experiments considered the behavior of subjects who cross gaps in a pathway without benefit of vision. The first experiment contrasted visually and nonvisually guided locomotion, finding a significantly greater number of refusals under nonvisual guidance but no loss of appropriate modulation of step components to the salient dimensions of the actor and layout. Different components of crossing were modified under guidance of different sensory systems; the lengths of the crossing and support spans were primarily altered under nonvisual guidance, and the distance of the support foot to the near edge of the gap was the component of preference under visual guidance. In the second experiment, subjects crossed nonvisually, using probes that varied in length and moment of inertia. The adaptability of the step components to actor and layout dimensions was unchanged, but there was no effect of these mechanical manipulations on the components of crossing identified in Experiment 1. PMID- 15753068 TI - Attention demands in balance recovery following lower limb amputation. AB - The attention demands in balance control after damage to the peripheral sensorimotor system were studied in 12 persons with a recent lower limb amputation. The interference of an arithmetic task with two postural tasks of different complexity (quiet standing and active weight shifting) was examined several times during their rehabilitation while the subjects stood on a force platform. Control data were obtained from healthy subjects. For both postural tasks, persons with amputation performed worse than controls. Quiet standing, a relatively simple task, revealed clear dual-task interference only in the amputation group. Evidence was found for a reduction in dual-task interference as rehabilitation progressed. In contrast, voluntary (feedback-controlled) weight shifting, a more complex task, revealed an equal amount of dual-task interference in persons with amputation and in controls, without changes in interference over the period of rehabilitation. The results indicate that attentional mechanisms may be involved in postural control, depending on both the novelty and complexity of the task. Moreover a reduction in attention demands of quiet standing may reflect a central adaptation of the postural organization to the peripheral sensorimotor impairments caused by lower limb amputation. PMID- 15753069 TI - The stability of precision grip force in older adults. AB - The exceedingly large grip forces that many older adults employ when lifting objects with a precision pinch grip (Cole, 1991) may compensate for a reduced capability to produce a stable isometric force. That is, their grip force may fluctuate enough from moment to moment to yield grip forces that approach the force at which the object would slip from grasp. We examined the within-trial variability of isometric force in old (68-85 years, n = 13) and young (n = 11) human subjects (a) when they were asked to produce a constant pinch force at three target levels (0.49, 2.25, and 10.5 N) with external support of the arm, hand, and force transducer and (b) when they were asked to grasp, lift, and hold a small test object with a precision grip. Pinch force produced in the first task was equally stable across the two subject groups during analysis intervals that lasted 4 s. The elderly subjects produced grip forces when lifting objects that averaged twice as much as those produced by the young subjects. The force variability during the static (hold) phase of the lift for the old subjects was comparable with that used by the young subjects, after adjusting for the difference in grip force. The failure to observe less stable grip force in older adults contradicts a similar recent study. Differences in task (isometric grip force versus isometric abduction torque of a single digit) may account for this conflict, however. Thumb and finger forces for grip are produced through coactivation of many muscles and thus promote smooth force output through temporal summation of twitches. We conclude that peripheral reorganization of muscle in older adults does not yield increased instability of precision grip force and therefore does not contribute directly to increased grip forces in this population. However, force instability may affect other grip configurations (e.g., lateral pinch) or manipulation involving digit abduction or adduction forces. PMID- 15753070 TI - Orienting the finger opposition space during prehension movements. AB - Two experiments are reported that examined the act of prehension when subjects were asked to grasp with their thumb and index finger pads an elongated object resting horizontally on a surface and placed at different orientations with respect to the subject. In Experiment 1, the pad opposition preferences were determined for the six angles of orientation examined. For angles of 90 degrees (object parallel to frontal plane) or less, no rotation of the wrist (pronation) was used; for angles 110 degrees or greater, pronation was systematically employed to reorient the finger opposition space. Only one angle, 100 degrees , produced any evidence of ambiguity in how to grasp the object: Approximately 60% of these grasps involved pronation and 40% did not. Using the foregoing grasp preference data, in Experiment 2 we examined the kinematics of the wrist and elbow trajectories during prehension movements directed at an object in different orientations. Movement time, time to peak acceleration, velocity, and deceleration were measured. No kinematic differences were observed when the object orientation either required (110 degrees ) or did not require (80 degrees ) a pronation. By contrast, if the orientation was changed at the onset of the movement, such that an unpredicted pronation had to be introduced to achieve the grasp, kinematics were affected: Movement time was increased, and the time devoted to deceleration was lengthened. These data are interpreted as evidence that when natural prehension occurs, pronation can be included in the motor plan without affecting the movement kinematics. When constraints are imposed on the movement execution as a consequence of a perturbation, however, the introduction of a pronation component requires kinematic rearrangement. PMID- 15753071 TI - Changes in spinal reflex excitability in a countermanded timed response task. AB - Subjects (N = 8) performed a timed response task in which they attempted to synchronize an impulsive foot-press response with the last in a series of four regularly spaced tones. In Experiment 1, the response was countermanded on one third of the trials (stop trials) by a stop signal that appeared at a predetermined delay after the third tone. No stop signal appeared on the remaining trials (go trials). All subjects showed a systematic transition from withholding the response on stop trials in which the stop signal appeared shortly after the third tone to executing the response on trials in which a single stop signal delay had been chosen so that a response would be made on about 50% of the stop trials. We elicited Hoffmann (H) reflexes from the soleus muscle on all trials to determine whether the reflexes were augmented on occasions on which a response was prepared but withheld. Mean H-reflex amplitudes on go trials and on stop trials on which the response was executed were similar and showed a marked augmentation beginning about 250 ms before response onset; mean H-reflex amplitudes on stop trials on which the response was withheld showed less pronounced augmentation. Inspection of individual H-reflex amplitudes revealed that on stop trials on which the response was withheld the reflexes could be augmented to the same extent as on trials on which the response was executed. This dissociation of H-reflex augmentation and response execution shows that H reflex augmentation reflects a controlled response process. Ballistic response processes therefore must be limited to a brief duration. PMID- 15753072 TI - When can attention not be divided? AB - Humans can guide simple concurrent and seemingly independent movements of digits between hands, but it is shown here that they find it impossible to do this with digits within a hand. This inability demonstrates a basic characteristic of the relation between attention and the control of the skeletal musculature. Mechanisms that are available to provide the illusion of concurrent and independent guidance of separate movements between hands are not available within a single-hand control system. There is no reason to believe that the inability to divide focal attention in the guidance of two concurrent controlled activities, as exemplified in the example of within-hand movements, does not also extend to nonmotor processes. PMID- 15753073 TI - DLPC attenuates alcohol-induced cytotoxicity in HepG2 cells expressing CYP2E1. AB - AIMS: Alcoholic liver injury was shown to result largely from oxidative stress generated by ethanol metabolism via cytochrome P4502E1 (CYP2E1). Our aim was to determine whether this could be overcome by using dilinoleoylphosphatidylcholine (DLPC), an innocuous antioxidant extracted from soybeans. METHODS: To address this question, we determined whether DLPC protects against alcohol-induced cytotoxicity in HepG2 cells expressing CYP2E1. A HepG2 subclone (2E1) expressing CYP2E1 and a control subclone (Neo) were exposed for 2 h to DLPC (10 microM), and then 100 mM ethanol was added for 5 days. RESULTS: Ethanol significantly decreased cell viability in the 2E1 cells and increased apoptosis. These alterations were attenuated by DLPC with the most significant effects in the 2E1 cells. This was accompanied by a reduction of the ethanol-induced oxidative stress, including diminished hydrogen peroxide production in the 2E1, but not in the Neo cells. The mitochondrial membrane potential was significantly diminished by ethanol in both cells. It was also improved after adding DLPC, but only in the 2E1 cells. In these cells, mitochondrial glutathione (GSH) was also partially restored by DLPC, which significantly inhibited the CYP2E1 induction by ethanol. CONCLUSION: DLPC opposes the cytotoxicity induced by alcohol in HepG2 cells expressing CYP2E1, a protective action due, at least in part, to an attenuation of the alcohol-induced oxidative stress and the alteration in the mitochondrial membrane potential. On account of these beneficial effects of DLPC and its innocuity, it is now germane to assess its therapeutic action in alcoholics. PMID- 15753074 TI - A GTP:AMP phosphotransferase, Adk2p, in Saccharomyces cerevisiae. Role of the C terminus in protein folding/stabilization, thermal tolerance, and enzymatic activity. AB - Adenylate kinases participate in maintaining the homeostasis of cellular nucleotides. Depending on the yeast strains, the GTP:AMP phosphotransferase is encoded by the nuclear gene ADK2 with or without a single base pair deletion/insertion near the 3' end of the open reading frame, and the corresponding protein exists as either Adk2p (short) or Adk2p (long) in the mitochondrial matrix. These two forms are identical except that the three C terminal residues of Adk2p (short) are changed in Adk2p (long), and the latter contains an additional nine amino acids at the C terminus of the protein. The short form of Adk2p has so far been considered to be inactive (Schricker, R., Magdolen, V., Strobel, G., Bogengruber, E., Breitenbach, M., and Bandlow, W. (1995) J. Biol. Chem. 270, 31103-31110). Using purified proteins, we show that at the physiological temperature for yeast growth (30 degrees C), both short and long forms of Adk2p are enzymatically active. However, in contrast to the short form, Adk2p (long) is quite resistant to thermal inactivation, urea denaturation, and degradation by trypsin. Unfolding of the long form by high concentrations of urea greatly stimulated its import into isolated mitochondria. Using an integration-based gene-swapping approach, we found that regardless of the yeast strains used, the steady state levels of endogenous Adk2p (long) in mitochondria were 5-10-fold lower compared with those of Adk2p (short). Together, these results suggest that the modified C-terminal domain in Adk2p (long) is not essential for enzyme activity, but it contributes to and strengthens protein folding and/or stability and is particularly important for maintaining enzyme activity under stress conditions. PMID- 15753075 TI - Interaction with type IV pili induces structural changes in the bacterial outer membrane secretin PilQ. AB - Type IV pili are cell surface organelles found on many Gram-negative bacteria. They mediate a variety of functions, including adhesion, twitching motility, and competence for DNA uptake. The type IV pilus is a helical polymer of pilin protein subunits and is capable of rapid polymerization or depolymerization, generating large motor forces in the process. Here we show that a specific interaction between the outer membrane secretin PilQ and the type IV pilus fiber can be detected by far-Western analysis and sucrose density gradient centrifugation. Transmission electron microscopy of preparations of purified pili, to which the purified PilQ oligomer had been added, showed that PilQ was uniquely located at one end of the pilus fiber, effectively forming a "mallet type" structure. Determination of the three-dimensional structure of the PilQ type IV pilus complex at 26-angstroms resolution showed that the cavity within the protein complex was filled. Comparison with a previously determined structure of PilQ at 12-angstroms resolution indicated that binding of the pilus fiber induced a dissociation of the "cap" feature and lateral movement of the "arms" of the PilQ oligomer. The results demonstrate that the PilQ structure exhibits a dynamic response to the binding of its transported substrate and suggest that the secretin could play an active role in type IV pilus assembly as well as secretion. PMID- 15753076 TI - The ATM/p53/p21 pathway influences cell fate decision between apoptosis and senescence in reoxygenated hematopoietic progenitor cells. AB - Hematopoietic cells are often exposed to transient hypoxia as they develop and migrate between blood and tissues. We tested the hypothesis that hypoxia-then reoxygenation represent a stress for hematopoietic progenitor cells. Here we report that reoxygenation-generated oxidative stress induced senescence, tested as staining for SA-beta-galactosidase (SA-beta-gal), of bone marrow progenitor cells. Reoxygenation induced significant DNA damage and inhibited colony formation in lineage-depleted bone marrow cells enriched for progenitor cells. These reoxygenated cells exhibited a prolonged G(0)/G(1) accumulation without significant apoptosis after 24 h of treatments. Reoxygenated bone marrow progenitor cells expressed SA-beta-gal and senescence-associated proteins p53 and p21(WAF1). Reoxygenated Fancc-/- progenitor cells, which underwent significant apoptosis and senescence, tested as staining for SA-beta-gal, also expressed p16(INK4A). Suppression of apoptosis by the pan-caspase inhibitor benzyloxycarbonyl-VAD-fluoromethyl ketone dramatically increased senescent Fancc /- progenitor cells. Senescence induction, tested as staining for SA-beta-gal, in reoxygenated progenitor cells was closely correlated with extent of DNA damage and phosphorylation of ATM at Ser-1981 and p53 at Ser-15. Moreover, inhibition of ATM signaling reduced SA-beta-gal positivity but increased apoptosis of reoxygenated progenitor cells. Thus, these results suggest that the ATM/p53/p21 pathway influences cell fate decision between apoptosis and senescence in reoxygenated hematopoietic progenitor cells. PMID- 15753077 TI - pheA (Rv3838c) of Mycobacterium tuberculosis encodes an allosterically regulated monofunctional prephenate dehydratase that requires both catalytic and regulatory domains for optimum activity. AB - Prephenate dehydratase (PDT) is a key regulatory enzyme in l-phenylalanine biosynthesis. In Mycobacterium tuberculosis, expression of pheA, the gene encoding PDT, has been earlier reported to be iron-dependent (1, 2). We report that M. tuberculosis pheA is also regulated at the protein level by aromatic amino acids. All of the three aromatic amino acids (phenylalanine, tyrosine, and tryptophan) are potent allosteric activators of M. tuberculosis PDT. We also provide in vitro evidence that M. tuberculosis PDT does not possess any chorismate mutase activity, which suggests that, unlike many other enteric bacteria (where PDT exists as a fusion protein with chorismate mutase), M. tuberculosis PDT is a monofunctional and a non-fusion protein. Finally, the biochemical and biophysical properties of the catalytic and regulatory domains (ACT domain) of M. tuberculosis PDT were studied to observe that, in the absence of the ACT domain, the enzyme not only loses its regulatory activity but also its catalytic activity. These novel results provide evidence for a monofunctional prephenate dehydratase enzyme from a pathogenic bacterium that exhibits extensive allosteric activation by aromatic amino acids and is absolutely dependent upon the presence of catalytic as well as the regulatory domains for optimum enzyme activity. PMID- 15753078 TI - Cyclin-dependent kinase inhibitors and JNK act as molecular switches, regulating the choice between growth arrest and apoptosis induced by galectin-8. AB - Galectin-8, a mammalian beta-galactoside binding lectin, functions as an extracellular matrix protein that forms high affinity interactions with integrins. Here we demonstrated that soluble galectin-8 inhibits cell cycle progression and induces growth arrest. These effects cannot be attributed to interference with cell adhesion but can be attributed to a 4-5-fold increase in the cellular content of the cyclin-dependent kinase inhibitor p21, which was already evident following a 4-h incubation of H1299 cells with galectin-8. The increase in p21 levels was preceded by a 3-5-fold increase in JNK and protein kinase B (PKB) activities. Accordingly, SP600125, the inhibitor of JNK, and wortmannin, the inhibitor of phosphatidylinositol 3-kinase, which is the upstream activator of PKB, inhibited the increase in the cellular content of p21. Furthermore, overexpression of a dominant inhibitory form of SEK1, the upstream kinase regulator of JNK, inhibited both JNK activation and p21 accumulation. When p21 expression was inhibited by cycloheximide, galectin-8 directed the cells toward apoptosis, which involves induction of poly(ADP-ribose) polymerase cleavage. Indeed, galectin-8-induced apoptosis was 2-fold higher in HTC (p21 null) cells when compared with parental HTC cells. Because overexpression of galectin-8 attenuates the rate of DNA synthesis, stable colonies that overexpress and secrete galectin-8 can be generated only in cells overexpressing a growth factor receptor, such as the insulin receptor. These results implicate galectin-8 as a modulator of cellular growth through up-regulation of p21. This process involves activation of JNK, which enhances the synthesis of p21, combined with the activation of PKB, which inhibits p21 degradation. These effects of the lectin depended upon protein-sugar interactions and were induced when galectin-8 was present as a soluble ligand or when it was overexpressed in cells. PMID- 15753080 TI - Inhibition of chaperone activity is a shared property of several Cu,Zn-superoxide dismutase mutants that cause amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive motor neuron degeneration, paralysis, and death. Mutant Cu,Zn superoxide dismutase (SOD1) causes a subset of ALS by an unidentified toxic property. Increasing evidence suggests that chaperone dysfunction plays a role in motor neuron degeneration in ALS. To investigate the relationship between mutant SOD1 expression and chaperone dysfunction, we measured chaperone function in central nervous system tissue lysates from normal mice and transgenic mice expressing human SOD1 variants. We observed a significant decrease in chaperone activity in tissues from mice expressing ALS-linked mutant SOD1 but not control mice expressing human wild type SOD1. This decrease was detected only in the spinal cord, became apparent by 60 days of age (before the onset of muscle weakness and significant motor neuron loss), and persisted throughout the late stages. In addition, this impairment of chaperone activity occurred only in cytosolic but not in mitochondrial and nuclear fractions. Furthermore, multiple recombinant human SOD1 mutants with differing biochemical and biophysical properties inhibited chaperone function in a cell-free extract of normal mouse spinal cords. Thus, mutant SOD1 proteins may impair chaperone function independent of gene expression in vivo, and this inhibition may be a shared property of ALS-linked mutant SOD1 proteins. PMID- 15753079 TI - Epithelial sodium channel inhibition by AMP-activated protein kinase in oocytes and polarized renal epithelial cells. AB - The epithelial Na(+) channel (ENaC) regulates epithelial salt and water reabsorption, processes that require significant expenditure of cellular energy. To test whether the ubiquitous metabolic sensor AMP-activated kinase (AMPK) regulates ENaC, we examined the effects of AMPK activation on amiloride-sensitive currents in Xenopus oocytes and polarized mouse collecting duct mpkCCD(c14) cells. Microinjection of oocytes expressing mouse ENaC (mENaC) with either active AMPK protein or an AMPK activator inhibited mENaC currents relative to controls as measured by two-electrode voltage-clamp studies. Similarly, pharmacological AMPK activation or overexpression of an activating AMPK mutant in mpkCCD(c14) cells inhibited amiloride-sensitive short circuit currents. Expression of a degenerin mutant beta-mENaC subunit (S518K) along with wild type alpha and gamma increased the channel open probability (P(o)) to approximately 1. However, AMPK activation inhibited currents similarly with expression of either degenerin mutant or wild type mENaC. Single channel recordings under these conditions demonstrated that neither P(o) nor channel conductance was affected by AMPK activation. Moreover, expression of a Liddle's syndrome-type beta-mENaC mutant (Y618A) greatly enhanced ENaC whole cell currents relative to wild type ENaC controls and prevented AMPK-dependent inhibition. These findings indicate that AMPK-dependent ENaC inhibition is mediated through a decrease in the number of active channels at the plasma membrane (N), presumably through enhanced Nedd4-2 dependent ENaC endocytosis. The AMPK-ENaC interaction appears to be indirect; AMPK did not bind ENaC in cells, as assessed by in vivo pull-down assays, nor did it phosphorylate ENaC in vitro. In summary, these results suggest a novel mechanism for coupling ENaC activity and renal Na(+) handling to cellular metabolic status through AMPK, which may help prevent cellular Na(+) loading under hypoxic or ischemic conditions. PMID- 15753081 TI - Prediction of collagen stability from amino acid sequence. AB - An algorithm was derived to relate the amino acid sequence of a collagen triple helix to its thermal stability. This calculation is based on the triple helical stabilization propensities of individual residues and their intermolecular and intramolecular interactions, as quantitated by melting temperature values of host guest peptides. Experimental melting temperature values of a number of triple helical peptides of varying length and sequence were successfully predicted by this algorithm. However, predicted T(m) values are significantly higher than experimental values when there are strings of oppositely charged residues or concentrations of like charges near the terminus. Application of the algorithm to collagen sequences highlights regions of unusually high or low stability, and these regions often correlate with biologically significant features. The prediction of stability from sequence indicates an understanding of the major forces maintaining this protein motif. The use of highly favorable KGE and KGD sequences is seen to complement the stabilizing effects of imino acids in modulating stability and may become dominant in the collagenous domains of bacterial proteins that lack hydroxyproline. The effect of single amino acid mutations in the X and Y positions can be evaluated with this algorithm. An interactive collagen stability calculator based on this algorithm is available online. PMID- 15753083 TI - 1.6-Angstroms crystal structure of EntA-im. A bacterial immunity protein conferring immunity to the antimicrobial activity of the pediocin-like bacteriocin enterocin A. AB - Many Gram-positive bacteria produce ribosomally synthesized antimicrobial peptides, often termed bacteriocins. Genes encoding pediocin-like bacteriocins are generally cotranscribed with or in close vicinity to a gene encoding a cognate immunity protein that protects the bacteriocin-producer from their own bacteriocin. We present the first crystal structure of a pediocin-like immunity protein, EntA-im, conferring immunity to the bacteriocin enterocin A. Determination of the structure of this 103-amino acid protein revealed that it folds into an antiparallel four-helix bundle with a flexible C-terminal part. The fact that the immunity protein conferring immunity to carnobacteriocin B2 also consists of a four-helix bundle (Sprules, T., Kawulka, K. E., and Vederas, J. C. (2004) Biochemistry 43, 11740-11749) strongly indicates that this is a conserved structural motif in all pediocin-like immunity proteins. The C-terminal half of the immunity protein contains a region that recognizes the C-terminal half of the cognate bacteriocin, and the flexibility in the C-terminal end of the immunity protein might thus be an important characteristic that enables the immunity protein to interact with its cognate bacteriocin. By homology modeling of three other pediocin-like immunity proteins and calculation of the surface charge distribution for EntA-im and the three structure models, different charge distributions were observed. The differences in the latter part of helix 3, the beginning of helix 4, and the loop connecting these helices might also be of importance in determining the specificity. PMID- 15753082 TI - The benzo[c]phenanthridine alkaloid, sanguinarine, is a selective, cell-active inhibitor of mitogen-activated protein kinase phosphatase-1. AB - Mitogen-activated protein kinase phosphatase-1 (MKP-1) is a dual specificity phosphatase that is overexpressed in many human tumors and can protect cells from apoptosis caused by DNA-damaging agents or cellular stress. Small molecule inhibitors of MKP-1 have not been reported, in part because of the lack of structural guidance for inhibitor design and definitive assays for MKP-1 inhibition in intact cells. Herein we have exploited a high content chemical complementation assay to analyze a diverse collection of pure natural products for cellular MKP-1 inhibition. Using two-dimensional Kolmogorov-Smirnov statistics, we identified sanguinarine, a plant alkaloid with known antibiotic and antitumor activity but no primary cellular target, as a potent and selective inhibitor of MKP-1. Sanguinarine inhibited cellular MKP-1 with an IC50 of 10 microM and showed selectivity for MKP-1 over MKP-3. Sanguinarine also inhibited MKP-1 and the MKP-1 like phosphatase, MKP-L, in vitro with IC50 values of 17.3 and 12.5 microM, respectively, and showed 5-10-fold selectivity for MKP-3 and MKP 1 over VH-1-related phosphatase, Cdc25B2, or protein-tyrosine phosphatase 1B. In a human tumor cell line with high MKP-1 levels, sanguinarine caused enhanced ERK and JNK/SAPK phosphorylation. A close congener of sanguinarine, chelerythrine, also inhibited MKP-1 in vitro and in whole cells, and activated ERK and JNK/SAPK. In contrast, sanguinarine analogs lacking the benzophenanthridine scaffold did not inhibit MKP-1 in vitro or in cells nor did they cause ERK or JNK/SAPK phosphorylation. These data illustrate the utility of a chemical complementation assay linked with multiparameter high content cellular screening. PMID- 15753084 TI - The role of mitochondrially bound arginase in the regulation of urea synthesis: studies with [U-15N4]arginine, isolated mitochondria, and perfused rat liver. AB - The main goal of the current study was to elucidate the role of mitochondrial arginine metabolism in the regulation of N-acetylglutamate and urea synthesis. We hypothesized that arginine catabolism via mitochondrially bound arginase augments ureagenesis by supplying ornithine for net synthesis of citrulline, glutamate, N acetylglutamate, and aspartate. [U-(15)N(4)]arginine was used as precursor and isolated mitochondria or liver perfusion as a model system to monitor arginine catabolism and the incorporation of (15)N into various intermediate metabolites of the urea cycle. The results indicate that approximately 8% of total mitochondrial arginase activity is located in the matrix, and 90% is located in the outer membrane. Experiments with isolated mitochondria showed that approximately 60-70% of external [U-(15)N(4)]arginine catabolism was recovered as (15)N-labeled ornithine, glutamate, N-acetylglutamate, citrulline, and aspartate. The production of (15)N-labeled metabolites was time- and dose-dependent. During liver perfusion, urea containing one (U(m+1)) or two (U(m+2)) (15)N was generated from perfusate [U-(15)N(4)]arginine. The output of U(m+2) was between 3 and 8% of total urea, consistent with the percentage of activity of matrix arginase. U(m+1) was formed following mitochondrial production of [(15)N]glutamate from [alpha,delta-(15)N(2)]ornithine and transamination of [(15)N]glutamate to [(15)N]aspartate. The latter is transported to cytosol and incorporated into argininosuccinate. Approximately 70, 75, 7, and 5% of hepatic ornithine, citrulline, N-acetylglutamate, and aspartate, respectively, were derived from perfusate [U-(15)N(4)]arginine. The results substantiate the hypothesis that intramitochondrial arginase, presumably the arginase-II isozyme, may play an important role in the regulation of hepatic ureagenesis by furnishing ornithine for net synthesis of N-acetylglutamate, citrulline, and aspartate. PMID- 15753086 TI - Human tumorous imaginal disc 1 (TID1) associates with Trk receptor tyrosine kinases and regulates neurite outgrowth in nnr5-TrkA cells. AB - The human tumorous imaginal disc 1 (TID1) proteins including TID1(L) and TID1(S), members of the DnaJ domain protein family, are involved in multiple intracellular signaling pathways such as apoptosis induction, cell proliferation, and survival. Here we report that TID1 associates with the Trk receptor tyrosine kinases and regulates nerve growth factor (NGF)-induced neurite outgrowth in PC12-derived nnr5 cells. Binding assays and transfection studies showed that the carboxyl terminal end of TID1 (residues 224-429) bound to Trk at the activation loop (Tyr(P)(683)-Tyr(684)(P)(684) in rat TrkA) and that TID1 was tyrosine phosphorylated by Trk both in yeast and in transfected cells. Moreover endogenous TID1 was also tyrosine phosphorylated by and co-immunoprecipitated with Trk in neurotrophin-stimulated primary rat hippocampal neurons. Overexpression studies showed that both TID1(L) and TID1(S) significantly facilitated NGF-induced neurite outgrowth in TrkA-expressing nnr5 cells possibly through a mechanism involving increased activation of mitogen-activated protein kinase. Consistently knockdown of endogenous TID1, mediated with specific short hairpin RNA, significantly reduced NGF-induced neurite growth in nnr5-TrkA cells. These data provide the first evidence that TID1 is a novel intracellular adaptor that interacts with the Trk receptor tyrosine kinases in an activity-dependent manner to facilitate Trk-dependent intracellular signaling. PMID- 15753085 TI - A novel protein kinase B (PKB)/AKT-binding protein enhances PKB kinase activity and regulates DNA synthesis. AB - Protein kinase B (PKB)/Akt reportedly plays a role in the survival and/or proliferation of cells. We identified a novel protein, which binds to PKB, using a yeast two-hybrid screening system. This association was demonstrated not only in vivo by overexpressing both proteins or by coimmunoprecipitation of the endogenous proteins, but also in vitro using glutathione S-transferase fusion proteins. Importantly, this protein specifically associates with the C terminus of PKB but not with other AGC kinases and enhances PKB phosphorylation and kinase activation without growth factor stimulation. Thus, we termed this Akt-specific binding protein APE (Akt-phosphorylation enhancer). Since APE-induced phosphorylation of PKB did not occur in cells treated with wortmannin or LY294002, APE itself is not a kinase but seems to enhance or prolong the phosphoinositide 3-kinase-dependent phosphorylation of PKB. In cells in which APE was suppressed by small interfering RNA, DNA synthesis was significantly reduced with suppression of PKB phosphorylation, suggesting a synergistic role of APE in PKB-induced proliferation. On the other hand, in cells overexpressing both PKB and APE, despite markedly increased basal phosphorylation of PKB, both DNA rereplication and subsequent Chk2 phosphorylation and apoptosis were seen, suggesting the involvement of APE in the regulation of cell cycling replication licensing. Taking these observations together, APE appears to be a novel regulator of PKB phosphorylation. Furthermore, the interaction between APE and PKB, possibly dependent on the expression levels of both proteins, may be a novel molecular mechanism leading to proliferation and/or apoptosis. PMID- 15753087 TI - From ATP as substrate to ADP as coenzyme: functional evolution of the nucleotide binding subunit of dihydroxyacetone kinases. AB - Dihydroxyacetone kinases are a family of sequence-related enzymes that utilize either ATP or a protein of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) as a source of high energy phosphate. The PTS is a multicomponent system involved in carbohydrate uptake and control of carbon metabolism in bacteria. Phylogenetic analysis suggests that the PTS-dependent dihydroxyacetone kinases evolved from an ATP-dependent ancestor. Their nucleotide binding subunit, an eight-helix barrel of regular up-down topology, retains ADP as phosphorylation site for the double displacement of phosphate from a phospho-histidine of the PTS protein to dihydroxyacetone. ADP is bound essentially irreversibly with a t((1/2)) of 100 min. Complexation with ADP increases the thermal unfolding temperature of dihydroxyacetone L from 40 (apo-form) to 65 degrees C (holoenzyme). ADP assumes the same role as histidines, cysteines, and aspartic acids in histidine kinases and PTS proteins. This conversion of a substrate binding site into a cofactor binding site reflects a remarkable instance of parsimonious evolution. PMID- 15753088 TI - The RNA-binding protein IMP-3 is a translational activator of insulin-like growth factor II leader-3 mRNA during proliferation of human K562 leukemia cells. AB - IMP-3, a member of the insulin-like growth factor-II (IGF-II) mRNA-binding protein (IMP) family, is expressed mainly during embryonic development and in some tumors. Thus, IMP-3 is considered to be an oncofetal protein. The functional significance of IMP-3 is not clear. To identify the functions of IMP-3 in target gene expression and cell proliferation, RNA interference was employed to knock down IMP-3 expression. Using human K562 leukemia cells as a model, we show that IMP-3 protein associates with IGF-II leader-3 and leader-4 mRNAs and H19 RNA but not c-myc and beta-actin mRNAs in vivo by messenger ribonucleoprotein immunoprecipitation analyses. IMP-3 knock down significantly decreased levels of intracellular and secreted IGF-II without affecting IGF-II leader-3, leader-4, c myc, or beta-actin mRNA levels and H19 RNA levels compared with the negative control siRNA treatment. Moreover, IMP-3 knock down specifically suppressed translation of chimeric IGF-II leader-3/luciferase mRNA without altering reporter mRNA levels. Together, these results suggest that IMP-3 knock down reduced IGF-II expression by inhibiting translation of IGF-II mRNA. IMP-3 knock down also markedly inhibited cell proliferation. The addition of recombinant human IGF-II peptide to these cells restored cell proliferation rates to normal. IMP-3 and IMP 1, two members of the IMP family with significant structural similarity, appear to have some distinct RNA targets and functions in K562 cells. Thus, we have identified IMP-3 as a translational activator of IGF-II leader-3 mRNA. IMP-3 plays a critical role in regulation of cell proliferation via an IGF-II-dependent pathway in K562 leukemia cells. PMID- 15753089 TI - Palmitoylation and intracellular domain interactions both contribute to raft targeting of linker for activation of T cells. AB - Some transmembrane proteins must associate with lipid rafts to function. However, even if acylated, transmembrane proteins should not pack well with ordered raft lipids, and raft targeting is puzzling. Acylation is necessary for raft targeting of linker for activation of T cells (LAT). To determine whether an acylated transmembrane domain is sufficient, we examined raft association of palmitoylated and nonpalmitoylated LAT transmembrane peptides in lipid vesicles by a fluorescence quenching assay, by microscopic examination, and by association with detergent-resistant membranes (DRMs). All three assays detected very low raft association of the nonacylated LAT peptide. DRM association was the same as a control random transmembrane peptide. Acylation did not measurably enhance raft association by the first two assays but slightly enhanced DRM association. The palmitoylated LAT peptide and a FLAG-tagged LAT transmembrane domain construct expressed in cells showed similar DRM association when both were reconstituted into mixed vesicles (containing cell-derived proteins and lipids and excess artificial raft-forming lipids) before detergent extraction. We conclude that the acylated LAT transmembrane domain has low inherent raft affinity. Full-length LAT in mixed vesicles associated better with DRMs than the peptide. However, cells appeared to contain two pools of LAT, with very different raft affinities. Since some LAT (but not the transmembrane domain construct) was isolated in a protein complex, and the Myc- and FLAG-tagged forms of LAT could be mutually co immunoprecipitated, oligomerization or interactions with other proteins may enhance raft affinity of one pool of LAT. We conclude that both acylation and other factors, possibly protein-protein interactions, target LAT to rafts. PMID- 15753090 TI - Leader peptide-mediated transcriptional attenuation of lysine biosynthetic gene cluster in Thermus thermophilus. AB - The molecular mechanism for regulation of the genes involved in the biosynthesis of amino acids is poorly identified in Thermus thermophilus. In this study, we analyzed the transcriptional control of the major lysine biosynthetic gene cluster in T. thermophilus. S1 nuclease mapping revealed that the transcription, which is repressed by lysine, starts at 111 bp, upstream of the translational start codon, ATG, for the homocitrate synthase (hcs) gene. The 5'-leader region of 111 bp carries a sequence that can encode a short peptide of 14 amino acids with tandem-arranged lysine residues in its sequence. The nucleotide sequence of the region suggests that the transcript can form complicated secondary structures. Deletion of most of the 5'-leader region or mutation of the tandem lysine codons suppressed the transcriptional repression by lysine. Mutation of the tandem codons from lysine to glutamine resulted in glutamine-dependent repression of the gene connected downstream, indicating that the leader peptide mediated the transcriptional attenuation of the gene expression. This is the first report demonstrating the transcriptional regulation of amino acid biosynthesis in T. thermophilus. PMID- 15753091 TI - GRIM-19 interacts with nucleotide oligomerization domain 2 and serves as downstream effector of anti-bacterial function in intestinal epithelial cells. AB - Nucleotide oligomerization domain 2 (NOD2) functions as a mammalian cytosolic pathogen recognition molecule, and variants have been associated with risk for Crohn disease. We recently demonstrated that NOD2 functions as an anti-bacterial factor limiting survival of intracellular invasive bacteria. To gain further insight into the mechanism of NOD2 activation and signal transduction, we performed yeast two-hybrid screening. We demonstrate that GRIM-19, a protein with homology to the NADPH dehydrogenase complex, interacts with endogenous NOD2 in HT29 cells. GRIM-19 is required for NF-kappaB activation following NOD2-mediated recognition of bacterial muramyl dipeptide. GRIM-19 also controls pathogen invasion of intestinal epithelial cells. GRIM-19 expression is decreased in inflamed mucosa of patients with inflammatory bowel diseases. GRIM-19 may be a key component in NOD2-mediated innate mucosal responses and serve to regulate intestinal epithelial cell responses to microbes. PMID- 15753092 TI - Novel role for amphiregulin in protection from liver injury. AB - Clinically, the Fas and Fas ligand system plays a central role in the development of hepatocyte apoptosis, a process contributing to a broad spectrum of liver diseases. Therefore, the development of therapies aimed at the inhibition of hepatocyte apoptosis is a major issue. Activation of the epidermal growth factor receptor has been shown to convey survival signals to the hepatocyte. To learn about the endogenous response of epidermal growth factor receptor ligands during Fas-mediated liver injury we investigated the expression of epidermal growth factor, transforming growth factor alpha, heparin-binding epidermal growth factor like growth factor, betacellulin, epiregulin, and amphiregulin in the liver of mice challenged with Fas-agonist antibody. Amphiregulin expression, barely detectable in healthy liver, was significantly up-regulated. Amphiregulin administration abrogated Fas-mediated liver injury in mice and showed direct anti apoptotic effects in primary hepatocytes. Amphiregulin activated the Akt and signal transducer and activator of transcription-3 survival pathways, and up regulated Bcl-xL expression. Amphiregulin knock-out mice showed signs of chronic liver damage in the absence of any noxious treatment, and died faster than wild type mice in response to lethal doses of Fas-agonist antibody. In contrast, these mice were more resistant against sublethal liver damage, supporting the hypothesis that chronic liver injury can precondition hepatocytes inducing resistance to subsequent cell death. These results show that amphiregulin is a protective factor induced in response to liver damage and that it may be therapeutic in liver diseases. PMID- 15753093 TI - N-myristoyltransferase 1 is essential in early mouse development. AB - N-Myristoyltransferase (NMT) transfers myristate to an amino-terminal glycine of many eukaryotic proteins. In yeast, worms, and flies, this enzyme is essential for viability of the organism. Humans and mice possess two distinct but structurally similar enzymes, NMT1 and NMT2. These two enzymes have similar peptide specificities, but no one has examined the functional importance of the enzymes in vivo. To address this issue, we performed both genetic and biochemical studies. Northern blots with RNA from adult mice and in situ hybridization studies of day 13.5 embryos revealed widespread expression of both Nmt1 and Nmt2. To determine whether the two enzymes are functionally redundant, we generated Nmt1-deficient mice carrying a beta-galactosidase marker gene. beta-Galactosidase staining of tissues from heterozygous Nmt1-deficient (Nmt1+/-) mice and embryos confirmed widespread expression of Nmt1. Intercrosses of Nmt1+/- mice yielded no viable homozygotes (Nmt1-/-), and heterozygotes were born at a less than predicted frequency. Nmt1-/- embryos died between embryonic days 3.5 and 7.5. Northern blots revealed lower levels of Nmt2 expression in early development than at later time points, a potential explanation for the demise of Nmt1-/- embryos. To explore this concept, we generated Nmt1-/- embryonic stem (ES) cells. The Nmt2 mRNA could be detected in Nmt1-/- ES cells, but the total NMT activity levels were reduced by approximately 95%, suggesting that Nmt2 contributes little to total enzyme activity levels in these early embryo cells. The Nmt1-/- ES cells were functionally abnormal; they yielded small embryoid bodies in in vitro differentiation experiments and did not contribute normally to organogenesis in chimeric mice. We conclude that Nmt1 is not essential for the viability of mammalian cells but is required for development, likely because it is the principal N-myristoyltransferase in early embryogenesis. PMID- 15753094 TI - An insulin-response element-binding protein that ameliorates hyperglycemia in diabetes. AB - Insulin modulates glucose homeostasis, but the role of insulin-responsive transcription factors in such actions is not well understood. Recently, we have identified the insulin-response element-binding protein-1 (IRE-BP1) as a transcription factor that appears to mediate insulin action on multiple target genes. To examine the possibility that IRE-BP1 is an insulin-responsive glucoregulatory factor involved in the metabolic actions of insulin, we investigated the effect of adenoviral overexpression of hepatic IRE-BP1 on the glycemic control of insulin-resistant diabetic rats. Adenoviral IRE-BP1 lowered both fasting and postprandial glucose levels, and microarray of hepatic RNA revealed modulation of the expression of genes involved in gluconeogenesis, lipogenesis, and fatty acid oxidation. The insulin mimetic effects of IRE-BP1 were also confirmed in L6 myocytes; stable constitutive expressions of IRE-BP1 enhanced glucose transporter expression, glucose uptake, and glycogen accumulation in these cells. These findings showed physiologic sufficiency of IRE BP1 as the transcriptional mediator of the metabolic action of insulin. Understanding IRE-BP1 action should constitute a useful probe into the mechanisms of metabolic regulation and an important target to develop therapeutic agents that mimic or enhance insulin action. PMID- 15753095 TI - Transcriptional blockade induces p53-dependent apoptosis associated with translocation of p53 to mitochondria. AB - The tumor suppressor p53 functions as a transcriptional activator to induce cell cycle arrest and apoptosis in response to DNA damage. Although p53 was also shown to mediate apoptosis in a manner independent of its transactivation activity, the mechanism and conditions that trigger such cell death have remained largely unknown. We have now shown that inhibition of RNA polymerase II-mediated transcription by alpha-amanitin or RNA interference induced p53-dependent apoptosis. Inhibition of pol II-mediated transcription resulted in down regulation of p21Cip1, which was caused by both transcriptional suppression and protein degradation, despite eliciting p53 accumulation, allowing the cells to progress into S phase and then to undergo apoptosis. This cell death did not require the transcription of p53 target genes and was preceded by translocation of the accumulated p53 to mitochondria. Our data thus suggested that blockade of pol II-mediated transcription induced p53 accumulation in mitochondria and was the critical factor for eliciting p53-dependent but transcription-independent apoptosis. PMID- 15753096 TI - Low density lipoprotein receptor-related protein 1 (LRP1) controls endocytosis and c-CBL-mediated ubiquitination of the platelet-derived growth factor receptor beta (PDGFR beta). AB - The low density lipoprotein receptor-related protein 1 (LRP1) has been implicated in intracellular signaling functions as well as in lipid metabolism. Recent in vivo and in vitro studies suggest that LRP1 is a physiological modulator of the platelet-derived growth factor (PDGF) signaling pathway. Here we show that in mouse fibroblasts LRP1 modulates PDGF-BB signaling by controlling endocytosis and ligand-induced down-regulation of the PDGF receptor beta (PDGFRbeta). In LRP1 deficient fibroblasts, basal PDGFRbeta tyrosine kinase activity was derepressed, and PDGF-BB-induced endocytosis and degradation of PDGFRbeta were accelerated as compared with control cells. This was accompanied by rapid uptake of receptor bound PDGF-BB into the cells and by attenuated ERK activation in response to PDGF BB stimulation. Pulse-chase analysis indicated that the steady-state turnover rate of PDGFRbeta was also accelerated in LRP-deficient fibroblasts. The rapid degradation of PDGFRbeta in the LRP1-deficient fibroblasts was prevented by MG132 and chloroquine. Furthermore, the association of PDGFRbeta with c-Cbl, a ubiquitin E3-ligase, as well as the ligand-induced ubiquitination of PDGFRbeta were increased in LRP1-deficient fibroblasts. We show that LRP1 can directly interact with c-Cbl, suggesting a Sprouty-like role for LRP1 in regulating the access of the PDGFRbeta to the ubiquitination machinery. Thus, LRP1 modulates PDGF signaling by controlling ubiquitination and endocytosis of the PDGFRbeta. PMID- 15753097 TI - Mammalian prion protein suppresses Bax-induced cell death in yeast. AB - Several lines of evidence suggest that PrP(C), the non-infectious form of the prion protein, may function to protect neurons and other cells from stress or toxicity. In this paper, we report on the use of the yeast Saccharomyces cerevisiae as a model system to assay the cytoprotective activity of PrP(C). The mammalian pro-apoptotic protein, Bax, confers a lethal phenotype when expressed in yeast. Since overexpression of PrP(C) has been found to prevent Bax-mediated cell death in cultured human neurons, we explored whether PrP could also suppress Bax-induced cell death in yeast. We utilized a form of mouse PrP containing a modified signal peptide that we had previously shown is efficiently targeted to the secretory pathway in yeast. We found that this PrP potently suppressed the death of yeast cells expressing mammalian Bax under control of a galactose inducible promoter. In contrast, cytosolic PrP-(23-231) failed to rescue growth of Bax-expressing yeast, indicating that protective activity requires targeting of PrP to the secretory pathway. Deletion of the octapeptide repeat region did not affect the rescuing activity of PrP, but deletion of a charged region encompassing residues 23-31 partially eliminated activity. We also tested several PrP mutants associated with human familial prion diseases and found that only a mutant containing nine extra octapeptide repeats failed to suppress Bax-induced cell death. These findings establish a simple and genetically tractable system for assaying a putative biological activity of PrP(C). PMID- 15753098 TI - Crystal structure of a nicotinate phosphoribosyltransferase from Thermoplasma acidophilum. AB - We have determined the crystal structure of nicotinate phosphoribosyltransferase from Themoplasma acidophilum (TaNAPRTase). The TaNAPRTase has three domains, an N terminal domain, a central functional domain, and a unique C-terminal domain. The crystal structure revealed that the functional domain has a type II phosphoribosyltransferase fold that may be a common architecture for both nicotinic acid and quinolinic acid (QA) phosphoribosyltransferases (PRTase) despite low sequence similarity between them. Unlike QAPRTase, TaNAPRTase has a unique extra C-terminal domain containing a zinc knuckle-like motif containing 4 cysteines. The TaNAPRTase forms a trimer of dimers in the crystal. The active site pocket is formed at dimer interfaces. The complex structures with phosphoribosylpyrophosphate (PRPP) and nicotinate mononucleotide (NAMN) showed, surprisingly, that functional residues lining on the active site of TaNAPRTase are quite different from those of QAPRTase, although their substrates are quite similar to each other. The phosphate moiety of PRPP and NAMN is anchored to the phosphate-binding loops formed by backbone amides, as found in many alpha/beta barrel enzymes. The pyrophosphate moiety of PRPP is located at the entrance of the active site pocket, whereas the nicotinate moiety of NAMN is located deep inside. Interestingly, the nicotinate moiety of NAMN is intercalated between highly conserved aromatic residues Tyr(21) and Phe(138). Careful structural analyses combined with other NAPRTase sequence subfamilies reveal that TaNAPRTase represents a unique sequence subfamily of NAPRTase. The structures of TaNAPRTase also provide valuable insight for other sequence subfamilies such as pre-B cell colony-enhancing factor, known to have nicotinamide phosphoribosyltransferase activity. PMID- 15753099 TI - Structure and function of carbonic anhydrases from Mycobacterium tuberculosis. AB - Carbonic anhydrases catalyze the reversible hydration of carbon dioxide to form bicarbonate. This activity is universally required for fatty acid biosynthesis as well as for the production of a number of small molecules, pH homeostasis, and other functions. At least three different carbonic anhydrase families are known to exist, of which the alpha-class found in humans has been studied in most detail. In the present work, we describe the structures of two of the three beta class carbonic anhydrases that have been identified in Mycobacterium tuberculosis, i.e. Rv1284 and Rv3588c. Both structures were solved by molecular replacement and then refined to resolutions of 2.0 and 1.75 A, respectively. The active site of Rv1284 is small and almost completely shielded from solvent, whereas that of Rv3588c is larger and quite open to solution. Differences in coordination of the active site metal are also observed. In Rv3588c, an aspartic acid side chain displaces a water molecule and coordinates directly to the zinc ion, thereby closing the zinc coordination sphere and breaking the salt link to a nearby arginine that is a feature of Rv1284. The two carbonic anhydrases thus exhibit both of the metal coordination geometries that have previously been observed for structures in this family. Activity studies demonstrate that Rv3588c is a completely functional carbonic anhydrase. The apparent lack of activity of Rv1284 in the present assay system is likely exacerbated by the observed depletion of zinc in the preparation. PMID- 15753100 TI - Crystal structure of the N-terminal domain of the group B streptococcus alpha C protein. AB - Group B Streptococcus (GBS) is the leading cause of bacterial pneumonia, sepsis, and meningitis among neonates and an important cause of morbidity among pregnant women and immunocompromised adults. Invasive diseases due to GBS are attributed to the ability of the pathogen to translocate across human epithelial surfaces. The alpha C protein (ACP) has been identified as an invasin that plays a role in internalization and translocation of GBS across epithelial cells. The soluble N terminal domain of ACP (NtACP) blocks the internalization of GBS. We determined the 1.86-A resolution crystal structure of NtACP comprising residues Ser(52) through Leu(225) of the full-length ACP. NtACP has two domains, an N-terminal beta-sandwich and a C-terminal three-helix bundle. Structural and topological alignments reveal that the beta-sandwich shares structural elements with the type III fibronectin fold (FnIII), but includes structural elaborations that make it unique. We have identified a potential integrin-binding motif consisting of Lys Thr-Asp(146), Arg(110), and Asp(118). A similar arrangement of charged residues has been described in other invasins. ACP shows a heparin binding activity that requires NtACP. We propose a possible heparin-binding site, including one surface of the three-helix bundle, and nearby portions of the sandwich and repeat domains. We have validated this prediction using assays of the heparin binding and cell-adhesion properties of engineered fragments of ACP. This is the first crystal structure of a member of the highly conserved Gram-positive surface alpha like protein family, and it will enable the internalization mechanism of GBS to be dissected at the atomic level. PMID- 15753101 TI - A putative function for the arabidopsis Fe-Phytosiderophore transporter homolog AtYSL2 in Fe and Zn homeostasis. AB - Although Arabidopsis thaliana does not produce phytosiderophores (PS) under Fe deficiency, it contains eight homologs of the metal-PS/metal-nicotianamine (NA) transporter ZmYS1 from maize. This study aimed to investigate whether one of the closest Arabidopsis homologs to ZmYS1, AtYSL2, is involved in metal-chelate transport. Northern analysis revealed high expression levels of AtYSL2 in Fe sufficient or Fe-resupplied roots, while under Fe deficiency transcript levels decreased. Quantitative real-time polymerase chain reaction (PCR) and analysis of transgenic plants expressing an AtYSL2 promoter::beta-glucuronidase gene further allowed the detection of down-regulated AtYSL2 gene expression under Zn and Fe deficiency. In contrast to ZmYS1, AtYSL2 did not mediate metal-PS or metal-NA transport in yeast mutants defective in Cu or Fe uptake, nor did AtYSL2 mediate Fe(II)-NA-, Fe(III)-NA- or Ni(II)-NA-inducible currents when assayed by two electrode voltage clamp in Xenopus oocytes. Moreover, truncation of the N terminus to remove putative phosphorylation sites that might trigger autoinhibition did not confer functionality to AtYSL2. A direct growth comparison of yeast cells transformed with AtYSL2 in two different yeast expression vectors showed that transformation with empty pFL61 repressed growth even under non limiting Fe supply. We therefore conclude that the yeast complementation assay previously employed does not allow the identification of AtYSL2 as an Fe-NA transporter. Transgenic plants expressing an AtYSL2 promoter::beta-glucuronidase gene showed expression in root endodermis and pericycle cells facing the meta xylem tubes. Taken together, our investigations support an involvement of AtYSL2 in Fe and Zn homeostasis, although functionality or substrate specificity are likely to differ between AtYSL2 and ZmYS1. PMID- 15753102 TI - The formation of Zn-Chl a in Chlorella heterotrophically grown in the dark with an excessive amount of Zn2+. AB - Chlorella, when heterotrophically cultivated in the dark, is able to grow with Zn2+ at 10-40 mM, which is 10 times the concentration lethal to autotrophically grown cells. However, the lag phase is prolonged with increasing concentrations of Zn2+; for example, in this study, 1 d of the control lag phase was prolonged to about 16 d with Zn2+ at 16.7 mM (x2,000 of the control). Once the cells started to grow, the log phase was finished within 4-6 d regardless of Zn concentration, which was almost the same as that of the control. The photosysystem I reaction center chlorophyll, P700, and the far-red fluorescence were detected only after the late log phase of the growth curve, suggesting that chlorophyll-protein complexes can be organized after cell division has ceased. Interestingly, at more than 16.7 mM of Zn2+, Zn-chlorophyll a was accumulated and finally accounted for about 25% of the total chlorophyll a in the late stationary phase. We found that the Zn-chlorophyll a was present in the thylakoid membranes and not in the soluble fractions of the cells. The rather low fluorescence yield at around 680 nm in the stationary phase suggests that Zn-chlorophyll a can transfer its excitation energy to other chlorophylls. Before accumulation of Zn chlorophyll a, a marked amount of pheophytin a was temporally accumulated, suggesting that Zn-chlorophyll a could be chemically synthesized via pheophytin a. PMID- 15753104 TI - The promoter for C4-type mitochondrial aspartate aminotransferase does not direct bundle sheath-specific expression in transgenic rice plants. AB - For NAD-malic enzyme (NAD-ME)-type C4 photosynthesis, two types of aspartate aminotransferase (AAT) are involved. We examined the expression pattern of the Panicum miliaceum mitochondrial Aat gene (PmAat) and P. miliaceum cytosolic Aat gene (PcAat) in transgenic rice plants, which were specifically expressed in bundle sheath cells (BSCs) and mesophyll cells (MCs), respectively. Expression of a beta-glucuronidase (GUS) reporter gene under the control of the PcAat promoter was regulated in an organ-preferential and light-dependent manner in the transgenic rice plants. However, the PmAat promoter drove the GUS expression in all organs we tested without light dependency, and this non-preferential expression pattern was also observed in transgenic rice with introduction of the intact PmAat gene. The expression patterns of the rice counterpart Aat genes to PmAat or PcAat showed that the rice mitochondrial Aat (RmAat1) gene was expressed in all organs tested in a light-independent manner, while expression of the rice cytosolic Aat (RcAat1) gene showed an organ-preferential and light-dependent pattern. Taking these results together, we can generalize that the regulatory system of BSC-specific or light-dependent expression of mitochondrial Aat is not shared between P. miliaceum (C4) and rice (C3) and that the expression of the C4 genes introduced into rice mimics that of their counterpart genes in rice. PMID- 15753103 TI - Differential expression pattern of C4 bundle sheath expression genes in rice, a C3 plant. AB - NADP-malic enzyme (NADP-ME) and phosphoenolpyruvate carboxykinase (PCK) are specifically expressed in bundle sheath cells (BSCs) in NADP-ME-type and PCK-type C4 plants, respectively. Unlike the high activities of these enzymes in the green leaves of C4 plants, their low activities have been detected in the leaves of C3 plants. In order to elucidate the differences in the gene expression system between C3 and C4 plants, we have produced chimeric constructs with the beta glucuronidase (GUS) reporter gene under the control of the maize NADP-Me (ZmMe) or Zoysia japonica Pck (ZjPck) promoter and introduced these constructs into rice. In leaves of transgenic rice, the ZmMe promoter directed GUS expression not only in mesophyll cells (MCs) but also in BSCs and vascular cells, whereas the ZjPck promoter directed GUS expression only in BSCs and vascular cells. Neither the ZjPck nor ZmMe promoters induced GUS expression due to light. In rice leaves, the endogenous NADP-Me (OsMe1) was expressed in MCs, BSCs and vascular cells, whereas the rice Pck (OsPck1) was expressed only in BSCs and vascular cells. Taken together, the results obtained from transgenic rice demonstrate that the expression pattern of ZmMe or ZjPck in transgenic rice was reflected by that of its counterpart gene in rice. PMID- 15753105 TI - Analysis of the function of the photoreceptors phytochrome B and phytochrome D in Nicotiana plumbaginifolia and Arabidopsis thaliana. AB - To investigate the mechanism of phytochrome action in vivo, NtPHYB, AtPHYB and phyD:green fluorescent protein (GFP) were overexpressed in Nicotiana plumbaginifolia and Arabidopsis thaliana. The expression of 35S:NtPHYB:GFP and 35S:AtPHYB:GFP complemented the tobacco hgl2 and Arabidopsis phyB-9 mutations, whereas the 35S:AtPHYD:GFP only rescued the hgl2 mutant. All three fusion proteins are transported into the nucleus in all genetic backgrounds. These data indicate that AtPHYD:GFP is biologically active and functions as the main red light receptor in transgenic tobacco, and establish an experimental system for the functional analysis of this elusive photoreceptor in vivo. PMID- 15753106 TI - ACTIVATOR of Spomin::LUC1/WRINKLED1 of Arabidopsis thaliana transactivates sugar inducible promoters. AB - We isolated an enhancer activation-tagged mutant of Arabidopsis thaliana line sGsL carrying the luciferase (LUC) gene under control of a short sugar-inducible promoter derived from a sweet potato sporamin gene (Spomin) that showed high level expression of LUC under non-inducing conditions. The activator of Spomin::LUC1 (ASML1) gene located downstream of the enhancer encoded an APETALA2 (AP2)-type AP2 domain protein, and this gene was shown recently to be responsible for the wrinkled1 mutation which causes defective accumulation of seed storage oil. Overexpression of ASML1 cDNA in sGsL plants resulted in enhanced expression of not only the LUC reporter but also endogenous sugar-inducible genes including Atbeta-Amy encoding beta-amylase. Transient co-expression of 35S::ASML1 with Spomin::LUC or Atbeta-Amy::LUC reporters in protoplasts resulted in an approximately 10-fold transactivation of LUC expression. This transactivation was lost when the C-terminal acidic region of ASML1 was deleted. Expression of ASML1 was high in reproductive organs, and ASML1 mRNA showed transient accumulation in leaves after treatment with 6% sucrose, whereas it did not respond to abscisic acid. These results suggest that ASML1/WRI1 is a transcriptional activator involved in the activation of a subset of sugar-responsive genes and the control of carbon flow from sucrose import to oil accumulation in developing seeds. PMID- 15753108 TI - Aberrant cell plate formation in the Arabidopsis thaliana microtubule organization 1 mutant. AB - MICROTUBULE ORGANIZATION 1 encodes a microtubule-associated protein in Arabidopsis thaliana but different alleles have contradictory phenotypes. The original mutant mor1 alleles were reported to have disrupted cortical microtubules, swollen organs and normal cytokinesis, whereas other alleles, embryo-lethal gemini pollen 1 (gem1), have defective pollen cytokinesis. To determine whether MOR1 functions generally in cytokinesis, we examined the ultrastructure of cell division in roots of the original mor1-1 allele. Cell plates are misaligned, branched and meandering; the forming cell plates remain partly vesicular, with electron-dense or lamellar content. Phragmoplast microtubules are abundant but organized aberrantly. Thus, MOR1 functions in both phragmoplast and cortical arrays. PMID- 15753107 TI - Plant cell growth and ion flux responses to the streptomycete phytotoxin thaxtomin A: calcium and hydrogen flux patterns revealed by the non-invasive MIFE technique. AB - Thaxtomin A, a key phytotoxin produced by plant pathogenic Streptomyces sp., is implicit in common scab disease expression in potato. Primary targets and modes of action of thaxtomin A toxicity in plant cells are not well understood. In this work, early signalling events associated with thaxtomin A toxicity were studied using the ion-selective microelectrode ion flux estimation (MIFE) technique. Thaxtomin A-induced changes in net ion fluxes were measured across the plasma membrane (PM) of root and pollen tube tissue in Arabidopsis thaliana and tomato. Within a minute after toxin application, a rapid and short-lived Ca2+ influx was observed. Well ahead of the marked inhibition of root growth, a significant shift towards net H+ efflux across the PM occurred in all tissues. Similar to root tissues, thaxtomin A significantly modified ion flux profiles from growing pollen tubes. Thaxtomin A was more effective in young, physiologically active tissues (root elongation zone or pollen tube apex), suggesting a higher density of thaxtomin A-binding sites in these regions. Overall, our data provide the first evidence that thaxtomin A triggers an early signalling cascade, which may be crucial in plant-pathogen interactions. It also suggests a possible interaction between thaxtomin A and PM auxin receptors, as revealed from experiments on the auxin-sensitive ucu2-2/gi2 A. thaliana mutant. PMID- 15753109 TI - Uptake system of silicon in different plant species. AB - The accumulation of silicon (Si) in the shoots varies considerably among plant species, but the mechanism responsible for this variation is poorly understood. The uptake system of Si was investigated in terms of the radial transport from the external solution to the root cortical cells and the release of Si from the cortical cells to the xylem in rice, cucumber, and tomato, which differ greatly in shoot Si concentration. Symplasmic solutions of the root tips were extracted by centrifugation. The concentrations of Si in the root-cell symplast in all species were higher than that in the external solution, although the concentration in rice was 3- and 5-fold higher than that in cucumber and tomato, respectively. A kinetic study showed that the radial transport of Si was mediated by a transporter with a K(m) value of 0.15 mM in all species, but with different V(max) values in the order of rice>cucumber>tomato. In the presence of the metabolic inhibitor 2,4-dinitrophenol, and at low temperature, the Si concentration in the root-cell symplast decreased to a level similar to that of the apoplasmic solution. These results suggest that both transporter-mediated transport and passive diffusion of Si are involved in the radial transport of Si and that the transporter-mediated transport is an energy-dependent process. The Si concentration of xylem sap in rice was 20- and 100-fold higher than that in cucumber and tomato, respectively. In contrast to rice, the Si concentration in the xylem sap was lower than that in the external solution in cucumber and tomato. A kinetic study showed that xylem loading of Si was also mediated by a kind of transporter in rice, but by passive diffusion in cucumber and tomato. These results indicate that a higher density of transporter for radial transport and the presence of a transporter for xylem loading are responsible for the high Si accumulation in rice. PMID- 15753110 TI - Modified sucrose, starch, and ATP levels in two alloplasmic male-sterile lines of B. napus. AB - Alloplasmic lines of Brassica napus with rearranged Arabidopsis thaliana mitochondrial DNA are male sterile and vegetatively altered compared with B. napus cv. Hanna. The CMS lines contain pure nuclear and plastid genomes from B. napus. Cross-sections of leaves revealed elevated starch accumulation and a higher number of chloroplasts per cell area in CMS plants compared with B. napus. The increase in chloroplast density was found to be the result of the smaller mesophyll cells. Sucrose concentration in the leaves of the CMS lines was reduced both in green leaves as well as in leaves from 2 d-etiolated plants. Flower meristem, flower buds, and leaves from green and 2 d-etiolated plants were analysed for ATP and ADP contents. All CMS plant tissues, except for green leaves, possessed lower ATP levels than B. napus. The results indicate that the reduced availability of energy, i.e. ATP and sucrose in the CMS plants, limits plant growth. This is supported by the reduced levels of two D-type cyclin transcripts and the reduced capacity of the CMS plants to recover after etiolation. PMID- 15753111 TI - Isolation of phosphorylated and dephosphorylated forms of the CP43 internal antenna of photosystem II in Hordeum vulgare L. AB - As a consequence of variation in environmental factors, light being the most important one, a number of photosystem II polypeptides may be reversibly phosphorylated by thylakoid-bound kinase(s). Among them, the reaction centre D1 and D2 polypeptides, the PsbH subunit, and the inner antenna CP43. Here, the separation of two forms of CP43 by high-resolution denaturing polyacrylamide gel electrophoresis is reported. By means of immunoblotting with antibody to phosphothreonine-containing proteins and authentic CP43 and limited proteolysis, these two bands could be identified as the phosphorylated and dephosphorylated forms of CP43. Using non-denaturing isoelectrofocusing, a chromatographically derived CP43-enriched fraction could be resolved into three different native forms of CP43. Among them, one was found to be a phosphorylated form, whereas the other two were dephosphorylated forms of the protein. With respect to other methods, the procedure described here allows the isolation, for the first time, of a fully homogeneous population of this chlorophyll-protein complex, opening the way to the study of the role of phopshorylation on functional properties of this core antenna protein. PMID- 15753112 TI - The molecular biology of plastid division in higher plants. AB - Plastids are essential plant organelles vital for life on earth, responsible not only for photosynthesis but for many fundamental intermediary metabolic reactions. Plastids are not formed de novo but arise by binary fission from pre existing plastids, and plastid division therefore represents an important process for the maintenance of appropriate plastid populations in plant cells. Plastid division comprises an elaborate pathway of co-ordinated events which include division machinery assembly at the division site, the constriction of envelope membranes, membrane fusion and, ultimately, the separation of the two new organelles. Because of their prokaryotic origin bacterial cell division has been successfully used as a paradigm for plastid division. This has resulted in the identification of the key plastid division components FtsZ, MinD, and MinE, as well as novel proteins with similarities to prokaryotic cell division proteins. Through a combination of approaches involving molecular genetics, cell biology, and biochemistry, it is now becoming clear that these proteins act in concert during plastid division, exhibiting both similarities and differences compared with their bacterial counterparts. Recent efforts in the cloning of the disrupted loci in several of the accumulation and replication of chloroplasts mutants has further revealed that the division of plastids is controlled by a combination of prokaryote-derived and host eukaryote-derived proteins residing not only in the plastid stroma but also in the cytoplasm. Based on the available data to date, a working model is presented showing the protein components involved in plastid division, their subcellular localization, and their protein interaction properties. PMID- 15753113 TI - Application of Bruchin B to pea pods results in the up-regulation of CYP93C18, a putative isoflavone synthase gene, and an increase in the level of pisatin, an isoflavone phytoalexin. AB - Bruchins, mono and bis (3-hydroxypropanoate) esters of long chain alpha,omega diols, are a recently discovered class of insect elicitors that stimulate cell division and neoplasm formation when applied to pods of peas and certain other legumes. Differential display analysis resulted in the identification of an mRNA whose level was increased by the application of Bruchin B to pea pods. The corresponding amplification product was cloned and sequenced and a full length cDNA sequence was obtained. This cDNA and the gene from which it was derived were assigned the name CYP93C18 based upon sequence similarities to the cytochrome P450 mono-oxygenase CYP93C subfamily, which contains isoflavone synthase genes from legumes. RNA gel blots and quantitative RT-PCR demonstrated that expression of CYP93C18 increased within 8 h of bruchin treatment to a maximum of 100-200 fold of the level in untreated pods, and then declined. The up-regulation of CYP93C18 was followed by an increase in the level of the isoflavone phytoalexin, pisatin. Pisatin was detectable in the bruchin-treated pods after 16 h and reached a maximum between 32 h and 64 h. This, the first report of induction of phytoalexin biosynthesis by an insect elicitor, suggests that Bruchin B not only stimulates neoplasm formation, but also activates other plant defence responses. PMID- 15753114 TI - A randomized trial of low-dose aspirin in the primary prevention of cardiovascular disease in women. AB - BACKGROUND: Randomized trials have shown that low-dose aspirin decreases the risk of a first myocardial infarction in men, with little effect on the risk of ischemic stroke. There are few similar data in women. METHODS: We randomly assigned 39,876 initially healthy women 45 years of age or older to receive 100 mg of aspirin on alternate days or placebo and then monitored them for 10 years for a first major cardiovascular event (i.e., nonfatal myocardial infarction, nonfatal stroke, or death from cardiovascular causes). RESULTS: During follow-up, 477 major cardiovascular events were confirmed in the aspirin group, as compared with 522 in the placebo group, for a nonsignificant reduction in risk with aspirin of 9 percent (relative risk, 0.91; 95 percent confidence interval, 0.80 to 1.03; P=0.13). With regard to individual end points, there was a 17 percent reduction in the risk of stroke in the aspirin group, as compared with the placebo group (relative risk, 0.83; 95 percent confidence interval, 0.69 to 0.99; P=0.04), owing to a 24 percent reduction in the risk of ischemic stroke (relative risk, 0.76; 95 percent confidence interval, 0.63 to 0.93; P=0.009) and a nonsignificant increase in the risk of hemorrhagic stroke (relative risk, 1.24; 95 percent confidence interval, 0.82 to 1.87; P=0.31). As compared with placebo, aspirin had no significant effect on the risk of fatal or nonfatal myocardial infarction (relative risk, 1.02; 95 percent confidence interval, 0.84 to 1.25; P=0.83) or death from cardiovascular causes (relative risk, 0.95; 95 percent confidence interval, 0.74 to 1.22; P=0.68). Gastrointestinal bleeding requiring transfusion was more frequent in the aspirin group than in the placebo group (relative risk, 1.40; 95 percent confidence interval, 1.07 to 1.83; P=0.02). Subgroup analyses showed that aspirin significantly reduced the risk of major cardiovascular events, ischemic stroke, and myocardial infarction among women 65 years of age or older. CONCLUSIONS: In this large, primary-prevention trial among women, aspirin lowered the risk of stroke without affecting the risk of myocardial infarction or death from cardiovascular causes, leading to a nonsignificant finding with respect to the primary end point. PMID- 15753115 TI - The effect of cardiac resynchronization on morbidity and mortality in heart failure. AB - BACKGROUND: Cardiac resynchronization reduces symptoms and improves left ventricular function in many patients with heart failure due to left ventricular systolic dysfunction and cardiac dyssynchrony. We evaluated its effects on morbidity and mortality. METHODS: Patients with New York Heart Association class III or IV heart failure due to left ventricular systolic dysfunction and cardiac dyssynchrony who were receiving standard pharmacologic therapy were randomly assigned to receive medical therapy alone or with cardiac resynchronization. The primary end point was the time to death from any cause or an unplanned hospitalization for a major cardiovascular event. The principal secondary end point was death from any cause. RESULTS: A total of 813 patients were enrolled and followed for a mean of 29.4 months. The primary end point was reached by 159 patients in the cardiac-resynchronization group, as compared with 224 patients in the medical-therapy group (39 percent vs. 55 percent; hazard ratio, 0.63; 95 percent confidence interval, 0.51 to 0.77; P<0.001). There were 82 deaths in the cardiac-resynchronization group, as compared with 120 in the medical-therapy group (20 percent vs. 30 percent; hazard ratio 0.64; 95 percent confidence interval, 0.48 to 0.85; P<0.002). As compared with medical therapy, cardiac resynchronization reduced the interventricular mechanical delay, the end-systolic volume index, and the area of the mitral regurgitant jet; increased the left ventricular ejection fraction; and improved symptoms and the quality of life (P<0.01 for all comparisons). CONCLUSIONS: In patients with heart failure and cardiac dyssynchrony, cardiac resynchronization improves symptoms and the quality of life and reduces complications and the risk of death. These benefits are in addition to those afforded by standard pharmacologic therapy. The implantation of a cardiac-resynchronization device should routinely be considered in such patients. PMID- 15753116 TI - Differences in plastic responses to defoliation due to variation in the timing of treatments for two species of Sesbania (Fabaceae). AB - BACKGROUND AND AIMS: Plastic responses to stress in components of reproduction can have important effects on plant fitness and can vary both within and between species. Responses may also depend on when in the life cycle stress occurs. Here, it is predicted that the timing of initiation of a stress, defoliation, would affect the pattern of plastic responses. These differences should occur because some components of reproduction, such as flower number, are determined earlier in a plant's life than others, such as individual seed mass. METHODS: To test this prediction, 50 % artificial defoliation treatments were initiated at four different times for Sesbania macrocarpa and S. vesicaria. Responses were measured in plant size, number of flowers, number of flowers/plant size, fruit set, number of seeds per fruit, individual seed mass and total seed mass per plant. KEY RESULTS: For S. vesicaria, changes in the timing of stress changed the severity, but not the pattern of response. For S. macrocarpa, plastic responses to defoliation varied strikingly between early and late treatments. Late treatments resulted in over-compensation in this species. Sesbania macrocarpa was generally more plastic than S. vesicaria and the species showed opposite responses for most components of reproduction. CONCLUSIONS: While there were effects of timing of defoliation and differences between species, the nature of these effects did not precisely fit our predictions. Our results suggest that differences in the length and flexibility of the life cycles of the two species allowed for unexpected variation in responses. For example, because flower production continued after the last treatment in S. vesicaria, responses were not constrained to reductions in individual seed mass. PMID- 15753117 TI - Bud morphology and shoot growth dynamics in two species of Mediterranean sub shrubs co-existing in gypsum outcrops. AB - BACKGROUND AND AIMS: Understanding the effects of the environment on the morphology and shoot growth activities of plants is crucial to identifying plant ecological strategies. This study analysed the bud morphology, bud activity, shoot growth dynamics and shoot water content at full hydration (WC(h)) of two species of Mediterranean sub-shrubs, Lepidium subulatum and Linum suffruticosum, co-existing in gypsum outcrops in north-east Spain. METHODS: Sampling was conducted monthly over 2 years in one population per species. Buds were dissected under a stereo-microscope. Shoot growth was measured as the mean increase in shoot length of 15 marked individuals between two consecutive samplings. Bud activity was studied following the variations in the number of leaf primordia shorter than 1 mm and longer than 0.025 mm in the buds. KEY RESULTS: Both species bore naked buds and displayed discontinuous seasonal patterns of shoot growth, leaf primordia formation and WC(h). The number of leaf primordia in the bud peaked before the beginning of shoot expansion. In both species, organogenesis and expansion were uncoupled throughout the year. The time lapse between these two processes varied throughout the year, and was greatest for those elements differentiated in autumn. WC(h) was more closely related to shoot expansion than to organogenesis. CONCLUSIONS: Both species displayed similar bud morphology and similar seasonal patterns of bud and shoot growth, and WC(h) as a result of the strong seasonality of the Mediterranean climate in gypsum outcrops. The beginning of the spring period of expansion of long branches coincided with maximum values of WC(h), while the rest period of summer matched minimum values. These results support the hypothesis that the growth of long branches is strongly related to WC(h). PMID- 15753118 TI - Effects of pollen shortage and self-pollination on seed production of an endangered tree, Magnolia stellata. AB - BACKGROUND AND AIMS: Pollen limitation is a significant determinant of seed production, and can result from both insufficient pollen quantity (pollen shortage) and quality (mainly relating to self-pollination). For animal pollinated tree species with large floral displays, pollen limitation may be determined by a balance between increased pollen quantity due to increased attractiveness for pollinators, countered by increased self-pollination due to increased geitonogamy. The contributions of pollen shortage and self-pollination on seed production were quantitatively examined in the natural pollination of an insect-pollinated, dichogamous, endangered tree, Magnolia stellata, which has a large, showy floral display. METHODS: Manual self- and cross-pollinations were conducted to determine the effects of selfing on seed production. The outcrossing rate was measured using microsatellite analyses of open-pollinated seeds, and the embryo mortality rate caused by self-pollination was indirectly estimated. The frequency of ovule mortality due to pollen shortage was also inferred using the embryo mortality and ovule survival rates from natural pollination. KEY RESULTS: The average fruit set, seed set per fruit, and ovule survival rate per tree from hand cross-pollination were 1.37, 3.15, and 3.34 times higher than those from hand self-pollination, respectively, indicating that self-pollination causes inbreeding depression for fruit and seed set. The multilocus-outcrossing rate (t(m)) was intermediate, 0.632, and the primary selfing rate was 0.657. This indicates that frequent geitonogamous selfing occurs. The ovule mortality rate due to pollen shortage and the embryo mortality rate due to self-pollination were estimated to be 80.8 % and 45.9 %, respectively. CONCLUSIONS: It is concluded that seed production of M. stellata is strongly limited by both pollen shortage and self-pollination. Inefficient beetle-pollination and the automimicry system via asynchronous flowering might be responsible for the high level of pollen shortage and frequent geitonogamy. This is despite a large, showy floral display and the dichogamous system of the species. PMID- 15753120 TI - Identification of phosphopeptides by MALDI Q-TOF MS in positive and negative ion modes after methyl esterification. AB - We have developed an efficient, sensitive, and specific method for the detection of phosphopeptides present in peptide mixtures by MALDI Q-TOF mass spectrometry. Use of the MALDI Q-TOF enables selection of phosphopeptides and characterization by CID of the phosphopeptides performed on the same sample spot. However, this type of experiment has been limited by low ionization efficiency of phosphopeptides in positive ion mode while selecting precursor ions of phosphopeptides. Our method entails neutralizing negative charges on acidic groups of nonphosphorylated peptides by methyl esterification before mass spectrometry in positive and negative ion modes. Methyl esterification significantly increases the relative signal intensity generated by phosphopeptides in negative ion mode compared with positive ion mode and greatly increases selectivity for phosphopeptides by suppressing the signal intensity generated by acidic peptides in negative ion mode. We used the method to identify 12 phosphopeptides containing 22 phosphorylation sites from low femtomolar amounts of a tryptic digest of beta-casein and alpha-s-casein. We also identified 10 phosphopeptides containing five phosphorylation sites from an in-gel tryptic digest of 100 fmol of an in vitro autophosphorylated fibroblast growth factor receptor kinase domain and an additional phosphopeptide containing another phosphorylation site when 500 fmol of the digest was examined. The results demonstrate that the method is a fast, robust, and sensitive means of characterizing phosphopeptides present in low abundance mixtures of phosphorylated and nonphosphorylated peptides. PMID- 15753119 TI - Ethylene signal transduction. AB - BACKGROUND: The phytohormone ethylene is a key regulator of plant growth and development. Components of the pathway for ethylene signal transduction were identified by genetic approaches in Arabidopsis and have now been shown to function in agronomically important plants as well. SCOPE: This review focuses on recent advances in our knowledge on ethylene signal transduction, in particular on recently proposed components of the pathway, on the interaction between the pathway components and on the roles of transcriptional and post-transcriptional regulation in ethylene signalling. CONCLUSIONS: Data indicate that the site of ethylene perception is at the endoplasmic reticulum and point to the importance of protein complexes in mediating the initial steps in ethylene signal transduction. The expression level of pathway components is regulated by both transcriptional and post-transcriptional mechanisms, degradation of the transcription factor EIN3 being a primary means by which the sensitivity of plants to ethylene is regulated. EIN3 also represents a control point for cross talk with other signalling pathways, as exemplified by the effects of glucose upon its expression level. Amplification of the initial ethylene signal is likely to play a significant role in signal transduction and several mechanisms exist by which this may occur based on properties of known pathway components. Signal output from the pathway is mediated in part by carefully orchestrated changes in gene expression, the breadth of these changes now becoming clear through expression analysis using microarrays. PMID- 15753122 TI - Proteomic analysis of primary cultures of human adipose-derived stem cells: modulation by Adipogenesis. AB - Adipogenesis plays a critical role in energy metabolism and is a contributing factor to the obesity epidemic. This study examined the proteome of primary cultures of human adipose-derived adult stem (ADAS) cells as an in vitro model of adipogenesis. Protein lysates obtained from four individual donors were compared before and after adipocyte differentiation by two-dimensional gel electrophoresis and tandem mass spectroscopy. Over 170 individual protein features in the undifferentiated adipose-derived adult stem cells were identified. Following adipogenesis, over 40 proteins were up-regulated by > or = 2-fold, whereas 13 showed a > or = 3-fold reduction. The majority of the modulated proteins belonged to the following functional categories: cytoskeleton, metabolic, redox, protein degradation, and heat shock protein/chaperones. Additional immunoblot analysis documented the induction of four individual heat shock proteins and confirmed the presence of the heat shock protein 27 phosphoserine 82 isoform, as predicted by the proteomic analysis, as well as the crystallin alpha phosphorylated isoforms. These findings suggest that the heat shock protein family proteome warrants further investigation with respect to the etiology of obesity and type 2 diabetes. PMID- 15753121 TI - Quantitative proteome analysis of human plasma following in vivo lipopolysaccharide administration using 16O/18O labeling and the accurate mass and time tag approach. AB - Identification of novel diagnostic or therapeutic biomarkers from human blood plasma would benefit significantly from quantitative measurements of the proteome constituents over a range of physiological conditions. Herein we describe an initial demonstration of proteome-wide quantitative analysis of human plasma. The approach utilizes postdigestion trypsin-catalyzed 16O/18O peptide labeling, two dimensional LC-FTICR mass spectrometry, and the accurate mass and time (AMT) tag strategy to identify and quantify peptides/proteins from complex samples. A peptide accurate mass and LC elution time AMT tag data base was initially generated using MS/MS following extensive multidimensional LC separations to provide the basis for subsequent peptide identifications. The AMT tag data base contains >8,000 putative identified peptides, providing 938 confident plasma protein identifications. The quantitative approach was applied without depletion of high abundance proteins for comparative analyses of plasma samples from an individual prior to and 9 h after lipopolysaccharide (LPS) administration. Accurate quantification of changes in protein abundance was demonstrated by both 1:1 labeling of control plasma and the comparison between the plasma samples following LPS administration. A total of 429 distinct plasma proteins were quantified from the comparative analyses, and the protein abundances for 25 proteins, including several known inflammatory response mediators, were observed to change significantly following LPS administration. PMID- 15753123 TI - The high mobility group transcription factor Sox8 is a negative regulator of osteoblast differentiation. AB - Bone remodeling is an important physiologic process that is required to maintain a constant bone mass. This is achieved through a balanced activity of bone resorbing osteoclasts and bone-forming osteoblasts. In this study, we identify the high mobility group transcription factor Sox8 as a physiologic regulator of bone formation. Sox8-deficient mice display a low bone mass phenotype that is caused by a precocious osteoblast differentiation. Accordingly, primary osteoblasts derived from these mice show an accelerated mineralization ex vivo and a premature expression of osteoblast differentiation markers. To confirm the function of Sox8 as a negative regulator of osteoblast differentiation we generated transgenic mice that express Sox8 under the control of an osteoblast specific Col1a1 promoter fragment. These mice display a severely impaired bone formation that can be explained by a strongly reduced expression of runt-related transcription factor 2, a gene encoding a transcription factor required for osteoblast differentiation. Together, these data demonstrate a novel function of Sox8, whose tightly controlled expression is critical for bone formation. PMID- 15753124 TI - Akt2 phosphorylates Synip to regulate docking and fusion of GLUT4-containing vesicles. AB - We have identified an unusual potential dual Akt/protein kinase B consensus phosphorylation motif in the protein Synip (RxKxRS(97)xS(99)). Surprisingly, serine 97 is not appreciably phosphorylated, whereas serine 99 is only a specific substrate for Akt2 but not Akt1 or Akt3. Although wild-type Synip (WT-Synip) undergoes an insulin-stimulated dissociation from Syntaxin4, the Synip serine 99 to phenylalanine mutant (S99F-Synip) is resistant to Akt2 phosphorylation and fails to display insulin-stimulated Syntaxin4 dissociation. Furthermore, overexpression of WT-Synip in 3T3L1 adipocytes had no effect on insulin stimulated recruitment of glucose transporter 4 (GLUT4) to the plasma membrane, whereas overexpression of S99F-Synip functioned in a dominant-interfering manner by preventing insulin-stimulated GLUT4 recruitment and plasma membrane fusion. These data demonstrate that insulin activation of Akt2 specifically regulates the docking/fusion step of GLUT4-containing vesicles at the plasma membrane through the regulation of Synip phosphorylation and Synip-Syntaxin4 interaction. PMID- 15753125 TI - Chromatin decondensation in S-phase involves recruitment of Cdk2 by Cdc45 and histone H1 phosphorylation. AB - Cdc45 is required for initiation of DNA replication and fork progression, but its function in these processes remains unknown. We show that targeting Cdc45 to specific chromosomal sites in mammalian cells results in large-scale chromatin decondensation that strongly correlates with histone H1 phosphorylation. Cdk2 is recruited to sites of Cdc45 decondensation, and Cdk2 inhibitors reduce the level of decondensation. Targeting wild-type Cdk2, but not kinase-defective Cdk2, to chromatin is also effective at inducing decondensation involving phospho-H1. Cdc45, Cdk2, Cyclin A, and phospho-H1 associate with chromatin during S-phase, and Cdc45, Cdk2, and an active H1 kinase physically interact. Replicating DNA and phospho-H1 foci colocalize in vivo, and S-phase progression and H1 phosphorylation are directly related and Cdk2 dependent. Because Cdk2 colocalizes with replication foci and H1 regulates higher-order chromatin, we suggest a model in which Cdc45 recruits Cdk2 to replication foci, resulting in H1 phosphorylation, chromatin decondensation, and facilitation of fork progression. PMID- 15753126 TI - TRPV4 channel is involved in the coupling of fluid viscosity changes to epithelial ciliary activity. AB - Autoregulation of the ciliary beat frequency (CBF) has been proposed as the mechanism used by epithelial ciliated cells to maintain the CBF and prevent the collapse of mucociliary transport under conditions of varying mucus viscosity. Despite the relevance of this regulatory response to the pathophysiology of airways and reproductive tract, the underlying cellular and molecular aspects remain unknown. Hamster oviductal ciliated cells express the transient receptor potential vanilloid 4 (TRPV4) channel, which is activated by increased viscous load involving a phospholipase A(2)-dependent pathway. TRPV4-transfected HeLa cells also increased their cationic currents in response to high viscous load. This mechanical activation is prevented in native ciliated cells loaded with a TRPV4 antibody. Application of the TRPV4 synthetic ligand 4alpha-phorbol 12,13 didecanoate increased cationic currents, intracellular Ca(2+), and the CBF in the absence of a viscous load. Therefore, TRPV4 emerges as a candidate to participate in the coupling of fluid viscosity changes to the generation of the Ca(2+) signal required for the autoregulation of CBF. PMID- 15753128 TI - ROCK-I regulates closure of the eyelids and ventral body wall by inducing assembly of actomyosin bundles. AB - Rho-associated kinase (ROCK) I mediates signaling from Rho to the actin cytoskeleton. To investigate the in vivo functions of ROCK-I, we generated ROCK-I deficient mice. Loss of ROCK-I resulted in failure of eyelid closure and closure of the ventral body wall, which gave rise to the eyes open at birth and omphalocele phenotypes in neonates. Most ROCK-I(-/-) mice died soon after birth as a result of cannibalization of the omphalocele by the mother. Actin cables that encircle the eye in the epithelial cells of the eyelid were disorganized and accumulation of filamentous actin at the umbilical ring was impaired, with loss of phosphorylation of the myosin regulatory light chain (MLC) at both sites, in ROCK-I(-/-) embryos. Stress fiber formation and MLC phosphorylation induced by EGF were also attenuated in primary keratinocytes from ROCK-I(-/-) mice. These results suggest that ROCK-I regulates closure of the eyelids and ventral body wall through organization of actomyosin bundles. PMID- 15753129 TI - Vitamin E trafficking. AB - The alpha-tocopherol transfer protein (alpha-TTP) is required to prevent vitamin E deficiency in humans and in alpha-TTP null mice. Whereas alpha-TTP is not required to facilitate intestinal absorption of vitamin E, it is required to maintain normal alpha-tocopherol concentrations in plasma and extrahepatic tissues. alpha-Tocopherol secretion from the liver in very low density lipoproteins (VLDLs) is impaired in humans with a defect in the alpha-TTP gene. In perfusions of isolated cynomolgus monkey livers, VLDLs were preferentially enriched in RRR-alpha-tocopherol. The mechanism by which alpha-TTP incorporates alpha-tocopherol into nascent VLDLs is the topic of this report. VLDL assembly is a multistep secretory process that occurs within the membrane compartments of the endoplasmic reticulum and Golgi apparatus. Thus, we postulated that alpha-TTP might transfer alpha-tocopherol onto nascent VLDLs either in the endoplasmic reticulum or in the Golgi apparatus. To test these possibilities, we isolated nascent VLDLs from highly purified RER and Golgi apparatus membrane fractions from livers of rats fed equimolar ratios of RRR- and SRR-alpha-tocopherols labeled with different amounts of deuterium. Although the plasma was enriched in RRR-alpha-tocopherol 14 hours after the dose, no enrichment of nascent VLDL precursors from either of the secretory compartments was detected, indicating that VLDL enrichment with alpha-tocopherol may occur as a post-VLDL secretory process. Therefore, we hypothesize that alpha-TTP may facilitate movement of alpha-tocopherol to the hepatocyte plasma membrane (by unknown mechanisms) where newly secreted, nascent VLDLs could acquire both alpha-tocopherol and unesterified cholesterol while within the space of Disse. Clearly, critical information is lacking in our understanding of the mechanism by which alpha-TTP facilitates the preferential enrichment of VLDLs with alpha-tocopherol. PMID- 15753127 TI - Raf-1 regulates Rho signaling and cell migration. AB - Raf kinases relay signals inducing proliferation, differentiation, and survival. The Raf-1 isoform has been extensively studied as the upstream kinase linking Ras activation to the MEK/ERK module. Recently, however, genetic experiments have shown that Raf-1 plays an essential role in counteracting apoptosis, and that it does so independently of its ability to activate MEK. By conditional gene ablation, we now show that Raf-1 is required for normal wound healing in vivo and for the migration of keratinocytes and fibroblasts in vitro. Raf-1-deficient cells show a symmetric, contracted appearance, characterized by cortical actin bundles and by a disordered vimentin cytoskeleton. These defects are due to the hyperactivity and incorrect localization of the Rho-effector Rok-alpha to the plasma membrane. Raf-1 physically associates with Rok-alpha in wild-type (WT) cells, and reintroduction of either WT or kinase-dead Raf-1 in knockout fibroblasts rescues their defects in shape and migration. Thus, Raf-1 plays an essential, kinase-independent function as a spatial regulator of Rho downstream signaling during migration. PMID- 15753130 TI - Discovery, characterization, and significance of the cytochrome P450 omega hydroxylase pathway of vitamin E catabolism. AB - Tocopherols are known to undergo metabolism to phytyl chain-shortened metabolites excreted in urine. We sought to characterize the pathway, including associated enzymes, involved in this biotransformation. We previously found that human hepatoblastoma (HepG2) cultures metabolized tocopherols to their corresponding short-chain carboxychromanols. Putative metabolites of gamma-tocopherol that contained intact chromanol moieties were structurally identified using HepG2 cultures and electron impact gas chromatography-mass spectrometry. A microsomal assay for synthesis of the initial omega-oxidation metabolites was developed and used to screen several recombinant human liver cytochrome P450 isozymes for omega hydroxylase activity. Seven metabolites of gamma-tocopherol were identified in HepG2 cultures, including 13'-hydroxy-gamma-TOH and all six carboxychromanols predicted by sequential omega-oxidation truncation. Rat and human liver microsomes catalyzed synthesis of 13'-OH- and 13'-COOH-gamma-TOH, but not other metabolites, in the presence of NADPH. Inclusion of NAD favored synthesis of the 13'-COOH metabolite. Recombinant CYP4F2, but not other major human liver CYP isoforms (including CYP3A4 and 3A7), exhibited tocopherol-omega-hydroxylase activity. Liver microsomes and recombinant CYP4F2 both exhibited substrate preference for gamma-TOH over alpha-TOH, and recent studies show that tocotrienols are catabolized more extensively than the corresponding tocopherols. Comparative rates of omega-oxidation of tocochromanols in hepatocytes are inversely related to biopotency and directly related to cytotoxicity of these substances in macrophages. The liver contains a cytochrome P450-mediated pathway that preferentially catabolizes "non-alpha" tocochromanols to excretable metabolites. This metabolic pathway appears central to the optimization of tissue tocochromanol status. PMID- 15753132 TI - The effect of age on vitamin E status, metabolism, and function: metabolism as assessed by labeled tocopherols. AB - The effects of age on vitamin E metabolism were studied in 97 healthy 20-75-year old male nonsmoking Austrian volunteers of the VITAGE project. After a single oral intake of 30 mg d(6)-RRR-alpha- and d(2)-RRR-gamma-tocopheryl acetate, blood and 24-hour urine was collected. Deuterated tocopherols in plasma and deuterated urinary metabolites were analyzed by GC-MS. A first evaluation revealed a similar uptake of d(6)-alpha- and d(2)-gamma-tocopherol during the first 6 hours, and then d(2)-gamma-tocopherol started to decrease. Urinary d(2)-gamma- carboxyethyl hydroxychroman metabolites (CEHCs) exceeded those of d(6)-alpha-CEHCs by about 10 times. There was no effect of age. Thus, there might be no need for a higher vitamin E intake for healthy elderly nonsmoking men. PMID- 15753131 TI - Inter- and intra-individual vitamin E uptake in healthy subjects is highly repeatable across a wide supplementation dose range. AB - Vitamin E uptake after supplementation varies widely in the healthy population, and preliminary studies have indicated that individual responses are relatively stable over periods in excess of 1 year. This phenotypic stability suggests a genetic basis to this observed variation. To examine this issue further, we examined the repeatability of both baseline plasma alpha-tocopherol and urinary alpha-tocopherol metabolite concentrations, as well as individual responses of these parameters after vitamin E supplementation. In the first study, 65 subjects (33 males, 32 females, aged 30.7 +/- 7.4 years) provided three plasma and urine samples for alpha-tocopherol and metabolite analysis with each collection separated by at least 2 weeks. Plasma alpha-tocopherol concentrations were found to be highly repeatable over this short interval (intra-class correlation coefficient [ICC] = 0.85), although the association deteriorated once values were corrected for plasma cholesterol (ICC = 0.64). Similarly, urinary alpha tocopherol metabolites 2(2'-carboxyethyl)-6-hydroxychroman acid (alpha-CEHC) and quinone lactone (QL) concentration were found to display a moderate degree of intra-subject repeatability: ICC = 0.65 and 0.58, respectively. In a second study, plasma alpha-tocopherol and urinary metabolite responses were investigated in 18 healthy, nonsmoking subjects (12 males, 6 females, aged 33.1 +/- 9.1 years) after successive 6-week periods of vitamin E (RRR-alpha-tocopherol acetate) supplementation at 15, 100, 200, and 400 mg/day. Plasma and urine samples were obtained on days 0, 7, 14, 21, and 28 (7 days after the final supplement) of each dosing period and the strength of the underlying association between responses determined using Kendall's tau_b test. Individual plasma alpha-tocopherol responses at the 100, 200, and 400 mg/day doses were found to be highly associated: tau, 0.51, P = 0.02 [100 vs. 200] and tau, 0.49, P = 0.03 [100 vs. 400] and tau, 0.56, P = 0.005 [200 vs. 400]. Together these data support the contention that alpha-tocopherol uptake is a stable individual phenotype under genetic regulation. PMID- 15753133 TI - Molecular mechanisms of vitamin E transport. AB - Vitamin E is the most important lipid-soluble antioxidant in humans. Specific tocopherol-binding proteins favor the retention of the most potent vitamin E homologue, RRR-alpha-tocopherol (RRR-alpha-T) in man. The crystal structures of both the ligand-charged and the apo-forms of human alpha-tocopherol transfer protein (alpha-TTP) and of human supernatant protein factor (SPF) have been solved. The renewed interest in the biological function of tocopherol binders is based on the discovery of ataxia with vitamin E deficiency, a neurological disorder that is caused by genetic defects of the alpha-TTP gene and/or vitamin E deficiency. The analysis of the crystal structure of alpha-TTP provides the molecular basis of vitamin E retention in man. SPF has been reported to enhance cholesterol biosynthesis by facilitating the conversion of squalene to lanosterol. Nevertheless, the physiological role of SPF as well as its ligand specificity is not known. Investigations on the substrate specificity of SPF have uncovered binding of RRR-alpha-tocopherylquinone (RRR-alpha-TQ). RRR-alpha-TQ represents the major physiological oxidation product of RRR-alpha-T. The three dimensional overlay of the ligand-charged structures of SPF and alpha-TTP indicates that ligand specificity in both proteins is mostly modulated by side chain variations rather than by the backbone. Recent reports point towards the in vivo reduction of RRR-alpha-TQ to RRR-alpha-TQH(2) and its protective role in low density lipoprotein oxidation. On the basis of these reports, it is proposed that SPF may enhance cholesterol biosynthesis indirectly by mediating the transfer of RRR-alpha-TQ to low-density lipoprotein, thus reducing oxidation of low-density lipoprotein and its subsequent cellular uptake by scavenger receptors. PMID- 15753134 TI - Physiological factors influencing vitamin E biokinetics. AB - Limited information is available on factors that can influence vitamin E bioavailability. In several studies we have investigated the influence of dietary, biochemical, and genetic factors on vitamin E biokinetics. In these studies, subjects ingested a capsule containing 150 mg deuterated RRR-alpha tocopheryl acetate, blood was taken up to 48 hr, and tocopherols were analyzed by liquid chromatography and mass spectroscopy. There was significantly greater plasma-labeled alpha-tocopherol concentrations when the capsule was consumed with a high-fat meal (17.5 g) versus a low-fat meal (2.7 g), and there was also a difference between a high-fat toast and butter and a cereal with full-fat milk meal (both 17.5 g fat), indicating that both the amount of fat and food matrix is important for vitamin E absorption. Dyslipidemic subjects displayed a reduced plasma uptake of newly absorbed alpha-tocopherol, and differences were also apparent in individual lipoproteins. A decreased uptake of labeled alpha tocopherol was also observed in erythrocytes, platelets, and lymphocytes of dyslipidemics. Following vitamin E supplementation (400 mg/day, 4 weeks), the uptake of newly absorbed alpha-tocopherol was decreased, presumably because of saturation of alpha-tocopherol transfer protein. We also found that apoE3 subjects displayed a considerably reduced uptake of newly absorbed labeled alpha tocopherol compared to apoE4 subjects, which may be a consequence of the reduced low-density lipoprotein catabolic rate in these subjects. Taken together, these data show that several physiological factors influence the uptake of newly absorbed alpha-tocopherol, and that this is an important consideration in the design of future vitamin E supplementation studies. PMID- 15753135 TI - Alpha-Tocopherol and endothelial nitric oxide synthesis. AB - Nitric oxide (NO), a central regulator of vascular tone and homeostasis, is generated upon activation of endothelial NO synthase (eNOS), which is mediated by an increase of intracellular calcium and/or by eNOS phosphorylation. A reduction of NO bioavailability leads to endothelial dysfunction that has been shown to be improved by alpha-tocopherol in certain conditions. The underlying mechanisms, however, are not completely clarified. The present study was performed to investigate whether alpha-tocopherol is able to affect endothelial NO synthesis. The formation of NO was measured in human umbilical vein endothelial cells using citrulline (coproduct) and cGMP (product of the NO-activated soluble guanylate cyclase) as indicator molecules. alpha-Tocopherol (10-200 microM, 24 hr) increased ionomycin-induced citrulline and cGMP formation in intact cells in a concentration-dependent manner. In parallel, ionomycin-stimulated phosphorylation of eNOS at serine 1177, known to support enzyme activation, was increased by alpha-tocopherol, suggesting that this was the mechanism responsible for enhanced NO formation. The effect of alpha-tocopherol was dependent on its hydrophobic structure because it was mimicked by gamma-tocopherol but not by trolox, a hydrophilic derivative of alpha-tocopherol. Coincubation with ascorbic acid (100 microM, 24 hr) amplified the effects of alpha-tocopherol on eNOS phosphorylation and NO formation, which is possibly related to the regeneration of oxidized alpha tocopherol by ascorbate. Our data suggest that vasoprotective effects of alpha tocopherol in vivo may be related to an increase of NO formation. The effect of alpha-tocopherol seems to be dependent on tissue saturation with ascorbic acid, and both vitamins may act synergistically to provide optimal conditions for endothelial NO formation. PMID- 15753136 TI - Vitamin E mediates cell signaling and regulation of gene expression. AB - alpha-Tocopherol modulates two major signal transduction pathways centered on protein kinase C and phosphatidylinositol 3-kinase. Changes in the activity of these key kinases are associated with changes in cell proliferation, platelet aggregation, and NADPH-oxidase activation. Several genes are also regulated by tocopherols partly because of the effects of tocopherol on these two kinases, but also independently of them. These genes can be divided in five groups: Group 1. Genes that are involved in the uptake and degradation of tocopherols: alpha tocopherol transfer protein, cytochrome P450 (CYP3A), gamma-glutamyl-cysteine synthetase heavy subunit, and glutathione-S-transferase. Group 2. Genes that are implicated with lipid uptake and atherosclerosis: CD36, SR-BI, and SR-AI/II. Group 3. Genes that are involved in the modulation of extracellular proteins: tropomyosin, collagen-alpha-1, MMP-1, MMP-19, and connective tissue growth factor. Group 4. Genes that are connected to adhesion and inflammation: E selectin, ICAM-1 integrins, glycoprotein IIb, IL-2, IL-4, IL-1b, and transforming growth factor-beta (TGF-beta). Group 5. Genes implicated in cell signaling and cell cycle regulation: PPAR-gamma, cyclin D1, cyclin E, Bcl2-L1, p27, CD95 (APO 1/Fas ligand), and 5a-steroid reductase type 1. The transcription of p27, Bcl2, alpha-tocopherol transfer protein, cytochrome P450 (CYP3A), gamma-glutamyl cysteine sythetase heavy subunit, tropomyosin, IL-2, and CTGF appears to be upregulated by one or more tocopherols. All the other listed genes are downregulated. Gene regulation by tocopherols has been associated with protein kinase C because of its deactivation by alpha-tocopherol and its contribution in the regulation of a number of transcription factors (NF-kappaB, AP1). A direct participation of the pregnane X receptor (PXR) / retinoid X receptor (RXR) has been also shown. The antioxidant-responsive element (ARE) and the TGF-beta responsive element (TGF-beta-RE) appear in some cases to be implicated as well. PMID- 15753137 TI - Vitamin E and gene expression in immune cells. AB - Aging is associated with dysregulation of immune cells, particularly T cells. Previous studies indicated that vitamin E improves T cell function, in part by a direct effect on T cells. We studied gene expression profile of T cells to better understand the underlying mechanisms of aging- and vitamin E-induced changes in T cell function. Young and old C57BL mice were fed diets containing 30 (control) or 500 (E) ppm of vitamin E for 4 weeks. T cells were purified from splenocytes by negative selection using magnetic beads (anti-Mac-1 and anti-MHC class II), then cultured with media or stimulated with anti-CD3 and anti-CD28. Gene expression profile was assessed using microarray analysis. Genes showing more than two-fold changes, P < 0.05 by ANOVA, and with at least one present call were selected. Aging had significant effects on genes involved in signal transduction, transcriptional regulation, and apoptosis pathways in T cells, while vitamin E had a significant effect on genes associated with the regulation of cell cycle. PMID- 15753138 TI - Modulation of hepatic gene expression by alpha-tocopherol in cultured cells and in vivo. AB - To obtain a comprehensive understanding of the molecular mechanisms of action of vitamin E (VE), global gene expression profiles using DNA arrays in rat liver and hepatocellular liver carcinoma cells (HepG2) were obtained. For the analysis of short-term (49 days) and long-term (290 days) VE deficiency, rats were fed semisynthetic diets either supplemented with or deficient in VE. In addition, HepG2 cells were treated with VE concentrations comparable to those that were achieved in the in vivo experiment. Differential gene expression in rat liver and that in HepG2 cells were measured by DNA arrays comprising up to 7,000 genes. Dietary VE deficiency over a 7-week period did not induce any significant changes in the expression profile among the genes evaluated. However, long-term VE deficiency upregulated coagulation factor IX (FIX), 5-alpha-steroid reductase type 1, and CD36 mRNA levels. Furthermore, VE deficiency resulted in a significant downregulation of hepatic gamma-glutamyl-cysteinyl synthetase, the rate-limiting enzyme of glutathione synthesis. According to the rat experiment, VE supplementation changed coagulation factor IX and CD36 expression in HepG2 cells; thus, in vivo data could be partly confirmed with the in vitro model. Overall, the current studies reveal that dietary VE has important long-term effects on liver gene expression with potential downstream effects on extrahepatic tissues. PMID- 15753139 TI - Alpha-tocopherol transfer protein deficiency in mice causes multi-organ deregulation of gene networks and behavioral deficits with age. AB - Functions of alpha-tocopherol (alpha-T) in vivo, other than those for fertility in females, are intensely debated. The discovery of alpha-T deficiency in patients with ataxia (AVED) followed by the identification of mutations in the gene encoding alpha-tocopherol transfer protein (TTP) in AVED patients demonstrates an essential role of alpha-T and TTP for normal neurological function. alpha-T molecular targets that account for alpha-T-sensitive neurological dysfunction remain to be discovered. We have used high-density oligonucleotide arrays to search for putative alpha-T-sensitive genes in the CNS and other tissues in an in vivo model of alpha-T deficiency imposed at birth by the deletion of the TTP gene in mice. Repression of genes affecting synaptic function and myelination and induction of genes for neurodegeneration in the motor cortex of alpha-T-deficient mice were identified. The expression of retinoic acid-related orphan receptor alpha (ROR-alpha) was repressed in the cortex and adrenal glands of TTP-deficient mice. Deficiency of ROR-alpha causes ataxia in mice and may account for ataxia in AVED patients. These observations suggest that some of the actions of alpha-T are mediated by the transcription factor ROR-alpha. The behavior of young TTP-null mice was essentially normal, but older mice showed inactivity, ataxia, and memory dysfunction. mRNA profiles of old alpha-T-deficient cerebral cortices are compatible with repressed activity of oligodendrocytes and astrocytes. In conclusion, gene-expression profiling studies have identified novel alpha-T-modulated genes and cells in the CNS that may be causatively linked with delayed neurodegeneration and age-related decline in behavioral repertoires. PMID- 15753140 TI - Tocotrienol: the natural vitamin E to defend the nervous system? AB - Vitamin E is essential for normal neurological function. It is the major lipid soluble, chain-breaking antioxidant in the body, protecting the integrity of membranes by inhibiting lipid peroxidation. Mostly on the basis of symptoms of primary vitamin E deficiency, it has been demonstrated that vitamin E has a central role in maintaining neurological structure and function. Orally supplemented vitamin E reaches the cerebrospinal fluid and brain. Vitamin E is a generic term for all tocopherols and their derivatives having the biological activity of RRR-alpha-tocopherol, the naturally occurring stereoisomer compounds with vitamin E activity. In nature, eight substances have been found to have vitamin E activity: alpha-, beta-, gamma- and delta-tocopherol; and alpha-, beta , gamma- and delta-tocotrienol. Often, the term vitamin E is synonymously used with alpha-tocopherol. Tocotrienols, formerly known as zeta, , or eta tocopherols, are similar to tocopherols except that they have an isoprenoid tail with three unsaturation points instead of a saturated phytyl tail. Although tocopherols are predominantly found in corn, soybean, and olive oils, tocotrienols are particularly rich in palm, rice bran, and barley oils. Tocotrienols possess powerful antioxidant, anticancer, and cholesterol-lowering properties. Recently, we have observed that alpha-tocotrienol is multi-fold more potent than alpha-tocopherol in protecting HT4 and primary neuronal cells against toxicity induced by glutamate as well as by a number of other toxins. At nanomolar concentration, tocotrienol, but not tocopherol, completely protected neurons by an antioxidant-independent mechanism. Our current work identifies two major targets of tocotrienol in the neuron: c-Src kinase and 12-lipoxygenase. Dietary supplementation studies have established that tocotrienol, fed orally, does reach the brain. The current findings point towards tocotrienol as a potent neuroprotective form of natural vitamin E. PMID- 15753141 TI - Tocotrienol-rich fraction from palm oil and gene expression in human breast cancer cells. AB - Vitamin E is important not only for its cellular antioxidant and lipid-lowering properties, but also as an antiproliferating agent. It has also been shown to contribute to immunoregulation, antibody production, and resistance to implanted tumors. It has recently been shown that tocotrienols are the components of vitamin E responsible for growth inhibition in human breast cancer cells in vitro as well as in vivo through estrogen-independent mechanisms. Although tocotrienols act on cell proliferation in a dose-dependent manner and can induce programmed cell death, no specific gene regulation has yet been identified. In order to investigate the molecular basis of the effect of a tocotrienol-rich fraction (TRF) from palm oil, we performed a cDNA array analysis of cancer-related gene expression in estrogen-dependent (MCF-7) and estrogen-independent (MDA-MB-231) human breast cancer cells. The human breast cancer cells were incubated with or without 8 mug/mL of tocotrienols for 72 h. RNA was subsequently extracted and subjected to reverse transcription before being hybridized onto cancer arrays. Tocotrienol supplementation modulated significantly 46 out of 1200 genes in MDA MB-231 cells. In MCF-7 cells, tocotrienol administration was associated with a lower number of affected genes. Interestingly, only three were affected in a similar fashion in both cell lines: c-myc binding protein MM-1, 23-kDa highly basic protein, and interferon-inducible protein 9-27 (IFITM-1). These proteins are most likely involved in the cell cycle and can exert inhibitory effects on cell growth and differentiation of the tumor cell lines. These data suggest that tocotrienols are able to affect cell homeostasis, possibly independent of their antioxidant activity. PMID- 15753142 TI - Vitamin E and the oxidative stress of exercise. AB - There is clear evidence that contracting skeletal muscle generates a complex set of reactive oxygen and nitrogen species and that the pattern and magnitude of this generation is influenced by the type and frequency of the muscle contraction protocol. The functions of these species in exercising organisms are still unclear although data have been presented indicating that they play a role in contraction-induced muscle damage and/or in signaling adaptive responses to contractions. Vitamin E has been claimed to exert a regulatory effect on the actions of contraction-induced oxidants for a considerable time, although evidence for any specific role in this area is lacking. A review of studies in this area suggests that vitamin E supplements are unlikely to reliably reduce the severity of contraction-induced muscle damage but, in contrast, appear capable of modulating redox-regulated adaptive responses to contractions. Full evaluation of the roles of oxidants and antioxidants such as vitamin E in responses of muscle to contractions should enable the manipulation of these processes with potential beneficial effects on maintenance of optimal muscle function. PMID- 15753143 TI - Effect of vitamin E on gene expression changes in diet-related carcinogenesis. AB - Colorectal cancer (CRC) is responsible for the second highest associated mortality in Western Europe and the United States. Approximately 95% of CRC is sporadic and believed to involve environmental agents and chronic inflammation as causal elements. Several recent studies have suggested a link with diet, in particular, red meat, dietary fats, and low consumption of vegetables. Lipid peroxidation and arachidonic acid metabolism have specifically been implicated in genotoxicity, tumor initiation, and promotion. We have examined the global gene expression profiles (Affymetrix; HU133A) of differentiated vs. undifferentiated colonocytes (CRL-1807), with and without vitamin E supplementation, while undergoing a lipid peroxidative stress. Malondialdehyde and hydroxynonenal, generated by heating a mixture of linoleic and linolenic acid, caused DNA adduct formation identified by immunofluoresence. We also observed a decreased ability for vitamin E to upregulate detoxifying enzymes against free-radical peroxidation, with the exception of mitochondrial superoxide dismutase in undifferentiated cells. However, there was an increased ability in undifferentiated, rather than in differentiated, colonic cells to detect DNA damage, initiate cytostasis, and then effect subsequent DNA repair and apoptosis, in the presence of vitamin E. The expression profile implies less genotoxic stress is experienced in vitamin E-supplemented colonocytes, particularly undifferentiated cells, and points to a mechanism by which dietary supplementation may prevent genotoxic damage and subsequent carcinogenic events in the colon, by both antioxidant and non-antioxidant-related mechanisms. PMID- 15753144 TI - Oral supplementation with all-Rac- and RRR-alpha-tocopherol increases vitamin E levels in human sebum after a latency period of 14-21 days. AB - In human skin, highest alpha-tocopherol levels are found in facial sebum. We hypothesized that the bioavailability of vitamin E in human skin is, at least in part, dependent on sebaceous gland secretion. To test this, 24 volunteers were subjected to a randomized daily supplementation with either 400 mg RRR-alpha tocopheryl acetate (RRR-alpha-toc) or 400 mg all-rac-alpha-tocopheryl acetate (all-rac-alpha-toc) for 14 days. Fasting blood samples, facial sebum samples, and lower-arm skin-surface lipids (SSL) were taken at time-points between 0-21 days. Samples were analyzed by HPLC for alpha-tocopherol and squalene concentrations. Increased serum alpha-tocopherol levels were detectable as early as 12 h after supplementation of RRR-alpha-toc or all-rac-alpha-toc and peaked on day 7. No significant changes were observed in lower-arm SSL. Remarkably, while unchanged until day 14, alpha-tocopherol sebum levels were increased on day 21 in both the RRR-alpha-toc and the all-rac-alpha-toc group by 87% and 92%, respectively. With respect to dietary supplementation of vitamin E and its bioavailability in human skin, these results suggest that (1) sebaceous gland secretion is a relevant delivery mechanism; (2) the bioavailabilities of RRR-alpha-toc and the all-rac alpha-toc are similar; and (3) significant accumulation requires a daily supplementation period of at least 2-3 weeks. PMID- 15753145 TI - Anti-inflammatory effects of alpha-tocopherol. AB - Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in the western world. Its incidence has been increasing lately in the developing countries. Much evidence suggests a major role for inflammation in all phases of atherosclerosis. Cell adhesion molecules, cytokines, chemokines, and monocytes macrophages as well as T lymphocytes play a pivotal role in atherogenesis. C reactive protein (CRP), a downstream marker of inflammation, in addition to being a risk marker for CVD, could contribute to atherosclerosis. Dietary micronutrients with anti-inflammatory properties, specially alpha-tocopherol, may play an important role with regard to the prevention and treatment of CVD. alpha Tocopherol has been shown to have anti-inflammatory effects both in vitro and in vivo. alpha-Tocopherol therapy, especially at high doses, has been shown to decrease release of pro-inflammatory cytokines (such as interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha) and the chemokine interleukin-8, and to decrease adhesion of monocytes to endothelium. In addition, alpha tocopherol has been shown to decrease CRP levels in patients with CVD and having related risk factors for CVD (such as diabetes and smoking). Furthermore, pro inflammatory cytokines and plasminogen activator inhibitor-1 (PAI-1) levels have also been shown to be decreased with alpha-tocopherol supplementation in vivo. In this review, our focus will be on anti-inflammatory effects of alpha-tocopherol reported in in vivo studies. PMID- 15753146 TI - Oxidative stress and antioxidant treatment in diabetes. AB - The many studies on oxidative stress, antioxidant treatment, and diabetic complications have shown that oxidative stress is increased and may accelerate the development of complications through the metabolism of excessive glucose and free fatty acids in diabetic and insulin-resistant states. However, the contribution of oxidative stress to diabetic complications may be tissue specific, especially for microvascular disease that occurs only in diabetic patients but not in individuals with insulin resistance without diabetes, even though both groups suffer from oxidative stress. Although antioxidant treatments can show benefits in animal models of diabetes, negative evidence from large clinical trials suggests that new and more powerful antioxidants need to be studied to demonstrate whether antioxidants can be effective in treating complications. Furthermore, it appears that oxidative stress is only one factor contributing to diabetic complications; thus, antioxidant treatment would most likely be more effective if it were coupled with other treatments for diabetic complications. PMID- 15753147 TI - Vitamin E and respiratory infection in the elderly. AB - Respiratory infections are prevalent in the elderly, resulting in increased morbidity, mortality, and utilization of health care services. Contributing to the increased incidence of infection with age is the well-described decline in immune response, which has been correlated with patterns of illness in the elderly. For example, there are higher morbidity and mortality from cancer, pneumonia, and post-operative complications in those who have diminished, delayed type hypersensitivity skin test responses. Nutritional status is an important determinant of immune function. We have shown in double-blind, placebo-controlled trials that vitamin E supplementation significantly improved immune response, including DTH and response to vaccines. Furthermore, subjects receiving vitamin E in the 6-month trial had a 30% lower incidence of infectious diseases. That study, however, was not powered to demonstrate statistical significance, and the infections were self-reported. To overcome these limitations, we conducted a double-blind, placebo-controlled trial to determine the effect of one-year supplementation with 200 IU/day vitamin E on the incidence and duration of respiratory infections in 617 elderly nursing home residents. The results of this clinical trial show that vitamin E supplementation significantly reduces the incidence rate of common colds and the number of subjects who acquire a cold among elderly nursing home residents. A nonsignificant reduction in the duration of colds was also observed. Because of the high rate and more severe morbidity associated with common colds in this age group, these findings have important implications for the well being of the elderly as well as for the economic burden associated with their care. PMID- 15753149 TI - Selenium and vitamin E cancer prevention trial. AB - Preclinical, epidemiological, and phase III data from randomized, placebo controlled clinical trials suggest that both selenium and vitamin E have potential efficacy in prostate cancer prevention. In vitro evidence suggests that selenium and vitamin E work synergistically to cause cell-cycle arrest, induce caspase-mediated apoptosis, and act as antiandrogens in arresting clonal expansion of nascent tumors. The Selenium and Vitamin E Cancer Prevention Trial (SELECT), sponsored by the National Cancer Institute, is an intergroup Phase III, randomized, double-blind, placebo-controlled, population-based clinical trial designed to test the efficacy of selenium and vitamin E alone and in combination in the prevention of prostate cancer. The study has a 2 x 2 factorial design with a target accrual of 32,400. Eligibility criteria include an age of at least 50 years for African Americans and of at least 55 years for Caucasians; a DRE not suspicious for cancer; a serum PSA no greater than 4 ng/mL; and a normal blood pressure. Randomization will be equally distributed among the four study arms, with intervention consisting of a daily oral dose of study supplement (200 mug l selenomethionine or 400 mg of racemic alpha-tocopheryl) or matched placebo. Study duration is planned for 12 years, with a 5-year uniform accrual period and a minimum of 7 and maximum of 12 years of intervention. The primary endpoint for SELECT is the clinical incidence of prostate cancer as determined by a recommended routine clinical diagnostic work-up, including yearly DRE and serum PSA level. SELECT is the second large-scale study of chemoprevention for prostate cancer. Enrollment began in 2001, with final results anticipated in 2013. PMID- 15753148 TI - Tocopherols and the treatment of colon cancer. AB - Colorectal cancer is the second most common cause of cancer deaths in the United States. Vitamin E (VE) and other antioxidants may help prevent colon cancer by decreasing the formation of mutagens arising from the free radical oxidation of fecal lipids or by "non-antioxidant" mechanisms. VE is not a single molecule, but refers to at least eight different molecules, that is, four tocopherols and four tocotrienols. METHODS: Both animal models and human colon cancer cell lines were used to evaluate the chemopreventive potential of different forms of VE. Rats were fed diets deficient in tocopherols or supplemented with either alpha tocopherol or gamma-tocopherol. Half the rats in each of these groups received normal levels of dietary Fe and the other half Fe at eight times the normal level. In our cell experiments, we looked at the role of gamma-tocopherol in upregulating peroxisome proliferator-activated receptor-gamma (PPAR-gamma) in the SW 480 human cell line. RESULTS: Rats fed the diets supplemented with alpha tocopherol had higher levels of VE in feces, colonocytes, plasma, and liver than did rats fed diets supplemented with gamma-tocopherol. Dietary Fe levels did not influence tocopherol levels in plasma, liver, or feces. For colonocytes, high dietary Fe decreased tocopherol levels. Rats fed the gamma-tocopherol supplemented diets had lower levels of fecal lipid hydroperoxides than rats fed the alpha-tocopherol-supplemented diets. Ras-p21 levels were significantly lower in rats fed the gamma-tocopherol-supplemented diets compared with rats fed the alpha-tocopherol-supplemented diets. High levels of dietary Fe were found to promote oxidative stress in feces and colonocytes. Our data with the SW480 cells suggest that both alpha- and gamma-tocopherol upregulate PPAR-gamma mRNA and protein expression. gamma-tocopherol was, however, found to be a better enhancer of PPAR-gamma expression than alpha-tocopherol at the concentrations tested. PMID- 15753150 TI - Vitamin E in preeclampsia. AB - Preeclampsia is the disorder of pregnancy with the highest rate of both maternal and neonatal morbidity and mortality. The maternal syndrome is characterized by oxidative stress and activation of the vascular endothelium that may originate from placental release of lipid peroxidation products, cytokines, and microparticles leading to an acute inflammatory response. The current understanding of the etiology has allowed the improvement of predictive tests, tests that could make intervention possible from early pregnancy onwards. Although the large secondary intervention antioxidant trials in cardiovascular diseases did not show any beneficial effect of vitamin E and vitamin C, either alone or in combination, knowledge of the nature of the pathogenesis of preeclampsia offers hope for the beneficial use of antioxidants in the prevention of the disorder. Not only has our previous small trial shown that antioxidant prophylactics in high-risk women lowered the prevalence of preeclampsia, but also new evidence has demonstrated multiple other actions of alpha-tocopherol (such as anti-inflammation and inhibition of NAD(P)H oxidase activation) besides its antioxidant properties that could be advantageous in the prevention of the disorder. Several larger trials are under way to investigate the precise role that vitamins C and E can play in the prevention of preeclampsia. PMID- 15753151 TI - Vitamin E in neurodegenerative disorders: Alzheimer's disease. AB - Oxidative stress is important in the pathogenesis of Alzheimer's disease (AD). The brain contains high levels of oxidizable lipids that must be protected by antioxidants. Low concentrations of vitamin E, quantitatively the major lipophilic antioxidant in the brain, are frequently observed in cerebrospinal fluid (CSF) of AD patients, suggesting that supplementation with vitamin E might delay the development of AD. In a placebo-controlled trial, vitamin E (2000 IU/day, 2 years) slowed (-53%) functional deterioration in patients with moderate AD (Sano et al., N. Engl. J. Med. 336: 1216-1222, 1997). Recently, use of vitamin E and vitamin C supplements in combination was found to be associated with reduced prevalence (-78%) and incidence (-64%) of AD in elderly population (Zandi et al., Arch. Neurol. 61: 82-88, 2004). These results are consistent with the ability of the supplementation with vitamin E (400 IU/day, 1 month) to increase its levels in CSF (123%) and plasma (145%) of AD patients and, in combination with vitamin C (1000 g/day), to decrease the susceptibility of CSF lipoproteins (up to -32%) to in vitro oxidation (Kontush et al., Free Radic. Biol. Med. 31: 345-354, 2001). In addition, vitamin E reduced lipid peroxidation and amyloid deposition in a transgenic mice model of AD (Sung et al., FASEB J. 18: 323-325, 2004). Computer modeling of the influence of vitamin E on lipoprotein oxidation reveals that the vitamin develops antioxidative activity in CSF lipoproteins in the presence of physiologically relevant, low amounts of oxidants. By contrast, under similar conditions, vitamin E behaves as a pro-oxidant in plasma lipoproteins, consistent with the model of tocopherol-mediated peroxidation (Stocker, Curr. Opin. Lipidol. 5: 422-433, 1994). This distinction is related to major differences in the levels of vitamin E (50 nM vs. 30 microM) and oxidizable lipids (4 microM vs. 2.5 mM) between CSF and plasma, which result in major differences in oxidative conditions (per unit of vitamin E) between CSF and plasma in the presence of similar amounts of oxidants. Altogether, these data suggest that vitamin E may be effective against in vivo oxidation of CSF lipoproteins and brain lipids, and offer new perspectives in the treatment of AD and other neurodegenerative disorders. PMID- 15753152 TI - Vitamin E in neural and visual function. AB - A rat model of vitamin E (alpha-tocopherol) deficiency with similar "clinical," electrophysiological, and neuropathological abnormalities to those seen in man was used to investigate the effects of various amounts and forms of alpha tocopheryl acetate (alphaTA) on neural and visual function. Electrophysiological techniques provide an objective, non-invasive measure of neural and visual function. These techniques were used in the animal model to determine the minimum dietary requirement of vitamin E necessary to prevent neural and visual abnormalities. They were also used to compare the biological activities of the natural (RRR-) and synthetic (all-rac-) forms of alpha-tocopherol in neural tissues. The results were as follows: (1) Significant differences in neural and visual function were observed between deficient and control rats after approximately 8 months. (2) An intake of 1.0 mg/kg all-rac- or 0.75 mg/kg RRR alphaTA was observed to marginally protect nerves from vitamin E deficiency. (3) The biological activity of all-rac-alpha-tocopherol in neural tissues was approximately 75% of RRR-alpha-tocopherol. (4) The concentration of free malondialdehyde (an indicator of lipid peroxidation) was significantly increased in tissues from the deficient compared to the control animals. These results are consistent with a deficiency of alpha-tocopherol causing increased lipid peroxidation leading to abnormal neural electrophysiology. They could also be explained by more specific but as yet undefined function(s) of alpha-tocopherol in neural tissues. PMID- 15753153 TI - Vitamin E modulation of cardiovascular disease. AB - Endothelium in the vascular system is an important modulator of vasomotor tone and coagulation, and it plays a crucial role in the inhibition of adhesion and activation of platelets and leukocytes. Evidence indicates that dietary antioxidants may modulate these endothelium-dependent vascular functions through several mechanisms and may contribute to the prevention of vascular diseases such as atherosclerosis. Several cell cultures as well as animal and human clinical and observational studies have tested the efficacy of vitamin E on vascular function and the prevention of atherosclerosis. Our cell culture studies have indicated that vitamin E (alpha-tocopherol) inhibits the activation of endothelial cells stimulated by high levels of low-density lipoprotein cholesterol and pro-inflammatory cytokines. This inhibition is associated with the suppression of chemokines, the expression of cell surface adhesion molecules, and the adhesion of leukocytes to endothelial cells, all of which contribute to the development of lesions in the arterial wall. The molecular mechanisms by which alpha-tocopherol and other tocopherols modulate endothelial cells and smooth muscle functions have been delineated. We, and others, have also demonstrated a positive effect of dietary vitamin E on endothelium and vascular function in animal models of atherosclerosis. Several human clinical trials have also shown an improvement in the surrogate markers of atherosclerosis and vascular function by vitamin E supplementation. However, these findings have been contradicted by several vitamin E supplementation trials for the prevention of secondary cardiovascular events showing null effect. Intervention at a relatively late stage of disease and the single use of vitamin E rather than in combination with other antioxidants might have contributed to these contradictory findings. Evidence from cell cultures, as well as animal and human clinical and observational studies, strongly supports the contribution of dietary vitamin E to the maintenance of vascular function and health, in particular when it is used in combination with other dietary antioxidants, which are found in fruits, vegetables, and nuts. PMID- 15753154 TI - Vitamin E and cardiovascular disease: observational studies. AB - Basic research suggests that oxidative stress may play an important role in many chronic diseases and provides plausible mechanisms by which natural antioxidants such as vitamin E may delay or prevent steps in atherogenesis. Dietary research has shown that those who consume higher amounts of fruits and vegetables have lower rates of heart disease and stroke, raising the possibility that antioxidants are protective. Results from large-scale human observational studies suggest that antioxidant consumption reduces the risk of developing cardiovascular disease (CVD). Both case-control and prospective cohort studies have carefully explored the relationship between vitamin E intake and plasma and tissue vitamin E levels and the risk of CVD. In many, but not all, of these studies vitamin E intake over an extended period was associated with decreased risk of cardiovascular events. Results from studies of blood levels are more limited and less consistent. This presentation summarizes data from the major observational studies. Overall, they support the possibility that vitamin E intake either from food or supplements may reduce risk of CVD; however, these studies have important limitations. For example, uncontrolled confounding can be similar in magnitude to the observed health effects, and antioxidant consumption may be merely a marker for a different cardioprotective factor (such as exercise or diet) that is responsible for these effects. In the search for small to moderate effects, randomized trials may be helpful, although to date, data from large-scale trials have been inconsistent. Several large-scale trials currently under way will help identify the potential benefits of vitamin E in the primary prevention of CVD and other chronic illness. Some are designed to test vitamin E alone as well as in combination with other antioxidant supplements because it is possible that antioxidants may be most effective if taken in particular combinations. Currently, the American Heart Association maintains that there are insufficient efficacy data from completed randomized trials to justify population wide recommendations for use of vitamin E supplements in disease prevention. PMID- 15753155 TI - Vitamin E for the treatment of cardiovascular disease: is there a future? AB - Oxidative stress seems to play a key role in the pathogenesis of atherosclerosis. Agents that protect low-density lipoprotein from oxidation have been shown in a range of in vitro and animal models to reduce the development and progression of atherosclerosis. These agents include antioxidant micronutrients such as vitamin E. They have gained wide interest because of the potential for prevention of atherosclerotic vascular disease in humans. In the last decade, many trials with antioxidants have been carried out in patients with cardiovascular disease, but the results are equivocal. The reason for the disappointing findings is unclear, but one possible explanation is the lack of identification criteria of patients who are potential candidates for antioxidant treatment. This review analyses the data reported so far to determine whether they clearly support the premise that patients at risk of cardiovascular disease may be candidates for antioxidant treatment. PMID- 15753156 TI - Future directions in preclinical vitamin E research: panel discussion A. PMID- 15753157 TI - Future directions in clinical vitamin E research: panel discussion B. PMID- 15753158 TI - Fluorescent tocopherols as probes of inter-vesicular transfer catalyzed by the alpha-tocopherol transfer protein. AB - Novel fluorescent analogues of alpha-tocopherol have been prepared that incorporate the useful fluorophores nitrobenoxadiazyl (NBD) and anthroyloxy (AO). Both fluorescent tocopherol analogues bind specifically to recombinant human tocopherol transfer protein (hTTP). The NBD-alpha-tocopherol is particularly useful for protein-binding assays, whereas the AO-alpha-tocopherol was designed to be one of a pair of chromophores for a fluorescence resonance energy transfer (FRET) assay of intervesicular tocopherol transfer. It is now possible to follow AO-alpha-tocopherol transfer from donor lipid vesicles composed of predominantly phosphatidylcholine (PC) to acceptor lipid vesicles containing PC and a quenching lipid NBD-PE (2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[7-nitro-2-1,3 benzoxadiazol-4-yl]). The presence of hTTP substantially increases the rate of AO alpha-tocopherol transfer over the uncatalyzed spontaneous rate. PMID- 15753159 TI - Gene-nutrient interactions exemplified by the alpha-tocopherol content of tissues from alpha-tocopherol transfer protein-null mice fed different dietary vitamin E concentrations. PMID- 15753160 TI - Intracellular localization of alpha-tocopherol transfer protein and alpha tocopherol. AB - The mechanism of action of tocopherol transfer protein (TTP) and its role in the intracellular processing of vitamin E were investigated using confocal fluorescence microscopy. The results from this work suggest that TTP functions by transporting vitamin E from endocytic organelles to other locations in the cell. PMID- 15753161 TI - Structure-function relationship in the tocopherol transfer protein. AB - The role of specific amino acid residues in mediating the biochemical functions of tocopherol transfer protein (TTP) was investigated using site-directed mutagenesis and functional assays. These findings further current understanding of TTP mechanism of action and its role in human health. PMID- 15753162 TI - Alpha-tocopherol affects androgen metabolism in male rat. AB - The Alpha-Tocopherol Beta-Carotene Cancer Prevention Study has provided the first evidence implicating vitamin E in hormone synthesis. The effect of vitamin E on stereoidogenesis in testes and adrenal glands was assessed in growing rats using Affymetrix gene-chip technology. Dietary supplementation of rats with vitamin E (60 mg/kg feed) for a period of 429 days caused a significant repression of genes encoding for proteins centrally involved in the uptake (low-density lipoprotein receptor) and de novo synthesis (for example, 7-dehydrocholesterol reductase, 3 hydroxy-3-methylglutaryl coenzyme A synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase, isopentenyl-diphosphate delta-isomerase, and farnesyl pyrophosphate synthetase) of cholesterol, the precursor of all steroid hormones. The present investigation indicates that dietary vitamin E may induce changes in stereoidogenesis by affecting cholesterol homeostasis. PMID- 15753163 TI - The transcriptional signature of vitamin E. AB - To investigate the ability of RRR-alpha-tocopherol to regulate gene expression, we interrogated RNA samples from treated cultured cells with Affymetrix oligonucleutodie arrays. We find that vitamin E potently regulates the expression of ca. 230 genes, functioning in metabolism, cell-cycle progression, and transcriptional regulation. PMID- 15753164 TI - Alpha- and gamma-tocopherol plasma and urinary biokinetics following alpha tocopherol supplementation. PMID- 15753165 TI - Oxidized vitamin E and ubiquinone: competition for binding sites of the mitochondrial cytochrome bc1 complex? PMID- 15753166 TI - Antioxidant properties of chromanols derived from vitamin E and ubiquinone. PMID- 15753167 TI - Vitamin E in uremia and dialysis patients. AB - Vitamin E therapy (based either on oral supplements or new dialysis methods such as vitamin E-coated hemodialysers) has been suggested to yield a better clinical outcome in hemodialysis (HD) patients than in other populations of patients. Among other factors, the presence of a modified vitamin E status might help to explain this apparently paradoxical response to vitamin E. In this study we investigated 104 regular HD patients. The results indicate that, besides having a low dietary intake, these subjects show some abnormalities in the levels and metabolism of vitamin E, such as a disproportion between plasma tocopherols and lipids, low levels of gamma-T, and CEHC accumulation. Although further studies are needed to confirm the clinical relevance of vitamin E therapy in HD, these findings might lead to recommending a higher vitamin E intake in these patients. PMID- 15753168 TI - Oxidative stress and changes in alpha- and gamma-tocopherol levels during coronary artery bypass grafting. AB - We studied whether cardiopulmonary bypass (CPB) has any immediate impact on the initiation of antioxidative defenses in the body by measuring F(2)-isoprostanes and alpha- and gamma-tocopherol, respectively. 8-iso-PGF(2alpha) levels increased significantly within 3 minutes and until the end of CPB. alpha-Tocopherol levels increased gradually at 20 min during CPB and continued until 6 hours after CPB. gamma-Tocopherol levels followed a similar fashion at the end of CPB. 8-iso PGF(2alpha) and tocopherol levels kept at basal level 12 and 24 hours post CPB. These findings suggest that an increased free radical-induced oxidative stress together with a gradual appearance of antioxidative defense system during and after CPB. PMID- 15753169 TI - Cigarette smoking increases human vitamin E requirements as estimated by plasma deuterium-labeled CEHC. AB - Cigarette smoking (CS) is a well-described oxidant burden in humans. We hypothesized that CS would accelerate alpha-tocopherol (alpha-T) utilization leaving less for metabolite (CEHC) production. After labeled alpha-T consumption (75 mg each of d(3)-RRR-alpha-TAc and d(6)-all-rac-alpha-TAc) by smokers and nonsmokers (n = 10/group), CS increased alpha-T disappearance and decreased plasma and urinary CEHCs. Plasma d(3)/d(6)-alpha-T ratios were approximately 1.4 during supplementation and approximately 2 from days 5 to 17. d(3)/d(6)-alpha CEHC ratios were on average 0.29 +/- 0.05, confirming that all-rac-alpha tocopherol is metabolized more efficiently. CEHC may be a good marker of vitamin E status, and smokers may have an increased vitamin E requirement. PMID- 15753170 TI - Effects of vitamin E depletion/repletion on biomarkers of oxidative stress in healthy aging. AB - The effects on ex vivo LDL resistance to oxidation and biomarkers of in vivo oxidative stress in response to 3-month dietary vitamin E restriction to 25% of recommended intake and 2-month unrestricted dietary intake and supplementation with 800 IU/d were studied in 100 healthy, nonsmoking 20-75-year-old volunteers. Significant changes in vitamin E status were associated with decreases and increases, respectively, in LDL resistance to oxidation in the depletion and supplementation period and with decreases in lipid peroxidation and oxidative DNA modification in the supplementation period. Healthy aging was not associated with enhanced susceptibility to oxidation in the depletion period. PMID- 15753171 TI - Consumption of sesame oil muffins decreases the urinary excretion of gamma tocopherol metabolites in humans. AB - Sesame seed and oil consumption previously increased human plasma gamma tocopherol (gamma-T) concentrations. This was attributed to the sesame lignans sesamin and sesamolin. Here, we studied the inhibition of vitamin E metabolism by a single dose of sesame oil lignans coingested with deuterated alpha- and gamma tocopherols in human volunteers. The urinary excretion of gamma-T metabolites was significantly lower in sesame oil treated than in control subjects. Concentrations of tocopherols in blood were not affected by the treatment. In conclusion, a single dose of sesame oil, containing 136 mg sesame lignans (sesamin and sesamolin), reduces the urinary excretion of co-administered gamma-T in humans. PMID- 15753172 TI - Characterization of cellular uptake and distribution of vitamin E. AB - We previously reported that tocotrienols acted as more potent inhibitors against selenium deficiency-induced cell death than the corresponding tocopherol isoforms (J. Biol. Chem. 2003;278:39428-39434). In the present study, we first compared the differences in the cellular uptake between alpha-tocopherol (alpha-Toc) and alpha-tocotrienol (alpha-Toc-3). The initial rate of cellular uptake of alpha-Toc 3 was 70-fold higher than that of alpha-Toc. Subcellular fractionation analysis of alpha-Toc-3 and alpha-Toc-fortified cells showed similar cellular distribution of these antioxidants, which was directly proportional to the lipid distribution. The cells containing similar amounts of alpha-Toc-3 and alpha-Toc showed similar resistance against the oxidative stress caused by peroxides. These results suggest that the apparent higher cytoprotective effect of alpha-Toc-3 than alpha Toc is primarily ascribed to its higher cellular uptake. PMID- 15753173 TI - Vitamin E exhibits concentration- and vitamer-dependent impairment of microsomal enzyme activities. PMID- 15753174 TI - The decrease in gamma-tocopherol in plasma and lipoprotein fractions levels off within two days of vitamin E supplementation. AB - The effects of vitamin E supplementation on alpha- and gamma-tocopherol concentrations were studied in plasma and lipoprotein fractions of five healthy volunteers taking 1000 IU/day of RRR alpha-tocopherol for 4 days. Although plasma alpha-tocopherol increased, gamma-tocopherol decreased. Compared with baseline, gamma-/alpha-tocopherol ratios decreased from 48 h onward (P < 0.001). They all leveled off within 48 h. From 12 h onward, gamma-/alpha-tocopherol ratios were higher in VLDL and IDL than in LDL and HDL, indicating that gamma-tocopherol is better maintained in triglyceride-rich lipoprotein fractions. These data suggest that vitamin E supplementation exceeding 2 days does not further decrease gamma tocopherol concentrations. PMID- 15753175 TI - Does aging affect the response of vitamin E status to vitamin E depletion and supplementation? AB - A vitamin E depletion/supplementation study was conducted in 100 healthy 20-75 year-old volunteers. The responses of vitamin E status to 3-week dietary vitamin E restriction to approximately 25% of recommended intake and 2-month unrestricted dietary intake plus 800 IU/d of RRR-alpha-tocopherol were studied as a function of age. Plasma alpha-tocopherol concentrations were closely related to cholesterol concentrations, which increased with age (P < 0.001). Upon dietary restriction, plasma alpha-tocopherol concentrations decreased significantly (P < 0.001) but independently of age. Plasma alpha-tocopherol responses to supplementation increased significantly with age, but this effect disappeared after standardization for cholesterol. gamma-Tocopherol concentrations decreased to less than 30% of baseline. PMID- 15753177 TI - Current status of vitamin E nutriture. AB - Vitamin E intake status requires reassessment because the recommended levels have been increased and take into account only the alpha-tocopherol form of vitamin E. A database of alpha-tocopherol values for more than 7,000 foods was developed and applied to dietary data from the National Health and Nutrition Examination Survey 1999-2000. Usual intake distributions were determined and evaluated for adequacy. Ninety percent or more of the adults studied had their usual intakes below the current Estimated Average Requirement. Several observations-the prevalence of inadequate intakes of vitamin E, absence of signs of deficiency in the U.S. population, and increasing evidence that vitamin E helps reduce chronic disease risk-point to a need for further research. PMID- 15753176 TI - The maximal amount of alpha-tocopherol intake from foods alone in U.S. adults (1994-1996 CSFII): an analysis by linear programming. PMID- 15753178 TI - Gamma-tocotrienol metabolism and antiproliferative effect in prostate cancer cells. AB - In this study, we evaluated the antiproliferative effect of tocotrienols (T3) and the presence of a specific vitamin E metabolism in PC3 and LNCaP prostate cancer cells. These cell lines are able to transform tocopherols (T) and T3 in the corresponding carboxyethyl-hydroxychromans metabolites (CEHCs). The extent of this metabolism and the inhibitory effect on cell growth followed the order of magnitude alpha-T/=2 SDs higher than the normal resting heart rate for their age and sex) at ED triage were more likely to meet criteria for partial or full PTSD at follow-up, even after adjusting for age, sex, and injury (adjusted odds ratio, 2.4 [95% confidence interval, 1.1-5.4]). CONCLUSION: These results suggest an association between early physiological arousal and the development or persistence of PTSD symptoms in injured children and point to the importance of better understanding the interplay between physiological and psychological functioning after a traumatic stressor. PMID- 15753249 TI - Development of immunoglobulin M capture enzyme-linked immunosorbent assay to differentiate human flavivirus infections occurring in Australia. AB - We report the development of a flavivirus immunoglobulin M (IgM) capture enzyme linked immunosorbent assay (MAC-ELISA) which improves the determination of an infecting flavivirus serotype over that by current serological methods. A panel of 165 IgM-positive sera from flavivirus patients with specific diagnostic results was tested by the flavivirus MAC-ELISA using a panel of 10 antigens. For 134 of these sera (81.2%), the highest reactivity was demonstrated against the infecting virus, which was consistent with the original diagnostic result. Specific antibody reactions inconsistent with the original diagnosis were found for six sera (3.6%). In our experience, the flavivirus-serotyping ELISA provides a rapid and accurate alternative to other serological tests, such as hemagglutination inhibition, for the specific diagnosis of flavivirus infections. PMID- 15753248 TI - Evaluation of three immunoassay kits for rapid detection of influenza virus A and B. AB - Influenza causes high morbidity and mortality in very young and elderly individuals, which can be controlled with antivirals and/or vaccines. The success of therapeutic measures is predicated on the rapid and precise diagnosis of the infection. We compared three rapid influenza immunoassay (RIIA) kits for the diagnosis of influenza virus A and B using 178 respiratory specimens submitted for routine testing. BD Directigen Flu A+B (Directigen), Directigen EZ Flu A+B (EZ), and NOW Flu A NOW Flu B (NOW; Binax) tests had comparable combined influenza virus A and B specificities, varying from 94 to 98%. In contrast, the sensitivity of EZ was significantly lower (39%) than that of NOW (76%) and marginally lower than that of Directigen (56%). The differences in sensitivity were most evident in patients who were >9 years old (Directigen, 53%; EZ, 32%; and NOW, 69%). Among specimens, bronchoalveolar lavage fluids yielded the most discrepant results, with sensitivities varying from 0 (EZ) to 100% (NOW), followed by nasopharyngeal swabs (sensitivities of 27 to 100%) and nasal washes (50 to 81%). The Directigen kit format allowed for faster completion but more cumbersome performance and more difficult interpretation compared with the other two kits. Overall, NOW provided the most accurate diagnoses and had user-friendly technical characteristics. However, the low overall sensitivity of the RIIAs indicates that these can be used as screening tools only. PMID- 15753250 TI - Placental malaria diminishes development of antibody responses to Plasmodium falciparum epitopes in infants residing in an area of western Kenya where P. falciparum is endemic. AB - To determine the effect of placental malaria (PM) infection on the development of antibody responses to malaria in infants, we measured immunoglobulin G levels to seven different Plasmodium falciparum epitopes by using plasma samples collected at monthly intervals from infants born to mothers with and without PM. Overall, PM was associated with diminished antibody levels to all of the epitopes tested, especially with infants aged >or=4 to 12 months, and the difference was statistically significant for four of the seven epitopes (P<0.0035). These findings suggest that PM can negatively influence the development of immune responses to malaria in infants. PMID- 15753251 TI - Application of monoclonal antibodies in functional and comparative investigations of heavy-chain immunoglobulins in new world camelids. AB - Of the three immunoglobulin G (IgG) isotypes described to occur in camelids, IgG2 and IgG3 are distinct in that they do not incorporate light chains. These heavy chain antibodies (HCAbs) constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. We have produced isotype-specific mouse monoclonal antibodies (MAbs) in order to investigate the roles of HCAbs in camelid immunity. Seventeen stable hybridomas were cloned, and three MAbs that were specific for epitopes on the gamma chains of llama IgG1, IgG2, or IgG3 were characterized in detail. Affinity chromatography revealed that each MAb bound its isotype in solution in llama serum. The antibodies bound to the corresponding alpaca IgGs, to guanaco IgG1 and IgG2, and to camel IgG1. Interestingly, anti IgG2 MAbs bound three heavy-chain species in llama serum, confirming the presence of three IgG2 subisotypes. Two IgG2 subisotypes were detected in alpaca and guanaco sera. The MAbs detected llama serum IgGs when they were bound to antigen in enzyme-linked immunosorbent assays and were used to discern among isotypes induced during infection with a parasitic nematode. Diseased animals, infected with Parelaphostrongylus tenuis, did not produce antigen-specific HCAbs; rather, they produced the conventional isotype, IgG1, exclusively. Our data document the utility of these MAbs in functional and physiologic investigations of the immune systems of New World camelids. PMID- 15753252 TI - Comparison of antibody repertoires against Staphylococcus aureus in healthy individuals and in acutely infected patients. AB - The management of staphylococcal diseases is increasingly difficult with present medical approaches. Preventive and therapeutic vaccination is considered to be a promising alternative; however, little is known about immune correlates of protection and disease susceptibility. To better understand the immune recognition of Staphylococcus aureus by the human host, we studied the antistaphylococcal humoral responses in healthy people in comparison to those of patients with invasive diseases. In a series of enzyme-linked immunosorbent assay analyses performed using 19 recombinant staphylococcal cell surface and secreted proteins, we measured a wide range of antibody levels, finding a pronounced heterogeneity among individuals in both donor groups. The analysis revealed marked differences in the antibody repertoires of healthy individuals with or without S. aureus carriage, as well as in those of patients in the acute phase of infection. Most importantly, we identified antigenic proteins for which specific antibodies were missing or underrepresented in infected patients. In contrast to the well-described transient nature of disease-induced antistaphylococcal immune response, it was demonstrated that high-titer antistaphylococcal antibodies are stable for years in healthy individuals. In addition, we provide evidence obtained on the basis of opsonophagocytic and neutralizing activity in vitro assays that circulating antistaphylococcal serum antibodies in healthy donors are functional. In light of these data we suggest that proper serological analysis comparing the preexisting antibody repertoires of hospitalized patients with different outcomes for nosocomial staphylococcal infections could be extremely useful for the evaluation of candidate vaccine antigens in addition to protection data generated with animal models. PMID- 15753253 TI - Specific inhibitory action of anisodamine against a staphylococcal superantigenic toxin, toxic shock syndrome toxin 1 (TSST-1), leading to down-regulation of cytokine production and blocking of TSST-1 toxicity in mice. AB - Toxic shock syndrome toxin 1 (TSST-1), produced by Staphylococcus aureus (including methicillin-resistant S. aureus), is a superantigenic toxin responsible for toxic shock syndrome as well as neonatal TSS-like exanthematous disease. TSST-1 exhibits its deleterious effects by leading to the abnormal proliferation of, e.g., Vbeta2+ T cells and overproduction of proinflammatory cytokines. In the present study we examined the inhibitory effect of a Chinese herbal extract, anisodamine, on TSST-1 using human peripheral blood mononuclear cells (PBMCs). Anisodamine inhibited the production of proinflammatory cytokines better than interleukin-10 (an anti-inflammatory cytokine). The inhibitory effect of anisodamine was greater than that of any tropane alkaloid examined. Anisodamine acted directly on both monocytes and T cells in human PBMCs, and the effect was confirmed at the transcriptional level. Inhibition of NF-kappaB activation was also demonstrated. In contrast, no significant inhibition of Vbeta2+ T-cell proliferation was observed. In mice injected with TSST-1, anisodamine treatment significantly decreased serum proinflammatory cytokine levels and prevented TSST-1-induced death. These results suggest that anisodamine specifically acts against the production of cytokines (inflammatory cytokines in particular) and not against Vbeta2+ T-cell proliferation and that anisodamine may have a beneficial effect on TSST-1-associated disease. PMID- 15753255 TI - Development of a nanoparticle-labeled microfluidic immunoassay for detection of pathogenic microorganisms. AB - The light-scattering properties of submicroscopic metal particles ranging from 40 to 120 nm in diameter have recently been investigated. These particles scatter incident white light to generate monochromatic light, which can be seen either by the naked eye or by dark-field microscopy. The nanoparticles are well suited for detection in microchannel-based immunoassays. The goal of the present study was to detect Helicobacter pylori- and Escherichia coli O157:H7-specific antigens with biotinylated polyclonal antibodies. Gold particles (diameter, 80 nm) functionalized with a secondary antibiotin antibody were then used as the readout. A dark-field stereomicroscope was used for particle visualization in poly(dimethylsiloxane) microchannels. A colorimetric quantification scheme was developed for the detection of the visual color changes resulting from immune reactions in the microchannels. The microchannel immunoassays reliably detected H. pylori and E. coli O157:H7 antigens in quantities on the order of 10 ng, which provides a sensitivity of detection comparable to those of conventional dot blot assays. In addition, the nanoparticles within the microchannels can be stored for at least 8 months without a loss of signal intensity. This strategy provides a means for the detection of nanoparticles in microchannels without the use of sophisticated equipment. In addition, the approach has the potential for use for further miniaturization of immunoassays and can be used for long-term archiving of immunoassays. PMID- 15753254 TI - Acellular pertussis vaccine protects against exacerbation of allergic asthma due to Bordetella pertussis in a murine model. AB - The prevalence of asthma and allergic disease has increased in many countries, and there has been speculation that immunization promotes allergic sensitization. Bordetella pertussis infection exacerbates allergic asthmatic responses. We investigated whether acellular pertussis vaccine (Pa) enhanced or prevented B. pertussis-induced exacerbation of allergic asthma. Groups of mice were immunized with Pa, infected with B. pertussis, and/or sensitized to ovalbumin. Immunological, pathological, and physiological changes were measured to assess the impact of immunization on immune deviation and airway function. We demonstrate that immunization did not enhance ovalbumin-specific serum immunoglobulin E production. Histopathological examination revealed that immunization reduced the severity of airway pathology associated with sensitization in the context of infection and decreased bronchial hyperreactivity upon methacholine exposure of infected and sensitized mice. These data demonstrate unequivocally the benefit of Pa immunization to health and justify selection of Pa in mass vaccination protocols. In the absence of infection, the Pa used in this study enhanced the interleukin-10 (IL-10) and IL-13 responses and influenced airway hyperresponsiveness to sensitizing antigen; however, these data do not suggest that Pa contributes to childhood asthma overall. On the contrary, wild-type virulent B. pertussis is still circulating in most countries, and our data suggest that the major influence of Pa is to protect against the powerful exacerbation of asthma-like pathology induced by B. pertussis. PMID- 15753256 TI - Characterization of virus-responsive plasmacytoid dendritic cells in the rhesus macaque. AB - Plasmacytoid dendritic cells (PDC) are potent producers of alpha interferon (IFN alpha) in response to enveloped viruses and provide a critical link between the innate and adaptive immune responses. Although the loss of peripheral blood PDC function and numbers has been linked to human immunodeficiency virus (HIV) progression in humans, a suitable animal model is needed to study the effects of immunodeficiency virus infection on PDC function. The rhesus macaque SIV model closely mimics human HIV infection, and recent studies have identified macaque PDC, potentially making the macaque a good model to study PDC regulation. In this study, we demonstrate that peripheral blood PDC from healthy macaques are both phenotypically and functionally similar to human PDC and that reagents used for human studies can be used to study macaque PDC. Both human and macaque PBMC expressed IFN-alpha in response to herpes simplex virus (HSV), the prototypical activator of PDC, as measured by using an IFN bioassay and IFN-alpha-specific enzyme-linked immunospot assays. Similar to human PDC, macaque PDC were identified by using flow cytometry as CD123+ HLA-DR+ lineage- cells. In addition, like human PDC, macaque PDC expressed intracellular IFN-alpha, tumor necrosis factor alpha, macrophage inflammatory protein 1beta/CCL4, and IFN-inducible protein 10/CXCL10 upon stimulation with HSV, all as determined by intracellular flow cytometry. We found that IFN regulatory factor 7, which is required for the expression of IFN-alpha genes, was, similar to human PDC, expressed at high levels in macaque PDC compared to monocytes and CD8+ T cells. These findings establish the phenotypic and functional similarity of human and macaque PDC and confirm the utility of tools developed for studying human PDC in this animal model. PMID- 15753257 TI - Interleukin-18 primes the oxidative burst of neutrophils in response to formyl peptides: role of cytochrome b558 translocation and N-formyl peptide receptor endocytosis. AB - Using flow cytometry, we observed that interleukin-18 (IL-18) primed human neutrophils (PMNs) in whole blood to produce superoxide anion (O2 degrees-) in response to N-formyl peptide (fMLP) stimulation, whereas IL-18 alone had no significant effect. In contrast to tumor necrosis factor alpha (TNF-alpha), which is a cytokine known to strongly prime O2 degrees- production, IL-18 did not induce either p47phox phosphorylation or its translocation from the cytosol to the plasma membrane. However, IL-18 increased PMN degranulation, as shown by increased levels of cytochrome b558 and CD11b expression at the PMN surface. Moreover, addition of IL-18 to whole blood for 45 min reduced the ability of PMNs to bind to fMLP, suggesting endocytosis of fMLP receptors, as visualized by confocal microscopy. 2,3-Butanedione 2-monoxime, which inhibits endosomal recycling of plasma membrane components back to the cell surface, concomitantly accentuated the diminution of fMLP binding at the PMN surface and increased IL-18 priming of O2 degrees- production by PMNs in response to fMLP. This suggests that fMLP receptor endocytosis could account, at least in part, for the priming of O2 degrees- production. In addition, genistein, a tyrosine kinase inhibitor, and SB203580, a p38 mitogen-activated protein kinase (p38MAPK) inhibitor, completely reversed the decreased level of fMLP binding and increased the level of CD11b expression after IL-18 treatment. Flow cytometric analysis of intact PMNs in whole blood showed that IL-18 increased p38MAPK phosphorylation and tyrosine phosphorylation. In particular, IL-18 induced phosphorylation of focal adhesion kinase (p125FAK), which has been implicated in cytoskeleton reorganization. Taken together, our findings suggest several mechanisms that are likely to regulate cytokine-induced priming of the oxidative burst in PMNs in their blood environment. PMID- 15753258 TI - Kinetics of local and systemic immune responses after vaginal immunization with recombinant cholera toxin B subunit in humans. AB - Vaginal vaccination seems to be the best strategy for inducing specific immunoglobulin A (IgA) and IgG antibody responses in the female genital tract. The relative efficiencies of one, two, and three vaginal doses of recombinant cholera toxin B subunit (CTB) in generating mucosal and systemic immune responses in healthy women were evaluated, and the kinetics of the immune responses were monitored for responding volunteers for up to 12 months after the last vaccination. A single dose of CTB failed to generate CTB-specific IgA antibody responses in cervical secretions. Two vaccinations induced significant increases in IgA antitoxin titers in seven of nine volunteers, and four volunteers also developed IgG antitoxin responses. The magnitudes of the responses were 20-fold for IgA antitoxin and 7.1-fold for IgG antitoxin. A third vaccination did not significantly increase the antitoxin responses, although the frequency of IgG responses was slightly higher than that after the second vaccination. In serum, CTB-specific antibodies were observed already after a single vaccination. However, two vaccinations were required to induce marked IgA as well as IgG antitoxin titer increases in the majority of volunteers. The postvaccination levels of antitoxin antibodies in serum were comparable after two and three vaccinations. At 12 months after vaccination, significantly elevated IgA and IgG antitoxin levels in cervical secretions could still be detected in approximately half of the volunteers who had initially responded to the vaccine. Antitoxin titer increases in serum were found in most of the vaccinees at follow-up. PMID- 15753259 TI - Impaired accessory cell function in a human dendritic cell line after human immunodeficiency virus infection. AB - We generated human dendritic cell (DC) hybridoma cell lines by fusing HGPRT deficient promonocytic U937 cells with immature DCs obtained by culturing peripheral blood monocytes with interleukin-4 (IL-4; 1,000 U/ml) and granulocyte macrophage colony-stimulating factor (100 U/ml) for 7 days and mature DCs by treatment with tumor necrosis factor alpha (12.5 microg/ml) for 3 days. Only one fusion with immature DCs was successful and yielded four cell lines--HB-1, HB-2, HB-3, and HB-9--with an overall fusion efficiency of 0.0015%. The cell lines were stable in long-term culture, displayed morphological features typical of DCs, and expressed distinct class I and class II molecules not present on U937 (A*031012, B*51011, Cw*0701, DRB3*01011 52, and DR5*01011). A representative cell line, HB 2, that expressed DC markers including CD83, CD80 and CD86 could be induced to produce IL-12 through CD40 stimulation. After human immunodeficiency virus (HIV) infection, there was impairment of antigen-presenting cell (APC) function, which was manifested by an inability to stimulate allogeneic T-cell responses. There was no change in expression of major histocompatibility complex class I and class II antigens, CD83, CD40, CD4, CD11c, CD80, CD86, CD54, and CD58, or IL-12 production in the HIV-infected HB-2 cells. The HIV-infected HB-2 cells induced T cell apoptosis in the cocultures. T-cell proliferation could be partially restored by using ddI, indinivir, and blocking anti-gp120 and anti-IL-10 antibodies. Our data suggest that there are multiple mechanisms that DCs use to inhibit T-cell responses in HIV-infected patients. The HB-2 cell line could be a useful model system to study APC function in HIV-infected DCs. PMID- 15753260 TI - Glycolipids of Mycobacterium tuberculosis strain H37Rv are potential serological markers for diagnosis of active tuberculosis. AB - A simple and cost-effective diagnostic tool (TB Screen Test) for the screening of patients with pulmonary and extrapulmonary tuberculosis and for differentiation of those individuals from individuals without tuberculosis, other common infections, and healthy controls has been developed. The serological responses of purified mycobacterial glycolipid antigens were examined by a liposome agglutination assay. The assay was able to detect very low antiglycolipid antibody concentrations in the infected individuals. The sera from the tuberculosis patient group had significantly higher concentrations of antiglycolipid antibody than the sera from uninfected control subjects, with 94% sensitivity and 98.3% specificity. Glycolipids of Mycobacterium tuberculosis H37Rv antigens were isolated, purified, and characterized. After interchelation with liposome particles, these purified antigens specifically bound to the antiglycolipid antibodies present in the sera of patients with tuberculosis, resulting in the formation of a blue agglutination. This protocol clearly differentiates healthy controls and M. bovis BCG-vaccinated subjects from those with active tuberculosis. The resultant diagnostic tool, the TB Screen Test, is more economical and rapid (4 min) than other currently available products and can be used for the mass screening of a heavily afflicted population. PMID- 15753261 TI - Use of the COOH portion of the nucleocapsid protein in an antigen-capturing enzyme-linked immunosorbent assay for specific and sensitive detection of severe acute respiratory syndrome coronavirus. AB - Antibody detection with a recombinant COOH portion of the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid (N) protein, N13 (amino acids 221 to 422), was demonstrated to be more specific and sensitive than that with the full-length N protein, and an N13-based antigen-capturing enzyme-linked immunosorbent assay providing a convenient and specific test for serodiagnosis and epidemiological study of SARS was developed. PMID- 15753262 TI - General vs subspecialty pediatrics: factors leading to residents' career decisions over a 12-year period. AB - OBJECTIVES: To determine factors motivating residents' career choices and to examine changes in these priorities over the last 12 years. During the last decade, surveys of pediatric training programs have shown trends toward residents choosing careers in general pediatrics rather than in subspecialties. Most recently, there is evidence of a shift back toward subspecialty careers. DESIGN: We surveyed past and present residents at The Children's Hospital of Philadelphia, Philadelphia, Pa, (training completion dates, 1991-2002) via an anonymous written questionnaire. RESULTS: The sample comprised 238 residents (mean +/- SD age, 30 +/- 3 years; 59% female, 41% male; 47% subspecialists, 53% generalists). Among the group as a whole, subject matter, role models, lifestyle issues, and teaching were the most important determinants for career choices. Less important were national trends, job openings, and research. When subspecialists and generalists were compared, both groups found subject matter to be their highest priority. Among residents interested in subspecialties, teaching, research, and technical skills were significant (P<.001), compared with generalists, who considered lifestyle and personal/financial issues more important (P<.001). Lifestyle issues were also more important to female residents, those 30 years of age or younger, and those completing training recently (P<.05). CONCLUSIONS: Career decisions for pediatric residents today are motivated by complex factors. For those choosing generalist careers, lifestyle and personal/financial considerations predominate, while teaching, research, and technical skills are key factors for subspecialists. Over the last decade, lifestyle issues have become a more dominant factor, particularly for women entering the pediatric workforce. PMID- 15753263 TI - Prevention of the epidemic increase in child risk of overweight in low-income schools: the El Paso coordinated approach to child health. AB - OBJECTIVE: To assess the impact on children's health of translating an evidence based national intervention trial (Child and Adolescent Trial for Cardiovascular Health [CATCH]) to low-income elementary schools with primarily Hispanic students. DESIGN: An untreated, matched control group design with repeated dependent pretest and posttest samples was used. SETTING: Four El Paso CATCH and 4 control elementary schools in El Paso, Tex, along the US-Mexico border region. All had Title I status (most were low-income students). PARTICIPANTS: Participants were 896 third-grade children (473 control schools [224 girls and 249 boys] and 423 CATCH schools [199 girls and 224 boys]); 93% were Hispanic. INTERVENTION: Community-based implementation of the national CATCH program. MAIN OUTCOME MEASURES: Risk of overweight or overweight, body mass index, waist-to-hip ratio, yards run in 9 minutes, passing rates for Fitnessgram national mile standards, moderate to vigorous physical activity and vigorous physical activity in physical education class, and percentage of fat and sodium in school lunches. RESULTS: Girls in control schools had significant increases in percentage of risk of overweight or overweight from third (26%) to fifth (39%) grades, as did girls in CATCH schools (30%-32%); however, the rate of increase for girls in the CATCH schools was significantly lower (2%) compared with the rate for control girls (13%). A similar pattern was seen for boys, with a rate of increase for boys in CATCH schools of 1% (40%-41%), which was significantly less than the 9% increase (40% to 49%) for control boys. CONCLUSIONS: The translation of the national CATCH program to low-income schools with Hispanic students successfully slowed the epidemic increase in risk of overweight or overweight seen in control school children. An emphasis should be placed on community organizing and evaluation feedback when implementing evidence-based school health programs in low-income Hispanic communities. PMID- 15753264 TI - The impact of a school-based obesity prevention trial on disordered weight control behaviors in early adolescent girls. AB - OBJECTIVE: To assess the impact of an obesity prevention intervention on use of self-induced vomiting/laxatives (purging) and diet pills to control weight in girls in early adolescence. DESIGN: We matched and randomly assigned 10 middle schools to an intervention or a control condition in a randomized controlled trial. Longitudinal multivariable analyses using generalized estimating equations were conducted with data from 480 girls to examine the effects of the intervention on the risk of reporting a new case of purging or diet pill use to control weight at follow-up 21 months later, while controlling for ethnicity and school matched pairs. Girls who reported purging or using diet pills at baseline were excluded from analyses. SETTING: Middle schools. PARTICIPANTS: Four hundred eighty girls in early adolescence aged 10 to 14 years (mean age, 11.5 years). INTERVENTION: The Planet Health obesity prevention program was implemented during 2 school years and was designed to promote healthful nutrition and physical activity and to reduce television viewing. OUTCOME: Reduced risk of using self induced vomiting/laxatives or diet pills to control weight in the past 30 days. RESULTS: After the intervention, we found 14 (6.2%) of 226 girls in control schools and 7 (2.8%) of 254 girls in intervention schools reported purging or using diet pills to control their weight (P = .003). In a multivariable generalized estimating equation model, girls in intervention schools were less than half as likely to report purging or using diet pills at follow-up compared with girls in control schools (odds ratio, 0.41; 95% confidence interval, 0.22 0.75). CONCLUSION: These findings provide promising evidence that school-based interventions may effectively integrate prevention of both obesity and disordered weight-control behaviors. PMID- 15753265 TI - Medical and psychiatric comorbidity and health care use among children 6 to 17 years old. AB - BACKGROUND: The association of psychiatric disorders (PDs) with other PDs and medical disorders (MDs) has been insufficiently explored in children and adolescents. OBJECTIVES: To estimate medical and psychiatric comorbidity present in children with PDs and to determine the medical service usage of children with PDs. DESIGN: We use administrative health care data to describe the health care provided for study children. Psychiatric disorders were classified into the following 3 categories: psychosis, emotion, and behavior. We used logistic regression to assess medical comorbidity for each category. Psychiatric comorbidity was determined using chi(2) test analysis. Health care use was determined by comparing the frequency of visits for MDs and PDs between children with PDs and children without PDs. SETTING: We studied 406,640 children (50.6% male) between 6 and 17 years old, living in Alberta, Canada, during the fiscal year April 1, 1995, through March 31, 1996. RESULTS: A PD was diagnosed in 32,214 (60.3% male) children. Psychiatric comorbidity was present in 13.6% of the children; comorbidity existed in all 3 psychiatric groups and peaked in postpubertal children. More girls than boys had significant medical comorbidity. Significant odds ratios (ORs) for girls varied from 1.2 (behavior and sinusitis, bronchitis, and chronic disorders; psychosis, and menstrual problems) to 15.3 (behavior and developmental delay). Among boys, the highest OR was seen with the combination of behavior and developmental delay (OR, 8.3) and psychosis and poisoning (OR, 8.2). With ORs ranging from 4.6 to 15.3, developmental delay consistently had high ORs for both sexes and all 3 types of PDs. Poisoning also had high ORs (3.3-14.1) with all 3 PDs and both sexes. Among girls, disorders associated with pregnancy and the genitourinary system had modest associations (OR, 1.9-2.2, for behavior) to moderate (OR, 2.5-4.0, for emotion). Children with PDs had significantly greater medical service usage than did children without PDs. Girls had greater medical health care usage than boys. Psychiatric service usage was similar for both sexes. CONCLUSIONS: Medical and psychiatric comorbidity exist in children with PDs. Girls are more commonly affected. Health care usage is higher in children with PDs. PMID- 15753266 TI - Efficacy of noninvasive tests in the diagnosis of Helicobacter pylori infection in pediatric patients. AB - BACKGROUND: Helicobacter pylori infection is likely acquired in childhood. Helicobacter pylori is recognized as a cause of gastritis and peptic ulcer. OBJECTIVE: To investigate some noninvasive tests, particularly H pylori fecal antigen, for the diagnosis of H pylori infection in comparison with the gold standard invasive test, esophagogastroduodenoscopy with biopsy. METHODS: We studied 250 patients (102 male; age range, 3-18 years) who underwent esophagogastroduodenoscopy with biopsy (histologic examination and rapid urease test) for a suspicious upper gastrointestinal disease; in all of them, fecal H pylori antigen, serum H pylori immunoglobulin G, and cytotoxin-associated gene product A immunoglobulin G were measured. Sensitivity and specificity of noninvasive tests were compared with those of the gold-standard esophagogastroduodenoscopy with biopsy. RESULTS: Ninety-three patients (37%) had positive histopathologic (Giemsa staining) and rapid urease test results. The H pylori fecal antigen revealed a sensitivity of 97%, a specificity of 98%, a positive predictive value of 97%, and a negative predictive value of 98%; serum H pylori immunoglobulin G had a sensitivity of 86%, a specificity of 80%, a positive predictive value of 72%, and a negative predictive value of 90%; and serum cytotoxin-associated gene product A immunoglobulin G had a sensitivity of 83%, a specificity of 80%, a positive predictive value of 71%, and a negative predictive value of 89%. CONCLUSIONS: Our study demonstrates that among noninvasive and easily applicable tests, particularly in small children, H pylori fecal test is simple, suitable, and has high accuracy for the screening of H pylori-positive patients. PMID- 15753267 TI - Factors associated with fragmented sleep at night across early childhood. AB - OBJECTIVE: To identify the factors most strongly associated with sleeping less than 6 consecutive hours at night for children aged 5, 17, and 29 months. DESIGN, SETTING, AND PARTICIPANTS: A randomized survey design used a representative sample of infants born in 1997-1998 in the Canadian province of Quebec. Data were collected by questionnaires and interviews. Interviews were scheduled at home with the mothers. The number of consecutive hours slept at night by 1741 children aged 5, 17, and 29 months was assessed from parental reports. Factors associated with fragmented sleep were investigated for each age in a cross-sectional design. RESULTS: At 5 months of age, 23.5% of children did not sleep 6 consecutive hours. Of the children who did not sleep 6 consecutive hours at night at 5 months or 17 months of age, 32.9% were still not sleeping 6 consecutive hours at night at 29 months of age. The factor most strongly associated with not sleeping at least 6 consecutive hours per night at 5 months of age was feeding the child after an awakening. Parental presence until sleep onset was the factor most strongly associated with not sleeping at least 6 consecutive hours per night at 17 months and 29 months of age. CONCLUSIONS: Sleep consolidation evolves rapidly in early childhood. Parental behaviors at bedtime and in response to a nocturnal awakening are highly associated with the child's sleep consolidation. The effects are probably bidirectional and probably create a long-term problem. Early interventions could possibly break the cycle. PMID- 15753268 TI - Computer-based documentation: effect on parent and physician satisfaction during a pediatric health maintenance encounter. AB - OBJECTIVE: To investigate the impact of a computer-based documentation (CBD) tool on parent and physician satisfaction with a pediatric health maintenance encounter. DESIGN: The project used a preintervention and postintervention design. The preintervention group visits used paper-based forms for data entry, whereas the postintervention visits used CBD. At the conclusion of each encounter, both the physician and the parent completed a survey that assessed their perceptions of the encounter's quality. SETTING: Urban hospital-based pediatric teaching clinic. PARTICIPANTS: Parents and physicians of children 18 months and younger. MAIN OUTCOME MEASURES: Parent and physician satisfaction with 7 components of a health maintenance encounter (interim history, social history, anticipatory guidance, developmental assessment, physical examination, assessment, and plan). RESULTS: There was no change in overall parent or physician satisfaction in the areas of communication or physician helpfulness. Physicians using CBD were less likely to agree that they provided clear explanations but were also less likely to agree that they "acted bossy during the visit." There was no correlation between physician and parent satisfaction or between physician satisfaction and pattern of CBD use. However, there was a strong correlation between physicians' satisfaction and the extent to which they found CBD helpful (Spearman rho = 0.29, P<.001). CONCLUSIONS: The introduction of CBD into the health maintenance encounter did not affect measured aspects of parent or physician satisfaction; these results support its continued use in that setting. PMID- 15753269 TI - Immigration, race/ethnicity, and social and economic factors as predictors of breastfeeding initiation. AB - OBJECTIVE: To determine the impact of immigration status as well as race/ethnicity and social and economic factors on breastfeeding initiation. DESIGN: Cohort. SETTING: Multisite group practice in eastern Massachusetts. PARTICIPANTS: One thousand eight hundred twenty-nine pregnant women prospectively followed up in Project Viva. MAIN OUTCOME MEASURE: Whether the participant breastfed her infant. RESULTS: The overall breastfeeding initiation rate was 83%. In multivariate models that included race/ethnicity and social, economic, and demographic factors, foreign-born women were more likely to initiate breastfeeding than US-born women (odds ratio [OR], 3.2 [95% confidence interval (CI), 2.0-5.2]). In models stratified by both race/ethnicity and immigration status, and further adjusted for whether the mother herself was breastfed as an infant and the mother's parents' immigration status, US-born and foreign-born black and Hispanic women initiated breastfeeding at rates at least as high as US born white women (US-born black vs US-born white women, OR, 1.2 [95% CI, 0.8 1.9], US-born Hispanic vs US-born white women, OR, 1.1 [95% CI, 0.6-1.9], foreign born black vs US-born white women, OR, 2.6 [95% CI, 1.1-6.0], and foreign-born Hispanic vs US-born white women, OR, 1.8 [95% CI, 0.7-4.8]). Calculations of predicted prevalences showed that, for example, the 2.6-fold increase in odds for the foreign-born black vs US-born white women translated to an increase in probability of approximately 1.4. Higher maternal education and household income also predicted higher initiation rates. CONCLUSIONS: Immigration status was strongly associated with increased breastfeeding initiation in this cohort, implying that cultural factors are important in the decision to breastfeed. Immigrants of all races/ethnicities initiated breastfeeding more often than their US-born counterparts. In addition, US-born minority groups initiated breastfeeding at rates at least as high as their white counterparts, likely due in part to high levels of education and income as well as to access to a medical care system that explicitly supports breastfeeding. PMID- 15753270 TI - Acculturation as a predictor of the onset of sexual intercourse among Hispanic and white teens. AB - OBJECTIVE: To investigate ethnic differences in onset of sexual intercourse among Hispanic/Mexican American and white adolescents based on acculturation. DESIGN/METHODS: Preprogram survey data from 7270 Hispanic or white teens in 7th to 12th grade involved in the Arizona Abstinence-Only Education Program were used to predict the probability of onset of sexual intercourse based on age, sex, family structure, program location, religiosity, free school lunch, grades, rural residence, acculturation, and ethnicity. Specific attention was given to the influence of acculturation among Hispanic teens. The primary language spoken by the respondents (English, Spanish, or both) was used as a proxy measure for acculturation. RESULTS: Hispanic youth were at a greater risk for experiencing onset of intercourse than white youth, while controlling for all other predictors (odds ratio [OR], 1.40 [95% confidence interval (CI), 1.21-1.63]). This risk was amplified for highly acculturated Hispanic teens (OR, 1.69 [95% CI, 1.43-1.99]). However, less acculturated Hispanic youth were actually less likely to have experienced first intercourse than white youth (OR, 0.59 [95% CI, 0.42-0.82]), English-speaking Hispanic youth (OR, 0.35 [95% CI, 0.25-0.49]), or bilingual Hispanic youth (OR, 0.45 [95% CI, 0.31-0.64]). CONCLUSIONS: Low acculturation emerges as a significant protective factor while controlling for other social and cultural factors, in spite of the increased risk of initiating sexual intercourse for Hispanic teens overall. Hispanic Spanish speakers were least likely to have initiated intercourse, while Hispanic English speakers were the most likely. PMID- 15753271 TI - Differences among states in the identification of autistic spectrum disorders. AB - OBJECTIVE: To explore variation among the 50 US states in the administrative prevalence of autistic spectrum disorders (ASDs) and factors associated with that variation. DESIGN: This survey study used secondary data from all 50 states obtained from the US Department of Education, Washington, DC, and the American Board of Pediatrics, Chapel Hill, NC. MAIN OUTCOME MEASURE: The outcome of interest was the proportion of children identified with ASD in the academic year 2000-2001. Linear regression was used to examine its association with education and health system characteristics. RESULTS: States ranged in the proportion of children diagnosed with ASD from 0.6 per 1000 to 4.6 per 1000 in 2000-2001. In adjusted analyses, education-related spending, the number of pediatricians in the state, and the number of school-based health centers in the state were positively associated with the administrative prevalence of ASD. CONCLUSIONS: Variation in the administrative prevalence of ASD is associated with education-related spending, which may be associated with better-trained educational staff who can recognize the problem, and more and better trained in-school specialists who can provide screening. It is also associated with the availability of health care resources. Increased access to pediatricians and school-based health centers may lead to improved recognition of ASD. Interstate variability in the identification of ASD should be taken into account when interpreting the results of prevalence studies based on administrative data and the associated system characteristics taken into account by policy makers working to improve the recognition of ASD. PMID- 15753272 TI - Violence perpetration among urban american Indian youth: can protection offset risk? AB - OBJECTIVE: To predict the likelihood of violence perpetration given various combinations of the most statistically salient risk and protective factors related to violence perpetration. DESIGN: Urban Indian Youth Health Survey, conducted from October 9, 1995, to March 30, 1998, consisting of 200 forced choice items exploring values, cultural identity, relationships, decision-making skills, and health and well-being. SETTING: Urban schools and an after-school youth development program at an urban American Indian center. PARTICIPANTS: Five hundred sixty-nine urban American Indian youth enrolled in grades 3 through 12. MAIN OUTCOME MEASURES: Violence perpetration dichotomized in 2 ways: (1) level of violence perpetration (ie, hitting someone 1-2 times in the past year vs picking fights, hitting repeatedly, participating in group fights, or shooting or stabbing someone in the past year) and (2) having shot and/or stabbed someone during the past year. RESULTS: In the final multivariate models with age as a covariate, most protective against violence perpetration were connections to school (odds ratio [OR], 0.17), positive affect (OR, 0.29), and peer prosocial behavior norms against violence (OR, 0.35). School connectedness (OR, 0.01) and positive affect (OR, 0.46) were also protective against shooting and/or stabbing someone, as was parental prosocial behavior norms against violence (OR, 0.23). The strongest risk factors for violence perpetration were substance use (OR, 2.60) and suicidal thoughts/behaviors (OR, 2.71); for shooting and/or stabbing, it was substance use (OR, 5.26). The likelihood of violence perpetration increased markedly (from 10% to 85%) as the exposure to risk factors increased and protective factors decreased. For shooting or stabbing someone, the probabilities ranged from 3% (0 risks and 3 protective factors) to 64% (1 risk and 0 protective factors). CONCLUSION: The dramatic reduction in the likelihood of violence involvement when risk was offset with protective factors in the probability profiles suggests the utility of a dual strategy of reducing risk while boosting protection. PMID- 15753273 TI - Effectiveness of oral dexamethasone in the treatment of moderate to severe pharyngitis in children. AB - OBJECTIVE: To determine the effectiveness of a single dose of oral dexamethasone in reducing the pain associated with moderate to severe pharyngitis in pediatric patients. DESIGN: Prospective, randomized, double-blind, placebo-controlled clinical trial. SETTING: Large, urban pediatric emergency department between March 2002 and November 2003. PATIENTS: Children aged 5 to 18 years with moderate to severe pharyngitis (odynophagia or dysphagia, moderate to severe pharyngeal erythema or swelling, and a McGrath Facial Affective Scale score of 0.75 or higher [scale 0.0-1.0]). INTERVENTIONS: Study patients were randomly assigned to receive 1 dose of either oral dexamethasone suspension (0.6 mg/kg with a maximum of 10 mg) or placebo of the same volume. All participants were tested for group A beta-hemolytic streptococcal pharyngitis and treated accordingly. Daily telephone follow-up was conducted until complete resolution of sore throat. MAIN OUTCOME MEASURES: Primary outcome variables included hours to initial relief of sore throat and time to the complete resolution of pain. Secondary outcome variables included changes in the McGrath Facial Affective Scale score at 24 and 48 hours, persistence of associated symptoms, use of anti-inflammatory or antipyretic medication, and subsequent use of medical resources for dehydration or pain. RESULTS: A convenience sample of 150 patients was randomized to receive either dexamethasone (n = 75) or placebo (n = 75). Twenty-five patients were lost to follow-up, leaving 125 patients available for data analysis; 57 received dexamethasone and 68 received placebo. Patients who received dexamethasone reported earlier onset of pain relief (9.2 vs 18.2 hours; P<.001), fewer hours to complete resolution of sore throat (30.3 vs 43.8 hours; P = .04), and larger changes in the McGrath Facial Affective Scale score in the first 24 hours (-0.58 vs -0.43; P = .002). Children who tested negative for group A beta-hemolytic streptococci had greater pain relief with dexamethasone compared with placebo (onset of pain relief, 8.7 vs 24 hours; P = .001), less time to complete resolution of sore throat (37.9 vs 70.8 hours; P = .006), and greater changes in the McGrath Facial Affective Scale score in the first 24 hours (-0.50 vs -0.21; P<.001). CONCLUSION: Children with moderate to severe pharyngitis had earlier onset of pain relief and shorter duration of sore throat when given oral dexamethasone. PMID- 15753274 TI - How do ethnicity and primary language spoken at home affect management practices and outcomes in children and adolescents with asthma? AB - BACKGROUND: Lower rates of preventive medication use and higher rates of hospitalization and emergency department use have been documented among Latino children and adolescents with asthma. However, little is known about how language barriers influence asthma management practices and outcomes. OBJECTIVE: To examine the effects of language on asthma management practices and asthma-related outcomes. DESIGN: Cross-sectional survey of asthma management practices, perceived efficacy, asthma knowledge, family functioning, and health-related quality of life in 405 white non-Latino, African American non-Latino, and Latino children and adolescents from English- and Spanish-speaking homes. RESULTS: Latino children and adolescents from Spanish-speaking homes had lower rates of goal setting and peak flow monitoring, poorer asthma knowledge, and greater negative family impact than white children and adolescents (P < .05 for all). Although Latino children and adolescents from English-speaking homes did worse than their non-Latino white peers, the decrements were modest and not statistically significant (P>.16 for all). Management practices and outcomes for non-Latino African American children and adolescents closely approximated those of white children and adolescents. CONCLUSIONS: Language barriers seem to contribute to poorer asthma management practices and knowledge among Latino children and adolescents. Efforts to increase knowledge in this group may enhance asthma self-care and limit the morbidity associated with asthma. PMID- 15753275 TI - Addressing obesity and other weight-related problems in youth. PMID- 15753276 TI - Lessons from planet health. PMID- 15753277 TI - The healthy immigrant effect: a greater understanding might help us improve the health of all children. PMID- 15753278 TI - Risk of mania with antidepressants. PMID- 15753279 TI - Sleep-disordered breathing and failure to thrive: research vs practice. PMID- 15753282 TI - Biography of Todd R. Klaenhammer. PMID- 15753281 TI - A role for the anaphase-promoting complex inhibitor Emi2/XErp1, a homolog of early mitotic inhibitor 1, in cytostatic factor arrest of Xenopus eggs. AB - Unfertilized vertebrate eggs are arrested in metaphase of meiosis II with high cyclin B/Cdc2 activity to prevent parthenogenesis. Until fertilization, exit from metaphase is blocked by an activity called cytostatic factor (CSF), which stabilizes cyclin B by inhibiting the anaphase-promoting complex (APC) ubiquitin ligase. The APC inhibitor early mitotic inhibitor 1 (Emi1) was recently found to be required for maintenance of CSF arrest. We show here that exogenous Emi1 is unstable in CSF-arrested Xenopus eggs and is destroyed by the SCF(betaTrCP) ubiquitin ligase, suggesting that endogenous Emi1, an apparent 44-kDa protein, requires a stabilizing factor. However, anti-Emi1 antibodies crossreact with native Emi2/Erp1/FBXO43, a homolog of Emi1 and conserved APC inhibitor. Emi2 is stable in CSF-arrested eggs, is sufficient to prevent CSF release, and is rapidly degraded in a Polo-like kinase 1-dependent manner in response to calcium-mediated egg activation. These results identify Emi2 as a candidate CSF maintenance protein. PMID- 15753283 TI - Hydrogen and energy flow as "sensed" by molecular genetics. PMID- 15753285 TI - Breakthroughs in the theory of Macdonald polynomials. PMID- 15753284 TI - Phytoplankton species richness scales consistently from laboratory microcosms to the world's oceans. AB - Species-area relationships have been observed for virtually all major groups of macroorganisms that have been studied to date but have not been explored for microscopic phytoplankton algae, which are the dominant producers in many freshwater and marine ecosystems. Our analyses of data from 142 different natural ponds, lakes, and oceans and 239 experimental ecosystems reveal a strong species area relationship with an exponent that is invariant across ecosystems that span >15 orders of magnitude in spatial extent. A striking result is that the species area relationship derived from small-scale experimental studies correctly scales up to natural aquatic ecosystems. These results significantly broaden our knowledge of the effects of island size on biodiversity and also confirm the relevance of experimentally derived data to the analysis and understanding of larger-scale ecological patterns. In addition, they confirm that patterns in microbial diversity are strongly consistent with those that have been repeatedly reported in the literature for macroorganisms. PMID- 15753286 TI - Biography of Francisco de la Cruz. PMID- 15753287 TI - Differential use of endoplasmic reticulum membrane for phagocytosis in J774 macrophages. AB - Sustained phagocytosis requires the continuous replacement of cell-surface membrane from intracellular sources. Depending on the nature of the engulfed particles, a variety of endocytic compartments have been demonstrated to contribute membranes needed for the formation of phagosomes. It has recently been reported that the endoplasmic reticulum (ER) can also fuse with the plasma membrane during phagocytosis [Gagnon, E., Duclos, S., Rondeau, C., Chevet, E., Cameron, P. H., Steele-Mortimer, O., Paiement, J., Bergeron, J. J. & Desjardins, M. (2002) Cell 110, 119-131]. However, there is currently no known mechanistic basis for this fusion process to occur. Here we report that direct ER-plasma membrane fusion during phagocytosis requires the ER resident soluble N ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein ERS24/Sec22b and that J774-macrophages react toward the challenge of large (3.0 microm) but not small (0.8-microm) particles by triggering this fusion mechanism, allowing them to access the most abundant endogenous membrane source in the cell, the ER. PMID- 15753288 TI - Characterization of functional and phenotypic changes in anti-Gag vaccine-induced T cell responses and their role in protection after HIV-1 infection. AB - Worldwide HIV-1 vaccine efforts are guided by the principle that HIV-specific T cell responses may provide protection from infection or delay overt disease. However, no clear correlates of T cell-mediated immune protection have been identified. Here, we examine in a HLA-B27(+) HIV seronegative vaccinee persistent HIV-specific vaccine-induced anti-Gag CD4(+) and CD8(+) T cell responses. Although these responses exhibited those characteristics (multifunctionality, appropriate memory phenotype, and targeting of epitopes associated with long-term nonprogression) predicted to correlate with protection from infection, the subject became HIV infected. After HIV infection, the vaccine-induced CD8(+) T cells expanded, but both CD4(+) and CD8(+) T cell responses acquired the functional and phenotypic patterns characteristic of chronic HIV infection. The virus quickly escaped the vaccine-induced T cell response, and the subject progressed more rapidly than expected for someone expressing the HLA-B27 allele. These data suggest that control of HIV by vaccine-elicited HIV-specific T cell responses may be difficult, even when the T cell response has those characteristics predicted to provide optimal protection. PMID- 15753289 TI - A spatially localized rhomboid protease cleaves cell surface adhesins essential for invasion by Toxoplasma. AB - Apicomplexan parasites cause serious human and animal diseases, the treatment of which requires identification of new therapeutic targets. Host-cell invasion culminates in the essential cleavage of parasite adhesins, and although the cleavage site for several adhesins maps within their transmembrane domains, the protease responsible for this processing has not been discovered. We have identified, cloned, and characterized the five nonmitochondrial rhomboid intramembrane proteases encoded in the recently completed genome of Toxoplasma gondii. Four T. gondii rhomboids (TgROMs) were active proteases with similar substrate specificity. TgROM1, TgROM4, and TgROM5 were expressed in the tachyzoite stage responsible for the disease, whereas TgROM2 and TgROM3 were expressed in the oocyst stage involved in transmission. Although both TgROM5 and TgROM4 localized to the cell surface in tachyzoites, TgROM5 was primarily at the posterior of the parasite, whereas adhesins were sequestered in internal micronemes. Upon microneme secretion, as occurs during invasion, the MIC2 adhesin was secreted to the apical end and translocated to the posterior, the site of cleavage, where it colocalized only with TgROM5. Moreover, only TgROM5 was able to cleave MIC adhesins in a cell-based assay, indicating that it likely provides the key protease activity necessary for invasion. T. gondii rhomboids have clear homologues in other apicomplexans including malaria; thus, our findings provide a model for studying invasion by this deadly pathogen and offer a target for therapeutic intervention. PMID- 15753290 TI - Genome-wide analysis of cAMP-response element binding protein occupancy, phosphorylation, and target gene activation in human tissues. AB - Hormones and nutrients often induce genetic programs via signaling pathways that interface with gene-specific activators. Activation of the cAMP pathway, for example, stimulates cellular gene expression by means of the PKA-mediated phosphorylation of cAMP-response element binding protein (CREB) at Ser-133. Here, we use genome-wide approaches to characterize target genes that are regulated by CREB in different cellular contexts. CREB was found to occupy approximately 4,000 promoter sites in vivo, depending on the presence and methylation state of consensus cAMP response elements near the promoter. The profiles for CREB occupancy were very similar in different human tissues, and exposure to a cAMP agonist stimulated CREB phosphorylation over a majority of these sites. Only a small proportion of CREB target genes was induced by cAMP in any cell type, however, due in part to the preferential recruitment of the coactivator CREB binding protein to those promoters. These results indicate that CREB phosphorylation alone is not a reliable predictor of target gene activation and that additional CREB regulatory partners are required for recruitment of the transcriptional apparatus to the promoter. PMID- 15753291 TI - Origin and evolution of the chicken leukocyte receptor complex. AB - In mammals, the cell surface receptors encoded by the leukocyte receptor complex (LRC) regulate the activity of T lymphocytes and B lymphocytes, as well as that of natural killer cells, and thus provide protection against pathogens and parasites. The chicken genome encodes many Ig-like receptors that are homologous to the LRC receptors. The chicken Ig-like receptor (CHIR) genes are members of a large monophyletic gene family and are organized into genomic clusters, which are in conserved synteny with the mammalian LRC. One-third of CHIR genes encode polypeptide molecules that contain both activating and inhibitory motifs. These genes are present in different phylogenetic groups, suggesting that the primordial CHIR gene could have encoded both types of motifs in a single molecule. In contrast to the mammalian LRC genes, the CHIR genes with similar function (inhibition or activation) are evolutionarily closely related. We propose that, in addition to recombination, single nucleotide substitutions played an important role in the generation of receptors with different functions. Structural models and amino acid analyses of the CHIR proteins reveal the presence of different types of Ig-like domains in the same phylogenetic groups, as well as sharing of conserved residues and conserved changes of residues between different CHIR groups and between CHIRs and LRCs. Our data support the notion that the CHIR gene clusters are regions homologous to the mammalian LRC gene cluster and favor a model of evolution by repeated processes of birth and death (expansion-contraction) of the Ig-like receptor genes. PMID- 15753292 TI - Lifelong elimination of hyperbilirubinemia in the Gunn rat with a single injection of helper-dependent adenoviral vector. AB - Crigler-Najjar syndrome is a recessively inherited disorder characterized by severe unconjugated hyperbilirubinemia caused by a deficiency of uridine diphospho-glucuronosyl transferase 1A1. Current therapy relies on phototherapy to prevent kernicterus, but liver transplantation presently is the only permanent cure. Gene therapy is a potential alternative, and recent work has shown that helper-dependent adenoviral (HD-Ad) vectors, devoid of all viral coding sequences, induce prolonged transgene expression and exhibit significantly less chronic toxicity than early-generation Ad vectors. We used a HD-Ad vector to achieve liver-restricted expression of human uridine diphospho-glucuronosyl transferase 1A1 in the Gunn rat, a model of the human disorder. Total plasma bilirubin levels were reduced from >5.0 mg/dl to <<1.4 mg/dl for >2 yr after a single i.v. administration of vector expressing the therapeutic transgene at a dose of 3 x 10(12) viral particles per kg. HPLC analysis of bile from treated rats showed the presence of bilirubin glucuronides at normal WT levels >2 yr after one injection of vector, and i.v. injection of bilirubins IIIalpha and XIIIalpha in the same animals revealed excess bilirubin-conjugating capacity. There was no significant elevation of liver enzymes (alanine aminotransferase) and only transient, moderate thrombocytopenia after injection of the vector. A clinically significant reduction in serum bilirubin was observed with a dose as low as 6 x 10(11) viral particles per kg. We conclude that complete, long-term correction of hyperbilirubinemia in the Gunn rat model of Crigler-Najjar syndrome can be achieved with one injection of HD-Ad vector and negligible chronic toxicity. PMID- 15753293 TI - Engineering of vault nanocapsules with enzymatic and fluorescent properties. AB - One of the central issues facing the emerging field of nanotechnology is cellular compatibility. Nanoparticles have been proposed for diagnostic and therapeutic applications, including drug delivery, gene therapy, biological sensors, and controlled catalysis. Viruses, liposomes, peptides, and synthetic and natural polymers have been engineered for these applications, yet significant limitations continue to prevent their use. Avoidance of the body's natural immune system, lack of targeting specificity, and the inability to control packaging and release are remaining obstacles. We have explored the use of a naturally occurring cellular nanoparticle known as the vault, which is named for its morphology with multiple arches reminiscent of cathedral ceilings. Vaults are 13-MDa ribonucleoprotein particles with an internal cavity large enough to sequester hundreds of proteins. Here, we report a strategy to target and sequester biologically active materials within the vault cavity. Attachment of a vault targeting peptide to two proteins, luciferase and a variant of GFP, resulted in their sequestration within the vault cavity. The targeted proteins confer enzymatic and fluorescent properties on the recombinant vaults, both of which can be detected by their emission of light. The modified vaults are compatible with living cells. The ability to engineer vault particles with designed properties and functionalities represents an important step toward development of a biocompatible nanocapsule. PMID- 15753294 TI - Architecture of recombination intermediates visualized by in-gel FRET of lambda integrase-Holliday junction-arm DNA complexes. AB - Lambda integrase (Int) mediates recombination between attachment sites on phage and Escherichia coli DNA. Int is assisted by accessory protein-induced DNA loops in bridging pairs of distinct "arm-type" and "core-type" DNA sites to form synapsed recombination complexes that subsequently recombine by means of a Holliday junction (HJ) intermediate. An in-gel FRET assay was developed and used to measure 15 distances between six points in two Int-HJ complexes containing arm DNA oligonucleotides, and 3D maps of these complexes were derived by distance geometry calculations. The maps reveal unexpected positions for the arm-type DNAs relative to core sites on the HJ and a new Int conformation in the HJ tetramer. The results show how the position of arm DNAs determines the bias of catalytic activities responsible for directional resolution, provide insights into the organization of Int higher-order complexes, and lead to models of the structure of the full HJ recombination intermediates. PMID- 15753295 TI - Magnetic self-assembly of three-dimensional surfaces from planar sheets. AB - This report describes the spontaneous folding of flat elastomeric sheets, patterned with magnetic dipoles, into free-standing, 3D objects that are the topological equivalents of spherical shells. The path of the self-assembly is determined by a competition between mechanical and magnetic interactions. The potential of this strategy for the fabrication of 3D electronic devices is demonstrated by generating a simple electrical circuit surrounding a spherical cavity. PMID- 15753296 TI - Cap-binding protein 1-mediated and eukaryotic translation initiation factor 4E mediated pioneer rounds of translation in yeast. AB - Nonsense-mediated mRNA decay (NMD) in mammalian cells is restricted to newly synthesized mRNA that is bound at the 5' cap by the major nuclear cap-binding complex and at splicing-generated exon-exon junctions by exon junction complexes. This messenger ribonucleoprotein has been called the pioneer translation initiation complex and, accordingly, NMD occurs as a consequence of nonsense codon recognition during a pioneer round of translation. Here, we characterize the nature of messenger ribonucleoprotein that is targeted for NMD in Saccharomyces cerevisiae. Data indicate that NMD targets both cap-binding complex (Cbc)1p- and eukaryotic translation initiation factor (eIF)4E-bound mRNAs, unlike in mammalian cells, where NMD does not detectably target eIF4E-bound mRNA. First, intron-containing pre-mRNAs in yeast are detectably bound by either Cbc1p, or, unlike in mammalian cells, eIF4E, indicating that mRNAs can be derived from either Cbc1p- or eIF4E-bound pre-mRNAs. Second, the ratio of nonsense-containing Cbc1p-bound mRNA to nonsense-free Cbc1p-bound mRNA, which was < 0.4 for those mRNAs tested here, is essentially identical to the ratio of the corresponding nonsense-containing eIF4E-bound mRNA to nonsense-free eIF4E-bound mRNA, and both ratios increase in cells treated with the translational inhibitor cycloheximide (CHX). These data, together with data presented here and elsewhere showing that Cbc1p-bound transcripts are precursors to eIF4E-bound transcripts, demonstrate that Cbc1p-bound mRNA is targeted for NMD. In support of the idea that eIF4E bound mRNA is also targeted for NMD, eIF4E-bound mRNA is targeted for NMD in strains that lack Cbc1p. These results suggest that both Cbc1p- and eIF4E mediated pioneer rounds of translation occur in yeast. PMID- 15753299 TI - Viscosity-dependent inertial spectra of the Burgers and Korteweg-deVries-Burgers equations. AB - We show that the inertial range spectrum of the Burgers equation has a viscosity dependent correction at any wave number when the viscosity is small but not zero. We also calculate the spectrum of the Korteweg-deVries-Burgers equation and show that it can be partially mapped onto the inertial spectrum of a Burgers equation with a suitable effective diffusion coefficient. These results are significant for the understanding of turbulence. PMID- 15753298 TI - Osteocalcin protein sequences of Neanderthals and modern primates. AB - We report here protein sequences of fossil hominids, from two Neanderthals dating to approximately 75,000 years old from Shanidar Cave in Iraq. These sequences, the oldest reported fossil primate protein sequences, are of bone osteocalcin, which was extracted and sequenced by using MALDI-TOF/TOF mass spectrometry. Through a combination of direct sequencing and peptide mass mapping, we determined that Neanderthals have an osteocalcin amino acid sequence that is identical to that of modern humans. We also report complete osteocalcin sequences for chimpanzee (Pan troglodytes) and gorilla (Gorilla gorilla gorilla) and a partial sequence for orangutan (Pongo pygmaeus), all of which are previously unreported. We found that the osteocalcin sequences of Neanderthals, modern human, chimpanzee, and orangutan are unusual among mammals in that the ninth amino acid is proline (Pro-9), whereas most species have hydroxyproline (Hyp-9). Posttranslational hydroxylation of Pro-9 in osteocalcin by prolyl-4-hydroxylase requires adequate concentrations of vitamin C (l-ascorbic acid), molecular O(2), Fe(2+), and 2-oxoglutarate, and also depends on enzyme recognition of the target proline substrate consensus sequence Leu-Gly-Ala-Pro-9-Ala-Pro-Tyr occurring in most mammals. In five species with Pro-9-Val-10, hydroxylation is blocked, whereas in gorilla there is a mixture of Pro-9 and Hyp-9. We suggest that the absence of hydroxylation of Pro-9 in Pan, Pongo, and Homo may reflect response to a selective pressure related to a decline in vitamin C in the diet during omnivorous dietary adaptation, either independently or through the common ancestor of these species. PMID- 15753297 TI - Serine/arginine-rich protein-dependent suppression of exon skipping by exonic splicing enhancers. AB - The 5' and 3' splice sites within an intron can, in principle, be joined to those within any other intron during pre-mRNA splicing. However, exons are joined in a strict 5' to 3' linear order in constitutively spliced pre-mRNAs. Thus, specific mechanisms must exist to prevent the random joining of exons. Here we report that insertion of exon sequences into an intron can inhibit splicing to the downstream 3' splice site and that this inhibition is independent of intron size. The exon sequences required for splicing inhibition were found to be exonic enhancer elements, and their inhibitory activity requires the binding of serine/arginine rich splicing factors. We conclude that exonic enhancers can act as barriers to prevent exon skipping and thereby may play a key role in ensuring the correct 5' to 3' linear order of exons in spliced mRNA. PMID- 15753300 TI - Homoserine and asparagine are host signals that trigger in planta expression of a pathogenesis gene in Nectria haematococca. AB - Some pathogenesis-related genes are expressed in fungi only when the pathogen is in the host, but the host signals that trigger these gene expressions have not been identified. Virulent Nectria haematococca infects pea plants and requires either pelA, which is induced by pectin, or pelD, which is induced only in planta. However, the host signal(s) that trigger pelD expression was unknown. Here we report the isolation of the host signals and identify homoserine and asparagine, two free amino acids found in uniquely high levels in pea seedlings, as the pelD-inducing signals. N. haematococca has evolved a mechanism to sense the host tissue environment by using the high levels of two free amino acids in this plant, thereby triggering the expression of pelD to assist the pathogenic process. PMID- 15753301 TI - The protooncogene MYC can break B cell tolerance. AB - The protooncogene MYC has been implicated in both the proliferation and programmed cell death of lymphoid cells, and in the genesis of lymphoid tumors. Here, we report that overexpression of MYC, as found in many lymphomas, can break immune tolerance. Mice that would otherwise be tolerant to a transgenic autoantigen mounted an immune response to the antigen if MYC was vigorously expressed in the B cell lineage. The responsive B cells converted to an activated phenotype and produced copious amounts of autoantibody that engendered immune complex disease of the kidney. MYC was required to both establish and maintain the breach of tolerance. These effects may be due to the ability of MYC to serve as a surrogate for cytokines. We found that the gene could mimic the effects of cytokines on both B cell proliferation and survival and, indeed, was required for those effects. These findings demonstrate a critical role for MYC in the response of B cells to antigen and expand the potential contributions of MYC to the genesis of lymphomas. PMID- 15753302 TI - Nitroaspirin corrects immune dysfunction in tumor-bearing hosts and promotes tumor eradication by cancer vaccination. AB - Active suppression of tumor-specific T lymphocytes can limit the immune-mediated destruction of cancer cells. Of the various strategies used by tumors to counteract immune attacks, myeloid suppressors recruited by growing cancers are particularly efficient, often resulting in the induction of systemic T lymphocyte dysfunction. We have previously shown that the mechanism by which myeloid cells from tumor-bearing hosts block immune defense strategies involves two enzymes that metabolize L-arginine: arginase and nitric oxide (NO) synthase. NO-releasing aspirin is a classic aspirin molecule covalently linked to a NO donor group. NO aspirin does not possess direct antitumor activity. However, by interfering with the inhibitory enzymatic activities of myeloid cells, orally administered NO aspirin normalized the immune status of tumor-bearing hosts, increased the number and function of tumor-antigen-specific T lymphocytes, and enhanced the preventive and therapeutic effectiveness of the antitumor immunity elicited by cancer vaccination. Because cancer vaccines and NO aspirin are currently being investigated in independent phase I/II clinical trials, these findings offer a rationale to combine these treatments in subjects with advanced neoplastic diseases. PMID- 15753303 TI - Midgut epithelial responses of different mosquito-Plasmodium combinations: the actin cone zipper repair mechanism in Aedes aegypti. AB - In vivo responses of midgut epithelial cells to ookinete invasion of three different vector-parasite combinations, Aedes aegypti-Plasmodium gallinaceum, Anopheles stephensi-Plasmodium berghei, and A. stephensi-P. gallinaceum, were directly compared by using enzymatic markers and immunofluorescence stainings. Our studies indicate that, in A. aegypti and A. stephensi ookinetes traverse the midgut via an intracellular route and inflict irreversible damage to the invaded cells. These two mosquito species differ, however, in their mechanisms of epithelial repair. A. stephensi detaches damaged cells by an actin-mediated budding-off mechanism when invaded by either P. berghei or P. gallinaceum. In A. aegypti, the midgut epithelium is repaired by a unique actin cone zipper mechanism that involves the formation of a cone-shaped actin aggregate at the base of the cell that closes sequentially, expelling the cellular contents into the midgut lumen as it brings together healthy neighboring cells. Invasion of A. stephensi by P. berghei induced expression of nitric oxide synthase and peroxidase activities, which mediate tyrosine nitration. These enzymes and nitrotyrosine, however, were not induced in the other two vector-parasite combinations examined. These studies indicate that the epithelial responses of different mosquito-parasite combinations are not universal. The implications of these observations to validate animal experimental systems that reflect the biology of natural vectors of human malarias are discussed. PMID- 15753304 TI - A copper(I) protein possibly involved in the assembly of CuA center of bacterial cytochrome c oxidase. AB - Sco1 and Cox17 are accessory proteins required for the correct assembly of eukaryotic cytochrome c oxidase. At variance with Sco1, Cox17 orthologs are found only in eukaryotes. We browsed bacterial genomes to search proteins functionally equivalent to Cox17, and we identified a class of proteins of unknown function displaying a conserved gene neighborhood to bacterial Sco1 genes, all sharing a potential metal binding motif H(M)X10MX21HXM. Two members of this group, DR1885 from Deinococcus radiodurans and CC3502 from Caulobacter crescentus, were expressed, and their interaction with copper was investigated. The solution structure and extended x-ray absorption fine structure data on the former protein reveal that the protein binds copper(I) through a histidine and three Mets in a cupredoxin-like fold. The surface location of the copper-binding site as well as the type of coordination are well poised for metal transfer chemistry, suggesting that DR1885 might transfer copper, taking the role of Cox17 in bacteria. On the basis of our results, a possible pathway for copper delivery to the Cu(A) center in bacteria is proposed. PMID- 15753305 TI - Site-directed spin labeling reveals a conformational switch in the phosphorylation domain of smooth muscle myosin. AB - We have used site-directed spin labeling and EPR spectroscopy to detect structural changes within the regulatory light chain (RLC) of smooth muscle myosin upon phosphorylation. Smooth muscle contraction is activated by phosphorylation of S19 on RLC, but the structural basis of this process is unknown. There is no crystal structure containing a phosphorylated RLC, and there is no crystal structure for the N-terminal region of any RLC. Therefore, we have prepared single-Cys mutations throughout RLC, exchanged each mutant onto smooth muscle heavy meromyosin, verified normal regulatory function, and used EPR to determine dynamics and solvent accessibility at each site. A survey of spin-label sites throughout the RLC revealed that only the N-terminal region (first 24 aa) shows a significant change in dynamics upon phosphorylation, with most of the first 17 residues showing an increase in rotational amplitude. Therefore, we focused on this N-terminal region. Additional structural information was obtained from the pattern of oxygen accessibility along the sequence. In the absence of phosphorylation, little or no periodicity was observed, suggesting a lack of secondary structural order in this region. However, phosphorylation induced a strong helical pattern (3.6-residue periodicity) in the first 17 residues, while increasing accessibility throughout the first 24 residues. We have identified a domain within RLC, the N-terminal phosphorylation domain, in which phosphorylation increases helical order, internal dynamics, and accessibility. These results support a model in which this disorder-to-order transition within the phosphorylation domain results in decreased head-head interactions, activating myosin in smooth muscle. PMID- 15753306 TI - Foxp3+ CD25- CD4 T cells constitute a reservoir of committed regulatory cells that regain CD25 expression upon homeostatic expansion. AB - Expression of the IL-2 receptor alpha chain (CD25) by peripheral CD4 T cells follows cellular activation. However, CD25 expression by CD4 cells is widely used as a marker to identify regulatory T cells (T(R)), although cells with regulatory properties are also found in the CD4+CD25- subset. By using in vivo functional assays and Foxp3 expression as a faithful marker of T(R) differentiation, we have evaluated the requirements for CD25 expression by peripheral T(R). We first show that in vivo depletion of CD25+ cells prevents the development of spontaneous encephalomyelitis in recombination-activating gene (RAG)-deficient anti-myelin basic protein T cell antigen receptor (TCR) transgenic mice, and allows disease induction in otherwise healthy RAG-competent transgenic mice. Similar treatment in normal thymectomized animals is followed by the fast recovery of a normal number of CD25+ T(R). Consistently, Foxp3-expressing T(R) encompassed in the CD25 cell population convert to CD25+ after homeostatic expansion and are selectable by IL-2 in vitro. Surface expression of CD25 on T(R) is controlled by the activity of conventional CD4 cells and is fully labile because it can be lost and regained without affecting the functional potential of the cells. These findings reveal that Foxp3-expressing CD25- cells constitute a peripheral reservoir of differentiated T(R), recruited to the CD25+ pool upon homeostatic expansion and/or activation. This analysis, together with the notion that physiological commitment of T(R) takes place exclusively in the thymus should help for the interpretation of experiments assessing peripheral T(R) differentiation from naive CD4 T cells, defined as CD25-. PMID- 15753308 TI - Disease-associated mutations cause premature oligomerization of myelin proteolipid protein in the endoplasmic reticulum. AB - Pelizaeus-Merzbacher disease (PMD) is a dysmyelinating disease caused by mutations, deletions, or duplications of the proteolipid protein (PLP) gene. Mutant forms of PLP are retained in the endoplasmic reticulum (ER), and the resulting accumulation of mutant protein is thought to be a direct cause of oligodendrocyte cell death, which is the primary clinical feature of PMD. The molecular mechanisms underlying the toxicity of mutant PLP are however currently unknown. We report here that PMD-linked mutations of PLP are associated with the accelerated assembly of the protein into stable homooligomers that resemble mature, native PLP. Thus although WT PLP forms stable oligomers after an extended maturation period, most likely at the cell surface, mutant forms of PLP rapidly assemble into such oligomers at the ER. Using PLP mutants associated with diseases of varying severity, we show that the formation of stable oligomers correlates with the development of PMD. Based on these findings, we propose that the premature oligomerization of PLP in the ER of oligodendrocytes contributes to the pathology of PMD. PMID- 15753309 TI - Simulation of dynamical properties of normal and superfluid helium. AB - The formation of a superfluid when (4)He is cooled below the characteristic lambda transition temperature is accompanied by intricate quantum mechanical phenomena, including the emergence of a Bose condensate. A combination of path integral and semiclassical techniques is used to calculate the single-particle velocity autocorrelation function across the normal-to-superfluid transition. We find that the inclusion of particle exchange alters qualitatively the shape of the correlation function below the characteristic transition temperature but has no noticeable effect on the dynamics in the normal phase. The incoherent structure factor extracted from the velocity autocorrelation function is in very good agreement with neutron scattering data, reproducing the width, height, frequency shift, and asymmetry of the curves, as well as the observed increase in peak height characteristic of the superfluid phase. Our simulation demonstrates that the peak enhancement observed in the neutron scattering experiments below the transition temperature arises exclusively from particle exchange, illuminating the role of Bose-statistical effects on the dynamics of the quantum liquid. PMID- 15753307 TI - Viral genetic determinants for thrips transmission of Tomato spotted wilt virus. AB - Tomato spotted wilt virus (TSWV) is transmitted exclusively by thrips in nature. A reassortment-based viral genetic system was used to map transmissibility by thrips to the medium (M) RNA of TSWV. To locate determinants of thrips transmission in the M RNA, 30 single-lesion isolates (SLIs) were generated from a single TSWV isolate that was inefficiently transmitted by thrips. Three of the 30 SLIs were transmitted by thrips, and 27 were not. Sequence analysis of the M RNA, thrips transmissibility assays, G(C) protein analysis, and transmission electron microscopic studies revealed that a specific nonsynonymous mutation (C1375A) in the G(N)/G(C) ORF of the M RNA resulted in the loss of thrips transmissibility without inhibition of virion assembly. This was in contrast to other nontransmissible SLIs, which had frameshift and/or nonsense mutations in the G(N)/G(C) ORF but were defective in virion assembly. The G(C) glycoprotein was detectable in the C1375A mutants but not in the frameshift or nonsense mutants. We report a specific viral determinant associated with virus transmission by thrips. In addition, the loss of transmissibility was associated with the accumulation of defective haplotypes in the population, which are not transmissible by thrips, rather than with the presence of a dominant haplotype that is inefficiently transmitted by thrips. These results also indicate that the glycoproteins may not be required for TSWV infection of plant hosts but are required for transmissibility by thrips. PMID- 15753310 TI - Implications of an antiparallel dimeric structure of nonphosphorylated STAT1 for the activation-inactivation cycle. AB - IFN-gamma treatment of cells leads to tyrosine phosphorylation of signal transducer and activator of transcription (STAT) 1 followed by dimerization through a reciprocal Src homology 2-phosphotyrosine interaction near the -COOH end of each monomer, forming a parallel structure that accumulates in the nucleus to drive transcription. Prompt dephosphorylation and return to the cytoplasm completes the activation-inactivation cycle. Nonphosphorylated STATs dimerize, and a previously described interface between N-terminal domain (ND) dimers has been implicated in this dimerization. A new crystal structure of nonphosphorylated STAT1 containing the ND dimer has two possible configurations for the body of STAT1, one of which is antiparallel. In this antiparallel structure, the Src homology 2 domains are at opposite ends of the dimer, with the coiled:coil domain of one monomer interacting reciprocally with the DNA-binding domain of its partner. Here, we find that mutations in either the coiled:coil/DNA binding domain interface or the ND dimer interface block dimerization of nonphosphorylated molecules and cause a resistance to dephosphorylation in vivo and resistance to a tyrosine phosphatase in vitro. We conclude that a parallel STAT1 phosphodimer not bound to DNA most likely undergoes a conformational rearrangement (parallel to antiparallel) to present the phosphotyrosine efficiently for dephosphorylation. PMID- 15753311 TI - A binding mechanism in protein-nucleotide interactions: implication for U1A RNA binding. AB - We present a close electronic view of the protein-base interface for the N terminal domain of the human protein U1A. Combining accurate mixed quantum mechanics/molecular mechanics techniques and protein structure prediction methods, we provide a detailed electronic structure description of the protein RNA stacking interactions. Our analysis indicates the evolution of the protein structure optimizing the interaction between Asp-92 and the RNA bases. The results show a direct coupling of the C-terminal tail and Asp-92, providing a direct rationalization of the experimentally determined role of the C-terminal domain in RNA binding. Here, we propose a mechanism where a protein side chain, with a delocalized electronic pi system, assists in the nucleotide binding. The binding mechanism involves a short-range interaction of the side chain with the nucleotide base and an electronic long-range interaction through a sandwich stacking motif. The structural motif of the binding mechanism is observed in similar protein-RNA interactions and in various protein-ATP-binding sites. PMID- 15753312 TI - Reduced intercellular coupling leads to paradoxical propagation across the Purkinje-ventricular junction and aberrant myocardial activation. AB - Ventricular tachycardia is a common heart rhythm disorder and a frequent cause of sudden cardiac death. Aberrant cell-cell coupling through gap junction channels, a process termed gap junction remodeling, is observed in many of the major forms of human heart disease and is associated with increased arrhythmic risk in both humans and in animal models. Genetically engineered mice with cardiac-restricted knockout of Connexin43, the major cardiac gap junctional protein, uniformly develop sudden cardiac death, although a detailed electrophysiological understanding of their profound arrhythmic propensity is unclear. Using voltage sensitive dyes and high resolution optical mapping techniques, we found that uncoupling of the ventricular myocardium results in ectopic sites of ventricular activation. Our data indicate that this behavior reflects alterations in source sink relationships and paradoxical conduction across normally quiescent Purkinje ventricular muscle junctions. The aberrant activation profiles are associated with wavefront collisions, which in the setting of slow conduction may account for the highly arrhythmogenic behavior of Connexin43-deficient hearts. Thus, the extent of gap junction remodeling in diseased myocardium is a critical determinant of cardiac excitation patterns and arrhythmia susceptibility. PMID- 15753313 TI - Meiotic and epigenetic defects in Dnmt3L-knockout mouse spermatogenesis. AB - The production of mature germ cells capable of generating totipotent zygotes is a highly specialized and sexually dimorphic process. The transition from diploid primordial germ cell to haploid spermatozoa requires genome-wide reprogramming of DNA methylation, stage- and testis-specific gene expression, mitotic and meiotic division, and the histone-protamine transition, all requiring unique epigenetic control. Dnmt3L, a DNA methyltransferase regulator, is expressed during gametogenesis, and its deletion results in sterility. We found that during spermatogenesis, Dnmt3L contributes to the acquisition of DNA methylation at paternally imprinted regions, unique nonpericentric heterochromatic sequences, and interspersed repeats, including autonomous transposable elements. We observed retrotransposition of an LTR-ERV1 element in the DNA from Dnmt3L-/- germ cells, presumably as a result of hypomethylation. Later in development, in Dnmt3L-/- meiotic spermatocytes, we detected abnormalities in the status of biochemical markers of heterochromatin, implying aberrant chromatin packaging. Coincidentally, homologous chromosomes fail to align and form synaptonemal complexes, spermatogenesis arrests, and spermatocytes are lost by apoptosis and sloughing. Because Dnmt3L expression is restricted to gonocytes, the presence of defects in later stages reveals a mechanism whereby early genome reprogramming is linked inextricably to changes in chromatin structure required for completion of spermatogenesis. PMID- 15753314 TI - Cytosolic abscisic acid activates guard cell anion channels without preceding Ca2+ signals. AB - The phytohormone abscisic acid (ABA) reports on the water status of the plant and induces stomatal closure. Guard cell anion channels play a central role in this response, because they mediate anion efflux, and in turn, cause a depolarization induced K+ release. We recorded early steps in ABA signaling, introducing multibarreled microelectrodes in guard cells of intact plants. Upon external ABA treatment, anion channels transiently activated after a lag phase of approximately 2 min. As expected for a cytosolic ABA receptor, iontophoretic ABA loading into the cytoplasm initiated a rise in anion current without delay. These ABA responses could be elicited repetitively at resting and at largely depolarized potentials (e.g., 0 mV), ruling out signal transduction by means of hyperpolarization-activated calcium channels. Likewise, ABA stimulation did not induce a rise in the cytosolic free-calcium concentration. However, the presence of approximately 100 nM background Ca2+ was required for anion channel function, because the action of ABA on anion channels was repressed after loading of the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetate. The chain of events appears very direct, because none of the tested putative ABA-signaling intermediates (inositol 1,4,5 trisphosphate, inositol hexakisphosphate, nicotinic acid adenine dinucleotide phosphate, and cyclic ADP-ribose), could mimic ABA as anion channel activator. In patch-clamp experiments, cytosolic ABA also evoked anion current transients carried by R- and S-type anion channels. The response was dose-dependent with half-maximum activation at 2.6 microM ABA. Our studies point to an ABA pathway initiated by ABA binding to a cytosolic receptor that within seconds activates anion channels, and in turn, leads to depolarization of the plasma membrane. PMID- 15753315 TI - Mechanism of phagolysosome biogenesis block by viable Mycobacterium tuberculosis. AB - Live Mycobacterium tuberculosis persists in macrophage phagosomes by interfering with phagolysosome biogenesis. Here, using four-dimensional microscopy and in vitro assays, we report the principal difference between phagosomes containing live and dead mycobacteria. Phosphatidylinositol 3-phosphate (PI3P), a membrane trafficking regulatory lipid essential for phagosomal acquisition of lysosomal constituents, is retained on phagosomes harboring dead mycobacteria but is continuously eliminated from phagosomes with live bacilli. We show that the exclusion of PI3P from live mycobacterial phagosomes can be only transiently reversed by Ca2+ fluxes, and that live M. tuberculosis secretes a lipid phosphatase, SapM, that hydrolyzes PI3P, inhibits phagosome-late endosome fusion in vitro, and contributes to inhibition of phagosomal maturation. PMID- 15753316 TI - Calling for help is independently modulated by brain systems underlying goal directed behavior and threat perception. AB - In primates, during times of need, calling for help is a universal experience. Calling for help recruits social support and promotes survival. However, calling for help also can attract predators, and it is adaptive to inhibit calls for help when a potential threat is perceived. Based on this, we hypothesized that individual differences in calling for help would be related to the activity of brain systems that mediate goal-directed behavior and the detection of threat. By using high-resolution positron emission tomography in rhesus monkeys undergoing social separation, we demonstrate that increased [18F]-fluoro-2-deoxy-D-glucose uptake in the right dorsolateral prefrontal cortex and decreased uptake in the amygdala independently predict individual differences in calling for help. When taken together, these two regions account for 76% of the variance in calling for help. This result suggests that the drive for affiliation and the perception of threat determine the intensity of an individual's behavior during separation. These findings in monkeys are relevant to humans and provide a conceptual neural framework to understand individual differences in how primates behave when in need of social support. PMID- 15753317 TI - The Anopheles gambiae detoxification chip: a highly specific microarray to study metabolic-based insecticide resistance in malaria vectors. AB - Metabolic pathways play an important role in insecticide resistance, but the full spectra of the genes involved in resistance has not been established. We constructed a microarray containing unique fragments from 230 Anopheles gambiae genes putatively involved in insecticide metabolism [cytochrome P450s (P450s), GSTs, and carboxylesterases and redox genes, partners of the P450 oxidative metabolic complex, and various controls]. We used this detox chip to monitor the expression of the detoxifying genes in insecticide resistant and susceptible An. gambiae laboratory strains. Five genes were strongly up-regulated in the dichlorodiphenyltrichloroethane-resistant strain ZAN/U. These genes included the GST GSTE2, which has previously been implicated in dichlorodiphenyltrichloroethane resistance, two P450s, and two peroxidase genes. GSTE2 was also elevated in the pyrethroid-resistant RSP strain. In addition, the P450 CYP325A3, belonging to a class not previously associated with insecticide resistance, was expressed at statistically higher levels in this strain. The applications of this detox chip and its potential contribution to malaria vector insecticide resistance management programs are discussed. PMID- 15753318 TI - De novo generation of antigen-specific CD4+CD25+ regulatory T cells from human CD4+CD25- cells. AB - Antigen-specificity is a hallmark of adaptive T cell-mediated immune responses. CD4+CD25+FOXP3+ regulatory T cells (T(R)) also require activation through the T cell receptor for function. Although these cells require antigen-specific activation, they are generally able to suppress bystander T cell responses once activated. This raises the possibility that antigen-specific T(R) may be useful therapeutically by localizing generalized suppressive activity to tissues expressing select target antigens. Here, we demonstrate that T(R) specific for particular peptide-MHC complexes can be generated from human CD4+CD25- T cells in vitro and isolated by using HLA class II tetramers. Influenza hemagglutinin epitopes were used to generate hemagglutinin-specific T(R), which required cognate antigen for activation but which subsequently suppressed noncognate bystander T cell responses as well. These findings have implications for the generation of therapeutic regulatory T cells in disease, and also suggest an important mechanism by which T cells may be regulated at the site of inflammation. PMID- 15753319 TI - Genetically stable and fully effective smallpox vaccine strain constructed from highly attenuated vaccinia LC16m8. AB - A highly attenuated LC16m8 (m8) smallpox vaccine has been licensed in Japan because of its extremely low neurovirulence profile, which is comparable to that of replication incompetent strains of vaccinia virus. From 1973 to 1975, m8 was administrated to >100,000 infants where it induced levels of immunity similar to that of the originating Lister strain, without any serious side effects. Recently, we observed that m8 reverts spontaneously to large plaque forming clones that possess virulence equivalent to that of LC16mO, a parental virus strain of m8. Here, we report that the B5R gene is responsible for the reversion, and that we could construct a more genetically stable virus by deleting B5R from m8. The protective immunogenicity of the vaccine candidate proved to be equivalent to that of the U.S.-licensed product Dryvax, and much superior to modified vaccinia Ankara in a mouse model. Furthermore, the vaccine strain never elicited any symptoms in severe combined immunodeficiency disease mice, even at a dose 1,000-fold greater than that used in the immune protection experiments, which is in contrast to the lethal pathogenicity induced by Dryvax inoculation of severe combined immunodeficiency disease mice. Our results suggest that this vaccine strain is a good candidate as a suitable smallpox vaccine and a vector virus, and that B5R is not essential for protective immunity against smallpox. PMID- 15753320 TI - Role of structural determinants in folding of the sandwich-like protein Pseudomonas aeruginosa azurin. AB - An invariant substructure that forms two interlocked pairs of neighboring beta strands occurs in essentially all known sandwich-like proteins. Eight conserved positions in these strands were recently shown to act as structural determinants. To test whether the residues at these invariant positions are conserved for mechanistic (i.e., part of folding nucleus) or energetic (i.e., governing native state stability) reasons, we characterized the folding behavior of eight point mutated variants of the sandwich-like protein Pseudomonas aeruginosa apo-azurin. We find a simple relationship among the conserved positions: half of the residues form native-like interactions in the folding transition state, whereas the others do not participate in the folding nucleus but govern high native-state stability. Thus, evolutionary preservation of these specific positions gives both mechanistic and energetic advantages to members of the sandwich-like protein family. PMID- 15753321 TI - Risk preference instability across institutions: a dilemma. AB - In this article we use laboratory experiments to ask a fundamental question: Do individuals behave as if their risk preferences are stable across institutions? In particular, we study the decisions of cash-motivated subjects in the repeated play of three different institutions: a value elicitation procedure for the sale of a risky asset, an English clock auction for the sale of a risky asset, and a first-price auction for the purchase of a riskless asset. We first do a simple categorical comparison of each subject's risk preferences across tasks by comparing the individual's decisions with an expected value maximizer. All subjects acted as if they were risk-loving in the English clock auctions and risk averse in the first-price auctions. In the Becker, Degroot, and Marschack procedure, behavior was split between risk-loving and risk-averse bidding. For each institution we also estimate an individual's risk coefficient. We test the hypotheses that for the same individuals the estimated risk coefficient across institutions is the same. We find that these estimates are statistically different. PMID- 15753322 TI - Convergent regulation of skeletal muscle Ca2+ channels by dystrophin, the actin cytoskeleton, and cAMP-dependent protein kinase. AB - The skeletal muscle L-type Ca2+ channel (Ca(V)1.1), which is responsible for initiating muscle contraction, is regulated by phosphorylation by cAMP-dependent protein kinase (PKA) in a voltage-dependent manner that requires direct physical association between the channel and the kinase mediated through A-kinase anchoring proteins (AKAPs). The role of the actin cytoskeleton in channel regulation was investigated in skeletal myocytes cultured from wild-type mice, mdx mice that lack the cytoskeletal linkage protein dystrophin, and a skeletal muscle cell line, 129 CB3. Voltage dependence of channel activation was shifted positively, and potentiation was greatly diminished in mdx myocytes and in 129 CB3 cells treated with the microfilament stabilizer phalloidin. Voltage-dependent potentiation by strong depolarizing prepulses was reduced in mdx myocytes but could be restored by positively shifting the stimulus potentials to compensate for the positive shift in the voltage dependence of gating. Inclusion of PKA in the pipette caused a negative shift in the voltage dependence of activation and restored voltage-dependent potentiation in mdx myocytes. These results show that skeletal muscle Ca2+ channel activity and voltage-dependent potentiation are controlled by PKA and microfilaments in a convergent manner. Regulation of Ca2+ channel activity by hormones and neurotransmitters that use the PKA signal transduction pathway may interact in a critical way with the cytoskeleton and may be impaired by deletion of dystrophin, contributing to abnormal regulation of intracellular calcium concentrations in dystrophic muscle. PMID- 15753323 TI - Enhanced cocaine responsiveness and impaired motor coordination in metabotropic glutamate receptor subtype 2 knockout mice. AB - Extensive pharmacological studies have recently emerged indicating that group 2 metabotropic glutamate receptors (mGluRs) comprising mGluR2 and mGluR3 subtypes are associated with several neurological and psychiatric disorders. mGluR2 is widely distributed both presynaptically and postsynaptically in a variety of neuronal cells, but the physiological role of mGluR2 in brain function is poorly understood. This investigation involves a comprehensive behavioral analysis of mGluR2-/- knockout (KO) mice to explore the physiological role of mGluR2 in brain function. Although, under general observation, mGluR2-/- KO mice appeared to have no behavioral abnormalities, they exhibited several lines of behavioral alterations in the enforcing and defined behavioral tests. They showed a significant increase in locomotor sensitization and conditioned place preference in association with repeated cocaine administration, indicating that mGluR2 contributes to behavioral responses implicated in reinforcement and addiction of cocaine. Upon in vivo microdialysis analysis after cocaine administration, not only did extracellular levels of dopamine increase but also the response pattern of glutamate release markedly changed in the nucleus accumbens of mGluR2-/- KO mice. The mGluR2-/- KO mice also showed significant impairment in motor coordination in the accelerating rota-rod test and exhibited hyperlocomotion in novel environmental and stressful conditions, when assessed by the open-field and forced-swim tests. These results indicate that the inhibitory mGluR2 plays a pivotal role in synaptic regulation of glutamatergic transmission in the neural network. PMID- 15753325 TI - Role of the renin-angiotensin system in the pathogenesis of preeclampsia. AB - Preeclampsia is a hypertensive disorder unique to pregnancy with consistent involvement of the kidney. The renin-angiotensin system (RAS) has been implicated in the pathogenesis of preeclampsia. In the gravid state, in addition to the RAS in the kidney, there is a tissue-based RAS in the uteroplacental unit. Increased renin expression observed both in human preeclampsia and in a transgenic mouse model with a human preeclampsia-like syndrome supports the concept that activation of the uteroplacental RAS, with angiotensin II entering the systemic circulation, may mediate the pathogenesis of preeclampsia. A novel disease paradigm of the two-kidney one-clip (2K-1C) Goldblatt model is presented for preeclampsia, wherein the gravid uterus is the clipped "kidney" and the two maternal kidneys represent the unclipped kidney. Validation of the 2K-1C Goldblatt model analogy requires evidence of elevated angiotensin II in the peripheral circulation before vascular maladaptation in preeclampsia. Convincing evidence of the elevation of angiotensin II in preeclampsia does not exist despite the fact that much of vascular pathogenesis appears to be due to angiotensin type I (AT(1)) receptor activation. Vascular maladaptation with increased vasomotor tone, endothelial dysfunction, and increased sensitivity to angiotensin II and norepinephrine in manifest preeclampsia may be explained on the basis of angiotensin II-mediated mechanisms. Recently, novel angiotensin II related biomolecular mechanisms have been described in preeclampsia. These include AT(1) and bradykinin B(2) receptor heterodimerization and the production of an autoantibody against AT(1). Various organ systems with a predilection for involvement in preeclampsia are each a site of a tissue-based RAS. How angiotensin II-mediated mechanisms may explain the primary clinical-pathological features of preeclampsia is described. Future investigations are proposed to more precisely define the role of activation of the uteroplacental RAS in the mechanisms underlying preeclampsia. PMID- 15753324 TI - Ficoll and dextran vs. globular proteins as probes for testing glomerular permselectivity: effects of molecular size, shape, charge, and deformability. AB - Polydisperse mixtures of dextran or Ficoll have been frequently used as molecular probes for studies of glomerular permselectivity because they are largely inert and not processed (reabsorbed) by the proximal tubules. However, dextrans are linear, flexible molecules, which apparently are hyperpermeable across the glomerular barrier. By contrast, the Ficoll molecule is almost spherical. Still, there is ample evidence that Ficoll fractional clearances (sieving coefficients) across the glomerular capillary wall (GCW) are markedly higher than those for neutral globular proteins of an equivalent in vitro Stokes-Einstein (SE) radius. Physical data, obtained by "crowding" experiments or measurements of intrinsic viscosity, suggest that the Ficoll molecule exhibits a rather open, deformable structure and thus deviates from an ideally hard sphere. This is also indicated from the relationship between (log) in vitro SE radius and (log) molecular weight (MW). Whereas globular proteins seem to behave in a way similar to hydrated hard spheres, polydisperse dextran and Ficoll exhibit in vitro SE radii that are much larger than those for compact spherical molecules of equivalent MW. For dextran, this can be partially explained by a high-molecular-size asymmetry. However, for Ficoll the explanation may be that the Ficoll molecule is more flexible (deformable) than are globular proteins. An increased compressibility of Ficoll and an increased deformability and size asymmetry for dextran may be the explanation for the fact that the permeability of the GCW is significantly higher when assessed using polysaccharides such as Ficoll or dextran compared with that obtained using globular proteins as molecular size probes. We suggest that molecular deformability, besides molecular size, shape, and charge, plays a crucial role in determining the glomerular permeability to molecules of different species. PMID- 15753326 TI - Testing the usefulness of the molecular coancestry information to assess genetic relationships in livestock using a set of Spanish sheep breeds. AB - Recent studies have proposed the use of molecular coancestry coefficients as a measure of genetic variability and as a useful tool for conservation purposes. Using simulated data, molecular coancestry has been shown to become constant very quickly after separation of populations, leading to population diversity remaining constant. However, the use of molecular coancestry information to study the genetic relationships between breeds has not yet been widely explored. Here we analyze the polymorphism of 14 microsatellites in 222 unrelated individuals belonging to seven native Spanish breeds to ascertain the usefulness of molecular coancestry-based methodologies in providing information on their genetic relationships. Average kinship distance (D(k)) and average molecular coancestry coefficients (f(ij)) were compared with well-known genetic distances, such as between-breed Reynolds' distance (D(R)), Nei's standard distance (D(s)), and shared allele distance (D(AS)). Kinship distance and f(ij) have moderate to low correlations with the other genetic distances, showing that they provide different information: both D(k) and f(ij) account for the allele frequencies in the founder population, whereas D(R), D(s), and D(AS) characterize the short-term evolution of the populations. Furthermore, D(k) and f(ij) were only moderately correlated (-0.500). The present study used field data to confirm previous research pointing out the ability of molecular coancestry coefficients to assess genetic differentiation of an ancestral origin. In this respect, molecular coancestry-based parameters may be used with classical genetic parameters to obtain information on population dynamics in livestock breeds. This study additionally presents reliable evidence on the history of these sheep breeds. PMID- 15753327 TI - Technical note: determination of alleles of the ovine PRNP gene using PCR-single strand conformational polymorphism analysis. AB - Susceptibility to scrapie in sheep is linked to variation at codons 136, 154, and 171 in the host prion protein gene (PRNP). A number of techniques are available for detecting these polymorphisms, but none allow for a rapid and accurate determination of genotype. Here we describe PCR coupled with single-strand conformational polymorphism (SSCP) analysis, which allows for the accurate identification of ovine PRNP alleles. A gene region including codons 136 to 171 was amplified by PCR, and the amplimers were then denatured and subjected to electrophoresis in a nondenaturing polyacrylamide gel. Nine unique SSCP patterns, representing nine different alleles of the ovine PRNP gene, could be resolved. A new polymorphism (I/T) at codon 142 also was detected. The profiles produced by SSCP allowed for the accurate differentiation of PRNP alleles and could be employed to genotype PRNP in sheep. PMID- 15753329 TI - Genetic parameters estimated with multitrait and linear spline-random regression models using Gelbvieh early growth data. AB - Estimates of direct and maternal genetic parameters in beef cattle were obtained with a random regression model with a linear spline function (SFM) and were compared with those obtained by a multitrait model (MTM). Weight data of 18,900 Gelbvieh calves were used, of which 100, 75, and 17% had birth (BWT), weaning (WWT), and yearling (YWT) weights, respectively. The MTM analysis was conducted with a three-trait maternal animal model. The MTM included an overall linear partial fixed regression on age at recording for WWT and YWT, and direct-maternal genetic and maternal permanent environmental effects. The SFM included the same effects as MTM, plus a direct permanent environmental effect and heterogeneous residual variance. Three knots, or breakpoints, were set to 1, 205, and 365 d. (Co)variance components in both models were estimated with a Bayesian implementation via Gibbs sampling using flat priors. Because BWT had no variability of age at recording, there was good agreement between corresponding components of variance estimated from both models. For WWT and YWT, with the exception of the sum of direct permanent environmental and residual variances, there was a general tendency for SFM estimates of variances to be lower than MTM estimates. Direct and maternal heritability estimates with SFM tended to be lower than those estimated with MTM. For example, the direct heritability for YWT was 0.59 with MTM, and 0.48 with SFM. Estimated genetic correlations for direct and maternal effects with SFM were less negative than those with MTM. For example, the direct-maternal correlation for WWT was -0.43 with MTM and -0.33 with SFM. Estimates with SFM may be superior to MTM due to better modeling of age in both fixed and random effects. PMID- 15753328 TI - Comparison of three models to estimate breeding values for percentage of loin intramuscular fat in Duroc swine. AB - Three selection models were evaluated to compare selection candidate rankings based on EBV and to evaluate subsequent effects of model-derived EBV on the selection differential and expected genetic response in the population. Data were collected from carcass- and ultrasound-derived estimates of loin i.m. fat percent (IMF) in a population of Duroc swine under selection to increase IMF. The models compared were Model 1, a two-trait animal model used in the selection experiment that included ultrasound IMF from all pigs scanned and carcass IMF from pigs slaughtered to estimate breeding values for both carcass (C1) and ultrasound IMF (U1); Model 2, a single-trait animal model that included ultrasound IMF values on all pigs scanned to estimate breeding values for ultrasound IMF (U2); and Model 3, a multiple-trait animal model including carcass IMF from slaughtered pigs and the first three principal components from a total of 10 image parameters averaged across four longitudinal ultrasound images to estimate breeding values for carcass IMF (C3). Rank correlations between breeding value estimates for U1 and C1, U1 and U2, and C1 and C3 were 0.95, 0.97, and 0.92, respectively. Other rank correlations were 0.86 or less. In the selection experiment, approximately the top 10% of boars and 50% of gilts were selected. Selection differentials for pigs in Generation 3 were greatest when ranking pigs based on C1, followed by U1, U2, and C3. In addition, selection differential and estimated response were evaluated when simulating selection of the top 1, 5, and 10% of sires and 50% of dams. Results of this analysis indicated the greatest selection differential was for selection based on C1. The greatest loss in selection differential was found for selection based on C3 when selecting the top 10 and 1% of boars and 50% of gilts. The loss in estimated response when selecting varying percentages of boars and the top 50% of gilts was greatest when selection was based on C3 (16.0 to 25.8%) and least for selection based on U1 (1.3 to 10.9%). Estimated genetic change from selection based on carcass IMF was greater than selection based on ultrasound IMF. Results show that selection based on a combination of ultrasonically predicted IMF and sib carcass IMF produced the greatest selection differentials and should lead to the greatest genetic change. PMID- 15753330 TI - Genetic analysis of carcass traits of steers adjusted to age, weight, or fat thickness slaughter endpoints. AB - Carcass measurements from 1,664 steers from the Germ Plasm Utilization project at U.S. Meat Animal Research Center were used to estimate heritabilities (h(2)) of, and genetic correlations (r(g)) among, 14 carcass traits adjusted to different endpoints (age, carcass weight, and fat thickness): HCW (kg), dressing percent (DP), adjusted fat thickness (AFT, cm), LM area (LMA, cm(2)), KPH (%), marbling score (MS), yield grade (YG), predicted percentage of retail product (PRP), retail product weight (RPW, kg), fat weight (FW, kg), bone weight (BNW, kg), actual percentage retail product (RPP), fat percent (FP), and bone percent. Fixed effects in the model included breed group, feed energy level, dam age, birth year, significant (P < 0.05) interactions, covariate for days on feed, and the appropriate covariate for endpoint nested (except age) within breed group. Random effects in the model were additive genetic effect of animal and total maternal effect of dam. Parameters were estimated by REML. For some traits, estimates of h(2) and phenotypic variance changed with different endpoints. Estimates of h(2) for HCW, DP, RPW, and BNW at constant age, weight, or fat thickness were 0.27, -, and 0.41; 0.19, 0.26, and 0.18; 0.42, 0.32, and 0.50; and 0.43, 0.32, and 0.48, respectively. Magnitude and/or sign of r(g) also changed across endpoints for 54 of the 91 trait pairs. Estimates for HCW-LMA, AFT-RPW, LMA-YG, LMA-PRP, LMA-FW, LMA-RPP, and LMA-FP at constant age, weight, or fat thickness were 0.32, -, and 0.51; -0.26, -0.77, and -; -0.71, -0.89, and -0.66; 0.68, 0.85, and 0.63; -0.16, 0.51, and 0.22; 0.47, 0.57, and 0.27; and -0.44, -0.43, and -0.18, respectively. Fat thickness was highly correlated with YG (0.86 and 0.85 for common age and weight) and PRP (-0.85 and -0.82 for common age and weight), indicating that selection for decreased fat thickness would improve YG and PRP. Carcass quality, however, would be affected negatively because of moderate r(g) (0.34 and 0.35 for common age and weight) between MS and AFT. Estimates of h(2) and phenotypic variance indicate that enough genetic variation exists to change measures of carcass merit by direct selection. For some carcass traits, however, magnitude of change would depend on effect of endpoint on h(2) and phenotypic variance. Correlated responses to selection would differ depending on endpoint. PMID- 15753331 TI - Estimates of genetic parameters for feed intake, feeding behavior, and daily gain in composite ram lambs. AB - Our objective was to estimate genetic parameters for feed intake, feeding behavior, and ADG in composite ram lambs ((1/2) Columbia, (1/4) Hampshire, (1/4) Suffolk). Data were collected from 1986 to 1997 on 1,239 ram lambs from approximately 11 to 17 wk of age at the U.S. Meat Animal Research Center near Clay Center, NE. Feeding equipment consisted of an elevated pen with an entrance chute that permitted access to the feeder by only one ram lamb at a time, with disappearance of feed measured by an electronic weighing system. Ram lambs were grouped 11 per pen from 1986 to 1989, and nine per pen from 1990 to 1997. Data were edited to exclude invalid feeding events, and approximately 80% of the data remained after edits were applied. Traits analyzed were daily feed intake (DFI), event feed intake (EFI), residual feed intake (RFI), daily feeding time (DFT), event feeding time (EFT), number of daily feeding events (DFE), and ADG. Feed intake traits of DFI and EFI had estimated heritabilities of 0.25 and 0.33, respectively, whereas estimated heritability of RFI was 0.11. Heritability estimates for feeding behavior traits, including DFT, EFT, and DFE, ranged from 0.29 to 0.36. Average daily gain had an estimated heritability of 0.26. Genetic correlations were positive between all pairs of traits, except for RFI and ADG, and that estimate was essentially zero. Phenotypic correlations were generally similar to genetic correlations. Genetic correlations were large (0.80) between DFI and ADG, intermediate between DFI and RFI (0.61) and between DFT and DFE (0.55), and low (0.17 to 0.31) for the other pairs of traits, with the exception of RFI and ADG (-0.03). Genetic correlations between behavioral traits were greater than correlations between behavioral traits and measures of feed intake or ADG; however, selection for ADG and/or feed intake would be expected to cause some changes in feeding behavior. PMID- 15753332 TI - Genetic relationship between milk score and litter weight for Targhee, Columbia, Rambouillet, and Polypay sheep. AB - This study was conducted to evaluate the relationship between milk score (MS) and litter weight at 70 d (LW) for four sheep breeds in the United States. Milk score is a subjective measure of milk production used to assess milk production of range ewes when milk yield cannot be quantitatively determined. Variance components for MS and LW were estimated for each of Targhee, Columbia, Rambouillet, and Polypay breeds. Data collected from 1990 through 2000 at the U.S. Sheep Exp. Stn. in Dubois, ID, were analyzed with an animal model using REML. There were 13,900 records of MS and LW for 5,807 ewes. Records were grouped according to parity as first, second, and greater (mature), and all records (lifetime). Estimates of heritability for MS were in the range of 0.05 to 0.18 for first, 0.01 to 0.27 for second, 0.05 to 0.10 for mature, and 0.08 to 0.13 for all lifetime parity groups. Estimates of genetic correlation between MS at first and second parities ranged from 0.74 to 1.00. Similarly, mature MS was highly correlated genetically with MS at first (0.83 to 1.00) and at second (0.60 to 1.00) parities, suggesting that additive genetic value for milking ability at maturity could be evaluated as early as at first parity. Heritability estimates for LW ranged from 0.00 to 0.18 over all breeds and parity groupings. The genetic correlation between LW at first and second parity groups ranged from 0.43 to 1.00. Estimates of genetic correlation between LW at first or second parity with mature LW were mostly high and positive, except for Targhee (-0.10) and Polypay (0.14) at first parity. Litter weight for mature ewes could be improved by selection at first or second parity. Estimates of genetic correlation at first parity between MS and LW were high (1.00) for Rambouillet and Polypay, and near zero for Columbia and Targhee. At second parity, estimates of genetic correlation between MS and LW were positive and moderate for Rambouillet and Polypay but more variable for Columbia and Targhee. Estimates of genetic correlation between MS and LW were mostly positive and may be favorable with smaller estimates of standard errors using all lifetime records rather than first or second parity records. Although estimates are variable, the average of the estimates of the genetic correlation suggests that LW can be improved by selecting ewes for favorable MS. PMID- 15753333 TI - Genetic evaluation of the ratio of calf weaning weight to cow weight. AB - The phenotypic ratio of a calf's weaning weight to its dam's weight is thought to be an indicator of efficiency of the cow. Thus, the objectives of this research were to 1) estimate genetic parameters for the ratio of 200-d calf weight to mature-equivalent cow weight at weaning, its components, and other growth traits; and 2) evaluate responses to selection based on the ratio. Phenotypes evaluated were the ratio (100 kg/ kg; n = 4,184), birth weight (kg; n = 5,083), 200-d weight (kg; n = 4,902), 365-d weight (kg; n = 4,626), and mature-equivalent cow weight at weaning (kg; n = 4,375). In 1989, a randomly selected and mated control line and a line selected for greater values of the ratio were established. Average generation intervals were 3.39 +/- 0.05 and 3.90 +/- 0.08 yr in the ratio selected line and control line, respectively. The ratio selection line (n = 895) accumulated approximately 4.7 SD more selection differential than the control line (n = 912) over 2.5 generations. Data were analyzed with a multiple-trait Gibbs sampler for animal models to make Bayesian inferences. Heritability estimates (posterior mean +/- SD) for direct effects were 0.20 +/- 0.03, 0.46 +/- 0.04, 0.48 +/- 0.03, 0.58 +/- 0.04, and 0.76 +/- 0.02 for ratio, birth weight, 200-d weight, 365-d weight, and cow weight, respectively. Estimates for heritability of maternal effects were 0.58 +/- 0.05, 0.10 +/- 0.02, 0.13 +/- 0.02, and 0.10 +/- 0.02 for ratio, birth weight, 200-d weight, 365-d weight, respectively. Significant response to selection was limited to maternal effects: 1.32 +/- 0.38 ratio units per generation. As the ratio was a trait of the calf, estimated maternal genetic effects on the ratio contained both genetic effects due to dams that environmentally affected progeny performance and direct effects on the reciprocal of cow weight. In the control line, genetic trends in direct and maternal 200-d weight were -1.28 +/- 0.91 and 0.62 +/- 0.92 kg/generation, respectively, and the genetic trend in direct effects on cow weight was -5.72 +/- 2.80 kg/ generation. In the selection line, genetic trends in direct and maternal 200-d weight were 1.43 +/- 0.79 and 2.90 +/- 0.80 kg/generation and the genetic trend in cow weight was -2.79 +/- 2.43 kg/generation. Significant correlated responses were observed in direct effects on birth weight and maternal effects on 365-d weight. Results contraindicate use of the ratio of calf weaning weight to cow weight as a selection criterion. PMID- 15753334 TI - Effect of testosterone on insulin-like growth factor-I, androgen receptor, and myostatin gene expression in splenius and semitendinosus muscles in sheep. AB - Testosterone is known to act differentially on skeletal muscle from different regions. Two genes likely to mediate the testosterone effect are IGF-I, an important growth regulator acting in an autocrine and paracrine way, and androgen receptor (AR), as receptor density could account for differential muscle growth. Another muscle-specific gene that may play a role in differential muscle growth is myostatin, a member of the transforming growth factor-beta superfamily, shown to be a negative regulator of skeletal muscle mass. The objective of this study was to quantify and compare the expression of these three genes in two different skeletal muscles in sheep. East Friesian x Dorset-sired ram lambs from Dorset ewes were used in a 2 x 4 factorial experiment. Eighteen sets of twins were assigned to four age groups corresponding to 77, 105, 133, and 161 d of age, and one individual from each set was castrated at birth. Total RNA was extracted from samples of splenius (SP) and semitendinosus muscles collected at the time of slaughter. Insulin-like growth factor-I mRNA was measured using competitive reverse-transcription PCR. Androgen receptor and myostatin mRNA were measured by ribonuclease protection assay with standard curves. Weight of SP was greater than semitendinosus in rams compared with wethers at 105, 133, and 161 d (P = 0.05, P = 0.04, and P = 0.02, respectively). The difference in IGF-I mRNA levels between the two muscles was greater in rams than in wethers at 133 (P = 0.001) and 161 d (P = 0.014), and the difference in AR mRNA levels was greater in rams than in wethers at 105, 133, and 161 d (P = 0.002, P < 0.001, and P < 0.001, respectively), with greater abundance in the SP. No difference was found in myostatin mRNA level between the two muscles in rams and wethers at any age. These results suggest that locally produced IGF-I and the regulation of AR expression are important for sexually dimorphic muscle growth patterns. PMID- 15753336 TI - Effects of recombinant bovine somatotropin on growth and abundance of mRNA for IGF-I and IGF-II in channel catfish (Ictalurus punctatus). AB - Research was conducted to examine growth rates, circulating concentrations of IGF I, and mRNA abundance levels of IGF-I and IGF-II in channel catfish (Ictalurus punctatus) given recombinant bovine ST (rbST; Posilac, Monsanto Co., St. Louis MO). In the first study, juvenile catfish (5.5 +/- 0.5 g) were randomly assigned to one of three treatments: 1) sham-injected control (one needle puncture per week); 2) rbST (30 microg x g BW(-1) x wk(-1); Posilac); and 3) nonhandled control (control). At the end of the 6-wk study, the fish were weighed, measured for length, and G:F was determined. Compared with sham and control treatments, rbST-treated fish had 48% greater final BW, 14% greater total length, and 52% greater G:F (P < 0.001). In the second study, juvenile catfish (41.1 +/- 1.5 g) were assigned randomly to one of two treatments: 1) sham or 2) rbST. Eight fish per treatment were sampled on d 0, 1, 2, 7, 14, and 21 for blood, muscle, and liver. Relative expression of IGF-I and IGF-II mRNA was determined by real-time PCR and plasma concentrations of IGF-I were measured using a validated fluoroimmunoassay. Circulating concentrations of IGF-I were increased (37.9 +/- 5.5 vs. 22.0 +/- 6.6 ng/mL; P < 0.05) in rbST-injected fish compared with sham injected controls by d 14. Liver IGF-I and IGF-II mRNA was increased 4.3-and 14.4 fold, respectively, by d 1 in rbST-injected fish compared with controls (P < 0.05); however, abundance of liver IGF-I and IGF-II mRNA did not differ from controls on d 0, 2, 7, 14, and 21. Abundance of muscle IGF-I and IGF-II mRNA did not differ in rbST-injected fish compared with controls throughout the study. Results of the first study demonstrated that rbST improves growth performance of channel catfish. Results of the second study showed that the growth-promoting effects of rbST were not mediated by the expression of IGF-I or IGF-II mRNA in the muscle. Instead, the results suggest that rbST promotes growth by stimulating plasma IGF-I release, possibly through its direct effect on the liver or on local tissues to synthesize IGF-I. The changes in mRNA abundance and plasma concentrations of IGF-I support the role of IGF-I in growth regulation of channel catfish. PMID- 15753335 TI - Growth hormone at breeding modifies conceptus development and postnatal growth in sheep. AB - Experiments were performed to determine the effects of components of the GH-IGF axis on conceptus development and postnatal growth in sheep. In Exp. 1, ewes received one of the following treatments: 1) sustained release GH at breeding, 2) sustained release GH at breeding and estradiol-17beta at d 5 and 6, 3) only estradiol-17beta at d 5 and 6, or 4) no treatment. Uteri were flushed on d 7, and flushings were analyzed for content of IGF-I. A single injection of sustained release bovine GH at breeding increased IGF-I content in uterine luminal flushings compared with control ewes (P < 0.05). Treatment with estradiol-17beta on d 5 and 6 after breeding did not alter IGF-I content compared with control ewes, and it blocked the effect of GH on uterine luminal IGF-I content. In Exp. 2, sustained release GH or no treatment was administered at breeding, and gravid uteri were collected at d 25, 80, or 140 of gestation. On d 80, GH-treated ewes had smaller chorioallantoic weights (P < 0.05) and tended to have more efficient placentae (fetal weight/total placental weight; P = 0.052), with a higher percentage of placental weight as cotyledons (P = 0.068) compared with control ewes. In Exp. 3, ewes were treated with or without sustained release GH at progesterone withdrawal. Lambs from GH-treated ewes were heavier at birth (P < 0.05). Lambs from GH-treated ewes reared as singles, but not lambs reared as multiples, were heavier at 30, 60 (P < 0.05), and 75 d (P = 0.075) of age than lambs from control ewes. In conclusion, ewes treated with sustained-release GH at breeding developed smaller, more efficient placentas, and had larger lambs at birth. PMID- 15753337 TI - Exogenous prolactin stimulates mammary development and alters expression of prolactin-related genes in prepubertal gilts. AB - The goal of this project was to determine whether recombinant porcine (rp) prolactin (PRL) can enhance mammary development when given to pre-pubertal gilts and/or modify the expression of PRL-related genes. Crossbred gilts were injected s.c. twice daily with saline (CTRL; n = 13), 2 mg of rpPRL (4PRL; n = 13), or 4 mg of rpPRL (8PRL; n = 13) in a 2.0-mL volume for a period of 29 d, starting at 75.1 +/- 0.5 kg BW. Jugular blood samples were collected before the first injection, as well as 14 and 28 d later, and were assayed for PRL, IGF-I, and leptin. Gilts were slaughtered on d 29 of treatment, and mammary glands were collected for dissection of parenchymal and extraparenchymal tissues, and for determination of parenchymal DNA, DM, protein, and fat contents. Levels of mRNA for PRL, PRL receptor (PRL-R), and signal transducers and activators of transcription (STAT5A and STAT5B) were determined via real-time PCR in the mammary parenchyma, as well as levels for PRL and PRL-R in the pituitaries. Treatments did not alter plasma (P = 0.48) IGF-I. Serum concentrations of PRL at slaughter were greater (P < 0.01) in both 4PRL and 8PRL compared with CTRL, whereas at mid-treatment, they were greater (P < 0.05) only in 8PRL gilts. Parenchymal tissue weight and parenchymal DNA concentrations increased with exogenous rpPRL (P < 0.001). The percentage of protein in parenchyma increased (P < 0.001), whereas that of DM (P < 0.001), fat (P < 0.001), and the protein:DNA ratio (P < 0.05) decreased with exogenous rpPRL. Treatment differences were always observed between the 4 mg dose and CTRL, and no further differences were noted when the dose was increased to 8 mg daily. Expression levels of PRL, but not PRL-R, were decreased (P < 0.05) in anterior pituitary glands and mammary glands of treated gilts. The mRNA levels of STAT5A and STAT5B increased (P < 0.05) with exogenous rpPRL. It is evident from these data that rpPRL can stimulate mammogenesis in prepubertal gilts through hyperplasia and increased expression of PRL-related genes. PMID- 15753338 TI - Response to lysine in a wheat gluten diet in adult minipigs after short-and long term dietary adaptation as assessed with an indicator amino acid oxidation and balance technique. AB - An experiment was conducted to examine the response to wheat gluten (WG)-based diets at two lysine levels in adult minipigs (23 kg BW) using the indicator AA oxidation (IAAO) approach and N balance. Twenty minipigs (n = five per group), fitted with reentrant ileoileal cannulas allowing collection of ileal effluents, were fed restrictively two WG-based diets (WG and WG + Lys; 2.7 and 6.6 g of lysine/kg, respectively) for adaptation periods of 10 and 100 d. On d 7 and 9, for pigs fed the diets for 10 d, and on d 97 and 99, for pigs fed the diets for 100 d, primed i.v. fasted/fed tracer protocols with [(13)C]bicarbonate, and [(13)C]leucine were performed. With the WG diet, [(13)C]bicarbonate recoveries (%) were lower irrespective of the adaptation period, and higher during the fed period (fasted: WG + Lys = 82.5, and WG = 69.1; fed: WG + Lys = 90.6, and WG = 85.9; P < 0.05). Leucine oxidation rate was higher with the lower lysine intake (WG = 194.6 vs. 109.5 mg/[kg BW x d]; P < 0.05). Wheat gluten feeding resulted in a negative leucine balance independent of the adaptation period (WG = -29.1, and WG + Lys = 48.2 mg/[kg BW x d]; P < 0.05). In contrast with the IAAO method, N balance did not differ between the two lysine intakes, possibly because of an underestimation of N losses. The finding of a lower (13)C bicarbonate recovery with the lower dietary lysine intake suggests that caution should be taken in using a single recovery factor for all AA oxidation studies. PMID- 15753339 TI - Comparing efficiency of metabolizable energy utilization by rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) using factorial and multivariate approaches. AB - A study was conducted to compare utilization of ME for growth vs. maintenance in rainbow trout and Atlantic salmon. Fish were hand-fed to satiation one of four isoenergetic diets (DE = 20 MJ/kg, as-fed basis) with different digestible protein (DP) to DE ratios (24, 22, 20, and 18 g/MJ). Intake of ME (kJ/d), energy deposited as protein (PD, kJ/d), and energy deposited as lipid (LD, kJ/d) were determined by a comparative slaughter technique. Data were analyzed by a factorial approach or by multivariate analysis of PD and LD on ME. Maintenance energy requirements (ME(m)) and efficiency of ME utilization for PD (k(p)) and LD (k(f)) were estimated with both approaches. For the multivariate analysis, an additional parameter, the fraction of ME intake above maintenance used for PD (X) was defined as linear function of BW, with slope (d) and intercept (c) estimated simultaneously with the above parameters. Estimates were highly dependent on the approach and assumptions used. The ME(m) and k(p) values were higher and less accurate with the factorial approach than with multivariate analysis. The factorial approach estimated unrealistic k(f) values (k(f) > 1). With the multivariate analysis, ME(m) did not differ between species (20 kJ x d(-1) x kg( 0.8)). On the other hand, k(p) was significantly higher (e.g., 0.52 +/- 0.06 vs. 0.43 +/- 0.06; P < 0.05) for salmon than for trout and independent of diet, but k(f) was 0.81 (+/-0.13) regardless of species or diet. The ME intake above ME(m) used for PD (c) was higher in salmon than trout (57 vs. 55%; P < 0.05). The change in partitioning of ME for PD due to the change in BW was negative for trout (d = -0.18), but positive for salmon (d = 0.16). The d values agreed well with the increase of LD:PD ratio with BW for trout and the decrease of LD:PD with BW for salmon, which may have been related to the maturation status of this fish and the associated loss of body lipid observed by maturing salmon. In conclusion, ME(m) and cost of LD were similar for rainbow trout and Atlantic salmon, but the cost of PD was lower for salmon than for trout. PMID- 15753340 TI - Comparison of dietary selenium fed to grower-finisher pigs from various regions of the United States on resulting tissue Se and loin mineral concentrations. AB - A study was conducted to evaluate the mineral content of pork tissue with particular emphasis on Se between various states (regions) having different diet (grain) indigenous Se concentrations. The study involved 19 states in the north, central, and southern regions of the United States, with committee members of NCR 42 and S-1012 (formerly S-288). A total of 62 pigs were used, with collaborators sending 100-g samples each of loin, heart, and liver, and a 3- to 4-g sample of hair (collected along the topline) from two to five market-weight pigs to a common laboratory for analysis. Diets at each station were formulated with locally purchased soybean meal and grain that was either grown or normally fed to pigs within their state. Tissues were analyzed for Se, but only the loin was analyzed for the macro- and micromineral elements. Correlation of dietary minerals to the tissue element was determined. The results demonstrated differences in tissue Se among states (P < 0.01), with high correlations of dietary Se to loin (r = 0.84; P < 0.01), heart (r = 0.84; P < 0.01), liver (r = 0.83; P < 0.01), and hair Se (r = 0.90; P < 0.01) concentrations. The correlation of hair Se to the Se concentration of loin, heart, and liver tissues was high (r > 0.90; P < 0.01). States in the west-central region of the United States and west of the Mississippi river had higher dietary Se and tissue Se concentrations than states in the eastern section of the Corn Belt, east of the Mississippi river, and along the East Coast. Generally, states did not differ greatly in their loin macro- and micromineral concentrations. The simple correlation of dietary minerals to their corresponding loin mineral concentration was generally non-significant, but most macrominerals had decreasing mineral concentrations when the dietary mineral level was higher. These results indicate that regional differences in tissue Se were influenced more by the indigenous Se content of the diet (grain) fed to the pigs than from sodium selenite. PMID- 15753341 TI - Effects of chromium propionate on growth, carcass traits, and pork quality of growing-finishing pigs. AB - An experiment was conducted to determine the effect of dietary Cr propionate (CrProp) on growth, carcass traits, and pork quality of crossbred finishing gilts. Dietary treatments were 0 or 200 ppb Cr (as CrProp; as-fed basis), and each treatment was replicated four times with five gilts per replicate pen. Gilts were fed diets containing 0.82% lysine from 73 to 80 kg BW and 0.64% lysine from 80 to 115 kg BW. At the end of the trial, carcass and pork quality data were collected from four gilts per replicate. Average daily gain, ADFI, and G:F were not affected (P = 0.76 to 0.96) by CrProp. Before delivery at the abattoir, shrink loss was determined after an 18-h fast (fasting shrink) and after hauling (shipping shrink) pigs for 2.66 h (209.2 km). Fasting, shipping, and overall shrink were not affected (P = 0.14 to 0.39) by CrProp. Carcass length was increased (P = 0.03) in pigs fed CrProp. Loin muscle area, 10th-rib backfat thickness, average backfat thickness, dressing percent, muscle score, fat-free lean, and percent lean were not affected (P = 0.18 to 0.95) by CrProp. Twenty four-hour loin pH was increased (P = 0.10) in pigs fed CrProp, but 45-min loin and ham pH and 24-h ham pH were not affected (P = 0.39 to 0.83) by CrProp. Subjective (color, marbling, firmness, and wetness) and objective (Commission Internationale de l'Eclairage L*, a*, b*) assessments of the loin muscle (at the 10th-rib interface) were not affected (P = 0.62 to 0.99) by CrProp. Forty-eight hour drip (P = 0.10) and 21-d purge loss (P = 0.01) were decreased in pigs fed CrProp, but cook and total loss (drip + cook loss) and shear force were not affected (P = 0.35 to 0.53) by CrProp. Plasma cortisol, glucose, and lactate concentrations were not affected (P = 0.28 to 0.97) by CrProp after transportation or during exsanguination. These data indicate that CrProp may improve some aspects of pork quality (loin pH, drip and purge loss) but not growth performance or carcass traits. PMID- 15753342 TI - Level of supplemental protein does not influence the ruminally undegradable protein value. AB - Two experiments were conducted to determine whether elevating the percentage of ruminally undegradable protein (RUP) in the diet would influence the RUP value of the protein feedstuff. A single-effluent, continuous-culture study was designed to test the effect of RUP inclusion rate in the diet on ruminal degradability of the protein. Treatments consisted (DM basis) of a control diet with no supplemental protein, control + 2.5% bloodmeal (BM-L), control + 5% bloodmeal (BM H), control + 4.45% soybean meal (SBM-L), and control + 8.89% soybean meal (SBM H). Proteolytic activity and total VFA concentration were not affected (P = 0.73 and P = 0.13) by treatment. Within protein source, dietary RUP value was not affected (P = 0.94) by level of inclusion. When corrected for control diet RUP flow, the RUP value of the blood meal (BM) protein was higher (P = 0.01) than soybean meal (SBM); however, level of supplementation did not affect (P = 0.07) the RUP value of BM or SBM. In Exp. 2, 32 British x Continental crossbred steers (276 +/- 26.3 kg) were fed for 72 d to examine the effects of balancing the AA:energy ratio, using BM as a RUP source, on ADG, G:F, and lean tissue deposition. Diets were formulated to provide increasing levels of arginine, while ruminally degradable protein and energy were held constant. Four dietary treatments provided 0.5, 1, 1.5, and 2x the required amount of arginine, whereas the control diet had no BM included. Daily DMI averaged 7.6 kg/steer and did not differ (P = 0.71) among treatments. Steers gained an average of 1.9 kg/d and average G:F was 0.260, with no differences (P = 0.60 and P = 0.97, respectively) among treatments. There was no difference (P = 0.48) in the change in 12th-rib fat depth during the study; however, change in LM area was affected quadratically as the level of BM increased in the diet, with the greatest increase in LM area occurring in steers fed the 1x and 1.5x required arginine treatments. Balancing the AA:energy ratio did not affect G:F, DMI, or ADG; however, it increased deposition of lean in the LM quadratically. Level of dietary inclusion of BM as an RUP source does not affect its RUP value or efficacy of providing postruminal AA in growing steers. PMID- 15753343 TI - High mineral and vitamin E intake by pregnant ewes lowers colostral immunoglobulin G absorption by the lamb. AB - A 2 x 2 factorial arrangement of treatments with 78 mature ewes was used to evaluate the effects of supplementing the pregnant ewe's diet with high levels of minerals and vitamin E on immunoglobulin G (IgG) absorption by the lamb and whether any altered efficacy of IgG absorption was due to the colostrum or to the lamb. The ewes were estrus-synchronized in October and housed in wk 10 of gestation. In the final 7 wk of gestation, a grass silage-based diet, offered ad libitum, was supplemented with 500 g of a 19% CP concentrate, and from 1 wk later until lambing, half the ewes was offered 48 g of a mineral/vitamin supplement containing 6.5 g of Ca, 4.9 g of P, 5.9 g of Mg, 4.0 g of Na, 790 mg of Zn, 3.5 mg of Se, 40 mg of I, 200 mg of Mn, 20 mg of Co, and 40 IU of vitamin E. At birth, the lambs were allocated to one of four treatments in a 2 x 2 factorial arrangement, with lamb origin and colostrum origin as the two factors. The lambs born to ewes not offered the mineral supplement were fed colostrum obtained from their own dams or from ewes in the mineral-supplemented treatment, whereas lambs born to ewes given supplemental minerals were fed colostrum obtained either from their dams or from ewes in the control treatment. The ewes were milked at 1, 10, and 18 h postpartum and the lambs were fed using a stomach tube. A 5-mL blood sample was taken from each lamb at 24 h postpartum for IgG analysis. The level of fecal adhesion to the upper tail/rump area of the lamb was subjectively scored at 72 h postpartum. There was no difference in gestation length, lamb birth weight, colostrum yield, or IgG production (P = 0.16 to 0.82). When ewes were fed supplemental minerals, the serum IgG content of the progeny was lower than in their control counterparts (6.8 vs. 16.1 g/L; P < 0.001), regardless of whether the lamb received colostrum from ewes with or without access to supplementary minerals. The difference in serum IgG concentrations at 24 h postpartum was a direct reflection of a compromised efficiency in IgG absorption. The progeny of ewes with access to minerals had higher (P < 0.05) levels of fecal adhesion, which was not related to the origin of the colostrum, indicating altered digestive function in these lambs. We conclude, using the sheep as a model, that high mineral intakes in late pregnancy not only lower serum IgG concentrations in the lamb, but also that high mineral intakes result in the neonate being preprogrammed at birth so that it is born with a compromised ability to absorb colostral IgG. PMID- 15753344 TI - Using blood urea nitrogen to predict nitrogen excretion and efficiency of nitrogen utilization in cattle, sheep, goats, horses, pigs, and rats. AB - The objectives of this study were to evaluate the potential for using blood urea N concentration to predict urinary N excretion rate, and to develop a mathematical model to estimate important variables of N utilization for several different species of farm animals and for rats. Treatment means (n = 251) from 41 research publications were used to develop mathematical relationships. There was a strong linear relationship between blood urea N concentration (mg/100 mL) and rate of N excretion (g x d(-1) x kg BW(-1)) for all animal species investigated. The N clearance rate of the kidney (L of blood cleared of urea x d(-1) x kg BW( 1)) was greater for pigs and rats than for herbivores (cattle, sheep, goats, horses). A model was developed to estimate parameters of N utilization. Driving variables for the model included blood urea N concentration (mg/100 mL), BW (kg), milk production rate (kg/d), and ADG (kg/d), and response variables included urinary N excretion rate (g/d), fecal N excretion rate (g/d), rate of N intake (g/d), and N utilization efficiency (N in milk and gain per unit of N intake). Prediction errors varied widely depending on the variable and species of animal, with most of the variation attributed to study differences. Blood urea N concentration (mg/100 mL) can be used to predict relative differences in urinary N excretion rate (g/d) for animals of a similar type and stage of production within a study, but is less reliable across animal types or studies. Blood urea N concentration (mg/100 mL) can be further integrated with estimates of N digestibility (g/g) and N retention (g/d) to predict fecal N (g/d), N intake (g/d), and N utilization efficiency (grams of N in milk and meat per gram of N intake). Target values of blood urea N concentration (mg/100 mL) can be backcalculated from required dietary N (g/d) and expected protein digestibility. Blood urea N can be used in various animal species to quantify N utilization and excretion rates. PMID- 15753345 TI - Effects of marbling and shear force on consumers' willingness to pay for beef strip loin steaks. AB - Experimental economic procedures were used to measure the effects of changes in marbling score and Warner-Bratzler shear force (WBSF) value on consumer purchasing behavior and willingness to pay for beef strip loin steaks (n = 541). Consumers were more likely to bid on a steak during the experimental auction if the steak had a high marbling score or low WBSF value. Averaging across all consumers in the study (n = 489), the predicted odds that consumers would submit a nonzero bid were favorable for steaks with a marbling score greater than Modest(50) or a WBSF value less than 3.9 kg. Bid prices for steaks were analyzed with respect to changes in steak marbling score, WBSF value, quality grade marketing category classification (Select, Low Choice, Premium Choice, and Prime), and WBSF marketing category classification (very tender, 5.40 kg). The percentage of bids that were zero was highest (P < 0.05) for Select steaks, intermediate (P < 0.05) for Low Choice steaks, and lower (P <0.05) for Premium Choice or Prime steaks. Steaks in the very tender category had the lowest (P < 0.05) percentage of zero bids, and steaks in the slightly tough and very tough categories had the highest (P < 0.05) percentage of zero bids submitted from "buyers" in the auction. Premium Choice and Prime steaks were valued higher (P < 0.05) than Select steaks by consumers. On average, Premium Choice steaks received a 0.89 dollars/kg premium, and Prime steaks received a 2.47 dollars/kg premium over the mean bid price for Select steaks. Predicted mean bid prices for steaks decreased by 1.02 dollars/kg for each 1 kg increase in WBSF value. On average, steaks in the very tender marketing category received a higher (P < 0.05) bid price than the bid price for steaks in the slightly tender, slightly tough, and very tough categories (+0.83 dollars/kg, +2.09 dollars/kg, and +2.55 dollars/kg, respectively). Mean bid prices for steaks from the slightly tough and very tough categories did not differ (P = 0.184). Compared with the mean bid price for steaks in the slightly tender category, steaks from the slightly tough and very tough categories were discounted (P < 0.05) by 1.26 dollars/kg and 1.72 dollars/kg, respectively. Results suggest that marbling score and WBSF influence both the probability that consumers will purchase and the price they are willing to pay for strip loin steaks. PMID- 15753346 TI - Preslaughter handling effects on pork quality and glycolytic potential in two muscles differing in fiber type composition. AB - The objective of the present experiment was to investigate the effects of transportation, lairage, and preslaughter stressor treatment on glycolytic potential and pork quality of the glycolytic longissimus and the oxidative supraspinatus (SSP) or serratus ventralis (SV) muscles. In a 2 x 2 x 2 factorial design, 384 pigs were assigned randomly either to short (50 min) and smooth or long (3 h) and rough transport, long (3 h) or short (< 45 min) lairage, and minimal or high preslaughter stress. Muscle samples were taken from the LM at 135 min and from the SSP at 160 min postmortem for determination of the glycolytic potential and rate of glycolysis. At 23 h postmortem, pork quality was assessed in the LM and the SV. Effects of transport and lairage conditions were similar in both muscle types. Long transport increased (P < 0.01) the glycolytic potential and muscle lactate concentrations compared with short transport. Both long transportation and short lairage decreased (P < 0.01) redness (a* values) and yellowness (b* values) of the LM and SV. In combination with short lairage, long transport decreased (P < 0.05) pork lightness (lower L* values), and electrical conductivity was increased (P < 0.05) after long transport. Several interactions between stress level and muscle type (P < 0.001) were observed. High preslaughter stress decreased (P < 0.001) muscle glycogen in both the LM and SSP, but this decrease was greater in the LM. Lactate concentrations were increased (P < 0.001) only in the LM by high preslaughter stress. Increases in ultimate pH (P < 0.001) and decreases in a* values (P < 0.01) were greatest in the SV, whereas increases in electrical conductivity (P < 0.001) were greatest in the LM. The lack of interactions among transportation, lairage, and muscle type was attributed to the relatively minor differences in stress among treatments. It was concluded that, in glycolytic muscle types such as the LM, the high physical and psychological stress levels associated with stress in the immediate preslaughter period have a greater effect on the water-holding capacity of the meat and may promote PSE development. Conversely, oxidative muscle types tend to have higher ultimate pH values and produce DFD pork in response to intense physical activity and/or high psychological stress levels preslaughter. PMID- 15753347 TI - Effects of grouping unfamiliar cohorts, high ambient temperature and stocking density on live performance of growing pigs. AB - Ninety-six crossbred intact male pigs (34.5 +/- 3.5 kg BW) were allocated by weight and vocalization score to a 2 x 2 x 2 dynamic experimental design including two stocking densities (1 or 2 m(2)/pig), two temperatures (22 degrees C and 30 degrees C), and two short groupings of unfamiliar cohorts (six pigs as one pig per group, and six pigs per group). The study was conducted over 8 wk, and live weight gain (WTG) and feed intake (FI; as-fed basis) were measured weekly. During the first week, pigs were housed in individual pens from four independent rooms. To group pigs, pen partitions were removed. Pigs were grouped in Rooms 2 and 3 from wk 2 to 4, and in Rooms 1 and 4 during wk 7. Temperature was increased from 22 degrees C to 30 degrees C in Rooms 1 and 2 during wk 4 and 7. Pen partitions were replaced in Rooms 2 and 3 at the end of wk 4 and in Rooms 1 and 4 at the end of wk 7 to return pigs to their individual pens. Grouping pigs decreased FI during wk 3 (15.08 +/- 0.43 vs. 14.03 +/- 0.41 kg P < 0.10), and during wk 7 (17.42 +/- 0.46 vs. 14.24 +/- 0.41 kg; P < 0.01). In addition, grouping had a negative effect (P < 0.001) on WTG at wk 3 (7.38 +/- 0.28 vs. 5.71 +/- 0.28 kg) and at wk 7 (6.70 +/- 0.26 vs. 2.99 +/- 0.26 kg). For grouped pigs, raising the temperature decreased (P < 0.01) WTG (7.49 +/- 0.29 vs. 6.41 +/- 0.29 kg during wk 4, and 3.37 +/- 0.38 vs. 2.62 +/- 0.38 kg during wk 7). Mean FI was decreased (P < 0.01) with the 30 degrees C treatment during wk 7 only (15.49 +/- 0.33 kg at 22 degrees C compared with 12.99 +/- 0.33 kg at 30 degrees C). Compensatory feed intake was evident after the treatments had ceased at wk 6, whereby previously heat-treated grouped pigs had a higher FI (17.97 +/- 0.45 kg) than the animals individually housed at 22 degrees C (12.99 +/- 0.33 kg). Stocking density effects were noted after the grouping and high temperature treatments had ceased. For instance, during wk 5, low-density-housed pigs grew faster (P < 0.001) than their high-density counterparts (9.04 +/- 0.38 vs. 7.49 +/- 0.29 kg). In conclusion, under the conditions of this study, the grouping of unfamiliar cohorts and high ambient temperature treatments had a detrimental effect on pig performance, and these effects were reversible. PMID- 15753348 TI - Nutritionally altering weight gain patterns of pregnant heifers and young cows changes the time that feed resources are offered without any differences in production. AB - We hypothesized that feed resources could be deferred to a later time in the production cycle without a decrease in fertility or weight of calf produced in heifers and young cows. One-hundred and thirty-one MARC III (four breed composite: (1/4) Hereford, (1/4) Angus, (1/4) Red Poll, and (1/4) Pinzgauer) heifers were divided into three treatments: M-M-M-M (n = 46), L-H-M-M (n = 41), and L-L-L-H (n = 44). The experiment consisted of four feeding periods. Period 1 was 94 to 186 d of gestation, and heifers were fed a moderate (M) or low (L) level of feed. Period 2 was 187 d of gestation to parturition, and heifers were fed moderate, high (H), or low levels of feed. Period 3 was from parturition through 27 d of lactation, and heifers were fed moderate or low levels of feed. Period 4 was from 28 d to approximately 63 d of lactation, and heifers were fed moderate or high levels of feed. Females remained within treatments through their first parity (heifers) and second parity (cows). Feed intake of L-H-M-M and M-M-M M treatments did not differ from each other either as heifers (P = 0.23) or as second-parity cows (P > 0.59). The L-L-L-H heifers ate less feed than L-H-M-M and M-M-M-M heifers (P < 0.001), and second-parity L-L-L-H cows ate less feed than second-parity L-H-M-M and M-M-M-M cows (P < 0.002). In the first parity, treatments did not differ in the percentage of calves weaned (P = 0.11), weight of calf weaned (P = 0.50), or percentage of cows diagnosed pregnant (P = 0.29) with a second calf. In the second parity, treatments did not differ in the percentage of calves weaned (P = 0.77), weight of calf weaned (P = 0.63), or percentage of cows expressing a corpus luteum at the start of breeding for their third calf (P = 0.21). Our findings suggest that timing nutrient availability to heifers and primiparous cows can be used to change the time that feed resources are used. PMID- 15753349 TI - The effect of a leptin single nucleotide polymorphism on quality grade, yield grade, and carcass weight of beef cattle. AB - Feedlot producers could optimize the value of cattle in a given market grid if they were able to improve the uniformity of the body composition between cattle among loads. Allelic variation due to a single nucleotide transition (cytosine [C] to thymine [T] transition that results in a Arg25Cys) has been demonstrated to be associated with higher leptin mRNA levels in adipose tissue and increased fat deposition in mature beef, but the effect on economically important carcass traits has not been investigated in either market-ready steers or heifers. Therefore, the objective of this study was to determine the effects of a leptin SNP on the quality grade (QG), yield grade (YG), and weight of beef carcasses. A slaughter trial was conducted using 1,435 crossbred finished heifers and 142 crossbred finished steers as they entered the slaughter facility. Canada QG tended (main effect of genotype P = 0.16, but P < 0.01 for both CC vs. TT and CT vs. TT) to be affected by leptin genotype. Specifically, 7.6 and 7.1% more TT carcasses graded Canada AAA or higher than the CT and CC carcasses, respectively, which supports the suggestion that the leptin SNP is associated with carcass fat. The proportion of carcasses grading Canada YG 1, 2, or 3 was affected (P < 0.01, P = 0.05, and P = 0.02 for YG 1, 2, and 3) by leptin genotype. The proportion of TT carcasses of Canada YG 1 was 12.5 and 15.1% lower than that of CT and CC carcasses, respectively, indicating that rearing animals under the same management and feeding system may result in greater carcass fat and a lower probability of the proportion of carcasses grading YG 1 within certain genotypes. The carcass weights of animals with the CC genotype tended (P = 0.07) to be higher than those of the TT genotype (365.5 vs. 362.3 kg). No significant difference was observed between the TT and CT genotypes in carcass weight. The observed associations between leptin genotype and carcass characteristics may represent an opportunity to genetically identify animals that are most likely to reach specific marketing groups. PMID- 15753350 TI - The effect of early weaning on feedlot performance and measures of stress in beef calves. AB - Forty crossbred steers (Brahman x English) were categorized into two groups: 1) early weaned (EW; n = 20); and 2) normal weaned (NW; n = 20). Calves were 89 and 300 d of age at the time of EW and NW, respectively; SEM = 4.4. Early-weaned calves were kept on-site (University of Florida, Ona), provided supplement (1% of BW), and grazed on annual and perennial pastures until NW. At the time of normal weaning, all calves were loaded on a commercial livestock trailer and transported to the North Carolina State University Research Feedlot in Butner (approximately 1,200 km). Upon arrival, calves were stratified by BW and randomly allotted to four pens per weaning age treatment. Individual calf BW and blood samples were collected at the time of normal weaning, on arrival at the feedlot (d 1; 24 h following weaning), and on d 3, 7, 14, 21, and 28 of the receiving period. Individual BW was collected at the start and end of the growing and finishing periods, and feed intake by pen was measured daily. As an estimate of stress during the receiving period, plasma was collected and analyzed for the acute phase proteins, haptoglobin and ceruloplasmin. Early-weaned calves were lighter (P = 0.03) at normal weaning than NW calves (221 vs. 269 kg; SEM = 10.6). By d 28, EW calves tended (P = 0.12) to be lighter than NW calves (242 vs. 282 kg, respectively). Gain:feed was improved for EW compared with NW calves during both the receiving (G:F = 0.157 vs. 0.081) and growing (0.159 vs. 0.136) periods. There tended (P < 0.10) to be weaning age x day interactions for each acute-phase protein. Ceruloplasmin concentrations increased in NW, but not EW calves, and peaked on d 7 (27.6 and 34.2 mg/100 mL for EW and NW calves, respectively; P < 0.05). Haptoglobin concentrations increased in both groups and were greatest (P < 0.05) in NW calves on d 3 (7.63 vs. 14.86 mg of haptoglobin/hemoglobin complexing/100 mL). No differences in ADG or G:F were detected during the finishing phase; however, overall G:F was improved (P = 0.03) for EW vs. NW calves (0.155 vs. 0.136). Carcass measures, including backfat thickness, USDA yield grade, marbling score, and LM area, did not differ between treatments. These data imply that EW calves, which are maintained onsite before shipping, may be more tolerant to the stressors associated with transportation and feed yard entry. Early weaned calves, managed within the system described in this study, may have improved G:F. PMID- 15753351 TI - The effect of milk intake on forage intake and growth of nursing calves. AB - Thirty-nine Holstein steer calves were assigned to one of five treatments at birth and individually fed for 200 d with milk replacer reconstituted to equal the fat and protein concentration of beef cow milk. Treatment levels were the quantities of reconstituted milk fed per day based on lactation curves, which were based on peak milk levels (PML) of 2.72, 5.44, 8.16, 10.88, and 13.6 kg/d, respectively. In addition to reconstituted milk, chopped alfalfa hay was offered ad libitum to allow for maximal voluntary forage consumption. All calves were fed a high-energy diet postweaning until they reached a similar degree of fatness in the 12th rib (4 to 5% chemical fat) as determined by ultrasound. There were differences (P < 0.05) among groups in weaning weight, preweaning ADG, age, and weight at slaughter. During the preweaning phase, there was a linear relationship (P < 0.01) for daily milk and forage DE intake; however, DE intake per unit of BW did not differ across treatments (P = 0.06). Increasing PML resulted in a linear (P < 0.01) decrease in alfalfa hay intake in the preweaning phase, and G:F increased quadratically (P < 0.01). During the postweaning phase, preweaning milk intake had no meaningful effect on postweaning ADG, but overall ADG had a linear relationship (P < 0.01) with preweaning milk level. There was no effect of PML on the 12th-rib lipid percent, marbling score, or quality grade, but protein and fat concentration in the carcass and empty BW increased linearly (P < 0.01) with PML. The group fed at 2.72 kg/d PML was 58 kg lighter (P = 0.03) and required 34 d more (P < 0.01) to reach the predetermined degree of fatness at slaughter than the group fed at 13.6 kg/d PML, suggesting that increased milk production by the dam can decrease the number of days to the slaughter weight at which a similar rib lipid concentration is reached. PMID- 15753352 TI - Breeding for societally important traits in pigs. AB - Pig breeding programs traditionally focus on the genetic improvement of production and reproduction traits that have a clear economic value. Because citizens and consumers increasingly attach value to traits that have little or no direct relationship to production costs or to the price of a product, pig breeding organizations want to pay more attention to societally important traits, such as the welfare and health of pigs, the ecological effects of pork production, and the healthiness and sensory quality of pork. Most societally important traits have an economic and a noneconomic value and are sufficiently heritable for effective genetic selection, although many genetic and phenotypic (co)-variances still have to be estimated. However, it often is not clear to a pig breeding organization how it may deal with the noneconomic value of breeding goal traits. In this study, a retrospective selection-index method is proposed to obtain the proper weights for societally important traits in the breeding goal. First, the genetic-progress space for each breeding-goal trait is explored by increasing the weights, in a stepwise manner, to each societally important trait in the breeding goal, starting from zero. Subsequently, a pig breeding organization can adopt the resulting genetic-progress scenario that it considers most sustainable or most acceptable. The weights underlying the adopted scenario are considered to be the proper breeding-goal weights. The noneconomic value of each societally important breeding-goal trait is found by deducting its economic value from its obtained weight and is thereby expressed in monetary units. In addition to obtaining weights for societally important breeding-goal traits, the proposed method offers the possibility to estimate the societal costs of selecting for economic traits only, as well as the societal benefits and the economic costs of selecting for traits with a noneconomic value. The method is therefore a useful tool for the development of sustainable breeding goals. An example has been worked out for a sow-line breeding program. PMID- 15753353 TI - Gene expression profiles in pancreatic intraepithelial neoplasia reflect the effects of Hedgehog signaling on pancreatic ductal epithelial cells. AB - Invasive pancreatic cancer is thought to develop through a series of noninvasive duct lesions known as pancreatic intraepithelial neoplasia (PanIN). We used cDNA microarrays interrogating 15,000 transcripts to identify 49 genes that were differentially expressed in microdissected early PanIN lesions (PanIN-1B/2) compared with microdissected normal duct epithelium. In this analysis, a cluster of extrapancreatic foregut markers, including pepsinogen C, MUC6, KLF4, and TFF1, was found to be up-regulated in PanIN. Up-regulation of these genes was further validated using combinations of real-time reverse transcription-PCR, in situ hybridization, and immunohistochemistry in a total of 150 early PanIN lesions from 81 patients. Identification of these gastrointestinal transcripts in human PanIN prompted assessment of other foregut markers by both semiquantitative and real-time reverse transcription-PCR, revealing similar up-regulation of Sox-2, Gastrin, HoxA5, GATA4/5/6, Villin and Forkhead 6 (Foxl1). In contrast to frequent expression of multiple gastric epithelial markers, the intestinal markers intestinal fatty acid binding protein, CDX1 and CDX2 were rarely expressed either in PanIN lesions or in invasive pancreatic cancer. Hedgehog pathway activation induced by transfection of immortalized human pancreatic ductal epithelial cells with Gli1 resulted in up-regulation of the majority of foregut markers seen in early PanIN lesions. These data show frequent up-regulation of foregut markers in early PanIN lesions and suggest that PanIN development may involve Hedgehog mediated conversion to a gastric epithelial differentiation program. PMID- 15753354 TI - Evidence for nonautonomous effect of p53 tumor suppressor in carcinogenesis. AB - Prostate, breast, and probably other epithelial tumors harbor inactivating mutations in the p53 tumor suppressor gene in the stromal cells, implying the nonautonomous action of p53 in carcinogenesis. We have tested this hypothesis by evaluating the tumorigenicity of MCF7 human breast cancer cells in severe combined immunodeficient mice that differ in their p53 status. Our results showed that, indeed, p53 ablation in the hosts reduced the latency for the development of MCF7 tumors. Furthermore, we show that heterozygous hosts frequently undergo loss of heterozygosity at the p53 locus in the tumor stroma tissue by mechanism that resembles the inactivation of p53 in primary tumors. To evaluate the impact of p53 ablation in the stromal fibroblasts, in tumorigenesis, tumors were reconstituted in mice bearing wild-type p53 alleles, by mixing MCF7 cells with fibroblasts isolated from mutant or wild-type p53 mice. Our results suggest that tumors containing p53-deficient fibroblasts developed faster and were more aggressive than their counterparts with wild-type fibroblasts, although their neoplastic component, namely MCF7 mammary carcinoma cells, was identical in both cases. These data strongly support the notion for the operation of a nonautonomous mechanism for p53 action in primary tumors and provide a mechanistic association between p53 mutations in the stromal component of epithelial tumors and carcinogenesis. PMID- 15753355 TI - Circumventing multidrug resistance in cancer by beta-galactoside binding protein, an antiproliferative cytokine. AB - We report here that beta-galactoside binding protein (betaGBP), an antiproliferative cytokine which can program cancer cells to undergo apoptosis, exhibits equal therapeutic efficacy against cancer cells that display diverse mechanisms of drug resistance and against their parental cells. The mechanisms of drug resistance in the cancer cells that we have examined include overexpression of P-glycoprotein, increased efficiency of DNA repair, and altered expression and mutation in the topoisomerase I and II enzymes. We also report that betaGBP exerted its effect by arresting the cells in S phase prior to the activation of programmed cell death. The uniquely similar profile of response to betaGBP by these drug-resistant cells and their parental cells extends the therapeutic potential of this cytokine in the treatment of cancers and offers a promising alternative to patients whose tumors are refractory to the currently available cadre of chemotherapeutic agents. PMID- 15753356 TI - Cannabinoid receptor as a novel target for the treatment of prostate cancer. AB - Cannabinoids, the active components of Cannabis sativa Linnaeus (marijuana) and their derivatives have received renewed interest in recent years due to their diverse pharmacologic activities such as cell growth inhibition, anti inflammatory effects and tumor regression. Here we show that expression levels of both cannabinoid receptors, CB1 and CB2, are significantly higher in CA-human papillomavirus-10 (virally transformed cells derived from adenocarcinoma of human prostate tissue), and other human prostate cells LNCaP, DUI45, PC3, and CWR22Rnu1 than in human prostate epithelial and PZ-HPV-7 (virally transformed cells derived from normal human prostate tissue) cells. WIN-55,212-2 (mixed CB1/CB2 agonist) treatment with androgen-responsive LNCaP cells resulted in a dose- (1-10 micromol/L) and time-dependent (24-48 hours) inhibition of cell growth, blocking of CB1 and CB2 receptors by their antagonists SR141716 (CB1) and SR144528 (CB2) significantly prevented this effect. Extending this observation, we found that WIN-55,212-2 treatment with LNCaP resulted in a dose- (1-10 micromol/L) and time dependent (24-72 hours) induction of apoptosis (a), decrease in protein and mRNA expression of androgen receptor (b), decrease in intracellular protein and mRNA expression of prostate-specific antigen (c), decrease in secreted prostate specific antigen levels (d), and decrease in protein expression of proliferation cell nuclear antigen and vascular endothelial growth factor (e). Our results suggest that WIN-55,212-2 or other non-habit-forming cannabinoid receptor agonists could be developed as novel therapeutic agents for the treatment of prostate cancer. PMID- 15753357 TI - Somatic mutations of the HER2 kinase domain in lung adenocarcinomas. AB - Mutations in the epidermal growth factor receptor gene (EGFR) in lung cancers predict for sensitivity to EGFR kinase inhibitors. HER2 (also known as NEU, EGFR2, or ERBB2) is a member of the EGFR family of receptor tyrosine kinases and plays important roles in the pathogenesis of certain human cancers, and mutations have recently been reported in lung cancers. We sequenced the tyrosine kinase domain of HER2 in 671 primary non-small cell lung cancers (NSCLC), 80 NSCLC cell lines, and 55 SCLCs and other neuroendocrine lung tumors as well as 85 other epithelial cancers (breast, bladder, prostate, and colorectal cancers) and compared the mutational status with clinicopathologic features and the presence of EGFR or KRAS mutations. HER2 mutations were present in 1.6% (11 of 671) of NSCLC and were absent in other types of cancers. Only one adenocarcinoma cell line (NCI-H1781) had a mutation. All HER2 mutations were in-frame insertions in exon 20 and target the identical corresponding region as did EGFR insertions. HER2 mutations were significantly more frequent in never smokers (3.2%, 8 of 248; P=0.02) and adenocarcinoma histology (2.8%, 11 of 394; P=0.003). In 394 adenocarcinoma cases, HER2 mutations preferentially targeted Oriental ethnicity (3.9%) compared with other ethnicities (0.7%), female gender (3.6%) compared with male gender (1.9%) and never smokers (4.1%) compared with smokers (1.4%). Mutations in EGFR, HER2, and KRAS genes were never present together in individual tumors and cell lines. The remarkable similarities of mutations in EGFR and HER2 genes involving tumor type and subtype, mutation type, gene location, and specific patient subpopulations targeted are unprecedented and suggest similar etiologic factors. EGFR, HER2, and KRAS mutations are mutually exclusive, suggesting different pathways to lung cancer in smokers and never smokers. PMID- 15753358 TI - PUMA overexpression induces reactive oxygen species generation and proteasome mediated stathmin degradation in colorectal cancer cells. AB - Increased amounts of reactive oxygen species (ROS) induce apoptosis in mammalian cells. PUMA (P53 up-regulated modulator of apoptosis), a mitochondrial proapoptotic BH3-only protein, induces rapid apoptosis through a Bax- and mitochondria-dependent pathway. However, the molecular basis of PUMA-induced apoptosis is largely not understood. Using a combination of biophysical and biochemical methods and PUMA-inducible colorectal cells, DLD-1.PUMA, we showed that (a) PUMA-induced apoptosis is dose and time dependent; (b) PUMA-induced apoptosis is directly associated with ROS generation; (c) diphenyleneiodonium chloride, a ROS blocker, or BAX-inhibiting peptide, a suppressor of BAX translocation, decreased ROS generation and apoptosis in DLD-1.PUMA cells; (d) overexpression of PUMA induced up-regulation (>1.34-fold) of peroxiredoxin 1 and down-regulation (by 25%) of stathmin through proteasome-mediated degradation; and (e) hydrogen peroxide down-regulated stathmin and disrupted the cellular microtubule network. Our findings indicate that PUMA induces apoptosis, in part, through the BAX-dependent generation of superoxide and hydrogen peroxide. ROS overproduction and oxidative stress induce proteome-wise alterations, such as stathmin degradation and disorganization of the cell microtubule network, in apoptotic cells. PMID- 15753359 TI - Positive contribution of pathogenic mutations in the mitochondrial genome to the promotion of cancer by prevention from apoptosis. AB - The role of mitochondrial dysfunction in cancer has been a subject of great interest and much ongoing investigation. Although most cancer cells harbor somatic mutations in mitochondrial DNA (mtDNA), the question of whether such mutations contribute to the promotion of carcinomas remains unsolved. Here we used trans-mitochondrial hybrids (cybrids) containing a common HeLa nucleus and mtDNA of interest to compare the role of mtDNA against the common nuclear background. We constructed cybrids with or without a homoplasmic pathogenic point mutation at nucleotide position 8,993 or 9,176 in the mtDNA ATP synthase subunit 6 gene (MTATP6) derived from patients with mitochondrial encephalomyopathy. When the cybrids were transplanted into nude mice, the MTATP6 mutations conferred an advantage in the early stage of tumor growth. The mutant cybrids also increased faster than wild type in culture. To complement the mtDNA mutations, we transfected a wild-type nuclear version of MTATP, whose codons were converted to the universal genetic codes containing a mitochondrial target sequence, into the nucleus of cybrids carrying mutant MTATP6. The restoration of MTATP slowed down the growth of tumor in transplantation. Conversely, expression of a mutant nuclear version of MTATP6 in the wild-type cybrids declined respiration and accelerated the tumor growth. These findings showed that the advantage in tumor growth depended upon the MTATP6 function but was not due to secondary nuclear mutations caused by the mutant mitochondria. Because apoptosis occurred less frequently in the mutant versus wild-type cybrids in cultures and tumors, the pathogenic mtDNA mutations seem to promote tumors by preventing apoptosis. PMID- 15753360 TI - Loss of heterozygosity patterns provide fingerprints for genetic heterogeneity in multistep cancer progression of tobacco smoke-induced non-small cell lung cancer. AB - Dilution end point loss of heterozygosity (LOH) analysis, a novel approach for the analysis of LOH, was used to evaluate allelic losses with the use of 21 highly polymorphic microsatellite markers at nine chromosomal sites most frequently affected in smoking-related non-small cell lung cancers. Allelotyping was done for bronchial epithelial cells and matching blood samples from 23 former and current smokers and six nonsmokers as well as in 33 adenocarcinomas and 25 squamous cell carcinomas (SCC) and corresponding matching blood from smokers. Major conclusions from these studies are as follows: (a) LOH at chromosomal sites 8p, 9p, 11q, and 13q (P >0.05, Fisher's exact test) are targeted at the early stages, whereas LOH at 1p, 5q, 17p, and 18q (P <0.05, Fisher's exact test) occur at the later stages of non-small cell lung cancer progression; (b) LOH at 1p, 3p, 5q, 8p, 9p, 11q, 13q, 17p, and 18q occurs in over 45% of the tobacco smokers with SCC and adenocarcinoma; (c) compared with bronchial epithelial cells from smokers, there is a significantly higher degree of LOH at 1p, 5q, and 18q in adenocarcinoma and at 1p, 3p, and 17p in SCC (P <0.05, Fisher's exact test). We propose that lung cancer progression induced by tobacco smoke occurs in a series of target gene inactivations/activations in defined modules of a global network. The gatekeeper module consists of multiple alternate target genes, which is inclusive of but not limited to genes localized to chromosomal loci 8p, 9p, 11q, and 13q. PMID- 15753361 TI - Deficiency in the catalytic subunit of DNA-dependent protein kinase causes down regulation of ATM. AB - Previous reports have suggested a connection between reduced levels of the catalytic subunit of DNA-dependent protein kinases (DNA-PKcs), a component of the nonhomologous DNA double-strand breaks end-joining system, and a reduction in ATM. We studied this possible connection in other DNA-PKcs-deficient cell types, and following knockdown of DNA-PKcs with small interfering RNA, Chinese hamster ovary V3 cells, lacking DNA-PKcs, had reduced levels of ATM and hSMG-1, but both were restored after transfection with PRKDC. Atm levels were also reduced in murine scid cells. Reduction of ATM in a human glioma cell line lacking DNA-PKcs was accompanied by defective signaling through downstream substrates, post irradiation. A large reduction of DNA-PKcs was achieved in normal human fibroblasts after transfection with two DNA-PKcs small interfering RNA sequences. This was accompanied by a reduction in ATM. These data were confirmed using immunocytochemical detection of the proteins. Within hours after transfection, a decline in PRKDC mRNA was seen, followed by a more gradual decline in DNA-PKcs protein beginning 1 day after transfection. No change in ATM mRNA was observed for 2 days post-transfection. Only after the DNA-PKcs reduction occurred was a reduction in ATM mRNA observed, beginning 2 days post-transfection. The amount of ATM began to decline, starting about 3 days post-treatment, then it declined to levels comparable to DNA-PKcs. Both proteins returned to normal levels at later times. These data illustrate a potentially important cross-regulation between the nonhomologous end-joining system for rejoining of DNA double-strand breaks and the ATM-dependent damage response network of pathways, both of which operate to maintain the integrity of the genome. PMID- 15753362 TI - Integrated array-comparative genomic hybridization and expression array profiles identify clinically relevant molecular subtypes of glioblastoma. AB - Glioblastoma, the most aggressive primary brain tumor in humans, exhibits a large degree of molecular heterogeneity. Understanding the molecular pathology of a tumor and its linkage to behavior is an important foundation for developing and evaluating approaches to clinical management. Here we integrate array-comparative genomic hybridization and array-based gene expression profiles to identify relationships between DNA copy number aberrations, gene expression alterations, and survival in 34 patients with glioblastoma. Unsupervised clustering on either profile resulted in similar groups of patients, and groups defined by either method were associated with survival. The high concordance between these separate molecular classifications suggested a strong association between alterations on the DNA and RNA levels. We therefore investigated relationships between DNA copy number and gene expression changes. Loss of chromosome 10, a predominant genetic change, was associated not only with changes in the expression of genes located on chromosome 10 but also with genome-wide differences in gene expression. We found that CHI3L1/YKL-40 was significantly associated with both chromosome 10 copy number loss and poorer survival. Immortalized human astrocytes stably transfected with CHI3L1/YKL-40 exhibited changes in gene expression similar to patterns observed in human tumors and conferred radioresistance and increased invasion in vitro. Taken together, the results indicate that integrating DNA and mRNA-based tumor profiles offers the potential for a clinically relevant classification more robust than either method alone and provides a basis for identifying genes important in glioma pathogenesis. PMID- 15753363 TI - Overexpression of soluble TRAIL induces apoptosis in human lung adenocarcinoma and inhibits growth of tumor xenografts in nude mice. AB - Recombinant adeno-associated virus 2/5 (rAAV2/5), a hybrid rAAV-2 with AAV-5 capsid, seems to be a very efficient delivery vector for the transduction of the lung adenocarcinoma cell line A549. Infection of the A549 cell line with a rAAV2/5 vector encoding the extracellular domain of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL, amino acids 114-281) resulted in secretion of soluble TRAIL (sTRAIL) and induction of apoptosis in these cells. rAAV2/5-sTRAIL mediated delivery and stable expression of sTRAIL resulted in the presence of the trimeric form of sTRAIL in sera of nude mice that were implanted with s.c. or orthotopic A549 tumors. The rAAV2/5-sTRAIL transduction of the tumors resulted in a statistically significant reduction in tumor growth and prolonged survival of the tumor-bearing animals. Primary cell culture, histologic examination of the tumors, and serum analyses showed the absence of detectable TRAIL-induced toxicity in normal tissues including the liver. The successful inhibition of lung cancer growth and the absence of detectable toxicity suggest a putative role for rAAV2/5-sTRAIL(114-281) in the therapy of lung cancer. PMID- 15753364 TI - Large-scale transcriptome analyses reveal new genetic marker candidates of head, neck, and thyroid cancer. AB - A detailed genome mapping analysis of 213,636 expressed sequence tags (EST) derived from nontumor and tumor tissues of the oral cavity, larynx, pharynx, and thyroid was done. Transcripts matching known human genes were identified; potential new splice variants were flagged and subjected to manual curation, pointing to 788 putatively new alternative splicing isoforms, the majority (75%) being insertion events. A subset of 34 new splicing isoforms (5% of 788 events) was selected and 23 (68%) were confirmed by reverse transcription-PCR and DNA sequencing. Putative new genes were revealed, including six transcripts mapped to well-studied chromosomes such as 22, as well as transcripts that mapped to 253 intergenic regions. In addition, 2,251 noncoding intronic RNAs, eventually involved in transcriptional regulation, were found. A set of 250 candidate markers for loss of heterozygosis or gene amplification was selected by identifying transcripts that mapped to genomic regions previously known to be frequently amplified or deleted in head, neck, and thyroid tumors. Three of these markers were evaluated by quantitative reverse transcription-PCR in an independent set of individual samples. Along with detailed clinical data about tumor origin, the information reported here is now publicly available on a dedicated Web site as a resource for further biological investigation. This first in silico reconstruction of the head, neck, and thyroid transcriptomes points to a wealth of new candidate markers that can be used for future studies on the molecular basis of these tumors. Similar analysis is warranted for a number of other tumors for which large EST data sets are available. PMID- 15753365 TI - Elevated serum levels of stromal-derived factor-1alpha are associated with increased osteoclast activity and osteolytic bone disease in multiple myeloma patients. AB - Multiple myeloma (MM) is an incurable plasma cell (PC) malignancy able to mediate massive destruction of the axial and craniofacial skeleton. The aim of this study was to investigate the role of the potent chemokine, stromal-derived factor 1alpha (SDF-1alpha) in the recruitment of osteoclast precursors to the bone marrow. Our studies show that MM PC produce significant levels of SDF-1alpha protein and exhibit elevated plasma levels of SDF-1alpha when compared with normal, age-matched subjects. The level of SDF-1alpha positively correlated with the presence of multiple radiological bone lesions in individuals with MM, suggesting a potential role for SDF-1alpha in osteoclast precursor recruitment and activation. To examine this further, peripheral blood-derived CD14+ osteoclast precursors were cultured in an in vitro osteoclast-potentiating culture system in the presence of recombinant human SDF-1alpha. Although failing to stimulate an increase in TRAP+, multinucleated osteoclast formation, our studies show that SDF-1alpha mediated a dramatic increase in both the number and the size of the resorption lacunae formed. The increased osteoclast motility and activation in response to SDF-1alpha was associated with an increase in the expression of a number of osteoclast activation-related genes, including RANKL, RANK, TRAP, MMP-9, CA-II, and Cathepsin K. Importantly, the small-molecule CXCR4 specific inhibitor, 4F-Benzoyl-TE14011 (T140), effectively blocked osteoclast formation stimulated by the myeloma cell line, RPMI-8226. Based on these findings, we believe that the synthesis of high levels of SDF-1alpha by MM PC may serve to recruit osteoclast precursors to local sites within the bone marrow and enhance their motility and bone-resorbing activity. Therefore, we propose that inhibition of the CXCR4-SDF-1alpha axis may provide an effective means of treatment for MM-induced osteolysis. PMID- 15753366 TI - Osteoprotegerin in prostate cancer bone metastasis. AB - Osteoprotegerin (OPG), a critical regulator of osteoclastogenesis, is expressed by prostate cancer cells, and OPG levels are increased in patients with prostate cancer bone metastases. The objective of this study was to investigate the effects of OPG overexpression on prostate cancer cells and prostate cancer/bone cell interactions in vitro and in vivo. OPG-transfected C4-2 cells expressed 8.0 ng OPG per mL per 10(6) cells, whereas no OPG was detected in the media of C4-2 cells transfected with a control plasmid. OPG overexpressed by C4-2 cells protected these cells from tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis and decreased osteoclast formation. Subcutaneous OPG-C4 2 and pcDNA-C4-2 tumors exhibited similar growth and take-rate characteristics. However, when grown in bone, tumor volume was decreased in OPG-C4-2 versus pcDNA C4-2 (P=0.0017). OPG expressed by C4-2 cells caused increases in bone mineral density (P=0.0074) and percentage of trabecular bone volume (P=0.007), and decreases in numbers of osteoblasts and osteoclasts when compared with intratibial pcDNA-C4-2 tumors (P=0.003 and P=0.019, respectively). In summary, our data show that increased expression of OPG in C4-2 cells does not directly affect proliferation of prostate cancer cells but indirectly decreases growth of C4-2 tumors in the bone environment. Our data also show that OPG expressed by C4 2 cells inhibits bone lysis associated with C4-2 bone metastasis, which results in net increases in bone volume. We therefore hypothesize that OPG expressed in prostate cancer patient bone metastases may be at least partially responsible for the osteoblastic character of most prostate cancer bone lesions. PMID- 15753367 TI - Differential effects of neuropeptide Y on the growth and vascularization of neural crest-derived tumors. AB - Neuropeptide Y (NPY) is a sympathetic neurotransmitter recently found to be potently angiogenic and growth promoting for endothelial, vascular smooth muscle and neuronal cells. NPY and its cognate receptors, Y1, Y2 and Y5, are expressed in neural crest-derived tumors; however, their role in regulation of growth is unknown. The effect of NPY on the growth and vascularization of neuroendocrine tumors was tested using three types of cells: neuroblastoma, pheochromocytoma, and Ewing's sarcoma family of tumors (ESFT). The tumors varied in expression of NPY receptors, which was linked to differential functions of the peptide. NPY stimulated proliferation of neuroblastoma cells via Y2/Y5Rs and inhibited ESFT cell growth by Y1/Y5-mediated apoptosis. In both tumor types, NPY receptor antagonists altered basal growth levels, indicating a regulatory role of autocrine NPY. In addition, the peptide released from the tumor cells stimulated endothelial cell proliferation, which suggests its paracrine angiogenic effects. In nude mice xenografts, exogenous NPY stimulated growth of neuroblastoma tumors, whereas it increased apoptosis and reduced growth of ESFT. However, in both tumors, NPY treatment led to an increase in tumor vascularization. Taken together, this is the first report of NPY being a growth-regulatory factor for neuroendocrine tumors, acting both by autocrine activation of tumor cell proliferation or apoptosis and by angiogenesis. NPY and its receptors may become targets for novel approaches in the treatment of these diseases, directed against both tumor cell proliferation and angiogenesis. PMID- 15753368 TI - RET-familial medullary thyroid carcinoma mutants Y791F and S891A activate a Src/JAK/STAT3 pathway, independent of glial cell line-derived neurotrophic factor. AB - The RET proto-oncogene encodes a receptor tyrosine kinase whose dysfunction plays a crucial role in the development of several neural crest disorders. Distinct activating RET mutations cause multiple endocrine neoplasia type 2A (MEN2A), type 2B (MEN2B), and familial medullary thyroid carcinoma (FMTC). Despite clear correlations between the mutations found in these cancer syndromes and their phenotypes, the molecular mechanisms connecting the mutated receptor to the different disease phenotypes are far from completely understood. Luciferase reporter assays in combination with immunoprecipitations, and Western and immunohistochemistry analyses were done in order to characterize the signaling properties of two FMTC-associated RET mutations, Y791F and S891A, respectively, both affecting the tyrosine kinase domain of the receptor. We show that these RET FMTC mutants are monomeric receptors which are autophosphorylated and activated independently of glial cell line-derived neurotrophic factor. Moreover, we show that the dysfunctional signaling properties of these mutants, when compared with wild-type RET, involve constitutive activation of signal transducers and activators of transcription 3 (STAT3). Furthermore, we show that STAT3 activation is mediated by a signaling pathway involving Src, JAK1, and JAK2, differing from STAT3 activation promoted by RET(C634R) which was previously found to be independent of Src and JAKs. Three-dimensional modeling of the RET catalytic domain suggested that the structural changes promoted by the respective amino acids substitutions lead to a more accessible substrate and ATP-binding monomeric conformation. Finally, immunohistochemical analysis of FMTC tumor samples support the in vitro data, because nuclear localized, Y705-phosphorylated STAT3, as well as a high degree of RET expression at the plasma membrane was observed. PMID- 15753369 TI - Kaposi's sarcoma-associated herpesvirus viral IFN regulatory factor 1 inhibits transforming growth factor-beta signaling. AB - Kaposi's sarcoma-associated herpesvirus, also called human herpesvirus 8, has been implicated in the pathogenesis of Kaposi's sarcoma, body cavity-based primary effusion lymphoma, and some forms of multicentric Castleman's disease. The Kaposi's sarcoma-associated herpesvirus open reading frame K9 encodes viral IFN regulatory factor 1 (vIRF1), which functions as a repressor of IFN-mediated signal transduction. vIRF1 expression in NIH 3T3 cells leads to transformation and consequently induces malignant fibrosarcoma in nude mice, suggesting that vIRF1 is a strong oncoprotein. Here, we show that vIRF1 inhibited transforming growth factor-beta (TGF-beta) signaling via its targeting of Smad proteins. vIRF1 suppressed TGF-beta-mediated transcription and growth arrest. vIRF1 directly interacted with both Smad3 and Smad4, resulting in inhibition of their transactivation activity. Studies using vIRF1 deletion mutants showed that the central region of vIRF1 was required for vIRF1 association with Smad3 and Smad4 and that this region was also important for inhibition of TGF-beta signaling. In addition, we found that vIRF1 interfered with Smad3-Smad4 complex formation and inhibited Smad3/Smad4 complexes from binding to DNA. These results indicate that vIRF1 inhibits TGF-beta signaling via interaction with Smads. In addition, the data indicate the TGF-beta pathway is an important target for viral oncoproteins. PMID- 15753370 TI - An expression signature classifies chemotherapy-resistant pediatric osteosarcoma. AB - Osteosarcoma is the most common malignant bone tumor in children. Osteosarcoma patients who respond poorly to chemotherapy are at a higher risk of relapse and adverse outcome. Therefore, it was the aim of this study to identify prognostic factors at the time of diagnosis to characterize the genes predictive of poor survival outcome and to identify potential novel therapeutic targets. Expression profiling of 30 osteosarcoma diagnostic biopsy samples, 15 with inferior necrosis following induction chemotherapy (Huvos I/II) and 15 with superior necrosis following induction chemotherapy (Huvos III/IV), was conducted using Affymetrix U95Av2 oligonucleotide microarrays. One hundred and four genes were found to be statistically significant and highly differentially expressed between Huvos I/II and III/IV patients. Statistically significant genes were validated on a small independent cohort comprised of osteosarcoma xenograft tumor samples. Markers of Huvos I/II response predominantly were gene products involved in extracellular matrix (ECM) microenvironment remodeling and osteoclast differentiation. A striking finding was the significant decrease in osteoprotegerin, an osteoclastogenesis inhibitory factor. Additional genes involved in osteoclastogenesis and bone resorption, which were statistically different, include annexin 2, SMAD, PLA2G2A, and TGFbeta1. ECM remodeling genes include desmoplakin, SPARCL1, biglycan, and PECAM. Gene expression of select genes involved in tumor progression, ECM remodeling, and osteoclastogenesis were validated via quantitative reverse transcription-PCR in an independent cohort. We propose that osteosarcoma tumor-driven changes in the bone microenvironment contribute to the chemotherapy-resistant phenotype and offer testable hypotheses to potentially enhance therapeutic response. PMID- 15753371 TI - Skin carcinoma arising from donor cells in a kidney transplant recipient. AB - The incidence of skin cancer is increased in transplant recipients. UV radiation, papillomaviruses, and immunosuppression participate in the pathogenesis of these tumors. In addition, donor cells may leave the grafted organ, reach peripheral tissues and either induce immune phenomena or possibly take part in tissue remodeling. Herein, we investigated the possible involvement of donor cells in the development of skin tumors in kidney allograft recipients. We analyzed a series of 48 malignant and benign cutaneous tumors developing in 14 females who had been grafted with a male kidney. The number of male cells was measured on microdissected material by quantitative PCR for Y chromosome. In the samples with high levels of male cells, fluorescent in situ hybridization (FISH) with X and Y probes and/or immuno-FISH with anticytokeratin antibodies were carried out. Male cells were detected in 5/15 squamous cell carcinomas and Bowen disease (range 4 180 copies), 3/5 basal cell carcinomas (91-645), 6/11 actinic keratosis (7-102), 2/4 keratoacanthoma (22-41), and 2/5 benign cutaneous lesions (14-55). In a basal cell carcinoma specimen with a high number of male cells, FISH showed that most cells within the tumoral buds were XY. In this lesion, immuno-FISH showed the presence of XY cytokeratin-positive cells indicating that the tumor nests contained male keratinocytes. In contrast, in other female transplants, male cells present in the tumors were not epithelial. In conclusion, stem cells originating from a grafted kidney may migrate to the skin, differentiate, or fuse as keratinocytes that could, rarely, undergo cancer transformation. PMID- 15753372 TI - Adoptive transfer of tumor-reactive transforming growth factor-beta-insensitive CD8+ T cells: eradication of autologous mouse prostate cancer. AB - Transforming growth factor (TGF)-beta is a potent immunosuppressant. Overproduction of TGF-beta by tumor cells may lead to tumor evasion from the host immune surveillance and tumor progression. The present study was conducted to develop a treatment strategy through adoptive transfer of tumor-reactive TGF-beta insensitive CD8+ T cells. The mouse TRAMP-C2 prostate cancer cells produced large amounts of TGF-beta1 and were used as an experimental model. C57BL/6 mice were primed with irradiated TRAMP-C2 cells. CD8+ T cells were isolated from the spleen of primed animals, were expanded ex vivo, and were rendered TGF-beta insensitive by infecting with a retrovirus containing dominant-negative TGF-beta type II receptor. Results of in vitro cytotoxic assay revealed that these CD8+ T cells showed a specific and robust tumor-killing activity against TRAMP-C2 cells but were ineffective against an irrelevant tumor line, B16-F10. To determine the in vivo antitumor activity, recipient mice were challenged with a single injection of TRAMP-C2 cells for a period up to 21 days before adoptive transfer of CD8+ T cells was done. Pulmonary metastasis was either eliminated or significantly reduced in the group receiving adoptive transfer of tumor-reactive TGF-beta insensitive CD8+ T cells. Results of immunofluorescent studies showed that only tumor-reactive TGF-beta-insensitive CD8+ T cells were able to infiltrate into the tumor and mediate apoptosis in tumor cells. Furthermore, transferred tumor reactive TGF-beta-insensitive CD8+ T cells were able to persist in tumor-bearing hosts but declined in tumor-free animals. These results suggest that adoptive transfer of tumor-reactive TGF-beta-insensitive CD8+ T cells may warrant consideration for cancer therapy. PMID- 15753373 TI - Multidrug transporter ABCG2 prevents tumor cell death induced by the epidermal growth factor receptor inhibitor Iressa (ZD1839, Gefitinib). AB - Iressa (ZD1839, Gefitinib), used in clinics to treat non-small cell lung cancer patients, is a tyrosine kinase receptor inhibitor that leads to specific decoupling of epidermal growth factor receptor (EGFR) signaling. Recent data indicate that Iressa is especially effective in tumors with certain EGFR mutations; however, a subset of these tumors does not respond to Iressa. In addition, certain populations have an elevated risk of side effects during Iressa treatment. The human ABCG2 (BCRP/MXR/ABCP) transporter causes cancer drug resistance by actively extruding a variety of cytotoxic drugs, and it functions physiologically to protect our tissues from xenobiotics. Importantly, ABCG2 modifies absorption, distribution, and toxicity of several pharmacologic agents. Previously, we showed that ABCG2 displays a high-affinity interaction with several tyrosine kinase receptor inhibitors, including Iressa. Here, we show that the expression of ABCG2, but not its nonfunctional mutant, protects the EGFR signaling-dependent A431 tumor cells from death on exposure to Iressa. This protection is reversed by the ABCG2-specific inhibitor, Ko143. These data, reinforced with cell biology and biochemical experiments, strongly suggest that ABCG2 can actively pump Iressa. Therefore, variable expression and polymorphisms of ABCG2 may significantly modify the antitumor effect as well as the absorption and tissue distribution of Iressa. PMID- 15753374 TI - Identification of metastasis-associated receptor tyrosine kinases in non-small cell lung cancer. AB - Development of distant metastasis after tumor resection is the leading cause of death in early-stage non-small cell lung cancer (NSCLC). Receptor tyrosine kinases (RTK) are involved in tumorigenesis but only few RTKs have been systematically studied in NSCLC. Here, we provide quantitative real-time reverse transcription-PCR expression data of all RTKs (n=56) in primary tumors of 70 patients with early-stage (I-IIIA) NSCLC. Overall, 33 RTKs were expressed in at least 25% of the patients. Several RTKs were significantly expressed higher in tumors that ultimately metastasized. The hazard risk for metastasis development in stage I/II disease was increased at least 3-fold for tumors with high expression levels of insulin receptor, neurotrophic tyrosine receptor kinase 1, epidermal growth factor receptor, ERBB2, ERBB3, platelet-derived growth factor receptor beta, fibroblast growth factor receptor 1, or leukocyte tyrosine kinase. Relative risks were reduced 3-fold by expression of EPHB6 or DKFZ1. Three members of the epidermal growth factor receptor family were associated with a high risk of metastasis, emphasizing the validity of our data. High ERBB3 expression was significantly associated with decreased survival. Taken together, our genome-wide RTK expression map uncovered the previously unknown value of several RTKs as potential markers for prognosis and metastasis prediction in early-stage NSCLC. The identified RTKs represent promising novel candidates for further functional analyses. PMID- 15753375 TI - E-cadherin suppression accelerates squamous cell carcinoma progression in three dimensional, human tissue constructs. AB - We studied the link between loss of E-cadherin-mediated adhesion and acquisition of malignant properties in three-dimensional, human tissue constructs that mimicked the initial stages of squamous cell cancer progression. Suppression of E cadherin expression in early-stage, skin-derived tumor cells (HaCaT-II-4) was induced by cytoplasmic sequestration of beta-catenin upon stable expression of a dominant-negative E-cadherin fusion protein (H-2Kd-Ecad). In monolayer cultures, expression of H-2Kd-Ecad resulted in decreased levels of E-cadherin, redistribution of beta-catenin to the cytoplasm, and complete loss of intercellular adhesion when compared with control II-4 cells. This was accompanied by a 7-fold decrease in beta-catenin-mediated transcription and a 12 fold increase in cell migration. In three-dimensional constructs, E-cadherin deficient tissues showed disruption of architecture, loss of adherens junctional proteins from cell contacts, and focal tumor cell invasion. Invasion was linked to activation of matrix metalloproteinase (MMP)-mediated degradation of basement membrane in H-2Kd-Ecad-expressing tissue constructs that was blocked by MMP inhibition (GM6001). Quantitative reverse transcription-PCR showed a 2.5-fold increase in MMP-2 and an 8-fold increase in MMP-9 in cells expressing the H-2Kd Ecad fusion protein when compared with controls, and gel zymography showed increased MMP protein levels. Following surface transplantation of three dimensional tissues, suppression of E-cadherin expression greatly accelerated tumorigenesis in vivo by inducing a switch to high-grade carcinomas that resulted in a 5-fold increase in tumor size after 4 weeks. Suppression of E-cadherin expression and loss of its function fundamentally modified squamous cell carcinoma progression by activating a highly invasive, aggressive tumor phenotype, whereas maintenance of E-cadherin prevented invasion in vitro and limited tumor progression in vivo. PMID- 15753376 TI - p38 regulates cyclooxygenase-2 in human mammary epithelial cells and is activated in premalignant tissue. AB - The immediate-early gene, cyclooxygenase-2 (COX-2), is induced in a variety of inflammatory and neoplastic processes and is believed to play an important role in tumorigenesis. In this study, we identify an important upstream regulatory pathway of COX-2 expression in variant human mammary epithelial cells (vHMEC), which has been shown to exhibit phenotypes important for malignancy. We find that the stress-activated kinase, p38, is phosphorylated and activated in vHMEC compared with HMEC and is responsible for the expression of COX-2 in vHMEC as cells grow in culture. Furthermore in this capacity, p38 acts to stabilize the COX-2 transcript rather than activate COX-2 transcription. Inhibition of p38 kinase, using a chemical inhibitor, down-regulates COX-2 and decreases cell survival. Examination of archived tissue from women with ductal carcinoma in situ reveals epithelial cells that not only overexpress COX-2 but also have an abundance of activated phospho-p38 in the nucleus and cytoplasm, mirroring the expression observed in vitro. These epithelial cells are found within premalignant lesions as well as in fields of morphologically normal tissue that surround the lesions. In contrast, low phospho-p38 staining was observed in the majority of normal tissue obtained from reduction mammoplasty. These data help define the regulation of COX-2 expression in early carcinogenesis and provide alternative candidates for targeted prevention of COX-2-induced phenotypes and breast cancer. PMID- 15753377 TI - Epigenetic up-regulation of C-C chemokine receptor 7 and C-X-C chemokine receptor 4 expression in melanoma cells. AB - Histone deacetylation and DNA methylation establish epigenetic modifications, which through chromatin remodeling may result in gene silencing. We hypothesized that chemokine receptors C-C chemokine receptor 7 (CCR7) and C-X-C chemokine receptor 4 (CXCR4) on melanoma cells undergo epigenetic regulation. We investigated whether a histone deacetylase inhibitor and a demethylating agent influence CCR7 and CXCR4 expression on melanoma cells. Initially, microarray analysis was done to screen changes in chemokine receptor expression on melanoma cells after treatment with trichostatin A (TSA) and 5-Aza-2-deoxycytidine (5 Aza). CCR7 and CXCR4 mRNA expression were uniformly altered and selected for further investigation. Quantitative real-time reverse transcription-PCR assay, immunohistochemistry, and Western blot analysis were used to assess changes in mRNA and protein expression induced by TSA and 5-Aza in melanoma lines. Cell migration assays were conducted to assess the effects of altered CCR7 and CXCR4 expression on cell function. Treatment with TSA or 5-Aza increased gene expression of both CCR7 and CXCR4 in melanoma lines. TSA was the strongest enhancer. With combined treatment, CCR7 and CXCR4 mRNA expression was also up regulated. Immunohistochemistry after combined treatment showed enhanced staining of both CCR7 and CXCR4 compared with control cells. Melanoma cell migration in TSA- and 5-Aza-treated cells was 7- and 2-fold higher than control cells for CCR7 and CXCR4, respectively. In summary, a histone deacetylase inhibitor and a demethylating agent up-regulated CCR7 and CXCR4 expression on melanoma cells. This increase in chemokine receptor expression correlated with functional activity. Most importantly, we have identified an epigenetic mechanism that may endogenously regulate chemokine receptor expression on melanoma cells. PMID- 15753378 TI - Transitional cell hyperplasia and carcinomas in urinary bladders of transgenic mice with keratin 5 promoter-driven cyclooxygenase-2 overexpression. AB - The inducible form of cyclooxygenase (COX), COX-2, is up-regulated in many epithelial cancers and its prostaglandin products increase proliferation, enhance angiogenesis, and inhibit apoptosis in several tissues. Pharmacologic inhibition and genetic deletion studies showed a marked reduction of tumor development in colon and skin. COX-2 has also been strongly implicated in urinary bladder cancer primarily by studies with nonselective COX- and COX-2-selective inhibitors. We now show that forced expression of COX-2, under the control of a keratin 5 promoter, is sufficient to cause transitional cell hyperplasia (TCH) in 17% and 75% of the heterozygous and homozygous transgenic lines, respectively, in an age dependent manner. TCH was strongly associated with inflammation, primarily nodules of B lymphocytes; some T cells and macrophage infiltration were also observed. Additionally, transitional cell carcinoma was observed in approximately 10% of the K5.COX-2 transgenic mice; no TCH or transitional cell carcinoma was observed in wild-type bladders. Immunohistochemistry for vascular proliferation and vascular endothelial growth factor showed significant increases above that in wild-type urinary bladders. Our results suggest that overexpression of COX-2 is sufficient to cause hyperplasia and carcinomas in the urinary bladder. Therefore, inhibition of COX-2 should continue to be pursued as a potential chemopreventive and therapeutic strategy. PMID- 15753379 TI - Iterative microarray and RNA interference-based interrogation of the SRC-induced invasive phenotype. AB - Src kinase has long been recognized as a factor in the progression of colorectal cancer and seems to play a specific role in the development of the metastatic phenotype. In spite of numerous studies conducted to elucidate the exact role of Src in cancer progression, downstream targets of Src remain poorly understood. Gene expression profiling has permitted the identification of large sets of genes that may be functionally interrelated but it is often unclear as to which molecular pathways they belong. Here we have developed an iterative approach to experimentally reconstruct a network of gene activity regulated by Src and contributing to the invasive phenotype. Our strategy uses a combination of phenotypic anchoring of gene expression profiles and loss-of-function screening by way of RNA-mediated interference. Using a panel of human colon cancer cell lines exhibiting differential Src-specific activity and invasivity, we identify the first two levels of gene transcription responsible for the invasive phenotype, where first-tier genes are controlled by Src activity and the second tier genes are under the influence of the first tier. Specifically, perturbation of first-tier gene activity by either pharmacologic inhibition of Src or RNA mediated interference-directed knockdown leads to a loss of invasivity and decline of second-tier gene activity. The targeting of first-tier genes may be bypassed altogether because knockdown of second-tier genes led to a similar loss of invasive potential. In this manner, numerous members of a "transcriptional cascade" pathway for metastatic activity have been identified and functionally validated. PMID- 15753380 TI - Prostaglandin E2 enhances intestinal adenoma growth via activation of the Ras mitogen-activated protein kinase cascade. AB - A large body of clinical, genetic, and biochemical evidence indicates that cyclooxygenase-2 (COX-2), a key enzyme for prostanoid biosynthesis, contributes to the promotion of colorectal cancer. COX-2-derived prostaglandin E2 (PGE2) is the most abundant prostaglandin found in several gastrointestinal malignancies. Although PGE2 enhances intestinal adenoma growth in Apcmin mice, the mechanism(s) by which it accelerates tumor growth is not completely understood. Here we investigated how PGE2 promotes intestinal tumor growth and the signaling pathways responsible for its effects. We observed that PGE2 treatment leads to increased epithelial cell proliferation and induces COX-2 expression in intestinal adenomas. Furthermore, we show that PGE2 regulation of COX-2 expression is mediated by activation of a Ras-mitogen-activated protein kinase signaling cascade. One intriguing finding is that COX-2-derived PGE2 mimics the effects of constitutively active Ras through a self-amplifying loop that allows for a distinct growth advantage. PMID- 15753381 TI - Specificity of the methylation-suppressed A isoform of candidate tumor suppressor RASSF1 for microtubule hyperstabilization is determined by cell death inducer C19ORF5. AB - Isoform-specific epigenetic silencing of RASSF1A (3p21.3) by promoter-specific CpG island hypermethylation occurs at high frequency in human tumors, whereas the closely related product of the same gene, RASSF1C, continues to be expressed. Both isoforms in isolation exhibit tumor suppressor properties and we show here similar cellular locations on mitochondria and microtubules, paclitaxel-like microtubule hyperstabilization, disruption of mitosis, and interaction with C19ORF5. We show both have identical but distinct sequence domains for microtubule association and hyperstabilization. C19ORF5 is a hyperstabilized microtubule-specific binding protein of which accumulation causes mitochondrial aggregation and cell death. We report herein that when A or C isoforms of RASSF1 are coexpressed with C19ORF5, the unique N-terminal sequence of RASSF1C prevents it from hyperstabilizing microtubules. This confers specificity on RASSF1A in microtubule hyperstabilization and accumulation of C19ORF5 on microtubules and could underlie a specific effect of hypermethylation-suppressed RASSF1A in tumor suppression. PMID- 15753382 TI - The growth arrest function of the human oncoprotein mouse double minute-2 is disabled by downstream mutation in cancer cells. AB - We have reported earlier that ectopic expression of mouse double minute-2 (MDM2) induces G1 arrest in normal cells. To explain occasional overexpression of MDM2 in cancer cells, we searched for deletion or substitution mutation in the growth suppressor domains of MDM2 in several breast cancer cell lines that overexpress the oncoprotein. Our results suggest the absence of alteration (deletion or substitution) in the open reading frame of MDM2 transcripts in such cells. Because the breast cancer cell line MCF-7 overexpresses MDM2, we isolated the full-length MDM2 transcript from this cell line. The MDM2 cDNA synthesized from transcripts isolated from MCF-7 cells induced inhibition of G1 to S phase transition in normal human diploid cells such as WI38, suggesting that the genetic alterations in breast cancer cells that overexpress MDM2 disable the growth arrest function of the oncoprotein. Consistently, overexpression of full length MDM2 in MCF-7 cells over its high endogenous level did not inhibit G1-S transition efficiently. Although MDM2 overexpression was accompanied by CDK4 overexpression or absence of cdk4 inhibitor p16 in most breast cancer cells, we found remarkably high levels of cyclin A rather than cyclin E in these cells. Ectopic expression of cyclin A released MDM2-mediated inhibition of G1-S transition in normal human diploid WI38 cells. We propose that cancer cells expressing high levels of cyclin A escape MDM2-mediated G1 arrest, which may account for a selective growth advantage over normal cells. PMID- 15753383 TI - Androgens up-regulate the insulin-like growth factor-I receptor in prostate cancer cells. AB - In this study, we show that androgens up-regulate insulin-like growth factor-I receptor (IGF-IR) expression and sensitize prostate cancer cells to the biological effects of IGF-I. Both dihydrotestosterone and the synthetic androgen R1881 induced an approximately 6-fold increase in IGF-IR expression in androgen receptor (AR)-positive prostate cancer cells LNCaP. In accordance with IGF-IR up regulation, treatment with the nonmetabolizable androgen R1881 sensitized LNCaP cells to the mitogenic and motogenic effects of IGF-I, whereas an IGF-IR blocking antibody effectively inhibited these effects. By contrast, these androgens did not affect IGF-IR expression in AR-negative prostate cancer cells PC-3. Reintroduction of AR into PC-3 cells by stable transfection restored the androgen effect on IGF-IR up-regulation. R1881-induced IGF-IR up-regulation was partially inhibited by the AR antagonist Casodex (bicalutamide). Two other AR antagonists, cyproterone acetate and OH-flutamide, were much less effective. Androgen-induced IGF-IR up-regulation was not dependent on AR genomic activity, because two AR mutants, AR-C619Y and AR-C574R, devoid of DNA binding activity and transcriptional activity were still able to elicit IGF-IR up-regulation in HEK293 kidney cells in response to androgens. Moreover, androgen-induced IGF-IR up regulation involves the activation of the Src-extracellular signal-regulated kinase pathway, because it was inhibited by both the Src inhibitor PP2 and the MEK-1 inhibitor PD98059. The present observations strongly suggest that AR activation may stimulate prostate cancer progression through the altered IGF-IR expression and IGF action. Anti-androgen therapy may be only partially effective, or almost ineffective, in blocking important biological effects of androgens, such as activation of the IGF system. PMID- 15753384 TI - ErbB2 promotes Src synthesis and stability: novel mechanisms of Src activation that confer breast cancer metastasis. AB - Activation of Src kinase plays important roles in the development of many neoplasias. Most of the previous Src studies focused on the deregulation of Src kinase activity. The deregulated Src protein synthesis and stability in mediating malignant phenotypes of cancer cells, however, have been neglected. While investigating the signal transduction pathways contributing to ErbB2-mediated metastasis, we found that ErbB2-activated breast cancer cells that had higher metastatic potentials also had increased Src activity compared with ErbB2 low expressing cells. The increased Src activity in ErbB2-activated cells paralleled higher Src protein levels, whereas Src RNA levels were not significantly altered. Our studies revealed two novel mechanisms that are involved in Src protein up regulation and activation by ErbB2: (a) ErbB2 increased Src translation through activation of the Akt/mammalian target of rapamycin/4E-BP1 pathway and (b) ErbB2 increased Src stability most likely through the inhibition of the calpain protease. Furthermore, inhibition of Src activity by a Src-specific inhibitor, PP2, or a Src dominant-negative mutant dramatically reduced ErbB2-mediated cancer cell invasion in vitro and metastasis in an experimental metastasis animal model. Together, activation of ErbB2 and downstream signaling pathways can lead to increased Src protein synthesis and decreased Src protein degradation resulting in Src up-regulation and activation, which play critical roles in ErbB2-mediated breast cancer invasion and metastasis. PMID- 15753385 TI - Induction of claudin-4 by nonsteroidal anti-inflammatory drugs and its contribution to their chemopreventive effect. AB - Nonsteroidal anti-inflammatory drugs (NSAID) have shown chemopreventive effects in both preclinical and clinical studies; however, the precise molecular mechanism governing this response remains unclear. We used DNA microarray techniques to search for genes whose expression is induced by the NSAID indomethacin in human gastric carcinoma (AGS) cells. Among identified genes, we focused on those related to tight junction function (claudin-4, claudin-1, and occludin), particularly claudin-4. Induction of claudin-4 by indomethacin was confirmed at both mRNA and protein levels. NSAIDs, other than indomethacin (diclofenac and celecoxib), also induced claudin-4. All of the tested NSAIDs increased the intracellular Ca2+ concentration. Other drugs that increased the intracellular Ca2+ concentration (thapsigargin and ionomycin) also induced claudin-4. Furthermore, an intracellular Ca2+ chelator [1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid] inhibited the indomethacin dependent induction of claudin-4. These results strongly suggest that induction of claudin-4 by indomethacin is mediated through an increase in the intracellular Ca2+ concentration. Overexpression of claudin-4 in AGS cells did not affect cell growth or the induction of apoptosis by indomethacin. On the other hand, addition of indomethacin or overexpression of claudin-4 inhibited cell migration. Colony formation in soft agar was also inhibited. Suppression of claudin-4 expression by small interfering RNA restored the migration activity of AGS cells in the presence of indomethacin. Based on these results, we consider that the induction of claudin-4 and other tight junction-related genes by NSAIDs may be involved in the chemopreventive effect of NSAIDs through the suppression of anchorage independent growth and cell migration. PMID- 15753387 TI - Functional proteomic screen identifies a modulating role for CD44 in death receptor-mediated apoptosis. AB - Apoptotic evasion is a hallmark of cancer and its resistance to chemotherapeutic drugs. Identification of cellular proteins that mediate apoptotic programs is a critical step toward the development of therapeutics aimed at overcoming apoptosis resistance. We developed an innovative high-throughput screen to identify proteins that modulate Fas ligand-mediated apoptosis using fluorophore assisted light inactivation (HTS-FALIpop). The FALI protein knockdown strategy was coupled to a caspase activity assay with the ability to detect both proapoptotic and antiapoptotic surface molecules expressed by HT-1080 human fibrosarcoma cells. FALI of the Fas receptor (Fas/CD95) using a fluorescein conjugated anti-Fas antibody abrogated Fas ligand-mediated caspase activation. Ninety-six single-chain variable fragment antibodies (scFv), selected for binding to the surface of HT-1080 cells, were screened by HTS-FALIpop. Three of the scFvs caused decreases in caspase induction after FALI of their protein targets. One of the targets of these positive scFvs was identified as CD44 and was validated by performing FALI using a CD44-specific monoclonal antibody, which resulted in similar protection from Fas apoptosis. CD44-targeted FALI was antiapoptotic in multiple human cancer cell lines, including both Fas signaling type I and II cells, and was also protective against other ligands of the tumor necrosis factor death receptor family. FALI of CD44 inhibited formation and activation of the death-inducing signaling complex, suggesting that CD44 regulates Fas at the cell surface. This mechanism of death receptor regulation represents a novel means of apoptosis modulation that could be exploited by pharmacologic agents. PMID- 15753386 TI - Activin A suppresses neuroblastoma xenograft tumor growth via antimitotic and antiangiogenic mechanisms. AB - The tumor suppressor function of activin A, together with our findings that activin A is an inhibitor of angiogenesis, which is down-regulated by the N-MYC oncogene, prompted us to investigate in more detail its role in the malignant transformation process of neuroblastomas. Indeed, neuroblastoma cells with restored activin A expression exhibited a diminished proliferation rate and formed smaller xenograft tumors with reduced vascularity, whereas lung metastasis rate remained unchanged. In agreement with the decreased vascularity of the xenograft tumors, activin A inhibited several crucial angiogenic responses of cultured endothelial cells, such as proteolytic activity, migration, and proliferation. Endothelial cell proliferation, activin A, or its constitutively active activin receptor-like kinase 4 receptor (ALK4T206D), increased the expression of CDKN1A (p21), CDKN2B (p15), and CDKN1B (p27) CDK inhibitors and down-regulated the expression of vascular endothelial growth factor receptor-2, the receptor of a key angiogenic factor in cancer. The constitutively active forms of SMAD2 and SMAD3 were both capable of inhibiting endothelial cell proliferation, whereas the dominant-negative forms of SMAD3 and SMAD4 released the inhibitory effect of activin A on endothelial cell proliferation by only 20%. Thus, the effects of activin A on endothelial cell proliferation seem to be conveyed via the ALK4/SMAD2-SMAD3 pathways, however, non-SMAD cascades may also contribute. These results provide novel information regarding the role of activin A in the malignant transformation process of neuroblastomas and the molecular mechanisms involved in regulating angiogenesis thereof. PMID- 15753388 TI - Imatinib mesylate inhibits Leydig cell tumor growth: evidence for in vitro and in vivo activity. AB - Leydig cell tumors are usually benign tumors of the male gonad. However, if the tumor is malignant, no effective treatments are currently available. Leydig cell tumors express platelet-derived growth factor (PDGF), kit ligand and their respective receptors, PDGFR and c-kit. We therefore evaluated the effects of imatinib mesylate (imatinib), a selective inhibitor of the c-kit and PDGFR tyrosine kinases, on the growth of rodent Leydig tumor cell lines in vivo and in vitro, and examined, in human Leydig cell tumor samples, the expression of activated PDGFR and c-kit and the mutations in exons of the c-kit gene commonly associated with solid tumors. Imatinib caused concentration-dependent decreases in the viability of Leydig tumor cell lines, which coincided with apoptosis and inhibition of proliferation and ligand-stimulated phosphorylation of c-kit and PDGFRs. Mice bearing s.c. allografts of a Leydig tumor cell line treated with imatinib p.o., had an almost complete inhibition of tumor growth, less tumor cell proliferation, increased apoptosis, and a lesser amount of tumor-associated mean vessel density compared with controls. No drug-resistant tumors appeared during imatinib treatment but tumors regrew after drug withdrawal. Human Leydig cell tumors showed an intense expression of the phosphorylated form of c-kit and a less intense expression of phosphorylated PDGFRs. No activating mutations in common regions of mutation of the c-kit gene were found. Our studies suggest that Leydig cell tumors might be a potential target for imatinib therapy. PMID- 15753389 TI - Targeting beta-transducin repeat-containing protein E3 ubiquitin ligase augments the effects of antitumor drugs on breast cancer cells. AB - beta-Transducin repeat-containing proteins (beta-TrCP) serve as substrate recognition component of E3 ubiquitin ligases that control stability of important regulators of cell cycle and signal transduction. beta-TrCP function is essential for the induction of nuclear factor kappaB transcriptional activities, which play a key role in proliferation and survival of cancer cells and are often constitutively up-regulated in human breast cancers. Here we show that inhibition of beta-TrCP either by RNAi approach or by forced expression of a dominant negative beta-TrCP mutant suppresses growth and survival of human breast cancer cells. In addition, inhibition of beta-TrCP augments the antiproliferative effects of anticancer drugs such as doxorubicin, tamoxifen, and paclitaxel on human mammary tumor cells. These data provide the proof of principle that targeting beta-TrCP might be beneficial for anticancer therapies. PMID- 15753390 TI - Real-time detection of gene expression in cancer cells using molecular beacon imaging: new strategies for cancer research. AB - Development of novel approaches for quantitative analysis of gene expression in intact tumor cells should provide new means for cancer detection and for studying the response of cancer cells to biological and therapeutic reagents. We developed procedures for detecting the levels of expression of multiple genes in fixed as well as viable cells using molecular beacon imaging technology. We found that simultaneous delivery of molecular beacons targeting survivin and cyclin D1 mRNAs produced strong fluorescence in breast cancer but not in normal breast cells. Importantly, fluorescence intensity correlated well with the level of gene expression in the cells detected by real-time reverse transcription-PCR or Western blot analysis. We further show that molecular beacons can detect changes of survivin gene expression in viable cancer cells following epidermal growth factor stimulation, docetaxel treatment, and overexpression of p53 gene. Thus, molecular beacon imaging is a simple and specific method for detecting gene expression in cancer cells. It has great potential for cancer detection and drug development. PMID- 15753391 TI - Activation of p53 by MDM2 antagonists can protect proliferating cells from mitotic inhibitors. AB - Recent studies have shown that activation of cell cycle checkpoints can protect normal proliferating cells from mitotic inhibitors by preventing their entry into mitosis. These studies have used genotoxic agents that act, at least in part, by activation of the p53 pathway. However, genotoxic drugs are known also to have p53-independent activities and could affect the sensitivity of tumor cells to antimitotic agents. Recently, we have developed the first potent and selective small-molecule inhibitors of the p53-MDM2 interaction, the nutlins, which activate the p53 pathway only in cells with wild-type but not mutant p53. Using these compounds, we show that p53 activation leads to G1 and G2 phase arrest and can protect cells from mitotic block and apoptosis caused by paclitaxel. Pretreatment of HCT116 and RKO colon cancer cells (wild-type p53) or primary human fibroblasts (1043SK) with nutlins for 24 hours followed by incubation with paclitaxel for additional 48 hours did not increase significantly their mitotic index and protected the cells from the cytotoxicity of paclitaxel. Cancer cells with mutant p53 (MDA-MB-435) responded to the same treatment with mitotic arrest and massive apoptosis. These results have two major implications for cancer therapy. First, p53-activating therapies may have antagonistic effect when combined with mitotic poisons. Second, pretreatment with MDM2 antagonists before chemotherapy of tumors with mutant p53 may offer a partial protection to proliferating normal tissues. PMID- 15753392 TI - Monoclonal anti-idiotype antibody 6G6.C4 fused to GM-CSF is capable of breaking tolerance to carcinoembryonic antigen (CEA) in CEA-transgenic mice. AB - Internal image anti-idiotypic antibodies are capable of mimicking tumor associated antigens and thus may serve as surrogate for vaccination strategies in cancer patients. The monoclonal antibody (mAb) 6G6.C4 mimics an epitope specific for the human carcinoembryonic antigen (CEA) and generates a CEA-specific response (Ab3) in various experimental animals. In humans, however, 6G6.C4 only yields a very limited humoral anti-CEA reaction presumably due to tolerance against the CEA autoantigen. In this study, we investigated the CEA-specific Ab3 response in mice transgenic for the human CEA and tested whether the antigen tolerance could be overcome by fusing a recombinant single-chain variable fragment of 6G6.C4 (scFv6G6.C4) to the murine granulocyte macrophage colony stimulating factor (GM-CSF). Like mAb 6G6.C4, the fusion protein (scFv6G6.C4/GM CSF) retained binding to the CEA-specific idiotype mAb T84.66. Also, scFv6G6.C4/GM-CSF was biologically active as measured by proliferation of the GM CSF-dependent murine FDC-P1 cells in vitro. After immunization with the scFv6G6.C4/GM-CSF fusion protein, CEA-transgenic animals showed significantly enhanced Ab3 antibody responses to scFv6G6.C4 (P=0.005) and to CEA (P=0.012) compared with the scFV6G6.C4 alone. Sera from mice immunized with the fusion protein specifically recognized CEA in Western blot analyses with no cross reaction to CEA-related antigens. Finally, the Ab3 antisera detected single CEA expressing tumor cells in suspension as shown by flow cytometry. Taken together, these data show in a model antigenically related to the human system that vaccination with scFv6G6.C4/GM-CSF improves vaccination against an endogenous tumor-associated antigen resulting in a highly specific humoral Ab3 response in vivo that is capable of bind single circulating CEA-positive tumor cells. PMID- 15753393 TI - The excitatory amino acid transporter-2 induces apoptosis and decreases glioma growth in vitro and in vivo. AB - Accumulating evidence suggests that glutamate plays a key role in the proliferation and invasion of glioblastoma tumors. Astrocytic tumors have been shown to release glutamate at high levels, which may stimulate tumor cell proliferation and motility via activation of glutamate receptors. Excess glutamate has also been found to facilitate tumor invasion by causing excitotoxic damage to normal brain thereby paving a pathway for tumor migration. Results from tissue microarray analyses showed decreased excitatory amino acid transporter-2 (EAAT-2) expression in high-grade glial tumors compared with low-grade astrocytomas and normal brain. EAAT-2 expression was inversely correlated with tumor grade, implicating its potential role in glial tumor progression, which was reflected by an undetectable level of EAAT-2 protein in glioma cell lines. In this study, we sought to investigate the effect of reconstituted EAAT-2 on glioma cell growth in vitro and in vivo by adenoviral-mediated gene transfer. Infection of glioma cells with Ad-EAAT-2 resulted in a physiologic level of functional EAAT 2, and a subsequent dose-dependent reduction in cell proliferation in all glioma cell lines tested compared with controls. Interestingly, results from analyses of Annexin V staining, detection of poly(ADP-ribose)polymerase cleavage and caspase 3 activation all indicated that Ad-EAAT-2 infection elicited apoptosis in glioma cells. Ex vivo experiments in nude mice showed a total suppression of tumor growth at sites that received Ad-EAAT-2-infected cells. Collectively, our results uncovered a new function of EAAT-2 in controlling glioma proliferation. Further studies will improve our knowledge of the role of glutamate in glioma growth and may provide useful prognostic information and alternative therapeutic targets for the treatment of glioma. PMID- 15753394 TI - Gene therapy for prostate cancer by controlling adenovirus E1a and E4 gene expression with PSES enhancer. AB - PSES is a chimeric enhancer containing enhancer elements from prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA) genes that are prevalently expressed in androgen-independent prostate cancers. PSES shows strong activity equivalent to cytomegalovirus (CMV) promoter, specifically in PSA/PSMA positive prostate cancer cells, the major cell types in prostate cancer in the absence of androgen. We developed a recombinant adenovirus (AdE4PSESE1a) by placing adenoviral E1a and E4 genes under the control of the bidirectional enhancer PSES and enhanced green fluorescent protein gene for the purpose of intratumoral virus tracking under the control of CMV promoter. Because of PSES being very weak in nonprostatic cells, including HEK293 and HER911 that are frequently used to produce recombinant adenovirus, AdE4PSESE1a can only be produced in the HER911E4 cell line which expresses both E1 and E4 genes. AdE4PSESE1a showed similar viral replication and tumor cell killing activities to wild-type adenovirus in PSA/PSMA-positive prostate cancer cells. The viral replication and tumor cell killing activities were dramatically attenuated in PSA/PSMA-negative cells. To test whether AdE4PSESE1a could be used to target prostate tumors in vivo, CWR22rv s.c. tumors were induced in nude mice and treated with AdE4PSESE1a via intratumoral and tail vein injection. Compared to tumors treated with control virus, the growth of CWR22rv tumors was dramatically inhibited by AdE4PSESE1a via tail vein injection or intratumoral injection. These data show that adenoviral replication can be tightly controlled in a novel fashion by controlling adenoviral E1a and E4 genes simultaneously with a single enhancer. PMID- 15753395 TI - Cyclooxygenase-2 inhibits novel ginseng metabolite-mediated apoptosis. AB - Recently, a novel intestinal bacterial metabolite of ginseng protopanaxadiol saponins, i.e., 20-O-(beta-D-glucopyranosyl)-20(S)-protopanaxadiol (IH-901), has been reported to induce apoptosis in a variety of cancer cells. Here we show a differential effect of IH-901 on several cell types. Exposure to IH-901 for 48 hours at a supposedly subapoptotic concentration of 40 mumol/L led to both apoptotic cell death and G1 arrest in Hep3B cells, but only resulted in G1 arrest in MDA-MB-231, Hs578T, and MKN28 cells. Additionally, the treatment of MDA-MB 231, but not of Hep3B, with IH-901 up-regulated cyclooxygenase-2 (COX-2) mRNA (2 hours) and protein (6 hours), and enhanced the production of prostaglandin E2. In MDA-MB-231 cells, IH-901 induced the sustained activation of extracellular signal regulated kinase (ERK), whereas inhibition of mitogen-activated protein/ERK kinase blocked IH-901-mediated COX-2 induction and resulted in apoptosis, suggesting the involvement of an ERK-COX-2 pathway. Combined treatment with IH 901 and nonsteroidal anti-inflammatory drugs inhibited COX-2 enzyme and induced apoptosis in MDA-MB-231 and Hs578T cells. Adenovirus-mediated COX-2 small interfering RNAs also effectively inhibited COX-2 protein expression and enhanced IH-901-mediated apoptosis without inhibiting ERK 1/2 phosphorylation, thus providing direct evidence that COX-2 is an antiapoptotic molecule. Moreover, IH 901-mediated G1 arrest resulted from an increase in p27Kip1 mRNA and protein expression followed by a decrease in CDK2 kinase activity that was concurrent with the hypophosphorylation of Rb and p130. In conclusion, IH-901 induced both G1 arrest and apoptosis, and this apoptosis could be inhibited by COX-2 induction. PMID- 15753396 TI - Role of glycogen synthase kinase 3beta in rapamycin-mediated cell cycle regulation and chemosensitivity. AB - The mammalian target of rapamycin is a serine-threonine kinase that regulates cell cycle progression. Rapamycin and its analogues inhibit the mammalian target of rapamycin and are being actively investigated in clinical trials as novel targeted anticancer agents. Although cyclin D1 is down-regulated by rapamycin, the role of this down-regulation in rapamycin-mediated growth inhibition and the mechanism of cyclin D1 down-regulation are not well understood. Here, we show that overexpression of cyclin D1 partially overcomes rapamycin-induced cell cycle arrest and inhibition of anchorage-dependent growth in breast cancer cells. Rapamycin not only decreases endogenous cyclin D1 levels but also decreases the expression of transfected cyclin D1, suggesting that this is at least in part caused by accelerated proteolysis. Indeed, rapamycin decreases the half-life of cyclin D1 protein, and the rapamycin-induced decrease in cyclin D1 levels is partially abrogated by proteasome inhibitor N-acetyl-leucyl-leucyl-norleucinal. Rapamycin treatment leads to an increase in the kinase activity of glycogen synthase kinase 3beta (GSK3beta), a known regulator of cyclin D1 proteolysis. Rapamycin-induced down-regulation of cyclin D1 is inhibited by the GSK3beta inhibitors lithium chloride, SB216763, and SB415286. Rapamycin-induced G1 arrest is abrogated by nonspecific GSK3beta inhibitor lithium chloride but not by selective inhibitor SB216763, suggesting that GSK3beta is not essential for rapamycin-mediated G1 arrest. However, rapamycin inhibits cell growth significantly more in GSK3beta wild-type cells than in GSK3beta-null cells, suggesting that GSK3beta enhances rapamycin-mediated growth inhibition. In addition, rapamycin enhances paclitaxel-induced apoptosis through the mitochondrial death pathway; this is inhibited by selective GSK3beta inhibitors SB216763 and SB415286. Furthermore, rapamycin significantly enhances paclitaxel induced cytotoxicity in GSK3beta wild-type but not in GSK3beta-null cells, suggesting a critical role for GSK3beta in rapamycin-mediated paclitaxel sensitization. Taken together, these results show that GSK3beta plays an important role in rapamycin-mediated cell cycle regulation and chemosensitivity and thus significantly potentiates the antitumor effects of rapamycin. PMID- 15753397 TI - Regulation of p38 phosphorylation and topoisomerase IIalpha expression in the B cell lymphoma line Jiyoye by CD26/dipeptidyl peptidase IV is associated with enhanced in vitro and in vivo sensitivity to doxorubicin. AB - CD26 is a Mr 110,000 surface-bound glycoprotein with diverse functional properties, including having a key role in normal T-cell physiology and the development of certain cancers. In this article, we show that surface expression of CD26, especially its intrinsic dipeptidyl peptidase IV (DPPIV) enzyme activity, results in enhanced topoisomerase IIalpha level in the B-cell line Jiyoye and subsequent in vitro sensitivity to doxorubicin-induced apoptosis. In addition, we show that expression of CD26/DPPIV is associated with increased phosphorylation of p38 and its upstream regulators mitogen-activated protein kinase kinase 3/6 and apoptosis signal-regulating kinase 1 and that p38 signaling pathway plays a role in the regulation of topoisomerase IIalpha expression. Besides demonstrating that CD26 effect on topoisomerase IIalpha and doxorubicin sensitivity is applicable to cell lines of both B-cell and T-cell lineages, the potential clinical implication of our work lies with the fact that we now show for the first time that our in vitro results can be extended to a severe combined immunodeficient mouse model. Our findings that CD26 expression can be an in vivo marker of tumor sensitivity to doxorubicin treatment may lead to future treatment strategies targeting CD26/DPPIV for selected human cancers in the clinical setting. Our article thus characterizes the biochemical linkage among CD26, p38, and topoisomerase IIalpha while providing evidence that CD26-associated topoisomerase IIalpha expression results in greater in vitro and in vivo tumor sensitivity to the antineoplastic agent doxorubicin. PMID- 15753398 TI - Jasmonates: novel anticancer agents acting directly and selectively on human cancer cell mitochondria. AB - We reported previously that jasmonates can kill human cancer cells. Many chemotherapeutic drugs induce mitochondrial membrane permeability transition, membrane depolarization, osmotic swelling, and release of cytochrome c, involving the opening of the permeability transition pore complex (PTPC). Because jasmonates exert their cytotoxic effects independent of transcription, translation, and p53 expression, we hypothesized that these compounds may act directly on mitochondria. Mitochondrial membrane depolarization was determined by flow cytometry, and cytochrome c release by Western blotting. Mitochondria were isolated by mechanical lysis and differential centrifugation. Cytotoxicity was measured by a tetrazolium-based assay, and mitochondrial swelling by spectrophotometry. Jasmonates induced membrane depolarization and cytochrome c release in intact human cancer cell lines. Jasmonates induced swelling in mitochondria isolated from Hep 3B hepatoma cells, but not in mitochondria isolated from 3T3 nontransformed cells or from normal lymphocytes, in a PTPC mediated manner. Methyl jasmonate induced the release of cytochrome c from mitochondria isolated from cancer cell lines in a PTPC-mediated manner, but not from mitochondria isolated from normal lymphocytes. A correlation was found between cytotoxicity of methyl jasmonate and the percentage of leukemic cells in the blood of patients with chronic lymphocytic leukemia (CLL). Jasmonates induced membrane depolarization in CLL cells, and swelling and release of cytochrome c in mitochondria isolated from these cells. In conclusion, jasmonates act directly on mitochondria derived from cancer cells in a PTPC-mediated manner, and could therefore bypass premitochondrial apoptotic blocks. Jasmonates are promising candidates for the treatment of CLL and other types of cancer. PMID- 15753399 TI - Inactivation of the mitogen-activated protein kinase pathway as a potential target-based therapy in ovarian serous tumors with KRAS or BRAF mutations. AB - Activation of mitogen-activated protein kinase (MAPK) occurs in response to various growth stimulating signals and as a result of activating mutations of the upstream regulators, KRAS and BRAF, which can be found in many types of human cancer. To investigate the roles of MAPK activation in tumors harboring KRAS or BRAF mutations, we inactivated MAPK in ovarian tumor cells using CI-1040, a compound that selectively inhibits MAPK kinase, an upstream regulator of MAPK and thus prevents MAPK activation. Profound growth inhibition and apoptosis were observed in CI-1040-treated tumor cells with mutations in either KRAS or BRAF in comparison with the ovarian cancer cells containing wild-type sequences. Long serial analysis of gene expression identified several differentially expressed genes in CI-1040-treated MPSC1 cells harboring an activating mutation in BRAF (V599L). The most striking changes were down-regulation of cyclin D1, COBRA1, and transglutaminase-2 and up-regulation of tumor necrosis factor-related apoptosis induced ligand, thrombospondin-1, optineurin, and palladin. These patterns of gene expression were validated in other CI-1040-treated tumor cells based on quantitative PCR. Constitutive expression of cyclin D1 partially reversed the growth inhibitory effect of CI-1040 in MPSC1 cells. Our findings indicate that an activated MAPK pathway is critical in tumor growth and survival of ovarian tumors with KRAS or BRAF mutations and suggest that the CI-1040 induced phenotypes depend on the mutational status of KRAS and BRAF in ovarian tumors. PMID- 15753400 TI - Bcl-2 overexpression enhances tumor-specific T-cell survival. AB - Although immunotherapy based on the adoptive transfer of tumor-specific T lymphocytes has been shown to result in dramatic clinical responses in some patients, the relatively low levels of engraftment and persistence of the adoptively transferred cells may limit these responses in many patients. In an attempt to develop strategies for prolonging the survival of adoptively transferred T cells, we have carried out studies in which T cells obtained from healthy donors as well as tumor-specific T cells were transduced with a retrovirus expressing the human Bcl-2 gene. Our results indicate that these transduced T cells overexpress Bcl-2, are resistant to death, and have a survival advantage following interleukin-2 withdrawal compared with control T cells transduced with a retrovirus expressing green fluorescent protein. Tumor-specific T cells overexpressing Bcl-2 maintained their ability to specifically recognize and respond to target cells. Furthermore, we show that adoptive immunotherapy of an established B16 tumor can be significantly enhanced by overexpressing Bcl-2 in melanoma-specific T-cell receptor transgenic T cells. Our data suggest that adoptive immunotherapy approaches to the treatment of cancer patients may be enhanced using Bcl-2-modified tumor-reactive T cells. PMID- 15753401 TI - Potent selection of antigen loss variants of B16 melanoma following inflammatory killing of melanocytes in vivo. AB - We have reported that i.d. injection of plasmids encoding hsp70 and a suicide gene transcriptionally targeted to melanocytes generates specific proinflammatory killing of melanocytes. The resulting CD8+ T cell response eradicates systemically established B16 tumors. Here, we studied the consequences of that CD8+ T cell response on the phenotype of preexisting tumor. In suboptimal protocols, the T cell response selected B16 variants, which grow extremely aggressively, are amelanotic and have lost expression of the tyrosinase and tyrosinase-related protein 2 (TRP-2) antigens. However, expression of other melanoma-associated antigens, such as gp100, was not affected. Antigen loss could be reversed by long-term growth in culture away from immune-selective pressures or within 96 hours by treatment with the demethylating agent 5-azacytidine (5 Aza). When transplanted back into syngeneic animals, variants were very poorly controlled by further vaccination. However, a combination of vaccination with 5 Aza to reactivate antigen expression in tumors in situ generated highly significant improvements in therapy over treatment with vaccine or 5-Aza alone. These data show that inflammatory killing of normal cells activates a potent T cell response targeted against a specific subset of self-antigens but can also lead to the immunoselection of tumor variants. Moreover, our data indicate that emergence of antigen loss variants may often be due to reversible epigenetic mechanisms within the tumor cells. Therefore, combination therapy using vaccination and systemic treatment with 5-Aza or other demethylating agents may have significant therapeutic benefits for antitumor immunotherapy. PMID- 15753402 TI - CD4+ T cell-mediated antigen-specific immunotherapy in a mouse model of cervical cancer. AB - A major agenda for tumor immunology is the generation of specific immune responses leading to the destruction of incipient and frank neoplasia. In this report, we show that a novel HPV16 E7 fusion protein can produce objective therapeutic responses against incipient cervical cancer in genetically engineered mice that express in the cervix the HPV16 early region genes implicated as causative agents in human cervical cancer. Although nonresponsive toward the HPV16 E7 oncoprotein in the CD8+ T-cell compartment by virtue of MHC haplotype, the mice were capable of mounting an induced CD4+ T-cell response against E7, and in addition developed spontaneous anti-E7 antibodies. HPV16/CD4-/- mice showed increased tumor burden indicative of CD4-mediated immune surveillance. Seeking to enhance the CD4 response, we immunized mice bearing incipient cervical cancer with a recombinant protein fusing E7 with a mycobacterial heat shock protein. The incidences of cervical carcinoma and of high-grade dysplasia (CIN 3) were consequently reduced by comparison to control mice. Thus, an HPV16 E7 immunogen holds promise for noninvasive treatment and prevention of human cervical cancer. PMID- 15753403 TI - Activated marrow-infiltrating lymphocytes effectively target plasma cells and their clonogenic precursors. AB - A major limitation of adoptive immunotherapy is the availability of T cells specific for both terminally differentiated tumor cells and their clonogenic precursors. We show here that marrow-infiltrating lymphocytes (MILs) recognize myeloma cells after activation with anti-CD3/CD28 beads with higher frequency than activated peripheral blood lymphocytes from the same patients. Furthermore, activated MILs target both the terminally differentiated CD138+ plasma cells and the myeloma precursor as shown by profound inhibition in a tumor clonogenic assay. The presence of antigen in the marrow microenvironment seems to be important for the maintenance of tumor specificity. Taken together, these results highlight the intrinsic tumor specificity of MILs and describe a novel approach for the generation of tumor-specific T-cell populations suitable for adoptive immunotherapy of multiple myeloma. PMID- 15753404 TI - Bax and Bak are required for apoptosis induction by sulforaphane, a cruciferous vegetable-derived cancer chemopreventive agent. AB - Sulforaphane, a constituent of many edible cruciferous vegetables, including broccoli, effectively suppresses proliferation of cancer cells in culture and in vivo by causing apoptosis induction, but the sequence of events leading to cell death is poorly defined. Here, we show that multidomain proapoptotic Bcl-2 family members Bax and Bak play a critical role in apoptosis induction by sulforaphane. This conclusion is based on the following observations: (a) sulforaphane treatment caused a dose- and time-dependent increase in the protein levels of both Bax and Bak and conformational change and mitochondrial translocation of Bax in SV40-transformed mouse embryonic fibroblasts (MEF) derived from wild-type mice to trigger cytosolic release of apoptogenic molecules (cytochrome c and Smac/DIABLO), activation of caspase-9 and caspase-3, and ultimately cell death; (b) MEFs derived from Bax or Bak knockout mice resisted cell death by sulforaphane, and (c) MEFs derived from Bax and Bak double knockout mice exhibited even greater protection against sulforaphane-induced cytochrome c release, caspase activation, and apoptosis compared with wild-type or single knockout cells. Interestingly, sulforaphane treatment also caused a dose- and time-dependent increase in the protein level of Apaf-1 in wild-type, Bax-/-, and Bak-/- MEFs but not in double knockout, suggesting that Bax and Bak might regulate sulforaphane-mediated induction of Apaf-1 protein. A marked decline in the protein level of X-linked inhibitor of apoptosis on treatment with sulforaphane was also observed. Thus, it is reasonable to postulate that sulforaphane-induced apoptosis is amplified by a decrease in X-linked inhibitor of apoptosis level, which functions to block cell death by inhibiting activities of caspases. In conclusion, the results of the present study indicate that Bax and Bak proteins play a critical role in initiation of cell death by sulforaphane. PMID- 15753405 TI - Efficacy of trastuzumab. PMID- 15753406 TI - The first seizure in childhood: don't just do something, stand there! PMID- 15753407 TI - Neonatal seizures: after all these years we still love what doesn't work. PMID- 15753408 TI - Coming of age: the use of immunomodulatory therapy in children with multiple sclerosis. PMID- 15753409 TI - Idiopathic REM sleep behavior disorder: toward a better nosologic definition. AB - REM sleep behavior disorder (RBD) is a parasomnia characterized by a lack of motor inhibition during REM sleep leading to potentially harmful dream-enacting behaviors. RBD affects mainly older men and its prevalence in the general population is estimated to be around 0.5%. RBD may be idiopathic or associated with other neurologic disorders. A strong association between RBD and alpha synucleinopathies has been recently observed, with the parasomnia often heralding the clinical onset of the neurodegenerative disease. The idiopathic form accounts for up to 60% of the cases reported in the three largest series of patients with RBD. Small clinical follow-up studies revealed that a proportion of these patients will eventually develop a parkinsonian syndrome or a dementia of Lewy bodies type in the years following the RBD diagnosis, while some patients will never show other neurologic signs within several decades from the RBD onset. Recent studies have looked at neurophysiologic and neuropsychological functions in idiopathic RBD and have found evidences of CNS dysfunction during both wakefulness and sleep. An impairment of the cortical activity, specific neuropsychological deficits, and signs of autonomic dysfunction have been observed in a variable proportion of these patients, challenging the concept of idiopathic RBD. Identifying subjects with a high risk of developing a neurodegenerative process may be crucial in order to develop early intervention strategies. PMID- 15753410 TI - Removal of epileptogenic sequences from video material: the role of color. AB - BACKGROUND: After Pokemon viewing triggered an epidemic of seizures in Japan, many efforts have been made to design safety guidelines and systems to protect subjects with photosensitivity. The authors developed a new method based upon nonlinear diffusion techniques capable of filtering the epileptogenic content of a video sequence related to color without altering its spatial and luminance content. METHODS: The authors showed to 25 photosensitive patients (18 women, mean age: 22 years) the original Pokemon sequence and a modified one in an ABBA protocol using two television (TV) sets (100 and 50 Hz). RESULTS: Twenty-three patients had a photoparoxysmal response (PPR) according to Waltz classification with at least one of the scenes. The modified sequence triggered fewer and less severe PPRs than the original version in both TVs (p < 0.001). Original sequences elicited generalized PPRs in 56.5% of the trials for the 50 Hz TV and in 41.3% for the 100 Hz TV, whereas modified sequences elicited these responses in only 8.7% (50 Hz) and 4.3% (100 Hz TV) of the trials (p < 0.001). Sensitivity to the modified version on the 50 Hz TV correlated with pattern sensitivity (p < 0.05). CONCLUSION: Specific manipulations of the color modulation-depth could be enough to decrease dramatically the risk of triggering seizures in susceptible subjects exposed to provocative visual scenes. This new method can be implemented in protective devices able to filter out the epileptogenic video sequences in which color plays a fundamental role while leaving intact the spatial content, frequency, and average luminance. PMID- 15753411 TI - Effects of topiramate and gabapentin on cognitive abilities in healthy volunteers. AB - OBJECTIVE: To evaluate the cognitive effects of topiramate (TPM) and gabapentin (GBP). METHODS: Forty healthy volunteers were randomized to a 12-week course of TPM, GBP, or placebo. Doses were gradually escalated over 10 weeks to a maximum of 400 mg/day of TPM or 3,600 mg/day of GBP or to the highest tolerated dose. Subjects were interviewed and examined biweekly. Cognitive testing was performed prior to initiating the drug and again 12 weeks later, at least 2 weeks after achieving plateau dosing. For each subject and cognitive measure, test-retest Z scores were calculated based on regression equations derived from 73 healthy volunteers. Group comparisons utilized the Wilcoxon test. RESULTS: There were significant TPM vs GBP and TPM vs placebo differences in test-retest Z scores for four of six target cognitive measures (Digit Symbol, Story Recall, Selective Reminding, Controlled Oral Word Association), always indicating worse retest performance for subjects receiving TPM. Overall, 12 of 24 cognitive measures were similarly affected. TPM effects were large, and several target measures averaged >2 SD of negative change. One measure was significantly affected by GBP. CONCLUSIONS: Topiramate (TPM) impaired cognitive test performance, whereas gabapentin had minimal effects. The effects of TPM were of sufficient magnitude potentially to affect daily and occupational function. PMID- 15753412 TI - Reading impairment in the neuronal migration disorder of periventricular nodular heterotopia. AB - OBJECTIVE: To define the behavioral profile of periventricular nodular heterotopia (PNH), a malformation of cortical development that is associated with seizures but reportedly normal intelligence, and to correlate the results with anatomic and clinical features of this disorder. METHODS: Ten consecutive subjects with PNH, all with epilepsy and at least two periventricular nodules, were studied with structural MRI and neuropsychological testing. Behavioral results were statistically analyzed for correlation with other features of PNH. RESULTS: Eight of 10 subjects had deficits in reading skills despite normal intelligence. Processing speed and executive function were also impaired in some subjects. More marked reading difficulties were seen in subjects with more widely distributed heterotopia. There was no correlation between reading skills and epilepsy severity or antiepileptic medication use. CONCLUSION: The neuronal migration disorder of periventricular nodular heterotopia is associated with an impairment in reading skills despite the presence of normal intelligence. PMID- 15753413 TI - A double-blind, randomized trial of IV immunoglobulin treatment in acute optic neuritis. AB - OBJECTIVE: To investigate if IV immunoglobulin (IVIG) treatment in the acute phase of optic neuritis (ON) could improve visual outcome and reduce MRI disease activity 6 months after onset of ON. METHODS: Sixty-eight patients with ON were randomized within 4 weeks from onset of symptoms. Thirty-four patients were randomized to IVIG 0.4 g/kg body wt, and 34 patients were randomized to placebo. Infusions were given at days 0, 1, 2, 30, and 60. Contrast sensitivity, visual acuity, and color vision were measured at baseline and after 1 week, 1 month, and 6 months. Pattern reversal visual evoked potential studies and gadolinium enhanced MRI were performed at baseline and after 1 and 6 months. Clinical relapses during follow-up were recorded. RESULTS: There was no difference in the primary outcome, contrast sensitivity after 6 months, between patients randomized to treatment with IVIG or placebo. In addition, there was no significant difference in the secondary outcome measures, improvement in the visual function measures and MRI, at any time during follow-up. At baseline, a significantly higher number of patients in the IVIG group had one or more enhancing lesions on MRI and IVIG-treated patients had a significantly higher number of enhancing lesions on MRI than patients treated with placebo. No difference was found in number of patients with one or more enhancing lesions or number of enhancing lesions in subsequent scans between treatment groups. Number of relapses was equal in the two treatment groups during follow-up. CONCLUSIONS: There was no effect of IV immunoglobulin (IVIG) on long-term visual function following acute optic neuritis, nor was there an effect of IVIG treatment in reducing latency on visual evoked potentials and thus preserving function of axons of the optic nerve. PMID- 15753414 TI - Risedronate therapy for prevention of hip fracture after stroke in elderly women. AB - BACKGROUND: There is a high incidence of hip fractures in patients with hemiplegic stroke. Bone mineral density (BMD) is decreased in the hemiplegic side in patients after stroke, correlating with the degree of paralysis and of hypovitaminosis D. OBJECTIVE: To evaluate the efficacy of risedronate in reducing the severity of osteoporosis and in decreasing the risk of hip fractures in elderly women following an acute stroke. METHODS: This was a 12-month, randomized, double blind, placebo-controlled trial. In a prospective study of stroke patients, 187 patients received a daily dose of 2.5 mg risedronate for 12 months, and the remaining 187 received placebo. Incidence of hip fracture was compared between the two groups at the endpoint of the study. RESULTS: Seven patients sustained hip fractures on the hemiplegic side in the placebo group, and one hip fracture occurred in the risedronate group (p = 0.0360; OR = 7.0). BMD increased by 1.5% and decreased by 4.9% in the risedronate group and placebo group (p < 0.0001). Urinary deoxypyridinoline, a bone resorption marker, decreased by 53.4% in the risedronate group and increased by 35.8% in the placebo group. CONCLUSION: Treatment with risedronate increases bone mineral density in elderly women following an acute stroke and prevents hip fractures. PMID- 15753415 TI - Timing of TIAs preceding stroke: time window for prevention is very short. AB - BACKGROUND: Patients with TIA are at increased risk of ischemic stroke and require preventive treatment. However, clinical guidelines differ on how urgently patients should be assessed. OBJECTIVE: To determine the potential consequences of delays in investigation and treatment, the authors studied the timing of TIAs preceding ischemic stroke. METHODS: The authors studied patients who presented with a recent ischemic stroke and had a preceding TIA in two population-based studies (Oxford Vascular Study [OXVASC]; Oxfordshire Community Stroke Project [OCSP]) and two randomized trials (UK TIA Aspirin Trial [UK-TIA]; European Carotid Surgery Trial [ECST]). RESULTS: Of 2,416 patients who had presented with an ischemic stroke, 549 (23%) gave a history of a preceding TIA (18% in OXVASC, 15% in OCSP, 23% in UK-TIA, 26% in ECST). Where a preceding TIA had occurred, the timing was highly consistent across the studies, with 17% occurring on the day of the stroke, 9% on the previous day, and 43% at some point during the 7 days prior to the stroke. No clinical characteristics or vascular risk factors identified patients in whom there was a close temporal association between TIA and stroke. CONCLUSION: In patients presenting with ischemic stroke, TIAs occur most often during the hours and days immediately preceding the stroke. PMID- 15753416 TI - The prognostic value of domain-specific cognitive abilities in acute first-ever stroke. AB - OBJECTIVE: To evaluate the prognostic value of domain-specific cognitive abilities in acute stroke with respect to long-term cognitive and functional outcome in addition to neurologic and demographic predictors. METHODS: The authors evaluated 168 patients within the first 3 weeks after first-ever stroke. The prevalence of neuropsychological impairment was calculated vs 75 matched healthy controls. The authors also recorded demographic data, vascular risk factors, lesion characteristics, and clinical factors at admission. Independent predictor variables associated with long-term cognitive impairment (assessed with a follow-up neuropsychological examination) and functional impairment (assessed with the modified Barthel Index and the Frenchay Activities Index) were identified with stepwise multiple logistic regression. Areas under receiver operator characteristic curves were used to compare the predictive value of three models, i.e., a standard medical model, a purely cognitive model, and a model consisting of both medical and cognitive predictors. RESULTS: Thirty-one percent of patients showed long-term cognitive impairment. Basic and instrumental ADL disturbances remained present in 19% and 24% of patients. Domain-specific cognitive functioning predicted cognitive and functional outcome better than any other variable. Moreover, the prediction of instrumental ADL functioning improved when cognitive predictors were added to the standard medical model (p < 0.05). Impairments in abstract reasoning and executive functioning were independent predictors of long-term cognitive impairment. Inattention and perceptual disorders were more important in predicting long-term functional impairment. CONCLUSION: Domain-specific cognitive abilities in the early phase of stroke are excellent independent predictors of long-term cognitive and functional outcome. PMID- 15753417 TI - Ischemic optic neuropathy and carotid dissection. PMID- 15753418 TI - "Diffusion-clinical mismatch" is associated with potential for early recovery of aphasia. AB - BACKGROUND: Diffusion-perfusion mismatch (perfusion-weighted imaging [PWI] abnormality minus diffusion-weighted imaging [DWI] abnormality) can identify candidates for acute stroke intervention, but PWI is often not obtainable. The authors hypothesized that language tests can predict volume of hypoperfusion, and thus mismatch, in acute left hemisphere stroke, and that the estimated mismatch can predict potential for early recovery of language. METHODS: A consecutive series of 81 patients with acute left hemisphere ischemic stroke underwent language testing within 1 day of MRI scans. Volumes of abnormality on PWI and DWI were measured without knowledge of language scores. Using tests that correlated well with PWI abnormality (oral naming and repetition), the authors computed an estimated PWI abnormality (ePWI) for each patient from a linear regression model and derived a diffusion-clinical percent mismatch ([ePWI-DWI/ePWI] x 100). The authors then tested the hypothesis that patients with > or =20% diffusion clinical mismatch have a greater chance of short-term improvement in language by examining scores of the 23 patients with repeat testing within 1 week. RESULTS: Within-group comparisons: patients with > or =20% diffusion-clinical mismatch showed improvement in total language score within 1 week (Wilcoxon signed rank: p < 0.02), whereas patients without mismatch did not. Across-group comparison: patients with > or =20% mismatch showed more short-term improvement in language scores than those without mismatch (Mann-Whitney U: p < 0.05). CONCLUSIONS: Tests of oral naming or repetition can be used in patients with acute left hemisphere stroke to estimate perfusion-weighted imaging abnormality and compute a diffusion clinical mismatch that may predict potential for short-term language improvement. PMID- 15753419 TI - Mild cognitive impairment is related to Alzheimer disease pathology and cerebral infarctions. AB - OBJECTIVES: To examine the extent to which persons with mild cognitive impairment have intermediate levels of Alzheimer disease (AD) pathology, cerebral infarcts, and Lewy body disease. METHODS: A total of 180 Catholic clergy participating in the Religious Orders Study underwent annual detailed evaluation and brain autopsy. Blocks of midfrontal, superior temporal, medial temporal lobe, inferior parietal, entorhinal cortex, hippocampus, and substantia nigra were paraffin embedded, and sectioned at 6 mum. Cortical neuritic plaques, diffuse plaques, and neurofibrillary tangles were visualized with Bielschowsky silver stain, and counted and summarized to yield a Braak stage, Consortium to Establish a Registry for Alzheimer's Disease (CERAD) diagnosis, National Institute on Aging (NIA) Reagan diagnosis, and composite measure of AD pathology. The authors recorded the number and location of all gross chronic cerebral infarctions. Lewy bodies were identified with antibodies to alpha-synuclein. Multiple regression analyses were used to examine the relation of AD pathology and cerebral infarctions to clinical diagnosis proximate to death, controlling for age, sex, and education. RESULTS: A total of 37 had mild cognitive impairment, 60 did not have cognitive impairment, and 83 had dementia proximate to death. Nearly all persons had at least some AD pathology. Cerebral infarctions were present in 35.2%, and 15.6% had Lewy body disease. Persons with mild cognitive impairment were intermediate in terms of Braak stage and CERAD and NIA-Reagan neuropathologic criteria for AD compared to the other two groups. In multiple regression analyses, persons with mild cognitive impairment had intermediate levels of AD pathology from those without cognitive impairment and those with dementia (test for trend, F = 45.2, p < 0.001). Further, the relation between cognition and AD pathology in persons with mild cognitive impairment did not differ significantly from the relation between cognition and AD pathology in persons with dementia or those without cognitive impairment. Persons with mild cognitive impairment also had intermediate levels of cerebral infarctions (test for trend, p = 0.04). Only 3 (8.1%) persons with mild cognitive impairment had Lewy body disease. CONCLUSION: These data suggest that mild cognitive impairment may be the earliest clinical manifestation of common age-related neurologic diseases. PMID- 15753420 TI - Markers of immune activation and viral load in HIV-associated sensory neuropathy. AB - BACKGROUND: HIV infection is associated with a painful distal sensory polyneuropathy (DSP) that can severely limit the quality of life of affected subjects. The pathogenesis of DSP is unknown, although both HIV proteins and products of immune activation triggered by HIV infection have been implicated. OBJECTIVE: To assess the association between baseline markers of immune activation and HIV RNA levels (viral load) and time to symptomatic DSP (SDSP). METHODS: A cohort of 376 subjects, most receiving highly active antiretroviral therapy (HAART), were followed semiannually for up to 48 months. Blood and CSF levels of HIV viral load, monocyte chemotactic protein-1, macrophage colony stimulating factor (M-CSF), matrix metalloproteinase-2, and tumor necrosis factor alpha were measured in addition to CD4 lymphocyte cell count. RESULTS: In subjects without SDSP at baseline (62.5% of the cohort), among the virologic and immunologic markers, only baseline CSF M-CSF levels were associated with time to SDSP (hazard ratio = 2.97, p = 0.05). The Kaplan-Meier estimate of the 1-year incidence of SDSP was 21%, a 15% decrease from that observed in the Dana cohort, a pre-HAART cohort enrolled with the same inclusion/exclusion criteria. CONCLUSION: Highly active retroviral therapy (HAART) has changed the natural history of HIV-associated symptomatic distal sensory polyneuropathy (SDSP), which may explain, in contrast with studies from the pre-HAART era, the lack of association between SDSP and baseline HIV viral load and CD4 cell count. PMID- 15753421 TI - Radiologic evidence for absence of the facial nerve in Mobius syndrome. AB - OBJECTIVE: To detail the radiologic findings in Mobius syndrome, in order to clarify its pathogenetic mechanisms. METHODS: High resolution three-dimensional T1 (MP rage) and T2 (CISS) weighted MRI were used to study the cisternal and canalicular portion of the seventh cranial nerve in six Mobius patients. Also, the anteroposterior dimension of the brainstem was measured at the level of the pons in the authors' 6 patients and in 20 age-matched healthy control subjects. Furthermore, the MRIs were evaluated for associated congenital brain anomalies. RESULTS: The facial nerves were absent in all six patients despite residual function in some facial muscles. The authors confirmed brainstem hypoplasia but did not find tegmental calcifications. The anteroposterior dimension of the brainstem ranged between 17 and 25 mm vs 20 to 27 mm for controls. In three patients there were congenital abnormalities in the posterior fossa. CONCLUSION: The absent facial nerves on MRI and the unusual distribution of the facial weakness, which is characteristic of Mobius syndrome, suggests that other cranial nerves, possibly the trigeminal, hypoglossal, or glossopharyngeal nerve, aberrantly innervate some lower facial muscles. Radiologic findings support the notion that Mobius syndrome is part of a more complex congenital anomaly of the fossa posterior. PMID- 15753422 TI - Patterns and serial changes in electrodiagnostic abnormalities of axonal Guillain Barre syndrome. AB - BACKGROUND: In Guillain-Barre syndrome (GBS), anti-ganglioside antibodies are strongly associated with the acute motor axonal neuropathy (AMAN) form, but there are also cases of the demyelinating form of GBS (acute inflammatory demyelinating polyneuropathy [AIDP]) with anti-ganglioside antibodies. OBJECTIVE: To elucidate the patterns and sequential changes in electrodiagnostic abnormalities of anti ganglioside-positive GBS. METHODS: Detailed serial electrodiagnostic findings were reviewed for 51 patients with GBS. Anti-ganglioside antibodies were measured by ELISA. RESULTS: Antibodies to GM1, GM1b, GD1a, or GalNAc-GD1a were present in 25 patients. Of these, 12 (48%) showed the AMAN pattern, 5 (20%) the AIDP pattern, and 3 (12%) isolated F-wave absence in the first examination. All five patients with the AIDP pattern showed prolonged distal latencies, but three eventually showed the AMAN pattern or rapid normalization. The remaining two still had similarly prolonged distal latencies in weeks 4 to 6, but the serial changes were distinct from those in the anti-ganglioside-negative AIDP patients who showed progressive increases in distal latencies over 2 months after onset. CONCLUSIONS: Besides the simple axonal degeneration pattern, patients with anti ganglioside-positive Guillain-Barre syndrome can show transient conduction slowing/block in the distal or proximal nerve segments, mimicking demyelination, but anti-ganglioside antibodies do not appear to be associated with acute inflammatory demyelinating polyneuropathy. PMID- 15753423 TI - Temporal lobe atrophy on MRI in Parkinson disease with dementia: a comparison with Alzheimer disease and dementia with Lewy bodies. AB - OBJECTIVE: To investigate the extent of medial temporal lobe atrophy (MTA) on MRI in Parkinson disease (PD) with and without dementia compared with Alzheimer disease (AD) and dementia with Lewy bodies (DLB) and to determine whether MTA correlates with cognitive impairment in PD and PD dementia (PDD). METHODS: Coronal T1-weighted MRI scans were acquired from control subjects (n = 39) and patients with PD (n = 33), PDD (n = 31), DLB (n = 25), and AD (n = 31), diagnosed according to standardized clinical diagnostic criteria. Cognitive function was assessed using the Cambridge Cognitive Examination (CAMCOG), and MTA was rated visually using a standardized (Scheltens) scale. RESULTS: More severe MTA was seen in PDD (p = 0.007), DLB (p < 0.001), and AD (p < 0.001) vs control subjects. PD subjects had greater hippocampal atrophy than control subjects (p = 0.015) but less than subjects with DLB and AD, though not with PDD. MTA correlated with CAMCOG score and memory scores in the DLB group and with age in control, PDD, and AD groups. There were no correlations between MTA and cognitive impairment in PD, PDD, and AD. PDD and DLB had a similar profile of cognitive impairment and MTA. CONCLUSIONS: Medial temporal lobe atrophy (MTA) was seen in cognitively intact older subjects with Parkinson disease (PD) and was not more pronounced in Parkinson disease dementia (PDD). Alzheimer disease (AD) and, to a lesser extent, dementia with Lewy bodies (DLB) showed more pronounced MTA. Results suggest early hippocampal involvement in PD and that when dementia develops in PD, anatomic structures apart from the hippocampus are predominantly implicated. Greater hippocampal involvement in AD vs PDD and DLB is consistent with clinical, cognitive, and pathologic differences between the disorders. PMID- 15753424 TI - Endogenous sex hormone levels and cognitive function in aging men: is there an optimal level? AB - OBJECTIVE: To determine whether endogenous sex hormone levels are associated with cognitive functioning in men. METHODS: Cognitive performance was assessed in 400 independently living men between ages 40 and 80 in a population-based cross sectional study. Compound scores were calculated for memory function, processing capacity/speed, and executive function. The Mini-Mental State Examination was used as a measure of global cognitive function. The adjusted association of testosterone (T) and estradiol (E2) (total, bioavailable) with neuropsychological test scores in the total group and in subgroups was assessed by linear and logistic regression analysis. RESULTS: Curvilinear associations were observed between T and memory performance and processing capacity/speed, suggesting optimal sex hormone levels. No association between E2 and cognitive functioning was found. After the population was subdivided into four age decades, a linear association of T with cognitive functioning in the oldest age category remained. No association was found in the other age decades. Lower bioavailable T levels were associated with lower scores on processing capacity/speed and executive function; beta (95% CI) values were 0.36 (0.07 to 0.66) and 0.17 (-0.01 to 0.35). Similar results were observed for total T. CONCLUSIONS: Higher testosterone (T) levels are associated with better cognitive performance in the oldest age category. Apparent curvilinear associations between T and certain cognitive functions in men suggest an optimal hormone level for particular cognitive tasks and are explained by linear associations in the oldest age category. PMID- 15753425 TI - Safety and cognitive effect of frontal DC brain polarization in healthy individuals. AB - BACKGROUND: Data from the human motor cortex suggest that, depending on polarity, direct current (DC) brain polarization can depress or activate cortical neurons. Activating effects on the frontal lobe might be beneficial for patients with frontal lobe disorders. This phase 1 study tested the safety of frontal DC, including its effects on frontal and other brain functions. METHODS: The authors applied 20 minutes of anodal, cathodal, or sham DC to the left prefrontal cortex in three groups of right-handed subjects and looked for effects on global measures of processing and psychomotor speed, emotion, and verbal fluency, a measure of local cortical function. In one experiment (n = 30), the authors tested before and after 1 mA DC and monitored EEG in 9 subjects. In two other experiments using 1 mA (n = 43) and 2 mA (n = 30), the authors tested before and then starting 5 minutes after the onset of DC. RESULTS: All subjects tolerated DC well. There were no significant effects on performance with 1 mA DC. At 2 mA, verbal fluency improved significantly with anodal and decreased mildly with cathodal DC. There were no clinically significant effects on the other measures. CONCLUSIONS: Limited exposure to direct current polarization of the prefrontal cortex is safe and can enhance verbal fluency selectively in healthy subjects. As such, it deserves consideration as a procedure to improve frontal lobe function in patients. PMID- 15753426 TI - Midazolam in neonatal seizures with no response to phenobarbital. AB - The outcome of 45 neonates with EEG-confirmed seizures (ESz) was analyzed with regard to treatment. ESz persisted in 17 of 32 neonates receiving phenobarbital/phenytoin (13 had a poor outcome, 4 died). In contrast, ESz were rapidly controlled in 13 of 13 nonresponders to phenobarbital/phenytoin treated with midazolam (4 had poor outcome, 2 died). Nonresponders to phenobarbital/phenytoin had a significantly worse outcome than responders did. Midazolam effectively controlled ESz in nonresponders to phenobarbital/phenytoin and correlated with significantly improved long-term neurodevelopment. PMID- 15753427 TI - Mortality following a first unprovoked seizure in children: a prospective study. AB - In a prospective study, 407 children with a first unprovoked seizure were followed for a mean of 14.2 years. To date, nine have died. Death was unrelated to seizures in four subjects who had no further seizures and were on no medications. The remaining five subjects all had multiple seizures and were on medications. Treatment following the first seizure would not have altered mortality in any of the nine cases. PMID- 15753428 TI - REM behavior disorder associated with epileptic seizures. AB - Reported is the association of REM behavior disorder (RBD) with late-onset, sleep related, tonic-clonic seizures in two elderly men. In each patient, RBD preceded the onset of seizures by several years. The authors hypothesize that REM sleep disruption may facilitate seizure occurrence. PMID- 15753429 TI - MRI brain volumetry in Rasmussen encephalitis: the fate of affected and "unaffected" hemispheres. AB - Rasmussen encephalitis (RE) is regarded as a unihemispheric disease. Serial three dimensional MRIs of 18 patients under immunotherapy were analyzed volumetrically and planimetrically. Median volume loss was significantly higher in the affected than in the unaffected hemispheres (29.9 cm3/y vs 6.8 cm3/y). Correlation of the planimetrically and volumetrically assessed hemispheric ratios (HRs) was significant. The results support the concept of RE as a basically unilateral disease. Planimetric HR assessment is valid and time efficient. PMID- 15753430 TI - Treatment of early onset multiple sclerosis with subcutaneous interferon beta-1a. AB - The authors studied the tolerability of subcutaneous interferon beta-1a (IFNbeta 1a) in 51 patients with early-onset multiple sclerosis. The most frequent systemic adverse effects were flu-like symptoms in 65%. Laboratory abnormalities included asymptomatic leukopenia (27%) and elevated hepatic transaminases (35%). Treatment with IFNbeta-1a was safe and well tolerated in the majority of children and adolescents. PMID- 15753431 TI - The cognitive burden of multiple sclerosis in children. AB - Little is known about the cognitive sequelae of pediatric-onset multiple sclerosis (MS). Ten pediatric patients with MS were evaluated using a comprehensive neuropsychological battery. Neuropsychological deficits were identified on measures of general cognition, language, visuomotor integration, and verbal and visual memory. Cognitive impairment occurs in children with MS, and those with longer disease duration and younger age at MS onset appear to be at greatest risk. PMID- 15753432 TI - Alzheimer disease and cancer. AB - Cross-sectional studies raise the possibility of protective relationships between, or a common mechanism underlying, the development of dementia of the Alzheimer type (DAT) and cancer. Using a prospective longitudinal design, the authors found that the risk of developing cancer is less among participants with DAT vs nondemented participants (p < 0.001) and that the risk of developing DAT may be less for participants with a history of cancer (p = 0.060). PMID- 15753433 TI - Hypertension and cognitive performance in African Americans with Alzheimer disease. AB - The authors examined the relationship between hypertension and cognitive performance in 34 African-American patients with probable Alzheimer disease. Multiple regression analyses indicated that hypertension was associated with poorer overall performance on the Mattis Dementia Rating Scale, particularly the Initiation/Perseveration and Conceptualization subscales, after controlling for gender, age, and education. The findings suggest that African-American patients with hypertension exhibit greater cognitive impairment, possibly reflecting executive dysfunction. PMID- 15753434 TI - DWI predicts future progression to Alzheimer disease in amnestic mild cognitive impairment. AB - The authors assessed whether measures of hippocampal water diffusivity at baseline can predict future progression to Alzheimer disease (AD) in amnestic mild cognitive impairment (aMCI). Higher baseline hippocampal diffusivity was associated with a greater risk of progression to AD in aMCI (p = 0.002). Magnetic resonance diffusion-weighted imaging may help identify patients with aMCI who will progress to AD as well as or better than structural MRI measures of hippocampal atrophy. PMID- 15753435 TI - Creutzfeldt-Jakob disease associated with the R208H mutation in the prion protein gene. AB - The authors investigated a patient who died of apparent sporadic Creutzfeldt Jakob disease (CJD) but carried a R208H substitution in the prion protein (PrP). The patient phenotype was indistinguishable from typical sporadic CJD (i.e., MM1 subtype). In addition, pathologic PrP, PrP(Sc), originated from both the normal and the mutated PRNP allele and had the same characteristics as PrP(Sc) type 1. The authors propose that the R208H mutation influences disease susceptibility without significantly affecting PrP(Sc) properties or disease phenotype. PMID- 15753436 TI - High mutation rate in dopa-responsive dystonia: detection with comprehensive GCHI screening. AB - Mutations in GTP cyclohydrolase I (GCHI) are found in 50 to 60% of cases with dopa-responsive dystonia (DRD). Heterozygous GCHI exon deletions, undetectable by sequencing, have recently been described in three DRD families. We tested 23 individuals with DRD for the different mutation types by conventional and quantitative PCR analyses and found mutations, including two large exon deletions, in 87%. The authors attribute this high mutation rate to rigorous inclusion criteria and comprehensive mutational analysis. PMID- 15753437 TI - MTX-induced white matter changes are associated with polymorphisms of methionine metabolism. AB - Methotrexate (MTX) is a folate antagonist inhibiting nucleic acid and methionine synthesis. Methionine is necessary for CNS myelination. In 42 patients with primary CNS lymphoma (PCNSL) treated with a systemic and intraventricular high dose MTX-based polychemotherapy, the presence of a risk haplotype defined by polymorphisms influencing methionine metabolism referred a relative risk for CNS white matter changes of 4.7 (p = 0.001). The authors conclude that methionine metabolism influences MTX neurotoxicity. PMID- 15753439 TI - Medical use of cannabis in the Netherlands. AB - The authors investigated the indications for cannabis prescription in the Netherlands and assessed its efficacy and side effects. A majority (64.1%) of patients reported a good or excellent effect on their symptoms. Of these patients, approximately 44% used cannabis for >/=5 months. Indications were neurologic disorders, pain, musculoskeletal disorders, and cancer anorexia/cachexia. Inhaled cannabis was perceived as more effective than oral administration. Reported side effects were generally mild. PMID- 15753438 TI - Chronic inflammatory demyelinating polyradiculoneuropathy: MRI study of brain and spinal cord. AB - CNS demyelinating lesions have been reported in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). There are no studies of cord atrophy in CIDP. Ten patients with CIDP underwent brain and spinal cord MRI to investigate CNS demyelination and cord atrophy. No CNS demyelination was found, but the mean cervical cord area was significantly smaller in CIDP patients vs control subjects. Spinal cord atrophy may be related to degeneration secondary to axonal loss. PMID- 15753440 TI - Intracranial fat embolization due to baclofen pump. PMID- 15753441 TI - Natural course of the remission of vertigo in patients with benign paroxysmal positional vertigo. AB - The authors assessed the natural course of benign paroxysmal positional vertigo (BPPV) in 108 patients who were not treated with canalith repositioning procedure. The average number of days from onset to remission of positional vertigo in patients with posterior canal BPPV (P-BPPV) (39 days) was longer than in those with horizontal canal BPPV (H-BPPV) (16 days). The ratio of patients with H-BPPV to those with BPPV was 33%. PMID- 15753443 TI - "Levodopa phobia": a new iatrogenic cause of disability in Parkinson disease. PMID- 15753442 TI - An unusual cause of dysphagia and dysphonia. PMID- 15753444 TI - Aneurysmal subarachnoid leak with normal CT and CSF spectrophotometry. PMID- 15753445 TI - Palatal myoclonus in a patient with a lateral thalamic infarction. PMID- 15753446 TI - Different pain patterns in patients with vertebral artery dissections. PMID- 15753447 TI - Severe sensory neuropathy associated with long-term linezolid use. PMID- 15753448 TI - Electrophysiology extends the phenotypic spectrum of X-linked myopathy with excessive autophagy. PMID- 15753449 TI - Fiction: the Christmas card. PMID- 15753450 TI - Early neurologic deterioration in intracerebral hemorrhage: predictors and associated factors. PMID- 15753451 TI - A randomized trial of frovatriptan for the intermittent prevention of menstrual migraine. PMID- 15753452 TI - Progressive dementia after first-ever stroke: a community-based follow-up study. PMID- 15753453 TI - No evidence for heritability of Parkinson disease in Swedish twins. PMID- 15753454 TI - Multiple sclerosis and occult gluten sensitivity. PMID- 15753455 TI - Salvage therapy for primary CNS lymphoma with a combination of rituximab and temozolomide. PMID- 15753456 TI - Critical update and emerging trends in epidermal growth factor receptor targeting in cancer. AB - The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase of the ErbB receptor family that is abnormally activated in many epithelial tumors. The aberrant activation of the EGFR leads to enhanced proliferation and other tumor promoting activities, which provide a strong rationale to target this receptor family. There are two classes of anti-EGFR agents: monoclonal antibodies (MAbs) directed at the extracellular domain of the receptor and small molecule, adenosine triphosphate-competitive inhibitors of the receptor's tyrosine kinase. Anti-EGFR MAbs have shown antitumor activity in advanced colorectal carcinoma, squamous cell carcinomas of the head and neck, non-small-cell lung cancer (NSCLC) and renal cell carcinomas. The tyrosine kinase inhibitors (TKIs) have a partially different activity profile. They are active against NSCLC, and a specific EGFR inhibitor has shown improvement in survival. Recently, mutations and amplifications of the EGFR gene have been identified in NSCLC and predict for enhanced sensitivity to anti-EGFR TKIs. In addition to specific anti-EGFR TKIs, there are broader acting inhibitors such as dual EGFR HER-2 inhibitors and combined anti-pan-ErbB and antivascular endothelial growth factor receptor inhibitors. Current research efforts are directed at selecting the optimal dose and schedule and identifying predictive factors of response and resistance beyond EGFR gene mutations and/or amplifications. Finally, there is a need for improved strategies to integrate anti-EGFR agents with conventional therapies and to explore combinations with other molecular targeted approaches including other antireceptor therapies, receptor-downstream signaling transduction inhibitors, and targeted approaches interfering with other essential drivers of cancer, such as angiogenesis. PMID- 15753457 TI - Promoter hypermethylation: a new therapeutic target emerges in urothelial cancer. PMID- 15753458 TI - Full haplotype-mismatched hematopoietic stem-cell transplantation: a phase II study in patients with acute leukemia at high risk of relapse. AB - PURPOSE: Establishment of hematopoietic stem-cell (HSC) transplantation from mismatched relatives is feasible for patients with acute leukemia. As our original method of graft processing was unsuitable for large-scale clinical studies, we use automated devices for CD34+ cell purification. PATIENTS AND METHODS: Sixty-seven patients with acute myeloid leukemia (AML; 19 complete remission [CR] 1, 14 CR 2, nine CR > 2, 25 in relapse) and 37 with acute lymphoid leukemia (ALL; 14 CR 1, eight CR 2, two CR > 2, 13 in relapse) were conditioned with total-body irradiation, thiotepa, fludarabine, and antithymocyte globulin. Peripheral-blood progenitor cells were mobilized with recombinant human granulocyte colony-stimulating factor and depleted of T-cells using CD34+ cell immunoselection. No post-transplantation graft-versus-host disease (GvHD) prophylaxis was administered. RESULTS: Primary engraftment was achieved in 94 of 101 assessable patients. Six of the seven patients who rejected the primary graft, engrafted after a second transplantation. Overall, 100 of 101 patients engrafted. Acute GvHD developed in eight of 100 patients, and chronic GvHD, in five of 70 assessable patients. Thirty-eight patients died of nonleukemic causes. Relapse occurred in nine of 66 patients receiving transplantation in remission and in 17 of 38 receiving transplantation in relapse. Median follow-up of the 40 patients who survived event-free was 22 months (range, 1 to 65 months). Event free survival (+/- standard deviation) rate was 48% +/- 8% and 46% +/- 10%, respectively, for the 42 AML and 24 ALL patients receiving transplantation in remission. CONCLUSION: Our transplantation procedure provides reliable, reproducible CD34+ cell purification, high engraftment rates, and prevention of GvHD. The mismatched-related transplant emerges as a viable, alternative source of stem cells for acute leukemia patients without matched donors and/or those who urgently need transplantation. PMID- 15753459 TI - Evaluation of a 7-day continuous intravenous infusion of decitabine: inhibition of promoter-specific and global genomic DNA methylation. AB - PURPOSE: The nucleoside analog 5-aza-2'-deoxycytidine (5-aza-CdR, decitabine) is a potent inhibitor of DNA methylation in vitro. Cellular treatment with this agent induces the re-expression of methylation-silenced genes. It remains unclear to what extent this compound inhibits DNA methylation in vivo. A clinical study was designed to examine the molecular effects and toxicity of a continuous 1-week intravenous infusion of decitabine in solid tumor patients. METHODS: Ten patients with refractory solid tumors were included in this study. Decitabine was administered at 2 mg/m(2)/d [DOSAGE ERROR CORRECTED] via continuous infusion for 168 hours. Quantitative polymerase chain reaction and high performance liquid chromatography were utilized to measure promoter-specific and global DNA methylation in peripheral-blood cells before and after treatment. RESULTS: Transient grade III/IV neutropenia (two patients) and grade II thrombocytopenia (one patient) was observed at the lowest planned dose step (2 mg/m2/d for 7 days). Nonhematologic toxicities were not observed. Quantitative polymerase chain reaction demonstrated significant MAGE-1 promoter hypomethylation by 14 days after the start of treatment in all 13 treatment cycles examined. Significant genomic DNA hypomethylation was also seen by day 14 in 11 of 13 treatment cycles analyzed. Genomic DNA methylation reverted to baseline levels by 28 to 35 days after the start of treatment, demonstrating that inhibition of DNA methylation by decitabine is transient. CONCLUSION: A 168-hour continuous infusion of decitabine is well tolerated and results in the inhibition of promoter-specific and genomic DNA methylation in vivo. This treatment schedule is suitable for evaluation of decitabine in combination with agents whose activity may be enhanced by the reversal of DNA methylation-mediated gene silencing. PMID- 15753460 TI - Increasing incidence of late second malignancies after conditioning with cyclophosphamide and total-body irradiation and autologous bone marrow transplantation for non-Hodgkin's lymphoma. AB - PURPOSE: Although the risk of myelodysplastic syndrome (MDS) has been well described following autologous bone marrow transplantation (ABMT), the risk of solid tumors has been poorly characterized. We report the incidence and outcome of solid tumors at 10-year follow-up in a large cohort of uniformly treated patients who underwent ABMT for non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Between 1982 and 1997, 605 patients underwent ABMT for B-cell NHL, with uniform conditioning with cyclophosphamide and total-body irradiation followed by reinfusion of autologous bone marrow purged with anti-B-cell monoclonal antibodies. Current information on relapse of disease and second malignancies was obtained via an institutional review board-approved questionnaire sent to the referring oncologists. RESULTS: Forty-two solid tumors, six non-MDS hematologic malignancies, 39 nonmelanoma skin cancers, and 68 cases of MDS/acute myelogenous leukemia (AML) were observed at a median follow-up of 9.5 years. A cumulative incidence model using death as a competing risk found that the 10-year incidence of second malignancy is 21%, with 10.0% non-MDS malignancies. The projected incidence of all malignancies at 15 years is 29%. The principal risk factor for second malignancy is increased age at ABMT (P = .0002). In the entire cohort, 9.6% of patients have died of second malignancy. CONCLUSION: Lengthy follow-up demonstrates a significant incidence of second malignancies after ABMT for NHL. Although the incidence of MDS/AML starts to plateau, the incidence of solid tumors continues to rise. Second malignancies are responsible for a significant fraction of overall mortality following ABMT. PMID- 15753461 TI - Promoter hypermethylation is associated with tumor location, stage, and subsequent progression in transitional cell carcinoma. AB - PURPOSE: Transitional cell carcinoma (TCC) is a pan-urothelial disease characterized by multiplicity. Although little is known about the molecular events in upper-tract TCC, similar carcinogenic mechanisms are thought to occur throughout the urinary tract. However, we have previously shown that distinct patterns of microsatellite instability occur in upper and lower urinary tract TCC, suggesting biologic differences between these tumors. Here we investigate the extent of promoter hypermethylation in TCC throughout the urinary tract. PATIENTS AND METHODS: Tissue was obtained from 280 patients (median follow-up, 56 months) whose tumors comprised 116 bladder and 164 upper-tract tumors (UTT). Analysis for hypermethylation at 11 CpG islands, using methylation-sensitive polymerase chain reaction and bisulfite sequencing, was performed for each sample and compared with the tumor's clinicopathologic details, microsatellite instability status, and subsequent behavior. RESULTS: Promoter methylation was present in 86% of TCC and occurred both more frequently and more extensively in UTT (94%) than in bladder tumors (76%; P < .0001). Methylation was associated with advanced tumor stage (P = .0001) and higher tumor progression (P = .03) and mortality rates (P = .04), when compared with tumors without methylation. Multivariate analysis revealed that methylation at the RASSF1A and DAPK loci, in addition to tumor stage and grade, were associated with disease progression (P < .04). CONCLUSION: Despite morphologic similarities, there are genetic and epigenetic differences between TCC in the upper and lower urinary tracts. Methylation occurs commonly in urinary tract tumors, may affect carcinogenic mechanisms, and is a prognostic marker and a potential therapeutic target. PMID- 15753462 TI - Phase I/II trial evaluating the anti-vascular endothelial growth factor monoclonal antibody bevacizumab in combination with the HER-1/epidermal growth factor receptor tyrosine kinase inhibitor erlotinib for patients with recurrent non-small-cell lung cancer. AB - PURPOSE: Bevacizumab (Avastin; Genentech, South San Francisco, CA) is a recombinant, humanized anti-vascular endothelial growth factor monoclonal antibody. Erlotinib HCl (Tarceva, OSI-774; OSI Pharmaceuticals, New York, NY) is a potent, reversible, highly selective and orally available HER-1/epidermal growth factor receptor tyrosine kinase inhibitor. Preclinical data in various xenograft models produced greater growth inhibition than with either agent alone. Additionally, both agents have demonstrated benefit in patients with previously treated non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: A phase I/II study in two centers examined erlotinib and bevacizumab (A+T) in patients with nonsquamous stage IIIB/IV NSCLC with > or = one prior chemotherapy. In phase I, erlotinib 150 mg/day orally plus bevacizumab 15 mg/kg intravenously every 21 days was established as the phase II dose, although no dose-limiting toxicities were observed. Phase II assessed the efficacy and tolerability of A+T at this dose. Pharmacokinetic parameters were evaluated. ResultsForty patients were enrolled and treated in this study (34 patients at phase II dose); the median age was 59 years (range, 36 to 72 years), 21 were female, 30 had adenocarcinoma histology, nine were never-smokers, and 22 had > or = two prior regimens (three patients had > or = four prior regimens). The most common adverse events were mild to moderate rash, diarrhea, and proteinuria. Preliminary data showed no pharmacokinetic interaction between A + T. Eight patients (20.0%; 95% CI, 7.6% to 32.4%) had partial responses and 26 (65.0%; 95% CI, 50.2% to 79.8%) had stable disease as their best response. The median overall survival for the 34 patients treated at the phase II dose was 12.6 months, with progression-free survival of 6.2 months. CONCLUSION: Encouraging antitumor activity and safety of A + T support further development of this combination for patients with advanced NSCLC and other solid tumors. PMID- 15753463 TI - Molecular changes in tamoxifen-resistant breast cancer: relationship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. AB - PURPOSE: To evaluate growth factor receptor cross talk with the estrogen receptor (ER) in paired clinical breast cancer specimens and in a xenograft model before tamoxifen and at tumor progression as a possible mechanism for tamoxifen resistance. METHODS: Specimen pairs from 39 patients were tissue arrayed and stained for ER, progesterone receptor (PgR), Bcl-2, c-ErbB2 (HER-2), and phosphorylated (p) p38 mitogen-activated protein kinase (MAPK), p-ERK1/2 MAPK, and p-Akt. Xenograft MCF-7 tumors before and after tamoxifen resistance were assessed for levels of p-p38. RESULTS: Pretreatment, there were strong correlations between ER, PgR, and Bcl-2, and an inverse correlation between ER and HER-2. These correlations were lost in the tamoxifen- resistant tumors and replaced by strong correlations between ER and p-p38 and p-ERK. ER expression was lost in 17% of resistant tumors. Three (11%) of the 26 tumors originally negative for HER-2 became amplified and/or overexpressed at resistance. All ER-positive tumors that overexpressed HER-2 originally or at resistance expressed high levels of p-p38. In the pretreatment and tamoxifen-resistant specimens, there were strong correlations between p-p38 and p-ERK. In the tamoxifen-resistant xenograft tumors, like the clinical samples, there was a striking increase in p-p38. CONCLUSION: The molecular pathways driving tumor growth can change as the tumor progresses. Crosstalk between ER, HER-2, p38, and ERK may contribute to tamoxifen resistance and may provide molecular targets to overcome this resistance. PMID- 15753464 TI - Systemic AA amyloidosis in the common marmoset. AB - The common marmoset (Callithrix jacchus) is a small New World primate native to Brazil that has been used extensively in biomedical research. A retrospective analysis of archived hematoxylin and eosin-stained tissue sections and clinical records was conducted at the New England Primate Research Center on 86 marmosets more than 1 year of age that were euthanized during the past decade because of morbidity and failure to thrive. Approximately 17% (15 of 86) were found to have amyloid deposits in one or more organs, including the liver, adrenal glands, kidneys, and intestine. This material was shown by amino acid sequence analysis to be composed of serum amyloid A (SAA)-related protein. This type of amyloidosis, designated AA or "secondary," is associated typically with an inflammatory process that induces elevated levels of the SAA amyloidogenic precursor molecule. Notably, there were no significant pathologic differences or other distinguishing features in animals with amyloid versus those without; furthermore, on the basis of the limited number of serum specimens available for analysis, the SAA concentrations in the two groups were comparable, thus suggesting the possible inheritable nature of the disorder. In this respect, the common marmoset provides a unique experimental model for study of the pathogenesis and treatment of AA and other forms of systemic amyloidosis. PMID- 15753465 TI - Bacterial diskospondylitis associated with posterior paresis/paralysis in North American farmed mink (Mustela vison). AB - Posterior paresis/paralysis in farmed mink is responsible for significant morbidity and mortality, with individual farms reporting the loss of as many as 700 animals each year. Although this disease has been recognized by North American mink farmers for approximately 40 years, there are few published reports focusing on this entity. The objective of this study was to investigate the etiology and pathogenesis of the disease. Complete necropsy examinations were done on 40 clinically affected mink, ranging from 7 to 10 weeks of age, and on three normal animals in the same age range from two mink farms. Thirty-two of the 40 clinically affected animals had an isolated vertebral lesion characterized by bone lysis and proliferation that usually was centered on an intervertebral disk space in the midthoracic area. An inflammatory reaction, composed primarily of neutrophils, was present within the vertebral sections in 25 of the 40 affected animals (62.5%), and the presence of gram-positive cocci was confirmed in 8 of 10 animals (80%) in which bacterial organisms were observed histologically. Bacterial cultures from 15 affected animals yielded Streptococcus sp. from the intervertebral disk space in 13 of 15 (86.7%) animals and from heart blood in 6 of 8 (75%). A farm visit revealed no history or evidence of traumatic wounds as a source of infection in these animals, and the diet appeared to be adequate for skeletal development. We conclude that posterior paresis/paralysis in farmed mink is associated with bacterial diskospondylitis, likely occurring secondary to bacteremia/septicemia. PMID- 15753466 TI - An ancillary tool for the diagnosis of amyloid A amyloidosis in a variety of domestic and wild animals. AB - Immunohistochemistry, the standard method for diagnosing amyloid A (AA) amyloidosis, is limited in animals because it requires a large array of animal specific anti-AA antibodies, not commercially available. The Shtrasburg method (SH method) is a highly specific and sensitive technique, helping in the diagnosis and determination of AA amyloidosis in humans. The aim of this study is to determine whether the SH method is applicable in the diagnosis of AA amyloidosis in a variety of animals. Tissue samples were obtained from animals suffering from spontaneous or experimentally induced AA amyloidosis (mice, hamsters, guinea pigs, cheetahs, cats, cows, ducks, a dog, a goose, a chicken, and a turaco). Detection of the amyloid and quantitative evaluation were performed using Congo red staining, and specific AA typing was performed by the potassium permanganate technique. The studied tissues were subjected to the SH method, which confirmed the AA nature of the amyloid deposit, by displaying in polyacrylamide gel electrophoresis protein bands consistent with the molecular weight of the species-specific AA, in all the animals examined, except mice, hamsters, and guinea pigs. N-terminal analysis of these bands corroborated their AA origin. We conclude that the SH method may be used as an ancillary simple tool for the diagnosis of AA amyloidosis in a large number of domestic and wild animals. Moreover, our findings further increase the feasibility of applying this method in humans. PMID- 15753467 TI - Canine hepatic neuroendocrine carcinoma: an immunohistochemical and electron microscopic study. AB - Ten dogs with neuroendocrine carcinoma of the liver were selected for inclusion in the study. Clinical signs were anorexia (7), vomiting (5), polydipsia/polyuria (3), icterus (2), lethargy (2), weight loss (2), paresis (1), ataxia (1), weakness (1), collapse (1), and urinary tract infection (1). Hematologic and biochemical abnormalities included anemia (2/8), leukocytosis (4/8), high liver enzyme activity (serum alkaline phosphatase, 7/9; alanine transaminase, 7/9; aspartate transaminase, 8/9), and high total bilirubin (6/9). Grossly, the tumors were diffuse, involving all liver lobes in six dogs, and two dogs had various sized nodules in addition to diffuse involvement. Histologically, there were eight tumors with solid or trabecular pattern (group A), one tumor with cords or rows of neoplastic cells (group B), and one tumor with multiple rosette-like structures (group C). Immunohistochemical studies revealed that all 10 neoplasms were positive for at least one of the endocrine markers used: neuron-specific enolase (NSE; 8/10), synaptophysin (5/10), and chromogranin-A (3/10). A panel of NSE, chromagranin-A, and synaptophysin detected 100% of the tumors in our series. Electron microscopy confirmed the diagnosis by the presence of intracytoplasmic neurosecretory granules in the two examined cases. Our results show that neuroendocrine markers commonly used in humans can be used for the diagnosis of hepatic neuroendocrine carcinoma in dogs, preferably a panel of synaptophysin, chromagranin-A, and NSE because chromogranin-A alone is not as useful in dogs as in humans. PMID- 15753468 TI - Liver histopathology and liver and serum alanine aminotransferase and alkaline phosphatase activities in epileptic dogs receiving phenobarbital. AB - Phenobarbital (PB) therapy is frequently associated with elevated serum alanine aminotransferase (ALT) and alkaline phosphatase (AP) activities in dogs without clinical signs of liver disease. The goal of this study was to determine if increased serum ALT and AP activities in clinically healthy PB-treated epileptic dogs are due to hepatic enzyme induction or to subclinical liver injury. Liver biopsies were obtained from 12 PB-treated dogs without clinical signs of liver disease but with elevated serum ALT and/or AP activities or both. Liver biopsies were obtained from eight healthy control dogs not receiving PB. Biopsies were evaluated histopathologically (all dogs) and liver homogenates were assayed for ALT (all dogs) and AP (six treated dogs, all controls) activities. As a positive control, liver cytochrome P4502B, an enzyme known to be induced by PB, was measured by benzyloxyresorufin-O-dealkylase activity and immunoblotting (five treated dogs, all controls). Serum AP isoenzyme analyses were performed. Results showed that ALT and AP activities in liver homogenates were not increased in treated dogs compared with controls, whereas the positive control for induction, CYP2B, was dramatically increased in treated dogs. Histopathological examination of liver biopsies revealed more severe and frequent abnormalities in treated dogs compared to controls, but similar types of abnormalities were found in both groups. Serum AP isoenzyme analyses in treated dogs demonstrated increased corticosteroid-induced and liver isoenzyme activities compared to controls. Results do not support induction of ALT or AP in the liver as the cause of elevated serum activities of these enzymes due to PB. PMID- 15753469 TI - Use of alkaline phosphatase staining to differentiate canine osteosarcoma from other vimentin-positive tumors. AB - Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone neoplasia. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, and plasma cell myeloma. The purpose of this study is to determine the sensitivity and specificity of alkaline phosphatase (ALP) staining to differentiate OSA from other tumors that express vimentin by immunocytochemistry or immunohistochemistry. ALP is a hydrolytic enzyme present in multiple tissues including liver, kidney, intestine, placenta, and bone. Hypothetically, neoplasms actively producing bone should be specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 8-10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3 indolyl phosphate toluidine salt-phosphatase substrate. A positive reaction stains the membrane of the cells gray to black. Samples were counterstained with a Romanowsky's stain to determine whether the sample was of representative cellularity. A total of 61 vimentin-positive neoplasms have been evaluated and confirmed histopathologically. Tumors that expressed vimentin and were positive for ALP included 33 OSAs, one multi-lobular tumor of bone, one amelanotic melanoma, and one chondrosarcoma. Tumors that expressed vimentin and were negative for ALP included chondrosarcomas (three of four), multiple fibrosarcomas, and multiple synovial cell sarcomas. The sensitivity is 100%, and the specificity is 89%. In conclusion, ALP appears to be a highly sensitive and fairly specific marker in the diagnosis of OSA. PMID- 15753470 TI - Cutaneous leishmaniasis in naturally infected dogs is associated with a T helper 2-biased immune response. AB - Skin lesions are a frequent manifestation of Leishmania infantum infections in Mediterranean countries. This study demonstrates by real-time reverse transcriptase-polymerase chain reaction the local cytokine response in skin biopsies from Leishmania-infected dogs (n=10). As controls, we investigated skin biopsies from healthy (n=10) and fleabite hypersensitive dogs (n=10). We established a quantitative PCR to determine the parasite burden in biopsies. The objective was to elucidate whether a correlation exists between parasite number, histologic response, and T helper-1 (TH1)/T helper-2 (TH2) cytokine expression in lesional skin of naturally infected dogs. In Leishmania-infected dogs, interleukin-4 (IL-4), tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) messenger RNA production was significantly higher than controls. Furthermore, dogs with a high Leishmania burden had a significantly higher IL-4 expression, whereas no difference was noted with regard to expression of other cytokines. By comparing the pattern of inflammation and cytokine expression, a clear trend became evident in that levels of IL-4, TNF-alpha, and IFN-gamma were elevated in biopsies with a periadnexal nodular pattern and in biopsies where the severity of the periadnexal infiltrate was equal to the perivascular to interstitial infiltrate. Expression of IL-4, IL-13, and TNF-alpha was slightly increased in biopsies where plasma cells prevailed on lymphocytes, whereas expression of IFN-gamma was moderately higher when lymphocytes were predominating. In summary, the present study demonstrates that the local immune response in naturally occurring leishmaniasis includes TH1 as well as TH2 cytokine subsets. Furthermore, respective data suggest that increased expression of the TH2-type cytokine IL-4 is associated with both severe clinical signs and a high parasite burden in the skin lesions. PMID- 15753471 TI - Multiple perineuriomas in chicken (Gallus gallus domesticus). AB - Intraneural perineurioma is an extremely rare condition characterized by perineurial cell proliferation within peripheral nerve (PN) sheaths. In the veterinary field, this entity has been reported only in a dog. We examined multiple enlargements of PNs in 11 chickens (Gallus gallus domesticus) (9 Japanese bantams and 2 specific pathogen-free White Leghorn), which were inoculated with an avian leukosis virus (ALV) causing so-called fowl glioma. All chickens clinically exhibited progressive leg paralysis. Lumbosacral plexus, brachial plexus, and/or spinal ganglion were commonly affected, and these nerves contained a diffuse proliferation of spindle cells arranged concentrically in characteristic onion bulb-like structures surrounded by residual axons and myelin sheaths. The spindle cells were immunohistochemically negative for S 100alpha/beta protein. Electron microscopy revealed that these cells were characterized by short bipolar cytoplasmic processes, occasional cytoplasmic pinocytotic vesicles, and discontinuous basal laminae. These features are consistent with those of intraneural perineurioma. Furthermore, the specific sequence of the ALV was detected in the PN lesions of 8/11 (73%) birds by polymerase chain reaction. These results indicate that the multiple intraneural perineuriomas of chicken may be associated with the ALV-A causing fowl glioma. PMID- 15753472 TI - Pathology of domoic acid toxicity in California sea lions (Zalophus californianus). AB - Over 100 free-ranging adult California sea lions (Zalophus californianus) and one Northern fur seal (Callorhinus ursinus), predominantly adult females, were intoxicated by domoic acid (DA) during three harmful algal blooms between 1998 and 2000 in central and northern California coastal waters. The vector prey item was Northern anchovy (Engraulis mordax) and the primary DA-producing algal diatom was Psuedonitzschia australis. Postmortem examination revealed gross and histologic findings that were distinctive and aided in diagnosis. A total of 109 sea lions were examined, dying between 1 day and 10 months after admission to a marine mammal rehabilitation center. Persistent seizures with obtundation were the main clinical findings. Frequent gross findings in animals dying acutely consisted of piriform lobe malacia, myocardial pallor, bronchopneumonia, and complications related to pregnancy. Gross findings in animals dying months after intoxication included bilateral hippocampal atrophy. Histologic observations implicated limbic system seizure injury consistent with excitotoxin exposure. Peracutely, there was microvesicular hydropic degeneration within the neuropil of the hippocampus, amygdala, pyriform lobe, and other limbic structures. Acutely, there was ischemic neuronal necrosis, particularly apparent in the granular cells of the dentate gyrus and the pyramidal cells within the hippocampus cornu ammonis (CA) sectors CA4, CA3, and CA1. Dentate granular cell necrosis has not been reported in human or experimental animal DA toxicity and may be unique to sea lions. Chronically, there was gliosis, mild nonsuppurative inflammation, and loss of laminar organization in affected areas. PMID- 15753473 TI - Viral antigen distribution in the respiratory tract of cattle persistently infected with bovine viral diarrhea virus subtype 2a. AB - Tissues were obtained at necropsy from the nasal vestibule, turbinates, nasopharynx, trachea, tracheobronchial bifurcation, and lung from each of 10 clinically healthy calves persistently infected (PI) with bovine viral diarrhea virus (BVDV) serotype 2a. Tissues from the nasal vestibule were obtained by biopsy from five additional PI calves. Formalin-fixed tissues were processed for immunohistochemistry to localize the distribution of BVDV throughout the respiratory tract. Antigen distribution and intensity were subjectively evaluated. Throughout the respiratory tract, mononuclear leukocytes, vascular smooth muscle, and endoneural and perineural cells had BVDV immunoreactivity (BVDV-IR). Multifocally, squamous and ciliated columnar epithelium throughout the respiratory tract contained weak to moderate BVDV antigen. Viral antigen was not seen in goblet cells. BVDV-IR in mixed tubuloalveolar glands of the nasal cavity was weak to strong in serous secretory cells and ductular epithelium. Chondrocytes of the concha often contained BVDV antigen diffusely. Nasal mucus secreting and tracheobronchial glands multifocally contained weak viral signal. In all cases, alveolar macrophages had moderate to strong BVDV-IR, whereas BVDV IR in alveolar epithelial cells was weak to moderate. BVDV was present in interalveolar leukocytes and mesenchymal cells. Results indicate that serous secretions of the nasal cavity, productive viral infection of epithelium, and infected leukocytes in respiratory secretions are likely major sources of infectious BVDV from PI calves. The presence of BVDV antigen in respiratory epithelium is, at least, indirect support for the notion that this virus predisposes PI cattle to secondary microbial infections. PMID- 15753474 TI - A prospective analysis of immunohistochemically determined estrogen receptor alpha and progesterone receptor expression and host and tumor factors as predictors of disease-free period in mammary tumors of the dog. AB - The immunohistochemically determined estrogen receptor (ER) alpha (ERalpha) and progesterone receptor (PR) status, as well as recognized, well-accepted prognostic indicators and host factors were prospectively analyzed in 84 cases of primary canine mammary carcinoma for their effect on disease-free period (recurrence free, metastasis free, or combined) (DFP) after an observation period of 18 months. The presence of one or both receptors, as well as tumor size, lymph node status, histologic grading, intravascular growth, and necrosis, were of prognostic value for DFP. In multivariate analysis, only tumor size and histologic grading proved to be independent prognosticators. None of the host factors analyzed were of prognostic value for DFP. ERalpha, PR, or both were detected in 173 out of 228 tumors: 70 ERalpha and PR; 5 ERalpha only; 98 PR only. Statistically significant differences regarding the presence of one or both receptors were observed between benign and malignant tumors and between complex, mixed, and simple histologic subtypes of benign and malignant tumors. In the group of malignant tumors (n=155), the presence of one or both receptors was more frequent in tumors smaller than 3 cm, without lymph node metastasis, with tubulopapillary rather than solid patterns of growth among simple carcinomas, of histologic grades I and II, without both intravascular growth and necrosis, and with lymphocyte cell infiltrates. The most frequent groups of hormone receptors positive tumors were the ERalpha-positive and PR-positive group among benign and the ERalpha-negative and PR-positive group among malignant tumors. PMID- 15753475 TI - Heterotopic kidney tissue in the lung of a free-living common dolphin (Delphinus delphis). AB - A spontaneous case of renal heterotopia involving the lung parenchyma of a free living, adult, female common dolphin (Delphinus delphis), which was found stranded alive on the North Adriatic Sea coast of Italy, is reported in this study. The lesion, slightly visible from the macroscopic point of view, had the histologic appearance of a "foreign tissue island," which was poorly demarcated from the surrounding pulmonary tissue. Within such an island, several regularly shaped and apparently mature kidney glomeruli and tubules could be observed, with no evidence of secondary tissue reaction. To the best of our knowledge, this should be the first description of heterotopic kidney tissue occurrence in the lung of any domestic or wild animal species. PMID- 15753476 TI - A case of canine apocrine sweat gland adenoma, clear cell variant. AB - A cutaneous mass at the base of the retroauricular region of a 4-year-old, female Golden Retriever was examined pathologically. Histologically, the mass formed multiple nodules consisting of a proliferation of large clear cells with abundant cytoplasm. Mitotic figures among the neoplastic cells were very sparse. The large clear cells were intensely positive for cytokeratins (AE1/AE4, cytokeratin 8 and 18) and moderately positive for lysozyme and contained periodic acid-Schiff positive granules in the cytoplasm. In addition, small flat cells lined the islands of neoplastic large clear cells, and these were strongly positive for alpha-smooth muscle actin and vimentin, and some were positive for cytokeratin (AE1/AE4), suggesting they were myoepithelial cells. No local recurrence or metastasis has been recognized during the 18 months since surgical excision. On the basis of these findings, the present tumor was diagnosed as apocrine sweat gland adenoma, clear cell variant. There have been few previous reports of canine apocrine adenomas showing a clear cell morphology. PMID- 15753477 TI - West Nile virus encephalitis in a dog. AB - In November 2002, a 2-year-old, spayed Maltese Terrier in central Mississippi was presented for an acute illness characterized by uncontrolled hyperactivity that rapidly progressed to generalized tremors, ataxia, and intermittent hyperthermia. Postmortem examination after a 2-week course revealed mild, multifocal, nonsuppurative meningo encephalitis, with focal necrosis in the medulla. Reverse transcriptase-nested-polymerase chain reaction for West Nile virus (WNV) was positive on brain and negative on other tissues. Immunohistochemistry was negative on all tissues. The clinical, postmortem, and laboratory findings are consistent with acute encephalitis due to WNV infection. WNV infection should be considered in dogs showing signs of encephalitis when and where WNV and mosquito vectors occur. PMID- 15753478 TI - Ovarian epithelioid trophoblastic tumor in a cynomolgus monkey. AB - Epithelioid trophoblastic tumor (ETT) is an unusual type of trophoblastic tumor, with features resembling carcinoma. In this study, we describe a 4-year-old cynomolgus monkey (Macaca fascicularis) showing, at necropsy, a lobulated mass replacing the left ovary and several nodular lesions within the lungs. Histologically, the mass in the ovary and lung metastases were characterized by nests of epithelioid cells, with intermingled, occasional, multinucleate tumor cells consistent with syncytiotrophoblasts and moderate amount of eosinophilic, hyaline-like material. Immunohistochemically, the tumor cells were diffusely positive for cytokeratins (AE1/AE3) and inhibin-alpha, but only focal immunoreactivity was observed for human chorionic gonadotropin, whereas placental alkaline phosphatase was always negative. On the basis of morphology and immunohistochemical reactivity, tumor cells were identified as intermediate trophoblast. PMID- 15753479 TI - Ovarian choriocarcinoma in a rhesus monkey associated with elevated serum chorionic gonadotropin levels. AB - A clinically normal, 3-year-old female rhesus monkey (Macaca mulatta), which was part of a routine toxicology study, had a mass in the right ovary with metastases to the adjacent mesentery and lungs. The histologic features and immunohistochemistry results were consistent with the diagnosis of choriocarcinoma. Neoplastic cell types included cytotrophoblast (positive for cytokeratin), syncytiotrophoblast (positive for human chorionic gonadotropin), and extravillous trophoblast (positive for human placental lactogen). Because the neoplasm was present in the ovary, the uterus was normal, and the animal was not currently pregnant, this was considered a primary ovarian neoplasm of germ cell origin. The monkey had elevated serum levels of chorionic gonadotropin at the beginning of the study, indicating that, as in women, choriocarcinomas in monkeys can be associated with increased gonadotropin levels and that the tumor was preexisting at the start of the toxicology study. PMID- 15753480 TI - Hepatocellular carcinoma with metastasis to the spleen in a Holstein cow. AB - Hepatocellular carcinoma (HCC) with metastasis to the spleen in a Holstein cow was studied by histopathologic and immunohistochemical methods. The tumor was characterized by a pseudoglandular (acinar) pattern with an associated fibrous stroma. Individual cells often had a "hepatoid" appearance but were interspersed with scattered cells exhibiting a clear, periodic acid-Schiff (PAS)-positive cytoplasm and small eccentric nuclei. This pattern was present in nodules found in both liver and spleen. Moreover, hepatoid tumor cells were positive for alpha fetoprotein. Immunohistochemical studies suggest that myofibroblasts were responsible for the production of fibrous septa surrounding the pseudoglandular structures of bovine HCC. In summary, our histologic and immunohistochemical findings support a diagnosis of primary HCC with splenic metastasis. Furthermore, the associated stromal response appears to be of a myofibroblast origin. The primary etiology of bovine HCC and the significance of the intralesional, PAS positive clear cells remain undetermined. PMID- 15753481 TI - Adrenal myelolipoma in a dog. AB - Abdominal ultrasound examination in an 11-year-old, intact, female Labrador dog with hepatic disease revealed a nodular swelling of the left adrenal gland. Hyperadrenocorticism was suspected, but endocrine tests were negative. At the owner's request, an adrenalectomy was performed. Grossly, a nodular mass protruded from the external surface of the left adrenal gland and in cut section was hemorrhagic and effaced the cortical and medullary regions. Histologic examination revealed a cortical neoplasm with medullary involvement. The mass was composed of well-differentiated adipose cells, megakaryocytes, hematopoietic cells, and macrophages containing hemosiderin deposits. A diagnosis of cortical adrenal myelolipoma was made. PMID- 15753482 TI - Pharmacogenetics and ethnicity: more complexities of personalized prescribing. PMID- 15753483 TI - Critical care in AJRCCM 2004. PMID- 15753484 TI - Cystic fibrosis, pediatrics, control of breathing, pulmonary physiology and anatomy, and surfactant biology in AJRCCM in 2004. PMID- 15753485 TI - Tuberculosis, nontuberculous lung infection, pleural disorders, pulmonary function, respiratory muscles, occupational lung disease, pulmonary infections, and social issues in AJRCCM in 2004. PMID- 15753486 TI - Diagnosis and initial management of nonmalignant diseases related to asbestos. PMID- 15753487 TI - 2004 asbestos disease guidelines ignore mass screening abuse. PMID- 15753489 TI - Four years at the helm of the CJA: the past, the new Self-Assessment Program and the future. PMID- 15753490 TI - Measuring organ dysfunction in the intensive care unit: why and how? PMID- 15753491 TI - Predicting the difficult laryngoscopic intubation: are we on the right track? PMID- 15753492 TI - The skills exodus - being bled dry! PMID- 15753493 TI - The changing pattern of anesthesia, 1954-2004: a review based on the content of the Canadian Journal of Anesthesia in its first half-century. AB - PURPOSE: In order to review changes and progress in anesthesiology in the second half of the 20th century, and to recognize the first half-century of the Journal's existence. SOURCE: The content of the Journal from its inauguration in 1954 through 2004 was reviewed. RESULTS: Although the data base is that of the Canadian Journal, many of the contributions were from other countries, and for this reason the findings will have relevance both in Canada and elsewhere. The review suggests that anesthesiology evolved in two phases in this period: from 1954 to 1978 and from 1979 to 2004. The first was characterized by the introduction of new drugs and adaptation to new surgical techniques; the second, by a greater emphasis on clinical excellence, outcome, quality patient care both in the operating room and elsewhere in the hospital, and research. CONCLUSIONS: Although profound advances in knowledge, techniques, and relationships, have shaped the pattern and practice of anesthesiology in this half-century, the basic concerns of anesthesiologists relating to the practice of anesthesia and to their patients remained unchanged. At the same time, the many advances that have shaped anesthesiology in this half-century have extended the understanding of the phenomenon of anesthesia and enhanced the quality of patient care, which gives rise to the hope that anesthesiologists will continue to fully achieve these twin goals in the next half-century. PMID- 15753494 TI - Propofol is superior to thiopental for intubation without muscle relaxants. AB - PURPOSE: To compare intubating conditions and cardiovascular changes following induction of anesthesia and tracheal intubation in patients receiving either lidocaine-remifentanil-propofol or lidocaine-remifentanil-thiopental prior to induction. METHODS: In a randomized, double-blind study 76 healthy adult patients were assigned to one of two groups: lidocaine 1.5 mg.kg(-1), remifentanil 2 mug.kg(-1) and propofol 2 mg.kg(-1) (Group P) or lidocaine 1.5 mg.kg(-1), remifentanil 2 mug.kg(-1) and thiopental 5 mg.kg(-1) (Group T). Ninety seconds after the administration of the hypnotic agent, laryngoscopy and tracheal intubation were attempted. Intubating conditions were assessed as excellent, good or poor on the basis of ease of ventilation, jaw relaxation, position of the vocal cords, and patient's response to intubation and slow inflation of the tracheal cuff. The mean arterial pressure (MAP) and heart rate (HR) were measured 45 sec after hypnotic agent administration, immediately after tracheal intubation, two and five minutes after intubation. RESULTS: Excellent intubating conditions were obtained in 84% of Group P patients and 50% of Group T patients (P < 0.05). The percentage decrease from baseline MAP was significantly higher in Group P than in Group T postinduction (27.4% +/- 11.6 vs 21.8% +/- 10.0) and immediately postintubation (19.0% +/- 16.7 vs 11.2% +/- 14.9). The percentage change from baseline HR was significantly higher in Group P than in Group T postinduction (13.8% +/- 9.7 vs 0.5% +/- 12.4), immediately postintubation (8.7% +/- 13.7 vs 2.1% +/- 13.1), and two minutes postintubation (7.04% +/- 14.3 vs 3.5% +/- 14.3). CONCLUSION: Lidocaine-remifentanil-propofol is superior to lidocaine-remifentanil-thiopental for tracheal intubation without muscle relaxants. However, it induces more hypotension and bradycardia. PMID- 15753495 TI - Landiolol attenuates tachycardia in response to endotracheal intubation without affecting blood pressure. AB - PURPOSE: Beta-adrenergic receptor antagonists (beta-antagonists) have long been used to control perioperative tachyarrhythmias. The effects of a beta(1) antagonist, landiolol, on perioperative hemodynamics are unknown. We aimed to determine the appropriate dosage of landiolol for the treatment of hemodynamic changes in response to endotracheal intubation. METHODS: Sixty-four patients without heart disease or hypertension, were assigned to receive saline (group C) or landiolol (0.1 or 0.3 mg.kg(-1); groups L1 and L3). Anesthesia was induced with propofol (2 mg.kg(-1) iv) followed by saline or landiolol iv. After ventilation with facemask using 2% sevoflurane in 100% oxygen for 90 sec, endotracheal intubation was performed. After intubation, anesthesia was maintained using 1% sevoflurane in combination with 50% nitrous oxide. Values of heart rate and mean arterial blood pressure were recorded before induction to five minutes after intubation. RESULTS: In group C, heart rate and mean blood pressure increased simultaneously after tracheal intubation, compared with baseline values. Heart rate values were attenuated immediately before as well as after intubation in group L3, compared with groups C and L1. Heart rate did not increase after tracheal intubation in group L1, compared with baseline. In contrast, mean arterial blood pressure values did not differ among groups. CONCLUSIONS: The newly developed beta(1)-antagonist landiolol (0.1 and 0.3 mg.kg( 1)) may help prevent tachycardia without affecting blood pressure during the induction of anesthesia. PMID- 15753496 TI - Prophylactic methylene blue in a patient with congenital methemoglobinemia. AB - PURPOSE: To report the beneficial effect of prophylactic methylene blue administration before induction of anesthesia in a patient with congenital methemoglobinemia. CLINICAL FEATURES: A 26-yr-old male patient known to have congenital methemoglobinemia was scheduled for turbinectomy under general anesthesia. The patient was clinically cyanotic with a pulse oximetry of 91%. Arterial blood gas analysis showed a partial pressure of oxygen (PaO(2)) of 81.3 mmHg associated with a fractional oxyhemoglobin of 80.7%, and a methemoglobin fraction of 0.159. Preoperative iv administration of 1 mg.kg(-1) of methylene blue resulted, within five minutes, in a decrease of methemoglobin fraction down to 0.05 associated with an increase of the fractional oxyhemoglobin saturation up to 94.7%. After two hours, the methemoglobin fraction decreased to 0.01 and the fractional oxyhemoglobin concentration increased to 97.7%. Induction of anesthesia as well as intraoperative and postoperative course were uneventful without any episode of hypoxemia. Postoperatively, the methemoglobin fractions remained low for 24 hr, to be followed by a gradual increase up to 0.02 on the second day to reach 0.094 on the fifth day. CONCLUSION: The prophylactic preoperative methylene blue administration in a patient with congenital methemoglobinemia significantly decreased the methemoglobin level and increased the fractional oxygen saturation with a consequent increase of the safety margin against perioperative hypoxemia. PMID- 15753497 TI - Hypocapnia attenuates mesenteric ischemia-reperfusion injury in a rat model. AB - PURPOSE: Hypocapnia, a recognized complication of high frequency oscillation ventilation, has multiple adverse effects on lung and brain physiology in vivo, including potentiation of free radical injury. We hypothesized that hypocapnia would potentiate the effects of mesenteric ischemia-reperfusion on bowel, liver and lung injury. METHODS: Anesthetized male Sprague-Dawley rats were ventilated with high frequency oscillation and were randomized to one of four groups, exposed to either mesenteric ischemia-reperfusion or sham surgery, and to either hypocapnia or normocapnia. RESULTS: All animals survived the protocol. Ischemia reperfusion caused significant histologic bowel injury. Bowel 8-isoprostane generation was greater in ischemia-reperfusion vs sham, but was attenuated by hypocapnia. Laser-Doppler flow studies of bowel perfusion confirmed that hypocapnia attenuated reperfusion following ischemia. Plasma alanine transaminase, reflecting overall hepatocellular injury, was not increased by ischemia-reperfusion but was increased by hypocapnia; however, hepatic isoprostane generation was increased by ischemia-reperfusion, and not by hypocapnia. Oxygenation was comparable in all groups, and compliance was impaired by ischemia-reperfusion but not by hypocapnia. CONCLUSION: Hypocapnia, although directly injurious to the liver, attenuates ischemia-reperfusion induced lipid peroxidation in the bowel, possibly through attenuation of blood flow during reperfusion. PMID- 15753498 TI - [Neurostimulation does not increase the success rate of saphenous nerve blocks]. AB - PURPOSE: To evaluate neurostimulation of motor components of the vastus medialis muscle and the rectus femoris muscle, with a view to blocking the medial sensory fibres of the saphenous nerve. METHOD: First we dissected four femoral trigones, in order to select our puncture point. We were able to observe that, at the flexion crease of the thigh, the different fibers that make up the femoral nerve were clustered together and the saphenous nerve and the vastus medialis nerve had not yet separated from the femoral stem. Secondly, we conducted a prospective clinical study among 71 patients who had undergone surgery on the lower third of the leg with a sciatic block and a saphenous nerve block. The saphenous nerve block was performed using 10 mL of local anesthetic, by puncturing the flexion crease of the thigh in a bid to obtain one of two muscle responses: medial (contraction of the vastus medialis muscle) or anterior (contraction of the rectus femoris muscle and elevation of the patella). RESULTS: There was an overall success rate of 80% with the saphenous nerve block, with no statistical difference existing between the two response types. Average duration for the block to be completed was two minutes and it took an average of 15 min before the anesthesia took effect. No complications were encountered, apart from a puncture of the femoral artery, which was clinically inconsequential. CONCLUSION: Neurostimulation of the vastus medialis muscle has the same effect as neurostimulation of the rectus femoris muscle with respect to anesthesia of the saphenous nerve. Neurostimulation of the medial compartment of the femoral nerve saves local anesthetic, compared to a standard femoral block. PMID- 15753499 TI - Tibial nerve block: evaluation of a novel midleg approach in 241 patients. AB - PURPOSE: Several techniques of tibial nerve (TN) block have been described but require mobilization of the patient. We describe a new landmark, along the internal tibial shaft edge at the midleg level, that allows to block the TN and to insert a catheter with the patient lying supine. METHODS: 241 ASA physical status I to III awake, supine patients were studied prospectively. Cutaneous projections of the internal tibial condyle and the internal malleolus were marked and the needle was inserted 45 degrees cephalad in an antero-posterior plane, midway on the line between those two points, 1 cm posterior to the tibial shaft's internal edge. The catheter was introduced in the peri-nervous space using nerve stimulation (< 0.5 mA) on both the Tuohy needle and catheter. Ten millilitres of 2% lidocaine were injected through the catheter. Cutaneous and dermatomal sensory blockade were assessed using cold and pinprick tests while motor block was assessed using a modified Bromage scale. Satisfaction and analgesia scores were noted after surgery for 48 hr. Adverse events were recorded. RESULTS: The TN was always blocked, matching the distal L5 cutaneous nerve supply. Blood reflux was present in five patients (needle or catheter). No additional adverse events were noted. During the initial postoperative 48 hr, 0.2% ropivacaine was infused through the catheter (5 mL.hr(-1)) which always provided effective pain relief. CONCLUSION: The midleg technique of TN anesthesia and catheter insertion allows patients to remain in the supine position and results in a high rate of homogeneous anesthesia, a low incidence of side effects and effective continuous analgesia. PMID- 15753500 TI - Rapid injection of epidural mepivacaine speeds the onset of nerve blockade. AB - PURPOSE: When used intraoperatively, mepivacaine can produce a satisfactory sensory block. However, insufficient information is available concerning the factors that affect the speed of nerve blockade with epidural analgesia. The optimal rate of injection of mepivacaine has not been determined. We examined whether the speed of epidural infusion of mepivacaine affects the speed of nerve blockade. METHODS: Forty patients, physical status ASA I-II, scheduled for gynecological abdominal surgery, were enrolled in this double blind randomized trial. A catheter was inserted 4 cm in the epidural space in the midline at L1 L2. Three minutes after a test dose of 2 mL plain 1% mepivacaine over four seconds, 8 mL were injected epidurally at a rate of 1 mL.sec(-1) (fast group) or 0.05 mL.sec(-1) (slow group). Sensory and motor blockade, blood pressure, and heart rate were assessed at five, ten, and 15 min after the epidural injection. RESULTS: There was a significant difference in the spread of sensory blockade at five minutes after the epidural injection between the two groups, but not at ten and 15 min. Blood pressure decreased at five and ten minutes, recovered at 15 min in the fast group, and remained stable in the slow group. CONCLUSION: Rapid injection of mepivacaine in the epidural space produced a more rapid onset of epidural block than slow injection, but there was no difference in the final extent of the block. PMID- 15753501 TI - Canadian pediatric anesthesiologists prefer inhalational anesthesia to manage difficult airways. AB - PURPOSE: To survey Canadian pediatric anesthesiologists to assess practice patterns in managing pediatric patients with difficult airways. METHODS: Canadian pediatric anesthesiologists were invited to complete a web survey. Respondents selected their preferred anesthetic and airway management techniques in six clinical scenarios. The clinical scenarios involved airway management for cases where the difficulty was in visualizing the airway, sharing the airway and accessing a compromised airway. RESULTS: General inhalational anesthesia with spontaneous respiration was the preferred technique for managing difficult intubation especially in infants (90%) and younger children (97%), however, iv anesthesia was chosen for the management of the shared airway in the older child (51%) where there was little concern regarding difficulty of intubation. Most respondents would initially attempt direct laryngoscopy for the two scenarios of anticipated difficult airway (73% and 98%). The laryngeal mask airway is commonly used to guide fibreoptic endoscopy. The potential for complete airway obstruction would encourage respondents to employ a rigid bronchoscope as an alternate technique (17% and 44%). CONCLUSION: Inhalational anesthesia remains the preferred technique for management of the difficult pediatric airway amongst Canadian pediatric anesthesiologists. Intravenous techniques are relatively more commonly chosen in cases where there is a shared airway but little concern regarding difficulty of intubation. In cases of anticipated difficult intubation, direct laryngoscopy remains the technique of choice and fibreoptic laryngoscopy makes a good alternate technique. The use of the laryngeal mask airway was preferred to facilitate fibreoptic intubation. PMID- 15753502 TI - Modified Mallampati test, thyromental distance and inter-incisor gap are the best predictors of difficult laryngoscopy in West Africans. AB - PURPOSE: To determine the ability to predict difficult visualization of the larynx (DVL) from the following preoperative airway predictive indices, in isolation and combination: modified Mallampati test (MMT), thyromental distance (TMD), sternomental distance (SMD), horizontal length of the mandible (HLM) and inter-incisor gap (IIG). METHODS: Three hundred and eighty consecutive patients undergoing general anesthesia were evaluated using the MMT, TMD, SMD, HLM and IIG and the cut-off points for the airway predictors were Mallampati III and IV; < or = 6.5 cm, < or = 13.5 cm, < or = 9.0 cm and < or = 4.0 cm respectively. During direct laryngoscopy, the laryngeal view was graded using the Cormack and Lehane (CL) classification. CL grades III and IV were considered difficult visualization. Sensitivity, specificity and positive predictive value for each airway predictor in isolation and in combination were determined. Logistic regression analysis was used to determine independent predictors of DVL. RESULTS: Difficulty to visualize the larynx was found in 13 (3.4%) patients. The sensitivity, specificity and the positive predictive value for the five airway predictors were: MMT (61.5%; 98.4%; 57.1%), TMD (15.4%; 98.1%; 22.2%), SMD (0%; 100%; 0%), HLM (30.8%, 76.0%; 4.3%) and IIG (30.8%; 97.3%; 28.6%). The best combination of predictors was MMT/TMD/IIG with a sensitivity, specificity and positive predictive value of 84.6%; 94.6%; 35.5% respectively. Logistic regression analysis showed that weight, MMT, IIG and TMD were independent predictors of DVL. CONCLUSION: MMT, TMD and IIG appear to provide the optimal combination in prediction of DVL in a West African population. PMID- 15753503 TI - Parker Flex-Tip are not superior to polyvinylchloride tracheal tubes for awake fibreoptic intubations. AB - PURPOSE: Difficulty can be encountered during advancement of the tracheal tube (TT) over the bronchoscope after successful endotracheal bronchoscopy due to impingement on laryngeal structures. A new TT, the Parker Flex-Tip (PFT), has been shown to be superior to polyvinylchloride (PVC) TTs in anesthetized, paralyzed patients with normal airways. However, no study to date has shown the superiority of the new tapered tip design in patients with difficult airways during awake fibreoptic intubations (AFOI). The purpose of this study was to compare the PFT with PVC TTs for AFOI in patients with difficult airways or unstable c-spines. CLINICAL FEATURES: In this prospective observational study, 111 patients with predicted or documented difficult airways, or unstable c-spines were assessed for ease of TT advancement during AFOI. First attempt success rates were 91% for PFT TTs and 84% for PVC TTs (P = NS). Resistance to TT advancement was none to mild and similar in both groups. Advancement without the need to rotate the TT 180 degrees was also similar in both groups (57% vs 53%). CONCLUSION: For AFOI in patients with difficult airways, the PFT is not superior to conventional PVC TTs. PMID- 15753504 TI - Limited ability of SOFA and MOD scores to discriminate outcome: a prospective evaluation in 1,436 patients. AB - PURPOSE: The multiple organ dysfunction (MOD) score and sequential organ failure assessment (SOFA) score are measures of organ dysfunction and have been validated based on the association of these scores with mortality. We sought to compare the performance of the SOFA and MOD scores in a large cohort of consecutive multisystem intensive care unit (ICU) patients. METHODS: Prospective automated daily measurements of MOD and SOFA scores were performed in 1,436 patients admitted to a multisystem ICU in the Calgary Health Region over a one-year period. Logistic regression modeling techniques were used to describe the association of SOFA and MODS with mortality. Receiver operator characteristic (ROC) curves were used to assess the model's discriminatory ability. RESULTS: For ICU and hospital mortality, there was very little practical difference between the SOFA and MOD scores in their ability to discriminate outcome as determined by the area under the ROC. However, compared to previous literature, the discriminatory ability of both scores in this population was weak. As well, the calibration of the models was poor for both scores. The SOFA cardiovascular component score performed better than the MOD cardiovascular component score in the discrimination of both ICU and hospital mortality. CONCLUSIONS: SOFA and MOD scores had only a modest ability to discriminate between survivors and non survivors. These results question the appropriateness of using organ dysfunction scores as a 'surrogate' for mortality in clinical trials and suggest further work is necessary to better understand the temporal relationship and course of organ failure with mortality. PMID- 15753505 TI - Management following resuscitation from cardiac arrest: recommendations from the 2003 Rocky Mountain Critical Care Conference. AB - PURPOSE: To propose a strategy for the management of patients admitted to critical care units after resuscitation from cardiac arrest. SOURCE: Prior to the conference relevant studies were identified via literature searches and brief reviews circulated on the following topics: glucose and blood pressure management; therapeutic hypothermia; prearrest outcome prediction; post-arrest outcome prediction; and management of myocardial ischemia. Two days were devoted to assessing evidence and developing a management strategy at the conference. Consensus opinion of conference participants [intensive care unit (ICU) physicians] was used when high grade evidence was unavailable. Additional literature searches and data grading were performed post-conference. PRINCIPAL FINDINGS: High grade evidence was lacking in most areas. Specific goals of treatment were proposed for: general care; neurologic care; respiratory care; cardiac care; and gastrointestinal care. There was adequate evidence to recommend therapeutic hypothermia for comatose patients who had witnessed ventricular fibrillation or ventricular tachycardia arrests. Conference participants supported extending therapeutic hypothermia to other presenting rhythms in selected circumstances. Additional goals included mean arterial pressure 80 to 100 mmHg, glucose 5 to 8 mmol.L(-1) using insulin infusions, and PaO(2) > 100 mmHg for the first 24 hr. Absent withdrawal to pain 72 hr after resuscitation should prompt consideration of palliative care. The level of evidence for other recommendations was low. CONCLUSIONS: The proposed management strategy represents an approach to manage patients in the ICU following resuscitation from cardiac arrest. Most of the recommendations are based on low grade evidence. Additional research is needed to improve the evidence base. A standard post-arrest management strategy could help facilitate future research. PMID- 15753506 TI - Hyperemia and impaired cerebral autoregulation in a surgical patient with diabetic ketoacidosis. AB - PURPOSE: We describe cerebral hyperemia and impaired cerebral autoregulation documented with transcranial Doppler (TCD) ultrasonography in an adult patient with diabetic ketoacidosis (DKA) and sepsis presenting for surgery. CLINICAL FEATURES: Middle cerebral artery flow velocity was increased relative to PaCO(2) (Vmca 52 cm.sec(-1); PaCO(2) 22 mmHg) and the autoregulatory index (ARI) was 0 prior to surgery. Twenty hours after admission and treatment, cerebral hyperemia resolved (Vmca 52 cm.sec(-1) ; PaCO(2) 35 mmHg) and cerebral autoregulation returned to normal (ARI 0.91). CONCLUSION: To our knowledge, this is the first description of impaired cerebral autoregulation in adult DKA. Our observations suggest a relationship between cerebral hyperemia and impaired cerebral autoregulation in DKA. PMID- 15753507 TI - Continuous renal replacement therapy improves renal recovery from acute renal failure. AB - BACKGROUND: Acute renal failure (ARF) occurs in up to 10% of critically ill patients, with significant associated morbidity and mortality. The optimal mode of renal replacement therapy (RRT) remains controversial. This retrospective study compared continuous renal replacement therapy (CRRT) and intermittent hemodialysis (IHD) for RRT in terms of intensive care unit (ICU) and hospital mortality, and renal recovery. METHODS: We reviewed the records of all patients undergoing RRT for the treatment of ARF over a 12-month period. Patients were compared according to mode of RRT, demographics, physiologic characteristics, and outcomes of ICU and hospital mortality and renal recovery using the Chi square, Student's t test, and multiple logistic regression as appropriate. RESULTS: 116 patients with renal insufficiency underwent RRT during the study period. Of these, 93 had ARF. The severity of illness of CRRT patients was similar to that of IHD patients using APACHE II (25.1 vs 23.5, P = 0.37), but they required significantly more intensive nursing (therapeutic intervention scale 47.8 vs 37.6, P = 0.0001). Mortality was associated with lower pH at presentation (P = 0.003) and increasing age (P = 0.03). Renal recovery was significantly more frequent among patients initially treated with CRRT (21/24 vs 5/14, P = 0.0003). Further investigation to define optimal timing, dose, and duration of RRT may be beneficial. CONCLUSIONS: Although further study is needed, this study suggests that renal recovery may be better after CRRT than IHD for ARF. Mortality was not affected significantly by RRT mode. PMID- 15753508 TI - Antiemetic efficacy of propofol at small doses for reducing nausea and vomiting following thyroidectomy. PMID- 15753509 TI - An unusual occurrence in transfusion of autologous whole blood and Pentaspan. Beware! PMID- 15753510 TI - Severe hypotension following spinal anesthesia in a patient treated with risperidone. PMID- 15753511 TI - Intraoperative neuromonitoring in cardiac surgical patients with severe cerebrovascular disease. PMID- 15753512 TI - A 20-joule electrical cardioversion applied directly to the heart elevates troponin I by at least 1.5 ng.mL-1. PMID- 15753513 TI - Evaluating intubating conditions using the GCRP recommendations. PMID- 15753514 TI - Prime the ProSeal drain tube with lube from a tube! PMID- 15753515 TI - Use of the laryngeal mask airway in a patient with a difficult airway during supra-stoma granuloma removal. PMID- 15753517 TI - Mainstem bronchial diameter for the left-sided Broncho-Cath double-lumen tube: an in vitro study. PMID- 15753516 TI - Cobra PLA can overcome LMA failure in patients with face and neck contractures. PMID- 15753518 TI - Retromolar tracheal tube positioning for patients undergoing faciomaxillary surgery. PMID- 15753519 TI - Blood conservation and the management of perioperative blood transfusions in a patient undergoing major vascular surgery: a Self-Assessment Program. PMID- 15753520 TI - Transglutaminases: new target molecules for inflammatory bowel disease? PMID- 15753521 TI - The complicated path to true causes of disease: role of nuclear factor kappaB in inflammatory bowel disease. PMID- 15753522 TI - Where there's smoke there's not necessarily fire. PMID- 15753523 TI - Thalidomide and cancer cachexia: old problem, new hope? PMID- 15753525 TI - Clue to a more serious diagnosis. PMID- 15753524 TI - Weakly acidic reflux in patients with chronic unexplained cough during 24 hour pressure, pH, and impedance monitoring. AB - BACKGROUND AND AIMS: Acid gastro-oesophageal reflux is one of the most important causes of chronic cough. The response to acid suppression in these patients is not as good as in patients with heartburn but improvement with antireflux surgery has been reported, suggesting the involvement of a non-acidic gastric component in the refluxate. Less acidic reflux may produce symptoms such as regurgitation or chest pain. We investigated whether chronic cough might be associated with weakly acidic reflux. METHODS: We studied 28 patients with chronic cough using 24 hour ambulatory pressure-pH-impedance monitoring. Manometry was used for precise recognition of cough and impedance-pHmetry to detect acid (pH <4), weakly acidic (pH 7-4), and weakly alkaline (impedance drops, pH >/=7) reflux. A symptom association probability (SAP) analysis was performed for each type of reflux. RESULTS: Analysis was completed in 22 patients with 24 cough events (5 92)/patient. The majority of cough events (69.4%) were considered "independent" of reflux whereas 30.6% occurred within two minutes of a reflux episode. Half of these (49%) were "reflux cough" sequences, involving acid (65%), weakly acidic (29%), and weakly alkaline (6%) reflux. Ten patients (45%) had a positive SAP between reflux and cough: five with acid, two with acid and weakly acidic, and three only with weakly acidic reflux. CONCLUSIONS: Ambulatory pressure-pH impedance monitoring with SAP analysis allowed precise determination of the temporal association between cough and gastro-oesophageal reflux (acid, weakly acidic, and weakly alkaline) and identification of a subgroup of patients with chronic cough clearly associated with weakly acidic gastro-oesophageal reflux. PMID- 15753527 TI - An unusual cause of abdominal pain in a patient with hepatitis C cirrhosis. PMID- 15753526 TI - Influence of erythromycin on gastric emptying and meal related symptoms in functional dyspepsia with delayed gastric emptying. AB - BACKGROUND AND AIMS: Although delayed gastric emptying is considered a major pathophysiological mechanism in functional dyspepsia, the efficacy of prokinetic drugs has not been established. Recent studies using macrolide prokinetics were negative but receptor desensitisation may have played a role. The aim of the present study was to evaluate the influence on meal induced symptoms of acutely administered erythromycin in patients with gastroparesis. METHODS: In 20 patients with functional dyspepsia, gastric emptying was studied twice using the (14)C octanoic acid and (13)C glycin breath test to establish the reproducibility of the test. Breath samples were taken before the meal and at 15 minute intervals for a period of 240 minutes postprandially. At each breath sampling, the patient was asked to grade the intensity (0-3) of six dyspeptic symptoms. Twenty four patients (three men, mean age 43.5 (3) years) with dyspeptic symptoms and delayed gastric emptying were studied twice after pretreatment with saline or erythromycin intravenously. RESULTS: Meal related symptom severity scores were reproducible. Treatment with erythromycin significantly enhanced solid and liquid gastric emptying (t(1/2) 146 (27) v 72 (7) minutes, respectively (p<0.01), and 87 (6) v 63 (5) minutes (p<0.001)). Only the severity of bloating was significantly improved by erythromycin (23 (3.9) v 14.5 (2.7); p<0.01); all other symptoms and the cumulative meal related symptom score were not altered by erythromycin. CONCLUSIONS: In a setting where desensitisation played no role, erythromycin enhanced gastric emptying was not associated with a beneficial effect on meal related symptom severity. PMID- 15753528 TI - Chromoendoscopic surveillance in hereditary diffuse gastric cancer: an alternative to prophylactic gastrectomy? AB - BACKGROUND: Hereditary diffuse gastric cancer (HDGC) is defined by germline mutations in the E-cadherin gene, CDH-1. The first family in which CDH-1 mutations were identified was a large Maori kindred, where lifetime penetrance is 70%. Prophylactic gastrectomy is an unacceptable option for many mutation carriers. The results of annual chromoendoscopic surveillance using the methylene blue/congo red technique in 33 mutation carriers over a five year period are described. PATIENTS AND METHODS: Thirty three confirmed CDH-1 mutation carriers (18 males, 15 females), median age 32 years (range 14-69), were enrolled in 1999 2003. Medical records, endoscopy, and pathology were reviewed retrospectively. RESULTS: Over five years, 99 surveillance endoscopies were performed, of which 93 were chromo-dye enhanced. Sixty nine chromoendoscopies were normal. In 24 procedures, 1-6 pale areas/stomach (size 2-10 mm) were detected post chromo-dye application (totalling 56 pale lesions). One biopsy was taken from each pale lesion: 23 lesions (41%) showed signet ring cell carcinoma (10 patients), 10 lesions (18%) gastritis (four patients), and 23 (41%) normal mucosa (10 patients). No chromo-dyes were used in six procedures with macroscopic lesions (two HDGC, four ulceration). Total gastrectomies from patients with carcinoma were macroscopically normal but pathological mapping showed multiple microscopic foci of early signet ring cell carcinoma. Correlation of chromoendoscopic and gastrectomy findings showed that congo red/methylene blue detected carcinoma foci 4-10 mm in size but not foci <4 mm. CONCLUSIONS: The use of chromoendoscopy following normal white light gastroscopy facilitated detection of early gastric carcinoma foci not visible with white light gastroscopy. If these findings are validated in other HDGC kindred, chromogastroscopy represents an improved surveillance technique that can be safely considered alongside prophylactic gastrectomy. PMID- 15753529 TI - Giant gastric ulcer: an endoscopic roller coaster. PMID- 15753530 TI - Homeostatic regulation of zinc transporters in the human small intestine by dietary zinc supplementation. AB - BACKGROUND: The role of intestinal transporter regulation in optimising nutrient absorption has been studied extensively in rodent and cell line models but not in human subjects. AIMS: The aim of the present study was to investigate the response in vivo of zinc transporters in the human enterocyte to dietary zinc supplementation. SUBJECTS: Eighteen patients who had previously undergone ileostomy, all free of any symptoms of inflammatory bowel disease. METHODS: Subjects took a daily zinc supplement of 25 mg for 14 days in a double blind, placebo controlled, crossover trial. The effect of the supplement on expression in ileal biopsies of the zinc transporters SLC30A1, SLC30A4, SLC30A5, SLC39A1, SLC39A4, and metallothionein was measured by reverse transcription-polymerase chain reaction RT-PCR. Expression of SLC30A1, SLC30A5, and SLC39A4 was also examined by immunoblotting. RESULTS: The zinc supplement reduced SLC30A1 mRNA (1.4-fold) together with SLC30A1, SLC30A5, and SLC39A4 protein (1.8-fold, 3.7 fold, and to undetectable levels, respectively) in ileal mucosa and increased metallothionein mRNA (1.7-fold). The supplement had no effect on expression of SLC30A4 or SLC39A1 mRNA. Localisation of SLC30A5 at the apical human enterocyte/colonocyte membrane and also at the apical membrane of Caco-2 cells was demonstrated by immunohistochemistry. Commensurate with these observations in zinc supplemented human subjects, SLC30A1, SLC30A5, and SLC39A4 mRNA and protein were reduced in Caco-2 cells cultured at 200 muM compared with 100 muM zinc. CONCLUSIONS: These observations indicate that, in response to variations in dietary zinc intakes, regulated expression of plasma membrane zinc transporters in the human intestine contributes to maintenance of zinc status. PMID- 15753531 TI - An unusual abdominal mass. PMID- 15753532 TI - The RANKL/OPG system is activated in inflammatory bowel disease and relates to the state of bone loss. AB - BACKGROUND AND AIMS: A substantial proportion of patients with inflammatory bowel disease (IBD) develops osteopenia and osteoporosis in the course of disease. Recent data from a mouse model of colitis suggest that the receptor activator of nuclear factor kappa B (RANKL)/osteoprotegerin (OPG) system may be responsible for bone loss. METHODS: We investigated the activation state of the RANKL/OPG system and its association with bone loss in human IBD. Plasma levels of OPG and RANKL were correlated with bone mineral density and current IBD therapy. Colonic secretion of OPG and RANKL and cell types responsible for such secretion were determined. RESULTS: OPG plasma levels were elevated 2.4-fold in Crohn's disease (CD) and 1.9-fold in ulcerative colitis (UC) whereas soluble RANKL (sRANKL) levels were not significantly different in IBD patients compared with healthy controls. High levels of OPG were released from colonic explant cultures (CEC) derived from inflamed IBD specimens, and colonic macrophages and dendritic cells costained for OPG. sRANKL levels from CEC were low both in IBD patients and healthy controls. Interestingly, increased expression of RANKL was mainly confined to cells in the lamina muscularis. A significant negative correlation was found between OPG plasma levels and femoral neck/lumbar spine bone mineral density. CONCLUSIONS: We have demonstrated that IBD is associated with alterations in the RANKL/OPG system. Applying results from a murine model of colitis associated bone loss, the constellation of OPG and sRANKL regulation observed in our study raises the possibility that RANKL/OPG may contribute to the development of bone loss in IBD. PMID- 15753533 TI - Profile of soluble cytokine receptors in Crohn's disease. AB - INTRODUCTION: Soluble cytokine receptors (sCRs) modulate the in vivo activity of cytokines. Deficient sCR production could participate in the pathogenesis and course of Crohn's disease (CD). The aim of the study was to examine the profile of sCRs in CD patients and their modulation by infliximab and corticosteroids. METHODS: We prospectively examined active CD patients (aCD) treated with either infliximab (n = 21) or corticosteroids (n = 9), CD patients in clinical remission (rCD, n = 20), ulcerative colitis patients (UC, n = 24), and healthy subjects (HS, n = 15). Cultures of colonic biopsies were also examined from CD inflamed (n = 8), CD non-inflamed (n = 7), and healthy mucosa (n = 8). Levels of tumour necrosis factor alpha (TNF-alpha), soluble TNF receptor I (sTNFRI), soluble TNF receptor II (sTNFRII), interleukin 1beta (IL-1beta), soluble IL-1 receptor I (sIL 1RI), soluble IL-1 receptor II (sIL-1RII), IL-6, soluble IL-6 receptor (sIL-6R), and sgp130 were measured using ELISA. RESULTS: Higher levels of sTNFRI (p<0.05, p<0.01), sTNFRII (p<0.01, p<0.01), sIL-1RI (p<0.05, NS), IL-6 (p<0.01, p<0.01), and sIL-6R (p<0.05, NS) were observed in aCD compared with rCD and HS. Interestingly, sIL-1RII (p<0.05, p<0.01) and sgp130 (p<0.01, p<0.01) were profoundly decreased in aCD compared with rCD and HS, and were negatively correlated with CRP. Deficient production of sIL-1RII was specific to CD (not observed in ulcerative colitis), and was further confirmed at the mucosal level. Infliximab decreased sTNFRII at one and four weeks (p<0.05) and enhanced sIL-6R levels at one week (p<0.05). Corticosteroids increased sIL-1RII levels at one week (p<0.05). CONCLUSION: CD is associated with dysregulated production of sCRs. Deficiency in sIL-1RII and sgp130 may be essential to CD pathogenesis. Their replacement through the use of fusion proteins could represent future alternative therapeutic strategies for CD. PMID- 15753535 TI - Activation of nuclear factor kappaB in colonic mucosa from patients with collagenous and ulcerative colitis. AB - BACKGROUND AND AIMS: Expression of inducible nitric oxide synthase (iNOS) is greatly upregulated in the colonic mucosa of patients with collagenous and ulcerative colitis. As the transcription factor nuclear factor kappaB (NFkappaB) is a major inducer of iNOS gene expression, we compared activation and transcriptional activity of NFkappaB in colonic mucosal biopsies from these patients. PATIENTS: Eight patients with collagenous colitis, six with relapsing ulcerative colitis, and eight with uninflamed bowel were studied. METHODS: NFkappaB DNA binding activity was assessed by electrophoretic mobility shift assay and inhibitor of NFkappaB (IkappaB) kinase (IKK) activity by immunocomplex kinase assay. In vivo recruitment of NFkappaB to the iNOS promoter was determined by chromatin immunoprecipitation analysis and transcriptional activity by NFkappaB gene expression profiling arrays. Cells showing NFkappaB activation were identified by immunohistochemistry. RESULTS: In collagenous and ulcerative colitis, as opposed to uninflamed bowel, IKKbeta activity and strong NFkappaB DNA binding gave rise to activation of identical NFkappaB subunits and recruitment of transcriptionally active p65 to the iNOS promoter. In collagenous colitis, activated NFkappaB was observed only in epithelial cells while up to 10% of lamina propria macrophages showed activation in ulcerative colitis. CONCLUSIONS: In collagenous and ulcerative colitis, colonic mucosal NFkappaB is activated and recruited to the iNOS promoter in vivo via an IKKbeta mediated pathway. As collagenous colitis is not associated with tissue injury, these data challenge the prevailing view that activation of NFkappaB per se mediates tissue injury. Our results suggest that downstream inflammatory reactions leading to tissue damage originate in lamina propria immune cells, as increased NFkappaB activity in collagenous colitis was localised solely in epithelial cells, but present also in macrophages in ulcerative colitis. PMID- 15753534 TI - Differential expression of multiple transglutaminases in human colon: impaired keratinocyte transglutaminase expression in ulcerative colitis. AB - BACKGROUND AND AIMS: Ulcerative colitis (UC) is characterised by refractory inflammatory ulceration and damage to the colon. The mechanisms underlying impaired healing have yet to be defined. As transglutaminase expression resulting in matrix protein cross linking is associated with increased wound healing in a rat model of colitis, we hypothesised that different types of transglutaminase might also play a role in UC. PATIENTS AND METHODS: Endoscopic and histological indices were studied in 26 patients with UC (10 active and 16 inactive) and in 20 normal controls undergoing colonoscopy. Transglutaminase activity was evaluated in plasma (factor XIIIa) by a radioenzymatic method. Factor XIIIa, tissue and keratinocyte transglutaminase protein content, and mRNA expression in the colon were evaluated by western blot analysis and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), respectively. Colonic location of transglutaminases and their reaction products, the epsilon-(gamma glutamyl)lysine bonds, was evaluated by immunohistochemistry using specific monoclonal antibodies. RESULTS: Transglutaminase activity was significantly lower in the plasma of patients with active UC (4.2 (2.4) mU/ml; p<0.05 v controls) than in those with inactive UC and controls (10.6 (2.2) and 12.1 (1.7) mU/ml). As shown by western blot, protein levels of tissue transglutaminase and factor XIIIa were unchanged in active UC compared with inactive disease and controls, while the keratinocyte form was reduced in active UC. Tissue transglutaminase and factor XIIIa immunostaining was strongly present in damaged areas colocalising with isopeptide bonds. In contrast, the keratinocyte form was almost absent in active UC and localised in the upper part of the crypts in normal subjects. RT PCR showed upregulation of tissue transglutaminase mRNA in active UC (320% compared with controls) while keratinocyte transglutaminase gene expression was downregulated in active UC. CONCLUSIONS: The results of the present study support the concept that, in the damaged colon, transglutaminases are needed in response to chronic injury and underline the key role of these enzymes in mucosal healing. PMID- 15753536 TI - Cigarette smoking accelerates progression of alcoholic chronic pancreatitis. AB - BACKGROUND: Smoking is a recognised risk factor for pancreatic cancer and has been associated with chronic pancreatitis and also with type II diabetes. AIMS: The aim of this study was to investigate the effect of tobacco on the age of diagnosis of pancreatitis and progression of disease, as measured by the appearance of calcification and diabetes. PATIENTS: We used data from a retrospective cohort of 934 patients with chronic alcoholic pancreatitis where information on smoking was available, who were diagnosed and followed in clinical centres in five countries. METHODS: We compared age at diagnosis of pancreatitis in smokers versus non-smokers, and used the Cox proportional hazards model to evaluate the effects of tobacco on the development of calcification and diabetes, after adjustment for age, sex, centre, and alcohol consumption. RESULTS: The diagnosis of pancreatitis was made, on average, 4.7 years earlier in smokers than in non-smokers (p = 0.001). Tobacco smoking increased significantly the risk of pancreatic calcifications (hazard ratio (HR) 4.9 (95% confidence interval (CI) 2.3-10.5) for smokers v non-smokers) and to a lesser extent the risk of diabetes (HR 2.3 (95% CI 1.2-4.2)) during the course of pancreatitis. CONCLUSIONS: In this study, tobacco smoking was associated with earlier diagnosis of chronic alcoholic pancreatitis and with the appearance of calcifications and diabetes, independent of alcohol consumption. PMID- 15753537 TI - Effect of intravenous N-acetylcysteine infusion on haemostatic parameters in healthy subjects. AB - BACKGROUND AND AIMS: N-acetylcysteine is used to treat paracetamol overdose but depresses the activity of plasma coagulation factors II, VII, and X, which are often used to assess liver injury. The aim of this study was to investigate the effect of N-acetylcysteine on haemostasis in normal volunteers. METHODS: Haemostatic parameters in 10 healthy subjects were analysed before and following intravenous infusion of therapeutic doses of N-acetylcysteine, as well as in vitro. RESULTS: N-acetylcysteine induced significant decreases in plasma levels of vitamin K dependent haemostatic proteins in vivo, being maximal at one hour following the start of infusion, with maximal decreases from 1.00 to 0.73 (0.67 0.79) (mean (95% confidence interval)), 0.66 (0.58-0.73), 0.81 (0.73-0.90), 0.64 (0.57-0.70), 0.74 (0.65-0.82), and 0.61 (0.54-0.67) for factor II, VII, IX, and X activities, protein C activity, and free protein S reactivity, respectively. These data suggest that N-acetylcysteine induces protein modifications affecting activity. Five subjects developed an adverse reaction to infusion of N acetylcysteine and these were associated with a rapid increase in levels of factor VIII and its carrier protein von Willebrand factor (vWf) from 1.0 to 1.85 (1.08-2.62) and 1.77 (0.83-2.71), respectively, which suggests that the allergic reaction induced release of vWf from endothelial cells. N-acetylcysteine did not affect factor VIII or vWf in subjects without adverse reactions, and nor did it affect factor V or antithrombin in any of the subjects. CONCLUSION: Therapeutic doses of N-acetylcysteine cause abnormal haemostatic activity, and this should be taken into account when using haemostatic function tests as an indicator of hepatic injury. PMID- 15753538 TI - Increased anandamide induced relaxation in mesenteric arteries of cirrhotic rats: role of cannabinoid and vanilloid receptors. AB - BACKGROUND AND AIMS: Anandamide is an endocannabinoid that evokes hypotension by interaction with peripheral cannabinoid CB1 receptors and with the perivascular transient receptor potential vanilloid type 1 protein (TRPV1). As anandamide has been implicated in the vasodilated state in advanced cirrhosis, the study investigated whether the mesenteric bed from cirrhotic rats has an altered and selective vasodilator response to anandamide. METHODS: We assessed vascular sensitivity to anandamide, mRNA and protein expression of cannabinoid CB1 receptor and TRPV1 receptor, and the topographical distribution of cannabinoid CB1 receptors in resistance mesenteric arteries of cirrhotic and control rats. RESULTS: Mesenteric vessels of cirrhotic animals displayed greater sensitivity to anandamide than control vessels. This vasodilator response was reverted by CB1 or TRPV1 receptor blockade, but not after endothelium denudation or nitric oxide inhibition. Anandamide had no effect on distal femoral arteries. CB1 and TRPV1 receptor protein was higher in cirrhotic than in control vessels. Neither CB1 mRNA nor protein was detected in femoral arteries. Immunochemistry showed that CB1 receptors were mainly in the adventitia and in the endothelial monolayer, with higher expression observed in vessels of cirrhotic rats than in controls. CONCLUSIONS: These results indicate that anandamide is a selective splanchnic vasodilator in cirrhosis which predominantly acts via interaction with two different types of receptors, CB1 and TRPV1 receptors, which are mainly located in perivascular sensory nerve terminals of the mesenteric resistance arteries of these animals. PMID- 15753539 TI - Prevalence and clinical significance of isotype specific antinuclear antibodies in primary biliary cirrhosis. AB - BACKGROUND: Antinuclear antibodies (ANA) giving a rim-like/membranous (RL/M) or a multiple nuclear dot (MND) pattern are highly specific for primary biliary cirrhosis (PBC). Aim and SUBJECTS: To assess the prevalence of PBC specific ANAs, their Ig isotype, and their clinical significance in 90 PBC patients from Greece and Spain. Twenty eight patients with chronic hepatitis C, 23 patients with systemic lupus erythematosus, and 17 healthy subjects were studied as controls. METHODS: PBC specific ANA reactivity was tested by indirect immunofluorescence using HEp2 cells as substrate and individual Ig class (IgG, IgA, IgM) and IgG subclass (IgG1, IgG2, IgG3, IgG4) specific antisera as revealing reagents. RESULTS: Fourteen of 90 (15.6%) PBC patients had PBC specific ANA reactivity when an anti-IgG (total) antiserum was used as the revealing reagent while 58 (64.4%) were positive when specific antisera to each of the four IgG isotypes were used. The prevailing isotype was IgG3 for MND and IgG1 for RL/M. PBC patients with specific ANA, in particular of the IgG3 isotype, had significantly more severe biochemical and histological disease compared with those who were seronegative. None of the controls was positive. CONCLUSIONS: Disease specific ANA are present in the majority of patients with PBC when investigated at the level of immunoglobulin isotype. PBC specific ANA, in particular of the IgG3 isotype, are associated with a more severe disease course, possibly reflecting the peculiar ability of this isotype to engage mediators of damage. PMID- 15753540 TI - Diabetes increases the risk of hepatocellular carcinoma in the United States: a population based case control study. AB - BACKGROUND: Diabetes has been associated with an increased risk of hepatocellular carcinoma (HCC) in studies of referred patients. This is the first population based case control study in the USA to examine this association while adjusting for other major risk factors related to HCC. METHODS: We used the Surveillance Epidemiology and End-Results Program (SEER)-Medicare linked database to identify patients aged 65 years and older diagnosed with HCC and randomly selected non cancer controls between 1994 and 1999. Only cases and controls with continuous Medicare enrollment for three years prior to the index date were examined. Inpatient and outpatient claims files were searched for diagnostic codes indicative of diabetes, hepatitis C virus (HCV), hepatitis B virus (HBV), alcoholic liver disease, and haemochromatosis. HCC patients without these conditions were categorised as idiopathic. Unadjusted and adjusted odds ratios were calculated in logistic regression analyses. RESULTS: We identified 2061 HCC patients and 6183 non-cancer controls. Compared with non-cancer controls, patients with HCC were male (66% v 36%) and non-White (34% v 18%). The proportion of HCC patients with diabetes (43%) was significantly greater than non-cancer controls (19%). In multiple logistic regression analyses that adjusted for demographics features and other HCC risk factors (HCV, HBV, alcoholic liver disease, and haemochromatosis), diabetes was associated with a threefold increase in the risk of HCC. In a subset of patients without these major risk factors, the adjusted odds ratio for diabetes declined but remained significant (adjusted odds ratio 2.87 (95% confidence interval 2.49-3.30)). A significant positive interaction between HCV and diabetes was detected (p<0.0001). Similar findings persisted in analyses restricted to diabetes recorded between two and three years prior to HCC diagnosis. CONCLUSIONS: Diabetes is associated with a 2-3-fold increase in the risk of HCC, regardless of the presence of other major HCC risk factors. Findings from this population based study suggest that diabetes is an independent risk factor for HCC. PMID- 15753541 TI - Thalidomide in the treatment of cancer cachexia: a randomised placebo controlled trial. AB - BACKGROUND: Proinflammatory cytokines, especially tumour necrosis factor alpha (TNF-alpha), play a prominent role in the pathogenesis of cancer cachexia. Thalidomide, which is an inhibitor of TNF-alpha synthesis, may represent a novel and rational approach to the treatment of cancer cachexia. AIMS: To assess the safety and efficacy of thalidomide in attenuating weight loss in patients with cachexia secondary to advanced pancreatic cancer. METHODS: Fifty patients with advanced pancreatic cancer who had lost at least 10% of their body weight were randomised to receive thalidomide 200 mg daily or placebo for 24 weeks in a single centre, double blind, randomised controlled trial. The primary outcome was change in weight and nutritional status. RESULTS: Thirty three patients (16 control, 17 thalidomide) were evaluated at four weeks, and 20 patients (eight control, 12 thalidomide) at eight weeks. At four weeks, patients who received thalidomide had gained on average 0.37 kg in weight and 1.0 cm(3) in arm muscle mass (AMA) compared with a loss of 2.21 kg (absolute difference -2.59 kg (95% confidence interval (CI) -4.3 to -0.8); p = 0.005) and 4.46 cm(3) (absolute difference -5.6 cm(3) (95% CI -8.9 to -2.2); p = 0.002) in the placebo group. At eight weeks, patients in the thalidomide group had lost 0.06 kg in weight and 0.5 cm(3) in AMA compared with a loss of 3.62 kg (absolute difference -3.57 kg (95% CI -6.8 to -0.3); p = 0.034) and 8.4 cm(3) (absolute difference -7.9 cm(3) (95% CI -14.0 to -1.8); p = 0.014) in the placebo group. Improvement in physical functioning correlated positively with weight gain (r = 0.56, p = 0.001). CONCLUSION: Thalidomide was well tolerated and effective at attenuating loss of weight and lean body mass in patients with cachexia due to advanced pancreatic cancer. PMID- 15753542 TI - Relationship between symptoms and disordered continence mechanisms in women with idiopathic faecal incontinence. AB - BACKGROUND AND AIMS: Anal sphincter weakness and rectal sensory disturbances contribute to faecal incontinence (FI). Our aims were to investigate the relationship between symptoms, risk factors, and disordered anorectal and pelvic floor functions in FI. METHODS: In 52 women with "idiopathic" FI and 21 age matched asymptomatic women, we assessed symptoms by standardised questionnaire, anal pressures by manometry, anal sphincter appearance by endoanal ultrasound and magnetic resonance imaging (MRI), pelvic floor motion by dynamic MRI, and rectal compliance and sensation by a barostat. RESULTS: The prevalence of anal sphincter injury (by imaging), reduced anal resting pressure (35% of FI), and reduced squeeze pressures (73% of FI) was higher in FI compared with controls. Puborectalis atrophy (by MRI) was associated (p<0.05) with FI and with impaired anorectal motion during pelvic floor contraction. Volume and pressure thresholds for the desire to defecate were lower, indicating rectal hypersensitivity, in FI. The rectal volume at maximum tolerated pressure (that is, rectal capacity) was reduced in 25% of FI; this volume was associated with the symptom of urge FI (p<0.01) and rectal hypersensitivity (p = 0.02). A combination of predictors (age, body mass index, symptoms, obstetric history, and anal sphincter appearance) explained a substantial proportion of the interindividual variation in anal squeeze pressure (45%) and rectal capacity (35%). CONCLUSIONS: Idiopathic FI in women is a multifactorial disorder resulting from one or more of the following: a disordered pelvic barrier (anal sphincters and puborectalis), or rectal capacity or sensation. PMID- 15753545 TI - Small bowel malignancy at diagnosis of coeliac disease. PMID- 15753544 TI - Infection, coagulation, and variceal bleeding in cirrhosis. PMID- 15753546 TI - ITPA genotyping test does not improve detection of Crohn's disease patients at risk of azathioprine/6-mercaptopurine induced myelosuppression. PMID- 15753547 TI - IgG food antibodies should be studied in similarly treated groups. PMID- 15753548 TI - Terminal ileal biopsies should not be used to document extent of colonoscopic examination. PMID- 15753549 TI - Hypergastrinaemia in patients infected with Helicobacter pylori treated with proton pump inhibitors. PMID- 15753550 TI - Identification of ferroportin disease in the Indian subcontinent. PMID- 15753551 TI - Influence of dietary factors on the clinical course of inflammatory bowel disease. PMID- 15753552 TI - IgG antibodies to foods in IBS. PMID- 15753553 TI - Microdissection and microcloning of plant chromosomes. AB - Cytogenetic and molecular tools play an increasingly important role in plant genome research. A number of interesting applications that involve chromosome painting, the relationship between specific chromosomes and specific linkage groups, the relationships between physical and genetic distances on linkage maps, and the isolation of genes of interest, have been the subjects of recently published research. The aim of this paper is to review the different techniques available for chromosome microdissection and microcloning, and their use for the study of plant genomes. The quality of chromosomal DNA obtained is considered, and some recent results from our laboratory are presented. PMID- 15753554 TI - Organization and evolution of highly repeated satellite DNA sequences in plant chromosomes. AB - A major component of the plant nuclear genome is constituted by different classes of repetitive DNA sequences. The structural, functional and evolutionary aspects of the satellite repetitive DNA families, and their organization in the chromosomes is reviewed. The tandem satellite DNA sequences exhibit characteristic chromosomal locations, usually at subtelomeric and centromeric regions. The repetitive DNA family(ies) may be widely distributed in a taxonomic family or a genus, or may be specific for a species, genome or even a chromosome. They may acquire large-scale variations in their sequence and copy number over an evolutionary time-scale. These features have formed the basis of extensive utilization of repetitive sequences for taxonomic and phylogenetic studies. Hybrid polyploids have especially proven to be excellent models for studying the evolution of repetitive DNA sequences. Recent studies explicitly show that some repetitive DNA families localized at the telomeres and centromeres have acquired important structural and functional significance. The repetitive elements are under different evolutionary constraints as compared to the genes. Satellite DNA families are thought to arise de novo as a consequence of molecular mechanisms such as unequal crossing over, rolling circle amplification, replication slippage and mutation that constitute "molecular drive". PMID- 15753555 TI - Unraveling the genome structure of polyploids using FISH and GISH; examples of sugarcane and banana. AB - We review here the progress that has been achieved using molecular cytogenetics to analyze the genome structure of sugarcane (Saccharum spp) and banana (Musa spp), two crops that are polyploid, of interspecific origin and with chromosomes not distinguishable by their gross morphology. In Saccharum, molecular cytogenetics enabled us to determine the basic chromosome number of two species, Saccharum officinarum and S. spontaneum, involved in the origin of modern cultivars, to quantify the proportion of chromosomes of these species in the genome of modern cultivars, to assess the extent of interspecific chromosome recombination and to clarify the origin of the related species S. barberi. These techniques are also used to monitor introgression with related genera. In Musa, GISH enabled us to differentiate the four genomes involved in banana cultivars and allowed us to determine the genome constitution of several cultivars. FISH was used to analyze the distribution of repeated sequences along the genome. PMID- 15753556 TI - Diversity of a major repetitive DNA sequence in diploid and polyploid Triticeae. AB - About 90 members of a major tandemly repeated DNA sequence family originally described in rye as pSc119.2 have been isolated from 11 diploid and polyploid Triticeae species using primers from along the length of the sequence for PCR amplification. Alignment and similarity analysis showed that the 120-bp repeat unit family is diverse with single nucleotide changes and few insertions and deletions occurring throughout the sequence, with no characteristic genome or species-specific variants having developed during evolution of the extant genomes. Fluorescent in situ hybridization showed that each of the large blocks of the repeat at chromosomal sites harboured many variants of the 120-bp repeat. There were substantial copy number differences between genomes, with abundant sub terminal sites in rye, interstitial sites in the B genome of wheat, and relatively few sites in the A and D genome. We conclude that sequence homogenization events have not been operative in this repeat and that the common ancestor of the Triticeae tribe had multiple sequences of the 120-bp repeat with a range of variation not unlike that seen within and between species today. This diversity has been maintained when sites are moved within the genome and in all species since their divergence within the Triticeae. PMID- 15753557 TI - Variability of the chromosomal distribution of Ty3-gypsy retrotransposons in the populations of two wild Triticeae species. AB - Here, we report data on the population variability of Ty3-gypsy retrotransposons in genomes of Aegilops speltoides (2n = 2x = 14) and Hordeum spontaneum (2n = 2x = 14). Based on the sequence analysis or reverse transcriptase (RT) gene conserved domains, two groups of elements were recognized. Elements of Group I show relatedness to such a known element as RIRE2, and elements of Group II show relatedness to Fatima and Cereba. Cloned and sequenced fragments of Ty3-gypsy RT that show the closest relatedness to known elements (Fatima and RIRE2) were used as probes for fluorescent in situ hybridization (FISH). FISH experiments revealed mini-cluster organization of the Ty3-gypsy element chromosomal distribution in wild Triticeae species. Mini-clusters can be divided into three categories according to their intraspecific stability: (i) stable species-specific clusters that are mainly adjusted to the regions of rRNA genes; (ii) variable clusters that represent 68% of clusters in the genome of Ae. speltoides and 20% in the genome of H. spontaneum; and (iii) population-specific clusters that are mainly insertions into centromeric central domains of different chromosomes and the majority of these insertions were detected in populations with hot, dry environments. Significant interpopulation variability of Ty3-gypsy element chromosomal distribution in the Ae. speltoides genome contrasts with the uniform genome of H. spontaneum and may reflect differences in adaptive strategies between investigated species. PMID- 15753558 TI - Nuclear genome size and genomic distribution of ribosomal DNA in Musa and Ensete (Musaceae): taxonomic implications. AB - Nuclear DNA content and genomic distributions of 5S and 45S rDNA were examined in nineteen diploid accessions of the genus Musa representing its four sections Eumusa, Rhodochlamys, Callimusa and Australimusa, and in Ensete gilletii, which was the outgroup in this study. In the Eumusa (x = 11), 2C DNA content ranged from 1.130 to 1.377 pg, M. balbisiana having the lowest DNA content of all sections. M. beccarii (x = 9), a representative of Callimusa, had the highest 2C nuclear DNA content (1.561 pg). Species belonging to Rhodochlamys (x = 11) and Australimusa (x = 10) had 2C DNA contents ranging from 1.191 to 1.299 pg and from 1.435 to 1.547 pg, respectively. E. gilletii (x = 9) had 2C DNA content of 1.210 pg. The number of 5S rDNA loci in Musa varied from 4 to 8 per diploid cell. While different numbers of 5S rDNA loci were observed within Eumusa and Rhodochlamys, four 5S rDNA loci were observed in all accessions of Australimusa. M. beccarii (Callimusa) and E. gilletii contained 5S rRNA gene clusters on five and six chromosomes, respectively. The number of 45S rDNA loci was conserved within individual sections. Hierarchical cluster analysis of genome size, number of chromosomes and 45S rDNA sites suggested a close relationship between Rhodochlamys and Eumusa; Australimusa was clearly separated as were M. beccarii and E. gilletii. Within the Eumusa-Rhodochlamys group, M. balbisiana, M. schizocarpa and M. ornata formed distinct subgroups, clearly separated from the accessions of M. acuminata, M. mannii, M. laterita and M. velutina, which formed a tight subgroup. The results expand the knowledge of genome size and genomic distribution of ribosomal DNA in Musa and Ensete. They aid in clarification of the taxonomical classification of Musa and show a need to supplement the analyses on the DNA sequence level with cytogenetic studies. PMID- 15753559 TI - Long-range organization of plant satellite repeats investigated using strand specific FISH. AB - The technique of chromosomal orientation and direction fluorescence in situ hybridization (COD-FISH) was adapted for plant chromosomes in order to study long range organization of two families of satellite repeats, VicTR-B of Vicia sativa and PisTR-B of Pisum sativum. The technique allowed FISH to be performed on mitotic chromosomes in a strand-specific manner, resulting in visualization of the repeat orientation along the chromosomes and with respect to the direction of telomeric repeats. The VicTR-B probe applied to V. sativa chromosomes produced signals on a single chromatid at most regions containing corresponding sequences, thus confirming a presence of long arrays of head-to-tail arranged repeat monomers which is typical for satellite DNA. However, hybridization signals of different or equal intensities on both chromatids were also detected at some loci, suggesting a more complex arrangement of the repeats. Similar observations were made for PisTR-B repeats on P. sativum chromosomes, although the proportion of loci displaying signals on both chromatids was lower. In contrast to VicTR-B, orientation of the PisTR-B clusters with respect to telomeric sequences appeared to be conserved among subtelomeric regions of metacentric chromosomes and of the short arms of acrocentric chromosomes. PMID- 15753560 TI - Cytogenetic mapping in maize. AB - Cytogenetic maps depict the location and order of markers along chromosomes. Cytogenetic maps are important in genome research as they relate the genetic data and molecular sequences to the morphological features of chromosomes. In this paper, we discuss various methods used in cytogenetic mapping in maize, with special reference to fluorescence in situ hybridization (FISH) of single-copy sequences on meiotic pachytene chromosomes. PMID- 15753561 TI - 3D analysis of chromosome architecture: advantages and limitations with SEM. AB - Three-dimensional mitotic plant chromosome architecture can be investigated with the highest resolution with scanning electron microscopy compared to other microscopic techniques at present. Specific chromatin staining techniques making use of simultaneous detection of back-scattered electrons and secondary electrons have provided conclusive information on the distribution of DNA and protein in barley chromosomes through mitosis. Applied to investigate the structural effects of different preparative procedures, these techniques were the groundwork for the "dynamic matrix model" for chromosome condensation, which postulates an energy dependent process of looping and bunching of chromatin coupled with attachment to a dynamic matrix of associated protein fibers. Data from SEM analysis shows basic higher order chromatin structures: chromomeres and matrix fibers. Visualization of nanogold-labeled phosphorylated histone H3 (ser10) with high resolution on chromomeres shows that functional modifications of chromatin can be located on structural elements in a 3D context. PMID- 15753562 TI - High-resolution physical mapping of the secalin-1 locus of rye on extended DNA fibers. AB - High-resolution mapping of secalin-1 (Sec-1) locus has been performed by fluorescence in situ hybridization to extended DNA fibers of rye (Secale cereale, 2n = 14), employing DNA probes of lambda phage clones containing the omega secalin gene. The fluorescent signals to rye extended DNA fibers revealed continuous strings of 45 microm, corresponding to the size of 147 kb DNA. To determine the copy number of Sec-1 locus on DNA fibers, a 1.2-kb fragment including the entire coding region of the omega-secalin gene and a 1.0-kb fragment of the promoter region were amplified by PCR as probes for another fiber FISH. The physical position of these sequences was visualized as alternating fluorescent spots by multicolor in situ hybridization. Alternating signals of two DNA probes reflected the tandem repeated organization of the Sec-1 locus having 15 copies of the gene. The present findings based on fiber FISH analysis support the contention that the omega-secalin genes are arranged in a head-to-tail fashion separated by 8 kb of spacer sequences with a total length of 145 kb. PMID- 15753563 TI - Recent development of image analysis methods in plant chromosome research. AB - Image analysis methods have provided effective tools in chromosome research along with the development both in computer software and hardware. A chromosome image analyzing system, CHIAS, for plant chromosomes was developed in 1985 and was subsequently revised so that with CHIAS3 one can take advantage of Internet use for downloading the program. In this review, the recent developments of imaging methods in plant chromosome research for automating chromosome identification, constructing a map of a pachytene chromosome, and patterning of interphase nuclei are described. PMID- 15753564 TI - McClintock's controlling elements: the full story. AB - The chapter describes the early part of Barbara McClintock's work on DNA transposons in maize, in which she uncovered the Ac-Ds family of mobile "controlling elements". As a basis to understanding the work there is a preamble on the reproductive biology of maize, which explains the particular qualities of the maize plant as an experimental organism for genetics. An account is then given of the cytology of the system that was used to generate intact chromosomes having "sticky" (broken) ends, which is the starting point of the story. Cytogenetic aspects of the chromatid and chromosome breakage-fusion-bridge cycles, deriving from breakage, are then described; this leads on to a discussion about the way in which variegation phenotypes of the maize kernels can be "read" in terms of chromosome breakage. The "genetic earthquake" event of 1944, triggered by introducing broken chromosomes into a zygote from both parents, is then described; and the sequence of events leading to the discovery of Ds and Ac is traced. Finding mobility of Ds from one chromosomal location to another was pure serendipity, and the transposition showed itself while experiments were being undertaken to accurately map Ds. A similar chance observation revealed transposition of Ac as well, and then the relationship between the two elements was elucidated in terms of their autonomous and non-autonomous nature. The chapter concludes with a brief reference to the molecular cloning of Ac and Ds. PMID- 15753565 TI - Ribosomal DNA heterochromatin in plants. AB - The aim of this review is to integrate earlier results and recent findings to present the current state-of-the-art vision concerning the dynamic behavior of the ribosomal DNA (rDNA) fraction in plants. The global organization and behavioral features of rDNA make it a most useful system to analyse the relationship between chromatin topology and gene expression patterns. Correlations between several heterochromatin fractions and rDNA arrays demonstrate the heterochromatic nature of the rDNA and reveal the importance of the genomic environment and of developmental controls in modulating its dynamics. PMID- 15753566 TI - The positioning of rye homologous chromosomes added to wheat through the cell cycle in somatic cells untreated and treated with colchicine. AB - The arrangement of chromosome pairs 5RL and 7R added to the wild type and the ph1b mutant line of hexaploid wheat are analyzed in 2N somatic root tip cells during the cell cycle relative to the arrangement that chromosomes 5RL show in 4N tapetal cells produced after colchicine treatment. Both homologous chromosome pairs are identified at interphase and mitosis by fluorescence in situ hybridization. In nuclei at interphase, chromosomes appear as discrete domains that show the Rabl orientation. Homologous chromosomes are predominantly non associated and their positioning seems not to be influenced by the Ph1 gene that suppresses homoeologous meiotic pairing. This pattern of arrangement contrasts with the high level of somatic pairing that sister chromosomes show in the interphase that follows chromosome duplication induced by colchicine. Disruption of pairing observed in some 4N nuclei is produced at c-anaphase which suggests no topological redistribution of homologues during conformation of the new nucleus. Homologous chromosomes show no predominant arrangement in ellipsoidal metaphase plates, which contrasts with the preferential opposite location of homologues in human prometaphase rosettes. Differences between chromosomes in the variation of the length through the cell cycle and in the chromatin morphology when the Ph1 is absent suggest different patterns of chromatin condensation in both chromosomes. PMID- 15753567 TI - Movement ability of rye terminal neocentromeres. AB - Rye terminal neocentromeres were analyzed in various aspects. Plants with and without neocentromeres were crossed to determine the possible genetic control on their formation. The segregation obtained in our work is consistent with the hypothesis of two trans-acting genes determining neocentric activity in such a way that individuals with no neocentromeres at all would carry all non-activating alleles, whereas one activating allele might permit the activation of a few neocentromeres. Individuals with four activating alleles would show the maximum frequency of neocentromeres per cell. Anti-tubulin immunolabelling was used to visualize the interaction between the neocentromeres and the microtubules. In most cases an end-on interaction between neocentromeres and microtubules was observed, but a few neocentromeres were observed free of them. Spikes were irradiated at early meiosis to determine whether acentric fragments carrying subtelomeric heterochromatin were able to behave as neocentromeres. In no case were acentric fragments observed to form an extension polewards as they did in whole chromosomes. Broken chromosomes joined by a thin thread of chromatin to the centromeric region PMID- 15753568 TI - The simple ultrastructure of the maize kinetochore fits a two-domain model. AB - Light microscope observations suggest there are two discrete biochemical domains in the plant kinetochore, an inner domain containing structural proteins, and an outer domain containing proteins involved in motility. We analyzed the ultrastructure of maize meiotic kinetochores following high pressure freezing and freeze substitution, a method that provides excellent sample preservation. Data from meiosis II support previous descriptions of plant kinetochores as diffuse, nearly invisible domains, sometimes nesting in a cup of darkly staining chromatin. The ultrastructure is similar in meiosis I but there are two sister kinetochores that each protrude away from the chromosome and form their own distinct kinetochore fibers. Microtubules terminate within kinetochores where their ends are splayed in a cone-shaped configuration suggestive of microtubule disassembly. We could not detect any significant substructure within the kinetochore proper. We suggest that the diffuse structure classically defined as the kinetochore represents only the outer domain of a two-domain organelle. The inner domain, known to contain chromatin-binding proteins, probably extends into the electron-dense chromatin of the primary constriction. PMID- 15753569 TI - Molecular analysis of holocentric centromeres of Luzula species. AB - Luzula spp, like the rest of the members of the Juncaceae family, have holocentric chromosomes. Using the rice 155-bp centromeric tandem repeat sequence (RCS2) as a probe, we have isolated and characterized a 178-bp tandem sequence repeat (LCS1) from Luzula nivea. The LCS1 sequence is present in all Luzula species tested so far (except L. pilosa) and like other satellite repeats found in heterochromatin, the cytosine residues are methylated within the LCS1 repeats. Using fluorescent in situ hybridization (FISH) experiments we have shown that there are at least 5 large clusters of LCS1 sequences distributed at heterochromatin regions along each of the 12 chromosomes of L. nivea. We have shown that a centromeric antibody Skp1 co-localizes with these heterochromatin regions and with the LCS1 sequences. This suggests that the LCS1 sequences are part of regions which function as centromeres on these holocentric chromosomes. Furthermore, using the BrdU assay to identify replication sites, we have shown that these heterochromatin sites containing LCS1 associate when being replicated in root interphase nuclei. Our results also show premeiotic chromosome association during anther development as indicated by single-copy BAC in situ and the presence of fewer LCS1 containing heterochromatin sites in these cells. PMID- 15753570 TI - The controversial telomeres of lily plants. AB - The molecular structure of the exceptional telomeres of six plant species belonging to the order Asparagales and two species of the order Liliales was analyzed using Southern blot and fluorescence in situ hybridization. Three different situations were found, namely: i) In the two Liliales species, Tulipa australis (Liliaceae) and Merendera montana (Colchicaceae), the chromosome ends display hybridization signals with oligonucleotides resembling telomere repeats of both plants (TTTAGGG)n and vertebrates (TTAGGG)n. ii) Asparagales species such as Phormium tenax (Hemerocallidaceae), Muscari comosum (Hyacinthaceae), Narcissus jonquilla (Amaryllidaceae) and Allium sativum (Alliaceae) lack both the plant telomere repeats and the vertebrate telomere repeats. iii) Two other Asparagales species, Aloe vera (Asphodelaceae) and an Iris hybrid (Iridaceae), display positive hybridization with the vertebrate telomere repeats but not with the plant telomere repeats. Southern blot hybridization revealed concurring results. On this basis, the composition of the telomere structure in this plant group is discussed. PMID- 15753572 TI - Minichromosomes derived from the B chromosome of maize. AB - Fourteen minichromosomes derived from the B chromosome of maize are described. The centromeric region of the B chromosome contains a specific repetitive DNA element called the B repeat. This sequence was used to determine the transmission frequency of the different types of minichromosomes over several generations via Southern blot analysis at each generation. In general, the minichromosomes have transmission rates below the theoretical 50% frequency of a univalent chromosome. The gross structure of each minichromosome was determined using fluorescence in situ hybridization (FISH) on root tip chromosome spreads. The presence of the B centromeric repeat and of the adjacent heterochromatic knob sequences was determined for each minichromosome. In two cases, the amount of the centromeric knob repeat is increased relative to the progenitor chromosome. Other isolates have reduced or undetectable levels of the knob sequence. Potential uses of the minichromosomes are discussed. PMID- 15753573 TI - Differentiating plant cells switched to proliferation remodel the functional organization of nuclear domains. AB - The immature pollen grain, the microspore, under stress conditions can switch its developmental program towards proliferation and embryogenesis. The comparison between the gametophytic and sporophytic pathways followed by the microspore permitted us to analyse the nuclear changes in plant differentiating cells when switched to proliferation. The nucleus is highly dynamic, the architecture of its well organised functional domains--condensed chromatin, interchromatin region, nuclear bodies and nucleolus--changing in response to DNA replication, RNA transcription, processing and transport. In the present work, the rearrangements of the nuclear domains during the switch to proliferation have been determined by in situ molecular identification methods for the subcellular localization of chromatin at different functional states, rDNA, elements of the nuclear machinery (PCNA, splicing factors), signalling and stress proteins. The study of the changes in the nuclear domains was determined by a correlative approach at confocal and electron microscopy levels. The results showed that the switch of the developmental program and the activation of the proliferative activity affected the functional organization of the nuclear domains, which accordingly changed their architecture and functional state. A redistribution of components, among them various signalling molecules which targeted structures within the interchromatin region upon translocation from the cytoplasm, was also observed. PMID- 15753571 TI - Novel phosphorylation of histone H3 at threonine 11 that temporally correlates with condensation of mitotic and meiotic chromosomes in plant cells. AB - A novel mitosis-specific phosphorylation site in histone H3 at threonine 11 has been described for mammalian cells. This modification is restricted to the centromeric region while phosphorylation at the classical H3 sites, Ser10 and Ser28 occurs along the entire chromosomal arms. Using phosphorylation state specific antibodies we found that phosphorylation at threonine 11 occurs also in plant cells, during mitosis as well as meiosis. However, in contrast to animal cells, ph(Thr11)H3 was distributed along the entire length of condensed chromosomes, whereas H3 phosphorylated at Ser10 and Ser28 appeared to be restricted to centromeric/pericentromeric chromatin. Phosphorylation at Thr11 started in prophase and ended in telophase, it correlated with the condensation of mitotic and meiotic chromosomes and was independent of the distribution of late replicating heterochromatin and Giemsa-banding positive regions. Interestingly, treatment of cells with the phosphatase inhibitor cantharidin revealed a high level of Thr11 phosphorylation in interphase cells, in this case particularly in pericentromeric regions. These data show that histone modifications are highly dynamic. Moreover, animal and plant organisms may have evolved individual histone codes. PMID- 15753574 TI - Chromosome organization in wheat endosperm and embryo. AB - We have analysed the chromosome organization in endosperm and embryo of bread wheat (Triticum aestivum L.), in order to compare these tissues with developing anthers, in which the centromeres associate, and the developing root xylem vessel cells, in which the chromosomes endoreduplicate to become polytene and associate via their centromeres. Both endosperm and embryo showed a typical Rabl configuration and a degree of non-homologous centromere association and the endosperm also showed extensive telomere association. Wheat endosperm is initially triploid and during its development a percentage of the nuclei increase their DNA content to 6C and 12C. 6C nuclei showed twice as many centromeres as 3C nuclei and the centromere number increased further in 12C nuclei. The higher the C-content of a nucleus the more the telomeres associated in endosperm. The vast majority of 12C nuclei showed six rye chromosome arms, although a few showed three associated groups of rye chromosome arms. This means that during endosperm development wheat nuclei show both polyploidization and polytenization. PMID- 15753575 TI - A strategy to investigate the plant meiotic proteome. AB - The analysis of meiosis in higher plants has benefited considerably in recent years from the completion of the genome sequence of the model plant Arabidopsis thaliana and the development of cytological techniques for this species. A combination of forward and reverse genetics has provided important routes toward the identification of meiotic genes in Arabidopsis. Nevertheless identification of certain meiotic genes remains a challenge due to problems such as limited sequence conservation between species, existence of closely related gene families and in some cases functional redundancy between gene family members. Hence there is a requirement to develop new experimental approaches that can be used in conjunction with existing methods to enable a greater range of plant meiotic genes to be identified. As one potential route towards this goal we have initiated a proteomics-based approach. Unfortunately, the small size of Arabidopsis anthers makes an analysis in this species technically very difficult. Therefore we have initially focussed on Brassica oleracea which is closely related to Arabidopsis, but has the advantage of possessing significantly larger anthers. The basic strategy has been to use peptide mass-finger printing and matrix-assisted laser desorption ionization time of flight mass spectrometry to analyse proteins expressed in meiocytes during prophase I of meiosis. Initial experiments based on the analysis of proteins from staged anther tissue proved disappointing due to the low level of detection of proteins associated with meiosis. However, by extruding meiocytes in early prophase I from individual anthers prior to analysis a significant enrichment of meiotic proteins has been achieved. Analysis suggests that at least 18% of the proteins identified by this route have a putative meiotic function and that this figure could be as high as one-third of the total. Approaches to increase the enrichment of proteins involved in meiotic recombination and chromosome synapsis are also described. PMID- 15753576 TI - Plant chromosome homology: hypotheses relating rendezvous, recognition and reciprocal exchange. AB - Many higher eukaryotes have dispersed repetitive DNA and multiple instances of segmental duplications. As well, many plants and lower animals are polyploids. Thus restricting reciprocal genetic exchange to truly homologous chromosomes is likely a multi-step process. We propose the following sequence of events. First the ability to form a synaptonemal complex (SC) prematurely (i.e. before homology checking/recognition) is precluded by the organization of chromosomes during premeiotic S phase. Next rough alignment is accomplished regionally by having key allelic transcription units brought to the same transcription center. Once rough alignment is accomplished, close alignment can occur in conjunction with homology checking/recognition. Successful homology checking produces changes that now permit SC formation within the region of the check. Some organisms (with challenges to true homology such as dispersed repetitive DNA and segmental duplications) may require that, for a region to be competent to form an SC, successful homology checks must occur at both ends of the region. Successful early SC formation may provide an environment in which recombination intermediates can be earmarked for resolution into crossovers. Later in prophase I SC formation can occur nonhomologously, if two unsynapsed chromosomal axes meet. PMID- 15753578 TI - Understanding the cytological diploidization mechanism of polyploid wild wheats. AB - The allohexaploid Aegilops species (2n = 6x = 42), Ae. neglecta 6x (UUXtXtNN), Ae. juvenalis (DcDcXcXcUU), and Ae. vavilovii (DcDcXcXcSsSs) regularly form bivalents at metaphase I. However, in Ae. crassa 6x (DcDcXcXcDD) 0.27 quadrivalents per cell were observed probably as a consequence of the partial homology displayed by the D and Dc genomes. Likewise, the synthetic amphiploid Ae. ventricosa-Secale cereale (DDNNRR) is fertile and displays a diploid-like behavior at metaphase I, despite its recent origin. The pattern of synapsis at late zygotene and pachytene in the natural and artificial allohexaploids was analyzed by whole-mount surface-spreading of synaptonemal complexes under an electron microscope. It revealed that chromosomes were mostly associated as bivalents in all cases, the mean of multivalents per nucleus ranging from 0.17 (Ae. neglecta 6x) to 1.03 (Ae. crassa 6x) in the natural species and 1.05 in the Ae. ventricosa-S. cereale amphiploid. It can be concluded that the mechanism controlling bivalent formation in these species and also in the synthetic amphiploid acts mainly at zygotene by restricting synapsis to homologous chromosomes, but also acts at pachytene by preventing chiasma formation in the homoeologous associations. These observations are discussed in relation to the origin and evolution of the mechanism of diploidization in the allopolyploid species of the Poaceae family. PMID- 15753577 TI - Recombination nodules in plants. AB - The molecular events of recombination are thought to be catalyzed by proteins present in recombination nodules (RNs). Therefore, studying RN structure and function should give insights into the processes by which meiotic recombination is regulated in eukaryotes. Two types of RNs have been identified so far, early (ENs) and late (LNs). ENs appear at leptotene and persist into early pachytene while LNs appear in pachytene and remain into early diplotene. ENs and LNs can be distinguished not only on their time of appearance, but also by such characteristics as shape and size, relative numbers, and association with unsynapsed and/or synapsed chromosomal segments. The function(s) of ENs is not clear, but they may have a role in searching for DNA homology, synapsis, gene conversion and/or crossing over. LNs are well correlated with crossing over. Here, the patterns of ENs and LNs during prophase I in plants are reviewed. PMID- 15753579 TI - Synaptic behaviour of hexaploid wheat haploids with different effectiveness of the diploidizing mechanism. AB - Haploids of three cultivars of Triticum aestivum (Thatcher, Chris, and Chinese Spring) were obtained from crosses with Zea mays. The level of chromosome pairing at metaphase I and the synaptic behaviour at prophase I was studied. There were differences in the meiotic behaviour of the haploids from different cultivars. Thatcher and Chris haploids had significantly higher levels of pairing at metaphase I than Chinese Spring haploids. This metaphase I pairing was correlated with higher levels of synapsis achieved in the Thatcher and Chris prophase I nuclei than in the Chinese Spring nuclei. Variation in the effectiveness of the diploidizing mechanism among cultivars of wheat is proposed to have a genetic origin and the role of the Ph1 locus in the different haploids is discussed. PMID- 15753580 TI - Meiotic mutations in rye Secale cereale L. AB - Spontaneous meiotic mutations of winter rye Secale cereale L. (2n = 14) were revealed in inbred F2 progenies, which were obtained by self-pollination of F1 hybrids resulting from crosses of individual plants of cultivar Vyatka or weedy rye with plants of self-fertile inbred lines. The mutations cause partial or complete sterility, and are maintained in heterozygote condition. Six types of mutations were distinguished as the result of cytological analysis of meiosis and genetic analysis. (1) Plants with nonallelic asynaptic mutations sy1 and sy9 lacked bivalents in 96.8 and 67.0% metaphase I cells, respectively, formed only axial elements but not the mature synaptonemal complex (SC), and had defects in telomere clustering in early prophase I. (2) Weak asynaptic mutant sy3 showed incomplete synapsis at the start of SC degradation at diplotene and lower chiasma number; yet only 2% meiocytes lacked bivalents in MI. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19 caused nonhomologous synapsis; i.e., a varying number of univalents and occasional multivalents were observed in MI, which was preceded by switches of pairing partners and fold-back synapsis at mid-prophase I. (4) Mutation mei6 led to the formation of protrusions and minor branched structures of the SC lateral elements. (5) Allelic mutations mei8 and mei8-10 caused irregular chromatin condensation along the chromosome length in prophase I, which was accompanied by chromosome sticking and fragmentation in MI. (6) Allelic mutations mei5 and mei10 determined chromosome supercondensation, caused the disturbance of meiotic spindle assembly, arrested meiosis at various stages but did not affect formation of the pollen wall, thus arrested meiocytes got covered with the pollen wall. Analysis of double mutants revealed recessive epistatic interactions for some mutations; the epistatic group was sy9 > sy1 > sy3 > sy19. This reflects the sequence of meiotic events controlled by the corresponding genes. The expression of sy2 and sy19 proved to be modified by additional genes. Most meiotic mutations found in rye have analogs in other plants. PMID- 15753581 TI - Strategies for the study of meiosis in rye. AB - We describe how we are furthering our understanding of meiosis in rye (Secale cereale L.) using a combination of cytogenetic and molecular biological approaches. Fluorescent in situ hybridisation, electron microscopy of synaptonemal complexes, sequencing of meiosis-specific genes, and the immunolocalisation of recombinogenic proteins are being combined to build up phenotypic "identikits" of wild type, asynaptic mutants sy1 and sy9, and desynaptic mutant sy10. From this information, we review the status of our current understanding of the genetic control of meiosis in rye, and consider strategies for determining more precisely the interrelationships between meiosis specific genes and their products. PMID- 15753582 TI - Centromere-specific repetitive sequences from Torenia, a model plant for interspecific fertilization, and whole-mount FISH of its interspecific hybrid embryos. AB - Torenia fournieri is a good model plant to study fertilization in plants because it is easy to observe the fertilization process due to the protruding nature of the embryo sac. To study the location and movement of chromosomes and their centromeres in early embryogenesis of interspecific hybrid plants, we isolated two families of centromere-specific tandem repetitive sequences from T. fournieri and T. bailonii, and named them the "TCEN-family" and "BCEN-family", respectively. Both sequences consisted of a repeat unit of 52 bp located in the pericentric and centric heterochromatins. All signals of both sequences were prominent, but their intensity varied among the chromosomes. DNA-blot hybridization indicated the presence of similar sequences of TCEN-family in T. concolor, N. caerulea, and "Summer Wave", whereas the BCEN-family was found only in T. bailonii, thus indicating the wide or specific distribution of their repetitive families observed. We also applied whole-mount FISH to the interspecific hybrid embryos by using TCEN- and BCEN-family sequences as probes. Our results suggest that whole-mount FISH with the species-specific centromere sequences as probes is an ideal method to analyze the dynamics of chromosomes and centromeres in interspecific fertilization and early embryogenesis. PMID- 15753583 TI - Genome evolution of allopolyploids: a process of cytological and genetic diploidization. AB - Allopolyploidy is a prominent mode of speciation in higher plants. Due to the coexistence of closely related genomes, a successful allopolyploid must have the ability to invoke and maintain diploid-like behavior, both cytologically and genetically. Recent studies on natural and synthetic allopolyploids have raised many discrepancies. Most species have displayed non-Mendelian behavior in the allopolyploids, but others have not. Some species have demonstrated rapid genome changes following allopolyploid formation, while others have conserved progenitor genomes. Some have displayed directed, non-random genome changes, whereas others have shown random changes. Some of the genomic changes have appeared in the F1 hybrids, which have been attributed to the union of gametes from different progenitors, while other changes have occurred during or after genome doubling. Although these observations provide significant novel insights into the evolution of allopolyploids, the overall mechanisms of the event are still elusive. It appears that both genetic and epigenetic operations are involved in the diploidization process of allopolyploids. Overall, genetic and epigenetic variations are often associated with the activities of repetitive sequences and transposon elements. Specifically, genomic sequence elimination and chromosome rearrangement are probably the major forces guiding cytological diploidization. Gene non-functionalization, sub-functionalization, neo-functionalization, as well as other kinds of epigenetic modifications, are likely the leading factors promoting genetic diploidization. PMID- 15753584 TI - Allopolyploidy--a shaping force in the evolution of wheat genomes. AB - Recent studies have shown that allopolyploidy accelerates genome evolution in wheat in two ways: (1) allopolyploidization triggers rapid genome changes (revolutionary changes) through the instantaneous generation of a variety of cardinal genetic and epigenetic alterations, and (2) the allopolyploid condition facilitates sporadic genomic changes during the life of the species (evolutionary changes) that are not attainable at the diploid level. The revolutionary changes comprise (1) non-random elimination of coding and non-coding DNA sequences, (2) epigenetic changes such as DNA methylation of coding and non-coding DNA leading, among others, to gene silencing, (3) activation of genes and retroelements which in turn alters the expression of adjacent genes. These highly reproducible changes occur in the F1 hybrids or in the first generation(s) of the nascent allopolyploids and were similar to those that occurred twice in nature: first in the formation of allotetraploid wheat (approximately 0.5 million years ago) and second in the formation of hexaploid wheat (approximately 10,000 years ago). Elimination of non-coding sequences from one of the two homoeologous pairs in tetraploids and from two homoeologous pairs in hexaploids, augments the differentiation of homoeologous chromosomes at the polyploid level, thus providing the physical basis for the diploid-like meiotic behavior of allopolyploid wheat. Regulation of gene expression may lead to improved inter genomic interactions. Gene inactivation brings about rapid diploidization while activation of genes through demethylation or through transcriptional activation of retroelements altering the expression of adjacent genes, leads to novel expression patterns. The evolutionary changes comprise (1) horizontal inter genomic transfer of chromosome segments between the constituent genomes, (2) production of recombinant genomes through hybridization and introgression between different allopolyploid species or, more seldom, between allopolyploids and diploids, and (3) mutations. These phenomena, emphasizing the plasticity of the genome with regards to both structure and function, might improve the adaptability of the newly formed allopolyploids and facilitate their rapid and successful establishment in nature. PMID- 15753585 TI - The genome organization and diversification of maize and its allied species revisited: evidences from classical and FISH-GISH cytogenetic analysis. AB - The present review summarizes our classical and molecular cytogenetic investigations in the genus Zea. The results obtained from the meiotic behavior analysis of Zea species and hybrids, confirm the amphiploid nature of all species in the genus, with a basic number of x = 5 chromosomes. All species with 2n = 20 are diploidized allotetraploids, whereas Z. perennis (2n = 40) is an allooctoploid with four genomes somewhat divergent from one another. These analyses also revealed the existence of postzygotic reproductive isolation among Zea species. Our studies using genomic in situ hybridization (GISH) provide evidence about the evolutionary relationships among maize and its allied species, and reveal remarkable genomic divergences. Particularly, knob sequences were not completely shared between taxa previously considered to be closely related. Our data strongly suggest that the teosinte Z. mays parviglumis is not the only progenitor of cultivated maize. Introgression of Tripsacum into cultivated maize cannot be discarded. PMID- 15753586 TI - Architecture and evolution of dinoflagellate chromosomes: an enigmatic origin. AB - Dinoflagellates are a highly diversified group of unicellular protists that present fascinating nuclear features which have intrigued researchers for many years. As examples, a dense nuclear matrix accommodates permanently condensed chromosomes that are composed of fibers organized without histones and nucleosomes in stacked rows of parallel nested arches. The macromolecular chromosome structure corresponds to cholesteric liquid crystals with a constant left-handed twist. RNA acts to maintain the chromosome structure. Whole mounted chromosomes have a left-handed screw-like configuration with coils which progressively increase their pitch. This helical arrangement seems to be the result of a couple of narrow strands coiling together. Chromosomes do not show Q, G and C banding patterns. However, a roughly spherical differentiated upper end (primitive kinetochore?) and two differentiated coiling regions, the upper one composed of two to three coils where a couple of sister strands run together and parallel to each other, and the lower one where sister strands run out of phase by 180 degrees angular difference along the immediate next turns, can be distinguished. The chromosome segregation into two daughter chromatids begins at the telomere that attaches to the nuclear envelope, follows along the chromosome axis constituting first a Y-shaped and afterwards a V-shaped chromosome, which packs the newly synthesized DNA inside the "old" chromosome. Dividing chromosomes remain highly condensed, and the diameters of the new chromatids and the undivided chromosome are similar, but the number of arches is twice as large in G1 as in G2. The nuclear envelope remains through the cell cycle and shows spindle fibers, which penetrate intranuclear cytoplasmic channels during mitosis constituting an extra nuclear spindle. These and other cytogenetic features suggest that dinoflagellates are a group of enigmatic protists, unique and different from the usual eukaryotes. In contrast, DNA sequence studies propose that dinoflagellates are true eukaryotes, closely related to Apicomplexa, and ciliates (Alveolata), suggesting that the unusual features of chromosome and nuclear organization are not primitive but derived characters. Nevertheless, dinoflagellates have reached enigmatic specific nuclear and chromosome solutions, extremely far from those of other living beings. PMID- 15753587 TI - The relationships among lemons, limes and citron: a chromosomal comparison. AB - Lemons, limes and citron constitute a group of closely related Citrus species, whose species delimitations and taxonomic relationships are unclear. In order to identify karyotypic similarities and species relationships within this group, the CMA+/DAPI- banding pattern and the distribution of the 5S and 45S rDNA sites of 10 accessions of lime, lemon, and citron were investigated. The four cultivars of C. limon analyzed showed the same pattern of CMA+ bands and rDNA sites, suggesting that they originated from a single germplasm, later differentiated by distinct somatic mutations. The lemons C. jambhiri, C. limonia and C. volkameriana displayed karyotypes very similar to each other, but they differed from C. limon by the absence of a single chromosome with one band in each telomere. The limes, C. aurantifolia and C. limettioides, seemed less related to each other and exhibited different heteromorphic chromosome pairs. In C. aurantifolia, the presence of a chromosome type unknown in all other Citrus species cytologically known so far supports the assumption that this accession may be derived from a hybrid with a species from the subgenus Papeda or from another genus. Citrus medica was the only homozygous accession of this group and all of its chromosome types were clearly represented in limes and lemons, some of them forming heteromorphic pairs. The analysis of the distribution of rDNA sites allowed a further refinement of the comparison among accessions. The lemons and limes were heterozygous for all rDNA sites, whereas C. medica was entirely homozygous. These data support the hypothesis that C. medica is a true species while the other nine accessions are hybrids. PMID- 15753588 TI - Biogeographic distribution of polyploidy and B chromosomes in the apomictic Boechera holboellii complex. AB - The Boechera holboellii complex comprises B. holboellii and B. drummondii, both of which can reproduce through sex or apomixis. Sexuality is associated with diploid individuals, whereas apomictic individuals are diploid or triploid and may additionally have B chromosomes. Using flow cytometry and karyotype analysis, we have shown that B chromosomes (a) occur in both diploid and triploid apomictic B. holboellii, (b) may occur in triploid B. drummondii, and (c) are dispensable for the plant. Both diploid and triploid karyotypes are found in multiple chloroplast haplotypes of both species, suggesting that triploid forms have originated multiple times during the evolution of this complex. B chromosome carriers are found in geographically and genetically distinct populations, but it is unknown whether the extra chromosomes are shared by common descent (single origin) or have originated via introgressive hybridization and repeated transitions from diploidy to triploidy. Diploid plants containing the Bs reproduce apomictically, suggesting that the supernumerary elements are associated with apomixis. Finally, our analyses of pollen size and viability suggest that irregular chromosome segregation in some triploid lineages may lead to the generation of diploid individuals which carry the B chromosomes. PMID- 15753589 TI - Robertsonian translocations in wheat arise by centric misdivision of univalents at anaphase I and rejoining of broken centromeres during interkinesis of meiosis II. AB - The mechanism of origin of Robertsonian translocations was investigated in plants monosomic for chromosome 1A of wheat and 1H(t) of Elymus trachycaulus by GISH. Chromosomes 1A and 1H(t) stayed univalent in all metaphase I cells analyzed, suggesting that Robertsonian translocations do not originate from meiotic recombination in centromeric regions with shared DNA sequence homology. At ana /telophase I, the 1H(t) and 1A univalents underwent either chromosome or chromatid segregation and misdivided in 6-7% of the pollen mother cells. None of the ana-/telophases I analyzed had Robertsonian translocations, which were only observed in 2% of the "half tetrads" at ana-/telophase II. The frequency of Robertsonian translocations observed at ana-/telophase II corresponds well with the number of Robertsonian translocations (1-4%) detected in progenies derived from plants monosomic for group-1 chromosomes of wheat (1A, 1B, and 1D) and 1H(t) of E. trachycaulus. Our data suggest that Robertsonian translocations arise from centric misdivision of univalents at ana-/telophase I, followed by segregation of the derived telocentric chromosomes to the same nucleus, and fusion of the broken ends during the ensuing interkinesis. PMID- 15753590 TI - Molecular cytogenetics and tandem repeat sequence evolution in the allopolyploid Nicotiana rustica compared with diploid progenitors N. paniculata and N. undulata. AB - Nicotiana rustica (2n = 4x = 48) is a natural allotetraploid composed of P and U genomes which are closely related to genomes of diploid species N. paniculata and N. undulata. Genomic in situ hybridization (GISH) also confirms that the diploid parents, or close relatives, are the ancestors of N. rustica. In order to study genetic interactions between ancestral genomes in the allotetraploid, we isolated three families of repetitive sequences, two from N. paniculata (NPAMBE and NPAMBO) and one from N. undulata (NUNSSP). Southern blot hybridization revealed that the sequences are digested with a range of restriction enzymes into regular ladder patterns indicating a tandem arrangement of high copy repeats possessing monomeric units of about 180 bp. The three-tandem sequences belong to a larger Nicotiana tandem repeat family called here the HRS-60 family. Members of this family are found in all Nicotiana species studied. Fluorescence in situ hybridization (FISH) analysis localized the satellite repeats to subtelomeric regions of most chromosomes of N. paniculata and N. undulata. The pattern of sequence distribution on the P- and U-genomes of N. rustica was similar to the putative parents N. paniculata and N. undulata respectively. However, NPAMBO repeats appear to be reduced and rearranged in N. rustica that may suggest evolution within the P genome. GISH and FISH with the tandem repeat probes failed to reveal intergenomic translocations as might be predicted from the nucleocytoplasmic interaction hypothesis. PMID- 15753591 TI - Identification of individual chromosomes and parental genomes in Brassica juncea using GISH and FISH. AB - The three diploid (B. nigra, B. oleracea, B. campestris) and three allotetraploid (B. carinata, B. juncea, B. napus) species of Brassica, known as the "U-triangle" are one of the best model systems for the study of polyploidy. Numerous molecular investigations have provided a wealth of new insights into the polyploid origin and changes during the evolution of Brassica, but there are still many controversial aspects of their relationship and evolution. Interpretation of genome changes during evolution requires individual chromosome identification within the genome and clear distinction of genomes within the allotetraploid. The aim of this study was to identify individual chromosomes of B. juncea (genome AABB; 2n = 4x = 36) and to determine their genomic origin. Fluorescence in situ hybridization with 5S and 45S rDNA probes enabled discrimination of a substantial number of chromosomes, providing chromosomal landmarks for 20 out of 36 chromosomes of B. juncea. Additionally, along with double target genomic in situ hybridization, it allowed assignment of all chromosomes to either the A or B genomes. PMID- 15753592 TI - Wheat cytogenetics in the genomics era and its relevance to breeding. AB - Hexaploid wheat is a species that has been subjected to most extensive cytogenetic studies. This has contributed to understanding the mechanism of the evolution of polyploids involving diploidization through genetic restriction of chromosome pairing to only homologous chromosomes. The availability of a variety of aneuploids and the ph mutants (Ph1 and Ph2) in bread wheat also allowed chromosome manipulations leading to the development of alien addition/substitution lines and the introgression of alien chromosome segments into the wheat genome. More recently in the genomics era, molecular tools have been used extensively not only for the construction of molecular maps, but also for identification/isolation of genes/QTLs (including epistatic QTLs, eQTLs and PQLs) for several agronomic traits. It has also been possible to identify gene rich regions and recombination hot spots in the wheat genome, which are now being subjected to sequencing at the genome level, through development of BAC libraries. In the EST database also, among all plants wheat ESTs are the highest in number, and are only next to those for human, mouse, Ciona intestinalis (a chordate), rat and zebrafish genomes. These ESTs and sequences of several genomic regions have been subjected to a variety of applications including development of perfect markers and establishment of microcollinearity. The technique of in situ hybridization (including FISH, GISH and McFISH) and the development of deletion stocks also facilitated the preparation of physical maps. Molecular markers are also used for marker-assisted selection in wheat breeding programs in several countries. Construction of a wheat DNA chip, which will also become available soon, may further facilitate wheat genomics research. These enormous resources, knowledge base and the fast development of additional molecular tools and high throughput approaches for genotyping will prove extremely useful in future wheat research and will lead to development of improved wheat cultivars. PMID- 15753593 TI - Recent developments in durum wheat chromosome engineering. AB - Transfer of alien chromosome segments from various Triticeae species into cultivated wheats, commonly referred to as "chromosome engineering", is currently benefiting from the recent, impressive advancements in molecular genetics, cytogenetics and genomics, which are providing new insights into the genetic and physical organization of even complex plant genomes, such as those of the Triticeae. The powerful analytical tools presently available are making the assessment of desired genotypes in the course of chromosome engineering far more precise and effective than in the past, thus giving this transfer strategy renewed and increased potential for meaningful practical achievements. Examples are given here of the application of such tools to the engineering of the durum wheat genome with small alien segments containing genes with beneficial impact on disease resistance and quality traits. PMID- 15753594 TI - Production of alien chromosome additions and their utility in plant genetics. AB - Breeding programs aiming at transferring desirable genes from one species to another through interspecific hybridization and backcrossings often produce monosomic and disomic additions as intermediate crossing products. Such aneuploids contain alien chromosomes added to the complements of the recipient parent and can be used for further introgression programs, but lack of homoeologous recombination and inevitable segregation of the alien chromosome at meiosis make them often less ideal for producing stable introgression lines. Monosomic and disomic additions can have specific morphological characteristics, but more often they need additional confirmation of molecular marker analyses and assessment by fluorescence in situ hybridization with genomic and chromosome specific DNA as probes. Their specific genetic and cytogenetic properties make them powerful tools for fundamental research elucidating regulation of homoeologous recombination, distribution of chromosome-specific markers and repetitive DNA sequences, and regulation of heterologous gene expression. In this overview we present the major characteristics of such interspecific aneuploids highlighting their advantages and drawbacks for breeding and fundamental research. PMID- 15753595 TI - Recent progress in barley improvement using wild species of Hordeum. AB - In this review we describe recent progress in barley (Hordeum vulgare) improvement through hybridisation with its wild relatives. We have focused on one species in the secondary genepool of cultivated barley, namely H. bulbosum. This wild species has desirable traits, such as disease resistance, that are worthwhile transferring to its cultivated relative. Progress has been achieved through developing partially fertile interspecific hybrids that have been selfed or backcrossed to barley. We present the results of cytogenetic and molecular analyses that have enabled us to characterise and produce agronomically useful recombinant lines obtained from the hybrids. PMID- 15753596 TI - Detection of alien chromatin introgression from Thinopyrum into wheat using S genomic DNA as a probe--a landmark approach for Thinopyrum genome research. AB - The introduction of alien genetic variation from the genus Thinopyrum through chromosome engineering into wheat is a valuable and proven technique for wheat improvement. A number of economically important traits have been transferred into wheat as single genes, chromosome arms or entire chromosomes. Successful transfers can be greatly assisted by the precise identification of alien chromatin in the recipient progenies. Chromosome identification and characterization are useful for genetic manipulation and transfer in wheat breeding following chromosome engineering. Genomic in situ hybridization (GISH) using an S genomic DNA probe from the diploid species Pseudoroegneria has proven to be a powerful diagnostic cytogenetic tool for monitoring the transfer of many promising agronomic traits from Thinopyrum. This specific S genomic probe not only allows the direct determination of the chromosome composition in wheat Thinopyrum hybrids, but also can separate the Th. intermedium chromosomes into the J, J(S) and S genomes. The J(S) genome, which consists of a modified J genome chromosome distinguished by S genomic sequences of Pseudoroegneria near the centromere and telomere, carries many disease and mite resistance genes. Utilization of this S genomic probe leads to a better understanding of genomic affinities between Thinopyrum and wheat, and provides a molecular cytogenetic marker for monitoring the transfer of alien Thinopyrum agronomic traits into wheat recipient lines. PMID- 15753597 TI - Characterization of derivatives from wheat-Thinopyrum wide crosses. AB - Partial amphiploids are lines that contain 42 (38-42) wheat and 14 (14-18) alien chromosomes. They are derived by backcrossing wheat onto hybrids between wheat and either Thinopyrum intermedium (6x) or Th. ponticum (10x). GISH analysis has shown that, with possibly one exception, the alien genomes (chromosome sets) in partial amphiploids are found to be hybrids i.e. composed of chromosomes from more than one alien genome. The individual partial amphiploids are meiotically stable and nearly perfectly fertile, but hybrids between different lines were characterized by varying numbers of unpaired chromosomes and consequently variable degrees of sterility. Translocated chromosomes involving different Thinopyrum genomes or Thinopyrum and wheat genomes were found in partial amphiploids and consequently in the addition lines derived from them. Partial amphiploids have proven to be an excellent tertiary gene pool for wheat improvement, containing resistance to biotic stresses not present in wheat itself. Resistance to Barley Yellow Dwarf Virus (BYDV) and Wheat Streak Mosaic Virus (WSMV) have been found in partial amphiploids and addition lines derived from both Th. intermedium and Th. ponticum. Excellent resistance to Fusarium head blight has been found on a Th. intermedium chromosome that had substituted for chromosome 2D in wheat. Genes for resistance to leaf rust and stem rust have already been incorporated into wheat and tagged with molecular markers. PMID- 15753598 TI - Development and characterization of potato-Solanum brevidens chromosomal addition/substitution lines. AB - Solanum brevidens is a wild diploid potato species possessing high levels of resistances to several major potato diseases. We previously developed fertile somatic hybrids between S. brevidens and the cultivated potato (Solanum tuberosum) in order to introgress disease resistances from this wild species into potato. A series of backcross progenies was developed from a hexaploid somatic hybrid A206. Using a combination of S. brevidens-specific randomly amplified polymorphic DNA (RAPD) markers and a sequential genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH) technique, we identified all 12 S. brevidens chromosomes in the backcross progenies. Seven potato-S. brevidens monosomic chromosome addition lines (chromosomes 1, 3, 4, 5, 8, 9 and 10) and one monosomic substitution line (chromosome 6) were identified, and the remaining four S. brevidens chromosomes (2, 7, 11, and 12) were included in two other lines. These chromosomal addition/substitution stocks provide valuable tools for potato cytogenetic research, and can be used to introgress disease resistances from S. brevidens into potato. PMID- 15753599 TI - Limitations of in situ hybridization with total genomic DNA in routine screening for alien introgressions in wheat. AB - In situ hybridization with total genomic DNA (GISH) has become a powerful tool in characterization of alien introgressions in wheat. With recent simplification it can now be used in large scale screening for new chromosome constructs. Its level of resolution in routine applications was tested on sets of recombined wheat-rye chromosomes with genetically determined positions of the translocation breakpoints. The resolution level of GISH visualized by an enzymatic color reaction was much lower than that of GISH with fluorescent probes but both techniques failed to reveal the presence of some distally located breakpoints. The limits of resolution for the two methods were at least 9.8 and 3.5 cM of the relative genetic lengths of chromosome arms, respectively, in configurations with proximal rye and terminal wheat segments when rye DNA was used as a probe. When wheat DNA was used as a probe, a terminal wheat segment estimated to be ca. 1.6 cM in length could not be visualized. An example of induced recombination between a chromosome of Agropyron elongatum and wheat illustrates that these resolution limits of GISH may hamper isolation of critical translocation breakpoints in a chromosome engineering effort. PMID- 15753600 TI - Cytogenetics of Hordeum chilense: current status and considerations with reference to breeding. AB - Hordeum chilense Roem. et Schult. has a number of characteristics interesting for breeding: high crossability with other Triticeae, resistance to biotic and abiotic stresses and high variability for quality traits such as endosperm storage proteins or carotenoid content. xTritordeum, the amphiploids between H. chilense and different Triticum spp, are bridge species which facilitate the transfer of traits from H. chilense to wheat or triticale. The chromosome pairing between H. chilense and wheat chromosomes is very low (if existing) even in the absence of the action of the Ph1 gene. Nevertheless, translocation between H. chilense and wheat chromosomes has been observed frequently in genomic combinations where univalents of both species are present and therefore a method is available for using H. chilense in wheat or triticale breeding. Hybrids and amphiploids with other crop species of the Triticeae, such as rye or barley, have also been obtained, although to date the production of stable introgression stocks has not been completed. The technique of chromosome painting, using both high- and low-repeated DNA sequences in combination with genomic in situ hybridization have been used as effective methods for basic cytogenetic research in H. chilense, allowing analysis of genome evolution, and monitoring H. chilense chromosomes in interspecific hybridization breeding programs. PMID- 15753601 TI - Cytogenetics of Triticum x Dasypyrum hybrids and derived lines. AB - Genomic in situ hybridization was used to study Triticum x Dasypyrum wide hybrids and derived lines. A cytogenetic investigation was carried out in progenies of (i) amphiploids derived from T. turgidum var. durum (T. durum; 2n = 14; genomes AABB) x D. villosum (2n = 14; genome VV), (ii) three-parental hybrids (T. durum x D. villosum) x T. aestivum (2n = 42, genomes A'A'B'B'D'D'), and (iii) T. aestivum aneuploid lines carrying D. villosum chromosomes or chromatin. The amphiploids derived from T. durum x D. villosum showed a stable chromosomal constitution, made up of 14 V chromosomes, 14 chromosomes carrying the wheat A genome and 14 chromosomes carrying the B genome. High karyological instability was observed in the progenies of three-parental hybrids ([T. durum x D. villosum] x T. aestivum). Plants having the expected 14 A chromosomes, 14 B chromosomes, 7 D chromosomes, and 7 V chromosomes were rather rare (4.5%). Many progeny plants (45.5%) had the hexaploid wheat genome with 42 chromosomes and lacked any detectable D. villosum chromatin. Other plants (50%) had 14 A chromosomes and 14 B chromosomes, plus variable numbers of D and V chromosomes, the former being better retained than the latter in most cases. Some T. aestivum lines carrying D. villosum chromosomes or chromatin, as the result of addition, substitution, or recombination events or even a combination of these karyological events, were found to be stable. Other lines were unstable, and these lines carried 1V, 3V, or 5V chromosomes or their portions. Substitution or recombination events where 1V chromosomes were involved could concern the homeologous counterparts in both the A and B and D genomes of wheat. No line could be recovered where the shorter arm of 3V chromosomes was present. Changes in the morphology and banding pattern of V chromosomes were observed in hybrids that did not carry the entire D. villosum complement. By comparing the results of our cytogenetic analyses with certain phenotypic characteristics of the lines studied, genes for discrete traits could be assigned to specific V chromosomes or V chromosome arms. From the frequency of V chromosomes that were involved in chromatin exchanges with or substituted for one of their homeologous counterparts in the A, B, and D wheat genomes, it was inferred that D. villosum belongs to the same phyletic lineage as T. urartu (donor of the A genome of wheat) and Aegilops speltoides (B genome), and that Ae. squarrosa (D genome) diverged earlier from D. villosum. PMID- 15753602 TI - A decade of "chromosome painting" in Lolium and Festuca. AB - GISH has been a particularly useful technique for studying the Lolium-Festuca species complex of forage grasses. The reason for this utility is two-fold: (i) the complex is unique amongst crop plants in which fertile hybrids, and backcross progenies, can be produced which recombine genomes and promiscuously exchange their genes through homoeologous recombination; (ii) dispersed repetitive DNAs differ between species, and this allows tracking of the identity of chromosomes and chromosome segments. This tracking property has enabled several fruitful lines of research to produce a harvest of new information for both fundamental and practical purposes. We review this first decade of GISH (genomic in situ hybridization) in Lolium-Festuca, and discuss and summarize the achievements which have accrued. PMID- 15753603 TI - Central cell nuclear-cytoplasmic incongruity:a mechanism for segregation distortion in advanced backcross and selfed generations of (Allium cepa L. x Allium fistulosum L.) x A. cepa interspecific hybrid derivatives. AB - A model is presented as an explanation for an anomaly observed in germination and establishment and isozyme segregation patterns in Allium cepa x A. fistulosum F2BC3 populations generated in an introgression-breeding program. The F1BC3 parent of these populations was selected for its heterozygous PGI phenotype, Pgi 1(2/3); Pgi-1(2) was inherited from an A. cepa (Ac) seed parent and Pgi-1(3) from an A. fistulosum (Af) pollen parent. Germination and establishment was recorded for the F2BC3 progeny population. Segregation of Ac and Af Pgi-1 alleles was investigated in F2BC3 seeds and embryo and endosperm tissue was isolated and tested for isozyme expression. A pooled goodness-of-fit test of the segregation of Pgi-1 alleles in the populations to the expected Mendelian 1:2:1 ratio using the chi-square statistic gave a chi2 = 185.9, well beyond the accepted limits at 2 degrees of freedom. The 1:2:1 ratio expected for simple Mendelian inheritance was rejected, while a pooled chi-square goodness-of-fit test of the segregation of Pgi-1 alleles in the populations fit a 1:1 ratio with a chi2 = 0.203, based on the incongruity model. We present here the central cell nuclear-cytoplasmic incongruity hypothesis to explain the observed anomalies. PMID- 15753605 TI - Molecular analysis of pancreatic juice: a helpful tool to differentiate benign and malignant pancreatic tumors? AB - Chronic pancreatitis is an important predisposing condition leading to pancreatic carcinoma. As the differential diagnosis between these diseases may be difficult in 1 patient, the detection of specific tumor markers in pancreatic juice is an attractive diagnostic tool. Many studies have investigated tumor-mediated molecular alterations of the pancreatic juice, as k-ras mutations, telomerase reactivation, or promoter methylation of the tumor-suppressor genes p16INK4a and p14ARF. In this overview, we summarize these studies and conclude that molecular analysis of pancreatic juice is not useful for everyday care today. The high specificity of molecular alterations in pancreatic cancer in some pilot studies is waiting to be reproduced in large prospective trials, and has the potential to be a strong complementary marker of malignancy in patients with a pancreatic mass of uncertain dignity. PMID- 15753606 TI - The stress response of the exocrine pancreas. AB - Most attacks of acute pancreatitis display a self-limiting course. This suggests that pancreatic acinar cells may be able to protect themselves against cellular injury thus preventing further progression of the disease. In this review we describe several genes overexpressed in acute experimental pancreatitis which take part in the pancreatic stress response. We discuss the possible function of the pancreatitis-associated protein 1, the small nuclear protein p8, the glycoprotein clusterin, different heat shock proteins, the p53-dependent stress proteins TP53INP1alpha and TP53INP1beta, the vacuole membrane protein-1, as well as the interferon-inducible protein-15, the antiproliferative p53-dependent protein PC3/TIS21/BTG2, and the pancreatitis-induced protein-49. The implications of these proteins in pathophysiological processes like apoptosis regulation, regeneration, cell cycle and growth control, regulation of inflammation, and vacuole formation are discussed. Study of the function of stress proteins expressed in response to pancreatitis could widen our understanding of the pathophysiology of the disease and enable us to develop new rational therapeutic strategies. PMID- 15753607 TI - Laboratory markers of severe acute pancreatitis. AB - BACKGROUND: A large array of parameters has been proposed for the biochemical stratification of severity and prediction of complications in acute pancreatitis. However, the number of accurate and readily available variables for routine application is still limited. METHODS: The literature was reviewed for laboratory markers of acute pancreatitis with special regard to their clinical usefulness and test performance for stratifying severity and monitoring disease progression. RESULTS: Several parameters, such as trypsinogen and procarboxypeptidase B activation peptide, PMN-elastase, interleukin-6 (IL-6) and 8 (IL-8), serum amyloid A (SAA), and procalcitonin (PCT), can differentiate between mild and severe acute pancreatitis within 48 h of disease onset with favorable diagnostic accuracy. Because fully automated assays have become available, IL-6, IL-8, PCT, and SAA are the most interesting parameters in this respect. For monitoring disease progression beyond 48 h, acute-phase proteins, IL-6, IL-8, and PCT are valuable markers. PCT is the first biochemical variable for predicting severe pancreatic infections and overall prognosis throughout the course of acute pancreatitis with high sensitivity and specificity. CONCLUSIONS: Among all the biochemical variables available, C-reactive protein is still the standard for a fast, reliable, and cost-effective assessment of severity in acute pancreatitis. PCT substantially contributes to an improved stratification of patients at risk to develop major complications and deserves routine application. PMID- 15753608 TI - Tropical pancreatitis. AB - Tropical pancreatitis is a special type of chronic pancreatitis that is seen mainly in tropical countries. The prevalence of tropical pancreatitis is about 126/100,000 population in southern India. It occurs usually in young people, involves the main pancreatic duct and results in large ductal calculi. The etiology is not known, but genetic mutations such as the SPINK1 gene mutation and environmental factors are likely causes. Clinically, >90% of patients present with abdominal pain. About 25% of patients develop diabetes which generally requires insulin for its control but is ketosis-resistant. Painless diabetes is another clinical presentation in some patients. Most patients develop malnutrition during the course of the disease. Steatorrhea is less common. Patients with tropical pancreatitis may develop pancreatic cancer as a long-term complication. The diagnosis can be established by plain radiography of the abdomen, ultrasonography, computerized tomography scan of the abdomen or endoscopic retrograde cholangiopancreatography. Management is directed towards relief from pain and control of diabetes and steatorrhea. Pain relief can be obtained by analgesics and enzyme supplementation with preparations rich in proteases. Endotherapy coupled with stone fragmentation by extracorporeal shock wave lithotripsy is an effective therapy for those who fail to respond to medical therapy. Surgical decompression of the main pancreatic duct by lateral pancreato jejunostomy is reserved for patients with severe pain non-responsive to other forms of therapy. PMID- 15753609 TI - Pathogenesis of pain in chronic pancreatitis. AB - The pathophysiology of pain in chronic pancreatitis (CP) is incompletely understood. Several hypotheses have been advanced, including pancreatic and extrapancreatic causes. The existence of different hypotheses to explain the genesis of pain in CP also reflects the different therapeutic approaches to pain in these patients. Increased intraductal pressure as a result of single or multiple strictures and/or calculi is believed to be a common cause of pain in CP patients with a dilated main pancreatic duct. Other suggested causes include pancreatic fibrosis, interstitial hypertension and pancreatic ischemia. Additionally, extrapancreatic causes like duodenal and common bile duct stenosis with scarring due to pancreatic inflammation are suggested as factors causing pain in CP. The 'neurogenic inflammation' hypothesis is a fascinating theory which is supported by different studies. Immunohistological reports have shown that the amount of neurotransmitters, such as substance P and its receptor, calcitonin gene-related peptide and other neurotransmitters, are increased in afferent pancreatic nerves and a correlation between pain and immune cell infiltration of the nerves has been reported in CP. In this review we will discuss the different pain hypotheses and will present the perspective that neuroimmune interaction is an important factor for pain generation in CP. PMID- 15753610 TI - Mechanisms of pancreatic fibrosis. AB - Pancreatic fibrosis, a characteristic histopathological feature of chronic pancreatitis, is no longer considered an epiphenomenon of chronic injury, but an active process that may be reversible in the early stages. The identification and characterization of pancreatic stellate cells (PSCs) in recent years has had a significant impact on research into pancreatic fibrogenesis. Accumulating evidence from both in vivo studies (using human pancreatic sections and experimental models of pancreatic fibrosis) and in vitro studies (using cultured pancreatic stellate cells) indicates a key role for activated PSCs in the fibrotic process. These cells are now known to be activated by ethanol and its metabolites and by several factors that are upregulated during pancreatic injury including growth factors, cytokines and oxidant stress. Based on this knowledge, potential antifibrotic strategies such as antioxidants and cytokine inhibition have been assessed in experimental models. Studies are also underway to characterise the signaling pathways/molecules responsible for mediating PSC activation, in order to identify potential therapeutic targets for the inhibition of PSC activation, thereby preventing or reversing the development of pancreatic fibrosis. PMID- 15753611 TI - Endoscopic therapy of chronic pancreatitis. AB - We present an overview of endoscopic therapies for chronic pancreatitis (CP) and its associated conditions. It is evident that endoscopy can be a definite therapy for pancreatic pseudocysts, pancreatic ascites and pancreatic duct (PD) disruption. Endoscopic therapy has also been useful in the short-term and medium therapy of common bile duct strictures due to CP, the best results being obtained if there are no calcifications in the head of the pancreas. Although most experts agree that obstruction to the outflow of pancreatic juice and the resulting increased pressure within the main PD is one of the major factors contributing to pain and that endoscopic therapy has been proven effective to remove stones as well as to dilate PD strictures and place stents across the PD, there is no convincing evidence from randomized trials that the patient's dominant symptom of CP, i.e. pain, is resolved in an appropriate and long-term fashion. We believe that there are other factors which are important in the etiology of chronic pain such as pancreatic inflammation and peripancreatic fibrosis with resulting nerve entrapment around the gland. The reader is reminded that endoscopic therapy is associated with significant and important complications, therefore appropriate patient selection and patient information are of paramount importance. Nevertheless, it is important to consider that one advantage of endoscopic management of CP is that it is less invasive as compared with surgery, often effective for years, does not hinder further surgery, and can be repeated. Finally we want to emphasize that there are many valid surgical, radiological and endoscopic techniques to treat the complications of CP. Therefore, the approach to CP and its complications should be by a multidisciplinary team of gastroenterologists, surgeons, radiologists, endoscopists and pain specialists. PMID- 15753612 TI - Chronic parotitis: not another SPINKosis. AB - INTRODUCTION: Pancreatitis and parotitis share several etiological, pathohistological and functional similarities. It arose from recent pancreatitis research that some cases of chronic pancreatitis are associated with mutations of the serine protease inhibitor, Kazal type-1 (SPINK1). We tested the hypothesis that the pancreatitis-associated N34S mutation of SPINK1 is also a risk factor for chronic parotitis. METHODS: Reverse-transcriptase polymerase chain reaction was used to investigate SPINK1 transcription in the parotid gland. Forty-five blocks of formalin-fixed, paraffin wax-embedded tissues with chronic parotitis of unknown cause were analyzed for the SPINK1-N34S mutation. RESULTS: The SPINK1 gene is transcribed in the parotid gland. Two of the 45 patients (4.4%) with chronic parotitis carried the N34S mutation heterozygously. Of 82 healthy blood donors, 3 subjects (3.7%) were identified as carrying this mutation heterozygously (p = 0.83). CONCLUSION: The SPINK1-N34S mutation is not associated with chronic parotitis. PMID- 15753613 TI - Extended phenotype in the transthyretin Tyr77 familial amyloid polyneuropathy. AB - The transthyretin Tyr77 variant of familial amyloid polyneuropathy (FAP) has been identified in a few North American and European patients, but the full spectrum of its clinical manifestations is still not known. We report a 3-generation family of Jewish-Yemenite origin with Tyr77 FAP presenting with atypical features. The affected individuals had sensorimotor and autonomic neuropathy and cardiomyopathy accompanied by prominent dysphagia, hearing loss and asymptomatic carpal tunnel syndrome. Brain MRI in the proband showed multifocal white matter lesions. These features extend the reported Tyr77 phenotype and support the modifying effect of additional factors on the disease expression. PMID- 15753614 TI - Thymoma-associated neuromyotonia with antibodies against voltage-gated potassium channels presenting as chronic intestinal pseudo-obstruction. AB - Chronic intestinal pseudo-obstruction can occur as a paraneoplastic disorder, and several cases have been reported in association with thymoma or small-cell lung cancer. Autoantibodies against voltage-gated potassium channels (VGKCs) are found in acquired neuromyotonia (Isaac's syndrome), and have been reported in one case of slow transit constipation without apparent neurological disease. We describe a patient with VGKC antibodies, acquired neuromyotonia and thymoma, who first presented with a severe slow-transit constipation and in whom the gastrointestinal symptoms responded well to plasmapheresis. We suggest that VGKC antibodies might be helpful in patients with possible paraneoplastic chronic intestinal pseudo-obstruction, and a positive result should stimulate the search for a thymoma or other tumour and raise the possibility of immunotherapy. PMID- 15753615 TI - CD16+CD57- natural killer cells in multifocal motor neuropathy. AB - We analyzed the CD16+CD57- lymphocyte subset, which is considered to have strong natural killer (NK) cell activity, in peripheral blood from patients with chronic immune-mediated neuropathies and patients with other neurological diseases. We found that the ratio of CD16+CD57- NK cells to total lymphocytes was increased in 4 of 6 patients with multifocal motor neuropathy (MMN) with persistent conduction block. Since the CD16 molecule is an Fc receptor for immunoglobulin G (IgG), high dose intravenous immunoglobulin (IVIg) may interfere with CD16+CD57- NK cells via Fc receptor blockade. In addition, cyclophosphamide (Cy) is often used to suppress NK cells. Therefore, our findings may partly account for the effectiveness of IVIg or Cy, which is the current treatment of choice for MMN. PMID- 15753616 TI - Catechol-O-methyltransferase and monoamine oxidase B genes and susceptibility to sporadic Parkinson's disease in a Polish population. AB - Recent reports have proved that genetic factors play a role in the pathogenesis of sporadic Parkinson's disease (PD). It has been suggested that polymorphisms in monoamine oxidase B (MAOB) and catechol-O-methyltransferase (COMT) might increase the risk of PD. A total of 210 Polish patients with sporadic PD and 152 healthy controls were studied. The MAOB and COMT polymorphisms were identified using the polymerase chain reaction-restriction fragment length polymorphism method. The MAOB allele and genotype frequencies in PD patients did not differ significantly from the controls. A statistically lower frequency of the COMTLL genotype in patients with parkinsonism was found. The combined haplotype of the MAOB G (G/G) and COMTHL genotype showed a fourfold increase (p < 0.05) in the risk of PD in female patients in this Polish population. PMID- 15753617 TI - A multicenter, open-label study to evaluate satisfaction and menopausal quality of life in women using transdermal estradiol/norethindrone acetate therapy for the management of menopausal signs and symptoms. AB - AIM: This multicenter, open-label, single-arm study evaluated subject satisfaction and improvements in menopausal quality of life among menopausal women using the CombiPatch transdermal system consisting of 17beta-estradiol 0.05 mg plus norethindrone acetate 0.14 mg in a matrix patch formulation. METHODS: The 193 postmenopausal women between the ages of 45 and 65 years who comprised the modified intent-to-treat population (at least one patch and one efficacy assessment) were required to have reported at least five daily moderate-to-severe hot flashes and episodes of nocturnal sweating upon study entry for at least 1 month and applied one patch twice a week for 12 weeks. At weeks 0, 6, and 12, the women completed the Menopause-Specific Quality of Life (MENQOL) Questionnaire and, as a secondary study outcome, reported the scale of their application site discomfort. During weeks 1-12, they also kept diary records of number and severity of hot flashes and four other menopausal symptoms. Skin tolerance and adherence of the transdermal system were evaluated at weeks 6 and 12 by qualified evaluators. At week 6 and the end of the study, both subjects and physicians rated their satisfaction with the system. RESULTS: Among women in the modified intent-to-treat population, transdermal 17beta-estradiol plus norethindrone acetate significantly reduced the mean daily number of moderate-to-severe hot flashes experienced by women from 4.1 at week 1 to 0.6 at week 12 (p < 0.0001). The mean ratings of headache severity, insomnia, and vaginal irritation/dryness also improved significantly by week 6 and were maintained at week 12. At week 12, 92.4% of the subjects and 97.3% of the physicians reported that they were 'satisfied' or 'very satisfied' with the transdermal hormone delivery system. CONCLUSIONS: The results of this study compare favorably with previous placebo controlled studies of transdermal hormone therapy in managing menopausal signs and symptoms. Furthermore, quality of life was significantly improved by the transdermal hormone therapy system, and both subjects and physicians reported high levels of satisfaction. PMID- 15753618 TI - Obesity and Clomiphene Challenge Test as predictors of outcome of in vitro fertilization and intracytoplasmic sperm injection. AB - One hundred and sixty-two consecutive patients undergoing in vitro fertilization (IVF) or IVF/intracytoplasmic sperm injection (ICSI) were studied to determine the effect of obesity on the outcome of this treatment and to evaluate the prognostic value of the Clomiphene Challenge Test (CCT) in controlled ovarian hyperstimulation. In this prospective clinical study, we assessed the mean number of stimulation days, the mean gonadotropin level/day, cancellation rate, the mean thickness of the endometrium, the mean number of oocytes retrieved, the fertilization rate, the clinical pregnancy rate/embryo transfer and the abortion rate. Obese women had a doubled risk of cancellation due to poor response, although this was not statistically significant. Furthermore, they showed up to 45% lower fertilization rates compared to women of normal weight. The CCT was a good predictor of IVF and IVF/ICSI outcome. Women with an abnormal CCT needed more days of stimulation and higher doses of gonadotropins to reach an adequate stimulation, but still overall results were less than in women with a normal CCT. We conclude that obesity negatively affects IVF and IVF/ICSI outcome, and that CCT is a useful prognosticator of response to ovarian stimulation. Obese patients show a tendency to experience more cancellation due to poor response and lower fertilization rates. Obese women should be counseled on their possible poor performance in IVF and IVF/ICSI programs. PMID- 15753619 TI - Steroid and vasoactive treatment for acute deafness after attempted hearing preservation acoustic neuroma surgery. AB - OBJECTIVE: To investigate whether intravenous steroid and vasoactive therapy in the acute postoperative period improves hearing outcome in patients who develop acute deafness after attempted hearing preservation surgery for acoustic neuroma (AN) through a retrosigmoid or a middle cranial fossa approach. STUDY DESIGN AND SETTING: Retrospective controlled study in a tertiary care center. Thirty-six patients who had developed acute deafness after hearing preservation surgery for treatment of an AN were reviewed. Preoperative AAOHNS hearing class was A in 2, B in 2 and D in 32 patients. Twenty-seven patients were treated with prednisolone, hydroxyethyl starch 3% and pentoxifylline intravenously for a period of at least 5 days. Nine patients (controls) did not receive any specific steroid or vasoactive therapy. RESULTS: All patients in both groups remained deaf. CONCLUSIONS: Intravenous therapy with prednisolone, hydroxyethyl starch 3% and pentoxifylline in the acute postoperative period does not improve hearing in patients who develop acute deafness after attempted hearing preservation surgery for AN. PMID- 15753620 TI - A new ossicle homograft inactivation/preservation procedure: clinical results. AB - A new NaOH-autoclaving inactivation/preservation procedure (IPP) for ossicle homografts, complying with the actual infectious disease guidelines, has been developed and used in our institution for 5 years. This study compares the clinical and audiological results of middle ear reconstruction using the new NaOH autoclaving inactivated ossicle homografts (22 patients) and the previously used cialit-formaldehyde inactivation procedure (28 patients). During the follow-up period, no homograft extrusion, resorption or disease transmission was observed either for the NaOH-autoclaving or for the cialit-formaldehyde protocol. A postoperative air-bone gap of less than 20 dB in 44% and a postoperative hearing improvement of 10-50 dB in 70% of patients complies with the published success rates of homograft ossiculoplasty in the literature. The analysis and comparison of both tested IPP-patient groups showed no statistically significant differences in the clinical and the audiological results. The NaOH-autoclaving inactivation/preservation protocol should increase ossicle homograft safety even with respect to prion exposure. The good anatomic and audiological long-term results of the new IPP protocol confirm homograft ossicles as a valid and inexpensive approach for middle ear reconstruction. PMID- 15753621 TI - Effectiveness of the jaw-thrust maneuver in opening the airway: a flexible fiberoptic endoscopic study. AB - OBJECTIVE: A prospective study was carried out to find the exact site of obstruction in sleep model and to quantitatively evaluate the effect of Jaw thrust maneuver (JTM) in opening the obstructed airway using flexible fiberoptic endoscope. METHODS: Twenty-eight ASA physical status I or II patients with snoring symptom undergoing elective surgery were included. The patients were held in supine position without hyperextension of the neck. Having induced anesthesia, the base of the tongue and laryngeal inlet and/or epiglottis were visualized using endoscope. The patients' epiglottides were classified as leaf-shaped, curved (concaved or omega-shaped) and floppy types. We graded the airway opening at the level of epiglottis into six grades and obstruction at the tongue base level into four grades. The grades during inspiration (GrIns), expiration (GrExp) and after JTM (GrJTM) were recorded and compared with Pearson chi-square test. RESULTS: The strictly curved (Omega-shaped or concaved) epiglottis supplied a salvage pathway for airflow that resisted collapsing with the posterior movement of the tongue base in 2 patients. When we compared GrIns with GrExp for epiglottis the difference was statistically significant (chi(2) = 0.001), but the difference for tongue base was not (chi(2) = 0.152). After JTM, GrJTM for both epiglottis and tongue base were significantly better than GrIns and GrExp (chi(2) < 0.001). CONCLUSION: Tongue base was the principal site of obstruction although during the respiratory cycle the position of epiglottis changed prominently and increased the obstruction in inspiration. JTM alone significantly relieved the obstruction at the tongue base and epiglottis levels and increased the retroglossal airway. PMID- 15753622 TI - Regulation of apoptosis in external auditory canal cholesteatoma by hepatocyte growth factor/scatter factor. AB - OBJECTIVES: External auditory canal cholesteatomas (EACC) are characterized by focal invasion of squamous cell epithelium and accumulation of keratin debris in the apical part of the matrix. Apoptosis appears to be important in understanding the pathogenesis of EACC. Here the possible regulatory effect of the apoptosis mediated by hepatocyte growth factor (HGF)/scatter factor (SF)-c-Met-Fas in EACC is discussed. METHODS: We examined 17 EACC specimens for immunohistochemical expression of HGF/SF, c-Met, caspase 3 and Fas. The staining reaction was evaluated semiquantitatively. RESULTS: HGF/SF was detected in mesenchymal tissue below the EACC epithelium. c-Met was expressed throughout the epithelium. Fas and caspase 3 were detected at increasing levels towards the apical layers of the EACC matrix. CONCLUSIONS: High levels of HGF/SF result in binding of HGF/SF to c Met, releasing Fas to aggregate and bind to its death-inducing signaling complex. The result is apoptosis, marked by formation of dead squamous cells and sequestered keratin debris on the apical side of the cholesteatoma. PMID- 15753623 TI - Cerebrospinal fluid leaks of temporal bone origin: etiology and management. AB - BACKGROUND: Cerebrospinal fluid (CSF) leaks of the temporal bone region require surgical treatment as they pose life-threatening risks such as meningitis. AIM: The aim of the study was to determine the surgical outcome depending on different operation techniques and grafts. METHOD: We performed a retrospective review of 28 cases of CSF leaks, operated in our department from 1983 to 2002. After a mean follow-up of 8 years, patients were interviewed concerning otorrhea or rhinorrhea and meningitis. In this context, our management of CSF leaks is presented. RESULTS: The CSF leak had arisen spontaneously (n = 3), traumatically (n = 6) or postoperatively (n = 19). The surgical CSF leak repairs were performed via a transmastoid (n = 13), a middle fossa (n = 11) or a combined (n = 4) approach. Surgical outcome was independent on the used graft. CSF leak could be sealed in 25 of 28 cases. Only 3 patients suffered from recurrences. Meningitis or other complications did not occur. CONCLUSIONS: Comparing different techniques and grafts, there were no differences in the surgical outcome. PMID- 15753624 TI - A combination of budesonide and the SH-metabolite I of erdosteine acts synergistically in reducing chemiluminescence during human neutrophil respiratory burst. AB - Activated neutrophils can release superoxide anion and nitric oxide (NO), which subsequently combine with each other to yield peroxynitrite anions, powerful and harmful oxidants that preferentially mediate the oxidation of the thiol groups in proteins and non-protein molecules. These oxidants play a direct role in the inflammatory process in chronic obstructive pulmonary disease and asthma by increasing the number of neutrophils and macrophages that induce a self sustaining phlogogenic loop. Budesonide (BUD) and erdosteine (a muco-active drug which, after metabolization, produces an active metabolite (Met I) with a sulfhydryl group) are both active in reducing the release of superoxide anion, NO and peroxynitrite, and can be administered to patients with respiratory diseases. The aim of this study was to investigate the possible synergistic in vitro effect of BUD and Met I on chemiluminescence generation during fMLP-stimulated respiratory bursts of human neutrophils with the NO donor L-arginine, added to the incubating medium. The investigated BUD concentrations ranged from 6 x 10(-8) to 1 x 10(-6) mol/l in logarithmic scale and a significant and progressive reduction in luminol-amplified chemiluminescence (LACL) was observed at concentrations ranging from 2.5 x 10(-7) to 1 x 10(-6) mol/l. The investigated concentrations of Met I varied from 0.62 to 10 microg/ml. No significant changes were observed at 0.62, 1.25, and 2.5 microg/ml, but a significant decrease in LACL was observed at 5 and 10 microg/ml. When the two drugs were combined, there was a greater significant decrease in LACL versus the single drugs with the combinations of BUD 1 x 10(-6) mol/l plus Met I 10 microg/ml, BUD 5 x 10(-7) mol/l plus Met I 5 microg/ml, BUD 2.5 x 10(-7) mol/l plus Met I 2.5 microg/ml, and BUD 1.25 x 10(-7) mol/l plus Met I 1.25 microg/ml. A further interesting finding was that the combination of BUD 2.5 x 10(-7) mol/l plus Met I 2.5 microg/ml and BUD 1.25 x 10(-7) mol/l plus Met I 1.25 microg/ml significantly decreased LACL, whereas the single concentrations had no significant effect, thus indicating the possibility of extending the duration of the effect. Our findings indicate a synergistic antioxidant effect when BUD and Met I are given together, which is of interest for counteracting the airway phlogosis involved in many respiratory diseases. PMID- 15753626 TI - Unusual presentation of sarcoidosis. AB - The extrapulmonary presentation of sarcoidosis is a frequent occurrence - over 30% of cases when the observation is prospective - but early diagnosis may still be a challenge. A comparison between two different Italian series - (1) patients visiting the Milan Sarcoidosis Clinic of an Internal Medicine Department and (2) an Italian Register from many pneumology centers (RIPID) - has shown a different approach to the diagnosis because pneumologists are more likely to use transbronchial biopsy, while internists are more likely to use extrapulmonary biopsies. Such a different clinical approach may be due in part to a different case mix of the two series. PMID- 15753627 TI - Novel approaches to the treatment of bacterial lower respiratory tract infections: introduction. PMID- 15753628 TI - Strategic targets of essential host-pathogen interactions. AB - This review summarizes the present concepts regarding the biological processes that mediate intrinsic and innate host defense against microbial invasion of the lung. Innate immunity is the first line of defense of the higher organisms towards invading pathogens. It accomplishes a wide variety of activities including recognition and effector functions. The innate responses use phagocytic cells (macrophages, monocytes, and neutrophils), cells that release inflammatory mediators (basophils, mast cells, and eosinophils), and natural killer cells. The molecular component of innate responses includes complement, acute-phase proteins, and cytokines. Recognition of pathogen-associated molecular patterns is mediated by the pathogen receptors of the innate immune system, among these molecules toll-like receptors have emerged as fundamental components in the innate immune responses to infection, and a link between innate and adaptive immunity. Additional protection comes from polypeptide mediators of the innate host defense, such as the defensins and other antibiotic peptides. In view of the considerable burden in terms of mortality and morbidity that severe infections still pose worldwide, a better understanding of the biological basis of host pathogen interactions opens stimulating future treatment perspectives. PMID- 15753629 TI - Induced sputum: a new tool to monitor idiopathic pulmonary fibrosis? PMID- 15753630 TI - IL-18: a role in nonspecific interstitial pneumonitis pathogenesis and discrimination? PMID- 15753631 TI - Cockade-like structures in alveolar macrophages in extrinsic allergic alveolitis another piece in the puzzle? PMID- 15753632 TI - Induced sputum versus bronchoalveolar lavage fluid in the evaluation of patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: Induced sputum (IS) has been proposed as a useful noninvasive method for the assessment of airway diseases. Bronchoalveolar lavage fluid (BALF), an important tool for evaluating interstitial lung diseases, has limited utility due to its invasiveness and the difficulties of performing it in severely ill patients, while it is impractical for follow-up evaluation. OBJECTIVES: The aim of this study was to investigate the differences and the possible correlation of cell differential and lymphocyte subpopulations between BALF and IS samples in patients with idiopathic pulmonary fibrosis (IPF). METHODS: We studied prospectively 20 patients (18 male, 2 female) of median age 67 years (range 40 75) with IPF and 10 normal subjects (5 female, 5 male) of median age 59 years (range 36-70). IS was performed with hypertonic saline solution using an ultrasonic nebulizer (Ultraneb 2000). BALF was performed by a conventional procedure using fiberoptic bronchoscopy within 3 days from IS. May-Grunewald Giemsa-stained preps were differentially counted and T-lymphocyte subsets were analyzed by a flow-activated cell sorter. RESULTS: The percentage of macrophages was significantly lower in IS than in BALF (p < 0.0001), while the neutrophils were lower in BALF (p < 0.0001). A significant correlation was found between BALF and IS eosinophil counts (r = 0.54, p = 0.01) and CD4+/CD8+ ratio (r = 0.74, p < 0.0001). CONCLUSION: Our data suggest that different information is obtained by IS and BALF and thus, the two methods are complementary in IPF. PMID- 15753633 TI - Increased levels of interleukin-18 in bronchoalveolar lavage fluid of patients with idiopathic nonspecific interstitial pneumonia. AB - BACKGROUND: Interleukin-18 (IL-18) is a proinflammatory cytokine that can induce interferon-gamma (IFN-gamma), and it plays an important role in T-helper 1 responses. Among idiopathic interstitial pneumonia (IIP), nonspecific interstitial pneumonia (NSIP) has an increased number of lymphocytes in bronchoalveolar lavage (BAL) fluid compared with usual interstitial pneumonia (UIP). However, the difference in their pathogenesis is unclear. OBJECTIVE: The present study aims to investigate the roles of IL-18 in patients with idiopathic UIP and idiopathic NSIP. METHODS: We measured the serum and BAL fluid (BALF) levels of IL-18 and IFN-gamma in 22 patients with IIP (12 with UIP, 10 with NSIP) and 9 healthy volunteers. RESULTS: Lymphocyte proportions in BALF were significantly higher in NSIP than in UIP and healthy subjects. No significant differences were observed in the serum IL-18 levels of all subjects, while the BALF levels of IL-18 in patients with NSIP were significantly higher than in patients with UIP (p < 0.005) and in healthy subjects (p < 0.005). Among all subjects, the levels of IL-18 in BALF correlated significantly with those in serum and the lymphocyte proportions in BALF. The serum IFN-gamma levels of all subjects were below sensitivity, but there was significant reverse correlation between the levels of IFN-gamma and the lymphocyte proportions in BALF. CONCLUSION: The lymphocytosis in BALF of patients with idiopathic NSIP and a part of idiopathic UIP might be associated with the high levels of IL-18. PMID- 15753634 TI - Cockade-like structures in alveolar macrophages in extrinsic allergic alveolitis. AB - BACKGROUND AND OBJECTIVES: In immunocytochemical preparations of bronchoalveolar lavage (BAL) cells from patients with extrinsic allergic alveolitis (EAA), we observed the presence of alveolar macrophages with cockade-like structures in their cytoplasm (cockade+ alveolar macrophages). These cockade+ alveolar macrophages may reflect a subpopulation of alveolar macrophages which may show a different predominance in various interstitial lung diseases. In this study we aimed to compare the frequency of cockade+ alveolar macrophages in patients with EAA (n = 14) with the results obtained in patients with sarcoidosis (n = 11), idiopathic interstitial pneumonia (IIP; n = 10) and control subjects (n = 8). We also investigated the expression of the transferrin receptor CD71 on cockade+ alveolar macrophages. METHODS: In BAL fluid, the total number of cells and differential counts were determined, and immunocytologic examinations of macrophages and lymphocytes were done using monoclonal antibodies. The percentage of cockade+ alveolar macrophages was determined by counting 300 macrophages in the CD20 field of an immunocytochemical slide. RESULTS: The percentage of cockade+ alveolar macrophages was significantly higher in the EAA group (36 +/- 9%) compared to patients with sarcoidosis (12 +/- 5%) or IIP (11 +/- 10%) and control subjects (3 +/- 1%; p < 0.001). The proportion of CD71+ alveolar macrophages was significantly lower in EAA than in the other groups (p < 0.01), and the CD71 antigen was expressed on a significantly lower proportion of cockade+ alveolar macrophages compared to cockade- alveolar macrophages in EAA (p < 0.001). CONCLUSION: We conclude that cockade+ alveolar macrophages could play a role in the pathogenesis and differential diagnosis EAA. PMID- 15753635 TI - Questionnaire responses that predict airway response to hypertonic saline. AB - BACKGROUND: Airway hyperresponsiveness to hypertonic saline (HS) is associated with airway inflammation. We investigated if responsiveness to HS was predicted by asthma symptoms in the last 3 months. OBJECTIVES: To investigate if responsiveness to HS can be estimated by questionnaire items investigating asthma symptoms of the last 3 months. METHODS: Six hundred and four patients with physician-diagnosed asthma being assessed for asthma severity were studied. Bronchial provocation with 4.5% saline was performed, and a questionnaire was administered. The response to 4.5% saline was reported as the provoking dose to cause a 15% fall in the forced expiratory volume in 1 s FEV(1) (PD(15)) and the response-dose ratio (RDR). RESULTS: Based on the GINA guidelines, asthma severity was intermittent in 497 patients, mild in 107 patients, moderate in 3 patients and severe in 1 patient. A PD(15) to 4.5% saline was recorded in 234 of the 604. Questions on self-recognition of asthma, dust as a trigger, food as a trigger, and frequency of bronchodilator use were significant predictors for a PD(15), and currently taking steroids decreased the likelihood of a positive response to 4.5% saline. Using a multiple-linear regression model, a difference in the RDR could be calculated between those who answered positively compared with the reference group, who answered negatively. This difference could be used as a guide for predicting abnormal reactivity. An increase in RDR in response to 4.5% saline, compared with the reference group, was demonstrated in the presence of self recognition of asthma severity, dust and cats as a trigger or use of bronchodilator during sleep hours. CONCLUSIONS: Because of the high positive predictive value of HS for identifying patients with asthma it might be that the need for bronchodilator use at night not only predicts airway hyperresponsiveness to HS, it also could reflect the severity of asthma. PMID- 15753636 TI - Determinants of the length of mechanical ventilation in patients with COPD in the intensive care unit. AB - BACKGROUND: About 10% of the patients with chronic obstructive pulmonary disease (COPD) are at high risk for prolonged mechanical ventilation (MV >21 days), and mortality ranges from 55 to 78% in these patients. OBJECTIVE: To determine the potential risk factors for MV over periods of 1, 2 and 3 weeks in patients with COPD. PATIENTS AND METHOD: The characteristics of patients during the stable period of their disease, on admission to the intensive care unit (ICU) and during the ICU stay were recorded prospectively and analyzed retrospectively for this study. t test, chi(2) test and logistic regression analysis were used for statistical analysis. RESULTS: 86 patients with COPD requiring MV were included in the study. 73, 33, and 13% of the patients required MV longer than 1, 2 and 3 weeks, respectively. There were no significant relationships between the duration of MV and bronchiectasis or the presence of community-acquired pneumonia on admission, baseline pulmonary function test results or blood gas parameters on admission. Development of ventilator-associated pneumonia (VAP; odds ratio, OR: 6; 95% confidence interval, CI: 2-23, p = 0.011) and sepsis (OR: 10; 95% CI: 2 54, p = 0.007) were independent predictors for MV >7 days. VAP was still a risk factor for MV >15 days with an OR of 14 (95% CI: 3-66, p = 0.001). On the other hand MV >21 days was primarily determined by increasing age (OR: 1.2; 95% CI: 1 1.3, p = 0.042), severity of the disease on admission measured by APACHE II score (OR: 1.4; 95% CI: 1-1.7, p = 0.002) and albumin levels (OR: 0.10, 95% CI: 0.01 0.54, p = 0.007). CONCLUSION: Advanced age, severity of disease on admission and development of VAP during ICU stay are the main determinants of MV duration in patients with COPD. PMID- 15753637 TI - Role of cdk4, p16INK4, and Rb expression in the prognosis of bronchioloalveolar carcinomas. AB - BACKGROUND: The p16(INK4) protein has been identified as a potent inhibitor of cyclin-dependent kinase (cdk)4 by blocking cdk4-mediated phosphorylation of the tumor suppressor retinoblastoma (Rb) protein, thus allowing Rb-mediated growth suppression. OBJECTIVES: Loss of p16(INK4) has been associated with a poor cancer prognosis, but its potential significance in bronchioloalveolar carcinomas (BACs) has not been explored. METHODS: We examined immunohistochemical expression of p16(INK4), cdk4, and Rb proteins in 38 BACs and correlated their expression levels with known clinicopathological features of the disease. RESULTS: All BACs expressed cdk4, while 89 and 82% expressed p16(INK4) and Rb proteins, respectively. None of the clinicopathological factors correlated with p16(INK4), cdk4, or Rb expression separately. A low p16(INK4)/cdk4 ratio was significantly associated with a high disease stage (p = 0.04), and the ratio tended to be lower in mucinous than nonmucinous tumors. BACs with a low p16(INK4)/cdk4 ratio showed significantly higher Rb expression levels (p = 0.02). Univariable survival analyses showed a significantly lower 5-year survival probability in patients with a high stage (p = 0.002) or low p16(INK4)/cdk4 ratio (p = 0.01). CONCLUSIONS: The results suggest a role of the cdk4/p16(INK4) pathway in the prognosis of BACs. Further studies are warranted to clarify whether a low p16(INK4)/cdk4 ratio may identify tumors that are destined to behave unfavorably. PMID- 15753638 TI - Does 'idiopathic pleuritis' exist? Natural history of non-specific pleuritis diagnosed after thoracoscopy. AB - BACKGROUND: Even after a complete work-up including thoracoscopic biopsies, a significant number of patients with pleural exudates are diagnosed with 'non specific pleuritis', and no specific diagnosis can be made. The natural evolution of these patients is poorly understood. OBJECTIVES: To study the natural evolution of patients with non-specific pleuritis diagnosed after thoracoscopy and to evaluate whether the histological diagnosis of non-specific pleuritis corresponds with the clinical diagnosis of 'idiopathic pleuritis'. METHODS: We retrospectively studied the evolution of 75 patients between 1992 and 2002 (49 men and 26 women), mean (+/- SD) age 63.4 (+/- 13.3) years, who underwent diagnostic thoracoscopy because of an unexplained exudative pleural effusion, and in whom the histological diagnosis of non-specific pleuritis was made. Follow-up data were obtained through medical files and/or telephone contacts with general practitioners. RESULTS: Of these 75 patients, 8.3% eventually developed a malignancy during the follow-up period. In the remaining patients (91.7%), the clinical evolution followed a benign course. Ultimately, a probable cause was established on clinical grounds in 40 patients. True idiopathic pleuritis was finally observed in 25% of patients with the histological diagnosis of non specific pleuritis. Recurrence of the effusion occurred in 10 out of 60 (16.7%) patients, after a mean period of 26.2 months. CONCLUSIONS: The majority of non specific pleuritis patients (91.7%) followed a benign course, with a spontaneous resolution of the effusion in 81.8% of cases. In the majority of patients, a probable cause of the pleuritis was identified. True 'idiopathic benign pleuritis' hence occurs in only a minority (25%) of patients. PMID- 15753639 TI - Antioxidant responsiveness in BALB/c mice exposed to ozone. AB - BACKGROUND: A single, acute exposure to ozone has been shown to modify the antioxidant defense mechanism in the respiratory tract. OBJECTIVE: The aim of this study was to evaluate the effects of ozone exposure on antioxidant response in BALB/c mice. METHODS: We measured enhanced pause of breathing (Penh) as a marker of airway obstruction using barometric whole-body plethysmography before and after ozone exposure [groups (n = 6): filtered air, 0.12 ppm, 0.5 ppm, 1 ppm, 2 ppm] for 3 h. Antioxidant levels were measured using high-performance liquid chromatography with electrochemical detection in bronchoalveolar lavage (BAL) fluid and lung tissue homogenates. RESULTS: Malondialdehyde concentrations in lung tissue homogenates were significantly increased in the group exposed to 2 ppm ozone compared to the filtered air group. Uric acid and gamma-tocopherol concentrations in BAL fluid were significantly increased in the ozone exposure group compared to the filtered air group (p < 0.01). Uric acid concentrations were increased in a concentration-dependent manner according to ozone concentration to which the animals were exposed. Increases in Penh after ozone exposure were significantly higher in an ozone concentration-dependent manner. The proportion of neutrophils in BAL fluid was significantly higher in the group exposed to 2 ppm than in the filtered air and the group exposed to 0.12 ppm (p < 0.01, respectively). The level of ascorbate correlated with the level of gamma tocopherol. CONCLUSION: These findings suggest that antioxidant responses may serve as a protective mechanism against a range of oxidants in BALB/c mice exposed to ozone. PMID- 15753640 TI - Does rigid bronchoscopy induce bacterial translocation? An experimental study in rats. AB - BACKGROUND: Although some reports suggest that bronchoscopy induces bacterial translocation (BT), the mechanisms of BT remain unclear. OBJECTIVE: We aimed to assess whether bronchoscopy or hypoxemia during bronchoscopy is responsible for BT. METHODS: We evaluated 24 rats divided into three subgroups: the control group (group 1, n = 8); the rigid bronchoscopy group (group 2, n = 8), and the group receiving bronchoscopy + mechanical ventilation (group 3, n = 8). Oxygen saturation (SaO(2)) was measured during the bronchoscopic procedure. Blood and tissue cultures from mesenteric lymph nodes (MLNs), liver, spleen and cecal contents were obtained 24 h following bronchoscopy. RESULTS: In group 2, SaO(2) was significantly lower than in groups 1 and 3 (p < 0.01). In group 2, BT significantly increased (6/8, 75%; p < 0.01 vs. group 1, and p < 0.05 vs. group 3). The main site of translocation was MLNs (6/8, 75%) in group 2, while BT was detected in only 1 rat in group 3 (1/8, 12.5%). CONCLUSION: Hypoxemia during rigid bronchoscopy resulted in intestinal mucosal damage in a rat model. Hypoxemia may have been the trigger for BT from the intestine following bronchoscopy. PMID- 15753641 TI - Simultaneous existence of two different endobronchial disorders diagnosed by bronchoscopy. PMID- 15753642 TI - Pulmonary hypertension in men with thyrotoxicosis. AB - Thyrotoxicosis has a myriad of respiratory symptoms including dyspnea. Pulmonary hypertension may contribute to the respiratory symptoms of thyrotoxicosis, but is often unrecognized. We describe 3 male patients with thyrotoxicosis and associated pulmonary hypertension. Case reports of an additional 15 patients are also reviewed. In patients with thyrotoxicosis and pulmonary hypertension, treatment of thyrotoxicosis alone is associated with improvement in pulmonary hypertension. Previous reports have consisted of mostly female patients, but we report 3 men. When all cases are considered, the typical patient is female (10/14 = 71%), middle-aged (48 years), with mean pulmonary artery systolic pressures improving from 56 to 32 mm Hg with treatment. Autoantibodies were detected in 10/14 (71%) patients. The response to treatment (medical or surgical) of thyrotoxicosis supports the hypothesis that hyperthyroidism is either a cause of pulmonary hypertension, or a factor that may unmask pulmonary hypertension. Recognition is important since treatment and response are very different compared to other patients with pulmonary hypertension. This association may not be readily considered in men, since most reports have been of women. PMID- 15753644 TI - Bloodstained pleural effusion in a 38-year-old non-smoking female. PMID- 15753643 TI - Pulmonary malakoplakia coexistent with tuberculosis of the hilar lymph node mimicking malignancy. AB - Pulmonary malakoplakia is a disease that is difficult to diagnose because its symptoms vary from those mimicking bronchogenic carcinoma to pneumonitis-like tuberculous infections. Malakoplakia is an unusual inflammatory condition characterized by the accumulation of macrophages with diagnostic Michaelis Gutmann bodies. In this study, an immunocompromised patient with a tumor mass in the upper lobe of the right lung coexisting with hilar node enlargement is presented. A thoracoscopic biopsy revealed pulmonary malakoplakia with tuberculosis of the hilar lymph node. Microbiologic cultures were positive for acid-fast positive bacilli from the hilar node specimen, and negative for any other microorganisms such as Rhodococcus equi, but positive for Escherichia coli from the lung specimen. The significance of these findings and the coincidental association between malakoplakia and tuberculosis are discussed. PMID- 15753645 TI - Drugs in clinical development for chronic obstructive pulmonary disease. AB - Many drugs may be potentially useful in the treatment of chronic obstructive pulmonary disease (COPD), but relatively few become available for human use due to lack of safety, lack of efficacy, or both. This is an inherent risk in the drug development process, which coupled with the limited understanding of the molecular pathogenesis of COPD, has produced a trend toward improving existing compounds rather than to develop new compounds. This review focuses on improved existing compounds and newly discovered compounds that are in clinical trials, but not yet marketed. The improved existing compounds include: isomers of the long-acting bronchodilators, once-daily beta2-adrenoceptor agonists, anticholinergics and corticosteroids. The pool of novel compounds is in constant fluctuation and comprises anti-inflammatory drugs, antioxidants, leukotriene modifiers and a number of compounds aimed at treating different aspects of COPD such as pulmonary hypertension and hypophosphatemia. PMID- 15753646 TI - A terminological controversy: do disseminated and miliary tuberculosis mean the same? PMID- 15753647 TI - Telomerase RNA mutated in autosomal dyskeratosis congenita reconstitutes a weakly active telomerase enzyme defective in telomere elongation. AB - Dyskeratosis congenita (DC) is a rare multi-system syndrome characterized by nail dystrophy, abnormal skin pigmentation and mucosal leukoplakia. The gene mutated in the X-linked form of human DC encodes for dyskerin, a nucleolar pseudourydilase that is involved in rRNA maturation. Dyskerin is also involved in telomerase function through its interaction with the telomerase RNA (hTR). Mutations in dyskerin result in low levels of hTR, decreased telomerase activity and telomere shortening. Autosomal dominant DC is characterized by mutations in hTR, supporting the hypothesis that the DC phenotype may be caused by impaired telomere maintenance. Several mutations have been identified in different regions of hTR in patients affected by autosomal dominant DC. Recent reports have shown that coexpression of wild-type hTR with hTR harboring mutations found in the pseudoknot domain does not affect telomerase activity in vitro. However, these studies did not assess the consequences of mutant hTR expression at the telomeres. Here we provide the first direct in vivo evidence that a mutant hTR carrying the GC to AG double substitution in the pseudoknot at nucleotides 107 108 found in patients affected by autosomal dominant DC does not behave as a dominant-negative for telomere maintenance. Rather it reconstitutes a weakly active telomerase enzyme, which is defective in telomere elongation. PMID- 15753649 TI - BRAF kinase activation via chromosomal rearrangement in radiation-induced and sporadic thyroid cancer. AB - Activating point mutations of the BRAF gene have been recently described in a variety of human tumors. In a study published in the Journal of Clinical Investigation, we reported a novel mechanism of activation of this gene via paracentric inversion of chromosome 7q. The fusion protein, AKAP9-BRAF, contains the intact kinase domain and lacks the autoinhibitory N-terminal portion of BRAF. It exhibited constitutive activation of BRAF kinase and was transforming for NIH3T3 cells. This finding represents the first demonstration of RAF activation by chromosomal rearrangement in human tumors. AKAP9-BRAF was more common in radiation-induced thyroid tumors, whereas point mutations of BRAF predominated in sporadic tumors of the same type, demonstrating the association between environmental factors and specific mechanisms of BRAF activation. PMID- 15753650 TI - Analysis of chromosomes 9 and 11 aneuploidy frequency in pleural effusion of patients with and without malignancy: interphase FISH technique. AB - Fluids of body cavities result in a series of pathophysiological events associated with non-malignant and malignant conditions that lead to the formation of exudative effusion. Diagnosis of effusion from the patients is frequently troublesome for the cytologist because of the differentiation and biological behavior of different cells type in effusion. In the present study, chromosomal aneuploidy status in effusion cells derived from 32 patients including 14 patients with non-malignant and 18 patients with malignant diseases [including malign mesothelioma (n = 6), adeno carcinoma (n = 10), small cell carcinoma (n = 2)] was analyzed by using fluorescence in situ hybridization (FISH) with centromere specific probes for chromosomes 9 and 11. There was significant difference in the incidence of chromosomal 9 and 11 aneuploidies when compared with controls (P = 0.000). However, aneuploidies of chromosomes 9 and 11 in effusion cells from patients with malignant disease had significantly higher than in effusion cells from patients with non-malignant (P = 0.000), suggesting that chromosomes 9 and 11 are frequently involved in the status of disease. The present study indicates that there is a association between chromosomes aneuploidies and pleural effusion cell status. Chromosome aneuploidies in non malignant group may be an indicator of premalignancy. PMID- 15753651 TI - Cdk9 regulates neural differentiation and its expression correlates with the differentiation grade of neuroblastoma and PNET tumors. AB - Cdk9 is a member of the Cdc2-like family of kinases. Its cyclin partners are members of the family of cyclin T (T1, T2a and T2b) and cyclin K. The Cdk9/Cyclin T complex appears to be involved in regulating several physiological processes. Recently, Cdk9 has been identified as a regulator of the differentiation program of several cell types, such as muscle cells, monocytes and lymphocytes, suggesting that it may have a function in controlling specific differentiative pathways. We analyzed whether Cdk9 and Cyclin T1 may be involved in the regulation of neuron and astrocyte differentiation. Cdk9 and Cyclin T1 expression levels were monitored during the differentiation program of neuroblastoma and astrocytoma cell lines. Our results suggest that Cdk9/Cyclin T1 complex may be required for neuron differentiation induced by retinoic acid, because the expression level of the complex varies during differentiation, but no significant changes were observed in its expression in the astrocytoma cell line. In addition, the expression of Cdk9 and Cyclin T1 was evaluated by immunohistochemistry in samples of neuroblastoma, PNET (Primary Neuroectodermal Tumor) and astrocytoma tumors of different grades, in order to assess whether there was a correlation between Cdk9 expression and tumor grading. Our results show that in neuroblastoma and PNET tumor samples Cdk9 is more expressed the more differentiated the tumor is. Conversely, no significant alteration of Cdk9 expression was observed in astrocytoma tumor samples of different grades, thus confirming the results obtained for the cell lines. PMID- 15753652 TI - Mitotic spindle checkpoint inactivation by trichostatin a defines a mechanism for increasing cancer cell killing by microtubule-disrupting agents. AB - Microtubule-disrupting agents such as the taxanes comprise some of the most clinically useful chemotherapeutic agents and invoke the spindle checkpoint in proliferating cells. A robust spindle checkpoint in turn may forestall mitotic catastrophe, potentially providing a mechanism that permits cancer cells to survive transient exposure to these drugs. Previous reports on G2-M cell cycle progression by histone deacetylase inhibitors suggested a potential role in modulating the therapeutic efficacy of microtubule-disrupting agents. As both classes of agents are generally administered in clinical trials as pulse treatments, we investigated in human cancer cells the effects of brief treatments with the histone deacetylase inhibitor trichostatin A (TSA) alone or with nocodazole or paclitaxel (Taxol) on cell cycle progression and the spindle checkpoint. Treatment of synchronized cells with 200 ng/ml of TSA alone for eight hours to completely block class I and II HDACs did not interfere with progression into mitosis with chromosomal condensation as confirmed by MPM-2 expression. TSA treatment at this concentration surprisingly did not interfere with formation of the mitotic spindle or centrosomal separation, but instead led to missegregation of chromosomes, suggesting effects on the spindle checkpoint. Consistent with this hypothesis, TSA abrogated the phosphorylation and kinetochore localization of the mitotic checkpoint protein BubR1 and the phosphorylation of histone H3 after paclitaxel and nocodazole treatment. These effects in turn led to rapid cell death and considerably reduced clonogenic survival. These results together suggest that by inactivating the spindle checkpoint, TSA can potentiate the lethal effects of microtubule-disrupting drugs, a strategy that might be usefully exploited for optimizing anticancer therapy. PMID- 15753653 TI - Rare variant hypothesis for multifactorial inheritance: susceptibility to colorectal adenomas as a model. AB - The rare variant hypothesis postulates that genetic susceptibility to colorectal neoplasia within the general population is due to a number of low frequency variants in a variety of different genes. Each variant confers a moderate, but detectable, increase in relative risk of developing the disease. Recent evidence suggests that a quarter of patients with multiple adenomatous polyps are due to rare but functionally important variants in just five genes. PMID- 15753654 TI - High efficiency transduction of dendritic cells by adenoviral vectors targeted to DC-SIGN. AB - Dendritic cells (DCs) are a central element in the development of antigen specific immune responses. The lack of a specific and efficient technique for the in vivo delivery of antigens to DCs remains a major obstacle limiting a vaccine's ability to induce an effective immune response. The efficacy of adenoviral (Ad) vectors in this regard can be enhanced through alterations in vector tropism such that DC-targeted transduction is achieved. Here, the efficiency of DC transduction by Ad vectors retargeted to DC-specific ICAM-3 grabbing nonintegrin (DC-SIGN) was studied and compared to that of Ad vectors retargeted through CD40. A comparable and significant enhancement of gene transfer to monocyte derived DCs (MDDCs) was accomplished by means of an Ad vector harboring the Fc-binding domain of Staphylococcus aureus protein A in combination with antibodies to DC-SIGN or to CD40 or with fused complexes of human Ig-Fc with their natural ligands, i.e., ICAM-3 or CD40L, respectively. Whereas CD40-targeted Ad transduction resulted in a more profound phenotypic DC maturation, DC-SIGN- and CD40-targeted Ad both induced similar levels of IL-12 secretion. These data demonstrate the usefulness of DC-SIGN as a DC-restricted targeting motif for Ad-mediated vaccination strategies. PMID- 15753655 TI - Connecting reovirus oncolysis and Ras signaling. AB - Reovirus is a benign virus with innate oncolytic activity and is a potential novel therapeutic for a number of cancers. Reovirus can replicate in, and induce death of cancerous cells having an activated Ras pathway. Ras activation leads to the inactivation of cellular antiviral mechanisms, specifically removing the translation block on reovirus transcripts. This review outlines recent progress towards elucidating the molecular connection between the Ras-signaling pathway and reovirus replication. PMID- 15753656 TI - DNA repair in the context of chromatin. AB - Modulation of chromatin is essential to nuclear processes that utilize DNA, such as transcription, replication, and repair. For example, transcription is assisted by histone post-translational modifications, as well as chromatin-remodeling complexes, which alter the structure of chromatin. Furthermore, recent advancements in the fields of DNA repair and chromatin reveal that both histone modifications and chromatin-remodeling complexes are essential for the repair of DNA lesions. In particular, chromatin-modifying complexes, such as the INO80 chromatin-remodeling complex and the Tip60 histone acetyltransferase complex, associate with the DNA damage-induced phosphorylated H2AX, which is often referred to as gamma-H2AX. In S. cerevisiae, the association of INO80 with gamma H2AX is required for the recruitment of INO80 to sites of DNA double-strand breaks. Additionally, in Drosophila, Tip60 exchanges gamma-H2AX for unmodified H2A in regions of DNA damage. This report reviews recent studies that emphasize the intimate relationship between evolutionarily-conserved chromatin-modifying complexes and histone post-translational modifications in the repair of DNA damage. PMID- 15753657 TI - PTEN: a novel anti-oncogenic function independent of phosphatase activity. AB - The PTEN gene is an important tumor suppressor mutated in a number of cancers. To date, its growth regulatory properties have been intimately linked to its ability to act as a protein and phosphoinositol phosphatase. Inactivation of the enzymatic activity of PTEN is primarily due to direct mutation of its amino terminal catalytic domain but approximately 20% of mutations are in the carboxy terminus, which regulates membrane localization, protein stability, cellular migration and p53 function. We identified a novel protein that interacts with this domain, the v-jun transcriptional target, MSP58. Binding of MSP58 to PTEN results in the suppression of MSP58-mediated transformation. However, this PTEN effect does not require its catalytic activity, suggesting additional mechanisms of PTEN action. PMID- 15753658 TI - Chromatin remodeling by WSTF-ISWI at the replication site: opening a window of opportunity for epigenetic inheritance? AB - During DNA replication, chromatin states have to be accurately transmitted from the parental to the daughter strands for faithful epigenetic inheritance. Chromatin remodelling factors at the replication site are thought to be involved in this process. Recent work adds ATP-dependent nucleosome remodelling factors to this category of enzymes. The WICH complex, consisting of the ISWI-type ATPase SNF2H and the Williams Syndrome Transcription Factor (WSTF), binds to replication foci using PCNA, a key factor in DNA- and chromatin replication and DNA repair, as an interaction platform. Depletion of WSTF results in decreased chromatin accessibility, which is evident already in newly replicated DNA. This leads to heterochromatin formation on a global scale and a decrease in overall transcriptional activity. Here, we propose that WICH, by keeping nucleosomes mobile, provides access to the newly replicated DNA and may thereby create a window of opportunity after DNA replication for rebinding of factors that maintain the epigenetic state, and thus prevents aberrant heterochromatin formation. Our model may provide an explanation for the long-standing observation of a delay in chromatin "maturation" on newly replicated DNA, by connecting this delay with the action of PCNA-bound WSTF-ISWI, and highlights chromatin remodeling shortly after DNA replication as a critical point for regulation. PMID- 15753659 TI - Enhanced immune response to gastric cancer specific antigen Peptide by coencapsulation with CpG oligodeoxynucleotides in nanoemulsion. AB - CpG oligodeoxynucleotides (CpG ODN) have been shown to have potent adjuvant activity for a wide range of antigens. Of particular interest is their improved activity when closely associated with the antigen. The purpose of this study is to construct a nanovaccine coencapsulated with a gastric cancer specific antigen MG7 mimotope peptide and adjuvant CpG ODN 1645 using new nanotechnology as nanoemulsion and evaluate its immunocompetence. Nanoemulsion vaccine was prepared using magnetic ultrasound methods. BALB/c mice were immunized and the in vivo effectiveness was evaluated using tumor challenge assay. It was shown that the tumor masses formed in the mice immunized with coencapsulated nanovaccine (0.0825 g) markedly smaller (P < 0.01) than those formed in the mice immunized with nanovaccine encapsulated with antigen peptide alone (0.4465 g). A tumor inhibiting rate as high as 82.5% of the coencapsulated nanovaccine was obtained, while nanovaccine encapsulated with peptide only could not achieve the same effect (28.5%) (P < 0.01). Enzyme-linked immunospot assay (ELISPOT) showed that immunization using MG7 mimotope peptide coencapsulated with CpG ODN within the same nanoemulsion enhanced the frequency of splenocytes secreting IFN-gamma significantly (P < 0.01) when compared with immunization using MG7 peptide encapsulated in nanoemulsion alone (197spots/1 x 10(6) vs. 73 spots/1 x 10(6)). Cellular ELISA indicated that serum titer of antibody against MG7-Ag was significantly higher (P < 0.01) in mice immunized with coencapsulation form nanovaccine (0.7884) than that in the group immunized with nanovaccine encapsulated with MG7 peptide alone (0.3616). Using intracellular flow cytometric analysis, it was found that the IFN-gamma response was contributed by CD4+ T cells. Our experiments suggest that a vaccinal approach using nano-delivery system carrying in tumoral epitope and CpG ODN as adjuvant may have important implications for cancer therapy. PMID- 15753660 TI - Changing cell fate by nuclear reprogramming. AB - The differentiated state of somatic cells is remarkably stable, but nuclear transfer can efficiently override the stability of cell differentiation by nuclear reprogramming. Genes that were silent in the differentiated state are activated, whereas genes specific for the differentiated state are switched off during the reprogramming process. The epigenetic changes that occur after nuclear transfer to Xenopus oocytes involve chromatin remodelling and DNA demethylation. In particular, we have reported that reactivation of the mouse stem cell specific gene oct-4 depends on demethylation of CpG:s in the proximal oct-4 promoter. Here we discuss molecular mechanisms of nuclear reprogramming, with special emphasis on DNA demethylation. PMID- 15753661 TI - Mcl-1 regulates survival and sensitivity to diverse apoptotic stimuli in human non-small cell lung cancer cells. AB - Overexpression of anti-apoptotic Bcl-2 family members and deregulation of the pathways that regulate pro-apoptotic family members have been observed in non small cell lung cancers (NSCLC). Previous reports have identified both Bcl-2 and Bcl-x(L) proteins as survival factors in lung cancer cells since reductions in these proteins can induce apoptosis and sensitize lung cancer cells to apoptosis induced by chemotherapy agents. Myeloid cell leukemia-1 (Mcl-1), another member of the Bcl-2 family, has been found to be a critical survival factor in hematopoietic cells, yet little data exists for a role of Mcl-1 in human lung cancers. We used NSCLC cell lines to explore how Mcl-1 levels affect lung cancer cell survival and studied tumors from patients to determine expression patterns of Mcl-1. NSCLC cells express abundant Mcl-1 protein and depletion of Mcl-1 levels by antisense Mcl-1 oligonucleotides induces apoptosis in A549 and H1299 lung cancer cells. Reduction in Mcl-1 levels can sensitize lung cancer cells to apoptosis induced by cytotoxic agents as well as by ionizing radiation. Lung cancer cells overexpressing Mcl-1 are less sensitive to apoptosis induced by chemotherapeutic agents, ZD1839 (an inhibitor of EGFR tyrosine kinase) and Bcl-2 or Bcl-x(L) antisense oligonucleotides. We find that epidermal growth factor (EGF) can enhance Mcl-1 protein levels in an ERK-dependent manner. Signal transduction agents that reduce Mcl-1 levels correlated with their individual ability to induce apoptosis in lung cancer cells. Finally, NSCLC tumors taken directly from patients have elevated levels of Mcl-1 protein compared with normal adjacent lung tissue. Therefore, agents that target Mcl-1 can induce apoptosis and sensitize cells to apoptosis induced by cytotoxic agents. Mcl-1 protein is overexpressed in a subset of human NSCLC and enhanced levels of Mcl-1 may protect lung cancer cells from death induced by a variety of pro-apoptotic stimuli. PMID- 15753662 TI - DNA damage-induced cohesion. AB - The protein complex Cohesin, forming protein-links that hold sister chromatids together, is at the heart of chromatid cohesion. Cohesion is important both for correct chromosome segregation and double-strand break (DSB) repair, making Cohesin central for the maintenance of genome stability. Until now, establishment of Cohesin links between chromatids has been shown to occur during DNA replication only. Recently it was however observed that in cells arrested in G2/M, DSB induction not only elicits chromosomal recruitment of Cohesin, but also formation of chromatid cohesion. The establishment of cohesion outside the period of replication opens a new field of investigation. Here we present results further supporting the formation of sister chromatid cohesion in response to DNA damage, and propose a model of how damage-induced cohesion could contribute to the linkage of chromatids during normal cell cycle progression. PMID- 15753664 TI - Experiencing the Athens 2004 Olympic Games at the polyclinic of the Olympic Village. PMID- 15753663 TI - The methods for determining the purity and in vitro or in vivo activity of recombinant human endostatin. AB - In order to establish the methods of high-performance liquid chromatography (HPLC) for determining the purity of recombinant human endostatin (rhEndostatin) and in vitro or in vivo activity of rhEndostatin, two columns were firstly used in HPLC analysis for determining the purity of rhEndostatin, including Waters Symmetry 300C4 (4.6 mm x 250 mm, 5 microm) and the Superdex75 HR 10/30. Cell lines, bovine capillary endothelial cells (BCEs) or human umbilical vein endothelial cells (HUVECs) expression human vascular endothelial growth factor (hVEGF) were used in method MTT or LDH as substrate, respectively. The bioactivity in vivo was assayed by the anti-tumor proliferation rate in H22 liver tumor-bearing mice. The results showed that the retention time of rhEndostatin sample was stable at 19.066 min or 11.506 min in reverse phase HPLC (RP-HPLC) or gel filtering HPLC (GF-HPLC). The stableness, repeat and recovery rates were over 99% in both methods and there was no statistical difference between these two methods (p > 0.05). In nonserum culture medium, rhEndostatin can sensitively and stably inhibit the proliferation of the HUVEC cells that were transfected with plasmid encoding hVEGF. LDH substrate methods is the most sensitive and stable method. The anti-tumor activity in H22 tumor-bearing mice was also highly repeatable and had an inhibition rate over 50% at 20 mg kg(-1) weight. As a conclusion, the RP-HPLC and GF-HPLC set up in this paper are highly repeatable, accurate and sensitive for detecting the purity of rhEndostatin. The bioactivity of rhEndostatin can be measured through detection the proliferation-inhibition on HUVECs transfectants with hVEGF in vitro or on H22 liver tumor in vivo. PMID- 15753665 TI - A pilot case-control study of gamma-radiation sensitivity and risk of papillary thyroid cancer. AB - In vitro gamma-radiation-induced chromatid breaks per cell (b/c) in lymphocytes may be associated with risk of papillary thyroid cancer (PTC). This pilot case control study involved 106 patients with thyroid disease (57 with PTC and 49 with benign thyroid disease) and 105 cancer-free matched controls. Multivariate logistic regression analyses identified that an elevated gamma-radiation-induced b/c value was a risk factor for PTC (adjusted odds ratio = 4.54; 95% CI, 2.07 9.95), and a dose-response relationship was evident when the b/c values were categorized into tertiles. High levels of chromatid breaks induced by gamma radiation may constitute an independent risk factor for PTC, but further study is needed. PMID- 15753666 TI - Investigation of loss of heterozygosity and SNP frequencies in the RET gene in papillary thyroid carcinoma. AB - In both medullary carcinoma and papillary carcinoma of the thyroid, altered expression of the RET gene is implicated in tumorigenesis. Recent studies suggest that loss of heterozygosity (LOH) at the G691S SNP may be associated with tumors from patients with a history of radiation exposure. We investigated LOH for three RET SNPs (G691S, S904S, and L769L) in tumor and normal tissue from 46 patients from Ukraine and Belarus who were exposed to radioactive fallout following the Chernobyl nuclear accident and were operated for papillary thyroid carcinoma between 1995 and 2000. Normal tissue from 28 patients was heterozygous for at least one SNP; DNA from the corresponding tumor samples was also heterozygous, indicating that no LOH had taken place. To assess SNP frequencies in a radiation associated thyroid cancer cohort, we investigated a further 68 unpaired post Chernobyl samples. For G691S, there was considerable deviation from Hardy Weinberg equilibrium; more detailed analysis showed that this was linked to age at onset of disease. Among younger patients, the distribution of genotypes conformed to Hardy-Weinberg equilibrium; among older patients, we observed marked deviation (p = 0.0072), with significant over-representation of the rare S allele relative to the younger groups (Fisher's exact, p = 0.0233). This suggests that SNPs in the RET oncogene may play a role in sporadic papillary thyroid carcinoma. PMID- 15753667 TI - Thyroid cancers express CD-40 and CD-40 ligand: cancers that express CD-40 ligand may have a greater risk of recurrence in young patients. AB - The immune response might suppress thyroid cancer recurrence. Although the factors that control this are unknown, CD-40 and CD-40 ligand might be important. To test this, we stained 36 papillary (PTC) and four follicular (FTC) thyroid carcinomas for CD-40 (n = 37) and CD-40 ligand (n = 36) and graded staining from absent (grade 0) to intense (grade 3). Follicular cells of the majority of thyroid tumors expressed CD-40 (30/37, 81%) and CD-40 ligand (15/24, 69%). Cancers from young patients (< or =21 years of age) that expressed CD-40 contained more numerous lymphocytes/high-power field (36 +/- 11) than cancers that failed to express CD-40 (4 +/- 3, p = 0.01), but there was no correlation with clinical outcome. Among young patients, CD-40 ligand expression was more intense in multifocal (1.1 +/- 0.2 vs. 0.45 +/- 0.2, p = 0.037), aggressive (1.14 +/- 0.14 vs. 0.65 +/- 0.2, p = 0.05) and recurrent tumors (1.2 +/- 0.2 vs. 0.65 +/- 0.2, p = 0.05) and associated with reduced disease-free survival (p = 0.03). We conclude that the majority of thyroid cancers express CD-40 and CD-40 ligand. In patients < or =21 years of age, tumors with intense expression of CD-40 ligand are more often multifocal, aggressive, and recurrent. PMID- 15753669 TI - Karl Hurthle! Now, who was he? AB - The name Hurthle is used in medicine to designate a particular type of thyroid cell, mainly characterized by a granular and deeply eosinophilic cytoplasm. This article describes the life of Karl Hurthle (1860-1945) and the experiments he performed in the early 1890s to explore the morphology and function of the thyroid gland. PMID- 15753668 TI - Thyrotropin-stimulated DNA synthesis and thyroglobulin expression in normal and hyperthyroid feline thyrocytes in monolayer culture. AB - Feline hyperthyroidism is a common, spontaneous disease in older cats that is similar clinically and histopathologically to human toxic multinodular goiter (TNG). In this study, the functional response of feline normal thyroid (NT) and hyperthyroid (HT) cells grown in monolayer culture to thyrotropin (TSH) was determined. Basal levels of DNA synthesis were similar in NT and HT cells. TSH stimulated concentration-dependent DNA synthesis in NT and HT cells, with maximal stimulation seen at 1 and 10 mU/mL TSH in NT and HT cells, respectively. HT cells had higher basal levels of thyroglobulin (Tg) expression. TSH stimulated Tg expression in NT and HT cells in a concentration-dependent fashion, with maximal activity at 0.5 and 5 mU/mL TSH, respectively. These results demonstrate that NT and HT cells in monolayer culture exhibit growth and functional responses to TSH. HT cells have higher basal Tg expression than NT cells and require higher TSH concentrations to stimulate DNA synthesis and Tg expression, two measures of thyroid cell activation. These data support the idea that feline hyperthyroidism is caused by cell abnormalities, resulting in dysregulated growth and hormone synthesis, and emphasize its importance as an animal model for TNG. PMID- 15753670 TI - Which thyroxine? PMID- 15753671 TI - Ultrasound-detected thyroid nodules in radiation-exposed patients: changes over time. AB - The relationship between radiation exposure and thyroid cancer is well known, but whether all irradiated patients should have thyroid ultrasounds is unresolved. We have performed follow-up ultrasound examinations of patients in a cohort who were exposed to conventional external radiation during 1939-63 for benign conditions of the head and neck area prior to their 16th birthday. Of 54 subjects who had normal radionuclide scans in 1974-76 and were reexamined in 1996-97 by thyroid ultrasonography, 42 remained eligible and 34 agreed to participate in the present ultrasound study. After an additional 4-8 years of follow-up and using an ultrasound machine with increased resolution, we found 160 nodules (in 33 of these 34 subjects), compared with 96 nodules (in 29 of the 34 subjects) detected in the previous examination. Only four of the new nodules were > or =10 mm. Of the previously diagnosed large (> or =10 mm) nodules, four nodules in four subjects resolved; nine nodules in six subjects regressed to <10 mm; 14 nodules in 13 subjects remained at > or =10 mm. The four new large nodules appeared in four subjects, and six small nodules increased to > or =10 mm in six other subjects. The total volume of the thyroid nodules decreased in the 13 subjects on thyroid hormone (by 0.20 cm(3)) and increased in the 21 subjects who were not (by 0.34 cm(3), p < 0.05 by unpaired t-test). In summary, thyroid nodules are extremely common in irradiated subjects. Many new ones may be observed over time, but most are small and seen because of the increased resolution of ultrasound machines. Compared to patients on no medication, nodules in patients on thyroid hormone tended to regress. Since FNA of all thyroid nodules in irradiated patients is not feasible, ultrasound is useful in identifying those lesions that are growing. PMID- 15753672 TI - Human recombinant TSH preceding a therapeutic dose of radioiodine for multinodular goiters has no significant effect in the surge of TSH-receptor and TPO antibodies. AB - Radioiodine (RAI) treatment has increasingly been used mostly in elderly patients with multinodular goiter (MNG) as an alternative for surgery. Recombinant human thyrotropin (rhTSH) has been demonstrated to increase the uptake of RAI and also to promote a more even distribution of radionuclide among the various nodules. We have compared the surge of autoantibodies to thyroid peroxidase (anti-TPO) and to the TSH receptor (TRAb) in two groups of patients with MNG. Group RAI (n = 15) received only RAI, and Group RAI+rhTSH (n = 15) received RAI 24 h after 0.45 mg of rhTSH intramuscularly. At baseline, all 30 patients had negative anti-TPO antibodies. After RAI, 16 patients (eight in each group) exhibited a positive anti-TPO test (range, 70-2359 U/mL). In the rhTSH-treated group, anti-TPO values were significantly higher (as compared to basal levels; p < 0.02) after 3 months of RAI treatment. After 12 months, the anti-TPO values decreased to lower but still positive concentrations in nine patients (Group RAI: three patients; Group RAI+rhTSH: five patients). Only one patient had a positive TRAb test at baseline (67.5% inhibition of the TSH binding). After RAI, positive TRAb values were present in 21/30 patients. After 6 months of RAI treatment, there was a significant increase of the TRAb values in Group RAI+rhTSH patients. After 12 months, only four patients had positive TRAb (Group RAI: three patients; Group RAI+rhTSH: one patient). Two patients, one of each group, had an elevation of free T4 levels and suppressed serum TSH values, indicating hyperthyroidism (Graves' disease). Bioassay of TSH receptor (TSHR) indicated absence of a significant elevation of cAMP in the medium before and after RAI treatment in all patients. Moreover, predominantly blocking TSHR autoantibodies were detected in six of the 30 patients (three of each group). Sera from these patients were able to reduce the TSH-stimulated cAMP generation by CHO cells. We conclude that the autoantibodies to the TSHR and to TPO may occur after RAI treatment of patients, either with or without previous stimulation by rhTSH. The antibodies to the TSH comprised a combination of agonist (stimulating) and antagonist (blocking) antibodies, which in most patients did not induce clinical and laboratory evidence of active Graves' disease. PMID- 15753673 TI - Inherited medullary thyroid cancer and the duty to warn: revisiting Pate v. Threlkel in light of HIPAA. AB - Familial medullary thyroid cancer (FMTC) is one of the few autosomal dominant cancers for which genetic testing provides a clear medical indication for prophylactic and/or curative therapy, and for which prophylactic thyroidectomy, followed by thyroid hormone replacement, presents a relatively low morbidity risk. Medullary thyroid cancer (MTC) is a particularly aggressive type of thyroid cancer, and screening by traditional biochemical markers yields a high proportion of advanced stage diagnoses in individuals from FMTC families. This is particularly hazardous since there are no curative systemic treatments for MTC. Genetic testing for germline mutations of the RET proto-oncogene provides a reliable method of identifying at-risk family members in those FMTC families in which a mutation has been identified in the proband. Prophylactic thyroidectomy in such at-risk family members has significantly reduced the proportion of advanced stage MTC diagnoses in MTC families. Since a clear medical benefit exists for genetic testing in family members, and a clear danger to family members exists in the absence of genetic counseling, establishing genetic diagnosis as standard of care has critical legal and ethical implications for medical providers caring for probands and family members. The "duty to warn," reinforced by the courts in the legal case of Pate v. Threlkel, may override recent confidentiality legislation, known as the HIPAA Privacy Rules, which came into effect April 12, 2003. PMID- 15753674 TI - Volume changes in intra- and extraorbital compartments in patients with Graves' ophthalmopathy: effect of smoking. AB - The objective of this study was to demonstrate the effect of smoking history on soft tissue expansion in specific orbital compartments in patients with Graves' ophthalmopathy. The volumes of the rectus muscles, intra and extraorbital connective, and soft tissues were measured in 110 orbits of 35 patients and 20 control subjects. Data sets from current smokers, ex-smokers, and non-smokers were compared. The total number of cigarettes smoked was calculated, and it was used as an estimate for the severity of smoking (cumulative smoking). The volume measurements were performed on T1-weighted contiguous transversal magnetic resonance images of the orbits. Connective tissue volumes were influenced by smoking history, while muscle volumes were not affected. Ex-smokers had larger amount of extraorbital connective tissue than current smokers (p = 0.012), and this volume showed a good correlation with the number of cigarettes smoked (r = 0.539, p < 0.05). In current smokers, the amount of intraorbital connective tissue correlated well with the cumulative smoking (r = 0.635, p < 0.001). We conclude that connective tissue volumes in certain orbital compartments correlate well with cumulative smoking. Extraocular muscle volumes are not influenced by smoking in patients with Graves' ophthalmopathy. PMID- 15753675 TI - Epidemiology of thyroid microcarcinoma found in autopsy series conducted in areas of different iodine intake. AB - The prevalence of thyroid microcarcinomas found at autopsies is 100-1000 times higher than in clinical cancer. The epidemiological and histological characteristics of thyroid microcarcinomas in consecutive series of autopsies performed in two areas of different iodine intake were investigated. Iodine deficient (ID) area: n = 222 (M = 109, F = 113), median age: 74-76 years, median iodine excretion (MIE) of nursing home residents from this area: 70 microg/g creatinine. Iodine sufficient (IS) area: n = 221 (M = 132, F = 89), median age: 68 years, MIE: 500 microg/g creatinine. When compared to the IS area, the results obtained in the ID area showed a higher thyroid weight (mean 27.75 g +/- 18.43 g vs. 16.5 g +/- 9.6 g, p < 0.0001) and a larger number of goitrous glands (50/222 vs. 5/221, p < 0.0001). Altogether 21 microcarcinomas were found (4.74%) with no iodine intake- or gender-related difference: ID n = 11 (4.95%), M/F = 8/3; IS n = 10 (4.52%), M/F = 6/4. Microcarcinomas seemed to be more prevalent in the 40-59 year age group. All microcarcinomas were of the papillary type. In conclusion, compared to clinical cancer, thyroid microcarcinomas are characterized by a two scale higher prevalence, are not related to iodine intake, gender or nodularity, are most exclusively of the papillary type. PMID- 15753677 TI - High prevalence of extrathyroid malformations in a cohort of Brazilian patients with permanent primary congenital hypothyroidism. AB - The objective of this study was to evaluate the prevalence of major congenital malformations in a cohort of children with permanent primary congenital hypothyroidism (PCH). This cross-sectional study was conducted in 76 consecutive, unselected children with PCH recruited from a district hospital outpatient clinic. Malformations were identified by clinical examination. The prevalence of major congenital malformations in these patients was compared with the prevalence of malformations in children born at Hospital de Clinicas de Porto Alegre (HCPA). The etiology of hypothyroidism was established by scintigraphy and ultrasonography. Hypothyroidism was caused by thyroid dysgenesis in 67 patients (one case of hemiagenesis, 24 of ectopia, and 42 of agenesis); the gland was normally located and shaped in nine patients. Ten patients (13.2%) had major congenital malformations (1,316/10,000 patients), mostly cardiac. Malformations were observed only in patients with dysgenesis: thyroid agenesis (n = 7) and sub lingual ectopic thyroid (n = 3). The prevalence of malformations was significantly higher (RR = 2.6; CI 95%: 1.3-4.8; p = 0.005) in this group than in HCPA newborns (509/10,000 patients). In conclusion, a high rate of extrathyroid congenital malformations, mostly cardiac, was found for patients with permanent PCH, especially thyroid dysgenesis. The present data support the existence of an association between CH and increased prevalence of congenital malformations. PMID- 15753676 TI - Clinical value of different responses of serum thyroglobulin to recombinant human thyrotropin in the follow-up of patients with differentiated thyroid carcinoma. AB - In the present study, we examined the clinical value of a differential response of thyroglobulin (Tg) concentration after recombinant human thyrotropin (rhTSH) stimulation (rhTSH Tg testing) and its correlation with (131)I uptake and whole body scanning (rhTSH-WBS) in 104 patients who had previously undergone near total thyroidectomy and (131)I ablation for differentiated thyroid carcinoma (DTC). RhTSH Tg testing was considered negative for rhTSH-Tg < 0.9 ng/mL, low positive for rhTSH-Tg of 1-5 ng/mL and high positive for rhTSHTg > 5 ng/mL. RhTSH Tg testing was negative in 70 patients, one of whom had a lymph-node metastasis, but no (131)I uptake. Seven patients had low positive rhTSH Tg testing and no (131)I uptake, but two of these patients had cervical lymph-node metastases. Twenty seven patients had high positive rhTSH Tg testing and (131)I uptake was detected in lung, bone, or mediastinum in 11. Imaging techniques (CT, MRI, FDG-PET) documented metastatic disease in 22. In conclusion, our results suggest that any rise in rhTSH-Tg, even at low level, should raise the suspicion of persistent or recurrent DTC. Patients with rhTSH-Tg at high level should be carefully evaluated, since DTC persistence is highly probable. TSH-WBS provides little adjunctive information. PMID- 15753678 TI - Littoral cell angioma presenting as metastatic thyroid carcinoma to the spleen. AB - Papillary thyroid carcinoma (PTC) commonly metastasizes to cervical lymph nodes. Distant metastases are unusual with the lungs most frequently involved. Well differentiated thyroid carcinoma very rarely presents with metastases to the spleen. This is the case of a 25-year-old man with a history of PTC (1.4 cm primary; no capsular invasion and negative lymph node metastases). One year after initial surgery, recurrent disease was found in multiple neck nodes by central neck dissection. Whole body scan (WBS) following a therapeutic ablation dose of 150 mCi I(131) revealed mediastinal metastases. Computerized axial tomography (CT) of the chest one year later showed no gross mediastinal or pulmonary disease. However, multiple large splenic lesions were incidentally noted. Evaluation by ultrasound (US) showed lesions to be solid echogenic masses without remarkable Doppler characteristics to suggest vascular tumors. US-guided percutaneous fine-needle aspiration biopsy (FNAB) of one lesion was nondiagnostic. After withdrawal from Levothyroxine, serum TSH was >100 mU/L with a thyroglobulin of 9.4 ng/mL and negative anti-thyroglobulin antibodies. Diagnostic WBS revealed faint splenic uptake but was otherwise unremarkable. Following treatment with 192 mCi I(131), WBS demonstrated increased activity in the mediastinum as well as in the spleen suggesting mediastinal and splenic metastases. Contrast CT of the abdomen showed multiple low-attenuated heterogeneously enhancing splenic masses, normal liver and no intra-abdominal lymphadenopathy. The largest mass (4.5 x 3.5 cm) was exophytic and in close proximity to the splenic capsule. Despite the serum thyroglobulin of only 9.4 ng/mL, the finding of I(131) accumulation within solid splenic masses led to a preoperative diagnosis of thyroid carcinoma metastases. To establish the diagnosis and to remove the risk for splenic rupture, a laparoscopic splenectomy was performed. Histopathologic analysis showed large littoral cell angiomas (LCA). False-positive radioiodine scintigraphy in the setting of PTC involving a vertebral hemangioma has been reported. To our knowledge, this is the first case that describes multiple angiomas mimicking metastatic thyroid carcinoma to the spleen. In one-third of all cases reported, LCA co-exists with various visceral organ cancers or malignant lymphoma. This is the first report of an association between LCA and thyroid carcinoma. PMID- 15753679 TI - Thyroid metastasis of pulmonary adenocarcinoma. PMID- 15753680 TI - Discrete nodule on ultrasound of the thyroid gland. PMID- 15753715 TI - High-frequency oscillatory ventilation for adult acute respiratory distress syndrome: a decade of progress. PMID- 15753716 TI - High-frequency oscillatory ventilation: lessons from the neonatal/pediatric experience. AB - Efforts to minimize ventilator-induced lung injury in adults with hypoxemic respiratory failure have recently focused on the potential role of high-frequency oscillatory ventilation (HFOV). However, HFOV has been studied in newborns with hypoxemic respiratory failure for nearly 3 decades. In this brief review, we attempt to summarize key physiological principles learned from this cumulative neonatal/pediatric experience with HFOV. PMID- 15753717 TI - Dynamic alveolar mechanics and ventilator-induced lung injury. AB - OBJECTIVES: To review the mechanism of dynamic alveolar mechanics (i.e., the dynamic change in alveolar size and shape during ventilation) in both the normal and acutely injured lung; to investigate the alteration in alveolar mechanics secondary to acute lung injury as a mechanism of ventilator-induced lung injury (VILI); and to examine the hypothesis that the reduced morbidity and mortality associated with protective strategies of mechanical ventilation is related to the normalization of alveolar mechanics. DATA EXTRACTION AND SYNTHESIS: This review is based on original published articles and review papers dealing with the mechanism of lung volume change at the alveolar level and the role of altered alveolar mechanics as a mechanism of VILI. In addition, data from our laboratory directly visualizing dynamic alveolar mechanics is reviewed and related to the literature. CONCLUSIONS: The mechanism of alveolar inflation in normal lungs is unclear. Nonetheless, normal alveoli are very stable and change size very little with ventilation. Acute lung injury causes marked destabilization of individual alveoli. Alveolar instability causes pulmonary damage and is believed to be a major component in the mechanism of VILI. Ventilator strategies that reduce alveolar instability may potentially reduce the morbidity and mortality associated with VILI. PMID- 15753718 TI - High-frequency oscillatory ventilation and ventilator-induced lung injury. AB - INTRODUCTION: Although mechanical ventilation is lifesaving for patients with acute respiratory distress syndrome, it can cause ventilator-induced lung injury. To minimize ventilator-induced lung injury, different ventilatory strategies have been developed. One of the strategies is the use of high-frequency oscillatory ventilation (HFOV). THEORETICAL BACKGROUNDS OF VENTILATOR-INDUCED LUNG INJURY AND HFOV: Because of the novel gas exchange mechanisms, HFOV can provide adequate gas exchange using extremely small tidal volumes and maintain high end-expiratory lung volume without inducing overdistension, which should result in minimization of ventilator-induced lung injury. STUDIES OF HFOV AND LUNG INJURY: There are convincing clinical and animal data indicating that HFOV is an ideal lung protective ventilatory strategy, particularly in the setting of neonatal respiratory failure, if lung volume recruitment is performed. CLINICAL IMPLICATION OF HFOV IN ADULT ACUTE RESPIRATORY DISTRESS SYNDROME: A recent clinical trial demonstrated early (<16 hrs) improvement in oxygenation with HFOV and a 30-day mortality of 37% with HFOV vs. 52% with pressure-controlled ventilation (p = .102), suggesting that HFOV is as effective and safe as the conventional strategy in adult acute respiratory distress syndrome. Future studies examining optimal algorithms of HFOV using clinically relevant animal models, and patients with acute respiratory distress syndrome, are imperative to determine whether the wide-spread application of HFOV is warranted in adult acute respiratory distress syndrome. PMID- 15753719 TI - High-frequency oscillatory ventilation: mechanisms of gas exchange and lung mechanics. AB - OBJECTIVE: Overview of the mechanisms governing gas transport, mechanical factors influencing the transmission of pressure and flow to the lung, and the measurement of lung mechanics during high-frequency oscillatory ventilation (HFOV) in acute respiratory distress syndrome. DATA SOURCES AND STUDY SELECTION: Studies indexed in PubMed illustrating key concepts relevant to the manuscript objectives. Pressure transmission during HFOV in the adult lung was simulated using a published theoretical model. DATA SYNTHESIS: Gas transport during HFOV is complex and involves a range of different mechanisms, including bulk convection, turbulence, asymmetric velocity profiles, pendelluft, cardiogenic mixing, laminar flow with Taylor dispersion, collateral ventilation, and molecular diffusion. Except for molecular diffusion, each mechanism involves generation of convective fluid motion, and is influenced by the mechanical characteristics of the intubated respiratory system and the ventilatory settings. These factors have important consequences for the damping of the oscillatory pressure waveform and the drop in mean pressure from the airway opening to the lung. New techniques enabling partitioning of airway and tissue properties are being developed for measurement of lung mechanics during HFOV. CONCLUSIONS: Awareness of the different mechanisms governing gas transport and the prevailing lung mechanics during HFOV represents essential background for the physician planning to use this mode of ventilation in the adult patient. Monitoring of lung volume, respiratory mechanics, and ventilation homogeneity may facilitate individual optimization of HFOV ventilatory settings in the future. PMID- 15753720 TI - High-frequency oscillatory ventilation: lessons learned from mechanical test lung models. AB - OBJECTIVE: Review data obtained from high-frequency oscillatory ventilation (HFOV) and mechanical test lung models with respect to delivered tidal volume, distal pressure transmission, endotracheal tube cuff leaks, and simulated clinical conditions. DESIGN: Review of selected studies from PubMed, published abstracts, and institutional mechanical test lung data. RESULTS: Tidal volume delivery during HFOV is altered by oscillatory pressure amplitude (DeltaP), frequency (Hz), percent inspiratory time (IT%), and patient variables. Distal (carinal) oscillatory pressure amplitude transmission is directly correlated with endotracheal tube diameter and peripheral airway resistance. Endotracheal tube cuff leaks promote egress of tracheal gas while attenuating distal oscillatory pressure amplitude and tidal volume transmission. Simulated clinical conditions (e.g., increased distal airway resistance, mainstem intubation) may increase observed DeltaP, whereas mean airway pressure is decreased with air leaks. CONCLUSION: Mechanical test lung and artificial trachea simulations may provide useful information on the interaction of HFOV with altered lung mechanics and may contribute to the formulation of HFOV clinical strategies. Important limitations of these models include absence of gas exchange, histologic and biologic markers, or hemodynamic data. PMID- 15753722 TI - Computed tomography scan assessment of lung volume and recruitment during high frequency oscillatory ventilation. AB - OBJECTIVE: This review describes how computed tomography has increased our understanding of the pathophysiology of acute respiratory distress syndrome. It summarizes current knowledge about lung volume changes and alveolar recruitment during high-frequency oscillatory ventilation (HFOV) assessed by computed tomography (CT), outlines potential problems when comparing HFOV with conventional ventilation (CV) as a result of the different pressure-time profiles, and describes future research directions. DATA SOURCE: CT allows accurate assessment of total lung volumes and differentiation between overinflated, normally aerated, poorly aerated, and nonaerated lung regions. It allows for classification of different patterns of consolidation and may be predictive for the potential for recruitment. DATA SUMMARY: Experimental data suggest that HFOV at mean airway pressures (mPaw) set according to a static PV curve leads to effective lung recruitment but results in overall lung volumes that are considerably higher than those predicted from the PV relationship. In saline-lavaged sheep, similar changes in total lung volumes and subvolumes were observed during HFOV and CV. One single study specifically assessed lung volume recruitment during HFOV as compared with CV in eight patients with acute respiratory distress syndrome from pneumonia or sepsis. After 48 hrs on HFOV, total ventilated lung volume was significantly increased, whereas only a minor increase in overinflated lung volume was observed. These changes correlated with a significant improvement in gas exchange. CONCLUSION: CT is a valuable tool to quantify recruitment and overinflation during HFOV. Additional studies are needed to better characterize the specific effects of HFOV on lung volume and morphology. PMID- 15753721 TI - High-frequency oscillatory ventilation: what large-animal studies have taught us! AB - BACKGROUND: Much of the information on the physiologic effects, mechanisms of gas exchange, and potential utility of high-frequency oscillation (HFO) has been acquired in animal studies. Specifically, large animal data have been useful in assessing adult application because large animals present many of the same concerns and challenges as adults. OBJECTIVE: To review the literature on HFO testing in large animal models, identifying contributions to the understanding of mechanisms of action and the physiology of HFO. RESULTS: Large animal studies have clarified the mechanisms of gas exchange during HFO, identified approaches to setting mean airway pressure based on lung mechanics, and identified a potentially better approach to applying partial liquid ventilation. CONCLUSION: The study of HFO in large animal models has been essential to our understanding of the optimal approach to applying HFO in human studies. PMID- 15753723 TI - Noninvasive assessment of lung volume: respiratory inductance plethysmography and electrical impedance tomography. AB - OBJECTIVE: Respiratory inductance plethysmography (RIP) and electrical impedance tomography (EIT) are two monitoring techniques that have been used to assess lung volume noninvasively. METHODS: RIP uses two elastic bands around the chest and abdomen to assess global changes in lung volume. In animal models, RIP has been shown to detect changes in lung mechanics during high-frequency oscillatory ventilation and has the potential to quantify lung volumes noninvasively. EIT measures regional impedance changes with 16 electrodes around the patient's chest, each of them injecting and receiving small currents. Impedance changes have been correlated with volume changes in animal models and in humans. In a recent animal model, EIT was shown to be capable of tracking lung volume changes during high-frequency oscillatory ventilation. CONCLUSION: The promise of monitoring techniques such as RIP and EIT is that they will guide lung protective ventilation strategies and allow the clinician to optimize lung recruitment, maintain an open lung, and limit overdistension. EIT is the only bedside method that allows repeated, noninvasive measurements of regional lung volumes. In the future, it will be important to standardize the definitions of alveolar recruitment and ultimately demonstrate the superiority of EIT-guided ventilator management in providing lung protective ventilation. PMID- 15753724 TI - Clinical use of high-frequency oscillatory ventilation in adult patients with acute respiratory distress syndrome. AB - OBJECTIVE: High-frequency oscillatory ventilation (HFOV) is an emerging ventilatory strategy for adults that has been used successfully in the neonatal and pediatric population. This modality utilizes high mean airway pressures to maintain an open lung and low tidal volumes at a high frequency that allow for adequate ventilation while at the same time preventing alveolar overdistension. With the current understanding that excessive lung stretch and inadequate end expiratory ventilatory volume may be injurious to the lungs, HFOV seems to be the ideal lung-protective ventilatory mode. During the past 8 yrs, there have been increasing numbers of studies describing its use in adult patients with acute respiratory distress syndrome. This article aims to review the published studies of HFOV in adults with acute respiratory distress syndrome with regard to its safety and efficacy. DATA SOURCE: To assist us with our review, we did a search of MEDLINE (from 1966 to present) and EMBASE (1980 to present) databases to identify adult, clinical, English-language, research articles related to HFOV use. In addition, we reviewed relevant animal and mechanical ventilation studies. We did not perform a formal systematic review. DATA SYNTHESIS: The application of HFOV was mainly reported as a rescue ventilatory mode in adult patients with acute respiratory distress syndrome who were thought to have failed conventional ventilation. In these patients, HFOV has consistently been shown to improve oxygenation without obvious increases in complications measured. There was only one randomized, controlled trial comparing HFOV with conventional ventilation. This study showed that there was a nonsignificant trend toward a lower mortality rate in the HFOV group. In addition, HFOV was as effective and safe as conventional ventilation. Although there are limitations, multiple studies have shown that earlier initiation of HFOV in patients with severe acute respiratory distress syndrome may also be associated with a lower mortality. CONCLUSIONS: HFOV seems to be safe and effective for adults with severe acute respiratory distress syndrome who have failed conventional ventilation. Further research is needed to determine the ideal patients, timing, and optimal technique with which to provide HFOV. When considering HFOV as an early, lung-protective mode of ventilation, there is still a need to perform an adequately powered, randomized, controlled trial comparing it with the best available form of conventional ventilation. However, we believe that such a trial should wait until we have a better understanding of HFOV in adults. PMID- 15753725 TI - Use of high-frequency oscillatory ventilation in burn patients. AB - BACKGROUND: Patients with major burn injuries frequently develop acute respiratory distress syndrome (ARDS). High-frequency oscillatory ventilation (HFOV) has been used successfully in our regional burn center since 1999 for the management of oxygenation failure secondary to ARDS and as a method of intraoperative ventilation to allow surgical burn wound excision to proceed, despite the presence of severe ARDS. OBJECTIVE: The objective of this article is to review the use of HFOV in burn patients, with an emphasis on the indications and selection criteria for the initiation of HFOV, special considerations for patients with smoke inhalation injury, and our approach to intra-operative HFOV. SETTING: Adult regional burn center in a university-affiliated tertiary care hospital. RESULTS: We have now used HFOV in 36 severely burned patients, 33% of whom had an associated smoke inhalation injury. HFOV produced significant improvements in oxygenation among burn patients with oxygenation failure secondary to ARDS. HFOV produced a slower and less robust reversal of oxygenation failure in those with smoke inhalation compared with patients with burns alone. HFOV has been used intraoperatively for 33 procedures in 18 patients without complications. CONCLUSION: HFOV has been an indispensable ventilation modality in our burn center, and has played an important role in reversing oxygenation failure in patients with ARDS and in facilitating early excision and closure of the burn wound in these patients. PMID- 15753726 TI - High-frequency oscillatory ventilation and adjunctive therapies: inhaled nitric oxide and prone positioning. AB - OBJECTIVE: To review the use of high-frequency oscillatory ventilation (HFOV) with adjunctive therapies (inhaled nitric oxide [iNO] and prone positioning [PP]) in adult patients with acute respiratory distress syndrome (ARDS). DATA SOURCES: Published studies evaluating the use of iNO, PP, and HFOV in adult patients with ARDS. DATA SUMMARY: Despite ongoing preclinical and clinical research, the therapeutic armamentarium for ARDS remains limited. Although a pressure- and volume-limited strategy aimed at mitigating ventilator-associated lung injury has demonstrated mortality benefit, patients with severe ARDS may still develop life threatening hypoxemia. As a result, various salvage therapies aimed at improving oxygenation, including HFOV, iNO, and PP alone or in combination, have been evaluated in patients with refractory ARDS. Although the few preclinical and clinical trials of combination therapy to date have shown promising improvements in oxygenation and other physiological variables, with few adverse clinical events, the impact on survival awaits the performance of large randomized trials. CONCLUSIONS: There is limited clinical data to recommend the widespread use of combination therapy in patients with ARDS. In the subset of patients with life threatening hypoxemia from refractory ARDS, combination therapy is safe and may be considered for salvage therapy. More rigorous randomized, controlled trials are needed to help delineate the therapeutic role of combination therapy in adults with ARDS. PMID- 15753727 TI - Inhaled alternatives to nitric oxide. AB - OBJECTIVE: Inhaled nitric oxide has gained an established place in the management of pulmonary hypertension. However, cost, potential toxicity, and the lack of positive outcome data with inhaled nitric oxide therapy has generated interest in alternative inhaled, selective pulmonary vasodilators. This article describes those alternatives that have been studied to date. DESIGN: Literature review of inhaled, selective pulmonary vasodilators other than nitric oxide. METHODS: A review of the molecular mechanisms, potential side effects, and the studies to date in both animal models and clinical studies describing the physiologic effects of alternative agents to inhaled nitric oxide. CONCLUSION: There are a number of available agents that have comparable physiologic effects as inhaled nitric oxide. The best studied of these are the inhaled prostanoids (prostacyclin and iloprost), and there is growing interest in novel therapies such as phosphodiesterase inhibitors and neuropeptides. PMID- 15753728 TI - High-frequency oscillatory ventilation in adults: respiratory therapy issues. AB - OBJECTIVE: To summarize clinical information and assessment techniques relevant to respiratory therapists caring for adult patients on high-frequency oscillatory ventilation (HFOV). DATA SOURCE: Review of observational studies, controlled trials, case reports, institutional experience, and hospital HFOV guidelines for adult patients. DATA SUMMARY: Respiratory therapists require unique physical assessment skills and knowledge in managing patients on HFOV. Respiratory therapy procedures relevant to HFOV include setting endotracheal tube cuff leaks, performing lung recruiting maneuvers, endotracheal suctioning, and monitoring ventilator parameters. Respiratory therapists serve as essential team members in the creation and implementation of written HFOV guidelines (e.g., algorithms) to optimize patient care. CONCLUSION: Respiratory therapy assessment and procedural skills are essential in providing optimal care to adult patients on HFOV. PMID- 15753729 TI - Nursing and infection-control issues during high-frequency oscillatory ventilation. AB - OBJECTIVES: To review the specific nursing and infection-control issues that arise during the care of patients receiving high-frequency oscillatory ventilation (HFOV). DATA SOURCE: Published articles, governmental guidelines, and hospital procedures and practices. DATA SUMMARY: Nurses, respiratory therapists, and other clinicians caring for patients receiving HFOV need to be aware of specific differences in patient assessment, including close observation for symmetric chest-wall vibrations. In addition, management of sedation with or without neuromuscular blockade and effective communication with the patients are essential nursing skills needed with the use of HFOV. From an infection-control standpoint, HFOV is considered a high-risk respiratory procedure because of the inability to effectively filter all respiratory secretions. Appropriate infection control precautions, including patient location and use of personal protective equipment, need to be considered when implementing HFOV in the intensive care unit. CONCLUSIONS: Important infection-control and nursing issues exist that are specific to the use of HFOV. These issues should be addressed with appropriate staff education before the implementation of HFOV in an intensive care unit. PMID- 15753730 TI - Sedation, analgesia, and neuromuscular blockade for high-frequency oscillatory ventilation. AB - OBJECTIVE: To provide a comprehensive review of the issue related to the administration of sedative, analgesic, and neuromuscular blocking agents (NMBA) to patients who are receiving ventilatory support for acute respiratory distress syndrome (ARDS) with high-frequency oscillatory ventilation. RESULTS: Sedative, analgesic, and NMBA are used with great frequency in patients with severe ARDS who are undergoing high-frequency oscillatory ventilation. In particular, the use of NMBA has been higher than for other ARDS populations. Important considerations for effective treatment include careful patient evaluation, patient-based medication selection, identification of treatment goals with periodic re assessment, titration of medications to objective parameters such as sedation scales and peripheral nerve stimulation, use of intermittent therapy when feasible, implementation of drug interruption strategies, and discontinuation of medications at the earliest possible time. It is important to recognize that patients evolve from severe ARDS through phases of recovery to the resolution of respiratory failure and that ventilatory management, as well as sedative and related medication requirements, will vary markedly over the course of this process. CONCLUSIONS: A multidisciplinary, structured approach that is based on the considerations described should help achieve optimal results in this challenging patient population. PMID- 15753731 TI - Prognostic determinants of acute respiratory distress syndrome in adults: impact on clinical trial design. AB - OBJECTIVE: The objective of this study was to review known clinical predictors and biologic markers of adverse clinical outcomes in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) that might be used as selection criteria in clinical trials of novel therapies for ALI/ARDS. DATA SOURCE: Published studies on clinical predictors and biologic markers of adverse outcomes in ALI/ARDS. MAIN RESULTS: In large epidemiologic studies, a number of clinical factors have been identified consistently as independent predictors of mortality in ALI/ARDS. These include age, comorbidities, including chronic liver disease and immunosuppression, severity of illness scores, and the degree of multisystem organ failure. Several biologic markers of mortality have also been identified in large studies, including von Willebrand factor antigen, surfactant protein D, protein C, plasminogen activator inhibitor-1, interleukins 6 and 8, and the TNF receptors. The Pao2/Fio2 ratio at the onset of ALI/ARDS does not predict clinical outcome but may be more useful after the first day of ALI/ARDS. A persistently low Pao2/Fio2 ratio is associated with worse outcomes and may be a marker of failure to respond to conventional therapy. Changes in IL-6, IL-8, TNF receptors, and SP-D over the first 3 days of ALI/ARDS are also associated with adverse clinical outcomes. The use of a combination of clinical factors and biologic markers is a promising strategy that needs to be prospectively validated. CONCLUSIONS: The design of clinical trials for new therapies for ALI and ARDS is a complex problem that ultimately will have a major impact on both trial outcome and generalizability. A number of clinical factors and biologic markers can be used to differentiate groups of patients at highest risk for adverse clinical outcomes. Whether enriching study populations with these sicker patients will increase or decrease the likelihood of a treatment effect for a given therapy is unknown. PMID- 15753732 TI - Protocols for lung protective ventilation. AB - Protocols have a well-established role in clinical research and are increasingly being used to direct routine clinical care. In this article, we review the differing goals of research and clinical protocols and outline the similar process for their development. We use the mechanical ventilation protocol of the ARDS Network trial comparing small with traditional tidal volumes as an example. As a starting point for debate, we also suggest guiding principles and specific components of a protocol for high-frequency oscillatory ventilation. PMID- 15753733 TI - Other approaches to open-lung ventilation: airway pressure release ventilation. AB - OBJECTIVE: To review the use of airway pressure release ventilation (APRV) in the treatment of acute lung injury/acute respiratory distress syndrome. DATA SOURCE: Published animal studies, human studies, and review articles of APRV. DATA SUMMARY: APRV has been successfully used in neonatal, pediatric, and adult forms of respiratory failure. Experimental and clinical use of APRV has been shown to facilitate spontaneous breathing and is associated with decreased peak airway pressures and improved oxygenation/ventilation when compared with conventional ventilation. Additionally, improvements in hemodynamic parameters, splanchnic perfusion, and reduced sedation/neuromuscular blocker requirements have been reported. CONCLUSION: APRV may offer potential clinical advantages for ventilator management of acute lung injury/acute respiratory distress syndrome and may be considered as an alternative "open lung approach" to mechanical ventilation. Whether APRV reduces mortality or increases ventilator-free days compared with a conventional volume-cycled "lung protective" strategy will require future randomized, controlled trials. PMID- 15753734 TI - High-frequency percussive ventilation. AB - OBJECTIVE: To review the technique and clinical application of high-frequency percussive ventilation in critically ill patients. DESIGN: Literature search and descriptive review. RESULTS: High-frequency percussive ventilation is a time cycled, pressure-limited mode of ventilation that delivers subphysiologic tidal volumes at rates that can exceed 500 breaths/min. It offers the potential advantage over conventional ventilation of providing equal or improved oxygenation and ventilation at lower peak and end-expiratory pressures. This modality has been used to manage severe lung disease in the neonatal and pediatric population, treat inhalation injury in pediatric and adult patients, and as salvage therapy in adult patients with acute respiratory distress syndrome. CONCLUSIONS: High-frequency percussive ventilation has been shown to provide favorable gas exchange in several well-defined patient populations. It reliably improves oxygenation and provides adequate ventilation at lower peak pressures than conventional ventilation. Adequately powered, randomized, prospective studies demonstrating significant mortality benefit have not yet been performed. PMID- 15753735 TI - Combining high-frequency oscillatory ventilation and recruitment maneuvers in adults with early acute respiratory distress syndrome: the Treatment with Oscillation and an Open Lung Strategy (TOOLS) Trial pilot study. AB - OBJECTIVE: To determine the safety, feasibility, and lung-recruitment efficacy of an explicit ventilation protocol combining high-frequency oscillatory ventilation and recruitment maneuvers. DESIGN: Prospective, multiple-center, single intervention pilot study. SETTING: Four university-affiliated intensive care units. PATIENTS: Twenty-five patients with early acute respiratory distress syndrome and severe oxygenation failure. INTERVENTIONS: Patients were transitioned from standardized conventional ventilation to high-frequency oscillatory ventilation beginning with an initial cycle of up to three sustained inflation recruitment maneuvers (40 cm H2O x 40 secs), followed by a decremental titration of Fio2 and then mean airway pressure. Recruitment maneuvers were repeated for hypoxemia and routinely at least twice daily if the Fio2 was >0.4. A specific protocol was used for weaning high-frequency oscillatory ventilation, for transitioning to conventional ventilation, and for judging intolerance of conventional ventilation whereby patients should be put back on high-frequency oscillatory ventilation. MEASUREMENTS AND MAIN RESULTS: Patients (median [interquartile range] Acute Physiology and Chronic Health Evaluation II, 24 [19 32]; age, 50 [41-64]) were enrolled after 13 (range, 6-51) hrs of conventional ventilation. Following the initial cycle of recruitment, the mean (+/-sd) Pao2/Fio2 increased significantly compared with standardized conventional ventilation (200 +/- 117 vs. 92 +/- 36 mm Hg, p < .001). After a mean of 12 hrs of high-frequency oscillatory ventilation, the mean Fio2 was significantly reduced compared with prestudy levels (0.5 +/- 0.2 vs. 0.9 +/- 0.1, p < .001). A median of seven (four to 11) recruitment maneuvers was performed per patient over the study period, with only eight of 244 (3.3%) being aborted. Six of 19 patients transitioned to conventional ventilation (32%) were deemed intolerant and were switched back to high-frequency oscillatory ventilation. Protocol adherence was excellent with documented rates >90%. CONCLUSIONS: The combination of high frequency oscillatory ventilation and recruitment maneuvers resulted in rapid and sustained improvement in oxygenation, likely through lung recruitment. This explicit high-frequency oscillatory ventilation protocol appears well tolerated, feasible, and physiologically sound. PMID- 15753736 TI - Comparison between dobutamine and levosimendan for management of postresuscitation myocardial dysfunction. AB - OBJECTIVE: To investigate the effects of levosimendan, a nonadrenergic inotropic calcium sensitizer, in comparison with adrenergic dobutamine for the management of postresuscitation myocardial dysfunction following resuscitation from prolonged cardiac arrest. DESIGN: Randomized prospective animal study. SETTING: Animal research laboratory. SUBJECTS: Male Yorkshire-cross domestic pigs INTERVENTIONS: Ventricular fibrillation was induced in male domestic pigs weighing between 35 and 40 kg. Cardiopulmonary resuscitation, including precordial compression and mechanical ventilation, was started after 7 mins of untreated cardiac arrest. Electrical defibrillation was attempted after 5 mins of cardiopulmonary resuscitation. Each animal was successfully resuscitated without pharmacologic intervention. Resuscitated animals were randomized to treatment with levosimendan, dobutamine, or saline placebo. The inotropic agents or an equivalent volume of placebo diluents was administered 10 mins after restoration of spontaneous circulation. Levosimendan was administered in a loading dose of 20 microg.kg over 10 mins followed by a 220-min infusion of 0.4 microg.kg.min. Dobutamine was infused into the right atrium in an amount of 5 microg.kg.min. Treatment was continued for a total of 230 mins. MEASUREMENTS AND MAIN RESULTS: Levosimendan and dobutamine produced comparable increases in cardiac output. However, levosimendan produced significantly greater left ventricular ejection fraction and fractional area changes compared with dobutamine and saline placebo. CONCLUSIONS: Levosimendan has the potential of improving postresuscitation myocardial function. It is likely to serve as an alternative to dobutamine as an inotropic agent for management of postresuscitation myocardial dysfunction. PMID- 15753737 TI - Expression of apoptosis regulatory factors during myocardial dysfunction in endotoxemic rats. AB - OBJECTIVES: To document the time course of apoptosis pathway activation in sepsis and to determine whether Bcl-2 overexpression would improve endotoxin-induced myocardial dysfunction and mortality rate. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Sprague Dawley rats, wild-type C57BL/6 female mice, C57BL/6 female mice overexpressing Bcl-2. INTERVENTIONS: Hearts were isolated from rats treated with endotoxin (10 mg/kg, intravenously) to perform heart function, immunohistochemistry (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick 3'-end labeling, caspase 3), RNase protection assay, reverse transcriptase polymerase chain reaction, Western blotting (caspase 3), and radiolabeled annexin V studies. Twenty-four hours before endotoxin challenge (10 mg/kg, intravenously), rats were pretreated with saline or endotoxin (0.5 mg/kg, intraperitoneally), with or without parthenolide (1 mg/kg, intraperitoneally). Isolated hearts were used to test myocardial function. Mortality induced by endotoxin (10 mg/kg, intraperitoneally) was tested on wild-type or mice overexpressing Bcl-2. MEASUREMENTS AND MAIN RESULTS: Endotoxin-induced heart dysfunction was maximal at 4 and 8 hrs postinjection, started to improve, and was fully restored at 24 hrs after endotoxin treatment. Endotoxin also induced phosphatidylserine outer leaflet membrane exposure, caspase 3 activation, nuclear apoptosis, and changes in apoptosis gene expression. Bcl-2 overexpression induced by endotoxin pretreatment prevented endotoxin-induced myocardial dysfunction. Mice overexpressing Bcl-2 had dramatic improvement in survival rate compared with wild-type mice. CONCLUSIONS: These observations suggest that both death receptor and caspase-mediated apoptosis processes are activated in this sepsis model. Bcl-2 overexpression before endotoxin challenge prevents myocardial dysfunction in rats and improves survival rate in mice. PMID- 15753738 TI - Number needed to treat and cost of recombinant human erythropoietin to avoid one transfusion-related adverse event in critically ill patients. AB - OBJECTIVE: To calculate the absolute risk reduction of transfusion-related adverse events, the number of patients needed to treat, and cost to avoid one transfusion-related adverse event by using erythropoietin in critically ill patients DESIGN: Number needed to treat with sensitivity analysis. SETTING: Teaching hospital. PATIENTS: Hypothetical cohort of critically ill patients who were candidates to receive erythropoietin. INTERVENTIONS: Using vs. not using erythropoietin to reduce the need for packed red blood cell transfusions. MEASUREMENTS AND MAIN RESULTS: We used published estimates of known transfusion risks: transfusion-related acute lung injury, transfusion-related errors, hepatitis B and C, human immunodeficiency virus, human T-cell lymphotropic virus, and bacterial contamination, stratified by severity. Based on the estimated risk and frequency of transfusions with and without erythropoietin, we calculated the absolute risk reduction of transfusion-related adverse events, the number needed to treat, and cost to avoid one transfusion-related adverse event by using erythropoietin. The estimated incidence of transfusion-related adverse event was 318 permillion units transfused for all transfusion-related adverse events, 58 per million for serious transfusion-related adverse events, and 21 per million for likely fatal transfusion-related adverse events. The routine use of erythropoietin resulted in an absolute risk reduction of 191 per million for all transfusion-related adverse events, 35 per million for serious transfusion related adverse events, and 12 per million for likely fatal transfusion-related adverse events. The number needed to treat was 5,246 to avoid one transfusion related adverse event, 28,785 to avoid a serious transfusion-related adverse event, and 81,000 for a likely fatal transfusion-related adverse event. The total cost was $4,700,000 to avoid one transfusion-related adverse event, $25,600,000 to avoid one serious transfusion-related adverse event, and $71,800,000 to avoid a likely fatal transfusion-related adverse event. The magnitude of these results withstood extensive sensitivity analysis. CONCLUSIONS: From the perspective of avoidance of adverse events, erythropoietin does not appear to be an efficient use of limited resources for routine use in critically ill patients. PMID- 15753739 TI - Modulation of asymmetric dimethylarginine in critically ill patients receiving intensive insulin treatment: a possible explanation of reduced morbidity and mortality? AB - OBJECTIVE: Asymmetric dimethylarginine, which inhibits production of nitric oxide, has been shown to be a strong and independent predictor of mortality in critically ill patients with clinical evidence of organ dysfunction. Interestingly, intensive insulin therapy in critically ill patients improved morbidity and mortality, but the exact mechanisms by which these beneficial effects are brought about remain unknown. Therefore, we aimed to investigate whether modulation of asymmetric dimethylarginine concentrations by intensive insulin therapy is involved in these effects. DESIGN: A prospective, randomized, controlled trial. SETTING: A 56-bed predominantly surgical intensive care unit in a tertiary teaching hospital. PATIENTS: From a study of 1,548 critically ill patients who were randomized to receive either conventional or intensive insulin therapy, we included 79 patients who were admitted to the intensive care unit after complicated pulmonary and esophageal surgery and required prolonged (>/=7 days) intensive care. INTERVENTIONS: Determination of asymmetric dimethylarginine concentrations. MEASUREMENTS AND MAIN RESULTS: Asymmetric dimethylarginine concentrations were determined with high-performance liquid chromatography on the day of admission, on day 2, on day 7, and on the last day at the intensive care unit. Although the asymmetric dimethylarginine levels did not change between day 0 and day 2 in patients receiving intensive insulin treatment, there was a significant increase during this period in the conventionally treated patients (p = .043). Interestingly, the mean daily insulin dose was inversely associated with the asymmetric dimethylarginine concentration on the last day (r = -.23, p = .042), and the asymmetric dimethylarginine concentration on the last day at the intensive care unit was significantly lower in the intensive insulin treatment group (p = .048). Furthermore, asymmetric dimethylarginine was positively associated with duration of intensive care unit stay, duration of ventilatory support, duration of inotropic and vasopressor treatment, number of red cell transfusions, duration of antibiotic treatment, presence of critical illness polyneuropathy, mean Acute Physiology and Chronic Health Evaluation II score, and cumulative Therapeutic Intervention Scoring System-28 score. In addition, asymmetric dimethylarginine levels in patients who died were significantly higher compared with survivors, and changes in the course of asymmetric dimethylarginine plasma concentrations were predictive for adverse intensive care unit outcome. CONCLUSIONS: Modulation of asymmetric dimethylarginine concentration by insulin at least partly explains the beneficial effects found in critically ill patients receiving intensive insulin therapy. PMID- 15753741 TI - Decreased physiologic variability as a generalized response to human endotoxemia. AB - OBJECTIVE: To test the effect in normal human volunteers of transient systemic inflammation on the variability in time-series behaviors of widely divergent physiologic measures of the human inflammatory response. DESIGN: Prospective study of human volunteers who were tested on 2 consecutive days, a control day and a treatment day. Each participant served as his or her own control. SETTING: Critical care facility of a university medical center. SUBJECTS: Subjects were eight healthy human volunteers. INTERVENTIONS: Participant subjects were tested on both a baseline day with no intervention and on a treatment day when they received 4 ng/kg intravenous Escherichia coli endotoxin. MEASUREMENTS AND MAIN RESULTS: Continuous electrocardiographic recordings and serial blood sampling (performed every 5 mins) were used to create time-series of heart rate (R-R intervals), neutrophil function (phagocytosis), and plasma cortisol concentrations. For each primary measure, we recorded a significant increase in the regularity (decreased variability) of the functional measurement as assessed by the statistical entity, approximate entropy. CONCLUSIONS: Increased regularity, or decreased variability, of organ functions is a generalized response to systemic inflammation that occurs in widely divergent systems during endotoxemia. PMID- 15753742 TI - Characteristics and outcomes of cancer patients requiring mechanical ventilatory support for >24 hrs. AB - OBJECTIVES: To describe the characteristics of a large cohort of cancer patients receiving mechanical ventilation for >24 hrs and to identify clinical features predictive of in-hospital death. DESIGN: Prospective cohort study. SETTING: Ten bed oncologic medical-surgical intensive care unit. PATIENTS: A total of 463 consecutive patients were included over a 45-month period. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data were collected on the day of admission to the intensive care unit. The intensive care unit and hospital mortality rates were 50% and 64%, respectively. There were 359 (78%) patients with solid tumors and 104 (22%) with hematologic malignancies; 35 (8%) patients had leukopenia. Sepsis (63%), coma (15%), invasion or compression by tumor (11%), pulmonary embolism (7%), and cardiopulmonary arrest (6%) were the main reasons for mechanical ventilation. The independent unfavorable risk factors for mortality were older age (odds ratio, 3.09; 95% confidence interval, 1.61-5.93, for patients 40-70 yrs old, and odds ratio, 9.26; 95% confidence interval, 4.16-20.58, for patients >70 yrs old); performance status 3-4 (odds ratio, 2.51; 95% confidence interval, 1.40 4.51); cancer recurrence/progression (odds ratio, 3.43; 95% confidence interval, 1.81-6.53); Pao2/Fio2 ratio <150 (odds ratio, 2.64; 95% confidence interval, 1.40 4.99); Sequential Organ Failure Assessment score (excluding respiratory domain, each 4 points; odds ratio, 2.34; 95% confidence interval, 1.70-3.24); and airway/pulmonary invasion or compression by tumor as a reason for mechanical ventilation (odds ratio, 5.73; 95% confidence interval, 1.92-17.08). CONCLUSIONS: Severity of acute organ failures, poor performance status, cancer status, and older age were the main determinants of mortality. The appropriate use of such easily available clinical characteristics may avoid forgoing intensive care for patients with a chance of survival. PMID- 15753743 TI - Intrinsic positive end-expiratory pressure in Acute Respiratory Distress Syndrome (ARDS) Network subjects. AB - OBJECTIVE: We tested the hypothesis that subjects randomized to the 6 mL/kg predicted body weight tidal volume study group of the National Institutes of Health Acute Respiratory Distress Syndrome (ARDS) Network study had higher levels of intrinsic positive end-expiratory pressure (PEEP) than subjects randomized to the 12 mL/kg predicted body weight tidal volume study group. DESIGN: Secondary analysis of a subgroup from a randomized controlled trial. SETTING: Hospitals located in San Francisco, CA, and Seattle, WA. PATIENTS: Eighty-four patients enrolled in the ARDS Network tidal volume trial in San Francisco, CA, and Seattle, WA, with records of measurement of intrinsic PEEP. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Intrinsic PEEP was assessed a median of six times over the first 48 hrs of ARDS Network protocol ventilation in study subjects, with no significant difference in number of measurements between subjects randomized to the tidal volume protocol of 6 mL/kg compared with 12 mL/kg. We found that intrinsic PEEP was higher among subjects randomized to the 6 mL/kg protocol, with a median intrinsic PEEP of 1.3 cm H2O (interquartile range, 0-3.1 cm H2O), compared with a median intrinsic PEEP of 0.5 cm H2O (interquartile range, 0-1.5 cm H2O) among subjects randomized to 12 mL/kg (p = .029). There was no difference in total PEEP between the study groups. CONCLUSIONS: In a subgroup of ARDS Network subjects, intrinsic PEEP was statistically significantly higher in subjects randomized to the 6 mL/kg protocol than those in the 12 mL/kg study group. The amount of intrinsic PEEP was very low in both study groups, and difference of median intrinsic PEEP between the groups was <1 cm H2O. It is unlikely that the difference in intrinsic PEEP between the study groups was clinically important in the ARDS Network study of low tidal volume ventilation. PMID- 15753744 TI - A controlled trial of smart infusion pumps to improve medication safety in critically ill patients. AB - OBJECTIVE: Intravenous medications are vital during inpatient management. Errors associated with the administration of medications through intravenous infusion pumps to critically ill patients can result in adverse drug events. We sought to assess the impact of smart pumps with integrated decision support software on the incidence and nature of medication errors and adverse drug events. DESIGN: We performed a prospective, randomized time-series trial and compared the serious medication error rate between intervention (decision support on) and control (decision support off) periods. Serious medication errors included both near misses and preventable adverse drug events. Pump software produced log reports to help identify potential events. Events were presented to physicians for rating of event type, preventability, and severity. SETTING: Cardiac surgical intensive care and step-down units between February and December 2002. PATIENTS: Pump data were available for 744 cardiac surgery admissions. INTERVENTIONS: Decision support during medication administration provided feedback including alerts, reminders, and unit-specific drug rate limits. MEASUREMENTS AND MAIN RESULTS: We found a total of 180 serious medication errors, including 14 and 11 preventable adverse drug events and 73 and 82 nonintercepted potential adverse drug events in the control and intervention periods, respectively. The serious medication error rates in the control and intervention periods were 2.03 and 2.41 per 100 patient pump-days, respectively (p = .124). We also found numerous opportunities for safety improvement. Violations of infusion practice during the intervention periods included 571 (25%) bypasses of the drug library. Medications were also frequently administered without documentation of physician orders in both periods (n = 823; 7.7%). CONCLUSION: Intravenous medication errors and adverse drug events were frequent and could be detected using smart pumps. We found no measurable impact on the serious medication error rate, likely in part due to poor compliance. Although smart pumps have great promise, technological and nursing behavioral factors must be addressed if these pumps are to achieve their potential for improving medication safety. PMID- 15753745 TI - Cytochrome P-450 activity is differentially altered in severely injured patients. AB - OBJECTIVE: To assess whether changes in cytochrome P-450 (CYP) activity of specific CYP enzymes occur in severely injured patients and to assess changes in CYP activity during recovery. DESIGN: Prospective clinical study. SETTING: University-affiliated, level I trauma center and trauma critical care unit. PATIENTS: Twenty-three multiply injured patients admitted to a trauma critical care unit were compared with healthy volunteers. INTERVENTIONS: CYP metabolizing activity was assessed using the probe drugs mephenytoin (CYP-2C19), chlorzoxazone (CYP-2E1), dapsone (multiple CYP enzymes), and flurbiprofen (CYP-2C9). MEASUREMENTS AND MAIN RESULTS: Mephenytoin metabolism was profoundly suppressed after injury and increased during postinjury recovery, whereas chlorzoxazone metabolism was suppressed to a lesser degree. Measures of dapsone and flurbiprofen metabolism were elevated throughout the study. Chlorzoxazone and mephenytoin metabolism correlated with the multiple organ dysfunction score and with the multiple organ failure score. CONCLUSIONS: CYP isoform activity is differentially altered by shock and trauma in injured patients. The metabolic activity of selected CYP isoforms may have potential for evaluating acute hepatic dysfunction in critically ill trauma patients. PMID- 15753746 TI - Five-year survival, quality of life, and individual costs of 303 consecutive medical intensive care patients--a cost-utility analysis. AB - OBJECTIVE: To assess long-term survival, health-related quality of life, and associated costs 5 yrs after discharge from a medical intensive care unit. DESIGN: Prospective cohort study. SETTING: Medical intensive care unit of a German university hospital. PATIENTS: Three hundred and three consecutive patients with predominantly cardiovascular and pulmonary disorders admitted between November 1997 and February 1998 with an intensive care unit length of stay >24 hrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Demographic data, Simplified Acute Physiology Score II, Sequential Organ Failure Assessment, simplified Therapeutic Intervention Scoring System, and individual intensive care unit and hospital costs were prospectively recorded. Primary outcomes included 5 yr survival, functional status, health-related quality of life (Medical Outcome Short Form, SF-36), effective costs per survivor, and costs per life year and per quality-adjusted life year gained. Of 303 patients, 44 (14.5%) died in the hospital. Among the remaining 259 patients, 190 (73%) survived the 5-yr follow up and 173 patients (91%) completed the questionnaire. Baseline demographics including gender, age, Simplified Acute Physiology Score II, Sequential Organ Failure Assessment, simplified Therapeutic Intervention Scoring System, and admission diagnosis were similar between hospital and long-term survivors (p > .05 for all). The health status index of those patients surviving the 5-yr follow up was 0.88, independent of patients' severity of illness. The average effective costs per survivor were 8.827 for intensive care unit costs and 14.130 for intensive care unit and hospital costs. Mean costs per life year and per quality adjusted life year gained amounted to 19.330 and 21.922 , respectively. Increasing severity of illness was associated with higher costs. CONCLUSIONS: Considering the severity of illness and the patients' outcome, the costs associated with both life year and quality-adjusted life year gained were within generally accepted limits for other potentially life-saving treatments. PMID- 15753747 TI - Effect of hypertonic saline dextran on acid-base balance in patients undergoing surgery of abdominal aortic aneurysm. AB - OBJECTIVE: To evaluate the magnitude and cause of metabolic acidosis after infusion of 7.5% sodium chloride 6% dextran 70. DESIGN: Randomized, prospective clinical study. SETTING: University hospital. PATIENTS: Two groups of 14 patients each, undergoing repair of abdominal aortic aneurysm. INTERVENTIONS: Patients were randomly assigned to receive either 250 mL of hypertonic saline dextran (HSD) or a conventional fluid regimen with 250 mL of hydroxyethyl starch in normal saline solution (H-NS) during the period of aortic clamping. Additionally, normal saline was used in both groups to reach a target pulmonary artery occlusion pressure of 15-18 mmHg. pH, Paco2, and serum concentrations of sodium, potassium, magnesium, calcium, chloride, lactate, albumin, and phosphate were measured. Strong ion difference was calculated as (sodium + potassium + magnesium + calcium) - (chloride + lactate). The amount of weak plasma acid was calculated. MEASUREMENTS AND MAIN RESULTS: The infusion of HSD resulted in an immediate large increase in serum sodium (19 mmol/L) and chloride (22 mmol/L), whereas the infusion of H-NS led only to mild increases in serum sodium (3 mmol/L) and chloride (6 mmol/L). Both HSD and H-NS caused concomitant and equal decreases in the amount of weak plasma acid, strong ion difference, and pH (7.28-7.30). The reduction of bicarbonate was also identical and proportional to the extent of dilution due to infusion of HSD and H-NS. This induced metabolic acidosis was corrected spontaneously in both groups 24 hrs after surgery. CONCLUSION: Both the intravenous administration of 7.5% sodium chloride and the conventional fluid regimen with saline-based 6% hydroxyethyl starch solution resulted in a metabolic acidosis of equal extent. This suggests dilution of plasma buffers or a decrease in strong ion difference to be the primary cause of metabolic acidosis. PMID- 15753749 TI - Projected incidence of mechanical ventilation in Ontario to 2026: Preparing for the aging baby boomers. AB - OBJECTIVE: The aging baby boomers are expected to have a significant impact on the healthcare system. Mechanical ventilation is an age-dependent, costly, and relatively nondiscretionary medical service that may be particularly affected by the aging population. We forecast the future incidence of mechanical ventilation to the year 2026 to understand the impact of aging baby boomers on critical care resources. DESIGN: Population-based, sex-specific, and age-specific mechanical ventilation incidences for adults for the year 2000 were directly standardized to population projections to estimate the incidence of mechanical ventilation, in 5 yr intervals, from 2006 to 2026. Sensitivity analyses were performed by varying population projections and mechanical ventilation incidence for the elderly. SETTING: Province of Ontario, Canada. PATIENTS: Noncardiac surgery, mechanically ventilated adults. INTERVENTIONS: None. MAIN RESULTS: The projected number of ventilated patients in 2026 was 34,478, representing an 80% increase from 2000. The crude incidence increased 31%, from 222 to 291 per 100,000 adults. The annually compounded projected growth rate during this 26-yr period was 2.3%, similar to the actual growth rate experienced in the 1990s. The projected incidence was relatively insensitive to changes in assumptions, with estimates for 2026 ranging from 31,473 to 36,313 ventilated adults. CONCLUSIONS: The incidence of mechanical ventilation projected to the year 2026 will steadily increase and outpace population growth as occurred in the 1990s. In the current environment in which intensive care unit resources are limited and ventilated patients already use a significant proportion of acute care resources, planning for this continued growth is necessary. Existing evidence-based strategies that improve both the efficiency and efficacy of critical care services should be carefully evaluated for widespread implementation. PMID- 15753748 TI - Persistent elevation of high mobility group box-1 protein (HMGB1) in patients with severe sepsis and septic shock. AB - OBJECTIVE: To study the systemic release and kinetics of high mobility group box 1 protein (HMGB1) in relation to clinical features in a population of patients with severe sepsis or septic shock and to compare these with the kinetics of the cytokines interleukin-6, interleukin-8, interleukin-10, and tumor necrosis factor alpha. DESIGN: Prospective study of two cohorts of patients. SETTING: Intensive care unit and infectious disease clinic at Karolinska University Hospital Huddinge. PATIENTS: Twenty-six patients with severe sepsis, 33 patients with septic shock, and a reference group of five patients with sepsis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Sixty-four patients were included, ten of whom died within 28 days. Cytokine levels were measured at five time points during the first week after admission and were correlated to Acute Physiology and Chronic Health Evaluation II and Sepsis-related Organ Failure Assessment scores. Two HMGB1 assays were used. Both demonstrated delayed kinetics for HMGB1 with high levels on inclusion that remained high throughout the study period. Serum concentration at 144 hrs, the last sampling point, was 300 times higher, 34,000 +/- 76,000 pg/mL (mean +/- sd), than any of the other cytokines. This study, however, found no predictable correlation between serum levels of HMGB1 and severity of infection. We did quite unexpectedly find significantly lower levels of HMGB1 in nonsurvivors compared with survivors as measured by our main assay, but the other showed no difference between the two groups. Levels of interleukin 6, interleukin-8, interleukin-10, and tumor necrosis factor-alpha correlated significantly with severity of disease, and all were significantly higher in patients with septic shock compared with those with severe sepsis. Neither of these comparisons showed significant correlations for HMGB1. CONCLUSIONS: This is the first prospective study assessing the release over time of HMGB1 in a population of patients with sepsis, severe sepsis, or septic shock. Levels remained high in the majority of patients up to 1 wk after admittance, indicating that the cytokine indeed is a downstream and late mediator of inflammation. Further studies are required to fully define the relationship of HMGB1 to severity of disease. PMID- 15753750 TI - Comparison of two bispectral index algorithms in monitoring sedation in postoperative intensive care patients. AB - OBJECTIVE: Comparison of two versions of bispectral index (BIS) derived from the electroencephalogram in mechanically ventilated and continuously sedated patients after major abdominal surgery. DESIGN: Prospective, cohort, observational, unblinded study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: Forty-six patients undergoing major abdominal surgery scheduled for postoperative mechanical ventilation and continuous sedation. INTERVENTIONS: Patients were continuously sedated using propofol and sufentanil. MEASUREMENTS AND MAIN RESULTS: Electrodes for determination of BIS were placed at the forehead of the patients according to the manufacturer's specifications immediately after arrival to the intensive care unit. The level of consciousness was assessed every 2 hrs by a clinical sedation scale (Ramsay sedation scale, levels 1-6). BIS, electromyographic activity, and signal quality index were recorded simultaneously at the same time as the Ramsay scale for 24 hrs or until patients were ready for extubation. BIS (version 2.10) and BIS XP (version 3.12) were determined at 2-hr intervals. BIS and BIS XP showed a high correlation of readings (rs = .79, p < .01). However, the methods did not yield identical results. Both variables were significantly influenced by electromyographic activity, especially at high values, whereas there was only a weak correlation with body temperature. Some of the deeply sedated patients (Ramsay 5 or 6) had BIS readings >80 (BIS, 16%; BIS XP, 13%; p = not significant). CONCLUSIONS: The newer algorithm BIS XP did not perform better than the previous version BIS in patients after major surgery who were mechanically ventilated and sedated on an intensive care unit. This precludes the use of BIS or BIS XP for distinguishing, among deeply sedated ICU patients, those with and without preserved cerebral electrical activity. PMID- 15753751 TI - Ambient isoflurane pollution and isoflurane consumption during intensive care unit sedation with the Anesthetic Conserving Device. AB - OBJECTIVE: To examine ambient isoflurane pollution, scavenging efficacy, and isoflurane consumption using the Anesthetic Conserving Device (ACD) for prolonged isoflurane sedation in the intensive care unit. DESIGN: Prospective observational study. SETTING: Multidisciplinary university intensive care unit. PATIENTS: Fifteen adult ventilator-dependent intensive care unit patients sedated with isoflurane for 12-96 hrs. INTERVENTIONS: Isoflurane was infused to the ACD for sedation of study subjects. Changing of the ACD, isoflurane syringe, and opening of the respiratory circuit were performed in a standardized fashion according to investigator instructions. Active scavenging of waste gas from the ventilator was performed in ten patients; in five patients no active scavenging was performed. MEASUREMENTS AND MAIN RESULTS: Continuous spectrophotometric measurement of ambient isoflurane pollution in parts per million (ppm) at 0.5 m from the patient's head and passive lapel dosimeter sampling for ten staff nurses over 8 hr shifts. Isoflurane requirement and agent consumption were registered in all patients. Spectrophotometric readings (0.1 +/- 0.2 ppm) were well below internationally recommended long-term exposure limits in all cases. Isoflurane peaks during nursing procedures were brief, infrequent, and of low amplitude. There was no observed difference between isoflurane trace levels with or without an active scavenging system in use. Passive dosimeter values were also low, ranging from undetectable to 0.16 ppm. Mean isoflurane consumption was 2.1 +/- 1.0 mL/hr. This is approximately one fourth of predicted and previously reported consumption of isoflurane with vaporizer-administered sedation in the intensive care unit setting. CONCLUSIONS: In the present setting, isoflurane via the ACD is an environmentally safe method of sedation provided users follow instructions for standardizing procedures with potential spillage of isoflurane. This method of sedation requires considerably less isoflurane than with traditional vaporizer technique. PMID- 15753752 TI - Cardiovascular response to acute normovolemic hemodilution in patients with coronary artery diseases: Assessment with transesophageal echocardiography. AB - OBJECTIVE: Preoperative acute normovolemic hemodilution induces an increase in circulatory output that is thought to be limited in patients with cardiac diseases. Using multiple-plane transesophageal echocardiography, we investigated the mechanisms of cardiovascular adaptation during acute normovolemic hemodilution in patients with severe coronary artery disease. DESIGN: Prospective case-control study. SETTING: Operating theater in a university hospital. PATIENTS: Consecutive patients treated with beta-blockers, scheduled to undergo coronary artery bypass (n = 50). INTERVENTIONS: After anesthesia induction, blood withdrawal and isovolemic exchange with iso-oncotic starch (1:1.15 ratio) to achieve a hematocrit value of 28%. MEASUREMENTS AND MAIN RESULTS: In addition to heart rate and intravascular pressures, echocardiographic recordings were obtained before and after acute normovolemic hemodilution to assess cardiac preload, afterload, and contractility. In a control group, not subjected to acute normovolemic hemodilution, hemodynamic variables remained stable during a 20-min anesthesia period. Following acute normovolemic hemodilution, increases in cardiac stroke volume (+28 +/- 4%; mean +/- sd) were correlated with increases in central venous pressure (+2.0 +/- 1.3 mm Hg; R = .56) and in left ventricular end diastolic area (+18 +/- 5%, R = .39). The unchanged left ventricular end-systolic wall stress and preload-adjusted maximal power indicated that neither left ventricular afterload nor contractility was affected by acute normovolemic hemodilution. Diastolic left ventricular filling abnormalities (15 of 22 cases) improved in 11 patients and were stable in the remaining four patients. Despite reduction in systemic oxygen delivery (-20.5 +/- 7%, p < .05), there was no evidence for myocardial ischemia (electrocardiogram, left ventricular wall motion abnormalities). CONCLUSIONS: In anesthetized patients with coronary artery disease, moderate acute normovolemic hemodilution did not compromise left ventricular systolic and diastolic function. Lowering blood viscosity resulted in increased stroke volume that was mainly related to increased venous return and higher cardiac preload. PMID- 15753753 TI - Increased leakage of sarcoplasmic reticulum Ca2+ contributes to abnormal myocyte Ca2+ handling and shortening in sepsis. AB - OBJECTIVE: Changes in cardiac function due to sepsis have been widely reported. However, the underlying mechanisms remain poorly understood. In the mammalian heart, myocyte function and intracellular calcium homeostasis are closely coupled. In this study we tested the hypothesis that alterations in cardiac calcium homeostasis due to sepsis underlie the observed myocyte dysfunction. DESIGN: Randomized prospective animal study. SETTING: Research laboratory. SUBJECTS: Male Sprague-Dawley rats weighing 250-275 g. INTERVENTIONS: We induced sepsis by cecal ligation and puncture in the rat, which mimics the type of infection caused by perforation of the intestine in humans. MEASUREMENTS AND RESULTS: Forty-eight hours after cecal ligation and puncture, isolated cardiac ventricular cardiomyocytes demonstrated a 57% decreased peak systolic [Ca]. The time constant of the Ca transient increased 71% and 57% in myocytes obtained 24 hrs and 48 hrs after cecal ligation and puncture, respectively. The average shortening of cardiomyocytes 48 hrs after cecal ligation and puncture was significantly decreased. To investigate the cellular mechanisms of altered Ca transients and myocyte shortening, we measured Ca sparks, the spontaneous local Ca release events in cardiomyocytes at resting states. The Ca spark frequency progressively increased in myocytes 24 hrs and 48 hrs after cecal ligation and puncture. The total activity of sparks also increased compared with sham-operated animals. The overall leakage of sarcoplasmic reticulum Ca in resting states was increased in sepsis and resulted in reduced sarcoplasmic reticulum Ca content. CONCLUSIONS: Abnormal Ca leakage from the sarcoplasmic reticulum contributes significantly to the depressed myocyte shortening in sepsis. In the future, modalities that prevent this Ca leakage may prove beneficial in the treatment of sepsis-induced myocyte shortening. PMID- 15753754 TI - Sodium/hydrogen exchange activity in sepsis and in sepsis complicated by previous injury: 31P and 23Na NMR study. AB - OBJECTIVE: Sepsis or septic shock occurs frequently in sick and injured patients and is associated with a significant mortality. Myocardial contractile dysfunction has been proposed to be a major determinant of sepsis-related mortality. This study was directed to examine the role of Na/H exchange activity in myocardial defects after sepsis or after sepsis complicated by a previous burn injury. DESIGN: Laboratory study. SETTING: University research laboratory. SUBJECTS: Sprague-Dawley rats (300-350 g, males). INTERVENTIONS: Cardiac function, cellular Na and Ca, myocardial pH, and high-energy phosphates were examined in perfused hearts harvested after sepsis alone (intratracheal Streptococcus pneumoniae, 0.4 mL of 1 x 10 CFU/mL), after sepsis complicated by previous burn injury (40% total body surface area), and after amiloride (a selective inhibitor of Na/H exchange) treatment of either sepsis alone or sepsis plus burn. MEASUREMENTS AND RESULTS: The ratio of Na signal from the intracellular compartment (Nai) compared with an external standard (monitored by Na-NMR spectroscopy, TmDOTP shift reagent) increased by 70% in sepsis alone and by 41% in sepsis complicated by previous burn injury compared with shams. Cardiac adenosine triphosphate and intracellular pH (P nuclear magnetic resonance spectroscopy) were unchanged by sepsis or sepsis plus burn. Left ventricular pressure and maximal change in pressure over time were reduced after sepsis or after sepsis plus burn injury. Amiloride treatment in either sepsis or sepsis complicated by a previous burn injury prevented myocardial Na and Ca accumulation, attenuated sepsis-related lactic acidosis, and improved left ventricular function. CONCLUSION: Our results suggest that sepsis-related cardiac dysfunction is mediated, in part, by Na/H exchange activity, and inhibition of Na/H exchange activity improves cardiac function after sepsis alone or sepsis complicated by a previous injury. PMID- 15753755 TI - Nebulized nitric oxide/nucleophile adduct reduces pulmonary vascular resistance in mechanically ventilated septicemic sheep. AB - OBJECTIVE: To study the effects of a novel, intermittently administered, aerosolized nitric oxide donor, methyl-N-2-dimethylaminoethyl-3 aminoproprionid/nitric oxide (DMDE-NO), on pulmonary hemodynamic responses to sepsis. DESIGN: Prospective, randomized, controlled study in awake sheep. SETTING: Investigational intensive care unit of a university medical center. SUBJECTS: Thirteen instrumented merino ewes weighing 36 +/- 0.9 kg underwent a hemodynamic study 1 wk postoperatively. INTERVENTIONS: On the day of the experiment, the sheep received a tracheotomy and mechanical ventilation was subsequently started. Pseudomonas aeruginosa bacteria were infused intravenously, beginning at time 0 hrs and continuing throughout the 48-hr experiment. The animals were randomly assigned to receive nebulized DMDE-NO 1 mg/kg, dissolved in 8 mL of saline (DMDE-NO group, n = 7), or nebulized saline alone (control group, n = 6) delivered by a nebulizer. The nebulizations started at 2, 6, 20, 24, and 43 hrs after the baseline, each time lasting for 1 hr. MEASUREMENTS AND MAIN RESULTS: Inhaled aerosolized DMDE-NO reversibly reduced the sepsis-induced increase in pulmonary artery pressure by 13-17% and pulmonary vascular resistance index by 21-31% compared with the values registered before the administration of the drug. Systemic hemodynamics underwent an early hypodynamic phase followed by a gradual increase in cardiac index and a decrease in both mean arterial pressure and systemic vascular resistance index, but with no significant difference between groups. Gas exchange variables and plasma nitrite/nitrate did not differ significantly between groups either. CONCLUSIONS: In sheep, inhaled nebulized DMDE-NO reduces sepsis-induced changes in pulmonary hemodynamics with no change in systemic hemodynamics or gas exchange. PMID- 15753756 TI - Calpain inhibition decreases endothelin-1 levels and pulmonary hypertension after cardiopulmonary bypass with deep hypothermic circulatory arrest. AB - OBJECTIVE: Cardiopulmonary bypass in infants and children can result in cardiopulmonary dysfunction through ischemia and reperfusion injury. Pulmonary hypertension and injury are particularly common and morbid complications of neonatal cardiac surgery. Inhibition of calpain, a cysteine protease, has been shown to inhibit reperfusion injury in adult organ systems. The hypothesis is that calpain inhibition can alleviate the cardiopulmonary dysfunction seen in immature animals following ischemia and reperfusion with cardiopulmonary bypass. DESIGN: Animal case study. SETTING: Medical laboratory. SUBJECTS: Crossbred piglets (5-7 kg). INTERVENTIONS: Piglets were cooled with cardiopulmonary bypass to 18 degrees C followed by deep hypothermic circulatory arrest for 120 mins. Animals were rewarmed to 38 degrees C on cardiopulmonary bypass and maintained for 120 mins. Six animals were administered calpain inhibitor (Z-Leu-Leu-Tyr fluoromethyl ketone; 1 mg/kg, intravenously) 60 mins before cardiopulmonary bypass. Nine animals were administered saline as a control. Plasma endothelin-1, pulmonary and hemodynamic function, and markers of leukocyte activity and injury were measured. MEASUREMENTS AND MAIN RESULTS: Calpain inhibition prevented the increased pulmonary vascular resistance seen in control animals (95.7 +/- 39.4 vs. 325.3 +/- 83.6 dyne.sec/cm, respectively, 120 mins after cardiopulmonary bypass and deep hypothermic circulatory arrest, p = .05). The attenuation in pulmonary vascular resistance was associated with a blunted plasma endothelin-1 response (4.91 +/- 1.72 pg/mL with calpain inhibition vs. 10.66 +/- 6.21 pg/mL in controls, p < .05). Pulmonary function after cardiopulmonary bypass was better maintained after calpain inhibition compared with controls: Po2/Fio2 ratio (507.2 +/- 46.5 vs. 344.7 +/- 140.5, respectively, p < .05) and alveolar-arterial gradient (40.0 +/- 17.2 vs. 128.1 +/- 85.2 mm Hg, respectively, p < .05). Systemic oxygen delivery was higher after calpain inhibition compared with controls (759 +/- 171 vs. 277 +/- 46 mL/min, respectively, p < .001). In addition, endothelial nitric oxide synthase activity in lung tissue was maintained with calpain inhibition. CONCLUSIONS: The reduction in plasma endothelin-1 and maintenance of lung endothelial nitric oxide levels after cardiopulmonary bypass and deep hypothermic circulatory arrest with calpain inhibition were associated with reduced pulmonary vascular resistance. Improved gas exchange and higher systemic oxygen delivery suggest that calpain inhibition may be advantageous for reducing postoperative cardiopulmonary dysfunction commonly associated with pediatric heart surgery and cardiopulmonary bypass. PMID- 15753757 TI - Influence of heme-based solutions on stress protein expression and organ failure after hemorrhagic shock. AB - OBJECTIVE: Hemoglobin-based oxygen carriers (e.g., diaspirin-cross-linked hemoglobin [DCLHb] and hemoglobin glutamer-200 [HbG]) may have potential in the treatment of hemorrhagic shock. The nitric oxide scavenging and direct vasoconstrictive side effects of free hemoglobin of currently available preparations may increase organ injury after shock in contrast to non-oxygen carrying heme solutions (e.g., hemin arginate [HAR]). However, both classes of substances might induce the protective enzyme heme oxygenase (HO)-1, particularly in the liver. The aim of the study was to assess the role of pretreatment with DCLHb, HbG, or HAR on HO-1 expression and organ injury after hemorrhagic shock. DESIGN: Prospective controlled laboratory study. SETTING: Animal research laboratory at a university hospital. SUBJECTS: Male Sprague-Dawley rats (200-300 g body weight, n = 5-12/group). INTERVENTIONS: Twenty-four hours after different doses of DCLHb, HbG (each 1, 2, or 3 g/kg of body weight), or HAR (5, 25, or 75 mg/kg of body weight), the protein expression of HO-1 and heat shock protein-70 in liver, kidney, heart, lungs, and aorta was determined. Twenty-four hours after pretreatment with DCLHb, HbG, or HAR, rats were subjected to hemorrhage (mean arterial blood pressure, 35-40 mm Hg for 1 or 2 hrs)/resuscitation (5 or 4 hrs, respectively). Animals treated with Ringer's solution (30 mL/kg of body weight) served as controls. In additional experiments, HO activity was blocked with tin mesoporphyrin-IX. MEASUREMENTS AND MAIN RESULTS: DCLHb, HbG, and HAR dose dependently induced HO-1 protein but not heat shock protein-70. Pretreatment with DCLHb or HbG shortened the onset of decompensation in shock (DCLHb, 40 +/- 11 mins; HbG, 36 +/- 4 mins) compared with vehicle (68 +/- 4 mins, p < .05) and HAR pretreatment (81 +/- 7 mins, p < .05). High doses of DCLHb pretreatment increased mortality (2 hrs of shock, 80%; p < .05 vs. vehicle or HAR). Pretreatment with HAR led to higher shed blood volumes (p < .05) and higher hepatocellular ATP levels (2 hrs of shock, p < .05 vs. DCLHb and HbG). Blockade of HO activity by tin mesoporphyrin-IX abolished the protection mediated by HAR. CONCLUSIONS: Although DCLHb, HbG, and HAR induce HO-1 in the absence of an unspecific stress response, only HAR pretreatment protects against shock-induced organ failure. Although the underlying mechanisms of positive HAR priming are not completely understood, the induction of HO-1 expression and the lack of nitric oxide scavenging through HAR may play an important role. PMID- 15753759 TI - Hepatosplanchnic blood flow control and oxygen extraction are modified by the underlying mechanism of impaired perfusion. AB - OBJECTIVE: To assess the effects of low hepatosplanchnic blood flow on regional blood flow control and oxygenation. DESIGN: Three randomized, controlled animal experiments. SETTING: Two university experimental research laboratories. SUBJECTS: Pigs of either gender. INTERVENTIONS: Isolated abdominal blood flow reduction: An extracorporeal shunt with reservoir and roller pump was inserted between proximal and distal aorta in 11 pigs. Abdominal aortic blood flow was reduced by 50% by activating the shunt. Mesenteric ischemia: In seven pigs, superior mesenteric arterial flow was reduced to 4 mL.kg.min for 4 hrs. Cardiac tamponade: In 12 pigs, aortic blood flow was reduced by cardiac tamponade to 50 mL (moderate tamponade) and further to 30 mL.kg.min (severe tamponade) for 1 hr each. In each experimental condition, the same number of control animals was used. MEASUREMENTS AND MAIN RESULTS: Abdominal blood flow reduction, acute mesenteric ischemia, and moderate tamponade resulted in a portal venous flow (QPV) reduction to 51 +/- 23%, 52 +/- 18%, and 61 +/- 25% (mean +/- sd) of baseline flow, respectively. During abdominal blood flow reduction, QPV and hepatic arterial flow (QHA) decreased proportionally, whereas in moderate tamponade and acute mesenteric ischemia QPV reduction was associated with an increase in QHA of 30 +/- 39% and 102 +/- 108%, respectively (p = .001 and .018). Prolonged mesenteric ischemia restored total hepatic blood flow (Qliver) completely. During all conditions, decreasing mesenteric oxygen consumption was partly prevented by increased mesenteric oxygen extraction (p < .001 for all conditions). In contrast, decreasing hepatic oxygen delivery was associated with increased oxygen extraction in tamponade (p = .009) but not in abdominal blood flow reduction. CONCLUSIONS: Blood flow redistribution can restore Qliver totally when mesenteric blood flow is reduced selectively, partially when cardiac output is reduced, and not at all during abdominal blood flow reduction. Since hepatic oxygen extraction does not increase in abdominal blood flow reduction, hepatic oxygenation is at risk in this condition. PMID- 15753758 TI - Polymorphisms in CD14, mannose-binding lectin, and Toll-like receptor-2 are associated with increased prevalence of infection in critically ill adults. AB - OBJECTIVE: To test for the association of single nucleotide polymorphisms of the innate immunity receptors cluster of differentiation (CD)-14, mannose-binding lectin, and Toll-like receptor-2 with clinical phenotype in critically ill patients with systemic inflammatory response syndrome. DESIGN: Genetic association study. SETTING: Tertiary care mixed medical-surgery intensive care unit at St. Paul's Hospital, Vancouver, BC, a teaching hospital associated with the University of British Columbia. PATIENTS: A cohort of 252 critically ill Caucasians with systemic inflammatory response syndrome. INTERVENTIONS: DNA was extracted from discarded blood. Clinical data were gathered by retrospective chart review. MEASUREMENTS AND MAIN RESULTS: C-159T CD14, the X/Y and B, C, and D polymorphisms of mannose-binding lectin, and T-16933A Toll-like receptor-2 were genotyped using polymerase chain reaction-restriction fragment length polymorphism. We tested for association of genotype with prevalence of positive bacterial cultures, type of organism (Gram-positive, Gram-negative, other), sepsis and septic shock at admission to the intensive care unit, and 28-day survival. CD14 -159TT was associated with increased prevalence of positive bacterial cultures and with Gram-negative bacteria. Mannose-binding lectin haplotype pairs XO/O and O/O were also associated with increased prevalence of positive bacterial cultures but not with a specific organism class. Toll-like receptor-2 -16933AA was associated with increased prevalence of sepsis and with Gram-positive bacteria. In contrast, the polymorphisms were not associated with increased prevalence of septic shock or altered 28-day survival. CONCLUSIONS: Single nucleotide polymorphisms in CD14, mannose-binding lectin, and Toll-like receptor-2 are associated with increased prevalence of positive bacterial cultures and sepsis but not with altered prevalence of septic shock or decreased 28-day survival. Furthermore, CD14 single nucleotide polymorphisms were associated with Gram-negative bacteria and Toll-like receptor-2 with Gram positive bacteria, whereas mannose-binding lectin was not associated with a particular organism class. Thus, single nucleotide polymorphisms in innate immunity receptors may alter recognition and clearance of bacteria without changing outcomes of critically ill adults with systemic inflammatory response syndrome. PMID- 15753760 TI - Non-neurologic organ dysfunction in severe traumatic brain injury. AB - OBJECTIVE: To describe the incidence of non-neurologic organ dysfunction and its association with outcome in patients with severe traumatic brain injury admitted to intensive care. DESIGN: Observational cohort study. SETTING: Foothills Medical Centre, which is the only neurosurgical service in southern Alberta (population approximately 1.3 million). PATIENTS: Patients were 209 consecutive patients with severe traumatic brain injury. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Non-neurologic organ dysfunction was measured by the maximum modified multiple organ dysfunction score. Organ system failure was defined as a component score of >/=3 on any day during the patient's intensive care unit stay. One hundred and eighty-five patients (89%) developed dysfunction of at least one non neurologic organ system. Ninety-six organ system failures were identified in 74 patients (35%). Respiratory failure was the most common non-neurologic organ system failure, occurring in 23% of patients, whereas cardiovascular failure occurred in 18%. Eight patients (4%) had failure of the coagulation system. One patient had renal failure, whereas no patient developed hepatic failure. In a multivariate model, non-neurologic organ dysfunction was independently associated with hospital mortality (odds ratio for hospital mortality, 1.63; 95% confidence interval, 1.34, 1.98 for one maximum modified multiple organ dysfunction score point). Non-neurologic organ dysfunction was also independently associated with dichotomized Glasgow Outcome Score, as a measure of neurologic outcome (odds ratio for unfavorable neurologic outcome, 1.53; 95% confidence interval, 1.22, 1.98 for one maximum modified multiple organ dysfunction score point). The timing of the organ dysfunction did not appear to be important in the prediction of outcome. CONCLUSIONS: Non-neurologic organ dysfunction is common in patients with severe traumatic brain injury and is independently associated with worse outcome. PMID- 15753761 TI - Analysis of the brain bioavailability of peripherally administered magnesium sulfate: A study in humans with acute brain injury undergoing prolonged induced hypermagnesemia. AB - OBJECTIVE: Based on preclinical investigations, magnesium sulfate (MgSO4) has gained interest as a neuroprotective agent. However, the ability of peripherally administered MgSO4 to penetrate the blood-brain barrier is limited in normal brain. The current study measured the passage of intravenously administered Mg into cerebrospinal fluid in patients with brain injury requiring ventricular drainage. DESIGN: A prospective evaluation of the cerebrospinal fluid total and ionized magnesium concentration, [Mg], during sustained hypermagnesemia was performed. SETTING: Neurosciences intensive care unit at a major teaching institution. PATIENTS: Thirty patients with acute brain injury secondary to subarachnoid hemorrhage, traumatic brain injury, primary intracerebral hemorrhage, subdural hematoma, brain tumor, central nervous system infection, or ischemic stroke were studied. INTERVENTIONS: Patients underwent 24 hrs of induced hypermagnesemia during which total and ionized cerebrospinal fluid [Mg] was measured. Serum [Mg] was adjusted to 2.1-2.5 mmol/L. Cerebrospinal fluid [Mg] was measured at baseline, at 12 and 24 hrs after onset of infusion, and at 12 hrs following infusion termination. MEASUREMENTS AND MAIN RESULTS: At baseline, total (1.25 +/- 0.14 mmol/L) and ionized (0.80 +/- 0.10 mmol/L) cerebrospinal fluid [Mg] was greater than serum total (0.92 +/- 0.18 mmol/L) and ionized (0.63 +/- 0.07 mmol/L) [Mg] (p < .05). Total (1.43 +/- 0.13 mmol/L) and ionized (0.89 +/- 0.12 mmol/L) cerebrospinal fluid [Mg] was maximally increased by 15% and 11% relative to baseline, respectively, during induced hypermagnesemia (p < .05). CONCLUSIONS: Hypermagnesemia produced only marginal increases in total and ionized cerebrospinal fluid [Mg]. Regulation of cerebrospinal fluid [Mg] is largely maintained following acute brain injury and limits the brain bioavailability of MgSO4. PMID- 15753762 TI - TOOLS (treatment with oscillation and open lung strategy) are welcome: timely intervention, combining therapies, strict algorithms. PMID- 15753763 TI - Inotropic options for postresuscitation myocardial dysfunction. PMID- 15753764 TI - Redefining the roles of apoptosis pathways during sepsis. PMID- 15753766 TI - Hyperglycemia, asymmetric dimethylarginine, and patient survival: Dysregulation of complex networks and the metabolic basis of disease. PMID- 15753765 TI - Avoiding a blood transfusion: How much is it worth? PMID- 15753767 TI - The meaning and measurement of physiologic variability? PMID- 15753768 TI - Intensive care or end-of-life care for critically ill cancer patients? PMID- 15753769 TI - "Smart" pumps: A cautionary tale of human factors engineering. PMID- 15753770 TI - It's all in the charge ... PMID- 15753771 TI - Should high mobility group box-1 protein (HMGB1) be measured in patients with severe sepsis and septic shock? If so, when, where, and how? PMID- 15753772 TI - A looming crisis in demand for intensive care unit resources? PMID- 15753773 TI - The Bispectral Index: expanded performance for everyday use in the intensive care unit? PMID- 15753774 TI - General anesthesia in the intensive care unit? Is it ready for "prime time"? PMID- 15753775 TI - Sepsis and intracellular calcium homeostasis, a sparkling story. PMID- 15753776 TI - One more piece in the septic myocardial depression puzzle? PMID- 15753777 TI - Nebulized nitric oxide/nucleophile adduct: Donating nitric oxide to the lung. PMID- 15753778 TI - Calpain: A new target in pulmonary hypertension? PMID- 15753779 TI - The prospect of hemoglobin-based blood substitutes: Still a long stony road to go. PMID- 15753780 TI - Can single nucleotide polymorphisms in innate immune receptors predict development of septic complications in intensive care unit patients? PMID- 15753781 TI - When the brain is injured, the body suffers. PMID- 15753782 TI - Magnesium sulfate therapy in patients with acute neuronal damage: The problem of intravenous administration. PMID- 15753784 TI - The complexity of understanding the immunology of sepsis. PMID- 15753785 TI - Mixing up all the results. PMID- 15753786 TI - Critical care physicians are proponents of early, aggressive, multifaceted therapy. PMID- 15753788 TI - The role of SPECT imaging in the diagnosis of vertebrobasilar insufficiency. PMID- 15753789 TI - Initial application of respiratory-gated 201Tl SPECT in pulmonary malignant tumours. AB - AIM: Respiratory-gated thallium-201 chloride (201Tl) single photon emission computed tomography (SPECT) was used in preliminary investigations to reduce the adverse respiratory motion effects observed on standard ungated SPECT images and to obtain reliable fusion images with computed tomography (CT) in patients with malignant lung tumours. METHODS: Fifteen patients with primary lung cancer (n=10) or metastatic lung tumours (n=5) underwent gated SPECT 20 min after intravenous injection of 148 MBq 201Tl, using triple-headed SPECT and laser light respiratory tracking units. Projection data were acquired by a step and shoot mode, with 20 stops over 120 degrees for each detector and a preset time of 30 s for each 6 degrees stop. Gated end-inspiratory and ungated images were obtained from 1/8 data centred at peak inspiration for each regular respiratory cycle and for the full respiratory cycle data, respectively. The degree and size of tumour 201Tl uptake were compared between these images by regions of interest (ROI) analysis. Gated SPECT images were registered with rest inspiratory CT images using an automated three-dimensional (3D) image registration tool. Registration mismatch was assessed by measuring the 3D distance of the centroid of 14 201Tl-avid peripheral tumours. Attenuation correction of gated SPECT images was performed using CT attenuation values of these fusion images. RESULTS: Gated SPECT images improved image clarity and contrast of tumour 201Tl uptakes compared with ungated images, regardless of the decreased count density due to the use of gated images. The lesion-to-normal (L/N) lung count ratios and ROI size in 18 well circumscribed 201Tl-avid tumours were significantly higher and smaller on gated images (both P<0.0001). Gated images showed positive 201Tl uptakes in two small peripheral tumours, although negative on ungated images, and demarcated 201Tl avid tumours from adjacent 201Tl-avid lymph node or surrounding focal 201Tl uptakes caused by other pathology, although these were not clearly demarcated on ungated images. On fusion images, gated images yielded a significantly better SPECT-CT matching compared with ungated images (P<0.0001). Fusion images accurately localized 201Tl uptakes of tumour/lymph node and other focal pathological/physiological conditions. Attenuation-corrected gated SPECT images further facilitated the detection of 201Tl uptake in small or deeply located lesions, with significantly increased L/N ratios. CONCLUSION: Gated SPECT images facilitate the detection of tumour 201Tl uptake and provide reliable SPECT-CT fusion images, which contribute to accurate interpretation and attenuation correction of Tl SPECT images. PMID- 15753790 TI - Value of delayed 18F-FDG-PET imaging in the detection of hepatocellular carcinoma. AB - BACKGROUND: 18F-Fluorodeoxyglucose-positron emission tomography (18F-FDG-PET) is a very useful imaging technique and is the best modality for the evaluation of many kinds of tumour. However, in the evaluation of hepatocellular carcinoma (HCC), the diagnostic accuracy of routine 60 min static imaging is not satisfactory. Some authors have suggested that delayed 2 h imaging is a better 18F-FDG-PET protocol for tumour detection. However, the value of delayed 3 h imaging has not been clarified. In this study, we performed delayed 2 h and 3 h imaging on patients with HCC and compared their diagnostic accuracy with standard 60 min imaging. METHODS: Twelve patients with HCC were enrolled in this study. Of these 12 patients, four had not been treated and eight had received transcatheter arterial embolization (TAE) therapy for more than 4 months before the PET study. One hour after injection of 18F-FDG, a whole-body scan was performed. In addition, delayed imaging focusing on the liver was also performed 2 h and 3 h after the injection. The standard uptake value (SUV) was calculated for the tumours in each image. RESULTS: The twelve patients had 16 HCCs. Of the 16 HCCs, nine were detected by 18F-FDG-PET scans based on the 1 h images, whereas 10 HCCs were detected based on the 2 or 3 h images. The diagnostic sensitivity increased from 56.3% on the 1 h image to 62.5% on the 2 and 3 h images. In addition, the mean SUV increased from 3.63 at 1 h to 3.86 at 2 h and 3.99 at 3 h after the injection of 18F-FDG. On the other hand, the mean SUV in the normal liver tissue decreased slightly from 2.38 at 1 h to 2.33 at 2 h and 2.31 at 3 h. The tumour to normal liver tissue (T/N) ratio increased from 1.56 at 1 h to 1.68 at 2 h and 1.75 at 3 h. CONCLUSION: In the evaluation of HCC, delayed 2 and 3 h imaging can detect more lesions than standard 1 h imaging. Imaging at 3 h has a better T/N ratio than imaging at 2 h, but does not increase the diagnostic sensitivity. PMID- 15753791 TI - A positron emission tomography/computed tomography (PET/CT) acquisition protocol for CT radiation dose optimization. AB - BACKGROUND: In current combined positron emission tomography/computed tomography (PET/CT) systems, high-quality CT images not only increase diagnostic value by providing anatomic delineation of hyper- and hypometabolic tissues, but also shorten the acquisition time for attenuation correction compared with standard PET imaging. However, this technique potentially introduces more radiation burden to patients as a result of the higher radiation exposure from CT. METHODS: In this study, the radiation doses delivered from typical germanium-based and CT based transmission scans were measured and compared using an anthropomorphic Rando Alderson phantom with insertions of thermoluminescent dosimeters. Image geometric distortion and quantified uptake values in PET images with different manipulating CT acquisition protocols for attenuation correction were also evaluated. RESULTS: It was found that radiation doses during germanium-based transmission scans were almost negligible, while doses from CT-based transmission scans were significantly higher. Using a lower radiation dose, the CT acquisition protocol did not significantly affect attenuation correction and anatomic delineation in PET. CONCLUSIONS: This study revealed the relation between image information and dose. The current PET/CT imaging acquisition protocol was improved by decreasing the radiation risks without sacrificing the diagnostic values. PMID- 15753792 TI - Successful thyroid tissue ablation as defined by a negative whole-body scan or an undetectable thyroglobulin: a comparative study. AB - BACKGROUND: Successful thyroid tissue ablation of patients with well differentiated thyroid cancer can be defined by a negative whole-body scan (WBS) and/or an undetectable thyroglobulin (Tg). Variables associated with success are poorly understood. Tg measurement, although more sensitive than WBS, has not been firmly established as the sole monitoring method. In a previous study, we retrospectively evaluated the variables associated with scintigraphic success. Ablation dose (AD) was the only variable associated with success (odds ratio (OR): 1.96 per 1.85 GBq increment; 95% confidence interval (CI)=1.11-3.46). OBJECTIVES: (1) To determine if the variables associated with success are the same using Tg. (2) To determine whether Tg measurement can become the sole method for assessing ablation success. METHODS: We performed the analysis using a Tg level <2 ng.ml-1 as a criterion for completed ablation. Data were available from 109 patients. RESULTS: Univariate analysis showed an effect of stage (OR=0.05; 95% CI=0.01-0.23) and female sex (OR=2.8; 95% CI=1.14-6.89). Multivariate analysis demonstrated only stage to be a significant predictor of success. Ablation was successful by both methods in 62/109 patients and it failed by both in 10/109. There were 21 WBS- Tg+ and 16 WBS+ Tg- patients. CONCLUSIONS: Investigation of the variables associated with successful ablation yields different results depending on the definition of success. There was a significant incidence of WBS+ Tg- cases after initial ablation. Until it is firmly established that such patients have a benign course both monitoring methods should be used. PMID- 15753793 TI - The identification of reversible dysfunctional myocardium is influenced by the severity of contractile dysfunction and by the length of follow-up. AB - OBJECTIVES: To evaluate the influence of the severity of regional myocardial dysfunction and of the length of follow-up on the identification of myocardial viability with rest-redistribution Tl single photon emission computed tomography (SPECT) and low-dose dobutamine echocardiography (LDDE). METHODS: Twenty-six patients with chronic coronary artery disease and wall motion abnormalities, candidates for revascularization, were included in this study. All patients underwent, in the same week, Tl SPECT and LDDE for pre-revascularization evaluation of myocardial viability. Reversibility of regional dysfunction was assessed by two-dimensional echocardiography, 40+/-20 days (early follow-up) and 12+/-5 months (late follow-up) after revascularization. RESULTS: In a/dyskinetic segments, Tl SPECT showed similar values of sensitivity (78% vs. 71%, P=NS) and slightly higher values of specificity (43% vs. 18%, P<0.01) compared to hypokinetic segments, in predicting functional recovery at early follow-up. No significant changes were observed in the diagnostic accuracy of Tl SPECT at late follow-up. On the contrary, LDDE provided significantly lower values of sensitivity (56% vs. 94%, P<0.05) and higher values of specificity (73% vs. 9%, P<0.01) in a/dyskinetic compared to hypokinetic segments. Specificity of LDDE in a/dyskinetic segments significantly increased from early (73%) to late follow-up (95%; P<0.05). Similarly, positive predictive value in a/dyskinetic segments significantly increased from early (69%) to late follow-up (96%; P<0.05). CONCLUSIONS: The severity of regional dysfunction and the length of follow-up significantly influence the diagnostic accuracy of LDDE but not of rest redistribution Tl SPECT in the identification of myocardial viability. PMID- 15753794 TI - Repeatability of left ventricular ejection fraction and volume measurement for 99mTc-tetrofosmin gated single photon emission computed tomography (SPECT). AB - OBJECTIVES: This study was carried out to assess the repeatability of left ventricular ejection fraction (EF) and volume values obtained using Cedars-Sinai quantitative gated single photon emission computed tomography (SPECT) (QGS) software and relatively low doses of 400-600 MBq of 99mTc-tetrofosmin. METHODS: Repeatability was assessed in a group of 75 patients, with both normal and reduced EF, who underwent repeat 99mTc-tetrofosmin gated SPECT studies and showed no clinical change in cardiac status. Gated SPECT data were acquired 1 h after injection at rest of 400-600 MBq of 99mTc-tetrofosmin. The standard patient dose was 400 MBq; however, some patients with a weight of >90 kg were given increased doses up to a maximum of 600 MBq. RESULTS: There was good correlation of EF and volumes between the first and repeat measurements, and no significant difference between the mean EF and volumes for both the initial and repeat measurements. Background-corrected counts in the left ventricle were calculated and patients were divided into two groups: one with low counts and one with high counts. The mean difference in EF between the first and repeat measurements was significantly higher for patients in the low count group compared with those in the high count group, but there was no significant change in volume. Similarly, the mean sequential difference in EF was significantly higher for patients with normal EF, but there was no significant difference in volume. CONCLUSIONS: We have demonstrated that EF measured using 99mTc-tetrofosmin gated SPECT is repeatable, particularly for patients with low EF, provided that adequate left ventricular counts are obtained. This will require doses greater than 400 MBq in larger patients. Ventricular volumes calculated using QGS may not be sufficiently repeatable for clinical use. PMID- 15753795 TI - Agreement between four available algorithms to evaluate global systolic left and right ventricular function from tomographic radionuclide ventriculography and comparison with planar imaging. AB - BACKGROUND AND AIM: Left and right ventricular ejection fractions (LVEF and RVEF) and end-diastolic and end-systolic volumes (LVEDV, RVEDV, LVESV and RVESV) can be calculated from tomographic radionuclide ventriculography (TRV). The aim of this study was to validate and compare these parameters obtained using four different TRV software programs (QBS, QUBE, 4D-MSPECT and BP-SPECT). METHODS: LVEF obtained from planar radionuclide ventriculography (PRV) was compared with LVEF obtained from TRV using the four different software programs in 166 patients. Furthermore, ventricular volumes obtained using TRV (QBS, QUBE and 4D-MSPECT) were compared with those obtained using BP-SPECT, the latter being the only method with the validation of ventricular volumes in the literature. RESULTS: The correlation of LVEF between PRV and TRV was good for all methods: 0.81 for QBS, 0.79 for QUBE, 0.71 for 4D-MSPECT and 0.79 for BP-SPECT. The mean differences+/-standard deviation (SD) were 3.16+/-9.88, 10.72+/-10.92, 3.43+/-11.79 and 2.91+/-10.39, respectively. The correlation of RVEF between BP-SPECT and QUBE and QBS was poor: 0.33 and 0.38, respectively. LV volumes calculated using QBS, QUBE and 4D-MSPECT correlated well with those obtained using BP-SPECT (0.98, 0.90 and 0.98, respectively), with mean differences+/-SD of 7.31+/-42.94, -22.09+/-36.07 and 40.55+/-39.36, respectively. RV volumes showed poorer correlation between QBS and BP-SPECT and between QUBE and BP-SPECT (0.82 and 0.57, respectively). CONCLUSION: LVEF calculated using TRV correlates well with that calculated using PRV, but is not interchangeable with the value obtained using PRV. Volume calculations (for left and right ventricle) and RVEF require further validation before they can be used in clinical practice. PMID- 15753796 TI - Mapping of the cardiac sympathetic nervous system by single photon emission tomography with technetium-99m-labelled fluorobenzylpiperidine derivative (99mTc FBPBAT): result of a feasibility study in a porcine model and an initial dosimetric estimation in humans. AB - BACKGROUND: Positron emission tomography (PET) and single photon emission tomography (SPET) offer the most promising tools for the in-vivo assessment of the cardiac autonomic nervous system in humans. However, the clinical application of PET and SPET on a routine basis is severely limited by the lack of widely available selective radiotracers. Technetium-99m-labelled 4-fluorobenzyl-4-(2 mercapto-2-methyl-4-aza-pentyl)-4-(2-mercapto-2-methyl-propylamino)-piperidine (99mTc-FBPBAT) is a recently developed radiotracer which exhibited marked adrenergic affinity in previous investigations in vascular smooth muscle cells and cardiac myocytes, and in rats. In this study, we have verified these findings in a porcine model, and evaluated the potential of SPET with 99mTc-FBPBAT to assess the adrenergic nervous system of the heart. METHODS: Using a SPET camera, scintigraphic evaluations were carried out in pigs following intravenous injection of 99mTc-FBPBAT. The specificity of the cardiac uptake was determined by pharmacological intervention, using alpha- and beta-adrenoceptor antagonists and adrenergic re-uptake blocker. Whole-body kinetic and radiation absorbed doses were estimated from whole-body scintigraphies in two healthy volunteers. RESULTS: 99mTc-FBPBAT-SPET demonstrated a homogeneous distribution of radioactivity in myocardium of pigs and in humans. The cardiac uptake was specifically suppressed by previous treatment of the animals with metoprolol and prazosin, and was displaceable by norepinephrine. In contrast, the inhibition of radioactivity uptake into the heart was less pronounced after pretreatment with desipramine. The peak radioactivity in blood was determined after 1.5-2 min, followed by a plateau of nearly constant radioactivity from 25-30 min onwards. Within 6 h, more than 35% of the injected activity was excreted in the urine. The effective dose according to International Commission on Radiological Protection Publication 60 (ICRP 60) was 0.0064 mSv.MBq-1 for adults. CONCLUSION: In view of these findings, we conclude that the myocardial uptake of 99mTc-FBPBAT reflects the sympathetic adrenergic nervous system of the heart. The effective dose estimated indicates that the clinical use of 99mTc-FBPBAT results in an acceptable radiation dose in humans. Despite the relatively high radioactivity uptake into the lung and liver, 99mTc-FBPBAT appears to be the first promising Tc-based radiotracer for scintigraphic assessment of the cardiac adrenergic system. This result encourages further development of Tc-based agents for routine SPET studies in humans. PMID- 15753798 TI - Molecular-size fractionation of pentastarch, radiolabelling with 99mTc, and evaluation of biological behaviour in mice. AB - BACKGROUND: Pentastarch is used clinically as a plasma volume expander for the management of substantial blood loss. 99mTc labelled pentastarch may be useful as a diagnostic agent in place of 99mTc labelled red blood cells. METHODS: Commercial pentastarch (PS; molecular weight (MW) 240 kDa) was separated according to molecular size by using chromatography, and the fractions were pooled as small (MW 128 kDa), medium (MW 277 kDa) and large (MW 510 kDa) pentastarch. We studied the effect of various physicochemical parameters on the efficiency of radiolabelling with 99mTc and on the stability of the products, and evaluated the biological properties of the 99mTc labelled preparations. RESULTS: We developed an optimised kit formulation containing 3.25 mg pentastarch and 0.13 mg gentisic acid that can be reliably labelled with 99mTc at pH 6.6-8.2 with good stability. In mice, the 99mTc labelled medium pentastarch showed the more favourable blood retention properties (56% of initial blood activity is retained after 3 h) with lower liver levels. PMID- 15753797 TI - Radiolabelling of egg meals for gastric emptying studies: a comparison of 99mTc sulfur and 99mTc stannous colloids. AB - OBJECTIVES: To establish whether 99mTc stannous (Tc-Sn) colloid is a suitable alternative to 99mTc sulfur (Tc-S) colloid for gastrointestinal studies, we compared the per cent binding to egg solids (%BS) and radiochemical purity (RCP) of both colloids in digesting media. METHODS: Egg white and yolk containing colloids were cooked separately and mashed. Samples of 4-5 g were digested over 2 3 h (37 degrees C) in excess simulated gastric juice (SGJ: 15 ml of 0.1 M HCl and 0.5 g.l-1 pepsin) or water, centrifuged, imaged with a gamma camera and the %BS computed. RCP was determined in aspirates taken from these preparations and from solutions of colloid added directly to SGJ. RESULTS: The %BS in egg white after 3 h in SGJ for both colloids were similar: Tc-Sn, 62+/-8 (n=12); Tc-S, 61+/-6 (n=8), but markedly lower than 95% (the often quoted literature value). Egg yolk was digested more rapidly than egg white: %BS after 2 h in SGJ for Tc-Sn colloid was 55+/-10 (n=5) in the yolk, compared to 77+/-4 (n=5) in the white (P<0.01). The RCP for Tc-S colloid alone in SGJ was >94% over 3 h but for Tc-Sn colloid was as low as 14%. For egg white, the RCP in SGJ was 91-96% for Tc-S and 80-91% for Tc-Sn. For egg yolk the RCPs in SGJ were similar for both colloids (>90%). The RCP in water digesting egg white or yolk containing either colloid was always lower than in the corresponding SGJ aspirates, indicating a leakage of small amounts of non-colloidal 99mTc. CONCLUSIONS: Although 99mTc-Sn almost completely dissociates in SGJ, once cooked in egg it is digested similarly to 99mTc-S. Variations in the size of digesting egg fragments and in SGJ composition can reconcile the lower %BS values obtained with previously reported higher results. 99mTc-Sn colloid cooked in egg appears suitable for gastrointestinal studies. PMID- 15753799 TI - A cost-effective and versatile xenon gas dispenser. AB - OBJECTIVE: To modify a commercial xenon gas dispenser so that two xenon unit-dose vials could be combined with a modified dispenser to deliver a recommended dose. METHOD: To maintain the same operating mechanism, changes were made only to the vial shield and the needle port of the original gas dispenser. The modified gas dispenser consisted of two puncture needles and two vial holders shielded with the same thickness of lead as the commercial dispenser. RESULTS: Our evaluation showed that the modified gas dispenser operated the same way as the commercial unit, and the average 133Xe residual activity in either one or two xenon unit dose vials of the modified gas dispenser was not significantly different from that in one vial of the commercial xenon gas dispenser. CONCLUSION: The modified xenon gas dispenser allows the stock of xenon gas vials to be managed cost effectively. The modified unit can be used to dispense two low-activity xenon gas vials to deliver a standard dose to a patient. Also, the modified gas dispenser can be used to combine different amounts of xenon activity in two unit-dose vials in order to customize the dose delivered to patients with special needs (e.g., obese patients). Our modified device can also function as a single-dose dispenser by placing an empty vial alongside the unit-dose vial of radioactive xenon gas. PMID- 15753809 TI - Osteoporosis. PMID- 15753808 TI - Duloxetine fights major depressive disorder. PMID- 15753810 TI - Recognizing multiple myeloma. PMID- 15753812 TI - Otitis media: new information on an old disease. PMID- 15753815 TI - Health literacy a barrier to patient education. PMID- 15753814 TI - Influence positive change with motivational interviewing. PMID- 15753816 TI - An update on cholesterol guidelines. PMID- 15753817 TI - NPs vs. other healthcare providers in end-of-life care. PMID- 15753828 TI - Revised information for lovenox. PMID- 15753818 TI - Making headlines. PMID- 15753829 TI - Drug treats two sources of cholesterol. PMID- 15753831 TI - FDA: antidepressants a risk for kids. PMID- 15753830 TI - Finasteride with doxazosin to reduce BPH progression. PMID- 15753833 TI - The volume-outcome relationship in pediatric intensive care units: a conceptual advance or a statistical novelty? PMID- 15753834 TI - Evaluating hyperosmolar therapy for hyperosmolar disease. PMID- 15753835 TI - Treatment of pleural empyemas. PMID- 15753836 TI - "Extrasystoles" during kangaroo care. PMID- 15753838 TI - The role of postischemic reperfusion injury and other nonantigen-dependent inflammatory pathways in transplantation. AB - The Injury Hypothesis, first published in 1994 and modified several times between 1996 and 2002, holds that the reactive oxygen species-mediated reperfusion injury to allografts initiates and induces the alloimmune response and contributes to alloatherogenesis. Recent experimental and clinical evidence in support of the concept is presented suggesting that (1) reactive oxygen species-mediated allograft injury activates the innate immune system of the donor and recipient; (2) injury-induced putative endogenous ligands of Toll-like receptors (TLRs) of host origin such as heat shock proteins interact with and activate TLR4-bearing dendritic cells that mature and induce the adaptive alloimmune response (acute rejection), and interact with and activate TLR4-bearing vascular cells contributing to the development of alloatherosclerosis (chronic rejection); and (3) TLR4-triggered signaling, involved in the establishment of a reperfusion injury, seems to use myeloid differentiation marker 88-independent, Toll/interleukin-1 receptor domain containing adaptor inducing interferon-beta dependent pathways that are associated with the maturation of dendritic cells and induction of interferon-inducible genes. PMID- 15753839 TI - Nondirected living donation: a survey of current trends and practices. AB - The use of living donors in general is increasing, and there is evidence of increasing willingness among transplant centers to consider nondirected living donation (NDLD) as well. Yet few systematic data are available on the number of centers performing NDLD and the degree to which current policies and practices reflect existing guideline recommendations. We conducted a telephone survey of the 25 highest volume transplant centers and 25 highest volume organ procurement organizations (OPOs) to describe their NDLD-related policies and experiences. Survey responses revealed that inquiries about NDLD from interested potential donors are common, likely to increase, and generally not formally monitored. More than half of transplant centers surveyed currently perform or are willing to perform NDLD (14/25), and although only three OPOs currently facilitate NDLD, the majority of the remaining organizations reported that an NDLD policy was in development. Among those organizations that did facilitate NDLD, the majority applied current living-donor guidelines and practices to NDLDs. Most transplant centers performing NDLD distributed these donations to patients from their own waiting list by using United Network for Organ Sharing criteria. This preliminary description of current NDLD experiences suggests that although NDLD is growing, the policies and practices governing it are being developed on an ad hoc basis at the level of individual organizations. Additional research is needed to characterize the full range of NDLD activity at both higher and lower volume organizations, and to explore a broader range of questions about current NDLD policy and practice in the United States. PMID- 15753840 TI - Induction and maintenance of T-dependent or T-independent xenotolerance by nonprimarily-vascularized skin or thymus grafts. AB - BACKGROUND: The success of clinical xenotransplantation will depend on induction of xenotolerance. We have previously shown that combined xenothymus and vascularized xenoheart transplantation under the coverage of a tolerizing regimen (TR) can induce and maintain full xenotolerance. Here, induction/maintenance of xenotolerance using nonprimarily-vascularized thymus and/or skin grafts was investigated. MATERIALS AND METHODS: Hamster skin or thymus or combined skin and thymus transplantation was performed in nude rat recipients with or without administering a TR (NK cell depletion, day -14; xenoantigen infusion, day -14; Leflunomide, day -14 through +14). Xenotolerance was confirmed by subsequent transplantation of a vascularized hamster heart, measurement of xenoantibody formation, or mixed lymphocyte reaction (MLR). RESULTS: Skin grafts were as effective as vascularized heart grafts to induce/maintain T-independent xenotolerance. Even without TR and despite being rejected themselves, xenoskin grafts lead to progressively developing xenononreactivity. Xenothymus transplantation induced xenotolerance in the T-dependent but not in the T independent immune compartment, leading to rejection of subsequently transplanted hamster hearts by T-independent mechanisms (production of IgM but not IgG xenoantibodies (Xabs), presence of antihamster MLR nonresponsiveness). Combined skin and thymus xenotransplantation sensitized the T-cell compartment, leading to hyperacute rejection of subsequently transplanted hamster hearts. This was not the case when the skin grafts were transplanted late (2 months) after the thymus grafts. CONCLUSIONS: Xenogeneic skin and xenogeneic thymus grafts have opposite xenotolerance inducing capacities in the T-independent as compared to the T dependent immune compartment. Thymus grafts induce and maintain T-dependent but not T-independent xenotolerance. Skin grafts alone induce T-independent xenotolerance but sensitize the T-cell compartment when transplanted concomitantly with thymus grafts. PMID- 15753841 TI - Human amniotic mesenchymal cells have some characteristics of cardiomyocytes. AB - BACKGROUND: Cellular cardiomyoplasty (CCM) is a major method for the treatment of heart failure because adult cardiomyocytes do not regenerate after ischemic injury, which results in heart failure. There is a great deal of interest in finding suitable new cell sources for use in CCM. Here, we report that human amniotic mesenchymal cells (hAMC), which are multipotent cells derived from fetal mesoderm, may be a suitable cell source for CCM. METHODS: Freshly isolated hAMC were examined to detect the expression of cardiac-specific genes by reverse transcription polymerase chain reaction and immunocytochemistry. hAMC were cocultivated with neonatal rat heart explants and transplanted into myocardial infarcts in the rat heart. RESULTS: hAMC expressed cardiac-specific transcription factor GATA4, cardiac-specific genes, such as myosin light chain (MLC)-2a, MLC 2v, cTnI, and cTnT, and the alpha-subunits of the cardiac-specific L-type calcium channel (alpha1c) and the transient outward potassium channel (Kv4.3). After stimulation with basic fibroblast growth factor (bFGF) or activin A, hAMC expressed Nkx2.5, a specific transcription factor for the cardiomyocyte and cardiac-specific marker atrial natriuretic peptide. In addition, the cardiac specific gene alpha-myosin heavy chain was detected after treatment with activin A. Coculture experiments confirmed that hAMC were able to both integrate into cardiac tissues and differentiate into cardiomyocyte-like cells. After transplantation into the myocardial infarcts in rat hearts, hAMC survived in the scar tissue for at least 2 months and differentiated into cardiomyocyte-like cells. CONCLUSION: The results of the present study suggest that hAMC possess some characteristics of cardiomyocytes. PMID- 15753842 TI - A role for CD54 (intercellular adhesion molecule-1) in leukocyte recruitment to the lung during the development of experimental idiopathic pneumonia syndrome. AB - BACKGROUND: Idiopathic pneumonia syndrome (IPS) is a frequently fatal complication of allogeneic bone marrow transplantation (BMT). IPS is associated with elevated bronchoalveolar lavage (BAL) fluid levels of tumor necrosis factor alpha and lipopolysaccharide, both of which are potent activators of endothelial cells (ECs). EC expression of the adhesion molecule CD54 (intercellular adhesion molecule [ICAM]-1) has been shown to be a major regulator of pulmonary inflammation in various experimental models. METHODS: Using a well-established murine BMT system in which lung injury and graft-versus-host disease (GvHD) are induced by minor histocompatibility antigenic differences between donor and host, the RNase Protection Assay, mice deficient in ICAM-1 expression, and a monoclonal blocking antibody to ICAM, we evaluated the role of the pulmonary vascular expression of CD54 in the development of IPS. RESULTS: Enhanced pulmonary vascular expression of ICAM-1 coincided with the development of IPS. When ICAM-1 /- mice were used as allogeneic BMT recipients, IPS severity (measured by lung histopathology, BAL cellularity, and cytokine expression) was significantly reduced compared with wild-type controls. Similar results were also observed when wild-type recipients were treated with a monoclonal blocking antibody to ICAM-1. Surprisingly, ICAM-1 had differential effects on leukocyte infiltration into GvHD target organs; ICAM-1 deficiency had no impact on intestinal histopathology, whereas ICAM-1-/- BMT recipients had significantly enhanced hepatic injury. CONCLUSIONS: These data demonstrate that although the expression of ICAM-1 is critical for the development of IPS, different mechanisms of leukocyte recruitment are operative in other GvHD target organs. PMID- 15753843 TI - Macrophage depletion prolongs discordant but not concordant islet xenograft survival. AB - BACKGROUND: T cells and macrophages play a major role in the rejection of xenografted islets. Depending on the phylogenetic disparity, direct or indirect antigen presentation is predominant. The aim of this study was to analyze in vitro the predominance of direct or indirect presentation by depleting antigen presenting cells in concordant and discordant xenogeneic combinations. In vivo, we analyzed the effect of macrophage depletion on concordant and discordant islet xenograft survival to assess in which combination this strategy can be used as therapeutic tool. MATERIALS AND METHODS: In vitro, we performed mouse anti-rat and anti-human mixed lymphocyte reactions (MLR) after depletion of responder or stimulator antigen-presenting cells by magnetic sorting. In vivo, streptozotocin induced diabetic C57BL/6 mice were treated by gadolinium chloride (GdCl) to deplete macrophages, and rat or human islets were transplanted under the kidney capsule. Islet function was followed by glycemia, and xenografts were analyzed at regular intervals for histology and immunohistochemistry. RESULTS: Mouse anti-rat MLR showed a predominant direct antigen presentation pathway, whereas in mouse anti-human MLR, direct and indirect pathways were similarly involved. Survival of rat islets was not modified by GdCl therapy. In contrast, survival of human islets was significantly prolonged in GdCl-treated mice. Macrophage infiltration was decreased in concordant and discordant GdCl-treated xenografts at day 4 compared with controls. At day 15, macrophage, CD4+, and CD8+ cell infiltration was similar in all groups. CONCLUSIONS: Our results indicate that direct antigen presentation is dominant in the rejection mechanism of concordant islet xenografts and cannot be influenced by host macrophage depletion. Both direct and indirect antigen presentation are involved in rejection of discordant xenogeneic islets. Macrophage depletion should only be considered as therapeutic tool for discordant islet xenotransplantation. PMID- 15753844 TI - Enrichment of hepatocytes differentiated from mouse embryonic stem cells as a transplantable source. AB - BACKGROUND: We previously reported that hepatocytes can be differentiated from embryonic stem (ES) cells by way of embryoid body (EB) formation and are transplantable into the mouse liver. However, the transplantation of EB-derived cells frequently resulted in teratoma formation in the recipient liver. In the present study, we eliminated the tumorigenic cells from EB outgrowths and examined the effects of enriched ES-cell-derived hepatocyte transplantation into an injured liver. METHODS: On day 15 in culture, the EBs were partially disaggregated and subcultured. Hepatocytes in the subcultured cells were examined by the expression of hepatocyte markers. Undifferentiated cells contaminating in the EB-derived cells were eliminated by Percoll discontinuous gradient centrifugation. Furthermore, undifferentiated cells, endothelial cells, and macrophages were eliminated by magnetic cell sorting using platelet/endothelial cell adhesion molecule (PECAM)-1 and Mac-1 antibodies. These enriched ES-cell derived hepatocytes were then transplanted into the injured mouse liver. RESULTS: Percoll centrifugation and PECAM-1 antibodies eliminated the undifferentiated cells expressing Oct-3/4 from the EB-derived cells. ES-cell-derived hepatocytes showed expression of liver-related genes, synthesis of urea and glycogen, and structural characteristics during subculture. A transplantation study showed that the enriched ES-cell-derived hepatocytes integrated into the injured mouse liver and produced no teratomas. When the ES-cell-derived hepatocytes were transplanted into a CCl4-injured liver, the liver function was subsequently improved. CONCLUSIONS: Functional hepatocytes can be differentiated from mouse ES cells by way of EB formation. The elimination of undifferentiated cells from the EBs provides transplantable cells for liver failure without tumorigenicity. PMID- 15753845 TI - Development and maintenance of donor-specific chimerism in semi-allogenic and fully major histocompatibility complex mismatched facial allograft transplants. AB - BACKGROUND: The clinical application of composite tissue allograft transplants opened the discussion on the restoration of facial deformities by allotransplantation. We introduce a hemifacial allograft transplant model to investigate the rationale for the development of operational tolerance across a major histocompatibility complex (MHC) barrier. MATERIAL AND METHODS: Thirty rats were studied in five groups of six animals each. The composite hemiface isograft transplantations were performed in group 1. Allograft rejection controls included semi-allogenic transplantations from LBN (RT1(1+n) donors (group 2) and fully allogenic transplantations from ACI (RT1a) donors (group 3) to LEW (RT1(1)) recipients. In the allograft treatment groups, recipients of LBN (group 4) and ACI donors (group 5) were treated with cyclosporine A monotherapy (16 mg/kg/day, tapered to 2 mg/kg/day). Face allografts were evaluated clinically and histologically. Donor-specific chimerism for MHC class I RT1n and RT1a antigens was assessed by flow cytometry. Mixed lymphocyte reaction for donor-specific tolerance in vitro was tested at day 160 posttransplant. RESULTS: Isograft controls survived indefinitely. All nontreated allografts rejected within 5 to 8 days posttransplant. Long-term survival was achieved in 100% of LBN (up to 400 days) and ACI (up to 330 days) recipients. At day 160, posttransplant donor specific chimerism was present in recipients of LBN (10.14% CD4/RT1n, 6.38% CD8/RT1n, 10.02% CD45RA/RT1n) and ACI (17.54% CD4/RT1a, 9.28% CD8/RT1a) transplants, and mixed lymphocyte reaction confirmed tolerance in recipients of LBN transplants and moderate reactivity in recipients of ACI allografts. CONCLUSION: Operational tolerance was induced in hemiface allograft transplants across an MHC barrier under cyclosporine A monotherapy protocol. It was associated directly with the presence of multilineage donor-specific chimerism. PMID- 15753846 TI - Interleukin-10 but not transforming growth factor-beta is essential for generation and suppressor function of regulatory cells induced by intratracheal delivery of alloantigen. AB - BACKGROUND: We previously reported that intratracheal delivery of alloantigen induced regulatory cells in mouse heart-transplantation model. Here, we investigated roles of interleukin (IL)-10 and transforming growth factor (TGF) beta in induction and effector phases of the regulatory cells. METHODS: CBA mice were pretreated with intratracheal delivery of C57BL/10 splenocytes and administration of neutralizing anti-IL-10 or anti-TGF-beta monoclonal antibody (mAb). Seven days after the pretreatment, naive CBA mice (secondary recipients) were given adoptive transfer of splenocytes from the pretreated mice and underwent heart grafting from C57BL/10 mice. To determine roles of these cytokines in the effector phase of the regulatory cells, anti-IL-10 or anti-TGF beta mAb was administered weekly into the secondary recipients after the adoptive transfer. RESULTS: Adoptive transfer of splenocytes from CBA mice that had been pretreated with intratracheal delivery of C57BL/10 splenocytes significantly prolonged the survival of C57BL/10 allograft (median survival time [MST] 68 days) as compared with adoptive transfer from untreated CBA mice (MST 12 days). In the induction phase, anti-IL-10 mAb abrogated development of the regulatory cells that afforded prolonged allograft survival in the secondary recipients (MST 20 days), whereas anti-TGF-beta mAb did not abrogate it (MST 88 days). In the effector phase, anti-IL-10 mAb abrogated prolonged allograft survival afforded by adoptive transfer of the regulatory cells in the secondary recipients (MST 27 days), whereas anti-TGF-beta mAb did not abrogate suppressor function of the regulatory cells (MST 53 days). CONCLUSION: IL-10 but not TGF-beta was required for generation and suppressor function of the regulatory cells induced by intratracheal delivery of alloantigen. PMID- 15753847 TI - Efficacy and safety outcomes among de novo renal transplant recipients managed by C2 monitoring of cyclosporine a microemulsion: results of a 12-month, randomized, multicenter study. AB - BACKGROUND: The clinical benefits of C2 monitoring of cyclosporine microemulsion have been demonstrated, but C2 targets in renal transplant recipients during the first year require validation. METHODS: MO2ART was a prospective, multicenter study of renal transplant recipients managed by C2 monitoring of cyclosporine microemulsion with steroids and mycophenolate mofetil or azathioprine. Patients were randomized on day 3 to two groups, which were managed from month 3 with higher or lower C2 target ranges (months 4-6, 1,000-1,200 ng/mL vs. 800-1,000 ng/mL; months 7-12, 800-1,000 ng/mL vs. 600-800 ng/mL, respectively). The primary endpoint was the glomerular filtration rate (GFR) at month 12. RESULTS: A total of 296 patients were recruited, of whom 250 remained in the study at 3 months (higher-C2, n=131; lower-C2, n=119). GFR at 12 months did not differ between the higher- and lower-C2 groups (65+/-17 mL/min vs. 66+/-14 mL/min). When patients were regrouped according to C2 achieved by months 8 to 12, those with the lowest C2 (<700 ng/mL) showed the lowest GFR at month 3 and the most pronounced increase in GFR between month 3 and month 12 (P=0.04). Five episodes of biopsy-proven acute rejection occurred after month 3 (higher-C2 group, n=2; lower-C2 group, n=3). The overall 12-month Kaplan-Meier incidence of biopsy-proven acute rejection was 13.7%. Patient and graft survival were 93% and 89%, respectively, at 12 months. CONCLUSION: Both C2 target ranges investigated showed excellent and nearly equivalent outcomes at 12 months. The decision to target the higher or lower end of these C2 ranges should be made on an individual basis, taking into account patient and graft characteristics, and co-medication. PMID- 15753848 TI - Sequential expression of three known protective genes in cardiac biopsies after transplantation. AB - BACKGROUND: The expression of the "protective" genes A20, heme oxygenase (HO)-1, and Bcl-xl in rodent allografts and xenografts correlates with long-term survival of transplanted hearts. We investigated the expression of HO-1, Bcl-2, and A20 in sequential biopsies from nine cardiac transplant recipients by using quantitative real-time reverse-transcriptase polymerase chain reaction and immunohistochemistry. METHODS: Five to 16 endomyocardial biopsies were analyzed from each patient 7 to 365 days after transplantation. Biopsies were classified as acute rejection (AR) by International Society of Heart and Lung Transplantation criteria. mRNA values were normalized against an endogenous control gene (18S), and protein expression was analyzed by immunohistochemistry. RESULTS: All genes were expressed at every time point. HO-1 was significantly higher in the first 2 months (2 months vs. 10+ months, P<0.05) and was associated with AR (0.30+/-0.07) versus nonrejection (0.16+/-0.02, P=0.026). In contrast, expression of Bcl-2 and A20 was low at 2 months, but both increased with time (P<0.05, 2 months vs. 10+ months for Bcl-2 and A20). There was no significant association of Bcl-2 or A20 with AR. Immunocytochemistry revealed that HO-1 localizes to infiltrating cells and not parenchymal cells in cardiac biopsies. In contrast, Bcl-2 and A20 were found to localize to endothelial, smooth muscle, and infiltrating cells. CONCLUSIONS: HO-1 is induced early after transplantation, whereas Bcl-2 and A20 seem to be induced as part of the chronic response. These differences together with different localization sites in vivo suggest they have different roles in protection from injury after cardiac transplantation. PMID- 15753849 TI - Development of posttransplant antidonor HLA antibodies is associated with acute humoral rejection and early graft dysfunction. AB - BACKGROUND: The goal of this study was to determine whether the production of posttransplant antibodies directed against donor HLA mismatches (donor specific antibody; DSA) is associated with renal allograft rejection and early graft dysfunction. METHODS: Forty-nine adult renal allograft recipients with increased risk of rejection were enrolled during the period of October 2001 through May 2003 and were prospectively monitored for the development of anti-HLA antibodies. RESULTS: Of 49 patients, eight (16.3 %) patients were diagnosed with acute humoral rejection (AHR) and 11/49 (22.4%) patients were diagnosed with acute cellular rejection (ACR). A strong association between pretransplant HLA sensitization and AHR was found (P=0.005). Of the eight patients diagnosed with AHR, the majority developed DSA before or concomitant with episodes of rejection (P<0.001). Only 3 of 41 patients (7.3%) without AHR developed DSA. The pathogenic role of alloantibodies was further substantiated by analyzing their association with graft function as measured by serum creatinine levels. The average serum creatinine after the third month posttransplantation in DSA producers was 2.24+/ 1.01 mg/dL, while in non-DSA patients the average serum creatinine was 1.41+/ 0.37 mg/dL (P<0.01). CONCLUSION: This study reveals a strong association between the production of DSA, AHR, and early graft dysfunction. Our findings indicate that prospective monitoring for anti-HLA antibodies following transplantation is a useful test for the diagnosis and classification of AHR for identifying patients at risk of early graft dysfunction. PMID- 15753850 TI - Evaluation of pretransplant immunologic status in kidney-transplant recipients by panel reactive antibody and soluble CD30 determinations. AB - OBJECTIVES: To retrospectively compare the accuracy of pretransplant panel of reactivity antibodies (PRA) and serum level of soluble CD30 (sCD30) in predicting early (< 6 months) acute rejection (AR) in living-donor and deceased-donor kidney transplant (KT) patients. METHODS: Pretransplant sera of 24 KT recipients were retrospectively tested for sCD30 and compared with PRA. Inclusion criteria were de novo graft patients on calcineurin-inhibitor-based immunosuppression, minimum follow-up of 1 year, alive with a functioning graft, and stable renal function over the last 12 months. Objective measures were incidence of biopsy-proven AR (BPAR) within 6 months of KT and sCD30 and PRA diagnostic indexes. The relative risk (RR) of BPAR for each test was also obtained. RESULTS: Fourteen (58.3%) patients presented at least one episode of BPAR within 6 months of KT. All rejection episodes were responsive to steroid treatment. PRA was positive in six (25%) patients, and four (66.7%) of them presented at least one episode of BPAR. sCD30 tested positive in nine (37.5%) patients, and all these later presented at least one episode of BPAR. sCD30 and PRA diagnostic indexes in predicting early (< 6 months) BPAR were sensitivity 64.2% versus 28.5%; specificity 100% versus 80%; accuracy 79.1% versus 50%; positive predictive value 100% versus 66.6%; and negative predictive value 66.6% versus 44.4%. The RR of early AR was 1.4 in PRA positive patients and extremely higher in the sCD30-positive group. CONCLUSIONS: Pretransplant sCD30 is a more accurate predictor of AR when compared with PRA. These results support its use in the pretransplant work-up of kidney-graft recipients. PMID- 15753851 TI - Donor CD31 genotype impacts on transplant complications after human leukocyte antigen-matched sibling allogeneic bone marrow transplantation. AB - Mismatch for the adhesion molecule CD31 (PECAM-1) has been associated in some studies with graft-versus-host disease (GVHD), suggesting a role for CD31 as a minor histocompatibility antigen. We examined polymorphisms of the CD31 (PECAM-1) gene in 74 patients and their human leukocyte antigen-matched sibling donors, comparing CD31 genotype with outcomes of occurrence of GVHD and survival using regression analysis. Polymorphisms in codon 125, 563, and 670 are strongly linked forming conserved haplotypes. Donor CD31 (val/asn/gly) haplotype was associated with acute GVHD (P=0.004, odds ratio 7.5). In addition, donor heterozygosity at codon 563 was significantly associated with worse overall survival after correcting for other known variables by regression modeling. Peptide binding predictions support the hypothesis that CD31 could act as a minor histocompatibility antigen. Assessment for CD31 gene status may be of value in pretransplant assessment of bone marrow transplant recipients and donors for prediction of likely transplant-related complications. PMID- 15753852 TI - Omega-3 fatty acids improve hepatic steatosis in a murine model: potential implications for the marginal steatotic liver donor. AB - The presence of nonalcoholic fatty liver disease often precludes potential organs from being used for transplantation. To date, there is no adequate treatment for hepatic steatosis, and it is expected that, because of increased obesity in Western society, the incidence of this disorder will increase. We investigated the effect of omega-3 polyunsaturated fatty acid supplementation on the treatment of hepatic steatosis in C57/Bl6 mice fed a high-carbohydrate, fat-free diet and in B6.V-Lep(ob) obese mice. Omega-3 fatty acid supplementation reversed hepatic steatosis in C57/Bl6 mice fed a high-carbohydrate, fat-free diet and converted macrovesicular to microvesicular steatosis in B6.V-Lep(ob) obese mice as determined by histology, magnetic resonance spectroscopy, and liver biochemistry We therefore conclude that omega-3 fatty acid supplementation improves hepatic steatosis in mice and may be used to increase the pool of potential live liver donors that are currently excluded because of the presence of macrovesicular steatosis. PMID- 15753853 TI - Early postnephrectomy donor renal function: laparoscopic versus open procedure. AB - Laparoscopic donor nephrectomy (LDN) is becoming the method of choice to procure kidneys from living donors. Despite the benefits to the donor, there have been concerns over the transient deterioration of renal function in the recipient of LDN compared with standard nephrectomy. We carried out a retrospective review of all living donors at our institution between January 2000 and December 2002. On the first postoperative day, the fall in renal function in laparoscopic donors is significantly greater than the fall seen in open donors. This difference could not be explained by relative hypotension, excessive blood loss, or inadequate fluid replacement in the laparoscopic group. Importantly, this difference is no longer evident by the third postoperative day. We speculate that this may be secondary to the pneumoperitoneum or the prolonged anesthesia on glomerular filtration rate. Furthermore, this finding could explain the slower recovery of graft function in recipients of laparoscopically procured kidney transplants. PMID- 15753854 TI - Increased vitamin A and E levels in adult cystic fibrosis patients after lung transplantation. AB - Most patients with cystic fibrosis (CF) and pancreatic insufficiency require supplementation with fat-soluble vitamins to maintain normal serum levels. Even with supplementation, toxicity is rare. We evaluated serum vitamin A and E levels in 23 adult patients with CF who underwent double lung transplantation. Twenty one of the subjects were pancreatic insufficient. Fifteen subjects had serum vitamin levels before and after transplant. The median time posttransplantation for these subjects was 9 months. Mean serum vitamin A and E levels were significantly higher posttransplantation (P<0.0001, P<0.001, respectively). Eight subjects who only had posttransplant vitamin levels also had abnormally high vitamin A levels. Although the etiology of this novel finding is unclear, possibilities include altered absorption, drug interactions, impaired retinol metabolism, or increased hepatic synthesis of retinol binding protein. PMID- 15753855 TI - Progressive disappearance of anti-hepatitis B surface antigen antibody and reverse seroconversion after allogeneic hematopoietic stem cell transplantation in patients with previous hepatitis B virus infection. AB - Reactivation of resolved hepatitis B virus (HBV) infection, which is known as reverse seroconversion (RS), has been reported as a rare complication of allogeneic hematopoietic stem cell transplantation. We retrospectively studied HBV serologic markers in 14 recipients with pretransplant anti-hepatitis B surface antigen antibody (anti-HBs). Progressive decreases in anti-HBs titer were observed in all cases. In 12 cases, anti-HBs titer had decreased to under the protective value. RS occurred in seven cases after disappearance of anti-HBs. Although reseroconversion occurred in five cases, two cases remained in an HBV carrier status after resolution of hepatitis. In the other five cases, RS did not occur even after disappearance of anti-HBs. The actual risks of anti-HBs disappearance and RS were estimated to be 75.0% and 39.8% at 2 years and 100.0% and 70.0% at 5 years, respectively. In conclusion, RS is a late-onset complication with high frequency that can be predicted by careful monitoring of progressive decrease in anti-HBs titer. PMID- 15753856 TI - CD36 (Naka) sensitization with platelet-transfusion refractoriness in a liver transplant recipient. PMID- 15753858 TI - The United Network for Organ Sharing position on using donors with primary central nervous system malignancies. PMID- 15753857 TI - First successful application of side-to-side intrahepatic cholangiojejunostomy to biliary obstruction after living-donor liver transplantation. PMID- 15753859 TI - United network for organ sharing publication on scientific registry of transplant recipients central nervous system donor cancer transmission data. PMID- 15753860 TI - Hemorrhagic cystitis caused by adenovirus type 34 after allogeneic bone marrow transplantation. PMID- 15753861 TI - Small-for-size syndrome after living-donor liver transplantation treated by "portal vein wrapping" and single plasmapheresis. PMID- 15753862 TI - Variations in biliary anatomy associated with trifurcated portal vein in right lobe living-donor liver transplantation. PMID- 15753865 TI - [Application of phase contrast imaging to mammography]. AB - Phase contrast images were obtained experimentally by using a customized mammography unit with a nominal focal spot size of 100 microm and variable source to-image distances of up to 1.5 m. The purpose of this study was to examine the applicability and potential usefulness of phase contrast imaging for mammography. A mammography phantom (ACR156 RMI phantom) was imaged, and its visibility was examined. The optical density of the phantom images was adjusted to approximately 1.3 for both the contact and phase contrast images. Forty-one observers (18 medical doctors and 23 radiological technologists) participated in visual evaluation of the images. Results showed that, in comparison with the images of contact mammography, the phantom images of phase contrast imaging demonstrated statistically significantly superior visibility for fibers, clustered micro calcifications, and masses. Therefore, phase contrast imaging obtained by using the customized mammography unit would be useful for improving diagnostic accuracy in mammography. PMID- 15753866 TI - [Examination of phantom scatter factors in irregular fields with enhanced dynamic wedge]. AB - Use of an enhanced dynamic wedge (EDW) changes the size of the irradiation field in real time because of the shift of the flexible aperture during irradiation. Therefore, when EDW is used, it is speculated that the scatter factors proposed by Khan et al. may differ from that obtained in an open irradiation field. The aim of the present study was to investigate whether the total scatter factors (S(cp)) can also be separated into the collimator scatter factors (S(c)) and phantom scatter factors (S(p)) when using EDW, and whether S(p) in an irregular irradiation field using EDW and one without the use of EDW can likewise be handled. Results indicated that the values of S(cp) and S(c) in a square or rectangular irradiation field using EDW tended to differ from those in an open irradiation field, whereas S(p) was almost the same, with a slight variation of about 0.5%, indicating that S(cp) could be separated into S(c) and S(p). However, for an irregular irradiation field, the variations in S(p) exceeded 1% in some cases irrespective of the use or non-use of EDW. Depending on the size and geometry of the irradiation field, it was within the range of the degree of uncertainty of 2.5% in the evaluation of output radiation dosage, which was described in AAPM Report 13. Therefore, the theory proposed by Khan et al. was found to be valid also for an irregular irradiation field using EDW, which could be handled in the same manner as in the irregular irradiation field without using EDW. PMID- 15753867 TI - [Method for determining scan timing based on analysis of formation process of the time-density curve]. AB - A strict determination of scan timing is needed for dynamic multi-phase scanning and 3D-CT angiography (3D-CTA) by multi-detector row CT (MDCT) . In the present study, contrast media arrival time (T(AR)) was measured in the abdominal aorta at the bifurcation of the celiac artery for confirmation of circulatory differences in patients. In addition, we analyzed the process of formation of the time density curve (TDC) and examined factors that affect the time to peak aortic enhancement (T(PA)). Mean T(AR) was 15.57+/-3.75 s. TDCs were plotted for each duration of injection. The rising portions of TDCs were superimposed on one another. TDCs with longer injection durations were piled up upon one another. Rise angle was approximately constant in response to each flow rate. Rise time (T(R)) showed a good correlation with injection duration (T(ID)). T(R) was 1.01 TID (R(2)=0.994) in the phantom study and 0.94 T(ID)-0.60 (R(2)=0.988) in the clinical study. In conclusion, for the selection of optimal scan timing it is useful to determine T(R) at a given point and to determine the time from T(AR). PMID- 15753868 TI - [Improvement in the performance of the adaptive neighborhood contrast enhancement technique based on entropy]. AB - This paper presents an improved adaptive-neighborhood-contrast-enhancement (ANCE) method for the improvement of medical image quality. The ANCE method consists of computing the local contrast around each pixel using a variable neighborhood whose size depends on the statistical properties around the given pixel. The obtained contrast image is then transformed into a new contrast image using a contrast enhancement function. Finally, a contrast-enhanced image is obtained by applying inverse contrast transform to the previous step. This technique provides the advantages of enhancing or preserving image contrast while suppressing noise. However, it does have a drawback. The performance of the ANCE method largely depends on how to determine the parameters used in the processing steps. The present study proposes a novel method for optimal and automatic determination of several parameters using entropy. To quantitatively compare the performance of the proposed method with that of the ANCE method, computer-simulated images are generated. The output-to-input SNR level and the mean squared error are used as comparison criteria. Results demonstrated the superiority of the proposed method. Moreover, we have applied our new algorithm to echocardiograms and mammograms. Our results showed that the proposed method has the potential to become useful for improving the image quality of medical images. PMID- 15753869 TI - [Fundamental study of coronary angiography using gadolinium contrast agent]. AB - To investigate the clinical usefulness of coronary arteriography using a gadolinium contrast medium (hereinafter referred to as Gd contrast medium) in patients with iodine allergy, images obtained using an iodinated contrast medium and a Gd contrast medium were compared using respective phantoms. Within the range of the effective energy (34.8 to 41.0 keV) in clinical use, the tube voltage dependency of the Gd contrast medium was lower than that of the iodinated contrast medium. The contrast ratios were found to be comparable for the images obtained by a 4-fold-diluted iodinated contrast medium and the undiluted Gd contrast medium. Although the assessment of stenotic ratio was difficult to obtain by visual evaluation of the clinical images, due to unclear borders of the blood vessels, the presence or absence of significant stenosis was discriminative by arteriography using the Gd contrast medium. Based on these findings, it was considered necessary to make a confirmed diagnosis using a Gd contrast medium with concurrent diagnosis by intravascular echography. In conclusion, coronary arteriography and PCI using a Gd contrast medium are considered to offer benefit to the diagnosis and treatment of patients with iodine allergy. PMID- 15753870 TI - [Experience in IVR performed using a posture support device designed especially for a patient unable to take the decubitus position]. AB - We were requested to perform IVR for uterine artery embolotherapy of a patient who was unable to take the decubitus position. Because of extreme overweight, the patient was limited to a sitting posture on a bed in the living environment. Therefore, a device to support her sitting posture on the IVR patient table was designed and created. The support device was used to hold the patient's body in a half-sitting posture and distribute her weight load appropriately so that the weight load applied to the patient's body could be minimized. Using this support device, IVR could be performed smoothly during a period of 2 hours 40 minutes. PMID- 15753871 TI - [Visualization of the lower cranial nerves by 3D-FIESTA]. AB - MR cisternography has been introduced for use in neuroradiology. This method is capable of visualizing tiny structures such as blood vessels and cranial nerves in the cerebrospinal fluid (CSF) space because of its superior contrast resolution. The cranial nerves and small vessels are shown as structures of low intensity surrounded by marked hyperintensity of the CSF. In the present study, we evaluated visualization of the lower cranial nerves (glossopharyngeal, vagus, and accessory) by the three-dimensional fast imaging employing steady-state acquisition (3D-FIESTA) sequence and multiplanar reformation (MPR) technique. The subjects were 8 men and 3 women, ranging in age from 21 to 76 years (average, 54 years). We examined the visualization of a total of 66 nerves in 11 subjects by 3D-FIESTA. The results were classified into four categories ranging from good visualization to non-visualization. In all cases, all glossopharyngeal and vagus nerves were identified to some extent, while accessory nerves were visualized either partially or entirely in only 16 cases. The total visualization rate was about 91%. In conclusion, 3D-FIESTA may be a useful method for visualization of the lower cranial nerves. PMID- 15753878 TI - The symptomatic accessory navicular bone: a report and discussion of the clinical presentation. AB - An accessory navicular bone is a congenital anomaly caused by aberrant ossification. Usually asymptomatic, they are relatively common in our population. They can present in several different locations, which can have an impact on the clinical presentation and the degree of dysfunction. Occasionally, these can become symptomatic, which can manifest as chronic or acute on chronic foot pain. Diagnosis of this condition relies on radiographic evaluation. The 45-degree eversion oblique view of the foot is the most important view for identifying this condition, although several different imaging techniques can be used. Treatment of this condition includes both surgical and nonsurgical options. Ultimately, surgery yields the best outcome for young patients, though conservative management has relevance for less active patients. The following case demonstrates the presence of a symptomatic accessory navicular bone in a young athlete. PMID- 15753879 TI - Color-Doppler sonography in chronic venous insufficiency: what the radiologist should know. AB - Chronic venous insufficiency (CVI) is a pathologic condition caused by valvular incompetence, with or without associated venous outflow obstruction, which may affect both the superficial and the deep venous system, causing venous hypertension and stasis. The most common form of CVI is primary varicose veins due to the insufficiency of the saphenous system. Color-Doppler sonography (CDS) is actually the main diagnostic technique of imaging for CVI. In this article, we describe the anatomy, the technique, and the information necessary to the radiologist to perform CDS in chronic venous insufficiency. The knowledge of the venous anatomy is the cornerstone for an adequate sonographic examination. The venous network in the lower extremities is divided into three systems: superficial, deep, and perforating veins. Deep veins are "comitantes" to the corresponding arteries and run under the muscular fascia. Superficial veins course into the subcutaneous fat, superficially to the deep muscular fascia; the main superficial veins are the greater and lesser saphenous and their tributaries. Connection between the saphenous veins are defined as communicating veins. Superficial and deep veins are connected by perforating veins, with flow directed, under normal circumstances, from the superficial to the deep system. The main perforating are the Hunter in the mid thigh, the Dodd in the lower thigh, the Boyd in the upper calf, and the Cockett's in the middle and lower calf. Sonographic examination must be performed in the upright and supine position. Compression sonography and color and PW Doppler are systematically employed to assess the absence of deep venous thrombosis. Femoro-popliteal veins are evaluated with color and PW Doppler for valvular insufficiency with reflux by performing Valsalva maneuver and calf compression. The sapheno-femoral and sapheno-popliteal junctions are examined to identify type of junction, continence, accessory saphenous, and incompetent collaterals. Perforating veins are usually identified at the medial aspect of the thigh and at the medial, lateral, and posterior aspects of the leg. Outward flow (lasting more than 500 ms) in the perforating veins should be considered a sign of their incompetence. Several surgical and interventional procedures are now available for the treatment of the CVI, as follows: vein ligation and stripping, stab avulsion, endoluminal occlusion of the saphenous trunks, subfascial endoscopic perforator surgery, and valvuloplasty. PMID- 15753880 TI - Diseases affecting the peribronchovascular interstitium: CT findings and pathologic correlation. AB - Central bronchi and pulmonary arteries are surrounded and enveloped by a strong connective-tissue sheath termed the peribronchovascular interstitium, extending from the level of the pulmonary hila into the peripheral lung. Thickening of the peribronchovascular interstitium can be seen in a wide variety of diseases. The CT appearance can be smooth, nodular, or irregular depending on the underlying cause. Many of the diseases affecting the peribronchovascular interstitium are entities that show a predilection for lymphatic routes, such as sarcoidosis, pulmonary lymphangitic carcinomatosis, silicosis, and lymphoproliferative disorders. There are other entities that mainly affect the peribronchovascular interstitium without a predominant perilymphatic distribution, such as hydrostatic pulmonary edema, cryptogenic organizing pneumonia, Kaposi's sarcoma, interstitial pulmonary emphysema, and interstitial hemorrhage. Although there is an overlap in radiologic features, some CT findings are useful in differentiating among these entities. When CT shows mainly peribronchovascular abnormality, the differential diagnosis is considerably reduced, and it is generally possible to reach the correct diagnosis by considering the clinical context. We illustrate the CT findings and pathologic correlation for a number of different disorders that mainly affect the peribronchovascular interstitium. PMID- 15753881 TI - Transrectal ultrasonography and ultrasound-guided biopsies of the prostate gland: how, when, and where. PMID- 15753882 TI - Targeting the IgE molecule in allergic and asthmatic diseases: review of the IgE molecule and clinical efficacy. PMID- 15753884 TI - Insect sting allergy and venom immunotherapy: a model and a mystery. AB - Whole-body extracts of Hymenoptera were used for diagnosis and treatment until controlled clinical trials proved them no better than placebo, whereas venom is 85% to 98% effective. Studies of natural history reveal why whole-body extracts were thought to work. The chance of future systemic reactions is low in large local reactors and in most children and varies between 20% and 70% in adults. Venom skin tests are most accurate, but RAST is an important complementary test. The degree of sensitivity on skin tests or RASTs does not reliably predict the severity of a sting reaction. Venom immunotherapy is recommended for patients at high risk for sting reactions. Rapid regimens are as safe as slower regimens. The recommended dose is 100 microg, but some patients require higher doses for full protection. Venom immunotherapy is continued every 4 to 8 weeks for at least 5 years in most cases. Skin test results become negative in only 25% after 5 years of therapy but in 60% to 70% after 7 to 10 years. When treatment is stopped after 5 years or more, there is a 10% chance of systemic reaction to each future sting, but most reactions are mild. Some patients have a higher risk of relapse and should continue treatment for an extended period. PMID- 15753886 TI - Molecular mechanisms of anaphylaxis: lessons from studies with murine models. AB - Studies with murine models demonstrate 2 pathways of systemic anaphylaxis: one mediated by IgE, Fc epsilonRI, mast cells, histamine, and platelet-activating factor (PAF), and the other mediated by IgG, Fc gammaRIII, macrophages, and PAF. The former pathway requires much less antibody and antigen than the latter. As a result, IgG antibody can block IgE-mediated anaphylaxis induced by small quantities of antigen without mediating Fc gammaRIII-dependent anaphylaxis. The IgE pathway is most likely responsible for most human anaphylaxis, which generally involves small amounts of antibody and antigen; similarities in the murine and human immune systems suggest that the IgG pathway might mediate disease in persons repeatedly exposed to large quantities of antigen. Mice, like human subjects, can experience IgE/Fc epsilonRI/mast cell-mediated gastrointestinal and systemic anaphylaxis in response to ingested antigen. Gastrointestinal symptoms depend on serotonin and PAF; mediator dependence of systemic symptoms has not been determined. Both local and systemic anaphylaxis induced by ingested antigens might be blocked by IgA and IgG antibodies. IL-4 and IL-13 signaling through the IL-4 receptor alpha chain, in addition to promoting the mastocytosis and IgE antibody production that mediate most human anaphylaxis, exacerbates the effector phase of anaphylaxis by increasing target cell responsiveness to vasoactive mediators. As a result, IL-4 receptor alpha chain antagonists might be particularly effective suppressors of anaphylaxis. PMID- 15753888 TI - The anti-inflammatory effects of omalizumab confirm the central role of IgE in allergic inflammation. AB - Anti-IgE therapy with omalizumab reduces serum levels of free IgE and downregulates expression of IgE receptors (Fc epsilonRI) on mast cells and basophils. In the airways of patients with mild allergic asthma, omalizumab reduces Fc epsilonRI+ and IgE+ cells and causes a profound reduction in tissue eosinophilia, together with reductions in submucosal T-cell and B-cell numbers. In patients with seasonal allergic rhinitis, omalizumab inhibits the allergen induced seasonal increases in circulating and tissue eosinophils. Omalizumab decreases Fc epsilonRI expression on circulating dendritic cells, which might lead to a reduction in allergen presentation, T(H)2 cell activation, and proliferation. As a systemic anti-IgE agent, omalizumab has demonstrated clinical efficacy in patients with moderate and severe allergic asthma and in those with seasonal and perennial allergic rhinitis, as well as in patients with concomitant allergic asthma and allergic rhinitis. The anti-inflammatory effects of omalizumab at different sites of allergic inflammation and the clinical benefits of anti-IgE therapy in patients with allergic asthma and allergic rhinitis emphasize the fundamental importance of IgE in allergic inflammation. PMID- 15753889 TI - Defining the responder in asthma therapy. PMID- 15753890 TI - Advances in pediatric and adult asthma. AB - Last year's review on adult and pediatric asthma highlighted reports related to asthma genetics, the importance of upper airway management, the costs of asthma, and the importance of early recognition and intervention. This year we will organize our discussion to review recent reports related to the origins and persistence of asthma in both adults and children. We highlight Journal publications from 2004, along with recent key publications from other medical journals, to provide a perspective on the rapidly developing areas of genetics, including pharmacogenetics, respiratory infection, biomarker measurements, and asthma pharmacotherapy. This new understanding of the pathogenesis of asthma combined with clinical applications of genetics and biomarkers should lead to new management strategies. Asthma management is likely to change in the coming years from a strategy directed to the best outcome in groups of patients to an individualized approach to assessment and management. PMID- 15753894 TI - Polymorphisms in SPINK5 are not associated with asthma in a Dutch population. AB - BACKGROUND: Asthma and allergic phenotypes are complex genetic diseases with known linkage to chromosome 5q. This region has many candidate genes, including serine protease inhibitor Kazal type 5 (SPINK5), which has been associated with asthma and atopic dermatitis in family-based studies of children with atopic dermatitis. OBJECTIVE: We sought to investigate whether single nucleotide polymorphisms in SPINK5 are associated with asthma, atopic phenotypes, and atopic dermatitis. METHODS: We investigated whether single nucleotide polymorphisms in SPINK5 (ie, -785 A/G, Asn368Ser, and Lys420Glu) are associated with asthma, atopic phenotypes, and atopic dermatitis in 200 families ascertained by a proband with asthma (nonaffected spouses served as a matched control population) and an independent set of 252 trios with asthma. RESULTS: We found no association with asthma, atopic phenotypes, and atopic dermatitis after correction for multiple testing. CONCLUSION: The negative results in this study suggest that SPINK5 is not associated with asthma or atopic phenotypes in individuals ascertained by a proband with asthma. This is consistent with the finding that SPINK5 is not expressed in the lung. Because our patients were ascertained for asthma, a role of SPINK5 in atopic dermatitis cannot be excluded. PMID- 15753892 TI - Inner City Asthma Study: relationships among sensitivity, allergen exposure, and asthma morbidity. AB - BACKGROUND: Asthma-associated morbidity is rising, especially in inner city children. OBJECTIVE: We evaluated the allergen sensitivities, allergen exposures, and associated morbidity for participants in the Inner City Asthma Study. We also determined geographic variations of indoor allergen levels. METHODS: Nine hundred thirty-seven inner city children 5 to 11 years old with moderate to severe asthma underwent allergen skin testing. Bedroom dust samples were evaluated for Der p 1, Der f 1, Bla g 1, Fel d 1, and Can f 1. RESULTS: Skin test sensitivities to cockroach (69%), dust mites (62%), and molds (50%) predominated, with marked study site-specific differences. Cockroach sensitivity was highest in the Bronx, New York, and Dallas (81.2%, 78.7%, and 78.5%, respectively), and dust mite sensitivity was highest in Dallas and Seattle (83.7% and 78.0%, respectively). A majority of homes in Chicago, New York, and the Bronx had cockroach allergen levels greater than 2 U/g, and a majority of those in Dallas and Seattle had dust mite allergen levels greater than 2 microg/g. Levels of both of these allergens were influenced by housing type. Cockroach allergen levels were highest in high rise apartments, whereas dust mite allergen levels were highest in detached homes. Children who were both sensitive and exposed to cockroach allergen had significantly more asthma symptom days, more caretaker interrupted sleep, and more school days missed than children who were not sensitive or exposed. CONCLUSION: Geographic differences in allergen exposure and sensitivity exist among inner city children. Cockroach exposure and sensitivity predominate in the Northeast, whereas dust mite exposure and sensitivity are highest in the South and Northwest. Cockroach allergen appears to have a greater effect on asthma morbidity than dust mite or pet allergen in these children. PMID- 15753895 TI - Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein. AB - BACKGROUND: Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways. The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion. In this study, we examined the hypothesis that annexin 1 surface expression, which is upregulated by the glucocorticoid receptor, prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 (cPLA2). OBJECTIVE: To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro. To determine the relationship between annexin 1surface expression and nuclear membrane translocation of cPLA2. METHODS: Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate (FP), and beta2-integrin adhesion was measured after stimulation with IL-5 or eotaxin. Effects of FP on cPLA2 expression, phosphorylation, and translocation were determined. The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides. RESULTS: Fluticasone propionate decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane. Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody. Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion. Translocation of cPLA2 to the nuclear membrane was significantly blocked by incubation with FP. Blockade was reversed with annexin 1 blocking antibody. CONCLUSION: Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1, which blocks cPLA2 translocation to nuclear membrane. PMID- 15753896 TI - Progenitor egress from the bone marrow after allergen challenge: role of stromal cell-derived factor 1alpha and eotaxin. AB - BACKGROUND: CCR3 expression on CD34+ cells mediates migration to eotaxin in vitro. CXCR4 and stromal cell-derived factor (SDF)-1alpha are important for stem cell homing to hemopoietic compartments. OBJECTIVE: To study chemokine-mediated progenitor cell traffic in allergic inflammation. METHODS: Bone marrow (BM) aspirates were obtained at baseline from normal subjects; atopic subjects without asthma; and subjects with asthma before, 5 hours after, and 24 hours after allergen inhalation (dual and early responders). Changes in chemokine receptor expression and migration were assessed. RESULTS: Expression of CXCR4, but not CCR3, on BM CD34+ cells was greater in normal subjects compared with atopic subjects with asthma. Likewise, SDF-1alpha, but not eotaxin, stimulated a greater migrational response by BM CD34+ cells from normal subjects compared with subjects with asthma. For all subjects, a positive correlation was found between intensity of CXCR4 expression and magnitude of CD34+ cell response to SDF-1alpha. Allergen inhalation attenuated both intensity of CXCR4 expression and SDF-1alpha levels in marrow from dual compared with early responders 24 hours postallergen. In contrast, the intensity of CCR3 expression on BM CD34+ cells increased in dual compared with early responders at 24 hours postallergen. In addition, an increase in migrational responsiveness of BM CD34+ cells to eotaxin and a decrease to SDF 1alpha 24 hours postallergen was found in dual responder subjects with asthma. CONCLUSION: After allergen inhalation in subjects with asthma, a downregulation in CXCR4 intensity on BM CD34+ cells and a reduction in BM SDF-1alpha levels may reduce progenitor retention to marrow stroma promoting peripheral egress, possibly mediated by the CCR3/eotaxin axis. PMID- 15753897 TI - Defective production of soluble HLA-G molecules by peripheral blood monocytes in patients with asthma. AB - BACKGROUND: HLA-G, a human nonclassic MHC class I molecule, is responsible for complex immunoinhibitory functions. HLA-G is expressed as membrane-bound and is secreted as soluble molecules by the peripheral blood CD14+ monocytes activated by IL-10. OBJECTIVE: It has been reported that LPS stimulation induces IL-10 production by PBMCs and that IL-10 levels are reduced in patients with severe asthma compared with patients with mild asthma and healthy subjects. The study was designed to investigate whether this impaired IL-10 production can affect the expression and the secretion of soluble HLA-G (sHLA-G)-1/HLA-G5 molecules. METHODS: We investigated the production of sHLA-G1/HLA-G5 and IL-10 by specific ELISAs in the culture supernatants of LPS-activated PBMCs from 24 healthy subjects and 20 patients with moderate to severe persistent asthma. RESULTS: LPS stimulation induced the secretion of IL-10 and sHLA-G1/HLA-G5 molecules in all healthy subjects, whereas in patients with asthma, the levels of IL-10 were significantly lower (P < .001) and the number of cultures exhibiting detectable sHLA-G1/HLA-G5 was reduced (7/20; P < .001). The addition of exogenous IL-10 to LPS-stimulated PBMCs from patients with asthma restored normal sHLA-G1/HLA-G5 production. CONCLUSION: Our data suggest that a specific deficit of IL-10 secretion in patients with asthma could prevent the normal production of sHLA G1/HLA-G5 molecules. The reduction of immunosuppressive activity mediated by HLA G could in turn contribute to the persistence of chronic airway inflammation in asthma. PMID- 15753899 TI - Resident CD8+ T cells suppress CD4+ T cell-dependent late allergic airway responses. AB - BACKGROUND: The role of CD8+ T cells in the immune response to airway challenge with an allergen is poorly understood. OBJECTIVE: The aim of this study was to test the hypothesis that resident naive CD8+ T cells modulate the magnitude of CD4+ T cell-dependent allergic airway responses. METHODS: Cervical lymph node CD4+ T cells (2 x 10(6)) were harvested from ovalbumin (OVA)- or sham-sensitized rats and injected intraperitoneally into naive Brown Norway recipients. The recipients were treated with a CD8alpha mAb (OX-8) to deplete the resident CD8+ T cells (n = 12) or mouse ascites (n = 12). Two days after adoptive transfer, the recipient animals were OVA challenged, lung resistance was measured for 8 hours, and bronchoalveolar lavage (BAL) was performed. RESULTS: After OVA challenge, primed CD4-transferred CD8-depleted rats had larger early airway responses and late airway responses compared with primed CD4-transferred CD8-nondepleted rats (early airway responses: 158.6% +/- 19.2% vs 115.7% +/- 5.9%, P < .05; late airway responses: 8.5% +/- 1.7% vs 4.4% +/- 0.9%, P < .05). BAL eosinophilia was also greater (4.67% +/- 0.45% vs 2.34 +/- 0.26%, P < .01). The cells in BAL fluid expressing IL-4 mRNA were not significantly changed by CD8 depletion, but IL-5 mRNA+ cells were higher in number, and IFN-gamma mRNA+ cells were fewer in the CD8-depleted group. CONCLUSIONS: Resident CD8+ T cells downregulate the late allergic response and airway inflammation evoked by CD4+ T-cell transfers in Brown Norway rats. This downregulation does not require antigen priming. PMID- 15753898 TI - Oncostatin M causes eotaxin-1 release from airway smooth muscle: synergy with IL 4 and IL-13. AB - BACKGROUND: Eotaxin is implicated in asthmatic eosinophilia. Oncostatin M (OSM) causes eotaxin release from fibroblasts. OBJECTIVE: We sought to examine the effects and mechanism of action of OSM and other IL-6 family cytokines on eotaxin release from human airway smooth muscle cells. METHODS: Eotaxin 1 release was measured by means of ELISA. Western blotting was used to examine mitogen activated protein kinase and signal transducer and activator of transcription 3 (STAT-3) phosphorylation. Eotaxin promoter activity was analyzed in cells transfected with wild-type STAT-3, a mutant form of STAT-3 that cannot be phosphorylated, and a constitutively active form of STAT-3. The mRNA and protein expression of IL-4R alpha, the signaling receptor for IL-4 and IL-13, was evaluated by means of real-time PCR and flow cytometry, respectively. RESULTS: OSM increased eotaxin 1 release and augmented IL-4- or IL-13-induced eotaxin release, whereas other IL-6 family cytokines did not. OSM caused a greater increase in STAT-3 phosphorylation and STAT-3-mediated gene transcription than other IL-6 family cytokines. OSM increased eotaxin promoter activity and augmented IL-13- and IL-4-induced increases in promoter activity. The constitutively active form of STAT-3 increased eotaxin promoter activity, whereas the mutant form of STAT-3 that cannot be phosphorylated significantly reduced eotaxin promoter activity induced by OSM or IL-4 plus OSM. OSM increased IL-4R alpha mRNA and protein levels. CONCLUSIONS: OSM induces eotaxin 1 expression in human airway smooth muscle cells by a mechanism involving STAT-3. OSM synergizes with IL-13 and IL-4 to increase eotaxin 1 expression, possibly as a result of effects on IL-4R alpha expression. PMID- 15753901 TI - Airway fibroblasts exhibit a synthetic phenotype in severe asthma. AB - BACKGROUND: Platelet-derived growth factor (PDGF) may have a significant role in airway remodeling in asthma, because it is a powerful inductor of many airway fibroblast activities such as collagen synthesis. OBJECTIVE: To determine whether PDGF is a significant contributor to airway remodeling in patients with asthma by enhancing airway fibroblast procollagen I expression. METHODS: Six normal controls without asthma, 10 subjects with mild to moderate asthma, and 5 subjects with severe asthma underwent bronchoscopy with endobronchial biopsy. Biopsies were placed in Dulbecco modified Eagle medium and fibroblasts cultured in the presence and absence of PDGF isoforms -AA, -BB, and -AB (1, 5, 10, 100 ng/mL) and insulin-like growth factor 1 (100 ng/mL). Fibroblast procollagen I and PDGF receptors (PDGFRs) alpha and beta expression were determined by ELISA. RESULTS: Platelet-derived growth factor BB significantly enhanced fibroblast procollagen I expression in patients with severe asthma compared with patients with mild/moderate asthma and normal controls. Furthermore, the baseline fibroblast expression of PDGFR-beta was significantly greater in patients with severe asthma compared with the other groups. CONCLUSION: This pilot study suggests that airway fibroblasts from patients with severe asthma exhibit a synthetic phenotype, which may be driven by the overexpression of PDGFR-beta. PMID- 15753900 TI - TGFbeta1 haplotypes and asthma in Indian populations. AB - BACKGROUND: Asthma is a complex disorder of the airways of the lungs. TGF-beta1 plays a key role in airway remodeling and asthma by having both proinflammatory and anti-inflammatory activities, making TGFbeta1 an important candidate gene to study. OBJECTIVE: To investigate the association of TGFbeta1 gene polymorphisms with asthma. METHODS: A case-control study was designed for identifying polymorphisms and haplotypes associated with asthma and associated phenotypes. We have verified our results in 2 independent cohorts collected from northern (number of patients, 187; number of controls, 187) and western India (number of patients, 209; number of controls, 190). We measured the serum TGF-beta1 levels of selected individuals and correlated them with genotypes and haplotypes. RESULTS: A novel (CT)n(CA)m repeat polymorphism (BV209662) 24.9 kb upstream of TGFbeta1 was identified. A significant association was seen at the level of alleles and genotypes with asthma in the 2 cohorts studied independently (P < .05). Interestingly, a novel 3-locus haplotype, 23_G_T, was found to be significantly associated with asthma (P = .00001 in cohorts A and B) as well as with higher serum TGF-beta1 level (P = .01). On the other hand, a novel haplotype, 22_G_C, was negatively associated with asthma (P = .00001 for cohorts A and B) and with lower serum TGF-beta1 level (P = .0019). CONCLUSION: This is the first study identifying novel risk and protective haplotypes--23_G_T and 22_G_C, respectively--in the TGFbeta1 gene that are associated with asthma. We also demonstrate the functional significance of these haplotypes with serum TGF beta1 levels. These results would be valuable in elucidating the role of TGF beta1 in asthma pathogenesis. PMID- 15753902 TI - Rhinophototherapy: a new therapeutic tool for the management of allergic rhinitis. AB - BACKGROUND: Phototherapy has a profound immunosuppressive effect and is able to inhibit hypersensibility reactions in the skin. OBJECTIVE: We evaluated whether phototherapy using a combination of UV-B (5%), UV-A (25%), and visible light (70%), referred to as mUV/VIS, is effective in treating allergic rhinitis. METHODS: We conducted a randomized, double-blind study, in 49 patients with hay fever. The study was performed during the ragweed season. Each intranasal cavity was illuminated 3 times a week for 3 weeks with mUV/VIS or with low-intensity visible light. Symptom scores, inflammatory cells, and their mediators were assessed in nasal lavages. In vitro effects of mUV/VIS irradiation on T-cell and eosinophil apoptosis and its inhibitory effect on mediator release from basophils were examined. RESULTS: Rhinophototherapy was tolerated well and resulted in a significant improvement of clinical symptoms for sneezing (P < .016), rhinorrhea (P < .007), nasal itching (P < .014), and total nasal score (P < .004). None of the scores improved significantly in the control group. Scores for nasal obstruction slightly improved after mUV/VIS treatment and significantly increased in the control group (P < .017). In the nasal lavage, phototherapy significantly reduced the number of eosinophils and the level of eosinophil cationic protein and IL-5. In vitro irradiation of T cells and eosinophils with mUV/VIS light dose dependently induced apoptosis. Furthermore, mUV/VIS irradiation inhibited the mediator release from RBL-2H3 basophils. CONCLUSION: These results suggest that phototherapy is an effective modality to treat allergic rhinitis and offer new options for the treatment of immune-mediated mucosal diseases. PMID- 15753903 TI - Association of a haplotype block spanning SDAD1 gene and CXC chemokine genes with allergic rhinitis. AB - BACKGROUND: Seasonal allergic rhinitis (SAR) is a common allergic disorder characterized by episodes of sneezing, rhinorrhea, and swelling of the nasal mucosa. Although the pathogenesis of SAR remains unclear, there does appear to be a genetic predisposition to development of SAR. We previously identified regions of chromosomes 1p, 4q, and 9q linked to SAR in 48 families (188 members) identified through children with SAR against orchard grass pollens. OBJECTIVE: The aim of the current study was to identify susceptibility genes for SAR on 4q. METHODS: We screened for markers associated with SAR on 4q with 17 microsatellite markers and then for mutations in 11 genes. We genotyped 44 single nucleotide polymorphisms (SNPs) in 48 SAR families and performed haplotype-based haplotype relative risk statistics implemented in the UNPHASED program. We also examined expression of genes with human multiple tissue and immune system cDNA panels. RESULTS: We found that 1 microsatellite marker, D4S3042, was associated with SAR (P = .034). The haplotype-based haplotype relative risk approach revealed that SNPs in SDA1 domain containing 1; chemokine, CXC motif, ligand (CXCL)-9; CXCL10; and CXCL11 were associated with SAR (P = .001-.04). These SNPs made up a haplotype block, and the most common haplotype of this block was transmitted preferentially to affected offspring (P = .002). CONCLUSION: Our results suggests that genetic variations in a haplotype block spanning the SDA1 domain containing 1 and CXC chemokine genes on 4q21 may contribute to development of SAR in the Japanese population. PMID- 15753904 TI - Analysis of the structure and allergenicity of recombinant pro- and mature Der p 1 and Der f 1: major conformational IgE epitopes blocked by prodomains. AB - BACKGROUND: The major house dust mite group 1 allergens Der p 1 and Der f 1, which belong to the papain-like cysteine protease family, are the most potent of indoor allergens. However, little information is available on the location of IgE epitopes. OBJECTIVE: We investigated the allergenicities of recombinant proforms and mature forms of Der p 1 and Der f 1 to compare them with natural Der p 1 and Der f 1 and to obtain information on the conformational IgE-binding epitopes. METHODS: Secreted pro-Der p 1 and pro-Der f 1 and their mutants without hyperglycosylation expressed in yeast were converted to mature forms. We purified the proforms and mature forms and analyzed their apparent molecular sizes and secondary structures by means of gel-filtration and circular dichroism analysis and their allergenicities by means of assays for IgE binding, IgE-binding inhibition, and basophil histamine release. The tertiary structure of pro-Der f 1 was predicted by molecular modeling. RESULTS: The recombinant mature forms exhibited similar molecular sizes, secondary structures, and allergenicities as their natural types. On the other hand, their proforms exhibited different secondary structures and less allergenicities than the mature forms in all sera and volunteers tested. Molecular modeling revealed that the prosegment is anchored at the prosegment-binding loop and the substrate-binding cleft on the surface of the mature portion. CONCLUSIONS: Our studies indicate that the prodomains of Der p 1 and Der f 1 reduce allergenicity and that the major conformational IgE epitopes commonly found in a broad population of patients exist within the 2 regions blocked by the prosegments. Recombinant Der p 1 and Der f 1 and the findings in the present study will be the basis for allergen standardization and the design of safer and more effective allergen vaccines. PMID- 15753905 TI - Relationship of validated psychometric tools to subsequent medical utilization for asthma. AB - BACKGROUND: Risk stratification is used to identify patients with asthma at increased risk of experiencing morbidity and resource utilization. Validated psychometric tools are infrequently studied sources of data for this purpose. PURPOSE: To evaluate 4 types of validated psychometric tools as predictors for subsequent asthma utilization and determine their clinical usefulness. METHODS: Eleven hundred patients with active asthma from a Health Maintenance Organization completed surveys that included demographic information and validated psychometric tools measuring generic quality of life (physical and mental components), asthma-specific quality of life, asthma control, and asthma symptom severity. Survey records were linked to administrative data that captured emergency department and hospital care, short-acting beta-agonist, and oral corticosteroid utilization for the year of and the year following the survey. Relationships of survey variables with subsequent utilization were assessed, adjusting for both baseline demographic and asthma utilization factors. RESULTS: Scores of each psychometric tool were significantly related to subsequent utilization in univariate analyses and after adjusting for baseline utilization and demographic risk factors. Patients with higher scale-defined morbidity were as much as 4 times more likely to have subsequent utilization (sensitivity as high as 58%; specificity as high as 78%). Addition of an asthma-specific tool to either demographic or utilization prediction models added sensitivity (as much as 15%) but did not substantially improve the prediction properties of models containing both demographic and utilization predictors. CONCLUSION: Validated psychometric tools appear useful for asthma risk stratification in individuals and in populations in which both utilization and demographic predictors are not available. PMID- 15753906 TI - Pathogenesis and management of anaphylaxis: current status and future challenges. PMID- 15753907 TI - Quandaries in prescribing an emergency action plan and self-injectable epinephrine for first-aid management of anaphylaxis in the community. AB - Anaphylaxis often occurs in the community in the absence of a health care professional. Prompt administration of self-injectable epinephrine as first-aid treatment in the context of a personalized emergency action plan is the key to survival. There is little argument that physicians should prescribe self injectable epinephrine for individuals who have already experienced anaphylaxis involving respiratory distress or shock triggered by allergens that might be encountered in the community. A quandary faced by physicians is that additional individuals with identified allergy who have no recognized prior history of anaphylaxis or who have a history of mild symptoms after exposure to a known trigger might also be at risk for subsequent life-threatening anaphylaxis and might also warrant prescription of self-injectable epinephrine. Prescribing for the latter individuals requires considerable clinical judgment and has led to controversy regarding possible overprescription or underprescription of self injectable epinephrine. A second quandary for physicians occurs with regard to the advice they should give to at-risk individuals about actual use of their self injectable epinephrine. It is difficult for health care professionals, let alone persons with no health care training, to predict whether anaphylaxis symptoms will occur in an at-risk individual after exposure to a known trigger. Moreover, at the onset of an acute allergic reaction, it is difficult to predict the symptoms that will ultimately develop. We examine these 2 common quandaries and provide examples of clinical scenarios and potential pitfalls in the management of persons identified as being at risk for anaphylaxis in the community. Additional studies of the recognition and treatment of anaphylaxis in the community are needed to develop comprehensive, evidence-based recommendations for its management in this setting. PMID- 15753909 TI - Statistical issues in clinical trials that involve the double-blind, placebo controlled food challenge. AB - The double-blind, placebo-controlled food challenge is a rigorous tool that has become popular for evaluating adverse reactions to foods. The standard use of the double-blind, placebo-controlled food challenge has been to document food allergies for individual patients, but it recently has been gaining acceptance as a procedure for investigating the effectiveness of therapies to prevent/minimize food-induced anaphylaxis. The purpose of this study was to describe the statistical design and analysis issues for clinical trials that use the double blind, placebo-controlled food challenge in measuring sensitivity to food allergens. Nonparametric tests for within-group and between-group comparisons are described, as well as a discrete-time survival analysis. The statistical methods are applied to simulated data from a clinical trial that compares control therapy and experimental therapy groups. The results indicate that the experimental therapy is significantly better than control in improving the tolerance to peanut flour in patients with peanut allergy. Although simple nonparametric tests for within-group and between-group comparisons are easy to apply, a discrete-time survival analysis provides the best approach because of its flexibility in accounting for important independent variables (regressors) and longitudinal data. Statistical software packages can be adapted to perform such analyses. PMID- 15753908 TI - Symposium on the definition and management of anaphylaxis: summary report. PMID- 15753910 TI - Isolated hevein-like domains, but not 31-kd endochitinases, are responsible for IgE-mediated in vitro and in vivo reactions in latex-fruit syndrome. AB - BACKGROUND: Individuals with natural rubber latex allergy often have immediate reactions to plant-derived foods and fresh fruits, such as avocado and banana. IgE of these patients has been shown to bind endochitinases containing an N terminal hevein-like domain (HLD). However, evidence on 31-kd endochitinase induced reactions in vivo is lacking. OBJECTIVE: We sought to assess the clinical significance of 31-kd endochitinases and isolated HLDs in latex-fruit syndrome. METHODS: The 31-kd endochitinases and corresponding HLDs were purified or produced from avocado, banana, latex, and wheat germ. Skin prick test reactivities against purified proteins were examined in 15 patients with natural rubber latex allergy. The binding efficiency of IgE to purified proteins was studied by using an inhibition ELISA. Experimentally resolved or modeled structures of the proteins were compared to clarify the molecular basis of clinical reactions. RESULTS: Eleven (73%) patients had skin prick test reactions to isolated HLDs of avocado and banana, but only 1 (7%) patient reacted to their corresponding 31-kd endochitinases. HLDs from avocado and banana inhibited binding of IgE to prohevein (Hev b 6.01) in 59% and 38% of patients, respectively, whereas corresponding percentages for 31-kd endochitinases were 17% and 20%, respectively. Isolated HLDs of wheat germ agglutinin and 18-kd wheat germ agglutinin did not significantly inhibit IgE binding to hevein. CONCLUSION: The isolated HLD molecules alone, but not when linked to endochitinases, seem to be responsible for IgE-mediated clinical reactions in latex-fruit syndrome. Careful selection of relevant allergens in their proper molecular form is therefore crucial in forming a reliable diagnosis of latex-fruit syndrome. PMID- 15753911 TI - Use of beta-blockers during immunotherapy for Hymenoptera venom allergy. AB - BACKGROUND: Beta-blockers may aggravate anaphylactic reactions and interfere with treatment. There is therefore concern about their use in patients who have a history of anaphylaxis or are on allergen immunotherapy. Immunotherapy is the best available treatment for prevention of life-threatening anaphylaxis to Hymenoptera stings, which is often observed in elderly patients who have cardiovascular disease and therefore are on beta-blocker treatment. OBJECTIVE: To analyze the risk of beta-blocker treatment during venom immunotherapy. METHODS: We screened all 1682 patients with Hymenoptera venom allergy seen during a period of 34 months for immunotherapy, cardiovascular disease, and treatment with beta blockers. RESULTS: Of the 1389 patients in whom immunotherapy was recommended, 11.2% had cardiovascular disease, and 44 of these were on beta-blockers before immunotherapy. In 31 of those, the drug was replaced before starting treatment. In 3 with coronary heart disease and 1 with severe ventricular arrhythmia, the drug was continued throughout immunotherapy. In 9, it was reintroduced after reaching the maintenance dose. In an additional 12 patients, beta-blockers were newly started during immunotherapy. Of 25 patients on beta-blockers during immunotherapy, 3 (12%) developed allergic side effects, compared with 23 (16.7%) of 117 with cardiovascular disease but without beta-blockers. Systemic allergic symptoms after re-exposure by sting challenge or field sting were observed in 1 of 7 (14.3%) with and 4 of 29 (13.8%) without beta-blockade. No severe reactions to treatment or sting reexposure were observed in patients with beta-blockade. CONCLUSION: Combination of beta-blockers with venom immunotherapy may be indicated in heavily exposed patients with severe cardiovascular disease. PMID- 15753912 TI - Sol i 1, the phospholipase allergen of imported fire ant venom. AB - BACKGROUND: Sol i 1, the venom phospholipase of imported fire ant venom is an important allergen and exhibits some cross-reactivity with IgE antibodies from patients sensitized to other Hymenoptera venoms. OBJECTIVE: To determine the primary structure of Sol i 1 and evaluate the roles of protein and carbohydrate epitopes in its cross-reactivity. METHODS: Sol i 1 was purified from venom, proteolytic peptides prepared and amino acid sequences obtained. The cDNA for Sol i 1 was cloned, sequenced, and compared with sequences of other wasp venom phospholipases. The role of carbohydrate epitopes in the cross-reactivity with other Hymenoptera venoms was studied by RAST inhibition. RESULTS: The sequence identified Sol i 1 as a lipase of the GX class, lipoprotein lipase superfamily, pancreatic lipase homologous family and RP2 subgroup phospholipases as are the vespid venom phospholipases. The 148 residues identified by amino acid sequencing represent about 48% of the translated cDNA sequence. Sol i 1 was 31-32% identical to yellow jacket phospholipases. The identical regions of sequence were clustered in the domain which forms the serine hydrolase active site. Mannosylated N glycans could completely inhibit binding of IgE from honeybee venom sensitized patients to Sol i 1. Inhibition by glycan of IgE binding from yellow jacket venom sensitized patients was low or absent for three of eight sera and substantial, but not complete for five sera. CONCLUSIONS: Sol i 1 is related to wasp venom phospholipases. Cross-reactivity with honeybee venom is caused by carbohydrate, whereas cross-reactivity with yellow jacket venom involves reactivity with both carbohydrate determinants of hyaluronidase and high molecular weight proteins and phospholipase protein determinants. PMID- 15753913 TI - IgE-mediated anaphylaxis caused by bites of the pigeon tick Argas reflexus: cloning and expression of the major allergen Arg r 1. AB - BACKGROUND: Anaphylactic reactions caused by bites of the European pigeon tick Argas reflexus are repeatedly reported. This soft-backed tick is a parasite of wild pigeons colonizing urban buildings and houses. Occasionally the ticks can bite human beings, inducing anaphylactic reactions in sensitized patients. OBJECTIVE: Our aim was to characterize the major allergen implicated in a series of anaphylactic reactions caused by Argas bites and to produce the allergen as recombinant protein for diagnostic purposes. METHODS: Protein extracts were prepared from whole A reflexus bodies, and IgE immunoblots were performed with sera from 13 patients who had an anaphylactic reaction with pigeon tick bites. A cDNA expression library was constructed from whole ticks and screened with a polyclonal rabbit antiserum raised against the major allergen. RESULTS: The cDNA coding for the dominant allergen Arg r 1 could be isolated. It encodes a protein belonging to the lipocalin family. Allergenicity of the recombinant Arg r 1 was confirmed by immunoblot, ELISA, and intradermal skin tests. CONCLUSION: The dominant allergen of A reflexus has been isolated and the corresponding cDNA cloned. The recombinant protein, a lipocalin, was expressed in Escherichia coli and was shown to be immunoreactive in vitro and in vivo. Recombinant Arg r 1 was used as a diagnostic tool in a series of anaphylactic reactions caused by pigeon tick bites. PMID- 15753916 TI - Banana anaphylaxis with a negative commercial skin test. PMID- 15753915 TI - Toll-like receptor 2-positive and Toll-like receptor 4-positive cells in adenoids of children exposed to passive smoking. PMID- 15753914 TI - Proteinase-activated receptor 2 activation in the airways enhances antigen mediated airway inflammation and airway hyperresponsiveness through different pathways. AB - BACKGROUND: Serine proteinases such as mast cell tryptase, trypsin-like enzymes, and certain allergens are important in the pathogenesis of asthma. These proteinases can activate the proteinase-activated receptor (PAR)-2, which has been shown to be upregulated in the airways of patients with asthma. OBJECTIVE: The purpose of this study was to investigate PAR-2 activation in the airways during allergen challenge and its effects on the 2 principle features of asthma, airway inflammation and airway hyperresponsiveness (AHR). METHODS: Proteinase activated receptor 2 activating peptide SLIGRL-NH2 (PAR-2 activating peptide [ap]) or control peptide LSIGRL-NH2 (PAR-2 control peptide [cp]) was administered alone or in conjunction with ovalbumin intranasally to mice, and AHR and airway inflammation were evaluated. RESULTS: PAR2ap did not induce AHR or airway inflammation in ovalbumin-sensitized mice that had not been challenged with ovalbumin. When administered with ovalbumin, PAR-2ap enhanced AHR and airway inflammation compared with ovalbumin administered alone or with PAR-2cp. The enhanced AHR persisted for 5 days, whereas the enhancement to airway inflammation dissipated. Mice administered PAR-2ap alone during the 5 days after the final antigen challenge demonstrated an additional enhancement to airway inflammation compared with the control animals. PAR-2ap administered with allergen increased TNF and IL-5 mRNA in lung tissue and IL-13 and TNF in bronchoalveolar lavage fluid. CONCLUSION: Exogenous PAR-2 activation in parallel with allergen challenge enhances allergen-mediated AHR and airway inflammation through distinct mechanisms. PAR-2 activation can also enhance established airway inflammation even when dissociated from exposure to allergen. Therefore, PAR-2 activation may play a pathogenic role in the development of AHR and airway inflammation. PMID- 15753917 TI - Anaphylactic shock caused by a selective allergy to celecoxib, with no allergy to rofecoxib or sulfamethoxazole. PMID- 15753918 TI - IgE-mediated asthma associated with a unique allergen from Angelim pedra (Hymenolobium petraeum) wood. PMID- 15753919 TI - SPINK5 polymorphism is associated with disease severity and food allergy in children with atopic dermatitis. PMID- 15753920 TI - Hycor manufactures truly quantitative allergen-specific IgE assays. PMID- 15753921 TI - Apparent lack of response to epinephrine in anaphylaxis. PMID- 15753922 TI - The Ishizakas and the search for reaginic antibodies. PMID- 15753923 TI - The story of IgND. PMID- 15753924 TI - Science behind the discovery of IgE. PMID- 15753926 TI - The diagnosis and management of anaphylaxis: an updated practice parameter. PMID- 15753927 TI - A dwell position verification method for high dose rate brachytherapy. AB - Misplacement of dwell positions is a potential source of misadministration in high dose rate brachytherapy. In this work we present a dwell position verification method using fluoroscopic images. A mobile C-arm fluoroscopic machine is used to take a snapshot of the treatment machine's check cable as it reaches the most distal dwell position. This fluoroscopic image is displayed side by-side with a treatment planning image on a dual monitor relay station at the HDR treatment console. Any discrepancy between the check cable's position on the verification image and the intended dwell position on the planning image can be identified immediately, thus avoiding the possibility of treating the wrong target volume. PMID- 15753929 TI - Quality assurance devices for dynamic conformal radiotherapy. AB - Two quality control devices, light-field device and radiation-field device, have been specially designed to facilitate the clinical implementation of conformal dynamic arc treatment (CDAT) and intensity modulated radiation therapy (IMRT). With the devices, the light field as well as the radiation field, projected from the individual beam at any treatment position (i.e. arbitrary gantry angle) can be evaluated. For application, the device was attached at the front end of the couch and was placed at the isocentre of the linear accelerator treatment system (LINAC). The devices are designed to be rotated parallel to the gantry head so that the light field and the radiation field projected from a direct beam can be assessed. The aim of the study is, with the aid of the devices, to evaluate the geometric precision of the beam placement and the dosimetric accuracy performed in CDAT and IMRT. The devices are placed separately from the LINAC during application and provide an independent check on the quality performance of the LINAC in three dimensions. The condition of gantry sag and any mechanical displacement resulting in field shift can be observed and traced during gantry rotation. Mistakes that occurred during the isocentre calibration can lead to significant displacement in the field projection, which would not be revealed with the conventional quality control setting (i.e. gantry 0(o)), was demonstrated with the aid of the devices here. The influence of gravitational acceleration in MLC leaf positioning error, which would consequently lead to inaccurate dose delivery, was investigated. The results of our study show that the existence of gravitational influence is statistically significant, although the magnitude of the dose inaccuracy has been found to be small. PMID- 15753930 TI - Dosimetric benefits of respiratory gating: a preliminary study. AB - In this study, we compared the amount of lung tissue irradiated when respiratory gating was imposed during expiration with the amount of lung tissue irradiated when gating was imposed during inspiration. Our hypothesis was that the amount of lung tissue spared increased as inspiration increased. Computed tomography (CT) image data sets were acquired for 10 patients who had been diagnosed with primary bronchogenic carcinoma. Data sets were acquired during free breathing and during breath-holds at 0% tidal volume and 100% tidal volume, and, when possible, at deep inspiration, corresponding to approximately 60% vital capacity. Two treatment plans were developed on the basis of each of the gated data sets: one in which the treatment portals were those of the free-breathing plan, and the other in which the treatment portals were based on the gated planning target volumes. Dose-mass histograms of the lungs calculated at 0% tidal volume were compared to those calculated at deep inspiration and at 100% tidal volume. Data extracted from the dose-mass histograms were used to determine the most dosimetrically beneficial point to gate, the reduction in the amount of irradiated lung tissue that resulted from gating, and any disease characteristics that might predict a greater need for gating. The data showed a reduction in the mass of normal tissue irradiated when treatment portals based on the gated planning target volume were used. More normal lung tissue was spared at deep inspiration than at the other two gating points for all patients, but normal lung tissue was spared at every point in the respiratory cycle. No significant differences in the amount of irradiated tissue by disease characteristic were identified. Respiratory gating of thoracic radiation treatments can often improve the quality of the treatment plan, but it may not be possible to determine which patients may benefit from gating prior to performing the actual treatment planning. PMID- 15753932 TI - Anthropomorphic breast phantoms for quality assurance and dose verification. AB - An evaluation of two anthropomorphic breast phantoms, which have been designed for quality assurance and dose verification of radiotherapy treatment of breast cancer patients, is presented. These phantoms are identical in terms of their dimensions and shape, and composed of several layers of either Plastic Water or tissue-equivalent material. Both water- and tissue-equivalent phantoms include lung- and rib-equivalent components. The phantoms simulate large, medium and small breasts. The value of the phantoms as breast treatment quality assurance tools was assessed by dose measurements with ionization chamber and thermoluminescence dosimeters (TLD), at different points inside the phantom. Measurements were made by irradiating the phantoms under conditions representing the different treatment techniques, found by the Radiological Physics Center (RPC) during its dosimetry quality audits. Most irradiations were performed with the water-equivalent breast phantom. One experiment was performed under consistent irradiation conditions to compare the tissue-equivalent phantom with the water-equivalent phantom. Measurements were compared with the dose estimated by the RPC's manual calculations used to check clinical charts of patients entered in a National Surgical Adjuvant Breast and Bowel Project (NSABP) protocol. Measurements were also compared with isodose distributions generated by a commercial radiation treatment planning (RTP) system. In the homogeneous three dimensional (3-D) phantom, fairly good agreement (within 5%) was observed at the NSABP dose prescription point between measurements and 2-D dose estimation by manual calculations. At the same dose prescription point, but located in the heterogeneous 3-D phantom, agreement between measurements and a 3-D RTP system was within about 3%. Manual calculation resulted in overestimation of up to 6%. The general agreement between the TLD measurements and the 2-D RTP values was within 3% at various off-axis points, with the exception of a few points far off axis, near the high-dose gradient region at the surface of the phantom. PMID- 15753931 TI - Quality assurance of registration of CT and MRI data sets for treatment planning of radiotherapy for head and neck cancers. AB - We are implementing the use of magnetic resonance (MR) images for head and neck radiotherapy planning, which involves their registration with computed tomography (CT). The quality assurance (QA) of the registration process was an initial step of this program. A phantom was built, and appropriate materials were identified to produce clinically relevant MR T1 and T2 contrast for its constituent "anatomy". We performed a characterization of the distortion detectable within our phantom. Finally, we assessed the accuracy of image registration by contouring structures in the registered/fused data sets using the treatment planning system. Each structure was contoured using each modality, in turn, blind of the other. The position, area, and perimeter of each structure were assessed as a measure of accuracy of the entire image registration process. Distortion effects in the MR image were shown to be minimized by choosing a suitable (3 +/- 30 kHz) receiver bandwidth. Remaining distortion was deemed clinically acceptable within +/-15 cm of the magnetic field isocenter. A coefficient of agreement (A) analysis gave values to be within 9% of unity, where A = square root(RaRp) and Ra/p is the ratio of the area/perimeter of a particular structure on CT to that on MR. The center of each structure of interest agreed to within 1.8 mm. A QA process has been developed to assess the accuracy of using multimodality image registration in the planning of radiotherapy for the head and neck; we believe its introduction is feasible and safe. PMID- 15753933 TI - A TCP-NTCP estimation module using DVHs and known radiobiological models and parameter sets. AB - Radiotherapy treatment plan evaluation relies on an implicit estimation of the tumor control probability (TCP) and normal tissue complication probability (NTCP) arising from a given dose distribution. A potential application of radiobiological modeling to radiotherapy is the ranking of treatment plans via a more explicit determination of TCP and NTCP values. Although the limited predictive capabilities of current radiobiological models prevent their use as a primary evaluative tool, radiobiological modeling predictions may still be a valuable complement to clinical experience. A convenient computational module has been developed for estimating the TCP and the NTCP arising from a dose distribution calculated by a treatment planning system, and characterized by differential (frequency) dose-volume histograms (DDVHs). The radiobiological models included in the module are sigmoidal dose response and Critical Volume NTCP models, a Poisson TCP model, and a TCP model incorporating radiobiological parameters describing linear-quadratic cell kill and repopulation. A number of sets of parameter values for the different models have been gathered in databases. The estimated parameters characterize the radiation response of several different normal tissues and tumor types. The system also allows input and storage of parameters by the user, which is particularly useful because of the rapidly increasing number of parameter estimates available in the literature. Potential applications of the system include the following: comparing radiobiological predictions of outcome for different treatment plans or types of treatment; comparing the number of observed outcomes for a cohort of patient DVHs to the predicted number of outcomes based on different models/parameter sets; and testing of the sensitivity of model predictions to uncertainties in the parameter values. The module thus helps to amalgamate and make more accessible current radiobiological modeling knowledge, and may serve as a useful aid in the prospective and retrospective analysis of radiotherapy treatment plans. PMID- 15753934 TI - Results of a one year survey of output for linear accelerators using IMRT and non IMRT techniques. AB - This paper presents the results of a one year survey of treated fields for 3 treatment machines at our New Jersey regional center. One machine predominantly treated IMRT prostate patients using a sliding window technique. The others were not equipped to deliver IMRT. Information obtained for each treated field included patient number, modality, monitor units delivered, gantry angle, and time. Data was obtained directly from our record and verify system and analyzed using a spreadsheet. We studied workload (MU/week), patient load, and average MU per patient as a function of time, as well as angular distributions and number of treatment fractions per patient. We also calculated the fraction of time the beam was on during treatments. By the end of the survey year, the workload of the IMRT machine reached approximately 100,000 MU/week and that of the non-IMRT machines was approximately 40-45000 MU/week. This was due predominantly to the higher number of monitor units for IMRT plans. Patient loads were not significantly different for the 3 machines. Duty cycle was 14% and 16% for the non-IMRT machines and 27% for the IMRT machine. The difference in workload for IMRT treatments relative to non-IMRT treatments confirms an earlier study performed at our institution using a much smaller data sample. One needs to consider the increase in leakage associated with this higher workload when designing shielding for an IMRT room. PMID- 15753935 TI - Commissioning and implementation of a stereotactic conformal radiotherapy technique using a general-purpose planning system. AB - The purpose of this paper is to report on commissioning and clinical implementation of a customized system for pediatric stereotactic conformal radiotherapy (SCRT). The system is based on the Pinnacle treatment-planning system and its interfaces with other equipment: (1) Beam models were optimized for our compact blocking system and a new LINAC. (2) Three CT-to-density conversion tables were evaluated, one using tabulated data for a commercial phantom, the second including additional points from the manufacturer's data for the inserts in an in-house phantom, and the third using measured densities for the in-house phantom materials combined with tabulated data for the commercial phantom. (3) Blocks were transferred to a computerized block cutter using in house software that extracted the block shape from the export file and custom fitted the additional necessary shapes. (4) In the absence of a DICOM RT Image link, a method based on screen data capture was used to export digitally reconstructed radiographs (DRRs) to two portal imaging systems for treatment verification. Lens shielding by multileaf collimation in the anterior-posterior isocenter verification field was investigated. (1) Computed dose distributions using the beam models agreed with measurements well within published acceptability criteria. A difference of up to 1.0 mm was measured between the beam's eye views of aperture blocks and computed 50% isodose contours for a 2 x 2 x 2 mm dose calculation grid. (2) The third table, which included measured densities, improved the accuracy of the calculated isocenter dose by up to 0.5% in typical patient SCRT treatments and up to 1.0% in a phantom with 5-cm diameter inhomogeneity inserts. (3) The block export and customization process was shown to introduce no additional uncertainty. A 1-mm block production uncertainty was measured using film dosimetry on six blocks. (4) The DRR transfer method did not introduce uncertainty into the process. Verification field shielding reduced lens dose by 12 to 15 times. In conclusion, this customized system for planning and verification of pediatric SCRT provides a high level of precision as well as reasonable practical efficiency. PMID- 15753937 TI - Compensators: an alternative IMRT delivery technique. AB - Seven years of experience in compensator intensity-modulated radiotherapy (IMRT) clinical implementation are presented. An inverse planning dose optimization algorithm was used to generate intensity modulation maps, which were delivered via either the compensator or segmental multileaf collimator (MLC) IMRT techniques. The in-house developed compensator-IMRT technique is presented with the focus on several design issues. The dosimetry of the delivery techniques was analyzed for several clinical cases. The treatment time for both delivery techniques on Siemens accelerators was retrospectively analyzed based on the electronic treatment record in LANTIS for 95 patients. We found that the compensator technique consistently took noticeably less time for treatment of equal numbers of fields compared to the segmental technique. The typical time needed to fabricate a compensator was 13 min, 3 min of which was manual processing. More than 80% of the approximately 700 compensators evaluated had a maximum deviation of less than 5% from the calculation in intensity profile. Seventy-two percent of the patient treatment dosimetry measurements for 340 patients have an error of no more than 5%. The pros and cons of different IMRT compensator materials are also discussed. Our experience shows that the compensator-IMRT technique offers robustness, excellent intensity modulation resolution, high treatment delivery efficiency, simple fabrication and quality assurance (QA) procedures, and the flexibility to be used in any teletherapy unit. PMID- 15753938 TI - Commissioning and quality assurance for intensity modulated radiotherapy with dynamic multileaf collimator: experience of the Pontificia Universidad Catolica de Chile. AB - The objective of this paper is to present our experience in the commissioning and quality assurance (QA) for intensity modulated radiotherapy (IMRT) using dynamic multileaf collimator (dMLC), sliding window technique. Using Varian equipment solution, the connectivity and operation between all IMRT chain components was checked. Then the following test were done: stability of leaf positioning and leaf speed, sensitivity to treatment interruptions (acceleration and deceleration), evaluation of standard field patterns, stability of dMLC output, segmental dose accuracy check, average leaf transmission, dosimetric leaf separation, effects of lateral disequilibrium between adjacent leaves in dose profiles and multiple carriage field verification. Standard patterns were generated for verification: uniform field, pyramid, hole, wedge, peaks and chair. Weekly QA Protocol include: sweeping gap output, Garden Fence Test (narrow bands, 2 mm wide, of exposure spaced at 2-cm intervals) and segmental dose accuracy check. Monthly QA include: sweeping gap output at multiple gantry and collimator angle, sweeping gap output off-axis, Picket Fence Test (eight consecutive movements of a 5-cm wide rectangular field spaced at 5-cm intervals), stability of leaf speed and leaf motor current test (PWM test). Patient QA procedure consists of an absolute dose measurement for all treatments fields in the treatment condition, analysis of actual leaf position versus planned leaf position (dynalog files) for each treatment field, film relative dose determination for each field, film relative dose determination for the plan (all treatment fields) in two axial planes and patient positioning verification with orthogonal films. The tests performed showed acceptable result. After more than one year of IMRT treatment the routine QA machine checks confirm the precision and stability of the IMRT system. PMID- 15753939 TI - Quality assurance evaluation of delivery of respiratory-gated treatments. AB - We describe a method for evaluating the quality of respiratory-gated radiation delivery using a commercially available device. During irradiation, gating traces for one field for each treatment were extracted from the system for each of 14 patients. The data were then transferred to a spreadsheet. Software was developed to evaluate the following parameters: duty cycle, amplitude of fiducial motion, fraction of amplitude of motion during gated delivery, and respiratory cycle time. Criteria were established for acceptability of gating traces. In our sample, over 85% of the traces indicated acceptability. An example of results for one patient extracted from analyzed gating traces is as follows: mean duty cycle, 57%, average amplitude of motion, 0.89 cm, average fraction of motion during gated delivery, 0.45; mean respiratory cycle time, 4.5 s. This technique can be used to evaluate delivery of respiratory-gated radiation therapy for quality assurance purposes and to assess various techniques for improving delivery of gated therapy. A hardcopy of the gating traces can be used to document gated treatment delivery for potential billing of the gated delivery process. PMID- 15753940 TI - Improving a scissor-action couch for conformal arc radiotherapy and radiosurgery. AB - We have developed a method to improve the setup accuracy of a Varian Clinac 6/100 couch for delivering conformal arc therapy using a tertiary micro multileaf collimator (MLC) system. Several immobilization devices have been developed to improve the mechanical stability and isocenter alignment of the couch: turn-knob harnesses, double-track alignment plates, and a drop-in rod that attaches the couch to the concrete floor. These add-on components minimize the intercomponent motion of the couch's scissor elevator, which allows consistent treatment setup. The accuracy of our isocenter couch alignment is an improvement over the above devices, within 1 mm of their accuracy. The couch has been used with over 15 patients and with over 50 modulated conformal arc treatment deliveries at our institution. PMID- 15753941 TI - Clinical validation of the normalized mutual information method for registration of CT and MR images in radiotherapy of brain tumors. AB - Image registration integrates information of different imaging modalities and has the potential to improve target volume determination in radiotherapy planning. This paper describes the implementation and validation of a 3D fully automated registration procedure in the process of radiotherapy treatment planning of brain tumors. 15 Patients with various brain tumors received CT and MR brain imaging before the start of radiotherapy. The normalized mutual information (NMI) method was used for image registration. Registration accuracy was estimated by performing statistical analysis of coordinate differences between CT and MR anatomical landmarks along the x-, y- and z-axes. Second, a visual validation protocol was developed to validate the quality of individual registration solutions and this protocol was tested in a series of 36 CT-MR registration procedures with intentionally applied registration errors. The mean coordinate differences between CT and MR landmarks along the x- and y-axes were in general within 0.5 mm. The mean coordinate differences along the z-axis were within 1.0 mm, which is of the same magnitude as the applied slice thickness in scanning. Second, the detection of intentionally applied registration errors by employment of a standardized visual validation protocol resulted in low false-negative and low false-positive rates. Application of the NMI method for the brain results in excellent automatic registration accuracy and the method has been incorporated in daily routine within our institute. A standardized validation protocol is proposed that ensures the quality of individual registrations by detecting registration errors with high sensitivity and specificity. This protocol is proposed for the validation of other linear registration methods. PMID- 15753942 TI - Linac-based stereotactic radiosurgery for treatment of trigeminal neuralgia. AB - Trigeminal neuralgia (TN) is a disabling pain condition that has classically been treated using either surgical or medical techniques. Several researchers have shown that stereotactically delivered radiation can be an effective tool in the amelioration of this condition. For these studies, the Gamma Knife was used to deliver the radiation treatment. The target location was designated as the proximal nerve at the root entry zone, and doses greater than 70 Gy to the maximum point in a single fraction were found to be effective in controlling pain in 80% of the patients treated. LINAC-based stereotactic radiosurgery has been notably absent from the treatment of TN, even though it has many similarities to Gamma Knife-based stereotactic radiosurgery. The aim of this paper is to describe our LINAC-based stereotactic technique for treatment of TN. We also compare treatment of TN using our technique to that using the Gamma Knife. We found that a LINAC-based treatment of TN can be accomplished with accuracy comparable to treatments delivered using the Gamma Knife. The dose distributions are essentially equivalent for the two treatment approaches. The LINAC-based system is easy to plan and offers the ability to reduce the involvement of sensitive structures from the treatment fields as well as the Gamma Knife system does. A disadvantage of the LINAC-based system is the time involved for treatment. PMID- 15753944 TI - Hydrogen sulphide is a mediator of carrageenan-induced hindpaw oedema in the rat. AB - Hydrogen sulphide (H(2)S) is a naturally occurring gas, with potent vasodilator activity. In this report, we identify a role for H(2)S in carrageenan-induced hindpaw oedema in the rat. Intraplantar injection of carrageenan (150 microl, 2% (w v(-1))) resulted in an increase in hindpaw H(2)S synthesising enzyme activity and increased myeloperoxidase (MPO) activity. Pretreatment (i.p. 60 min before carrageenan) with DL-propargylglycine (PAG, 25-75 mg kg(-1)), an inhibitor of the H(2)S synthesising enzyme cystathionine-gamma-lyase (CSE), significantly reduced carrageenan-induced hindpaw oedema in a dose-dependent manner (e.g. increase in hindpaw weight at 3 h, saline: 0.12+/-0.017 g; carrageenan, 1.39+/-0.037 g; PAG, 50 mg kg(-1), 1.11+/-0.06 g, n=10) and MPO activity (fold increase) in the hindpaw (saline: 1.0+/-0.12; carrageenan, 2.92+/-0.45 g; PAG, 50 mg kg(-1), 1.1+/ 0.22, n=10); PAG (50 mg kg(-1)) also inhibited H(2)S synthesising enzyme activity (nmol microg DNA(-1)) in the hindpaw in a dose-dependent manner (saline, 0.46+/ 0.05; carrageenan, 0.71+/-0.08 g; PAG, 50 mg kg(-1), 0.17+/-0.05, n=10). PMID- 15753945 TI - Curcumin modulation of Na,K-ATPase: phosphoenzyme accumulation, decreased K+ occlusion, and inhibition of hydrolytic activity. AB - 1 Curcumin, the major constitute of tumeric, is an important nutraceutical that has been shown to be useful in the treatment of many diseases. As an inhibitor of the sarcoplasmic reticulum Ca(2+)-ATPase, curcumin was shown to correct cystic fibrosis (CF) defects in some model systems, whereas others have reported no or little effects on CF after curcumin treatment, suggesting that curcumin effect is not due to simple inhibition of the Ca(2+)-ATPase. 2 We tested the hypothesis that curcumin may modulate other members of the P(2)-type ATPase superfamily by studying the effects of curcumin on the activity and kinetic properties of the Na,K-ATPase. 3 Curcumin treatment inhibited Na,K-ATPase activity in a dose dependent manner (K(0.5) approximately 14.6 microM). Curcumin decreased the apparent affinity of Na,K-ATPase for K(+) and increased it for Na(+) and ATP. Kinetic analyses indicated that curcumin induces a three-fold reduction in the rate of E1P --> E2P transition, thereby increasing the steady-state phosphoenzyme level. Curcumin treatment significantly abrogated K(+) occlusion to the enzyme as evidenced from kinetic and proteolytic cleavage experiments. Curcumin also significantly decreased the vanadate sensitivity of the enzyme. 4 Thus, curcumin partially blocks the K(+) occlusion site, and induces a constitutive shift in the conformational equilibrium of the enzyme, towards the E1 conformation. 5 The physiological consequences of curcumin treatment previously reported in different epithelial model systems may, at least in part, be related to the direct effects of curcumin on Na,K-ATPase activity. PMID- 15753946 TI - Muscarinic receptor binding, plasma concentration and inhibition of salivation after oral administration of a novel antimuscarinic agent, solifenacin succinate in mice. AB - 1 A novel muscarinic receptor antagonist, solifenacin succinate, inhibited specific binding of [N-methyl-(3)H]-scopolamine ([(3)H]-NMS) in the mouse bladder, submaxillary gland and heart in a concentration-dependent manner. This inhibitory effect was greatest in the submaxillary gland, followed by the bladder and heart. 2 After oral administration of oxybutynin (76.1 micromol kg(-1)) or solifenacin (62.4, 208 micromol kg(-1)), a significant dose- and time-dependent increase in K(D) values for specific [(3)H]-NMS binding was seen in the bladder, prostate, submaxillary gland, heart, colon and lung, compared with control values. The increase in K(D) induced by oxybutynin in each tissue reached a maximum 0.5 h after oral administration and then rapidly declined, while that induced by solifenacin was greatest 2 h after administration and was maintained for at least 6 or 12 h, depending on the dose. The muscarinic receptor binding of oral solifenacin was slower in onset and of a longer duration than that of oxybutynin. 3 Plasma concentrations of oxybutynin and its active metabolite (N desethyl-oxybutynin, DEOB) were maximum 0.5 h after its oral administration and then declined rapidly. Oral solifenacin persisted in the blood for longer than oxybutynin. 4 Pilocarpine-induced salivary secretion in mice was significantly reduced by oral administration of solifenacin and was completely abolished 0.5 h after oral oxybutynin. Although the suppression induced by solifenacin was more persistent than that due to oxybutynin, the antagonistic effect of solifenacin on the dose-response curves to pilocarpine was significantly weaker than that of oxybutynin. It is concluded that oral solifenacin persistently binds to muscarinic receptors in tissues expressing the M(3) subtype, such as the bladder. PMID- 15753947 TI - Design and characterization of a new cell-permeant inhibitor of the beta secretase BACE1. AB - 1 The beta-secretase BACE1 is one of the enzymes that contribute to the production of the Abeta peptide, in vitro and in vivo. JMV1195 was previously shown to inhibit BACE activity in vitro but was unable to block cellular BACE activity. We have designed a new permeable inhibitor, JMV2764 that corresponds to a derivative of JMV1195 to which a penetratin sequence had been added at its N terminus. We have assessed the ability of JMV2764 to affect BACE1 activity in vitro, and to modify Abeta production in various cell systems. 2 Endogenous beta secretase or BACE1 activities were monitored in vitro by means of two distinct fluorimetric substrates in HEK293 extracts of cells expressing either wild-type betaAPP, Swedish mutated betaAPP or SPA4CT constructs. Abeta40 recovery was monitored by immunoprecipitation and Western blot analysis. 3 JMV2764 and JMV1195 inhibited endogenous beta-secretase activity of HEK293 cellular homogenates with IC(50)s of 0.8 and 6.6 microM, respectively. Interestingly, JMV2764 also inhibited beta-secretase activity after preincubation with intact cells while JMV1195 was inactive, indicating that unlike JMV1195, JMV2764 could penetrate into the cells. 4 JMV2764 but not JMV1195 also prevented Abeta production by HEK293 cells overexpressing wild-type and Swedish-mutated betaAPP. However, JMV2764 was unable to affect Abeta production from cells expressing SPA4CT, a betaAPP-derived sequence that does not need beta-secretase to produce Abeta. 5 Altogether, we have designed a new cell-permeable BACE1 inhibitor that allows to envision to prevent Abeta production in vivo. Work is in progress to assess the potential of these compounds to prevent Abeta production in transgenic mice overproducing Abeta. PMID- 15753948 TI - Human breast cancer cell line MDA-MB-231 expresses endogenous A2B adenosine receptors mediating a Ca2+ signal. AB - 1 Two human breast cancer cell lines, MCF-7 and MDA-MB-231, were screened for the presence of functionally significant adenosine receptor subtypes. 2 MCF-7 cells did not contain adenosine receptors as judged by the lack of an effect of nonselective agonists on adenylyl cyclase activity or intracellular Ca(2+) levels. MDA-MB-231 cells showed both a stimulation of adenylyl cyclase and a PLC dependent increase in intracellular Ca(2+) in response to nonselective adenosine receptor agonists. 3 Both adenosine-mediated responses in MDA-MB-231 cells were observed with the nonselective agonists 5'-N-ethylcarboxamidoadenosine (NECA) and 2-(3-hydroxy-3-phenyl)propyn-1-yladenosine-5'-N-ethyluronamide (PHPNECA), but no responses were observed with agonists selective for A(1), A(2A) or A(3) adenosine receptors. The Ca(2+) signal was antagonized by 8-cyclopentyl-1,3 dipropylxanthine (DPCPX) and the nonselective antagonist 9-ethyl-8-furyladenine (ANR 152), but not by A(2A) or A(3) selective compounds. 4 In radioligand binding with [2-(3)H](4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5 ylamino]ethyl)phenol) ([(3)H]ZM 241385), a specific binding site with a K(D) value of 87 nM and a B(max) value of 1600 fmol mg(-1) membrane protein was identified in membranes from MDA-MB-231 cells. 5 The pharmacological characteristics provide evidence for the expression of an A(2B) adenosine receptor in MDA-MB-231 cells, which not only mediates a stimulation of adenylyl cyclase but also couples to a PLC-dependent Ca(2+) signal, most likely via G(q/11). The A(2B) receptor in such cancer cells may serve as a target to control cell growth and proliferation. 6 The selective expression of high levels of endogenous A(2B) receptors coupled to two signaling pathways make MDA-MB-231 cells a suitable model for this human adenosine receptor subtype. PMID- 15753950 TI - The role of nitric oxide synthase in reduced vasocontractile responsiveness induced by prolonged alpha1-adrenergic receptor stimulation in rat thoracic aorta. AB - 1 Prolonged exposure (6-12 h) of rat aorta to alpha1-adrenergic receptor (alpha(1)AR) agonist phenylephrine (Phe) leads to a decrease in alpha(1)AR mediated vasoconstriction. This reduced responsiveness to alpha(1)AR stimulation was strongly dependent on the intactness of the endothelium. 2 We examined the effect of Phe on nitric oxide synthase (NOS) activity by measuring the conversion of [(3)H]L-arginine to [(3)H]L-citrulline in rat aorta or in endothelial cells isolated from rat aorta. Phe stimulation increased NOS activity in control aortas. This response was antagonized by prazosin. However, Phe increased neither the activity of NOS nor intracellular Ca(2+) in the isolated endothelial cells from the control aortas, whereas acetylcholine (Ach) was able to stimulate both responses in these cells. This result suggests that Phe stimulates alpha(1)AR on vascular smooth muscle cells and has an indirect influence on endothelial cells to increase NOS activity. 3 In Phe-exposed aortic rings, basal NOS activity was found to have increased compared to vehicle-exposed control rings. Stimulation with Phe or Ach caused a small increase over basal NOS activity in these preparations. Prolonged exposure to Phe also caused an enhancement of Ach mediated vasorelaxation in rat aorta. 4 Immunoblot and reverse transcription polymerase chain reaction experiments showed that prolonged exposure of rat aorta to Phe resulted in an increased expression of eNOS, but not iNOS. This increase was antagonized by nonselective antagonist prazosin. Immunohistochemical staining experiments also showed that expression of eNOS increased in endothelial cells after Phe exposure of the aortas. 5 These results, all together, showed that prolonged exposure of rat aorta to alpha(l)AR agonist Phe enhanced the expression of eNOS and basal NOS activity, which probably causes a decreased vasocontractile response to Phe or to other agonists such as 5HT (5-hydroxytryptamine) in rat aorta. 6 This phenomenon can be considered more as a functional antagonism of vasocontractile response to agonists mediated by endothelium than a specific desensitization of alpha(1)AR-mediated signalling in vascular smooth muscle cells. PMID- 15753949 TI - Characteristics of attenuated endothelium-dependent relaxation seen in rabbit intrapulmonary vein following chronic nitroglycerine administration. AB - 1 This study was undertaken to determine whether long-term in vivo administration of nitroglycerine (NTG) downregulates the endothelium-dependent relaxation induced by acetylcholine (ACh) in the rabbit intrapulmonary vein and, if so, whether the type 1 angiotensin II receptor (AT(1)R) blocker valsartan normalizes this downregulated relaxation. 2 In strips treated with the cyclooxygenase inhibitor diclofenac, ACh induced a relaxation only when the endothelium was intact. A small part of this ACh-induced relaxation was inhibited by coapplication of two Ca(2+)-activated K(+)-channel blockers (charybdotoxin (CTX)+apamin) and the greater part of the response was inhibited by the nitric oxide-synthase inhibitor N(omega)-nitro-L-arginine (L-NNA). 3 The endothelium dependent relaxation induced by ACh, but not the endothelium-independent relaxation induced by the nitric oxide donor NOC-7, was significantly reduced in NTG-treated rabbits (versus those in NTG-nontreated control rabbits). The attenuated relaxation was normalized by coapplication of valsartan with the NTG. 4 In the vascular wall, both the amount of localized angiotensin II and the production of superoxide anion were increased by in vivo NTG treatment. These variables were normalized by coapplication of valsartan with the NTG. 5 It is suggested that long-term in vivo administration of NTG downregulates the ACh induced endothelium-dependent relaxation, mainly through an inhibition of endothelial nitric oxide production in the rabbit intrapulmonary vein. A possible role for AT(1)R is proposed in the mechanism underlying this effect. PMID- 15753951 TI - A selective novel low-molecular-weight inhibitor of IkappaB kinase-beta (IKK beta) prevents pulmonary inflammation and shows broad anti-inflammatory activity. AB - 1 Pulmonary inflammatory diseases such as asthma are characterized by chronic, cell-mediated inflammation of the bronchial mucosa. 2 Recruitment and activation of inflammatory cells is orchestrated by a variety of mediators such as cytokines, chemokines, or adhesion molecules, the expression of which is regulated via the transcription factor nuclear factor kappa B (NF-kappaB). 3 NF kappaB signaling is controlled by the inhibitor of kappa B kinase complex (IKK), a critical catalytic subunit of which is IKK-beta. 4 We identified COMPOUND A as a small-molecule, ATP-competitive inhibitor selectively targeting IKK-beta kinase activity with a K(i) value of 2 nM. 5 COMPOUND A inhibited stress-induced NF kappaB transactivation, chemokine-, cytokine-, and adhesion molecule expression, and T- and B-cell proliferation. 6 COMPOUND A is orally bioavailable and inhibited the release of LPS-induced TNF-alpha in rodents. 7 In mice COMPOUND A inhibited cockroach allergen-induced airway inflammation and hyperreactivity and efficiently abrogated leukocyte trafficking induced by carrageenan in mice or by ovalbumin in a rat model of airway inflammation. 8 COMPOUND A was well tolerated by rodents over 3 weeks without affecting weight gain. 9 Furthermore, in mice COMPOUND A suppressed edema formation in response to arachidonic acid, phorbol ester, or edema induced by delayed-type hypersensitivity. 10 These data suggest that IKK-beta inhibitors offer an effective therapeutic approach for inhibiting chronic pulmonary inflammation. PMID- 15753954 TI - Family and early life factors associated with changes in overweight status between ages 5 and 14 years: findings from the Mater University Study of Pregnancy and its outcomes. AB - OBJECTIVE: To describe different patterns of overweight status between ages 5 and 14 y and examine the role of modifiable family and early life characteristics in explaining different patterns of change between these two ages. DESIGN: A population-based prospective birth cohort. SUBJECTS: A total of 2934 children (52% males) who were participants in the Mater-University study of pregnancy, Brisbane, and who were examined at ages 5 and 14 y. MAIN OUTCOME MEASURES: Four patterns of change in overweight/obesity status between ages 5 and 14 y: (i) normal at both ages; (ii) normal at 5 y and overweight/obese at 14 y; (iii) overweight/obese at 5 y and normal at 14 y; (iv) overweight/obese at both ages. RESULTS: Of the 2934 participants, 2018 (68.8%) had a normal body mass index (BMI) at ages 5 and 14 y, 425 (14.5%) changed from a normal BMI at age 5 y to overweight or obese at age 14 y, 175 (6.0%) changed from being overweight or obese at age 5 y to normal weight at age 14 y and 316 (10.8%) were overweight or obese at both ages 5 and 14 y. Girls were more likely to make the transition from overweight or obese at age 5 y to normal at 14 y than their boy counterparts. Children whose parents were overweight or obese were more likely to change from having a normal BMI at age 5 y to being overweight at 14 y (fully adjusted RR: 6.17 (95% CI: 3.97, 9.59)) and were more likely to be overweight at both ages (7.44 (95% CI: 4.60, 12.02)). Birth weight and increase in weight over the first 6 months of life were both positively associated with being overweight at both ages. Other explanatory factors were not associated with the different overweight status transitions. CONCLUSIONS: Parental overweight status is an important determinant of whether a child is overweight at either stage or changes from being not overweight at 5 y to becoming so at 14 y. PMID- 15753952 TI - Mahanine, a carbazole alkaloid from Micromelum minutum, inhibits cell growth and induces apoptosis in U937 cells through a mitochondrial dependent pathway. AB - 1 Mahanine, a naturally occurring carbazole alkaloid in some Asian vegetables, has been shown to exhibit antimutagenicity, antimicrobial activity, cytotoxicity, and other biological properties. 2 In the present study, we investigated the effect of mahanine on the activation of the apoptotic pathway in human leukemia U937 cells. Various end points were used to screen for apoptosis: Morphological changes in cells, the relative numbers of viable and apoptotic cells; translocation of membrane bound phosphatidylserine and DNA analysis. 3 We found that mahanine-induced apoptosis in U937 cells involved activation of caspases, including caspase-3, release of cytochrome c into cytosol, loss of mitochondrial membrane permeability, and decreased levels of cellular ATP. 4 Pretreatment of cells with cyclosporine A, prior to/concomitant with exposure to mahanine, effectively prevented the deleterious effects of the alkaloid on cellular integrity and viability. 5 As mitochondrial permeability is known to be important in the regulation of cytochrome c release, our observations indicate that mitochondria are the principal target of mahanine. More specifically, we propose that mahanine causes the mitochondrial membranes to lose their permeability, resulting in caspase-3 activation and apoptosis. PMID- 15753953 TI - Prolactin stimulates integrin-mediated adhesion of circulating mononuclear cells to endothelial cells. AB - Attachment of leukocytes to endothelial cells is an essential step for the extravasation and recruitment of cells at sites of inflammation. The pituitary hormone prolactin (PRL) is involved in the inflammatory process. Here, we show that treatment with PRL of human peripheral blood mononuclear cells (PBMC) stimulates their adhesion to human umbilical vein endothelial cells (HUVEC) activated by interleukin-1beta. Stimulation of adhesion by PRL is mediated via integrins leukocyte functional antigen-1 (LFA-1) and very late antigen-4 (VLA-4), because immunoneutralization of both integrins prevents PRL action. Also, PRL promotes the adhesion of PBMC to immobilized intercellular adhesion molecule-1 and fibronectin, ligands for LFA-1 and VLA-4, respectively. Stimulation of integrin-mediated cell adhesion by PRL may involve the activation of chemokine receptors, because PRL upregulates the expression of the G-protein-coupled chemokine receptor CXCR3 in PBMC, and pertussis toxin, a specific G-protein inhibitor, blocks PRL stimulation of PBMC adhesion to HUVEC. In addition, PRL stimulates tyrosine phosphorylation pathways leading to leukocyte adhesion. PRL triggered the tyrosine phosphorylation of Janus kinase-2, of signal transducer and activator of transcription-3 and 5, and of the focal adhesion protein paxillin. Furthermore, genistein, a tyrosine kinase inhibitor, blocked PRL stimulated adhesion of PBMC and Jurkat T-cells to HUVEC. These results suggest that PRL promotes integrin-mediated leukocyte adhesion to endothelial cells via chemokine receptors and tyrosine phosphorylation signaling pathways. PMID- 15753955 TI - The importance of diagnostic criteria in the association between the metabolic syndrome and cardiovascular disease in obese subjects. AB - OBJECTIVE: To investigate whether the association between the metabolic syndrome (MS) and cardiovascular disease (CVD) in obese adults is influenced by the criteria used to diagnose the MS. DESIGN AND SUBJECTS: Cross-sectional study in 389 obese adults (male/female: 26%/74%; body mass index (BMI): 30.1-63.2 kg/m2; age: 18-79 y). MEASUREMENTS: To diagnose the MS by the WHO or the ATPIII criteria, body mass index, waist circumference, fasting and 2-h oral Glucose tolerance test plasma glucose, fasting plasma triglycerides and HDL cholesterol, systolic and diastolic blood pressure, 24-h albumin excretion, and fasting insulin were measured. The association between the MS diagnosed with either definition and self-referred CVD was investigated. RESULTS: The prevalence of the MS by the WHO was higher than by the ATPIII criteria (WHO 69.1%, ATPIII 49.4%; P<0.001). The MS diagnosed by the WHO criteria was significantly associated with self-referred CVD (odds ratio (OR) 5.80, 95% CI 1.35-24.95, P<0.05), whereas the ATPIIII MS was not (OR 1.34, 95% CI 0.59-3.03). An elevated blood pressure (OR 5.04, 95% CI 1.41-18.01, P<0.05) and microalbuminuria (OR 2.61, 95% CI 1.06-6.40, P<0.05) were independently associated with CVD. Consideration of the OGTT data as part of the ATPIII MS definition improved its associations with CVD (OR 4.39, 95% CI 1.29-14.94, P<0.05). CONCLUSION: The WHO criteria appear to identify a greater number of obese adults at risk for CVD. Nevertheless, the addition of an OGTT at least in nondiabetic patients with two ATPIII-defined metabolic risk factors may help to improve the association between the MS and CVD in obese adults. PMID- 15753956 TI - A genome scan in multigenerational families with dyslexia: Identification of a novel locus on chromosome 2q that contributes to phonological decoding efficiency. AB - Dyslexia is a common and complex developmental disorder manifested by unexpected difficulty in learning to read. Multiple different measures are used for diagnosis, and may reflect different biological pathways related to the disorder. Impaired phonological decoding (translation of written words without meaning cues into spoken words) is thought to be a core deficit. We present a genome scan of two continuous measures of phonological decoding ability: phonemic decoding efficiency (PDE) and word attack (WA). PDE measures both accuracy and speed of phonological decoding, whereas WA measures accuracy alone. Multipoint variance component linkage analyses (VC) and Markov chain Monte-Carlo (MCMC) multipoint joint linkage and segregation analyses were performed on 108 families. A strong signal was observed on chromosome 2 for PDE using both VC (LOD=2.65) and MCMC methods (intensity ratio (IR)=32.1). The IR is an estimate of the ratio of the posterior to prior probability of linkage in MCMC analysis. The chromosome 2 signal was not seen for WA. More detailed mapping with additional markers provided statistically significant evidence for linkage of PDE to chromosome 2, with VC-LOD=3.0 and IR=59.6 at D2S1399. Parametric analyses of PDE, using a model obtained by complex segregation analysis, provided a multipoint maximum LOD=2.89. The consistency of results from three analytic approaches provides strong evidence for a locus on chromosome 2 that influences speed but not accuracy of phonological decoding. PMID- 15753957 TI - Neurobiology of mood, anxiety, and emotions as revealed by studies of a unique antidepressant: tianeptine. AB - Recent studies have provided evidence that structural remodeling of certain brain regions is a feature of depressive illness, and the postulated underlying mechanisms contribute to the idea that there is more to antidepressant actions that can be explained exclusively by a monoaminergic hypothesis. This review summarizes recent neurobiological studies on the antidepressant, tianeptine (S 1574, [3-chloro-6-methyl-5,5-dioxo-6,11-dihydro-(c,f)-dibenzo-(1,2-thiazepine)-11 yl) amino]-7 heptanoic acid, sodium salt), a compound with structural similarities to the tricyclic antidepressant agents, the efficacy and good tolerance of which have been clearly established. These studies have revealed that the neurobiological properties of tianeptine involve the dynamic interplay between numerous neurotransmitter systems, as well as a critical role of structural and functional plasticity in the brain regions that permit the full expression of emotional learning. Although the story is far from complete, the schema underlying the effect of tianeptine on central plasticity is the most thoroughly studied of any antidepressants. Effects of tianeptine on neuronal excitability, neuroprotection, anxiety, and memory have also been found. Together with clinical data on the efficacy of tianeptine as an antidepressant, these actions offer insights into how compounds like tianeptine may be useful in the treatment of neurobiological features of depressive disorders. PMID- 15753958 TI - No evidence for association to the G72/G30 locus in an independent sample of schizophrenia families. PMID- 15753959 TI - Encephalomyelitis following mumps. AB - STUDY DESIGN: A case report with review of literature since 1961. SETTING: Departments of Neurology and Radiology, Faculty of Medicine, Zonguldak Karaelmas University, Zonguldak, Turkey. METHOD: A 22-year-old male patient diagnosed and treated as mumps encephalomyelitis is described. The clinical findings of the patient and the difficulties in differential diagnosis are discussed with the help of previously reported eight adult cases. RESULTS: Increased T2 signals in the spinal cord from C4 to C6 and T5 to T10 were seen by magnetic resonance imaging. This was the largest spinal involvement in the reported mumps cases. He was treated with supportive therapy oral steroids and early rehabilitation. On the sixth month, his neurological examination revealed bilateral 2/5 paresis in lower extremities. CONCLUSION: Encephalomyelitis following mumps is an uncommon but serious event in adults. PMID- 15753960 TI - Spinal injuries admitted to a specialist centre over a 5-year period: a study to evaluate delayed admission. AB - STUDY DESIGN: Retrospective study of 432 patients admitted to our institution with a spinal injury over a 5-year period. OBJECTIVES: To present epidemiological data relating to this spinal population, reporting specifically on delayed admission and length of hospitalisation. SETTING: Royal National Orthopaedic Hospital, Stanmore, UK. METHODS: A total of 432 traumatic spinal injuries admitted between March 1998 and March 2003 were analysed with respect to age, gender, mechanism of injury, level of bony injury, neurological level (complete, incomplete and intact), Injury Severity Score (ISS), date of injury, referral and admission independently and length of hospitalisation. The delays between injury and referral (>3 days) and between referral and admission (>7 days) were correlated to the length of hospitalisation. A detailed analysis of the cause of delay at both junctures was undertaken. RESULTS: There were 322 males (average age, 38.6 years) and 110 females (average age, 41.8 years) in our study. Classification of neurological severity disclosed 108 complete injuries, 115 incomplete and 209 intact. The average time between injury and referral was 5.5 days (range 0-94), and between referral and admission was 10.7 days (range 0 130). A total of 161 patients (37%) experienced a delay between injury and referral, of whom 59 (37%) were subsequently also delayed to admission. The principal reason for delay between injury and referral was the treatment of concurrent injuries. Even patients with complete injuries (15/43) experienced delayed referral. In all, 112 patients (26%) experienced a delay between referral and admission. Principal reasons included the provision of beds (Intensive care, acute and rehabilitation) and physiological stabilisation of other injuries particularly thoracic trauma. CONCLUSIONS: Provision of beds remains the most common preventable reason for delay between referral and admission and is associated with increased hospitalisation. Early liaison with a designated spinal injuries unit, particularly those with cord injury remains vitally important. PMID- 15753961 TI - In-patient medical resource utilization for high-level cervical spinal cord injury without bone fracture in Taiwan. AB - STUDY DESIGN: Retrospective review of secondary data obtained from the Bureau of National Health Insurance (BNHI) on medical resource utilization in in-patient cervical spinal cord injury patients in Taiwan. OBJECTIVES: Since the start of the National Health Insurance Program in Taiwan in 1995, costs have continued to increase each year. High-level cervical spinal cord injury, a catastrophic illness, consumes a large amount of medical resources. Appropriate control of in patient costs for these patients is mandatory. Analyses of the factors influencing the health-care costs of these patients are needed, so cost containment policies can be established by the BNHI to conserve health-care resources. SETTING: Health-care institutions throughout Taiwan. METHODS: We obtained secondary data on a randomized basis for diagnostic codes 952.00, 952.01, 952.02, or 952.03 of the International Classification of Diseases, Ninth Revision, Clinical Modification from the BNHI files of annual in-patient expenses during the period from 1998 to 2000. There were 184 hospital admission records studied. RESULTS: The lengths of stay and in-patient costs were significantly different among different hospital types. Length of stay also was statistically different according to patient, gender, and age. The lengths of stay and in patient costs were influenced by the hospital accreditation level and patient gender. Medical orders were influenced by patient age. CONCLUSIONS: Basic and selective diagnostics and therapeutics for high-level spinal cord injury without bone fracture should be established. Thus, patient needs for appropriate medical care will be met and overuse of medical resources will be prevented. Communication among doctors also should be strengthened. PMID- 15753962 TI - Primary osteosarcoma of the thoracic spine: report of an unusual elderly patient with autopsy findings. AB - STUDY DESIGN: A case report of primary osteosarcoma of the spine in an elderly patient. OBJECTIVE: The histopathological features of osteosarcoma vary widely, often leading to diagnostic difficulties particularly when there is little evidence of osteoid formation. The report describes the difficulty in the diagnosis of osteosarcoma of the thoracic vertebra. SETTING: Department of Orthopaedics and Rehabilitation Medicine, Fukui University Faculty of Medicine, Fukui, Japan. METHOD: A 78-year-old man presented with paraparesis and underwent urgent anterior excision of a primary spinal tumor emanating from the T10 vertebra followed by artificial vertebral replacement. The patient eventually died of disseminated disease of vertebral osteosarcoma. RESULTS: Samples from the T10 vertebral tumor showed neoplastic growth of atypical spindle-shaped cells, with foci of storiform-like proliferation. The tissue also demonstrated positive immunohistochemical staining for vimentin and alpha-smooth muscle actin and a tentative diagnosis of leiomyosarcoma was made. However, a metastatic nodule of the chest wall at autopsy showed focal osteoid formation, a finding not seen in the primary tumor. CONCLUSION: Early detection and accurate diagnosis is important for improving not only patient prognosis but also the quality of life. We should always consider this rare entity, particularly in elderly patients who present with back pain and vertebral collapse. PMID- 15753963 TI - Metabolic and cardiorespiratory responses of tetraplegic subjects during treadmill walking using neuromuscular electrical stimulation and partial body weight support. AB - STUDY DESIGN: Determination of differences in the cardiorespiratory responses of tetraplegic subjects with incomplete and complete lesions during treadmill gait and endurance exercise provided by neuromuscular electrical stimulation (NMES). Differences between rest and exercise phases were also examined. OBJECTIVES: To compare the cardiorespiratory responses in tetraplegic individuals during endurance exercise in the sitting position and treadmill gait with 30-50% body weight relief, both provided by NMES. SETTING: Rehabilitation Ambulatory at University Hospital, Brazil. METHODS: A total of 31 tetraplegic subjects were evaluated. Individuals were separated into two groups: gait group and endurance exercise group. In the gait group (n=17), the exercise protocol consisted of three different phases: 8 min of rest, 10 min of treadmill walking using NMES and 10 min of recovery. In the endurance exercise group (n=14), the cardiorespiratory test consisted of 8 min of rest, 15 min of quadriceps endurance exercise in the sitting position by NMES and 10 min of recovery. Oxygen uptake (VO(2)), carbon dioxide production (VCO(2)), respiratory exchange ratio (RER), pulmonary ventilation (V(E)) and heart rate (HR) were measured. RESULTS: All parameters increased considerably in the gait group from rest to the walking phase, although individuals with incomplete lesions presented a more pronounced increase than those with complete lesion. In the gait group, for incomplete tetraplegics, the mean VO(2) peak was 0.816+/-0.314 l/min, corresponding to 11.41+/-3.11 ml/kg/min; the mean value for VCO(2) was 0.660+/-0.24 l/min, mean HR was 124.54+/-28.72 bpm and mean V(E) was 28.38+/-6.28 l/min. In the endurance exercise group, for incomplete tetraplegics, the mean VO(2) peak was 0.246+/-0.07 l/min, corresponding to 3.84+/-0.92 ml/kg/min; the mean value for VCO(2) was 0.205+/ 0.06 l/min, mean HR was 71.45+/-15.51 bpm and mean V(E) was 11.83+/-2.72 l/min. In the endurance exercise group, smaller differences were observed in all variables from rest to the exercise phase, compared with the large increases observed in the gait group. CONCLUSIONS: These preliminary results have shown that gait training probably improves physical capacity in tetraplegic individuals more than the endurance exercise. PMID- 15753964 TI - Postmortem study of the spinal cord showing snake-eyes appearance due to damage by ossification of the posterior longitudinal ligament and kyphotic deformity. AB - CLINICAL DESIGN: A case report. OBJECTIVES: To elucidate the clinical role of snake-eyes appearance in this case, correlation between radiological, clinical and postmortem study was performed. SETTING: Aichi, Japan. CASE REPORT: A 73-year old man developed weakness and pain in the upper limbs due to kyphotic deformity secondary to laminectomy for cervical ossification of the posterior longitudinal ligament. Axial magnetic resonance imaging revealed snake-eyes appearance from C4 to C6. He died of acute myocardial infarction 3 months after anterior decompressive surgery. RESULTS: A postmortem examination of the cervical spinal cord showed small cystic six necrotic areas at the junction of the central gray matter and the ventrolateral posterior column, one in the right and one in the left, in association with neuronal loss in the anterior horn. CONCLUSIONS: Bilateral small intramedullary high-signal areas known as 'snake-eyes appearance' located around the central gray matter and the ventrolateral posterior column, are associated with neuronal loss in the compressed anterior horn that played an important role in worsening weakness of the upper limbs. PMID- 15753966 TI - Technological advances in adoptive immunotherapy. AB - Adoptive immunotherapy is an attractive and elegant strategy for treating a variety of life-threatening diseases. Several approaches have been developed to generate antigen-specific CD4+ and CD8+ T cells for adoptive T-cell therapy in cancer and infectious diseases. Currently, many approaches are based on either the use of autologous peptide pulsed dendritic cells as antigen-presenting cells or nonspecific expansion of T cells. Unfortunately, current approaches lack the ability to serve as reproducible and economically viable methods. Several groups are developing new artificial approaches to overcome problems associated with dendritic cells and the nonspecific expansion of T-cell clones in order to make adoptive immunotherapy more feasible and effective. Thus, by increasing the availability of adoptive immunotherapy, we will be able to better determine the efficacy of the approaches in the treatment of a variety of diseases. In this review, we focus on technological advances that will facilitate adoptive immunotherapy. Specifically, we summarize current strategies which are either based on artificial antigen-presenting cells or on T-cell receptor gene transfer. PMID- 15753965 TI - Second-generation antipsychotic medications in the treatment of mood disorders: focus on aripiprazole. AB - Second-generation antipsychotic medications offer a broader range of therapeutic efficacies than first-generation agents. Consequently, our field has witnessed a rapid expansion of the use of second-generation antipsychotic drugs for several conditions beyond psychosis. The use of second-generation antipsychotic medications has been most pronounced in mood disorders, especially in bipolar disorders. Information about the agents clozapine, risperidone, olanzapine, quetiapine, ziprasidone and aripiprazole in terms of their efficacy and tolerability in bipolar disorder is now available. Aripiprazole, a new agent whose proposed mechanism(s) of action differs from that of other agents, has been shown in placebo-controlled comparative trials in bipolar patients to be an effective and well tolerated treatment option for this patient group. The role of second-generation antipsychotic medications in the therapeutic armamentarium for bipolar disorders will be determined by clinical experience, by additional phase IV studies and by trials that compare second-generation antipsychotics with each other and also with mood-stabilizing medications. There is also a pressing need for additional information on the long-term efficacy and safety of each second generation antipsychotic agent during maintenance therapy for bipolar disorders. PMID- 15753967 TI - Bevacizumab: antiangiogenic cancer therapy. AB - Angiogenesis, the process of new blood vessel formation, is an essential step in tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is a key mediator in this process, and elevated levels of this cytokine are observed in solid tumors and are correlated with worse clinical outcomes. Research has therefore focused on developing agents that target angiogenic factors such as VEGF in order to inhibit tumor growth. One such agent is bevacizumab, a humanized monoclonal antibody generated by engineering the VEGF-binding residues of a murine neutralizing antibody into the framework of a normal human immunoglobulin G. Bevacizumab recognizes VEGF receptors 1 and 2 and thus can neutralize the biologically active forms of VEGF that interact with these receptors. In addition, bevacizumab has shown antiangiogenic and antitumor activity in several cancer types, recently gaining approval from the FDA for use in combination with fluorouracil-based chemotherapy as a first-line treatment for metastatic cancer of the colon or rectum. PMID- 15753968 TI - The causes of skin cancer: a comprehensive review. AB - Skin cancer is the most common type of cancer in fair-skinned populations around the world. The incidence and mortality rates of skin cancers are dramatically increasing and thus pose a threat to public health. Understanding the etiology and pathogenesis of skin cancer remains a goal for healthcare systems. A clearer understanding of causative factors is an essential step in the prevention of skin cancer. This article comprehensively reviews the causative agents which play a role in the development of skin cancer. Ultraviolet radiation (UV) from sun exposure is the most important cause of skin cancer. Sunburns and excessive exposures cause cumulative damage which induces immunosuppression and skin cancers. Ozone depletion, the level of UV light, elevation, latitude, altitude and weather conditions influence the emission of UV radiation reaching the earth's surface. Organ transplant recipients and AIDS patients have an increased incidence of skin cancers. Some treatment modalities, including radiation therapy, phototherapy and psoralen and long-wave ultraviolet radiation (PUVA) can also predispose to skin cancers. Viral infections such as the human papilloma virus can cause squamous cell carcinomas. Individuals with familial genetic syndromes are susceptible to specific types of skin cancers. Ionizing radiation, environmental pollutants, chemical carcinogens and work-related exposures have been associated with skin cancers. Exposure to artificial UV radiation (tanning beds and lamps), aging, skin color, diet and smoking are attributable risks. Skin cancers have been found in dermatoses and various types of keratoses, chronically injured or nonhealing wounds, and scars. This article provides a comprehensive and thorough overview of skin cancer, with an emphasis on understanding its epidemiology, incidence, etiology and related risk factors. PMID- 15753969 TI - Antihypertensive drug interactions. AB - There are many drug interactions with antihypertensive agents and some of these are highly significant. Patients with hypertension frequently take multiple medications and may be at increased risk for drug interactions. Nearly every elderly patient with multiple medical problems will have the potential for one drug interaction in their regimen. These drug interactions can lead to morbidity or even mortality if appropriate steps are not taken to minimize this risk. Drug interactions may occur due to pharmacokinetic (i.e., absorption, distribution, metabolism, elimination) or pharmacodynamic interactions. Physicians and pharmacists must remain vigilant in their monitoring of potential drug interactions and make appropriate dosage or therapy adjustments. PMID- 15753970 TI - The impact of vascular risk factors on erectile function. AB - The main cause of erectile dysfunction is organic in nature, with vascular etiologies being the most common risk factors. The incidence of erectile dysfunction increases with the number of vascular comorbidities such as hypertension, diabetes, hyperlipidemia, smoking and atherosclerosis. The earliest signal of endothelial damage in men with vascular risk factors is the manifestation of erectile dysfunction. The penis is a barometer of the body's endothelial function, so it is reasonable then to correlate vascular pathologies as direct causes of erectile dysfunction. Moreover, erectile dysfunction may be the first clinical presentation of any of these comorbidities, with the vascular endothelium playing a pivotal role in regulating vascular homeostasis of the corpora cavernosa. This article addresses the impact of vascular risk factors on erectile function based on current evidence. PMID- 15753972 TI - Oxidative damage and Alzheimer's disease: are antioxidant therapies useful? AB - Oxidative stress is a key factor involved in the development and progression of Alzheimer's disease, and it is well documented that free radical oxidative damage, particularly of neuronal lipids, proteins, nucleic acids and sugars, is extensive in brains of Alzheimer's disease patients. However, oxidative stress may elicit compensatory responses and downstream adaptations such as amyloid-beta deposition and neurofibrillary tangle formation, which may function as "shields" to ensure that neuronal cells do not succumb to oxidative injuries. Although during the past several years our understanding of the mechanisms leading to neuronal damage and death in the course of Alzheimer's disease has improved significantly, we have not found an effective therapeutic to fight this devastating disorder. However, the results obtained in clinical trials with antioxidants are promising and propel us in the search of new and more effective antioxidant therapies. PMID- 15753971 TI - Biphasic expression of lipin suggests dual roles in adipocyte development. AB - The identification of gene mutations that cause lipodystrophies, conditions characterized by a lack of normal adipose tissue, has revealed new proteins that play a role in adipocyte biology. Lipin is one such protein identified in a lipodystrophic mouse strain and found to be critical for normal adipocyte differentiation. Interestingly, lipin displays a biphasic expression pattern in adipocytes, with peaks of expression at two points during adipogenesis--a transient induction in preadipocytes prior to expression of peroxisome proliferator-activated receptor gamma, and a second wave of expression in mature adipocytes. Thus, lipin appears to have critical roles in both adipocyte differentiation and in the function of mature adipocytes. PMID- 15753973 TI - The year's new drugs. AB - Twenty-three new biological drugs and diagnostic agents reached their first markets in 2004. The oncolytic drugs therapeutic group was the most active in terms of new launches, with seven market introductions, and the United States was the most active single market for new products, with a total of 10 new launches in 2004. PMID- 15753974 TI - Molecule of the month. LBM-415. PMID- 15753975 TI - Fighting to get into the restaurant. AB - Although perhaps not the highest profile AIDS conference in the world, the Seventh International Congress on Drug Therapy in HIV Infection, held November 14 18, 2004, in Glasgow, Scotland, had a unique profile. It consisted entirely of plenary sessions devoted to drug treatment. This saved delegates from having to rush from one venue to another and allowed people to concentrate on a single topic. The fact that the congress has been held in this format since 1992 is a strong confirmation that the organizers have found a winning formula. All but one of these congresses have been held in Glasgow, and it has been very interesting to see how attitudes have changed over the years. This seventh congress attracted more than 2,500 delegates from 69 countries. Highlights of the meeting are discussed in this report. PMID- 15753976 TI - CNS drug discovery: challenges and solutions. AB - The worldwide market for therapies for central nervous system (CNS) disorders was valued at around US dollars 50 billion in 2001, and is set to grow sharply in the years ahead. This is because of a marked increase in the number of people aged over 65 (the "baby boomer" effect), which will lead to increased demand for more safe and effective medicines for CNS disorders. This one-day Society for Medicines Research symposium, held September 23, 2004, in London, United Kingdom, was organized by Dr. Alan M. Palmer (Pharmidex, London, U.K.) and Prof. F. Anne Stephenson (School of Pharmacy, University of London, U.K.). More than 100 delegates heard a scholarly and comprehensive review of the challenges currently facing CNS research and development, which was accompanied by consideration of a variety of innovative solution strategies. The meeting considered: 1) how to identify and validate targets for potential CNS drugs; 2) how to assess brain penetration (both in vitro and in vivo); 3) how to develop in silico methodologies to predict blood-brain barrier penetration; 4) how to assess therapeutic efficacy (both in vitro and in vivo); 5) how to establish reliable biomarkers to guide decision making; and 6) how to effectively apply magnetic resonance imaging to CNS drug discovery. PMID- 15753977 TI - Recent advances in heart research. AB - Of the many symposia at the 28th World Congress of the International Society of Heart Research, held August 7-10, 2004, in Queensland, Australia, this report covers the following: understanding the basic mechanisms of cardiac arrhythmias; unstable plaque; innate immunity in cardiovascular health and disease; matrix metalloproteinases; defining the targets for treatment of heart failure; NAD(P)H oxidase-derived reactive oxygen species--signaling or stress; expanding roles for alpha(1)-adrenoceptors in the cardiovascular system; estrogen derivatives in cardiovascular disease--old questions, new answers; and p38 MAPK: a kind or callous kinase. Lectures on the use of gene therapy for cardioprotection and antiarrhythmic actions of calmodulin kinase inhibition, and selected poster presentations are also discussed. PMID- 15753978 TI - Manganese superoxide dismutase overexpression changes plating efficiency bidirectionally according to change in redox for SaOS2 human osteosarcoma cell line. AB - Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that converts cytotoxic superoxide radicals into hydrogen peroxide. MnSOD activity is lower in tumor cells, and MnSOD overexpression reportedly ameliorates malignant phenotypes. We established stable MnSOD overexpressing cell lines from a human osteosarcoma cell line, SaOS2, and then investigated the effects of MnSOD overexpression on plating efficiency (PE) and the involvement of reactive oxygen species, including nitric oxide (NO) in those effects. The PE of SaOS2FM(L), a moderate MnSOD overexpression cell line, increased, while that of SaOS2FM(H), a high MnSOD overexpression cell line, decreased. Although we assessed PE using a colony-formation assay, time-lapse microscopic observation revealed that cells attached to the flasks had undergone neither apoptosis nor necrosis. Moreover, MnSOD overexpression did not affect cell doubling time. Therefore, MnSOD overexpression might correlate directly with cellular adhesion's effect on PE changes. When L-buthionine-[S,R]-sulfoximine (BSO) was administered to increase the intracellular concentration of hydrogen peroxide, the PEs of both cell lines decreased, and when hydrogen peroxide was eliminated by the administration of sodium pyruvate, only the PE of SaOS2FM(H) increased. The combination of BSO and NO (NOR4 or isosorbide 5-mononitrate) administration synergistically decreased PE in both cell lines. These findings suggest that changes in cellular adhesion properties correlate with the balance between increased hydrogen peroxide levels and decreased superoxide radical levels. This is the first report to indicate that PE and cellular adhesion properties change bidirectionally according to the levels of MnSOD overexpression: first increasing then decreasing as MnSOD activity increases. Our results indicate that PE changes might be decided by the balance between two cytotoxic compounds (decreased superoxide radical levels and increased hydrogen peroxide levels), and that NO loading and increased hydrogen peroxide synergistically reduce PE and cellular adhesion. PMID- 15753979 TI - Activation of p53/p21Waf1/Cip1 pathway by 5-aza-2'-deoxycytidine inhibits cell proliferation, induces pro-apoptotic genes and mitogen-activated protein kinases in human prostate cancer cells. AB - The tumor suppressor gene p53 plays an essential role in cell proliferation and apoptosis. Due to its relevance to cancer therapy, most studies have focused on the cellular consequences of p53 activation in relation to cytotoxic drugs. 5-aza 2'-deoxycytidine (5-aza-CdR) is widely used as an anti-cancer drug for the treatment of leukemia and solid tumors. However, the mechanism by which 5-aza-CdR exerts its anti-neoplastic activity remains unclear. Here, we address the role of p53 in regulating cellular responses to 5-aza-CdR treatment in human prostate cancer cells. We found that 5-aza-CdR induces p53 and p21Waf1/Cip1 expression associated with inhibition of cell proliferation in LNCaP cells (p53 wild-type), but not in DU145 cells (p53 mutant). By using pifithrin-alpha, a chemical inhibitor of p53, we confirmed that the increase in p21Waf1/Cip1 expression and inhibition of cell proliferation in LNCaP cells by 5-aza-CdR is p53-dependent. Also, the activation of p53 and p21Waf1/Cip1 pathway by 5-aza-CdR modified multiple gene expressions including apoptotic target genes and MAP kinases in LNCaP cells. 5-aza-CdR-induced apoptosis in LNCaP cells is assessed by DNA fragmentation analysis. Furthermore, knockdown of p53 by pU6-p53 siRNA vector suggests the involvement of MAP kinases in the process of 5-aza-CdR-mediated activation of p53 pathway to inhibit cell proliferation and induce apoptosis. Finally, the comet or SCGE assay and methylation-sensitive restriction analysis demonstrated that 5-aza-CdR induced p53 and p21Waf1/Cip1 expression as a consequence of DNA damage and independent of DNA demethylation. Our findings suggest that 5-aza-CdR induces anti-neoplastic activity primarily through the activation of p53 pathway in response to DNA damage and subsequently leads to inhibition of cell proliferation as well as induction of apoptosis. Therefore, our data indicate that p53 status in tumor cells may be critical for the clinical efficacy and toxicity of 5-aza-CdR. PMID- 15753980 TI - Different expression levels of lumican in human carcinoid tumor and neuroendocrine cell carcinoma. AB - Lumican is a member of the small leucine-rich proteoglycan (SLRP) family and participates in the maintenance of tissue structures and tumor growth. Neuroendocrine cell tumors (NETs) including carcinoid tumors and NE cell carcinomas (NECs) possess numerous neuroendocrine (NE) granules, and differences between these tumors are in terms of their biological and metastatic aggressiveness during tumor progression. The purpose of this study was to examine the expression of lumican in NETs, and to determine whether the presence of lumican may be associated with the growth of NETs by comparing its expression between carcinoid tumors and NECs. Immunohistochemically, the positivity rates of lumican expression in the cytoplasm of the tumor cells were 87.5% in carcinoid tumors and 37.5% in NECs. Those of lumican expression in the stroma adjacent to the tumors were 90.1% in carcinoid tumors and 79.2% in NECs. In situ hybridization analysis revealed the lumican mRNA expression in the cytoplasm of carcinoid tumor and NEC cells. Ultrastructurally, the lumican protein was observed in the rough endoplasmic reticulum and NE granules of NETs and interspaces of collagen fibers. Furthermore, RT-PCR analysis revealed the presence of lumican mRNA in NEC cell lines. These results indicate that the higher expression level of cytoplasmic lumican in carcinoid tumors than in NECs may play a role in the slow growth of these tumors. PMID- 15753981 TI - Inhibition of tumor growth and angiogenesis by water extract of Gac fruit (Momordica cochinchinensis Spreng). AB - The antitumor activity of the crude water extract from Gac fruit (Momordica cochinchinensis) was investigated in vivo and in vitro. A water extract prepared from 0.75 and 0.25 mg dry weight of Gac fruit per gram body weight was given daily to Balb/c mice (n=15/group). The water extract inhibited the growth of the colon 26-20 adenocarcinoma cell line, transplanted in Balb/c mice, reducing wet tumor weight by 23.6%. Histological and immunohistochemical results indicated that Gac water extract reduced the density of blood vessels around the carcinoma. The water extract also produced a marked suppression of cell proliferation in colon 26-20 and HepG2 cells. Cell cycle analysis demonstrated a significant accumulation of cells in the S phase by water extract. Immunoblotting showed that cyclin A, Cdk2, p27waf1/Kip1 were down-regulated, whereas the protein level of p21waf1/Cip1 was not decreased. Treatment of colon 26-20 cells with Gac extract induced necrosis rather than apoptosis. The antitumor component was confirmed as a protein with molecular weight of 35 kDa, retained in the water-soluble high molecular weight fraction. Thus, the bioactive antitumor compound in Gac extract is a protein, which is distinct from lycopene, another compound in Gac fruit with potential antitumor activity. PMID- 15753982 TI - Aberrant expression of the gastric mucin MUC6 in human pulmonary adenocarcinoma xenografts. AB - Certain pulmonary adenocarcinomas show gastrointestinal differentiation with the expression of various mucins. The CDX homeobox gene, an intestine-specific transcription factor, is related to gastric carcinogenesis with MUC2 and MUC6 expression. The intestinal mucin MUC2 is expressed in the normal lung, while the gastric mucin MUC6 is not. Previously, we have reported that the expressions of MUC2 and MUC6 were related to a poor prognosis in small adenocarcinomas of the lung. We estimated the expressions of the mucin (MUC2 and MUC6) and CDX (CDX1 and CDX2) to examine how CDX relates to the gastrointestinal mucin production in the pulmonary adenocarcinoma. Thirty-nine human non-small cell lung cancer (NSCLC) xenografts were examined (13 adenocarcinoma, 18 squamous cell carcinoma and 8 large cell carcinoma). Significant expression of the MUC6 gene was observed in 7 out of 39 (17.9%) NSCLC xenografts. The expressions of the MUC6 genes were noted in 6 out of 13 (46.2%) adenocarcinoma xenografts, but only in 1 of 18 (0.06%) squamous cell carcinoma xenografts. The adenocarcinoma xenografts significantly showed higher expression of the MUC6 gene than squamous cell carcinoma xenografts (t-test, p=0.0343). Four adenocarcinoma-xenografts co-expressed both the MUC2 and MUC6 genes, and the residual 2 adenocarcinoma-xenografts expressed only the MUC6 gene. One MUC6 overexpressing squamous cell carcinoma focally contained an adenocarcinoma component. The expression patterns of the gastrointestinal mucins were analogous to gastric cancer. The cellular morphology of these carcinoma xenografts was of the gastric cancer type. The proteins of the MUC2 and MUC6 were immunohistochemically confirmed in the xenografts. The expression of the MUC6 gene was significantly correlated with the expressions of the CDX1 and CDX2 genes in the xenografts (Fisher's test, p<0.0001 and p=0.0005, respectively), while there was no significant association between the expression of the MUC2 and CDX genes. These results suggest that the expression of CDX molecules in the pulmonary carcinogenesis pathway relates to gastric cancerous features of aberrant MUC6 expression. PMID- 15753983 TI - GM1 expression in H-ras-transformed NIH3T3 results in the suppression of cell proliferation inducing the partial transfer of activated H-ras from non-raft to raft fraction. AB - c-H-ras is located in lipid/rafts and undergoes cholesterol dependent regulation. To analyze the regulatory effects of ganglioside GM1 on the proliferation of fibroblasts transformed with mutated ras gene, GM1 synthase cDNA was transfected into NIH3T3/H-ras cells containing mutation. In the transient expression system with EGFP-fused GM1 synthase, the ratio of BrdU-positive cells among EGFP positive cells was compared between GM1(+) transfectant cells and control cells, indicating that the transient GM1 expression suppresses cell proliferation. Then, established transfectant cells C21 and C34 expressed definite levels of GM1, and analyzed for the cell growth with the control cells D2 and D4 expressing no GM1. GM1(+) cells showed reduced proliferation compared with controls. Phosphorylation levels of ERK1/2 after FCS treatment were examined, showing that those on the GM1(+) transfectant cells increased slowly, while those in the controls rapidly reached the plateau. Fractionation of Triton X-100 extracts with sucrose density gradient ultra-centrifugation revealed that activated H-ras proteins from controls as well as NIH3T3/H-ras were completely localized in non-GEM/raft fraction. On the other hand, some portions of activated H-ras were transferred to GEM/raft fraction, i.e., 32% in C21, and 8% in C34. Since the Ras effector Raf-1 was localized in non-GEM/raft, the growth suppression might be caused, at least partly, by the movement of activated H-ras to GEM/raft fraction. PMID- 15753984 TI - Enhanced expression of p46 Shc in the nucleus and p52 Shc in the cytoplasm of human gastric cancer. AB - Shc protein is known to be related to cell proliferation and carcinogenesis. However, the involvement of Shc in gastric cancer remains unknown. In the present study, we examined the expression and localization of Shc in human gastric cancer and the adjacent normal mucosa by Western blotting and immunohistochemical analyses. Furthermore, the expression of Shc in a gastric cancer cell line, MKN 28, was also studied. p66 Shc was not detected at all in gastric cancer or the adjacent normal mucosa. In contrast, immunohistochemical and Western blot analyses revealed that p52 Shc was detected in the cytoplasmic fractions obtained from gastric normal mucosa and cancer, while p46 Shc expression was observed in the nuclear fractions from gastric normal mucosa and cancer. Furthermore, not only p52 Shc expression in the cytoplasm but also p46 Shc expression in the nucleus was much higher in gastric cancer than in the adjacent normal mucosa. Subsequent in vitro experiments with MKN 28 gastric cancer cells also revealed that p52 Shc was expressed solely in the cytoplasm and p46 Shc was solely in the nucleus. These results suggest that the enhancement of p46 Shc and p52 Shc expression may be related to the occurrence of gastric cancer. Furthermore, unlike p52 Shc, p46 Shc was shown to be expressed solely in the nucleus, suggesting that p46 Shc expressed in the nucleus may play an important role in gastric carcinogenesis. PMID- 15753985 TI - Beta catenin is associated with breast cancer progression in vitro. AB - Cancer is induced by a series of genetic alterations that lead to changes in the normal mechanisms controlling cell proliferation, differentiation, cell death, or genomic instability. The MCF-10F, a spontaneously immortalized human breast epithelial cell line, treated with benzo(a)pyrene and then transfected with the c Ha-ras oncogene was used in these studies. The aim was to define the phenotypic alterations associated with the carcinogenic process. Carcinogen-treated and c-Ha ras-transfected cells showed a progression of changes in the morphology as seen by transmission electron microscopy, anchorage-independent growth, invasiveness and capability of tumor formation in the SCID mice, as well as altered oncoprotein expression. Furthermore, these cells showed an increased expression of MDM2, Int-2 (FGF-3) and beta catenin in comparison to control MCF-10F as determined by fluorescence staining coupled with confocal microscopy. The MDM2, Int-2 (FGF-3) expressions were increased in cell lines transfected with the c-Ha ras with or without carcinogen treatment as well as the tumor cell line derived from a tumor formed in the SCID mice in comparison to control cell line MCF-10F. However, beta catenin was only increased in the most aggressive tumorigenic cell lines in comparison with MCF-10F cell line and non-transfected ones. It can be concluded that malignant progression is a stepwise process and tumor growth occurs after a series of molecular events that parallel morphological changes indicative of cell transformation. PMID- 15753987 TI - Clinicopathological relevance of HER2/neu and a related gene-protein cubic regression correlation in colorectal adenocarcinomas in Taiwan. AB - While HER2/neu receptor tyrosine kinase is involved in various malignancies, studies on colorectal adenocarcinoma (CRC) remain controversial. To try to clarify the role played by HER2/neu in CRC, sixty-seven CRC patients in Taiwan were analyzed. For this analysis, we used normalized dual-color fluorescence in situ hybridization (FISH) and Photoshop-aided immunohistochemistry (IHC) between cancers and their autologous non-neoplastic epithelia. The results revealed that HER2/neu status was unrelated to age, sex, location and positive-nodal percentage. Intramucosal carcinomas had earlier HER2/neu protein upregulation than regional stromal invasion within Dukes' A, and had a gene level that had not risen yet. Both gene gains and protein increases were significant in later stages in regards to volumetric progression and nodal-metastatic Dukes' stage. Overall, there were 1.53-fold (gene) and 1.81-fold (protein) increases from non-neoplastic enterocytes to CRCs. The upregulating directions of gene (88%) and protein (88%) presented symmetric agreement. Most CRCs exhibited low to intermediate levels of HER2/neu overexpression with double-minute gene amplicons and cytosolic HER2/neu proteins. Normalized FISH and IHC showed high cubic-regression correlation, especially in Dukes' C. According to the correlation curve, the points with IHC index >2.41 and FISH ratio >1.22 defined the area where gene-amplification dependent HER2/neu overexpression was present. Eleven (16%) patients had values above the cut-off point (IHC = 2.41 and FISH = 1.22), including 7 (10%) cases in cytosolic and 4 (6%) cases in membranous HER2/neu overexpressions. The results suggest that HER2/neu plays a crucial role in CRC tumorigenicity with gene amplification-independent transcriptional activations early in the carcinogenesis, and gene-amplification-dependent overexpression later in the advanced stages. This indicates that HER2/neu can be a good biological marker for selecting patients that may improve under therapies that employ adequate HER2/neu targeting strategies. PMID- 15753986 TI - Altered expression of members of the IGF-axis in clear cell renal cell carcinoma. AB - Renal cell carcinoma (RCC) is a heterogeneous disease and its biology is poorly understood. The commonest subtype identified is clear cell RCC. The insulin-like growth factor axis is intimately involved with many cellular roles including that of renal development. Dysregulation of this axis has frequently been demonstrated in cancer. In this study, we examine the expression of several IGF-axis components, including receptors, ligands, and binding proteins in clear cell renal cell carcinoma. A series of clear cell RCCs with matched normal kidney from the same individuals were obtained. Total RNA was extracted and expression levels of genes examined using RNase protection analysis. We confirm the dysregulation of the IGF-axis within clear cell renal cell carcinoma including the upregulation of IGFBP-3, which is further validated by immunohistochemical staining on a tissue array containing 50 RCC: positive staining in 29/30 clear cell; 1/10 papillary and 0/10 chromophobe. In addition, we demonstrate that the expression of the A isoform of the insulin receptor is significantly upregulated, while that of IGFBP-5 are significantly downregulated in this tumour subtype. PMID- 15753988 TI - Aggressive forms of non-Hodgkin's lymphoma in two patients bearing coinfection of Epstein-Barr and hepatitis C viruses. AB - Although epidemiologic and experimental data suggest an etiopathogenetic role for both hepatitis C virus (HCV) and Epstein-Barr virus (EBV) infection in development of B-cell non-Hodgkin's lymphoma (NHL), potential interactions between EBV and HCV during progression of B-cell NHL have not yet been fully investigated. In the present study, tumor biopsy specimens from patients with both B-cell NHL and chronic HCV infection (HCV(+)) were analyzed for the presence of EBV-encoded RNA (EBER) by in situ hybridization (ISH). VH and VL gene segments were amplified from tumor biopsy specimen DNA by PCR. EBV infection (EBV(+)) was detected in tumors from 2 of 31 (6%) HCV(+) B-cell NHL patients. Clinical histories of these two EBV(+)/HCV(+) B-cell NHL patients indicated a particularly aggressive course of disease. Chemotherapy failed to induce long lasting remission for either of these EBV(+)/HCV(+) B-cell NHL patients. Amplification of CDR3 of the Ig heavy chain gene from DNA isolated from each EBV(+)/HCV(+) B-cell NHL indicated the presence of monoclonal B-cell expansion. Rearrangement of Ig genes in neoplastic B-cell clones from both EBV(+)/HCV(+) patients was similar to that previously reported for EBV(-)/HCV(+) B-cell NHL patients. Additionally, neoplastic B-cell clones from these two EBV(+)/HCV(+) B-cell NHL patients did not exhibit intraclonal variation. Previous studies have demonstrated that intraclonal variation is common among neoplastic B-cell clones from EBV(-)/HCV(+) patients. EBV infection may have prevented evolution of variant neoplastic B-cell clones by suppressing antibody affinity maturation. Together, these data suggest that EBV infection may cooperate with HCV infection during progression of B-cell NHL in immunocompetent individuals. Such an interaction may accelerate the course of disease in B-cell NHL patients. PMID- 15753989 TI - Overexpression of Endothelin-A-receptor in breast cancer: regulation by estradiol and cobalt-chloride induced hypoxia. AB - Previous studies have demonstrated the potential significance of Endothelin (ET) 1 and its receptors, ETAR and ETBR, in the development and progression of breast cancer. The objective of this study was to assess the expression levels and potential regulation of the "ET axis" in human non-neoplastic and neoplastic breast tissue as well as in various human breast cancer cell lines. Expression of ET-1, ETAR and ETBR was evaluated in 31 neoplastic and 7 non-neoplastic breast tissue samples and in six human breast cancer cell lines using conventional and quantitative real-time RT-PCR, Western blotting and immunohistochemistry. The effects of 17beta-estradiol (E2) and cobalt-chloride (CoCl2) treatment on ET-1, ETAR and ETBR expression were studied in vitro. ETAR mRNA expression levels were found to be statistically significantly higher in breast cancer specimens than in non-neoplastic breast tissue (p<0.001). For ET-1 and ETBR mRNA expression, no significant difference was observed between the two groups. All cell lines exhibited expression of ET-1 and ETAR mRNA, whereas none showed significant ETBR mRNA expression. We observed a strong and reproducible induction of ETAR mRNA and protein expression by E2 and CoCl2 in MDA-MB-468 and BT-474 cells and in MDA-MB 453 and SK-BR-3 cells with a maximum increase after 8 and 16 h of treatment, respectively, while MCF-7 and HBL-100 cells showed a constitutive expression pattern. The present data suggest a novel mechanism in the regulation of ETAR expression in breast cancer. Based on these findings, a combination of ETAR antagonists with adjuvant endocrine treatment seems to be a reasonable therapeutic strategy. PMID- 15753990 TI - Establishment of human fibroma cell lines from a MEN1 patient by introduction of either hTERT or SV40 early region. AB - Establishment of tumor cell lines as model systems for studying tumor biology or as a part of immunotherapeutic anti-cancer strategies is of high importance, whereby the highest possible preservation of the original tumor cell phenotype is a prerequisite for these aims. Since overexpression of the catalytic subunit of human telomerase (hTERT) is known to minimally alter the cellular phenotype, we focused on the establishment of cell lines derived from human fibroma from a MEN1 patient by ectopic expression of hTERT. Additionally, a cell line was generated by introduction of the early region of SV40 (SV40 ER). Both approaches resulted in continuous cell lines, and neither T1-LOHG (hTERT) nor SV1-LOHG (SV40 ER) showed a transformed phenotype. While SV40 ER-transfected cells underwent dramatic changes in morphology and growth characteristics, hTERT-expressing cells indeed retained a phenotype highly similar to the parental cells. Nevertheless, hTERT overexpression resulted in increased growth rates after about 70 population doublings (PD) and alterations of mRNA levels of genes associated with tumor pathogenesis. Thus, our data suggest that ectopic hTERT expression leads to immortalization of LOHG-F, sustaining many characteristics of the non-transfected counterparts, but continuous growth in vitro is associated with changes of the cellular phenotype. PMID- 15753991 TI - Cancer disease predictive diagnosis: BAT/CD3-positive lymphocytes in cancer patients. AB - BAT is an immune-activating monoclonal antibody produced against Daudi cell membranes and selected for stimulating lymphocyte proliferation. The anti-tumor activity of BAT is related to its immunostimulatory properties. Both T and NK cells mediate the anti-tumor activity of BAT. CD4-positive T cells respond to BAT activation by proliferation and INF-gamma production. The aim of the study was to assess the probability that the BAT monoclonal antibody binding capacity to T cells is a marker for different cancers. Human peripheral blood T cells from colon, breast and prostate cancer patients, as well as healthy volunteer donors, were tested for the percentage of binding to BAT mAb (BAT/CD3 cells) by FACS analysis. All patients were tested before undergoing surgery or treatment, and their diagnosis was confirmed by histology. The results showed that the percentage of BAT monoclonal antibody binding to CD3-positive T cells in the peripheral blood was different in cancer patients with diverse tumor types. We found that lymphocytes from the blood of healthy donors contained 25% BAT/CD3 cells. In colon and breast cancer patients, a significant decrease to 13 and 11% of BAT/CD3 cells was found. In contrast, these cells increased ><50% in patients with prostate cancer. These findings may have a potential diagnostic significance and also assist in the evaluation of strategies for the therapeutic use of BAT for different cancer patients. PMID- 15753992 TI - Comparison of the prognosis indication of VEGFR-1 and VEGFR-2 and Tie2 receptor expression in breast carcinoma. AB - The degree of angiogenesis in breast cancer has previously been shown to be an indicator of prognosis, and tumor microvasculature is a candidate target for new antiangiogenic therapies. The aim of this study was to investigate the prognostic value of vascular endothelial growth factor (VEGF) receptors, VEGFR-1 (Flt-1) and VEGFR-2 (KDR/Flk-1), and Tie2/tek receptor tyrosine kinase in breast carcinoma. VEGF receptors and Tie2 expression was investigated using immunohistochemical assays with monoclonal antibodies on frozen sections in a series of 918 and 909 patients respectively. VEGFR-1 and VEGFR-2 and Tie2 were correlated with long term (median, 11.3 years) patients' outcome. Univariate (Kaplan-Meier) analysis showed that VEGFR-1 positive tumor surface (cutoff = 5%) was significantly correlated with high metastasis risk (p=0.03) and relapse (p<0.01) in all patients, and in those with node negative tumors (p<0.001 and p<0.01 respectively), but not with overall survival. In contrast Tie2 positive tumor surface (cutoff = 7%) was significantly correlated with poor overall survival (p=0.025) and also with high metastasis risk particularly among node negative patients (p<0.01). Moreover, Tie2 immunoexpression was significantly predictive of relapse (p=0.003) in the node negative subgroup (p=0.02). In multivariate analysis (Cox model), VEGFR-1 and Tie2 immunoexpressions were identified as independent prognostic indicators. In contrast, univariate analysis showed that VEGFR-2 positive tumor surface (cutoff = 10%) was not correlated with survival or with metastasis and relapse risk. Our results suggest that VEGFR-1 and Tie2 immunohistochemical expression permits the identification of patients with poor outcome, and particularly node negative ones with a high risk for metastasis and relapse. VEGFR-1 and Tie2 immunodetection may also be considered as potential tools for selecting patients who could benefit in the future from specific antiangiogenic therapy interfering with VEGFR-1 and Tie2 activation pathways. PMID- 15753993 TI - Methylthioadenosine phosphorylase gene is silenced by promoter hypermethylation in human lymphoma cell line DHL-9: another mechanism of enzyme deficiency. AB - Methylthioadenosine phosphorylase (MTAP) involved in the metabolism of purine and polyamine has been known to be deficient in a variety of tumors. Although this enzyme deficiency was reportedly caused by partial or total deletion of the MTAP gene, human MTAP-deficient lymphoma cell line DHL-9 has the intact MTAP gene. In order to determine the mechanism of MTAP deficiency in DHL-9, we carried out methylation-specific PCR analysis of sodium bisulfite-treated genomic DNA followed by DNA sequence analysis. Following incubation with various concentrations of 5-Aza-2'-deoxycytidine, DHL-9 cells were subjected to RT-PCR and an immunoblot analysis for MTAP expression. MTAP promoter in DHL-9 cells was methylated at cytosine of all CpG dinucleotides analyzed. Moreover, 5-Aza-2' deoxycytidine treatment induced DHL-9 cells to express MTAP mRNA and protein. Taken together, MTAP deficiency in DHL-9 was caused by transcriptional silencing due to promoter methylation. Promoter methylation of the MTAP gene was also found in DNA samples from adult T-cell leukemia patients. These results indicated that promoter hypermethylation is another mechanism of MTAP deficiency in human malignancy. Thus, immunological diagnostics will be needed for an accurate evaluation of MTAP expression at the protein level. PMID- 15753994 TI - Proteome-based identification of molecular markers predicting chemosensitivity to each category of anticancer agents in human gliomas. AB - To identify the protein markers that are clinically useful for predicting efficacy of anticancer agents, we investigated the correlation between the proteome profiling patterns and the in vitro chemosensitivity in human gliomas. The proteome of 93 surgical samples were analyzed with two-dimensional gel electrophoresis (2DE) and mass spectrometry. The in vitro chemosensitivities to 10 different kinds of anticancer agents (cyclophosphamide, nimustine, cisplatin, cytosine arabinoside, mitomycin C, peplomycin, adriamycin, etoposide, vincristine, paclitaxel) were measured by flow cytometric detection of apoptosis. We identified a set of 41 proteins that significantly affected the in vitro chemosensitivity to each category of anticancer agents. Many of the proteins that correlated with chemoresistance were categorized into the signal transduction proteins including the G-proteins. The present study showed that the proteome analysis using 2DE could provide a list of proteins that may be the potential predictive markers for chemosensitivity in human gliomas. They can also be direct and rational targets for anti-glioma therapy and be used for sensitization to the conventional chemotherapeutic regimens. PMID- 15753995 TI - Inhibition of choline kinase renders a highly selective cytotoxic effect in tumour cells through a mitochondrial independent mechanism. AB - Tumour cells are frequently altered in their phospholipid metabolism. Choline kinase (ChoK, E.C. 2.7.1.32) activity, the first enzyme involved in the synthesis of phosphatidylcholine, is increased in a large number of human tumours and tumour-derived cell lines. We have previously reported that MN58b, a specific inhibitor of ChoK, has anti-tumoral activity. Here we show the high specificity of MN58b as a cytotoxic drug towards tumour cells, and explore further the basis of its mechanism of action in order to provide a rational understanding for its antitumoral activity. A dramatic difference in the response to the treatment of primary, normal and non-tumorigenic human cells when compared to tumour-derived cell lines was observed. In normal cells, blockage of de novo PCho synthesis by MN58b results in a reversible cell cycle arrest at G0/G1 phase. In contrast, ChoK inhibition in tumour cells promotes the induction of apoptosis. This effect depends on the cell cycle phase, being G1 the critical phase. Regarding the mechanism of apoptosis engagement, a loss of mitochondrial potential was observed 10-20 min after cytochrome c release, but caspase 3 activation preceded the loss of mitochondrial potential, indicating that activation of caspase 3 is independent of cytochrome c release. Our results are consistent with a non intrinsic process as the mechanism underlying the induction of apoptosis by ChoK inhibition in tumoral cells. PMID- 15753996 TI - Expression of human telomerase reverse transcriptase splice variants is well correlated with low telomerase activity in osteosarcoma cell lines. AB - The human telomerase reverse transcriptase (hTERT) gene is a catalytic subunit of telomerase that plays an important role in maintaining telomere length. Recently, post-transcriptional (alternative splicing) and epigenetic (promoter methylation) mechanisms affecting expression of the hTERT gene have been reported as negative regulators of telomerase activity (TA). To elucidate the significance of the telomere maintenance mechanism (TMM) in osteosarcoma (OS), we examined hTERT mRNA/protein expression and methylation status in 16 OS cell lines and compared them with TA. Five (31%) of 16 OS cell lines expressed both full-length and splice variants of hTERT mRNA/protein, and 3 (19%) exhibited only full-length hTERT mRNA/protein. No hTERT mRNA/protein expression was observed in 8 cell lines (50%). TA in OS cell lines exhibiting full-length hTERT mRNA/protein without any splice variants was higher than in cell lines expressing splice variants. The promoters of the 16 OS cell lines were relatively hypermethylated, but no inverse relationship between frequency of methyl-CpG sites and hTERT expression was observed. Treatment with a demethylating agent induced hTERT mRNA/protein in only one cell line. These results suggest that the epigenetic mechanism might contribute to the regulation of the hTERT gene in a small subset of OSs, and that alternative splicing might be involved in controlling the TA of OS cell lines, thereby contributing to their TMM. PMID- 15753997 TI - Growth inhibition of A549 human lung carcinoma cells by beta-lapachone through induction of apoptosis and inhibition of telomerase activity. AB - The objective of the present study was to investigate the effect of beta lapachone, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae), on the cell growth and apoptosis in human lung carcinoma cell line A549. Exposure of A549 cells to beta-lapachone resulted in growth inhibition and induction of apoptosis in a time- and dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometry analysis. This increase in apoptosis was associated with a decrease in Bcl-2 and expression, an increase of Bax, and an activation of caspase-3 and caspase-9. beta-lapachone treatment markedly inhibited the activity of telomerase in a dose-dependent fashion. Additionally, the levels of human telomerase RNA (hTR) and c-myc expression were progressively down-regulated by beta-lapachone treatment. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of beta-lapachone. PMID- 15753998 TI - Rationale for sequential tamoxifen and anticancer drugs in adjuvant setting for patients with node- and receptor-positive breast cancer. AB - Since the survival benefit of tamoxifen (TAM) combined with anticancer drugs in treating node- and receptor-positive breast cancer is small, appropriate treatment schedules and the rationale for the combination remains unclear. We examined the effect of estradiol (E2) on sensitivity to anticancer drugs to clarify the survival benefit of tamoxifen combined with anticancer drugs. We used the MTT assay to assess the effect of E2 on sensitivity to anticancer drugs in the E2 receptor-positive and -negative breast cancer cell lines, MCF-7 and MDA-MB 231, respectively. We assessed the expression of apoptosis-related proteins by Western blotting, and evaluated apoptosis using the TUNEL method. Serum levels of E2 were measured using an enzyme-labeled radioimmunoassay in patients with premenopausal breast cancer before and during treatment with tamoxifen. Estrogen administration decreased sensitivity in MCF-7 cells to the anticancer drugs, adriamycin (ADM), mitomycin C (MMC), and paclitaxel (TXL), evaluated as increases in the IC50 values for ADM (4.1-fold), MMC (1.9-fold) and TXL (13.0-fold), compared with those of each drug alone. Estradiol in MDA-MB-231 cells similarly increased the IC50 values for ADM (9.5-fold), MMC (15.6-fold), and TXL (2.4 fold). The decreased sensitivity to these anticancer drugs was associated with the attenuation of apoptosis. Estrogen dose-dependently increased the expression of Bcl-2 protein in MCF-7, but not in MDA-MB-231 cells, and suppressed the expression of Bax and cytochrome c induced by anticancer drugs in association with decreased apoptosis compared with the effect of each drug alone. Phosphorylation of the Bcl-2 protein induced by TXL was decreased in the presence of E2 in MCF-7 cells. Serum levels of E2 were increased in 5 patients without amenorrhea and in 1 patient with amenorrhea after treatment with TAM alone in adjuvant therapy, compared with levels before treatment. Estradiol decreased sensitivity to ADM, MMC, and TXL in MCF-7 and MDA-MB-231 breast cancer cells, and this was associated in part with an increase in the amount of Bcl-2 protein, and decreases in levels of Bax and cytochrome c leading to apoptosis. These results suggest that therapy with TAM and anticancer drugs should be sequentially scheduled with anticancer drugs followed by TAM in an adjuvant setting to treat patients with breast cancer for a potentially improved survival benefit. PMID- 15753999 TI - Effects of WNT/beta-catenin pathway activation on signaling through T-cell factor and androgen receptor in prostate cancer cell lines. AB - Dysregulation of the WNT/beta-catenin pathway is thought to contribute to prostate cancer progression. Mutations of beta-catenin occurring in 5-7% of advanced prostate cancers may act by stimulating TCF-dependent and/or androgen receptor (AR)-dependent transcription. Using a reporter gene approach we found overexpressed mutated beta-catenin to enhance AR-regulated probasin-promoter activity in the AR-positive prostate cancer cell line 22Rv1, particularly at low androgen levels. In 22Rv1 cells mutated beta-catenin was able to stimulate TCF dependent transcription but was unable to do so in LNCaP cells where it activates the AR. Since beta-catenin mutations are rare in vivo, we studied further possible routes of WNT-pathway modulation. Higher concentrations of LiCl, a GSK3beta-inhibitor, were required to activate TCF-dependent rather than AR dependent reporter constructs. In 22Rv1 overexpression of E-cadherin repressed androgen-dependent transcription, but did not inhibit transcription of TCF dependent reporter genes as in bladder cancer cell lines. Interestingly, Wnt-3a stimulated proliferation selectively in the AR-positive prostate cancer cell lines 22Rv1 and LNCaP, even though TCF-dependent reporter gene transcription was not induced in LNCaP cells. In summary, the data from our study support the idea that activation of WNT/beta-catenin signaling in AR-positive prostate cancer cells may predominantly act through AR-dependent mechanisms rather than classical TCF-dependent mechanisms. PMID- 15754000 TI - Retinoid receptor mRNA expression profiles in human bladder cancer specimens. AB - Retinoids, which include vitamin A (retinol) and its derivatives, have previously been investigated as potential chemopreventive and chemotherapeutic agents in bladder cancer. We examined mRNA expression of the retinoid receptors RARalpha, RARbeta2, RARgamma and RXRalpha, as well as two putative RARbeta2 target genes, DAB2 and Midkine, in normal and malignant bladder tissue specimens from human patients. We evaluated 24 normal and malignant bladder specimens for retinoid receptor, DAB2 and Midkine mRNA expression using RT-PCR. We also examined the effects of retinoic acid and retinol on the expression of these genes in five human bladder cancer cell lines. Expression of RARalpha, RARbeta2, RARgamma and RXRalpha mRNA was detected in all of the non-neoplastic patient bladder specimens. RARbeta2 mRNA expression was undetectable in 7/13 tumors, RARalpha in 3/13, RARgamma in 1/13 and RXRalpha in 2/13. DAB2 mRNA was expressed in all non neoplastic and all tumor specimens, while Midkine mRNA was detected in 8/11 non neoplastic specimens versus 11/13 tumors. Two of the five bladder cancer cell lines expressed RARbeta2 independent of retinoid exposure; in three cell lines RARbeta2 expression was induced by retinoids. RARalpha, RARgamma and RXRalpha mRNA expression was detected in 5/5 cell lines, independent of retinoid exposure. We found a reduction in retinoid receptor mRNA expression, particularly for RARbeta2, in human bladder cancer specimens. We also demonstrated induction of RARbeta2 mRNA expression in some of the retinoid-treated bladder cancer cell lines. We suggest that restoration of RARbeta2 expression may be a reasonable biomarker for developing bladder cancer preventive and/or therapeutic drugs. PMID- 15754001 TI - Antisense-thioredoxin inhibits angiogenesis via pVHL-mediated hypoxia-inducible factor-1alpha degradation. AB - Thioredoxin (Trx), a small redox protein, is overexpressed in a number of tumors, however, its roles in tumor cells were not defined well. To investigate the effect of Trx, we transfected Trx or antisense Trx vectors into HT1080 cells. Trx overexpressed HT1080 cells induced migration of endothelial cells through Flt-1 vascular endothelial growth factor (VEGF) receptor type-1. However, the migration was reduced by overexpression of antisense-Trx. To understand the relationship between Trx and hypoxia-induced angiogenesis, we observed the expression level of VEGF and hypoxia-inducible factor-1alpha (HIF-1alpha) after transfection of Trx. Overexpression of Trx also caused a significant increase of VEGF in protein and RNA levels under normoxic and hypoxic conditions. Moreover, transfection of Trx caused a dramatic increase in HIF-1alpha protein level under hypoxic condition. However, transfection of antisense-Trx markedly decreased HIF-1alpha and VEGF expression compared with controls. In addition, HIF-1alpha was not translocated from cytoplasm to nucleus in antisense-Trx overexpressing HT1080 cells. Further, we could detect the association of HIF-1alpha and pVHL in antisense-Trx transfectant HT1080 cells. Taken together, we suggest that Trx plays an important role in angiogenesis and, therefore, antisense-Trx might be applicable to the inhibition of tumor angiogenesis through the induction of pVHL-mediated HIF 1alpha degradation. PMID- 15754002 TI - Aberrant expressions of annexin A10 short isoform, osteopontin and alpha fetoprotein at chromosome 4q cooperatively contribute to progression and poor prognosis of hepatocellular carcinoma. AB - Chromosome 4q exhibits high frequency of allelic loss in hepatocellular carcinoma (HCC). This study aimed to elucidate the interaction of the frequent aberrant mRNA expression of alpha-fetoprotein (AFP), osteopontin (OPN) and a novel short isoform of annexin A10 (ANXA10S) at 4q in the tumor progression among 294 patients who received surgical resection of unifocal primary HCC. AFP overexpression, OPN overexpression and ANXA10S down-regulation correlated with high-grade and high-stage tumors, early tumor recurrence (all P<0.0001), and lower 10-year survival (all P=0.000001). The AFP overexpression correlated with OPN overexpression (P=0.0026) and ANXA10S down-regulation (P=0.00001), while OPN overexpression correlated with ANXA10S down-regulation (P=0.00001). Pair-wise combinations revealed interactive effects between these genetic variants for tumor grade, tumor stage and early recurrence (all P<0.0001). HCCs with more genetic aberrations had more frequent high tumor grade, portal vein invasion (stage IIIB-IV) and early recurrence (all P<0.0001). The 10-year survival rate for HCCs with all three genetic alterations was the lowest (7%), followed by those with two (22%) or one event (29%), and the highest for those without these changes (43%), P=0.000001. The prognostic stratification using these molecular factors was similar to that of histopathological staging. These three genetic alterations also helped to identify different subgroups of patients of stage II HCC but with different prognosis (P=0.015). In conclusion, the aberrant expressions of AFP, OPN and ANXA10S cooperatively contribute to tumor progression and poor prognosis, and are useful for molecular staging of HCC and the subclassification of stage II HCC without vascular invasion. PMID- 15754003 TI - Regulatory roles of altered N- and O-glycosylation of CD45 in galectin-1-induced cell death in human diffuse large B cell lymphoma. AB - To clarify the functions of CD45 N- and O-glycans in diffuse large B cell lymphoma (DLBCL), we analyzed the antiproliferative effects of bovine galectin-1 (beta-galactoside-binding lectin-1), which reacts with CD45 N-glycans and O glycans, on human lymphoma cells. Bovine galectin-1 induced cell death of the DLBCL cell line HBL-2 in vitro. Swainsonine (SW) is a potent inhibitor of alpha mannosidase II which catalyzes the synthesis of complex type N-linked oligosaccharides, and benzyl-GalNAc (BZGalNAc) is a potent inhibitor of O glycosylation. Treatment with SW or BZGalNAc prevented cell death of HBL-2 cells by galectin-1. Western blot analysis revealed SW treatment reduced the molecular weight by about 5 kDa of one isoform at 190 kDa among three isoforms of CD45 which have N-linked oligosaccharide ligands for galectin-1. BZGalNAc treatment reduced the molecular weight of another isoform by about 15 kDa. These data suggest that the amount of CD45 N-glycans or O-glycans was reduced by SW and BZGalNAc treatment, respectively, and that reduction of CD45 N-glycans or O glycans may prevent the interaction between CD45 and galectin-1. Alteration in CD45 N-glycans or O-glycans may regulate cell death of lymphoma cells through the interaction between CD45 N-glycans or O-glycans and galectin-1 in DLBCL. PMID- 15754004 TI - Uncoupling of oxidative phosphorylation and Smac/DIABLO release are not sufficient to account for induction of apoptosis by sulindac sulfide in human colorectal cancer cells. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) have shown chemopreventive properties in colorectal cancer, involving both cyclooxygenase (COX)-dependent and -independent mechanisms. Apart from their selectivity for COX isoenzymes, NSAIDs differ in their acidic character which supports ability to uncouple oxidative phosphorylation. To assess the possible contribution of uncoupling to their antineoplastic properties, we compared the effect of sulindac sulfide (SS), an acidic NSAID and NS-398, a non-acidic tricyclic, on mitochondrial function and apoptosis in colorectal cancer cell lines (HT29, Caco-2, HCT15 and HCT116). Although cell lines displayed a different COX status, SS and NS-398 caused growth arrest in a dose-related manner. High dose (10(-4)M) of SS but not of NS-398, increased the percentage of subG1 cell population while reducing mitochondrial transmembrane potential (DeltaPsim). Cyclosporin A (CsA, 1 microM) prevented collapse of DeltaPsim induced by 10(-4)M SS but not by 7.5 microM FCCP used as a protonophoric control. SS and FCCP increased the cytosolic release of Smac/DIABLO which was differently affected by CsA pretreatment depending on the uncoupler. Finally, 7.5 microM FCCP failed to induce apoptosis whereas CsA prevented apoptosis induced by SS from 16% in HCT15 to 41% in HCT116. The present study shows that despite the ability of sulindac sulfide to behave as a protonophoric uncoupler, CsA-sensitive opening of mitochondrial permeability transition pore contributes little to its pro-apoptotic effect in colorectal cancer cells. PMID- 15754005 TI - Altered expression of the hormone- and xenobiotic-metabolizing sulfotransferase enzymes 1A2 and 1C1 in malignant breast tissue. AB - Cytosolic sulfotransferases (SULTs) catalyze the biotransformation of steroid hormones as well as drugs and environmental toxins. Mostly, sulfonation leads to an inactivation of parent compounds, although formation of more toxic and cancerogenic metabolites also occurs. To assess possible alterations in the SULT enzyme expression pattern between malignant and non-malignant tissue, we studied the presence of 9 SULT enzymes of family 1 and 2 by semi-quantitative RT-PCR. Forty-two specimens from ductal and lobular breast carcinomas, lymph node metastasis, mastopathy and normal breast tissue were derived from 29 patients. Substantial expression of SULT 1A1, 1A2, 1A3, 1B1, 1C1, 1E1, 2A1, 2B1a and 2B1b mRNAs was observed in malignant and non-malignant tissue, although the pattern of the individual SULTs varied between the patients, and SULT1C1 mRNA was present in a greater number of malignant than non-malignant tissues (p<0.05). A major finding was that unspliced SULT1A2 mRNA, containing the complete intron between exons 7 and 8, was found in 4 of 16 non-malignant specimens, but was undetectable in the 26 malignant samples investigated. Taken together, the presence of various SULT enzymes in normal, premalignant and malignant breast tissue suggests an important role of SULT-mediated biotransformation in the breast. While the increased expression of SULT1C1 in malignant tissue seems to reflect tumor dedifferentiation, our finding of unspliced SULT1A2 mRNA in non-malignant tissue offers additional aspects regarding the search for breast cancer risk factors. PMID- 15754006 TI - Systemic delivery of RafsiRNA using cationic cardiolipin liposomes silences Raf-1 expression and inhibits tumor growth in xenograft model of human prostate cancer. AB - Raf-1, a protein serine-threonine kinase, plays a critical role in mitogen activated protein kinase kinase (MKK/MEK)- mitogen-activated protein kinase (extracellular signal-regulated kinase) (MAPK/ERK) pathways. We show here that systemically delivered novel cationic cardiolipin liposomes (NeoPhectin-AT) containing a small interfering RNA (siRNA) against Raf-1 silence the expression of Raf-1 in tumor tissues and inhibit tumor growth in xenograft model of human prostate cancer. The knockdown of Raf-1 expression by siRNA is also associated with down-regulation of cyclin D1 expression in vivo. PMID- 15754007 TI - Localised non-Hodgkin lymphoma of the testis: the Sheffield Lymphoma Group experience. AB - We evaluated clinical features, management and survival of patients with localised (stage 1E or 2E) testicular non-Hodgkin lymphoma (NHL) presenting to the Sheffield Lymphoma Group between 1972 and 2002. Thirty consecutive eligible patients were identified from the lymphoma database and clinical records of all were reviewed. Survival data were statistically analysed. The median age of patients was 74 years (range 38-87). Sixty-three percent were >70 years of age. The median duration of follow-up was 15 months (range 2.1-211). All patients had orchidectomy. Eleven (37%) had orchidectomy alone. Twelve (40%) had orchidectomy and chemotherapy, 4 (13%) had orchidectomy and radiotherapy. Three (10%) had orchidectomy, chemotherapy and radio-therapy. Five (17%) had immediate disease progression following orchidectomy or whilst on treatment. A complete response was seen in the remaining 25 (84%) irrespective of treatment modality. Seven (23%) of patients remain alive, with a median follow-up of 73 months (range 9.9 211). Six (20%) died of causes not related to their lymphoma. Twelve (40%) relapsed following a complete response and median time to relapse was 9 months. Only 1 of these relapses was in the central nervous system; 1 in bone, 1 in skin, 2 in the contralateral testis. The majority of relapses (58%) occurred in the lymph nodes. Primary testicular lymphoma is an uncommon and poor prognosis disease usually affecting older men. The data and experience available to guide the treatment strategies for testicular lymphoma are limited. However, the evidence points to a combined modality approach being the most likely effective treatment, comprising surgery, anthracycline-containing combination chemotherapy with or without prophylactic contralateral testis and regional lymph node irradiation. PMID- 15754008 TI - Genistein suppresses the invasive potential of human breast cancer cells through transcriptional regulation of metalloproteinases and their tissue inhibitors. AB - Progression of breast cancer implicates the degradation of extracellular matrix (ECM) by metallo-proteinases (MMPs), a process with important consequences on the growth and invasiveness of cancer cells in adjacent and distant sites. The isoflavone, genistein--a natural inhibitor of protein tyrosine kinase pathway- inhibits the growth of a wide range of cancer cells in vitro. The aim of this study was to investigate: i) the expression of mRNAs encoded for MMPs and their endogenous inhibitors (TIMPs) associated with pathogenesis and metastatic potential of breast cancer cells; and ii) the effect of genistein on the transcription of MMPs and TIMPs and the invasive potential of breast cancer cells. Gene expression at transcriptional level was examined in cell cultures of two epithelial breast cancer cell lines, the high invasive (ER-negative) MDA-MB 231 and the low invasive (ER-positive) MCF-7, as well as the normal mammary cells (MCF-12A) following RNA isolation and reversed transcriptase polymerase chain reaction (RT-PCR). The inhibitory effect of genistein on functional invasiveness was examined by a cell invasion assay. Cell cycle distribution showed that genistein arrested breast cancer MDA-MB-231, MCF-7 and BT-20 cells in the G2/M phase. Both normal and breast cancer cell lines express the genes of MMP-2, -9, MT1-, MT2-, MT3-MMP and TIMP-1, -2 and -3. MCF-7 express notably less MMPs than MDA-MB-231 cell line. The addition of genistein resulted in down-regulation of the transcription of all MMP genes in MDA-MB-231 and most of MMPs in MCF-7 cells. The inhibitory effect of genistein on MMPs was functionally confirmed, since it significantly reduced the invasion properties of cancer cells in vitro. The obtained results indicate that genistein may be of great value in prevention of breast cancer cell metastasis, since it represents both a transcriptional modulator of genes involved in this pathogenetic process and a suppressor of breast cancer cell invasiveness. PMID- 15754009 TI - The der(12)t(12;16) breakpoint in an acute leukaemia case targets a Sec7 domain containing protein. AB - A balanced translocation t(12;16)(p13;p13) in a patient with AML-M1 and bone marrow eosinophilia was previously analysed by FISH. The 16p13 breakpoint was shown to occur in the MYH11 gene, the fusion partner of CBFbeta in leukaemia patients with an inv(16) or a t(16;16), whereas the 12p13 breakpoint was shown to be present in cosmid c4H9. We present the molecular analysis of c4H9, resulting in the identification of a novel gene, SLAG. At the N-terminus, SLAG contains a Sec7 domain also found in proteins of the cytohesin family. In contrast to the cytohesins, no pleckstrin homology domain is present in SLAG. database searches identified several homologues, suggesting that SLAG defines a new subfamily of Sec7 proteins, characterised by an N-terminal Sec7 domain and a new C-terminal pleckstrin homology-like domain. The FISH data led to the hypothesis that rearrangement of SLAG might be involved in the pathogenesis of AML through the generation of a new fusion gene with MYH11. However, the putative SLAG-MYH11 fusions showed only weak transforming properties in NIH3T3 cells in a focus formation assay. PMID- 15754010 TI - Methylation of the CA9 promoter can modulate expression of the tumor-associated carbonic anhydrase IX in dense carcinoma cell lines. AB - CA IX is a tumor-associated transmembrane isoform of the carbonic anhydrase with a high enzyme activity and a functional involvement in the pH regulation and cell adhesion. Expression of CA9 gene in tumor cells is principally regulated by the high cell density and the hypoxia-related VHL-HIF pathway. In renal cell carcinomas with VHL inactivation, CA9 transcription is further controlled by site specific promoter methylation. Here we explored a possible role of methylation in the non-RCC cell lines represented mainly by HeLa cervical carcinoma cells. Using metabisulfite sequencing and treatment with the methylation inhibitor 5-aza-2' deoxycytidine we showed that the methylation of a single CpG site at -74 position with respect to the transcription start can down-modulate the expression of CA9 in cells cultivated at high density, but not in cells grown in sparse culture nor in cells exposed to hypoxia. Methylation appears to act in tumor cells expressing intermediate levels of CA IX protein, but not in cell lines expressing high CA IX levels. Our results indicate that promoter methylation is not crucial for the control of CA9 gene expression in the non-RCC cell lines but could represent an accessory mechanism restricting its expression in highly dense carcinoma cell cultures. PMID- 15754011 TI - Comparative genomics on Wnt8a and Wnt8b genes. AB - WNT family ligands transduce signals through FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, LRP5, LRP6, ROR1, ROR2 and RYK. WNT1, WNT2, WNT2B, WNT3, WNT3A, WNT8A, WNT8B, WNT10A and WNT10B are canonical WNTs to activate WNT - beta catenin pathway. Human WNT8A mRNA is expressed in NT2 cells with neuronal differentiation potential, while human WNT8B mRNA in diffuse type gastric cancer. Here, we identified and characterized the rat Wnt8a and Wnt8b genes by using bioinformatics. The rat Wnt8a gene, consisting of six exons, was located within AC134361.2 genome sequence. The rat Wnt8b gene, consisting of six exons, was located within AC105487.6 and AC103018.7 genome sequences. The rat Wnt8a (355 aa) and Wnt8b (350 aa) with 60.0% total-amino-acid identity were secreted-type proteins with 22 conserved Cys residues and two Asn-linked glycosylation sites. Wnt8b orthologs were more conserved than Wnt8a orthologs. GATA-binding site was located within conserved region of rat Wnt8b and human WNT8B promoters. GATA6 ESTs were expressed in diffuse type gastric cancer, and FGFR2 gene is reported preferentially amplified in diffuse type gastric cancer. KGF-FGFR2-PI3K-GATA6 WNT8B signaling cascade is predicted to play important roles in diffuse type gastric cancer. This is the first report on the rat Wnt8a and Wnt8b genes as well as on the conserved GATA-binding site within rat Wnt8b and human WNT8B promoters. PMID- 15754012 TI - Molecular evolution of Wnt2b orthologs. AB - WNT family proteins transduce signals through Frizzled (FZD) receptors with extracellular WNT-binding domain and cytoplasmic Dishevelled-binding domain. WNT2B-ST7L and WNT2-ST7 loci are paralogous regions within the human genome. WNT2B isoform 2 and WNT2 are canonical WNTs to activate the beta-catenin - TCF pathway in human cancer, especially in gastric cancer. Here, the rat Wnt2b gene was identified and characterized by using bioinformatics. The rat Wnt2b gene, consisting of five exons, was located within AC106372.5 genome sequence. Wnt2b core promoter region (nucleotide position 234449-233652 of AC106372.5 rat genome sequence) was identified as an evolutionarily conserved region. GC content of rat Wnt2b promoter was 71.8%. Nucleotide identity between human and rat Wnt2b promoters was 71.2%. E box, CP2-binding site, CCAAT box and ZNFN1A1 (IKAROS) binding site were located within the conserved region of rat Wnt2b, mouse Wnt2b, and human WNT2B promoters. Rat Wnt2b (391 aa), human WNT2B isoform 2 and other vertebrate Wnt2b orthologs were type II transmembrane proteins with cleavable extracellular WNT domain, while vertebrate Wnt2 orthologs were secreted proteins. Twenty-four Cys residues were conserved among vertebrate Wnt2b and Wnt2 orthologs. Glycosylation sites at Asn 283 of rat Wnt2b and RGD motif were identified within mammalian Wnt2b orthologs, but not within vertebrate Wnt2 orthologs and zebrafish wnt2b. Zebrafish wnt2b acquired N-terminal cytoplasmic region to evolve into type II transmembrane protein, and then mammalian Wnt2b orthologs acquired integrin-targeting RGD motif within the extracellular WNT domain. This is the first report on the rat Wnt2b gene as well as on molecular evolution of the Wnt2b orthologs. PMID- 15754013 TI - Relevance of nuclear localization and functions of RNA helicase A. AB - RNA helicase A (RHA) regulates gene expression at transcriptional and post transcriptional levels. It is also known that RHA shuttles between the nucleus and the cytoplasm. RHA possesses nuclear transport domain which functions as a nuclear localization signal (NLS) and nuclear export signal (NES), and predominantly localizes in the nucleus. Here, we identified regions that influence cellular localization of RHA. We showed that double-stranded RNA binding domain I and II (dsRBD I and II) positively localized to the cytoplasm. Mutational analysis revealed that RNA-binding ability was important to localization of dsRBD to the cytoplasm. RHA mutants lacking association with Pol II complexes were partially spilled from the nucleus. Furthermore, a minimal transactivation domain (MTAD), which is minimal region to interact with Pol II complexes, mainly localized to the nucleus in comparison with GFP. Our results indicated that the intracellular localization of RHA might be regulated by not only NLS and NES but also functional domains. PMID- 15754014 TI - Evolution of interleukin-18 binding proteins and interleukin-1 receptor, type II proteins. AB - Interleukin-18 (IL-18) is one of the pivotal cytokines controlling the defense mechanism called inflammation. As a first step to develop proteins for controlling the IL-18 level, we initiated a study of IL-18-binding proteins (IL 18BPs). Twenty-four IL-18BP family members, 11 from vertebrates and 13 from chordopoxviruses, were picked from the NCBI database. Eight of these vertebrate IL-18BPs and two of the chordopoxvirus IL18-BPs were identified here and characterized as new members of the IL-18BP family. Their IL-18 binding domains were aligned and the distribution of highly conserved critical amino acid residues was analyzed and used to construct a phylogenetic tree. From this tree it was inferred that at least two independent events created two different ancestral viral IL-18BP genes by retroposition of IL-18BP genes from the vertebrate lineage. These two events are estimated to have occurred after an ancient mammalian IL-18BP gene diverged from birds, and before the mammalian IL 18BP gene diverged into human, ungulate and rodent IL-18BP genes. Moreover, our results suggest that IL-18BP and interleukin-1 receptor, type II (IL-1R2) had a common ancestral gene and diverged from the ancestral gene into IL-18BP and IL 1R2 genes in the fish period. PMID- 15754015 TI - Adrenomedullin, ANP and BNP are colocalized in a subset of endocrine cells in the rat heart. AB - We investigated by immunocytochemistry (ICC) the distribution in the rat heart of adrenomedullin (AM), a potent and long-lasting hypotensive peptide which is expressed in the cardiovascular system, where it is known to play a major regulatory role. Hearts were collected from adult male Sprague-Dawley rats, and were perfused for 20 min, according to the Langendorff technique, with endothelin 1 (ET-1) or the mast cell-degranulator compound 48/80. Hearts were frozen, and ICC was performed using standard techniques and a specific anti-rat AM1-50 antibody. We confirmed the presence of a low AM-immunoreactivity in cardiomyocytes and cardiac fibroblasts, as well as in endothelial and smooth muscle cells of coronary vessels. Moreover, we provided evidence of the presence in both atria and ventricles of sparse voluminous AM-positive cells, mainly located near coronary vessels. These cells had the same juxtavascular location of toluidine blue-positive mast cells and their number decreased upon acute exposure to the 48/80 compound. However, ICC showed that in these cells AM was always colocalized with atrial and brain natriuretic peptides. Moreover, AM-storing cells were also positive to MyHC-Apla2, indicating that they share some phenotypic features with immature smooth muscle cells. The number of AM-storing cells underwent a dramatic decrease in response to the potent vasoconstrictor ET 1, suggesting an acute release of stored vasodilatory AM aimed at counteracting coronary constriction. Taken together, our present findings support the hypothesis that these cells may represent a novel subset of endocrine cells, strategically located near blood vessels in the mammalian heart, where they can release vasoactive peptides. PMID- 15754016 TI - Immunolocalization of platelet glycoprotein IIb/IIIa and P-selectin, and neutrophil-platelet interaction in human coronary unstable plaques. AB - Platelet aggregation at the site of plaque rupture or erosion is a dominant feature in the pathophysiology of plaque destabilization. To elucidate the role of glycoprotein (GP) IIb/IIIa in coronary plaque destabilization, we immunohistochemically studied the presence of GP IIb/IIIa in coronary atherectomy specimens obtained from patients with stable angina (SAP) and unstable angina pectoris (UAP). Moreover, we immunohistochemically investigated the presence of P selectin, which is known to be a marker of platelet activation, in these specimens. All these patients underwent atherectomy at primary atherosclerotic lesions responsible for SAP (n=25) and UAP (n=23). Frozen samples were studied with antibodies against smooth muscle cells, macrophages, neutrophils, endothelial cells, GP IIb/IIIa and P-selectin. Immunoreactive positive areas for GP IIb/IIIa, P-selectin, and macrophages, respectively, were calculated using computer-aided planimetry, and numbers of neutrophils were also counted. In the culprit lesions of UAP patients, 17 of the 23 lesions (74%) contained GP IIb/IIIa positive platelet thrombi, and all these platelet thrombi were positive for P selectin. In contrast, in the lesions of SAP patients, 3 of the 25 lesions (12%) showed staining positivity for GP IIb/IIIa and P-selectin. Quantitatively, the percentage of GP IIb/IIIa- and P-selectin-positive area was significantly higher (GP IIb/IIIa, P<0.0005; P-selectin, P<0.0001) in patients with UAP than in patients with SAP. The number of neutrophils was significantly higher (P<0.0005) in patients with UAP than in patients with SAP. Moreover, the percentage of GP IIb/IIIa-positive area showed a significant positive correlation with the number of neutrophils (r=0.66, p<0.0001). These findings strongly suggest that platelet activation and aggregation, leading to formation of platelet thrombi, and the interaction between activated platelets and neutrophils play a pivotal role in the pathogenesis of plaque destabilization in human coronary atherosclerotic lesions. PMID- 15754017 TI - Human adrenomedullin gene silencing by short interfering RNAs: a preliminary study. AB - Adrenomedullin (AM) is a regulatory peptide widely expressed, along its receptors, in cells and tissues, of which it controls many basic and specific functions acting in an autocrine-paracrine manner. However, the unequivocal demonstration of the physiological relevance of the regulatory role of AM would require the study of cells where endogenous AM system had been suppressed. Hence, we developed a protocol to silence the human AM gene by transfection with short interfering RNAs (siRNAs). Eight possible AM-siRNA sequences were designed: six siRNAs were synthesized in our laboratory and two were provided by Ambion. As positive control the suppression of the glyceraldehyde-3-phosphate dehydrogenase (GADPH) gene was tested using the Ambion Silencer GADPH siRNA kit. Cultured human embryonal kidney cell line HEK-293 and human umbilical vein endothelial cells (HUVECs) were transfected using either the Qiagen or the Ambion transfection reagent, and transfection visualization, carried out using Cy3-labeled siRNA and examining red fluorescence within the cells, showed that the former reagent was the most efficient. AM-gene silencing was determined in HUVECs by measuring AM mRNA levels in transfected and control cells by real-time polymerase chain reaction. Only Ambion siRNAs were effective, and the best AM-gene silencing (about 80%) was observed 48 or 72 h after transfection with 3 or 6 microg of siRNAs. The conclusion is drawn that siRNA technology can be useful in the investigations on AM functions, but that the complete suppression of the AM-gene transcription is very difficult to obtain. PMID- 15754018 TI - Down-regulation of DMBT1 gene expression in human oral squamous cell carcinoma. AB - Deleted in malignant brain tumors 1 (DMBT1) gene was recently isolated on chromosome 10q25.3-26.1 and has been proposed as a putative candidate tumor suppressor for brain, esophageal, gastric, colorectal, and lung cancer. However, little is known about the association of DMBT1 with oral squamous cell carcinoma (OSCC). To study the role of DMBT1 gene in OSCC oncogenesis, we examined 9 OSCC derived cell lines and 45 primary OSCC tissue specimens with respective normal tissues. Semi-quantitative reverse transcriptase chain reaction (RT-PCR) analysis revealed down-regulation or deletion of DMBT1 expression in all of the 9 cell lines and in 18 (40%) of 45 primary OSCC tissues. Additionally, 57 OSCC tissue specimens were examined by immunohistochemical staining of protein showing down regulation of DMBT1 protein in 31 (56.1%) of the 57 primary OSCC tissue specimens. To assess restoration of DMBT1 expression by demethylation of promoter region, the 9 cell lines were treated with 5-aza-2-deoxycytidine (5-Aza-C), one of the DNA demethylating agents. Six (66.7%) of 9 cell lines demonstrated restoration of DMBT1 expression after 5-Aza-C treatment. These results suggest that DMBT1 gene is involved in OSCC oncogenesis and/or progression and that methylation of promoter region is one of the important mechanisms suppressing the DMBT1 gene expression. PMID- 15754019 TI - Plasma level and tissue expression of angiogenic factors in patients with hereditary hemorrhagic telangiectasia. AB - The value of angiogenic factors interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF) was determined in patients with hereditary hemorrhagic telangiectasia (HHT) to evaluate their role in HHT pathogenesis. IL-8 and VEGF were measured in plasma of 41 HHT patients and healthy controls by ELISA technique. In both groups, the IL-8 and VEGF tissue expression in cryostat sections of the nasal mucosa were then compared. VEGF plasma levels were significantly increased in HHT patients compared to healthy controls. In contrast, the IL-8 plasma levels in both groups did not show any significant difference. Compared to healthy controls, HHT tissue samples showed a weak IL-8 staining, whereas the VEGF staining was very strong. The plasma levels of VEGF and IL-8 could not be correlated to the patients' clinicopathological findings. Additionally to the angiogenic pro-inflammatory cytokine IL-8, the angiogenic factor VEGF seems to play a major role in HHT pathogenesis. PMID- 15754020 TI - Status of reduced expression and hypermethylation of the APC tumor suppressor gene in human oral squamous cell carcinoma. AB - The adenomatous polyposis coli gene (APC gene) originally was identified as a tumor suppressor gene in colon cancer. We reported previously that APC is mutated and/or deleted in primary oral squamous cell carcinoma (OSCC) tissues and suggested that loss of APC function contributes to carcinogenesis in the oral region. In this study, we examined 50 OSCC tissue samples, which had been fixed in 10% buffered formaldehyde solution and embedded in paraffin, and eight cell lines, which were derived from OSCC, to analyze the expression level of the APC gene. Significant down-regulation of APC was detected by immunohistochemistry in 15 (30.0%) of 50 tissue samples and by the reverse transcriptase-polymerase chain reaction in five (62.5%) of eight cell lines. We then investigated the status of APC gene promoter methylation and restoration of the APC gene mRNA. Hypermethylation of the APC promoter CpG island was detected in two of eight (25%) OSCC-derived cell lines, and APC gene mRNA was restored in all OSCC-derived cell lines showing down-regulation of gene expression (n=5) after treatment with 5-aza-2'-deoxycytidine, a DNA demethylating agent. Thus, the contribution of down regulated APC expression to the development of human OSCC was about 30%, and hypermethylation of the gene promoter CpG island was confirmed to be a significant mechanism of inactivation of the APC gene in oral carcinogenesis. PMID- 15754021 TI - Protein and mRNA expression levels of VEGF-A and TGF-beta1 in different types of human coronary atherosclerotic lesions. AB - It is well documented that inflammation plays a major role in the establishment and progression of atherosclerosis. Endothelial cells, vascular smooth muscle cells and monocytes/macrophages are involved in this process by expressing inflammatory factors. The aim of the present study was to evaluate potential association and risk of VEGF-A and TGF-beta1 in human coronary atherosclerotic lesions. Twenty-six fresh human coronary artery segments were collected at autopsy. Conventional histology was performed and samples were classified into: no lesion group (NL), fatty streak group (FS), plaque group (P) and complicated lesion group (CL) based on the atherosclerotic lesion type. RNA extraction analysis with RT-PCR and immunohistochemistry was also performed. We observed that VEGF-A protein and mRNA expression increased during atherogenesis. The expression levels (protein and mRNA levels) of TGF-beta1 were decreased from NL to the FS group while, strong protein-staining and signal of mRNA expression in P and CL groups were observed. Our findings suggest a crucial role of VEGF-A in the development of coronary artery disease. The high protein and mRNA expression levels of TGF-beta1 in P and CL suggest that this factor may be implicated in the deposition of excessive extracellular matrix in the intima of the vessel wall, contributing to the expansion of the atheromatic plaque. PMID- 15754022 TI - Expression of the gene for hematopoietic cell specific protein is not restricted to cells of hematopoietic origin. AB - Hematopoietic cell line specific protein (HS1) is an intracellular signaling protein that has been reported as specifically expressed in hematopoietic cells. HS1 is known as a major substrate of protein-tyrosine kinases following activation by B-cell or T-cell receptor complexes. We report the first evidence that HS1 is also expressed in a variety of tissues different from hematopoietic tissues by using sensitive expression analysis including real-time quantitative RT-PCR. While former studies on HS1-expression were mostly limited to cells of hematopoietic origin, we screened a larger number of human tissues including tumor samples. Normal lung tissue showed a high degree of HS1 expression, second to the expression of hematopoietic cells. Expression of HS1 in tumor tissues was also clearly detectable. Our findings suggest that the signaling protein HS1 is involved in pathways different from the ones that have a specific role in the intracellular processes of hematopoietic cells. PMID- 15754023 TI - Amplification and functional characterization of MUC1 promoter and gene virotherapy via a targeting adenoviral vector expressing hSSTR2 gene in MUC1 positive Panc-1 pancreatic cancer cells in vitro. AB - Mucin1 (MUC1) promoter has been cloned from the 5' flanking region of the MUC1 gene in breast carcinoma, functionally characterized and applied in gene therapy of breast and esophageal carcinoma. In the present study, we amplified a 786 base pair (bp) MUC1 promoter by two-step nest PCR, and identified the activity and tumor-specificity using an enhanced green fluorescent protein (EGFP) gene as a reporter gene by fluorescence microscopy and flow cytometry analysis in Panc-1, primary normal pancreatic (PNPC), and cervical cancer HeLa cell lines. Subsequently, the human somatostatin receptor subtype 2 (hSSTR2) gene driven by MUC1 promoter was cloned into the pAdTrack to produce recombinant adenovirus AdMUC1-hSSTR2. The anticancer effect of AdMUC1-hSSTR2 was determined in Panc-1. The results demonstrated that there was no AdMUC1-hSSTR2-induced apoptosis, but a significant cell proliferation inhibition even without somatostatin (SST) analogue Octreotide, involved in the up-regulation of the cyclin-dependent kinase (CDK) inhibitors p21 and p27. Moreover, the anticancer effect could not be augmented by the addition of Octreotide, revealing a mechanism that was independent from induction of Octreotide. Therefore, this adenovirus system can be used as a novel, potent and specific tool for gene-targeting therapy in the MUC1 positive pancreatic carcinoma as shown in Panc-1. PMID- 15754024 TI - Polymorphisms and mutations of the folate receptor-alpha gene and risk of gastric cancer in a Chinese population. AB - Folate deficiency is associated with increased risk of many human diseases including several cancers. Folate receptor-alpha (FR-alpha) plays an important role in mediating influx of folates into cells and its expression can be induced by some environmental risk factors. Our previous study showed that decreased expression of FR-alpha occurs in human gastric cancer cell lines. In order to reveal the molecular mechanism of the difference of FR-alpha expression among these cell lines and analyze the relation between the mutations or/and polymorphisms of FR-alpha gene and gastric cancer, we screened the mutations of global FR-alpha gene in 12 gastric cancer cell lines and 28 gastric tumors and matched normal tissues by using PCR-DHPLC and DNA sequencing techniques, then further checked the exon2 of FR-alpha gene in 138 gastric tumors and matched normal tissues. Furthermore, we examined the relationship between two polymorphisms of FR-alpha A1314G and C1816delC and risk for gastric cancer in 296 cases and 354 age and sex-matched controls in northern China. We found that the allele frequency of FR-alpha1314A among cases was significantly higher than that among controls (0.186 vs. 0.143, P=0.014). Subjects with the A/G and A/A genotype had an increased risk for developing gastric cancer compared with the G/G genotype (OR=1.55; 95% CI=1.10-2.17). These results support the hypothesis that genetic polymorphism in the FR-alpha gene may contribute to susceptibility to carcinogenesis of the gastric cancer in the at-risk Chinese population. PMID- 15754025 TI - Extracellular matrix activity and caveolae events contribute to cell surface receptor activation that leads to MAP kinase activation in response to UV irradiation in cultured human keratinocytes. AB - Activation of cell surface components has been implicated in the activation of downstream signaling cascade in response to UV irradiation, and yet the identity and the interaction of those components have been scantly documented. Accumulating evidence indicates that caveolae encapsulating caveolins is the location for those interactions. We found in cultured human keratinocytes that UV irradiation induced both caveolin-1 and EGFR phosphorylation. Filipin, a caveolae disruptive agent, inhibited UV-induced caveolin-1 activation. Na+-K+-ATPase catalyzes active transport of Na+ and K+ across plasma membrane of mammalian cells, inactivation of which has recently been shown to be involved in the activation of signal transduction pathways including MAP kinase cascade. We found in this study that UV inactivated Na+-K+-ATPase in time-dependent manner, Na+-K+ ATPase activity started to decrease 5 min post UV irradiation and reduced to 60% of its original activity within 1 h. Pretreatment with Flipin and MMP inhibitor recovered Na+-K+-ATPase activity lost by UV irradiation. ECIS analysis indicated that both EGF treatment and UV irradiation increased membrane electric activity which was inhibited by MMP inhibitor and Filipin. Further study showed that pretreatment of human keratinocytes with MMP inhibitor or Filipin inhibited UV induced phosphorylation of p38 and JNK, which was however not observed in LnCap cells, a prostate cancer cell line lacking caveolin-1. UV irradiation also induced ectodomain shedding of HB-EGF in a time-dependent manner in keratinocytes. Collectively, we conclude that UV-induced MAP kinase activation is mediated by cell surface receptor activation due to the matrix activity and membrane caveolae function and inactivation of Na+-K+-ATPase. PMID- 15754026 TI - Modulation of the CXC-chemokine expression profile on tumor cells by the immunosuppressive drug mycophenolate mofetil. AB - The most undesirable complication of an effective immunosuppressive therapy is neoplastic tumor recurrence or the development of de novo cancer. Though the immunosuppressive drug, mycophenolate mofetil (MMF), has been introduced into clinical practice, no data dealing with the influence of MMF on tumor cell malignancy are available. We analyzed the adhesion capacity of colon, pancreas and kidney carcinoma cell lines to endothelium, as well as their chemokine profile before and after MMF treatment. Tumor cell adhesion to endothelial cell monolayers was evaluated in the presence of 0.1, 1, and 10 microM MMF and compared to unstimulated controls. Chemokine analysis concentrated on the CXC family, including 6 CXC-receptors (CXCR) and 15 CXC-ligands (CXCL), and was carried out by reverse transcriptase-polymerase chain reaction and flow cytometry. MMF strongly diminished the adhesion capacity of HT-29 colon tumor cells but not of DanG pancreas tumor cells to endothelium. MMF also had a strong impact on the chemokine profile of colon, kidney and pancreas carcinomas, whereby individual changes were observed, depending on the tumor type. Down-regulating effects on chemokines did not correlate with down-regulating effects on tumor cell adhesion. Since several of the chemokines investigated are regulatory elements in the process of cell transformation, dissemination and angiogenesis, we speculate that MMF might prevent post-transplant tumor recurrence and transendothelial migration. However, the efficacy of MMF might differ according to the tumor type. PMID- 15754027 TI - Heat shock protein 90 is required for increased DNA binding activity of activator protein-1, a heterodimer of Fos/JunD, in rheumatoid synovial cells under inflammatory stimuli. AB - We have studied the DNA binding profiles of activator protein-1 (AP-1) involved in synovial overgrowth and osteoporosis in rheumatoid arthritis (RA) in relation to the molecular chaperon heat shock protein 90 (HSP90). The AP-1 binding activity of the nuclear extracts of rheumatoid synovial cells was basically increased as compared with osteoarthritic synovial cells. Upon stimulation with inflammatory cytokines IL-1beta or TNFalpha, the AP-1 binding activity was further increased in rheumatoid synovial cells, and increased AP-1 protein was composed as heterodimers of Fos and JunD which was not known before as a major component of AP-1 in rheumatoid synovial cells. The increase of AP-1 binding activity as induced by inflammatory cytokines was specifically inhibited by geldanamycin, radicicol or herbimycin A, specific inhibitors of HSP90, while AP-1 protein was not decreased by geldanamycin. Further, HSP90 protein was not decreased by the inhibitors. The findings indicate that HSP90 is required for increased AP-1 binding activity of rheumatoid synovial cells under inflammatory stimuli and that AP-1 binding activity is inhibited by functionally inactivating HSP90 with the inhibitors. PMID- 15754028 TI - Clinical significance of abnormal lipoprotein patterns in liver diseases. AB - We analyzed lipids in liver diseases by agarose gel electrophoresis, and differential staining and simultaneous analysis of the cholesterol (Chol) and triglyceride (TG) fractions. Liver diseases were classified into chronic hepatitis (CH), liver cirrhosis (LC), hepatocellular carcinoma (HCC), and metastatic liver cancer, and each fraction was compared among these diseases. Atypical patterns that were unclassifiable according to the WHO classification of hyperlipidemia phenotypes were classified, and their clinical importance was evaluated. With progression of the pathologic conditions of CH, LC, and HCC, the T-Chol level, each Chol fraction, and the TG fraction decreased while the LDL-TG fraction increased. Metastatic liver cancer showed a lower HDL-fraction level but higher levels of the other parameters than HCC. When the subjects were classified into survivors and patients who died, the HDL fraction level in HCC and metastatic liver cancer, and the LDL level in LC and metastatic liver cancer differed between survivors and patients who died. Phenotypes of hyperlipidemia also differed among diseases, and atypical patterns were frequently observed in patients who died. There were 6 atypical patterns, of which 4 (slow alpha HDL, abnormal LDL, Lp-X, and Lp-Y) were associated with liver diseases. Slow alpha HDL appeared during slight bile stagnation and was accompanied by increases in the apo E level and the HDL particle size. Abnormal LDL appeared with severe liver dysfunction; a TG peak appeared at the position of LDL, and the HDL and VLDL fractions were negligible. Lp-X was a Chol-rich band, occurring on the cathode side of LDL in the presence of marked bile stagnation such as that in obstructive jaundice, and was accompanied by appearance of abnormal LDL. Lp-Y was similar to Lp-X in terms of mobility and associated diseases but contained Chol and TG. Abnormal LDL, Lp-X, and Lp-Y were often observed in patients with poor outcomes. Lipid analysis in liver diseases by this method showed results reflecting the pathologic conditions and may be clinically useful. PMID- 15754029 TI - Analysis of the apolipoprotein(a) size polymorphism in patients with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is characterized by an increased incidence of vascular disease which is only partially explained by traditional risk factors. Previous reports suggested that the level of lipoprotein(a) [Lp(a)], a particle linked to atherothrombotic disorders, is increased in patients with SLE. However, whether there are any differences in the distribution of apolipoprotein(a) [apo(a)] phenotypes between SLE patients and healthy controls remain to be determined. To address this issue, Lp(a) levels and apo(a) isoform size were analyzed in a total of 54 patients with SLE and in 108 age- and gender-matched healthy controls. SLE patients showed Lp(a) levels [median (interquartile range): 25.3 (6.5-51.0) vs. 9.5 (4.6-25.9) mg/dl, P=0.0109)] and a percentage of subjects with at least one small-sized apo(a) isoform (< or =25 K-IV repeats) significantly higher than controls (44.44% vs. 25.92%, P=0.0277). Multiple regression analysis adjusting for age, gender, disease duration, kidney involvement, the presence of active disease, as well as the carriage of at least one small apo(a) isoform revealed that only small apo(a) phenotypes were significant predictors of Lp(a) levels in SLE patients (P=0.0001). We conclude that genetic factors related to apo(a) size are a major determinant of elevated Lp(a) levels in patients with SLE. As small apo(a) phenotypes have been related to adverse vascular effects, it is feasible that small apo(a) isoforms may be a useful biological marker in the assessment of vascular risk in patients with SLE. PMID- 15754030 TI - Involvement of NF-kappaB and mitochondrial pathways in docetaxel-induced apoptosis of human oral squamous cell carcinoma. AB - Apoptosis induced by docetaxel that interferes with microtubule polymerization dynamics and is used clinically to treat advanced cancers, has not been fully defined in squamous cell carcinoma. In this study, apoptotic events involved in docetaxel treatment were investigated. When the human oral squamous cell carcinoma cell line HSC-3 was exposed to docetaxel for 72 h, a dose-dependent effect was observed in apoptosis using the TUNEL method. We observed activation of caspase cascade including activities like caspase-3, -8, and -9. And the pan caspase inhibitor z-VAD-fmk prevented apoptosis induced by docetaxel (0.1 microM), showing participation of caspases in this process. Since an antagonistic CD95-antibody (ZB4) exerted no effect on docetaxel-induced apoptosis, CD95/CD95L interaction was not involved in this pathway. The caspase-8-like activity was inhibited not only by IETD-fmk (caspase-8) but also by DEVD-fmk (caspase-3). The results indicate that the caspase-8-like activation occurred downstream of DEVDase. Docetaxel promoted the formation of reactive oxygen species (ROS) in mitochondria, and preincubation of cells with anti-oxidants such as N-acetyl cysteine and pyrrolidine dithiocarbamate, protected against apoptosis mediated by docetaxel. Furthermore, treatment with docetaxel elicited reduction of mitochondrial membrane potential, and release of cytochrome c to cytosol, after 48 h of treatment. We observed binding activity to NF-kappaB consensus site and interference with the mitochondrial function via NF-kappaB after docetaxel treatment. Preventing pro-apoptotic property of NF-kappaB inhibited docetaxel induced apoptosis. Thus, these results suggest that, following the activation of NF-kappaB by docetaxel, apoptosis is elicited through a mitochondria-dependent pathway. PMID- 15754032 TI - Expression of hTERT mRNA in a mortal liver cell line during S phase without detectable telomerase activity. AB - Normal human liver cells have a limited capacity for proliferation due to telomere shortening, whereas immortalized cells prevent shortening of the 3' single strand telomeric repeat by expressing telomerases, including human telomerase reverse transcriptase (hTERT). The hTERT transcript contains three deletion sites that give rise to alternatively spliced variants (ASVs). Recently, hTERT expression was observed in cycling primary presenescent human fibroblasts, which were believed to lack hTERT expression and telomerase activity. hTERT mRNA was expressed in the synthesis (S) phase of the cell cycle. Although hTERT mRNA has eight isoforms, it is not known which of the hTERT ASVs are expressed in S phase. In order to determine the possible relationships between the cell cycle and ASV expressions, we measured the full-length isoform and ASVs of hTERT mRNA in a mortal liver cell line and immortal cell lines that were synchronized in S phase of the cell cycle. Using RT nested-PCR analysis, the full-length isoform and alpha-deletion ASV of hTERT were detected in the LI90 mortal liver cell line at points when cells in S phase represented >48% of the cell population without detectable telomerase activity. hTERT was always expressed in the HLE and Huh-7 hepatocellular carcinoma cell lines, regardless of the cell cycle. Our results suggest the possibility that telomerase is regulated in a cell cycle-dependent manner in normal liver cells. PMID- 15754031 TI - beta-Cryptoxanthin stimulates cell proliferation and transcriptional activity in osteoblastic MC3T3-E1 cells. AB - The carotenoid beta-cryptoxanthin has been shown to have a stimulatory effect on bone formation in rat bone tissues in vitro. The effect of beta-cryptoxanthin in osteoblastic cells in vitro was investigated. Osteoblastic MC3T3-E1 cells were cultured for 72 h in alpha-minimal essential medium containing 10% fetal bovine sereum (FBS) to reach subconfluent monolayers. After culture, the medium was changed, then beta-cryptoxanthin (10(-8) to 10(-6) M) was added in the culture medium without FBS, and the cells were cultured for an additional 24, 48, or 72 h. The proliferation of osteoblastic cells was significantly enhanced in the presence of beta-cryptoxanthin (10(-8) to 10(-6) M), when it was cultured for 48 or 72 h in medium containing 10% FBS. When osteoblastic cells with subconfluency were cultured for 48 or 72 h in FBS free-medium containing beta-cryptoxanthin (10(-8) to 10(-6) M), alkaline phosphatase activity or deoxyribonucleic acid (DNA) content in the cells was significantly increased. Also, protein content in the cells was significantly increased by culture with 10(-6) M beta-cryptoxanthin for 48 or 72 h. The effect of beta-cryptoxanthin (10(-6) M) in increasing protein content, alkaline phosphatase activity, or DNA content in the cells was significantly blocked in the presence of staurosporine (10(-6) M) or PD98059 (10( 6) M), which is an inhibitor of protein kinases. The stimulatory effect of beta cryptoxanthin (10(-6) M) on cellular biochemical components was completely prevented in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; 10(-9) M), an inhibitor of transcriptional activity. The expressions of insulin-like growth factor (IGF)-I and transforming growth factor (TGF)-beta1 mRNAs were demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in osteoblastic cells using mouse IGF-I or TGF-beta1-specific primers. These expressions were significantly raised in the presence of beta-cryptoxanthin (10( 6) M). This study demonstrates that beta-cryptoxanthin has a stimulatory effect on cell proliferation and biochemical components in osteoclastic MC3T3-E1 cells, and that the carotenoid can stimulate transcriptional activity in the cells. PMID- 15754033 TI - Changes in gene expression level for defense system enzymes against oxidative stress and glutathione level in rat administered paraquat. AB - The herbicide paraquat (PQ) forms reactive oxygen species during enzymatic activation. We examined the effect of PQ on the relative levels of gene expression of antioxidant enzymes and glutathione (GSH) status in lungs of rats exposed to 20 mg/kg PQ. At 16 h after PQ intake, the mRNA expression level of glutathione reductase (GR) showed the greatest increase, and those of catalase (CAT) and manganese-superoxide dismutase (MnSOD) showed more modest increases. In contrast, PQ had little or no effect on the levels of mRNAs for copper/zinc superoxide dismutase (CuZnSOD) and glutathione peroxidase (GPX). These findings indicate that CAT and MnSOD are coordinated and play a major role in removal of oxidants. On the other hand, PQ caused a significant increase in the GSH level in the lungs, but not in the liver. This increase in the lungs was, at least in part, caused by stimulation of the gamma-glutamylcysteine synthetase gene. However, the expression of GPX mRNA was not stimulated as described above. Because GSH is a substrate for GPX and serves as a scavenger of hydroxyl radicals, the increase in GSH as well as GR expression may be insignificant. This imbalance may be a result of oxidative stress due to PQ. PMID- 15754034 TI - Effects of middle molecular weight fucoidans on in vitro and ex vivo angiogenesis of endothelial cells. AB - Native fucoidan, sulfated L-fucose polymer isolated from marine brown algae, has diverse biological activities including anticoagulant, anti-angiogenic and anti tumor effects. Its anti-angiogenic effect is of interest and the mechanisms have been studied. However, recently, proangiogenic effect of low molecular weight (LMW) fucoidans has been demonstrated. To clarify the opposite effects of fucoidan and LMW fucoidans, it is necessary to address the relationship between the molecular size and the effect on angiogenesis. In this study, the effects of middle molecular weight (MMW) fucoidans (15-30 kDa) on human umbilical vein endothelial cell functions were examined. Fucoidan (30 kDa) had similar effects to native fucoidan; inhibition of HUVEC tube formation and angiogenesis in an ex vivo model, although their effects were weaker than native fucoidan. On the other hand, 15-20 kDa fucoidan enhanced HUVEC migration, but did not inhibit HUVEC tube formation. Thus, 15-20 kDa fucoidan would have proangiogenic effect on angiogenesis. These results elucidate that 20-30 kDa would be a critical point to characterize the role of fucoidans on angiogenesis. PMID- 15754035 TI - Vitamin D in normal and pathological parathyroid glands: new prospects for treating hyperparathyroidism (review). AB - The secosteroid hormone active vitamin D [1,25(OH)2D3] is a key player in the regulation of calcium homeostasis and bone mineralization. In addition, it has antiproliferative and prodifferentiating effects on various cells in vitro and in vivo. The action of 1,25(OH)2D3 is mediated through the vitamin D receptor (VDR), which belongs to the superfamily of steroid/thyroid hormone nuclear receptors. VDR is expressed in the intestine, bone, kidney, parathyroid glands, and in many other tissues and cell types. In the parathyroid glands, 1,25(OH)2D3 markedly decreases parathyroid hormone gene transcription and parathyroid cell proliferation and induces parathyroid cell differentiation. Diminished VDR expression is frequent in parathyroid tumors and probably contributes to parathyroid tumorigenesis. The enzyme responsible for catalyzing synthesis of 1,25(OH)2D3 (1alpha-hydroxylase) has lately been demonstrated in the parathyroid glands. This indicates a new role for 1alpha-hydroxylase as an intracrine modulator of vitamin D function in non-renal tissues, which recently has been recognized as crucial in parathyroid tumor development. The growth-inhibitory properties of 1,25(OH)2D3 are prospects for treatment of hyperparathyroidism. PMID- 15754037 TI - Identification of active siRNAs against IGF-IR of porcine coronary smooth muscle cells in a heterologous cell line. AB - Testing of short interfering RNAs (siRNAs) to knock-down gene expression is complicated in primary differentiated cells due to their limited availability and short in vitro life span. The objective of this study was to bypass these limitations by testing selected siRNAs in a heterologous cell line. A plasmid containing a fragment of porcine IGF-I receptor (pIGF-IR) gene cloned downstream of EGFP sequence was constructed and cotransfected with pIGF-IR/siRNAs in HEK 293T human cell line. The abatement of EGFP fluorescence, measured at mRNA and protein level, was compared with that induced by a EGFP/siRNA. Among the three pIGF-IR/siRNAs tested, one was active against the hybrid reporter gene as EGFP/siRNA, while the other two were inactive. When transfected in primary porcine coronary smooth muscle cells (PC-SMCs) and tested by real-time PCR, immunocytofluorimetry and cell motility assay, pIGF-IR/siRNAs confirmed the results obtained in HEK 293T cells, thus establishing that the search of active siRNAs addressed to a gene of a cell and/or a species of interest can be done in a heterologous cell line. PMID- 15754038 TI - Identification and characterization of a novel human nephronectin gene in silico. AB - We describe the identification and characterization of a human epidermal growth factor (EGF)-like gene, nephronectin (NPNT), which encodes predicted proteins most homologous to the mouse nephronectin (Npnt). Human NPNT is located at the chromosomal position 4q25. Two transcript variants, NPNTa and NPNTb, are identified. Sequence comparison indicated that the human NPNT proteins share 88% amino acid identity with their mouse homologs. The in silico expression analysis showed the NPNT expression in a number of embryonic and adult tissues. NPNT was expressed in fetal ear, eye, heart, lung and kidney. In adults, the NPNT expression was widely found in lung, kidney, brain, uterus, placenta, thyroid gland and blood vessels. This gene is therefore implicated in embryonic development as well as maintenance of various adult tissues. PMID- 15754036 TI - Ghrelin, peptide YY and their receptors: gene expression in brain from subjects with and without Prader-Willi syndrome. AB - Ghrelin and peptide YY (PYY) are peptides generally produced by the gastrointestinal organs which are involved in appetite regulation via highly specialized centers in the brain. Abnormal plasma ghrelin and PYY levels compared with controls have been reported for subjects with Prader-Willi syndrome (PWS) which is characterized by infantile hypotonia, poor suck reflex and failure to thrive followed by hyperphagia and marked obesity in early childhood. We studied gene expression of ghrelin, peptide YY, and their receptors (i.e., GHS-R1a, GHS R1b, and NPY2R) in six different brain regions (frontal cortex, temporal cortex, visual cortex, pons, medulla, and hypothalamus) obtained from three subjects with PWS, two individuals with Angelman syndrome, and six controls to determine if expression of these genes is detectable in different regions of the brain in subjects with and without PWS. In general, expression of these genes using RT-PCR was detected in all subjects and no obvious differences were seen in their pattern of expression between subjects with or without PWS. Additional studies including quantitative gene expression measurements will be required to further evaluate the role of these genes in the eating disorder seen in PWS. PMID- 15754039 TI - Phosphorylated and hypoacetylated mutant p53 enhances cisplatin-induced apoptosis through caspase-9 pathway in the absence of transcriptional activation or translation. AB - It is not completely understood how certain epithelial cells harboring mutant p53 have better response to chemotherapy. We investigate the mechanism of cisplatin induced apoptosis in two resistant cell lines (parental TCCSUP and R273L mutant p53 transfectant) and two sensitive cell lines (V143A and N247I mutant p53 transfectants). Activation of caspase 9 was demonstrated by Western blotting, and specific inhibitor for caspase 9 could inhibit apoptosis. Inhibitors for caspases 1, 2, 6, and 8 had no effect on apoptosis. Transcriptional repression of Bcl-2 occurred during apoptosis and could be reversed by the treatment of histone deacetylase inhibitor trichostatin A (TSA). The expression of Noxa, p53 inducible ribonucleotide reductase subunit 2 (p53R2), and p53 inducible death domain (PIDD) gene were not elevated with treatment of cisplatin (CDDP). Surface trafficking of Fas or Fas-L was not observed. Ser15 of wild-type p53 and mutant p53 was phosphorylated in response to cisplatin. Acetylation of wild-type p53 increased, while acetylation of mutant p53 decreased during cisplatin treatment. Both transcriptional inhibitor actinomycin D and translational inhibitor cycloheximide did not inhibit apoptosis. These results indicated that phosphorylated and hypoacetylated mutant p53 could enhance cisplatin-induced apoptosis through activation of caspase 9 independent of transcriptional activation and translation. PMID- 15754040 TI - Evaluation of proteasome-inhibitory and apoptosis-inducing potencies of novel (-) EGCG analogs and their prodrugs. AB - The anti-cancer and cancer-preventive effects of green tea and its main constituent (-)-epigallocatechin gallate [(-)-EGCG] are well documented by a variety of studies, including epidemiological, cell culture, animal, and clinical studies. While (-)-EGCG remains the most potent polyphenol in green tea, it is very unstable in neutral or alkaline conditions (i.e. physiologic pH). In an effort to discover more stable polyphenol proteasome inhibitors, we synthesized several novel (-)-EGCG analogs with -OH groups eliminated from the B- and/or D rings. In addition, we also synthesized their putative prodrugs with -OH groups protected by peracetate that can be removed by cellular cytosolic esterases. We first examined the structure-activity relationship of these unprotected and protected compounds to their proteasome-inhibitory potentials. We found that decreasing -OH groups from either the B- or D-ring leads to diminished proteasome inhibitory activity in vitro. However, in cultured tumor cells only the protected analogs were capable of potently inhibiting the proteasome activity. Furthermore, these protected analogs induced apoptotic cell death in a tumor cell-specific manner. The superior efficacy of the protected (-)-EGCG analogs indicates the formation of an entirely new compound(s) in intact tumor cells. These data suggest that the B-ring/D-ring peracetate-protected EGCG analogs have great potential to be developed into novel anti-cancer and cancer-preventive agents. PMID- 15754041 TI - Comparative genomics on Wnt3-Wnt9b gene cluster. AB - WNT signals, transduced through Frizzled (FZD) receptors with extracellular WNT binding domain and cytoplasmic Dishevelled-binding domain, are implicated in carcinogenesis and embryogenesis. WNT3-WNT9B (WNT14B) locus (17q21.31) and WNT3A WNT9A (WNT14) locus (1q42.13) are paralogous regions within the human genome. Here, the rat Wnt3 and Wnt9b genes were identified and characterized by using bioinformatics. Wnt3 and Wnt9b genes at rat chromosome 10q32.1 were clustered in head-to-head manner with an interval of about 24 kb within AC105632.3 genome sequence. The rat Wnt3 gene, consisting of five exons, encoded a 355-aa protein with N-terminal signal peptide, 24 conserved Cys residues and two Asn-linked glycosylation sites. The rat Wnt9b gene, consisting of four exons, encoded a 359 aa protein with N-terminal signal peptide, 24 conserved Cys residues and one Asn linked glycosylation site. The rat Wnt3 core promoter showed 80.5% nucleotide identity with human WNT3 core promoter, while rat Wnt9b core promoter showed 45.6% nucleotide identity with human WNT9B core promoter. MYB (c-Myb), ELK1, POU2F1 (OCT1), HNF4A (HNF-4), COMP1, NFYA (NF-Y) and NKX2-5 binding sites were conserved between rat Wnt3 and human WNT3 core promoters. The Wnt3-Wnt9b intergenic conserved region (IGCR), corresponding to nucleotide position 124747 125252 of AC105632.3 genome sequence, showed 85.6% nucleotide identity with human WNT3-WNT9B IGCR. GC content of rat Wnt3-Wnt9b IGCR was 59.5%. Wnt3-Wnt9b IGCR was predicted as regulatory element rather than gene because cDNA or EST derived from Wnt3-Wnt9b IGCR was not identified. This is the first report on the rat Wnt3 and Wnt9b genes as well as on comparative genomics on the Wnt3-Wnt9b gene cluster. PMID- 15754042 TI - Comparative genomics on Wnt5a and Wnt5b genes. AB - Canonical WNTs (WNT2, WNT2B, etc) activate the beta-catenin-TCF pathway to induce carcinogenesis, while non-canonical WNTs (WNT5A, WNT11, etc) activate the planar cell polarity (PCP) pathway to induce cell motility and metastasis. WNT5A gene at chromosome 3p14.3 and WNT5B gene at chromosome 12p13.33 are paralogs within the human genome. Here, we identified and characterized rat Wnt5a and Wnt5b genes by using bioinformatics. Rat Wnt5a and Wnt5b genes, consisting of five exons, were identified within AC095764.5 and AC112027.3 genome sequences, respectively. Rat Wnt5a (380 aa) and Wnt5b (359 aa) were secreted proteins with 24 conserved Cys residues and four Asn-linked glycosylation sites, which showed 75.8% total-amino acid identity. Nucleotide position 182586-183836 of AC095764.5 genome sequence and nucleotide position 161044-159886 of AC121764.2 genome sequence were identified as evolutionarily conserved rat Wnt5a and human WNT5A promoters, respectively. Nucleotide identity between rat Wnt5a and human WNT5A promoters was 72.5%. E47 and NKX2-5-binding sites were evolutionarily conserved among rat Wnt5a, mouse Wnt5a, and human WNT5A promoters. On the other hand, rodent Wnt5b promoters and human WNT5B promoter were significantly divergent. Up-regulation of Wnt5b during rodent adipocytic differentiation does not simply indicate the implication of WNT5B in human adipogenesis. Real susceptibility gene for type 2 diabetes, associated with SNP within intron 3 of human WNT5B gene (IMS JST024404), remains to be identified. This is the first report on rat Wnt5a and Wnt5b genes as well as on comparative genomics for Wnt5a and Wnt5b orthologs. PMID- 15754043 TI - Current pancreatogastrointestinal anastomotic methods: results of a Japanese survey of 3109 patients. AB - BACKGROUND: Pancreatogastrointestinal anastomosis is the most important anastomotic method of reconstructing the digestive tract following pancreaticoduodenectomy. METHODS: We therefore conducted a survey on pancreatogastrointestinal anastomosis at the 28th Japan Pancreatic Surgery Club in 2001. RESULTS: Results revealed that of the methods of pancreatogastrointestinal anastomosis, pancreatojejunostomy (82.9%) was overwhelmingly more popular than pancreatogastrostomy (17.1%). Anastomotic leakage occurred in 12.8% of cases, and 13.0% of these patients died. The incidence of anastomotic leakage was 11.0% with pancreatogastrostomy, and 13.3% with pancreatojejunostomy. There was no significant difference in the development of anastomotic leakage among the anastomotic organ used with the pancreas, the pancreas resection method, and the pancreatic juice drainage method. Investigation of intra-abdominal hemorrhage and abscess, which are serious complications that result from anastomotic leakage, revealed that intra-abdominal hemorrhage occurred in 1.1% of all cases, and intra-abdominal abscess was seen in 3.3% of all cases. However, there were no significant differences between pancreatogastrostomy and pancreatojejunostomy with respect to incidence rates of intra-abdominal hemorrhage and abscess or mortality. CONCLUSIONS: The method of pancreatogastrointestinal anastomosis should be chosen according to each individual case. PMID- 15754044 TI - Telomerase activity and Bcl-2 expression in gallbladders of pancreaticobiliary maljunction patients: a preliminary study. AB - BACKGROUND: The congenital anomaly pancreaticobiliary maljunction (PBM) is considered to be a precancerous disease. PBM carcinogenesis is believed to be an accumulation of gene abnormalities, but the early events causing PBM carcinogenesis are still unclear. In the present study, telomerase activity and Bcl-2 expression in the gallbladder mucosa of PBM and non-PBM gallbladders were investigated. METHODS: The operative gallbladder materials were from five control cases, two cases of non-PBM gallbladder cancer, three of PBM gallbladder cancer, and three of non-neoplastic PBMs. Multiple sampling was performed from each gallbladder. The studies performed were: (1) immunohistochemistry of p53, Ki-67, and Bcl-2; (2) survey of k-ras point mutations; and (3) measurement of telomerase activity in each sample. RESULTS: In the cases of non-PBM cancer, abnormalities from the above studies were detected only in the cancerous lesions. Normal appearing mucosa did not show Bcl-2 expression or telomerase activity. However, in the cases of PBM cancer, normal-appearing mucosa showed telomerase activity and Bcl-2 expression, but did not show p53, Ki-67, or k-ras abnormalities. In the non-neoplastic PBM, all samples showed Bcl-2 expression, and many showed telomerase activity. CONCLUSIONS: Bcl-2 expression and activation of telomerase are probably early events causing carcinogenesis of the PBM gallbladder mucosa. They might be important factors causing carcinogenesis associated with chronic inflammation. PMID- 15754045 TI - Should laparoscopic cholecystectomy be performed in patients with thick-walled gallbladder? AB - BACKGROUND/PURPOSE: Laparoscopic cholecystectomy is the procedure of choice for patients with symptomatic cholelithiasis. This procedure is contraindicated in patients with gall-bladder cancer (GBC) because of fear of dissemination of the disease. One of the findings raising the suspicion of GBC is a thick-walled gallbladder (TWGB). METHODS: A prospective study of patients with TWGB was done over a period of 10 months at a tertiary-level referral hospital in northern India. We studied the clinical profiles, investigations (ultrasound [US] and computerized tomography [CT]) and management plans in these patients. RESULTS: A total of 60 patients were included in the study. After cholecystectomy, histopathology of gallbladders showed GBC in 2 (3.3%) patients. The remaining 58 patients had chronic cholecystitis, of whom 28 (48%) had xanthogranulomatous variant chronic cholecystitis. Cholecystectomy by the laparoscopic method was attempted in 46 (77%) patients and by open technique in the remaining 14 (23%) patients. Laparoscopic cholecystectomy was successful in 40 of the 46 (87%) patients in whom it was attempted. Obscure anatomy, suspicion of GBC, and bile duct injury were the causes of conversion, in the remaining 13% (6/46). None of the 11 patients who had a CT examination because of clinical or US suspicion of malignancy turned out to have GBC at final histology. Both the cases of GBC in this study were incidental findings on final histopathology. CONCLUSIONS: Laparoscopic cholecystectomy can be successfully performed in the majority of patients with diffuse TWGB, with appropriate selection. There is, however, an increased chance of conversion to open cholecystectomy in these patients. If there is an intraoperative suspicion of GBC, early conversion to open cholecystectomy and frozen section/imprint cytology will help to decide the further treatment during surgery. PMID- 15754046 TI - Indications for pancreatoduodenectomy in patients undergoing lymphadenectomy for advanced gallbladder carcinoma. AB - BACKGROUND/PURPOSE: Carcinoma of the gallbladder shows diverse patterns of spread. The most appropriate surgical procedures according to the depth and extent of the spread of the tumor are still controversial. METHODS: We investigated this surgical problem clinicopathologically, especially regarding the indications for pancreatoduodenectomy (PD), by retrospectively reviewing the clinical records of 216 patients who were surgically treated for advanced gallbladder carcinoma. Detailed studies of resected specimens, in our department, showed that preservation of the pancreas head carried an increased risk of residual microscopic metastases in small peripancreatic lymphatic nodes and ducts. Some patients with curative operations had shown recurrence in lymph nodes around the head of the pancreas within a few years after PD. Therefore, we performed hepatopancreato duodenectomy (HPD) in 93 patients, some of whom underwent the HPD as a prophylactic dissection of peripancreatic lymph nodes and some of whom underwent the HPD as a curative resection due to invasion to the peripancreatic lymph nodes, duodenum, and pancreas, observed macroscopically. The surgical outcome after PD was compared with that obtained after other curative resections (non-HPD). RESULTS: In patients with microscopically negative lymph node metastasis without hepatoduodenal ligament invasion, PD was not necessary for a complete resection of lymphatic metastases around the pancreas head. The 5 year survival rate of these patients who had HPD was not significantly higher than that for non-HPD resections with curative intent (73% vs 63%), and lymphatic recurrence was not marked. On the other hand, in patients with positive lymph node metastases without hepatoduodenal ligament invasion, PD was necessary for a complete resection. The 5-year survival rate after HPD in these patients was significantly improved compared to that after non-HPD resections with curative intent (87% vs 17%), because lymphatic recurrence was reduced (0% vs 80%). CONCLUSIONS: In advanced carcinomas with positive hepatoduodenal ligament invasion, although the basic operative strategy, which inevitably includes right lobectomy, should be curative, PD should not be performed for prophylactic lymphatic resection, because local recurrence in the hepatoduodenal ligament cannot be controlled by PD, and the 5-year survival rate of curative HPD was only 4% (non-HPD resection with curative intent, 18%); moreover, frequent, lethal, major postoperative complications (hepatic failure) occurred after combined right lobectomy. PMID- 15754047 TI - Value of magnetic resonance cholangiopancreatography with secretin stimulation in the evaluation of pancreatic exocrine function after pancreaticogastrostomy. AB - BACKGROUND/PURPOSE: The aim of this study was to assess the value of magnetic resonance cholangiopancreatography with secretin stimulation (secretin-MRCP) in evaluating the remnant pancreatic exocrine reserve after pancreaticogastrostomy with pancreaticoduodenectomy. METHODS: Forty-three patients who had undergone pancreaticoduodenectomies and who were given pancreaticogastrostomies for reconstruction were studied. Dynamic MRCPs, using a half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequence were obtained before and up to 10 min after secretin administration. The morphologic features and diameter of the main pancreatic duct were monitored and graded before and after secretin stimulation. The results were compared with those of endoscopic findings, secretin stimulation testing with a collection of pancreatic fluid, N-benzoyl- L tyrosyl-p-aminobenzoic acid (BT-PABA) excretion testing, and fecal chymotrypsin concentration. RESULTS: The results of secretin-MRCP were classified into three distinct groups: a good-secretion group (group 1; n=22; 51%), a moderate secretion group (group 2; n=10; 23%), and a poor-secretion group (group 3; n=11; 26%). This MRCP classification correlated significantly with the concentrations of the pancreatic enzymes p-type amylase, lipase, and trypsin in the gastric juice. The BT-PABA test value was 59.8% in group 1, 46.1% in group 2, and 46.5% in group 3, and was significantly higher in group 1 than in groups 2 or 3. The fecal chymotrypsin concentration was 20.5 U/g in group 1, 14.5 U/g in group 2, and 0.7 U/g in group 3, and there was a significant correlation between the MRCP classification and fecal chymotrypsin concentration. CONCLUSIONS: MRCP with secretin stimulation favorably reflected the presence of remnant pancreatic exocrine function. Therefore, secretin-MRCP is a feasible and effective follow-up examination method to evaluate remnant pancreatic exocrine function after pancreaticogastrostomy. PMID- 15754048 TI - Epidermoid cyst in an intrapancreatic accessory spleen: a case report. AB - Ectopic splenic tissue in the abdominal cavity is a common entity, with a reported incidence of 10% in the general population. However, an intrapancreatic accessory spleen is a rare disease, and moreover cyst formation in it is exceedingly rare. A 58-year-old woman with a 25-mm multilocular cyst in the tail of the pancreas detected incidentally by ultrasonography was admitted for further evaluation. Because malignancy could not be ruled out, a spleen-preserving distal pancreatectomy was performed. The cut surface of the surgical specimen showed a multilocular cyst surrounded by brown solid tissue resembling normal spleen. Pathological examination revealed it was stratified squamous epithelium and was surrounded by splenic tissue. The final pathological diagnosis was epidermoid cyst in an accessory spleen in the pancreas. This cyst has no characteristic features on diagnostic imaging. Consequently, it is not possible to make a definite preoperative diagnosis in most cases. Epidermoid cyst in intrapancreatic splenic tissue is another lesion to be considered in the differential diagnosis of pancreatic tail tumors. PMID- 15754049 TI - A case of benign bile duct stricture causing difficulty in differential diagnosis from bile duct carcinoma. AB - We report a patient with benign bile duct stricture causing difficulty in differential diagnosis from bile duct carcinoma. A 66-year-old woman consulted a local physician because of general fatigue. Blood biochemical tests showed increased levels of biliary tract enzymes. Abdominal ultrasonography (US) revealed tapering and blockage of the midportion of the bile duct and dilation of the intrahepatic bile ducts. Magnetic resonance cholangiopancreatography (MRCP) demonstrated obstruction of the midportion of the bile duct. Later, because a marked increase in biliary tract enzymes and jaundice appeared, percutaneous transhepatic biliary drainage (PTBD) was performed. Post-PTBD cytological examination of bile was negative for cancer. A third biopsy showed slight hyperplasia with no malignant findings. Recholangiography, performed through PTBD, suggested gradual improvement of bile duct stricture, but could not completely exclude the possibility of malignancy; thus, resection of the bile duct including the stricture site was performed, and the resected specimen was submitted for intraoperative frozen section examination. Histopathological diagnosis did not reveal malignant findings. After cholecystectomy and bile duct resection, hepaticojejunostomy (Roux-en-Y) was performed. Because only erosion and desquamation of the mucosal epithelium and mild submucosal inflammatory cell infiltration and fibrosis were observed, chronic cholangitis was diagnosed histopathologically. Surgical resection of the bile duct should be considered for potentially malignant stricture of the bile duct. PMID- 15754050 TI - Excision of a juxtapapillary duodenal diverticulum causing biliary obstruction: report of three cases. AB - We report three cases of biliary obstruction caused by a juxtapapillary duodenal diverticulum that were treated by excision of the diverticulum. A 72-year-old man, a 77-year-old woman, and an 81-year-old woman each presented with recurrent obstructive jaundice. Diagnostic imaging revealed a juxtapapillary duodenal diverticulum compressing the common bile duct (CBD). Following cholecystectomy, the diverticulum between the intrapancreatic CBD and pancreatic parenchyma was isolated and excised successfully in each case. The patients have been followed up for 34, 31, and 22 months, respectively. In one patient, choledocholithiasis developed 33 months after the surgery, necessitating endoscopic sphincterotomy. Duodenal diverticulectomy is a useful procedure to relieve biliary obstruction caused by a juxtapapillary duodenal diverticulum. However, it remains unclear whether excision of the diverticulum is preferred to biliodigestive anastomosis from the point of view of long-term prognosis. Subsequent surveillance is necessary. PMID- 15754051 TI - Huge endocrine tumor of the pancreas. AB - We report a case of a huge endocrine tumor of the pancreas. A 76-year-old woman was admitted for a left upper abdominal mass. She showed neither hormonal symptoms nor abnormal serum hormone levels. Ultrasound imaging showed that a heterogeneously solid and partly calcified tumor was located in the retroperitoneal space. Computed tomography showed a hypervascular tumor with central degeneration. Open laparotomy revealed that the tumor originated from the pancreas; it was removed in a mass including the spleen and the pancreatic body and tail. The resected specimen demonstrated that the 580-g tumor was multi lobulated, encapsulated, and 14 x 14 x 7 cm in size. Microscopically, the tumor consisted of cuboidal cells with round nuclei; the cells formed palisade and trabecular structures. Immunohistologically, chromogranin A, neuronspecific enolase, and glucagon were positive only for the tumor cells. Final diagnosis was endocrine tumor of the pancreas immunohistochemically producing glucagon. No tumor recurrence has been found 3 years after the surgery. We conclude that huge tumors in the retroperitoneal space must be differentiated from nonfunctioning endocrine tumors of the pancreas because the clinical course is markedly different. PMID- 15754052 TI - Secondary structural effects on protein NMR chemical shifts. AB - For an amino acid in protein, its chemical shift, delta(phi, psi)(s), is expressed as a function of its backbone torsion angles (phi and psi) and secondary state (s): delta(phi, psi)(s=deltaphi, psi)_coil+Deltadelta(phi, psi)_s), where delta(phi, psi)(coil) represents its chemical shift at coil state (s=coil); Delta delta(phi, psi)(s) (s=sheet or helix) is herein defined as secondary structural effect correction factor, which are quantitatively determined from Residue-specific Secondary Structure Shielding Surface (RSS) for (13)CO, (13)Calpha, (13)Cbeta,(1)Halpha, (15)N, and (1)HN nuclei. The secondary structural effect correction factors defined in this study differ from those in earlier investigations by separating out the backbone conformational effects. As a consequence, their magnitudes are significantly smaller than those earlier reported. The present Delta delta(phi, psi)(sheet) and Delta delta(phi, psi)(helix) were found varying little with backbone conformation and the 20 amino acids, specifically for (13)CO, (13)Calpha, and (1)Halpha nuclei. This study also carries out some useful investigations on other chemical shift prediction approaches - the traditional shielding surfaces, SHIFTS, SHIFTX, PROSHIFT, and identifies some unexpected shortcomings with these methods. It provides some useful insights into understanding protein chemical shifts and suggests a new route to improving chemical shifts prediction. The RSS surfaces were incorporated into the program PRSI. PMID- 15754054 TI - Biosynthetic site-specific (13) C labeling of the light-harvesting 2 protein complex: a model for solid state NMR structure determination of transmembrane proteins. AB - Partly biosynthetic site-directed isotopically (13)C enriched photosynthetic light-harvesting 2(LH2) complexes have been prepared from Rhodopseudomonas acidophila strain 10050 by using chemically labeled [1,2,3,4-(13)C], [1,4-(13)C] and [2,3-(13)C] succinic acid as a precursor in the growth medium. Two dimensional proton driven spin diffusion (PDSD) solid state NMR correlation spectroscopy has been used to trace each individual (13)C isotope from the labeled succinic acid precursor to its destination into the protein and into the embedded major light-absorbing bacteriochlorophyll cofactors. For both the residues of the protein and for the cofactors distinct labeling patterns have been deduced, for protein complexes prepared from [1,4-(13)C]-succinic acid or [2,3-(13)C]-succinic labeled media. All residues, except isoleucine and leucine, have been labeled almost homogeneously by the succinic acid precursor. Carbonyl carbons in the protein backbone were labeled by [1,4-(13)C]-succinic acid, while the Calpha and Cbeta carbons of the residues were labeled by [2,3 (13)C]-succinic acid. Leucine and isoleucine residues were labeled using a uniformly labeled amino acid mixture in the medium. The pattern labeling yields an increase of the resolution and less spectral crowding. The partial labeling technique in combination with conventional solid state NMR methods at ultra high magnetic fields provides an attractive route to resolve chemical shifts for alpha-helical transmembrane protein structures. PMID- 15754053 TI - Probing membrane protein orientation and structure using fast magic-angle spinning solid-state NMR. AB - One and two-dimensional solid-state NMR experiments are discussed that permit probing local structure and overall molecular conformation of membrane-embedded polypeptides under Magic Angle Spinning. The functional dependence of a series of anisotropic recoupling schemes is analyzed using theoretical and numerical methods. These studies lead to the construction of a set of polarization dephasing or transfer units that probe local backbone conformation and overall molecular orientation within the same NMR experiment. Experimental results are shown for a randomly oriented peptide and for two model membrane-peptides reconstituted into lipid bilayers and oriented on polymer films according to a method proposed by Bechinger et al. PMID- 15754055 TI - Efficient assignment of methyl resonances: enhanced sensitivity by gradient selection in a DE-MQ-(H)CC(m)Ht (m)-TOCSY experiment. AB - We present a gradient selected and doubly sensitivity-enhanced DE-MQ-(H)CC(m)H(m) TOCSY experiment for the sequence-specific assignment of methyl resonances in (13)C,(15)N labeled proteins. The proposed experiment provides improved sensitivity and artifact suppression relative to the phase-cycled experiments. One part of the (13)Cchemical shift evolution takes place under heteronuclear multiple quantum coherence, whereas the other part occurs under (13)C single quantum coherence in a semi-constant time fashion. The feasibility of the experiment was assessed using (15)N,(13)C labeled Mus musculus coactosin (16 kDa), having a rotational correlation time of 14.5 ns at 15 degrees C in D(2)O. A 16-h experiment on 600 MHz (1)H yielded good quality data and enabled the assignment of 70 out of 72 methyl groups in coactosin. As well as being an improved approach for methyl resonance assignment, this experiment can also be highly valuable for the rapid assignment of methyl resonances in SAR by NMR studies. PMID- 15754056 TI - Adiabatic-passage cross polarization in N-15 NMR spectroscopy of peptides weakly associated to phospholipids: determination of large RDC. AB - Structural information can be extracted from one-bond residual dipolar couplings (RDC) measured in NMR spectra of systems in field-ordered media. RDC can be on the order of J-couplings if the anisotropy of alignment is approximately 10(-2), 10-fold stronger than that typically used for structural studies of water-soluble proteins. In such systems the performance of (1)H--> (15)N polarization transfer methods of the INEPT type is not satisfactory. In this study we show the effectiveness of adiabatic-passage cross-polarization (APCP) in transferring the (1)H--> (15)N polarization in the bicelle-associated peptide Leucine Enkephalin (Lenk). APCP is efficient both in static samples and in samples spun at the magic angle (MAS) or any other angle of the spinning axis to the magnetic field (variable-angle spinning, VAS). The anisotropic spectrum of an aligned static sample and the isotropic spectrum of the sample under MAS provide a set of possible values for the (1)H-(15)N RDC of phospholipid-associated Lenk. The unambiguous determination of the (1)H-(15)N RDC was accomplished by means of VAS experiments. PMID- 15754057 TI - Efficient production of isotopically labeled proteins by cell-free synthesis: a practical protocol. AB - We provide detailed descriptions of our refined protocols for the cell-free production of labeled protein samples for NMR spectroscopy. These methods are efficient and overcome two critical problems associated with the use of conventional Escherichia coli extract systems. Endogenous amino acids normally present in E. coli S30 extracts dilute the added labeled amino acids and degrade the quality of NMR spectra of the target protein. This problem was solved by altering the protocol used in preparing the S30 extract so as to minimize the content of endogenous amino acids. The second problem encountered in conventional E. coli cell-free protein production is non-uniformity in the N-terminus of the target protein, which can complicate the NMR spectra. This problem was solved by adding a DNA sequence to the construct that codes for a cleavable N-terminal peptide tag. Addition of the tag serves to increase the yield of the protein as well as to ensure a homogeneous protein product following tag cleavage. We illustrate the method by describing its stepwise application to the production of calmodulin samples with different stable isotope labeling patterns for NMR analysis. PMID- 15754058 TI - Asparagine and glutamine side-chain conformation in solution and crystal: a comparison for hen egg-white lysozyme using residual dipolar couplings. AB - Experimental (15)N-(1)H and (1)H-(1)H residual dipolar couplings (RDCs) for the asparagine (Asn) and glutamine (Gln) side chains of hen egg-white lysozyme are measured and analysed in conjunction with (1)N relaxation data, information about chi(1) torsion angles in solution and molecular dynamics simulations. The RDCs are compared to values predicted from 16 high-resolution crystal structures. Two distinct groups of Asn and Gln side chains are identified. The first contains residues whose side chains show a fixed, relatively rigid, conformation in solution. For these residues there is good agreement between the experimental and predicted RDCs. This agreement improves when the experimental order parameter, S, is included in the calculation of the RDCs from the crystal structures. The comparison of the experimental RDCs with values calculated from the X-ray structures shows that the similarity between the oxygen and nitrogen electron densities is a limitation to the correct assignment of the Asn and Gln side-chain orientation in X-ray structures. In the majority of X-ray structures a 180 degrees rotation about chi(2) or chi(3), leading to the swapping of N(delta/epsilon 2) and O(delta/epsilon 1), is necessary for at least one Asn or Gln residue in order to achieve good agreement between experimental and predicted RDCs. The second group contains residues whose side chains do not adopt a single, well-defined, conformation in solution. These residues do not show a correlation between the experimental and predicted RDCs. In many cases the family of crystal structures shows a range of orientations for these side chains, but in others the crystal structures show a well-defined side-chain position. In the latter case, this is found to arise from crystallographic contacts and does not represent the behaviour of the side chain in solution. PMID- 15754060 TI - (1)H, (15)N, and (13)C chemical shift assignments of the resuscitation promoting factor domain of Rv1009 from Mycobacterium tuberculosis. PMID- 15754059 TI - NMR structure of human coactosin-like protein. PMID- 15754061 TI - Assignment of the (1)H, (15)N and (13)C resonances of SufA from Escherichia coli involved in Fe-S cluster biosynthesis. PMID- 15754063 TI - (1)H, (13)C and (15)N sequence-specific resonance assignments of the two-domain thrombin inhibitor dipetalin. PMID- 15754062 TI - (1)H, (13)C, and (15)N resonance assignments and secondary structure of human pancreatitis-associated protein (hPAP). PMID- 15754064 TI - (1)H, (15)N and (13)C resonance assignment of human gammaS-crystallin, a 21 kDa eye-lens protein. PMID- 15754065 TI - (1)H, (13)C, and (15)N resonance assignments of the EscJ protein, a structural component of the Type III secretion system of enteropathogenic E. coli (EPEC). PMID- 15754066 TI - Economic evidence in affective disorders: a review. PMID- 15754067 TI - Economic evidence in anxiety disorders: a review. PMID- 15754068 TI - Economic evidence in brain tumour: a review. PMID- 15754070 TI - Economic evidence in dementia: a review. PMID- 15754071 TI - Economic evidence in epilepsy: a review. PMID- 15754072 TI - A review of European studies on the economic burden of brain diseases. PMID- 15754073 TI - Economic evidence in migraine and other headaches: a review. PMID- 15754074 TI - Economic evidence in addiction: a review. PMID- 15754075 TI - Economic evidence in multiple sclerosis: a review. PMID- 15754076 TI - Economic evidence in Parkinson's disease: a review. PMID- 15754077 TI - Economic evidence in psychotic disorders: a review. PMID- 15754078 TI - Economic evidence in stroke: a review. PMID- 15754079 TI - Economic evidence in trauma: a review. PMID- 15754081 TI - Bone mass and mineral metabolism in HIV+ postmenopausal women. AB - The objective of this cross-sectional study was to estimate the prevalence of and risk factors for osteoporosis in HIV+ postmenopausal women. Bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA) and biochemical indices of mineral metabolism were measured in 31 Hispanic and African American HIV+ postmenopausal women. BMD was compared with 186 historical controls, matched for age, ethnicity and postmenopausal status. Mean BMD was significantly lower at the lumbar spine and total hip in the HIV+ group, as compared with controls. Prevalence of osteoporosis was higher in the HIV+ group than controls at the lumbar spine (42% vs 23%, p =0.03) and total hip (10% vs 1%, p =0.003). Among HIV+ women, time since menopause and weight were significant predictors of BMD, while duration or class of antiretroviral therapy (ART), AIDS diagnosis, nadir CD4, steroid use, and vitamin D deficiency were not. Prevalence of osteoporosis is substantially higher in HIV+ Hispanic and African-American postmenopausal women than in controls. Established osteoporosis risk factors were more important in predicting BMD than factors associated with HIV infection and ART. Long-term management of the growing female HIV population should include the evaluation for and management of osteoporosis. PMID- 15754083 TI - Experimental compaction of anisotropic granular media. AB - We report on experiments to measure the temporal and spatial evolution of packing arrangements of anisotropic and weakly confined granular material, using high resolution gamma-ray adsorption. In these experiments, the particle configurations start from an initially disordered, low-packing-fraction state and under vertical solicitations evolve to a dense state. We find that the packing fraction evolution is slowed by the grain anisotropy but, as for spherically shaped grains, can be well fitted by a stretched exponential. For a given type of grains, the characteristic times of relaxation and of convection are found to be of the same order of magnitude. On the contrary, compaction mechanisms in the media strongly depend on the grain anisotropy. PMID- 15754082 TI - Home-based resistance training improves femoral bone mineral density in women on hormone therapy. AB - This study tested whether moderate resistance training would improve femoral bone mineral density (BMD) in long-term users of hormone therapy with low BMD. The study was a 2-year randomized, controlled, trial (RCT) of moderate resistance training of either the lower extremity or the upper extremity. Eighty-five women participated in a 6-month observation period. The setting was center-based and home-based training. The participants were 189 women aged 59-78 years, with total femur T-scores from -0.8 to -2.8 and on hormone therapy (HT) for a minimum of 2 years (mean 11.8 years); 153 completed the trial. Lower extremity training used weight belts (mean 7.8 kg) in step-ups and chair rises; upper extremity training used elastic bands and dumbbells. Measurements were BMD and body composition [dual-energy X-ray absorptiometry (DXA)], bone turnover markers. Total femoral BMD showed a downward trend during the observation period: 0.35%+/-0.18% (P=0.14). The response to training was similar in the upper and lower groups in the primary outcomes. At 2 years, total femoral BMD increased 1.5% (95% CI 0.8% 2.2%) in the lower group and 1.8% (95% CI 1.1%-2.5%) in the upper group. Trochanter BMD increased 2.4% (95% CI 1.3%-3.5%) in the lower group and 2.5% (95% CI 1.4%-3.6%) in the upper group (for both analyses time effect P<0.001). At 1 year, a bone resorption marker (C-telopeptide) decreased 9% (P=0.04). Bone formation markers, bone-specific alkaline phosphatase, decreased 5% (P<0.001), and N-terminal type I procollagen peptide decreased 7% (P=0.01). Body composition (percent lean and percent body fat) was maintained in both groups. We concluded that long-term moderate resistance training reversed bone loss, decreased bone turnover, increased femur BMD, and maintained body composition. The similarity of response in upper and lower groups supports a systemic response rather than a site-specific response to moderate resistance training. PMID- 15754084 TI - The dynamics of water in nanoporous silica studied by dielectric spectroscopy. AB - Broad-band dielectric spectroscopy is used to investigate the dynamics of hydration water on the surface of the cylindrical pores of a nanostructured silica material (MCM-41, with pore diameter of 3.2 nm) at various hydrations, in the temperature range 250-150 K. We focus our attention on orientational relaxations that shift from 0.5 MHz at 250 K to less than 1 Hz at 150 K. The measurements distinguish the relaxation of the hydroxyl groups at the surface of silica from the orientational dynamics of hydration water which strongly depends on the degree of hydration. Although it is significantly faster than the dynamics of water in ice, the orientational relaxation of "non-freezing" water has an activation energy comparable to that in ice when the hydration layer is complete and approximately two-molecule thick. PMID- 15754085 TI - Time course of myocardial stromal cell-derived factor 1 expression and beneficial effects of intravenously administered bone marrow stem cells in rats with experimental myocardial infarction. AB - OBJECTIVE: The chemokine stromal cell-derived factor-1 (SDF-1) has been implicated in homing of bone marrow cells to sites of injury. We investigated the time course of myocardial SDF-1 expression and effects of intravenously administered bone marrow mesenchymal stem cells (MSC) in rats with myocardial infarction (MI). METHODS: SDF-1 expression was measured by RT-PCR and Western blot in sham operated or infarcted hearts at 1/2, 1, 2, 4, 8 and 16 days post operation. MSCs from donor rats were labeled with BrdU. A total of 5 x 10(6) cells in 2.5 mL of PBS or equal volume PBS alone were injected through the tail vein at above mentioned time points. The number of the labeled MSCs in the infarcted hearts was counted 3 days post injection. Cardiac function and vessel numbers were assessed 28 days post injection. RESULTS: Myocardial SDF-1 expression increased and peaked at the first day and decreased thereafter post MI and remained unchanged in sham operated hearts. The MSCs enrichment and angiogenesis in the host hearts were more abundant in the 1 day transplantation group than in the other groups (P < 0.01). Cardiac function was only improved in rats received intravenous MSCs injection within 4 days post MI and not affected by PBS injection. CONCLUSIONS: Myocardial SDF-1 expression was increased only in the early phase post MI. MSCs intravenous infused at the early phase of MI were recruited to injured heart, enhanced angiogenesis and improved cardiac function. PMID- 15754086 TI - Changes in expression levels of genes involved in fatty acid metabolism: upregulation of all three members of the PPAR family (alpha, gamma, delta) and the newly described adiponectin receptor 2, but not adiponectin receptor 1 during neonatal cardiac development of the rat. AB - During neonatal cardiac development, the heart changes its substrate preference from glucose to fatty acids. The aim of this study was to investigate the changes in mRNA expression levels of genes involved in the control of cardiac fatty acid metabolism in the transition from neonatal to adult life. METHODS: mRNA expression levels for peroxisome proliferator activated receptor (PPAR) alpha, gamma and delta, PPARgamma co-factor 1 alpha and beta (PGC-1 alpha and beta), 9 cis retinoc-acid-activated receptor alpha, beta and gamma (RXR alpha, beta, gamma), 5'-AMP activated protein kinase (AMPK) alpha1 and alpha2, adiponectin receptor 1 and 2 (AR 1 and AR 2) were measured in heart tissue of neonatal 0-day, 7-day and 21- day old rats. RESULTS: mRNA expression of all three members of the PPAR family were upregulated significantly from day 0 to day 21 (alpha +117%, gamma +133%, delta +203%). In addition, m-RNA expression of all RXR isoforms increased from day 0 to day 7 (alpha +125%, beta +69%; gamma +41%). AR 2 exhibited a small but significant increase in mRNA expression (+ 46%). CONCLUSIONS: We were able to demonstrate for the first time that in addition to PPARalpha, also PPARgamma and delta, as well as all RXR isoforms and AR 2 are upregulated in the heart during neonatal development. PMID- 15754087 TI - Sudden cardiac death in dogs with remodeled hearts is associated with larger beat to-beat variability of repolarization. AB - Increased proarrhythmia in dogs with chronic AV block (AVB) has been explained by ventricular remodeling causing a decrease in repolarization reserve. Beat-to-beat variability of repolarization (BVR) has been suggested to reflect repolarization reserve, in which high variability represents diminished reserve and larger propensity for repolarization-dependent ventricular arrhythmia. A subset of chronic AVB dogs (10%) suffers sudden cardiac death (SCD). With the assumption that repolarization defects constitute a potentially lethal proarrhythmic substrate, we hypothesized that BVR in SCD dogs are larger than in matched control chronic AVB dogs. From a population of 200 chronic AVB dogs, initially two groups were chosen retrospectively: 8 dogs that died suddenly (SCD) and 8 control dogs. Control dogs had a longer lifespan after AVB (10 to 18 weeks) than SCD dogs (5 to 10 weeks). All dogs had undergone electrophysiological testing under anesthesia where ECG, left and right ventricular endocardial monophasic action potentials (MAP) were recorded. BVR was assessed from 30 consecutive beats, illustrated by Poincare plots and was the only parameter discriminating between SCD and control group. All other electrophysiological parameters (RR, QT and MAP durations) were comparable for the two groups. Extending the number of animals and groups confirmed a larger BVR in the SCD group (SCD: 5.1 +/- 2.7; n = 11 versus control: 2.5 +/- 0.4 ms; n = 61; P < 0.05) and showed reverse-use dependence of BVR. In comparison, dogs with acute AVB had low variability (1.3 +/ 0.3 ms; n = 9; P < 0.05 versus chronic AVB). Cardiac electrical remodeling after AVB is associated with an increase in beat-to-beat variability of repolarization. Chronic AVB dogs displaying further elevated variability of repolarization are prone to arrhythmia-related SCD. PMID- 15754088 TI - Sorcin interacts with sarcoplasmic reticulum Ca(2+)-ATPase and modulates excitation-contraction coupling in the heart. AB - Sorcin is a 21.6-kDa Ca(2+) binding protein of the penta-EF hand family. Several studies have shown that sorcin modulates multiple proteins involved in excitation contraction (E-C) coupling in the heart, such as the cardiac ryanodine receptor (RyR2), L-type Ca(2+) channel, and Na(+)-Ca(2+) exchanger, while it has also been shown to be phosphorylated by cAMP-dependent protein kinase (PKA). To elucidate the effects of sorcin and its PKA-dependent regulation on E-C coupling in the heart, we identified the PKA-phosphorylation site of sorcin, and found that serine178 was preferentially phosphorylated by PKA and dephosphorylated by protein phosphatase-1. Isoproterenol allowed sorcin to translocate to the sarcoplasmic reticulum (SR). In addition, adenovirus-mediated overexpression of sorcin in adult rat cardiomyocytes significantly increased both the rate of decay of the Ca(2+) transient and the SR Ca(2+) load. An assay of oxalate-facilitated Ca(2+) uptake showed that recombinant sorcin increased Ca(2+) uptake in a dose dependent manner. These data suggest that sorcin activates the Ca(2+)-uptake function in the SR. In UM-X7. 1 cardiomyopathic hamster hearts, the relative amount of sorcin was significantly increased in the SR fraction, whereas it was significantly decreased in whole-heart homogenates. In failing hearts, PKA phosphorylated sorcin was markedly increased, as assessed using a back phosphorylation assay with immunoprecipitated sorcin. Our results suggest that sorcin activates Ca(2+)-ATPase-mediated Ca(2+) uptake and restores SR Ca(2+) content, and may play critical roles in compensatory mechanisms in both Ca(2+) homeostasis and cardiac dysfunction in failing hearts. PMID- 15754089 TI - Myoclonus of probable spinal origin as a potential side effect of tramadol. A case report. PMID- 15754091 TI - Assessment of hepatic reserve for the indication of hepatic resection. AB - The main goal of liver resection in patients with liver tumor is to cure the patients with the lowest operative risk. Based on our experiences, we report herein, indications of liver resection according to the nature of underlying liver parenchyma. In patients with normal underlying liver parenchyma, major resection does not require specific preoperative assessment. Non-tumorous biopsy is indicated in cases of massive steatosis and/or in patients in whom a complex technical procedure is anticipated. Portal vein embolization (PVE) is considered if the future liver remnant (FLR) is < 30%. In patients with chronic liver disease, major hepatectomy is a risky situation, requiring specific preoperative assessment, including non-tumorous biopsy and PVE which could be associated with transarterial chemoembolization. Absence of hypertrophy of the FLR after PVE is a contraindication to a major hepatectomy. Intraoperative procedures in patients "at risk" require specific techniques, including intermittent clamping, anterior approach with hanging maneuver, and fixation on anatomical position of the remnant liver. PMID- 15754092 TI - Assessment of hepatic reserve for the indication of hepatic resection: how I do it. AB - This author has personally carried out in excess of 700 major hepatic resections for tumor, and runs a unit with a current resection rate of 200 per year, yet uses no scientific tests designed to judge hepatic reserve. In our unit, we have an advantage in that we deal with a northern European population, with a low rate of viral hepatitis, although alcoholism is becoming an increasing feature within our practice and we are dealing with more elderly patients that in the past, and more who have undergone neoadjuvant chemotherapy. In these patients, there appear to be greater risks of postoperative sepsis and slower regeneration. Approximately 65% of our current resection practice is hemihepatectomy or more and the majority is trisectionectomy (extended hepatectomy) and bilateral resection work. Preoperative, operative, and postoperative factors affect the occurrence of postoperative hepatic failure and these aspects are considered. Case series studies are presented to illustrate the incidence of significant hepatic failure we have encountered. PMID- 15754090 TI - Discrepancy between clinical and pathological diagnoses of CBD and PSP. AB - Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are often clinically confused with each other. Moreover, the discrepancy between clinical and pathological diagnoses of CBD and PSP are still controversial. We report here two atypical cases of PSP and CBD. A 73-year old woman was admitted with right hand rigidity, limb kinetic apraxia and cortical sensory loss. Brain atrophy, hypoperfusion and hypometabolism predominantly in the left frontoparietal lobes indicated CBD clinically. Pathological studies revealed neuronal loss and spongy change without ballooned neurons (BN) in the cerebral cortex. Modified Gallyas-Braak (G-B) staining revealed neurofibrillary tangles (NFTs) and tufted astrocytes, indicating pathological diagnosis of PSP. A 75-year old man admitted with vertical gaze palsy, neck dystonia, parkinsonism and dementia. Atrophy of the frontal lobes and tegmentum of the midbrain and symmetrical frontal hypoperfusion in SPECT indicated PSP. However, neuronal loss and BN in the frontal lobes and clusters of astrocytic plaques indicated CBD pathologically. The G-B staining was useful for differentiating between CBD and PSP, but our atypical cases bring up a new issue about differential diagnosis of CBD and PSP. PMID- 15754093 TI - Assessment of hepatic reserve prior to hepatic resection. PMID- 15754094 TI - Assessment of hepatic reserve for indication of hepatic resection: decision tree incorporating indocyanine green test. AB - Preoperative assessment of liver function and prediction of postoperative remaining functional liver parenchymal mass and reserve is of paramount importance to minimize surgical risk, especially in patients with hepatocellular carcinoma (HCC), the majority of whom have liver cirrhosis as a complication. We have established a decision tree for deciding the safe limit of hepatectomy based on three variables: whether ascites is present, the serum total bilirubin level, and the indocyanine green retention rate at 15 minutes (ICGR-15), an indicator of sinusoidal capillarization. In patients who show a sign of decompensated cirrhosis as reflected by an elevated bilirubin value or uncontrollable ascites, hepatectomy is not indicated. In patients without ascites and with normal bilirubin level, the ICGR-15 value becomes the main determinant for the resectability and hepatectomy procedure. Incorporation of ICGR-15 into the decision tree enables patients conventionally classified into Child-Turcotte-Pugh class A or score 5-6 to be subdivided into several groups in which various hepatectomy procedures are feasible: enucleation, limited resection, segmentectomy, mono- to bisectoriectomy, and trisectriectomy. During strict application of this decision tree to 1429 consecutive hepatectomies, of which 685 were performed on HCC patients, during the last 10 years, we encountered only a single mortality. PMID- 15754095 TI - Assessment of hepatic reserve for indication of hepatic resection: how I do it. AB - BACKGROUND/PURPOSE: Hepatic resection may result in liver failure in patients with cirrhotic livers. Preoperative evaluation of liver function in cirrhotic patients, to prevent postoperative liver failure, is very important. METHODS: Sixteen patients with hepatocellular carcinoma in cirrhotic livers were enrolled in this study. Liver function was determined quantitatively by monoethylglycinexylidide (MEGX) formation from the metabolism of lidocaine. The whole liver volume and tumor volume were measured by computed tomographic volumetry. The volume of resected liver was recorded by water displacement. The relationship between liver function and remnant liver volume was determined. RESULTS: A relationship between the percentage remnant liver volume and ratio of MEGX formation after hepatectomy was found. The regression equation was: (postoperative MEGX formation/preoperative MEGX formation) x 100% = (0.688 x percentage remnant liver volume + 0.179) x 100% (r (2) = 0.49). A relationship between MEGX formation after hepatectomy and the international ratio (INR) of prothrombin time was also found. The regression equation was INR = 1.99 - 0.01 x MEGX (r (2) = 0.30). CONCLUSIONS: Post-hepatectomy liver function can be estimated for an individual patient by the appropriate regression equations. Prevention of post-hepatectomy liver failure for patients with cirrhotic livers is feasible. PMID- 15754096 TI - Assessment of hepatic reserve for the indication of hepatic resection: how I do it. AB - Despite the careful selection of cirrhotic patients with hepatic neoplasms, liver resection for these patients remains associated with greater risk than in patients without underlying liver disease. The most rational indications for resective surgery in patients with hepatic neoplasms and cirrhosis are nonprogressive cirrhosis and good functional reserve. Therefore, evaluation of hepatic reserve is mandatory for hepatectomy candidates. Because of the complexity of hepatic function, a single, reliable liver function test is not yet available. However, a good multifactorial system that combines several elements (clinical, laboratory, functional, and volumetric evaluation) does provide sufficient data for determining the safe limits of hepatectomy. PMID- 15754097 TI - Assessment of hepatic reserve for indication of hepatic resection: how I do it. AB - The perioperative outcome of hepatic resection has improved remarkably in recent years with improved surgical techniques and perioperative care. As a result, the indications of hepatic resection have been extended to include patients with borderline liver function, especially those with associated cirrhosis. For such patients, proper preoperative assessment of liver function reserve is essential to ensure a low incidence of postoperative liver failure and mortality. In our center, routine preoperative assessment of hepatic function reserve in all patients includes clinical assessment, liver biochemistry, coagulation profile, platelet count, and Child-Pugh classification. The indocyanine green clearance test is routinely performed for patients with chronic liver disease. For patients with cirrhosis undergoing major hepatectomy, computed tomography volumetry and laparoscopy are helpful in evaluating whether the remnant liver volume is adequate. In selected patients with small remnant liver, preoperative portal vein embolization can be employed to induce hypertrophy of the remnant liver even in the presence of chronic hepatitis and mild cirrhosis. Careful assessment of comorbid conditions of patients and meticulous surgical techniques to reduce bleeding and hypoxic injury to the remnant liver are complementary to the selection of patients with adequate liver function reserve, to minimize operative mortality. PMID- 15754098 TI - How I do it: assessment of hepatic functional reserve for indication of hepatic resection. AB - Liver resection of up to 75% of the total liver volume (TLV) has been regarded as safe in normal livers, but this concept was challenged by the results of living donor hepatectomies. In normal livers or livers with resolved jaundice, hepatectomy of 65% of TLV may be safe, except for patients with an indocyanine green retention rate at 15 min (ICG R15) of over 15%, excessive hepatic steatosis, and age of over 70 years. However, the permissible extent of hepatectomy has been much restricted in cirrhotic livers because most post hepatectomy liver failure (PHLF) has occurred in cirrhotic livers. Our routine protocols for the assessment of functional hepatic reserve (FHR) include biochemical liver function tests, ICG R15, Doppler ultrasonography, and triphasic liver computed tomogram (CT) with volumetry. Blood cell count and gastroesophageal endoscopic findings are taken into consideration for cirrhotic livers, as well as age, diabetes, cardiopulmonary function, and general performance. Preoperative portal vein embolization has been used for safe hepatectomy even in cirrhotic livers. We think that any cirrhotic liver showing optimal FHR should have a remnant liver of 40% of TLV to prevent PHLF. ICG R15 and triphasic CT with volumetry have been the most useful methods for assessment of FHR and determination of hepatectomy extent in our institution. PMID- 15754099 TI - A scoring system for the assessment of the risk of mortality after partial hepatectomy in patients with chronic liver dysfunction. AB - BACKGROUND/PURPOSE: The aim of this study is to evaluate a new scoring system, called the chronic liver dysfunction (CLD) score, for prediction of the surgical risk of partial hepatectomy in patients with chronic liver damage. Morbidity and mortality rates after gastroenterological surgery are high in patients with hepatic cirrhosis. Accordingly, it is very important to assess the surgical risk in such patients before surgery. Although the Child classification (or Child-Pugh score) has been a standard system, it did not always accurately predict patients at the risk of mortality after gastroenterological surgery, especially partial hepatectomy. METHODS: In 1985, we established a new system called the CLD score, reviewing the patients undergoing gastroenterological operations at one hospital. In the present study, we prospectively used the CLD score in 256 consecutive patients with chronic liver dysfunction who were treated surgically by partial hepatectomy, and investigated the usefulness of the CLD score concerning mortality. The results were compared with those of the Child-Pugh score (C-P score). RESULTS: After major hepatectomy, all the patients with CLD score exceeding 1.5 died of hepatic failure. After minor hepatectomy, all the patients with CLD score exceeding 2.5 died of hepatic failure. On the other hand, C-P score did not predict the outcome in these patients. CONCLUSIONS: Compared with the C-P score, which was considered the gold standard scoring system for assessing surgical risk for patients with chronic liver dysfunction, our CLD score provides a more reliable assessment of the risk of partial hepatectomy. PMID- 15754100 TI - Laparoscopic hepatectomy with the hook blade of ultrasonic coagulating shears and bipolar cautery with a saline irrigation system. AB - BACKGROUND: The number of patients who have undergone laparoscopic hepatectomy is small, and the operative procedure is not yet well established. METHODS: We performed laparoscopic hepatectomy in eight patients, using the hook blade of ultrasonic coagulating shears, and bipolar cautery with a saline irrigation system, with minilaparotomy. The operative time, blood loss, and postoperative hospital stay of patients with laparoscopic left lateral segmentectomy were compared with these parameters in ten patients who had had a left lateral segmentectomy with laparotomy. RESULTS: The laparoscopic hepatectomies included seven left lateral segmentectomies and one nonanatomical partial resection of the lateral segment. The mean duration of the operation in these eight patients was 181.1 +/- 44.6 min. The mean amount of blood loss was 177.6 +/- 129.1 ml. Postoperative complications consisted of two cases of bleeding. The mean postoperative hospital stay in all eight patients was 9.88 +/- 4.36 days. The mean duration of operation (185.9 +/- 46.0 min) and mean postoperative hospital stay (9.47 +/- 4.61 days) in the seven patients with laparoscopic left lateral segmentectomies were significantly shorter than these parameters (255.7 +/- 59.4 min and 24.6 +/- 8.82 days) in the ten patients who had had left lateral segmentectomies with laparotomy. The mean amount of blood loss (160.0 +/- 128.9 ml) in the laparoscopic series was less than that (318.5 +/- 192.2 days) in the patients who had had laparotomy. CONCLUSIONS: Laparoscopic hepatectomy with the ultrasonic coagulating shears and bipolar cautery with minilaparotomy was safe, and less invasive than the open procedure, for minor hepatectomy procedures such as left lateral segmentectomy. PMID- 15754101 TI - The significance of intraoperative Doppler ultrasonography in evaluating hepatic arterial flow when assessing the indications for the Appleby procedure for pancreatic body cancer. AB - BACKGROUND/PURPOSE: The Appleby procedure has been used in the surgical treatment of advanced gastric cancer. This procedure consists of a combined resection of the whole of the stomach, the body and tail of the pancreas, and the spleen, as well as resection of the celiac artery. This procedure can also be used for operative resection of cancer of the body of the pancreas. The hepatic arterial flow is supplied from the arcade of the supramesenteric artery. In the past, the extent of hepatic arterial flow after the celiac artery had been clamped was evaluated by finger palpation. However, this is not an objective method. METHODS: Here, we describe two patients with pancreatic body cancer, in one of whom the Appleby procedure was followed. However, in the other, this operation could not be performed because the residual blood supply to the liver would have been inadequate. The hepatic arterial flow was assessed using intraoperative Doppler ultrasonography (US) of the intrahepatic artery (arterial flow of segment 3). RESULTS: In case 1 (a 45-year-old man), the hepatic arterial flow prior to clamping of the celiac artery was 68.4 cm/s, and this flow was reduced to 22.1 cm/s after the clamping. The color and consistency of the liver surface remained good. Because there was adequate hepatic arterial flow after the celiac artery was clamped, the Appleby procedure was performed. However, in case 2 (a 65-year old man), the hepatic arterial flow after the celiac artery had been clamped was reduced from 47.9 cm/s to 14.3 cm/s. The liver surface became dark and the liver shrank. In addition, there were various preoperative medical conditions in this patient. We decided not to proceed with the Appleby procedure. CONCLUSIONS: Intraoperative Doppler US measurement of the hepatic arterial flow is a useful technique, particularly in combination with the assessment of the color and tension of the liver, the age of the patient, and the extent of preoperative comorbidity, in determining whether an Appleby procedure is feasible. PMID- 15754102 TI - Features and management of bile leaks after laparoscopic cholecystectomy. AB - BACKGROUND/PURPOSE: Leakage of bile is one of the troublesome complications after laparoscopic cholecystectomy. METHODS: The present study reviewed our experience with this complication, in order to analyze its characteristics and proper management. RESULTS: Postoperative bile leaks occurred in 23 of 1365 patients (1.7%) undergoing laparoscopic cholecystectomy from July 1990 to May 2002, with the policy of routine operative cholangiography and routine drainage of the gallbladder bed. These patients could be divided into four types. In type 1 (17 patients), bile leakage stopped spontaneously within 3 days (subclinical group). In type 2 (3 patients), the leak continued for longer than 3 days but was controlled by an endoscopic nasobiliary drainage (ENBD tube; minor-leakage group). In type 3 (2 patients), bile leakage continued for longer than 3 days and required open repair (major-leakage group). In type 4 (1 patient), bile leakage started several days after surgery (delayed-leakage group). CONCLUSIONS: It is thought that better understanding of these four types of bile leakage should help in the proper management of this complication. PMID- 15754103 TI - Safety and efficacy of laparoscopic distal pancreatectomy for the treatment of pancreatic disease. AB - BACKGROUND/PURPOSE: There are a few reports of laparoscopic distal pancreatectomy in the literature. We describe our experience with laparoscopic distal pancreatectomy and evaluate the safety and efficacy of the procedure in light of other reported findings. METHODS: A retrospective study was performed of all patients who underwent laparoscopic distal pancreatectomy between April 1996 and December 2002 at Oita University Faculty of Medicine. RESULTS: Laparoscopic distal pancreatectomy was attempted in seven patients (three men and four women) with a mean age of 65 years. One patient was converted to open surgery, and two patients required a hand-assistance procedure. There were no complications in any patients. Median operation time for all seven patients was 300 min, and median blood loss was 330 ml. Median postoperative hospital stay was 12 days (range, 7 to 21 days). CONCLUSIONS: Our limited results, together with reported outcomes, suggest that laparoscopic distal pancreatectomy is safe and effective for selected patients. The potential advantages of this procedure include reduced morbidity and reduced hospital stay. PMID- 15754104 TI - Single-branch resection of the pancreas. AB - The treatment of intraductal papillary mucinous tumors (IPMT) of the pancreas is still controversial. In this report we describe a single-branch resection of the pancreas (SBRP), which is a new method for the removal of branch-type IPMT of the head of the pancreas. A multilobular cystic lesion (50 x 40 mm) in the head of the pancreas was incidentally detected in an asymptomatic 40-year-old man who underwent a routine ultrasound examination. The tumor was carefully removed along the border of the cyst and the normal parenchyma, with complete preservation of the main pancreatic duct and the common bile duct. A pancreatic fistula developed during the postoperative period, but was well-controlled by endoscopic naso pancreatic drainage. SBRP is a technically feasible procedure and this operation represents a minimally invasive alternative to any other segmental resection of the pancreas. PMID- 15754106 TI - A minute nonfunctioning islet cell tumor demonstrating malignant features. AB - We report a patient with a minute nonfunctioning islet cell tumor, 8 mm in diameter, which demonstrated malignant features by histology. The patient was a 43-year-old Japanese woman, who had an elevated carbohydrate antigen (CA) 19-9 level, of 59 U/ml (normal range, <37 U/ml) identified on a health check to rule out malignancies. Ultrasonography and computed tomography demonstrated a well defined pancreatic tumor, 8 mm in diameter, in the body of the pancreas. Serum levels of pancreatic hormones were within normal limits, and thus a tentative diagnosis was nonfunctioning islet cell tumor. The size of the tumor remained unchanged for 1 1/2 years, but, at this time, the serum level of CA19-9 was elevated to 253 U/ml. Segmental pancreatectomy was performed because malignancy could not be ruled out. The resected specimen showed an endocrine tumor invading both the pancreatic parenchyma and the perineural spaces outside the tumor. In general, minute nonfunctioning islet cell tumors have been considered to be completely benign, but the present tumor showed clear malignant features. We might have to take surgical resection into consideration even if the size of such an endocrine tumor is minute. PMID- 15754105 TI - Complete obstruction of the lower common bile duct caused by autoimmune pancreatitis: is biliary reconstruction really necessary? AB - Recent observations suggest that an immune response is involved in the development of chronic pancreatitis. We report a case of autoimmune pancreatitis in a patient who showed complete obstruction of the lower common bile duct. A 63 year-old man was admitted to a local hospital, complaining of appetite loss and back pain. The patient had obstructive jaundice, and percutaneous transhepatic gallbladder drainage was performed. Fluorography through the biliary drainage catheter showed complete obstruction of the lower common bile duct. The patient had no history of alcohol consumption and no family history of pancreatic disease. Physical examination revealed an elastic hard mass palpable in the upper abdomen. Abdominal ultrasound and abdominal computed tomography (CT) scans showed enlargement of the pancreas head. While autoimmune pancreatitis was highly likely, due to the patient's high serum immunoglobulin level, the possibility of carcinoma of the pancreas and/or lower common bile duct could not be ruled out. Laparotomy was performed, and wedge biopsy samples from the pancreas head and body revealed severe chronic pancreatitis with infiltration of reactive lymphocytes, a finding which was compatible with autoimmune pancreatitis. Cholecystectomy and biliary reconstruction, using choledochojejunostomy, were performed, because the complete bile duct obstruction was considered to be irreversible, due to severe fibrosis. After the operation, prednisolone (30 mg/day) was given orally for 1 month, and the entire pancreas regressed to a normal size. Complete obstruction of the common bile duct caused by autoimmune pancreatitis has not been reported previously; this phenomenon provides an insight into autoimmune pancreatitis and provokes a controversy regarding whether biliary reconstruction is needed for the treatment of complete biliary obstruction caused by autoimmune pancreatitis. PMID- 15754107 TI - Effect of a portal venous stent for gastrointestinal hemorrhage from jejunal varices caused by portal hypertension after pancreatoduodenectomy. AB - We report herein the case of a 64-year-old man successfully treated by portal venous stent placement for repeated gastrointestinal bleeding associated with jejunal varices. He was admitted to our hospital with melena 8 years after having a pancreatoduodenectomy for carcinoma of the papilla of Vater. From portogram findings showing severe portal vein (PV) stenosis and dilated collaterals through the jejunal vein of the Roux-en-Y loop, jejunal varices resulting from PV stenosis were suspected as the cause of the melena. A metallic stent was placed in the PV following percutaneous transhepatic PV angioplasty. Although the cure of hemorrhagic jejunal varices caused by PV stenosis is difficult in patients who have undergone major abdominal surgery, patency of the stent in this patient has been maintained for 32 months without gastrointestinal hemorrhage. Metallic stent placement is recommended as a useful treatment for PV stenosis that is less invasive than open surgery. PMID- 15754108 TI - Changes to levels of serum neuron-specific enolase in a patient with small cell carcinoma of the pancreas. AB - Small cell carcinoma (SCC) of the pancreas is a rare disease, with an extremely poor prognosis; only 24 cases have been reported in the literature. However, as some patients have been successfully treated with combination chemotherapy, it is important to obtain both a definite diagnosis and a precise evaluation of the effect of the treatment. A 69-year-old woman presented with an abdominal tumor and pain. She had been observed for sensory neuropathy and swelling of the pancreatic head by the referring doctor over the previous 9 months. The patient was diagnosed with SCC of the pancreas after surgery and had two courses of combination chemotherapy (cisplatin and etoposide). Initially, the tumor disappeared completely on computed tomography (CT) scans, but she died of disease recurrence 3 months after completing the chemotherapy. Changes in serum neuron specific enolase (NSE) levels were monitored constantly during the progress of the disease. NSE levels had already increased above the upper limit of normal 8 months before the patient's admission to our hospital, and levels changed concurrently not only with tumor growth but also subsequently with remission and then relapse of the disease after treatment. These results indicate that NSE is a good marker, both as a diagnostic indicator for SCC of the pancreas and as a means of evaluating response to treatment. PMID- 15754109 TI - Pancreatographic findings in idiopathic acute pancreatitis. AB - BACKGROUND/PURPOSE: Despite extensive evaluation based on clinical history, biochemical tests, and noninvasive imaging studies, the cause of acute pancreatitis cannot be determined in 10 to 30% of patients, and a diagnosis of idiopathic acute pancreatitis is made. The purpose of this study was to clarify the pancreatographic findings in patients with idiopathic acute pancreatitis. METHODS: Endoscopic retrograde cholangiopancreatography (ERCP) was performed in 34 patients with idiopathic acute pancreatitis, and the pancreatographic findings were examined. Patency of the accessory pancreatic duct was examined by dye injection endoscopic retrograde pancreatography (ERP) in 16 of the 34 patients. RESULTS: In 11 patients (32%), the following anatomic abnormalities of the pancreatic or biliary system were demonstrated: complete pancreas divisum (n = 5), incomplete pancreas divisum (n = 2), high confluence of pancreaticobiliary ducts (n = 2), choledochocele (n = 1), and giant periampullary diverticulum (n = 1). Pancreatographic findings were normal in 17 patients. Eleven of these patients were examined by dye-injection ERP, and all were found to have nonpatent accessory pancreatic duct. CONCLUSIONS: Anatomic abnormality of the pancreatic or biliary system is one of the major causes of idiopathic acute pancreatitis. Closure of the accessory pancreatic duct may play a role in the development of idiopathic acute pancreatitis in patients with a normal pancreaticobiliary ductal system. PMID- 15754110 TI - Treating the "wise baby". AB - In a previous examination of Ferenczi's concept of the "Wise Baby" (1996), I had noted both its applications and its limitations in the analytic treatment of an unusually intelligent adult. Ferenczi's concepts of "the origin of intellect in trauma" and of "the wise baby" have often left the indistinct impression of being interrelated phenomena. In this paper, I regard as arguable the notion that very high intelligence is pathological when it is "precocious." This return visit to the territory of the "wise baby" extends Ferenczi's ideas about the "origin" and use of the intellect to include a consideration of what may constitute effective "treatment" for those who suffer from giftedness. PMID- 15754111 TI - Reconsideration of "motherhood" in contemporary Japan. AB - In this paper, the author reconsiders "motherhood" in Japan. This reconsideration is based on Japanese psychoanalytic knowledge and a case study of a woman. As a child she was physically abused by her father, and struggled throughout her life with conflicts with her mother. The Japanese have historically idealized the concept of "motherhood" and maintained that it was possible for women to become the ideal mother for their children. The author maintains that "motherhood" is not dependent only on mothers, but is created and shared by fathers, children and all of society. In psychotherapy, the therapist provides a "motherly" energy to the client and shares the "motherhood" fantasy with the client to a certain extent. The therapist assists the client in the gradual process of abandoning the desire to be loved by the ideal mother and accept "motherhood" from other sources. PMID- 15754113 TI - Father and daughter: historical identification, present attachments and the question of passionate involvement. AB - An extended case description is used to illustrate and explore the notion that some children can identify with parental historical experiences as a defense against oedipal passions and mature involvement. Emphasis is placed on the nature and sequelae of a father-daughter relationship, as a function of the father's Holocaust experiences where he passed as a Christian and his post Holocaust experiences with a daughter born to him after the war. The value of adding analytic group therapy to individual analytic work is also discussed in terms of its significance as a potential activator of otherwise latent material that may lay dormant in the dyadic setting. PMID- 15754112 TI - From benign mirror to demon lover: an object relations view of compulsion versus desire. AB - This article addresses the terrifying power of the father as a dynamic force within the psyche of the female. Its particular focus is on how the psychodynamic form of father internalization within the female effects her creative process and her creative work. Two cases vignettes of female writers are used to illustrate the interaction of traumatizing parental behavior and the daughter's incestuous desire on promoting compulsions and inhibitions in creative work. Writing blocks will be seen as a symptom of trauma and psychic conflict that has taken place in the oedipal and post oedipal years, as opposed to the creative compulsions that result from primal stage preoedipal trauma. PMID- 15754115 TI - Scientific meetings of the American Institute for Psychoanalysis. The history of membership/certification in the American Psychoanalytic Association: old demons, new debates. Presenter: Arnold Richards. Discussant: Kenneth Winarick. Date: May 20, 2004. PMID- 15754118 TI - Association of cyclooxygenase-2 and matrix metalloproteinase-2 expression in human breast cancer. AB - Expression of cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) associates with reduced survival in human breast cancer. COX-2 may be directly involved with mammary carcinogenesis, since expression of COX-2 is sufficient for formation of breast tumors in transgenic mice, and COX-2 selective inhibitors can suppress tumorigenesis in rodent models of breast cancer. MMP-2 is an extracellular matrix degrading proteolytic enzyme that bas been linked to invasion and metastasis. A direct link between COX-2 and MMP-2 may exist, since inhibition of COX-2 activity can result in reduction of MMP-2 expression and activity. In this study we analyzed protein expression of COX-2 and MMP-2 in tissue array specimens of 278 invasive breast cancers by immunohistochemistry. Immunopositivity of these two markers was correlated with each other and with various clinicopathological parameters including survival. We found high COX-2 expression in 30% and high MMP-2 expression in 83% of the breast cancer specimens, and there was a positive association between the expression of these two factors (p = 0.003). It was especially evident that whenever COX-2 expression was high, MMP-2 expression was almost invariably high (95%). Furthermore, high expression of either COX-2 or MMP-2 associated with decreased disease specific survival when compared with the COX-2 or MPP-2 low group (p = 0.026 and p = 0.021, respectively). Taken together, our results indicate that expression of COX 2 protein is associated with expression of MMP-2 protein in human breast cancer and that both COX-2 and MMP-2 are markers of poor prognosis in breast cancer. PMID- 15754119 TI - Comparison of methods to diagnose lymphoedema among breast cancer survivors: 6 month follow-up. AB - One of the more problematic and dreaded complications of breast cancer is lymphoedema. Our objective was to determine the prevalence of lymphoedema 6 months following breast cancer treatment and to examine potential risk factors among a population-based sample of women residing in South-East Queensland (n = 176). Women were defined as having lymphoedema if the difference between the sum of arm circumferences (SOAC) of the treated and untreated sides was >5 cm (prevalence = 11.9%) or >10% (prevalence = 0.6%), their multi- frequency bioelectrical impedance (MFBIA) score was > or =3 standard deviations above the reference impedance score (prevalence = 11.4%), or they reported 'yes' when asked if arm swelling had been present in the previous 6 months (prevalence = 27.8%). Of those with lymphoedema defined by MFBIA, only 35% were detected using the SOAC method (difference > 5 cm), while 65% were identified via the self-report method (i.e., respective sensitivities). Specificities for SOAC (difference > 5 cm) and self-report were 88.5% and 76.9%, respectively. When examining associations between presence of lymphoedema and a range of characteristics, findings also varied depending on the method used to assess lymphoedema. Nevertheless, one of the more novel and significant findings was that being treated on the non dominant, compared to dominant, side was associated with an 80% increased risk of having lymphoedema (MFBIA). Our work raises questions about the use of circumferences as the choice of measurement for lymphoedema in both research and clinical settings, and assesses MFBIA as a potential alternative. PMID- 15754120 TI - Racial differences in the familial aggregation of breast cancer and other female cancers. AB - Although breast cancer familial aggregation has been studied in Caucasians, information for African-Americans is scant. We used family cancer history from the Women's Contraceptive and Reproductive Experiences study to assess the aggregation of breast and gynecological cancers in African-American and Caucasian families. Information was available on 41,825 first and second-degree relatives of Caucasian and 28,956 relatives of African-American participants. We used a cohort approach in which the relative's cancer status was the outcome in unconditional logistic regression and adjusted for correlated data using generalized estimating equations. Race-specific models included a family history indicator, the relative's age, and type. Relative risk (RR) estimates for breast cancer were highest for first-degree relatives, and the overall RR for breast cancer among case relatives was 1.96 (95% CI = 1.68-2.30) for Caucasian and 1.78 (95% CI = 1.41-2.25) for African-Americans. The effect of CARE participants' reference age on their relatives' breast cancer risk was greatest among first degree relatives of African-American patients with RRs (95% CI) for ages <45 and > or =45 of 2.97 (1.86-4.74) and 1.48 (1.14-1.92), respectively. Among Caucasians, first-degree relatives of case subjects were at greater risk for ovarian cancer, particularly relatives younger than 45 years (RR (95% CI) = 2.06 (1.02-4.12)), whereas African-American first-degree relatives of case subjects were at increased cervical cancer risk (RR (95% CI) = 2.17 (1.22-3.85). In conclusion, these racially distinct aggregation patterns may reflect different modes of inheritance and/or environmental factors that impact cancer risk. PMID- 15754121 TI - Efficacy of weekly paclitaxel in patients with docetaxel-resistant metastatic breast cancer. AB - BACKGROUND: Partial cross-resistance to paclitaxel and docetaxel has been demonstrated in pre-clinical studies. PATIENTS AND METHODS: We retrospectively evaluated the efficacy of weekly paclitaxel 80 mg/m(2) in 82 patients with docetaxel-resisitant metastatic breast cancer. Docetaxel resistance was classified into primary resistance, defined as progressive disease while receiving docetaxel, and secondary resistance, defined as progression after achievement of a documented clinical response to docetaxel. Secondary resistance was subclassified according to the interval between the final infusion of docetaxel and the start of weekly paclitaxel into: (1) short interval, < or =120 days, and (2) long interval, >120 days. RESULTS: The response rate of the 82 patients was 19.5% (95% confidence interval, 10.8-27.9%). The response rate according to the docetaxel resistance category was: primary resistance (n = 24), 8.3%; secondary resistance (n = 58), 24.1% (short interval [n = 39], 17.9%, and long interval, [n = 19], 36.8%). The differences in response rates among the three categories were statistically significant (p = 0.0247, Cochran-Mantel Haenszel test). The interval between from the final docetaxel infusion and disease progression were predictors for response of weekly paclitaxel. CONCLUSION: Weekly paclitaxel is modestly effective and safe in docetaxel resistant metastatic breast cancer patients. However, weekly paclitaxel should not be recommended for primary resistance patients with docetaxel. PMID- 15754122 TI - Effect of paroxetine hydrochloride (Paxil) on fatigue and depression in breast cancer patients receiving chemotherapy. AB - BACKGROUND: Fatigue can significantly interfere with a cancer patient's ability to fulfill daily responsibilities and enjoy life. It commonly co-exists with depression in patients undergoing chemotherapy, suggesting that administration of an antidepressant that alleviates symptoms of depression could also reduce fatigue. METHODS: We report on a double-blind clinical trial of 94 female breast cancer patients receiving at least four cycles of chemotherapy randomly assigned to receive either 20 mg of the selective serotonin re-uptake inhibitor (SSRI) paroxetine (Paxil, SmithKline Beecham Pharmaceuticals) or an identical-appearing placebo. Patients began their study medication seven days following their first on-study treatment and continued until seven days following their fourth on-study treatment. Seven days after each treatment, participants completed questionnaires measuring fatigue (Multidimensional Assessment of Fatigue, Profile of Mood States Fatigue/Inertia subscale and Fatigue Symptom Checklist) and depression (Profile of Mood States-Depression subscale [POMS-DD] and Center for Epidemiologic Studies Depression [CES-D]). RESULTS: Repeated-measures ANOVAs, after controlling for baseline measures, showed that paroxetine was more effective than placebo in reducing depression during chemotherapy as measured by the CES-D (p = 0.006) and the POMS-DD (p = 0.07) but not in reducing fatigue (all measures, ps > 0.27). CONCLUSIONS: Although depression was significantly reduced in the 44 patients receiving paroxetine compared to the 50 patients receiving placebo, indicating that a biologically active dose was used, no significant differences between groups on any of the measures of fatigued were observed. Results suggest that modulation of serotonin may not be a primary mechanism of fatigue related to cancer treatment. PMID- 15754123 TI - Soy isoflavone supplementation elevates erythrocyte superoxide dismutase, but not plasma ceruloplasmin in postmenopausal breast cancer survivors. AB - Soy isoflavone antioxidant effects may help prevent breast cancer re-occurrence, but isoflavone estrogen-like actions may increase breast cancer risk. These isoflavone actions can be reflected by effects on two copper enzymes activities, superoxide dismutase 1 (SOD 1), which has antioxidant function relevant to breast cancer prevention, and ceruloplasmin, which has its synthesis up-regulated by estrogen, and for which high values are associated with high breast cancer risk. A soy isoflavone-rich concentrate supplement was examined for effects on these two copper enzyme activities in post-menopausal breast cancer survivors (n = 7) in a crossover design with a placebo (24 days on supplement or placebo; 14 day wash out). The soy concentrate, but not the placebo, increased erythrocyte SOD 1 activities, but not ceruloplasmin activities or protein. The effect on superoxide dismutase activities was not likely due to increased copper intake since analysis of the soy extract showed little copper. The effect on superoxide dismutase was not accompanied by a change in urinary contents of 8-deoxyhydroxyguanosine, a DNA oxidant product, though perhaps this would change with a longer intervention. In summary, in regard to two copper enzyme activities, an isoflavone-rich soy concentrate showed an antioxidant effect considered relevant to breast cancer, but not an effect associated with estrogenic activity and increased breast cancer risk. PMID- 15754124 TI - Development of a novel method for measuring in vivo breast epithelial cell proliferation in humans. AB - Cell proliferation plays an important role in all stages of carcinogenesis. Currently, no safe, direct, in vivo method of measuring breast epithelial cell (BEC) proliferation rates in humans exists. Static immunohistochemical markers of cell proliferation, such as Ki-67 and PCNA indices, have technical limitations including high inter-lab variability, inaccuracy in the presence of agents that cause G1/S cell cycle block and inadequate sensitivity in post-menopausal women with low BEC proliferation rates. We describe here a safe, direct method of measuring BEC proliferation rates in vivo in women using heavy water ((2)H(2)O) labeling coupled with mass spectrometric analysis. Proliferation of normal and tumor BEC was measured from breast tissue biopsies in women undergoing mastectomy (n = 11) and normal BEC from healthy volunteers (n = 16). Women took heavy water (50-150 ml per day) for 1-4 weeks. Pre-menopausal women had significantly higher proliferation rates than post-menopausal women (0.7 +/- 0.1 versus 0.2 +/- 0.1 new cells per day, respectively), and tumor BEC had different proliferation rates than normal BEC from the same breast. The method is analytically reproducible and remains sensitive in the range of low proliferation rates. In summary, this novel method of measuring BEC proliferation in vivo holds promise for assessing the effects of anti-proliferative chemopreventive and chemotherapeutic agents. PMID- 15754125 TI - Cancer incidence in multiple sclerosis and effects of immunomodulatory treatments. AB - Multiple sclerosis (MS) has been linked to reduced rates of cancer prior to the era of immunomodulating treatments. We assessed the incidence of cancer in a cohort of 1338 MS patients and evaluated the effect of exposure to immunomodulatory treatment. Cancer incidence in the MS population was compared with the expected age- and gender-matched incidence rates in the Israeli population for the period 1960-2003. Time-dependant Cox model analysis was used to estimate hazard ratios for glatiramer acetate, beta-interferons (1a and 1-b) and intravenous immunoglobulins (IVIg). Among 892 female MS patients, 15 (1.7%) developed breast cancer, and 31 (3.5%) developed cancers of any type. Seventeen of 446 (3.8%) male MS patients developed cancer. The standardized incidence ratios (SIRs) computed until the time of first immunomodulatory treatment were 0.60 (95% CI, 0.38-0.92, p = 0.02) for all female cancer, and 1.11 (95% CI, 0.64 1.91) for all male cancer. Time-dependent covariate analyses for female breast cancer yielded a relative risk for glatiramer acetate of 3.10 (95% CI, 0.86-11.1) and 0.52 (95% CI, 0.07-4.05) for beta-interferons. For IVIg, the analyses were uninformative. Our findings indicate that cancer incidence is significantly lower in female MS patients than in the general population. Female MS patients treated with glatiramer acetate showed an elevated rate of breast cancer and all MS patients treated with beta-interferons showed an elevated risk of non-breast cancers though not statistically significant (p = 0.122 and 0.072, respectively). Further study is needed to assess possible associations between long-term exposure to the novel immunomodulatory treatments in MS and rate of cancer. PMID- 15754126 TI - Diagnosis, treatment and prognosis of internal mammary lymph node recurrence in breast cancer patients. AB - Recurrences in the internal mammary lymph nodes (IMLN) are very rare, despite the fact that these nodes remain untreated in most patients. The aim of this study was to assess the chance for IMLN recurrence in a large patient series and to get insight into diagnostics, treatment and prognosis of this type of recurrence. Follow-up of nearly 6000 breast cancer patients resulted in the tracing of only six patients with IMLN recurrence. IMLN recurrence was defined as breast cancer recurrence in an internal mammary lymph node without a distant metastasis before the recurrence and confirmed by cytology and/or CT-scan. The time interval between diagnosis of the primary tumor and the recurrence varied between 5 months and 8 years and 6 months. One patient showed no symptoms, the other five all had a swelling and one of them also had pain. The size of the parasternal swelling varied from 30 to 90 mm; in one patient the size was unknown. Treatment resulted in a complete remission in four patients. In five of the six patients distant metastases occurred. The time interval between IMLN recurrence and the diagnosis of distant metastasis varied between 0 and 37 months. One patient was still free of distant metastasis in the time of this study. This large population-based study confirms the almost negligible risk of clinically apparent IMLN recurrence. Considering the high percentage of positive lymph nodes in studies evaluating sentinel node biopsy of the internal mammary chain, it becomes clear that just a fraction of these becomes clinically apparent as a recurrence. In almost all patients with IMLN it is a forerunner of metastatic disease. PMID- 15754127 TI - Radiation dose and risk of soft tissue and bone sarcoma after breast cancer treatment. AB - BACKGROUND: To quantify the risk of soft tissue and bone sarcomas after breast cancer according to the doses and technical modalities of irradiation. METHODS: We followed a cohort of 6597 breast-cancer patients for 8.3 years on average. The number of soft tissue and bone sarcomas was compared to the expected number based on the incidence rates in the general French population. We also estimated the risk of sarcoma according to the radiation dose received at site of the sarcoma in a nested case control study of 14 breast-cancer patients who subsequently developed a sarcoma and 98 controls matched for age at diagnosis of breast cancer, period of initial treatment and length of follow-up. RESULTS: In the cohort-study, 12 women who had initially received radiotherapy treatment developed a bone or soft tissue sarcoma during the follow-up period. The expected number of cases during this period was 1.7 (SIR = 7.0, 95% CI: 3.7-11.7) and the mean annual excess incidence during the same period was 21 per 100,000 person years. The 15-year cumulative incidence of sarcoma was 0.28% (95% CI: 0.10 0.45%). In the case-control study, all the 14 cases had received at least 11.8 Gray at the site of the sarcoma, which was always located in the irradiated field or in the upper ipsilateral extremity of the arm. A dose-effect relationship was observed (p < 0.001). The best fit was obtained for a quadratic dose-response relationship, without a negative exponential term for cell killing at high doses. The risk of sarcoma was 30.6 higher for doses of more than 44 Gray than for doses of less than 15 Gray. CONCLUSIONS: High doses of radiation strongly increase the risk of bone and soft tissue sarcoma. PMID- 15754128 TI - PEA3, AP-1, and a unique repetitive sequence all are involved in transcriptional regulation of the breast cancer-associated gene, mammaglobin. AB - The breast cancer-associated gene mammaglobin is a member of the secretoglobin protein family and has demonstrated its utility as a breast cancer marker. However, the transcriptional regulation of mammaglobin has not been well characterized. In this report, we used luciferase reporter assays to identify the 200 bp directly 5' of the transcriptional start site as the minimal promoter region of mammaglobin. Sequence scanning indicated that two PEA3 transcription sites were possibly involved in mammaglobin transcription. By transfecting a PEA3 expression vector into breast cancer cell lines MDA-MB-415 and MCF-7, we determined that exogenous PEA3 was able to drive transcription. Mutational analysis indicated that each PEA3 site was functional. Our reporter system and electrophorectic mobility shift assays (EMSAs) also identified the involvement of a unique repetitive element in mammaglobin transcription. Finally, AP-1 was determined via luciferase assays to be involved in regulating non-PEA3 dependent transcription. Elucidating these cis-acting elements will impact our understanding of transcription of normal breast and breast cancer-associated genes. PMID- 15754129 TI - The "portrait" of hereditary breast cancer. AB - Five to ten per cent of all breast carcinomas are of hereditary origin. Many of them have been associated to mutations in the BRCA1 and BRCA2 susceptibility genes. No "BRCA3" gene has been found to account for the non-BRCA1/BRCA2 breast cancer (BRCAx) families, and BRCAx tumors are increasingly believed to originate from multiple distinct genetic events. Phenotype studies have questioned the existence of specific "portraits" among hereditary breast carcinomas (HBC). They have shown that most BRCA1 tumors have a "basal (epithelial)-like" aspect, while BRCA2 and BRCAx HBC are more heterogeneous. HBC have also been submitted to genetic analyses, notably with the objective of resolving the heterogeneity of BRCAx lesions. The present review aims to summarize recent data on BRCA1, BRCA2, and BRCAx HBC, including hypotheses on the origin of BRCA1 tumors and their paradoxical relations to estrogen-sensitivity. PMID- 15754130 TI - Improvement in intramammary sentinel lymph node removal using a novel prototype hand held probe during breast conservative surgery. AB - Intramammary sentinel lymph node excision during breast conservative surgery was performed, in this case report, using a prototype intraoperative gamma probe. In contrast to the four axillary sentinel lymph nodes that were subnormal, the excised intramammary sentinel lymph node was massively invaded by cancer cells. Therefore this finding had profound implication for the staging of the tumor and for treatment selection. This case report illustrates that an efficient intraoperative gamma probe is useful to locate and remove intramammary sentinel lymph node in breast cancer patients treated with breast conservation. PMID- 15754131 TI - HER2 codon 655 polymorphism and breast cancer: results from kin-cohort and case control analyses. AB - Several published epidemiologic studies show increased breast cancer risk for carriers of the Val-allele at codon 655 of the HER2 gene. We conducted additional analyses using data from three studies, including case-control analyses stratified on age and kin-cohort analyses using relatives of cases and controls. The results provide additional evidence that HER2 codon 655 genotype may predispose to early-onset breast cancer. PMID- 15754132 TI - Cyclin I is expressed in human breast cancer and closely associated with VEGF and KDR expression. AB - In the present study, cyclin I protein expression in 114 invasive human breast cancers was correlated with cell cycle and angiogenesis-related proteins and clinico-pathological data. A strong association was found between cytoplasmic cyclin I staining and VEGF (p = 0.001) as well as the VEGF receptor KDR (p = 0.001), suggesting a link between cyclin I and angiogenesis. PMID- 15754133 TI - Possible role of CXCR4-mediated chemotaxis in breast cancer patients with central nervous system metastases. PMID- 15754137 TI - The nano-hardness and elastic modulus of sound deciduous canine dentin and young premolar dentin--preliminary study. AB - The purpose of this study was to compare the nano-hardness and elastic modulus among deciduous and permanent dentin, buccal and lingual sides, incisal, center and cervical areas, and outer, middle and inner layers. Three premolars and three deciduous canines were bucco lingually (BL) sectioned, and three deciduous canines were mesio-distally (MD) sectioned parallel to the long axis at the center of the tooth. Hardness (H), plastic hardness (PH) and Young's modulus (Y) were measured using a nano-indentation tester. The H, PH and Y values from the deciduous canine dentin were significantly lower than those from the premolar dentin at most sites. For deciduous canine dentin, the H and PH values of the MD sectioned dentin were significantly higher than those of the BL sectioned dentin in many layers of many areas. Generally deciduous canine dentin had H, PH and Y values that decreased from outer toward the inner layers and significant differences were obtained among the layers in many areas. For MD sectioned deciduous canine and BD sectioned premolar dentin, the H, PH and Y values of the cervical area were significantly lower than those of the incisal and center areas in many layers. It is possible that optimum bonding may require different treatments for deciduous and permanent dentin and perhaps also for different intratooth locations. PMID- 15754138 TI - Preparation and characterization of an electrodeposited calcium phosphate coating associated with a calcium alginate matrix. AB - A new way of optimizing osteoconduction of biomaterials is to bring to them biological properties. In this work, we associated a novel release system with an electrodeposited calcium phosphate (CaP) coated titanium alloy Ti6Al4V. The characterization of this material was performed by means of light microscopy, scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM) and X-ray energy dispersive spectroscopy (EDXS). The electrodeposited CaP coating was a tricalcium phosphate, and the release system was composed of microcapsules entrapped in an alginate film. We observed that the alginate matrix had a close contact with the coating. An intermediate layer containing calcium and phosphorus appeared at the interface between the alginate matrix and the CaP coating. These results allowed us to conclude that the association of two techniques, i.e. electrodeposition followed by deposition of a calcium alginate matrix, led to the elaboration of a new biomaterial. PMID- 15754139 TI - Residual monomers released from glass-fibre-reinforced composite photopolymerised in contact with bone and blood. AB - PURPOSE: The aim of this study was to determine the quantity of residual monomers of glass fibre-reinforced composite released into water from the composite that had been photopolymerized in contact with bone and blood. MATERIALS AND METHODS: E-glass fibre reinforced composite (FRC) made of E-glass fibre veil and the bis GMA-TEGDMA-PMMA resin system was used in the study. In the first group, pieces of non-polymerised FRC were photopolymerised (40 s) in air which influenced the oxygen inhibited resin layer (positive control). In the second group, the FRC was polymerized between two glass plates allowing both surfaces to be well polymerized (negative control). In the test groups, the FRC was polymerized in contact with bone or in contact with blood. FRC specimens from all four groups were incubated in three milliliters of deionised water at 37 degrees C for three days. At the end of the incubation period, the residual monomers were extracted from the water with dichloromethane, and the residual monomers of TEGDMA and bis GMA quantitatively analysed by HPLC. The degree of monomer conversion was measured by FTIR from the surface of the test specimen. Differences between the groups were analysed using one-way ANOVA (p < 0.05). RESULTS: The total quantity of residual monomers released from FRC polymerized in contact with bone was lower (0.55 wt%) than in the positive control group (0.97 wt%) (p = 0.021), and only slightly exceeded that of the negative control group (0.42 wt%) (p = 0.717). The total quantity of monomers released from FRC polymerized in contact with blood was at the level of the negative control group. The main residual monomer released was TEGDMA. The surfaces of the positive and negative controls showed a clear difference between the degree of monomer conversion, 34.0 and 62.8%, respectively, when analysed with FTIR (p < 0.001). CONCLUSION: The surface of the bone or contact with blood did not significantly inhibit the photoinitiated free radical polymerisation of the dimethacrylate monomer system of the FRC. PMID- 15754140 TI - Effect of three surface conditioning methods to improve bond strength of particulate filler resin composites. AB - The use of resin-based composite materials in operative dentistry is increasing, including applications in stress-bearing areas. However, composite restorations, in common with all restorations, suffer from deterioration and degradation in clinical service. Durable repair alternatives by layering a new composite onto such failed composite restorations, will eliminate unnecessary loss of tooth tissue and repeated insults to the pulp. The objective of this study was to evaluate the effect of three surface conditioning methods on the repair bond strength of a particulate filler resin-composite (PFC) to 5 PFC substrates. The specimens were randomly assigned to one of the following surface conditioning methods: (1) Hydrofluoric (HF) acid gel (9.5%) etching, (2) Air-borne particle abrasion (50 microm Al2O3), (3) Silica coating (30 microm SiOx, CoJet-Sand). After each conditioning method, a silane coupling agent was applied. Adhesive resin was then applied in a thin layer and light polymerized. The low-viscosity diacrylate resin composite was bonded to the conditioned substrates in polyethylene molds. All specimens were tested in dry and thermocycled (6.000, 5 55 degrees C, 30 s) conditions. One-way ANOVA showed significant influence of the surface conditioning methods (p < 0.001), and the PFC types (p < 0.0001) on the shear bond strength values. Significant differences were observed in bond strength values between the acid etched specimens (5.7-14.3 MPa) and those treated with either air-borne particle abrasion (13.0-22.5 MPa) or silica coating (25.5-41.8 MPa) in dry conditions (ANOVA, p < 0.001). After thermocycling, the silica coating process resulted in the highest bond values in all material groups (17.2-30.3 MPa). PMID- 15754141 TI - MBCP biphasic calcium phosphate granules and tissucol fibrin sealant in rabbit femoral defects: the effect of fibrin on bone ingrowth. AB - An ageing population implies an increase in bone and dental diseases, which are in turn a source of numerous handicaps. These pathologies are an expensive burden for the European health system. As no specific bioactive materials are efficient enough to cope with this burden, we have to develop an injectable, mouldable, self-hardening bone substitute to support bone tissue reconstruction and augmentation. New, highly bioactive and suitable biomaterials have been developed to replace bone grafts in orthopedic revision and maxillofacial surgery for bone augmentation. These mouldable, self-hardening materials are based on the association of MBCP Biphasic Calcium Phosphate Granules and Tissucol Fibrin Sealant. The in vivo evaluation of ingrowth in relation to the composite was made in an experiment on rabbits. The results indicate that in the presence of fibrin sealant, newly-formed bone developed at a small distance from the surface of the calcium phosphate ceramic. Two different bone apposition processes were identified. Without the fibrin component (MBCP group), bone rested directly on the surface of the granules. This observation is commonly described as osteoconduction in calcium phosphate materials. On the contrary, the presence of the fibrinogen component seemed to modify this standard osteoconduction phenomenon: the newly-formed bone essentially grew at a distance from the surface of the granules, on the fibrillar network, and could be considered as an inductive phenomenon for osteogenic cell differentiation from mesenchymal stem cells. PMID- 15754142 TI - Microneedles coated with porous calcium phosphate ceramics: effective vehicles for transdermal delivery of solid trehalose. AB - Trehalose (alpha-D-glucopyranosyl-alpha-D-glucopyranoside) is recognized as a promising fast-dissolving solid reservoir capable of stabilizing the native structure of proteins and suitable for loading with a wide variety of bioactive substances. Currently, there is a growing interest in developing cost-effective methods for immobilizing solid trehalose on arrays of microneedles for delivering protein-based and DNA-based vaccine to the epidermis. In the present work, micro porous calcium phosphate coatings were used to provide a biocompatible interface with a large surface area for the effective immobilization of trehalose on microneedles. Calcium phosphate coatings with varying degrees of porosity were electrochemically synthesized on the tips of stainless steel acupuncture needles and loaded with solid trehalose. Skin experiments were designed to determine the ability of micro-porous calcium phosphate coatings to deliver solid trehalose into epidermis without breaking during insertion. The mechanical performance of the coatings was assessed by inserting the tips of the coated needles into human skin to an average depth of 100-300 microm and then removing them for analysis by scanning electron microscopy. Microporous calcium phosphate coatings loaded with trehalose effectively breached the stratum corneum and allowed direct access to the epidermis without breaking and without stimulating nerves in deeper tissues. PMID- 15754143 TI - Influence of light-curing procedures and photo-initiator/co-initiator composition on the degree of conversion of light-curing resins. AB - OBJECTIVE: The hypothesis that the degree and rate of conversion can be modified favourably by using different light-curing procedures and different photo initiator/co-initiator combinations was tested. METHOD: A photo-initiator (0.02 mM/g resin); either camphorquinone (CQ) or 1-phenyl-1,2-propanedione (PPD), was mixed with bisGMA:TEGDMA (50:50 by weight). In addition, a co-initiator (0.04 mM/g resin); either N,N-dimethyl-p-aminobenzoic acid ethylester (DABE), N,N cyanoethylmethylaniline (CEMA), or 2-dimethylaminoethyl methacrylate (DMAEMA), was added. These six combinations were subjected to three curing conditions (standard curing, soft-start curing or LED curing). The conversion levels (DC) were determined with differential scanning calorimetry (DSC). The DSC results were analysed using a general linear model (GLM) and Duncan's multiple range test and regular t-test. RESULTS: The fastest conversion initially was obtained by standard curing, followed by LED curing and soft-start curing. After 40 s of curing, conventional curing and soft-start curing produced a higher DC than LED curing. However, strong interactions occurred between the different variables (curing method, initiator and co-initiator). Initially, CQ was more efficient than PPD, but after 40 s, this difference was insignificant. CONCLUSION: By using soft-start curing and an appropriate photo initiator/co-initiator combination it is possible to achieve slow curing and a high DC at within a curing time of 40 s. PMID- 15754144 TI - Preparation and bactericidal property of MgO nanoparticles on gamma-Al2O3. AB - After being impregnated in the solution containing Mg(NO3)2 x 6H2O, gamma-Al2O3 was dried and calcined at 500 degrees C which results in the production of a bactericide, highly dispersed MgO loaded on the surface of gamma-Al2O3. The threshold value of the monolayer dispersion of MgO on gamma-Al2O3 is 14.97%, and MgO crystal formed when the load amount beyond this value. The samples with different load amount were characterized by X-ray Diffraction (XRD), Low temperature N2 adsorption-desorption, Inductively Coupled Plasma (ICP) and High Resolution Scan Electronic Microscope (SEM). The results showed that these MgO microcrystals are highly dispersed and have regular size in the range of 4 to 10.8 nm. The specific surface, pore volume and pore size of the sample decreases with the increase of load amount. It is demonstrated that gamma-Al2O3 with highly dispersed MgO on the surface is efficient bactericide, and the one with 20% load amount of MgO can kill more than 99% bacteria and spore cells. PMID- 15754145 TI - Search for ideal biomaterials to cultivate human osteoblast-like cells for reconstructive surgery. AB - In this study we cultured human osteoblast-like cells on 16 different biomaterials to find an optimal biomaterial for subsequent use in reconstructive surgery. The tested biomaterials can be divided into five groups: collagen-based membranes of bovine, equine or calf origin, tricalcium phosphate based membranes (alpha and beta), hyaluronic acid based, anorganic bovine bone and anorganic silicone-based membranes. Cell proliferation and cell colonization (Environmental Scanning Electron Microscope, ESEM) analysis were performed. The results of the study demonstrated that four of the examined biomaterial/cell constructs showed a very good proliferation rate and cell density: No. 3 (Tissue Vlies), No. 7 (Sepra film), No. 16 (Biobrane) and No. 17 (Biomend). No favourable group of biomaterials was noticeable. Moreover, the results indicate that these four biomaterials as a part of bone constructs are the best tools for engineering new bone tissue. In contrast, biomaterials No. 19a (Bio-Oss) and 19b (Bio-Oss Collagen) showed the lowest proliferation rates. The result of No. 19b was improved by treatment in the perfusion chamber for 48 h as well as by additional use of vacuum. The present study is an important base for further analysis of biomaterials and consequently for the development of tissue engineering. PMID- 15754146 TI - Nano-scaled hydroxyapatite/polymer composite III. Coating of sintered hydroxyapatite particles on poly(4-methacryloyloxyethyl trimellitate anhydride) grafted silk fibroin fibers. AB - A novel composite coupling between nano-scaled hydroxyapatite (HAp) particles and poly[4-methacryloyloxyethyl trimellitate anhydride (4-META)]-grafted silk fibroin (SF) through ionic interaction was synthesized. The weight gain of poly(4-META) by graft-polymerization increased with increasing the reaction time, eventually reaching a plateau value of about 20 wt%. The HAp nano-particles were adsorbed equally and dispersively on the treated SF fiber surface. The HAp content in the composite was 4.554 wt% +/- 0.098 (n = 4), confirmed by thermogravimetry (TG). This synthetic system requires no heat to connect HAp to SF and is useful when applying to non-heat-resistant polymers. The L-929 cell-adhesion test shows that the HAp/SF composite improves bioactivity compared to the original SF. PMID- 15754147 TI - Comparative study of apatite formation on CaSiO3 ceramics in simulated body fluids with different carbonate concentrations. AB - Apatite formation on CaSiO3 ceramics was investigated using two different simulated body fluids (SBF) proposed by Kokubo (1990) and Tas (2000) and three sample/SBF (S/S) ratios (1.0, 2.5 and 8.3 mg/ml) at 36.5 degrees C for 1-25 days. The CaSiO3 ceramic was prepared by firing coprecipitated gel with Ca/Si = 0.91 at 1400 degrees C. The bulk density was 2.14 g/cm3 and the relative density about 76%. The two SBF solutions contain different concentrations of HCO3- and Cl- ions, the concentrations of which are closer to human blood plasma in the Tas SBF formulation than in the Kokubo formulation. The pH values in the former solution are also more realistic. The CaSiO3 ceramics show apatite formation in SBF (Kokubo) after soaking for only 1 day at all S/S ratios whereas different phases were formed at each S/S ratio in SBF (Tas). The crystalline phases formed were mainly apatite at S/S = 1.0 mg/ml, carbonate-type apatite at 2.5 mg/ml and calcite at 8.3 mg/ml. At higher S/S ratios the increase in the Ca concentration became higher while the P concentration became lower in the reacted SBF. These changes in SBF concentrations and increasing pH occurred at higher S/S ratios, producing more favorable conditions in the SBF for the formation of carbonate bearing phases, finally leading to the formation of calcite instead of apatite in the higher HCO3- ion concentration SBF (Tas). Apatite is, however, formed in the lower HCO3- ion concentration SBF (Kokubo) even though the Ca and P concentrations change in a similar manner to SBF (Tas). PMID- 15754149 TI - [Angina pectoris and the normal coronary angiogram]. PMID- 15754148 TI - Surface modification of starch based blends using potassium permanganate-nitric acid system and its effect on the adhesion and proliferation of osteoblast-like cells. AB - The surface modification of three starch based polymeric biomaterials, using a KMnO4/HNO3 oxidizing system, and the effect of that modification on the osteoblastic cell adhesion has been investigated. The rationale of this work is as follows--starch based polymers have been proposed for use as tissue engineering scaffolds in several publications. It is known that in biodegradable systems it is quite difficult to have both cell adhesion and proliferation. Starch based polymers have shown to perform better than poly-lactic acid based materials but there is still room for improvement. This particular work is aimed at enhancing cell adhesion and proliferation on the surface of several starch based polymer blends that are being proposed as tissue engineering scaffolds. The surface of the polymeric biomaterials was chemically modified using a KMnO4/HNO3 system. This treatment resulted in more hydrophilic surfaces, which was confirmed by contact angle measurements. The effect of the treatment on the bioactivity of the surface modified biomaterials was also studied. The bioactivity tests, performed in simulated body fluid after biomimetic coating, showed that a dense film of calcium phosphate was formed after 30 days. Finally, human osteoblast like cells were cultured on unmodified (control) and modified materials in order to observe the effect of the presence of higher numbers of polar groups on the adhesion and proliferation of those cells. Two of the modified polymers presented changes in the adhesion behavior and a significant increase in the proliferation rate kinetics when compared to the unmodified controls. PMID- 15754150 TI - Normal coronary arteries: clinical implications and further classification. AB - The term "chest pain with normal coronary arteries" encompasses a large number of different cardiac pathophysiological abnormalities, including impairment of coronary flow reserve, endothelial dysfunction, and early atherosclerosis that, in most cases, cannot be readily differentiated one from the other. To study early coronary atherosclerosis, physicians must look beyond contrast filled arteries (so called lumenology). Angiograms cannot evaluate the vessel wall, plaque distribution and composition or other morphology. Plaques are often angiographically not visible due to their small size and compensatory enlargement (outward remodeling) of the coronary arteries. As a result, the search for an underlying atherosclerotic process remains ongoing. Available clinical studies showed that many patients with chest pain and normal angiography have early atherosclerosis as documented by intravascular ultrasound imaging, reduced coronary flow reserve and coronary endothelial dysfunction. Additional studies showed that patients presenting with normal coronary angiography have recurrent coronary events at long-term follow up. Research to determine if improved diagnosis and treatment of quantitatively low degrees of atherosclerosis lead to improved outcomes of patients with normal angiography should be undertaken. PMID- 15754151 TI - Normal coronary physiology assessed by intracoronary Doppler ultrasound. AB - There is a natural need to measure flow in normal as well as diseased coronary arteries since the reestablishment of normal flow is the aim of most coronary interventions. Miniaturization has enabled positioning both proximal and distal within the coronary anatomy and improved accuracy of Doppler velocity measurements. Measurement of coronary flow reserve adds functional data to coronary angiographic anatomic description, and quantitation of flow, achieved by combining Doppler velocity. Although new techniques involving transthoracic Doppler and magnetic resonance imaging are currently being developed and refined to measure coronary flow velocity and volume flow noninvasively, the role of intracoronary Doppler remains secure particularly for the coronary interventional physician. PMID- 15754152 TI - Angina pectoris with a normal coronary angiogram. AB - Angina in the setting of a normal angiogram (NOCAD) occurs in 20-30% of patients undergoing coronary angiography. The etiologies of NOCAD can be anatomically classified into three groups: epicardial disease, coronary microvascular dysfunction, and noncoronary disease. Epicardial disease resulting in NOCAD includes endothelial dysfunction, coronary artery spasm, and coronary artery bridging. Microvascular dysfunction may be secondary to hypertension, cardiomyopathy, infiltrative disease, valvular disease, or idiopathic. Noncoronary artery disease states involving other organs systems such as the pulmonary, gastrointestinal, or musculoskeletal systems can also result in NOCAD. This review focuses on the coronary etiologies of NOCAD. The pathophysiology of disease is discussed as well as a systematic diagnostic strategy. Potential therapeutic options and prognosis are also reviewed. PMID- 15754153 TI - [Hypertensive microvascular disease]. AB - Arterial hypertension is a major cardiovascular risk factor and leads to vascular as well as to myocardial manifestations. Particularly hypertensive microvascular disease is of great importance. Major clinical manifestations of hypertensive heart disease are symptoms of coronary insufficiency with typical angina pectoris, but also heart failure (systolic or diastolic dysfunction) and arrhythmia. Different non-invasive and invasive procedures are available for screening. For ultimate quantitative assessment of the coronary reserve, invasive procedures are still required. Beside lowering blood pressure primary therapeutic target is to reverse cardiac manifestations of arterial hypertension using specific therapeutic algorithms. PMID- 15754154 TI - [Normal coronary angiography with myocardial bridging: a variant possibly relevant for ischemia]. AB - Approximately 20-30% of patients with cardiac chest pain have a normal coronary angiogram. In about 5% of these patients, a myocardial bridge can be identified. The characteristic feature is systolic compression of an epicardial vessel, usually the left anterior descending artery (LAD), with the angiographic "milking effect". Using modern imaging techniques, such as intravascular ultrasound (IVUS), intracoronary Doppler ultrasound (ICD) and intracoronary pressure wires, the pathophysiological consequence of myocardial bridging could be established. While previously considered a clinically insignificant variant, ICD recordings demonstrated an increased flow velocity in the tunneled segment. Frame-by-frame IVUS analysis revealed a delayed relaxation after systolic compression, which may extend significantly into diastole. This explains both the impaired coronary flow reserve and ischemia. In IVUS, a circular or eccentric rhythmic compression of the vessel is visible, which may be partial or complete. Latest computed tomography technology can also be used to visualize myocardial bridging noninvasively. Provocation tests, such as application of nitroglycerin, orciprenaline, dobutamine or atrial stimulation, may augment systolic compression and explain symptoms and the beneficial effect of beta-blockers. In severe cases (i.e. limiting symptoms with ischemia despite medication), surgical myotomy may be performed after careful appraisal of the benefit-risk ratio. A high restenosis and complication rate associated with coronary stenting precludes a general recommendation of this interventional approach. Whether drug-eluting stents help to overcome this limitation remains to be shown. Long-term prognosis is good, although previous investigations have been performed in a limited number of cases only. PMID- 15754155 TI - Role of endothelial dysfunction and insulin resistance in angina pectoris and normal coronary angiogram. AB - Angina pectoris and a normal coronary angiogram or cardiac syndrome X is a heterogeneous syndrome that probably encompasses different pathophysiological entities. Patients affected by cardiac syndrome X are often women presenting with severe, invalidating chest pain. However, there is a significant discrepancy among the severity of symptoms, the lack of hemodynamic evidence of myocardial ischemia and the relatively benign long-term prognosis. The vascular endothelium has numerous important functions, including the regulation of vascular tone, blood flow and permeability, secreting both vasorelaxing and vasoconstricting factors. It has been found that both endothelium and non-endothelium-mediated coronary blood flow are impaired in patients with cardiac syndrome X. Interestingly, it has been shown that impaired nitric oxide-dependent vasodilation could increase coronary microvessel tone and produce spasm. It has also been reported that circulating endothelin-1 levels are elevated with a direct relationship between endothelin-1 levels and impaired coronary flow reserve in these patients. In addition, patients with high endothelin-1 levels showed a time onset of chest pain during exercise significantly lower compared to patients with low endothelin-1 concentrations. Moreover, the nitric oxide/endothelin-1 ratio was found decreased in patients with cardiac syndrome X and endothelin-1 levels were also positively correlated with fasting asymmetric dimethylarginine levels. All in all, these data suggest a role of endothelial dysfunction as a cause of regional myocardial and peripheral blood flow abnormalities. Further studies are necessary to characterize the prevailing mechanisms determining alterations in nitric oxide/endothelin-1 pathway in these patients, in order to find new therapies able to improve both quality of life and prognosis. PMID- 15754156 TI - Psychosocial factors associated with noncardiac chest pain and cardiac syndrome X. AB - Cardiac syndrome X, the triad of angina pectoris, positive exercise electrocardiogram (ECG) for myocardial ischemia and angiographically smooth coronary arteries, is associated with increased psychological morbidity, debilitating symptomatology and a poor quality of life. Patients with noncardiac chest pain (NCCP) are often similarly affected. The psychological morbidity noted among this patient population has been linked with a number of psychosocial factors, including impaired social support, traumatic life events, the negative impact of menopause among female sufferers, and an awareness of a family history of coronary heart disease (CHD). Cognitive behavioral therapy (CBT), group support, physical activity and relaxation techniques have been investigated as treatments for psychological morbidity among this patient group with varying degrees of success. While clinicians should be aware of the psychological aspect of patients with NCCP and cardiac syndrome X, further research is needed in order to establish a comprehensive physiological and psychological treatment regimen. PMID- 15754157 TI - Prognostic value of noninvasive stressing modalities in patients with chest pain and normal coronary angiogram. AB - Risk stratification of patients with recurrent chest pain and normal coronary angiogram is a relevant but still definitely unsolved clinical problem. In this article the relative value of mostly used noninvasive stress testing modalities is reviewed. In addition, future perspectives derived from alternative pathophysiological insights and new diagnostic approaches are briefly discussed. PMID- 15754158 TI - Historical aortic Starr-Edwards prosthesis - a pitfall for interventional treatment of complex CAD? PMID- 15754159 TI - [Analysis of systolic function of the left ventricle]. PMID- 15754161 TI - Accuracy of biopsy needle navigation using the Medarpa system--computed tomography reality superimposed on the site of intervention. AB - The aim of this work was to determine the accuracy of a new navigational system, Medarpa, with a transparent display superimposing computed tomography (CT) reality on the site of intervention. Medarpa uses an optical and an electromagnetic tracking system which allows tracking of instruments, the radiologist and the transparent display. The display superimposes a CT view of a phantom chest on a phantom chest model, in real time. In group A, needle positioning was performed using the Medarpa system. Three targets (diameter 1.5 mm) located inside the phantom were punctured. In group B, the same targets were used to perform standard CT-guided puncturing using the single-slice technique. The same needles were used in both groups (15 G, 15 cm). A total of 42 punctures were performed in each group. Post puncture, CT scans were made to verify needle tip positions. The mean deviation from the needle tip to the targets was 6.65+/ 1.61 mm for group A (range 3.54-9.51 mm) and 7.05+/-1.33 mm for group B (range 4.10-9.45 mm). No significant difference was found between group A and group B for any target (p>0.05). No significant difference was found between the targets of the same group (p>0.05). The accuracy in needle puncturing using the augmented reality system, Medarpa, matches the accuracy achieved by CT-guided puncturing technique. PMID- 15754160 TI - Oral contrast agents for small bowel distension in MRI: influence of the osmolarity for small bowel distention. AB - To assess the effect of the osmolarity for small bowel distension in MRI, ten volunteers ingested at two separate occasions negative oral contrast agents with different quantity and osmolarity: (1) a water solution combined with 2.0% sorbitol and 0.2% locus bean gum (LBG) with a quantity of 1500 ml and an osmolarity of 148 mOsmol/l, (2) a water solution combined with 2.0% sorbitol and 2.0% barium sulphate with a quantity of 1000 ml and an osmolarity of 194 mOsmol/l. Small bowel distension was quantified on coronal 2D-TrueFISP images by measuring the small bowel diameters. There were no statistically significant differences in mean small bowel diameter between both contrast agents. The mean small bowel distension was 19.2 mm after ingestion of 1500 ml of sorbitol-LBG solution and 19.0 mm after ingestion of 1000-ml sorbitol-barium sulphate solution. Furthermore, all volunteers found the ingestion of 1000-ml solution more pleasant than the 1500-ml solution. The ingestion of 1000 ml of sorbitol barium sulphate solution led to a sufficient small bowel distension compared to 1500 ml of sorbitol-LBG solution. The side effect rate of both solutions was low. Based on these data, we recommend a quantity of 1000 ml of sorbitol-barium sulphate solution as an alternative for 1500-ml sorbitol-LBG solution for optimal bowel distension. PMID- 15754162 TI - Radiologic-pathologic correlations of intratumoral tissue components in the most common solid and cystic renal tumors. Pictorial review. AB - This paper describes the correlation between US-CT images and pathologic findings in the most common solid and cystic renal tumors, to better differentiate malignant and benign renal masses. Several intratumoral tissue components present correlation with US-CT images. Macroscopic components, corresponding to necrotic, hemorrhagic and cystic changes, are identified by visual analysis of the gross specimen, while microscopic components are identified by histopathologic analysis. Microscopic components are classified as cellular [(1) high cellularity with poor extracellular stroma, +/-high nucleus-cytoplasm ratio+/-high uniformity in tumoral cells dimensions; (2) low cellularity with large extracellular stroma+/-low nucleus-cytoplasm ratio+/-low uniformity in tumoral cells dimensions], stromal [(1) fibrotic; (2) fibrovascular; (3) fibromyxoid], vascular related to neoangiogenesis, necrotic [(1) coagulative; (2) colliquative; (3) hemorrhagic], calcific, and adipose. PMID- 15754163 TI - The relationship between the MRI features of mild osteoarthritis in the patellofemoral and tibiofemoral compartments of the knee. AB - The aim of this work was to demonstrate the relationship between osteoarthritic changes seen on magnetic resonance (MR) images of the patellofemoral (PF) or tibiofemoral (TF) compartments in patients with mild osteoarthritis (OA) of the knee. MR images of the knee were obtained in 105 sib pairs (210 patients) who had been diagnosed with OA at multiple joints. Entry criteria included that the degree of OA in the knee examined should be between a Kellgren and Lawrence score of 2 or 3. MR images were analyzed for the presence of cartilaginous lesions, bone marrow edema (BME) and meniscal tears. The relationship between findings in the medial and lateral aspects of the PF and TF compartments was examined. The number of cartilaginous defects on either side of the PF compartment correlated positively with number of cartilaginous defects in the ipsilateral TF compartment (odds ratio, OR, 55, confidence interval, CI, 7.8-382). The number of cartilaginous defects in the PF compartment correlated positively with ipsilateral meniscal tears (OR 3.7, CI 1.0-14) and ipsilateral PF BME (OR 17, CI 3.8-72). Cartilaginous defects in the TF compartment correlated positively with ipsilateral meniscal tears (OR 9.8, CI 2.5-38) and ipsilateral TF BME (OR 120, CI 6.5-2,221). Osteoarthritic defects lateralize or medialize in the PF and TF compartments of the knee in patients with mild OA. PMID- 15754164 TI - Myocardial viability assessment in patients with highly impaired left ventricular function: comparison of delayed enhancement, dobutamine stress MRI, end-diastolic wall thickness, and TI201-SPECT with functional recovery after revascularization. AB - This study compared different magnetic resonance imaging (MRI) methods with Tl(201) single photon emission computerized tomography (SPECT) and the "gold standard" for viability assessment, functional recovery after coronary artery bypass grafting (CABG). Twenty patients (64+/-7.3 years) with severely impaired left ventricular function (ejection fraction [EF] 28.6+/-8.7%) underwent MRI and SPECT before and 6 months after CABG. Wall-motion abnormalities were assessed by stress cine MRI using low-dose dobutamine. A segment with a nonreversible defect in Tl(201)-SPECT and a delayed enhancement (DE) in an area >50% of the entire segment, as well as an end-diastolic wall thickness <6 mm, was defined as nonviable. The mean postoperative EF (n=20) improved slightly from 28.6+/-8.7% to 32.2+/-12.4% (not significant). Using the Tl(201)-SPECT as the reference method, end-diastolic wall thickness, MRI-DE, and stress MRI showed high sensitivity of 94%, 93%, and 84%, respectively, but low specificities. Using the recovery of contractile function 6 months after CABG as the gold standard, MRI-DE showed an even higher sensitivity of 99%, end-diastolic wall thickness 96%, stress MRI 88%, and Tl(201)-SPECT 86%. MRI-DE showed advantages compared with the widely used Tl(201)-SPECT and all other MRI methods for predicting myocardial recovery after CABG. PMID- 15754166 TI - [Therapy for systemic urothelial cancer recurrence]. AB - Detailed data on treating systemic recurrences of urothelial cancer after radical cystectomy and/or radical upper tract surgery are sparse. This review describes the data available from two large clinical phase III trials, one on systemic chemotherapy for metastatic/inoperable urothelial cancer and the other on adjuvant systemic chemotherapy after radical cystectomy for locally advanced bladder cancer. In addition, an overview of contemporary series on metastasectomy for urothelial cancer with palliative as well as curative intent is presented. The impact of therapeutic options for the treatment of systemic urothelial cancer recurrences such as radiotherapy, surgery or systemic first and second line therapy represented by conventional chemotherapy and contemporary target therapy are discussed. PMID- 15754165 TI - Radiofrequency thermal ablation of liver tumors. AB - Radiofrequency ablation (RFA) of liver tumors was first proposed in 1990. New technologies enable us to produce liver thermal lesions of approximately 3-3.5 cm in diameter; RFA has consequently become an emerging percutaneous therapeutic option both for small hepatocellular carcinoma (HCC) and for non-resectable liver metastases, mainly from colorectal cancer. New devices (for example, triplet of cooled needles, wet needles) and combined therapies (tumor ischemia and RFA) have made it possible to treat large tumors. RFA can be carried out by a percutaneous, laparoscopic or laparotomic approach. Percutaneous RFA can be performed with local anaesthesia and mild sedation; deep sedation or general anaesthesia are also used. The guidance system is generally represented by ultrasound. CT or MR examinations are the more sensitive tests for assessing therapeutic results. The series of patients treated with RFA allow the technique to be considered as effective and safe, achieving a relatively high rate of cure in properly selected cases; it should be classified as curative/effective treatment for HCC, replacing percutaneous ethanol injection. The complication rate of RFA is low but not negligible; key elements in a strategy to minimize them are identified. PMID- 15754167 TI - [Therapy for overactive detrusor using propiverine]. AB - BACKGROUND: Our aim was to evaluate the efficacy and tolerability of propiverine hydrochloride (propiverine) in daily practice and to check the risk-benefit relation using previously collected data on 4,390 patients. PATIENTS AND METHODS: A total of 2,932 patients with symptoms of overactive bladder were treated with propiverine over a period of 12 weeks using a multicentre post marketing surveillance. At three visits (inclusion, after 4 weeks, after 12 weeks), parameters from the micturition diary (incontinence episodes, frequency of micturition, micturition volume) were recorded. RESULTS: The number of incontinence episodes during daytime decreased during therapy from 3.6+/-3.8 to 1.2+/-2.3. The number of episodes at night decreased from 1.5+/-2.1 to 0.4+/-0.8 (both P<0.001). The mean volume per micturition improved during therapy (from 142.7 ml to 213.3 ml; +49.5%; P<0.0001). Some 85% of the investigators judged the efficacy of propiverine to be good or very good, 2.1% as not sufficient. The most frequent adverse event was dry mouth (17.3% of the patients after 12 weeks) mostly with low severity. More than 70% of the patients reported good or very good tolerability. Only 0.6% of the patients reported insufficient tolerability. PMID- 15754168 TI - Correlation between symptoms and radiological findings in patients with chronic rhinosinusitis: an evaluation study using the Sinonasal Assessment Questionnaire and Lund-Mackay grading system. AB - Computerised tomography (CT) scans are routinely performed prior to endoscopic sinus surgery (ESS) in order to confirm the diagnosis, assess the response to medical treatment and provide important landmarks for surgery. However, the correlation between the findings of CT scans and the patient's symptoms remains ambiguous. The objective was to assess the correlation between preoperative symptom scores using the validated Sino-Nasal Assessment Questionnaire and CT scores in patients undergoing ESS. Twenty-two patients who attended otolaryngology clinics with chronic rhinosinusitis (CRS) were prospectively enrolled in the study. All of these patients completed Sinonasal Questionnaires (SNAQ) before they had ESS. Their CT scans were scored blindly by the surgeon and a radiologist following Lund-Mackay grading. Patients with high preoperative SNAQ scores had high or low CT scores. The same applied for patients with low preoperative SNAQ scores. There was no statistically significant correlation between SNAQ and Lund-Mackay scores (P = 0.5). However, there was a very strong correlation (P < 0.001) between the scoring of scans by the surgeon and the radiologist, while there was a moderate degree of discrepancy in the grading of anterior ethmoid sinuses and osteo-meatal complexes. Morbidity of patients with CRS cannot be predicted from the magnitude of changes in their CT scans. Significant inter-rater variability exists in the grading of pathological changes in anterior ethmoid sinus and osteo-meatal complex as recorded in CT scans. PMID- 15754170 TI - Extending the clinicopathological spectrum of neurofilament inclusion disease. AB - We describe features of a patient that broadens the clinical and pathological spectrum of neurofilament inclusion disease (NFID). The patient was a 52-year-old man with a 5--6 year history of progressive, asymmetrical spastic weakness of the upper and lower extremities; L-DOPA-unresponsive parkinsonism; and SPECT evidence of asymmetrical frontoparietal and basal ganglia hypoperfusion. The brain had marked frontoparietal parasagittal cortical atrophy, including the motor cortex, with histopathological evidence of neurofilament- and alpha-internexin immunoreactive neuronal inclusions. The corticospinal tract had degeneration, but there was minimal lower motor neuron pathology. There was also severe neuronal loss and gliosis in the posterolateral putamen and the substantia nigra, mimicking multiple system atrophy; however, glial cytoplasmic inclusions were not detected with alpha-synuclein immunohistochemistry. This case extends the clinical and pathological spectrum of NFID to include cases with predominant parkinsonian and pyramidal features. PMID- 15754169 TI - Cytokeratin expression pattern in congenital and acquired pediatric cholesteatoma. AB - Pediatric cholesteatoma can be classified as congenital or acquired based on clinical criteria. We studied the expression patterns of five distinctive cytokeratins in both types of cholesteatoma in order to improve understanding of their pathogenesis and origin. A comparable expression pattern for CK10, CK14, CK18, CK19 and 34betaE12 antigens was found in the matrix of congenital and acquired pediatric cholesteatoma. Our results demonstrate that congenital and acquired pediatric cholesteatoma exhibit an identical cytokeratin distribution pattern, suggesting that they share a common origin. Therefore, it seems possible that a portion of the so-called "acquired" cholesteatoma may actually originate from advanced congenital cholesteatoma with secondary destruction of the tympanic membrane in the pediatric patient population. PMID- 15754171 TI - Postural stabilization from fingertip contact II. Relationships between age, tactile sensibility and magnitude of contact forces. AB - In the present report, we extend our previous observations on the effect of age on postural stabilization from fingertip contact (Exp Brain Res 157 (2004) 275) to examine the possible influence of sensory thresholds measured at the fingertip on the magnitude of contact forces. Participants (young, n=25, 19-32 years; old, n=35, 60-86 years) underwent psychophysical testing of the right index finger to determine thresholds for spatial acuity, pressure sensitivity and kinesthetic acuity. Spatial acuity was determined from the ability to detect gaps of different widths, while Semmes-Weinstein monofilaments were used for pressure sensitivity. Kinesthetic acuity was determined by asking participants to discriminate plates of different thicknesses using a thumb-index precision grip. These tests were selected on the basis that each reflected different sensory coding mechanisms (resolution of spatial stimuli, detection of mechanical forces and integration of multi-sensory inputs for hand conformation) and thus provided specific information about the integrity and function of mechanoreceptive afferents innervating the hand. After log transformation, thresholds were first examined to determine the influence of age (young and old) and gender (male, female) on tactile acuity. Sensory thresholds were then entered into multiple linear regression models to examine their ability to predict fingertip contact forces (normal and tangential) applied to a smooth surface when subjects stood with eyes closed on either a firm or a compliant support surface. As expected, age exerted a significant effect (p<0.01) on all three thresholds, but its impact was greater on spatial acuity than on pressure sensitivity or kinesthetic acuity. Gender had a marginal impact on pressure sensitivity thresholds only. The regression analyses revealed that tactile thresholds determined at the index fingertip accounted for a substantial proportion of the variance (up to 30%) seen in the contact forces deployed on the touch-plate, especially those exerted in the normal direction. The same analyses further revealed that much of the variance explained by the models arose from inter-individual differences in tactile spatial acuity and not from differences in pressure sensitivity or in kinesthetic acuity. Thus, of all three tests, the spatial acuity task was the most sensitive at detecting differences in hand sensibility both within and between age groups and, accordingly, was also better at predicting the magnitude of fingertip forces deployed for postural stabilization. Since spatial acuity is critically dependent upon innervation density, we conclude that the degree of functional innervation at the fingertip was likely an important factor in determining the capacity of older participants to use contact cues for stability purposes, forcing the most affected individuals to exert unusually high pressures in order to achieve stabilization in the presence of reduced tactile inputs arising from contact with the touched surface. PMID- 15754172 TI - Physical practice induces excitability changes in human hand motor area during motor imagery. AB - The present study was undertaken to investigate the effects of physical practice on excitability changes in human primary motor cortex (M1) during motor imagery (MI). Using different intensities of transcranial magnetic stimulation (TMS), we examined changes in the motor evoked potential (MEP) of the first dorsal interosseous (FDI) muscle with and without MI, and before and after physical practice. On comparing results for MEPs recorded before and after physical practice, the difference between the MEP amplitudes observed at rest and during MI only increased at higher TMS intensities. This finding indicates a physical practice-dependent increase of the higher threshold recruitment of corticospinal tract neurons (CTNs), consistent with synchronization for efficient movement, and provides evidence that neural mechanisms of MI depend not only on the type of movement but also on the extent of the motor adaptation (the physical practice). These present findings also show the benefit of MI and highlight beneficial neural mechanisms related to the activation of M1 during MI. In other words, MI may reflect functional changes of M1 that are similar to the changes observed after physical practice. PMID- 15754173 TI - The effect of rest breaks on human sensorimotor adaptation. AB - We have studied the effect of rest breaks on sensorimotor adaptation to rotated visual feedback in a pointing task. Adaptive improvement was significantly poorer after 1-s breaks than after 5-40-s breaks, with no significant difference among the latter break durations. The benefit of >1-s breaks emerged soon after the onset of adaptation, and then remained steady throughout the adaptation, retention (next day), and persistence (no feedback) phases. This pattern of findings indicates that break-induced facilitation is not a result of strategic adjustments, motivation, or recovery from fatigue, but rather to consolidation of previously acquired sensorimotor recalibration rules. PMID- 15754174 TI - Persistence of inter-joint coupling during single-joint elbow flexions after shoulder fixation. AB - Single-joint elbow flexions are associated with muscle activity at the shoulder that opposes interaction torques arising from rotation of the elbow. We have previously shown that this activity is linearly related to elbow muscle torque and is robust in the presence of novel dynamic loads. Here we examined this relationship in the context of shoulder joint fixation. We tested the hypothesis that after mechanically fixing the shoulder the relationship between shoulder muscle activity and elbow muscle torque will be preserved. In contrast, proposals in which energetic variables are optimized predict that shoulder muscle activity should cease. Subjects performed single-joint elbow flexions in a horizontal plane while interacting with the KINARM robotic exoskeleton. After repeated movements with the shoulder joint fixed we observed a slight and gradual decrease in the activity of pectoralis major relative to movements in which the shoulder was free to rotate. However the strength of the coupling between the shoulder and elbow did not change after shoulder fixation. This is consistent with our previous findings and suggests that the nervous system maintains this inter-joint coupling relationship even when activity at the fixed joint is no longer needed for movement accuracy. PMID- 15754175 TI - Cholinergic cortical circuits in Parkinson's disease and in progressive supranuclear palsy: a transcranial magnetic stimulation study. AB - To investigate the involvement of the cortical cholinergic system in patients with Parkinson's disease (PD) and with progressive supranuclear palsy (PSP), we performed a comparative study of the short latency afferent inhibition (SAI) in ten patients with PD, in eight patients with PSP and in 15 healthy subjects. Six of the PD patients and four of the PSP patients had dementia. SAI was significantly increased in the PD patients, whereas it was not significantly different between PSP patients and the normal controls. Our findings demonstrate that the excitability of the motor cortex is differentially modulated by sensory afferents in PD and PSP and may indicate that the mechanisms of cholinergic dysfunction are different between the two diseases. PMID- 15754176 TI - Mirror, mirror on the wall: viewing a mirror reflection of unilateral hand movements facilitates ipsilateral M1 excitability. AB - Primary motor cortex (M1) excitability is modulated by both ipsilateral limb movement and passive observation of movement of the contralateral limb. An interaction of these effects within M1 may account for recent research suggesting improved functional recovery of the impaired arm following stroke by viewing a mirror reflection of movements of the unimpaired arm superimposed over the (unseen) impaired arm. This hypothesis was tested in the present study using single-pulse transcranial magnetic stimulation (TMS) in eight neurologically healthy subjects. Excitability of M1 ipsilateral to a phasic, unilateral hand movement was measured while subjects performed paced (1 Hz), unilateral index finger-thumb opposition movements. Motor evoked potentials (MEPs) were obtained from the inactive first dorsal interosseous (FDI) in each of four viewing conditions: Active (viewing the active hand), Central (viewing a mark positioned between hands), Inactive (viewing the inactive hand) and Mirror (viewing a mirror reflection of the active hand in a mirror oriented in the mid-sagittal plane) and with both hands at rest (Rest). MEPs were significantly enhanced during ipsilateral hand movement compared with the Rest condition (P<0.05). Largest MEPs were obtained in the Mirror condition, and this was significant compared with both the Inactive and Central viewing conditions (P<0.05). There was no difference between the dominant and non-dominant hand. Excitability of M1 ipsilateral to a unilateral hand movement is facilitated by viewing a mirror reflection of the moving hand. This finding provides neurophysiological evidence supporting the application of mirror therapy in stroke rehabilitation. PMID- 15754177 TI - Relative reward processing in primate striatum. AB - Rewards are often not only valued according to their physical characteristics but also relative to other available rewards. The striatum (caudate nucleus, putamen, ventral striatum including nucleus accumbens) is involved in the organization of movement and the processing of reward information. We studied the activity of single striatal neurons in macaques that were presented with different combinations of two rewards. We found in nearly half of the investigated neurons that the processing for one reward shifted, relative to the other rewards that were available in a given trial block. The relative reward processing concerned all forms of striatal activity related to reward-predicting visual stimuli, arm movements and reception of rewards. The observed changes may provide a neural basis for the known shifts in valuation of rewarding outcomes relative to known references. PMID- 15754178 TI - Concurrent adaptations of left and right arms to opposite visual distortions. AB - Previous research has shown that subjects can adapt with either arm to an opposite visual distortion, and the two adaptive states can then be used in sequence to control the respective arm. To extend this finding, we exposed the left and right arms of our subjects to opposite-directed rotations of the visual field alternately for 20 s each, and determined the time-course of adaptation, as well as aftereffects without visual feedback under uni- and bimanual conditions. Our data confirm that two adaptive states can co-exist in the sensorimotor system, one for each arm. We further found that the time-course of adaptive improvement was similar for both arms, that the improvement was present as early as the first movement after a change of arm and discordance, and that the magnitude of adaptation was similar to control data yielded by a single arm and discordance. Taken together, these findings suggest that the two adaptive states were formed concurrently, and without mutual interference. We also observed significant aftereffects. They were smaller but still appreciable under bimanual conditions; the two arms moved at the same time in different directions even though they were aimed at a common visual target. This outcome indicates that the two adaptive states were not merely of a strategic nature, but rather changed the rules by which sensory information was transformed into motor outputs; it also suggests that the two states not only co-exist, but can also be engaged concurrently in movement control. The reduced aftereffects observed under bimanual conditions can be attributed to the well-known phenomenon of bimanual coupling, which is unrelated to adaptation. PMID- 15754179 TI - Nicotine suppresses the P13 auditory evoked potential by acting on the pedunculopontine nucleus in the rat. AB - We identified a potential novel site of action for nicotine (NIC) since (a) systemic injection of NIC led to a dose-dependent decrease in the amplitude of the sleep state-dependent, vertex-recorded, P13 midlatency auditory evoked potential (generated by the reticular activating system, RAS), (b) localized injections of a nicotinic receptor antagonist into the pedunculopontine nucleus (PPN, the cholinergic arm of the RAS) blocked the effects of systemic NIC on the P13 potential (a measure of level of arousal), and (c) localized injection of a nicotinic receptor agonist into the PPN also led to a decrease in the amplitude of the P13 potential, an effect blocked by PPN injection of a nicotinic receptor antagonist. There were minor changes in the manifestation of the startle response (SR) at the concentrations used; however, NIC did decrease the hippocampal N40 potential, although its effects were not affected by antagonist or agonist injections into the PPN. These results suggest a potential mechanism underlying the anxiolytic effects of NIC-suppression of the cholinergic arm of the RAS. PMID- 15754181 TI - Impairment of skilled forelimb use after ablation of striatal interneurons expressing substance P receptors in rats: an analysis using a pasta matrix reaching task. AB - Local injection of substance P (SP)-saporin can cause selective ablation of striatal interneurons expressing SP receptors (SPR). In this study, we evaluated quantitatively the impairment of skilled forelimb use after unilateral ablation of the striatal interneurons using a pasta matrix reaching task in rats. We found a significant decrease of the number of the pasta pieces (uncooked spaghetti) retrieved using the paw of the experimental side contralateral to the ablation, whereas the number of the pasta pieces retrieved using the paw of the intact side increased significantly. These findings, with our previous reports, suggest that the modulation of the cortico-striato-entopeduncular direct pathway by striatal interneurons is important for maintaining normal basal-ganglia control for skilled forelimb movements. PMID- 15754180 TI - Saccadic instabilities and voluntary saccadic behaviour. AB - Primary gaze fixation is never perfectly stable but can be interrupted by involuntary, conjugate saccadic intrusions (SI). SI have a high prevalence in the normal population and are characterised by a horizontal fast eye movement away from the desired eye position, followed, after a variable duration, by a return saccade or drift. Amplitudes are usually below 1 degrees and they often exhibit a directional bias. The aim of the present study was to investigate the aetiology of SI in relation to saccadic behaviour. It was hypothesised that if SI resulted from deficits in the saccadic system (i.e. reduced inhibitory mechanisms), changes in voluntary saccade behaviour may be apparent and related to SI frequency. To examine this, synchrony (no gap), gap, overlap and antisaccade tasks were conducted on ten normal subjects. No significant correlations were found between SI frequency and voluntary saccade latencies, the percentage of express saccades, or the percentage of antisaccade errors. In addition, no significant correlations were found between SI directional biases and saccade latency directional biases, express saccade biases or antisaccade error biases. These results suggest that an underlying alteration to saccadic behaviour is unlikely to be involved in SI production, and that the SI command signal may arise from the influence of attention on an intact saccadic system. Specifically, descending corticofugal signals relating to attention level and orientation may alter the balance between fixation and saccade generation, so determining SI characteristics. PMID- 15754182 TI - Humans use continuous visual feedback from the hand to control both the direction and distance of pointing movements. AB - Vision of the hand during reaching provides dynamic feedback that can be used to control movement. We investigated the relative contributions of feedback about the direction and distance of the hand relative to a target. Subjects made pointing movements in a 3-D virtual environment, in which a small sphere provided dynamic visual feedback about the position of their unseen fingertip. On a subset of trials, the position of the virtual fingertip was smoothly shifted by 2 cm during movement, either (1) in the direction of movement, which would require adjustments to the distance moved, or (2) orthogonal to the direction of movement, which would require adjustments to the direction moved. Despite not noticing the perturbations, subjects adjusted their movements to compensate for both types of visual shifts. Corrective responses to direction perturbations were observed within 117 ms, and response latencies were invariant to movement speed and perturbation onset time. Initial corrections to distance perturbations were smaller and appeared after longer delays of 130-200 ms, and both the speed and magnitude of responses were reduced for early onset perturbations. Simulations of a feedback control model that optimally integrates visual information over time show that the results can be explained by differences in the sensory noise levels in the visual dimensions relevant for direction and distance control. PMID- 15754183 TI - [Treatment of chronic pain in the internist's practice]. AB - Chronic pain is one of the most frequent complaints in outclinic supply. Careful anamnesis and physical examination are in the focus of diagnostic procedure. Their results will give strong evidence for the underlying pathogenesis. Nevertheless, the perception of pain remains completely subjective and can be only measured by asking the patient. Therefore, a multitude of validated instruments has been developed ranging from the simple intensity rating scales up to sophisticated questionnaires. Both, the diagnostic and therapeutic procedures should adhere to the bio-psycho-social concept of pain. Medical treatment should be executed in accordance to the WHO Guidelines for Cancer Pain Treatment. But for the majority of patients, multidisciplinary concepts are required including psychotherapy and iatrophysics. PMID- 15754184 TI - Trichloroethylene degradation by butane-oxidizing bacteria causes a spectrum of toxic effects. AB - The physiological consequences of trichloroethylene (TCE) transformation by three butane oxidizers were examined. Pseudomonas butanovora, Mycobacterium vaccae, and Nocardioides sp. CF8 utilize distinctly different butane monooxygenases (BMOs) to initiate degradation of the recalcitrant TCE molecule. Although the primary toxic event resulting from TCE cometabolism by these three strains was loss of BMO activity, species differences were observed. P. butanovora and Nocardioides sp. CF8 maintained only 4% residual BMO activity following exposure to 165 microM TCE for 90 min and 180 min, respectively. In contrast, M. vaccae maintained 34% residual activity even after exposure to 165 microM TCE for 300 min. Culture viability was reduced 83% in P. butanovora, but was unaffected in the other two species. Transformation of 530 nmol of TCE by P. butanovora (1.0 mg total protein) did not affect the viability of BMO-deficient P. butanovora cells, whereas transformation of 482 nmol of TCE by toluene-grown Burkholderia cepacia G4 caused 87% of BMO-deficient P. butanovora cells to lose viability. Together, these results contrast with those previously reported for other bacteria carrying out TCE cometabolism and demonstrate the range of cellular toxicities associated with TCE cometabolism. PMID- 15754185 TI - [Annular erythematous plaques]. PMID- 15754186 TI - [Critical incident reporting systems in (CIRSs) anesthesia. Flaw or culture]. PMID- 15754188 TI - [Exposure of hospital personnel to sevoflurane]. AB - BACKGROUND: Occupational exposure to volatile anaesthetics cannot be completely avoided even in modern operating theatres. In 1997, the staff exposure during balanced anaesthesia in our hospital was low (sevoflurane 0.49 ppm; N(2)O 11.5 ppm). In 1999, N(2)O was completely omitted at our hospital, therefore, exposure to volatile anaesthetics, namely sevoflurane, might have increased. METHODS: Environmental exposure was measured by photoacoustic infrared spectrometry. To determine the personal exposure and to compare it with environmental exposure, 14 anaesthetists wore diffusion samplers near their breathing zones for 1 week. RESULTS: The median environmental concentration of sevoflurane was between 0.09 and 0.21 ppm in central operating theatres and between 0 and 24.8 ppm in intervention rooms. The median personal concentration was 0.19 ppm. CONCLUSION: The occupational exposure to volatile anaesthetics is not higher using sevoflurane alone compared to the combination of sevoflurane and N(2)O. In addition, the data acquired from environmental and personal measurements showed similar results. PMID- 15754187 TI - [Errors and risks in perioperative thrombolysis therapy]. AB - Until recently, perioperative thrombolysis has always been thought to be a contraindication, because of the risk of bleeding complications. However, many case reports now show that thrombolytic drugs can be successfully used in the perioperative period. Possible indications for perioperative thrombolysis are massive pulmonary embolism and cardiopulmonary resuscitation which are also important causes of cardiac arrest. Thrombolysis can be the decisive therapeutical option, e.g. when normal cardiopulmonary resuscitation measures are not successful. Nevertheless, even against the background of many positive case reports the indication for perioperative thrombolysis should be a case-specific decision as the frequency of complications depends on the surgical intervention and must be weighed against the possible benefits of early thrombolysis. For perioperative thrombolysis there are no data available showing a beneficial effect of one particular therapeutic regimen. In the literature it is suggested that thrombolysis should be performed as quickly as possible, because the risk of bleeding complications depends more on the duration of thrombolysis than on the dosage or the thrombolytic drug itself. PMID- 15754189 TI - Cloning and characterization of an HDZip I gene GmHZ1 from soybean. AB - By using cDNA-AFLP, we analyzed a recombinant inbred line population of soybean that was derived from a soybean mosaic virus (SMV) resistant cultivar Kefeng No.1 and a susceptible cultivar Nannong 1138-2. One hundred and eight fragments showing polymorphism between SMV resistant and susceptible pools were identified. One fragment w27 was 96 bp in length and showed homology to homeobox ggth with a coding region of 738 bp, encoding a protein of 245 amino acids. The genomic sequence analysis defined an intron of 521 bp in the coding region. GmHZ1 was characterized by the presence of a homeodomain (HD) with a closely linked leucine zipper motif (Zip). Southern blot analysis indicated that there was a single copy of GmHZ1 in the soybean genome. When inoculated with SMV strain N3, resistant and susceptible varieties showed reduced and increased expression of the GmHZ1, respectively. The fusion protein of GmHZ1 with GFP was targeted only in nucleus. Yeast two hybrid studies revealed that the GmHZ1 had transcriptional activation activity and can form homodimer. GmHZ1 can bind two 9-bp pseudopalindromic elements (CAAT(A/T)ATTG and CAAT(C/G)ATTG) with different affinity. Using GUS as a reporter gene, GmHZ1 was proved to be a transcriptional activator and enhanced GUS expression by binding with the two elements in plant cells. These results indicate that the GmHZ1 may have a transcriptional activator function in plant response to SMV infection. PMID- 15754190 TI - Nitric oxide in plants: the biosynthesis and cell signalling properties of a fascinating molecule. PMID- 15754191 TI - Visual pigment spectra of the comma butterfly, Polygonia c-album, derived from in vivo epi-illumination microspectrophotometry. AB - The visual pigments in the compound eye of the comma butterfly, Polygonia c album, were investigated in a specially designed epi-illumination microspectrophotometer. Absorption changes due to photochemical conversions of the visual pigments, or due to light-independent visual pigment decay and regeneration, were studied by measuring the eye shine, i.e., the light reflected from the tapetum located in each ommatidium proximal to the visual pigment bearing rhabdom. The obtained absorbance difference spectra demonstrated the dominant presence of a green visual pigment. The rhodopsin and its metarhodopsin have absorption peak wavelengths at 532 nm and 492 nm, respectively. The metarhodopsin is removed from the rhabdom with a time constant of 15 min and the rhodopsin is regenerated with a time constant of 59 min (room temperature). A UV rhodopsin with metarhodopsin absorbing maximally at 467 nm was revealed, and evidence for a blue rhodopsin was obtained indirectly. PMID- 15754192 TI - Unusual complication of rectopexy with polypropylene mesh. PMID- 15754193 TI - Travelling wave patterns in a model of the spinal pattern generator using spiking neurons. AB - The aim of this study is to produce travelling waves in a planar net of artificial spiking neurons. Provided that the parameters of the waves--frequency, wavelength and orientation--can be sufficiently controlled, such a network can serve as a model of the spinal pattern generator for swimming and terrestrial quadruped locomotion. A previous implementation using non-spiking, sigmoid neurons lacked the physiological plausibility that can only be attained using more realistic spiking neurons. Simulations were conducted using three types of spiking neuronal models. First, leaky integrate-and-fire neurons were used. Second, we introduced a phenomenological bursting neuron. And third, a canonical model neuron was implemented which could reproduce the full dynamics of the Hodgkin-Huxley neuron. The conditions necessary to produce appropriate travelling waves corresponded largely to the known anatomy and physiology of the spinal cord. Especially important features for the generation of travelling waves were the topology of the local connections--so-called off-centre connectivity--the availability of dynamic synapses and, to some extent, the availability of bursting cell types. The latter were necessary to produce stable waves at the low frequencies observed in quadruped locomotion. In general, the phenomenon of travelling waves was very robust and largely independent of the network parameters and emulated cell types. PMID- 15754194 TI - Immunonutrition in the critically ill: from old approaches to new paradigms. PMID- 15754195 TI - Alveolar fibrinolytic capacity suppressed by injurious mechanical ventilation. AB - OBJECTIVE: To investigate the effect of mechanical ventilation on alveolar fibrinolytic capacity. DESIGN AND SETTING: Randomized controlled animal study in 66 Sprague-Dawley rats. SUBJECTS AND INTERVENTIONS: Test animals received intratracheal fibrinogen and thrombin instillations; six were killed immediately (fibrin controls), and the others were allocated to three ventilation groups (ventilation period: 225 min) differing in positive inspiratory pressure and positive end-expiratory pressure, respectively: group 1, 16 cmH2O and 5 cmH2O (n=17); group 2, 26 cmH2O and 5 cmH2O (n=16); group 3, 35 cmH2O and of 5 cmH2O (n=17). Ten animals that had not been ventilated served as healthy controls. MEASUREMENTS AND RESULTS: After animals were killed, we measured D-dimers, plasminogen activator inhibitor (PAI) 1, and tumor necrosis factor alpha in the bronchoalveolar lavage fluid and calculated lung weight and pressure/volume (P/V) plots. The median D-dimer concentration (mg/l) decreased with increasing pressure amplitude (192 in group 1, IQR 119; 66 in group 2, IQR 107; 29 in group 3, IQR 30) while median PAI-1 (U/ml) increased (undetectable in group 1; 0.55 in group 2, IQR 4.55; 3.05 in group 3, IQR 4.85). PAI-1 level was correlated with increased lung weight per bodyweight (Spearman's rank correlation 0.708). Tumor necrosis factor alpha concentration was not correlated with PAI-1 level. CONCLUSIONS: Alveolar fibrinolytic capacity is suppressed during mechanical ventilation with high pressure amplitudes due to local production of PAI-1. PMID- 15754196 TI - Pulse pressure variations to predict fluid responsiveness: influence of tidal volume. AB - OBJECTIVE: To evaluate the influence of tidal volume on the capacity of pulse pressure variation (DeltaPP) to predict fluid responsiveness. DESIGN: Prospective interventional study. SETTING: A 31-bed university hospital medico-surgical ICU. PATIENTS AND PARTICIPANTS: Sixty mechanically ventilated critically ill patients requiring fluid challenge, separated according to their tidal volume. INTERVENTION: Fluid challenge with either 1,000 ml crystalloids or 500 ml colloids. MEASUREMENTS AND RESULTS: Complete hemodynamic measurements including DeltaPP were obtained before and after fluid challenge. Tidal volume was lower than 7 ml/kg in 26 patients, between 7-8 ml/kg in 9 patients, and greater than 8 ml/kg in 27 patients. ROC curve analysis was used to evaluate the predictive value of DeltaPP at different tidal volume thresholds, and 8 ml/kg best identified different behaviors. Overall, the cardiac index increased from 2.66 (2.00-3.47) to 3.04 (2.44-3.96) l/min m(2) ( P <0.001). It increased by more than 15% in 33 patients (fluid responders). Pulmonary artery occluded pressure was lower and DeltaPP higher in responders than in non-responders, but fluid responsiveness was better predicted with DeltaPP (ROC curve area 0.76+/-0.06) than with pulmonary artery occluded pressure (0.71+/-0.07) and right atrial (0.56+/-0.08) pressures. Despite similar response to fluid challenge in low (<8 ml/kg) and high tidal volume groups, the percent of correct classification of a 12% DeltaPP was 51% in the low tidal volume group and 88% in the high tidal volume group. CONCLUSIONS: DeltaPP is a reliable predictor of fluid responsiveness in mechanically ventilated patients only when tidal volume is at least 8 ml/kg. PMID- 15754197 TI - Antibiotic prophylaxis of early onset pneumonia in critically ill comatose patients. A randomized study. AB - OBJECTIVE: To evaluate if a 3-day ampicillin-sulbactam prophylaxis can reduce the occurrence of early-onset pneumonia (EOP) in comatose mechanically-ventilated patients. DESIGN: This was a single-centre, prospective, randomised, open study. SETTING: A 10-bed general-neurological ICU in a 2,000-bed university hospital. PATIENTS AND PARTICIPANTS: Comatose mechanically-ventilated patients with traumatic, surgical or medical brain injury. INTERVENTIONS: Patients were randomized to either ampicillin-sulbactam prophylaxis (3 g every 6 h for 3 days) plus standard treatment or standard treatment alone. MEASUREMENTS AND RESULTS: Main outcome was the occurrence of EOP. Secondary outcome measures were occurrence of late-onset pneumonia, percentage of non-pulmonary infections and of emerging multiresistant bacteria, duration of mechanical ventilation and of ICU stay and ICU mortality. Interim analysis at 1 year demonstrated a statistically significant reduction of EOP in the ampicillin-sulbactam group, and the study was interrupted. Overall, 39.5% of the patients developed EOP, 57.9% in the standard treatment group and 21.0% in the ampicillin-sulbactam group (chi-square 5.3971; P =0.022). Relative risk reduction of EOP in patients receiving ampicillin sulbactam prophylaxis was 64%; the number of patients to be treated to avoid one episode of EOP was three. No differences in other outcome parameters were found; however, the small sample size precluded a definite analysis. CONCLUSIONS: Antibiotic prophylaxis with ampicillin-sulbactam significantly reduced the occurrence of EOP in critically ill comatose mechanically ventilated patients. This result should encourage a large multicenter trial to demonstrate whether ampicillin-sulbactam prophylaxis reduces patient mortality, and whether antibiotic resistance is increased in patients receiving prophylaxis. PMID- 15754198 TI - Goal-directed therapy may improve outcome in complex patients--depending on the chosen treatment end point. PMID- 15754199 TI - Surface triggering receptor expressed on myeloid cells 1 expression patterns in septic shock. AB - OBJECTIVE: To analyze the pattern of cell-surface expression of the triggering receptor expressed on myeloid cells (TREM) 1 during septic shock. DESIGN AND SETTING: Prospective clinical study in an adult 16-bed medical ICU. PATIENTS AND METHODS: 25 septic shock patients, 15 patients with shock of noninfectious origin and 7 healthy volunteers. Arterial blood was drawn within 12 h of admission and subjected to flow cytometry analysis after staining with anti-TREM-1 and anti CD14 antibodies. Repeated sampling was performed on days 2, 3, 5, 7, and 14 in septic shock patients. RESULTS: Monocytic TREM-1 expression was significantly higher in septic shock patients (mean fluorescence intensity 2.3+/-0.2) than in nonseptic patients (1.0+/-0.1), and healthy volunteers (1.0+/-0.1). There was no difference in monocytic TREM-1 expression between nonseptic patients and healthy volunteers or between any of the three groups with respect to TREM-1 expression on neutrophils. The time course of TREM-1 expression on monocytes diverged significantly by day 3 between survivors and ns. CONCLUSIONS: The specificity of TREM-1 regulation by infection is highlighted. Moreover, surface TREM-1 expression on monocytes may prove useful in allowing the follow-up of septic patients during the course of the disease. PMID- 15754200 TI - Online vs live methods for teaching difficult airway management to anesthesiology residents. AB - OBJECTIVE: To compare the effectiveness of traditional and online teaching methods for educating anesthesiology residents in the principles and practice of difficult airway management. DESIGN: Prospective, randomized, controlled trial. SETTING: University hospital. PARTICIPANTS: Two randomly selected groups, each containing 28 physicians enrolled from among residents in Anesthesiology and Intensive Care Medicine in Italy. INTERVENTIONS: Residents in Group 1 took a traditional 5-h course on the principles and practice of airway management, which included lectures, slide projection, and dummy demonstrations. The same material was presented to Group 2 in an exclusively online format, which could be individually accessed for a period of 36 h. In the online course, student instructor interaction was provided through threaded discussion forums during three 30-min real-time question-and-answer sessions. MEASUREMENTS AND RESULTS: Differences in baseline and post-course scores on written tests and practical skills tests were measured. Knowledge gains in Group 2 were slightly, but not significantly, greater compared with Group 1 both in written (P=0.228) and practical skills (P=0.376) tests. Semi-quantitative ratings of learner satisfaction were significantly higher in the online group (P=0.014). Almost all online students (93%) were logged in for at least 45 of the 90 min of real-time question-and-answer sessions. The four instructors spent an average of 144+/-10 min preparing answers and interacting with online students. CONCLUSIONS: Online teaching formats may be a valid alternative for teaching residents the principles and practice of difficult airway management. Interaction with instructors seems to be an important element, but it may require substantial time commitments by instructors. PMID- 15754204 TI - The association between blood metal concentrations and heart rate variability: a cross-sectional study. AB - OBJECTIVES: Analysis of heart rate variability (HRV) is useful for measuring cardiovascular autonomic function. The effects of blood metals on cardiovascular autonomic function have not been studied extensively. The objective of the present study was to determine the association between the concentrations of blood metals, including toxic and essential trace elements, and cardiovascular autonomic function, based on HRV, in subjects without clinical cardiovascular diseases. METHODS: The subjects were public officials and their family members (n = 331) in a district of Seoul, Republic of Korea. Age, height, weight, smoking habits, medical history, blood pressure, electrocardiogram (ECG) recording, and chest X-ray were assessed by means of a self-administered questionnaire and medical examination. Blood metal concentrations (blood Pb, As, and Cd; serum Al, Co, Cu, and Zn) were measured by inductively coupled plasma-mass spectrometry. HRV parameters (low frequency, LF; high frequency, HF; total power spectrum, TPS) were measured with LRR-03 and MemCalc software (GMS, Japan). RESULTS: The concentrations of each of the blood metals were almost within normal ranges. Age and heart rate were negatively associated with LF, HF, and TPS (P< 0.01). Whole blood Cd concentrations were negatively associated with LF (P< 0.01) and HF (P< 0.05). Serum Zn concentrations were positively associated with LF and TPS (P< 0.01). CONCLUSION: Our study suggests that levels of blood metals even within normal ranges may affect heart rate variability. PMID- 15754201 TI - The histone-deacetylase inhibitor SAHA potentiates proapoptotic effects of 5 fluorouracil and irinotecan in hepatoma cells. AB - Treatment for advanced stages of hepatocellular carcinoma (HCC) remains unsatisfactory. While 5-fluorouracil (5-FU) and irinotecan are first-line treatment options for other gastrointestinal tumors, their effect on HCCs is low. Histone-deacetylase inhibitors such as suberoylanilide hydroxamic acid (SAHA) have shown antitumoral activity at micromolar concentrations in a variety of human cancers in vitro and in vivo. Here, we investigated the effects of a combination of 5-FU, irinotecan and SAHA on growth inhibition and apoptosis induction in HCC cell lines. HepG2, Hep1B and MH-7777A hepatoma cell lines and human foreskin fibroblasts as non-transformed controls were incubated with 5-FU, irinotecan and SAHA either alone or in combination. While the single agents did not show any effects on growth of the cell lines, the combination of 5-FU and irinotecan (both 10 microM) led to a moderate increase in apoptosis and proliferation inhibition. Adding 1 microM SAHA increased the apoptosis rate in hepatoma cell lines up to 92% after 72 h, while fibroblasts showed no response (5.5% apoptosis). Induction of apoptosis was paralleled by loss of the mitochondrial transmembrane potential, downregulation of bcl-2 expression and activation of caspase 3 but not caspase 8. In summary, SAHA sensitized HCC cell lines for treatment with an otherwise ineffective combination of 5-FU and irinotecan and led to mitochondrial apoptosis induction. The use of the triple combination could optimize treatment results in vivo and needs further evaluation. PMID- 15754203 TI - A phase IB study of the pharmacokinetics of gemcitabine and pemetrexed, when administered in rapid sequence to patients with advanced solid tumors. AB - BACKGROUND: We have previously demonstrated that pemetrexed is clinically active when administered 90 min after gemcitabine in a phase I study. The present study was undertaken to evaluate the efficacy, toxicity, and pharmacokinetics of gemcitabine and pemetrexed when pemetrexed is administered immediately after gemcitabine. PATIENTS AND METHODS: A total of 14 patients received 84 cycles of treatment. Gemcitabine 1250 mg/m(2) was administered on days 1 and 8 of each 21 day cycle, and pemetrexed 500 mg/m(2) on day 8 immediately following gemcitabine administration. Toxicities were graded according to the National Cancer Institute Common Toxicity Criteria and recorded as maximum grade per patient for all treatment cycles. Pharmacokinetic analyses of plasma gemcitabine and pemetrexed concentrations were performed. RESULTS: Neutropenia was the most common severe toxicity. Non-hematologic toxicities, which included nausea, vomiting, fatigue, diarrhea, rash, and elevated transaminases were of mild-to-moderate severity. No increased toxicity was observed with this schedule in comparison to the previous phase I schedule. There was no pharmacokinetic interaction between the two drugs. One partial response was documented in a patient with non-small-cell lung cancer. Eight patients had disease stabilization for five or more cycles. CONCLUSION: Gemcitabine immediately followed by pemetrexed is well tolerated and clinically active, and deserves further evaluation in phase II trials. PMID- 15754205 TI - Phase I study of TNFalpha AutoVaccIne in patients with metastatic cancer. AB - We evaluated the safety and immunogencity of a novel vaccine directed against autologous TNFalpha in a Phase I fixed dose escalation trial. The vaccine consisted of two recombinant TNFalpha proteins, with specific peptides replaced by foreign immunodominant T cell epitopes from tetanus toxoid. The main objectives were to establish a safe dose and evaluate the vaccines ability to raise neutralising TNFalpha antibodies. Secondary objectives were improvements in body weight and tumour response. Thirty-three patients were vaccinated with three doses (20, 100, or 400 mug) of TNFalpha vaccine at 2-weekly intervals adjuvanted with aluminium hydroxide. Anti-TNFalpha antibody titres were measured by both a RIA, using soluble native TNFalpha as the antigen, and by an ELISA using immobilized partly denatured TNFalpha. Eleven patients (33%) had mild grade1/2 injection site reactions at the higher doses. In 10 of 20 patients, serum antibodies recognize denatured TNFalpha in the ELISA, whereas, antibody titres against native TNFalpha in the RIA were undetectable. This suggests that the production process had partly denatured the vaccine preventing the formation of cross-reacting antibodies to native TNFalpha. In conclusion, TNFalpha vaccine was able to elicit vaccine specific antibodies. However, since the antibodies were only able to cross-react with partly denatured TNFalpha, evaluation of safety and tumour responses to the TNFalpha vaccine was compromised. PMID- 15754206 TI - The distribution of genetic parameter estimates and confidence intervals from small disconnected diallels. AB - The distributions of genetic variance components and their ratios (heritability and type-B genetic correlation) from 105 pairs of six-parent disconnected half diallels of a breeding population of loblolly pine (Pinus taeda L.) were examined. A series of simulations based on these estimates were carried out to study the coverage accuracy of confidence intervals based on the usual t-method and several other alternative methods. Genetic variance estimates fluctuated greatly from one experiment to another. Both general combining ability variance (sigma(2) (g)) and specific combining ability variance (sigma(2) (s)) had a large positive skewness. For sigma(2) (g) and sigma(2) (s), a skewness-adjusted t method proposed by Boos and Hughes-Oliver (Am Stat 54:121-128, 2000) provided better upper endpoint confidence intervals than t-intervals, whereas they were similar for the lower endpoint. Bootstrap BCa-intervals (Efron and Tibshirani, An introduction to the bootstrap. Chapman & Hall, London 436 p, 1993) and Hall's transformation methods (Zhou and Gao, Am Stat 54:100-104, 2000) had poor coverages. Coverage accuracy of Fieller's interval endpoint(J R Stat Soc Ser B 16:175-185, 1954) and t-interval endpoint were similar for both h(2) and r(B) for sample sizes n 0.6), whereas reliability was uniformly low for both measurements regarding the assessment of extension (r (s) < 0.6). Interobserver agreement was consistent across all three goniometric and two out of three visual assessments regarding the measurement of flexion (r (s) > 0.6); the interobserver reproducibility of extension, however, was uniformly low both for the visual and goniometric measurements (r (s) < 0.6). CONCLUSION: Reliability of clinical assessment of range of motion should be taken critically into consideration whilst performing classical function-related scoring systems when measuring outcome after total joint arthroplasty, since these scoring systems are strongly based on a valid and reliable assessment of range of motion. PMID- 15754229 TI - [Long-term results of the Blauth knee prosthesis--current status of hinged knee prostheses]. AB - AIM: Currently there is widespread use of unconstrained knee arthroplasty even in severe deformities of the knee. In order to ascertain the value and future of constrained knee prostheses, we have assessed the results of the Blauth knee prosthesis and compared them with the results of unconstrained knee prostheses. METHODS: In a retrospective analysis we examined 227 Blauth knee prosthesis implantations performed between 1985 and 1997. Using endpoints of prosthesis removal, infection and aseptic loosening, the 10-year survivorship analysis was evaluated. RESULTS: The analysis shows a 10-year survivorship of 90.1 % using aseptic loosening or infection as an endpoint and 96.2 % using removal as an endpoint. CONCLUSION: The first designs of constrained knee prostheses were associated with a high failure rate, while more recent designs like the St. Georg knee prosthesis and the Blauth knee prosthesis show long-term results similar to condylar designs. Since there has been a large increase of knee endoprosthesis implantations in the last years, a constrained knee prosthesis with excellent long-term results remains a valuable implant in cases of revisions and difficult knee arthroplasty. PMID- 15754230 TI - [The value of preoperative knee aspiration and arthroscopic biopsy in revision total knee arthroplasty]. AB - AIM: The accurate preoperative diagnosis of occult sepsis in endoprosthetic loosening of total knee arthroplasty is the key to successful management of revision total knee arthroplasties. The aim of this study was to evaluate the results of preoperative aspiration in comparison with preoperative arthroscopic biopsy of the synovial tissue. METHOD: From 2000 to 2004 eighty-six revision total knee arthroplasties in 86 patients were performed. 60 patients had only a knee aspiration, 15 an arthroscopic biopsy and an aspiration, 11 only arthroscopic biopsy. The results of both methods were compared with the intraoperative cultures during revision surgery. RESULTS: 69 aseptic and 17 septic knee exchange arthroplasties were performed. The preoperative aspiration of the prosthetic knee joint had a sensitivity of 68.8 %, specificity of 96.6 %, positive predictive value of 84.5 % and a negative predictive value of 92.2 %. The preoperative arthroscopic biopsy had a sensitivity of 100 %, specificity of 94.7 %, positive predictive value of 87.4 % and a negative predictive value of 100 %. CONCLUSION: Preoperative aspiration of the knee is a very helpful study for the diagnosis or exclusion of infection in a prosthetic knee joint and should be a standard procedure in the diagnosis of prosthetic loosening. If after aspiration a suspicion of infection remains, then the biopsy is an accurate procedure to diagnose or exclude periprosthetic sepsis. PMID- 15754231 TI - [Meniscus replacement with quadriceps tendon--a long-term analysis]. AB - AIM: Early osteoarthritis of the knee is a common consequence ofA'complete meniscectomy. In order to prevent degenerative changes, meniscus replacement by autogenous quadriceps tendon was performed from 1986 until 1999 in 45 patients. In 29 patients, a reconstruction of the anterior cruciate ligament was done simultaneously. This study was intended to detect whether this meniscus replacement led to better subjective, clinical and radiological results in comparison to meniscectomised knees or meniscal allografts reported in the literature. METHODS: A standardised examination of the patients (4-17 years postoperatively, average 9 years) was carried out using the KOOS questionnaire, the IKDC examination form and a weight-bearing radiograph of the knee. RESULTS: 34 patients took part in the study. KOOS subgroups showed fair results for symptoms (25-96 points, average 65), butA'bad results for sports and quality of life (0-100, average 52; QoL 6-94, average 54). Clinical and radiological examination demonstrated on average stable knee joints without effusion (IKDC group A), but X-rays showed in most cases a clear or severe osteoarthritis of the knee (IKDC group C). Clinical and radiological findings demonstrated similar results in comparison with studies investigating results after meniscectomy without meniscus replacement. Concerning subjective results, meniscus replacement with quadriceps tendon was inferior to cryopreserved meniscal allografts. However, patients after meniscal allograft transplantation also show increasing degenerative changes of the respective joint in radiological follow-up studies. CONCLUSION: In the patient group studied here with pre-existing chondromalacia of the respective knee joint compartment and preoperative anterior instability, no advantage of meniscus replacement using quadriceps tendon over the normal course after meniscectomy could be proven. Therefore, this procedure cannot be generally recommended. PMID- 15754233 TI - [Revision of failed acetabular cups with extensive structural allografts]. AB - AIM: Structural allografts are used with encouraging results for revision of failed total hip arthroplasty and in the surgery of bone tumours. The aim of the present study is to describe the clinical and radiological results achieved with structural allografts in revision of a total hip arthroplasty. MATERIAL AND METHODS: 15 patients (12 female and 3 male patients) were revised with an acetabular defect situation of type 3 A or 3 B according to the Paprosky classification. Five fresh-frozen acetabula, nine distal femora and one proximal tibia were used for acetabular reconstruction. The rigid graft fixation was performed with 2 > or = AO screws. In one case a cemented acetabular ring was implanted, four cementless cups and ten cemented polyethylene acetabular components were used. The mean follow-up was 7.9 years (1.6-11.0 years). RESULTS: A stable osseointegration of fifteen transplanted structural allografts was achieved in thirteen cases. Two allografts (one aseptic loosening, one deep infection) failed to osseointegrate. In one case the migration of a cementless cup was registered. Revision surgery of this female patient was performed successfully with a cemented reconstruction ring. The mean Harris hip score at the latest follow-up was 81.4 points (70-99 points). CONCLUSION: For reconstruction of acetabular bone stock and restoration of the bone anatomy structural allografts can be recommended. The use of cementless cups in combination with structural grafts is to be evaluated as critical. PMID- 15754232 TI - [Cemented total hip arthroplasty in Germany--an update]. AB - AIM: The results of a national survey from 1998 had shown at the time that only around 10 % of orthopaedic surgeons in Germany had strictly implemented modern cementing techniques in total hip arthroplasty (THA). The same study was repeated 5 years later to evaluate the current situation and to determine whether modern cementing techniques have become more popular. METHODS: A detailed, slightly modified questionnaire regarding cement and bone preparation, cementing techniques on acetabulum and femur, and implant types was sent to 572 German orthopaedic and trauma hospitals, as well as to visiting surgeons with an interest in THA. In total, 293 questionnaires were available for evaluation and statistical analysis. RESULTS: Palacos bone cement remained the most widely used cement (> 90 %). The mixing times given varied significantly. Vacuum mixing systems had become more popular (67.9 %). In the femur 81.8 % of the surgeons attempted to preserve cancellous bone and 57.2 % used pulsatile lavage (jet lavage). Retrograde cement application via a cement gun was done in 71.1 %. Cement restrictors were used in more than 95 %. Only two-thirds of the surgeons implemented sustained cement pressurisation and preferred a cement mantle thickness > 2mm (64 %). Only 16.9 % made multiple small acetabular keyholes and 48.6 % used jet-lavage. In 73.1 % no cement gun was used and in 68.3 % the cement was applied at high viscosity. Manual cement pressurisation was done in 58.1 %. The Muller straight stem device remained the most popular implant. For only 5 of the over 50 stem designs implanted have long-term results been published as yet. Only 10.6 % of surgeons/centres performed > 20 and almost 50 % implanted > 100 cemented THAs/year. CONCLUSION: The results of this survey demonstrated that, in comparison to 1998, the current state of cemented THA, in particular cementing technique has significantly improved. Future emphasis should be on continued surgeon education and training, as the operative, i. e., cementing techniques are of utmost importance for long-term success. PMID- 15754234 TI - [What influence doe the implant have on the perioperative morbidity following internal fixation of proximal femur fracture? Analysis of dynamic hip screw and proximal femoral nailing]. AB - RATIONALE: Proximal femur fracture is a frequent finding in elderly patients. Both the dynamic hip screw (DHS) and the proximal femur nail (PFN) are established implants. The aim of our study was to assess the perioperative morbidity in a sample of 112 patients with proximal femur fracture, operated on with either DHS or PFN. MATERIAL AND METHODS: Data of 112 consecutive patients (59 DHS, 53 PFN), which consisted of 20 variables, were obtained. Nine variables were selected, which were considered to possess a potential impact on the complication rate. These variables were type of implant, sex, age, period between trauma and surgery, ASA classification, fracture classification of the ASIF, duration of surgery, blood loss, and antibiotics. They were transformed into dichotomous data to enable univariate statistical analysis and logistic regression. RESULTS: The ASA classification only was evaluated to have a predictive value as shown by the odds ratio of 2.23 (90 % confidence interval: 1.09 - 4.56). ASA 3 or 4 patients had an expected frequency, which was 2.2-fold increased as compared to patients classified as ASA 1 or 2, to suffer from perioperative complications. Using logistic regression, again the ASA classification only was shown to have a significant impact (p = 0.066, level of significance: p < 0.1) on the perioperative morbidity. CONCLUSION: As suggested by our results, neither the type of implant nor the other variables mentioned above had a significant impact on the resulting complication rate in our study sample. The ASA classification only was found to significantly increase the probability of an adverse event. This finding should be taken into account prior to initiating therapy. PMID- 15754235 TI - [Quantification and visualization of the influence of pelvic tilt upon measurement of acetabular inclination and anteversion]. AB - AIM: The indication of acetabular inclination and anteversion not only depends on definition but also on a correctly aligned patient. Determination of anteversion and inclination according to Sven-Johannsson and Visser were simulated with 3D calculations. The influence of pelvic tilt in relation to the frontal plane was evaluated and visualized. METHOD: With 3D calculations of planar X-ray photographs for artificial hip cups the normal vector of the acetabular cup was used to calculate anteversion and inclination. RESULTS: The main axis of the projected acetabular rim is equally suited to determine the cup orientation as the normal vector of the cup plane. Pelvic tilt of about 10 degrees causes measuring errors of about 8 degrees when measured with conventionally used techniques. CONCLUSION: For the correct determination of cup orientation pelvic tilt in relation to the frontal plane has to be accounted for. PMID- 15754236 TI - [Ultrasonic Quantification of spinal configuration and postural capacity for evaluation of different muscle strengthening programs in the therapy of back pain]. AB - AIM: The aim of this study was the evaluation of the effectiveness of different muscle strengthening programs in the therapy of back pain. METHODS: 102 male longshoremen aged from 29 to 63 years with chronic back pain since > two years, matched by pain intensity and functional limitations, were randomized to three test groups (TG) and one control group (CG). The test persons carried out a program for intensified muscle strengthening over a period of six months one to two times weekly in a therapeutic practice (TG1), in a health club (TG2) and in a gymnastic group (TG3). The CG did not complete any intervention. Ultrasound topometry for recording spinal configuration and postural capacity as well as the determination of pain intensity and functional limitations were used as instruments of assessment. RESULTS: The data obtained for the CG remained virtually unchanged over the period of investigation. In contrast a decrease of pain intensity and functional limitations could be proved for all TG. Furthermore, the training programs induced both an improved postural capacity while performing the arm-raising test and an increasingly erect sagittal spinal profile. The power of effectiveness was on a homogeneous level for the medical training therapy (0.41), the fitness training (0.4) and the spinal gymnastics (0.41). CONCLUSION: The three muscle strengthening programs investigated have equally favorable effects on the parameters evaluated and are qualified as effective strategies in the therapy for chronic spinal discomforts. PMID- 15754238 TI - [Follow-up study after dynamic and static stabilisation of the lumbar spine]. AB - INTRODUCTION: The clinical indication for graf's ligamentoplasty and dorsoventral fusion may be described as the "lumbar instability syndrome". A follow-up comparison between Graf's ligamentoplasty and instrumental dorsoventral fusion in a consecutive series of 52 patients was performed. METHODS: 52 patients operated on for low back pain were recalled for a clinical and radiological review (at mean 79 months postoperatively). 26 patients underwent Graf' ligamentoplasty and 26 patients underwent dorsoventral fusion. We evaluated the surgical results and measured an objective outcome using the Oswestry Score, Low Back Outcome Score (LBOS) and Visual Analogue Scale (VAS). For radiological evaluation the Mimura and the Pathria Scores were used. RESULTS: There was no statistically significant difference between the two groups, when measured by the Oswestry Score, LBOS and VAS at the latest follow-up. The difference between the preoperative and actual VAS in each group was statistically significant (p < 0.001). Furthermore, there was no provable preoperative parameter in favour of either one of these therapies. CONCLUSION: Both methods for stabilisation methods of the lumbar spine had a comparable clinical outcome. PMID- 15754237 TI - [Effects of lumbar spinal nerve analgesia on the cardiovascular system]. AB - AIM: The frequency of cardiovascular adverse effects of lumbar paravertebral nerve root infiltration was investigated. METHOD: 117 patients with sciatic pain were included prospectively. 60 % of these suffered from known cardiovascular diseases. In 50 patients, cardiac rhythm was investigated by Holter monitoring. In 100 patients blood pressure, heart rate, respiratory rate and oxygen saturation were registered continuously from 5 minutes before to 15 minutes after the administration of a paravertebral nerve root infiltration by means of a non invasive monitoring system. RESULTS: A minor rise in systolic and diastolic blood pressure, heart rate and respiratory rate as well as a normalization of these parameters after the nerve root infiltration, were found. These findings were similar for patients with and without pre-existing cardiovascular diseases. No relevant cardiac arrhythmias could be determined. 5 of the 117 patients suffered from presyncope after the nerve root infiltration. These individuals were significantly (p = 0.002) younger than those without presyncope (32.4 +/- 9.3 vs. 55.8 +/- 14.6 years). Presyncope was more frequent during the first treatment with lumbar paravertebral nerve root infiltration in comparison to repeated application of this therapy [4/27 (14.8 %) vs. 1/90 (1.1 %), p = 0.003]. 80 % of the patients who had experienced a presyncope reported a history of similar events. CONCLUSION: According to our findings, cardiovascular monitoring for lumbar paravertebral nerve root infiltrations in the treatment of sciatica does not appear to be required. Patients with a presyncope seem to be characterized by age, first treatment and a history of (pre-)syncopes. In these cases, intravenous fluid substitution might be of help in counteracting vasovagal circulatory reactions. PMID- 15754239 TI - [Analysis of acetabular changes in Morbus Perthes disease with radiomorphometry]. AB - AIM: Decreases in depth and irregularity are typical changes of the acetabulum in patients with Perthes disease and develop secondary to the femoral head involvement. Optimal timing of therapy plays an important role to prevent these secondary changes. The present study investigates the influence of the amount of femoral head involvement and the patients' age on the acetabulum and the outcome. METHODS: 66 patients with 76 affected hip joints were included in the study. 20 hips had a conservative therapy, 22 an operative therapy (IVO). 34 hip joints underwent a conservative therapy at first, followed by an operative intervention. The radiomorphometric analyses at the time of diagnosis, pre- and postoperatively and at follow-up were performed with epiphyseal ratio, acetabular ratio and acetabulum-head ratio. At the time of diagnosis the patients were classified with the Catterall classification, at follow-up with Mose classification. RESULTS: Independent from therapy there was a correlation of the parameters with Catterall classification over the whole course, i.e., the higher the Catterall group the worse the parameter at the time of diagnosis as well as at follow-up. Operatively treated patients with Catterall IV tended towards a worse result of the epiphyseal ratio whereas they achieved better results in the other parameters compared to conservatively treated patients. Catteral I and II patients achieved good results. CONCLUSION: Operative treatment of the higher Catterall groups seems to be more effective than conservative therapy. Secondary changes of the acetabulum develop in proportion to the amount of femoral head involvement. PMID- 15754240 TI - [Correction of forearm deformities in children with multiple cartilaginous osteochondromas]. AB - AIM: Deformity of the forearm with shortening and bowing is common in children with multiple cartilaginous osteochondromas. The objective of this study was to evaluate the benefit of ulnar lengthening using an external fixateur in these patients. METHOD: 9 patients (10 cases) underwent surgery of the forearm between 1995 and 2001 and were evaluated using a standard protocol. The mean follow-up was 33.6 months, the mean age at operation 8.9 years. All patients were treated with ulnar lengthening, in 6 cases combined with an excision of the osteochondromas. RESULTS: Four out of ten patients did show an improvement in postoperative forearm rotation, two deteriorated and 4 presented unchanged. Wrist motion improved in 7 patients and remained unchanged in 3. The postoperative radial articular angle showed an improvement in 6, the carpal slip in 9 of the patients. The preoperative radial head dislocation in one patient remained unchanged postoperatively. CONCLUSION: The authors advocate this therapeutic concept for the correction of forearm deformity in multiple hereditary osteochondromas to prevent a progression of the deformity and to establish carpal stability. A significant improvement of forearm and wrist function could not be reached. PMID- 15754242 TI - [Formation of desmoids in postoperative lumbar scars--a case report]. AB - Desmoids are tumours of the connective tissue cells with aggressive growth. Frequent recurrences of these tumours have often been described. Genetic and endocrine factors as well as local tissue biomechanics are thought to be responsible for the pathogenesis. This case report should contribute to the discussion about possible pathogenetic factors. PMID- 15754241 TI - [Reliability of transcutaneous measurement of oxygen tension on the lower leg]. AB - AIM: Measurement of transcutaneous oxygen tension is increasingly used to determine the appropriate level of amputation in patients with vascular disease. The purpose of the present study was to analyze the intra- and interrater reliability of transcutaneous oxygen [tcpO (2)] measurements in a homogeneous study group. METHOD: Five investigators assessed the transcutaneous oxygen tension of both lower legs of seven persons in a fixed setting. Assessment was repeated with the same examiners and the same examinees after 24 hours. TcpO (2) was measured at the posterior aspect of the lower leg twenty centimeters below the knee joint line. The TCM 400 Monitoring System (Radiometer Medical AIS, Bronshoj, Denmark) was used. Statistical analysis of the intra- and interrater reliability was performed with the Spearman coefficient of correlation. RESULTS: An overall mean of 56.2 +/- 10.6 mmHg was found. For the first examination, a mean of 55.3 +/- 10.6 mmHg was observed, whereas for the second examination it was 57.0 +/- 10.5 mmHg. Analysis of intrarater reliability showed a coefficient of correlation of r (s) = 0.56 (p < 0.0001). For interrater reliability, we found coefficients of correlation ranging from r (s) = 0.20 (p = 0.20) to r (s) = 0.69 (p = 0.0004). CONCLUSION: Analysis of transcutaneous oxygen tension measurements performed by different investigators in a fixed setting revealed a non- homogeneous intra- and interrater reliability, which should be taken into account prior to initiating therapy. PMID- 15754243 TI - Characterization of melanin produced by a wild-type strain of Bacillus thuringiensis. AB - Bt L-7601 is a UV resistant wild-type strain, which belongs to Bacillus thuringiensis subsp. dendrolimus serotype H4a4b. It was isolated from nature, and produced a dark brown pigment during the exponential phase of growth. Bt L-7601 had the ability to produce pigment in a general nutrition-abundant medium, which had no L-tyrosine. The pigment was identified as melanin based on chemical testing, its light absorbance, and FT-IR analysis. Bt L-7601 has a strong resistance to UV light. After 30 min irradiation its survival rate was 17 times higher than that of the strain B. thuringiensis subsp. colmeri 15A3, which had no pigment. Results of the bioassays of residual insecticidal activity of Bt formulation with and without pigment produced by Bt L-7601 against larvae of Helicoverpa armigera and Spodoptera exigua after exposure to UV irradiation showed that the pigment is an excellent UV protective agent for the insecticidal proteins. PMID- 15754244 TI - Re-identification of Gluconobacter strains based on restriction analysis of 16S 23S rDNA internal transcribed spacer regions. AB - Thirty Gluconobacter strains maintained at Culture Collection NBRC were re identified at the species level on the basis of restriction analysis of 16S-23S rDNA internal transcribed spacer (ITS) regions by digestion with two restriction endonucleases MboII and Bsp1286I. The strains examined were divided into seven groups, designated as Group I and Group III-VIII, by the combination of the restriction patterns obtained with the two restriction endonucleases. Group I included seven strains, which gave "G. oxydans patterns" with the two restriction endonucleases and were re-identified as G. oxydans. Group III included 12 strains, which gave "G. frateurii patterns" and were re-identified as G. frateurii. Group IV included six strains, which gave "G. cerinus pattern" with MboII and "G. frateurii pattern" with Bsp1286I and were re-identified as G. frateurii. Group V included one strain (NBRC 3274), which gave respectively "G. frateurii pattern" and "G. cerinus pattern" and was re-identified as G. cerinus. Group VI included one strain (NBRC 3990), which gave respectively "G. oxydans pattern" and an unidentified restriction pattern and was re-identified temporarily as G. oxydans. Group VII included two strains (NBRC 3250 and NBRC 3273), which gave respectively an unidentified restriction pattern and "G. oxydans pattern." Group VIII included one strain (NBRC 3266), which gave unidentified restriction patterns. The three strains of Group VII and Group VIII were suggested to constitute new taxa by sequencing of 16S-23S rDNA ITS regions. PMID- 15754245 TI - Analysis of microbial community structure in a biofilm on membrane surface in the submerged membrane bioreactor treating domestic wastewater on the basis of respiratory quinone profiles. AB - The objective of this study was to investigate the microbial community structure of the biofouling film formed on hollow-fiber membrane surfaces in the submerged membrane bioreactor (SMBR) with a nitrification-denitrification process. In this experiment, aeration was conducted intermittently (60 min off, 90 min on) cyclic anoxic and oxic conditions in the SMBR. The dominant quinone types of biofilm on the membrane surface in an intermittently aerated SMBR were ubiquinone (UQs)-8, 10, followed by menaquinones (MKs)-8(H4), -8(H2) and -7, but those of suspended microorganisms were UQ-8, UQ-10 followed by MKs-8, -9(H4) and -6. The change in quinone profiles of biofilm on the membrane surface suggested that UQ-9, MK-7, MK 8(H2) and MK-8(H4) contributed to microbiological fouling in the intermittently aerated SMBR treating domestic wastewater. The microbial diversities of suspended microorganisms and biofilm, calculated based on the composition of all quinones, were 9.5 and 10.9, respectively. PMID- 15754247 TI - Determination of antibiotic resistance and resistance plasmids of clinical Enterococcus species. AB - To determine the antibiotic resistance pattern and resistance plasmids, we studied 23 antibiotic-resistant clinical isolates of Enterococcus spp. which caused infection in Bayindir-Ankara Hospital, Turkey. Biochemical and physiological identification tests were applied by the Vitek system and compared with the results of protein profiles by SDS-PAGE. From 23 isolates, 20 were identified as E. faecalis, 2 as E. faecium and 1 as E. gallinarum. Twenty four antibiotics belong to 10 different groups were used in susceptibility tests. Multiple antibiotic resistance was determined in 10 of 23 Enterococcus spp. Overall resistance to the used antibiotics was 47.3% and low level resistance was 16.6%. Among the isolates tested, 8.7% demonstrated high level gentamicin resistance, 17.4% demonstrated high level streptomycin resistance, and 43.5% demonstrated penicillin resistance. High level vancomycin resistant Enterococcus spp. rate was 34.8%, and 60.9% exhibited low level resistance to vancomycin and teicoplanin. They contain plasmids which varied in numbers between 1 and 11 and the plasmid sizes ranged from 2.08 to 56.15 kb. In curing experiments with acriflavine, two different plasmids were shown in different molecular sizes of 33.49 and 13.6 kb while the first determined glycopeptide and penicillin resistance, the second one determined either glycopeptide or penicillin resistance in two different E. faecalis strains. On the other hand, a 22.58 kb plasmid, determining kanamycin resistance, was detected in an E. faecium strain. After the curing experiments, an elimination of 37.17 and 44.47 kDa protein bands was shown in E. faecium EFA1 and E. faecalis EFA13 in SDS-PAGE, respectively. This survey indicates the increase of antibiotic-resistant enterococci, especially to vancomycin in our hospital isolates. PMID- 15754246 TI - Detection of methanogens and proteobacteria from a single cell of rumen ciliate protozoa. AB - Rumen ciliate-associated bacteria and methanogenic archaea were analyzed by a 16S rRNA gene retrieved from a single cell of Polyplastron multivesiculatum, Isotricha intestinalis, and Ophryoscolex purkynjei. Rumen fluid was taken from a ruminally fistulated goat to prepare a ciliate fraction. Ciliate mixtures were incubated under mixtures of antibiotics for 48 h to eliminate extracellular bacteria. Individual cells of rumen ciliates were selected under microscopic observation after fixation with ethanol. Bacterial and archaeal 16S rRNA gene sequences were retrieved from each cell of three genera of ciliate. Two archaeal sequences related to Methanobrevibacter smithii were distributed to nearly all ciliate cells tested. These two methanogenic archaea were likely to be endosymbiotic methanogens commonly carried by the rumen ciliate, although some other sequences similar to the other genera were detected. A range of proteobacteria was retrieved from cells of P. multivesiculatum. Some sequences showed similarities to the previously known endosymbiotic proteobacteria. However, there were no proteobacteria that were carried by all the ciliate cells tested. PMID- 15754248 TI - Biosorption and recycling of gold using various microorganisms. AB - In order to obtain basic information on the biosorption and recycling of gold from aqueous systems using microbial cells, the biosorption of gold by various microorganisms was investigated. Of 75 strains of microorganisms tested (25 bacteria, 19 actinomycetes, 17 fungi and 14 yeasts), high abilities of gold biosorption from a solution containing hydrogen tetrachloroaurate (III) were found in some gram-negative bacterial strains, such as Acinetobacter calcoaceticus, Erwinia herbicola, Pseudomonas aeruginosa, and P. maltophilia. Most of the gram-positive bacteria, actinomycetes, fungi and yeasts had a lower ability for gold biosorption than gram-negative bacteria. On the other hand, all of the microorganisms tested adsorbed far smaller amounts of gold from a solution containing gold dicyanoaurate (I). The biosorption of gold from a solution containing hydrogen tetrachloroaurate (III) using P. maltophilia having a high adsorbing ability for gold was very rapid and was affected by the pH of the solution, external gold concentration, and cell amounts. P. maltophilia cells immobilized with polyacrylamide gel also have a high ability for gold biosorption. The gold adsorbed on the immobilized cells is easily desorbed with 0.1 M thiourea solution. The immobilized P. maltophilia cells can be used repeatedly in biosorption-desorption cycles. PMID- 15754249 TI - Are terrestrial Ascomycetes lacking in alginolytic activity? PMID- 15754250 TI - Gluconobacter albidus (ex Kondo and Ameyama 1958) sp. nov., nom. rev., an acetic acid bacterium in the alpha-Proteobacteria. PMID- 15754259 TI - Quiz page. Giant hydroureteronephrosis. PMID- 15754262 TI - An unusual presentation of cystic fibrosis in an adult. AB - The diagnosis of cystic fibrosis (CF) generally is made within the first few years of life, although some cases will not be diagnosed until adulthood. For most patients the diagnosis is suggested by typical CF-related symptoms such as chronic respiratory infection or maldigestion. The authors describe an adult patient with newly diagnosed CF whose presenting abnormalities consisted of hypokalemia and metabolic alkalosis. These are known complications of CF but are not common presenting features that lead to the diagnosis of CF. The authors discuss their patient's presentation and review his metabolic manifestations of CF. PMID- 15754263 TI - Mast cell chymase in the ischemic kidney of severe unilateral renovascular hypertension. AB - Chymase degrades angiotensin I (AI) to form angiotensin II (AII), probably constituting a bypass of the renin-angiotensin cascade. Chymase activity increases in some vascular diseases. In the kidney, an increase in chymase activity was reported in an animal model of ischemic kidney of renovascular hypertension (RVH); however, no such evidence has been provided in humans. We treated a 64-year-old patient with severe unilateral RVH and atherosclerosis, for whom removal of the ischemic kidney was the only option. Using immunohistochemical staining, we investigated chymase activity in the removed kidney and associated artery and vein. An increase in chymase activity, together with mast cells infiltrating the interstitium, was observed where interstitial fibrosis was seen. In the renal artery, where severe atherosclerosis was seen, and also in the vein, mast cell infiltration in the adventitia was accompanied by chymase. The captopril test showed an increase in serum aldosterone level, with a concomitant increase in plasma renin activity and decrease in blood pressure. Because the decrease in blood pressure implies a decrease in circulatory AII levels, it is plausible that in this patient, chymase had a role in AII formation in the adrenal gland to stimulate aldosterone secretion. Thus, by means of captopril, AI levels increased, and chymase may have produced AII in loci tissues, which, in turn, stimulated aldosterone secretion. This is the first report of an increase in chymase activity in the interstitium of an ischemic kidney and renal artery and vein in a patient with RVH and atherosclerosis. PMID- 15754264 TI - Modification of continuous venovenous hemodiafiltration with single-pass albumin dialysate allows for removal of serum bilirubin. AB - A 53-year-old woman was admitted to the hospital with ischemic colitis and underwent a subtotal colectomy. She developed acute renal failure, severe hyperbilirubinemia, and intense pruritus resistant to medical treatment. Extracorporeal albumin dialysis using a Molecular Adsorbent Recirculating System (MARS; Gambro Co, Lund, Sweden) has been used to treat liver failure and reduce total serum bilirubin (SB) levels. A trial of extracorporeal albumin dialysis with continuous renal replacement therapy (RRT) was instituted to achieve net removal of SB. A 25% albumin solution was mixed with conventional dialysate to yield a dialysate concentration of 1.85% or 5.0% albumin. The patient underwent 2 continuous RRT sessions using extracorporeal albumin dialysis (1.85% and 5.0% albumin dialysate). Pretreatment and posttreatment SB levels were determined, and total bilirubin concentration (TB) also was measured in each of the collection bags during conventional and albumin dialysis. Pretreatment and posttreatment SB levels were 50.4 mg/dL (862 micromol/L) and 39.0 mg/dL (667 micromol/L) with 1.85% albumin dialysate and 47.1 mg/dL (805 micromol/L) and 39.7 mg/dL (679 micromol/L) with 5.0% albumin dialysate, respectively. The collected dialysate TB level was 0.3 mg/dL (5 micromol/L) during nonalbumin RRT and increased to 1.37 +/ 0.06 mg/dL (23 +/- 1 micromol/L) with 1.85% albumin dialysis. The collected dialysate fluid TB level was 0.3 mg/dL (5 micromol/L) during the nonalbumin RRT and increased to 1.38 +/- 0.15 mg/dL (24 +/- 3 micromol/L) during 5.0% albumin RRT. Single-pass albumin dialysis with continuous RRT cleared SB better than standard continuous RRT. Single-pass albumin dialysis with continuous RRT is feasible and may be a viable alternative in centers that do not have access to MARS therapy. This modality merits additional evaluation for its efficacy in clearing albumin-bound serum toxins. PMID- 15754265 TI - Treatment of fibrate-induced rhabdomyolysis with plasma exchange in ESRD. AB - Hypertriglyceridemia is a common metabolic disorder in patients with chronic renal failure. Fibrate derivatives are often used for lipid lowering in this population with adjusted dosage. However, fibrate-related adverse reaction still occasionally occurs. The authors report a case of end-stage renal disease in a patient who underwent hemodialysis regularly, taking a reduced dosage of bezafibrate (200 mg/d) for refractory hypertriglyceridemia. She did not take any statins, cyclosporine, monoamine oxidase inhibitors, or warfarin concurrently. Rhabdomyolysis was complicated along with an increased serum bezafibrate level. Plasma exchange was performed, which dramatically decreased the level of bezafibrate, and rhabdomyolysis resolved rapidly thereafter. A lower dose of bezafibrate, 200 mg every third day, was prescribed with cautious monitoring of symptoms and laboratory parameters, and better triglyceride control was achieved uneventfully. This is the first report using plasma exchange to remove excessive bezafibrate, a highly protein-bound molecule that is unlikely to be cleared by hemodialysis in an end-stage renal disease patient with serious adverse reaction caused by accumulation of bezafibrate. In contrast to a traditional wait-and-see strategy, plasma exchange seems to be a safe and effective treatment in addition to supportive care for rhabdomyolysis in such clinical scenarios. PMID- 15754266 TI - Comparison of hemodialysis, hemofiltration, and acetate-free biofiltration for ESRD: systematic review. AB - BACKGROUND: We performed a systematic review of randomized controlled trials (RCTs) comparing hemodialysis (HD), hemofiltration (HF), hemodiafiltration (HDF), and acetate-free biofiltration (AFB) in the treatment of patients with end-stage renal disease to assess their clinical effectiveness. METHODS: The Cochrane CENTRAL Registry, MEDLINE, EMBASE, CINAHL, the American College of Physicians Database, Database of Abstracts of Reviews of Effectiveness, and reference lists were searched for randomized trials of HF, HDF, and AFB compared with HD; HDF compared with AFB; and HF compared with HDF. Two reviewers extracted data for all cause mortality; hypotension, headache, nausea, vomiting, and any other adverse symptoms; quality of life (QoL); hospitalization; dialysis adequacy; and end-of treatment beta 2 -microglobulin levels. Analysis was by means of a random-effects model, and results are expressed as relative risk (RR) and weighted mean difference (WMD) with 95% confidence intervals (CIs). RESULTS: Eighteen eligible trials (588 patients) were identified. HDF was associated with significantly greater mortality risk than HD (4 trials, 326 patients; RR, 3.52; 95% CI, 1.37 to 9.47). Risk for mortality was not different among the other comparisons. Risks for hypotension episodes and dialysis-related symptoms were not significantly different with HD, HF, HDF, and AFB (18 trials, 583 patients). QoL, assessed by using an unvalidated scoring tool, appeared to be significantly improved in patients on HDF therapy than those on HD therapy (1 trial, 67 patients; WMD, 0.6; 95% CI, 0.3 to 0.9), but this was not evident when validated QoL assessment tools were used. Use of AFB compared with HDF was not associated with a significant difference in risk for hospitalization (1 trial, 11 patients; WMD, -0.45; 95% CI, -1.42 to 0.52). HDF in comparison to HD did not reduce the risk for carpal tunnel syndrome (1 trial, 67 patients; RR, 2.04; 95% CI, 0.59 to 7.00). Kt/V was significantly different with HDF compared with HD (3 trials, 124 patients; WMD, 0.14; 95% CI, 0.05 to 0.22). No other substantial data for these interventions and their impact on major patient-centered outcomes were available. CONCLUSION: The trials assessed were not powered adequately and had suboptimal method quality. It is not possible on the basis of effectiveness to prefer one extracorporeal renal replacement therapy modality to the other for end-stage kidney disease because significant differences in clinically important outcomes have not been shown by available published RCTs. PMID- 15754267 TI - Cognitive function in dialysis patients. AB - There are few detailed studies of cognitive function in dialysis patients. However, appreciating the prevalence and risk factors for cognitive impairment is important because cognitive impairment may decrease an individual's quality of life, increase resource utilization, and result in suboptimal medical care because of difficulty following caregiver recommendations. Cognitive impairment also is likely to become more of a problem as the dialysis population ages. In this review, we argue that cerebrovascular disease is an important cause of cognitive impairment in dialysis patients and discuss risk factors specific for vascular disease, as well as other factors that may influence cognitive function. We describe the structural brain abnormalities frequently seen in dialysis patients and the specific neurocognitive changes noted in prior studies. We explore potential measures to reduce cognitive impairment in this population. We conclude that additional research is needed in this area. PMID- 15754268 TI - Application of GFR-estimating equations in Chinese patients with chronic kidney disease. AB - BACKGROUND: To evaluate whether the Modification of Diet in Renal Disease (MDRD) equations could be applied accurately to Chinese patients with chronic kidney disease (CKD), glomerular filtration rates (GFRs) estimated by using MDRD equation 7 (7GFR), the abbreviated MDRD equation (aGFR), and the Cockcroft-Gault equation (cGFR) were compared in patients with different stages of CKD. METHODS: The study enrolled patients with CKD diagnosed according to the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative guidelines. All patients were older than 18 years and without acute renal function deterioration, edema, skeletal muscle atrophy, or amputation. Sex, age, body height, and body weight were recorded, and plasma creatinine levels were measured by means of Jaffe's kinetic method using a Hitachi 7600 analyzer (Hitachi, Tokyo, Japan; reagents from Roche Diagnostics, Mannheim, Germany). Creatinine, urea, and albumin were measured in a single clinical laboratory. Dual plasma sampling of technetium Tc 99m-labeled diethylene triamine pentaacetic acid plasma clearance was used as the reference standard GFR (sGFR) for comparison of 7GFRs, aGFRs, and cGFRs at different stages of CKD. RESULTS: The study enrolled 261 patients, including 146 men and 115 women. Causes of CKD included primary or secondary glomerular disease, obstructive kidney disease, chronic tubulointerstitial disease, and others. Values for 7GFR, aGFR, and cGFR were significantly greater than for sGFR in patients with CKD stages 4 to 5 (the lower the sGFR, the greater the difference); whereas 7GFR, aGFR, and cGFR were significantly lower than sGFR in patients with CKD stage 1. CONCLUSION: Our results show that in a Chinese population with CKD, MDRD equation 7 and the abbreviated MDRD equation overestimated GFR in patients with CKD stages 4 to 5 and underestimated GFR in those with CKD stage 1. These results indicate that careful modification of these equations may be necessary in Chinese populations with CKD. PMID- 15754269 TI - First United Kingdom Heart and Renal Protection (UK-HARP-I) study: biochemical efficacy and safety of simvastatin and safety of low-dose aspirin in chronic kidney disease. AB - BACKGROUND: Patients with chronic kidney disease are at increased risk for cardiovascular disease, but the efficacy and safety of simvastatin and aspirin are unknown in this patient group. METHODS: Patients were randomly assigned in a 2 x 2 factorial design to the administration of: (1) 20 mg of simvastatin daily versus matching placebo, and (2) 100 mg of modified-release aspirin daily versus matching placebo. RESULTS: Overall, 448 patients with chronic kidney disease were randomly assigned (242 predialysis patients with a creatinine level > or = 1.7 mg/dL [> or =150 micromol/L], 73 patients on dialysis therapy, and 133 patients with a functioning transplant). Compliance with study treatments was 80% at 12 months. Allocation to treatment with 100 mg of aspirin daily was not associated with an excess of major bleeds (aspirin, 4 of 225 patients [2%] versus placebo, 6 of 223 patients [3%]; P = not significant [NS]), although there was a 3-fold excess of minor bleeds (34 of 225 [15%] versus 12 of 223 patients [5%]; P = 0.001). Among those with predialysis renal failure or a functioning transplant at baseline, aspirin did not increase the number of patients who progressed to dialysis therapy (7 of 187 [4%] versus 6 of 188 patients [3%]; P = NS) or experienced a greater than 20% increase in creatinine level (63 of 187 patients [34%] versus 56 of 188 patients [30%]; P = NS). After 12 months of follow-up, allocation to 20 mg of simvastatin daily reduced nonfasting total cholesterol levels by 18% (simvastatin, 163 mg/dL [4.22 mmol/L] versus placebo, 196 mg/dL [5.08 mmol/L]; P < 0.0001), directly measured low-density lipoprotein cholesterol levels by 24% (89 mg/dL [2.31 mmol/L] versus 114 mg/dL [2.96 mmol/L]; P < 0.0001), and triglyceride levels by 13% (166 mg/dL [1.87 mmol/L] versus 186 mg/dL [2.10 mmol/L]; P < 0.01), but there was no significant effect on high-density lipoprotein cholesterol levels (2% increase; P = NS). Allocation to simvastatin therapy was not associated with excess risk for abnormal liver function test results or elevated creatine kinase levels. CONCLUSION: During a 1-year treatment period, simvastatin, 20 mg/d, produced a sustained reduction of approximately one quarter in low-density lipoprotein cholesterol levels, with no evidence of toxicity, and aspirin, 100 mg/d, did not substantially increase the risk for a major bleeding episode. Much larger trials are now needed to assess whether these treatments can prevent vascular events. PMID- 15754270 TI - Fenofibrate reduces progression to microalbuminuria over 3 years in a placebo controlled study in type 2 diabetes: results from the Diabetes Atherosclerosis Intervention Study (DAIS). AB - BACKGROUND: Microalbuminuria is an early marker of diabetic nephropathy and an independent risk factor for cardiovascular disease. In the Diabetes Atherosclerosis Intervention Study (DAIS), treatment of people with type 2 diabetes with micronized fenofibrate for an average of 38 months reduced the progression of angiographically evaluated coronary artery disease and improved lipoprotein level abnormalities compared with placebo. The aim of this analysis is to study the influence of the treatment on changes in urinary albumin excretion. METHODS: Microalbuminuria was measured on 2 to 3 occasions by using timed overnight samples at baseline and yearly thereafter in 314 DAIS participants (77 women, 237 men; average age, 56 years); all except 3 participants had either a normal albumin excretion rate (<20 microg/min; n = 214) or microalbuminuria (albumin, 20 to 200 microg/min; n = 97) before randomization. Tabulated shifts (between normal, microalbuminuria, and macroalbuminuria) from baseline to last observed values were compared between treatment groups by means of chi-square or Fisher's exact test. RESULTS: Fenofibrate significantly reduced the worsening of albumin excretion (fenofibrate, 8% versus placebo, 18%; P < 0.05). This effect was caused mostly by reduced progression from normal albumin excretion to microalbuminuria: 3 of 101 participants in the fenofibrate group versus 20 of 113 participants in the placebo group (P < 0.001). Overall, changes in albumin excretion were independent of age or changes in lipid or creatinine levels, weight, or blood pressure. CONCLUSION: Improvement in lipid profiles with fenofibrate in patients with type 2 diabetes was associated with reduced progression from normal albumin excretion to microalbuminuria. PMID- 15754271 TI - Stepwise increase in arterial stiffness corresponding with the stages of chronic kidney disease. AB - BACKGROUND: Patients with end-stage renal disease on maintenance dialysis therapy have a high prevalence of cardiovascular risk factors and cardiovascular disease (CVD). A similar finding is noted in patients with chronic kidney disease (CKD). The important contributors are premature and accelerated atherosclerosis and vascular calcification. We assessed the severity of arterial stiffness in 102 patients with CKD by using pulse wave velocity (PWV) and sought to identify associated risk factors. METHODS: PWV was measured by calculating the distance traveled by the flow wave and divided by the time delay. Correlations between PWV and traditional cardiovascular risk factors, estimated glomerular filtration rate (GFR) per 1.73 m2 , blood pressure (BP), and pulse pressure (PP) were analyzed. RESULTS: PWV values in patients with CKD stages 1 to 2 and the age-matched control group were similar. There was a significant trend for a stepwise increase in PWV corresponding to advance in CKD stage (P < 0.0001). Univariate linear regression analysis showed that age, prior CVD, diabetes, hypertension, any high risk, estimated GFR per 1.73 m2 , systolic BP, and PP correlated with PWV. In the multivariate model, decreased estimated GFR per 1.73 m2 and increased systolic BP were independently associated with increased PWV in patients with CKD (model R 2 = 0.539; P < 0.0001). CONCLUSION: This is the first study to show a greater PWV in patients with more advanced CKD from stages 1 to 5. Estimated GFR per 1.73 m2 and systolic BP were the major clinical determinants of arterial stiffness in patients with CKD independent of conventional risk factors for CVD. PMID- 15754273 TI - Myeloid-related protein 8 expression on macrophages is a useful prognostic marker for renal dysfunction in children with MPGN type 1. AB - BACKGROUND: To clarify the role of subclasses of macrophages in chronic glomerulonephritis, we evaluated the relationship between myeloid-related protein 8 (MRP8) and MRP14 expression on macrophages and the progression of membranoproliferative glomerulonephritis (MPGN). METHODS: We enrolled 35 patients with MPGN type 1 who had a normal creatinine clearance at the time of their first renal biopsy and divided them into 2 groups based on clinical status at the time of their most recent examination: 12 patients with normal urine test results and 12 patients with minor urinary abnormalities at the latest observation (group 1) and 7 patients with persistent nephropathy and 4 patients with renal insufficiency (group 2). The first and second renal biopsy findings and MRP8 and MRP14 expression on macrophages were investigated in both groups. RESULTS: Mean scores for positive glomerular and interstitial MRP8 and CD68 staining at the time of the first and second biopsies were significantly higher in group 2 than group 1. At the time of the second biopsy, mean scores for interstitial CD68 positive (CD68 +) staining were higher in group 2 than group 1. Mean scores for glomerular and interstitial MRP8 + and CD68 + staining at the time of the first biopsy correlated with the chronicity index at the time of second biopsy in both groups. CONCLUSION: Results suggest that MRP8 expression on macrophages in glomeruli and interstitial lesions at the first biopsy can be a useful prognostic marker for renal dysfunction in children with MPGN type 1. PMID- 15754272 TI - Comparison of ARF after myeloablative and nonmyeloablative hematopoietic cell transplantation. AB - BACKGROUND: Acute renal failure (ARF) occurs with significant frequency after myeloablative and nonmyeloablative allogeneic hematopoietic cell transplantation (HCT). Myeloablative (conventional) HCT is the standard of care for cure of various malignant disorders. The newer modality of nonmyeloablative ("mini-allo") HCT is reserved for patients with advanced age and comorbidities who are ineligible for myeloablative HCT. The present study compares the incidence of ARF between patients undergoing concurrent myeloablative and nonmyeloablative HCT in the same period at the same institution. METHODS: This retrospective cohort study from 1997 to 2003 compares 140 myeloablative and 129 nonmyeloablative patients from the Fred Hutchinson Cancer Research Center. Severity of ARF was classified into 4 grades based on the increase in serum creatinine levels in the first 100 days after HCT. Mortality was studied at 100 days and 1 year. RESULTS: Nonmyeloablative patients were significantly older and had greater pretransplantation comorbidity at baseline. Despite this, patients undergoing myeloablative HCT had a greater incidence of severe ARF (grades 2 and 3, 73% versus 47%; P < 0.001). The incidence of dialysis also was 4-fold greater (12% versus 3%; P < 0.001) in the myeloablative than nonmyeloablative group. On multivariate analysis after controlling for baseline characteristics, myeloablative HCT was associated with a 4.8-fold greater incidence of ARF compared with nonmyeloablative HCT. Nonrelapse mortality also was greater in the myeloablative group at 100 days and 1 year. CONCLUSION: The incidence and severity of ARF, as well as nonrelapse mortality, occurring after nonmyeloablative HCT is significantly lower compared with myeloablative HCT. PMID- 15754274 TI - Association of genetic polymorphisms with risk of renal injury after coronary bypass graft surgery. AB - BACKGROUND: Post-cardiac surgery renal dysfunction is a common, serious, multifactorial disorder, with interpatient variability predicted poorly by preoperative clinical, procedural, and biological markers. Therefore, we tested the hypothesis that selected gene variants are associated with acute renal injury, reflected by a serum creatinine level increase after cardiac surgery. METHODS: One thousand six hundred seventy-one patients undergoing aortocoronary surgery were studied. Clinical covariates were recorded. DNA was isolated from preoperative blood; mass spectrometry was used for genotype analysis. A model was developed relating clinical and genetic factors to postoperative acute renal injury. RESULTS: A race effect was found; therefore, Caucasians and African Americans were analyzed separately. Overall, clinical factors alone account poorly for postoperative renal injury, although more so in African Americans than Caucasians. When 12 candidate polymorphisms were assessed, 2 alleles (interleukin 6 -572C and angiotensinogen 842C) showed a strong association with renal injury in Caucasians (P < 0.0001; >50% decrease in renal filtration when they present together). Using less stringent criteria for significance (0.01 > P > 0.001), 4 additional polymorphisms are identified (apolipoproteinE 448C [4], angiotensin receptor1 1166C, and endothelial nitric oxide synthase [eNOS] 894T in Caucasians; eNOS 894T and angiotensin-converting enzyme deletion and insertion in African Americans). Adding genetic to clinical factors resulted in the best model, with overall ability to explain renal injury increasing approximately 4-fold in Caucasians and doubling in African Americans (P < 0.0005). CONCLUSION: In this study, we identify genetic polymorphisms that collectively provide 2- to 4-fold improvement over preoperative clinical factors alone in explaining post-cardiac surgery renal dysfunction. From a mechanistic perspective, most identified genetic variants are associated with increased renal inflammatory and/or vasoconstrictor responses. PMID- 15754275 TI - Nonsteroidal anti-inflammatory drugs and risk of ARF in the general population. AB - BACKGROUND: Most epidemiological studies evaluating the association between nonsteroidal anti-inflammatory drugs (NSAIDs) and acute renal failure (ARF) found an increased risk for developing ARF while taking NSAIDs. Despite these studies, little is known about the effect of dose and duration of therapy, risk of individual NSAIDs, comorbidity, or concomitant use of other nephrotoxic drugs. METHODS: This is a nested case-control study using the General Practice Research Database from the United Kingdom. Participants were 386,916 patients aged 50 to 84 years on January 1, 1997, and free of known cancer, renal disorder, cirrhosis, or systemic connective tissue disease. After validation of cases identified from this cohort, 103 patients were confirmed as idiopathic cases of ARF and compared with 5,000 controls frequency matched by age and sex. RESULTS: Current users of NSAIDs had a relative risk (RR) for ARF of 3.2 (95% confidence interval [CI], 1.8 to 5.8), and the risk declined after treatment was discontinued. Increased risk was present with both short- and long-term therapy and was slightly greater among users of high doses. History of heart failure (HF), hypertension, diabetes, and hospitalizations and consultant visits in the previous year were all associated with a greater risk for ARF. There was a suggestion of a modification of the effect of NSAIDs in patients with hypertension and those with HF. Use of selected cardiovascular drugs was associated with a 5-fold increase in risk for ARF. Diuretics presented the greatest risk. Risk increased with concomitant use of NSAIDs and diuretics (RR, 11.6; 95% CI, 4.2 to 32.2) and NSAIDs and calcium channel blockers (RR, 7.8; 95% CI, 3.0 to 20.5). CONCLUSION: NSAID users had a 3 fold greater risk for developing a first-ever diagnosis of clinical ARF compared with non-NSAID users in the general population. NSAIDs should be used with special caution in patients with hypertension and/or HF. PMID- 15754276 TI - Long-term, low-dose, intravenous vitamin C leads to plasma calcium oxalate supersaturation in hemodialysis patients. AB - BACKGROUND: Ascorbate supplementation for patients on regular dialysis treatment (RDT) is advised to obviate deficiency and improve epoetin response in those with functional iron deficiency. However, clear-cut safety concerns regarding hyperoxalemia are still poorly understood. This study tries to establish safety/efficacy profiles of ascorbate and oxalate during long-term intravenous ascorbate supplementation. METHODS: A prospective study was performed in 30 patients on RDT showing ascorbate deficiency (plasma ascorbate < 2.6 mg/L [<15 micromol/L]): 18 patients were administered intravenous ascorbate during 18 months (250 mg/wk, subsequently increased to 500 mg), and 12 patients were taken as reference untreated cases. Plasma ascorbate and oxalate assays and dialytic balance determinations were performed (ion chromatography and reverse-phase high performance liquid chromatography, respectively) at baseline, during treatment, and 12 months after withdrawal. RESULTS: Plasma ascorbate levels increased dose dependently with supplementation (1.6 +/- 0.8 mg/L [9.1 +/- 4.6 mumol/L] at baseline, 2.8 +/- 1.8 mg/L [15.9 +/- 10.1 micromol/L]) with 250 mg of ascorbate, and 6.6 +/- 2.8 mg/L [37.5 +/- 16.0 micromol/L] with 500 mg/wk of ascorbate), but only normalized with greater dosages for several months in 94% of patients. Baseline plasma oxalate levels increased from 3.2 +/- 0.8 mg/L (35.8 +/- 8.8 micromol/L) to 3.6 +/- 0.8 mg/L (39.5 +/- 9.1 micromol/L) and 4.5 +/- 0.9 mg/L (50.3 +/- 10.4 micromol/L) with 250 and 500 mg, respectively ( P < 0.001). The calcium oxalate saturation threshold was exceeded by 7 of 18 patients (40%) during 6 months therapy with 500 mg/wk. Ascorbate dialysis removal increased from 37.8 +/- 23.2 mg (215 +/- 132 micromol) to 99.6 +/- 51.7 mg (566 +/- 294 micromol) during supplementation (P < 0.001), with corresponding increases in oxalate removal from 82.5 +/- 33.2 mg (917 +/- 369 micromol) to 111.2 +/- 32.6 mg/L (1,236 +/- 362 micromol; P < 0.01). Withdrawal reverted plasma levels and dialysis removal to initial values. Values for untreated patients did not change during 1 year of follow-up. CONCLUSION: Patients on RDT may resolve ascorbate deficiency with intravenous supplementation of 500 mg/wk, but this implies a significant risk for oxalate supersaturation. Oxalate measurements are strongly recommended during long-term ascorbate therapy. PMID- 15754277 TI - Association of conjunctival and corneal calcification with vascular calcification in dialysis patients. AB - BACKGROUND: Conjunctival and corneal calcification (CCC) is a well-known and easily detectable extraskeletal calcification, but its association with vascular calcification was not investigated previously. The aim of this study is to investigate the relationship of CCC with vascular calcification and bone metabolism parameters in dialysis patients. METHODS: We evaluated 63 patients (30 men, 33 women; mean age, 43.5 +/- 13.4 years) who were on dialysis therapy for more than 6 months. Forty-four patients were on peritoneal dialysis and 19 patients were on hemodialysis therapy. The same observer evaluated the presence of CCC by using a slit-lamp microscope, and a total CCC score was recorded for each patient. Fifty-two age- and sex-matched healthy controls also were evaluated by using the same method. Biochemical data were collected from patient files. Bone mineral density (BMD) of the lumbar spine and femoral neck was measured, and the presence of vascular calcification was assessed by using x-ray examinations of the pelvis and hands. RESULTS: Mean CCC score in patients was significantly higher than that in controls (6.2 +/- 5.1 versus 1.3 +/- 1.8; P = 0.001). CCC score correlated significantly with duration of renal replacement therapy ( r s = 0.392; P = 0.002), serum phosphorus level ( r s = 0.259; P = 0.042), and calcium x phosphorus product ( r s = 0.337; P = 0.007). However, we did not find a significant correlation with calcium, parathyroid hormone, alkaline phosphatase, albumin, or C-reactive protein level or BMD. The frequency of vascular calcification was significantly greater in patients with a high CCC score (CCC score > or = 10) compared with a low CCC score (< or =3; 56.3% versus 5.6%; P = 0.002). CONCLUSION: Evaluation of CCC score is an easy, fast, and noninvasive method. It seems that CCC score can be used as an additional tool to assess the status of extraskeletal calcification in dialysis patients. PMID- 15754278 TI - Citrate for long-term hemodialysis: prospective study of 1,009 consecutive high flux treatments in 59 patients. AB - BACKGROUND: Regional citrate anticoagulation during hemodialysis is performed in selected patients at highly specialized units. We postulated that routine use of citrate at a long-term dialysis ward is safe and efficient. METHODS: During a 2 year period, we studied 1,009 consecutive citrate-anticoagulated high-flux hemodialysis treatments performed in 59 patients at our long-term dialysis ward. We used a simple citrate infusion protocol, calcium-free dialysate, and intravenous calcium substitution. Simple and clear algorithms allowed adjustments of the calcium substitution rate and dialysate settings by the attending nurse. Adverse events; indications for citrate anticoagulation; clotting; technical data; blood ionized calcium, sodium, and potassium levels; and acid-base homeostasis were analyzed prospectively. RESULTS: Of the treatments, 99.6% were accomplished successfully. Two adverse events were attributed to citrate use. Overall, ionized calcium levels were stable during the procedures and electrolyte and acid-base balances were well controlled. The use of central venous catheters for dialysis was associated with paradoxical behavior of ionized calcium levels (increasing blood ionized calcium levels despite decreased calcium infusion). Anticoagulation was excellent. CONCLUSION: Routine use of citrate anticoagulation in the setting of a long-term hemodialysis ward is safe and efficient. Measured ionized calcium levels should be interpreted with care if central venous catheters are used for vascular access because they could be biased by recirculation. PMID- 15754280 TI - The Renal Treatment Satisfaction Questionnaire (RTSQ): a measure of satisfaction with treatment for chronic kidney failure. AB - BACKGROUND: Quality and effectiveness of care can be enhanced through the use of condition-specific measures of satisfaction with treatment. The aim of the present study was to design and develop a measure of satisfaction with treatment for patients with chronic kidney failure (CKF) for use in routine clinical care and clinical trials. The Renal Treatment Satisfaction Questionnaire (RTSQ) was designed to be suitable for people using any of the various treatment modalities for CKF. Items measure satisfaction with aspects of treatment, including convenience, flexibility, freedom, and satisfaction to continue with present form of treatment. METHODS: A 12-item RTSQ was investigated at a UK hospital-based renal unit, using data from 140 outpatients undergoing renal replacement therapy (hemodialysis, n = 35; continuous ambulatory peritoneal dialysis, n = 57; transplantation, n = 46). RESULTS: An 11-item scale was developed from the original 12-item version, with a single factor accounting for 59% of the variance and item loadings greater than 0.58. Scale reliability was excellent (alpha = 0.93) in the full sample and proved robust to analysis in separate treatment subgroups. As expected, RTSQ scores differed significantly ( P < 0.0001) between the transplantation and other treatment groups. Those who had received a transplant expressed greater overall satisfaction, with specific advantages of transplantation shown by all individual items, including convenience, time, lifestyle, freedom, and satisfaction to continue current treatment. CONCLUSION: The RTSQ provides a brief reliable measure of satisfaction with treatment for patients with CKF that is suitable for use in routine clinical care and clinical trials. PMID- 15754279 TI - Dialyzer membrane permeability and survival in hemodialysis patients. AB - BACKGROUND: We previously showed that nutritional protein concentrations were predictive of outcome, whereas variables reflecting body composition and dialysis dose were not, in a 30-month prospective follow-up of 1,610 hemodialysis patients. Information on dialysis membrane and erythropoietin use had to be evaluated in an additional follow-up. METHODS: A subset of 650 patients from the initial cohort of 1,610 was analyzed for survival in a 2-year extension of follow up. Detailed data were collected: demographics; cause of renal failure; time on dialysis therapy; type of membrane; erythropoietin treatment; body mass index (BMI); predialysis albumin, prealbumin, and bicarbonate levels; and outcome. Normalized protein catabolic rate (nPCR), dialysis adequacy, and lean body mass were computed from predialysis and postdialysis urea and creatinine values. RESULTS: Patient characteristics were age of 61 +/- 16 years, 58% men, BMI of 22.7 +/- 4.4 kg/m2 , time on dialysis therapy of 102 +/- 73 months, and 8.8% had diabetes. Dialysis parameters were duration of 247 +/- 31 minutes, Kt/V of 1.4 +/ 0.3, and nPCR of 1.2 +/- 0.3 g/kg/d. Albumin level was 3.73 +/- 0.53 g/dL (37.3 +/- 5.3 g/L), and prealbumin level was 31 +/- 8 mg/dL. The survival rate was 78.7% after 2 years. Survival was influenced by age, presence of diabetes, use of high-flux membrane, and serum albumin level, but not other variables, including Kt/V and prealbumin level. Two-year variations in values for urea, creatinine, and weight were predictive of survival in univariate, but not multivariate, analyses. CONCLUSION: In patients on dialysis therapy for a long period, better survival was observed when high-flux dialysis membranes were used. PMID- 15754281 TI - A case of renal sarcoidosis with complement activation via the lectin pathway. AB - A 57-year-old woman with pulmonary sarcoidosis was admitted to the hospital because of an elevation of serum creatinine and blood urea nitrogen. On admission, the laboratory data suggested interstitial nephritis without proteinuria and hematuria, whereas a renal biopsy showed granulomatous interstitial nephritis and mild mesangial proliferative glomerulonephritis. Immunoglobulin and C1q deposits were negative, but mannose-binding lectin, C3, C4d, and C5b-9 deposits were marked in the glomerular mesangial areas. The lectin pathway of complement activation may have contributed to the development of glomerular injury in this patient. DNA of Propionibacterium acnes , which is now strongly suspected as the pathogen of sarcoidosis, was detected in the patient's glomerular mesangial cells; tubular epithelial cells, which were involved in granulomatous inflammation; and mononuclear cells in epithelioid granulomas by in situ hybridization. These findings may add new insights to the pathogenesis of renal sarcoidosis, including its relation to infection, because mannose-binding lectin plays a crucial role in the host defense against various pathogens. From this case of renal sarcoidosis, it is hypothesized that P acnes may be involved in pathogenesis of granulomatous interstitial nephritis and that it plays a role in glomerular complement activation via the lectin pathway. PMID- 15754282 TI - Late-onset thrombocytic microangiopathy caused by cblC disease: association with a factor H mutation. AB - BACKGROUND: cblC disease is a cause of hemolytic uremic syndrome (HUS), which has been primarily described in neonates and infants with severe renal and neurological lesions. PATIENTS: Two sisters aged 6 and 8.5 years presented with a latent hemolytic process characterized by undetectable or low plasma haptoglobin, respectively, associated with renal failure and gross proteinuria. Renal biopsies performed in both patients found typical findings of thrombotic microangiopathy suggesting the diagnosis of HUS. Both patients were free of neurologic signs. RESULTS: Biochemical investigations found a cobalamin processing deficiency of the cblC type. Search for additional factors susceptible to worsen endothelial damage revealed homozygosity 677C--> T mutation in the methylenetetrahydrofolate reductase gene as well as heterozygosity for a 3254T--> C mutation in factor H in the patient with the most severe clinical presentation. Long-term subcutaneous administration of hydroxocobalamin in combination with oral betaine and folic acid resulted in clinical and biological improvement in both patients. CONCLUSION: cblC disease may be a cause of chronic HUS with delayed onset in childhood. Superimposed mutation of factor H gene might influence clinical severity. PMID- 15754283 TI - Carnitine palmitoyltransferase II deficiency due to a novel gene variant in a patient with rhabdomyolysis and ARF. AB - Adult patients deficient in carnitine palmitoyltransferase II (CPT II) cannot generate sufficient amounts of energy, which results in rhabdomyolysis and acute renal failure (ARF). Its genetic basis has been recognized; but histopathologic changes, especially electron microscopic changes, have scarcely been described. The study subject is a patient with ARF caused by repetitive nontraumatic rhabdomyolysis. The acylcarnitine profile of serum and enzyme assay on skin fibroblasts confirmed the diagnosis of CPT II deficiency. Renal biopsy specimens were examined microscopically and immunohistochemically. The histological diagnosis was interstitial nephritis with acute tubular necrosis caused by rhabdomyolysis. Myoglobin in tubules was detected by means of immunohistochemistry and electron microscopy. The genetic structure of CPT II was analyzed in the patient and his family. Eight pairs of polymerase chain reaction (PCR) primers were designed to cover the coding region. Each PCR-amplified gene product was subjected to DNA sequencing, which unveiled heterozygosity at the CPT II locus consisting of a deletion of cytosine and thymine at codon 408, resulting in a stop signal at 420, as well as a mutation of arginine to cysteine at codon 631. The frame shift at 408 has never been described before. DNA sequencing of the family showed the deletion mutation from the mother and the point mutation from the father. We describe renopathological findings in a patient with CPT II deficiency associated with rhabdomyolysis, which suggested the pathological role of myoglobin casts in the development of tubular necrosis. Genetic analysis of the patient identified a novel variant of the CPT II gene. PMID- 15754284 TI - Birth and agony of hemofiltration. PMID- 15754286 TI - Estimating GFR from serum creatinine concentration: pitfalls of GFR-estimating equations. PMID- 15754285 TI - Statins and aspirin in chronic kidney disease: what does the UK-HARP-I trial tell us? PMID- 15754287 TI - Acute renal failure. PMID- 15754288 TI - Multiple complications in multiple myeloma. PMID- 15754289 TI - Teaching of pathology at more than one level. PMID- 15754290 TI - Pathology data integration with eXtensible Markup Language. AB - It is impossible to overstate the importance of XML (eXtensible Markup Language) as a data organization tool. With XML, pathologists can annotate all of their data (clinical and anatomic) in a format that can transform every pathology report into a database, without compromising narrative structure. The purpose of this manuscript is to provide an overview of XML for pathologists. Examples will demonstrate how pathologists can use XML to annotate individual data elements and to structure reports in a common format that can be merged with other XML files or queried using standard XML tools. This manuscript gives pathologists a glimpse into how XML allows pathology data to be linked to other types of biomedical data and reduces our dependence on centralized proprietary databases. PMID- 15754291 TI - Strategies for teaching pathology to graduate students and allied health professionals. AB - Pathology is an essential course for many students in the biomedical sciences and allied health professions. These students learn the language of pathology and medicine, develop an appreciation for mechanisms of disease, and understand the close relationship between basic research and clinical medicine. We have developed 3 pathology courses to meet the needs of our undergraduates, graduate students, and allied health professionals. Through experience, we have settled on an approach to teaching pathology that takes into account the diverse educational backgrounds of these students. Educational resources such as assigned reading, online homework, lectures, and review sessions are carefully balanced to adjust course difficulty. Common features of our pathology curricula include a web-based computer laboratory and review sessions on the basis of selected pathology images and open-ended study questions. Lectures, computer-guided homework, and review sessions provide the core educational content for undergraduates. Graduate students, using the same computer program and review material, rely more heavily on assigned reading for core educational content. Our experience adapting a pathology curriculum to the needs of divergent groups of students suggests a general strategy for monitoring course difficulty. We hypothesize that course difficulty is proportional to the information density of specific learning resources (eg, lecture or textbook) multiplied by the weight of those learning resources placed on examinations. This formula allows educators to match the difficulty of a course with the educational needs of students, and provides a useful tool for longitudinal studies of curriculum reform. PMID- 15754292 TI - Microglandular hyperplasia: a model for the de novo emergence and evolution of endocervical reserve cells. AB - BACKGROUND: Microglandular hyperplasia (MGH) of the cervix in human beings is associated early with gland proliferation and terminates in mature squamous metaplasia. Using antibodies to basal cell markers, we analyzed biopsies with MGH to profile the distribution and evolution of reserve cells and their relationship to these epithelial components. DESIGN: Serial sections of 24 MGHs were subdivided into (1) early MGH with microacinar proliferation, abundant subnuclear vacuoles, and a paucity of supporting stroma and (2) late MGH with more prominent supporting stroma and/or squamous metaplasia. Serial sections were stained with antibodies to p63, bcl-2, and keratin-5. RESULTS: Three patterns of p63 staining were observed corresponding to the age of the MGH: (1) scattered staining of columnar cells, (2) focal subcolumnar staining in a reserve cell distribution, and (3) linear subcolumnar arrays of p63-positive reserve cells that in some MGHs expanded into a squamous metaplasia. Early acinar proliferations showed weak and focal columnar cell staining followed by focal subcolumnar p63-positive cells. In late lesions, p63 staining was compartmentalized to the extraglandular (or subcolumnar) areas. Stainings of p63, bcl-2, and keratin-5 were concordant. Staining for keratin 14, which localizes to squamous cells, was variable. CONCLUSIONS: The immunohistochemical profile in MGH indicates that reserve cells are created in adulthood during specialized columnar proliferations. This columnar to reserve cell transition may produce a stable population of reserve cells or a transition to squamous metaplasia. Similar patterns are seen in cervical neoplasia, suggesting a link between benign and neoplastic cervical epithelial differentiation. PMID- 15754293 TI - Increased expression of claudins in cervical squamous intraepithelial neoplasia and invasive carcinoma. AB - Claudins (CLDNs), of which 24 types have been identified, are integral transmembrane proteins of the tight junctions that are critical for maintaining cell adhesion and polarity. They also act as selective barriers. Cells and tissues are characterized by individual CLDN patterns; the composition and levels of expression change during differentiation and tumor formation. Alterations in the expression of individual CLDNs have been detected in several carcinomas and shown to be related to progression and invasion; however, their role in carcinogenesis is controversial. Using a panel of polyclonal (CLDNs 1, 3, and 7) and monoclonal (CLDNs 2 and 4) antibodies, CLDN pattern and expression were studied by immunohistochemistry in 105 cervical tissue specimens, including normal epithelia (n = 20), cervical intraepithelial neoplasias (CINs; CIN 1/2, n = 27; CIN 3, n = 10), carcinoma in situ (CIS, n = 15), and 33 squamous keratinizing and nonkeratinizing invasive carcinomas. No CLDN 3 was observed in normal or intraepithelial neoplastic cells, but significantly increased expression of CLDNs 1, 2, 4, and 7 was detected in the CIN/CIS lesions and invasive carcinomas compared with the normal tissues (P < .001) and reduced reactivity of CLDNs 1 and 2 was observed in invasive cervical cancers compared with CIN 3/CIS (P = .0001) and of CLDNs 2, 4, and 7 compared with CIN 1/2. These results indicate increased expression of CLDNs in the early phase of carcinogenesis in intraepithelial lesions, which decreases during progression to invasive disease. Expression of CLDN 1 was strongest in premalignant stages; thus, it may serve as a good diagnostic marker for the detection of CIN. PMID- 15754294 TI - Guanylyl cyclase C is a marker of intestinal metaplasia, dysplasia, and adenocarcinoma of the gastrointestinal tract. AB - Gastrointestinal (GI) tumors continue to be major causes of cancer-related mortality, in part, reflecting metastases that escape detection by histopathology. Moreover, although approximately 10% of carcinomas arise from unknown locations, these tumors frequently originate in the GI tract. Guanylyl cyclase C (GC-C) is a receptor selectively expressed by intestinal epithelial cells whose persistent expression by colorectal carcinomas and ectopic expression by adenocarcinomas of the upper GI tract suggest its use as a marker for GI malignancies. Here, expression of GC-C protein, identified by immunohistochemistry, was examined in tissues and tumors arising from the human GI tract. Guanylyl cyclase C protein was expressed by epithelial cells from the duodenum to the rectum, but not by those in normal esophagus and stomach. Expression was retained in tubular adenomas, inflammatory bowel disease, premalignant lesions, and in primary and metastatic adenocarcinomas from the colon, including metastases to lymph nodes and liver. Moreover, GC-C was ectopically expressed in all cases of dysplasia and adenocarcinomas arising from intestinal metaplasia in esophagus and stomach. Thus, GC-C appears to be an immunohistochemical marker for identifying adenocarcinomas of unknown origin, metastases in patients undergoing staging for GI adenocarcinomas, and intestinal metaplasia, dysplasia, and tumors arising therein in the upper GI tract. PMID- 15754295 TI - p57KIP2 immunohistochemistry in early molar pregnancies: emphasis on its complementary role in the differential diagnosis of hydropic abortuses. AB - Morphologic examination still forms the main diagnostic tool in the differential diagnosis of molar pregnancies. However, the criteria are subjective and show considerable interobserver variability among pathologists. Once a diagnosis of molar pregnancy is made, DNA ploidy studies help to differentiate a triploid partial mole from diploid complete mole (CM). However, with earlier diagnosis and therapeutic evacuation of molar pregnancies, the differentiation of molar pregnancies from early nonmolar placentation is becoming increasingly difficult. The p57(KIP2) gene ( CDKN1C ) is strongly paternally imprinted and expressed from the maternal allele. Because CM lacks a maternal genome, p57(KIP2) immunostaining is correspondingly absent, whereas hydropic abortuses and partial mole show positive staining. We compared the use of p57(KIP2) staining in the differential diagnosis of 68 morphologically challenging cases of early first-trimester hydropic placentas. Diagnosis based on p57(KIP2) staining was compared with the original diagnosis based on morphology and DNA ploidy analysis. Concordant results were obtained in 65 of 68 cases studied. In 2 of 3 cases with a discordant diagnosis, microsatellite DNA genotyping analysis agreed with the results of p57(KIP2) staining, confirming that positive p57(KIP2) staining is a highly sensitive and specific marker for excluding CM in this setting. In addition, p57(KIP2) staining has the advantage of differentiating hydropic abortuses from CMs, a distinction not made by ploidy analysis. p57(KIP2) staining can be used in concert with ploidy studies to refine the diagnosis of early molar pregnancies. PMID- 15754296 TI - p63 overexpression associates with poor prognosis in head and neck squamous cell carcinoma. AB - p63 belongs to a protein family that includes 2 structurally related proteins, p53 and p73. The aim of this study was to investigate the biologic role of p63 in oral tumorigenesis and its possible role as prognostic marker in oral cancer. Ninety-four cases of oral squamous cell carcinoma and 10 cases of normal mucosa were analyzed for p63 expression by immunohistochemistry. Normal oral mucosa showed a basal and parabasal expression of p63. Five (5.3%) cases of oral cancer showed less than 10% of positive tumor cells; in 33 (35.1%) cases the positive tumor cells comprised between 10% and less than 30%, in 36 (38.3%) cases the positive tumor cells comprised between 30% and less than 50%, and in 20 (21.3%) cases the positive tumor cells were more than 50%. There was also a statistically significant correlation between p63 expression and tumor differentiation: p63 expression was amplified in poorly differentiated tumors (P < .05). When analyzed for prognostic significance, patients with perineural infiltration had poorer survival rates than the group with no perineural infiltration (P < .05) and patients with increased p63 expression had poorer survival rates than the group with reduced p63 expression (P < .05). The statistical analysis showed no significant correlation between p63 expression, sex, age, tumor size, staging, recurrence, and metastasis. Cases with diffuse p63 expression were more aggressive and poorly differentiated and related to a poorer prognosis. These data suggest that p63 expression may be useful to identify cases of oral squamous cell carcinoma with more aggressive and invasive phenotype providing novel diagnostic and prognostic information on individual patient survival with oral cancers. PMID- 15754297 TI - The utility of calretinin, inhibin, and WT1 immunohistochemical staining in the differential diagnosis of ovarian tumors. AB - Calretinin has been proposed as a novel marker of ovarian sex cord-stromal tumors (SCST); this study aims to determine whether calretinin can complement or supplant the established utility of inhibin in the differential diagnosis of SCST. WT1 has been shown to be expressed in ovarian serous, but not mucinous neoplasms; its expression in a variety of ovarian tumors is also examined. Formalin-fixed, paraffin-embedded archival tissues from 111 primary ovarian tumors were analyzed with commercially available antibodies using semi-automated immunohistochemistry. Results were graded on a 4-tiered scale with staining of more than 0 but less than 5% of cells considered focal. Of 27 SCST, 56% were calretinin and 56% inhibin positive overall; 90% of granulosa cell tumors, 57% of Sertoli-Leydig cell tumors, 33% of thecomas, and 14% of fibromas were calretinin positive. Inhibin was expressed in 60% of granulosa cell tumors, 71% of Sertoli Leydig cell tumors, 43% of fibromas, and 33% of thecomas. Of 35 surface epithelial tumors (SET), 8% of serous papillary tumors were calretinin positive, whereas 8% of serous papillary tumors and 13% of poorly differentiated carcinomas expressed inhibin. WT1 was expressed in 29% of all endometrioid carcinomas, 10% of borderline mucinous tumors, and no mucinous carcinomas; however, most of the other SETs were positive (77% serous papillary and 88% poorly differentiated carcinomas). Among the SCST, WT1 stained only granulosa cell tumors (75%), though often weakly or variably. Calretinin has only slightly greater sensitivity (76% versus 65%) and equal specificity to inhibin (92%) in the differential staining of granulosa or Sertoli-Leydig cell tumors, that is, nonstromal SCST. Hence, calretinin cannot replace but could complement inhibin as part of an immunohistochemical panel used for diagnostically challenging SCST. Although WT1 should be reliably positive in non-mucinous SET, staining of granulosa cell tumors and lack of expression in a sizable subset of endometrioid carcinomas may confound interpretation. PMID- 15754298 TI - A "floral" variant of nodal marginal zone lymphoma. AB - We describe 6 cases of a specific variant of nodal marginal zone lymphoma with "floral" lymph follicles in patients ranging in age from 18 to 66 years. All 6 patients had lymphadenopathy, either local (n = 5) or systemic (n = 1), and good performance status (0), and none had fever, weight loss, or night sweating. They all underwent excisional biopsy. Histologically, all lesions had a distinctive morphology, with proliferation of medium-sized atypical lymphoid cells in the marginal zone, hyperplastic lymph follicles with enlarged germinal centers, and a thickened mantle zone. In places, folliculolysis was observed. On immunohistochemical staining, the atypical lymphoid cells showed a B-cell phenotype (CD20 +), IgM positivity in 2 of 5 cases, and negativity for CD5, CD10, CD23, CD43, bcl-6, and IgD. Polymerase chain reaction examination for immunoglobulin heavy chain in 5 cases showed monoclonality in all. Five patients did not receive adjuvant chemotherapy and had no recurrences. The patient with systemic lymphadenopathy received chemotherapy and had a complete response without relapse. This variant should be differentiated from the usual nodal marginal zone lymphoma because of its specific clinical and pathological features. PMID- 15754299 TI - Classic follicular dendritic reticulum cell tumor of the lymph node developing in a patient with a previous inflammatory pseudotumor-like proliferation. AB - Inflammatory pseudotumor (IPT) and follicular dendritic reticulum cell tumor (FDRCT) are rare entities of the lymph node characterized by spindle-cell proliferation. We report a case of a 31-year-old woman, who was admitted for biopsy of a lymph node in the left submandibular area. The microscopic examination revealed a proliferation of spindle cells, partially replacing the normal lymph node architecture, suggestive of an IPT. The preserved peripheral portion showed follicular hyperplasia with Castleman-like appearance. Six years later she presented with a new enlargement in the same submandibular area. The nodule was removed, and a diagnosis of a classic FDRCT of the lymph node was made. The present case is remarkable, and clinicopathological data show that IPT like proliferations could be in some case an early presentation of FDRCT. PMID- 15754300 TI - Vascular lesion in a patient of chronic active Epstein-Barr virus infection with hypersensitivity to mosquito bites: vasculitis induced by mosquito bite with the infiltration of nonneoplastic Epstein-Barr virus-positive cells and subsequent development of natural killer/T-cell lymphoma with angiodestruction. AB - This report describes a vasculitis and subsequently developing angiodestructive lymphoma in an 11-year-old Japanese-Filipino girl exhibiting mosquito allergy with the background of chronic active Epstein-Barr virus (EBV) infection. She developed necrotic skin ulcer at the site of mosquito bite, and histopathological examination revealed EBV-positive mononuclear cell infiltration throughout the wall of small-sized muscular artery. These EBV-positive lymphoid cells were oligoclonal in Southern blot analysis for EBV terminal repeats. Effectiveness of steroid therapy also supports the nonneoplastic nature. Approximately 1 year later, she developed progressive large skin ulcer without mosquito bites. Microscopically, the angiocentric or angiodestructive pattern of EBV-positive atypical cells supported the diagnosis of extranodal natural killer/T-cell lymphoma. Southern blot analysis revealed the monoclonal neoplastic nature of EBV positive cells. In contrast to the primary mosquito bite lesion, natural killer/T cell lymphoma cells exhibited the higher expression of EBV latent membrane protein 1 mRNA and the apparent protein expression detected by immunohistochemistry. PMID- 15754301 TI - Adenocarcinoma of the thymus: mucinous subtype. AB - Primary thymic adenocarcinoma, mucinous subtype, is extremely rare with only one case reported to date. We describe herein a case of thymic mucinous adenocarcinoma. A 59-year-old man was identified to have an anterior mediastinal tumor and was diagnosed as mucinous adenocarcinoma. Clinical and radiographic examinations disclosed no evidence of tumor elsewhere. The patient received radiotherapy, but the general condition deteriorated and died 11 months after tumor detection. Thoracic autopsy revealed an anterior mediastinal tumor measuring greater than 10 cm, uncapsulated, and white. The tumor had clear margins and was clearly isolated from the lung. Histologically, the tumor demonstrated papillary, acinar, and cribriform structure and produced abundant extracellular mucin. Immunohistochemically, most tumor cells were positive for cytokeratin 7, were partially positive for CD5, and were negative for TTF-1, Sp A, CDX-2, MUC2, napsin A, and cytokeratin 20. Collectively, the diagnosis of the tumor was primary mucinous adenocarcinoma of the thymus. We propose that the mucinous subtype should be recognized as one of the histopathological entities of thymic adenocarcinoma. PMID- 15754302 TI - A simple method for faster nonbonded force evaluations. AB - Accurate approximations are introduced for the evaluation of inverse interparticle distances such that square root or division operations are not required in the computation of interparticle interactions and forces. These generally applicable approximations are illustrated by incorporation into the protein simulation package TINKER along with several other speed enhancement strategies. With these modifications, implicit solvent Langevin dynamics simulations of proteins are performed factors of 4.6 times faster than the modified open source distributed program. Programming speedups are obtained by extensive vectorization, simplification of the inner loop to avoid IF statements, and by using lookup tables for the distance dependent "dielectric constant" in implicit solvent models. Benchmarks are provided for the all-atom, implicit solvent dynamics of Met-enkephalin, the villin headpiece, the B1 domain of protein-G, and barnase. We also discuss the more general applicability of the approximation methods to explicit solvent simulations and of look-up tables for other implicit solvent models such as the generalized Born models. PMID- 15754304 TI - Agreement between experiment and hybrid DFT calculations for O--H bond dissociation enthalpies in manganese complexes. AB - Information on the accuracy of DFT functionals for redox reactions in transition metal systems is rather limited. To analyze the performance of some popular functionals for redox reactions in manganese systems, calculated O--H bond dissociation enthalpies for Mn-ligands in six different complexes are compared to experimental results. In this benchmark, B3LYP performs well with a mean absolute error of 3.0 kcal/mol. B98 gives similar results to B3LYP (error of 3.8 kcal/mol). B3LYP* gives lower O--H bond strengths than B3LYP and has a mean error of 5.0 kcal/mol. Compared to B98 and B3LYP, B3LYP* has an error trend for the manganese ligands that is more similar to the error for a free water molecule. The nonhybrid functional BLYP consistently and significantly underestimates the O -H bond strengths by approximately 20 kcal/mol. HCTH407 has a rather large mean error of 9.4 kcal/mol and shows no consistent trend. The results support the use of hybrid functionals and the present computational method for large model systems containing manganese. An example is the oxygen evolving complex in photosystem II where hybrid functionals predict the appearance of a Mn(IV)-oxyl radical before the O--O bond formation step. PMID- 15754303 TI - Relative contributions of desolvation, inter- and intramolecular interactions to binding affinity in protein kinase systems. AB - In several previous studies, we performed sensitivity analysis to gauge the relative importance of different atomic partial charges in determining protein ligand binding. In this work, we gain further insights by decomposing these results into three contributions: desolvation, intramolecular interactions, and intermolecular interactions, again based on a Poisson continuum electrostatics model. Three protein kinase-inhibitor systems have been analyzed: CDK2 deschloroflavopiridol, PKA-PKI, and LCK-PP2. Although our results point out the importance of specific intermolecular interactions to the binding affinity, they also reveal the remarkable contributions from the solvent-mediated intramolecular interactions in some cases. Thus, it is necessary to look beyond analyzing protein-ligand interactions to understand protein-ligand recognition or to gain insights into designing ligands and proteins. In analyzing the contributions of the three components to the overall binding free energy, the PKA-PKI system with a much larger ligand was found to behave differently from the other two systems with smaller ligands. In the former case, the intermolecular interactions are very favorable, and together with the favorable solvent-mediated intramolecular interactions, they overcome the large desolvation penalties to give a favorable electrostatics contribution to the overall binding affinity. On the other hand, the other two systems with smaller ligands only present modest intermolecular interactions and they are not or are only barely sufficient to overcome the desolvation penalty even with the aid of the favorable intramolecular contributions. As a result, the binding affinity of these two systems do not or only barely benefit from electrostatics contributions. PMID- 15754305 TI - Empirical force-field assessment: The interplay between backbone torsions and noncovalent term scaling. AB - The kinetic and thermodynamic aspects of the helix-coil transition in polyalanine based peptides have been studied at the ensemble level using a distributed computing network. This study builds on a previous report, which critically assessed the performance of several contemporary force fields in reproducing experimental measurements and elucidated the complex nature of helix-coil systems. Here we consider the effects of modifying backbone torsions and the scaling of noncovalent interactions. Although these elements determine the potential of mean force between atoms separated by three covalent bonds (and thus largely determine the local conformational distributions observed in simulation), we demonstrate that the interplay between these factors is both complex and force field dependent. We quantitatively assess the heliophilicity of several helix stabilizing potentials as well as the changes in heliophilicity resulting from such modifications, which can "make or break" the accuracy of a given force field, and our findings suggests that future force field development may need to better consider effect that vary with peptide length. This report also serves as an example of the utility of distributed computing in analyzing and improving upon contemporary force fields at the level of absolute ensemble equilibrium, the next step in force field development. PMID- 15754306 TI - Enhanced efficacy of conditionally replicating herpes simplex virus (G207) combined with 5-fluorouracil and surgical resection in peritoneal cancer dissemination models. AB - BACKGROUND: The therapeutic efficacy of G207, a replication-competent herpes simplex virus, for malignancies is increased when combined with certain chemotherapies, but the mechanism is unclear and the interaction between G207 and surgical resection has not been extensively studied. The goals of the current study were to examine the performance of combination treatments for peritoneal disseminated cancers and to explore the mechanism of effective combinations. METHODS: Hamsters and SCID and BALB/c mice harboring peritoneal dissemination of gallbladder, gastric or colon cancer cells were treated with G207, 5-fluorouracil (5FU), or surgical resection alone, or G207 combined with 5FU or surgery. Animal survival, antiviral immunity, intratumoral ribonucleotide reductase activity, and viral spread were compared between the groups. RESULTS: The combination of G207 and 5FU prolonged the survival of hamsters bearing peritoneal dissemination of gallbladder cancer compared with the controls, G207 alone and 5FU alone. 5FU did not suppress the production of neutralizing antibodies against G207, but increased ribonucleotide reductase activity and viral spread in subcutaneous gallbladder tumors. The enhanced efficacy of the combination treatment was also observed in immunodeficient mice with disseminated gastric cancer. Although surgical resection did not significantly prolong animal survival or increase the intratumoral activity of ribonucleotide reductase, long-term survivors emerged from groups of animals treated with surgical resection and G207 for gallbladder and colon disseminated cancers. CONCLUSIONS: These results indicate that the increased activity of ribonucleotide reductase in tumors mediated by 5FU and the decreased tumor burden resulting from surgical resection may enhance the therapeutic efficacy of oncolytic herpes virus for peritoneal disseminated cancer. PMID- 15754311 TI - Protonated chiral catalysts: versatile tools for asymmetric synthesis. PMID- 15754307 TI - Poor intercellular transport and absence of enhanced antiproliferative activity after non-viral gene transfer of VP22-P53 or P53-VP22 fusions into p53 null cell lines in vitro or in vivo. AB - BACKGROUND: The herpes simplex virus type 1 (HSV-1) VP22 protein has the property to mediate intercellular trafficking of heterologous proteins fused to its C- or N-terminus. We have previously shown improved delivery and enhanced therapeutic effect in vitro and in vivo with a P27-VP22 fusion protein. In this report, we were interested in studying the spread and biological activity of VP22 fused to the P53 tumor suppressor. METHODS: Expression of the VP22-P53 and P53-VP22 fusion proteins was shown by Western blot and intercellular spreading was monitored by immunofluorescence on transiently transfected cells. In vitro antiproliferative activity of wild-type (wt) P53 and P53-VP22 was assessed by proliferation assays and transactivating ability was studied by a reporter gene test and a gel-shift assay. Antitumor activity was also tested in vivo by intratumoral injections of naked DNA in a model of subcutaneous tumors implanted in nude mice. RESULTS: Our results show that the C-terminal fusion or the N-terminal P53-VP22 fusion proteins are not able to spread as efficiently as VP22. Moreover, we demonstrate that VP22-P53 does not possess any transactivating ability. P53-VP22 has an antiproliferative activity, but this activity is not superior to the one of P53 alone, in vitro or in vivo. CONCLUSIONS: Our study indicates that a gene transfer strategy using VP22 cannot be considered as a universal system to improve the delivery of any protein. PMID- 15754312 TI - Transcriptional regulation of connexin 43 expression by retinoids and carotenoids: similarities and differences. AB - Gap junctions, connexons, are formed by assembly of trans-membrane connexin proteins and have multiple functions including the coordination of cell responses. Most human tumors are deficient in gap junctional communication (GJC) and restoration of GJC by forced expression of connexins reduces indices of neoplasia. Expression of connexin 43 (Cx43), the most widely-expressed connexin family member, is upregulated by cancer-preventive retinoids and carotenoids in normal and preneoplastic cells; an action considered of mechanistic significance. However, the molecular mechanism for upregulated expression is poorly understood. The retinoic acid receptor antagonist Ro 41-5253 was capable of suppressing retinoid-induction Cx43 luciferase reporter construct in F9 cells, but did not suppress reporter activity induced by the non-pro-vitamin A carotenoids astaxanthin or lycopene, indicating that retinoids have separate mechanisms of gene activation than non-pro-vitamin A carotenoids. Neither class of compound required protein synthesis for induction of Cx43 mRNA, nor was the 5.0 h half life of Cx43 mRNA altered, indicating direct transcriptional activation. The responsive region was found within -158 bp and +209 bp of the transcription start site; this contains a Sp1/Sp3 GC-box to which Sp1 and Sp3 were bound, as revealed by electrophoretic mobility shift assays (EMSA), but no retinoic acid response element (RARE). Site directed mutagenesis of this GC-box resulted in increased basal levels of transcription and loss of responsiveness to a synthetic retinoid. In this construct astaxanthin and lycopene produced marginally, but not significantly higher, reporter activity than the control. PMID- 15754313 TI - Localization of the human hedgehog-interacting protein (Hip) in the normal and diseased pancreas. AB - Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a poor prognosis. Previously, it has been shown that Indian hedgehog (Ihh) and its two signaling receptors patched (Ptc) and smoothened (Smo) are involved in the pathogenesis of chronic pancreatitis (CP) and PDAC. In the current study we analyzed the expression, distribution, and function of another component of this signaling pathway, the human hedgehog-interacting protein (Hip), in the normal pancreas, CP and PDAC utilizing real-time quantitative reverse transcription polymerase chain reaction (QRT-PCR), immunohistochemistry, immunofluorescence, Hip siRNA transfection, cell growth assays, and cell cycle analysis. By QRT-PCR, Hip mRNA levels were fifteenfold and fourteenfold increased in CP (n = 22) and PDAC (n = 31) tissues, respectively, compared to normal pancreatic tissues (n=20) and correlated with glioma associated antigen (Gli1) but not Ptc or Protein kinase A (PKA) mRNA levels. Only SU-8686 and BxPC-3 pancreatic cancer cells expressed Hip mRNA, whereas expression was below the level of detection in the other six pancreatic cancer cell lines tested. As shown by immunohistochemistry, Hip was expressed in normal pancreatic tissues mainly in the cytoplasm of islet cells and in smooth muscle cells of blood vessels. In contrast, in CP and PDAC there was a different distribution and staining intensity within the islets. Moreover, Hip immunoreactivity was observed in the tubular complexes, PanIN 1-3 lesions, as well as in pancreatic cancer cells. Incubation of pancreatic cancer cell lines with recombinant Hip revealed a growth inhibitory effect in SU-8686 and Capan-1 pancreatic cancer cells and no effect on cell growth in the other tested cell lines. In addition, silencing of Hip expression using specific siRNA molecules increased the growth of SU-8686 cells. In conclusion, Hip is expressed in the normal pancreas, CP and PDAC tissues. The different pattern of Hip expression and abnormal localization in the diseased pancreas suggest that the enhanced activation of hedgehog signaling in CP and PDAC is-at least in part-due to the aberrant responsiveness and expression of Hip in these diseases. PMID- 15754314 TI - Characterization of the nuclear import of human MutLalpha. AB - DNA mismatch repair (MMR) is essential for the maintenance of replication fidelity. Its major task is to recognize mismatches as well as insertion/deletion loops of newly synthesized DNA strands. Although different players of human MMR have been identified, the regulation of essential steps of MMR is poorly understood. Because MMR is initiated in the nucleus, nuclear import might be a mechanism to regulate MMR. Nuclear targeting is accomplished by conserved signal sequences called nuclear localization signals (NLS), which represent clusters of positively charged amino acids (aa). hMLH1 contains two clusters of positively charged amino acids, which are candidate NLS sequences (aa 469-472 and 496-499), while hPMS2 contains one (aa 574-580). To study the effect of these clusters on nuclear import, NLS mutants of hMLH1 and hPMS2 were generated and expressed in 293T cells. The subcellular localization of the mutant constructs was monitored by confocal laser microscopy. We demonstrated that missense mutations of two signal sequences, one in hMLH1 and one in hPMS2, lead to impaired nuclear import, which was especially prominent for mutants of the hMLH1 residues K471 and R472; and hPMS2 residues K577 and R578. PMID- 15754315 TI - Association of polymorphisms in the cyclin D1 and XPD genes and susceptibility to cancers of the upper aero-digestive tract. AB - DNA repair enzyme genetic polymorphisms have been postulated to increase the risk of certain cancers in the presence of tobacco carcinogen exposures. The XPD protein is an important component of the TFIIH transcription factor complex. XPD genetic polymorphisms resulting in amino acids substitutions may lead to alterations in TFIIH helicase activity, resulting in repair and transcription defects. Cyclin D1 is a key regulatory protein for the transition of cells from the G(1)-S cell cycle phase. The CCND1 G870A polymorphism has been reported to enhance alternate splicing of a stable mRNA variant, which may result in the bypass of the G(1)/S cell cycle checkpoint. In this study, XPD G23591A (Asp312Asn) and A35931C (Lys751Gln) polymorphisms and the CCND1 G870A splice variant frequencies were determined in 273 upper aero-digestive tract cancer cases and 269 controls. The XPD Asp312Asn variant frequency was significantly different among cases and controls and conferred an odds ratio (OR) of 1.3 (95% CI 1.0-1.8). However, individuals with the CCND1 G870A and XPD Lys751Gln variants had higher age adjusted ORs of 3.2 (95% CI 2.2-4.6) and 2.2 (95% CI 1.5-3.2), respectively. Furthermore, a significant gene-gene interaction was observed among cases with at least two variant alleles for both CCND1 and XPD genes [OR 7.09 (95% CI 4.03-12.5)]. Smokers with a combination of at least one variant allele of both CCND1 and XPD genes also had an elevated risk as compared to nonsmokers. This is the first study to suggest an associative interaction between XPD and CCND1 genetic polymorphisms, tobacco exposure, and cancer risk. PMID- 15754316 TI - Experimental and mathematical study of the influence of growth factors on the growth kinetics of adult human articular chondrocytes. AB - This study aimed at determining how kinetic parameters of adult human articular chondrocytes (AHAC) growth are modulated by the growth factor combination TGFbeta1, FGF-2, and PDGF BB (TFP), recently shown to stimulate AHAC proliferation. AHAC, isolated from cartilage biopsies of three individuals, were cultured in medium without (CTR) or with TFP. For growth curves, AHAC were seeded at 1,000 cells/cm(2) and cultured for 12 days, with cell numbers measured fluorimetrically in the same wells every 12 h. For microcolony tests, AHAC were seeded at 2.5 cells/cm(2) and cultured for 6 days, with cell numbers determined for each microcolony by phase contrast microscopy every 8 h. A mathematical model combining delay and logistic equations was developed to capture the growth kinetic parameters and to enable the description of the complete growth process of the cell culture. As compared to CTR medium, the presence of TFP increased the number of cells/well starting from the fifth day of culture, and a four-fold larger cell number was reached at confluency. For single microcolonies, TFP reduced the time for the first cell division by 26.6%, the time for subsequent cell divisions (generation time) by 16.8%, and the percentage of quiescent cells (Q(c)) by 42.5%. The mathematical model fitted well the experimental data of the growth kinetic. Finally, using both microcolony tests and the mathematical model, we determined that prolonged cell expansion induces an enrichment of AHAC with shorter first division time, but not of those with shorter generation time. PMID- 15754317 TI - Functional shape of the skull in vertebrates: which forces determine skull morphology in lower primates and ancestral synapsids? AB - In order to determine the extent to which the shape of the synapsid skull is adapted for resisting the mechanical loads to which it is subjected, block- or simple plate-shaped finite-element models were constructed and loaded with external muscle and bite forces in locations estimated to resemble points of application of these forces. These 2D or 3D finite-element models were iteratively loaded and modified by removing elements that experience only low stresses, and the resulting morphologies of the models were compared with fossil skulls of synapsids and the skulls of extant mammals. The results suggest that the stress flows in these unspecific models are very similar to the arrangement of bone material in real skulls. Morphological differences between taxa depend on a few a priori conditions: length and position of the tooth rows in relation to the braincase, arrangement of muscles, position of the orbits, and position of the nasal opening. Given these initial conditions, finite-element analysis consistently reveals the close similarity between stress flows and real skulls. The major difference between mammal-like reptiles and primates is the size of the braincase. This difference accounts for most of the morphological divergence. The postorbital bar seems to be a constructional element of the skull, rather than a means to protect the eyes. The skull shapes of higher primates are determined mainly by masticatory forces and less by external forces acting on the head. This study demonstrates the utility of finite-element modeling for testing hypotheses regarding relationships between form and function in vertebrate skulls. PMID- 15754318 TI - Adenovirus-mediated inhibition of survivin expression sensitizes human prostate cancer cells to paclitaxel in vitro and in vivo. AB - BACKGROUND: Improvements in the response rates to chemotherapy would represent an important advancement in the care of patients with metastatic prostate cancer. There is accumulating evidence that Survivin, a member of the inhibitor of apoptosis (IAP) family, is associated with both cancer progression and drug resistance. The purpose of this study is to investigate the role of Survivin in paclitaxel-resistance and whether the targeting of Survivin sensitizes prostate cancer cells to paclitaxel. METHODS: Human prostate cell lines PC-3, DU-145, and LNCaP were infected with replication-deficient adenoviruses encoding either wild type Survivin [pAd-S(WT)], to examine Survivin overexpression effects, or a phosphorylation-defective Survivin Thr34 --> Ala dominant negative mutant [pAd S(T34A)], to examine Survivin inactivation effects. The effects of wild-type or mutant Survivin on spontaneous and paclitaxel-induced apoptosis were investigated both in vitro and in vivo. RESULTS: Forced overexpression of wild-type Survivin with pAd-S(WT) increased resistance to paclitaxel in all cell lines, both in vitro and in vivo. Inhibition of Survivin using pAd-S(T34A) resulted in a significant increase in the rate of spontaneous and paclitaxel-induced apoptosis in all cell lines, both in vitro and in vivo. This effect was abolished by co treatment with VAD-CHO (Calbiochem, San Diego, CA), a pan-caspase inhibitor, indicating that Survivin normally mediates resistance to paclitaxel through suppression of caspase-mediated apoptosis. CONCLUSIONS: Survivin mediates paclitaxel-resistance in prostate cancer cells. The inhibition of Survivin sensitizes prostate cancer cells to paclitaxel-induced apoptosis through a caspase-dependant mechanism in vitro and in vivo. PMID- 15754319 TI - pH modulation using CsCl enhances therapeutic effects of vitamin D in LNCaP tumor bearing mice. AB - BACKGROUND: The downstream effects of pH modulation significantly impact the biological fate of chemotherapeutic agents and tumor responsiveness to therapy. We have studied the effects of cesium chloride (CsCl) on pH modulation and subsequent vitamin D treatment in vitamin D receptor (VDR) positive LNCaP tumors and VDR null MDA/LCC6 tumors in vivo. METHODS: Mice bearing LNCaP or MDA/LCC6 tumors were dosed orally with CsCl (150 mg/kg) or vitamin D (1 microg/kg) alone and in combination. Tumor volume and serum PSA (LNCaP only) were measured and intracellular pH was determined, using magnetic resonance spectroscopy (MRS), at tumor and leg muscle sites. Atomic absorption spectroscopy (AAS) was used to quantitate cesium in serum, organs, and tumor tissues. RESULTS: From day 10 onwards, statistically significant (P<0.01) differences were observed in all LNCaP treated groups as compared with control. CsCl co-administered with vitamin D, caused an apparent sensitization of efficacy in this tumor model. There were no correlating differences in serum PSA. Elevation of pH was statistically significant for all three treatment groups as compared with control. The pH measured in leg muscle was not influenced by CsCl treatment. Inhibition of tumor growth was not apparent in VDR null MDA/LCC6 tumors although intracellular tumor pH was shifted. Cesium was rapidly absorbed into serum and present in LNCaP tumors, prostates and other tissues after 1 hr, remaining for up to 24 hr. CONCLUSIONS: The data presented in this manuscript is the first report of chemosensitization by in vivo pH modulation using CsCl in mice bearing prostate or any other tumor xenograft. PMID- 15754320 TI - Ultrastructural characterization of an artificial basement membrane produced by cultured keratinocytes. AB - A recent study in our laboratories on the growth of keratinocytes at the culture medium/air interface has led to the identification of a novel thin sheet-like matrix that supports adherent cells. This novel matrix consists of components secreted by keratinocytes, including type IV collagen, and laminins 1 and 5, that self-assembled to a membrane structure. In the present study, a detailed ultrastructural characterization of this membrane was done with high-resolution electron microscopy after negative staining. The basic organization of the membrane was found to be a dense network of 8- to 10-nm-wide irregular rod-like elements. High-resolution examination and immunolabeling showed that type IV collagen filaments form the core of these elements, and other components including heparan sulfate proteoglycan in the form of 4.5- to 5-nm-wide ribbon like "double tracks" are aggregated around it. These detailed features of the membrane strikingly resembled those of the basement membrane in vivo. These ultrastructural similarities indicate that the membrane may also have basement membrane-like functional properties, and suggest that it should be considered for testing in future medical applications. PMID- 15754321 TI - p75NTR-mediated signaling promotes the survival of myoblasts and influences muscle strength. AB - During muscle development, the p75(NTR) is expressed transiently on myoblasts. The temporal expression pattern of the receptor raises the possibility that the receptor is influencing muscle development. To test this hypothesis, p75(NTR) deficient mutant mice were tested for muscle strength by using a standard wire gripe strength test and were found to have significantly decreased strength relative to that of normal mice. When normal mybolasts were examined in vivo for expression of NGF receptors, p75(NTR) was detected on myoblasts but the high affinity NGF receptor, trk A, was not co-expressed with p75(NTR). In vitro, proliferating C2C12 and primary myoblasts co-expressed the p75(NTR) and MyoD, but immunofluorescent analysis of primary myoblasts and RT-PCR analysis of C2C12 mRNA revealed that myoblasts were devoid of trk A. In contrast to the cell death functions that characterize the p75(NTR) in neurons, p75(NTR)-positive primary and C2C12 myoblasts did not differentiate or undergo apoptosis in response to neurotrophins. Rather, myoblasts survived and even proliferated when grown at subconfluent densities in the presence of the neurotrophins. Furthermore, when myoblasts treated with NGF were lysed and immunoprecipitated with antibodies against phosphorylated I-kappaB and AKT, the cells contained increased levels of both phospho-proteins, both of which promote cell survival. By contrast, neurotrophin-treated myoblasts did not induce phosphorylation of Map Kinase p42/44 or p38, indicating the survival was not mediated by the trk A receptor. Taken together, the data indicate that the p75(NTR) mediates survival of myoblasts prior to differentiation and that the activity of this receptor during myogenesis is important for developing muscle. PMID- 15754322 TI - The non-receptor tyrosine kinase Syk regulates lamellipodium formation and site directed migration of human leukocytes. AB - The tyrosine kinase Syk is associated with CD18, the beta-subunit of the leukocyte adhesion molecules of the beta(2) integrin family (CD11/CD18), and becomes activated upon beta(2) integrin-mediated adhesion. In this study, we elucidated the role of Syk in polarization and site-directed migration of neutrophil-like differentiated HL-60 cells and monocytic THP-1 cells. By means of confocal microscopy, we detected a homogenous distribution of Syk in unstimulated cells in suspension. The stimulation of HL-60 cells by formyl-methionyl-leucyl phenylalanine (fMLP, 100 nM) or the activation of THP-1 cells by monocyte chemoattractant protein-1 (10 ng/ml) induced beta(2) integrin-mediated cell adhesion and polarization on immobilized fibrinogen which was associated with an enrichment of Syk at the lamellipodium forming site. This effect was abolished by function blocking anti-CD18 antibody or by treatment of the cells with the Syk inhibitor piceatannol (30 microM) suggesting that the redistribution of Syk required both, beta(2) integrin-mediated adhesion and Syk activation. Moreover, the inhibition of Syk by piceatannol or the downregulation of Syk by antisense technique resulted in an excessive formation of lamellipodia indicating that Syk may act as a negative regulator that limits lamellipodium formation. The analysis of chemotaxis revealed that the inhibition of Syk impaired the ability of the cells to follow a chemotactic gradient whereas random migration was intact. Taken together, our data suggest a novel role for Syk in the maintenance of a bipolar phenotype by regulating lamellipodium formation, which is a critical prerequisite for site-directed migration of leukocytes. PMID- 15754323 TI - Finite element analysis in vertebrate biomechanics. AB - This special issue of The Anatomical Record presents a series of papers that apply the method of finite element analysis (FEA) to questions in vertebrate biomechanics. These papers are salient examples of the use of FEA to test hypotheses regarding structure-function relationships in complexly shaped biological objects such as skulls and in areas of the skeleton that are otherwise impervious to study. FEA is also a powerful tool for studying patterns of stress and strain in fossil animals and artificial constructs hypothesized to represent ancestral conditions. FEA has been used deductively, to study patterns of growth and development, and to investigate whether skull shapes can be created from amorphous blocks using an iterative approach of loading and removing elements. Several of the papers address methodological issues, such as the relative importance of loading conditions and material properties for generating an accurate model and the validation of models using in vivo strain data. Continuing improvements in model building techniques will make possible increased application of FEA to study the functional effects of variation in morphology, whether through ontogenetic or phylogenetic transformations. PMID- 15754324 TI - Indirect regulation of the intestinal H+-coupled amino acid transporter hPAT1 (SLC36A1). AB - A H(+)-coupled amino acid transporter has been characterised functionally at the brush border membrane of the human intestinal cell line Caco-2. This carrier, hPAT1 (human Proton-coupled Amino acid Transporter 1) or SLC36A1, has been identified recently at the molecular level and hPAT1 protein is localised to the brush border membrane of human small intestine. hPAT1 transports both amino acids (e.g., beta-alanine) and therapeutic agents (e.g., D-cycloserine). In human Caco 2 cells, hPAT1 function (H(+)/amino acid symport) is associated with a decrease in intracellular pH (pH(i)), which selectively activates the Na(+)/H(+) exchanger NHE3, and thus maintains pH(i) and the driving force for hPAT1 function (the H(+) electrochemical gradient). This study provides the first evidence for regulation of hPAT1 function. Activation of the cAMP/protein kinase A pathway in Caco-2 cell monolayers either using pharmacological tools (forskolin, 8-br-cAMP, [(11,22,28)Ala]VIP) or physiological activators (the neuropeptides VIP and PACAP) inhibited hPAT1 function (beta-alanine uptake) at the apical membrane. Under conditions where NHE3 is inactive (the absence of Na(+), apical pH 5.5, the presence of the NHE3 inhibitor S1611) no regulation of beta-alanine uptake is observed. Forskolin and VIP inhibit pH(i) recovery (NHE3 function) from beta alanine-induced intracellular acidification. Immunocytochemistry localises NHERF1 (NHE3 regulatory factor 1) to the apical portion of Caco-2 cells where it will interact with NHE3 and allow PKA-mediated phosphorylation of NHE3. In conclusion, we have shown that amino acid uptake via hPAT1 is inhibited by activators of the cAMP pathway indirectly through inhibition of NHE3 activity. PMID- 15754325 TI - Exposure to 50 Hz electromagnetic radiation promote early maturation and differentiation in newborn rat cerebellar granule neurons. AB - The wish of this work is the study of the effect of electromagnetic (EMF) radiations at a frequency of 50 Hz on the development of cerebellar granule neurons (CGN). Granule neurons, prepared from newborn rat cerebellum (8 days after birth), were cultured after plate-seeding in the presence of EMF radiations, with the plan of characterizing their cellular and molecular biochemistry, after exposure to the electromagnetic stimulus. Five days challenge to EMF radiations showed, by the cytotoxic glutamate (Glu) pulse test, a 30% decrease of cells survival, while only 5% of mortality was reported for unexposed sample. Moreover, blocking the glutamate receptor (GluR) with the Glu competitor MK-801, no toxicity effect after CGN challenge to EMF radiations and Glu was detected. By patch-clamp recording technique, the Kainate-induced currents from 6 days old exposed CGN exhibited a significant increase with respect to control cells. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses show that EMF exposure of rats CGN, induces a change in both GluRs proteins and mRNAs expression with respect to control. In addition, the use of monoclonal antibody raised against neurofilament protein (NF-200) reveals an increase in NF-200 synthesis in the exposed CGN. All these results indicate that exposure to non-ionizing radiations contribute to a premature expression of GluRs reducing the life span of CGN, leading to a more rapid cell maturation. PMID- 15754326 TI - Tissue inhibitor of metalloproteinase 1 is an independent predictor of prognosis in patients with nonsmall cell lung carcinoma who undergo resection with curative intent. AB - BACKGROUND: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play a role in the processes of extracellular matrix degradation. Changes in their expression levels have been observed in various tumor types, including lung carcinoma. However, their clinical significance and their prognostic importance in the progression of nonsmall cell lung carcinoma (NSCLC) remain to be specified. In this study, mRNA expression levels of MMP-1, MMP-9, TIMP-1, and TIMP-2 were evaluated in patients with resected NSCLC, and their associations with disease progression and prognosis were determined. METHODS: Between June 1996 and December 1999, 116 patients underwent resection for NSCLC. Expression levels of MMPs and TIMPs were evaluated using Northern blot analysis in these NSCLC tissue samples and in 39 matched samples of normal lung tissue. RESULTS: MMP-1, MMP-9, and TIMP-1 expression levels were increased in tumor samples compared with matched, corresponding normal tissues. In contrast, TIMP-2 expression was decreased in tumor samples. MMP-1 tumor expression was correlated significantly with the evolution of lymph node status and tumor-lymph node-metastasis (TNM) stage. In contrast, MMP-9 tumor expression was correlated significantly with increased T stage. TIMP-1 overexpression was an independent predictor of worse survival in patients with NSCLC that was not associated with other prognosis factors, such as TNM stage. CONCLUSIONS: The overexpression of TIMP-1 was an independent prognostic marker in patients with NSCLC, and evaluating TIMP-1 may be important for identifying patients who are at greater risk of disease recurrence. PMID- 15754327 TI - Bile acids induce MUC2 overexpression in human colon carcinoma cells. AB - BACKGROUND: Mucin alterations are a common feature of colonic neoplasia, and alterations in MUC2 mucin have been associated with tumor progression in the colon. Bile acids have been linked to colorectal carcinogenesis and mucin secretion, but their effects on mucin gene expression in human colon carcinoma cells is unknown METHODS: Human colon carcinoma cells were treated chenodeoxycholate > ursodeoxycholate). Treatment with the putative chemopreventive agent curcumin, which decreased AP-1 activity, also decreased MUC2 transcription. Cotransfection with a dominant negative AP-1 vector decreased MUC2 transcription, confirming the significance of AP-1 in MUC2 induction by deoxycholate. Calphostin C, a specific inhibitor of protein kinase C (PKC), greatly decreased bile acid-induced MUC2 transcription and AP-1 activity, whereas inhibitors of MAP kinase had no effect. CONCLUSIONS: Bile acids induced mucin expression in human colon carcinoma cells by increasing MUC2 transcription through a process involving MAP kinase-independent, PKC dependent activation of AP-1. PMID- 15754329 TI - The role of nitric oxide synthases and nitrotyrosine in retinoblastoma. AB - BACKGROUND: To investigate the potential involvement of the nitric oxide (NO) pathway in retinoblastoma, the authors correlated immunoreactivity for endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), and nitrotyrosine (NT) with the degree of tumor invasiveness in retinoblastoma. METHODS: eNOS, iNOS, and NT reactivity was evaluated by immunohistochemistry in 34 archival retinoblastoma specimens and in a human Y79 retinoblastoma cell line. The tumors were divided into 2 groups: Group A tumors (n = 17 tumors) with no invasion and Group B tumors (n = 17 tumors) with invasion of the choroid, optic nerve, and/or orbit. The expression levels of eNOS, iNOS, and NT were correlated with invasiveness of the tumors. RESULTS: In Group A tumors (n = 17 tumors) without invasion, eNOS was positive in 17 of 17 tumors (100%), iNOS was positive in 14 of 17 tumors (82%), and NT was positive in 17 of 17 tumors (100%). In Group B tumors (n = 17 tumors) with invasion, eNOS was positive in 17 of 17 tumors (100%), iNOS was positive in 16 of 17 tumors (94%), and NT was positive in 17 of 17 tumors (100%). The invasive cohort showed significantly higher expression of iNOS (P < 0.0001) and NT (P < 0.020) compared with the noninvasive cohort. Y79 cells also expressed eNOS, iNOS, and NT; and nonneoplastic retina was positive for eNOS, iNOS, and NT. CONCLUSIONS: Taken together, the results suggested that retinoblastomas can produce NO. The roles of NO in the biology of retinoblastoma and in the prognosis for patients with retinoblastoma remain to be established. PMID- 15754328 TI - SarCNU-induced G2/M arrest in hepatoma cells is mediated by a p53-independent phosphorylation of cdc-2 at Tyr15. AB - Hepatocellular carcinoma (HCC) is a major health problem in the Asia-Pacific region, with high incidence and mortality rate. There is currently no effective treatment for inoperable cases that represent the vast majority of patients. In the present study, we report that in vitro treatment of primary hepatoma, HepG2 (wild-type p53), PLC/PRF/5 (p53-mutant), and Hep3B (p53-deleted) cells with 2 chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU) resulted in upregulation of p53, p21(Cip1/Waf1), phosphorylated cdc-2 at Tyr15 in wild-type p53 cells and phosphorylation of cdc-2 at Tyr15 in p53-mutant or p53-deleted hepatoma cells. This was accompanied by the reduction in cdc-2 kinase activity and G(2)/M cell cycle arrest. These findings indicate that SarCNU-induced G(2)/M growth arrest in hepatoma cells by a p53-independent phosphorylation of cdc-2. Our data suggest the potential use of SarCNU in treatment of HCC. PMID- 15754330 TI - No effect of hydroxyapatite particles in phagocytosable sizes on implant fixation: an experimental study in dogs. AB - The influence of wear debris on bone healing around orthopedic implants is debated. Hydroxyapatite (HA) particles and polyethylene (PE) particles have been shown to have a negative effect on osteoblast cultures in vitro. The present study investigated the in vivo effects of HA and PE particles on the mechanical fixation and gap healing around experimental HA implants. Nonloaded implants (n = 30) were inserted bilaterally into the proximal tibia of 15 dogs with a 2-mm gap to the bone. The peri-implant gap was either (1) empty (n = 6) or filled with (2) hyaluronic acid (n = 8), (3) hyaluronic acid and HA particles (n = 8), or (4) hyaluronic acid and PE particles (n = 8). After 4 weeks, the animals were killed. The implant interface was evaluated by pushout testing until failure and by histomorphometry. Both HA and PE particles were found to be phagocytosed by macrophage-like cells in the interfacial tissue. HA particles were also integrated in newly formed bone. We found no negative effect of the particulate material on mechanical fixation of the implants or on bone formation around the implants. PMID- 15754331 TI - Development and resolution of experimental colitis in mice with targeted deletion of dipeptidyl peptidase IV. AB - Glucagon-like peptide-2 (GLP-2) is a potent intestinotrophic growth factor that enhances repair of damaged intestinal tissue. However, its bioactivity is limited by dipeptidyl peptidase IV (DPIV)-mediated degradation. We hypothesized that DPIV(-/-) mice would display an increased resistance to, and an enhanced recovery from, dextran sulfate sodium (DSS)-induced colitis compared to DPIV(+/+) mice. DPIV(+/+) and DPIV(-/-) mice consumed 2% DSS for 6 days, followed by a 15 day recovery period. Mice were killed at days 0, 3, 6, 9, 14, and 21 (n = 6-8) and the small intestine and colon removed for histological assessment of villus height, crypt depth, and crypt area. The epithelial cell proliferative labeling index was determined by proliferating cell nuclear antigen (PCNA) immunostaining. Small intestine, colon, and total body weight did not differ between DPIV(+/+) and DPIV(-/-) mice. Distal colon crypt depth did not differ significantly between DPIV(+/+) and DPIV(-/-) mice during the development of DSS-colitis or during the recovery phase. Similarly no significant effects were apparent on distal colon crypt area or PCNA labeling index between DPIV(+/+) and DPIV(-/-) during the development of and recovery from DSS-colitis. However, DPIV(-/-) mice still possessed significant levels of plasma DPIV-like activity. We conclude that loss of DPIV activity does not increase resistance to experimental colitis and hypothesize that other DPIV family members may also be involved in the cleavage of GLP-2. PMID- 15754332 TI - Improving delivery of antineoplastic agents with anti-vascular endothelial growth factor therapy. AB - It is believed that impairments in delivery of antineoplastic agents to solid tumors result from abnormalities of the tumor microenvironment. Vascular endothelial growth factor (VEGF), the prototypical angiogenic molecule, is one of the main factors responsible for the development and maintenance of the aberrant tumor vascular network, which is characterized by chaotic, leaky blood vessels with high interstitial fluid pressure and inefficient blood flow. The authors proposed that anti-VEGF therapy would reduce the elevated interstitial fluid pressure in tumors, thereby improving blood flow and potentially improving delivery of cytotoxic agents to tumor cells. For the current report, the authors reviewed characteristics of the abnormal tumor vasculature created under the influence of VEGF, the resulting tumor microenvironment, how the tumor microenvironment may impede delivery of antineoplastic agents, and how the combination of anti-VEGF and cytotoxic therapy may maximize the efficacy of antineoplastic treatment regimens. PMID- 15754333 TI - Therapy-related changes in body size in Hispanic children with acute lymphoblastic leukemia. AB - BACKGROUND: The objective of this study was to examine changes over time in body mass index (BMI) from diagnosis through chemotherapy for pediatric patients with B-precursor acute lymphoblastic leukemia (ALL). METHODS: The study cohort consisted of 141 white Hispanic pediatric patients who were diagnosed with ALL and were treated at 2 South Texas pediatric oncology centers between 1993 and 2002. Changes in age-standardized and gender-standardized BMI scores were assessed. RESULTS: The study cohort exhibited a steady increase in age-adjusted and gender-adjusted BMI scores for the first 12 months of therapy, a modest increase in BMI scores during the 18-23 month and 24-29 month periods, followed by a slight decrease in BMI scores at 30 months (end of therapy). A repeated measures analysis indicated significant effects for time (P = 0.019) and time by baseline BMI category interaction (P = 0.0001) but no significant interaction effect between time and gender (P = 0.65). CONCLUSIONS: Although it is known that leukemia therapy is associated with prevalent obesity in survivorship, its pattern of development during therapy has not been elucidated. In the current cohort of Hispanic children with ALL, BMI scores were elevated at diagnosis (mean +/- standard deviation standardized BMI Z score, 0.33 +/- 1.4), then increased, and remained elevated for the entire duration of chemotherapy. Patients who were classified as normal weight exhibited an increase in BMI over time; patients who were classified as overweight at diagnosis exhibited BMI patterns that were relatively stable; and patients who were classified as obese exhibited a very slight decline over time. These findings suggest that the risk for chemotherapy related weight gain applies predominantly to children who begin ALL therapy within a normal weight range. PMID- 15754334 TI - Mast cell exocytosis can be triggered by ammonium chloride with just a cytosolic alkalinization and no calcium increase. AB - A human mast cell line (HMC-1) has been used to study the effect of cytosolic alkaline pH in exocytosis. Compound 48/80, concanavalin A, and thapsigargin do not induce histamine release in HMC-1 cells. Although thapsigargin does not activate histamine release, it does show a large increase in cytosolic Ca(2+), and no change in cytosolic pH. However, when HMC-1 cells were activated with ionomycin, a significant histamine release takes place, and this effect is higher in the presence of thapsigargin. Both drugs show an additive effect on cytosolic Ca(2+) levels. Ammonium chloride (NH(4)Cl) does activate cytosolic alkalinization and histamine release, with no increase in cytosolic Ca(2+). NH(4)Cl does block the release of internal Ca(2+) by thapsigargin, not by ionomycin, and decreases Ca(2+) influx stimulated by these drugs. Under conditions in which the alkalinization induced by NH(4)Cl is blocked by acidification with sodium propionate, histamine release is inhibited. The release of histamine is also observed when NH(4)Cl is added after propionate addition, regardless of the final pH value attained. Our results show that a shift in pH alkaline values, even with final pH below 7.2 is enough to activate histamine release. A shift to less acidic values is a sufficient signal to activate the cells. PMID- 15754335 TI - The IVADo regimen--a pilot study with ifosfamide, vincristine, actinomycin D, and doxorubicin in children with metastatic soft tissue sarcoma: a pilot study of behalf of the European pediatric Soft tissue sarcoma Study Group. AB - BACKGROUND: The role of doxorubicin (Doxo) as part of multidrug regimens used to treat children with soft tissue sarcoma (STS) is controversial. To evaluate the feasibility of combining Doxo with the well established ifosfamide, vincristine, and actinomycin D (IVA) regimen, the Italian STS Committee performed a pilot study on a series of children with metastatic STS. METHODS: Between July 2002 and February 2004, 29 evaluable patients were enrolled in this study; 19 patients had rhabdomyosarcoma, 5 patients had peripheral neuroectodermal tumor, and 5 patients had other types of STS. The IVA-Doxo (IVADo) regimen included ifosfamide 3 g/m(2) on Days 1 and 2, vincristine 1.5 mg/m(2) on Day 1, actinomycin D 1.5 mg/m(2) on Day 1, and Doxo 30 mg/m(2) on Days 1 and 2. Three courses of IVADo were to be administered in the initial part of treatment and analyzed for toxicity and tumor response. RESULTS: Overall, 92 cycles were delivered. Major regimen-related toxicity was myelosuppression, with Grade 4 neutropenia in 67% of cycles and fever and neutropenia in 37% of cycles. Nonhematologic toxicity included Grade 3 4 mucositis (6.5% of cycles), constipation (9.7%), and peripheral neuropathy (6.5%). Other manifestations of major toxicity were venoocclusive disease and seizures, which occurred in one patient each. All but 1 patient with a malignant schwannoma showed some degree of tumor volume reduction; however, considering only complete and partial remissions, the response rate was 76% (+/- 7.9%). CONCLUSIONS: The intensive IVADo regimen was effective against pediatric STS with acceptable toxicity. This combination will be investigated in high-risk patients with rhabdomyosarcoma in a randomized trial launched by the European pediatric Soft tissue sarcoma Study Group. PMID- 15754336 TI - Pregnancy-associated plasma protein-A proteolytic activity in rat vertebral cell cultures: stimulation by dexamethasone--a potential mechanism for glucocorticoid regulation of osteoprogenitor proliferation and differentiation. AB - Glucocorticoids (GCs) at physiological concentrations stimulate osteoprogenitor proliferation and differentiation in rat bone cell populations, and this is mediated in part by an increased response to insulin-like growth factors (IGFs). Since IGF binding proteins (IGFBPs) modulate IGF actions, we evaluated whether the increased IGF responsiveness might be associated with decreased inhibitory IGFBP-4 peptide levels. Rat vertebral cells were cultured for up to 20 days with or without dexamethasone (Dex). Cell layer proteins were extracted at day 6, 8, 14, and 20, conditioned media (CM) collected at day 8, 14, and 20, and total RNA isolated at day 14 and 20 of culture. Western blotting showed that cell layer IGFBP-4 levels were lower, while IGFBP-4 protease activity in CM was higher, in Dex-treated cultures. Addition of pregnancy-associated plasma protein-A (PAPP-A) antibody to CM abrogated IGFBP-4 proteolysis. PAPP-A mRNA levels were the same in control and Dex-treated cultures as evaluated by RT-PCR. Our data demonstrate that activity of the IGFBP-4 protease, PAPP-A, in rat bone cell cultures is increased by Dex via post-transcriptional mechanisms. Since IGFBP-4 mRNA levels in Dex-treated cultures were the same as in controls at day 8, slightly lower than in controls at day 14, and higher than in controls at day 20 as shown previously, the decreased IGFBP-4 peptide levels in Dex-treated cultures likely result from increased IGFBP-4 proteolysis by the elevated PAPP-A enzymatic activity. Our findings underscore a novel mechanism whereby GCs increase IGF responses in rat bone cells via PAPP-A-induced IGFBP-4 proteolysis. PMID- 15754337 TI - Anti-adipogenic effect of PDGF is reversed by PKC inhibition. AB - Healthy adipose tissue function depends on adipogenesis. The capacity to form new adipocytes prevents the emergence of insulin-resistant hypertrophied adipocytes, as well as the deleterious lipid deposition in muscle, liver, and pancreas. It is therefore important to understand how adipogenesis is modulated. Platelet-derived growth factor (PDGF) is anti-adipogenic, but the stage of differentiation that it targets, and the signaling pathways that it triggers, are not defined. We have studied the inhibitory effect of PDGF on murine 3T3-L1 preadipocyte and human preadipocyte differentiation. There was a significant attenuation in the protein expression of the adipogenic transcription factors, PPARgamma and C/EBPalpha, as well as in the levels of later differentiation markers, including adiponectin, aP2, and fatty acid synthase. PDGF treatment resulted in the persistence of PDGF receptor and PKCalpha expression, in contrast to the expected downregulation of both proteins that occurs during differentiation. Inactivation of conventional PKC isoforms, by bisindolylmaleimide I or PKC pseudosubstrate M20-28, partially reversed the inhibition of 3T3-L1 and human preadipocyte differentiation by PDGF, as assessed by fatty acid synthase expression and morphological appearance. PMID- 15754338 TI - Effect of adenosine triphosphate in renal ischemic injury: involvement of NF kappaB. AB - Renal ischemic/reperfusion injury in vivo results in a significant increase of acute renal failure (ARF) and death. Nevertheless, there are many limitations in using in vivo models of renal ischemic injury to elucidate the detailed mechanisms of renal injury. Adenosine triphosphate (ATP), an extracellular signal, has been shown to be an important factor in regulation of epithelial cell function. Thus, the present study was performed to establish in vitro ischemic model using primary cultured rabbit renal proximal tubule cells (PTCs) and to examine the effect of ATP in this model. We established an in vitro model of ischemic injury, causing severe depletion of intracellular ATP by using the combination of a mitochondrial respiration inhibitor (antimycin A), non metabolizable glucose analog (2-deoxyglucose), and calcium ionophore (A23187) in PTCs. Indeed, this ischemic injury significantly increased LDH release, a marker of structural damage, and ATP blocked ischemic injury-induced LDH release. 2 Methylthio-ATP and ATP-gamma-S (P2Y purinoceptor agonists) also blocked ischemic injury-induced LDH release, whereas AMP-CPP (P2X purinoceptor agonist) did not block it. In experiments to examine the relationship between ischemic injury and NF-kappaB activation, ischemic injury increased NF-kappaB translocation, DNA binding activity, and CAT activity. On the other hand, ATP, ATP-gamma-S, or 2 methylthio-ATP protected ischemic injury-induced NF-kappaB activation. These results suggest that the protective effect of ATP on ischemic injury is, in part, related to inhibition of NF-kappaB activation via P2Y receptor in PTCs. PMID- 15754339 TI - Functional analysis of TMLH variants and definition of domains required for catalytic activity and mitochondrial targeting. AB - epsilon-N-Trimethyllysine hydroxylase (TMLH) (EC 1.14.11.8) is a non-heme-ferrous iron hydroxylase, Fe(++) and 2-oxoglutarate (2OG) dependent, catalyzing the first of four enzymatic reactions of the highly conserved carnitine biosynthetic pathway. Otherwise from all the other enzymes of carnitine biosynthesis, TMLH was found to be associated to the mitochondrial fraction. We here report molecular cloning of two alternative spliced forms of TMLH, which appear ubiquitously expressed in human adult and fetal tissues. The deduced proteins are designated TMLH-a and TMLH-b, and contain 421 and 399 amino acids, respectively. They share the first N-terminal 332 amino acids, including a mitochondrial targeting signal, but diverge at the C-terminal end. TMLH-a and TMLH-b exogenous expression in COS 1 cells shows that the first 15 amino acids are necessary and sufficient for mitochondrial import. Furthermore, comparative evolutionary analysis of the C terminal portion of TMLH-a identifies a conserved domain characterized by a key triad of residues, His242-Glu244-His389 predicted to bind 2OG end. This sequence is conserved in the TMLH enzyme from all species but is partially substituted by a unique sequence in the TMLH-b variant. Indeed, TMLH-b is not functional by itself as well as a TMLH-H389L mutant produced by site directed mutagenesis. As great interest, we found that TMLH-b and TMLH-H389L, individually co-expressed with TMLH-a in COS-1 cells, negatively affect TMLH activity. Therefore, our studies on the TMLH alternative form provide relevant novel information, first that the C-terminal region of TMLH contains the main determinants for its enzymatic activity including a key H389 residue, and second that TMLH-b could act as a crucial physiological negative regulator of TMLH. PMID- 15754340 TI - Electromagnetic fields at mobile phone frequency induce apoptosis and inactivation of the multi-chaperone complex in human epidermoid cancer cells. AB - The exposure to non-thermal microwave electromagnetic field (MW-EMF) at 1.95 MHz, a frequency used in mobile communication, affects the refolding kinetics of eukaryotic proteins (Mancinelli et al., 2004). On these basis we have evaluated the in vivo effect of MW-EMF in human epidermoid cancer KB cells. We have found that MW-EMF induces time-dependent apoptosis (45% after 3 h) that is paralleled by an about 2.5-fold decrease of the expression of ras and Raf-1 and of the activity of ras and Erk-1/2. Although also the expression of Akt was reduced its activity was unchanged likely as a consequence of the increased expression of its upstream activator PI3K. In the same experimental conditions an about 2.5-fold increase of the ubiquitination of ras and Raf-1 was also found and the addition for 12 h of proteasome inhibitor lactacystin at 10 microM caused an accumulation of the ubiquitinated isoforms of ras and Raf-1 and counteracted the effects of MW EMF on ras and Raf-1 expression suggesting an increased proteasome-dependent degradation induced by MW-EMF. The exposure of KB cells to MW-EMF induced a differential activation of stress-dependent pathway with an increase of JNK-1 activity and HSP70 and 27 expression and with a reduction of p38 kinase activity and HSP90 expression. The overexpression of HSP90 induced by transfection of KB cells with a plasmid encoding for the factor completely antagonized the apoptosis and the inactivation of the ras --> Erk-dependent survival signal induced by MW EMF. Conversely, the inhibition of Erk activity induced by 12 h exposure to 10 mM Mek-1 inhibitor U0126 antagonized the effects induced by HSP90 transfection on apoptosis caused by MW-EMF. In conclusion, these results demonstrate for the first time that MW-EMF induces apoptosis through the inactivation of the ras --> Erk survival signaling due to enhanced degradation of ras and Raf-1 determined by decreased expression of HSP90 and the consequent increase of proteasome dependent degradation. PMID- 15754341 TI - Influence of oxygen on the proliferation and metabolism of adipose derived adult stem cells. AB - Articular cartilage is an avascular connective tissue that exhibits little intrinsic capacity for repair. Articular cartilage exists in a reduced oxygen ( approximately 5%) environment in vivo; therefore, oxygen tension may be an important factor that regulates the metabolism of chondrocyte progenitors. A number of recent studies have developed tissue engineering approaches for promoting cartilage repair using undifferentiated progenitor cells seeded on biomaterial scaffolds, but little is known about how oxygen might influence these engineered tissues. Human adipose-derived adult stem (hADAS) cells isolated from the stroma of subcutaneous fat were suspended in alginate beads and cultured in control or chondrogenic media in either low oxygen (5%) or atmospheric oxygen tension (20%) for up to 14 days. Under chondrogenic conditions, low oxygen tension significantly inhibited the proliferation of hADAS cells, but induced a two-fold increase in the rate of protein synthesis and a three-fold increase in total collagen synthesis. Low oxygen tension also increased glycosaminoglycan synthesis at certain timepoints. Immunohistochemical analysis showed significant production of cartilage-associated matrix molecules, including collagen type II and chondroitin-4-sulfate. These findings suggest oxygen tension may play an important role in regulating the proliferation and metabolism of hADAS cells as they undergo chondrogenesis, and the exogenous control of oxygen tension may provide a means of increasing the overall accumulation of matrix macromolecules in tissue-engineered cartilage. PMID- 15754343 TI - Effects of a laminin peptide (YIGSR) immobilized on crab-tendon chitosan tubes on nerve regeneration. AB - Thiolated and nonthiolated hydroxyapatite-coated crab-tendon chitosan (t chitosan/HAp-SH and t-chitosan/HAp, respectively) tubes, both alone and conjugated with CDPGYIGSR (YIGSR) peptide, were compared, in order to determine their biocompatibility and efficacy as nerve conduits. YIGSR peptide was adsorbed on the t-chitosan/HAp (HAp) tubes, and covalently bound on the t-chitosan/HAp-SH (HAp-SH) tubes (Y/HAp and Y/HAp-SH tubes, respectively). HAp, HAp-SH, Y/HAp, or Y/HAp-SH tubes measuring 15 mm were bridge grafted into the sciatic nerve of SD rats. Grafting of 15-mm-long Type I atelocollagen tubes and isografting of sciatic nerves were also carried out (N = 6 in each group). After 12 weeks, evoked muscle action potentials were recorded to calculate the terminal latency quotient. Histological observation and analysis of myelinated axons were also carried out. Nerve-tissue regeneration did not occur directly on the tubes' surfaces in the YIGSR peptide-unconjugated groups. Transplantation of YIGSR conjugated tubes, however, gave rise to regenerated nerve tissue attached to thin layers of epineurium-like structure formed on the inner-tube surface. Histological and electrophysiological analyses suggested that although thiolation retards nerve-tissue regeneration, adsorbed YIGSR, and, to a lesser extent, peptide that had been covalently bound onto the tube surfaces, enhance nerve regeneration, promoting sprouting from the proximal nerve stump and bridging of regenerated axons throughout the tube. PMID- 15754342 TI - Antagonists of growth hormone releasing hormone (GHRH) and of bombesin/gastrin releasing peptide (BN/GRP) suppress the expression of VEGF, bFGF, and receptors of the EGF/HER family in PC-3 and DU-145 human androgen-independent prostate cancers. AB - BACKGROUND: Antagonists of growth hormone releasing hormone (GHRH) as well as antagonists of bombesin/gastrin releasing peptide (BN/GRP) inhibit the growth of various malignancies (cancers) including prostate cancer. METHODS: We investigated the effects of GHRH antagonists MZ-J-7-118 and RC-J-29-18, BN/GRP antagonists RC-3940-II and RC-3940-Et and the combination of MZ-J-7-118 and RC 3940-II on the growth of PC-3 and DU-145 human androgen independent prostate cancers xenografted s.c. into nude mice. To elucidate the mechanisms of action of these analogs, growth factors like IGF-II (insulin-like growth factor-II), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and epidermal growth factor receptor/human epidermal growth factor receptor (EGF R/HER) family were measured in tumors as well as IGF-I in serum. RESULTS: Antagonists of GHRH and BN/GRP alone or in combination significantly inhibited growth of PC-3 and DU-145 tumors, the greatest inhibition of tumor volume being achieved by combination of MZ-J-7-118 (5 microg/day) and RC-3940-II (10 microg/day). BN/GRP and GHRH antagonists and their combination also decreased the expression of VEGF significantly in PC-3 and non-significantly in DU-145, as measured by radioimmunoassay for VEGF protein and RT-PCR for mRNA levels of VEGF. GHRH and BN/GRP antagonists reduced bFGF concentrations and the maximal binding capacity of EGF receptors, and their mRNA levels in PC-3 and DU-145 tumors. mRNA levels for HER-2 and -3 were also diminished in PC-3 tumors by GHRH and BN/GRP antagonists. No changes in HER-4 were found after treatment. Serum IGF-I and tumoral IGF-II levels were not affected by the analogs. CONCLUSIONS: BN/GRP and GHRH antagonists inhibit growth of PC-3 and DU-145 prostate cancers by suppressing the expression of tumoral growth factors such as VEGF and bFGF as well as the receptors for EGF and related HER-2 and -3. Additive effects on tumor inhibition (TI) in vivo, but not on VEGF, bFGF, or members of the EGF/HER receptor family, can be achieved by the joint administration of both classes of analogs. PMID- 15754344 TI - Anticancer effect of hyperthermia on prostate cancer mediated by magnetite cationic liposomes and immune-response induction in transplanted syngeneic rats. AB - BACKGROUND: The hyperthermic effect of magnetic particles was examined in rat prostate cancer in vivo. Magnetic cationic liposomes (MCLs) have a positive surface charge and generate heat in an alternating magnetic field (AMF) due to hysteresis losses. METHODS: Rat prostate cancer cells (PLS 10; androgen independent) were injected subcutaneously into the flank of F344 rats. MCLs were injected into rat prostate cancer nodules that had grown to 5-6 mm in diameter, and were then exposed to an AMF. Tumor growth rates were measured. To examine whether hyperthermia caused immune induction for PLS 10, cytotoxicity assays and immunohistochemical staining for CD3, CD4, CD8, and Heat Shock Protein (HSP) 70 were performed. RESULT: The tumor temperature increased to 45 degrees C whereas the body temperature remained at around 38 degrees C. Tumor regression was observed in the hyperthermic group. CD3, CD4, and CD8 immunocytes were present in the tumor tissues of the rats exposed to hyperthermia, but they were not detected in any of the tumor tissue of untreated rats. HSP70 also appeared in the viable area at its boundary with the necrotic area. The cytotoxic activity of tumor transplanted rats for PLS 10 cells increased in hyperthermic-treatment rats. CONCLUSION: These results suggest that hyperthermia using MCLs is an effective therapy for prostate cancer, since this treatment appears to kill the prostate cancer cells not only directly by heating but also by inducing an immune response. This therapy may cure not only the primary lesion but also metastatic lesions. PMID- 15754345 TI - Obtaining fresh prostate cancer tissue for research: a novel biopsy needle and sampling technique for radical prostatectomy specimens. AB - BACKGROUND: Fresh or fresh-frozen tissue samples are preferred for molecular profiling as formalin fixation degrades intracellular nucleic acids. Radical prostatectomy (RP) specimens are a valuable source of prostate cancer tissue, but the reliance on whole-organ pathological processing for prognostication limits sampling opportunities. Few studies have addressed specific harvesting techniques using prostatectomy specimens. MATERIALS AND METHODS: Ex vivo biopsies were performed on 23 consecutive fresh RP specimens using a purpose-designed needle. A standard sextant approach was used with an additional lateral biopsy on each side. Cores from each lobe were snap-frozen together and sections assessed by a pathologist blinded to the RP and pre-operative biopsy pathology. Comparison with pre-operative biopsies was performed using the t-test and chi(2) statistical tests. Eleven randomly selected RP specimens were further evaluated for the effects of needle tracks and margin perforation. RESULTS: Cancer was detected in 19 of 23 specimens, giving a sensitivity of 83.6%. The average tumor involvement was 28.3% per section compared with 15.6% for pre-operative biopsies (P < 0.02). There was no statistically significant difference between the groups for either Gleason sum score concordance or tumor location concordance. In 3 of 11 cases, needle margin perforation was identified; in none of the cases did it compromise pathological assessment, although in one case a deeper block resection was required. CONCLUSIONS: Ex vivo biopsy is a useful technique for retrieving fresh tissue whilst preserving organ morphology in RP specimens. The purpose-designed needle and harvesting technique provide good yields of cancer tissue from a high proportion of sampled prostatectomy specimens. PMID- 15754346 TI - Small G-protein RhoE is underexpressed in prostate cancer and induces cell cycle arrest and apoptosis. AB - BACKGROUND: RhoE/Rnd3, a recently described novel member of the Rho GTPases family, was discussed as a possible antagonist of the RhoA protein that stimulates cell cycle progression and is overexpressed and/or overactivated in prostate cancer. We investigated the expression of RhoE and its role in cell cycle regulation and apoptosis in the human prostate. METHODS: RhoE expression in cell lines and tissue specimens was assessed by immunoblot analysis, real-time PCR (RT-PCR), and immunohistochemistry. To elucidate RhoE effects on the prostate, RhoE was cloned and overexpressed in DU-145 prostate cancer. Cell cycle modulation and apoptosis was investigated by immunoblot and FACS analysis. RESULTS: Immunoblot analysis showed a strong RhoE signal in both, benign epithelial and stromal cells. In contrast, almost no protein was detected in various prostate cancer cells. On RT-PCR and microarray analysis, RhoE mRNA expression was significantly reduced in malignant tissue when compared to benign samples. RhoE immunostaining was strong in benign tissue, especially in prostate epithelial cells, whereas it was minimal or absent in malignant tissue. Forced RhoE overexpression in a prostate cancer cell line inhibits the expression of two proteins essential for G2/M transition, namely CDC2 and cyclin B1, and induces G2/M arrest. In addition, apoptotic cell death as measured by a cleavage product of caspase 3 is significantly increased in RhoE-overexpressing cells. CONCLUSION: In conclusion, our findings suggest RhoE as a tumor suppressor gene that is downregulated early in the development of prostate cancer. PMID- 15754347 TI - Dendritic cells generated from patients with androgen-independent prostate cancer are not impaired in migration and T-cell stimulation. AB - BACKGROUND: Dendritic cell (DC)-based vaccination has been investigated as immunotherapy for several types of cancer. A potential drawback to vaccination with autologous monocyte-derived DCs (MoDCs) could be that MoDCs from patients are functionally impaired. In case of androgen-independent prostate cancer (CaP), the cancer itself, diverse prior therapies, and the hormone manipulation may affect the immune system. METHODS: MoDCs from patients suffering from androgen independent CaP were generated according to a clinically applicable protocol to evaluate the phenotype, maturation capacity, migration, and T-cell stimulation of these cells compared with those generated from tumor-free donors. RESULTS: MoDCs generated from CaP patients could be fully matured and efficiently migrated towards the chemokine CCL21. They had a strong potency to activate allogeneic CD4+ and CD8+ T-cells and to present antigens to specific CTL. CONCLUSIONS: Our data suggest that MoDCs from patients with androgen-independent CaP are functionally intact and hence qualify as cellular vaccines for immunotherapy of advanced stage CaP. PMID- 15754348 TI - Population-based risk assessment for other cancers in relatives of hereditary prostate cancer (HPC) cases. AB - BACKGROUND: To identify associations of other cancers with hereditary prostate cancer (HPC) we estimated relative risks (RRs) of 36 different cancers in relatives of prostate cancer cases in the Utah Population Data Base (UPDB), which combines genealogical and cancer data for Utah. METHODS: We utilized known genetic relationships between prostate cancer cases and their relatives with cancer, combined with age- and sex-specific cancer rates calculated internally from the UPDB, to estimate RRs for cancer in relatives of prostate cancer cases. RESULTS: Multiple other cancers were observed in excess in both first- and second degree relatives of HPC cases including colon cancer, non-Hodgkins lymphoma, multiple myeloma, rectal cancer, cancer of the gallbladder, and melanoma (skin). CONCLUSIONS: This analysis supports the existence of heritable prostate cancer syndromes that include other cancers. We hypothesize that the study of homogeneous pedigrees co-segregating prostate cancer and another cancer could allow more straightforward localization and identification of the gene(s) responsible. PMID- 15754349 TI - Aurora-A over-expression in high-grade PIN lesions and prostate cancer. AB - BACKGROUND: Over-expression of Aurora-A (Aurora 2 kinase, STK-15), a protein found in centrosomes thought to be associated with genetic instability, has been previously documented in prostate cancer [Pihan et al.: Cancer Res 61(5):2212 2219, 2001]. It is unknown if this protein is also over-expressed in high-grade prostatic intraepithelial neoplasia (PIN) lesions. METHODS: PIN lesions were examined for increased Aurora-A using immunohistochemical staining on archival paraffin embedded prostatectomy tissue. Aurora-A expression was scored using size, number, and staining intensity. Protein expression was examined and compared between stromal cells, normal glands, high-grade PIN lesions, and invasive cancer. RESULTS: Immunohistochemistry shows an increased expression of Aurora-A in 96% of high-grade PIN cases, and 98% in cancer lesions. Twenty-nine percent of cases of normal glands from cancerous prostates also showed increased Aurora-A expression. CONCLUSIONS: Over-expression of Aurora-A is present in some normal and the majority of high-grade PIN lesions indicating that this may be an early event that leads to the genetic instability seen in prostate carcinogenesis. PMID- 15754350 TI - Androgen receptor signaling and vitamin D receptor action in prostate cancer cells. AB - BACKGROUND: 1,25(OH)2D3 inhibits the growth of prostate cancer cells; previous reports suggest that 1,25(OH)2D3 actions in LNCaP prostate cancer cells are androgen dependent. This is due in part to the observation that the androgen receptor (AR) antagonist, Casodex, modestly inhibits LNCaP cell growth, but reduces the greater growth inhibition induced by 1,25(OH)2D3 to the level of Casodex alone. Because androgen ablation therapy is used for metastatic prostate cancer, we sought to better characterize this androgen dependence. METHODS: We have assessed the requirement for endogenous androgens in 1,25(OH)2D3 mediated growth inhibition of AR+ prostate cancer cell lines. We have also sought the mechanism for anti-androgen mediated reversal of 1,25(OH)2D3 dependent growth inhibition in LNCaP cells. RESULTS: Although 1,25(OH)2D3 does not inhibit the growth of LNCaP cells grown in medium lacking androgens, we find that growth of androgen independent derivatives of LNCaP cells is inhibited by 1,25(OH)2D3. Despite this independence, Casodex treatment reduced the response of these cells to 1,25(OH)2D3 suggesting a unique function for Casodex-bound AR. Because Casodex does not directly inhibit the transcriptional activity of the vitamin D receptor (VDR) we sought a common primary target of VDR and AR action whose VDR dependent transcription could be repressed by Casodex. We report that AS3 (APRIN), a novel gene required for androgen dependent growth arrest, is a primary target for 1,25(OH)2D3 and androgens. Moreover, Casodex reduces induction of AS3 by 1,25(OH)2D3 suggesting that it is a candidate for the Casodex effect. Analysis of functional interactions between AR and VDR in other AR containing prostate cancer cells lines (PC-3 AR, LAPC-4, and 22Rv1) revealed that Casodex reversal was unique to LNCaP derived cells. CONCLUSION: Anti-androgen mediated reversal of 1,25(OH)2D3 dependent growth inhibition is limited to LNCaP derived prostate cancer cell lines. Moreover, the growth of androgen independent derivatives of LNCaP cells in medium depleted of androgens is strongly inhibited by 1,25D. Therefore, most forms of androgen ablation should not eliminate the utility of VDR agonist treatment in most prostate cancers. PMID- 15754351 TI - Genome-wide screen for prostate cancer susceptibility genes in men with clinically significant disease. AB - BACKGROUND: One of the difficulties confronting genetic studies of prostate cancer is the complex and heterogeneous etiology. Given the high population frequency of lesions meeting the histological definition of prostate cancer, a significant portion of men with a positive family history may be diagnosed due to increased surveillance and associated higher likelihood of biopsy. Over diagnosis decreases power to detect genes that increase susceptibility to a clinically significant prostate cancer. METHODS: We re-evaluated all 623 men with prostate cancer in our 188 hereditary prostate cancer families and identified a subset of 244 men with more aggressive disease based upon meeting at least one of the following clinical and/or pathologic criteria: tumor grade Gleason score > or = 7, tumor stage T2c or higher, pretreatment PSA > or = 20 ng/ml, rising PSA after treatment, evidence of metastasis, or death from prostate cancer. RESULTS: Genome wide screens were re-performed by defining men as affected only if they met the criteria for clinically significant disease. The new analyses identified stronger evidence for linkage in Xq27-28 and 22q, as well as several novel loci, including 3p and 9p. CONCLUSIONS: Although, these results need to be confirmed in independent studies, our approach represents an important step to overcome the impact of over diagnosis in genetic studies of prostate cancer. Larger studies that incorporate this approach are needed. PMID- 15754353 TI - Rapid array-based genomic characterization of a subtle structural abnormality: a patient with psychosis and der(18)t(5;18)(p14.1;p11.23). AB - Array-based copy number analysis has recently emerged as a rapid means of mapping complex and/or subtle chromosomal abnormalities. We have compared two such techniques, using bacterial artificial chromosome (BAC) and single nucleotide polymorphism (SNP) arrays in the evaluation of a 45-year-old woman with dysmorphic features, mental retardation, psychosis, and an unbalanced derivative chromosome 18, (46,XX, der(18)t(18;?)(p12;?)). Both array-based methods demonstrated that the additional material on chromosome 18 was of 5p origin. The 5p duplication mapped telomeric to 25.320 Mb (BAC array) and 25.607 Mb (SNP array), corresponding to the band 5p14.1. Both BAC and SNP arrays also showed a deletion involving chromosome 18p extending telomeric from 8.437 Mb (BAC array) and 8.352 Mb (SNP array), corresponding to the band 18p11.23. Molecular cytogenetic mapping using fluorescence in situ hybridization (FISH) supported the array findings and further refined the breakpoint regions, confirming that the BAC and SNP chips were both useful in this regard. Both case reports and linkage analyses have implicated these chromosomal intervals in psychosis. The array based experiments were completed over the course of several days. While these methods do not eliminate the requirement for traditional fine-mapping, they provide an efficient approach to identifying the origin and extent of deleted and duplicated material in chromosomal rearrangements. PMID- 15754354 TI - Differentiating campomelic dysplasia from Cumming syndrome. PMID- 15754355 TI - Fibular aplasia, tibial campomelia, and oligosyndactyly in a male newborn infant: a case report and review of the literature. AB - We report on a male newborn with a rarely described congenital limb deficiency syndrome consisting of shortening and anterior bowing of the right lower limb at the distal third of the tibia with associated overlying soft tissue dimpling, oligodactyly of the right foot, and a left-sided oligosyndactyly of the hand. The right hand and left lower limb were clinically normal. Radiographic examination revealed complete absence of the right fibula, absence of the right-sided Vth ray, and anterior bowing and shortening of the right-sided tibia. Femora, humeri, ulnae, and radii were normal. The infant had neither facial dysmorphia nor other associated anomalies. A limb deficiency syndrome comparable to this case has been reported in a female by Hecht and Scott, the only report classified under OMIM 246570 so far. We found two other reports describing three cases comparable to our case and the female reported by Hecht and Scott, and reviewed these cases. The major common findings in all the five cases consist of fibular aplasia, tibial campomelia, and oligosyndactyly. Therefore, we propose to name it fibular aplasia-tibial campomelia-oligosyndactyly (FATCO) syndrome. Additional case reports are needed for further delineation of this rare limb deficiency syndrome. PMID- 15754356 TI - Gastric juvenile polyposis associated with germline SMAD4 mutation. AB - We treated a 39-year-old woman with hypoproteinemia and anemia who had profuse gastric polyposis. Radiographic and endoscopic examination showed numerous polyps restricted to the stomach. The patient had pulmonary arteriovenous malformations in the left lung. Histological examination of the resected stomach revealed the gastric polyposis to be composed of cystic dilatation of the glands with small areas of adenocarcinoma. These findings were compatible with gastric juvenile polyposis (GJP) accompanied by gastric cancer. Analysis of genomic DNA revealed that the patient had truncating mutation of SMAD4, a responsible gene for juvenile polyposis (JP). Our case suggests that SMAD4 is possibly a responsible gene for GJP. PMID- 15754357 TI - Halteria grandinella: a rapid swimming ciliate with a high frequency of ciliary beating. AB - A ciliated protozoan, Halteria grandinella, swam backward rapidly with a migration distance per second attaining 100 times the cell size. This high swimming velocity was accompanied by a high frequency of ciliary beating. Recordings with a high-speed digital video (10(3) frames/s) revealed that the frequency during forward and backward swimming was, respectively, 105 +/- 10 Hz and 260 +/- 30 Hz. These frequencies are the highest among cilia and flagella reported to date. Electron microscopic observation of the ciliary structure confirmed normal 9 + 2 arrangements of the axoneme except that cilia for migration are bundled into membranelles. Ciliary beating of saponin-treated cells was reactivated by the addition of Mg2+ -ATP, although the beating amplitude was smaller than that of intact cells. Kinetic analysis of the ATP-dependent increase of beating frequency revealed that the maximal frequency in the presence of free Ca2+ and 0.9 microM Ca2+ was approximately 60 and 110 Hz, respectively. A possible mechanism to increase beating frequency with Ca2+ is discussed. PMID- 15754358 TI - Characterization of myosin-II binding to Golgi stacks in vitro. AB - In addition to important roles near the actin-rich cell cortex, ample evidence indicates that multiple myosins are also involved in membrane movements in the endomembrane system. Nonmuscle myosin-II has been shown to have roles in anterograde and retrograde trafficking at the Golgi. Myosin-II is present on Golgi stacks isolated from intestinal epithelial cells and has been localized to the Golgi in several polarized and unpolarized cell lines. An understanding of roles of myosin-II in Golgi physiology will be facilitated by understanding the molecular arrangement of myosin-II at the Golgi. Salt-washing removes endogenous myosin-II from isolated Golgi and purified brush border myosin-II can bind in vitro. Brush border myosin-II binds to a tightly bound Golgi peripheral membrane protein with a K(1/2) of 75 nM and binding is saturated at 0.7 pmol myosin/microg Golgi. Binding studies using papain cleavage fragments of brush border myosin-II show that the 120-kDa rod domain, but not the head domain, of myosin heavy chain can bind directly to Golgi stacks. The 120-kDa domain does not bind to Golgi membranes when phosphorylated in vitro with casein kinase-II. These results suggest that phosphorylation in the rod domain may regulate the binding and/or release of myosin-II from the Golgi. These data support a model in which myosin II is tethered to the Golgi membrane by its tail and actin filaments by its head. Thus, translocation along actin filaments may extend Golgi membrane tubules and/or vesicles away from the Golgi complex. PMID- 15754360 TI - Plasmodium falciparum myosins: transcription and translation during asexual parasite development. AB - Six myosins genes are now annotated in the Plasmodium falciparum Genome Project. Malaria myosins have been named alphabetically; accordingly, we refer to the two latest additions as Pfmyo-E and Pfmyo-F. Both new myosins contain regions characteristic of the functional motor domain of "true" myosins and, unusually for P. falciparum myosins, Pfmyo-F encodes two consensus IQ light chain-binding motifs. Phylogenetic analysis of the 17 currently known apicomplexan myosins together with one representative of each myosin class clusters all but one of the apicomplexan sequences together in Class XIV. This refines the earlier definition of the Class XIV Subclasses XIVa and XIVb. RT-PCR on blood stage parasite mRNA amplifies a specific product for all six myosins and each shows developmentally regulated transcription. Thus: Pfmyo-A and Pfmyo-B genes are transcribed throughout development; Pfmyo-C is predominant in trophozoites; Pfmyo-D occurs in trophozoites and schizonts; Pfmyo-E though barely present in earlier stages is abundant in schizonts; Pfmyo-F increases steadily throughout development and maturation. It is known that Pfmyo-A and Pfmyo-B are synthesised during late schizogony and we now show that Pfmyo-D expression is also temporally regulated to late trophozoites and schizonts where it distributes close to segregating nuclei. Thus, in asexual stages myosin synthesis does not always parallel transcript accumulation, showing that translation is also regulated. The implication is that the mRNAs are either subjected to turnover, synthesised and degraded, or that they are sequestered in an inactivate form until required for protein synthesis. PMID- 15754361 TI - Fine-needle aspiration cytology of pleomorphic hyalinized angiectatic tumor: A case report. AB - Pleomorphic hyalinized angiectatic tumor (PHAT) of soft parts is a neoplasm characterized by spindle and pleomorphic cells associated with an angiectatic vasculature. We describe the cytological findings of a fine-needle aspiration biopsy (FNAB) from the right medial knee of a 45-yr-old woman. The aspirate material was entirely submitted in Cytolit solution. The specimen was moderately cellular and was comprised of spindle cells in a background of fibrinous material. The cells varied from small, bland spindle cells with a fine chromatin pattern and inconspicuous nucleoli to larger pleomorphic cells with coarser chromatin and occasional intranuclear inclusions. Most of the cells were arranged singly with sporadic small cluster formation with indistinct cell borders. Rare mononuclear inflammatory cells morphologically compatible with mast cells were identified. The differential diagnosis include solitary fibrous tumor (SFT) and ancient schwannoma, which also shows fibrous-like material and spindle cells that may have intranuclear inclusions. PMID- 15754359 TI - The 22q11.2 deletion in African-American patients: an underdiagnosed population? AB - Findings associated with the 22q11.2 deletion often include congenital heart malformations, palatal anomalies, immunodeficiency, hypocalcemia, and developmental delay or learning disabilities. Often the clinical suspicion of the diagnosis in a patient with one or more of these findings is heightened based on the presence of a characteristic facial appearance. In our large cohort of 370 patients with the 22q11.2 deletion, we report the under-representation of African Americans in our group, as well as, the paucity of craniofacial dysmorphism in these patients. We note that the absence of the typical facial features may result in decreased ascertainment in this population and, furthermore, may delay the implementation of palliative care, cognitive remediation, and recurrence risk counseling. We, therefore, suggest that the clinician's threshold of suspicion should be lower in African-American patients. PMID- 15754362 TI - Fine-needle aspiration cytology of bilateral mammary fibroadenoma with multinucleated stromal giant cells. PMID- 15754363 TI - Freshwater microorganisms and other arthropods in Papanicolaou smears. AB - Several types of contaminants have been reported in cytopathology. The purpose of this study was to show unusual contaminants in cytological smears, such as freshwater microorganisms and other arthropods. In different routine Papanicolaou smears (sputum, cervicovaginal, and voided urine) we mainly found fragments of mites and insects, caused by an intrinsic contamination of the samples or an extrinsic contamination of the smears. PMID- 15754364 TI - Fine-needle aspiration biopsy of cystic pleomorphic adenoma with adnexa-like differentiation mimicking mucoepidermoid carcinoma: a case report. AB - Although pleomorphic adenomas (PAs) usually can be diagnosed very accurately with fine-needle aspiration biopsy (FNAB), even this common salivary gland neoplasm can be diagnostically challenging and cause pitfalls in cytodiagnosis. In particular, the presence of either cystic changes or squamous, mucinous, or sebaceous metaplasia can lead to a false positive diagnosis of mucoepidermoid carcinoma (MEC). Here, we present a case of a 70-yr-old man with an asymptomatic left deep lobe parotid mass for which CT-guided FNAB was performed. The FNAB cytology revealed cohesive clusters of squamous epithelial cells, sebaceous cells, oncocytes, macrophages, and rare myoepithelial cells. Characteristic metachromatic fibrillar chondromyxoid stroma, which usually is seen in PAs, was not seen in the aspirate. Although cytodiagnosis of PAs was suggested based on the presence of other cellular components, resection was recommended. The subsequent parotidectomy specimen revealed an encapsulated cystic PA with mixed appendageal differentiation including areas of squamous, mucinous, sebaceous, and oncocytic metaplasia. Chondromyxoid stroma was only focally present. Presence of squamous, mucinous, and/or sebaceous metaplasia, especially in the absence of chondromyxoid stroma, presents the potential for misinterpretation of the FNAB as indicative of malignancy in general and MEC in particular. PMID- 15754365 TI - p63 immunostaining in destained bronchoscopic cytological specimens. AB - The p53 homologous squamous stem-cell regulatory protein p63 is expressed in squamous carcinomas but is not characteristically detected in small-cell carcinomas (SCCs). A panel of thyroid transcription factor (TTF) 1 and p63 has been shown to be useful in distinguishing SCCs from poorly differentiated squamous carcinoma of the lung (PDSLC) in small biopsies and cytological cell blocks. Because tumor samples frequently are limited to cytological smears, we attempted to detect p63 in destained slides from a spectrum of pulmonary malignancies. Archival alcohol-fixed smears from 60 cases of cytologically diagnosed malignancies in bronchoscopically (n = 59) or fine-needle aspiration obtained specimens (n = 1) were destained in acid alcohol, postfixed in 10% formalin, subjected to citrate-based antigen retrieval, and immunostained by exposure to anti-p63 monoclonal antibody 4A4, followed by reagents from a streptavidin-biotin immunoperoxidase kit, and diaminobenzidine as the chromogen. Postfixation in 10% formalin was found to be necessary for immunostaining. Normal ciliated and goblet cells were p63 negative, but reserve cells were p63 positive. All cases of squamous-cell carcinoma were positive for p63. Of 10 tumor samples originally diagnosed as SCC, only 6 samples were p63 negative and 4 samples exhibited positive staining. However, proper interpretation of the immunohistochemical (IHC) staining pattern and careful scrutiny of the cytological features and biopsy specimens in three of four cases led us to reclassify three cases into PDSLC. All adenocarcinomas (ACAs; n = 12), large-cell carcinomas (n = 4), and metastatic ACAs (n = 5) were p63 negative. Positive staining was seen in 9/16 tumors designated as non-SCCs; these tumors were not classified further into distinct histological categories.p63 staining in destained slides may be of value in facilitating the differential diagnosis between PDSLC and SCC. Criteria for conservative interpretation of results are discussed and include examination of reserve cells and ciliated cells on the same slide as internal positive and negative controls. PMID- 15754367 TI - Clear-cell carcinoma of the pancreas in fine-needle aspirate. PMID- 15754366 TI - Cytological diagnosis of endocrine tumors of the pancreas by endoscopic ultrasound-guided fine-needle aspiration biopsy. AB - The aim of this study was to evaluate the efficacy and accuracy of endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) in the diagnosis of pancreatic endocrine tumors and to analyze their cytomorphology. Between March 1999 and June 2004, a total of 30 patients with a cytological diagnosis of pancreatic endocrine tumors were found. Their records were retrieved and the cytological materials were analyzed. The mean size of the tumors assessed by EUS was 3.0 cm. Immediate preliminary interpretation was rendered after an average of 1.5 passes. Based on the cellular patterns, cases were divided into three categories: loosely cohesive aggregates, discohesive single cells, and cohesive flat sheets. Most tumor cells had abundant cytoplasm and eccentric nuclei. Chromatin was fine or coarse but was evenly distributed in all cases. Nuclear pleomorphism, multinucleation, intranuclear inclusions, mitotic figures, and necrosis were seen. Immunohistochemical (IHC) studies on cell blocks confirmed the diagnosis in all cases. EUS-guided FNA is efficient and accurate in establishing the diagnosis of pancreatic endocrine tumors. The variety of cellular patterns presents several differential diagnostic issues that should be considered to avoid erroneous interpretation. PMID- 15754368 TI - Fine-needle aspiration cytology of extramammary neoplasms metastatic to the breast. AB - Fine-needle aspiration (FNA) biopsy is the first-line investigation in any breast lump and hence cytomorphological recognition of nonmammary metastatic tumors to the breast and their distinction from primary tumors is important. Metastatic breast neoplasms diagnosed over a 6-yr period from 1997 to 2002 were retrieved from the database of the Department of Cytopathology and the clinical, cytopathological, histochemical, and immunohistochemical findings were correlated with the histopathology of the primary tumor. Fifteen cases of metastatic breast neoplasms were encountered constituting 1.47% of all malignant tumors of the breast diagnosed on FNA. There were 14 female patients and one male patient aged 13-80 yr. The preaspiration clinical diagnosis was either a benign breast lump or a malignancy (primary vs. metastatic). The breast lump was the initial presentation in four cases and the cytodiagnosis of a metastatic malignancy lead to the subsequent detection of the primary malignancy. These included one case each of melanoma, myeloma, rhabdomyosarcoma, and small-cell carcinoma of the lung. There were five pediatric cases that included four cases of rhabdomyosarcoma and one case of leukemic deposit. The adult cases included two cases each of melanoma, small-cell carcinoma, and myeloma; one case of choriocarcinoma; and three cases of soft-tissue sarcomas. These included two cases of malignant fibrous histiocytoma (MFH) and one case of leiomyosarcoma. The presence of unusual cytomorphological patterns on breast FNA should alert the cytopathologist to the possibility of a metastatic breast neoplasm, even if not suspected clinically. A detailed history of the patient, clinical correlation, and immunocytochemistry helps in establishing an accurate diagnosis, which avoids unnecessary surgery and ensures appropriate treatment. PMID- 15754369 TI - Fine-needle aspiration cytology of metastatic nasopharyngeal carcinoma. AB - Cytological features of nasopharyngeal carcinoma (NPC) were reviewed in an attempt to select cytological criteria that permit a specific recognition of metastases. For this purpose, 54 fine-needle aspiration (FNA) procedures from 43 patients with NPC were analyzed. Thirty-two (59.3%) procedures were performed before the histological diagnosis. In 25 (46.3%) procedures, smears showed many neoplastic single cells, clusters, and abundant lymphoid cells (mixed pattern). A dissociated (single cell) pattern consisting of individual neoplastic and lymphoid cells was seen in 18 (33.3%) cases. Finally, 11 (20.4%) cases showed cohesive epithelial clusters (cohesive pattern) without relevant cellular dissociation or lymphoid cells. Squamous-cell differentiation was seen in three of these cases. Most single neoplastic cells presented as large, pleomorphic naked nuclei. Other interesting findings were granulomas (n = 3), prominent eosinophilic infiltrates (n = 4), and suppurative changes (n = 5). In most smears with mixed and dissociated patterns, a nasopharyngeal origin could be suggested. On the contrary, those smears with a cohesive pattern were indistinguishable from other head and neck carcinomas. The presence (on cervical lymph nodes) of a dissociated or mixed (single cells and groups) architectural pattern of large, anaplastic cells and naked nuclei accompanied by an abundant lymphoid component is highly suggestive of undifferentiated NPC. Cytology offers a rapid diagnosis, establishes the necessity of a complete cavum examination, and helps in avoiding unnecessary and harmful biopsies. PMID- 15754370 TI - Atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion: the practice experience of a hospital-based reference laboratory with this new Bethesda system diagnostic category. AB - The 2001 Bethesda System diagnostic category of atypical squamous cells cannot exclude a high grade squamous intraepithelial lesion (ASC-H) was developed and implemented after many studies that proved its clinical relevance. In this study, we describe the practice experience of a hospital-based reference laboratory with this new diagnostic category. The anatomic pathology computer database was searched, and 414 Papanicolaou (Pap) smears signed out as ASC-H were discovered among 60,390 Pap smear accessions over a 17-month period of time. One hundred four cases had corresponding tissue specimens. Slides from all Pap smears and tissue specimens were reviewed, and five Pap smears were reclassified, leaving 99 study cases. In our laboratory, 88.9% of the study cases had either low or high grade dysplasia diagnosed on subsequent tissue specimens. The positive predictive value of ASC-H for histologically proven high-grade squamous intraepithelial lesions (HSIL's) was 40.4%. Of those having human papillomavirus (HPV) hybrid capture II testing, high-risk HPV types were detected in 73.9% of cases. The majority of study cases had less than 25 atypical cells. In two hysterectomy cases and three loop electrosurgical excession procedure (LEEP) conization cases, high-grade dysplasia was present as a single microscopic focus, suggesting that the paucity of atypical cells in ASC-H Pap smears may be secondary to small lesion sampling. Thirteen study patients were postmenopausal and 30.8% had low grade dysplasia, and of these, 46.2% had high-grade dysplasia on subsequent tissue specimens. In conclusion, our practice experience with ASC-H is similar to that reported in the literature before the 2001 Bethesda System. PMID- 15754371 TI - Fine-needle aspiration diagnosis of primary cutaneous cryptococcosis in an immunocompetent patient: a case report. AB - We report a case of a primary cutaneous cryptococcosis (PCC) in immunocompetent man diagnosed by fine-needle aspiration (FNA) and confirmed by special stains and culture study. The patient presented with a 2-wk history of two separate skin nodules over the forehead and the left lumbar region. FNA smears revealed a necrotizing suppurative granulomatous inflammation with numerous yeasts of Cryptococcus surrounded by clear halos, which were present within multinucleated giant cells and in the intercellular spaces. This was confirmed subsequently by special stains and a culture of the aspirated material. This case report reemphasizes the role of FNA as a useful modality in the early diagnosis of cutaneous cryptococcosis, especially in clinically unsuspected cases. PMID- 15754372 TI - Fine-needle aspiration cytology of metastatic transitional cell carcinoma. AB - In this article we described the fine-needle aspiration cytology (FNAC) of five cases of metastatic transitional cell carcinoma (TCC). There were four cases of metastatic lymph nodes and one case of metastatic skin lesion. All of the TCC cases were primarily in the urinary bladder and were high grade on histopathology (grade 3). Three cases showed bladder muscle involvement and two cases showed superficial TCC at the time of primary diagnosis. FNAC smears showed abundant cellularity. The cells were present in discrete and small syncytial clusters. Nuclear position of the cell was central to eccentric. Many cells showed prominent nucleoli. Cercariform cells (CCs) were noted in four cases. These cells are malignant cells with a nucleated globular body and a unipolar nontapering cytoplasmic process. Two cases showed intranuclear inclusions. Prominent cytoplasmic vacuoles were noted in three cases. In addition, cell cannibalism and attempted pearl formations were noted in two cases.In conclusion, clinical history along with the certain cytological features such as the presence of CCs, cells with eccentric nuclei, and intranuclear inclusions are helpful to diagnose metastatic TCC on FNAC material. PMID- 15754373 TI - Cytomorphology and morphometry of small round-cell tumors in the region of the kidney. AB - Small round-cell tumors (SRCTs), with malignant cell components measuring 10 m or less in diameter with scanty cytoplasm in alcohol-fixed smears, pose a diagnostic challenge at fine-needle aspiration cytology (FNAC), especially when they are situated in and around the kidney and need facilities such as electron microscopy, immunohistochemistry, tissue culture, and cytogenetics for their subtyping. A precise cytodiagnosis of SRCTs is important because a definite diagnosis is mandatory in preoperative diagnostic workup for presurgical chemotherapy in these cases. With this view in mind, an attempt has been made to diagnose SRCTs in the region of the kidney based on cytomorphology and morphometry alone so as to facilitate its diagnosis in a simple cytology laboratory of a developing country where facilities for auxiliary techniques are not easily available. Of 2,028 abdominal aspirates in a 12-yr period, 36 SRCTs were diagnosed in the region of the kidney by correlating with histology, radiology, and clinical features. The smears were studied for cellularity, morphology, pattern of cell arrangement, and smear background and morphometrically analyzed using an ocular micrometer. An aspirate with preponderant malignant round cells that were larger or double the size of red blood cells in air-dried smears or measured less than 10 micro in diameter in alcohol-fixed smears was considered as a small blue-cell tumor. Twenty-one were diagnosed as Wilms' tumor (WT), 10 were diagnosed as neuroblastoma (NB), 3 were ganglioneuroblastoma (GNB), 1 was a cellular congenital mesoblastic nephroma (CMN), and 1 was an adrenocortical carcinoma (ACC). Cell clusters with neuropil and cytoplasmic processes were diagnostic of NB, ganglion cells of GNB, and blastema with tubular differentiation in WT. Aspirates from CMN and ACC were considered as simulators/mimickers of SRCT because they had superficial resemblance to SRCT and their differentiating cytomorphological features observed at histology were too subtle to be noted at cytology. The latter were appreciated only on retrospective analysis after histological confirmation.Thus, morphometry in correlation with cytology, clinical history, physical findings, and radiological data is helpful in guided FNA for a definite diagnosis of SRCT in the region of the kidney. One needs to keep in mind the mimickers of small round cell lesions at this anatomic site. PMID- 15754374 TI - Fusarium macroconidia in Pap smears. PMID- 15754375 TI - Fine-needle aspiration of the testis in azoospermic men: the value of measuring serum follicle stimulating hormone and testicular size. AB - Testicular fine-needle aspiration biopsy (FNAB) is used as a primary tool in assessing azoospermic infertile men in our institution. If the FNA is negative for sperm, a subsequent testicular biopsy specimen and wet preparation are obtained with possible immediate fertilization. To our knowledge, the value of these techniques in the context of testicular size and serum levels of follicle stimulating hormone (FSH) has not been explored. We reviewed 453 FNA biopsies of the testis performed for the identification of sperm in infertile azoospermic males between 1999 and 2000. We identified cases that had all three procedures (FNA, wet preparation and biopsy) performed. These were the patients that were compared for testicular size using Seager orchiometry and serum levels of FSH. Of the 453 FNAs reviewed, 128 (21%) had all three procedures performed. Seventy-two cases (56%) were negative in all three tests, 26 (20%) cases were positive in all tests, and 30 cases (23%) had different results. These 30 cases were excluded. The testicular size in the negative group ranged from 4 to 16 ml (median, 10 ml) and the positive group had testicular sizes ranging from 10 to 25 ml (median, 15 ml; P = 0.0001). The negative group had serum FSH levels ranging from 3 to 52 IU/l (median, 19 IU/l) whereas the positive group had serum levels ranging from 3 to 26 IU/l (median, 10 IU/l; P = 0.0001). Our findings suggest that in infertile azoospermic men, a testicular size of <10 ml combined with a serum FSH level of >19 IU/l, the chances of retrieving sperm are minimal using all three diagnostic modalities. The use of this cutoff point would lead to a 30-50% reduction in the number of procedures performed. This reduction would have a significant impact on the management of these patients with a significant decrease in cost, logistics, and patient anxiety. PMID- 15754376 TI - Thyroid fine-needle aspiration biopsy in children and adolescents: experience with 218 aspirates. AB - To evaluate the role of fine-needle aspiration (FNA) biopsy of thyroid nodules in pediatric and adolescent patients, the cytology reports of 218 thyroid FNA biopsies performed on children and adolescents ranging from 10 to 21 yr of age were reviewed. The cytology diagnoses were categorized into four groups: unsatisfactory, benign, suspicious, and malignant. One hundred nineteen (54%) of the aspirates were diagnosed as "benign," 20 (9%) were diagnosed as suspicious for malignancy; and 17 (8%) were diagnosed as malignant. Sixty-two (28%) of the aspirates were read as unsatisfactory for interpretation. Sensitivity of thyroid FNA in diagnosing thyroid malignancy relative to final histological diagnoses was 100%, and specificity was 65%. FNA of thyroid nodules in the pediatric and adolescent population is comparably as sensitive and specific as in the adult population. The acceptance of this procedure in the routine evaluation of young patients' thyroid nodules should reduce the number of unnecessary surgeries for benign thyroid disease. PMID- 15754377 TI - Budesonide combined with UDCA to improve liver histology in primary biliary cirrhosis: a three-year randomized trial. AB - Ursodeoxycholic acid (UDCA) is a safe medical therapy for primary biliary cirrhosis (PBC), but its effect on liver histology remains uncertain. Budesonide is a glucocorticoid with high receptor activity and high first-pass metabolism in liver. We evaluated the combination of budesonide and UDCA on liver histology and compared this with UDCA alone in a 3-year prospective, randomized, open multicenter study. Patients with PBC (n = 77), at stages I to III, were randomized into 2 treatment arms, A (n = 41): budesonide 6 mg/d and UDCA 15 mg/kg/d and B (n = 36): UDCA 15 mg/kg/d. Liver histology was assessed at the beginning and at the end of the study. Liver function tests and glucose and cortisol values were determined every 4 months. Paired liver biopsy specimens were available from 69 patients (A = 37 and B = 32). Stage improved 22% in group A but deteriorated 20% in group B (P = .009). Fibrosis decreased 25% in group A but increased 70% in group B (P = .0009). S-PIIINP decreased significantly in group A. Inflammation decreased in both groups, 34% in group A (P = .02), but only 10% in group B (P = NS). Serum liver enzymes decreased significantly in both treatment arms. Bilirubin values rose in group B but stayed stable in group A (A/B P = .002). A mild systemic glucocorticoid effect from budesonide was evident after 2 years. In conclusion, budesonide combined with UDCA improved liver histology, whereas the effect of UDCA alone was mainly on laboratory values. Studies with longer follow-up using a combination of budesonide and UDCA are warranted to confirm safety and effects. PMID- 15754378 TI - There's madness in the methods. PMID- 15754380 TI - Targeted expression of Cre recombinase in macrophages and osteoclasts in transgenic mice. AB - To develop specific conditional gene ablation in the hematopoietic myeloid osteoclast lineage, transgenic mice expressing Cre recombinase under the control of the CD11b promotor were generated on the C57BL/6 background. The cellular specificity of Cre activity following recombination was quantified in the Z/EG reporter transgenic mice by FACS analysis with lineage-specific markers and EGFP coexpression. A high degree of recombination, as evidenced by EGFP-positive cells, was demonstrated in macrophages and granulocytes of bone marrow and spleen by the presence of double-positive cells CD11b/EGFP and Gr1/EGFP, respectively. Interestingly, the peritoneal macrophage population showed almost complete DNA recombination at large. Most important, mature osteoclast cells derived from the double transgenic bone marrow and spleen progenitors were EGFP-positive. Hence, these CD11b-Cre mice will provide a unique tool to unravel novel gene function and activities involved during osteoclast and macrophage differentiation and maturation processes. PMID- 15754382 TI - Two-dimensional electrophoresis (2-DE). PMID- 15754381 TI - Cross-species sperm-FISH assays for chemical testing and assessing paternal risk for chromosomally abnormal pregnancies. AB - The father, like the mother, can transmit genetic defects to his offspring that are detrimental for normal development and a healthy life. Epidemiological studies have identified associations between several paternal exposures and abnormal reproductive outcomes, but these types of studies are inherently complex and expensive, and the risk factors for the paternal contribution to abnormal reproductive outcomes remain poorly understood. Several sensitive methods have been developed for detecting mutations and chromosomal damage directly in sperm. These assays are potential bioindicators for paternal risk factors for infertility, spontaneous abortions, aneuploidy syndromes, and genetic diseases in children. Among these methods, fluorescence in situ hybridization (FISH) has been adapted for the detection of numerical and structural chromosomal abnormalities in the sperm of an expanding number of species, including humans and rodents. Sperm FISH has identified several potential paternal risk factors such as age, drugs, lifestyles, and various environmental/occupational exposures. Here, we summarize the status of the development and usage of these sperm-FISH assays and suggest strategies for prioritizing chemical agents for epidemiological investigations to assess paternal risk for abnormal reproductive outcome. PMID- 15754383 TI - A novel microfluidic mixer utilizing electrokinetic driving forces under low switching frequency. AB - This paper presents a novel technique in which low-frequency periodic electrokinetic driving forces are utilized to mix electrolytic fluid samples rapidly and efficiently in a double-T-form microfluidic mixer. Without using any additional equipment to induce flow perturbations, only a single high-voltage power source is required for simultaneously driving and mixing the sample fluids which results in a simple and low-cost system for the mixing purpose. The effectiveness of the mixer as a function of the applied electric field and the periodic switching frequency is characterized by the intensity distribution calculated downstream from the mixing zone. The present numerical and experimental results confirm that the proposed double-T-form micromixer has excellent mixing capabilities. The mixing efficiency can be as high as 95% within a mixing length of 1000 microm downstream from the secondary T-junction when a 100 V/cm driving electric field strength and a 2 Hz periodic switching frequency are applied. The results reveal that the optimal switching frequency depends upon the magnitude of the main applied electrical field. The rapid double-T-form microfluidic mixer using the periodic driving voltage switching model proposed in this study has considerable potential for use in lab-on-a-chip systems. PMID- 15754384 TI - Multicomponent domino processes based on the organocatalytic generation of conjugated acetylides: efficient synthetic manifolds for diversity-oriented molecular construction. AB - The organocatalytic generation of a strong base by the action of a good nucleophile is the base for the in situ catalytic generation of conjugated acetylides in the presence of aldehydes or activated ketones. The method is affordable in a multicomponent, domino format able to generate a chemically diverse set of multifunctionalized adducts that are very well suited for diversity-oriented molecular construction. The domino process involves a nucleophile as catalyst and a terminal conjugated alkyne (H-C[triple chemical bond]C-Z) and an aldehyde or activated ketone as building blocks. The chemical outcome of this process changes dramatically as a function of the nucleophile (tertiary amine or phosphine), temperature, stoichiometry, and solvent. These multicomponent domino processes achieve molecular construction with good atom economy and, very importantly, with an exquisite chemo-differentiating incorporation of identical starting units into the products (nondegenerated chemical output). These properties convert the H-C[triple chemical bond]C-Z unit into a specific building block for diversity-oriented molecular construction. Applications to the modular and diversity-oriented synthesis of relevant heterocycles are discussed. A protocol involving two coupled domino processes linked in a one-pot manner will be discussed as an efficient synthetic manifold for the modular and diversity-oriented construction of multisubstituted nitrogen containing heterocycles. PMID- 15754385 TI - The structure of hormaomycin and one of its all-peptide aza-analogues in solution: syntheses and biological activities of new hormaomycin analogues. AB - Four new aza-analogues of hormaomycin 1, a secondary metabolite with interesting biological activities produced by Streptomyces griseoflavus, were synthesized and subjected to preliminary tests of their antibiotic activity to provide new insights into the structure-activity relationship studies of this class of compounds. The solution structures of hormaomycin 1 and its aza-analogue 2 a were determined by NMR spectroscopy. The data exhibited a reasonably rigid conformation for both molecules, stabilized by stacking interactions between the aromatic moieties attached to the ring and the side chain. According to NMR spectral data the aza-analogue epi-2 a has a rather different conformation and indeed shows no antibacterial activity whatsoever. PMID- 15754386 TI - A modular approach toward block copolymers. AB - A novel methodology for the formation of block copolymers has been developed that combines ring-opening metathesis polymerization (ROMP) with functional chain transfer agents (CTAs) and self-assembly. Telechelic homopolymers of cyclooctene derivatives end-functionalized with hydrogen-bonding or metal-coordination sites are formed through the combination of ROMP with a corresponding functional CTA. These telechelic homopolymers are fashioned with a high control over molecular weight and without the need for post-polymerization procedures. The homopolymers undergo fast and efficient self-assembly with their complement homopolymer or small molecule analogue to form block-copolymer architectures. The block copolymers show equivalent association constants as their small molecule analogues described in the literature, regardless of size or nature of the complementary unit or the polymer side chain. PMID- 15754387 TI - Initial experience from a combination of systemic and regional chemotherapy in the treatment of patients with nonresectable cholangiocellular carcinoma in the liver. AB - AIM: In nonresectable cholangiocellular carcinoma (CCC) therapeutic options are limited. Recently, systemic chemotherapy has shown response rates of up to 30%. Additional regional therapy of the arterially hyper vascularized hepatic tumors might represent a rational approach in an attempt to further improve response and palliation. Hence, a protocol combining transarterial chemoembolization and systemic chemotherapy was applied in patients with CCC limited to the liver. METHODS: Eight patients (6 women, 2 men, mean age 62 years) with nonresectable CCC received systemic chemotherapy (gemcitabine 1 000 mg/m(2)) and additional transarterial chemoembolization procedures (50 mg/m(2) cisplatin, 50 mg/m(2) doxorubicin, up to 600 mg degradable starch microspheres). Clinical follow-up of patients, tumor markers, CT and ultrasound were performed to evaluate maximum response and toxicity. RESULTS: Both systemic and regional therapies were tolerated well; no severe toxicity (WHO III/IV) was encountered. Nausea and fever were the most commonly observed side effects. A progressive rarefication of the intrahepatic arteries limited the maximum number of chemoembolization procedures in 4 patients. A median of 2 chemoembolization cycles (range, 1-3) and a median of 6.5 gemcitabine cycles (range, 4-11) were administered. Complete responses were not achieved. As maximum response, partial responses were achieved in 3 cases, stable diseases in 5 cases. Two patients died from progressive disease after 9 and 10 mo. Six patients are still alive. The current median survival is 12 mo (range, 9-18); the median time to tumor progression is 7 mo (range, 3-18). Seven patients suffered from tumor-related symptoms prior to therapy, 3 of these experienced a treatment-related clinical relief. In one patient the tumor became resectable under therapy and was successfully removed after 10 mo. CONCLUSION: The present results indicate that a combination of systemic gemcitabine therapy and repeated regional chemoembolizations is well tolerated and may enhance the effect of palliation in a selected group of patients with intrahepatic nonresectable CCC. PMID- 15754388 TI - Enhanced migration of tissue inhibitor of metalloproteinase overexpressing hepatoma cells is attributed to gelatinases: relevance to intracellular signaling pathways. AB - AIM: To study the effect of gelatinases (especially MMP-9) on migration of tissue inhibitor of metalloproteinase (TIMP-1) overexpressing hepatoma cells. METHODS: Wild type HepG2 cells, cells stably transfected with TIMP-1 and TIMP-1 antagonist (MMP-9-H401A, a catalytically inactive matrix metalloproteinase (MMP) which still binds and neutralizes TIMP-1) were incubated in Boyden chambers either with or without Galardin (a synthetic inhibitor of MMP-1, -2, -3, -8, -9) or a specific inhibitor of gelatinases. RESULTS: Compared to wild type HepG2 cells, the cells overexpressing TIMP-1 showed 115% migration (P<0.05) and the cells overexpressing MMP-9-H401A showed 62% migration (P<0.01). Galardin reduced cell migration dose dependently in all cases. The gelatinase inhibitor reduced migration in TIMP-1 overexpressing cells predominantly. Furthermore, we examined intracellular signal transduction pathways of TIMP-1-dependent HepG2 cells. TIMP-1 deactivates cell signaling pathways of MMP-2 and MMP-9 involving p38 mitogen-activated protein kinase. Specific blockade of the ERK pathway suppresses gelatinase expression either in the presence or absence of TIMP-1. CONCLUSION: Overexpressing functional TIMP-1- enhanced migration of HepG2-TIMP-1 cells depends on enhanced MMP-activity, especially MMP-9. PMID- 15754389 TI - Expression of cyclooxygenase-2 in colorectal cancer and its clinical significance. AB - AIM: To clarify the clinicopathologic significance of COX-2 expression in human colorectal cancer. METHODS: A total of 128 surgically resected colorectal cancer specimens were immunohistochemically analyzed with the use of anti-COX-2, anti VEGF and anti-MMP-2 antibodies. The relationship between the cyclooxygenase-2 expression in primary lesions of colorectal cancer and clinicopathologic parameters was evaluated by chi-square test. RESULTS: Among 128 cases of colorectal cancer, 87 (67.9%) were positive for cyclooxygenase-2. The expression of cyclooxygenase-2 was significantly correlated with the depth of invasion, stage of disease, and metastasis (lymph node and liver). Patients in T3-T4, stages III-IV and with metastasis had much higher expression of cyclooxygenase-2 than ones in T1-T2, stages I-II and without metastasis (P<0.05). Among 45 cases of colorectal cancer with lymph node metastasis, the COX-2- positive rate was 86.7% (39/45) for primary lesions and diffuse cytoplasmic staining for COX-2 protein was detected in cancer cells in 100% of metastatic lesions of the lymph nodes. VEGF expression was detected in 49 tumors (38.3%), and VEGF expression was closely correlated with COX-2 expression. The positive expression rate of VEGF (81.6%) in the cyclooxygenase-2-positive group was higher than that in the cyclooxygenase-2- negative group (18.4%, P<0.05). MMP-2 expression was detected in 88 tumors (68.8%), and MMP-2 expression was closely correlated with COX-2 expression. The positive expression rate of MMP-2 (79.6%) in the positive COX-2 group was higher than that in the negative COX-2 group (20.4%, P<0.05). CONCLUSION: Cyclooxygenase-2 may be associated with tumor progression by modulating the angiogenesis and cancer cell motility and invasive potential in colorectal cancer and it can be used as a possible biomarker. PMID- 15754390 TI - Importance of adequate immunosuppressive therapy for the recovery of patients with "life-threatening" severe exacerbation of chronic hepatitis B. AB - AIM: Hepatitis B virus (HBV) re-activation often occurs spontaneously or after withdrawal of immunosuppressive therapy in patients with chronic hepatitis B. Severe exacerbation, sometimes developing into fulminant hepatic failure, is at high risk of mortality. The efficacy of corticosteroid therapy in "clinically severe" exacerbation of chronic hepatitis B has not been well demonstrated. In this study we evaluated the efficacy of early introduction of high-dose corticosteroid therapy in patients with life-threatening severe exacerbation of chronic hepatitis B. METHODS: Twenty-two patients, 14 men and 8 women, were defined as "severe" exacerbation of chronic hepatitis B using uniform criteria and enrolled in this study. Eleven patients were treated with corticosteroids at 60 mg or more daily with or without anti-viral drugs within 10 d after the diagnosis of severe disease ("early high-dose" group) and 11 patients were either treated more than 10 d or untreated with corticosteroids ("non-early high-dose" group). RESULTS: Mean age, male-to-female ratio, mean prothrombin time (PT) activity, alanine transaminase (ALT) level, total bilirubin level, positivity of HBeAg, mean IgM-HBc titer, and mean HBV DNA polymerase activity did not differ between the two groups. Ten of 11 patients of the "early high-dose" group survived, while only 2 of 11 patients of the "non-early high-dose" group survived (P<0.001). During the first 2 wk after the introduction of corticosteroids, improvements in PT activities and total bilirubin levels were observed in the "early high-dose" group. Both ALT levels and HBV DNA polymerase levels fell in both groups. CONCLUSION: The introduction of high-dose corticosteroid can reverse deterioration in patients with "clinically life-threatening" severe exacerbation of chronic hepatitis B, when used in the early stage of illness. PMID- 15754391 TI - Effect of endothelin-1 receptor antagonists on histological and ultrastructural changes in the pancreas and trypsinogen activation in the early course of caerulein-induced acute pancreatitis in rats. AB - AIM: To assess the effect of non-selective ET(A/B) (LU 302872) and selective ET(A) (LU 302146) antagonist on pancreatic histology and ultrastructure of acinar cells in connection with trypsinogen activation in early caerulein-induced AP. METHODS: Male Wistar rats with caerulein-induced AP, lasting 4 h, were treated i.p. with 10 and 20 mg/kg b.w. of each antagonist. Edema, inflammatory infiltration, necrosis and vacuolization of acinar cells in the pancreas were scored at 0-3 scale. Free active trypsin (FAT), total potential trypsin (TPT) after activation with enterokinase, and index of trypsinogen activation (%FAT/TPT) were assayed in pancreatic homogenates. RESULTS: In untreated AP, the edema, inflammatory infiltration, necrosis and vacuolization increased as compared to control healthy rats (P<0.01). None of the treatment exerted any meaningful effect on the edema and inflammatory infiltration. The selective antagonist increased slightly the necrosis score to 0.82+/-0.06 at higher dose (P<0.05) vs 0.58+/-0.06 in untreated AP. The non-selective antagonist increased slightly the vacuolization score to 2.41+/-0.07 at higher dose (P<0.01) vs 1.88+/ 0.08 in untreated AP. The decrease in the number of zymogen granules, disorganization of endoplasmic reticulum, autophagosomes and cytoplasmic vacuoles were more prominent in treated AP than in untreated AP groups. %FAT/TPT in untreated AP increased about four times (18.4+/-3.8 vs 4.8+/-1.3 in control group without AP, P<0.001). Treatment of AP with both antagonists did not affect significantly augmented trypsinogen activation. CONCLUSION: The treatment with endothelin-1 receptors (non-selective ET(A/B) and selective ET(A)) antagonists has essential effect neither on the edema and inflammatory infiltration nor on trypsinogen activation observed in the early course of caerulein-induced AP. Nevertheless a slight increase of the necrosis and vacuolization score and some of the ultrastructural data could suggest the possibility of their undesired effects in caerulein-induced AP at investigated doses. PMID- 15754392 TI - Antidiabetic thiazolidinediones induce ductal differentiation but not apoptosis in pancreatic cancer cells. AB - AIM: Thiazolidinediones (TZD) are a new class of oral antidiabetic drugs that have been shown to inhibit growth of same epithelial cancer cells. Although TZD were found to be ligands for peroxisome proliferator-activated receptor gamma (PPARgamma), the mechanism by which TZD exert their anticancer effect is presently unclear. In this study, we analyzed the mechanism by which TZD inhibit growth of human pancreatic carcinoma cell lines in order to evaluate the potential therapeutic use of these drugs in pancreatic adenocarcinoma. METHODS: The effects of TZD in pancreatic cancer cells were assessed in anchorage independent growth assay. Expression of PPARgamma was measured by reverse transcription polymerase chain reaction and confirmed by Western blot analysis. PPARgamma activity was evaluated by transient reporter gene assay. Flow cytometry and DNA fragmentation assay were used to determine the effect of TZD on cell cycle progression and apoptosis respectively. The effect of TZD on ductal differentiation markers was performed by Western blot. RESULTS: Exposure to TZD inhibited colony formation in a PPARgamma-dependent manner. Growth inhibition was linked to G1 phase cell cycle arrest through induction of the ductal differentiation program without any increase of the apoptotic rate. CONCLUSION: TZD treatment in pancreatic cancer cells has potent inhibitory effects on growth by a PPAR-dependent induction of pancreatic ductal differentiation. PMID- 15754393 TI - Spatial organization of bacterial flora in normal and inflamed intestine: a fluorescence in situ hybridization study in mice. AB - AIM: To study the role of intestinal flora in inflammatory bowel disease (IBD). METHODS: The spatial organization of intestinal flora was investigated in normal mice and in two models of murine colitis using fluorescence in situ hybridization. RESULTS: The murine small intestine was nearly bacteria-free. The normal colonic flora was organized in three distinct compartments (crypt, interlaced, and fecal), each with different bacterial compositions. Crypt bacteria were present in the cecum and proximal colon. The fecal compartment was composed of homogeneously mixed bacterial groups that directly contacted the colonic wall in the cecum but were separated from the proximal colonic wall by a dense interlaced layer. Beginning in the middle colon, a mucus gap of growing thickness physically separated all intestinal bacteria from contact with the epithelium. Colonic inflammation was accompanied with a depletion of bacteria within the fecal compartment, a reduced surface area in which feces had direct contact with the colonic wall, increased thickness and spread of the mucus gap, and massive increases of bacterial concentrations in the crypt and interlaced compartments. Adhesive and infiltrative bacteria were observed in inflamed colon only, with dominant Bacteroides species. CONCLUSION: The proximal and distal colons are functionally different organs with respect to the intestinal flora, representing a bioreactor and a segregation device. The highly organized structure of the colonic flora, its specific arrangement in different colonic segments, and its specialized response to inflammatory stimuli indicate that the intestinal flora is an innate part of host immunity that is under complex control. PMID- 15754394 TI - Alterations of mast cells and TGF-beta1 on the silymarin treatment for CCl(4) induced hepatic fibrosis. AB - AIM: Silymarin is a potent antioxidant, antiinflammatory and anti-fibrogenic agent in the liver, which is mediated by alteration of hepatic Kupffer cell function, lipid peroxidation, and collagen production. Especially, in hepatic fibrogenesis, mast cells are expressed in chronic inflammatory conditions, and promote fibroblast growth and stimulate production of the extracellular matrix by hepatic stellate cells. METHODS: We examined the inhibitory mechanism of silymarin on CCl(4)-induced hepatic cirrhosis in rats. At 4, 8, and 12 wk, liver tissues were examined histopathologically for fibrotic changes produced by silymarin treatment. RESULTS: In the silymarin with CCl(4)-treated group, increase of hepatic stellate cells and TGF-beta1 production were lower than in the CCl(4)-treated group at early stages. Additionally, at the late fibrogenic stage, expressions of TGF-beta1 were weaker and especially not expressed in hepatocytes located in peripheral areas. Moreover, the number of mast cell in portal areas gradually increased and was dependent on the fibrogenic stage, but those of CCl(1)+silymarin-treated group decreased significantly. CONCLUSION: Anti fibrotic and antiinflammatory effects of silymarin were associated with activation of hepatic stellate cells through the expression of TGF-beta1 and stabilization of mast cells. These results suggest that silymarin prevent hepatic fibrosis through suppression of inflammation and hypoxia in the hepatic fibrogenesis. PMID- 15754395 TI - Up-regulation of intestinal nuclear factor kappa B and intercellular adhesion molecule-1 following traumatic brain injury in rats. AB - AIM: Nuclear factor kappa B (NF-kappaB) regulates a large number of genes involved in the inflammatory response to critical illnesses, but it is not known if and how NF-kappaB is activated and intercellular adhesion molecule-1 (ICAM-1) expressed in the gut following traumatic brain injury (TBI). The aim of current study was to investigate the temporal pattern of intestinal NF-kappaB activation and ICAM-1 expression following TBI. METHODS: Male Wistar rats were randomly divided into six groups (6 rats in each group) including controls with sham operation and TBI groups at hours 3, 12, 24, and 72, and on d 7. Parietal brain contusion was adopted using weight-dropping method. All rats were decapitated at corresponding time point and mid-jejunum samples were taken. NF-kappaB binding activity in jejunal tissue was measured using EMSA. Immunohistochemistry was used for detection of ICAM-1 expression in jejunal samples. RESULTS: There was a very low NF-kappaB binding activity and little ICAM-1 expression in the gut of control rats after sham surgery. NF-kappaB binding activity in jejunum significantly increased by 160% at 3 h following TBI (P<0.05 vs control), peaked at 72 h (500% increase) and remained elevated on d 7 post-injury by 390% increase. Compared to controls, ICAM-1 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 24 h following TBI and maximally expressed at 72 h post-injury (P<0.001). The endothelial ICAM-1 immunoreactivity in jejunal mucosa still remained strong on d 7 post-injury. The peak of NF-kappaB activation and endothelial ICAM-1 expression coincided in time with the period during which secondary mucosal injury of the gut was also at their culmination following TBI. CONCLUSION: TBI could induce an immediate and persistent up regulation of NF-kappaB activity and subsequent up-regulation of ICAM-1 expression in the intestine. Inflammatory response mediated by increased NF kappaB activation and ICAM-1 expression may play an important role in the pathogenesis of acute gut mucosal injury following TBI. PMID- 15754396 TI - Effects of Guiyuanfang and autologous transplantation of bone marrow stem cells on rats with liver fibrosis. AB - AIM: To investigate the therapeutic effects of Guiyuanfang and bone marrow stem cells (BMSCs) on rats with liver fibrosis. METHODS: Liver fibrosis model was induced by carbon tetrachloride, ethanol, high lipid and assessed biochemically and histologically. Liver function and hydroxyproline contents of liver tissue were determined. Serum hyaluronic acid (HA) level and procollagen III level were performed by radioimmunoassay. The VG staining was used to evaluate the collagen deposit in the liver. Immunohistochemical SABC methods were used to detect transplanted BMSCs and expression of urokinase plasminogen activator (uPA). RESULTS: Serum transaminase level and liver fibrosis in rats were markedly reduced by Guiyuanfang and BMSCs. HA level and procollagen III level were also reduced obviously, compared to model rats (HA: 47.18+/-10.97 ng/mL, 48.96+/-14.79 ng/mL; PCIII: 22.48+/-5.46 ng/mL, 26.90+/-3.35 ng/mL; P<0.05). Hydroxyproline contents of liver tissue in both BMSCs group and Guiyuanfang group were far lower than that of model group (1 227.2+/-43.1 microg/g liver tissue, 1390.8+/-156.3 microg/g liver tissue; P<0.01). After treatment fibrosis scores were also reduced. Both Guiyuanfang and BMSCs could increase the expression of uPA. The transplanted BMSCs could engraft, survive, and proliferate in the liver. CONCLUSION: Guiyuanfang protects against liver fibrosis. Transplanted BMSCs may engraft, survive, and proliferate in the fibrosis livers indefinitely. Guiyuanfang may synergize with BMSCs to improve recovery from liver fibrosis. PMID- 15754397 TI - Effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation. AB - AIM: To study effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation. METHODS: Two of Wistar rats were chosen randomly for normal liver pathology control and ten of SD rats chosen randomly for liver function control as blank group (no operation). The rest of Wistar and SD rats were divided into four groups: control group (only liver transplantation), Dex group (donors receiving intraperitoneal injection of dexamethasone), SpC group (recipients receiving infusion of spleen cells of donors), Dex-SpC group (recipients receiving infusion of apoptotic spleen cells of donors), with each group except blank group, containing 10 SD rats and 10 Wistar rats, respectively. Wistar rats received liver transplantation from SD rats, in the meantime they received infusion of spleen cells of donors, which were induced by an intraperitoneal injection of dexamethasone (3 mg/(d.kg).b.w) for three days before liver transplantation. The serum alanine transaminase (ALT), total bilirubin (T bili), liver pathological changes and survival time were analysed. Statistical analysis was carried out using SPSS 10.0 for Windows. Differences of the parametric data of ALT in means were examined by one-way ANOVA. Differences of ALT between two groups were examined by LSD. Differences of the nonparametric data of T bili in means and scores of pathology classification for acute rejection were examined by Kruskal-Willis H test. The correlations between ALT and T bili were analysed by Bivariate. Kaplan-Meier curves were used to demonstrate survival distribution. The log-rank test was used to compare the survival data. RESULTS: There were significant differences in ALT of the five groups (F = 23.164 P = 0.000), and ALT in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups (P = 0.000), and ALT in SpC group was significantly higher than that in blank control (P = 0.000), control (P = 0.004), and Dex groups (P = 0.02). Results of nonparametric analysis of T bili showed that there were differences in T bili of the five groups (chi(2) = 33.265 P = 0.000). T bili in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups. T bili in SpC group was higher than that in blank control, control, and Dex groups. There were significant differences in scores of pathology classification for acute rejection in each of the groups (chi(2) = 25.933,P = 0.000). The pathologically more serious acute rejection was found in Dex-SPC group than in other groups. No sign of acute rejection was observed in the blank control group. Slight acute rejection was observed in the control group. Slight-moderate acute rejection was observed in the Dex group. Moderate-acute rejection was observed in the SpC group. Severe acute rejection was observed in the Dex-SpC group. The survival time in Dex-SpC group was shorter than in other groups (statistic = 11.13, P = 0.011). ALT and T bili were positively correlated (r = 0.747, P = 0.000, two-tailed). CONCLUSION: In order to reduce quantity of blood loss from rats after liver transplantation, only one of ALT or T bili is needed for liver function measurement of rats. Simultaneous injection of apoptotic spleen cells from donors induced by dexamethasone to liver transplantation rats aggravates acute rejection. One important mechanism of aggravation of acute rejection may be that apoptotic cells are not removed in time and that dead cells including apoptotic cells release inflammatory factors. PMID- 15754398 TI - Clinical significance of serum levels of vascular endothelial growth factor and its receptor in biliary disease and carcinoma. AB - AIM: To investigate the clinical significance of serum vascular endothelial growth factor (VEGF) and soluble VEGF receptor-1 (VEGFR1/Flt-1) (sVEGFR1) levels in biliary diseases. METHODS: We analyzed the serum levels of these proteins in patients with acute cholangitis (group 1), biliary malignancies (group 2), and primary biliary cirrhosis or primary sclerosing cholangitis (group 3), and in healthy donors (group 4). The influence of inflammation was also analyzed. Serum VEGF levels were expressed as VEGF per platelet (VEGF/PLT, pg/10(6)) in order to exclude the influence of platelet counts. RESULTS: sVEGFR1 levels were significantly higher in groups 1 and 2 than in the control group, but did not correlate with inflammatory markers. VEGF/PLT levels were generally higher in patients with active inflammation than in those with carcinoma. C-reactive protein strongly correlated with the levels of serum VEGF independently of platelet and leukocyte counts, even in cancer patients. In cancer patients, VEGF/PLT and sVEGFR1 levels might be indicators for evaluating the effect of medical treatment or the disease progression. CONCLUSION: Serum VEGF and VEGFR1 might be useful markers for gauging the clinical effect of various treatments on patients. PMID- 15754400 TI - An anomaly in persistent right umbilical vein of portal vein diagnosed by ultrasonography. AB - AIM: To detect the anomaly in the persistent right umbilical vein (PRUV) of portal vein (PV) with deviation of the ligamentum tere and left-sided gallbladder. METHODS: A total of 5783 candidates for routine analysis were evaluated for hepatic vascular abnormalities by ultrasonography. RESULTS: Ten candidates (0.17%) had a portal vein anomaly with a rightward-deviated ligamentum tere. The blood-flow velocity in the PRUV of the portal vein (17.7+/-3.0 cm/s) of the 10 cases was similar to that of the right anterior portal trunk (17.6+/-4.1 cm/s). However, the vessel diameter of the PRUV (eta12.4+/-4.4 mm) was larger than the right anterior portal trunk (eta6.1+/-0.9 mm). Therefore, flow volume in the anomalous portion (0.97+/-0.30 L/min) was more than that in the right anterior portal trunk (0.18+/-0.05 L/min). CONCLUSION: The anomaly plays an important role in intra-hepatic PV flow. PMID- 15754399 TI - Synchronous electrogastrographic and manometric study of the stomach as an esophageal substitute. AB - AIM: To investigate the electric and contractile mechanisms involved in the deranged function of the transposed stomach in relation to the course of the symptoms and the changes in contractile and electrical parameters over time. METHODS: Twenty-one patients after subtotal esophagectomy and 18 healthy volunteers were studied. Complaints were compiled by using a questionnaire, and a symptom score was formed. Synchronous electrogastrography and gastric manometry were performed in the fasting state and postprandially. RESULTS: Eight of the operated patients were symptom-free and 13 had symptoms. The durations of the postoperative periods for the symptomatic (9.1+/-6.5 mo) and the asymptomatic (28.3+/-8.8 mo) patients were significantly different. The symptom score correlated negatively with the time that had elapsed since the operation. The percentages of the dominant frequency in the normogastric, bradygastric and tachygastric ranges differed significantly between the controls and the patients. A significant difference was detected between the power ratio of the controls and that of the patients. The occurrence of tachygastria in the symptomatic and the symptom-free patients correlated negatively both with the time that had elapsed and with the symptom score. There was a significant increase in motility index after feeding in the controls, but not in the patients. The contractile activity of the stomach increased both in the controls and in the symptom-free patients. In contrast, in the group of symptomatic patients, the contractile activity decreased postprandially as compared with the fasting state. CONCLUSION: The patients' post-operative complaints and symptoms change during the post-operative period and correlate with the parameters of the myoelectric and contractile activities of the stomach. Tachygastria seems to be the major pathogenetic factor involved in the contractile dysfunction. PMID- 15754401 TI - Long-term follow-up after complete ablation of Barrett's esophagus with argon plasma coagulation. AB - AIM: To report the long-term outcome of patients after complete ablation of non neoplastic Barrett's esophagus (BE) with respect to BE relapse and development of intraepithelial neoplasia or esophageal adenocarcinoma. METHODS: In 70 patients with histologically proven non-neoplastic BE, complete BE ablation was achieved by argon plasma coagulation (APC) and high-dose proton pump inhibitor therapy (120 mg omeprazole daily). Sixty-six patients (94.4%) underwent further surveillance endoscopy. At each surveillance endoscopy four-quadrant biopsies were taken from the neo-squamous epithelium at 2 cm intervals depending on the pre-treatment length of BE mucosa beginning at the neo-Z-line, and from any endoscopically suspicious lesion. RESULTS: The median follow-up of 66 patients was 51 mo (range 9-85 mo) giving a total of 280.5 patient years. A mean of 6 biopsies were taken during surveillance endoscopies. In 13 patients (19.7%) tongues or islands suspicious for BE were found during endoscopy. In 8 of these patients (12.1%) non-neoplastic BE relapse was confirmed histologically giving a histological relapse rate of 3% per year. In none of the patients, intraepithelial neoplasia nor an esophageal adenocarcinoma was detected. Logistic regression analysis identified endoscopic detection of islands or tongues as the only positive predictor of BE relapse (P = 0.0004). CONCLUSION: The long-term relapse rate of non-neoplastic BE following complete ablation with high-power APC is low (3% per year). PMID- 15754403 TI - Transforming growth factor beta can be a parameter of aggressiveness of pT1 colorectal cancer. AB - AIM: To evaluate the significance of transforming growth factor beta (TGF beta) expression, in correlation with histopathological parameters, at the front of invasion in T1 colorectal cancer (CRC) and presence of metastases. METHODS: TGF beta immunohistochemical expression was studied in 34 specimens of colorectal adenocarcinomas (pT1). A three-step avidin-biotinylated immuno-peroxidase (ABCu NCL) staining technique was performed on 4-mum paraffin-embedded tissue sections with a monoclonal antibody to TGF beta (Novocastra, NCL-TGFB, clone TGFB 17, dilution 1:40). RESULTS: Seventeen (50%) out of 34 lesions were positive for TGF beta expression. The TGF beta-positive rate in patients with vascular invasion was significantly higher than in those without vascular invasion (11/14 cases, P<0.01, P = 0.005). The TGF beta-positive rate was observed in 91.7% of patients with presence of tumor budding at the front of invasion (11/12 cases, P<0.01, P = 0.0003). A statistically significant correlation was found between the presence of lymph node metastases and positive expression of TGF beta (14/16 cases, P<0.01, P = 0.0001). We also observed that the TGF beta-positive rates in groups with distant and non-distant metastases were 92.8% and 20% respectively, and a significant correlation between TGF beta expression and distant metastasis was shown (P<0.01, P = 0.00003). CONCLUSION: The evaluation of TGF beta expression of protein in association with histological parameters can be used as a parameter of the aggressiveness of pT1 CRC. PMID- 15754402 TI - IBD5 polymorphisms in inflammatory bowel disease: association with response to infliximab. AB - AIM: Inflammatory bowel diseases (IBD) are multifactorial pathologies of unknown etiology. One susceptibility locus, IBD5, has been mapped to chromosome 5q31. We analyzed our Spanish cohorts of Crohn's disease (CD) and ulcerative colitis (UC) patients to determine whether this locus is associated with IBD, and to ascertain the main clinical phenotype influenced by this risk factor. The kind of interaction, either genetic heterogeneity or epistasis, between this IBD5 susceptibility region and the NOD2/CARD15 gene mutations was studied as well. Finally, we assessed whether this locus can predict response to infliximab therapy. METHODS: A case control study was performed with 274 CD and 211 UC patients recruited from a single center and 511 healthy ethnically matched controls. Two polymorphisms were genotyped in the IBD5 locus and three in the CARD15/NOD2 gene. RESULTS: Our results evidence association only with CD especially with the fistulizing phenotype and in the absence of NOD2/CARD15 variants (mutant allele frequency in patients vs controls: OR = 2.03, 95% CI = 1.35-3.06, P<0.01). The frequency of the IBD5 homozygous mutant genotype significantly increased in CD patients lacking response to infliximab (RR = 3.88, 95% CI = 1.18-12.0, P<0.05). UC patients overall do not show association with 5q31 polymorphisms, although a similar trend to the one observed in CD is found within the worse prognosis group. CONCLUSION: The IBD5 variants may enhance an individual carrier's risk for CD, mainly in the absence of the NOD2/CARD15 mutations and in fistulizing patients. The data presented suggest the potential role of the 5q31 polymorphisms as markers of response to infliximab. PMID- 15754404 TI - Role of the intracellular receptor domain of gp130 (exon 17) in human inflammatory bowel disease. AB - AIM: To study the role of the intracellular receptor domain of gp130 in human inflammatory bowel disease (IBD). METHODS: We amplified and sequenced the complete exon 17 of the human gp130 gene in 146 patients with IBD. According to clinical and histopathological signs, the 146 patients with IBD were classified as having Crohn's disease (n = 73) or ulcerative colitis (n = 63), or as indeterminate status (n = 10). RESULTS: No mutations in exon 17 of the gp130 gene could be detected in any of the 146 patients with IBD examined. CONCLUSION: There is no evidence that mutations in exon 17 of the gp130 gene are involved in the pathogenesis of human IBD. PMID- 15754405 TI - Human papillomavirus in squamous cell carcinoma of esophagus in a high-risk population. AB - AIM: To investigate the relation of human papillomavirus (HPV) and esophageal squamous cell carcinoma (ESCC) in Iranian patients as compared to normal controls. METHODS: Using MY09/MY11 consensus primers, we compared the prevalence of a HPV L1 gene in tumor tissues from 38 ESCC cases and biopsied tissues from 38 endoscopically normal Iranian individuals. We also compared the presence of HPV16 and HPV18 in the same samples using type-specific E6/E7 primers. RESULTS: Fourteen (36.8%) of the 38 ESCC samples but only 5 (13.2%) of the 38 control samples were positive for the HPV L1 gene (P = 0.02). Five (13.2%) of the ESCC samples but none of the control samples were positive for the HPV16 E6/E7 gene (P = 0.05). Three (7.9%) of the ESCC samples and 5 (13.2%) of the control samples were positive for the HPV18 E6/E7 gene (P = 0.71). CONCLUSION: Our data are consistent with HPV DNA studies conducted in other high-risk areas for ESCC. HPV should be considered as a potential factor contributing to the high incidence of ESCC in Iran and other high-incidence areas of the world. PMID- 15754406 TI - Protective effect of Weikang decoction and partial ingredients on model rat with gastric mucosa ulcer. AB - AIM: To investigate the protective mechanisms of Weikang (WK) decoction on gastric mucosae. METHODS: Ninety rats were randomly divided into nine groups of 10 each, namely group, model group, group with large WK dosage, group with medium WK dosage, group with small WK dosage, group with herbs of jianpiyiqi (strengthening the spleen and replenishing qi), group with herbs of yangxuehuoxue (invigorating the circulation of and nourishing the blood), group with herbs of qingrejiedu (clearing away the heat-evils and toxic materials), group with colloidal bismuth pectin (CBP) capsules. According to the method adopted by Yang Xuesong, except normal control group, chronic gastric ulcer was induced with 100% acetic acid. On the sixth day after moldmaking, WK decoction was administered, respectively at doses of 20, 10 and 5 g/kg to rats of the WK groups, or the groups with herbs of jianpiyiqi, yangxuehuoxue and qingrejiedu, 10 mL/kg was separately administered to each group every day. For the group with CBP capsules, medicine was dissolved with water and doses 15 times of human therapeutic dose were administered (10 mL/kg solution containing 0.35% CBP). Rats of other groups were fed with physiological saline (10 mL/kg every day). Administration lasted for 16 d. Rats were killed on d 22 after mold making to observe changes of gastric mucosa. The mucus thickness of gastric mucosa surface was measured. Levels of epidermal growth factor (EGF) in gastric juice, nitric oxide (NO) in gastric tissue, endothelin (ET) in plasma, superoxide dismutase (SOD) in plasma, malondialdehyde (MDA) in plasma and prostaglandin I(2) (PGI(2)) were examined. RESULTS: Compared with control group, ulceration was found in gastric mucosa of model group rats. The mucus thickness of gastric mucosa surface, the levels of EGF, NO, 6-K-PGF(1)alpha and SOD decreased significantly in the model group (EGF: 0.818+/-0.18 vs 2.168+/-0.375, NO: 0.213+/-0.049 vs 0.601+/-0.081, 6-K PGF(1)alpha: 59.7+/-6.3 vs 96.6+/-8.30, SOD: 128.6+/-15.0 vs 196.6+/-35.3, P<0.01), the levels of ET (179.96+/-37.40 vs 46.64+/-21.20, P<0.01) and MDA (48.2+/-4.5 vs 15.7+/-4.8, P<0.01) increased. Compared with model group, the thickness of regenerative mucosa increased, glandular arrangement was in order, and cystic dilative glands decreased, while the mucus thickness of gastric mucosa surface increased (20 g/kg WK: 51.3+/-2.9 vs 23.2+/-8.4, 10 g/kg WK: 43.3+/-2.9 vs 23.2+/-8.4, 5 g/kg WK: 36.1+/-7.2 vs 23.2+/-8.4, jianpiyiqi: 35.4+/-5.6 vs 23.2+/-8.4, yangxuehuoxue: 33.1+/-8.9 vs 23.2+/-8.4, qingrejiedu: 31.0+/-8.0 vs 23.2+/-8.4 and CBP: 38.2+/-3.5 vs 23.2+/-8.4, P<0.05-0.01). The levels of EGF (20 g/kg WK: 1.364+/-0.12 vs 0.818+/-0.18, 10 g/kg WK: 1.359+/-0.24 vs 0.818+/-0.18, 5 g/kg WK: 1.245+/-0.31 vs 0.818+/-0.18, jianpiyiqi: 1.025+/- 0.45 vs 0.818+/ 0.18, yangxuehuoxue: 1.03+/-0.29 vs 0.818+/-0.18, qingrejiedu: 1.02+/-0.47 vs 0.818+/-0.18 and CBP: 1.237+/-0.20 vs 0.818+/-0.18, P<0.05-0.01), NO (20 g/kg WK: 0.480+/-0.026 vs 0.213+/-0.049, 10 g/kg WK: 0.390+/-0.055 vs 0.213+/-0.049, 5 g/kg WK: 0.394+/-0.026 vs 0.213+/-0.049, jianpiyiqi: 0.393+/-0.123 vs 0.213+/ 0.049, yangxuehuoxue: 0.463+/-0.077 vs 0.213+/-0.049, qingrejiedu: 0.382+/-0.082 vs 0.213+/-0.049 and CBP: 0.395+/-0.053 vs 0.213+/-0.049, P<0.05-0.01), 6-K PGF(1)alpha (20 g/kg WK: 86.8+/-7.6 vs 59.7+/-6.3, 10 g/kg WK: 77.9+/-7.0 vs 59.7+/-6.3, 5 g/kg WK: 70.0+/-5.4 vs 59.7+/-6.3, jianpiyiqi: 73.5+/-12.2 vs 59.7+/-6.3, yangxuehuoxue: 65.1+/-5.3 vs 59.7+/-6.3, qingrejiedu: 76.9+/-14.6 vs 59.7+/-6.3, and CBP: 93.7+/-10.7 vs 59.7+/-6.3, P<0.05-0.01) and SOD (20 g/kg WK: 186.4+/-19.9 vs 128.6+/-15.0, 10 g/kg WK: 168.2+/-21.7 vs 128.6+/-15.0, 5 g/kg WK: 155.6+/-21.6 vs 128.6+/-15.0, jianpiyiqi: 168.0+/-85.3 vs 128.6+/-15.0, yangxuehuoxue: 165.0+/-34.0 vs 128.6+/-15.0, qingrejiedu: 168.2+/-24.9 vs 128.6+/ 15.0, and CBP: 156.3+/-18.1 vs 128.6+/-15.0, P<0.05-0.01) significantly increased. The levels of ET (20 g/kg WK: 81.30+/-17.20 vs 179.96+/-37.40, 10 g/kg WK: 83.40+/-25.90 vs 179.96+/-37.40, 5 g/kg WK: 93.87+/-20.70 vs 179.96+/-37.40, jianpiyiqi: 130.67+/-43.66 vs 179.96+/-37.40, yangxuehuoxue: 115.88+/-34.09 vs 179.96+/-37.40, qingrejiedu: 108.22+/-36.97 vs 179.96+/-37.40, and CBP: 91.96+/ 19.0 vs 179.96+/-37.40, P<0.01) and MDA (20 g/kg WK: 21.6+/-7.4 vs 48.2+/-4.5, 10 g/kg WK: 32.2+/-7.3 vs 48.2+/-4.5, 5 g/kg WK: 34.2+/-6.2 vs 48.2+/-4.5, jianpiyiqi: 34.9+/-13.8 vs 48.2+/-4.5, yangxuehuoxue: 35.5+/-16.7 vs 48.2+/-4.5, qingrejiedu: 42.2+/-17.6 vs 48.2+/-4.5, and CBP: 30.1+/-6.1 vs 48.2+/-4.5, P<0.05 0.01) obviously decreased. The 20 g/kg WK group was better than 10 g/kg (the mucus thickness: 51.3+/-2.9 vs 43.3+/-2.9, NO: 0.480+/-0.026 vs 0.390+/-0.055, SOD: 186.4+/-19.9 vs 168.2+/-21.7, P<0.01) and 5 g/kg (the mucus thickness: 51.3+/-2.9 vs 36.1+/-7.2, NO: 0.480+/-0.026 vs 0.394+/-0.026, SOD: 186.4+/-19.9 vs 155.6+/-21.6, P<0.01) groups and CBP group (the mucus thickness: 51.3+/-2.9 vs 38.2+/-3.5, NO: 0.480+/-0.026 vs 0.395+/-0.053, SOD: 186.4+/-19.9 vs 156.3+/ 18.1, P<0.01) in the mucus thickness, NO and SOD levels and better than 10 g/kg (86.8+/-7.6 vs 77.9+/-7.0, P<0.05) and 5 g/kg (86.8+/-7.6 vs 70.0+/-5.4, P<0.05) groups in 6-K-PGF(1)alpha level, 10 g/kg WK group was better than 5 g/kg WK (the mucus thickness: 43.3+/-2.9 vs 36.1+/-7.2, P<0.01, SOD: 168.2+/-21.7 vs 155.6+/ 21.6, P<0.05) and CBP groups (the mucus thickness: 43.3+/-2.9 vs 38.2+/-3.5, P<0.01, SOD: 168.2+/-21.7 vs 156.3+/-18.1, P<0.05) in the mucus thickness and SOD level. In compound group, jianpiyiqi group, yangxuehuoxue group, qingrejiedu group, the level of ET was decreased, NO contents were increased in gastric tissue of ulcers in rats. CONCLUSION: WK decoction and separated recipes have significantly protective effect on ethanol-induced gastric mucosal injury. They can increase the content of EGF in gastric juice, PGI(2) SOD in plasma and NO in gastric tissues, thicken the mucus on the gastric mucosa, and decrease the impairing factor MDA, ET in plasma. PMID- 15754408 TI - Efficacy of different treatment strategies for hepatocellular carcinoma with portal vein tumor thrombosis. AB - AIM: To evaluate the efficacy of different treatment strategies for hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) and investigate factors influencing prognosis. METHODS: One hundred and seventy-nine HCC patients with macroscopic PVTT were enrolled in this study. They were divided into four groups and underwent different treatments: conservative treatment group (n = 18), chemotherapy group (n = 53), surgical resection group (n = 24) and surgical resection with postoperative chemotherapy group (n = 84). Survival rates of the patients were analyzed by the Kaplan-Meier method. A log-rank analysis was performed to identify group differences. Cox's proportional hazards model was used to analyze variables associated with survival. RESULTS: The mean survival periods of the patients in four groups were 3.6, 7.3, 10.1, and 15.1 mo respectively. There were significant differences in the survival rates among the groups. The survival rates at 0.5-, 1-, 2-, and 3-year in surgical resection with postoperative chemotherapy group were 55.8%, 39.3%, 30.4%, and 15.6% respectively, which were significantly higher than those of other groups (P<0.001). Multivariate analysis revealed that the strategy of treatment (P<0.001) and the number of chemotherapy cycles (P = 0.012) were independent survival predictors for patients with HCC and PVTT. CONCLUSION: Surgical resection of HCC and PVTT combined with postoperative chemotherapy or chemoembolization is the most effective therapeutic strategy for the patients who can tolerate operation. Multiple chemotherapeutic courses should be given postoperatively to the patients with good hepatic function reserve. PMID- 15754407 TI - Prognostic significance of cell infiltrations of immunosurveillance in colorectal cancer. AB - AIM: To determine whether the mast cell (MCs) and tumor-associated macrophage (TAMs) counts have any correlation with clinical outcome in colorectal cancer, and to investigate whether MCs undergo phenotypic changes in colorectal cancer. METHODS: The MC and TAM counts were determined immunohistochemically in 60 patients with colorectal cancer and the depth of invasion, lymph node metastasis rate, distant metastasis rates, and survival rates were compared between patients with low (less than the mean number of positive cells) and high (more than the mean number of positive cells) cell counts. RESULTS: Both patients with a low MC count and patients with a low TAM count had significantly deeper depth of invasion than those with a high MC count and those with a high TAM count (P<0.01 and P<0.01 respectively). Patients with a high MC count and patients with a high TAM count were significantly higher showing significantly lower rates of lymph node metastasis, distant metastasis than those with a low MC count and those with a low TAM count. There were significant positive correlation between MC counts and TAM counts (r = 0.852, P<0.01). In both cancerous tissue and normal colorectal tissue, the predominant MC phenotype was MC(TC). The 5-year survival rate estimated was significantly lower in both patients with a low MC count and patients with a low TAM count than in those with a high MC count and those with a high TAM count (P<0.05 and P<0.01 respectively). CONCLUSION: There appears to be a direct relationship between the number of MCs and clinical outcome in patients with colorectal cancer, even though MCs exhibited no significant phenotypic changes. TAM count is of value to predict the clinical outcome or prognosis. It is more beneficial for estimating biological character of colorectal carcinoma to combine MC and TAM counts. PMID- 15754410 TI - Effect of fermented soy milk on the intestinal bacterial ecosystem. AB - AIM: To investigate the effect of fermented soy milk on human ecosystem in the intestinal tract by way of examining the population of different microorganisms isolated from fecal samples. METHODS: A crossover experimental design was applied. Twenty-eight healthy adults completed this experiment. Each subject consumed 250 mL, twice a day between meals, of either fermented soy milk or regular soy milk first for 2 wk, then switched to the other drink after 2 wk. Fecal samples were collected from all subjects every week starting from the second week to the end of the experiment. The microorganisms analyzed were Bifidobacterium spp., Lactobacillus spp., Clostridium perfringens, coliform organisms, and total anaerobic organisms. RESULTS: In the period of fermented soy milk consumption, the populations of Bifidobacterium spp. and Lactobacillus spp. increased (P<0.05) as well as the ratios of Bifidobacterium spp. and Lactobacillus spp. to Clostridium perfringens (P<0.05). The population of coliform organisms decreased (P<0.05) when subjects were in the period of fermented soy milk consumption. CONCLUSION: Intake of fermented soy milk significantly improved the ecosystem of the intestinal tract in the body by increasing the amount of probiotics. PMID- 15754409 TI - CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by the therapy with 5-fluorouracil in murine hepatoma. AB - AIM: To investigate the effect of CpG-containing oligodeoxynucleotides (CpG ODN) alone or in combination with the chemotherapeutic agent 5-fluorouracil (5-FU) on tumor growth and whether CpG ODN can reverse the immunosuppression caused by the chemotherapy with 5-FU in murine hepatoma model. METHODS: Hepatoma model was established by subcutaneous inoculation with hepatoma-22 (H(22)) cells into the right flank of BALB/c mice. Mice with tumor were treated by peritumoral injection of CpG ODN alone or in combination with subcutaneous injection of 5-FU. Tumor size was quantified regularly. Serum levels of IL-12 and IFN-gamma in mice were assayed by enzyme-linked immunosorbent assay (ELISA). The lytic capacity of splenic NK cells was tested by lactate dehydrogenase release assay. RESULTS: Peritumoral injection of CpG ODN alone or in combination with subcutaneous injection of 5-FU, and the treatment with 5-FU alone all led to significant inhibition of hepatoma growth. The mean tumor volumes fell by 46.66% in mice injected with CpG ODN, 68.34% in the 5-FU treated mice, and 70.23% in mice treated with the combination of CpG ODN and 5-FU than in controls. There was no significant difference in tumor size between 5-FU-treated mice and mice treated with the combination of 5-FU and CpG ODN (P>0.05). The serum levels of IL-12 and IFN-gamma of mice treated with CpG ODN alone (IL-12: 464.50+/-24.37 pg/mL; IFN gamma: 134.20+/-25.76 pg/mL) or with the co-administration of CpG ODN and 5-FU (IL-12: 335.83+/-28.74 pg/mL; IFN-gamma: 111.00+/-5.33 pg/mL) were significantly higher than that of controls (IL-12: 237.50+/-45.31 pg/mL; IFN-gamma: 56.75+/ 8.22 pg/mL). The production of IL-12 and IFN-gamma was suppressed moderately in 5 FU-treated mice (IL-12: 166.67+/-53.22 pg/mL; 53.33+/-16.98 pg/mL) compared to control mice (P>0.05), whereas the combination of CpG ODN and 5-FU significantly increased the serum levels of IL-12 and IFN-gamma compared to 5-FU alone (P<0.05). The NK cell killing activity in CpG ODN-treated mice (44.04+/-1.38%) or the mice treated with CpG ODN combined with 5-FU (30.67+/-1.28%) was significantly potentiated compared to controls (19.22+/-0.95%, P<0.05). The co administration of CpG ODN and 5-FU also significantly enhanced the lytic activity of NK cells when compared with the treatment with 5-FU alone (12.03+/-1.42%, P<0.05). CONCLUSION: The present data suggests that CpG ODN used as single therapeutic agent triggers anti-tumor immune response to inhibit the growth of implanted hepatoma and reverses the immunosuppression caused by the chemotherapy with 5-FU. PMID- 15754411 TI - Relationship between expression and distribution of cyclooxygenase-2 and bcl-2 in human gastric adenocarcinoma. AB - AIM: To explore expression and distribution features of COX-2 and bcl-2 in human gastric adenocarcinoma tissues and to study its biological significance. METHODS: Totally 36 human gastric carcinoma samples were enrolled in this study (cardiac adenocarcinoma 16 cases, distal gastric adenocarcinoma 20 cases). The expressions of COX-2 and bcl-2 in cancerous tissues and corresponding para-cancerous tissues were investigated by immunohistochemistry using COX-2 polyclonal antibody and bcl 2 monoclonal antibody. The normal gastric mucosa tissues were used as control. RESULTS: The expressions of COX-2 and bcl-2 in gastric carcinoma were significantly higher than that in the para-cancerous tissues (77.8% vs 47.2%, P<0.01, 80.56% vs 58.33%, P<0.05). The expression of COX-2 in cardiac adenocarcinoma was remarkably higher than that in the distal gastric carcinoma (93.8% vs 65.0%, P<0.01). The expression of COX-2 was mainly localized in the cytoplasm of tumor cells and partly in the nucleus. There is a transition of the COX-2 cytoplasmic positivity to nucleic in tumor cells with the increase of gastric carcinoma pathological grade. Interstitial macrophages, fibroblasts and vascular endothelial cells also expressed COX-2. The tissues with higher expression of COX-2 also expressed high level of bcl-2 protein. CONCLUSION: Abnormal expression pattern of COX-2 within the tissues of human gastric cancer is correlated with tumor location and lymph node metastasis. COX-2 may regulate expression of apoptosis suppressor gene (bcl-2) through interaction of tumor cells and stromal cells and play an important role in the generation and development of tumors, which will be of great help in developing new methods for antitumor therapy. PMID- 15754412 TI - Expression of TNF-alpha and VEGF in the esophagus of portal hypertensive rats. AB - AIM: To investigate the expression of tumor necrosis factor-alpha (TNF-alpha) and vascular endothelial growth factor (VEGF) in the development of esophageal varices in portal hypertensive rats. METHODS: Thirty male Sprague-Dawley (SD) rats in the model group in which a two-stage ligation of portal vein plus ligation of the left adrenal vein was performed, were divided into three subgroups (M(7), M(14), and M(21)) in which the rats were kiued on the seventh day, the 14(th) d and the 21 d after the complete portal ligation. Thirty male SD rats, which underwent the sham operation in the control group, were also separated into three subgroups (C(7), C(14) and C(21)) corresponding to the models. The expression of TNF-alpha and VEGF in the esophagus of all the six subgroups of rats were measured with immunohistochemical SP technique. RESULTS: The portal pressure in the three model subgroups was significantly higher than that in the corresponding control subgroups (23.82+/-1.83 vs 11.61+/-0.86 cmH(2)O, 20.90+/-3.27 vs 11.43+/-1.55 cmH(2)O and 20.68+/-2.27 vs 11.87+/-0.79 cmH(2)O respectively, P<0.01), as well as the number (9.3+/-1.6 vs 5.1+/-0.8, 11.1+/-0.8 vs 5.4+/-1.3 and 11.7+/-1.5 vs 5.2+/-1.1 respectively, P<0.01) and the total vascular area (78 972.6+/-3 527.8 vs 12 993.5+/-4 994.8 mum(2), 107 207.5+/ 4 6461.4 vs 11 862.6+/-5 423.2 mum(2) and 110 241.4+/-49 262.2 vs 11 973.7+/-3 968.5 mum(2) respectively, P<0.01) of submucosal veins in esophagus. Compared to the corresponding controls, the expression of TNF-alpha and VEGF in M(21) was significantly higher (2.23+/-0.30 vs 1.13+/-0.28 and 1.65+/-0.38 vs 0.56+/-0.30 for TNF-alpha and VEGF respectively, P<0.01), whereas there was no difference in M(7) (1.14+/-0.38 vs 1.06+/-0.27 and 0.67+/-0.35 vs 0.50+/-0.24 for TNF-alpha and VEGF respectively, P>0.05) and M(14) (1.20+/-0.25 vs 1.04+/-0.26 and 0.65+/-0.18 vs 0.53+/-0.25 for TNF-alpha and VEGF respectively, P>0.05). And the expression of TNF-alpha and VEGF in M(21) was significantly higher than that in M(7) (2.23+/ 0.30 vs 1.14+/-0.38 and 1.65+/-0.38 vs 0.67+/-0.35 for TNF-alpha and VEGF respectively, P<0.01) and M(14) (2.23+/-0.30 vs 1.20+/-0.25 and 1.65+/-0.38 vs 0.65+/-0.18 for TNF-alpha and VEGF respectively, P<0.01), but there was no difference between M(7) and M(14) (1.14+/-0.38 vs 1.20+/-0.25 and 0.67+/-0.35 vs 0.65+/-0.18 for TNF-alpha and VEGF respectively, P>0.05). CONCLUSION: In the development of esophageal varices in portal hypertensive rats, increased TNF alpha and VEGF may be not an early event, and probably play a role in weakening the esophageal wall and the rupture of esophageal varices. PMID- 15754413 TI - Expression of p57(kip2) and its relationship with clinicopathology, PCNA and p53 in primary hepatocellular carcinoma. AB - AIM: To investigate the expression of p57(kip2) and its relationship with clinicopathology, PCNA and p53 in primary hepatocellular carcinoma (HCC). METHODS: Expression of p57(kip2), PCNA and p53 in tumor tissues from 32 patients with HCC and 10 liver tissues of normal persons was detected with Elivision immunohistochemical technique. RESULTS: The p57(kip2) protein positive-expression rate in HCC was 56.25%, lower than that in normal tissues (100%, P<0.05). The reduced expression of p57(kip2) protein correlated significantly with moderate or low differentiation of tumor cells (P = 0.007 <0.05), high clinical stage (P = 0.041 <0.05) and poor prognosis (P = 0.036 <0.05), but did not correlate significantly with metastasis, tumor size, level of AFP and age (P>0.05). The PCNA positive-expression rate was 56.25%, which was correlated significantly with the expression of p57(kip2) (P = 0.025<0.05). The p53 positive-expression rate was 46.88%, which was not correlated significantly with the expression of p57(kip2) (P>0.05). CONCLUSION: There is a marked loss or absence of p57(kip2) expression and high expression of PCNA in HCC, which are involved in carcinogenesis and development of HCC. The p57(kip2) and p53 may induce apoptosis via different mechanisms. PMID- 15754414 TI - Liver cirrhosis as a consequence of iron overload caused by hereditary nonspherocytic hemolytic anemia. AB - Nonspherocytic hereditary anemias are occasionally accompanied by significant iron overload but the significance for the development of chronic liver disease is not clear. We described two cases of patients with chronic liver disease and severe iron overload due to chronic hereditary hemolysis. Both patients have had signs of liver cirrhosis and severe hemolysis since childhood. A hereditary pyruvate kinase deficiency (PKD) was discovered as the underlying reason for the hemolysis. Sequencing of the pyruvate kinase gene showed a mutation within exon 11. Liver histology in both patients revealed cirrhosis and a severe iron overload but primary hemochromatosis was excluded by HFE-gene analysis. An iron reduction therapy with desferrioxamine led to significant decrease of serum ferritin and sustained clinical improvement. PKD-induced hemolysis may cause severe iron overload even in the absence of HFE-genotype abnormalities. This secondary iron overload can lead to chronic liver disease and cirrhosis. Therefore, the iron metabolism of PKD patients has to be closely monitored and iron overload should be consequently treated. PMID- 15754415 TI - Clostridium difficile causing acute renal failure: case presentation and review. AB - AIM: Clostridium difficile infection is primarily a nosocomial infection but asymptomatic carriers of Clostridium difficile can be found in up to 5% of the general population. Ampicillin, cephalosporins and clindamycin are the antibiotics that are most frequently associated with Clostridium difficile associated diarrhea or colitis. Little is known about acute renal failure as a consequence of Clostridium difficile-associated diarrhea. METHODS: In this case report, we describe the course of Clostridium difficile-associated diarrhea in an 82-year-old patient developing acute renal failure. Stopping the offending agent and symptomatic therapy brought a rapid improvement of diarrhea and acute renal failure, full recovery was gained 18 d after admission. In a systematic review we looked for links between the two conditions. RESULTS: The link between Clostridium difficile-associated diarrhea and acute renal failure in our patient was most likely volume depletion. However, in experimental studies a direct influence of Clostridium difficile toxins on renal duct cells could be shown. CONCLUSION: Rapid diagnosis, nonspecific supportive treatment and specific antibiotic treatment, especially in the elderly, may lower excess mortality Clostridium difficile-associated diarrhea and renal failure being possible complications. PMID- 15754416 TI - Report of gossypiboma from the standpoint in medicine and law. AB - We report on a case of gossypiboma. A 78-year-old man was admitted to our hospital with acute abdomen. He had undergone an operation for colon cancer 4 mo previously. Abdominal ultrasonography revealed an echogenic lesion with a hypoechoic rim and strong posterior acoustic shadowing in the lower abdomen. Diagnosis of gossypiboma can be made by ultrasonography to avoid loss-of-chance of survival. We reviewed the English literature briefly of gossypiboma from the medical and juridical view. According to the theory of loss-of-chance, the damage of plaintiff is the loss of the chance of survival or recovery, rather than the final harm. The victim would allow recovery for the loss of the chance from the defendant. But the plaintiff would show by a preponderance that he was deprived of a better chance of a cure. Under the proposed rule, the compensable value of the victim would be the plaintiff's compensation for the loss of the victim's chance of survival. PMID- 15754417 TI - An integrated approach utilizing proteomics and bioinformatics to detect ovarian cancer. AB - OBJECTIVE: To find new potential biomarkers and establish the patterns for the detection of ovarian cancer. METHODS: Sixty one serum samples including 32 ovarian cancer patients and 29 healthy people were detected by surface-enhanced laser desorption/ionization mass spectrometry (SELDI-MS). The protein fingerprint data were analyzed by bioinformatics tools. Ten folds cross-validation support vector machine (SVM) was used to establish the diagnostic pattern. RESULTS: Five potential biomarkers were found (2085 Da, 5881 Da, 7564 Da, 9422 Da, 6044 Da), combined with which the diagnostic pattern separated the ovarian cancer from the healthy samples with a sensitivity of 96.7%, a specificity of 96.7% and a positive predictive value of 96.7%. CONCLUSIONS: The combination of SELDI with bioinformatics tools could find new biomarkers and establish patterns with high sensitivity and specificity for the detection of ovarian cancer. PMID- 15754418 TI - The effect of hepatic blood inflow occlusion on hepatic cancer treated with diode laser thermocoagulation. AB - OBJECTIVE: To assess the effect of temporary occlusion of hepatic blood inflow on hepatic cancer treated with diode-laser induced thermocogation (LITT). METHODS: The carcinoma Walker-256 was implanted in 40 SD rat livers. Twelve days later, the animals were randomly divided into 4 groups. Group A received LITT alone; group B received hepatic artery temporary occlusion during LITT; group C received portal vein temporary occlusion during LITT; group D received hepatic artery and portal vein temporary occlusion during LITT. Tumors were exposed to 810 nm diode laser light at 0.95 watts for 10 min from a scanner tip applicator placed in the tumor. At the same time, the intrahepatic temperature distribution in rats with liver tumors was measured per 2 min during thermocoagulation. Tumor control was examined immediately 7 and 14 d after thermocoagulation. RESULTS: There was significant difference of intrahepatic temperature distribution in rats with liver tumors among the 4 groups (P<0.05) except when group C samples were compared with group D samples at each time point, and group B samples were compared with group C samples at 120 s (P>0.05). Light microscopic examination of the histologic section samples revealed three separate zones: regular hyperthermic coagulation necrosis zone, transition zone and reference zone. Compared with the samples in group A and group B, group C and group D samples had more clear margin among the three zones. CONCLUSION: The hepatic blood inflow occlusion, especially portal vein hepatic blood inflow occlusion, or all hepatic blood inflow occlusion considerably increased the efficacy of LITT in the treatment of liver cancer. PMID- 15754420 TI - Human bone marrow-derived mesenchymal stem cells transplanted into damaged rabbit heart to improve heart function. AB - OBJECTIVE: The present study was designed to test whether transplantation of human bone marrow-derived mesenchymal stem cells (hMSCs) in New Zealand rabbits with myocardial infarction can improve heart function; and whether engrafted donor cells can survive and transdifferentiated into cardiomyocytes. METHODS: Twenty milliliters bone marrow was obtained from healthy men by bone biopsy. A gradient centrifugation method was used to separate bone marrow cells (BMCs) and red blood cells. BMCs were incubated for 48 h and then washed with phosphate buffered saline (PBS). The culture medium was changed twice a week for 28 d. Finally, hematopoietic cells were washed away to leave only MSCs. Human MSCs (hMSCs) were premarked by BrdU 72 h before the transplantation. Thirty-four New Zealand rabbits were randomly divided into myocardial infarction (MI) control group and cell treated group, which received hMSCs (MI+MSCs) through intramyocardial injection, while the control group received the same volume of PBS. Myocardial infarction was induced by ligation of the left coronary artery. Cell treated rabbits were treated with 5 x 10(6) MSCs transplanted into the infarcted region after ligation of the coronary artery for 1 h, and the control group received the same volume of PBS. Cyclosporin A (oral solution; 10 mg/kg) was provided alone, 24 h before surgery and once a day after MI for 4 weeks. Echocardiography was measured in each group before the surgery and 4 weeks after the surgery to test heart function change. The hearts were harvested for HE staining and immunohistochemical studies after MI and cell transplantation for 4 weeks. RESULTS: Our data showed that cardiac function was significantly improved by hMSC transplantation in rabbit infarcted hearts 4 weeks after MI (ejection fraction: 0.695+/-0.038 in the cell treated group (n=12) versus 0.554+/-0.065 in the control group (n=13) (P<0.05). Surviving hMSCs were identified by BrdU positive spots in infarcted region and transdifferentiated into cardiomyocytes characterized with a positive cardiac phenotype: troponin I. CONCLUSION: Transplantation of hMSCs could transdifferentiate into cardiomyocytes and regenerate vascular structures, contributing to functional improvement. PMID- 15754419 TI - Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression. AB - OBJECTIVES: To evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro. METHODS: (1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry. RESULTS: Our data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The expression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment. CONCLUSION: Our results demonstrated that siRNAs exerted robust and specific inhibition on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency. PMID- 15754421 TI - The effects of sotalol on ventricular repolarization during exercise. AB - OBJECTIVE: Although after pacing animal and human studies have demonstrated a rate-dependent effect of sotalol on ventricular repolarization, there is little information on the effects of sotalol on ventricular repolarization during exercise. This study attempted to show the effects of sotalol on ventricular repolarization during physiological exercise. METHODS: Thirty-one healthy volunteers (18 males, 13 females) were enrolled in the study. Each performed a maximal treadmill exercise test according to the Bruce protocol after random treatment with sotalol, propranolol and placebo. RESULTS: Sotalol significantly prolonged QTc (corrected QT) and JTc (corrected JT) intervals at rest compared with propranolol (QTc 324.86 ms vs 305.21 ms, P<0.001; JTc 245.04 ms vs 224.17 ms, P<0.001) and placebo (QTc 324.86 ms vs 314.06 ms, P<0.01; JTc 245.04 ms vs. 232.69 ms, P<0.001). The JTc percent reduction increased progressively with each stage of exercise and correlated positively with exercise heart rate (r=0.148, P<0.01). The JTc percent reduction correlation with exercise heart rate did not exist with either propranolol or placebo. CONCLUSIONS: These results imply that with sotalol ventricular repolarization is progressively shortened after exercise. Thus the specific class III antiarrhythmic activity of sotalol, present as delay of ventricular repolarization, may be attenuated during exercise. Such findings may imply the need to consider other antiarrythmic therapy during periods of stress-induced tachycardia. PMID- 15754422 TI - GM1 stabilizes expression of NMDA receptor subunit 1 in the ischemic hemisphere of MCAo/reperfusion rat. AB - OBJECTIVE: To determine the protective effect of monosialoganglionside (GM1) and evaluate the influence of GM1 on expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in Sprague-Dawley (SD) rats with focal cerebral ischemia reperfusion (I/R). METHODS: Left middle cerebral artery (MCA) was occluded by an intraluminal suture for 1 h and the brain was reperfused for 72 h in SD rats when infarct volume was measured, GM1 (10 mg/kg) was given ip (intraperitoneally) at 5 min (group A), 1 h (group B) and 2 h (group C) after MCA occlusion (MCAo). Expression of NMDAR1 was detected by Western blot at various time after reperfusion (4 h, 6 h, 24 h, 48 h and 72 h) in ischemic hemispheres of the rats with or without GM1 administered. RESULTS: (1) Adjusted relative infarct volumes of groups A and B were significantly smaller than that of group C and the control group (P<0.01 and P<0.05, respectively). (2) Expression level of NMDAR1 was temporally high at 6 h after reperfusion, and dipped below the normal level at 72 h after reperfusion. GM1 at 5 min after MCAo significantly suppressed the expression of NMDAR1 at 6 h after reperfusion (P<0.05 vs the control). At 72 h after reperfusion, the NMDAR1 expression level of rats treated with GM1 administered (at 5 min or 2 h after MCAo) was significantly higher than that of the control (P<0.05). CONCLUSION: GM1 can time-dependently reduce infarct volume in rats with focal cerebral I/R partly through stabilizing the expression of NMDAR1. PMID- 15754423 TI - Abnormal cortical functional connections in Alzheimer's disease: analysis of inter- and intra-hemispheric EEG coherence. AB - To investigate inter- and intra-hemispheric electroencephalography (EEG) coherence at rest and during photic stimulation of patients with Alzheimer's disease (AD). Thirty-five patients (12 males, 23 females; 52 to approximately 64 y) and 33 sex- and age-matched controls (12 males, 21 females; 56 to approximately 65 y) were recruited in the present study. EEG signals from C3-C4, P3-P4, T5-T6 and O1-O2 electrode pairs resulted from the inter-hemispheric action, and EEG signals from C3-P3, C4-P4, P3-O1, P4-O2, C3-O1, C4-O2, T5-O1 and T6-O2 electrode pairs resulted from the intra-hemispheric action. The influence of inter- and intra-hemispheric coherence on EEG activity with eyes closed was examined, using fast Fourier transformation from the 16 sampled channels. The frequencies of photic stimulation were fixed at 5, 10 and 15 Hz, respectively. The general decrease of AD patients in inter- and intra-hemispheric EEG coherence was more significant than that of the normal controls at the resting EEG, with most striking decrease observed in the alpha-1 (8.0-9.0 Hz) and alpha-2 (9.5-12.5 Hz) bands. During photic stimulation, inter- and intra-hemispheric EEG coherences of the AD patients having lower values in the alpha (9.5-10.5 Hz) band than those of the control group. It suggests that under stimulated and non-stimulated conditions, AD patients had impaired inter- and intra-hemispheric functional connections, indicating failure of brain activation in alpha-related frequency. PMID- 15754425 TI - Clinical detection and movement recognition of neuro signals. AB - Neuro signal has many more advantages than myoelectricity in providing information for prosthesis control, and can be an ideal source for developing new prosthesis. In this work, by implanting intrafascicular electrode clinically in the amputee's upper extremity, collective signals from fascicules of three main nerves (radial nerve, ulnar nerve and medium nerve) were successfully detected with sufficient fidelity and without infection. Initial analysis of features under different actions was performed and movement recognition of detected samples was attempted. Singular value decomposition features (SVD) extracted from wavelet coefficients were used as inputs for neural network classifier to predict amputee's movement intentions. The whole training rate was up to 80.94% and the test rate was 56.87% without over-training. This result gives inspiring prospect that collective signals from fascicules of the three main nerves are feasible sources for controlling prosthesis. Ways for improving accuracy in developing prosthesis controlled by neuro signals are discussed in the end. PMID- 15754424 TI - Inhibitory effects of jujuboside A on EEG and hippocampal glutamate in hyperactive rat. AB - In this study, the inhibitory effect of jujuboside A (JuA) on a penicillin sodium (Na-PCN) induced hyperactivity model was investigated. Cortical EEG (electroencephalogram) and the concentration of hippocampal Glutamate (Glu) were monitored simultaneously in vivo as indicators of rat's excitatory state. Power spectral density (PSD) and gravity frequency of PSD were calculated. JuA (0.05 g/L and 0.1 g/L) inhibited the EEG excitation effect caused by Na-PCN by increasing the power of delta1 and delta2 bands (P<0.01 vs model) and lowering the gravity frequency of PSD (P<0.01 vs model). JuA also remarkably reduced the Glu elevation induced by Na-PCN (P<0.05 vs model). Diazepam also depressed Glu concentration and lowered the gravity frequency, but it showed a different EEG pattern in increased beta2-activity (P<0.01 vs model). EEG excitation caused by Na-PCN correlated with Glu elevation during the first hour. Neurophysiological inhibitory effects of JuA and diazepam were more persistent than their Glu inhibitory effects. PMID- 15754426 TI - Construction of a hepatic stellate cells subtracted cDNA library of differentially expressed genes in normal mice and mice with Schistosomiasis japonica. AB - To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosoma japonicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5alpha. The amplified library contained more than 400 positive bacterial clone, which were then hybridized with forward and backward subtracted probes for differential screening. One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis. PMID- 15754427 TI - Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B. AB - OBJECTIVE: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. METHODS: The total RNA from liver tissue with chronic hepatitis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5' end of the RNA transcript (SMART) technique and CDS III/3' primer were used for first strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to lambdaTriplEx2 vector. Then lambda phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. RESULTS: The titers of unamplifed and amplified libraries were 1.94 x 10(6) pfu/ml and 1.49 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1-2 kb in 64.29%, and 0.5-1.0 kb in 35.71%. CONCLUSION: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed. PMID- 15754428 TI - Proapoptotic and pronecrosis effect of different truncated hepatitis C virus core proteins. AB - OBJECTIVE: To study the roles of different truncated hepatitis C virus (HCV) core proteins (CORE) in the pathogenesis of HCV persistent infection and hepatocellular carcinoma (HCC) and to assess intracellular localization in transiently transfected cells. METHODS: Seven truncated GFP (green fluorescent protein)-CORE fusion protein expression plasmids were constructed, which contained HCV CORE sequences derived from tumor tissues (BT) and non-tumor tissues (BNT) from one patient infected with HCV. Amino acid (aa) lengths were BT: 1-172 aa, 1-126 aa, 1-58 aa, 59-126 aa, 127-172 aa; BNT: 1-172 aa and C191: 1 172 aa respectively. Subcellular localization of CORE-GFP was analyzed by con focal laser scanning microscope. Apoptosis and necrosis were quantified by flow cytometry. RESULTS: Different truncated CORE-GFP localized mainly in the cytoplasm, but nuclear staining was also observed. HCV CORE could induce apoptosis and necrosis, and different truncated COREs could induce cell apoptosis and necrosis at different levels. Among the same length 1-172 aa of BT, BNT and C191, the cell apoptosis and necrosis percentage of BT is highest, and C191 is the lowest (BT>BNT>C191). To the different fragment COREs of BT, N-terminal of CORE induced apoptosis and necrosis higher, compared with that of C-terminal (1 172 aa>1-126 aa>1-58 aa>127-172 aa>59-126 aa). CONCLUSION: These results suggest HCV CORE could induce apoptosis and necrosis of cells, which might play an important role in the pathogenesis of HCV persistent infection and HCC and the different CORE domains of different HCV quasi-species might have some difference in their pathogenesis. PMID- 15754429 TI - Perioperative nursing for laparoscopic liver resection. AB - Sir Run Run Shaw Hospital (SRRSH) has developed a form of laparoscopic hepatectomy, resecting by curettage and suction. Such resection has been carried out successfully in 6 patients who had liver tumors. The results are satisfactory. And after the operation, there is a very effective perioperative nursing ensuring the patient's recovery. PMID- 15754430 TI - Serious response during tilt-table test in elderly and its prophylactic management. AB - OBJECTIVE: To evaluate the serious response during tilt-table test (TTT) and its prophylactic management. METHOD: Seventy-six elderly patients were tested at a tilt angle of 70 degrees for a maximum of 45 min and then subjected to isoproterenol-provocative tilt testing. ECG and blood pressure were monitored during the test and patients were kept at normal saline condition through a peripheral intravenous duct. RESULTS: Fifty-one of 76 patients were defined as positive including 23 having serious response; 6 of the 23 patients had arteriosclerosis involving internal carotid arteries and 7 cases had bradycardia, two of which were associated with II degrees -I A-V block and the others with chronic atrial fibrillation. The serious response consisted of cardiac arrest for more than 5 s (6 cases), or serious bradycardia for more than 1 min (7 cases) or serious hypotension for more than 1 min (10 cases). Those with serious response were managed by returning to supine position, thus driving up legs and intravenous atropine, CPR (2 cases with cardiac arrest) and needing oxygen supplementation (11 cases). Only 2 hypotension patients recovered gradually by 10 min after emergency management, while others recovered rapidly with no complications. CONCLUSION: Although non-invasive, TTT may result in serious response, especially in elderly. Therefore proper patient selection, control of isoproterenol infusion and close observation of vital signs are decisive for a safe consequence. PMID- 15754431 TI - [Illness due to work]. PMID- 15754432 TI - Taking the lead. PMID- 15754433 TI - Angina pectoris presenting as headache. PMID- 15754434 TI - Out-of-hours cover. PMID- 15754435 TI - Out-of-hours cover. PMID- 15754436 TI - Small Animal Medicine Society. PMID- 15754437 TI - [Pulmonary hypertension in chronic obstructive lung disease]. PMID- 15754438 TI - Building a healthy North Dakota position statement on tobacco tax increase. PMID- 15754439 TI - Magnetic resonance imaging in Tolosa-Hunt syndrome. PMID- 15754440 TI - Perspectives. Gearing up for a game of Medicaid hot potato. PMID- 15754441 TI - [Medicinal cannabis for diseases of the nervous system: no convincing evidence of effectiveness]. PMID- 15754442 TI - Radiolabeled J591 antibody delivers lethal hit to advance prostate cancers in a phase I trail. PMID- 15754443 TI - Perturbing and detecting phospho-protein networks in single leukemia cells. PMID- 15754444 TI - Cell cycle abnormalities may be key to aggressive breast cancer in African American women. PMID- 15754445 TI - Overexpressed DLC1 as target for breast cancer cell shape and survival. PMID- 15754446 TI - Coxibs and thrombosis. PMID- 15754447 TI - Prescribed minimum benefits or minimum prescribed benefits? PMID- 15754448 TI - Dose sparing with intradermal injection of influenza vaccine. PMID- 15754449 TI - Recording patient data. PMID- 15754450 TI - Our interprofessional future. PMID- 15754451 TI - Revised guidelines for the treatment of OIs. PMID- 15754453 TI - Avoid PPIs with atazanavir. PMID- 15754452 TI - Efavirenz reclassified as FDA pregnancy category D. PMID- 15754454 TI - Is resistance testing of benefit in treatment-naive HIV-infected patients? PMID- 15754455 TI - Nutrition. How important is glutamine supplementation in HIV infection? PMID- 15754456 TI - Dentistry. Implications and management of xerostomia in the HIV patient. PMID- 15754457 TI - Mental health. Ethics committees can help with HIV-related ethical dilemmas. PMID- 15754458 TI - Advances in the pharmaceutical care of hepatitis C/HIV co-infection. PMID- 15754462 TI - A Festschrift in honor of the 60th birthday of Professor Hiroshi Masuhara. PMID- 15754463 TI - [Surgical treatment of peripheral nerve lesions]. PMID- 15754464 TI - Plasma phospholipid transfer protein activity and subclinical inflammation in type 2 diabetes mellitus. AB - Phospholipid transfer protein (PLTP) transfers phospholipids between lipoproteins, and plays an essential role in HDL metabolism. The regulation of PLTP is poorly understood and recent evidence suggests that PLTP activity increases during acute-phase response. Since type 2 diabetes is associated with chronic subclinical inflammation, the objective is to determine whether inflammation modulates PLTP in diabetes. Plasma PLTP activity was assayed by measuring the transfer of radiolabeled phosphatidylcholine from liposomes to HDL and high-sensitivity C-reactive protein (CRP) by immunoturbidimetric assay in 280 type 2 diabetic patients and 105 controls. Plasma PLTP activity (2364+/-651 nmol/ml/h versus 1880+/-586 nmol/ml/h in control, mean +/- S.D., P <0.01) and CRP (1.64(0.89-3.23)mg/l versus 0.99(0.53-2.23 mg/l, median (interquartile range), P<0.01) were increased in diabetic subjects. PLTP activity correlated significantly with age, BMI, HbA1c, log(CRP) and apolipoprotein AI and B in diabetic subjects. General linear model analysis showed that only apolipoprotein AI, age, BMI, and log(CRP) were independent determinants of PLTP activity. In conclusion, PLTP activity is increased in diabetes and apolipoprotein AI is a major determinant of PLTP activity. There is also an independent association between CRP and PLTP activity, suggesting that subclinical inflammation may influence PLTP activity in diabetes. PMID- 15754465 TI - Cell-mediated protein release from calcium-phosphate-coated titanium implants. PMID- 15754466 TI - Responses of cultured macrophages to microspheres. PMID- 15754467 TI - Medicare program: changes to the Medicare claims appeal procedures. Interim final rule with comment period. AB - Medicare beneficiaries and, under certain circumstances, providers and suppliers of health care services, can appeal adverse determinations regarding claims for benefits under Medicare Part A and Part B under sections 1869 and 1879 of the Social Security Act (the Act). Section 521 of the Medicare, Medicaid, and SCHIP Benefits Act of 2000 (BIPA) amended section 1869 of the Act to provide for significant changes to the Medicare claims appeal procedures. This interim final rule responds to comments on the November 15, 2002 proposed rule regarding changes to these appeal procedures, establishes the implementing regulations, and explains how the new procedures will be implemented. It also sets forth provisions that are needed to implement the new statutory requirements enacted in Title IX of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). PMID- 15754468 TI - Report of meeting on the development of influenza vaccines with broad spectrum and long-lasting immune responses, World Health Organization, Geneva, Switzerland, 26-27 February 2004. PMID- 15754469 TI - Polycystic ovaries. PMID- 15754471 TI - Update on comparison of current prediction imaging programs. PMID- 15754470 TI - Extracellular matrix proteins and the selective resorption of deciduous tooth roots. PMID- 15754473 TI - Archer John Porter Martin CBE 1 March 1910 - 28 July 2002. AB - We judge the worth of a scientist by the benefits he or she brings to science and society; by this measure Archer Martin was outstanding, and rightfully his contribution was recognized with a Nobel Prize. Scientific instruments and instrumental methods now come almost entirely from commercial sources and we take them for granted and often have little idea how they work. Archer Martin was of a different time when scientists would often devise their own new instruments, which usually they fully understood, and then they would use them to explore the world. The chromatographic methods and instruments Martin devised were at least as crucial in the genesis and development of molecular biology as were those from X-ray crystallography. Liquid partition chromatography, especially in its two dimensional paper form, revealed the amino acid composition of proteins and the nucleic acid composition of DNA and RNA with a rapid and elegant facility. Gas chromatography (GC) enabled the accurate and rapid analysis of lips, which previously had been painfully slow and little more than a greasy sticky confusion of beaker chemistry. Martin's instruments enabled progress in the sciences ranging from geophysics to biology , and without im we might have waited decades before another equivalent genius appeared. More than this, the environmental awareness that Rachel Carson gave us would never had solidified as it did without the evidence of global change measured by GC. This instrumental method provided accurate evidence about the ubiquity of pesticides and pollutants and later made us aware of the growing accumulation in the atmosphere of chlorinated fluorocarbons, nitrous oxide and other ozone-depleting chemicals. If all this were not enough to glorify Martin's partition chromatography, there is the undoubted fact that its simplicity, economy and exquisite resolving power transformed the chemical industry and made possible so many of the conveniences we now take for granted. PMID- 15754472 TI - Sir William MacGregor Henderson 17 July 1913 - 29 November 2000. AB - W. M. 'Gregor' Henderson belonged to the great tradition of veterinary involvement in the control of epizootic diseases that was such a feature of the middle part of the twentieth century. He was one of the pioneers of research into the virology of foot-and-mouth disease and the development and application in the field of vaccines to control it. THroughout his career, first as scientists, latterly as administrator, he maintained a close interest in the animals to whose wellbeing research was directed and in the work of the practising veterinary surgeons who ultimately translated veterinary science into veterinary practice. PMID- 15754474 TI - Loading effects on rat craniomandibular morphology: a system for gravity studies. AB - Gravity effects on muscle and bone are a major impediment to long-term space travel. We introduce a model for studying these effects, the craniomandibular system. Some advantages of this system include: (1) craniomandibular morphology is determined by epigenetic factors including gravity, (2) relatively light forces can significantly alter its morphology, and (3) soft diet and tooth loss produce effects that are similar to those produced in lower limbs by weightlessness. In the study, implants made either of gold (experimental group) or lightweight acrylic (controls) were attached to adult rats' mandibles. After 13 weeks, the animals' skulls and mandibles were dissected. Pair-wise comparisons indicated that the experimental animals showed significantly shortened and narrowed cranial bases, and significant changes in the posterior zygomatic arch region. These results indicate that simulated macrogravity influences bone remodeling in the adult craniomandibular system. PMID- 15754475 TI - Effects of HZE particle on the nigrostriatal dopaminergic system in a future Mars mission. AB - Because of long duration travel outside the Earth's magnetic field, the effect of iron-rich high charge and energy (HZE) particles in Galactic Cosmic Rays on human body is the major concern in radiation protection. Recently attention has been directed to effects on the central nervous system in addition to mutagenic effects. In particular, a reduction in striatal dopamine content on nigrostriatal dopaminergic system has been reported by investigators using accelerated iron ions in ground-based mammalian studies. In addition, studies of the pathophysiology of Parkinson's disease demonstrated that excess iron cause a reduction in the dopamine content in the substantia nigra. This suggests an intriguing possibility to explain the selective detrimental effects of HZE particles on the dopaminergic system. Should these particles have biochemical effects, possible options for countermeasures are: (1) nutritional prevention, (2) medication, and (3) surgical placement of a stimulator electrode at a specific anatomic site in the basal ganglia. PMID- 15754476 TI - Vanguard--a European robotic astrobiology-focussed Mars sub-surface mission proposal. AB - We present a new European Mars mission proposal to build on the UK-led Beagle2 Mars mission and continue its astrobiology-focussed investigation of Mars. The small surface element to be delivered to the Martian surface--Vanguard--is designed to be carried by a Mars Express-type spacecraft bus to Mars and adopts a similar entry, descent and landing system as Beagle2. The surface element comprises a triad of robotic devices--a lander, a micro-rover of the Sojourner class for surface mobility, and three ground-penetrating moles mounted onto the rover for sub-surface penetration to 5 m depth. The major onboard instruments on the rover include a Raman spectrometer/imager, a laser plasma spectrometer, an infrared spectrometer--these laser instruments provide the basis for in situ "remote" sensing of the sub-surface Martian environment within a powerful scientific package. The moles carry the instruments' sensor head array to the sub surface. The moles are thus required to undergo a one-way trip down the boreholes without the need for recovery of moles or samples, eliminating much of the robotic complexity invoked by such operations. PMID- 15754477 TI - [New research on the prevention and diagnosis of tuberculosis. Belgrade, 19 September 2003]. PMID- 15754478 TI - [The basis streamlines of activities of Department of Preventive Medicine of RAMS]. AB - The article gives a brief account of the main streamlines and scope of scientific activities of Department of Preventive Medicine of RAMS for the recent 10 years. PMID- 15754479 TI - [Hygienic estimation a state of nutrition of infant and preschool children age of city of Murmansk]. AB - The present research was directed on study of an actual meal and status of nutrition of children in the age of from birth till 5 years living in Murmansk (region of Far North). 998 children were surveyed. At an estimation of an actual meal of children the data about breast feeding are received, the basic nutrients misbalance of structure of diets of children are established, and their reasons are analyzed. On the basis of the received data the regional recommendations for organization of a healthy meal in children's preschool establishments and program of hygienic training of the parents to skills of a balanced diet of children of early and junior age were developed. PMID- 15754480 TI - [Combined spectral analysis method to trace the chemical elemental composition of food]. AB - We have applied the combined approach to the routine elemental analysis of food using the complex ICP AES + ICP-MS for both major and trace element groups. Matrix elements (up to 15-20), such as Na, Mg, Al, Si, P, K, Ca, Fe, Zn in biological samples can be determined by ICP-AES while the rest elements except of H, C, O, F, Cl measured by more powerful ICP-MS method, which is able to quantify up to 50-60 elements, depending on element/isotope abundances. The application of two methods provides more flexibility when working with unknown matrices and provides cross-checking to improve data reliability and also enhances sample throughput. PMID- 15754481 TI - [Contents of vitamin C, E, B6, beta-carotene and food fibers in of mushrooms]. AB - For specification and entering of the data in to Tables of a chemical compound of foodstuff definition of the content of vitamins C, E, B6 beta-carotene and dietary fibers in some kinds of the mushrooms collected in woods of Moscow suburbs was carried out. The received data testify to high enough content in them of vitamin C, especiale in white mushrooms and "fox" 24.0 and 28.8 mg%. The content of vitamin E in terms of a tocopherol-equivalent has made from 0.03 to 0.98 mg%. PMID- 15754482 TI - [Effect of food diet supplements with chromium on the clinical and metabolic parameters in type 2 diabetic patients]. AB - It was investigated the influence of food diet supplements with chromium on dynamic of glycaemia, lipid profile, blood pressure and weight in type 2 diabetic patients. Traditional hypocaloric diet was supplemented with chromium-spirulina (50 mcg chromium per day). The results investigations indicated that a chromium enriched diet has beneficial effects on basal and postprandial glycaemia, the content of cholesterol and triglycerides in serum in compared with a traditional hypocaloric diet. PMID- 15754483 TI - [The effect of diet with the sour-milk product enriched with amaranth on parameters of antioxigen protection at patients with a cardiovascular pathology]. AB - Investigation of influence of an antisclerotic diet with a probiotical sour-milk product enriched with an extract of leaves of an amaranth in patients with ishemic heart disease and hypertension was conducted. As a result of a diet were positive dynamic of clinic manifestation, lipid spectrum of blood, coagulogramms and antioxigen status. PMID- 15754484 TI - [The usage of food supplement to on the basis of calcium alginate for correction of liver infringements at virus hepatitis]. AB - The efficiency of food supplement (BAS) on the basis of calcium alginate was studied among the diseased people with virus hepatitis. BAS was taken 3.0 g one time in morning on an empty stomach 40-60 vin before eating or taking medicine. The results of clinical observations showed that BAS helps to improve the early normalization of clinical situation and biochemical indexes in blood with virus liver diseases. PMID- 15754485 TI - [Artificial feeding in clinical practice: modern status of the problem]. AB - We reviewed systematically and aggregated statistically the results of prospective randomized clinical trials to examine the relations among the nutrition interventions, complications, and mortality rates. We conducted a search for prospective randomized clinical trials comparing the effects of enteral and parenteral nutrition in adults. Two different people abstracted data for the method and outcomes separately. We used fixed-effects meta-analysis technique to combine the relative risks of the outcomes of infection, nutrition support complications, other complications, and mortality. PMID- 15754486 TI - [Food supplements and medicines of vegetative origin. Safety estimation and standardization]. AB - The article describes the modern legislative and normative base of both Russian made and foreign made food supplements turnover. The main requirements to them. The reasons for banning the use of some particular ingredients in food supplements formula are substantiated. The ways of improving the normative base for food supplements as well as for medicines of vegetative origin are proposed. The conclusion is made, that this base is more perfect than for the medicines, produced from herbs. PMID- 15754487 TI - [The main landmarks of formation and development of pathomorphological service of the Institute of Nutrition]. AB - This is a brief account of the main stages of formation and development of the pathomorphological service of the Institute of Nutrition the Russian Academy of Medical Science setting in 1930 year present time. PMID- 15754488 TI - [Methods of quantitative iodine analysis in food products and food sources]. AB - Methods of quantitative iodine analysis in food products and food sources. Advantages and disadvantages of the methods are evaluated methods of iodine analysis. PMID- 15754489 TI - Vitamin B12 status and its association with Helicobacter pylori infection in alcohol dependent patients. AB - Both infection with Helicobacter pylori and alcohol abuse have been associated with low vitamin B12 serum levels. The interaction between both risk factors is unknown. The aim of this study was to determine whether Helicobacter pylori infection is associated with low vitamin B12 levels in alcohol dependent patients. Blood samples were obtained from adult alcohol dependent patients undergoing detoxification and analyzed for serum vitamin B12 levels. Helicobacter pylori infection was serologically measured. Patient characteristics, medication use and alcohol consumption at admission were assessed by interview. A total of 6 out of 89 patients included presented low vitamin B12 levels, all were sub clinical deficient (<250 pmol/L) and none were clinical deficient (<150 pmol/L). Infection with Helicobacter pylori was present in 29% of the patients. The average vitamin B12 levels in Helicobacter pylori seropositive and seronegative patients were 1,033 pmol/L (SD 741) and 971 pmol/L (SD 717), respectively. The relation between Helicobacter pylori infection and vitamin B12 deficiency was not of significance (OR=0.48; 95% CI [0.05-4.32]). In conclusion, Helicobacter pylori infection is not a risk factor for low vitamin B12 levels in alcohol dependent patients. PMID- 15754490 TI - Dietary habits and selenium intake of residents in mountain and coastal communities in Japan. AB - We used a Simple Food Frequency Questionnaire (SFFQ) in combination with other dietary approaches to estimate the selenium intake from different food groups based on the average long-term diet, in two rural communities in Japan, one in a mountain area and the other in a coastal area. The intake frequencies of rice and wheat products were significantly different in the two districts. The intake frequencies of fish, meat, and eggs, which are rich in selenium, were not significantly different. The mean dietary selenium intake, estimated from the SFFQ and the 24-h recall method, was 82.7 microg/d (n=234) (range 19.2-180.1 microg/d) in the mountain community. The mean dietary selenium intake estimated from the SFFQ and average value of the normal portion size was 118.0 microg/d (n=123) (range 22.6-255.3 microg/d) in the coastal community. These estimated mean values exceeded the Japanese RDA, although the range of daily selenium intake was large. In the mountain community, fish made the largest contribution to dietary selenium intake (48.2% of daily total), followed by eggs (24.3%), and meat (17.0%). In the coastal community, fish accounted for 57.7% of daily total selenium intake, followed by meat (17.5%), and eggs (16.1%). In both districts, the total contribution of rice and wheat products was around 10%. It was found that the contribution of fish to dietary selenium intake was high and the contribution of cereals was low among Japanese. PMID- 15754491 TI - Effects of chicken extract on plasma antioxidative status and lipid oxidation in healthy rats. AB - Chicken extract has been consumed in oriental countries for centuries for improving body conditions such as recovery from fatigue. It is a rich source of antioxidant dipeptides. The in vivo antioxidative abilities were evaluated. Diets mixed with 4 different amounts of chicken extract were investigated for in vivo antioxidation ability using healthy male Sprague-Dawley (SD) rats. Total antioxidant status (TAS), thiobarbituric reactive substances (TBARS), iron content, superoxide dismutase (SOD) activity, glutathion peroxidase (GPx) activity and uric acid content were determined. In healthy rats, most of the indexes were not affected by intake of chicken extract significantly. However, plasma TBARS in the chicken extract-fed groups increased at the end of the experiment, which could be due to some pro-oxidative minerals in the extract. In conclusion, we found no significant or minor changes on the activities of antioxidative enzymes, antioxidant conditions, or lipid oxidation in healthy rats from consuming chicken extract, which may be the result of a balanced body condition. However, because of its high content of dipeptides, we suggest that it should have liver protecting effects if oxidative stresses are introduced. PMID- 15754492 TI - Effects of chicken extract on antioxidative status and liver protection under oxidative stress. AB - Chicken extract contains carnosine and anserine, both of which possess some antioxidant abilities. The objective of this study was to investigate the protective effects of chicken extract in male Sprague-Dawley (SD) rats under induced oxidative stress. Carbon tetrachloride was used as the oxidative stress inducer. Glutamic-oxalacetatic transaminase (GOT), glutamic-pyruvic transaminase (GPT), total antioxidant status (TAS), thiobarbituric-reactive substances (TBARS), iron content, and the activities of antioxidant enzymes were determined. We concluded that under oxidative stress, the intake of chicken extract was helpful in promoting the activities of antioxidant enzymes and in protecting the liver from oxidative damage. PMID- 15754493 TI - Effect of Lactobacillus rhamnosus KY-3 and cellobiose as synbiotics on lipid metabolism in rats. AB - Lactobacillus rhamnosus KY-3 is a fermentative bacterium that is used for the industrial production of L-lactic acid. We have examined the effect of L. rhamnosus KY-3 and cellobiose as synbiotics on lipid metabolism in rats. Rats were fed on a 20% casein diet (C) supplemented with either 1.7% L. rhamnosus KY-3 (KY-3), 10% cellobiose (CEB), or 1.7% L. rhamnosus KY-3 and 10% cellobiose (KY 3+CEB) for 13 d. The concentrations of serum total lipids, triacylglycerol, total cholesterol, and phospholipids were significantly reduced in rats fed a KY-3+CEB diet in comparison to those on the C, KY-3 and CEB diets. There was an increase in the weight of cecal contents and a significant increase in the amount of cecal short-chain fatty acids (SCFA). The dry weight of excretion increased additively upon the simultaneous administration of L. rhamnosus KY-3 and cellobiose (KY-3 + CEB). The amount of excreted fecal bile acids did not differ among the groups in this study. These findings support the hypothesis that the promotion of cecal fermentation can lower the level of serum lipids. These results suggest that simultaneous administration of L. rhamnosus KY-3 and cellobiose as synbiotics has a beneficial effect on lipid metabolism. PMID- 15754495 TI - The utility of anthropometric assessment at institutions and schools for individuals with intellectual disabilities and/or motor disabilities: a nation wide survey in Japan. AB - OBJECTIVES: To obtain information regarding the actual methods used for nutritional assessment at institutions and schools in order to establish a nutritional assessment method for individuals with disabilities. METHODS: Questionnaires were sent to 1,080 selected institutions and schools for individuals with intellectual disabilities (ID) and/or motor disabilities (MD). The response rate was 76.5%. RESULTS: The implementation rates for height and weight measurements were generally very high at both institutions and schools for individuals with ID and/or MD (85.5-100%), but those for other items were very low and varied among different disability types. The implementation rate for BMI was 17.9-71.9%, demonstrating that BMI was not widely used among institutions and schools for individuals with ID and/or MD. As for the methods for calculating percent body fat, a high percentage of institutions and schools for individuals with ID and/or MD indicated the use of bioelectrical impedance analysis for most disability types (60 - 77.9%). CONCLUSIONS: The percentages of institutions for individuals with ID and/or MD and of schools for individuals with ID and/or MD that implement nutritional assessment are very low, with variations among different disability types. PMID- 15754494 TI - Exercise training and supplementation with carnitine and antioxidants increases carnitine stores, triglyceride utilization, and endurance in exercising rats. AB - This study evaluated the effects of supplementation of carnitine and antioxidants on lipids, carnitine concentrations, and exercise endurance time in both trained and untrained rats as compared to non-supplemented rats. Thirty-two male SD rats, age 7 wk were divided into four groups according to exercise training and modified AIN-76 diets: NTNS (non-trained non-supplemented), NTS (non-trained supplemented), LTNS (long-trained non-supplemented) and LTS (long-trained supplemented). The trained rats were run on a treadmill for 60 min per day (10(0) incline, 20 m/min for 8 wk). Carnitine (0.5%/diet) and vitamin E (0.5 mg/g b.w.) were supplemented in rat diets and vitamin C (0.5 mg/g b.w.) and melatonin (1 microg/g b.w.) were administered into the stomachs of the rats. LTNS and LTS rats had significantly lower serum total lipid, triglyceride, total cholesterol and liver triglycerides, but had higher serum HDL-cholesterol. There were no changes in exercise endurance time by supplementation in untrained animals, however endurance times were longer in LTS animals than in LTNS. The supplementation and training tended to increase carnitine palmitoyltransferase (CPT-I) activities, although the differences were not statistically significant. Likewise, CPT-I mRNA levels were higher in both supplemented and exercise trained rats. These results suggest that supplementation of carnitine and antioxidants may improve lipid profiles and exercise ability in exercise-trained rats. PMID- 15754496 TI - Effects of capsinoid on serum and liver lipids in hyperlipidemic rats. AB - We compared the effects of capsinoid, a non-pungent component, on serum and liver lipids, with that of synthetic capsaicin in hyperlipidemic rats. Male Wistar rats of 9 wk old were divided into 4 groups: a control group receiving a high-fat diet containing 1% cholesterol, and capsaicin, capsinoid-I and capsinoid-II groups supplemented with 0.1 mmol of N-pelargonylvanillylamide (synthetic capsaicin), and 0.1 mmol and 1.0 mmol of capsinoid (capsiate: dihydrocapsiate=63 : 37) per kg of control diet, respectively. All groups were pair-fed for 4 wk. Compared with the control group, serum lipid levels in both capsinoid groups and liver lipid contents in the capsinoid-II group showed the same reduction as that of the capsaicin group. In the capsaicin and capsinoid-I groups, fatty acid synthase (FAS) activities were lower and hepatic triacylglycerol lipase (HTGL) [EC 3.1.1.3] activities tended to be higher than those of control group. Lipoprotein lipase (LPL) [EC 3.1.1.34] activity in adipose tissue was higher in the capsaicin and capsinoid-II groups than in the control group. These results showed that capsinoid can improve serum and liver lipid metabolism comparable to synthetic capsaicin. PMID- 15754497 TI - Changes in tissue protein synthesis are involved in regulating urea synthesis in rats given proteins of different quality. AB - The purpose of present study was to determine whether the regulation of urea synthesis is mediated through changes in supply of amino acids by protein synthesis and whether the concentration of ammonia, or activities of amino acid catabolizing enzymes, regulate urea synthesis when the dietary protein quality is manipulated. Experiments were done on three groups of rats given diets containing 10 g gluten, 10 g casein or 10 g whole egg protein/100 g for 10 d. The urinary excretion of urea, and the liver concentrations of glutamate, serine and alanine increased with a decrease in quality of dietary protein. The fractional and absolute rates of protein synthesis in tissues declined with the decrease in quality of dietary protein quality. The ammonia concentration in plasma and liver, and activities of hepatic amino acid catabolizing enzymes was not related to urea excretion under these conditions. These results suggest that the lower protein synthesis seen in tissues of rats given the lower quality of protein is likely to be one of the factors to increasing the supply of amino acids and stimulating urea synthesis. PMID- 15754498 TI - Low-salt O-miso produced from Koji fermentation of oncom improves redox state and cholesterolemia in rats more than low-salt soybean-miso. AB - The author prepared low-salt miso by koji fermentation using soy-oncom and okara oncom (9:1), at which time soybeans and okara were fermented with Neurospora intermedia (oncom miso, i.e. O-miso). Its usefulness as a seasoning, as well as high antioxidative activity in vitro and antimutagenicity have already been presented. In this study, the antioxidative activity of O-miso in vivo, as well as serum cholesterol-lowering action, were investigated concerning contribution to health. In rats fed O-miso, the serum alpha-tocopherol level and isoflavone aglycone-intake were higher than in rats fed low-salt soybean miso (S-miso). The activities of serum GSH-Px and hepatic catalase were also higher in rats fed O miso than in rats fed S-miso, while the TBARS values were lower in both the serum and livers of rats fed O-miso. Furthermore, O-miso intake suppressed a rise in serum cholesterol level and increased fecal bile acid excretion. Such a cholesterol-lowering action of O-miso may be attributable to the micelle-breaking or evacuant effects of indigestible proteinous residues and the antioxidative activity of isoflavone-aglycones. O-miso seems to be more beneficial to health than S-miso in view of these aspects. PMID- 15754499 TI - IgE-binding abilities of pentapeptides, QQPFP and PQQPF, in wheat gliadin. AB - Wheat gliadin is known mainly as the causative substance for enteropathy accompanied by diarrhea (celiac disease) and food-dependent exercise-induced anaphylaxis. However, little is known in regards to the allergenicity of gliadin in atopic dermatitis. In this study, the allergenicity of gliadin was demonstrated using sera of wheat-allergic patients with atopic dermatitis. Since there are many repeated sequences in gliadin, peptides containing each motif were synthesized to evaluate their IgE-binding abilities. As a result, QQPFP and PQQPF were found to be epitopes (minimum structures for the IgE-binding). Gliadin might crosslink patient's IgE-antibodies via these epitopes and trigger subsequent allergic reactions. PMID- 15754500 TI - A change in gastric mucosal ascorbic acid status with the formation, progression, and recovery of compound 48/80-induced acute gastric mucosal lesions in rats. AB - We examined whether gastric mucosal ascorbic acid status changes with the formation, progression, and recovery of acute gastric mucosal lesions in rats treated with compound 48/80, a mast cell degranulator. Fasted Wistar rats received a single intraperitoneal injection of compound 48/80 (0.75 mg/kg). Apparent gastric mucosal lesions occurred 0.5 h after compound 48/80 treatment, progressed gastric mucosal lesions were observed at 3 h, and a partial recovery of the progressed lesions was found at 6 h. The gastric mucosal concentrations of total and reduced ascorbic acids in compound 48/80-treated rats decreased to approximately 60% of the levels of untreated rats at 3 h after the treatment but the decreased concentrations of total and reduced ascorbic acids were almost completely returned to the levels of untreated rats at 6 h. The gastric mucosal concentration of oxidized ascorbic acid in compound 48/80-treated rats showed little change. The serum concentrations of total and reduced ascorbic acids in compound 48/80-treated rats increased at 0.5 h after the treatment and further increased at 3 h but the increased concentrations of total and reduced ascorbic acids were almost completely returned to the levels of untreated rats at 6 h. The serum concentration of oxidized ascorbic acid in compound 48/80-treated rats increased transiently at 0.5 h after the treatment. The hepatic concentrations of total. reduced, and oxidized ascorbic acids in compound 48/80-treated rats increased 3 h after the treatment, but these increases were not observed at 6 h. These results indicate that gastric mucosal ascorbic acid status is disrupted with the progression of acute gastric mucosal lesions in rats treated with compound 48/80. PMID- 15754501 TI - Effects of riboflavin on 6,7-dimethyl-8-ribityllumazine formation in growing cells of a riboflavin-adenine-deficient mutant, Bacillus subtilis. AB - The effects of riboflavin on the formation of 6,7-dimethyl-8-ribityllumazine (DMRL) were examined using the growing cells of a riboflavin, adenine deficient mutant, Bacillus subtilis. The administration of low concentrations (50-500 ng/mL) of riboflavin to the basal medium significantly inhibited the formation of DMRL without any change in growth. Especially, the supplementation of more than 250 ng/mL riboflavin completely restricted the formation of DMRL under the experimental conditions. The results clearly indicate that a rigid negative feedback inhibition is operative for the biosynthetic pathway of riboflavin in B. subtilis. PMID- 15754503 TI - [Expert standard pain management implemented: "For the first time I had no pain!"]. PMID- 15754502 TI - Hepatoprotective effects of the water extract from adzuki bean hulls on acetaminophen-induced damage in rat liver. AB - We examined the hepatoprotective effect of water-extract from adzuki bean (Vigna angularis) hulls on acetaminophen (AAP)-induced damage in rat liver. F344/DuCrj rats of 8 weeks of age were fed diets without and with 0.5% AAP or besides it 5% adzuki extract (lyophilized) on a daily basis over a period of 4 wk. At that time, serum aspartate aminotransferase activity in only AAP-treated group was higher than in both control and AAP plus adzuki extract (AAPA)-treated groups, while hepatic glutathione content and hepatic glutathione reductase and catalase activities in the AAP-treated group were lower than in the control group in contrast to the reverse in the AAPA-treated group. Hepatic phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide concentrations were higher in the AAP-treated group than in the control group, and were lower in the AAPA-treated group than in the AAP-treated group. Hepatic glutathione peroxidase activity was higher in the AAP-treated group than in the control group, although there was no significant difference between both AAP- and AAPA-treated groups in this respect. These findings suggest that the adzuki extract will serve as a prophylactic against oxidative damage to the liver. PMID- 15754504 TI - [Special nursing in urinary diversions: knowing the complications and preventing them]. PMID- 15754505 TI - [Venous catheter associated infection: possible prevention are often still not utilized]. PMID- 15754506 TI - [Nutrition in chemotherapy: preventing deficiencies]. PMID- 15754507 TI - [Cancer--what does it mean for the family? Communications should not be allowed to falter]. PMID- 15754508 TI - [Prevalence and course of fatigue in breast cancer: the kind of chemotherapy plays a role]. PMID- 15754509 TI - [Patient lack of knowledge about nausea in chemotherapy: becoming experts in their disease]. PMID- 15754510 TI - [Counseling concepts in nursing: closing the gap to the patient]. PMID- 15754511 TI - [Prevalence of decubitus ulcer in a longitudinal comparison: reliable data allow successful prevention]. PMID- 15754512 TI - [Emergency aid in Southeast Asia: rebuilding a standard of living (interview by Stefanie Lindl)]. PMID- 15754513 TI - [The self determination right--only a question of legal liability?]. PMID- 15754514 TI - [Euthanasia of patients by nurses: a fate with many causes]. PMID- 15754515 TI - [Psychology of severe illness: the end]. PMID- 15754516 TI - [From case to case: back to routine, despite it all]. PMID- 15754517 TI - ["Counselling" in nursing--an analysis of determining characteristics]. AB - Counselling plays an important role in different fields of activities within different professions. This study of the "Institut fur Medizin-/Pflegepadagogik und Pflegewissenchaft am Zentrum fur Human- und Gesundheitswissenschaften der Berliner Hochschulmedizin" has tried to identifiy determining characteristics of "counselling" within different disciplines and to analyse their transferability into the discipline of nursing. In the course of the investigation it appeared that some basic elements of counselling play a certain rolefor nursing. Nevertheless no profound concept for "counselling" exists yet. PMID- 15754518 TI - Beds in mental hospitals or beds in general hospitals, where should they be located? PMID- 15754519 TI - Another person's heart: magical and rational thinking in the psychological adaptation to heart transplantation. AB - BACKGROUND: The goal of this study was to examine heart transplant recipients' psychological adaptation to another person's heart, with particular emphasis on recipients' attitudes toward graft and donor. METHOD: Thirty-five male heart recipients were examined by: the Symptom Distress Checklist (revised) (SCL-90-R); the Depression Adjective Checklist (DACL); a Post-Traumatic Stress Disorder Questionnaire (PTSD-Q); a Heart Image Questionnaire (HIQ); and a Semi-Structured Interview (SSI), aimed at eliciting attitudes and fantasies regarding the transplanted heart. RESULTS: All instruments indicated high levels of stress even several years after the transplant, but, simultaneously, 73% of recipients felt that acquiring a new heart had had a dramatic influence on their lives with a new appreciation of the preciousness of life and a shift of priorities, toward altruism and spirituality. Sixty percent returned to work after the transplant but some had to adapt to a changed attitude from those around them who regarded them as anything from mystical creatures to vulnerable or still-sick individuals. While all recipients possessed a scientific knowledge of the anatomy and physiological significance of the heart (as revealed in the HIQ), many endorsed fantasies and displayed magical thinking: 46% of the recipients had fantasies about the donor's physical vigor and prowess, 40% expressed some guilt regarding the death of the donor, 34% entertained the possibility of acquiring qualities of the donor via the new heart. When asked to choose a most and least preferred imagined donor, 49% constructed their choices according to prejudices, desires, or fears related to ethnic, racial or sexual traits attributed to the donor. CONCLUSIONS: This study confirms the intuitive idea that heart transplant involves a stressful course of events that produces an amplified sense of the precariousness of existence. Simultaneously, it gives rise to rejoicing at having been granted a new lease on life and a clear sense of new priorities, especially with regard to relationships. Less expectedly, this study shows that, despite sophisticated knowledge of anatomy and physiology, almost half the heart recipients had an overt or covert notion of potentially acquiring some of the donor's personality characteristics along with the heart. The concomitance of the magical and the logical is not uncommon in many areas of human existence, and is probably enhanced by the symbolic nature of the heart, and maybe, also, by the persistent stress that requires an ongoing, emotionally intense, adaptation process. PMID- 15754520 TI - A community study on emotional distress among the elderly in Israel. AB - BACKGROUND: The elderly constitute a group at risk for elevated emotional distress. The Ministry of Health, the Central Bureau of Statistics and other national agencies jointly conducted a survey to investigate the health status of the elderly, including emotional distress and selective determinants. OBJECTIVE: This paper reports the distribution of the emotional distress mean scores by socio-demographic and health-related variables. METHOD: 5,055 ethnically diverse elderly residing in the community were interviewed with a questionnaire comprising socio-demographic and health promotion behavior items, in addition to the 12 item-GHQ scale. Univariate and multivariate methods of analyses were applied. RESULTS: Several variables that were associated with higher mean scores of emotional distress were identified, among them, female gender, living without a partner, and those with chronic medical conditions and impaired life-styles; 38% of the variance was explained by the study variables. CONCLUSION: Health planners and clinicians, including mental health consultants, may utilize these findings to develop individual and collective programs of intervention aimed at reducing emotional distress, particularly among the more vulnerable elderly. PMID- 15754521 TI - Controllability awareness and maladjusted personality traits: preliminary findings. AB - Previous evidence from studies with college students on three continents and with teachers in an Israeli border community under constant terrorist threat indicates that individuals who are higher in "controllability awareness" (CA, the extent to which an individual's responses to life situations reflect attention to distinctions between controllable and uncontrollable aspects of potential outcomes) perceive their lives as less stressful, report fewer physical symptoms, and think in ways that enable them to cope more effectively with environmental demands. The current study extends these findings to include clients seeking treatment in an outpatient institute for the treatment of psychological stress. Correlational findings from measures of CA, perceived stress and personality variables indicate that CA is associated with higher stress tolerance irrespective of clinical diagnosis and appears to mitigate a wide range of psychological difficulties. Clinical and research implications are discussed. PMID- 15754522 TI - Dialectical cotherapy. AB - In the dialectical cotherapy approach, cotherapists take opposing views and roles from the beginning phase of treatment until positive change has been realized. When coordinated and effectively carried out, their work mobilizes forces and neutralizes resistances and externalizes internal conflicts and ambivalencies, making them more readily available for therapeutic work and resolution. Theoretical formulations and anecdotal examples are offered to explain and demonstrate its effectiveness in psychotherapy. PMID- 15754523 TI - DSM-IV self-report and subjective evaluation by psychiatrists in Israel. AB - BACKGROUND: Psychiatric disorder, with the range of both subsyndromal and syndromal manifestation, is an important, yet often unrecognized and unacknowledged, problem among physicians. It is a subject that remains understudied, particularly among psychiatrists. The purpose of this study was to explore the subjective perception of mental illness among members of the psychiatric profession. METHOD: Psychiatrists attending an educational symposium completed, anonymously, a self-evaluation questionnaire in which they were asked to self-diagnose the presence of DSM-IV disorders. RESULTS: 110 responses were received (response rate: 52.1%); 90% of respondents indicated the presence of at least one syndrome or trait. The most common disorders on axis I and axis II were "mood disorder" and "narcissistic traits" respectively, with the least common being "psychotic disorder" and "schizotypal traits." Female psychiatrists reported more impairment, particularly among axis I disorders. The reported number of axis I and II conditions decreased with subjects' age. CONCLUSIONS: Manifestations of psychiatric conditions including the range of subthreshold phenomena, as self-diagnosed according to DSM-IV criteria, appear to be prominently reported, albeit with low severity, in a subjective manner by psychiatrists. Our findings may be of importance in encouraging the implementation of special programs in training and ongoing occupational support. PMID- 15754524 TI - Patients' satisfaction with psychiatric inpatient care. AB - BACKGROUND: Patient satisfaction with medical treatment is important, and although in mental health the literature on the topic is scant, awareness of the issue is rapidly growing. AIM: Evaluation of patient satisfaction among hospitalized psychiatric patients. METHODS: 100 consenting patients (47 men and 53 women) were surveyed and completed a questionnaire regarding satisfaction with psychiatric care. SETTING: Fifty-nine percent of the patients were in a new modern building, 41% were in an older hospital building; 21% were in a psycho geriatric ward. The various settings allowed comparison across different environments. RESULTS: Most participants expressed mild general satisfaction. The overall rating was 3.40 +/- 0.43 (range 1-5). The ratings were similar regarding the various components of treatment: satisfaction with the nursing staff, physical conditions on the ward and treatment per se. Positive correlation was found between satisfaction with nursing and overall ratings. CONCLUSION: Inpatients were mildly satisfied with treatment. However, no correlation was found between satisfaction and either the type of case manager or the physical conditions in the ward. Further research is needed to characterize specific variables that may influence patient satisfaction with psychiatric care. In addition, the question of whether self-report evaluation questionnaires are appropriate for the inpatient psychiatric population should be addressed. PMID- 15754525 TI - Psychiatric services in nursing homes in Israel: should the American legislation be adopted? AB - BACKGROUND: Nursing homes provide care for the elderly who require medical, nursing or rehabilitation services. Legislation for the public health model of mental health care for nursing home residents in the USA was enacted in 1987. OBJECTIVE: To determine whether the USA act regulating psychiatric care for nursing homes may be applied in Israel. METHODS: Publications analyzing the outcome of the USA regulations demonstrate improved care as reflected by decrease in restraints and better use of psychotropic compounds. The shortcomings as well as benefits of the USA legislation are tested as to their relevance to the specific economic, environmental and medical issues in Israel. CONCLUSIONS: The adoption of USA legal acts regulating nursing home residents' psychiatric care may not be feasible in Israel. However, quality of care in nursing homes can be significantly improved if such regulations were "tailored" to Israel's unique structure of nursing homes. PMID- 15754526 TI - Reversible nocturnal enuresis in children receiving SSRI with or without risperidone: presentation of five cases. AB - Selective serotonin re-uptake inhibitors are indicated in a group of psychiatric disor ders childhood. Data about their use in the treatment of enuresis also exist. Voiding disorders related to their use have been reported. We present five patients in whom bed-wetting was observed during a treatment with a serotonin re uptake inhibitor with or without risperidone, drawing attention to a possible side effect of this group of drugs. PMID- 15754527 TI - Successful multidisciplinary treatment in an adolescent case of rumination. AB - Rumination is a rare and complex disorder to treat. It involves repeated regurgitation, rechewing and reswallowing of food and occurs mainly among infants and mentally retarded children. Among adolescents and adults it is rather rare, but still might cause significant sequelae (depression, other eating disorders and social problems). We provide a detailed psychodynamic formulation in a non retarded adolescent with rumination (over a four year period). A multidisciplinary approach involving individual insight-oriented psychotherapy, group therapy, pharmacological therapy and a nutritional intervention has proved successful in abating the patient's symptoms. We propose that among adolescents and adults, childhood issues of neglect and feeding problems may be reactivated through the rumination symptom. The focus of this report is to address the importance of rumination, as well as the role of a multidisciplinary approach to such cases. PMID- 15754528 TI - The legacy of Silas Weir Mitchell. PMID- 15754529 TI - [Chronic obstructive lung disease/COPD. Main symptoms: cough, expectoration, (stress) dyspnea, impaired physical performance]. PMID- 15754530 TI - [Platinum compounds: metabolism, toxicity and supportive strategies]. AB - Although the leading platinum compounds, cisplatin, carboplatin, and oxaliplatin, share some structural similarities, there are marked differences between them in therapeutic uses, pharmacokinetics, and adverse effects profiles. Compared with cisplatin, carboplatin has inferior efficacy in germ-cell tumors, head and neck cancers, and bladder and esophageal carcinomas, whereas the two drugs appear to have comparable efficacy in ovarian cancer, extensive small-cell lung cancers (SCLC), and advanced non-small-cell lung cancers (NSCLC). Oxaliplatin belongs to the group of diaminocyclohexane (DACH) platinum compounds. It is the first platinum-based drug that has marked efficacy in colorectal cancer when given in combination with 5-fluorouracil and folinic acid. Nedaplatin has been registered in Japan, whereas other derivatives, like JM216 (which is the only orally available platinum derivative), ZD0473, BBR3464, and SPI-77 (a liposomal formulation of cisplatin), are still under investigation. The adverse effects of platinum compounds are reviewed together with possible prevention strategies. PMID- 15754531 TI - [Sonographic first trimester screening in Switzerland]. AB - The sonographic measurement of the nuchal translucency is already regarded as the most valuable screening parameter for chromosomal anomalies. Beside standardised examinations profound information and counselling of the pregnant women should be emphasised. With the improvement of the specific maternal risk calculation using the sonographic measurement of the nuchal translucency, the biochemical markers and the maternal age, unnecessary invasive examinations may be prevented and their overall number can be reduced significantly. PMID- 15754532 TI - [Endogenous neuroprotective mechanisms in multiple sclerosis]. PMID- 15754535 TI - [Horsetail plant (not only) for the kidney]. PMID- 15754534 TI - [Rapidly progressing vision loss. 40-year-old office worker. Retrobulbar neuritis]. PMID- 15754533 TI - [Anaphylactic reaction after Fe-injection]. AB - An anaphylactic reaction after intravenous Fe injection (Ferrum Hausmann) occurred in an 84-year-old female patient on treatment for iron deficiency anemia. After emergency admission to hospital, it emerged that she suffered an acute anteroseptal myocardial event, but could be mobilized without any problems on adequate cardiac medication. After ruling out any hyporegenerative pathology, the hemoglobin level of 101 g/L registered can probably be explained within the scope of mild renal insufficiency. Instead of the intravenous form of iron (III) sucrose (saccharate) complex (Ferrum Hausmann i.v. / Venofer), the attending general practitioner had administered parenteral iron replacement i.v. in the form of Ferrum Hausmann (iron (III) hydroxide dextran complex), which is intended for intramuscular administration. This is the reason why, firstly, the oral form of administration should be preferred over the parenteral and, secondly, parenteral replacement using the iron saccharate complex should be only initiated in the case of a pathological oral iron load test indicating an iron resorption disorder. PMID- 15754536 TI - [Selection of dialysis type--a crucial decision. Peritoneal dialysis is better for the patient and cheaper for the society]. PMID- 15754537 TI - [Pregnancy and lactation are not risk factors for osteoporosis or fractures]. AB - Observational and case control studies infer that a pregnancy and a period of lactation are followed by loss in bone mass of up to 5%. The reason for this loss is virtually impossible to conclude as so many factors known to influence the bone mass undergo changes during a pregnancy and lactation. The increased calcium demand, changed nutritional habits, reduced smoking and alcohol consumption seen in many women during these periods, the changes in body weight and fat content, the changed level of physical activity and the changed levels of hormones with potential to influence the bone metabolism could all influence the bone mass. Most studies also report that the deficit in "bone mass" normalises after weaning. Multiple pregnancies and long total duration of lactation can not be regarded as risk factors for osteoporosis and fragility fractures as most reports indicate that women with multiple pregnancies have similar or higher bone mass and similar or lower fracture incidence than their peers with no children. PMID- 15754538 TI - [New service for laboratory investigation of problematic cases. Four patients illustrate the value of EQUALIS]. AB - In 1996, the Endocrinology Group in EQUALIS, in collaboration with The Thyroid Unit at The Section of Endocrinology at Sahlgrenska University Hospital, introduced a "patient-related" quality assurance program. Serum samples from patients with established or suspected thyroid disorder and/or results from thyroid-related biochemical measurements which were at variance with the clinical presentation, were analyzed by the participants of an external quality assurance program in endocrinology. The results from this program were informative as regards capacity, in the individual laboratories, for the evaluation of analytical and/or biological factors causing unexpected results from the measurement of thyroid-related serum components. We now present four clinical cases, which had offered diagnostic problems. EQUALIS, in collaboration with the participants of the external quality assurance program in endocrinology and on the basis of experiences from this program, now offers a diagnostic service for clinical chemistry laboratories and physicians when they are confronted with unexpected results from the immunochemical measurement of analytes included in the program. PMID- 15754539 TI - [Laboratory thyroid disease diagnosis--interdisciplinary cooperation. International consensus in Swedish perspective]. AB - In 2003, The National Academy of Clinical Biochemistry published a consensus monograph on thyroid tests (available via Internet from 2002). The document, a result of international collaboration, describes the pathophysiological background and clinical application of the measurements as well as principles for their technical performance. Of value, and of definite clinical relevance, is the detailed description of factors affecting the results in health and disease. However, we lack a description of the sometimes large differences in values obtained with different generally available methods, as well as a presentation of the occurrence and evaluation of analytical interference and related technical factors affecting results. Readers with little experience from quantitative measurement in the clinical laboratory, therefore, are not stimulated to obtain a better understanding of causes and evaluation of unexpected results. PMID- 15754540 TI - [The somatizing patient. Frustrating for the physician not to be able to help because of insufficient resources according to a questionnaire]. PMID- 15754541 TI - [Indications that physical activity has a therapeutic effect in depression. Exercise a valuable complement to current antidepressive therapy]. PMID- 15754542 TI - [Swedish executive officers' frustration over working conditions]. PMID- 15754543 TI - [Refugee children and medical ethics: notification of physicians' errors should be initiated and precede loyalty]. PMID- 15754544 TI - [Physicians should not be investigators for the Swedish Migration Board]. PMID- 15754545 TI - [Fewer processes, more patients! Let us (re)gain the personal physician-patient contact]. PMID- 15754546 TI - [Issuing death certificates--a dangerous mission]. PMID- 15754547 TI - Munchausen Syndrome bx proxy and criminal prosecutions for child abuse. PMID- 15754549 TI - Human identification of victims of natural disasters. PMID- 15754548 TI - The disposal of human ashes. PMID- 15754550 TI - Kadian v Richards. PMID- 15754551 TI - Damages in medical litigation in New South Wales. AB - In the period 2001 to 2003, the New South Wales legislature enacted four Acts that impinge on the assessment of damages in litigation against health professionals. They are the Health Care Liability Act 2001 (NSW), the Civil Liability Act 2002 (NSW) (as originally enacted), the Civil Liability Amendment (Personal Responsibility) Act 2002 (NSW) and the Civil Liability Amendment Act 2003 (NSW). This article considers the principles on which damages are assessed in medical litigation and how those principles have been affected by these four enactments. It points out that each piece of legislation was retrospective in applying to events that occurred both before and after its commencement. However, proceedings already issued before a particular date were excluded in each case from the retrospective operation of the legislation. The article provides details of the relevant dates of operation of each of the statutes. PMID- 15754552 TI - Wrongful birth in New Zealand. AB - Wrongful birth cases have been a feature of the common law. In this article the author examines wrongful birth cases against the background of the New Zealand accident compensation scheme. Initially cases were accepted under the scheme, but after major changes to the legislation in 1992, wrongful birth cases were declined cover. The author argues that this should continue to be the case, and that, as a result, New Zealand courts will have to make the same policy decisions in this area as those made by other Commonwealth courts. PMID- 15754553 TI - Healthy law makes for healthy children: Cattanach v Melchior. AB - When courts are forced to consider issues surrounding birth and the sanctity of life, it is inevitable that divergence of judicial, academic and public opinion will result. However, the issue of whether parents can recover the expenses of rearing a healthy child has long vexed judges and commentators of law, ethics and medicine both in Australia and globally, with considerable disunity. A cogent example is the recent High Court of Australia decision in Cattanach v Melchior (2003) 215 CLR 1, where the court split four to three and handed down no less than six individual judgments. The case involved the birth of a healthy child following an unplanned pregnancy resulting from a failed surgical sterilisation. By allowing parents to recover the reasonable expenses of rearing an unintended child until the age of 18 years, the decision has provided some limited and temporary legal clarity to the issue of wrongful pregnancy in Australia. It is seen by some as a victory for the reproductive freedom of women and the rights of the child. However, with uncertainty remaining on the issue of wrongful life claims in Australia and with legislative changes in Queensland and New South Wales that partly reverse the High Court's decision, there remains doubt about the future of such claims in Australia. PMID- 15754554 TI - Fat and the law: who should take the blame? AB - The incidence of obesity in both adults and children is rising at a rapid rate in most developed countries, including in Australia. Some obese people are seeking to place the blame for their condition on the fast-food industry, as demonstrated by the recent litigation in the United States brought by two obese plaintiffs against McDonald's. This litigation was unsuccessful, and on existing Australian negligence principles any similar litigation commenced here is likely to suffer the same fate. Principles of personal responsibility, autonomy and free will should prevail to deny a negligence claim. The risk of obesity and concomitant health problems from eating fast food to excess is an obvious risk which the plaintiff should not have ignored and which he or she has voluntarily assumed. It is for the Australian Government, not the courts, to regulate the behaviour of the fast-food industry. The government should take action by requiring all major fast-food chains to label their products with nutritional information, and by imposing restrictions on the advertising of food to children. PMID- 15754555 TI - Risk reduction in general practice and the role of the receptionist. AB - Medical receptionists play a crucial role in any practice as they are usually the first points of contact for patients and the intermediaries through whom contacts with medical practitioners are made. This article reports the findings of a qualitative study of medical receptionists undertaken to explore their role in general practice, particularly in relation to activities involving direct patient assessment, monitoring, counselling and therapy. The findings highlight a number of significant issues in relation to the potential liability of receptionists, medical practitioners, medical centre owners and insurers. PMID- 15754556 TI - Pharmacist misconduct: the pitfalls of practice. AB - This article identifies the changing role of pharmacists in the provision of health care and analyses 78 complaints against pharmacists over a recent 12-year period in New South Wales, finding that the majority of complaints were in relation to the oversupply of particular medications, from which some pharmacists made significant financial gains. A number of other areas of complaint were identified, including issues in relation to the recording and labelling of medications and the roles and responsibilities of pharmacists, dispensing errors, fraud, and sexual misconduct. As the roles of pharmacists continue to expand, with the growth of "compounding chemists", and the suggestion that pharmacists are now performing basic medical tasks which were once the domain of a general practitioner, it is increasingly important that pharmacy take its place in any debates about the provision and regulation of health services. PMID- 15754557 TI - Justice in Medicare: recent changes to the public health care system. AB - This article sets out and examines a number of changes proposed by the Commonwealth Government to the Australian Medicare system as part of the 2003 2004 and 2004-2005 federal budgets, and the 2004 federal election campaign. In assessing the suitability of these reforms, the idea of justice is discussed. Health, as a basic good, is argued to be a matter of distributional and rectificatory justice. A number of popular material principles of justice are also examined and shown to be unsuited as sole determinants of health care resource allocation decisions. In light of this, various problems with the reforms are identified and improvements suggested. PMID- 15754558 TI - Investigations of complaints and quality of health care. AB - Malpractice law is frequently justified by the claim that it improves health care services but this belief remains untested. Using a multiple case study in 16 remote rural areas in New Zealand, this study examined the effects of formal quasi-judicial investigations on the quality of health care services. The study found that the fragile local health systems were damaged by the quasi-judicial investigations of the medical disciplinary body and became less efficient and less user-friendly. A few doctors left rural practice and were difficult to replace. The remaining health workers responded to the investigations in a negative manner, losing confidence, enthusiasm and motivation for work; they performed in a less efficient manner, working more slowly, setting up barriers to access, ordering more tests and referring more to secondary care. Complainants also appeared to have been disadvantaged as a consequence of having complained. PMID- 15754559 TI - Assisted reproductive technologies: professional and legal restrictions in Australian clinics. AB - The professional and legal regulation of assisted reproductive technologies (ART) in Australia is a vast maze of intersecting laws and guidelines which place restrictions on the provision of services such as infertility treatment, surrogacy, sex selection for social reasons, donor insemination, pre-implantation diagnosis and human embryo research. This study investigated the application of these restrictions on clinical practice in New South Wales, a relatively unregulated State, and Victoria, a relatively highly regulated State. The results of the survey indicate that the range of ART services in Victorian clinics was far more limited than in New South Wales clinics. The Victorian clinics uniformly restricted access of single and lesbian women and did not offer social sex selection procedures. The New South Wales clinics adopted different polices regarding these services. It was found that restrictive laws governing "social" issues have a significant impact on the availability of ART services and some respondents seemed unclear about the nature of restrictions and laws relevant to their work. It was also found that "reproductive tourism" is prevalent and restrictions were circumnavigated by patients with assistance from clinics. It was concluded that more evidence is required to evaluate regulation in this field of medicine. PMID- 15754560 TI - Possibility of being sued and the repercussions that such a suit can have on the professional and personal life of a doctor. PMID- 15754561 TI - Advocate for our patients. PMID- 15754562 TI - Revised quality indicator. PMID- 15754563 TI - Assessing and treating sleep problems in family caregivers of intensive care unit patients. PMID- 15754564 TI - Culturally-competent nursing care for American Indian clients in a critical care setting. PMID- 15754565 TI - End-of-life education in the pediatric intensive care unit. PMID- 15754566 TI - What important step must be considered before transferring a patient from one facility to another? AB - When a patient is in need of transport, it may be sudden and unexpected. It is important that a referring facility is aware of how to initiate the transport process so that there is no delay in transport and the patient receives the safest and most competent care. PMID- 15754567 TI - Weighing the evidence in your favor. PMID- 15754568 TI - Process helped gain acceptance for open visitation hours. PMID- 15754570 TI - [Comparison of fetal ECG recordings using STAN S21 analyser and external ECG of newborns, in relation to enzymatic ischaemic markers and acid-base parameters in umbilical blood]. AB - DESIGN: Neonatal hypoxia is often responsible for myocardial ischaemia. To evaluate cardiac involvement in neonates, fetal and neonatal ECG recordings have been performed and parameters indicating heart dysfunction have been analysed. OBJECTIVES: Our interest has been focused on analysis of fetal electrocardiogram recordings using STAN S21 analyser during labour, compared to newborns ECG recordings in the first 12 hours of life, in relation to enzymatic ischaemic markers and acid-base parameters in umbilical blood. MATERIALS AND METHODS: The data were collected from 8 term fetuses and neonates, continuously monitored in the first and second stage of labor. All ECG were recorded with a single spiral scalp electrode connected to the STAN S21 analyser. After birth, umbilical venous blood samples have been collected for determination of pH, base excess and concentrations of creatinine kinase (CK), its isoenzyme (CK-MB), myoglobin, lactate dehydrogenase (LDH), alanine transaminase, aspartate aminotransferase. In the first 12 hours of life, external neonatal ECG was performed. RESULTS: P,QRS and T waves have been present in all cases recorded. There was no suggestion of fetal hypoxia in any case. A little variety of QRS complex has been observed. In our study, we didn't noticed any pathological changes in ECG recordings. The QT interval, corrected by the Bazett formula was normal in all cases. Nevertheless correct ECG recordings, increase of enzymatic ischaemic markers (LDH, CK,CK-MB) has been observed in two cases, corresponding to the normal fetal outcome. PMID- 15754569 TI - [Impact of delivery type on EGF and IGF-1 concentration in umbilical blood of newborns and their mothers' milk]. AB - Peptide growth factors, including EGF and IGF-1, play a substantial role in child's growth and maturation processes. Growth factors found in mother's milk can modify development of the newborn and the infant, especially in the area of alimentary tract differentiation and maturation. OBJECTIVES: Study was undertaken to evaluate impact of the delivery type on EGF and IGF-1 concentration in the umbilical blood of newborns and their mothers' milk. MATERIAL AND METHODS: Sixty eight newborns and their mothers were examined. EGF and IGF-1 concentration was measured in serum of venous umbilical blood as well as in the mothers' milk collected on the first day of lactation. To measure EGF and IGF-1 concentration radioimmunoassay method (RIA) was applied. RESULTS: No correlation was found between the concentration of the two growth factors in umbilical blood and the delivery type. Neither significant difference was found between EGF concentration in the milk of the mothers who had spontaneous labour and the factor concentration in the milk of the mothers who had a caesarean section. Contrary to EGF, IGF-1 concentration in the milk of the mothers who had a caesarean section was lower than in the milk of the mothers who had a spontaneous delivery. However, significant differences were observed only in a group of mothers of newborns born at term. CONCLUSIONS: Caesarean section may disturb mechanisms of IGF-1 synthesis and release to mother's milk in the initial stage of lactation. PMID- 15754571 TI - [Anterior abdominal wall defects--retrospective analysis of fetuses diagnosed in the Department of Obstetrics & Gynecology of the Postgraduate Center of Medical Education between 1997 & 2002]. AB - OBJECTIVES: Our objectives were to determine the risk of aneuploidy and anatomic anomalies in fetuses with different kinds of abdominal wall defects. DESIGN: We retrospectively studied ultrasound reports of the fetuses with abdominal wall defects. MATERIALS AND METHODS: We analyzed 108 cases diagnosed during the last five years in our center. All fetuses underwent a detailed ultrasonographic survey and in majority of cases (78.8%) antenatal karyotyping was performed. We also analyzed gestational age at the first examination. RESULTS: 35 cases presented gastroschisis, 60 omphalocele, 7 ectopia cordis and 6 limb-body wall complex. Associated anomalies were detected in 48.1% of all cases. It was 22.9% in gastroschisis group, 58.3% in omphalocele group, 42.9% in ectopia cordis group and in all fetuses with limb-body wall complex. Chromosomal abnormalities were present in 13% of all cases; in 2.9% of gastroschisis group, 20% of omphalocele group, 16.7% of limb-body wall complex group and none of the fetuses with ectopia cordis. The median gestational age at the first examination was 24 weeks. CONCLUSIONS: In fetuses with abdominal wall defects detailed ultrasonographic survey and in majority of cases prenatal karyotyping should be done. Furthermore patients with fetus with such a defect should be diagnosed as early as possible in the referral center. PMID- 15754572 TI - [Course of twin pregnancies and labors in the 1st Department of Obstetrics and Gynecology, Medical University of Warsaw]. AB - DESIGN: The aim of the study was to analyze the course of twin pregnancies and labors in the I Department of Obstetrics and Gynaecology, Medical University of Warsaw. MATERIALS AND METHODS: 124 cases of multiple pregnancy and labor that occurred in the I Department of Obstetrics and Gynecology within the period from 1994 to 2001 were retrospectively analyzed. RESULTS: The study revealed high risk of premature labor, premature rupture of the membranes (PROM), intrauterine growth retardation (IUGR) and pregnancy induced hypertension (PIH) in twin pregnancy. High incidence of cesarean sections was seen both in premature and at term labors. Significant differences between the mean birth weight and Apgar score of I and II twin were observed. Differences in condition of the neonates were smaller in the group of twins born by cesarean section. CONCLUSIONS: The higher incidence of both obstetrical complications and cesarean sections is associated with twin pregnancies. An increased obstetrical risk and lower Apgar score are observed in case of vaginal delivery of the second twin. Cesarean section seems to improve obstetric results and diminish the differences in condition between the neonates. PMID- 15754573 TI - [Lipid index (WL) in the blood serum of women undergoing surgery in the perimenopausal age]. AB - OBJECTIVES: In the blood serum from women divided into 4 groups (healthy women aged 24 and 45 years and patients after the gynaecological surgery with sparing or radical resection of uterus) the lipids index (WL) was determined. MATERIAL AND METHODS: The WL index expresses the correlations among the HDL, LDL, triglycerides and A1 and B apolipoproteins. In the perimenopausal period in women the decrease in the lipids index WL was observed comparing to the women at age of 25. RESULTS: During the diseases indicated the necessity of gynecological surgery in women in the perimenopausal period further decrease in the lipids index WL was noticed. Contrary to the sparing operation, surgical intervention with radical resection of uterus causes the following WL decrease observed in 3rd month after it. CONCLUSION: It may suggest that these differences have hormonal background. PMID- 15754574 TI - [Angioarchitecture of the endometrial venous system of human myomatous uteri- scanning electron microscopy study]. AB - OBJECTIVES: The venous system of endometrium of myomatous uteri was examined. MATERIAL AND METHODS: The studies were carried out on 20 uteri obtained from autopsies. Corrosion casts were studied using scanning electron microscopy. RESULTS: The specimens revealed a chaotic network of tortuous and distended veins, venules and venous capillaries. The so-called venous lakes were observable in the whole length of the functional layer of the endometrium. CONCLUSIONS: Veins of the endometrium do not go in a company with arteries. No arterio-venous anastomoses were found. PMID- 15754575 TI - [The occurrence of leiomyoma cellulare in the surgical material in the Department of Obstetrics and Gynecology of the District Specialty Hospital in Radom]. AB - DESIGN: It has been decided to estimate frequency of occurrence of leiomyoma cellulare (LC) among women operated because of the myoma and to compare data with bibliography. MATERIALS AND METHODS: 14 cases of LC among 309 women operated in the District Specialty Hospital in Radom because of the uterine myoma, between January 2000 and June 2002 were analysed. Clinical symptoms, patients' age, parity, radiological images, macroscopic traits, type of operation and histopathological features were analysed. Histopathological diagnosis was established according to the valid criteria: mitotic index < 5 MF / 10 DPW, lack of necrosis, little or none atypia. Qualifications to the operation were set on the basis of the gynecological examination, ultrasound examination and histological estimation of the scrapings from the uterus. RESULTS: LC was recognised among 4.53% of the patients operated because of myoma. They were mostly single tumours, of 7.54 cm in diameter (on average) occurring among women aged 41-50. The youngest patient was 29 years old, the oldest--73. No significant differences in prevalence of LC according to parity were stated. Clinical symptoms, ultrasound examination and macroscopic image were typical of myoma and did not indicate suspicion of LC. The most frequent operation was myomectomy (50%) and hysterectomy including appendages (42.86%) CONCLUSIONS: LC is a relatively frequent subtype of myo-levicellular myoma, which can be diagnosed only by microscopic examination. Because of microscopic similarities to other stromic tumours of the body of the uterus and myo-levicellular sarcoma it is necessary to examine a larger number of specimens and to use valid criteria of recognition. PMID- 15754576 TI - [Monochorionic, diamniotic triplet pregnancy complicated by twin reversed arterial perfusion sequence. Postpartum autopsy of placenta and acardiac fetus]. AB - The twin reversed-arterial-perfusion sequence (TRAP) is a severe complication of monochorionic twin pregnancies characterized by the hemodynamic dependence of a "recipient" twin from a "pump" twin. The recipient twin exhibits lethal abnormalities including acardia and acephaly. The pump twin has a mortality rate of 50% as a result of high-output heart failure. We present a case of a 24-year old female, gravida 2, para 2, with monochorionic diamniotic triplet pregnancy. The sonographic examination at 18 weeks' gestation revealed acardiac-acephalus fetus. Reversed arterial perfusion sequence was confirmed with Doppler sonography. Postpartum autopsy examination of placenta and acardiac fetus (acardius anceps) was detailed described. PMID- 15754578 TI - [Leukemic reaction in the course of uterine myoma and urinary tract infection in pregnancy]. AB - A leukaemic reaction is an increase of leukocytes count in peripheral blood between 20-50 G/l with significant percentage of immature white blood cells: metamyeloblasts, myelocytes and even myeloblasts in blood smear. The leukaemic reactions during cancers and many infections diseases, especially in the pregnancy, by their clinical symptoms and laboratory changes can imitate acute and chronic leukaemias. In this article we introduce a pregnant woman with the leukaemic reaction with leucocyte count 51 G/l and a presence of immature white blood cells with myeloblasts in peripheral blood smear. This leukaemic reaction was caused by inflammatory process in the myoma uteri and the urinary tract. PMID- 15754577 TI - [Cervical pregnancy]. AB - Until quite recently the diagnosis of cervical pregnancy was an indication for hysterectomy, because of mortality among patients with cervical pregnancy, coming up to 60%. During last 15 years methods of conservative therapy has developed in order to avoid the necessity of hysterectomy and to keep patient's fertility. Among these the most promising method is using of methotrexate (MTX), which is now the most commonly used agent in case of conservative forms for the treatment of cervical pregnancies. It should be stressed successful in treatment of cervical pregnancy in women, who wish to remain fertile, depends on early and proper diagnosis. MTX can be administered up to 13 weeks of gestation, with the highest efficacy between 6-8 weeks of pregnancy. We reported a case of cervical pregnancy treated successfully with methotrexate therapy combined with curettage of the cervical canal and uterine cavity. Serial beta HCG serum determination and ultrasonography were used to diagnose the pregnancy and to monitor therapy. Treatment was effective and the patient reproductive capability was preserved. PMID- 15754579 TI - [Noninvasive methods used in prenatal diagnosis]. AB - Research in developing methods for noninvasive prenatal diagnosis has focused on the following: the isolation of fetal cells from the cervix, from the blood of pregnant women and the isolation of the fetal DNA. In the paper authors review the potential clinical diagnostic applications of fetal cells and fetal DNA analysis in maternal plasma or serum and fetal cells in cervical mucus for noninvasive prenatal diagnosis and screening of fetal aneuploidy, gene mutations, preeclampsia and preterm labor. PMID- 15754580 TI - [Triplets, quadruplets and more...]. AB - Theoretically, a woman could bear unlimited number of fetuses. Practically, the human being on the top of his phylogenetic spread, has chosen the singleton birth as its dominant. The anatomical constitution of the woman let her feed no more than two children at the same time. This were probably the reasons for considering the multiple birth as a nature phenomenon. With the time passing, there was a growing body of legends and misunderstandings, based on exagerated histories on their births. Based on world literature, the super multiple births are analyzed. PMID- 15754581 TI - [The use of interactive medical data transfer to increase effectiveness of a mass screening program]. AB - Cervical cancer is still called the most frequent neoplasm refer to women's genital tract (incidence rate 30-35/100,000/year). Scandinavian countries experiences have shown that effective screening program especially used multimedia (radio, TV, Internet) is one of the based elements, which should improve situation in prevention of cervical cancer. The aim of this study was an attempt to create medical Website including specialized and basic informations for doctors and patients about cervical and breast cancer, their prophylactic and treatment. An influence of these data to increase active participation in screening program was also analysed. Material covers a random sample of 1200 women (30-59 years of age) who were invited to participate in cytology screening in two edition of the program (1997-99, 2000-02). In the years of 2000-02 information about Website was added to the invitation. Medical Website was created as server information. A special questionnaire prepared by Sociology Department was distributed to 1200 women. In both group participation during two screening edition was compared and analysed. All data were statistically correlated. Website address was created: http://pkzr.ac.bialystok.pl. From the group of 1200 questioned (mean age--41.8) answered 1059 women (88.25%). 6.98% (n=74) have never used this Website but 20 of them have done cytology test after receiving an invitation which were posted to all 1200 women. The last of 985 women analysed Website. 80.4% (n=792) have received enough informations about cervical and breast cancer (4.67 point) and decided to participate in cervical cancer screening. From this group only 428 women actively took part in cytology exams in 1997-99. 189 women participated in the screening programme from the last 193 patients (19.6%) who analysed server informations (4.89 point). From 141 people who have not answered for the questionnaire 23 have done cytology test. Participation in the second edition of cervical cancer screening programme (1997 99) was 41.9%. It was really two times lower than in the same group who could used Internet and was estimated on 85.3%. Internet site- http://pkzr.ac.bialystok.pl seems to be a basic digital textbook about cervical and breast neoplastic disease and their prophylactic. It is also interactive transferring data source that could create healthy attitude to increase effectiveness of screening programmes. PMID- 15754582 TI - Unilateral stereotactic posteroventral globus pallidus internus pallidotomy for Parkinson's disease: surgical techniques and 2-year follow-up. AB - With the advent of levodopa (L-dopa) and the recognition of its striking effect on Parkinson's disease (PD), virtually all surgical procedures for PD ceased from the mid 1960s. However, there has been a resurgence of pallidotomy and other stereotactic procedures in the last two decades as physicians realized that most PD patients eventually face medical failure after long-term treatment with L dopa. Nine PD patients, three men and six women, with an average age of 62 years and disease duration of 13 years underwent unilateral globus pallidus internus (GPi) pallidotomy contralateral to the side with marked akinetic symptoms and drug-induced dyskinesia. All patients were evaluated using the Unified Parkinson's disease Rating Scale (UPDRS) after drug withdrawal and while taking their optimal medical regimen, preoperatively and 6, 12, and 24 months after surgery. There was significant improvement in activities of daily living and motor subscores as well as total UPDRS score in the "off" state at the 2-year follow-up, which mainly resulted from improvement in contralateral bradykinesia and rigidity. Significant improvements in contralateral akinetic symptoms and drug-induced dyskinesia were also observed in the "on" state and were sustained for at least 2 years. Ipsilateral and axial symptoms were not altered by unilateral GPi pallidotomy. The complications of surgery were generally well tolerated. One patient had a small postoperative asymptomatic hemorrhage identified by routine follow-up magnetic resonance imaging. Another two patients developed temporary sexual disinhibition and auditory hallucination, respectively, which resolved spontaneously 2 weeks after surgery. The effect of pallidotomy for alleviation of akinetic parkinsonism is modest but significant, and continues to be effective for at least 2 years. Further analytical studies, especially the correlation of clinical effects and lesion locations, are important not only to provide direct feedback for surgeons to examine the technical accuracy and but also to facilitate understanding of the pathophysiology of PD. PMID- 15754583 TI - Transthoracic or transhiatal resection for middle- and lower-third esophageal carcinoma? AB - Transthoracic esophagectomy (TTE) and transhiatal esophagectomy (THE) are two common methods of resection for esophageal cancer. Although many studies have been performed in Western countries, there are still controversies over which method is the better procedure. In this study, postoperative improvement in dysphagia and the degree of postoperative pain were compared. The cases of 50 patients undergoing TTE and 23 undergoing THE for esophageal cancer between March 1997 and October 2002 were retrospectively reviewed. The location of the lesion, clinical stage (TNM), operative time, operative blood loss, hospital stay, complications, number of lymph nodes dissected, and survival duration were recorded. Pre- and postoperative dysphagia scores and postoperative pain perception (using a visual analog scale) were analyzed. Preoperative clinical stage and lesion site were not significantly different in the TTE and THE groups. The operative time was longer and the number of lymph nodes removed was larger in the TTE group. However, there were no differences in operative blood loss, hospital stay, complications, survival duration, and improvement in dysphagia. Pain perception in the THE group was significantly better than that in the TTE group. THE is a safe and rapid procedure, with recovery and survival periods similar to those for TTE. Both patient groups enjoyed the same ability to eat. Therefore, THE is an acceptable alternative to TTE for patients with middle- and lower-third esophageal cancer. Moreover, THE caused much less postoperative pain than TTE, which made patients more comfortable. PMID- 15754584 TI - Effect of glycemic control on electrophysiologic changes of diabetic neuropathy in type 2 diabetic patients. AB - Diabetic neuropathy is a common complication of diabetes mellitus. Effective blood glucose control retards changes in nerve conduction velocity in type 1 diabetes. This study examined the relationship between glycemic control and electrophysiologic changes in diabetic neuropathy in 57 type 2 diabetic patients. Nerve conduction in the peroneal motor nerve, tibial motor nerve, and sural nerve were measured at study entry and at follow-up 24+/-3.12 months later. Changes in individual nerves are expressed as a percentage change (PC) and overall electrophysiologic changes are expressed as the sum of individual PCs. The PCs for peroneal motor nerve velocity, tibial motor nerve velocity, and sural nerve velocity were all lower in patients with a mean HbA1c of 8.5% or less compared with those in patients with a mean HbA1c of more than 8.5%, and SPCV (sum of PC in velocity) was significantly inversely correlated with mean HbA1c. However, there was no significant difference in SPCV in subjects with or without hypertension, hypertriglyceridemia, or low high-density lipoprotein cholesterol concentration. In conclusion, hyperglycemia is the most important etiology for electrophysiologic progression in type 2 diabetic patients. Furthermore, a mean HbA1c of more than 8.5% will result in significant deterioration in electrophysiology. PMID- 15754585 TI - Connecting rigidities of various precision attachments compared with the conical crown retained telescope. AB - The purpose of this investigation was to observe the connecting rigidity of various precision attachments and to compare their connecting rigidities with the conical crown retained telescope (CCT). The connecting rigidity of a retainer was assessed using the flexibility test to measure the mesial and distal end displacements. Four precision attachments were analyzed: the dovetail slide attachment beyeler, cylindrical slide attachment, Spang Stabilex and Mini SG. The CCT was used as the control. Although there were many statistically significant differences between the displacements with the various attachments, displacements when vertically loaded were very small: all mesial end displacements were within 3 microm and distal end displacements were 21.4 microm. The largest of the mesial end displacements when horizontally loaded was as large as 44.5 microm (dovetail slide beyeler), while the others were all below 16.5 microm. The same phenomena occurred with the distal end displacements when horizontally loaded: the largest was seen with the dovetail slide beyeler, followed sequentially by the Spang Stabilex, CCT, cylindrical slide, and the Mini SG. The distal displacement with the dovetail slide beyeler was as large as 75.2 microm; those with the others were all below 31.2 microm, with numerous statistically significant differences between the displacements with the various attachments. Thus, connecting rigidities of rigid precision attachments are very similar to CCT, and only the dovetail slide beyeler attachment is too weak to resist horizontal displacement force. PMID- 15754586 TI - Unstable angina with normal coronary angiography in hyperthyroidism: a case report. AB - Hyperthyroidism is associated with an increase in myocardial oxygen consumption that, due to an imbalance of oxygen demand and supply, can cause angina. However, subclinical hyperthyroidism rarely presents as chest pain in the resting state. Herein, we present a case of subclinical hyperthyroidism involving a 58-year-old male who complained of frequent chest tightness and typical electrocardiographic changes while in a resting state. Coronary angiography showed no significant lesion. Laboratory data showed that the patient suffered from hyperthyroidism, for which he was successfully treated with anti-thyroid agents. We are reminded that typical chest pain might be the first symptom of hyperthyroidism. PMID- 15754587 TI - Dengue virus-associated hemophagocytic syndrome and dyserythropoiesis: a case report. AB - A 33-year-old man had dengue hemorrhagic fever with initial presentation of fever, leukocytosis, and thrombocytopenia. The cause of the subsequent rapid decline in red cell counts without evidence of intravascular hemolysis or massive bleeding was confirmed as hemophagocytosis and dyserythropoiesis by bone marrow study. The patient recovered with supportive care and the bone marrow pattern was normal on repeated bone marrow study. To our knowledge, this is the first reported case of dengue virus-associated hemophagocytosis and dyserythropoiesis in Taiwan. Clinicians should consider that the occurrence of hemophagocytosis and dyserythropoiesis could be due to dengue virus infection. That this dengue virus infection was confirmed by a positive serology result at the convalescent stage but not at the acute symptomatic stage underlines the need for a second dengue serology study, as dengue infection can be missed due to an initial negative serology result. PMID- 15754588 TI - Anterior subtalar dislocation: a case report. AB - Anterior subtalar dislocations are extremely rare. To our knowledge, six cases have been reported in detail in the literature, but for only two of these was an anteroposterior view radiograph used to confirm the diagnosis. We report a case of anterior subtalar dislocation in which the posterior tibialis tendon was incarcerated in the talonavicular joint and that required an open reduction. We discuss the diagnosis, mechanism, and treatment. PMID- 15754589 TI - Unusual gas pattern in emphysematous cystitis: a case report. AB - We present a case of emphysematous cystitis in a 43-year-old female. The gas pattern was unusual because there was only gas accumulation in the urinary bladder without bladder wall thickening or intramural gas formation. The differential diagnosis included postoperative states, enterovesical fistula, and iatrogenic instrumentation. The prognosis of emphysematous cystitis is good after early diagnosis and prompt treatment with appropriate antibiotics, blood glucose control, and adequate urinary drainage. The patient was discharged on hospital day 5 with one additional week of oral antibiotic therapy. She was quite well at the time of writing and her diabetes was controlled with an oral hypoglycemic agent. PMID- 15754590 TI - Wagner and Darwin, Hanslick and D'Arcy: from the whirlpools of becoming to the mathematical beauty. PMID- 15754591 TI - New speciation theories. PMID- 15754592 TI - Functional dynamics of living systems and genetic engineering. AB - The discussion on Genetically Modified Organisms (GMO's) has been centred mainly on the nature and effects on economy, human health, environment, of the few transgenic plant lines present in the market in the last eight years. On the contrary, the present paper starts with a discussion of some of the relevant changes in our basic knowledge of the structure and dynamics of living systems in the last twenty years. Contemporary Biology is then compared with what may be called the "modern paradigm" of life sciences on which present day GMO's are conceptually based. Technical, environmental, social and economic problems deriving from the unexpected, persistent prevalence of the old fashioned modern vision of life in the "spirit of time" will be thoroughly discussed with a particular attention to the virtualisation process of GMO's and the effects of the prevalence over economic, social, environmental reality of their symbolic values. PMID- 15754593 TI - Distant non-chemical communication in various biological systems. AB - Communication is a natural ability of all living systems. It is very likely that various types of communication were evolved during evolution. While the communication by means of chemicals, direct contact or via organs of sense is under intensive study for a long time, alternative ways of interaction are still considered debatable, This review covers the topic of physically mediated communication in various biological systems. PMID- 15754594 TI - Reaction-diffusion equations for simulation of calcium signalling in cell systems. AB - Numerous experimental and theoretical studies have recently pointed to the importance of calcium signals and their propagation as waves of various kinds inside cells. This phenomenon has been particularly noted in fertilized egg cells. Ca2+ plays a fundamental role in these cells as it is capable of stimulating, by means of a first, large wave, the beginning of an organism's life at fertilization, immediately after sperm penetration. Furthermore, calcium is involved in numerous subsequent processes that are essential for the development of the future embryo, e.g. in contraction of cortical cytoplasm, protein synthesis and cell differentiation. Calcium waves, which are generated by self oscillating pacemakers and propagate in excitable media, have been observed in some types of egg cells after fertilization. These waves adopt different shapes according to their emission frequency, wavelength, velocity and curvature, and they can occur as solitary waves, target waves or spiral waves. The mathematical models that study the progress of these waves have been developed by means of partial differential equations of the "reaction-diffusion" type. This study will discuss some significant models of intracellular Ca2+ dynamics. Some preliminary considerations will then be made in order to develop a model that describes the propagation of Ca2+ waves in ascidian eggs. PMID- 15754595 TI - Ecological modelling: from the biosphere to the everyday data gathering; filling the gap. AB - Biosphere, according to many authors, can be considered as a system characterised by control mechanisms, which allow the maintenance of its stability. If the Biosphere is a complex object, the coming out of the equilibrium will have as a result a rapid change which cannot be controlled and whose results cannot be foresighted. Modelling is, for this reason, desperately needed. A general mathematical modelling suggested that the Biosphere evolution be characterised by periods of stability and instability, which are related to the connections between its components. These components, both living and not living, are connected by feedback relationships maintaining stability. From this model a general ecosystem modelling was developed which was first applied to a very peculiar situation i.e. that of the hydro-thermal marine vents. The hydrothermal ecosystem is, anyway, a very peculiar one and it can be considered also a micro biosphere with its own sources of energy not related to sun radiation. For this reason it was possible to make a correlation between these very peculiar ecosystems and the Biosphere. More difficult it is to relate the modelling of the light depending ecosystems, which are, on the contrary, of great interest because they are directly linked to the Biosphere and objects of quite everyday data gathering. Usually the description of ecosystems, which is proposed by the everyday operative task of collecting data, and which represents the widest source of data for modelling, is related to the quality of waters or air. In this way we have to find tools correlating the data routinely investigated and the general model proposed. Here we present the other side of the approach, a local investigation of the qualities of water, with the hope to make a proposal for the ecologists to carry on their job also taking into consideration the problem of ecosystems modelling. An evaluation study of the environmental condition of Fine River, based on the E.B.I. method (Extended Biotic Index) applied to macroinvertebrates was carried out in 2002. Seven sampling locations were selected along the main flow and two were selected on the two most important tributaries that flow into the Fine, the Savalano and the Pesciera-Marmolaio. On the upper part of the river two locations were selected, up-river the artificial reservoir of S. Luce, and five locations were selected in the intermediate and potamal part. 43 taxa of macroinvertebrates have been detected, among them 4 Plecoptera and many Ephemeroptera assembled by environmental variables. Chironomids have been detected in all the locations. The multivariate analysis with Multidimensional Scaling (MDS) applied to the samples of the potamal part location shows variability in the macroinvertebrates assemblage. The E.B.I. method resulted in a good quality of the water on the upper part of Fine River. The E.B.I. results, corresponding to the second Quality Class (Q. C.), were determined by the presence of the Plecoptera Chloroperla and several Ephemeroptera. These taxa are considered among the most sensitive to pollution. Downriver of Santa Luce reservoir, the water condition gets worse (3rd Q. C.), and this remains consistent along the intermediate and potamal part of the river, though it is never to be considered really bad. Water quality of the Savalano results between the second and third Q. C., while the situation of the Pesciera that flows waters of really bad quality into Fine River is to be considered serious. The F7 station (Polveroni), situated at the closing section of the basin, proves to be suitable to summarise the general conditions of the river. Finally a proposal to use the model developed for the hydrothermal vents for a provisional modelling to recover the water quality in the investigated ecosystem is presented and discussed. PMID- 15754597 TI - Fractal analysis of the uterine contractions. AB - The fractal dimension D may be calculated in many ways, since its strict definition, the Hausdorff definition is too complicated for practical estimation. In this paper we perform a comparative study often methods of fractal analysis of time series. In Benoit, a commercial program for fractal analysis, five methods of computing fractal dimension of time series (rescaled range analysis, power spectral analysis, roughness-length, variogram methods and wavelet method) are available. We have implemented some other algorithms for calculating D: Higuchi's fractal dimension, relative dispersion analysis, running fractal dimension, method based on mathematical morphology and method based on intensity differences. For biomedical signals results obtained by means of different algorithms are different, but consistent. PMID- 15754596 TI - Lysenko's contributions to biology and his tragedies. AB - Trofim Denisovich Lysenko's life and work have been much analyzed and discussed in the world's literature. It is well known that Lysenko is notorious and has been regarded as a charlatan. Less well known is that he once made greater contributions to Biology and has been misunderstood in some aspects. In this paper, Lysenko s contributions to plant physiology, genetics, agro-biology and evolutionary biology are briefly reviewed. His tragedies and mistakes, such as mixing science and politics, denying the existence of genes, failing to build up suitable scientific collectives for the metabolism-biochemical studies of heredity, as well as his theoretical one-sidedness, are also discussed, thus reconsidering the case of Lysenko from a comprehensive and objective viewpoint. PMID- 15754598 TI - A fast centrifuge method for nematocyst isolation from Pelagia noctiluca Forskal (Cnidaria: Scyphozoa). AB - Nematocyst isolation from surrounding tissue is an important step to characterize Cnidarian venom. Although several protocols have been used to extract venoms from cnidarian tissues, the complete isolation of nematocysts from tissue is still difficult. The goal of the present work was to evaluate the effectiveness of three different media, Percoll, Ficoll and Methylcellulose in isolating nematocysts from Pelagia noctiluca tentacles by centrifugation. The complete sedimentation of nematocysts and tissue fragments to the bottom of the test tubes was observed in Ficoll and Methylcellulose suspensions. The best result was obtained using a discontinuous density gradient of Percoll: three types of nematocysts were concentrated in three different fractions along the density gradient. Protein assay and preliminary chromatographic analyses confirmed these results. PMID- 15754599 TI - A specific exercise program and modification of postural alignment for treatment of cervicogenic headache: a case report. AB - STUDY DESIGN: Case report. OBJECTIVE: To describe an intervention approach consisting of a specific active-exercise program and modification of postural alignment for an individual with cervicogenic headache. BACKGROUND: The patient was a 46-year-old male with a 7-year history of cervicogenic headache. He reported constant symptoms with an average intensity of 5/10 on a visual analogue scale where 0 indicated no pain and 10 the worst pain imaginable. Average pain intensity in the week prior to the initial evaluation was 3/10 secondary to trigger point injections. The patient's headache symptoms worsened with activities that involved use of his arms and prolonged sitting. METHODS AND MEASURES: The patient was treated 7 times over a 3-month period. Impairments of alignment, muscle function, and movement of the cervical, scapulothoracic, and lumbar regions were identified. Outcome measurements included headache frequency, intensity, and the Neck Disability Index (NDI) questionnaire. Intervention included modification of alignment and movement during active cervical and upper extremity movements. The patient also received functional instructions focused on diminishing the effect of the weight of the upper extremities on the cervical spine. RESULTS: The patient reported a decrease in headache frequency and intensity (1 headache in 3 weeks, intensity 1/10) and a decrease in his NDI score from 31 (severe disability) to 11 (mild disability). The patient also demonstrated improvement in upper cervical joint mobility, cervical range of motion, scapular alignment, and scapulothoracic muscle strength. CONCLUSION: Interventions that included modification of alignment in the cervical, scapulothoracic, and lumbar region, along with instruction in a specific active exercise program to address movement impairments in these 3 regions, appeared to have been successful in relieving headaches and improving function in this patient. PMID- 15754600 TI - Effect of fluidotherapy on superficial radial nerve conduction and skin temperature. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVES: The purpose of this study was to examine the effects of the superficial heating modality, Fluidotherapy, on skin temperature and on sensory nerve action potential (SNAP) conduction latency and amplitude of the superficial radial nerve in healthy individuals. BACKGROUND: Fluidotherapy is a dry, superficial heating modality, which also provides tactile stimulation through the bombardment of air-fluidized cellulose particles. Previous literature has documented a direct relationship between skin temperature and neural conduction velocity; however, there is an absence of published research examining the effects of Fluidotherapy, and of tactile stimulation specifically, on neural conduction. METHODS AND MEASURES: Twenty-one subjects between the ages of 22 and 31 years (mean+/-SD, 25.5+/-0.7 years) and without prior history of diabetes, alcoholism, renal or metabolic dysfunction, current pregnancy, or heat sensitivity were invited to participate. Subjects completed an upper quarter screening exam and medical history form prior to participation. One group underwent heat (46.7 degrees C-48.9 degrees C) and tactile stimulation, a second group underwent tactile stimulation alone, while a third served as controls. Dependent variables were assessed at 3 intervals: before the intervention, immediately after the intervention, and 20 minutes after the intervention. All interventions were 20 minutes in length. RESULTS: A mixed 2-way analysis of variance indicated a significant interaction between time of neural conduction velocity assessment and treatment group for the dependent variables of sensory nerve action potential latency (P< .001) and skin temperature (P< .001). Appropriate post hoc tests were performed for simple effect comparisons. An inverse linear relationship existed between skin temperature and latency (r2 = .65; Pearson product coefficient, -.81). CONCLUSIONS: Fluidotherapy treatment, which combines the effects of heat and tactile stimulation, significantly elevated superficial skin temperature, while tactile stimulation alone and no treatment (control group) did not bring about a temperature change. As the superficial skin temperature increased, there was a concomitant decrease in the distal sensory latency of the superficial radial sensory nerve action potential. These results should be an important consideration for the clinician using superficial heating modalities. PMID- 15754602 TI - CMs should take the lead as JCAHO patient flow standards go into effect. PMID- 15754601 TI - The MedRisk Instrument for Measuring Patient Satisfaction With Physical Therapy Care: a psychometric analysis. AB - STUDY DESIGN: Psychometric evaluation of a cross-sectional survey. OBJECTIVES: To determine the validity of measures obtained from the MedRisk Instrument for Measuring Patient Satisfaction With Physical Therapy Care (MRPS) to differentiate between patient satisfaction with internal and external factors. BACKGROUND: Self report measures that sample a variety of items provide clinicians with an array of information that may assist in assessing patient satisfaction. An important measurement characteristic of these instruments is the ability to discriminate between different factors that may influence patient reports of satisfaction with care, ie, discriminant validity. In previous work, exploratory factor analysis suggested that the MRPS questionnaire has a 2-factor structure: "internal," relating to the patient-therapist interaction, and "external," describing nontherapist issues such as admissions and clinic environment. METHODS AND MEASURES: One thousand four hundred forty-nine adult patients completed the MRPS questionnaire upon finishing their course of outpatient physical therapy treatment. Discriminant validity of the 2-factor model was assessed using confirmatory factor analysis. The measures from the 2 factors were then evaluated for reliability by calculating the standard error of measurement (SEM), and for concurrent validity by correlating the mean score of the factors and individual items to global measures of satisfaction. RESULTS: Confirmatory factor analysis supported a good to excellent model fit for the internal factor (7 items) and external factor (3 items). The SEM for the 2 factors was 0.19 and 0.24, indicating a low degree of measurement error. Both factors had high significant correlation with global measures of satisfaction (internal, r = 0.83 and 0.80; external, r = 0.71 and 0.71). All individual items within the 2 factors had significant correlations with global measures ranging from r = 0.33 to 0.80. CONCLUSIONS: Our findings provide evidence of discriminant and concurrent validity of the 2-factor solution for the MRPS questionnaire for the sample that was tested. This 2-factor solution yields measures that are relatively free of error and may discriminate between internal and external factors influencing patient satisfaction. Patients who complete their course of physical therapy report that the professional interaction between the therapist and patient, especially the meaningful exchange of relevant information, is critical for patient satisfaction with care. The generalizability of our data to patients who do not complete their physical therapy care or who are receiving care in other health care environments is unknown. PMID- 15754603 TI - Throughput department reaps dramatic rewards. PMID- 15754604 TI - Team approach improves hospital's patient flow. PMID- 15754605 TI - Hospitalists, pharmacists partner to cut errors. PMID- 15754606 TI - VA gets high marks for preventive, chronic care. PMID- 15754607 TI - Hospital initiative sees higher vaccination rates. PMID- 15754608 TI - Root out causes of DP failures. PMID- 15754609 TI - The eye and I: a very happy couple. PMID- 15754610 TI - The eyes have it. PMID- 15754611 TI - Optometry on the march: why should the house of medicine care? PMID- 15754612 TI - Stem cells from somatic cell nuclear transfer: research to unlock the body's potential for self-repair. PMID- 15754613 TI - War paint, wagon trains and the next race. PMID- 15754614 TI - 2005 agenda for the House of Medicine. PMID- 15754615 TI - Highlights of the AMA interim 2004 meeting. PMID- 15754616 TI - The challenge of preventing vision loss from diabetic retinopathy. AB - Diabetes is present in 7% of Missourians. Another 52% are at risk. Periodic eye examinations are key to averting vision loss from diabetic retinopathy. Signs of diabetic retinopathy are evident long before vision loss and include microaneurysms, retinal hemorrhage, microvascular and venous caliber abnormalities and neovascularization. Loss of vascular integrity can lead to retinal edema and neovascular growth. Laser ablation of the peripheral retina can curb neovascular growth. It can also help stabilize macular edema. PMID- 15754617 TI - Repeated intervals of high-dose corticosteroid: an overlooked therapy in multiple sclerosis. AB - The rate of conversion to multiple sclerosis is about 6% per year for five years after the first episode of optic neuritis. While the new MS agents have garnered much attention as promising agents to prevent MS relapses, an overlooked therapy is pulse high-dose corticosteroid (10 mg per kg or greater) as an intervention to delay or prevent the development of MS. Data from Optic Neuritis Treatment Trial (ONTT) and other studies underscore the need to investigate the efficacy of high dose corticosteroid in MS. PMID- 15754618 TI - Preventing vision loss from glaucoma: putting data from clinical trials into practice. AB - Glaucoma is a group of ocular diseases that cause progressive changes in the optic nerve head and/or visual field loss. An elevated intraocular pressure (IOP) is often, but not always, present. Data from five large-scale clinical trials suggest that the greater the IOP reduction, the greater the protection against visual field loss. Corneal thickness is now recognized as an important predictor of glaucoma progression. The thinner the cornea is, the greater the risk of progression. PMID- 15754619 TI - Improving preschool vision screening programs. AB - Early detection of amblyopia is important, but many preschool-aged children do not receive vision screening. Factors that account for missed opportunities in detecting vision problems in early childhood include the difficulty of evaluating vision in an uncooperative child or in an infant and toddler. Methods need to be found to improve the low rate of preschool vision screening. In Missouri, a study has begun on preschool vision screening to find the most cost-effective method. PMID- 15754620 TI - Accommodating intraocular lenses--a solution for presbyopia. PMID- 15754621 TI - Ophthalmic topics for the non-ophthalmologist. AB - This article provides information for the medical practitioner who deals with patients receiving care for eye disease. It is written for the non ophthalmologist physician. The goal is to update the practitioner by discussing common ophthalmic issues that are encountered in everyday interactions with patients. Glaucoma, age-related macular degeneration, and treatment of refractive errors are emphasized in this summary article. PMID- 15754622 TI - Laparoscopic Nissen fundoplication in a community hospital: analysis of 202 patients. AB - We analyzed 202 patients undergoing Laparoscopic Nissen fundoplication. One hundred and three patients (90.6%) had typical reflux symptoms. Hiatal hernias were present in 84.6%. Most patients had erosive esophagitis; sixty-four patients (31.6%) had strictures. Complications occurred in 38 patients (18.8%), with major complications in 15 patients (7.4%). Major complications included bleeding in six, two requiring transfusion, slipped Nissen in three, pneumothorax in two, and esophageal leak in two patients. Eleven patients had postoperative dysphagia that responded to dilation therapy within three months. There was no mortality. PMID- 15754623 TI - Degenerative disc disease in childhood and adolescence. AB - Degenerative disk disease (DDD) is a common cause of low back pain and leg pain in children and infants. DDD is often unrecognized and under-diagnosed. This paper presents six cases of DDD in children and discusses the clinical presentation and pathophysiology of DDD. PMID- 15754624 TI - [Quality of life in patients with obstructive sleep apnea]. AB - Obstructive sleep apnea syndrome (OSA) is a condition affecting up to 5% of the population, in which episodes of upper airway obstruction lead to temporary cessation of airflow, disturbed sleep architecture and daily somnolence. The health consequences of OSA also include psychological and cognitive deficits, an increased risk of systemic and pulmonary hypertension, coronary disease, bradyarrhythmias and motor vehicle accidents. Symptoms and complications of OSA lead to a significant decrease of health-related quality of life (HRQOL) of affected patients. We review the current literature on HRQOL effects of OSA and its treatment. There is good evidence of beneficial effect of the continuous positive airway pressure (CPAP) therapy on the quality of life of patients with OSA. Improvements in HRQOL are most appreciable in patients with moderate to severe OSA, although they also seem to be present in selected patients with mild OSA. The effects of dental devices and surgical procedures on HRQOL of patients with OSA have not been studied in randomized, placebo-controlled trials. Health related quality of life has become one of the major outcome measures in patients with sleep apnea. Assessment of HRQOL has become a crucial part of any clinical study involving patients with OSA. PMID- 15754625 TI - [Long-term results after resection of IIIA-N2 non-small cell lung cancer and complementary mediastinal irradiation]. AB - The authors present a retrospective evaluation of long-term results of treatment of patients with IIIA-N2 non-small cell lung cancer (NSCLC), operated and subjected to complementary irradiation of mediastinum in the years 1987-1990. 60 patients were observed: 10 women and 50 men, aged 37-74 years (mean age 56). In all patients a radical resection of lung parenchyma with the primary tumour and mediastinal lymphadenectomy were performed. The one-year survival was 63% (38 patients), three-year survival--15% (9 patients) and five-year survival 13% (8 patients). It was found that the five year-survival in patients with IIIA-N2 NSCLC, in whom a combined therapy was applied (surgery and irradiation), was low, and the most common cause of death in the first three years of follow-up were distant metastases. Moreover it was observed that the number of mediastinal lymph node groups had a great influence on the time of survival. PMID- 15754627 TI - [Spectrophotometric evaluation of nitrous oxide pollution in the work place of the anesthesiologic personnel in operating rooms]. AB - Chronic exposure to nitrous oxide and volatile anaesthetics can threat the health of the operation theatre personnel. In Poland there are no regulations determining the maximum concentration of anaesthetic gases in the air. The results of measurement of N2O pollution of theatre atmosphere presented in the article exceed European and North American standards. Lack of appropriate technical facilities in operation rooms is aggravated by incompetence of the technical supervision staff and negligence of the anaesthesiological personnel. It is important to enforce European law and standards of work safety in operation rooms. It is of double impact, health protecting and ecological. The authors suggest introducing activities aiming toward the improvement of work conditions in operation rooms and the reduction of uncontrolled release of harmful anaesthetics. PMID- 15754626 TI - [Urinary trace elements excretion in patients with cystine calculosis]. AB - Not all cystinuric patients develop urinary calculi. There is the need to identify modulating factors. In recent years the role of trace elements in the mechanism of stone formation has received a steadily increasing attention. The aim of our study was to determine urinary excretion of trace elements in patients with recurrent cystine calculosis. MATERIAL AND METHODS: 11 patients with recurrent cystine calculosis and 15 healthy volunteers participated in the study. Urinary zinc, copper, chromium, boron and arsenic concentrations were determined by use of inductively coupled plasma-mass spectrometry (ICP-MS). RESULTS: There was a statistically significant difference between the patients with cystinuria and controls in mean arsenic urinary excretion (0.625+/-0.6 microg/kg body weight/24 h vs. 1.264+/-0.635 microg/kg b.w./24 h, p=0.02). There was also a noticeable difference in urinary chromium, the mean concentration of which was higher in case of the patients with cystinuria (3.32+/-3.59 microg/kg/24 h) than in the controls (1.38+/-0.69 microg/kg/24 h). The mean urinary excretion of zinc, copper, and boron (in microg/kg b.w./24 h) were respectively 7.40+/-7.98; 1.33+/ 0.48; 26.55+/-15.81 in the patients with cystinuria and 9.84+/-5.33; 1.16+/-0.84; 35.87+/-13.76 in the controls. CONCLUSION: It seems that a decreased excretion of arsenic may contribute to the development of cystine calculosis. PMID- 15754628 TI - [Incidence of haemolytic anaemia during combination therapy of chronic hepatitis C with interferon alpha-2b and ribavirin]. AB - OBJECTIVE: Analysis of incidence of haemolytic anaemia during INF-alpha + ribavirin (RBV) treatment of chronic hepatitis C (CHC). MATERIAL AND METHODS: 95 patients (M=64, F=31, mean age: 42.5 +/- 11.9) with CHC. All patients received IFN-alpha 3 MU three times a week and 1000-1200 mg of RBV daily. Basic haematological and biochemical parameters were checked. HCV-RNA (Cobas Amplicor, Roche) was checked before and after 6 months of therapy. RESULTS: After the first month of treatment a reduction of haemoglobin level occurred at 30% patients (not always characteristic for anaemia), an increase of the total bilirubin was observed in 16.8%, ferrum in 15.8%, while that of lactate dehydrogenase only in 22.1%. 37% of the patients required a reduction of ribavirin doses. No patient had treatment stopped because of side effects. CONCLUSIONS: Lower HGB, RBC, WBC, HCT and PLT levels and raised total and unconjugated bilirubin levels were the most frequent changes observed during the combined antiviral treatment. Improvement of haematological and biochemical parameters most often occurs spontaneously or after RBV dose reduction. Presence of haemolytic anaemia does not require stopping of RBV treatment, but only RBV dose reduction. At about 2/3 of patients who start therapy with interferon and RBV treatment has to be continued up to 48 weeks. PMID- 15754629 TI - [Role of interleukin-18, interleukin-1beta and its soluble receptor (sIL-1RII) in early and late Lyme borreliosis]. AB - Although borreliosis was first described as a separate entity more than 20 years ago its pathogenesis still remains unknown. In recent years the role of pro- and antiinflammatory cytokines in the pathogenesis of borreliosis has been discussed. The purpose of the present study was to evaluate the role of IL-1beta, IL-18 and sIL-1RII in the development of early and late stages of borreliosis. The study group consisted of 60 patients divided into 3 groups: patients with erythema migrans, Lyme arthritis and neuroborreliosis. In all groups serum levels of IL 1beta, IL-18 and sIL-1RII were determined and in the patients with neuroborreliosis additionally in cerebrospinal fluid (CSF). The levels of cytokines and sIL-1RII were measured before the start of treatment and after its termination. Before the treatment the levels of IL-1beta, IL-18 and sIL-1RII in serum and CSF were significantly higher in all studied groups compared with the control. After the treatment, despite the regression of the clinical symptoms and significant reduction of initially high levels of the cytokines and sIL-1RII, only the levels of IL-1beta in all patients and the serum level of IL-18 in the patients with neuroborreliosis were comparable with the values in the control group. It could suggest that the inflammatory process was not inhibited completely and confirms the role of IL-1beta, IL-18 and sIL-1RII in the pathogenesis of borreliosis. PMID- 15754630 TI - [Reference values of 24-hour ambulatory blood pressure in healthy children by height]. AB - The aim of the work was to prepare the percentile curves for blood pressure measured by 24-hour ambulatory blood pressure monitoring (ABPM) in children by height. MATERIAL AND METHODS: ABPM was performed in 852 healthy children aged 7 17 years (433 girls, 419 boys). Systolic (SBP) and diastolic (DBP) blood pressure was measured for 24 h, including the period of activity and rest, using BR-102 Schiller Poland LTD monitor and Quiet Tycos ABPM, USA. The mean number of readings over the 24-hour period was 58.4+/-4. RESULTS: The mean SBP in the group of girls with height 120-172 cm increased from 102.2+/-5.5 to 121.0+-7.8 mmHg, and in the group of boys with height 120-180 cm and over from 103.7+/-5.7 to 123.7+/-6.2 mmHg. A strong positive correlation between the height and SBP and weak with DBP was noted. The percentile charts for blood pressure in children aged 7-17 by height were prepared. CONCLUSIONS: The mean systolic blood pressure decreased at night more than diastolic blood pressure, independently of height. Systolic blood pressure was stronger correlated with the height than with the age. PMID- 15754631 TI - [Effect of palliative chemotherapy with topoisomerase inhibitor on the quality of life and survival of patients with advanced colorectal carcinoma]. AB - Among several topoisomerase I inhibitors topotecan is worth taking into account because of its little toxicity, good tolerance and quick excretion. The study compared functioning of bone marrow, liver and kidneys during and after the treatment, as well as survival period and quality of life of patients treated with topotecan and patients who underwent symptomatic treatment. 35 patients aged 43-71 of both sexes, with advanced stage of large intestine cancer (IV grade in TNM scale) were included in the study. 20 hospitalized patients were administered topotecan, topoisomerase I inhibitor, intravenously in 5-day courses (1.5 mg/m2 of body surface daily) repeated 5 times with 21-day intervals. Before the beginning of therapy and after it the following were conducted: medical check-ups as well as surveys and test research including groups of somatic and vegetative symptoms connected with the disease (based on the questionnaire QLQ-C30). 15 patients with the similar stage of the disease who had not agreed to palliative treatment with chemotherapy were treated only symptomatically. Applying antineoplastic treatment with topotecan contributes to the extension of the survival time by 4 months on average, and mortality during and after the treatment is lower than in the group of patients with the symptomatic treatment only. Antineoplastic treatment improves the quality of life and delays the disease progression. The quality of life is better until the preterminal period of the disease and enables patients maintaining motor activity and independence. PMID- 15754632 TI - [Is intensive functional insulin therapy the method of choice in newly diagnosed type-1 diabetes mellitus?]. AB - The aim of our study was a prospective evaluation of type 1 diabetic patients treated with intensive insulin therapy. We recruited 100 patients (62 males and 38 females) aged 24.3+/-6.2 years with newly diagnosed type 1 diabetes. The mean observation period was 5.2+/-1.5 years. Parameters of diabetes metabolic balance, occurrence of chronic complications and patients' knowledge about the disease and the methods of its treatment were evaluated. 68% of the patients controlled their glycaemia regularly before main meals and 53% of them had a diabetic diary. In the knowledge test 20% of the subjects reached < or = 11 points, 62% 11-17 points and 18% > 17 points (mean 14.4+/-3.2 points of maximal 20 to achieve). The mean result in the questionnaire of knowledge about the disease was 28.1+/-4.9 points. Fasting glycaemia was 7.2+/-3.4 mmol/l, 2h postprandial glycaemia 9.4+/-3.6 mmol/l, HbA1c 7.5+/-1.4%, the mean C-peptide level 0.9+/-0.4 ng/ml and the number of hypoglycaemic episodes was 6/individual/month. We observed a statistically significant correlation between the level of patients' knowledge and HbA1c (r= 0.31, p<0.05). Retinopathy and nephropathy were detected in 8 (9%) and 6 (6.8%) subjects respectively. The risk of microangiopathy was connected with low knowledge (RR: 5.67; 95% CI: 2.02-15.82, p<0.0002). The study confirms the crucial role of intensive insulin therapy and systematic patients' education concerning the disease in maintaining a good metabolic control and thus reducing the risk of diabetic vascular complications. PMID- 15754633 TI - [Enhancing the healing of venous crural ulceration after surgical treatment with high voltage stimulation]. AB - The objective of the study was to evaluate the influence of a high voltage stimulation on the process of healing of crural ulcerations after causal surgical treatment. Two comparative groups of patients, A and B, were made at random from patients with venous crural ulcers, who had been earlier operated by Babcock' method. The group A consisted of 12, and the group B--13 patients. The patients in two comparative groups were treated pharmacologically. Ulcerations at the patients in the group A were additionally treated with the high voltage stimulation. Changes of the area, length, width, and volume of the tissue defect after the high voltage stimulation were assessed. Weekly changes of the above parameters were determined as well. After the therapy there was a statistically significant decrease of the total and pus-covered areas, length, width and volume of ulcerations in both groups, while there was no statistically significant difference--except pus-covered areas--between the groups observed. Only the reduction of pus-covered area in the group A was statistically significantly faster than in the group B. PMID- 15754634 TI - [Non-Hodgkin lymphomas of the nasopharynx in children--diagnostic and therapeutic difficulties]. AB - About 7% of all childhood cancers comprise non-Hodgkin's lymphoma (NHL). NHL are heterogenous group of neoplasms deriving from lymphatic system (cell B and T). B cell NHL characterize by high malignancy, but coincidentally good reaction for treatment. In about 20% primary tumour is localised within head and neck, and nasopharynx lymphomas comprise 10%. This location maintains the biggest diagnostic and therapeutic difficulties because of tumours of this site proliferate in the region of frequent infections postponing a proper diagnosis and the local control after complete treatment is difficult. AIM OF THE STUDY: The authors analyse clinical symptoms before diagnosis, the incidence of nasopharynx lymphomas, histopathological type of neoplasm, clinical stage, results of treatment. MATERIAL AND METHODS: The study includes 97 patients who were treated because of lymphomas between 1993-2002. The character of clinical symptoms and their duration, histopathological type of lymphomas, results of treatment were analysed. RESULTS: The primary nasopharynx location was assessed in 9 patients (9.3%). Sex: 7 boys, 2 girls, age: 2-17 years. The duration which elapsed from initial clinical symptoms to diagnosis was 2-10 weeks. The histopathological assessment in 6 children was Burkitt lymphoma and in 3 children -Burkitt-like lymphoma. Metastases: CNS--1 patient, bone marrow--1 patient, abdomen--1 patient. Treatment was performed according to LMB-89 protocol. RESULTS: First complete remission--7 patients; second complete remission--2 patients. CONCLUSIONS: Lymphomas of nasopharynx cause diagnostic problems because of their early stage pseudo-inflammatory manifestation. Special attention should be paid to perform imaging studies (MRI/CT), which are useful in the reaching the proper diagnosis. The radiologic evaluation of primary lesion is still difficult. In the doubtful cases, the surgery and histopathological examination are necessary. PMID- 15754635 TI - [Early results of rehabilitation after endoprosthetic replacement of the knee joints]. AB - The purpose of this research was evaluation of magnetic field influence of low frequency on the process of rehabilitation of patients after endoprosthesis plasty of knee joints. The research evaluated usefulness of objective and subjective measurement methods to estimate progress of patients rehabilitation after endoprosthesis plasty of knee joints. The object of research were patients after endoprosthesis plasty of knee joints that were accepted for rehabilitation to the Unit of Motion Organs Rehabilitation of the Upper Silesian Rehabilitation Center "Repty" in Ustron. For the purpose of the rehabilitation process evaluation, the condition of patients was goniometry and the pendulum test were performed twice, i.e. at admission and then at termination of rehabilitation. A modification of the Lysholm Scale and a subjective knee questionnaire (VAS) were also used. On the ground of obtained results one can affirm that important role in therapeutic rehabilitation after endoprosthesis plasty of knee joint, plays adequately matched kinesitherapy, however one cannot confirm any influence of alternating magnetic field of selected parameters. Measurement methods both subjective and objective proved to be useful and effective in evaluation of rehabilitation progress in described cases. PMID- 15754636 TI - [Pleural mesothelioma--case with effusion in both pleural cavities]. AB - Etiology of the pleural exudate is not always easy to establish with the routine diagnostic procedures. We report the history of a 55-year-old man, driver- without evident occupational exposure to asbestos dust. Patient was treated in hospital because of recurrent bilateral sanguineous pleural fluid. Repeated basic laboratory examinations of pleural exudate did not contribute to establishing etiology of the disease. At the beginning of hospital stay antituberculosis therapy was applied but was unsuccessful. Rapid accumulation of the fluid, deterioration of general condition of the patient, presence of dysplasia in the cells of the sediment of the exudate indicated possibility of diagnosis of neoplasm of the pleura. Intravenous injections of cisplatin and intrapleural application of bleomycin did not influence, however, the course of the disease. Final diagnosis was possible only after pleural biopsy (with Abrams needle) was performed. Histopathologic examination of the specimen disclosed: malignant mesothelioma biphasic type. Patient died after 3 months of observations and attempt at treatment. PMID- 15754637 TI - [Pseudoasthmatic syndrome due to achalasia cardiae]. AB - The authors present a case of an 18-year-old woman, were referred to an allergologist because of a suspicion of bronchial asthma. She had complained of respiratory symptoms: chronic bronchitis for three years, cough and dyspnoea. Auscultation revealed wheezies and rales. Spirometry showed severe airway obstruction. Because of negative result of a reversibility test, ex iuvantibus treatment with prednisone was used but no results were obtained. The next step of diagnostic procedure was computed tomography which revealed the trachea compression by the dilated oesophagus. Finally she was found by us to have esophageal achalasia. Pharmacological treatment with nifedipine did not give an improvement. After operation the asthmatic symptoms disappeared and marked improvement was found in the flow-volume curve and in the chest radiogram. The demonstrated case proves that esophageal achalasia should be considered as one of the differential diagnoses of pathological respiratory symptoms suggesting asthma difficult to treat. PMID- 15754638 TI - [Unsuccessful treatment of pneumothorax which was associated with hilar tumor]. AB - The authors present a case of unsuccessful treatment of pneumothorax which was associated with hilar tumor. A typical underwater-seal system was installed in a 79-year-old patient, but the lung was still collapsed. Because of prolonged air leak, the patient was operated on. During the surgical procedure several bullae in the lung and the inoperable hilar carcinoma were diagnosed. Despite the absence of air leak, the failure of the lung to re-expand was noted postoperatively. However, recurrent air leak, subcutaneous emphysema and respiratory failure caused the patient's death. PMID- 15754639 TI - [Two cases of cystic fibrosis treated as bronchial asthma in children]. AB - Cystic fibrosis (CF) is the most frequent genetic disorder occurring in Caucasian, followed to preterm death. Clinical diagnosis of CF is set in infant or early childhood. Late diagnosis of CF is most often caused by non-typical course of the disease, rarely by misdiagnosis and not taken CF into account in differential diagnosis. In this paper we present two cases of misdiagnosis lung and digestive symptoms, treated as asthma bronchiale. PMID- 15754640 TI - [Atypical celiac disease in an adolescent girl--case report]. AB - Coeliac disease is characterized by life-long gluten intolerance. There are a wide variety of clinical presentations, which range from severe diarrhoea and weight loss to asymptomatic forms. The primary treatment for coeliac disease is the removal of gluten from the diet to prevent both immediate and long-term complications. The case of 16-year-old girl with coeliac disease was presented. At the age of 2 years the patient with impaired growth and abnormal stools was suspected to have coeliac disease. She experienced symptomatic improvement on gluten-free diet, but after 3 years the treatment was discontinued. The patient denied gastrointestinal or skin problems. At the age of 14 years Raynaud's phenomenon was observed for the first time. Two years later episodes of Raynaud's phenomenon involved all fingers and toes. Body mass index (BMI) was 23.8 kg/m2. Levels of free-carnitine, tocopherol, vitamin B12 were below normal limits and homocysteine level was increased. Antiendomysial IgA, antireticulin IgA, antigliadin IgA and IgG antibodies were positive. The duodenal mucosa showed total villous atrophy. Gluten free-diet and multivitamin supplementation provided some benefit in reducing Raynaud's phenomenon. The patient's well being has improved markedly. Atypical coeliac disease is usually seen in adolescents and adults in whom features of overt malabsorption are often absent. In cases of health problems occurring in persons with history of malabsorption syndrome in childhood suspicion of coeliac disease should be heightened and appropriate evaluation undertaken. PMID- 15754641 TI - [Pathologic splenic rupture in the course of non-Hodgkin's lymphoma]. AB - Pathologic rupture of the spleen is an uncommon but life-threatening complication of some malignant diseases. This paper presents a case of splenic rupture which occurred in a man with non-Hodgkin's lymphoma. We briefly discuss etiology, pathophysiology, symptoms and treatment of this complication. PMID- 15754642 TI - [Meningococcal meningitis--familial outbreak]. AB - Severe infections caused by Neisseria meningitidis, clinically appearing as meningitis or sepsis, are constantly noted in developed countries, including Poland. Because of rapid clinical course and mortality rate they continue to pose a serious clinical problem. Most cases are sporadic ones, caused by the contact with a healthy carrier of N. meningitidis. Familial and group outbreaks are rare, being responsible for about 1% of cases. However, in family members of patients with meningococcal infection the risk of developing the disease during the following week is about one hundred times higher than in the general population and they should receive an antibiotic prophylaxis. We describe a familial outbreak of meningococcal meningitis with typical clinical features, which appeared in two members of a family during the same day, with a case of acute pharyngitis most likely of the same etiology in the third person, a child. In that instance, practically simultaneous appearance of consecutive cases gave no time for prophylaxis. However, suspicion of meningococcal disease in the first patient contributed to the prompt hospitalization and diagnostics and proper treatment in the two following cases. PMID- 15754643 TI - [Leukapheresis in children with chronic myeloid leukemia and pulmonary leukostasis]. AB - We present two patients with chronic myeloid leukemia and hyperleukocytosis above 350 x 10(9)/l causing the symptoms of pulmonary leukostasis. Therapeutic leukapheresis procedures with concomitant chemotherapy were initiated within 12 hours from the diagnosis. After the therapy commencement the general condition of the patients systematically improved and leukocytosis gradually decreased from the second day of treatment with disappearance of pulmonary leukostasis symptoms. Thanks to application of such treatment procedures during the cytoreductive period, severe complications related to leukostasis could be prevented. PMID- 15754644 TI - [Therapeutic approach to patients with neurocardiogenic syncope]. AB - Vasovagal syncopes are the most common cause of loss of consciousness and if they occur frequently they contribute to the marked decrease of the quality of life and need treatment. One of important problems is the possibility of injury during the syncope. In older patients it can lead to the fractures of extremities, complications of which could be life-threatening. Another problem is the reaction of the surrounding people trying to help the unconscious by keeping him in the vertical position. It promotes hypotonia and could cause damage of organs such as brain or heart. The authors review the current literature on the management of patients with vasovagal syncope. It seems that the conservative approaches preferring patient reassurance and training methods are successful in syncope prevention and help avoid the side effects and complications of pharmacological therapy or pacemaker implantation. These last two methods should be reserved for patients with the most severe symptoms of the disease. PMID- 15754645 TI - [ICAM-1: structure, biological role and clinical significance]. AB - ICAM-1 is an immunoglobulin-like protein expressed on the surface of several cell types including endothelial cells and cells involved in the immune response. It plays an important role in the adhesion and migration of leukocytes to the sites of inflammation. ICAM-1 is expressed as membrane-bound and soluble form. The latter was detected in blood serum and cerebrospinal fluid in the course of infectious, autoimmune as well as proliferative diseases. This article describes the role of ICAM-1 in the pathogenesis of various disease entities and its clinical significance. PMID- 15754646 TI - [Effect of lead on the cardiovascular system]. AB - Lead is a metal widely spread in the natural environment. It is strongly toxic, particularly to the peripheral and central nervous systems. The toxic influence on the cardiovascular system is most pronounced in case of higher exposures, where myocardium and the renal circulation are affected, in consequence of which secondary arterial hypertension can develop. It seems that lead affects the cardiovascular system mainly by changing the peripheral autonomic nervous system and leading to chronic neuropathy. Chronic exposure, even to low doses of lead, can impair conduction in myocardium. In order to assess those changes thoroughly prospective studies involving newly employed workers with occupational exposure to toxic activity of lead will be necessary. PMID- 15754647 TI - [Techniques for creating hemodialysis fistulas]. AB - Hemodialysis is the primary method of renal replacement therapy. To conduct hemodialysis therapy, an effective vascular access is necessary. The authors describe basic types of surgically created accesses for hemodialysis and their complications. Peripheral blood vessels are listed by order in which they are used for access creation. PMID- 15754648 TI - [Effect of hyperthyreosis on the concentration of antioxidative vitamins and microelements]. AB - In the course of hyperthyreosis reactive oxygen production increases and oxidative stress develops. The consequences of oxidative stress comprise disturbances in balance of pro- and antioxidative agents, including changes in concentration of antioxidative vitamins and microelements. Although the results of published studies concerning changes of concentrations of vitamins and microelements in hyperthyreosis are contradictory, most frequently a decrease of zinc and magnesium concentrations was observed. PMID- 15754649 TI - [Melatonin and its biological significance]. AB - Melatonin is a hormone produced in human by the pineal body, the endocrine gland localized in the central part of cerebrum. It regulates many vital processes. Its main and best known effect is restoring the natural cycle of organism functions. It is safe and non-addictive sleep-inducing drug, which can eliminate disruptions in our circadian rhythm, in such situations as shift working, changing of time zones (during intercontinental air travelling) or insomnia. It improves mood and quality of sleep. Melatonin function consisting in stabilization of biological rhythms, free radical scavenging or immune system stimulating can delay aging processes. Its appropriate supplementation can prolong life even by decades, keeping our body in good both physical and psychological condition. Additionally, profitable for health properties of melatonin include ability to control some illnesses (prophylaxis of cardiovascular system diseases, neoplastic diseases and other functional disorders of organisms). It makes the immune system stronger, decreases susceptibility of the organism to stress, and improves mood and general feeling. PMID- 15754650 TI - [Melatonin and bio-elements]. AB - Melatonin occurs naturally in human organism, and its basic and the best investigated function is stabilization of biological rhythms, mainly day-night cycle. Properties very profitable for organisms, this hormone owes to its great ability of free radical scavenging and antioxidant activity. Connection between melatonin and elements occurring in human organism can be based on various relationships. Calcium ions play active role in melatonin biosynthesis, and sodium and potassium ions, thank to their properties of membranes polarization, influence melatonin penetrating possibility into cells, where this active molecule can exerts its physiologic functions. Another type of relationship exists between melatonin and zinc ions. Genomic action of melatonin appears in its ability to stimulate antioxidant genes expression, and its direct cellular action appears in increasing of these enzymes activity, among which very important place takes zinc-enzyme-superoxide dismutase (SOD). Melatonin also prevents poisoning by such elements like chromium (III) and (VI), iron and copper, through levelling toxic actions of these ions on organism. The role of melatonin in these processes relies mainly on scavenging of arisen free radicals, detoxification of hydrogen peroxide and combining excess of toxic ions into compounds harmless for organism. PMID- 15754651 TI - [Pneumocystis carinii--diagnosis based on DNA amplification technique]. AB - Pneumocystis carinii f. sp. hominis is an important cause morbidity and morality, causing P. carinii pneumonia (PCP) in AIDS and other immunocompromised patients. Long thought of as protozoan, molecular analysis of the ribosomal gene locus first demonstrated that P. carinii is a member of the fungal kingdom. Currently P. carinii is considered to be fungus likely to Ascomycetes, based on its ultrastructure, certain elements of its cellular biology, and nucleotide sequences of main genetic loci. The standard method for diagnosis P. carinii is microscopic examination of stained (immunofluorescent or conventional tinctorial) invasive specimens: BAL (bronchoalveolar lavage), lung biopsy or induced sputum with sensitivity varying range from <50% to >90%. Molecular detection system have the potential to provide a higher degree of sensitivity than microscopic method. PCR technique have been applied to lower respiratory tract clinical specimens and recently to non-invasive oral washes as well. Because P. carinii infection can be rapidly progressive and success of therapy dependent to the time of the initial therapy, early diagnosis and treatment is essential. PMID- 15754652 TI - [New aspects of epidemiology and treatment of Pneumocystis carinii infection]. AB - Pneumocystis carinii is an opportunistic eukaryotic pathogen most common causing pneumonia in immunocompromised host. DNA P. carinii f. sp. hominis has been frequently detected in air samples collected from environments of P. carinii pneumonia (PCP) patients and nasal swabs samples from contact health care workers and family. Epidemiological studies suggested not-only the possibility of person to-person infection transmission. First hypothesis of reactivation latent infection of the causing the new episode of PCP based on serological studies. Recently findings based molecular studies has suggested new acquired infection may to be cause of many cases of PCP. Early diagnosis, therapy and effective prophylaxis of PCP is a very important strategy to reduce morbidity and mortality among infected patients. The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is effective for prophylaxis and treatment P. carinii pneumonia. Empiric therapy in immunocompromised host also including AIDS patients with anti-Pneumocystis prophylaxis is also not recommended. Both dapsone and sulfamethoxazole act inhibiting the folate biosynthetic enzyme DHPS. Mutations identified in P. carinii in gene coding this enzyme are associated with resistance to sulfonamide. P. carinii may also developing resistance to atovaquone, second-line therapeutic and prophylactic agent. PMID- 15754653 TI - [Prevalence of Proteus mirabilis strains in clinical specimens and evaluation of their resistance to selected antibiotics]. AB - Proteus mirabilis are opportunistic microorganisms which cause serious hospital infections. The resistance of 1038 strains of P. mirabilis to selected antibiotics (lactams, aminoglycosides, fluoroquinolones, nitrofurantoin, cotrimoxazole, tetracycline) was tested by disk diffusion method. The least of P. mirabilis strains were resistant to imipenem and aztreonam. Most of P. mirabilis strains were isolated from urine, wounds and ulcers from patients of departments of urology, dermatology, general surgery and cardiosurgery. P. mirabilis strains were isolated from 4.3% of all positive samples. PMID- 15754654 TI - Smoking cessation--part I. PMID- 15754655 TI - An inaugural conference on occupational health in The Gambia: exploring the world through international occupational health programs. AB - This conference on workplace safety was a success for both the Gambians who attended and the Americans involved in planning, implementing, and funding the conference. The Americans were delighted to bring about some understanding of worker safety to a country that has struggled with labor issues for centuries. The dialogue reflected the concern that stakeholders in both countries have for labor issues, but there is a tremendous need for ongoing partnership with industrialized countries to bring these ideas and strategies to fruition. The Gambia has received some assistance in the area of economic development, but not in improving workplace safety. A wide range of economic support activities have been initiated by the ILO in The Gambia during the past 20 years. This has included providing guidance with international labor standards, assistance to the development of labor and related services, support to employer and worker organizations, vocational training, job creation and employment policy, and small enterprise development. Many challenges lie ahead for The Gambia in improving workplace safety. Serious concerns remain related to minimum safeguards in the workplace. For example, the Labor Act specifies safety equipment that an employer must provide for employees working in designated occupations, but these are rarely offered (Culp et al., 2003). Workers are plentiful and jobs are not, so workers tend to take any employment they can without regard to working conditions. The government has not formally revoked employers who have contributed to worker deaths. Industrial inspections are conducted, although there is little consequence for violations. The cycle of poverty, younger workers, and lack of education also contribute to many worker safety issues. Farm workers cannot read instructions on pesticide containers and industrial employees cannot understand workplace hazard communications. While the Gambian constitution mandates free compulsory primary education, the current state of educational opportunities is so dismal for poor families that many will not go beyond primary school and will enter the work force without technical skills. Teachers are overworked. While some young people could develop small business enterprises related to tourism and woodcarving, the vast majority continue a downward spiral into poverty (McGrath & King, 1999). PMID- 15754656 TI - Frequent flyer business travelers: major exposure hazards. AB - Bagshaw (2004) notes "the modern commercial aircraft cabin is maintained with adequate environmental control for the comfort of most healthy individuals" (p. 417). Occupational health nurses frequently deal with a population that may include unhealthy individuals or those with pre-existing conditions. It is critical for occupational health nurses to stay current with major hazards faced by frequent flyer business travelers to assist in identifying and preventing adverse health effects associated with these exposures. PMID- 15754657 TI - Benchmarking medical absence: measuring the impact of occupational health nursing. AB - Occupational health nurses are in a unique position to influence medical absence costs through their focus on health promotion and restoration. Corporate management looks to occupational health nurses to maximize employee productivity and reduce costs through lowered disability claims, fewer on-the-job injuries, and improved absentee rates. Benchmarking provides the occupational health nurse with the diagnostic tools to measure and manage productivity, demonstrate results, and justify the resources needed to accomplish desired outcomes. Benchmarking assists the occupational health nurse in all standards of practice outlined by the AAOHN: Assessment, diagnosis, outcome identification, planning, implementation, evaluation, resource management, professional development, collaboration, research, and ethics. PMID- 15754658 TI - Managing the common cold. PMID- 15754659 TI - The Canadian contribution to violence risk assessment: history and implications for current psychiatric practice. AB - Over the past quarter-century, Canadian researchers, clinical practitioners, and policy specialists have made several notable contributions to the broad field of violence risk assessment and management. In part, these contributions have been fostered by major changes in law over this period; in part, they have been spurred by findings from large-scale Canadian prediction--outcome studies. This paper offers references for a range of Canadian-inspired assessment schemes designed to evaluate psychopathy and potential for violence against others. PMID- 15754660 TI - The clinical use of risk assessment. AB - In this paper, we argue that risk assessment should be considered a part of daily clinical psychiatric practice. We discuss the advantages and disadvantages of various risk assessment procedures. In the event that a high risk for violence is present, we advise on strategies for discharging our duty to protect the public. Finally, by way of an illustrative case, we apply theory to practice and discuss the issues of risk management and risk reduction essential to a modern approach to psychiatry. PMID- 15754661 TI - The state of contemporary risk assessment research. AB - The focus on assessing dangerousness in routine psychiatric practice developed when relatively little was known about factors related to violence, and the accuracy of predicting violence was distinctly below chance. Since the 1990s, however, significant research attention has been directed toward factors related to violence and mental illness, as well as toward factors related to the accuracy of risk assessment techniques. Sociodemographic and environmental variables have been identified as significant predictors of violence, as has the presence of substance abuse. However, the data on specific mental health variables are somewhat mixed. Many studies point to a modest increased risk of violence associated with major mental illness and psychosis, whereas other noteworthy studies have failed to confirm such findings. Studies of the accuracy of risk assessments indicate that both actuarial and clinical methodologies perform better than chance, although the former achieve greater statistical accuracy. Despite ongoing controversies, risk management strategies that encompass the strengths and limitations of our present knowledge are available to clinicians. PMID- 15754662 TI - Community treatment orders: profile of a Canadian experience. AB - OBJECTIVE: This study reports the first published Canadian profile of a sample of psychiatric patients from the Royal Ottawa Hospital in Ottawa, Ontario, who were issued community treatment orders (CTOs). METHOD: We undertook a population study of sociodemographic and health care use patterns from January 2001 to September 2003, using a standardized information collection tool. RESULTS: The issuance of CTOs was associated with a statistically significant reduction in the number and length of hospital admissions and increased use of supportive community-based services and supportive housing. CONCLUSION: CTOs are effective tools for allowing patients to live in the least restrictive setting possible while they receive diverse services. They also effectively reduce rates and lengths of readmission to hospital. PMID- 15754663 TI - International dosage differences in fluoxetine clinical trials. AB - OBJECTIVE: International differences are thought to exist in dosages used by clinicians treating mood disorders. This study examined international dosage differences in antidepressant clinical trials, using a database formed and maintained as a component of a Cochrane review of comparative clinical trials of fluoxetine. METHODS: This systematic review included 132 studies. A detailed set of methodological features and results were abstracted from the original publications and entered into an electronic database. Mean and maximum fluoxetine dosages were compared across countries. To evaluate the dosages of comparison medications, a defined daily dosage (DDD) ratio was calculated as the trial mean dosage divided by the DDD for that drug. RESULTS: Both the maximum and mean dosages for fluoxetine and comparison medications were higher in trials conducted in the US (fluoxetine weighted mean dosage 49.18 mg; 95% CI, 41.30 to 57.05), compared with trials conducted in Europe (fluoxetine weighted mean dosage 29.98 mg; 95% CI, 25.28 to 34.68). Since most clinical trials were conducted in Europe or the US, we could not determine whether different dosages tended to be used in other regions. CONCLUSIONS: International differences in prescriber behaviour may influence, and in turn be influenced by, the conduct of clinical trials. It is difficult to reconcile such differences with the principles of evidence-based medicine. PMID- 15754664 TI - Panic-agoraphobic spectrum and light sensitivity in a general population sample in Italy. AB - OBJECTIVE: This study aimed to verify a possible correlation between panic symptoms and photosensitivity, not only in panic disorder (PD) but also in the panic-agoraphobic spectrum. METHOD: One hundred and sixty-nine healthy and drug free subjects completed the Structured Clinical Interview for Panic-Agoraphobic Spectrum-Lifetime version (SCI-PAS-Lifetime) and the Photosensitivity Assessment Questionnaire (PAQ). RESULTS: The SCI-PAS-Lifetime total score was positively correlated with the total score of the PAQ photophobia subdimension (r = 0.44; P < 0.001); the SCI-PAS-Lifetime total score was not significantly correlated with the photophilia subdimension. As photophobia increased, we observed significant score increases in all SCI-PAS-Lifetime domains. Bivariate correlation showed higher coefficient correlation between the panic-like symptoms domain and photophobia (r = 0.44; P < 0.001). CONCLUSIONS: A high total score in the SCI-PAS Lifetime, which denotes more typical features of the spectrum, is associated with a higher level of light sensitivity and intolerance toward bright stimuli. This finding reflects clinical evidence that widely documents photophobic behaviours in subjects with PD and the importance of light stimuli exposure during the onset and course of such a disorder. Bright stimulation seems to be relevant both in PD diagnosed according to current DSM criteria and in the entire panic-agoraphobic spectrum, from nuclear elements of the disorder through subclinical states to the normal condition. PMID- 15754665 TI - Psychotic disorders clinic and first-episode psychosis: a program evaluation. AB - BACKGROUND: There is growing recognition that people presenting with psychotic symptoms for the first time need specialized treatment. The Hamilton Health Sciences Corporation, McMaster Hospital, offers one such program, the Psychotic Disorders Clinic (PDC); it addresses some of the problems posed by long waiting lists, lack of family interventions, and long-term hospitalizations. The PDC is affiliated with the Department of Psychiatry and Behavioural Neurosciences at McMaster University. The program's goals are to provide comprehensive outpatient care and early interventions for persons in the early stages of illness and, consequently, to improve symptom control and functioning and reduce hospitalizations. Key service components include providing low dosages of antipsychotics, offering specialized family education, and supporting return to school and work settings. OBJECTIVES: This study compared outcomes before and after enrolment in the PDC to determine whether first-episode patients achieved improved symptom control and functioning and fewer hospitalizations. METHOD: For a 12-month period, we followed 40 patients, aged between 16 and 45 years, who experienced their first episode of psychotic illness between 1997 and 2000. Prospective longitudinal data were collected at baseline, 3, 6, and 12 months. Outcome measures included symptoms, global functioning, employment rates, duration of untreated psychosis, and number of bed-days. RESULTS: Of the patients, 37 completed the study at 6 months, and 31 at 12 months. Over the 12 months, significant improvements occurred in psychiatric symptoms (P < 0.001), global functioning (P < 0.001), and the mean number of hospital bed-days (P < 0.001). CONCLUSIONS: It is feasible for small outpatient services to provide early intervention strategies and obtain good outcomes among first-episode patients. PMID- 15754666 TI - Screening for mild cognitive impairment: comparing the SMMSE and the ABCS. AB - OBJECTIVE: To compare the sensitivity and specificity of the AB Cognitive Screen (ABCS) with the Standardized Mini-Mental State Examination (SMMSE) to differentiate normal cognition from mild cognitive impairment (MCI), especially when educational level and age are taken into account. METHOD: This cross sectional study took place at geriatric outpatient memory clinics. Participants were community-dwelling adults, aged 55 years or over, referred from primary care settings (a minority of participants were referred from specialists) for assessment of memory loss and age-matched control subjects with no complaint of memory loss. Each participant had the ABCS and the SMMSE administered in random order on the same day. RESULTS: Participants included 124 patients diagnosed with MCI and 111 with normal cognitive function. The ABCS showed a statistically significant difference between normal cognition and MCI (ABCS score 111.7 and 104.6 points, respectively, P < 0.001) for the whole group. This difference was significant with the ABCS, regardless of participants' age or education. There was a significant difference between normal cognition and MCI for SMMSE scores (SMMSE score 27.8 and 27.2 points, respectively, P = 0.040), but the differences were not significant when age and education were taken into account. Age and education were shown to affect the scores of both instruments except for the ABCS scores of MCI subjects, which were not significantly affected by education (P = 0.059). CONCLUSIONS: The ABCS is more sensitive than the SMMSE in differentiating normal cognition from MCI. The ABCS appears to be less influenced by education. It has improved clinical utility with a wider range of scoring gradations, reduced ceiling effects, and shorter scoring and administration times. PMID- 15754667 TI - Attention-deficit hyperactivity disorder with and without obsessive-compulsive behaviours: clinical characteristics, cognitive assessment, and risk factors. AB - OBJECTIVES: To determine the prevalence of obsessive--compulsive behaviours (OCB) in a clinical sample of children with attention-deficit hyperactivity disorder (ADHD) and to compare ADHD children (with and without OCB) with respect to clinical and cognitive characteristics and risk factors. METHOD: We compared ADHD children (n = 134) with and without OCB, and we compared all ADHD children with a group of normal control subjects (n = 26) on various clinical, cognitive, and inhibitory control measures; neurobiological and psychosocial risk factors; and family histories of psychiatric illness. RESULTS: Clinically significant OCB was identified in 11.2% (15/134) of children with ADHD. Comorbid OCB was associated with significantly increased perfectionism, improved teacher ratings of inattention, and a decreased likelihood of having a first-degree relative with ADHD. Comorbid ADHD and OCB may also be associated with increased oppositional behaviour at home and better inhibitory control, though these differences were not statistically significant. CONCLUSIONS: Comorbid OCB is common in clinically referred children with ADHD and is associated with increased impairment relative to ADHD alone. Further, children with ADHD and OCB may develop symptoms as the result of different genetic and environmental risk factors, compared with children with ADHD only. PMID- 15754668 TI - [Validation of the Peritraumatic Distress Inventory's French translation]. AB - OBJECTIVE: This article outlines the French translation and validation of the Peritraumatic Distress Inventory (PDI) in a psychotrauma outpatient sample. METHOD: A total of 127 French-speaking individuals were assessed from 2001 to 2002, at their first psychiatric visit for a traumatic event. An assessment at 3 months was offered by mail to control temporal stability. RESULTS: The PDI French validation has good internal consistency, with a 0.83 Cronbach's alpha coefficient. Test-retest shows a very satisfying temporal stability, with a 0.79 coefficient of intraclass correlation in a confidence interval between 0.61 and 0.89. The concurrent validity analyzed through the correlation with the Impact of Event Scale Revised (IES-R), the Peritraumatic Dissociation Questionnaire (PDEQ), and the General Health Questionnaire (GHQ) is very good. Further, the principal component analysis shows a bivariate solution that explains 45% of variance. CONCLUSION: The PDI French version has a satisfying psychometric validity. Moreover, the factor pattern in this translation is very close to that in the PDI's original version. PMID- 15754669 TI - A novel form of treatment resistance in anorexia nervosa. PMID- 15754670 TI - Effectiveness of risperidone in delirium. PMID- 15754672 TI - A renewed interest in day treatment. PMID- 15754671 TI - Hypokalemia from risperidone and quetiapine overdose. PMID- 15754673 TI - Oncology nurses look to the latest evidence to treat mucositis. PMID- 15754674 TI - Challenges of mucositis remain, but we are making progress. PMID- 15754675 TI - Iron-deficient patients craving ice fairly common behavior. PMID- 15754676 TI - 109th Congress commences, healthcare policy remains a top area for attention and action. PMID- 15754677 TI - [Surgery of colorectal cancer--how much training is required to acquire sufficient skills?]. PMID- 15754678 TI - [Skills by training. Education and case volume are strong prognostic factors in rectal cancer surgery]. AB - The TME project (TME = total mesorectal excision) demonstrated and probably enhanced a major shift in rectal cancer surgical practice in Stockholm with an increased centralisation and specialisation. As a result, local control and cancer-specific survival has been significantly improved. In addition, the frequency of APR (APR = abdominoperineal resection) declined. TME based surgery demands surgical skill, which can be achieved by participation in education programmes and increased by personal training and experience. Variability in patient outcome is seen also with TME based surgery and is mainly related to case volume, with better results obtained in patients treated by high-volume surgeons. A combined treatment modality approach, integrating the colorectalsurgeon, diagnostic radiologist, pathologist, medical and radiation oncologist is necessary to achieve optimal outcome. PMID- 15754679 TI - [Urinary catheters and antibiotic treatment. Guidelines adherence could be better]. AB - Indwelling urinary catheter is a well-known risk for urinary tract infection (UTI). In order to minimize the risk for UTI urinary catheters should be used with prudence. On April 24th 2002 we performed a point-prevalence study on the use of indwelling urinary catheters in 31 acute hospitals and 766 long-term-care facilities (LTCF) with 6,369 patients and 22,153 residents respectively. In the hospitals an average of 16.5% of the patients were treated with indwelling urinary catheters. The corresponding figure for LTCFs was 6.2%. Compared to 1994 there was an increase in the prevalence of urinary catheters in hospitals (from 11% to 16.5%). For LTCFs it was not possible to make such a comparison. Suprapubic catheters were used in 4% of the cases in hospitals and 10% in LTCFs. Closed drainage systems were used in about 75% both in hospitals and LTCFs. Approximately 10% of the patients/residents with indwelling urinary catheter were treated with antibiotics due to an UTI. Urinary culture was performed in only about half of those cases. To ensure patient safety hospital departments and LTCFs should, to our opinion, make enquiries on prevalence of indwelling urinary catheters, drainage systems and antibiotic treatment among their patients/residents on a regular basis. PMID- 15754680 TI - [Children with mild mental retardation in the special schools. "Top of the iceberg"?]. AB - Mild mental retardation (MMR), in UK learning disabilities, is defined as an IQ in the interval between 50-70 in combination with defined adaptive criteria (ICD 10 and DSM-IV). We have analysed the prevalence, the functional assessments, associated impairments and medical aspects of the children who attended grade 1-9 in the special schools for children with mild mental retardation in the northern part of Stockholm in 2001. The prevalence was 0.4%. We assume that this prevalence represents the "top of the ice-berg", i.e., comprising those with IQ's in the lower IQ interval (IQ 50-60), since 63% of the children in this MMR-group had been diagnosed already at the time of school-start or the 3 first years at school, 76% had been medically assessed and 45% had at least one additional neuroimpairment. The "true" prevalence of MMR in this age-group is not known. Our conclusion is that we have probably identified less than half of the total group with MMR and that the unidentified group--"those who are under the surface" run the risk of never having their cognitive dysfunctions identified. MMR has wide implications for the individual in today's society. That means that there is a need of clear guidelines for diagnosis and assessments. With such guidelines and with increasing knowledge and acceptance in the society, more children and families will be offered appropriate measures and support. PMID- 15754681 TI - [Adults with severe mental retardation--a 30-year follow-up. Physically integrated in the society, but mental problems are often neglected]. PMID- 15754682 TI - [Adapted exercise important after stroke. Acute and long-term effects of different training programs]. AB - Following stroke, persons experience deficits in motor control, reduced muscle strength, disuse atrophy, reduced cardiovascular fitness and elevated energy expenditure during locomotion. Recent exercise studies with few subjects, report beneficial outcomes after strength and low intensity aerobic exercise training. Progressive strength and aerobic exercise programmes from 3 to 6 months produced gains in functional recovery and health-related functional status such as motor function, peak isokinetic torque, balance, endurance, peak aerobic capacity and overall fitness without exacerbating spasticity. Increased access to community based physical activity programmes is recommended to prevent deconditioning and to improve health related quality of life in persons after stroke. Well functioning rehabilitation from acute care, through special rehabilitation units to community-based physical activity programmes is important. PMID- 15754683 TI - [Diabetes registry a key to better and better treatment outcome]. PMID- 15754684 TI - [Showing a little respect in a cruel world. What a Swedish general practitioner can accomplish in Darfur]. PMID- 15754685 TI - [Has the Medical Products Agency left the evidence-based medicine? The banning of SSRI to children and adolescents is unfounded]. PMID- 15754686 TI - [Criticism against the atenolol meta-analysis remains]. PMID- 15754687 TI - [Emphasizing shortages is not a factual basis for assessment]. PMID- 15754688 TI - [History helps us ahead]. PMID- 15754689 TI - [Trauma and emergency surgery. Organization and surgeons experience]. AB - BACKGROUND: To evaluate if combining operative treatment of patients with trauma and general surgery emergencies offers a good operative experience and can be a model for a Trauma Center organization, we compare our surgical experience with that of our general surgeons. METHODS: We reviewed records to determine number of operation, need of intensive care unit care for patients treated, the after hours practice by the trauma and emergency surgeons and general surgeons over a 1-year period at Ospedale Maggiore of Bologna. RESULTS: Emergency and trauma surgeons performed more operations per surgeons (133.7 vs 102.6) and managed more patients in intensive care unit than general surgeons. 51.8% of emergency and trauma operations were after hours. CONCLUSION: The care of trauma and emergency patients resulted in a breadth and scope of practice for trauma and emergency surgeons compared well with that of general surgeons but in a worse lifestyle. PMID- 15754690 TI - Bariatric Analysis and Reporting Outcome System (BAROS) following biliopancreatic diversion. AB - BACKGROUND: The Bariatric Analysis and Reporting Outcome System (BAROS) has been recently introduced to assess the modifications of weight, comorbidities and quality of life (QOL) after bariatric surgery, in order to achieve a standard for comparison in the treatment of obesity. This study reports the Authors' experience, analyzing with BAROS a consecutive series of morbidly obese patients. METHODS: From November 1998 to February 2001, 30 patients with morbid obesity underwent biliopancreatic diversion (BPD) in our Department. Patients were followed-up after 1, 3, 6, 9, 12 months and than yearly. Modifications of the Body Mass Index (BMI), percentage of the initial excess weight loss (IEW%L), comorbidities and type and incidence of complications were measured. The course and QOL were assessed using BAROS after a minimum follow-up of 18 months, or in absence of IEW%L modification at two consecutive scheduled visits. RESULTS: According to the BAROS, the outcome was classified as Excellent in 10% of cases, Very Good in 63.3%, Good in 20%, Fair in 6.7%; no patients had Failure course. Ninety-three percent of patients had resolution of at least one of their major comorbidities, and an improvement of the medical conditions was registered in all the cases. The QOL was greatly improved in 55%, improved in 35% and did not change in 10% of the patients. CONCLUSION: BPD provides effective weight loss, improvement or resolution of major co-morbidities and increases the QOL of morbidly obese patients. BAROS is an useful tool to assess the outcome after bariatric surgery. PMID- 15754691 TI - [Emergency surgery in elderly: experience gained in 198 cases over 80]. AB - PURPOSE: To evaluate our 2-year experience in the emergency surgical treatment of elderly people (aged > or = 80). METHOD: A retrospective review was conducted of 198 elderly patients admitted to Emergency Surgery Unit of the Ospedale Maggiore in Bologna from 01.07.2001 to 30.06.2003. RESULTS: All the Patients were submitted to emergency operations. Mean age was 84.8 (range 80-96); Female were 152, male 73. Preexisting condition, ASA scores and surgical procedures were recorded. The postoperative mortality was 17.1%. The mean length of stay in our Unit was 9.7 days; 93 patients needed rehabilitation facility at discharge. CONCLUSION: The Authors concluded that emergency surgery entails a high risk to the patients, high cost in hospital resources and rehabilitation facility. PMID- 15754692 TI - [Benign-thyroid nodule: what's therapy? Personal experience]. AB - INTRODUCTION: Solitary thyroid nodule is an important pathology with an incidence of 2-5% in Italian population. The diagnosis was based on clinical exam, laboratory tests and ultrasonographic evaluation. MATERIALS AND METHODS: The study undertook has the purpose to assess the usefulness between lobe isthmusectomy (with corresponding risk of possible relapse and complications for reintervention) and total thyroidectomy, (with corresponding post-surgical treatment complications), for benign thyroid disease with solitary nodule, on selected cases for disease and corresponding risk factors, in the retrospective clinical study, using laboratory tests and ultrasonographic evaluation. The study was conducted on 80 patients admitted from 1994 to 2000 with diagnosis of benign thyroid nodule and operated with lobeisthmusectomy. In seven patients the operation had to be converted in total thyroidectomy. RESULTS: Operative mortality was nil and long-term results at a mean follow-up of three years are encouraging. DISCUSSION: The main advantages of lobe-isthmusectomy for benign solitary thyroid nodule consist in less postoperative complications and less hospital stay. CONCLUSION: Therefore our experience has carried us to consider the lobectomy which treatment choice for all those benign thyroid diseases, with, solitary nodule. PMID- 15754693 TI - [Treatment of serious hepatic lacerations with Ton That Tung's technique]. AB - BACKGROUND: Serious hepatic lacerations are associated with an elevated mortality which depends on the consequent haemorrhage. The hepatic resection in emergency, according to Ton That Tung, seems to be the more appropriate operation for the rapidity of execution and the low mortality associated (11% out of 1000 operations). METHOD: According to Ton That Tung' technique, 12 operations of hepatic lobe and segment resections were executed after trauma caused by road accidents, firearm lesions, sidearm lesions and bleeding neoplasms. RESULTS: Among 12 patients operated on, 9 healed (75%) and 3 died (25%). CONCLUSIONS: The high mortality in the hepatic resections depends on the technique used. The anatomic resection, according to Bismuth, has an operative time longer than the time described by Ton That Tung, time that in emergency is important because the mortality depends not only from the blood lost, but, also the time in which the blood is lost. Hepatic resection according to Ton That Tung is the gold standard for the treatment of serious lacerations and hepatic bleeding neoplasms. PMID- 15754694 TI - [Non derivated surgery indications and limits in bile duct iatrogenic injuries treatment]. AB - Recently the incidence of iatrogenic bile duct injuries has increased proportionally to development of the laparoscopic surgery. The objective of this study is to determine the possible application of non derivated surgical repairs in iatrogenic bile duct injuries treatment of the above procedures. These surgical approaches range from simply placing a surgical drainage to liver transplantation. With reference to Strasberg's classification non derivated treatments are listed, furthermore are discussed and analysed their indications and limits. Owing to their experience and specific literature, the authors came to the following conclusions: 1) These surgical options represent an ideal reconstruction technique because they restore bile ducts continuity without altering physiological mechanisms to which these anatomical structures are appointed. 2) The indications of applying these methods are rare and usually limited to those cases in which the lesion has been recognized during operation and in the immediate post operative period. PMID- 15754695 TI - [The multidisciplinary management of acute pancreatitis: a review of 244 cases]. AB - BACKGROUND: Acute pancreatitis (AP) has still a high mortality rate; therefore the accuracy of the predictors of severity actually employed and the therapeutic choices are under debate. The aim of this work is to valuate the accuracy of the prognostic factors actually employed and the results of a multidisciplinary treatment of the AP. METHODS: A consecutive series of 244 patients affected by AP (168 mild, 76 severe) is analysed. Mean age 64.4 years (range 17-94 years old). As regard as aetiology is concerned, 166 are biliary pancreatitis, 42 alcoholic, 27 idiopathic, 9 iatrogenic. Ranson's score, Balthazar criteria, and ASA are employed to assess the high-risk patients. The treatment is medical and in severe AP the patient is admitted in ICU. 80 patients affected biliary AP undergo an ERCP. An endoscopic papillotomy is performed in 55 cases and in 49 biliary stones are removed. 90 patients underwent a delayed colcistectomy. 11 patients underwent a surgical treatment: 9 for infected necrotic pancreatitis, 1 for a hemorrhagic shock, 1 for peritonitis. The overall mortality, the mortality related to the severity of the AP, the prognostic evaluation of the etiology, the ASA index, the Ranson and Balthazar scores are evaluated. RESULTS: The overall mortality rate is 2.8% (0.6% in the mild AP and 7.8% in the severe AP). In the surgical group the mortality rate is 18.1%. The Ranson's score is the only independent factor in the multivariate analysis (p<0.0374) while also the Balthazar classification is significant in the univariate analysis. CONCLUSION: As regard as the prognostic factors in AP are concerned, the Ranson's score and Balthazar criteria are reliable even if many other predictors of severity are tested to make progress in the early detection of the complications. As regard as the therapy is concerned, the endoscopic papillotomy and the endoscopic removal of the biliary stones is the gold standard in the treatment of the biliary AP. In the other cases of AP medical treatment shall be started as early as possible with intensive care management in the severe AP. In patients with infected pancreatic necrosis or haemorrhage or peritonitis, surgery is the only therapeutic choice while the surgical treatment in case of sterile pancreatic necrosis with MODS unresponsive to medical therapy is still under debate. PMID- 15754696 TI - [Obstructing colon cancer--what's the surgical strategy?]. AB - INTRODUCTION: The aim of the study was to define the therapeutic approach used for obstructing colon cancer. PATIENTS AND METHODS: In the period 1990-2002 in the Emergency Surgery Department of the University of Naples Federico II, 81 patients underwent colon surgery for obstructing colon cancer (25 M, 57 F, age range: 55-88 years, mean: 72). Were performed subtotal colectomies, left or right hemicolectomies, sigmoid colectomies, anterior resections, temporary or permanent enterostomies. The authors put morbidity and mortality in touch with the type of surgical operation and with results of the literature. RESULTS: Mortality was 32%: 2.4% as a result of an anastomotic dehiscence; 4.9% for cardiovascular pathologies; 1.2% for sepsis; 23.4% for metastasis. Morbidity for anastomotic leakage was 4.9%: 2.4% after left emicolectomy and colo-colonic anastomosis; 1.2% after right emicolectomy and ileo-colonic anastomosis; 1.2% after defunctioning colostomy and left emicolectomy after 20 days. CONCLUSIONS: This experience suggests that a subtotal colectomy with primary anastomosis (one stage) can be performed more safely, by an experienced, skilled surgeon, in patients in good clinical conditions with acute obstruction of the colon. Healing remains a process depending more on the patient than on any aspect of the surgical technique. A defunctioning colostomy may be idealfor surgeons with little experience in colorectal surgery and in high risk patients with a very poor prognosis (unresectable lesions). PMID- 15754697 TI - Treatment of gunshot wounds to the colon: experience in a rural hospital during the civil war in Somalia. AB - BACKGROUND: In the last few decades there has been a clear tendency in civilian practice towards primary repair of gunshot wounds to the colon, resulting in a substantial decrease in the number of colostomies performed for this type of injury. METHODS: The series described here comprises 24 patients with gunshot wounds to the colon treated at the hospital of Jowar in the Middle Shebelle region of Somalia between 1999 and 2001. All injuries were caused by war arms firing high-velocity projectiles. RESULTS: In 18 patients surgery consisted of resection and immediate anastomosis, while in the remaining six patients colostomies were performed including five loop colostomies and one terminal colostomy. The sepsis-related mortality was 25% (6/24). CONCLUSION: Our experience had a peculiar setting, namely that of a civil war in a developing country. In this kind of "difficult" situation there probably exists no ideal procedure and the decisions taken at the operating table are influenced by various factors that are often related to subjective assessment. PMID- 15754698 TI - [Emergency surgery for neoplastic left colon obstruction: resection and primary anastomosis (RPA) versus Hartmann resection (HR)]. AB - This retrospective study compares: hospitalization time, morbidity and mortality (patients operated for neoplastic occlusion of the left colon in emergency, without perforation), considering hanestesiologic risk and surgical technique (RPA versus RH). MATERIALS AND METHODS: From April 1999 to February 2003 the Emergency Surgery of AORN of Caserta has operated, in urgency, 60 neoplastic patients with left colon occlusion realizing: 46 (76.5%) RPA [36 ASA < or = III, low hanestesiologic risk (LAR), 10 ASA = or > IV high hanestesiologic risk (HAR)], 12 (20%) RH (2 LAR and 10 HAR) 2 (3.5%) palliative colostomics. RESULTS: 36 LAR patients with RPA had 5% of specific complications and 5% of medical complications. 2 LAR patients with R-H: no complication. 10 HAR patient with RPA brought 10% of specific complications and 10% of medical complications. 10 HAR patients with RH had 20% of specific complications and 10% of general complications. The middle hospitalization of LAR patients with RPA was 11 days versus 9 days of RH pz. (equal hanestesiologic risk ). The HAR pz., treated with RPA, had as middle hospitalization 15 ggs in comparison to the 9 ggs of the HRA pz. treated with RH. MORTALITY: 1/36 for LAR pz. with RPA (3%), and 0/0 for LAR pz. with RH; 1/10 for HAR pz, with RPA (10%) and 1/10 for HAR pz. with RH (10%). CONCLUSIONS: Resection and primary anastomosis, (correct indications and expert surgeon), gives prevalence of complications and mortality similar to the repeated surgical procedure, a better life quality, but an higher number of post-operating hospitalisation days. PMID- 15754699 TI - A painless treatment for patients undergoing Milligan-Morgan haemorrhoidectomy. AB - BACKGROUND: Postoperative pain associated with Milligan Morgan haemorrhoidectomy (MMH) remains problematic. Most patients complaint a severe pain on defecation and in the 1st postoperative week because of secondary infection and sphincter spasm. We studied the effect of metronidazole, lactulose and glyceryl-trinitrate on pain after MMH. METHOD: Thirty patients were randomly assigned to two groups. The 1st received a treatment with oral laxative (lactulose 66.7%) metronidazole and a topical glyceryl-trinitrate ointment at 0.2% both pre and postoperative; the 2nd received a placebo at the same dosage. The amount of pain in the 1st week and at the time of two defecation, postoperative analgesic requirement and time to return to normal activities were documented. RESULTS: A significant differences in the score pain on 2nd and 3rd day and on the 1st two defecation were observed among group. Analgesic consumption was highest in the placebo group. CONCLUSION: The treatment with lactulose, metronidazole and topic glyceryl trinitrate in patients undergoing MMH, seems to reduce secondary pain and increase patients satisfaction and earlier return to work with improvement of cost and benefit. PMID- 15754700 TI - [Gastric foreign body: bezoars. Apropos of two cases]. AB - OBJECTIVE: The authors report their experience about the treatment of two cases of gastric bezoar, treated in curative mode, the first endoscopically and the second with surgical intervention. SETTING: Operative Unit of General and Thoracic Surgery, Department of General and Emergency Surgery, Organ Transplantation, Policlinico, University of Palermo. INTERVENTION: The patients were submitted to curative treatment, one with endoscopic treatment (mechanical fragmentation of phytobezoar and fragments extraction via-overtube), the second with surgical gastrotomy (stamp trichobezoar). There were no procedure-related complications. RESULTS: The two patients were curative and radically treated. Negative 2 years follow-up. CONCLUSIONS: There is no standardized method for the treatment of gastric bezoars. Endoscopic removal of gastric bezoars after fragmentation and using overtube is effective and safe. Surgical intervention, equally safe, is reserved to huge, stamp, impacted or complicated bezoars. PMID- 15754701 TI - [Aberrant pancreas: a rare cause of epigastralgia]. AB - OBJECTIVE: The Authors report their experience about a case of aberrant pancreas that lead epigastralgia. DESIGN: Report of 1 case and endoscopic treatment (upper endoscopy + EUS + endoscopic polypectomy + endoscopic biopsy of the base. Surgical effectiveness evaluation. SETTING: Section of General and Thoracic Surgery, Department of General Surgery, Emergency and Organ Transplantation, Policlinico "Paolo Giaccone", Palermo. INTERVENTION: After correct and sure diagnosis, the patient was submitted to endoscopic polypectomy with radical and curative intention. RESULTS: Complete recovery. Hematochemical and endoscopic follow-up (1 months) negative. CONCLUSIONS: Diagnosis of aberrant pancreas is very hard and always post-resection. Symptoms are poor and makes the clinical diagnosis extremely difficult, except when the mass attains big dimension (>5 cm). The best treatment of aberrant pancreas is based on correct diagnosis (upper endoscopy + EUS + FNAB) and on a radical treatment that, if the lesion is mucosal, is totally endoscopic (endoscopic polipectomy), without specific risk. PMID- 15754702 TI - [Hemorrhagic adrenal pseudocyst: case report]. AB - INTRODUCTION: Adrenal cysts are usually asymptomatic; they are usually identified occasionally during ultrasound or C.T. scans (incidentaloma). Among adrenal cysts the most common types are epithelial cysts and pseudocysts. Intracystic haemorrhage is one of the possible complications of adrenal pseudocysts. CASE REPORT: We report a case of a young woman with right superior abdominal pain, fever and acute anemia. A C.T. scan showed a 10 cm. mass between the liver and the right kidney. To be sure of the nature of this mass also M.R., urography and C.T.-guided biopsy were carried out. This latter only let us make the final diagnosis of hemorrhagic adrenal pseudocyst. Thereafter, a laparotomic right adrenalectomy was performed, with full recovery of the patient. DISCUSSION: Adrenal cysts may cause differential diagnostic problems with masses of contiguous organs like kidney, liver and gallbladder. For this reason, ultrasound and C.T. scans may not be sufficient and must be completed by M.R., urography and/or C.T.-guided biopsy. Intracystic hamorrhage, spontaneous or post-traumatic, may cause to the patient acute anemia which, as soon as the diagnosis is confirmed, indicates surgery. The operation usually is a laparotomic adrenalectomy, since the laparoscopic approach is not sufficient to control large masses with active bleeding inside. PMID- 15754703 TI - Adult intestinal intussusception: a case report of a rare clinical entity. AB - Intussusception in adult is a rare entity that challenges the surgeon opening a wide range of issues in order to define the etiology and therapeutic strategy. Whether to resect or not the bowel is the main question. The answer can be given only after having seen the site of obstruction and the etiology. Colonic intussusception is best treated by resection. Also small bowel intussusception can require resection if a neoplasm is the cause. Peutz Jeghers can be one of these causes as is seen in the case we report. PMID- 15754705 TI - [Acute PIP joint fractures]. AB - Recent PIP fractures are challenging trauma in terms of diagnosis as well as treatment. It must be remembered that the final outcome will have a considerable impact on the global finger and hand function. Immediate mobilization and rehabilitation are mandatory, and may justify a surgical approach and fixation in selected cases. A good understanding of the fracture type is essential and relies in good part on precise, focused and standardized radiographs. Non-displaced fractures are generally treated conservatively. In the proximal phalanx, the orientation of the fracture line dictates the stability of the fracture. Thus non displaced fractures can occasionally be preventively stabilized, in order to allow early mobilization. Displaced fractures should always be anatomically reduced and surgically fixed. A temporary joint stabilization is optional. In the middle phalanx, one must consider palmar and dorsal fractures differently. Palmar fractures include a distal palmar plate avulsion. The degree of impaction will dictate the stability of the joint towards dorsal subluxation. Dorsal fractures include central slip avulsion of the extensor tendon. An antomical reduction and surgical fixation is mandatory to avoid a progressive boutonniere deformity. Prognosis of all the middle fractures is closely dependent on the degree of impaction. When direct osteosynthesis is not possible, distraction devices, bone graft or palmar plate reconstruction may be useful alternatives. In complex fractures, bone fixation and joint stabilization must be combined in order to prevent secondary displacement and joint instability. PMID- 15754704 TI - [Petit's lumbar hernia: about a case]. AB - The authors are surgeons of Moscati Hospital in Aversa (CE)-Italy, and one of them is a Radiologist in the same hospital; they describe a rare case of lumbar hernia, the Petits hernia and describe the anatomical aspects of parietals hernias. Subsequently they describe the diagnostics aspects and then the surgicals treatments. In this case-report the authors describe the surgical operation to use one prolene-net to close the parietal breach. The most important aspect, that the authors emphsasize, is the absolute importance of careful clinic examination to diagnose it. This work is completed with some pictures that show it; some pictures are made in operatory-room during the surgical treatment and some of them are TC images. PMID- 15754706 TI - [Unreconstructible radial head fracture: resection, implant of Swanson or prosthesis? Retrospective comparative study]. AB - Treatment of unreconstructible comminuted fractures of the radial head remains a therapeutic challenge. Thirty two patients sustained unreconstructible radial head fracture between 1969 and 1999 and have been treated by resection (16 patients), by Swanson implant (8 patients), or by Judet prosthesis (8 patients). The three groups of patients were reviewed clinically and radiologically by two surgeons not involved in their treatment. Functional outcomes of the elbow (Morrey scoring-SOO scoring system, instability, cubitus valgus) and involvement of the wrist (pain, grasp, RUD instability) were evaluated with a mean follow up of 15 years (6-27.7). Elbow or wrist arthritis, ulnar variance, and evolution of Swanson implants were evaluated on standard radiographs. The Judet prosthesis group was evaluated with DASH scoring. Half of the patients were male and 1/3 had a work-related injury. Regarding the following criteria, there was no significant difference between the resection and the Swanson group: Morrey scoring 77/100, SOO scoring 7.4/11, mean flexion was 130 degrees, mean deficient extension was 18 degrees, mean pronation 60 degrees, mean supination 67 degrees, grasp reached 90% of the controlateral side. Arthritis was noted with the same frequency at the elbow (87%), and wrist levels (66%) in each group. Ninety four percent (94%) of patients in the resection group and 89% in the Swanson group were satisfied. Excellent and good results were reported more frequently in the Swanson group (37% resection group, 51% Swanson group). In the resection group the following complications were significantly more frequent: ulnar nerve irritation (2 x), ulnar head dislocation (2 x), ulnar head instability (3 x), para articular ossification (5 x), ulnar variance positive in all cases (mean value 3.20 mm). In the Swanson group only two implants were found to be destroyed at follow-up. In the prosthesis group function and satisfaction were higher than the two other groups but with a shorter follow up. Excellent and good results were reported in 62.5% cases with a DASH scoring between 0 and 16.7. Comminuted fractures of the radial head treated by resection or Swanson implant are both followed by fair results (same functional scoring). Only the level of complications differs between groups: at elbow level for Swanson group, at wrist level for the resection group. Patients in the Swanson group were most frequently satisfied. Metal radial head implant is an attractive solution yielding good functional outcomes in recent reported limited series and in our personal experience, but with a very short follow-up. PMID- 15754707 TI - [Results of the ARPE trapezometacarpal prosthesis: a retrospective study of 37 cases]. AB - We report the outcome of the ARPE trapezometacarpal prosthesis in the treatment of primary osteoarthritis of the thumb. The prosthesis based on a ball and socket system and is uncemented. This study presents the results of 37 consecutive implantations in 29 patients; 28 women, one man, eight bilateral (21 right, 16 left). The mean age at operation was 67 years at a mean of 36 months follow-up (maximum 7 years). Four patients were excluded (1 died, 3 lost to follow up). Preoperative radiological grading according to Comtet was 1 or 2. Patients all had low functional demand. Most of the patients were satisfied and pain free. Mobility was increased, including dorsal extension of the first metacarpal. Radiographic analysis and measurement of the scaphometacarpal index demonstrated a postoperative increased length of the thumb column, with a slight decrease at follow up. This phenomenon is due to sinkage of the prothesis into the cancellous bone. Mechanical complications occurred in four patients within the first four years. Removal of the prosthesis, trapeziectomy and tendon strip plasty were performed in three cases with good final results. This prothesis is a good surgical treatment of primary, isolated TM osteoarthritis in patients with minimal osteoporosis and low functional demands. PMID- 15754708 TI - Intermediate and long-term outcomes following simple decompression of the ulnar nerve at the elbow. AB - INTRODUCTION: There is currently little consensus regarding the appropriate surgical approach to treatment of cubital tunnel syndrome (CubTS), and few studies have reported long-term follow-up of patients who have received surgical treatment for ulnar nerve compression at the elbow. METHOD: Seventy-four patients with a total of 102 cases of CubTS treated with simple decompression of the ulnar nerve were examined 1.0-12.4 years postoperatively. Ulnar nerve conduction studies (slowest conducting 5 cm segment of ulnar nerve motor fibers measured at the elbow) were performed both pre- and postoperatively. The primary clinical outcome was percentage relief of symptoms, divided into "excellent" outcome group or less (> or = 90% improvement or < 90% improvement). RESULTS: Ulnar nerve conduction improved pre- to postoperatively, but clinical improvement was not related to changes in velocity. Women reported greater clinical improvement than men, and weight gain in men (but not women) predicted less improvement. Relief of cubital tunnel symptoms was greatest for those arms receiving carpal tunnel release surgery simultaneous or subsequent to cubital tunnel release. DISCUSSION: Simple decompression may offer excellent intermediate and long-term relief of symptoms associated with CubTS. Although improvement in ulnar motor nerve conduction velocity occurs following treatment of CubTS, it may not be a consistent marker of perceived symptom relief. Finally, these findings suggest that less complete relief of symptoms following ulnar nerve decompression may be related to unrecognized carpal tunnel syndrome or weight gain. PMID- 15754710 TI - [Carpal tunnel syndrome revealed by digital ulcerations caused by arterial vasospasm]. AB - Carpal tunnel syndrome presenting as a trophic disorder in the hand is unusual. We report the case of a 66 year old man in whom nerve conduction tests confirmed compression of the median nerve in the carpal tunnel, and vasospasm in the corresponding area was demonstrated by arteriography for, we believe, the first time. Simple decompression of the carpal tunnel brought about total recovery. The mechanism of the autonomic disorders found in carpal tunnel syndrome remains controversial. The clinical case that we report objectively demonstrates the vasospasm caused by the compression of the autonomic fibres of the median nerve. PMID- 15754709 TI - [Osteoma osteoid of the trapezoid bone: a case-report and review of the literature]. AB - Osteoid osteomas are often localised in long bones but only rarely in the carpus. This is a case report of an osteoid osteoma in the trapezoid. Diagnosis was difficult and established late, due to misleading initial symptoms. Radiological examination allowed localisation of the nidus. For osteoid osteoma localised in a carpal bone, we recommend total removal without bone-graft or arthrodesis. PMID- 15754711 TI - [Melanoma of the forearm. A historical case report]. AB - We present the case of a melanoma of the forearm notable for its depth (Breslow index 35 mm), due to a longstanding lack of interest and care by the patient as well as her physician. Because of systemic metastases, only palliative treatment, consisting of an excision of the lesion for local hygiene, could be offered. The patient survived for only three months. This case illustrates the absolute necessity to remove without delay, any cutaneous tumor that resembles a melanoma, in order to avoid diagnosis being made at a late stage when curative treatment cannot be performed. PMID- 15754712 TI - [Neonatal Volkmann's compartment syndrome. A report of two cases]. AB - Neonatal Volkmann's compartment syndrome is a rare entity. This diagnosis may be suspected when a case presents cutaneous damage associated with poor hand and wrist function after delivery. We present two such cases of neonatal Volkmann compartment syndrome with long term clinical and x-ray follow-up. In our patients, a hand surgeon was not consulted in the perinatal period and early fasciotomy was not performed. No particular aetiology or associated cerebrovascular accident was found. A series of operations was necessary in order to improve function of the hand. Neonatal Volkmann compartment syndrome must be recognised early in order to enable further investigation of any underlying condition and to perform early surgical decompression. Long term clinical and x ray follow-up is necessary to prevent and treat wrist deformation and finger contractures. PMID- 15754713 TI - [An uncommon form of delayed radio-induced brachial plexopathy]. AB - Radiation therapy of the neck or axillary areas for cancer may result in delayed brachial plexus palsy. Differential diagnosis between radiation and tumor brachial plexopathy is difficult. We report the case of a 38-year-old woman, treated by radiation therapy for osteosarcoma of the humeral head 22 years before, who exhibited a rapidly progressive incomplete hand palsy. EMG study revealed a conduction block at the level of the lateral cord. In this case, MRI could not distinguish between a delayed radiation injury and tumor infiltration. The diagnosis was clarified with an ultrasonographic examination. Neurolysis and epineurotomy of the median trunk in the brachial and axillary areas were performed. Histological examination confirmed radiation-induced nerve injury. The characteristics of this uncommon form are reviewed with regard to the previously reported descriptions. PMID- 15754714 TI - Septic monoarthritis of the first carpo-metacarpal joint caused by Mycobacterium kansasii. AB - A case of septic carpal monoarthritis due to Mycobacterium kansasii developing 16 months after accidental inoculation in a healthy laboratory technician is reported. No predisposing factor such as immunosuppression, preexisting degenerative, inflammatory arthritis or cortisone injection was present. Treatment with antituberculous oral medication alone resulted in resolution of the disease. Synovectomy was unnecessary. Ten years after the initial causative event, the patient remains free of symptoms. PMID- 15754716 TI - Tackling the sexual health crisis head on. PMID- 15754717 TI - Where excellence is standard. PMID- 15754718 TI - Developing a Parkinson's Disease service. AB - People with Parkinson's Disease need information and support if they are to maintain an independent lifestyle. The establishment of a dedicated team in Northumbria for these patients is helping them and their carers to become involved in their care by offering education, counselling and information about medication. PMID- 15754719 TI - The nurse's role in promoting weight loss and encouraging healthier lifestyles. AB - The increasing incidence of obesity in the UK has prompted a range of initiatives to raise awareness of the problem. In view of the well-known links between obesity and chronic disease, the focus is now on addressing the issue as part of patient care by advising adults and children on weight management and changes in diet and exercise. PMID- 15754720 TI - Assessment and good technique are key to effective compression therapy. AB - Compression therapy is the standard treatment for patients with uncomplicated venous leg ulcers. It is crucial, however, to undertake a full clinical assessment before initiating treatment, as inappropriate application can be dangerous. The properties and uses of compression are examined alongside its application in clinical practice. PMID- 15754721 TI - Spotting the signs of depression to ensure early treatment and reduce suicide deaths. AB - Many people who have depression and associated disorders never receive treatment and go on to commit suicide. Early and accurate recognition of the symptoms will enable timely intervention and support for individuals, their family and carers. The symptoms of depression are described, together with current treatment options. PMID- 15754722 TI - Determining the space needed to operate a mobile and an overhead patient hoist. AB - Inadequate bed space affects manual handling techniques and the ability to carry out nursing care tasks. Many nurses will join a trust on the basis of the workspace design of the wards. This paper looks at the space required to operate two types of hoist and argues for the recommended bed space dimensions to be increased. PMID- 15754723 TI - Disposable cleansing wipes. AB - With hospital cleanliness becoming a major issue over recent months, a great many campaigns are under way to improve cleaning levels in our hospitals. This article looks at the role of disposable wipes, both for patient skin hygiene and for hard surface cleaning and disinfection, in keeping clinical areas clean and safe for patients. PMID- 15754724 TI - Nicotine replacement. PMID- 15754725 TI - Folic acid: a vital nutrient throughout life. PMID- 15754726 TI - Integrated governance: is this the future? AB - By April 2005, a common framework will be in place for integrated governance throughout the NHS. This poses a challenge for all personnel. Nurses need to be fully involved in planning and leading this process. PMID- 15754727 TI - Don't ignore the carers. PMID- 15754729 TI - Age as a predictor of recurrent hypothyroidism in patients with post-partum thyroid dysfunction. AB - The long term effect of post-partum thyroid dysfunction (PPTD) is not well established. This study was conducted to evaluate the outcome of permanent hypothyroidism and factors predictive of thyroid dysfunction in a large cohort of women with PPTD. In 164 women, treated with levothyroxine for moderate and severe post- partum hypothyroidism, the treatment was discontinued and subjects were followed-up for the recurrence of hypothyroidism. Factors such as thyroid function tests, tyroid peroxidase antibodies (TPOAb) titers, multiparity, history of autoimmune disorders, smoking habits, use of oral contraceptives, family history of thyroid disease, goiter size and age at active phase of PPTD were assessed. Seventy-one post-partum women without PPTD were considered as control group. One hundred forty-eight women (90%) returned for follow-up, and 93 subjects (63%) developed hypothyroidism with a median of 2 months after T4 withdrawal. Of factors predicting recurrence of hypothyroidism, only age, and serum TSH and T3 concentrations at the time of occurrence of PPTD entered the model in multiple stepwise regression analysis. Hypothyroidism occurred earlier and more frequently in women aged 30 and over, as compared to younger ones (p < 0.01), hazard ratio 1.69. In 55 euthyroid subjects with history of PPTD, serum TSH was significantly increased as compared to the control group (3.0 +/- 1.6 vs 0.8 +/- 0.5 mU/l, p < 0.001). These data, representing the largest cohort of women with PPTD who have been followed, show that a significant proportion of women with moderate and severe PPTD, in particular those aged 30 yr and over, develop recurrent hypothyroidism. Proper follow-up of all women with PPTD, the older ones in particular, is recommended. PMID- 15754728 TI - Boxing as a sport activity associated with isolated GH deficiency. AB - Traumatic brain injury (TBI) has long been known as a cause of hypopituitarism, and it is characterized by a high prevalence of neuroendocrine abnormalities. Boxing, one of the most common combative sports, may also result in TBI. As far as we know, pituitary functions including GH status have not been investigated in boxers. Therefore, in this preliminary study, we have assessed the pituitary functions in boxers. Eleven actively competing or retired male boxers with a mean age of 38.0 +/- 3.6 yr and 7 age-, sex- and BMI-matched healthy non-boxing controls were included in the study. Biochemical and basal hormonal parameters including IGF-I levels were measured. To assess GH secretory status in boxers and healthy controls, GHRH (1 microg/kg)+GHRP-6 (1 microg/kg) test was performed. After GHRH+GHRP-6 test, mean peak GH level in boxers and in controls were 10.9 +/ 1.7 and 41.4 +/- 6.7 microg/l, respectively (p < 0.05). Peak GH levels in 5 (45%) boxers were found to be lower than 10 microg/l and considered as severe GH deficient. In the control group, mean IGF-1 levels (367 +/- 18.8 ng/ml) were significantly higher than that obtained in boxers (237 +/- 23.3 ng/dl) (p < 0.01). All the other pituitary hormones were normal including ADH as no signs and symptoms of diabetes insipidus. There was a significant negative correlation between peak GH levels and boxing duration, and between peak GH levels and number of bouts. In conclusion, we think that boxing is a cause of TBI, and GH deficiency is very common among boxers. Further studies including large number of boxers, both professional and amateur, are needed to clarify pituitary dysfunction in boxers. PMID- 15754730 TI - Human resistin gene polymorphism is associated with visceral obesity and fasting and oral glucose stimulated C-peptide in the Quebec Family Study. AB - Obesity and insulin resistance are common features of Type 2 Diabetes. A new protein called resistin has been shown to be secreted by adipocytes in mice and to influence insulin sensitivity. The goal of the present study was to investigate the associations between one polymorphism (g-420C>G) of the human resistin gene and phenotypes related to adiposity and glucose metabolism. We genotyped 725 (including 42 diabetics) adult subjects participating in the Quebec Family Study (QFS) by a minisequencing method. Forty-two were diabetic subjects. Phenotypes measured were: body mass index (BMI) and waist circumference (WC), % body fat (PFAT) and fat mass (FM) assessed by under water weighing, abdominal total, subcutaneous and visceral fat assessed by computed tomography and fasting plasma glucose, insulin and C-peptide and their responses to an oral glucose tolerance test (OGTT). Comparisons between genotypes were performed in non diabetic men (no.=280) and women (no.=403) separately by analyses of covariance (ANCOVA). Among men, g-420 G homozygotes had less visceral fat (p < 0.05), lower levels of acute insulin responses to an OGTT and lower levels of C-peptide in a fasting state and in responses to an OGTT than carriers of the C allele (p < 0.01). These associations were independent of age and adiposity but were not observed in women. These results suggest that in men, the human resistin gene is associated with reduced amount of visceral obesity and lower insulin secretory responses to a glucose load. PMID- 15754731 TI - Sodium iodide symporter expression and radioiodine distribution in extrathyroidal tissues. AB - The functional role of the sodium iodide symporter (NIS) in extrathyroidal tissues was investigated by examining its mRNA and protein expression, together with the evidence of radioiodine (131)I uptake in 302 patients who underwent (131)I total body scanning, following the administration of high doses of (131)I for a papillary or follicular thyroid carcinoma. By using a real-time kinetic quantitative RT-PCR and immunohistochemistry, the expression of NIS protein was detected mainly in secretory tissues. In parallel, 1311 uptake was evidenced in the majority of patients in the salivary glands (in 39%) and stomach (in 78%), but was found in breast in only 4 young female patients. These data demonstrate a strong correlation between the organ radioactivity distribution, as observed in vivo, and NIS protein expression. Interestingly, (131)I is rarely concentrated by mammary glands, even when large doses are administered. Moreover, a (131)I transfer in secretion fluids may represent a potential source of contamination responsible for false positive images and diagnostic pitfalls. PMID- 15754732 TI - Novel somatic MEN1 gene alterations in sporadic primary hyperparathyroidism and correlation with clinical characteristics. AB - Primary hyperparathyroidism (pHPT) is a common endocrine disease that in more than 95% of cases is sporadic and only in some cases is caused by inherited disorders, isolated or as part of multiple endocrine neoplasia (MEN1 and 2). Somatic mutations of MEN1 gene have also been described in sporadic parathyroid tumors. In our study, we examined the presence of alterations in MEN1 gene in a series of 39 patients who had undergone surgery for sporadic pHPT (35 with parathyroid adenoma or hyperplasia, 4 with a carcinoma). A genotype-phenotype correlation was also analysed. After DNA extraction from paraffin-embedded tissues, we amplified by PCR and sequenced the exons 2-10 of the MEN1 gene. Somatic MEN1 mutations were detected in 6 of the 35 patients with a benign parathyroid lesion examined (17.1%), whereas no alterations were found in the carcinomas. Four novel MEN1 gene mutations were identified as follows: one frameshift mutation (222insT, exon 2), one frameshift deletion (912delTA, exon 5), one in-frame deletion (835del18, exon 4) and one missense mutation (P291A, exon 6). In addition, one missense mutation (L89R, exon 2) and one nonsense mutation (Q536X, exon 10) were previously reported. Moreover, two polymorphisms were also found: one allele carried a R171Q polymorphism (1/39 tumors), while a D418D polymorphism (GAC/GAT) was found in 15 and 8 tumors in hetero (CT) and homozygosity (TT), respectively. In no case (mutations and/or polymorphisms) did we find a genotype-phenotype correlation. In conclusion, our data demonstrate the presence of somatic alterations of the MEN1 tumor suppressor gene in about one fifth of benign sporadic parathyroid tumors. The absence of a genotype-phenotype correlation, however, suggests the involvement of other genetic/epigenetic factors for the full expression of the disease. PMID- 15754734 TI - Increased abnormal pituitary findings on magnetic resonance in patients with male idiopathic hypogonadotrophic hypogonadism. AB - Idiopathic hypogonadotropic hypogonadism (IHH) is a well-known disorder apart from its pathogenesis, which is still mostly unclear, even though a diverse subgroup of patients with hypogonadotropic hypogonadism and hyposmia/anosmia--the Kallman syndrome--have been partly linked to a mutated gene, known as kal-1 gene. In this study, we aimed to evaluate the sellar region of patients with IHH on magnetic resonance (MR). Pituitary MR of 120 male patients with IHH, diagnosed by a thorough endocrinologic assessment, were compared with pituitary MR of 49 healthy cases selected randomly who underwent detailed endocrinologic and neurologic evaluation and were assessed as healthy. Patients with IHH were diagnosed with microadenomas and irregularly contrasting pituitary (ICP), 18.2 and 10.7%, respectively. Although some anatomic variations were seen in healthy controls, microadenomas and ICP had solely been observed in patients with IHH and none in controls. Intact appearence of hypophysis in patients with IHH was significantly lower than in randomly selected healthy male subjects (p = 0.021). Mean infundibulum width of hypophysis and transverse diameter of posterior hypophysis were significantly broader in patients with IHH than in controls (both having p < 0.001), while mean hypophysis height and volume did not differ between groups. Results showed unusual incidence of pituitary abnormalities on pituitary MR in male patients with IHH. In conclusion, MR imaging is particularly useful in defining the morphological aspects of the hypothalamo - pituitary region in some endocrine disorders and other researchers might want to bear our findings in mind when performing MR evaluation of similar patient subgroups. PMID- 15754733 TI - Identification of IgG-immunocomplex macroprolactin with an immunometric "sandwich" system: technical and clinical considerations. AB - MacroPRL can be due either to anti-PRL autoantibodies IgG (IgG-macroPRL) or other PRL containing molecules without IgG-PRL immuncomplexes (nIgG-macroPRL), the composition of which is not fully clarified. The aims of this study were: a) testing a new immunometric assay, capable of recognising IgG-macroPRL, b) looking for any biological and/or clinical discrepancy between nlgG-macroPRL and IgG macroPRL. Clinical, biological and neuroradiological data were recorded from 28 hyperprolactinemic women classified as macroprolactinemic on the basis of PRL recoveries after polyethylene glycol (PEG) precipitation <50% on a Roche Elecsys analyser. Fourteen sera with recoveries >70% were employed as controls for the IgG-macroPRL assay. An immunometric "sandwich" assay for IgG-macroPRL was performed employing an anti-PRL antibody to capture PRL and a second one binding and tracing the immunoglobulin G (IgG) component. For this purpose, 2 kits were simultaneously employed, the first for PRL assay and the second for Anti Toxoplasma IgG assay. The procedure was applied to both DiaSorin LIAISON and Abbott AxSYM systems. IgG-macroPRL immunometric detection was obtained with DiaSorin LIAISON. Twenty out of 28 (72%) sera gave luminescent signal higher than the maximum control serum and were considered as IgG-macroPRL carriers. Our results confirm the high IgG-macroPRL prevalence in macroprolactinemia. No significant clinical differences appear between IgG and nIgG-macroPRL. Conversely, significantly higher PRL values (p =0,039) and lower recoveries (p = 0.001) were found in IgG-macroPRL. Further studies with reference methods, such as gel chromatography and human IgG (h-IgG) affinity columns, are needed to fully validate this IgG-macroPRL immunometric assay. PMID- 15754735 TI - Cost-of-illness study in acromegalic patients in Italy. AB - INTRODUCTION: acromegalic therapeutic goals are directed at removing the tumor, preventing tumor re-growth and reducing long-term morbidity and mortality. In this scenario, the acromegalic patient needs a variety of health resources (diagnostic tests, surgery, radiotherapy, specialist visits and drugs) for his/her cure, in order to decrease/stop the progression of the disease and to cure the co-morbid diseases. Lack of epidemiological data has suggested performing an Italian retrospective study aiming to assess the health resource consumption that is caused by acromegalic cure and the relative co-morbidities, in order to estimate the amount of the direct costs of acromegalic patients. METHOD: a retrospective study was performed on a total of 134 patients (142 patients selected, 76 in Genoa and 66 in Turin) for a period of about 7 yr preceding the enrolment date. Only direct costs were evaluated by performing an analysis on the perspective of Italian Healthcare Service (SSN). RESULTS: the mean total direct costs for acromegaly cure ranged from 7,968.41 to 12,533.02 Euros/yr (p < 0.01; Mann Whitney Test), respectively, for Responders and Non Responders. The cost driver was drug (SS analogs) for acromegalic cure. The co morbidity conditions associated to acromegalic Non-Responder patients are clearly higher than those with well-controlled disease. CONCLUSION: the study supports the hypothesis that controlled patients drove a saving for SSN in comparison to poor control patients that use more health resources. PMID- 15754736 TI - Treatment of acromegaly with SS analogues: should GH and IGF-I target levels be lowered to assert a tight control of the disease? AB - BACKGROUND: in acromegaly, the criteria for the cure of the disease after neurosurgery have become tighter and tighter. In contrast, the evaluation of control of disease activity during medical treatment is based upon the normalisation of IGF-I levels and epidemiological criteria, i.e. lessening GH (assessed by RIA) to levels reported to normalise increased mortality. The aim of this study was to evaluate GH and IGF-I suppression during prolonged SS analogues (SA) treatment. The concordance between "safe" GH and normalised IGF-I levels during SA was also assessed, according to gender and gonadal status. DESIGN: multicentre, retrospective. Patients. GH/IGF-I levels were evaluated in 207 acromegalic patients (132 females, aged 20-85 yr) during a prolonged treatment (longer than 12 months) with individually tailored doses of depot SA( lanreotide or octreotide-LAR in 97 and 110 patients, respectively). Final IGF-I levels were transformed in z-scores using data collected in a large cohort of normal subjects of 3 different age groups (20-40 yr old, 41-60 yr old, 61-80 yr old, n=160, 148, 115, respectively), that allowed to set up quartiles of normality (I = 3rd-25th percentile, II = 26th-50th, III = 51th-75th, IV = 76th-97th). RESULTS: fifty-nine and 19.3% of patients achieved GH levels <2.5 and <1 microg/l, respectively. IGF I were normalised (z-score between 2 and -2) in 58.4% of patients. The distribution of normal IGF-I values among quartiles was uneven: 7%, 19%, 25%, and 49% of values were distributed in the I, II, III, and IV quartile, respectively. The concordance between GH and IGF-I values was poor: 28.4% of patients attaining GH values <2.5 microg/l had still pathological IGF-I (even 12.5% of those with GH <1 microg/l), and 39.3% of those with GH levels still above the "safe" limit had "nor IGF-I. Although proportions of IGF-I normalisation were not different between males and females, the regression line obtained between GH and IGF-I z score showed the same slope but with a significantly lower intercept in regularly cycling women than in males and in postmenopausal females. Thus for any GH value, cycling females had lower IGF-I than menopausal women and males, and their IGF-I normalisation could be achieved by higher GH values. By ROC analysis, the achievement of normal IGF-I was predicted by the GH value of 1.8 microg/l in males and 2.4 microg/l in females. CONCLUSIONS: in acromegalic patients on SA treatment, GH and IGF-I levels are often not concordant. In addition to age, sex is to be taken into account in the evaluation of hormonal targets. A better refinement of GH and IGF-I targets to be reached while on treatment with SA is warranted. PMID- 15754737 TI - Hypopituitarism as a consequence of traumatic brain injury (TBI) and its possible relation with cognitive disabilities and mental distress. AB - Recent studies have demonstrated that hypopituitarism, in particular GH deficiency, is common among survivors of traumatic brain injury (TBI) tested several months or yr following head trauma. We present the results of endocrine, neurological, neuropsychological and psychiatric evaluation in a group of 67 patients who suffered TBI at least one yr ago. Our study shows that decreased endocrine function is either restricted to one or more anterior pituitary hormones and is present in 34% of patients with any pituitary hormone deficit, while multiple pituitary hormone deficiencies are found in 10% of patients. GH/IGF-I axis was evaluated by GHRH+GHRP-6 test and IGF-I measurement. Severe GHD is the most frequent deficiency present in 15% of TBI patients. Gonadotrophin deficiency was present in 9% of patients with TBI, while thyrotroph and corticotroph function seemed more refractory to impairment. Patients with moderate-to-severe trauma are not necessarily more likely to have hypopituitarism than those with mild injury. Neuropsychological testing revealed a significant positive correlation of peak GH levels after GHRH+GHJRP-6 test with verbal learning and verbal short term memory (RAVLT total score p = 0.06, immediate free recall p = 0.02 and delayed free recall p = 0.04). Verbal and visual memory was significantly lower in elderly patients and in males. Visoconstructional abilities (RCF copy) were significantly lower in the elderly (p < 0.01) and undereducated (p = 0.02). Visual memory (free recall of complex figure after 30 min) significantly correlated with lower IGF-I levels (p = 0.01). Gonadotrophins and testosterone correlated significantly with visoconstructional abilities. Simple and complex conceptual tracking (TMT A and B) was significantly more impaired in older TBI patients (p < 0.01) and with longer time from trauma (TMT B only, p = 0.03). The psychiatric evaluation by using two different scales showed depression, phobic anxiety and psychoticism to be more prominent in the TBI group. Paranoid ideation and somatization negatively correlated with the peak GH responses to GHRH+GHRP-6 test (p = 0.04 and p = 0.03, respectively). Depression scale showed that nearly half of patients suffered from mild to moderate depression. The benefits of hormone replacement therapy on cognitive functioning and mental distress in TBI patients are eagerly awaited. PMID- 15754738 TI - Effect of cabergoline treatment on Cushing's disease caused by aberrant adrenocorticotropin-secreting macroadenoma. AB - The present case involves a 47-yr-old woman with Cushing's disease due to pituitary macroadenoma. The patient had suffered from hypertension and obesity for two yr. Her serum cortisol levels were moderately elevated throughout the observation period, and dexamethasone failed to suppress the cortisol secretion. Plasma ACTH levels were markedly high (>100 pg/ml) and did not respond to CRH provocation. Gel filtration analysis of the patient's plasma detected the existence of big ACTH molecules, which eluted with a peak of authentic 1-39 ACTH. Cranial magnetic resonance imaging (MRI) revealed a 3 cm pituitary tumor occupying the sellar region and right cavernous sinus with diffuse enhancement by gadolinium. The pituitary mass was removed by transsphenoidal surgery, and was pathologically identified as compatible to ACTH-producing pituitary adenoma by immunohistochemistry. RT-PCR analysis of total cellular RNA extracted from the resected adenoma revealed a relatively high expression level of dopamine D2 receptor (D2R) mRNA. Therefore, a long-acting D2R agonist, cabergoline (0.25 to 0.5 mg/week), was administered for the remnant adenoma, which gradually reduced ACTH levels in 90 days. In addition, cranial MRI exhibited shrinkage of the remnant pituitary mass after a 6-month treatment with cabergoline. This case demonstrates the efficacy of cabergoline to treat Cushing's disease caused by pituitary macroadenoma secreting aberrant ACTH molecules. PMID- 15754739 TI - A rare case of adulthood-onset growth hormone deficiency presenting as sporadic, symptomatic hypoglycemia. AB - Symptomatic hypoglycemia is described in children with severe GH deficiency (GHD), but is rare in adults with GHD. We describe the case of a 62- yr-old man, referred for recurrent hypoglycemic events. He reported a previous head trauma at the age of 20 yr and a diagnosis of reactive hypoglycemia at the age of 50 yr. In the last months, during a period of job-related stress, the hypoglycemic episodes became more frequent and severe (glucose <2.2 mmol/l), finally requiring hospitalization. On admission, the patient was in good general health, with normal renal and hepatic function. During hospitalization, no hypoglycemic episodes were recorded, also during a 72-h fasting test. Biochemical data and abdominal computed tomography (CT) excluded insulinoma. A tumor-induced hypoglycemia was ruled out. The 4-h oral glucose tolerance test (OGTT) showed an impaired glucose tolerance with a tendency toward asymptomatic hypoglycemia. Hormonal study disclosed low levels of GH (0.2 ng/ml) and IGF-I (51 ng/ml); the response of GH to GHRH plus arginine confirmed a severe GHD (GH peak 2.7 ng/ml). Other pituitary and counterregulation hormones were within the normal range and magnetic resonance imaging (MRI) of the pituitary gland was normal. Replacement therapy with a low dose of rhGH induced an increase of IGF-I up to low-normal values, accompanied by lasting regression of hypoglycemic events. In conclusion, hypoglycemia was the main clinical symptom of isolated adult onset GHD, in the present case. The possible pathogenesis of isolated adult onset GHD and the association of GHD with conditions predisposing to hypoglycemia are considered and discussed. PMID- 15754741 TI - Turner syndrome and GH treatment: the state of the art. PMID- 15754740 TI - Thyroid, hemostasis and thrombosis. AB - The aim of this paper is to briefly review some practical aspects of the relationship between thyroid function and several disorders of the hemostatic system in terms of bleeding and thrombosis. Thrombocytopenia, acquired hemophilia, hypercoagulability, cardioembolism and other biochemical coagulative and fibrinolytic abnormalities have been described in the past years both in hyper- and hypothyroidism. Since most of hyper- and hypothyroid conditions are the consequence of autoimmune thyroid disease (1), either deranged immune function, altered circulating thyroid hormone concentration, or both may concur in the pathogenesis of hemostatic disorders of potential crucial clinical impact. These aspects will be outlined and discussed in an attempt to give answers to some questions, often arising in the clinical approach. PMID- 15754742 TI - Missense mutations in the human insulin promoter factor-1 gene are not a common cause of type 2 diabetes mellitus in Taiwan. AB - Type 2 diabetes mellitus (T2DM) is a common metabolic disorder characterized by a hyperglycemia resulting from defect in insulin secretion and insulin action. Recent studies showed that dominant negative mutations in the insulin promoter factor-1 (IPF-1), a pancreatic beta-cell specific transcription factor, cause maturity-onset diabetes of the young (MODY), a subtype of T2DM with early onset and monogenic autosomal inheritance. In addition to MODY, IPF-1 mutations are suggested to predispose to common late-onset T2DM with different penetration of the mutations reflected in their in vitro activity. Thus, we investigated IPF-1 C18R, Q59L, D76N and R197H mutations in Taiwanese patients with common late-onset T2DM, because research into IPF-1 variants in Taiwanese diabetic patients--a population with the lowest range of diabetic incidence--has never been documented. Peripheral blood samples were collected and genomic DNA was extracted from 434 patients with T2DM and 194 non-diabetes control study subjects. IPF-1 genetic variations were analyzed by PCR and restriction fragment length polymorphism (RFLP) analysis. We did not find any of these four IPF-1 mutations in our patients. Our results suggested that IPF-1 mutations were not a common cause associated with Taiwanese T2DM. PMID- 15754743 TI - Michelangelo, Goiter and...cats. PMID- 15754744 TI - [Use of a preterm formula with long chain polyunsaturated fatty acids: acceptability and tolerance]. AB - INTRODUCTION: The majority of preterm formulas have included long chain polyunsaturated fatty acids (LCPUFAS) for the relationship with visual and cognoscitive brain development. This work tried to probe the tolerance and acceptability of a preterm formula from the point of view of palatability for the LCPUFAs origin. METHODS: Infants under 37 weeks of gestational age at birth and birth weight below 2,500g with the possibility of being fed by suction and not receiving breast milk were included. Using a scale modified from "Pain Analog Scale," the nurse who was feeding the infant had to qualify the baby's facial expression at these moments. RESULTS: 50 infants with age 37.2 +/- days of life at least receiving the formula for 3 complete days were included. Ninety percent of them had weight gain each day and the third day was the most. There were no problems in number and frequency of stools. Thus, 782 evaluations by the nurses were made (15 for each patient); 66.8% of the time the infants accepted the formula with no problem; 21.1% were satisfactory; 3.7% were very satisfactory. Also, 91.8% of the nurses declared their satisfaction with the formula, and in 4 cases they declared dissatisfaction. They attributed it to the patient's pathology. PMID- 15754745 TI - [Prevalence of non-alcoholic esteatohepatitis in adults with metabolic syndrome in Oaxaca]. AB - Nonalcoholic steatohepatitis (NASH) is a chronic liver disease that occurs in patients with no significant alcohol consumption, characterized for macrovesicular steatosis, hepatocellular necrosis, mixed inflammatory infiltrate and various grades of fibrosis and, in some cases, Mallory bodies. The prevalence of this disease is unknown; recent studies indicate that in the general population, incidence is about 3%, although the rates are higher in some subpopulations as are obesity and diabetes mellitus. The goal of this work was to determine the prevalence of non-alcoholic steatohepatitis in one sample of patients with the metabolic syndrome, patients at "Presidente Juarez" Regional Hospital. It was a cross-sectional and descriptive study in which, by means of random selection a sample of 110 patients with type 2 diabetes mellitus, arterial hypertension, obesity and dyslipidemia, alone or in combination was chosen. We carried out a clinical and laboratory evaluation, selecting those patients with persistent elevation of aminotransferases for whom additional studies were made, in order to exclude other causes of enzyme abnormality. Five patients were selected for percutaneous liver biopsy, all they had NASH. The prevalence of NASH was 4.54%. The clinical and laboratory characteristics of these patients will be discussed in the article. PMID- 15754746 TI - [Toxicity of venoms from snakes of medical importance in Mexico]. AB - The characterization of the toxic activities of snake venoms is necessary to understand the physiopathology of the envenomation and to test the potency of the antivenoms used to treat this pathology. Because of the lack of data on the toxic activities of venoms from Mexican snakes of medical importance, we studied the venoms from Bothrops asper, Athropoides nummifrr, Agkistrodon billineatus, Crotalus durissus durissus, Crotalus basiliscus, Crotalus scutulatus, Crotalus atrox and Micrurus nigrocinctus. The studies performed were: SDS-PAOE, determination of lethal potency, hemorrhagic, necrotizing, coagulation on plasma and fibrinogen, phospholipasic and fibri(noge)nolytic activities. In addition we studied the neutralizing capacity of the toxic activities of an antivenom currently used for the treatment of snakebites in Mexico. The venom from viperids showed important hemorrhagic, necrotizing, coagulative on plasma, prothrombinic, fibrinogenolytic and phospholipase activities. The venoms with the highest lethal potency were those of Micrurus nigrocinctus and Crotalus scutulatus; however, the viperine venom that globally displayed the most potent toxic activities was from Bothrops asper. All the venoms showed toxic activities of similar range to those described for other American venomous snakes. The activity on plasma or fibrinogen varied widely among the different venoms but all displayed capacity to act on the coagulation system. The antivenom tested not only neutralized the lethality B. asper venom but also its other toxic activities. PMID- 15754747 TI - [Lichen planus and hepatitis C virus. Is there an association?]. AB - INTRODUCTION: The etiology of lichen planus (LP) is unknown. The possible role of hepatitis C virus (HCV) has been studied with controversial results. MATERIALS AND METHODS: The aim of this study was to determine the prevalence of HCV infection in 36 patients with lichen planus (LP) confirmed by histopathology, 12 in a retrospective way, and 24 in a prospective one. The controls were 60 consecutive blood donors from the Blood Bank of the hospital. Third generation enzyme immunoassay screening test was employed for determination of antibodies against HCV. RESULTS: A prevalence of HCV infection of 2.77% in LP patients, and 0% in control group (Fisher's exact test--p=0.375). CONCLUSIONS: We found in our patients with LP a lower prevalence of positive serology for HCV than that reported in the literature. Since all studies have found patients with LP without infection by HCV, we think that other factors in the pathogenesis of this dermatosis need to be studied. PMID- 15754748 TI - [Laser therapy in orthognatic surgery]. AB - This study describes the prevalence of patients with dentofacial deformities attended in the Maxillofacial Surgery Department of Hospital de Especialidades del Centro Medico Nacional Siglo XXI of the IMSS, during January to December 2002. Twenty-six patients received postoperative laser therapy in the right area of the mandible ramus, after bilateral sagittal modified osteotomy (BSMKSO). The results with laser therapy showed less pain and edema in comparison other patients without this therapy. The results were carried out after 24 hours, 3 days, 8 days and 15 days, and after 10 treatments of laser therapy. The X-Ray and biopsy showed bone healing 30 days, six and eight weeks after orthognatic surgery procedures. PMID- 15754749 TI - [CD1 pathway and NK T cell activation to glycolipid antigens from Mycobacterium tuberculosis]. AB - The aim of this review is to analyze the current state of our knowledge about cell surface molecules involved in glycolipid antigen presentation, named CD1 family. These proteins constitute a third class of antigen-presenting molecules. CD1 molecules develop diverse important immune functions in host defenses against microbial infections. In recent years these proteins have been involved in the generation of cell-mediated immune response against Mycobacterium tuberculosis. Here, we analyze relevant roles of CD1 proteins and glycolipid antigen-specific T cells. PMID- 15754750 TI - [A 66 year-old male with a submandibular mass, fever and productive cough]. PMID- 15754751 TI - [Clinical evolution in two patients with Wiskott-Aldrich Syndrome]. PMID- 15754752 TI - [Kikuchi-Fujimoto disease in pediatric population: a case report and a review of literature]. AB - A 10 year-old child, with a history of a right cervical mass, is admitted to the Costa Rican National Children's Hospital for workup. The mass appeared approximately 4 weeks before admission. Laboratory tests were performed and malignity, infection and immunologic causes were ruled out. A biopsy was performed revealing granulomas characterized by central necrosis with abundant karyorrhexis, surrounded by histiocytes, lymphocytes and giant multinucleated cells, without neutrophils. Special stains showed no microorganisms. Once infectious and immunologic causes were excluded, and based on the biopsy's result, treatment was ruled out. Twelve months later, the patient is still asymptomatic; therefore, the diagnosis of a Kikuchi-Fujimoto syndrome was proposed. This report constitutes the first pediatric case diagnosed in our country. PMID- 15754753 TI - [Hospitals' evolution through the ages]. AB - The predecessor institutions of modern hospitals--Byzantine nosocomeion, European hospitale and Islamic maristan--were dissimilar both in their patients and their aims. The first charitable organizations in West Europe (Rome) and in the East (Cesarea in Cappadocia) were rather hospices. After the collapse of the Western Roman Empire (476 A.D.), some monastic centers were prepared to provide medical assistance to religious and secular patients. Since the XI and XII Centuries in all of Christian Europe the charitable institutions, designated as hospitale, multiplied. Among the Italian ones, the Roman Santo Spirito (Holy Ghost) Hospital, built in the 1201-1204 period, reached a preeminet position. This one soon became the most important of the entire Christendom (archihospital), with a lot of affiliated hospitals in Europe and later in America. The first American hospital, Saint Nicholas Hospital, opened on December 29, 1503 in Santo Domingo, obtained in 1541 its affiliation to the Santo Spirito archihospital. Regarding continental America, the first health centers were established in Mexico: the Immaculate Conception Hospital and the Saint Lazarus Hospital, both established by Hernan Cortes. For its part, clinical teaching was systematized at the Saint Francis Hospital in Padua and by there moved to Leyden. In Mexico, the chair of medical clinics or practical medicine was established in 1806 at the Saint Andrew Hospital. During the XX century, Dr. Ignacio Chavez was the driving force behind the creation of the modern Mexican Health Institutes. These ones are dedicated to the treatment of poor patients, as well as to medical teaching and research. PMID- 15754754 TI - [Secondary non-Hodgkin's B-cell lymphoma involving the breast: radiologic imaging]. AB - Secondary lymphomas involving the breast, although uncommon, represent the largest group of metastasic tumors to the breast. Primary non-Hodgkin lymphomas (NHLs) of the breast are also rare, accounting for 1.7% to 2.2% of extranodal NHL cases and 0.38 to 0.7% of all NHLs. Approximately 300 cases have been reported in the Medical literature. Mammographic appearances are described as round or oval areas of opacity. The breast mass may appear homogeneous or inhomogeneous. The lesion may be very well defined and may be mistaken for a benign process, most notable in patients younger than 35 years of age. Ultrasonographic appearance is described as a sharply defined mass with low or medium echoes. The posterior aspect of the mass is well defined (possibly with slight acoustic enhancement but with no associated posterior shadowing). In summary, the mammographic appearance of the breast lymphoma is nonspecific, but the diagnosis can possibly be excluded if calcifications or a desmoplastic reaction are present. Prominent lymph vessels in a patient with a breast mass should raise the suspicion of breast lymphoma. The most critical aspect in the workup of a breast mass is the tissue biopsy, since radiographically lymphomatous lesions are indeterminate. We present the case of a 79-year-old woman with abdominal pain and a palpable breast mass; we also make a brief summary of the clinical features and main imaging findings of NHL (plain radiograph, mammography, breast ultrasound and thorax tomography). PMID- 15754755 TI - [Molecular diagnosis in patients and carriers of hemophilia A and B]. PMID- 15754756 TI - [The 20th century: 100 years of misfortune and splendor]. AB - The 20th century has been one of the most intense and convulsive periods in the History of humanity. A century of paradoxes and contrasts, it began with optimism, it witnessed the apocalypse of two world wars, and finished with unimaginable scientific progress that gave us a new civilization that we cannot yet grasp. In this century, significant events happened that shaped our time and projected their results toward an immediate future. Some of these were providential in understanding man's life, fighting against illnesses and prolonging life, and others were of undeniable social importance for humanity. Some knowledge was based on the work of others. Philosophy was embedded in mathematics, as was science in philosophy, while politics and the economy exercised so decisive an influence in our way of feeling and living that culture and society were affected to the core. Within that century the biggest technological revolution of all the time was also created, as transcendent as it was unimaginable, which put mankind on the road to the stars with the moon landing and in the process created the information society whose signature symbol, the internet, emerged as a new demiurge. However, the 20th century, with all its misfortune and splendor, paradoxes and contrasts, creation and destruction, was the most transcendent in the whole of history and it bequeaths to the future a promising horizon in the search for a renovated meaning of life and a yearning for peaceful coexistence for the whole humanity. PMID- 15754757 TI - [Lifestyle-related diseases: Hypertension]. PMID- 15754758 TI - [Lifestyle-related diseases: Dyslipidemia]. PMID- 15754759 TI - [Renal transplantation and the strategy of treatment for pediatric patients with chronic renal failure]. PMID- 15754760 TI - [Recent review of renal allograft pathology]. PMID- 15754761 TI - [Extracellular-superoxide dismutase production in mesangial cell growing in extracellular matrix]. AB - To study the protective function against oxygen radicals in the mesangial area, we assessed extracellular superoxide dismutase (EC-SOD) production in mesangial cells (MCs) in vitro. These cells have a major protective function against oxygen radicals in the extracellular space. In two different kinds of culture conditions: "growth medium" with fetal cow serum, and "differentiation medium" with reduced growth factor, and four extracellular matrixes; type I collagen, type IV collagen, laminin and fibronectin, were added to the MC culture. With the difference in the culture media, differentiation medium induced EC-SOD hyper production associated with the both of the slowing down of cell proliferation and the suppression of IL-6 and IL-8 production. With difference in the extracellular matrix, the presence of type VI collagen and laminin promoted higher production of EC-SOD than fibronectin and type I collagen. Type IV collagen and laminin associated with the physiological condition of the glomeruli promoted EC-SOD production compared with the presence of type I collagen and fibronectin dominantly located in pathological condition. Suppression of EC-SOD production in growth medium along with MC proliferation and chemokine hyper-production compared with production in differentiation medium might mimic reduction of the protective capacity against oxygen radical toxity during mesangial proliferation in the glomerular nephritis. MC proliferation with type I collagen and fibronectin might enhance oxygen radical toxity in the glomeruli, and accelerate glomerular sclerosis through the suppression of EC-SOD production. PMID- 15754762 TI - [Antihypertensive therapy for refractory morning hypertension in patients on peritoneal dialysis]. AB - Due to excessive salt and water retention, hypertension often becomes refractory in patients undergoing peritoneal dialysis (PD). Management of high blood pressure (BP) appears to be of particular importance in such patients because of its substantial impact on the patients' prognosis. However, attempts to control hypertension in PD patients have not been successful in most cases. In this regard, the present study aimed to address the adequacy of current antihypertensive therapy for PD patients. A new antihypertensive strategy expected to improve the outcome was tested on the assumption that treatment with either angiotensin converting enzyme inhibitor (ACE-I) or angiotensin receptor blocker (ARB) in the evening together with alpha1-blocker at bed time and long acting Ca channel blocker (CCB) in the morning might ameliorate BP control associated with morning hypertension. Enrolled in the present study were 40 patients whose BP was evaluated by both office and home measurement. Due to an emerging concern about morning hypertension, home BP measured early in the morning was used for the analysis. Each patient was categorized into the following four groups in accordance with office and home BP: well-controlled, poorly-controlled, white-coat and masked (opposite to white-coat), hypertension. After the observation period, 28 patients with refractory hypertension were allocated to intensive antihypertensive therapy in which ARB or ACE-I previously prescribed in the morning or daytime was shifted to the night. In addition, alpha1-blocker was given at bed time. Furthermore, long-acting CCB and diuretics were shifted to the morning. The patients were then followed up for 4-6 months. The results were as follows: 1) Of the total number of 40 PD patients, systolic hypertension was noted in 50% of cases by office BP and in 80% by home BP. The former was less frequent than the latter (p=0.0047, n=40). Similarly, diastolic hypertension was noted in 20% by office BP and in 45% by home BP. The former was less frequent than the latter (p=0.0045, n=40 by McNemar's analysis). The distribution of BP control categories was well-controlled in 11%, poorly controlled in 42%, masked hypertension in 39% and white-coat hypertension in 8% when determined by systolic BP. The distribution was well-controlled in 45%, poorly-controlled in 13%, masked hypertension in 34% and white-coat hypertension in 8% of cases when determined by diastolic BP. 2) In 28 patients subjected to the intensive therapy, the control category of systolic BP was changed from 11 to 37% in well-controlled cases, from 42 to 30% in poorly-controlled cases, from 39 to 26% in masked hypertension cases and from 8 to 7% in white-coat hypertension cases. The shift in categories in both poorly-controlled and masked hypertension cases to the better category (well-controlled), was statistically significant (p=0.001, by Wilcoxon's signed rank test). Similarly, the control category of diastolic BP was changed from 45 to 43% in well-controlled cases, from 13 to 15% in poorly-controlled cases, from 34 to 32% in masked hypertension cases and from 8 to 10% in white-coat hypertension cases. There was a tendency for the prevalence of poorly-controlled and masked hypertension to improve to the well controlled category in response to intensive therapy (p=0.0625, by Wilcoxon's signed rank test). 3) The plasma concentration of plasminogen activator inhibitor (PAI-1)/tissue plasminogen activator (t-PA) complex (total PAI-1 complex) was significantly decreased after intensive therapy (17.3+/-7.8 ng/ml vs. 13.5+/-4.6 ng/ml, n=28, p<0.01 by paired t-test). In contrast, the plasma concentration of t PA was unchanged even after intensive therapy (4.8+/-3.9 ng/ml vs. 6.2+/-2.9 ng/ml, n=28, ns). These data suggest that home BP obtained in the morning is a useful measure for evaluating morning hypertension in PD patients, most of whom have refractory hypertension categorized as either poorly-controlled or masked hypertension. Intensive treatment with ACE-I/ARB given in the evening along with alpha1-blocker at bed time combined with a diuretic and/or long-acting CCB in the morning is efficacious in controlling the BP of patients with refractory hypertension in PD patients. The link between the reduction in plasma total PAI-1 levels and the intensive therapy may suggest that this therapeutic strategy could prevent thrombotic events associated with morning hypertension in patients on PD. PMID- 15754763 TI - [Survey of hemodialysis inpatients in long-term care facilities]. AB - OBJECTIVE: In Japan, the number of elderly patients on hemodialysis has markedly increased in recent years. We conducted a survey to confirm the current status of inpatient hemodialysis patients and dialysis facilities with the aim of providing better care. MATERIAL AND METHODS: The subjects were 57 dialysis patients admitted to the Jishu Hospital for three consecutive months or longer as of February 2004. The survey was conducted by obtaining informed-consent from the patients and their families. The patients were interviewed and a questionnaire was sent to their families. The survey items were as follows; 1) patient characteristics, 2) domestic status, 3) physical and mental condition and 4) awareness concerning the hospital. RESULTS: The survey showed that dementia is present in almost all elderly hemodialysis patients and they require some degree of assistance for the activities of daily living. In addition, 57.9% patients had already been admitted to or had visited two or more hospitals. The current status of inpatients had improved slightly. Although the period of hospitalization was longer than one year in 64.9% of respondents, it had decreased by around 13% in comparison with that in the previous survey at 1997. These results were considered to be caused by increases in inpatient dialysis facilities, aging related deaths, and by patients switching to home care after discharge because of the higher economic burden. In the interview, some patients were satisfied with inpatient life, but about one half of the patients said they wanted home care. Key caregivers for patients are mainly middle-aged or older family members (> 50 years old), indicating that caregivers are aging. On the other hand, the present study showed that hospitalization provides advantages for family life, such as improvements in peace of mind, life rhythm, leisure time and physical condition. CONCLUSION: Since the aging of dialysis patients results in a decrease in physical activity and progression of dementia, long-term hospitalization has a high potential to induce bedridden patients. In addition to the requirement for more long-term facilities, efforts should be made to improve physical activities and inhibit the progression of dementia. PMID- 15754764 TI - [Principles of surgical care organization and structural characteristics of sanitary casualties in counter-terrorist operations in the Northern Caucasus (Report I)]. AB - Two operational-and-tactical factors had the main influence on organization of the surgical care rendered to the casualties on the Northern Caucasus: the fulminant and maneuverable nature of combat operations at the beginning of both armed conflicts and rather small territory of war theatre. The main special features of organization of surgical care to the casualties were the use of Medical Squads of Special Purpose in the combat conditions and maximal approximation of the specialized surgical care to the wounded by echelonment of medical units and wide use of helicopter evacuation. The structure of sanitary losses was characterized by the increase in the frequency of mechanical traumas, thermal and combined injuries, by the high share of the casualties with head injuries. Besides the number of seriously wounded and critical casualties has increased. PMID- 15754765 TI - [Using modern informative technologies in the educational process]. PMID- 15754767 TI - [Autologous hemocomponents obtained in the multi-purpose military medical institution]. PMID- 15754766 TI - [The role of humoral markers of inflammation activity in the evaluation of adequacy of antibacterial chemotherapy for community-acquired pneumonia]. AB - To determine the significance of results of dynamic study of C-reactive protein (C-RP) and ceruloplasmin (CP) concentrations in plasma for evaluation of adequacy of community-acquired pneumonia (CAP) launching antibacterial chemotherapy we investigated 97 patients (servicemen on draft). C-RP and CP were determined by immunoturbidimetric method. The blood samples were obtained on admission, on the 3rd-4th day of treatment and on the 7th-8th day of stay in the hospital. The transformations of pulmonary infiltrates were evaluated at the same periods with the use of digital chest fluorography. The strong correlation between transformations of pulmonary infiltrates for serum C-RP level (r=0.79) and CP level (r=0.71) was observed. The dynamic assessment of C-RP and CP levels is a simple and reliable test that permits to use it as an adjunctive procedure in C RP diagnosis and evaluation of treatment effectiveness. The serum CRP level more than 110 mg/l and CP level higher than 6.3 mg/l should be considered as diagnostic and prognostic markers of severe CAP. PMID- 15754768 TI - [Madopar in the complex therapy of ischemic insult]. PMID- 15754769 TI - [The main directions of hormonal therapy of the service women]. PMID- 15754770 TI - [The pathogenetic substantiation of the method used for rapid detection of high predisposition to hypertensive disease in practically healthy young servicemen]. PMID- 15754771 TI - [Clinical and laboratory characteristics of HIV-infection in servicemen]. AB - HIV-infection in Russia and its Armed Forces is a serious threat to the national health and country's safety. Study of peculiarities of disease spread in the military collectives and HIV-infection manifestation is an actual problem. HIV infection is diagnosed during the latent stage in all categories of servicemen and characterized by unchanged function of cellular and humoral immunity. In most servicemen with HIV-infection the disease course has a form of co-infection with parenteral hepatitis B and C. PMID- 15754772 TI - [Medical and evacuation support of the Russian Army during the First World War]. PMID- 15754773 TI - [Professor A.A.Bobrov--the surgeon and clinician]. PMID- 15754774 TI - [The role and significance of phytotherapy during the Great Patriotic War]. PMID- 15754775 TI - An adipocentric view of liver fibrosis and cirrhosis. AB - Liver fibrosis is the consequence of chronic or repeated liver injury caused by hepatotoxic agents like alcohol and viruses, as well as immune and congenital metabolic disorders. Nonalcoholic fatty liver disease (NAFLD), caused by obesity and abnormal lipid metabolism, may be the latest known cause of liver fibrosis and cirrhosis. Furthermore, NAFLD with obesity can provide a terrain in which alcoholic and viral liver diseases, such as chronic hepatitis C, are prone to cause liver cirrhosis. Insulin, insulin-like growth factor (IGF)-1, peroxisome proliferator-activated receptors (PPARs), leptin, adiponectin, and preadipocyte factor-1/delta-like1 (Pref-1/dlk1) are hormones, growth factors, nuclear receptors, and cytokines that are actively involved in lipid metabolism. They share common target cells important in liver fibrosis, i.e., hepatic stellate cells (HSCs). Activation of HSCs is known to initiate and perpetuate liver fibrosis. Insulin and IGF-1 stimulate HSC activation and collagen production in vitro. However, IGF-1 alleviates liver fibrosis in vivo. Ligands of PPARy inhibit HSC activation and collagen synthesis in vivo and in vitro, and are helpful in decreasing liver fibrosis. But ligands of PPARbeta enhance proliferation of HSCs. Leptin is profibrogenic, and liver fibrosis is decreased in leptin- or leptin receptor-deficient mice. Adiponectin is, on the contrary, anti-fibrogenic. Extensive liver fibrosis may develop in adiponectin-knockout mice and is alleviated by administration of recombinant adiponectin. Pref-1/dlkl is implicated in fibrogenesis of the liver through its modulation of HSCs. The use of such biologically active molecules in lipid metabolism as ligands of PPARgamma and adiponectin might not help slim down a patient on the whole, but can potentially be used to halt the progression of liver fibrosis. Weight reduction, a strategy for controlling obesity and metabolic syndromes, may also be a tool for decreasing NAFLD and alleviating liver cirrhosis. PMID- 15754776 TI - The significance of mycobacterium tuberculosis antibody, antigen 60 IgG in patients with abnormal chest radiography. AB - BACKGROUND: The identification of acid-fast bacilli (AFB) in sputum or tissue is the definite diagnosis of tuberculosis. However, this method of diagnosis is restricted by certain limitations. The serologic diagnosis of tuberculosis has been used for a long time. The aim of this study was to determine the sensitivity and specificity of Antigen 60 (A60) immunoglobulin G (IgG) in patients with abnormal chest radiography and to assess its application in the serologic diagnosis of pulmonary tuberculosis. METHODS: Data on patients who had been diagnosed using results of culture and pathology as having active pulmonary tuberculosis (N=178), other non-tuberculosis pulmonary disease (N=34), or no pulmonary disease (N=117) was collected from January 2001 through December 2002. The data of A60 IgG using enzyme-linked immunosorbent assay (ELISA), chest radiography, tuberculosis culture and pathology were obtained retrospectively. The cutoff value of A60 IgG was chosen according to a receiver operating characteristic (ROC) analysis. The sensitivity, specificity, positive predictive value, negative predictive value and likelihood ratio for positive and negative test were determined. RESULTS: The chosen cutoff value of 261.2 units defined the sensitivity (49.4%) and specificity (79.5%) of the test. The positive predictive value and likelihood ratio were 95.7% and 4.20, respectively, for patients with abnormal chest radiography and 88.2% and 2.97, respectively, for patients with abnormal chest radiography and negative AFB in sputum smear. CONCLUSIONS: Because of the high positive predictive value and likelihood ratio, a positive A60 IgG test in the presence of an abnormal chest radiography can help make an accurate clinical diagnosis of pulmonary tuberculosis. PMID- 15754777 TI - Efficacy and adverse effects of patient-controlled epidural or intravenous analgesia after major surgery. AB - BACKGROUND: The purpose of this retrospective study was to determine whether epidural fentanyl-bupivacaine patient-controlled analgesia (PCA) was more efficacious and had fewer adverse effects than epidural or intravenous morphine PCA. METHODS: We retrospectively retrieved data from 859 patients (mean age 64+/ 7 years) who received continuous epidural medication, either morphine or fentanyl bupivacaine PCA, or intravenous morphine PCA for postoperative pain control after major elective surgery from 1999 to 2000. Pain was assessed postoperatively using a verbal analogue pain scale (VAS, 0-10) during rest, mobilization, and coughing. Adverse effects including nausea, vomiting, pruritus, urinary retention, sedation, motor block, and respiratory depression (< 8 breaths per minute) were recorded. On the third postoperative day, the overall quality of pain control was evaluated using a pain relief scale (PRS, 1-4). RESULTS: There were 201 patients who had epidural morphine PCA, 427 patients who had fentanyl-bupivacaine PCA, and 231 patients who had intravenous morphine PCA. Most patients (> 86%) who received epidural or intravenous PCA, either morphine or fentanyl combined with bupivacaine, experienced good pain relief (VAS, 0-3) during rest, mobilization, and coughing. Nonetheless, patients who received epidural morphine or fentanyl bupivacaine had greater satisfaction with overall pain relief (PRS = 4) than did those who received intravenous morphine (p<0.05). Nausea and vomiting were most common in the epidural morphine group (p<0.05). Pruritus occurred least often in patients who received epidural fentanyl-bupivacaine analgesia (p < 0.05). There were no differences in other adverse events such as urinary retention, sedation, and motor block among the three groups. No respiratory depression was found in any patient. CONCLUSIONS: Patients receiving epidural fentanyl-bupivacaine PCA experienced better overall pain relief, while morphine PCA, either epidurally or intravenously, caused more side effects. It is considered safe to use continuous epidural PCA with fentanyl-bupivacaine in patients receiving major elective surgery. PMID- 15754778 TI - Early predict the outcomes of refractive accommodative esotropia by initial presentations. AB - BACKGROUND: The differential diagnosis between fully and partially refractive accommodative esotropia (Ac-ET) depends on outcomes after intervention with refraction correction. Whether the differences exist in terms of initial clinical features between these two variants has not been fully explored. METHODS: Children between the ages of 6 months and 8 years with esotropia and spherical equivalent greater than +3.00 (D) were included in this study. After wearing diopters glasses for at least 2 years, children were classified according to the indexed criteria into the fully Ac-ET group (group A, N=28) partially Ac-ET (group or the B, N=17). Six clinical parameters, including age at onset, age at first visit, visual acuity, refractive error, angle of esodeviation, and presence or absence of inferior oblique overaction at initial presentation were compared between these two groups. RESULTS: The angle of esodeviation (31.4+/-11.6 PD vs. 42.6+/-12.6 PD, p = 0.004) was significantly different between the fully and partially Ac-ET groups, while refraction (+5.79+/-1.84 D vs. +4.79+/-1.40 D, p = 0.062) had borderline significance. On the contrary, the age at onset (2.35+/ 1.74 yrs vs. 2.01+/-1.96 yrs, p=0.539), age at first visit (3.51+/-1.36 yrs vs. 3.01+/-1.70 yrs, p =0.285), inferior oblique overaction (32% vs. 47%, p =0.317), and visual acuity before (LogMAR: 0.40+/-0.25 vs. LogMAR: 0.34+/-0.25, p =0.544) and after treatment (LogMAR: 0.057+/-0.079 vs. LogMAR: 0.051+/-0.19, p = 0.088) were similar in the two groups. CONCLUSIONS: Children with a smaller angle of esodeviation and higher hyperopia were more likely belonging to fully Ac-ET, which can be treated with glasses without the need of surgical intervention. Early detection and early treatment of accommodative esotropia are needed to prevent strabismus and amblyopia. PMID- 15754779 TI - Analysis of the health status of foreign brides in a community hospital in Taipei County. AB - BACKGROUND: Very few studies regarding the health status of foreign brides in Taiwan are so far available. The purpose of this study was to analyze the health status of foreign brides in a community hospital setting in Taipei County. METHODS: We retrieved and analyzed 493 foreign brides' medical records recorded between November 1, 2002 and October 31, 2003. The records included a medical history, physical examination parameters, serological test data for infectious diseases, urinalysis for drug use, stool sample data for intestinal parasites, and chest radiography mainly to rule out tuberculosis. RESULTS: Of the 493 foreign brides included in this study, 247 were from China (50.1%); 122 from Burma (24.7%), 55 from Indonesia (11.2%), and 32 from Vietnam (6.5%). A small proportion of women, 69 (14%), were infected with intestinal parasites. Seven subjects (1.4 %) had tuberculosis, of which 4 (0.8%) were diagnosed as having old tuberculosis. None of the foreign brides was positive for the HIV antibody or on the syphilis screening. The seronegative rate of anti-rubella IgG among the foreign brides was 14.4%. Three percent of the brides had an elevated morphine level (> or = 300 ng/mL), but none of them was positive (> or = 500 ng/mL) for amphetamine. CONCLUSIONS: Future national statistics should include brides from China. The main concerns about foreign brides are the low vaccination rate against rubella virus, the high infection rate with intestinal parasites, and the high prevalence of tuberculosis. A nationwide survey should be carried out in order to assess the health status of all foreign brides in Taiwan. PMID- 15754780 TI - Posterior decompression and stabilization for metastatic spine diseases. AB - BACKGROUND: The purpose of this study was to investigate the clinical results of posterior decompression and stabilization for metastatic diseases of the thoracolumbar spine. METHODS: From 1980 to 2001, 70 consecutive patients with spinal metastases underwent palliative surgery by posterior decompression of spinal cord and subsequent stabilization with instrumentation. There were 38 women and 32 men. Their ages ranged from 24 to 75 years (mean 58 years). We retrospectively reviewed medical records to analyze their survival, clinical presentations, image findings and surgical outcomes. RESULTS: Sixty-one patients (87%) survived longer than 3 months. Forty-nine patients (70%) survived longer than 6 months, of whom 35 patients were still alive at an average of 24 months (range 13-40 months) after surgery. All maintained spinal stability postoperatively. Forty-seven of 60 patients (78.3%) with severe pain obtained significant symptomatic relief for 3 months or more, and 38 of 54 (70.1%) paralyzed patients gained neural improvement. Of the 60 patients bedridden before surgery due to pain or paresis, 36 patients (60%) experienced an increase in activity tolerance. CONCLUSIONS: The results of this study shows that neurological recovery, pain relief and mobility can be enhanced by posterior decompression and stabilization in highly selective patients with spinal metastases. PMID- 15754781 TI - Esthetic restoration of infra-occluded retained primary mandibular incisors with all-ceramic crowns in adult dentition. AB - The prevalence of hypodontia is reported to be between 1.5% to 10% in the permanent dentition. In the anterior teeth, maxillary lateral incisors and mandibular central incisors are the most frequently involved teeth. This causes esthetic problems for the patient. Several reports have focused on restoration of retained maxillary primary anterior teeth, but none have described restoration of retained mandibular primary incisors. This clinical report describes the restoration of infra-occluded retained primary mandibular central incisors of a 17 year-old girl diagnosed with hypodontia. All-ceramic crowns made with computer aided design/ computer-aided manufacturing technology were used to restore the teeth incisally and interproximally. Due to a relatively short root length and inadequate crown-root ratio, the primary mandibular central incisors were splinted and adjusted to distribute the protrusive force evenly across the maxillary and mandibular incisors. Functional and esthetic results were achieved. PMID- 15754782 TI - A social skills training program for preschool children with attention deficit/hyperactivity disorder. AB - Many children with Attention-deficit/hyperactivity disorder (ADHD) have difficulties in regulating their emotions and maintaining good peer relations. It is important for children with ADHD to receive interventions as early as possible so that their social and emotional development can be achieved. Some social skills training programs for school-ages children with ADHD in Taiwan have shown positive results. However, such programs have not been applied to preschool children with ADHD yet and its applicability needs to be explored. In this report, we describe the clinical experience of conducting social skills training with 8 boys with ADHD (aged 4 to 6 years) and their parents. Eight weekly training sessions were conducted. Each session included a joint parent-child social skills training group (1 hour) and a parent group (40 minutes). After the training, most parents (75%) reported improvements in their children's behaviors. Clinical implications and limitations of this study are discussed. PMID- 15754783 TI - Paranasal sinus involvement in acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) is a hemopoietic malignancy of the bone marrow that rarely invades the sinonasal area. If infiltration of paranasal sinuses occurs, it may lead to rhinosinusitis and orbital complications that need aggressive treatment. In this report, a 26-year-old male patient who had a history of ALL and had one relapse, suffered from rapid progression of right periorbital pain and exophthalmos, which usually presented in patients with orbital complications of sinusitis. A sinus computed tomography showed right maxillary and ethmoid sinus opacification with orbital bone destruction. Urgent endoscopic sinus surgery (ESS) was performed for orbital decompression and histopathologic diagnosis. Pathology revealed lymphoblast infiltration and inflammation of respiratory mucosa. Subsequent bone marrow aspiration cytology confirmed the diagnosis of a second relapse of ALL. Obstructive causes of sinusitis should be evaluated in patients with ALL. ESS has proven to be effective in obtaining tissue for histopathologic diagnosis, and in treating patients with sinusitis with orbital complications. PMID- 15754784 TI - A solitary laryngeal neurofibroma ina pediatric patient. AB - A neurofibroma of the larynx is a rare disease that usually involves the arytenoids and the aryepiglottic fold. Pediatric patients with a laryngeal neurofibroma often present with progressive dyspnea. A definitive diagnosis of a laryngeal neurofibroma is based on the histopathologic demonstration of the characteristic spindle cells, and a positive result in immunohistochemical staining for S-100 protein. Tumor excision of laryngeal neurofibromas by an external approach was formerly common. We present a case of a solitary laryngeal neurofibroma in a 4-year-old child that was completely excised using direct laryngoscopy and a CO2 laser. There has been no recurrence after a follow-up of 4 years. For pediatric patients with an isolated laryngeal neurofibroma, endoscopic surgery provides an alternative approach due to its safety, effectiveness, and minimal invasiveness. PMID- 15754785 TI - [Guidelines for antimicrobial treatment and prophylaxis of urinary tract infections]. AB - Recommendations for antimicrobial treatment and prophylaxis of urinary tract infections (UTI) have been made according to the results of investigation of resistance of the most frequent causative agents of UTI to antimicrobial drugs. This investigation has been conducted for the past seven years by the Committee for monitoring bacterial resistance to antibiotics in the Republic of Croatia, with consensus of eight professional societies of the Croatian Medical Association. Uncomplicated cystitis is treated 1, 3, or 7 days, complicated 7 days, pyelonephritis 10-14 days, and complicated UTI 7 to 14 days, rarely longer. For the treatment of cystitis fluorokinolons, nitrofurantoin, betalactam antibiotics, and in the fields of lower resistance trimethoprim/sulfamethoxazol are being used. Single treatment with fluorokinolons is administered to otherwise healthy young women with normal urinary tract in whom cystitis symptoms have been present for less than 7 days. Empiric antimicrobial treatment of pyelonephritis, recurrent and all complicated UTI must be reviewed after urine culture finding is obtained. In the treatment of bacterial prostatitis and febrile UTI in males, the drug of first choice is ciprofloxacin. Asymptomatic bacteriuria (AB) is treated in pregnant women, newborns, preschool children with urinary tract abnormalities, before invasive urologic and gynecologic procedures, in kidney transplant recipients, and in the first days of short term urinary bladder catheterization. Recommendations for the treatment of AB in patients with diabetes mellitus have been controversial in the past two years. Antimicrobial prophylaxis is administered mostly one hour prior to the diagnostic or therapeutic invasive urological procedure, using selected antimicrobial agents. PMID- 15754786 TI - [Epidemiological study of psychiatric hospitalizations in Bjelovar General Hospital]. AB - Epidemiological studies in psychiatry contribute to the knowledge of prevalence of mental disorders and rational planning of protection of mental health, and also enable the application of optional methods in primary, secondary and tertiary prevention. Based on the information provided by the Register of psychiatric activities of the Bjelovar General Hospital an epidemiological study of the hospital treated patients for the years 1980, 1985, 1990, 1995, and 1999 was made. In determining the diagnostic groups the authors used the International classification of diseases ICD10 and references of the WHO, adapting all the diagnostic entities to the valid classification. According to the Register, in the observed years of the twenty-year period, 4631 psychiatric patients, who make the sample of the study, were treated. The fall of the general number of psychiatric hospitalizations by 28.8%, the reduction of the proportion of men and women in the sample, and also the reduction of mortality were noted. In the observed period the fall of the general number of hospitalised alcoholics, and the increase of hospitalisations due to dementia, schizophrenia, affective disorders and critical conditions were visible. The differences in the results between the initial and the final year of the epidemiological study were analysed by the chi square test. Changing of the social system, the war and post-war occurrences have led to the series of changes in the epidemiological image of psychiatric hospitalisations. The results of the epidemiological study will help the evaluation, rational planning and further development in the protection of mental health of the inhabitants of Bjelovar region. PMID- 15754787 TI - [Doppler ultrasonography of superficial lymph nodes]. AB - Normal lymph nodes (l.n.) are elongated and hypoechoic, with central echogenic hilum on B-mode, while color-doppler (CD) shows central hilar vascularization. Chronic inflammatory l.n. are elongated, with echogenic hilum, hilar vascularization on CD and resistance index (RI) 0.60-0.70. Acute lymphadenitis shows intensive hilar flow with RI<0.60; abscess of l.n. is anechoic with through transmission. Tuberculous l.n. are inhomogenously hypoechoic and rounded, with deformed hilar vascularization on CD, occasionally with centripetal flow. They may mimick malignant l.n. on CD due to vessel dislocation by granulomatous masses. L.n. in stage of micrometastasis are of similar structure as reactive l.n., with preserved hilar vascularization; as the metastasis grows, focal cortical expansion and/or destruction of intranodal structure with dislocation and compression of blood vessels may occur; vascular resistance is consecutively increased; in advanced stage of metastatic growth l.n. becomes rounded, hilum is not visible, cortex is heterogenous, occasionally with hyperechoic keratin deposits, unsharp nodal margin is the sign of extranodal neoplastic spread; mainly peripheral vascularization, with fragmentation, deformation and dislocation of intranodal vessels are seen on CD, with increased RI>0.80; necrotic areas are anechoic and avascular on CD. Lymphomatous nodes have preserved internal structure, sharp margins, hypoechoic cortex, frequently with through transmission; large arborized hilum may be displaced; abundant hilar flow is seen on CD, with normal or slightly increased vascular resistance. Although the sensitivity of B-mode and Doppler features of l.n. is mostly limited (false negative rate is significant), high specificity of some of them establishes sonography as a valuable complementary method in differential diagnosis of lymphadenopathy: the presence of hilar vascularization with RI<0.40 accurately indicates acute lymphadenitis, while predominantly peripheral flow pattern and RI>0.80 are typical of metastatic l.n. High enddiastolic velocities (EDV>9 cm/s) are only rarely seen in metastases, while very low EDV<1 cm/s indicates metastatic lymphadenopathy. As sonography is very suitable imaging method for puncture guidance, B-mode and CD analysis of l.n. need to be combined with aspiration cytology whenever possible, as it may considerably improve the accuracy of non-invasive patient diagnostic work-up. Literature on sonography of l.n. was reviewed, and selection of most relevant articles was made. PMID- 15754788 TI - [Bronchial impairment of main bronchus by aspiration of ferrous sulphate tablet]. AB - The late occurrence of broncial impairment in two children who had aspirated ferrous sulphate tablet ih presented. Local release of cytotoxic oxidant radicals led up to tissue necrosis and consequently to bronchial stenosis and obliteration. Bronchoscopy in the first child confirmed total obliteration of the left main bronchus eleven months after aspiration of iron tablet, and pulmectomy had to be done. In the second child five months after aspiration of iron tablet bronchoscopy confirmed severe stenosis of the right main bronchus and atelectasis of the right lung lobe. Reventilation of right lung has been achieved one month after administration of antibiotics, metilprednisolon and physical therapy. The patient underwent laser therapy (few times) and baloon dilatation in the University Clinic in Graz, but it did not hold passability of the right bronchus. Reventilation of the right middle and lower lobe (but not upper lobe) has been achieved by insertion of endobronchial stent in the main right bronchus. Formation of granulomatous tissue at stent borders necessitated further laser therapy and baloon dilatation. Treatment of this child must be continued. PMID- 15754789 TI - [Nonalcoholic steatohepatitis in children]. AB - Nonalcoholic steatohepatitis (NASH) occurs not only in adults but also in children. NASH is diagnosed in obese children with increased aminotranspherases, characteristic ultrasonography and typical percutneous liver biopsy (fat vacuoles in hepatocytes, different stages of inflammation with or without liver fibrosis) when other possible causes of steatosis are excluded. It is important to diagnose NASH and start the treatment as early as possible in order to prevent irreversible liver damage. The first line therapy consists of reduction of body weight by low-calorie diet and exercising. Ursodeoxycholic acid has been used as adjuvant therapy mainly in adults, and vitamin E also in children. We presented 5 cases with NASH (2 girls, 3 boys; age 11-15 years). All were obese and had increased transaminases, and ultrasonography and liver biopsy typical for NASH. With low-calorie diet in all our patients BMI declined, symptoms vanished and transaminases decreased (in two patients transaminases normalized). PMID- 15754790 TI - [Laparoscopic surgery during pregnancy]. AB - With the development oflaparoscopic surgery, number of contraindications has become less and less. At the beginning of laparoscopic surgery pregnancy was considered an absolute contraindication to laparoscopic surgery, because of unknown influence of CO2-pneumoperitoneum on fetus. Over the past few years, in literature there have been several case reports of successful laparoscopic surgery in pregnancy. Since 1992 when the first laparoscopic cholecystectomy was done at the University Department of Surgery of Sveti Duh General Hospital in Zagreb, three pregnant women were managed successfully laparoscopically. Two laparoscopic cholecystectomies, and one laparoscopic appendectomy during pregnancy, were performed. RESULTS: In all three patients, there were no complications during and after surgery. Pregnancies were completed at term spontaneously, delivering healthy children. PMID- 15754791 TI - [Cognitive impairment in multiple sclerosis patients]. AB - In addition to neurological symptoms, multiple sclerosis is characterized by cognitive function impairment. Disturbances of memory, recall, information processing, visual-spacial perception, attention, and executive function, in less extent of speech, are present in about 60% of patients. They are similar to disorders in other subcortical dementias. Once they appear, they rarely recede. Conventional, and especially nonconventional magnetic resonance imaging evaluates more precisely the tissue substrate--diffuse neuroaxonal lesion of the entire brain parenchyma--than clinical findings, already in the early stage of the disease. Alterations in the brain imaging are manifested by T2 hyperintensive and T1 hypointensive lesions, decreased neuronal marker N-acetyl-aspartate in magnetic spectroscopy, decreased magnetization transfer ratio, and increased diffusivity with reduced anisotropy in diffusion-weighted imaging. Total volume of brain lesion, corpus callosum diameter, and relation of measures of brain chambers and the rest of the brain, are best indices of cognitive dysfunction in multiple sclerosis. Their diagnosis in the very beginning of the disease allows early application of therapeutic procedures. Symptomatic treatment of these disorders is not efficient, and immunomodulation, particularly the use of biologic versions of interferon-beta, shows disputable effects. Cognitive dysfunctions affect relationships and working ability of patients. PMID- 15754793 TI - [Psychiatric epidemiology in Croatia]. AB - Psychiatric epidemiology is a relatively young medical discipline which brings up basic principles of classic epidemiology in research of mental disorders. Beginnings of psychiatric epidemiology in Croatia have its roots in the Royal Institute for Mentally Ill in Stenjevec (Kraljevski zemaljski zavod za umobolne u Stenjevcu) (today's Psychiatric Hospital Vrapce in Zagreb), which from its early days, pays special care to medical statistics e.g. psychiatric epidemiology. In anticipation of the 125th anniversary and 125 years of continuous work of this biggest psychiatric institution in Croatia, authors are presenting the review of this important field of psychiatry, through Croatian publications and professional-scientific work of Croatian authors. The majority of publications are dealing with epidemiology of endogenous psychoses e.g. schizophrenia, but epidemiology of alcoholism, neuroleptic malignant syndrome and (in recent time) posttraumatic-stress disorder are alfo present. PMID- 15754792 TI - [Pharmacotherapy of Alzheimer's disease--an evidence based approach]. AB - The aim of this article is to provide evidence-based recommendations for early diagnosis of Alzheimer's disease (AD) and hereby to give clinican guidelines for optimal detection of patients with AD. Our intention is also to unify diagnostic schemes in accordance with our objective and specific possibilities. Basic diagnostic procedures primarily are anamnesis and clinical examination with rational usage of neuroimaging, electrophysiological and laboratory procedures. Using these guidelines in medical practice in Croatia would be a good basis for future epidemiological and clinical multicentric studies. PMID- 15754794 TI - [Relations between Croatian and Slovenian Medical Societies from their beginings according to some published sources]. PMID- 15754795 TI - Adaptive spectral strain estimators for elastography. AB - In conventional elastography, internal tissue deformations, induced by external compression applied to the tissue surface, are estimated by cross-correlation analysis of echo signals obtained before and after compression. Conventionally, strains are estimated by computing the gradient of estimated displacement. However, gradient-based algorithms are highly susceptible to noise and decorrelation, which could limit their utility. We previously developed strain estimators based on a frequency-domain (spectral) formulation that were shown to be more robust but less precise compared to conventional strain estimators, In this paper, we introduce a novel spectral strain estimator that estimates local strain by maximizing the correlation between the spectra of pre- and postcompression echo signals using iterative frequency-scaling of the latter; we also discuss a variation of this algorithm that may be computationally more efficient but less precise. The adaptive spectral strain estimator combines the advantages of time- and frequency-domain methods and has outperformed conventional estimators in experiments and 2-D finite-element simulations. PMID- 15754796 TI - Spectral normalization for ultrasonic contrast microbubble detection. AB - Ultrasonic contrast agents consisting of microbubbles are used to assess tissue perfusion. The microbubbles are highly reflective and nonlinear and thus produce harmonics that are stronger than those from tissues. However, the magnitude of harmonic signals resulting from a region with microbubbles also depends on the acoustic pressure of incident ultrasound and the attenuation of intervening tissues in the ultrasound path. Therefore, the harmonic magnitude, as used in traditional harmonic imaging, may not be a reliable indicator of the presence or absence of microbubbles, and hence, tissue perfusion. To compensate for these effects, we present two parameters defined as the ratio of the harmonic to the fundamental component (HFR) and the ratio of the harmonic to squared fundamental (HSFR). A simplified model is used to illustrate the usefulness of these two parameters. Experiments show that both parameters improve detection of microbubbles and that HSFR performs better than HFR. PMID- 15754797 TI - Recent developments in tissue-type imaging (TTI) for planning and monitoring treatment of prostate cancer. AB - Because current methods of imaging prostate cancer are inadequate, biopsies cannot be effectively guided and treatment cannot be effectively planned and targeted. Therefore, our research is aimed at ultrasonically characterizing cancerous prostate tissue so that we can image it more effectively and thereby provide improved means of detecting, treating and monitoring prostate cancer. We base our characterization methods on spectrum analysis of radiofrequency (rf) echo signals combined with clinical variables such as prostate-specific antigen (PSA). Tissue typing using these parameters is performed by artificial neural networks. We employed and evaluated different approaches to data partitioning into training, validation, and test sets and different neural network configuration options. In this manner, we sought to determine what neural network configuration is optimal for these data and also to assess possible bias that might exist due to correlations among different data entries among the data for a given patient. The classification efficacy of each neural network configuration and data-partitioning method was measured using relative-operating-characteristic (ROC) methods. Neural network classification based on spectral parameters combined with clinical data generally produced ROC-curve areas of 0.80 compared to curve areas of 0.64 for conventional transrectal ultrasound imaging combined with clinical data. We then used the optimal neural network configuration to generate lookup tables that translate local spectral parameter values and global clinical-variable values into pixel values in tissue-type images (TTIs). TTIs continue to show cancerous regions successfully, and may prove to be particularly useful clinically in combination with other ultrasonic and nonultrasonic methods, e.g., magnetic-resonance spectroscopy. PMID- 15754799 TI - Real-time calibration of temperature estimates during radiofrequency ablation. AB - Radiofrequency ablation is an interstitial focal ablative therapy that can be used in a percutaneous fashion and permits in situ destruction of hepatic tumors. Recurrence rates after rf therapy are as high as 34-55%, due to difficulties in accurately identifying the zone of necrosis (thermal lesion) because of the low intrinsic acoustic contrast between normal and ablated liver tissue. Our goal is to provide real-time ultrasonic tracking of temperature changes over the large range of temperatures traditionally used (40-100 degrees C) in rfablation procedures using an external ultrasound transducer. Temperature estimates are obtained using a cross-correlation algorithm applied to rf ultrasound echo signal data acquired at discrete intervals during heating. Apparent tissue displacement estimates obtained at these discrete time-intervals are accumulated to obtain a cumulative displacement map, whose gradient provides after appropriate scaling provides a temperature map at the specified elapsed ablation duration. Temperature maps are used to display the initial temperature rise and to continuously update a thermal map of the treated region. In this paper, we develop calibration curves that relate the echo shift due to the change in the speed of sound and thermal expansion to the corresponding temperature increase on in-vitro tissue specimens. These calibration curves can then be utilized for the real time calibration and analysis of temperature estimates obtained from the rf echo signals during ablation. Temperature maps obtained using the calibration curve compare favorably to temperature estimates observed using the invasive thermosensor readings on the ablation electrode and previous results that utilized a linear calibration factor. PMID- 15754798 TI - Real-time 3D color flow Doppler for guidance of vibrating interventional devices. AB - The goal of this investigation was to examine the feasibility of guiding interventional devices using piezoelectric buzzers to create velocity sources, which were imaged and tracked with real-time 3D color flow Doppler. The interventional devices examined in this study included a pacemaker lead, Brockenbrough needle for cardiac septal puncture, cardiac guidewire and radiofrequency ablation needles for cancer therapy. Each was mechanically coupled to a piezoelectric buzzer and was imaged using a commercial real-time 3D ultrasound system with either a 2.5 MHz matrix array transducer or a 5 MHz, 22 F catheter transducer equipped with a tool port. In vitro images acquired in tissue phantoms, excised liver with a 'tumor' target and an excised sheep heart show strong vibration signals in 3D color flow Doppler, enabling real-time tracking and guidance of all the devices in three dimensions. In a sheep model, in vivo tracking of the pacing lead was performed in the superior vena cava as well as the right atrium using RT3D color flow Doppler images. The vibrating rf ablation needles were guided through the liver toward "tumor" targets in vivo with real time 3D color flow Doppler images. PMID- 15754800 TI - Comparison of fluorometric detection methods for quantitative polymerase chain reaction (PCR). AB - In this study, we compared the sensitivity of two different detection methods for quantitative polymerase chain reaction (PCR). Various amounts of a 75 mer single stranded deoxyribonucleic acid (DNA) fragment, which can be used as a DNA label for the immuno-PCR (iPCR) assays, were amplified by PCR. The amount of amplified DNA fragments was determined by the fluorescence (FL) of SYBR Green dye that specifically interacts with double-stranded DNA fragments. In the first selected detection method, real-time PCR, FL measurements were carried out at each thermal cycle, as the DNA was being amplified by PCR. This was achieved using the Applied Biosystems (ABI) Prism 7000 Sequence Detection System and its standard protocol. In the second detection method, referred to as end-point detection, after the PCR amplification was completed, off-line FL measurements were subsequently carried out using a conventional plate reader. In order to achieve the lowest limit of detection (LOD) from the off-line measurement, we have optimized a wide variety of parameters. Our data have indicated the LOD of real-time PCR method was approximately three orders of magnitude lower than the end-point measurement method, with a linear range spanning six orders of magnitude; 10 fmol to 10 zmol of PCR template. The lower LOD of the real-time PCR method could be partly due to the ability to maximize the number of thermal cycles that could be carried out in PCR, without increasing the non-specific amplification of any contaminating DNA. The results of this study can be applied to the development of ultra-sensitive iPCR assays for various disease markers. PMID- 15754801 TI - Development of a PAN-specific, affinity-purified anti-acetylated lysine antibody for detection, identification, isolation, and intracellular localization of acetylated protein. AB - Acetylation on the lysine residue is an important event of posttranslational modification of proteins. In this study, we developed a simple method to produce and to affinity purify the specific anti-acetylated lysine polyclonal antibody, which is useful for the detection, identification, isolation, and intracellular localization of acetylated proteins on the lysine residues. We utilized the chemically acetylated hemocyanin of keyhole limpets (KLH) as an immunogen to raise the immune serum and to isolate the population of the acetylated lysine specific antibody using the immobilized acetylated lysine as immunoaffinity ligand. The isolated antibody was tested to be useful for ELISA, immunoblotting detection, immunofluorescent localization, and affinity isolation of the acetylated proteins. PMID- 15754802 TI - Synthesis and analytical characteristics of new progesterone skeleton haptens. AB - This article describes the synthesis of new conjugated progesterone-bovine serum albumin substances, used to obtain specific antisera toward this hormone and for progesterone assay kits. In general, these kits are designed to be used on the farm or at the local veterinary clinics. PMID- 15754803 TI - Development of a simple method for rapid isolation of polymorphonuclear leukocytes from human blood. AB - Polymorphonuclear leukocytes (PMNs; commonly known as neutrophils) play essential roles in innate immunity and inflammation. Although there are standardized methods for the isolation of human neutrophils, they are time consuming and demand considerable technical expertise, making them unfeasible for many clinical applications. Here, we describe a simple and time-efficient technique for the isolation of human neutrophils, which adapts a readily available commercial cell preparation tube (CPT) currently in use for isolation of peripheral blood mononuclear cells (PBMC) and plasma and is now adapted to also yield neutrophils. The total time required for neutrophil isolation was less than 1 hr. Neutrophils isolated by this method were highly purified (> or =97%) as assessed by surface expression of the neutrophil specific marker, CD66b. Neutrophils isolated by this method were functional as demonstrated by their ability to secrete interleukin-1 receptor antagonist (IL-1RA). Neutrophils isolated using this new technique secreted significant amounts of soluble IL-1RA (929.3+/-197 pg/10(6)cells/mL) in response to lipopolysaccharide (LPS). Use of this adapted CPT method allows simultaneous isolation of functional human neutrophils as well as PBMC and plasma. Adoption of this new method will allow the conduct of different neutrophil assays at any clinical site without requiring trained laboratory personnel or a large staff time commitment. PMID- 15754804 TI - Development of an ELISA test for determination of the urinary trypsin inhibitor: analytical performance and applications. AB - Increased urinary excretion of urinary trypsin inhibitor (UTI) has been reported in various inflammatory conditions and in Alzheimer's subjects, but its diagnostic potential remains to be elucidated. A reliable and specific enzyme linked immunosorbent assay (ELISA) test for the determination of the UTI in human urine was developed. This assay was performed using 96-well microtiter plates. The plate surface is coated with an anti-UTI polyclonal antibody, the urine sample was added in a dilution range, and the detection was achieved using the enzyme-conjugated antibody. The assay was quantified by the build-up of colored product upon the addition of the substrate. Recoveries were 93%, and the intra- and inter-assay CVs were 4.25% and 21%, respectively. The ELISA showed parallelism of standard and urine samples and no significant interference by the biological matrix. The usefulness of the assay has been demonstrated by applying it to urine samples from Alzheimer's disease patients, and comparing with negative controls. UTI urinary levels are significantly increased in Alzheimer's subjects. PMID- 15754805 TI - Extraction, purification, and development of an enzyme-linked immunosorbent assay for osteocalcin. AB - The present study describes the isolation and purification of osteocalcin (OC) from bovine bones and the development of an enzyme-linked immunosorbent assay (ELISA) for OC as a marker of bone formation, for assessing bone health. Bone proteins were extracted from about 90 g of bovine bone powder using 20% formic acid. The protein extract was fractionated by gel permeation chromatography on Sephadex G-50 column followed by fast protein liquid chromatography (FPLC) on a MONO-Q column. The immunoreactive active fraction was then purified by chromatofocusing, using FPLC on a MONO P column and a single homogeneous band of molecular size of about 5.8kDa, as judged by Tricine SDS-PAGE following silver staining of the gel, was obtained. It reacted specifically with its antibodies in an ELISA. About 678 microg of purified OC was yielded from about 90 g of bovine bones. The purified OC was subsequently used for the raising antisera, which was used in the development of an indirect ELISA. The developed ELISA has a sensitivity of 2.5-4.0 ng/mL and was used in estimating levels of OC in women of various age groups. PMID- 15754806 TI - Development of coated tubes RIA for serum T3 (tri-iodothyronine) for production scale. AB - A coating procedure that could provide immobilization of antibodies, with increased binding capacity, that is cost effective, simple, robust, and appropriate for production scale application, is described. This coating approach of T3 antibodies to the polystyrene tubes has been systematically investigated to determine its utility for the development of coated tube Radioimmunoassay (RIA) for T3 in human serum. Further, the results obtained by the developed coating procedure are found to be comparable with those obtained by the "gold standard," the liquid phase RIA for T3. The coating procedure is completed in three major steps, each step involving an overnight incubation. The normal rabbit gamma globulins are physically adsorbed onto the polystyrene tubes and incubated. After washing, a second antibody (goat anti-rabbit antiserum) is added and incubated. To this antigen specific antibody is added (T3 antibody produced in rabbit) and further incubated. Finally, the non-specific sites on the tubes are saturated by the blocking solution. The concentration of normal rabbit globulin, titers of second antibody and T3 antibody, and time required for coating are optimized to arrive at a suitable coating protocol. The coated tubes were evaluated for precision, reproducibility, and stability. Various parameters such as total reaction volume, incubation time and temperature, total number and volume of washings, concentration of 8-anilino-1-naphthalene sulfonic acid (ANS), and quantity of tracer per tube are optimized to arrive at a suitable standard curve. The optimized assay is validated for the quality control parameters such as intra and inter-assay variations, recovery, and parallelism. The developed coated tubes assay had an assay range of 0.3-4.8 ng/mL with a sensitivity of 0.3 ng/mL at 90% B/B0. Batch to batch variation in coating was < 10%. The coated tubes were stable up to 1 year, which is adequate for production scale. PMID- 15754807 TI - 'Disruptive innovation' could change the health care landscape. PMID- 15754808 TI - EMR vs. DIMS. Yea: full EMR offers enhanced efficiency and workflow. PMID- 15754809 TI - Staff for success. PMID- 15754810 TI - Save yourself! The importance of disaster recovery. PMID- 15754811 TI - Not just any road. Get new physicians headed in the right direction. PMID- 15754812 TI - Thoughtful change. A service excellence focus for a billing system conversion. PMID- 15754813 TI - Recognizing the primary care 'nose dive'. PMID- 15754814 TI - Quality quagmire. Why systems and processes are important to your practice but often ignored. AB - The overuse, underuse and misuse of medical procedures in our nation is staggering. The Institute of Medicine, in two seminal reports, has brought to public attention the crisis in quality that afflicts the American medical system. Until recently, the movement toward quality has focused on hospitals, but now medical practices are joining in. The Centers for Medicare & Medicaid Services, the Medical Group Management Association Center for Research, individual medical groups and others are pushing for studies of health care quality in the ambulatory setting and for solutions that keep patients safer. PMID- 15754815 TI - Playing nice. Noncompete terms in physician employment agreements. AB - Medical groups often include a noncompete clause in their employment contracts with physicians to protect themselves if physicians leave and seek to open competing medical offices. Noncompete agreements can perform a valuable function by allowing medical groups to employ, train and financially support new professionals without fear that they are creating competitors, but the agreements' enforceability depends on state law. PMID- 15754816 TI - One-stop shopping pays off for high-tech orders. PMID- 15754817 TI - Hospitals keep watchful eye on sterilizer. PMID- 15754819 TI - Hadamard transform CE-UV detection for biological samples. AB - A Hadamard transform-capillary electrophoresis-UV (HT-CE-UV) detection technique is described for the analysis of biological samples. Pseudorandom injections of sample and buffer according to a simplex matrix obtained from the corresponding Hadamard matrix is performed with conventional capillaries. Alternating injections are achieved with a novel capillary "T" connector created by drilling conventional capillary dimensions through a 1-cm diameter polycarbonate disc. This connector design coupled with a switching system allows for rapid, electrokinetic injections of solution into alternating sample and buffer capillary arms for UV detection. The standard mixtures of nitric oxide (NO) metabolites, nitrite and nitrate, dissolved in physiological saline solution are injected into the separation capillary according to an 83-element injection sequence to obtain a signal-to-noise ratio (S/N) enhancement of ca. 4.5 over a single injection. Nitrite, being the less concentrated metabolite in NO detection and thereby more difficult to detect, was calibrated with the HT-CE-UV method and a limit of detection (LOD) of 0.56 microM was obtained. Rat blood plasma was analyzed with this detection system and demonstrated to be comparable with NO metabolite concentrations of previously published results. This HT-CE-UV method is described where a unique reservoir tube design that contains 8-microL standard nitrite sample volumes is placed over the end of the capillary arm to explore low volume limits for biological samples. PMID- 15754818 TI - Determination of binding constants and stoichiometries for platinum anticancer drugs and serum transport proteins by capillary electrophoresis using the Hummel Dreyer method. AB - A CE method has been developed to evidence and quantitatively characterize the interaction between platinum-based antitumor drugs and human serum proteins. This method is a variant of affinity CE modified regarding both experimental setup and data treatment so as to measure the peaks (or vacancies) that correspond to the bound drug when it slowly binds to the protein. Using the formalism of the Hummel Dreyer method and cisplatin and oxaliplatin as test compounds, a protocol for determining albumin and transferrin binding constants and stoichiometries, including (and distinguished by) 48 hours of incubation of the reaction mixture, was elaborated. Relative affinities of drugs toward different proteins in aqueous solution at physiological pH, chloride concentration, and temperature were compared in terms of overall binding constants and numbers of drug molecules attached to the protein. The results indicate that both platinum drugs bind to albumin more strongly than to transferrin, supporting the concept that the albumin fraction is a major drug supply route for chemotherapeutical needs. From a comparison with the binding parameters measured previously for cisplatin by other methods, conclusions were drawn about the validity of CE as a simple and convenient method for assaying protein-drug reactions with slow kinetics. PMID- 15754820 TI - Application of a capillary electrophoresis method for simultaneous determination of preservatives in pharmaceutical formulations. AB - Preservatives are used to protect pharmaceutical formulations from microbial attack during the period of administration to the patient. Because of their biological activity, preservatives have to be identified and assayed according to the same rules as apply to active components. A number of methods for separation of preservatives are reported, to account for the heterogeneity of their chemical structures. A capillary electrophoretic method was devised for simple and simultaneous qualification and quantification of the preservatives most often included in pharmaceuticals, such as benzyl alcohol, parabens, phenol, m-cresol, chlorobutanol, thimerosal. After systematic method development, the electrophoretic conditions were defined as: 50 mM borate buffer pH 9.0 containing 20 mM SDS. Separations were performed at a temperature of 20 degrees C and with detection at 214 nm. Preservatives under examination can be analyzed within a 10 min run. The method was successfully validated and applied to the determination of preservatives in a number of pharmaceuticals. Results from the CE method were compared with those from reference methods. PMID- 15754821 TI - Micellar electrokinetic chromatography for simultaneous determination of six corticosteroids in commercial pharmaceuticals. AB - We have developed a micellar electrokinetic chromatography (MEKC) method using bile salts for the simultaneous determination of six corticosteroids, including betamethasone, cortisone, prednisolone, 6alpha-methylprednisolone, triamcinolone, and prednisone. The separation was performed using borate buffer containing sodium cholate and sodium deoxycholate. Several parameters were studied, including bile salt concentrations, concentrations and pH of borate buffer, and analytical voltages. In method validation, calibration curves were linear over a range of 10-100 microM for each corticosteroid. The RSD (relative standard deviation) and RE (relative error) were all less than 5% for intra- and interday assays. The limit of detection of each analyte was 5 microM. The recoveries were greater than 95%. Application of this method for quality control of commercial tablets also proved to be feasible. All analytical values fall within the labeled amount of 90-110% for betamethasone and prednisolone, and of the labeled amount of 92.5-107.5% for 6alpha-methylprednisolone, as required by the United State Pharmacopeia 25 (USP 25). PMID- 15754822 TI - Analysis of selected ionic liquid cations by ion exchange chromatography and reversed-phase high performance liquid chromatography. AB - The chromatographic behavior of 8 ionic liquids - 7 homologues of 1-alkyl-3 methylimidazolium and 4-methyl-N-butylpyridinium - has been investigated with a strong cation exchange adsorbent. In particular, the dependence of the retention properties of these solutes on mobile phase composition, pH, and buffer concentration was evaluated with the aim of optimizing and improving the selectivity and retention of solute separation. While using the SCX stationary phase, several interactions occurred with varying strengths, depending on the mobile phase composition. Cation exchange, nonspecific hydrophobic interactions, and adsorption chromatography behavior were observed. Reversed phase chromatography occurred at low concentrations of acetonitrile, electrostatic and adsorption interactions at higher organic modifier concentrations. Elevated buffer concentrations lowered the retention factors without affecting the selectivity of ionic liquids. Obtained results were further compared to the chromatographic behaviour of ionic liquids in the reversed phase system. All analyzed ionic liquids follow reversed-phase behavior while being separated. Much lower selectivity in the range of highly hydrophilic compounds is obtained. This suggests preferred use of ion chromatography for separation and analysis of compounds below 4 carbon atoms in the alkyl side chain. PMID- 15754823 TI - Novel immobilized zinc(II) affinity chromatography for phosphopeptides and phosphorylated proteins. AB - Immobilized metal ion affinity chromatography (IMAC) is now a widely accepted technique for the separation of natural or artificial products that is beginning to find industrial applications. Here, we introduce a novel procedure for the separation of phosphopeptides and phosphorylated proteins by immobilized zinc(II) affinity chromatography. The phosphate-binding site of the affinity gel is an alkoxide-bridged dinuclear zinc(II) complex, the 1,3-bis[bis(pyridin-2 ylmethyl)amino]propan-2-olato dizinc(II) complex (Phos-tag), which is linked to a highly cross-linked 4% (w/v) agarose. The affinity gel (Phos-tag agarose) was prepared by the quantitative reaction of N-hydroxysuccinimide-activated Sepharose and a Phos-tag derivative having a 2-aminoethylcarbamoyl group in dry CH3CN. Phosphopeptides were retrieved in a quantitative and highly selective manner by a spin column method using Phos-tag agarose at room temperature. Furthermore, in this study, we demonstrate a simple, rapid, and reusable affinity column chromatography for the separation of phosphorylated proteins such as ovalbumin, alpha(s1)-casein, and beta-casein at physiological pH. PMID- 15754824 TI - Enantiomeric separation of chiral polychlorinated biphenyls on beta-cyclodextrin capillary columns by means of heart-cut multidimensional gas chromatography and comprehensive two-dimensional gas chromatography. Application to food samples. AB - Three commercially available chiral capillary columns, Chirasil-Dex, BGB-176SE, and BGB-172, have been evaluated for the separation into enantiomers of the 19 chiral polychlorinated biphenyls (PCB) congeners stable at room temperature. The enantiomers of 15 chiral PCBs were, at least to some extent, separated using these beta-cyclodextrin based columns. Multidimensional techniques, such as heart cut multi-dimensional gas chromatography (heart-cut MDGC) and comprehensive two dimensional gas chromatography (GC x GC), were investigated for their ability to solve coelution problems with other PCBs present in commercial mixtures and real life samples. Heart-cut MDGC improved the separation as compared to one dimensional GC, and enantiomeric fractions of the investigated chiral PCBs could be determined free from interferences. However, limitations on the number of target compounds that can be transferred to the second column in a single run and, therefore, the time consumption, have led to the evaluation of GC x GC as an alternative for this type of analysis. With GC x GC, two column set-ups were tested, both having a chiral column as first-dimension column, and two different polar stationary phase columns in the second dimension. On using both column combinations, congeners 84, 91, 95, 132, 135, 136, 149, 174, and 176 could be determined free from coelutions with other PCBs. Results on the application of heart-cut MDGC to food samples such as milk and cheese are given, as well as the first results on the application of GC x GC to this type of samples. PMID- 15754825 TI - Rapid determination of C6-aldehydes in tomato plant emission by gas chromatography-mass spectrometry and solid-phase microextraction with on-fiber derivatization. AB - A simple, rapid, sensitive, and solvent-free method was developed for determination of plant-signalling compounds, the three C6-aldehydes hexanal, (Z) 3-hexenal, and (E)-2-hexenal, in tomato plant emission by gas chromatography-mass spectrometry (GC-MS) and solid-phase microextraction (SPME) with on-fiber derivatization. In this method, O-2,3,4,5,6-(pentafluorobenzyl)hydroxylamine (PFBHA) in aqueous solution was first headspace adsorbed onto a 65 microm poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) fiber at 25 degrees C for 5 min, and then the fiber with adsorbed PFBHA was used for headspace extraction of tomato plant emission at 25 degrees C for 6 min. Finally, the resulting oximes adsorbed on the fiber were desorbed and analyzed by GC-MS. Extraction conditions and method validation were studied. The proposed method had low detection limit values for the three aldehydes from 0.1 to 0.5 ng/L and good precision (RSD less than 10%). In this work, the method was applied to investigation of tomato plant defense response to Helicoverpa armigera. PMID- 15754826 TI - Solid phase microextraction ion mobility spectrometer interface for explosive and taggant detection. AB - Ion mobility spectrometry (IMS) is a rugged, inexpensive, sensitive, field portable technique for the detection of organic compounds. It is widely employed in ports of entry and by the military as a particle detector for explosives and drugs of abuse. Solid phase microextraction (SPME) is an effective extraction technique that has been successfully employed in the field for the pre concentration of a variety of compounds. Many organic high explosives do not have a high enough vapor pressure for effective vapor sampling. However, these explosives and their commercial explosive mixtures have characteristic volatile components detectable in their headspace. In addition, taggants are added to explosives to aid in detection through headspace sampling. SPME can easily extract these compounds from the headspace for IMS vapor detection. An interface that couples SPME to IMS was constructed and evaluated for the detection of the following detection taggants: 2-nitrotoluene (2-NT), 4-nitrotoluene (4-NT), and 2,3-dimethyl-2,3-dinitrobutane (DMNB). The interface was also evaluated for the following common explosives: smokeless powder (nitrocellulose, NC), 2,4 dinitrotoluene (2,4-DNT), 2,6-dinitrotoluene (2,6-DNT), 2,4,6-trinitrotoluene (2,4,6-TNT), hexahydro-1,3,5-trinitro-s-triazine (RDX), and pentaerythritol tetranitrate (PETN). This is the first peer reviewed report of a SPME-IMS system that is shown to extract volatile constituent chemicals and detection taggants in explosives from a headspace for subsequent detection in a simple, rapid, sensitive, and inexpensive manner. PMID- 15754827 TI - Ultrasonic extraction followed by a novel filtration and clean-up device for screening of some polyphenols in tobaccos. AB - A simple and efficient HPLC method using a diode array detector (DAD) was developed for simultaneous determination of six polyphenols in tobaccos. Ultrasonic extraction was employed at 25 degrees C to extract the polyphenols present in tobaccos into anhydrous methanol. A novel filtration device linked to a clean-up cartridge was designed for simultaneous extract filtration and clean up. Optimized HPLC-DAD analysis, with multi-wavelength detection, was used for determination of the polyphenols. Because the content of some of the polyphenols is too low to be quantified directly, a concentration step was necessary. Anhydrous methanol was employed for extraction of the polyphenols because of its high extraction efficiency and its low boiling point in the concentration step. Using the proposed method, six polyphenols were quantified in tobaccos (Nicotina tobaccum L.). PMID- 15754828 TI - Selectivity of amylose tris(3,5-dimethylphenyl-carbamate) chiral stationary phase as a function of its structure altered by changing concentration of ethanol or 2 propanol mobile-phase modifier. AB - In a previous publication, solid-state NMR data showed that the structure of Chiralpak AD chiral stationary phase (CSP) was altered by changing the concentration of ethanol or 2-propanol modifier in the chromatographic mobile phase. This present paper reports the effect of the CSP structural change on chiral selectivity alpha. The enantiomers of a series of compounds were chromatographed using ethanol or 2-propanol in various concentrations as mobile phase modifier and the alpha values were determined. Changes of alpha were observed for some enantiomeric pairs when ethanol and 2-propanol concentrations were varied. These data correlate with previous findings on the structural changes of the CSP. Not every enantiomeric pair showed changes in alpha as the alcohol concentration was varied, indicating that the chiral selectivity depends not only on the CSP's structure, but also on the structures of the analytes. PMID- 15754829 TI - High-performance liquid chromatographic resolution of 1-(1,4-benzodioxane-2 formyl)-piperazine enantiomers after chiral derivatization. AB - Chiral separation of racemic mixtures is of the greatest importance to the pharmaceutical industry, as the isomers of a given racemate may exhibit substantially different pharmacological effects, not to mention possibly differing toxicity behaviour. A novel chiral separation method is developed for the determination of 1-(1,4-benzodioxane-2-formyl)piperazine (BFP) enantiomers. The indirect resolution is performed by applying precolumn derivatization with the chiral reagent 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC). The resulting diastereoisomers are separated on a reversed-phase ODS column with methanol-potassium dihydrogen phosphate (0.02mol/L, 50:50) as mobile phase. UV detection is at 250 nm. The effect of mobile phase composition upon resolution and analysis time is investigated. Two diastereoisomers show nearly base-line separation under optimal chromatographic conditions. The presented study provides a simple and accurate method for the enantiomeric quality control and the optical purity assay of BFP. PMID- 15754830 TI - Analysis of flunarizine in the presence of some of its degradation products using micellar liquid chromatography (MLC) or microemulsion liquid chromatography (MELC)--application to dosage forms. AB - The separation of flunarizine hydrochloride (FLZ) and five of its degradation products--1-[bis(4-fluorophenyl)methyl]-4-(3-phenyl-2-propenyl)piperazine, 4 oxide (A), bis(4-fluorophenyl)methanone (B), bis(4-fluorophenyl)methanol (C), 1 (3-phenyl-2-propenyl)piperazine(D), and 1-[bis-4-fluorophenyl) methyl] piperazine (E)--could be accomplished by reversed phase liquid chromatography using either micellar or microemulsion mobile phases. Cyanopropyl-bonded stationary phase has been used with UV detection at 254 nm. Microemulsion mobile phase consisting of 0.15 M SDS, 10% n-propanol, 1% n-octanol, and 0.3% triethylamine in 0.02 M phosphoric acid of pH 7.0, has been used for the separation of FLZ and its degradation products (B, C, D, and E). Micellar mobile phases consisting of 0.15 M sodium dodecyl sulphate (SDS), 10% n-propanol, 0.3% triethylamine (TEA) in 0.02 M phosphoric acid of pH values either 4.0 or 6.8 have been used for the separation of FLZ from its degradation products, i.e. either from (B, C, D, and E) or from (A, B, C, and D), respectively. Micellar liquid chromatography (MLC) was applied to the determination of FLZ in pure form as well as in dosage forms; the calibration graph was linear over the concentration range of 0.15-50 microg/mL with detection limit of 0.02 microg/mL (4.19 x 10(-8)M). PMID- 15754831 TI - A chronomic tree of life: ontogenetic and phylogenetic 'memories' of primordial cycles--keys to ethics. AB - A scientific optimization may become possible in ethics to the extent to which any reproducible since cyclic features of spirituality and of criminality become measurable. Should either or both the 'good' or the 'bad' be found to be at least passively influenced by cyclic physical environmental factors, as is putatively the case, these aspects of behavior may eventually become actively manipulable, perhaps utilizable for human survival. Toward this goal, chronomics has already mapped time structures in religious behavior that can lead to a study of underlying geographic/geomagnetic latitude-associated mechanisms. This paper, with further but clearly insufficient data, revealing the hurdle of relative brevity of the available time series constitutes a plea for much longer and denser worldwide time series, for further endeavors in various methods of analyses, some of which are promisingly available. PMID- 15754832 TI - About 21-year hale cyclic signature in agriculture complement those in biomedicine. AB - Data on total cultivated area and total production of wheat and rye, barley and two-row barley, corn, soya and sunflower in Romania (1968-2000) are re-analyzed from the viewpoint of chronomics. An about 10.7-year component is detected with borderline statistical significance. A 3-component model consisting of cosine curves, with periods of 3.62, 2.48 and 2.15 years, accounts for 27% of the overall variability. Transdisciplinary chronomics is advocated to systematically map predictable dynamics in agriculture such as Hale-like cycles in corn and soja and their interactions with chronomes of external environmental factors. PMID- 15754833 TI - Magnetic storm effect on the circulation of rabbits. AB - The ultrastructure of cardiomyocytes of rabbits was found to be drastically altered at a time coinciding with strong magnetic storms by comparison with that usually observed during quiet geomagnetic conditions. The circadian characteristics of systolic and mean pressure in the left and right ventricles of Chinchilla rabbits were assessed and compared between quiet and stormy magnetic conditions. Experiments repeated during four consecutive seasons at the times of equinoxes and solstices in the absence of magnetic storms were also examined for any circannual and/or transannual variation. The results have been interpreted in the broader context of non-photic influences on the circulation, fully supporting the presence of non-photic effects. The recording of magnetic activity in the laboratory, and until this is possible, the consultation of the physicists' routine recording of geomagnetic indices should become a sine qua non, since, as shown herein, magnetic storms can override the effect of the usually dominant synchronizer, the alternation of light and darkness. PMID- 15754834 TI - Does exposure to an artificial ULF magnetic field affect blood pressure, heart rate variability and mood? AB - The aim of this study was to determine whether an artificial magnetic field with an amplitude and frequency equivalent to those of geomagnetic pulsations during geomagnetic storms could affect physiology and psychology. Three healthy volunteers wore anambulatory BP monitor and an ECG recorder around the clock for 12 consecutive weekends in Winnipeg, Manitoba, Canada. In a room shielded against ELF and VLF waves, they were exposed for 8 hours per week to either a 50 nT 0.0016 Hz or a sham magnetic field at one of six circadian stages. Real exposure randomly alternated with sham exposure. They provided saliva and recorded mood and reaction time every 4 hours while awake. Systolic (S) and diastolic (D) blood pressure (BP), and heart rate (HR) were recorded every 30 minutes. Spectral analysis of HR variability (HRV) was performed using the maximum entropy method and a complex demodulation method. For these variables, daily means were compared between real and sham exposure, using paired t-tests. Their circadian MESOR, amplitude, and acrophase were analyzed and summarized using single cosinor and population-mean cosinor. Circadian rhythms were demonstrated for HR, SBP, DBP for sham exposure, salivary flow rate, positive affect, vigor, and subjective alertness (p < 0.001, -0.02). One participant showed higher HR, lower LF, HF, and VLF powers, and a steeper power-law slope (p < 0.005, -0.0001) in an early night exposure to the real magnetic field, but not in other circadian stages. There was no significant difference between circadian responses to real and sham exposure in any variable at any circadian stage. PMID- 15754835 TI - A real time non-invasive monitoring system for detection of hypovolemic state using RR interval variability in very low frequency ranges. AB - In offline analysis, very low frequency variability in RR interval and systolic arterial pressure has been reported below 0.1 Hz during head-up tilt (HUT) tests in conditions similar to hypovolemic states in humans. We designed a real time and non-invasive monitoring system of very low frequency RR interval variability to detect the hypovolemic state resulting from internal and external hemorrhages. Eight male Sprague-Dawley rats were subjected to monitoring before (normal state) and after withdrawal of 2 ml blood/100 g body weight over 15 minutes (hypovolemic state). Using one-channel electrocardiogram (ECG), the detection system monitored in real time the very low frequency components of RR interval variability using Fast Fourier Transform (FFT). Temperature mappings of rat abdomen were conducted simultaneously to monitor the hypothermic state after blood withdrawal using a local temperature mapping system with a deep body thermometer. This system demonstrated oscillation of the RR interval at 0.075 +/- 0.015 Hz in real time after blood withdrawal. The deep body temperature decreased significantly from 37.4 +/- 0.9 degrees centigrade to 35.5 +/- 1.2 degrees centigrade (p < 0.05) within 1 hour after blood withdrawal. Our monitoring system appears promising for the detection of hypovolemic state resulting from massive hemorrhage using a one channel ECG monitor. PMID- 15754836 TI - Periodic structures and diurnal variation in blood pressure and heart rate in relation to microgravity on space station MIR. AB - Four Russian crew members were studied on space station MIR, and blood pressure (BP) and heart rate (HR) data were continuously collected. BP and HR data were collected on earth 1 day before orbital flight to the space station, then at weeks 8, 16 and 24 during space flight, and again 1 or 2 days after returning to earth. Time serial data for BP and HR were analyzed by spectral analysis with the MemCalc system (Suwa Trust, Sapporo, Japan). Periodic structures of diurnal variation in systolic blood pressure (SBP), diastolic blood pressure (DBP) and HR were compared at 24-hour, 12-hour and 8-hour intervals, these being determined as the main periodic components for the assessment of BP and HR variability. The 24 h mean levels of SBP and HR during space flight were unchanged. Waking SBP was not different from pre-flight values. During sleep, in-flight changes in HR did not differ from pre-flight values. SBP during sleep in orbit increased to over pre-flight values. Waking DBP was reduced during flight. The SBP and HR phases over a 24-hour cycle were shortened with a more pronounced shortening in weeks 8 and 16 compared with pre-flight values, and at week 24 recovered to preflight values. The 12, 8-hour-cycle remained unchanged, and were similar to pre-flight values. At the space station, the astronauts' mission was carried out under strict control of sleeping and waking hours; therefore, their 24-hour schedule is an artificially constructed situation. Main periodicity structures were maintained by strict control of lifestyle during long-term space flight. The conclusions reached were as follows: 1) SBP levels during sleep in a space environment increased compared with those on earth; 2) the periodicity phase of BP and HR shifted toward to 24-hour cycle as a result of long-term space flight, even though these periods shortened after a few months compared with pre-space flight values. PMID- 15754837 TI - Effects of chronic sleep deprivation on autonomic activity by examining heart rate variability, plasma catecholamine, and intracellular magnesium levels. AB - Chronic sleep deprivation is associated with cardiovascular events. In addition, autonomic activity determined from the levels of the heart rate variability (HRV), plasma catecholamine, and intracellular magnesium (Mg) are important in the pathophysiology of cardiovascular events. This study therefore aimed to determine the effects of chronic sleep deprivation on autonomic activity by examining the HRV, plasma catecholamine, and intracellular magnesium levels. Thirty (30) healthy male college students ranging in age from 20 to 24 years of age (average 22 +/- 1 years; mean +/- SD) with no coronary risk factors such as hypertension, diabetes mellitus, hyperlipidemia or a family history of premature coronary artery disease (CAD) were included in the study. Over a 4-week period, the volunteers' plasma levels of epinephrine, norepinephrine, and erythrocyte-Mg were measured. The study was made during the 4 weeks before and immediately after college finals exams. HRV, obtained from 24-hour ambulatory ECG monitoring, included time and frequency domain indices. The HRV indices and erythrocyte-Mg decreased while norepinephrine increased during chronic sleep deprivation. It is concluded that chronic sleep deprivation causes an autonomic imbalance and decreases intracellular Mg, which could be associated with chronic sleep deprivation-induced cardiovascular events. PMID- 15754838 TI - Assessment of autonomic activity during daily life of patients with head-up tilt induced prolonged asystole. AB - Autonomic activity is important in the pathophysiology of neurally mediated syncope (NMS) patients diagnosed by the head-up tilt (HUT) test, and prolonged systole induced by HUT is associated with a malignant vasovagal syndrome. This study investigated whether or not daily autonomic activity evaluated by heart rate variability (HRV) assessed through 24-hour ambulatory ECG monitoring was augumented in HUT-induced asystole (AS; > 30 s) and whether or not HRV differed between NMS patients with and without AS. Ambulatory ECG monitoring was performed in 176 patients with suspected NMS and in 62 age-matched healthy control subjects (C group, age: 36 +/- 13 yr), with HRV time and frequency domain indices measured within 48 hours of HUT. Six patients displayed prolonged AS (PAS group, age: 33 +/- 12 yr), and 109 patients had positive HUT (P group, age: 29 +/- 17 yr), while 61 patients had negative HUT (N group, age: 40 +/- 18 yr). Several HRV indices in the P group were higher than those in the N and C groups. However, HRV indices did not differ between the P group and the PAS group. It is concluded that impairment of autonomic activity specifically related to orthostatic stress could be important in the pathophysiology of HUT-induced prolonged asystole, while impairment of autonomic activity in daily life is not as important in NMS patients with HUT-induced prolonged asystole. PMID- 15754839 TI - Positive impact of social intervention on disturbed neurobehavioral function in an elderly community-dwelling population: longitudinal investigation for longevity and aging in Hokkaido County (LILAC). AB - Several kinds of health consultation and rehabilitation for functional disorders aimed at stroke prevention and maintenance of cognitive function in an elderly population in Hokkaido county, Japan. Changes in cardiovascular and neurobehavioral endpoints between 2000 and 2002 were assessed in 72 of 115 subjects over 75 years of age. Direct social intervention, including lifestyle modification can have a positive impact, notably on subjects with cardiovascular disorders. PMID- 15754840 TI - Chronomic community screening reveals about 31% depression, elevated blood pressure and infradian vascular rhythm alteration. AB - Depression, which is a risk factor for cardiac morbidity and mortality, is not an unusual occurrence among individuals with coronary heart disease (CHD), but evidence concerning its role in the pathogenesis of this condition is less clear. Ambulatory blood pressure monitoring (ABPM) has become an important tool in the diagnosis and management of hypertension. Several previous studies have indicated that various kinds of target organ damage and cardiovascular morbidity are more strongly associated with a diagnosis by ABPM than through spot-checks in a clinical setting. This study investigated whether depressive mood was associated with changes in the about-weekly (circaseptan) and half-weekly (circasemiseptan) variations in blood pressure (BP) and heart rate (HR), including a BP surge on Mondays, in community-dwelling subjects monitored chronomically for the time structure (chronome) of their BP and HR variabilities. From April 2001 to April 2003, 217 subjects (85 men and 132 women; mean age: 56.8 +/- 11.3 yr) from U town, Hokkaido (latitude: 43.45 degrees N, longitude: 141.85 degrees E), self monitored their BP and HR for 7 days starting around 11 a.m. on Thursday, and took readings at 30-minute intervals between 7 a.m. and 10 p.m., then at 60 minute intervals between 10 p.m. and 7 a.m. The data were retrieved and analyzed on a PC with appropriate commercial software (TM-2430-15; A&D Co., Japan). Subjects were asked about 15 items on a depression rating scale through a self administered questionnaire. When the score amounted to 5 or higher, subjects were considered to be depressive. Student's t-test, a one-way analysis of variance (ANOVA), and cosinor methods with parametric tests were also used. A p-value below 0.05 was considered to indicate statistical significance (below 0.10: borderline statistical significance). Depression rating scales were obtained for 192 out of the 217 subjects enrolled in this study. Depression scores were (>) 5 in 72 subjects. The average values of systolic (S) and diastolic (D) BP were statistically significantly higher in depressed subjects (SBP: 129.2 vs 124.5 mmHg; p = 0.034; DBP: 79.0 vs 76.5 mmHg; p = 0.041). The 7-day average for HR did not differ between subjects with depression scores of < 5 or > 5. DBP dipping was less in the depressed subjects (16.30 vs 18.22%; p = 0.048). The dipping ratios of SBP and HR showed no statistically significant difference. In the group with depression scores of < 5, HR variability (estimated by the SD of HR and HR dip) was higher during vacations and lower on Mondays. The 24-h BP measures showed a novelty effect and a surge on Mondays. In the depressed group, a prominent circaseptan rhythm appeared to replace the novelty effect, vacation dip, and Monday surge. The results of this investigation indicate the clinical importance of the monitoring of depressed subjects. Fewer than 7 days of monitoring means a greater risk of false diagnosis, and thus a therapeutic decision including potentially unnecessary or inappropriate long-term treatment. Records shorter than 7 days would not have detected circaseptan BP dysrhythmia associated with a depressive state. Prominent circaseptans can provide new indications on the mechanisms underlying the strong relation between depression and adverse cardiac events. Future studies should aim at determining whether the treatment of depression, especially from the standpoint of a chronodiagnosis and chronotherapy, can reduce the incidence of adverse cardiac events, and whether this depends upon restoring normal BP and HR variability, i.e. anormal BP and HR chronome. PMID- 15754841 TI - Can brain dysfunction be a predisposing factor for metabolic syndrome? AB - The various mechanisms that may explain the association between brain dysfunction and the pathogenesis of metabolic syndrome (MS) leading to cardiovascular disease and type 2 diabetes have been reviewed. A Medline search was conducted until September 2003, and articles published in various national and international journals were reviewed. Experts working in the field were also consulted. Compelling evidence was found that saturated and total fat and low dietary n-3 fatty acids and other long-chain polyunsaturated fatty acids (PUFAs) in conjunction with sedentary behavior and mental stress combined with various personality traits can enhance sympathetic activity and increase the secretion of catecholamine, cortisol and serotonin, all of which appear to be underlying mechanisms involved in MS. Excess secretion of these neurotransmitters in conjunction with underlying long-chain PUFA deficiency may damage the neurons in the ventromedial hypothalamus and insulin receptors in the brain, in particular during fetal life, infancy and childhood, and lead to their dysfunction. Since 30 50% of the fatty acids in the brain are long-chain PUFAs, especially omega-3 fatty acids which are incorporated in the cell membrane phospholipids, it is possible that their supplementation may have a protective effect. Omega-3 fatty acids are also known to enhance parasympathetic activity and to increase the secretion of anti-inflammatory cytokines as well as acetylecholine in the hippocampus. It is possible that a marginal deficiency of long-chain PUFAs, especially n-3 fatty acids, due to poor dietary intake during the critical period of brain growth and development in the fetus, and later in the infant and also possibly in the child, adolescent and adult may enhance the release of tumor necrosis factor-alpha (TNF-alpha) interleukin (IL)-1, 2 and 6 and cause neuronal dysfunction. Experimental studies indicate that ventromedial hypothalamic lesions in rats induce hyperphagia, resulting in glucose intolerance and insulin resistance. Treatment with neuropeptide Y abolished hyperphagia and ob mRNA (leptin mRNA) in this animal model. Long-term infusion of norepinephrine and serotonin into the ventromedial hypothalamus impaired pancreatic islet function inasmuch as ventromedial hypothalamic norepinephrine and serotonin levels were elevated in hyperinsulinemic and insulin-resistant animals. Treatment with insulin was associated with restoration of hypothalamic neurotransmitter abnormalities, indicating that ventromedial hypothalamus dysfunction can impair pancreatic beta cells resulting in metabolic abnormalities consistent with MS. Treatment with omega-3 fatty acids, beta blockers, ACE inhibitors, estrogen, and meditation may have a beneficial effect on insulin receptors and ventromedial hypothalamic dysfunction. However, no definite or precise insight into the pathophysiological link between MS, brain function and nutrition is available. Despite this, epidemiological studies and intervention trials indicate that treatment with n-3 fatty acids may be adopted in clinical practice and used to direct therapy for prevention of type 2 diabetes, hypertension, coronary artery disease (CAD), and atherosclerosis, thereby indicating that MS may also respond to this treatment. PMID- 15754842 TI - 100 or 30 years after Janeway or Bartter, Healthwatch helps avoid 'flying blind'. AB - Longitudinal records of blood pressure (BP) and heart rate (HR) around the clock for days, weeks, months, years, and even decades obtained by manual self measurements (during waking) and/or automatically by ambulatory monitoring reveal, in addition to well-known large within-day variation, also considerable day-to-day variability in most people, whether normotensive or hypertensive. As a first step, the circadian rhythm is considered along with gender differences and changes as a function of age to derive time-specified reference values (chronodesms), while reference values accumulate to also account for the circaseptan variation. Chronodesms serve for the interpretation of single measurements and of circadian and other rhythm parameters. Refined diagnoses can thus be obtained, namely MESOR-hypertension when the chronome-adjusted mean value (MESOR) of BP is above the upper limit of acceptability, excessive pulse pressure (EPP) when the difference in MESOR between the systolic (S) and diastolic (D) BP is too large, CHAT (circadian hyper-amplitude tension) when the circadian BP amplitude is excessive, DHRV (decreased heart rate variability) when the standard deviation (SD) of HR is below the acceptable range, and/or ecphasia when the overall high values recurring each day occur at an odd time (a condition also contributing to the risk associated with 'non-dipping'). A non-parametric approach consisting of a computer comparison of the subject's profile with the time-varying limits of acceptability further serves as a guide to optimize the efficacy of any needed treatment by timing its administration (chronotherapy) and selecting a treatment schedule best suited to normalize abnormal patterns in BP and/or HR. The merit of the proposed chronobiological approach to BP screening, diagnosis and therapy (chronotheranostics) is assessed in the light of outcome studies. Elevated risk associated with abnormal patterns of BP and/or HR variability, even when most if not all measurements lie within the range of acceptable values, becomes amenable to treatment as a critical step toward prevention (prehabilitation) to reduce the need for rehabilitation (the latter often after costly surgical intervention). PMID- 15754843 TI - Circannual and/or transannual variations in growth hormone and beyond--call for long data series. AB - A meta-analysis of growth hormone data showing similar double amplitudes of 36 and 34% of the mean from the fit of cosine curves with periods of 1.3 and 1.0 year, respectively, suggests the desirability of analyzing available time series on variables of interest in biomedicine, to explore heretofore unknown longer than-yearly cycles that may interact (beat) with yearly ones. PMID- 15754845 TI - Brachio-ankle pulse wave velocity and cardio-ankle vascular index (CAVI). AB - In order to diagnose arteriosclerosis in any part of the body, pulse wave velocity (PWV) measurement is a useful approach. However, it is considered that the technique of PWV measurement should be simplified. A new method for measuring PWV has therefore been proposed in Japan. The PWV of the brachial artery (ba) and the ankle was measured by applying air pressure with the aid of a volume plethysmograph. Comparisons between the baPWV measurement method and the conventional method are currently being performed. Since satisfactory results have been obtained to date, baPWV has gained popularity throughout Japan. Since this method measures PWV in the arm and foot, it may be said that aortic PWV is not reflected though a large amount of past PWV measurements. BaPWV is influenced by blood pressure. With the baPWV technique, blood pressure compensation is not carried out. Furthermore, the pulse pressure is measured by air pressure; therefore any stimulus that exerts pressure on an artery may influence these results. Due to these reasons, a cardie-ankle vascular index (CAVI) has been proposed in which the pressure wave form indicating the closing of the aortic valve appears in the form of an arterial pressure wave after a fixed delay time. This delay is the time difference between the actual closing of the aortic valve and the measuring point. Prior to the introduction of baPWV, PWV was measured in the carotid artery and foot. As in traditional PWV, baPWV uses the delay time, but between the brachial artery and the ankle artery. However, the carotid artery differs from the brachial artery, and the measured value differs depending on whether the arteriosclerosis is present in the carotid artery or the brachial artery. CAVI is calculated from the ECG, PCG, brachial artery waveform and ankle artery waveform using a special algorithm. This new method represents a breakthrough in the diagnosis of atherosclerosis. PMID- 15754844 TI - A Japanese-Russian collaborative study on aging and atherosclerosis. AB - In Russia, the average life span is decreasing. This phenomenon has not been previously observed in the western hemisphere during periods of relative peace. Although this trend can be attributed to various causes, the available statistics suggest that approximately 55% of the total mortality in Russia at the present time is due to cardiovascular events. Similarly, cardiovascular events are also increasing in Japan and are associated with changes in eating habits. A Japanese Russian international collaborative study was therefore initiated to investigate this. Data on 102 normal Russian subjects were obtained from a total of 345 patients treated at the Smolensk State Medical Academy outpatients' clinic. The data on the Japanese patients were collected at Tohoku University, Tohoku Rosai Hospital, and Fukuda Denshi Company. Sixty-seven healthy male Japanese volunteers were compared with 44 healthy male Russian patients. In this study, Vasera VS1000 (Fukuda Denshi, Tokyo) was utilized to measure pulse wave velocity (PWV). Linear regression analysis was performed, and the results showed that brachial-ankle PWV (baPWV) was significantly higher in the Russian compared with the Japanese study group. With regard to aging, PWV was found to increase both in the Japanese and Russian subjects, with a significant increase in the latter group during their 40s compared with the Japanese, and with this tendency being carried forward into the 50s. This may be the principal cause for the higher incidence of cardiovascular events reported among Russians. Furthermore, when a straight line regression analysis was performed, the zero axis intercept in both cases showed almost the same value. These results indicated that the PWV may be almost the same in Japanese and Russian populations at the time of birth. In conclusion, at the time of birth, there is no difference in the atherosclerotic status of Japanese and Russian infants. However, over time atherosclerosis becomes more apparent and progresses significantly in the case of Russian subjects. This finding is observed through the analysis of PWV. The present authors intend to continue their study aimed at preventing the further development of atherosclerosis through medical intervention. PMID- 15754846 TI - Circadian hypo- and hyper-amplitude-tension (CHAT) associated with putative pheochromocytoma and 12-hourly phenoxybenzamine treatment. AB - A 26-year-old white woman had an ablated sino-atrial node and ventricular pacemaker as an unusual feature of a pheochromocytoma-compatible history. Her status quo included, on three occasions, elevated 24-hour urinary epinephrine and metanephrine excretion. She monitored her blood pressure (BP) and heart rate (HR) at 15- to 60-minute intervals over several days, with interruptions, before and after the institution of 10 mg phenoxybenzamine per os every 12 hours (between 7:30 and 8:00 and between 19:30 and 20:00), with continued monitoring over several months. Her data were summarized for consecutive 3-day intervals by sphygmochron. Circadian parameters and original data are compared with gender- and age-specified reference values, yielding also non-parametric endpoints, such as the percentage time elevation, the extent of excess, and the timing of excess, that all can be acceptable for some days but unacceptable for other days. In her broader time structure, or chronome, cosinor analyses revealed a prominent and statistically significant circadian rhythm in BP and HR before and during the 12 hourly therapy. The 12-hour component of BP was more prominent during therapy than prior to it. A statistically significant decreasing trend occurred before therapy, and recurred during treatment. Chronomically interpreted monitoring revealed: 1) the persistence of a statistically significant circadian rhythm during 12-hourly phenoxybenzamine treatment; 2) days-long changes in BP MESOR, the duration of which could not be previously determined based on spotchecks; 3) changes in the circadian amplitude of BP, which can be either very small or very large, compatible with the diagnosis of intermittent circadian hyper-amplitude tension (CHAT); and 4) a very wide range of BP and HR values, so that occasional (casual) measurements fail to convey the dynamics that may underlie this infrequently found clinical condition of an elevated catecholamine excretion compatible with a pheochromocytoma. All findings support the need for long-term monitoring of BP and HR that may account for controversy in earlier publications. PMID- 15754848 TI - Mechanisms of acute myocardial infarction study (MAMIS). AB - Acute myocardial infarction (AMI) is a highly dynamic event, which is associated with marked neuroendocrinological dysfunction in addition to cardiac damage. The immediate trigger for AMI is not precisely known. Studies conducted by Lown, Braunwald, Halberg, Otsuka and our group have demonstrated a marked increase in sympathetic activity, oxidative stress, and magnesium and potassium deficiency during AMI. Clinical studies have reported an increased incidence of AMI, sudden death and ischemia during first quarter of the day when there is a rapid withdrawal of vagal activity and increase in sympathetic tone. In one case control study of 202 patients with AMI, there was a significant (P < 0.02) increase in cardiac events in the second quarter of the day compared to other quarters, respectively (16.8%, 41.0%, 13.8%, 28.2% per quarter). This characteristic remained prevalent in both men and women and among patients with and without known AMI (n = 52), diabetes (n = 53) or hypertension (n = 75). Triggers of AMI were noted among 162 (82.2%) of the patients. Neuropsychological mechanisms were observed as follows: emotional stress (45.5%), sleep deprivation (27.7%), cold climate (29.2%), hot climate (24.7%), large meals (47.5%) and physical exertion (31.2%). These triggering factors are known to enhance sympathetic activity and decrease vagal tone, resulting in an increased secretion of plasma cortisol, noradrenaline, aldosterone, angiotension-converting enzyme (ACE), interleukin (IL)-1, -2, -6, -18, and tumor necrosis factor-alpha (TNF alpha), all of which are are proinflammatory agents. There is also a deficiency in the serum levels of vitamin A, E, and C and magnesium, potassium, melatonin, and IL-10 (an anti-inflammatory agent). In our study, we found a decrease in magnesium, potassium, vitamin A, E, C and beta carotene combined with an increase in thiobarbituric acid-reactive substances (TBARS), MDA and diene conjugates, TNF alpha and IL-6, all of which are indicators of oxidative damage and proinflammatory activity, respectively. PMID- 15754847 TI - No baseline for blood pressure in ordinary life: another case of transient chronome alterations. AB - Half-hourly systolic (S) and diastolic (D) blood pressure (BP) and heart rate (HR) monitoring with gaps demonstrates transient elevations of unknown origin of the chronome (time structure)-adjusted mean, the MESOR, M, of BP, and of the circadian double amplitude of BP of TS, a clinically healthy-appearing engineering student who was 27 years of age at the start of the study. An assessment of large and small arterial vessel elasticity was in keeping with no detection of functional or structural alteration in her vasculature. The recovery of normality in BP endpoints at the time of this report is speculatively associated with a weight loss of 40 pounds and the formation of a friendship by correspondence. Whatever the underlying mechanisms of a long series of abnormal records may be, a 'baseline' can include weeklong spans of abnormality and is best replaced by comparisons with chronomic reference values. PMID- 15754849 TI - Increased concentrations of lipoprotein(a), circadian rhythms and metabolic reactions evoked by acute myocardial infarction, associated with acute reactions in relation to large breakfasts. AB - Of 54 patients with acute coronary artery disease (CAD) that were included in this study, 41 patients had acute myocardial infarction (AMI), five patients possible MI, four patients unstable angina and the remaining four angina pectoris. The control subjects (n = 85) were randomly selected from the general population of the city of Moradabad of similar age range after exclusion for CAD (n = 9), diabetes (n = 6) and excess intake of trans fatty acids (n = 20). The incidence of lipoprotein(a) excess (> 30 mg/dl; 42.6 vs 24.7%; P < 0.05) and mean concentration of lipoprotein(a) (Lp[a], 6.4 mg/dl, 95% confidence interval: 2.8 10.5; P < 0.05) was significantly greater in the acute CAD group compared with control subjects. Mean total cholesterol and triglycerides were significantly higher and mean nitrite level lower in the study group as compared with the control group.There was a significant greater incidence of cardiac events in the second quarter of the day compared with the fourth quarter. Lp(a), triglycerides, blood glucose, plasma insulin, malondialdehyde, diene conjugates, TBARS and TNF alpha and IL-6 levels, which were significantly greater during the acute phase, showed a significant decline and serum nitrite and coenzyme Q demonstrated an increase at 4 weeks of follow-up when the acute reactions evoked by MI had been controlled. Large breakfasts were a predisposing factor for cardiac events in the second quarter of the day and it was significantly associated with metabolic reactions. The findings indicate that acute reactions as a result or as circadian rhythms appear to be important in the pathogenesis of AMI-associated complications and that a large breakfast in association with nitrite deficiency may further trigger the circadian rhythms. However, more studies in a larger number of subjects would be necessary in order to confirm our findings. PMID- 15754850 TI - Day-to-day variability of the long-term fractal component of heart rate variability (beta) and the effect of an antianginal agent in patients with silent myocardial ischemia. AB - The deterioration of the long-term fractal component of heart rate variability (beta) is reported in patients with organic heart disease. To examine the day-to day variability of beta and to study the effects of nipradilol (NIP), an antianginal agent, on beta in patients with coronary artery disease (CAD), we performed 24-hour ambulatory ECG (AECG) in 24 patients with silent myocardial ischemia (SMI) and obtained the conventional frequency domain heart rate variability (HRV) indices and beta using the MemCalc system. Patients were divided into two groups: group I consisted of 10 patients (64 +/- 5 years of age) for the assessment of the day-to-day variability of beta; group II comprised 14 patients (60 +/- 6 years of age) for evaluation of the effect of NIP (3 mg b.i.d). In group I, AECG was repeated twice a week, while AECG was recorded before NIP treatment (placebo) and on the last day of the 4-week NIP treatment in group II. In group I, there was significant correlation between the values of beta on day 1 and day 2 (r = 0.79, p < 0.05), the difference being 0.09 +/- 0.15 (coefficient of variance: 6.8%). Bland and Altman's analysis indicated that the results were easily reproducible, making this approach feasible for clinical use. In group II, NIP significantly decreased the values of beta by 10.6% (from -1.23 +/- 0.09 to -1.18 +/- 0.07; p < 0.05). NIP also significantly increased the conventional HRV indices. The degree of decrement of beta by NIP was larger than that of the day-to-day variability. In conclusion, a day-to-day variability exists in beta in patients with SMI, but its value is relatively small. A daily dose of 6 mg NIP ameliorates beta and overcomes this variability. PMID- 15754851 TI - Melatonin, refractory hypertension, myocardial ischemia and other challenges in nightly blood pressure lowering. AB - A test of the relative merits of timed melatonin for the treatment of cardiac ischemia as well as hypertension refractory to other drugs is documented against the background of earlier chronobiological studies on blood pressure (BP), disease risks, circadian hyper-amplitude-tension and melatonin effects broadly. PMID- 15754853 TI - About-weekly variations in nocturia. AB - The aim of this study was to assess components of variation in nocturia and to determine any putative geomagnetic influence. A 54-year old man with benign prostatic hyperplasia had recorded for about 4 years the number of times he awoke each night to urinate. The data have been reanalyzed for chronomics, the mapping of time structures (chronomes), involving the computation of least squares spectra of the urinary record and of environmental variables recorded during the same 4-year span. In addition to the previously reported monthly variation, other periodicities have been documented, including two separate components with periods of one week and of a near-week. The precise 7-day period may be a mainly exogenous resonance with external influences such as a weekly social schedule, whereas the near-week may be a partial resonance with natural changes in geomagnetics, reflecting in part changes in other non-photic natural environmental factors. PMID- 15754852 TI - Time structures, chronomes, of soldiers' stature mimicking Hale cycle in neonatal body length. AB - The systematic patterns in human adult physical stature are explored in connection with Wolf's relative sunspot numbers. This topic should be of interest to economists, physicians as well as physicists. There is a need for more than a check of any similarity of curves in variables approximating the economy and human stature and for more than the application of mathematical models, as done herein. Ours is at best a halting step at one frequency, presented only to document the challenge of a transdisciplinary approach to multifrequency intermodulations of hosts of variables, yet to be untangled. The circumstance that at birth some decisions concerning adult stature are already made is challenging. The signature of the environment in terms of the about 21-year Hale bipolarity cycle of Wolf's relative sunspot numbers found in adult soldiers shows that the association at birth is not a transient one, even if other evidence beyond our scope herein points to the possibility that observations on presumably healthy soldiers can be extrapolated to abnormal growth. Nonetheless, the task for those concerned with short stature could become preventive if the sensitive stages when growth may be inhibited by the environment could be found, as well as means to shield from or counter the undesirable effects. PMID- 15754854 TI - Addition of rhythm to non-pulsatile circulation. AB - The development of a rotary blood pump (RP) is desirable as it can be used as a small ventricular assistance device (VAD). However, a RP does not generate any pulse. It may be physiologically better for the patient if the RP could generate a pulse. We have attempted to develop a device that produces a pulse in the RP. Intra-aortic balloon pumping (IABP) is effective in producing a pulse. However, the IABP cannot be implanted inside the body. Therefore, an attempt was made to develop pulse-generating equipment that was not driven by air pressure. The ball screw motor was considered as a possible candidate. In the future, we plan to apply small shape memory alloys. An electrohydraulic system was adopted, and actuator power output was connected to the diaphragm. The diaphragm was placed outside the ventricle. Most RPs developed throughout the world drain blood from the ventricle. The pulse wave should be generated if a pulse is added by the part from which blood is being drawn. In this study, animal experiments were conducted and the output assistance was tested from outside the ventricle. The device operated effectively in the animal experiment. The RP can easily be equipped with this device at the time of performing the implant operation. For a patient with problems of peripheral circulation and the internal organ function, it may prove to be an effective device. PMID- 15754856 TI - Emotional intelligence skills for maintaining social networks in healthcare organizations. AB - For healthcare organizations to survive in these increasingly challenging times, leadership and management must face mounting interpersonal concerns. The authors present the boundaries of internal and external social networks with respect to leadership and managerial functions: Social networks within the organization are stretched by reductions in available resources and structural ambiguity, whereas external social networks are stressed by interorganizational competitive pressures. The authors present the development of emotional intelligence skills in employees as a strategic training objective that can strengthen the internal and external social networks of healthcare organizations. The authors delineate the unique functions of leadership and management with respect to the application of emotional intelligence skills and discuss training and future research implications for emotional intelligence skill sets and social networks. PMID- 15754858 TI - Assessment of management education and training for healthcare providers in the Slovak Republic. AB - The authors' purpose of this study was to analyze the current system of healthcare managers' education in the Slovak Republic. The study was essentially qualitative and used a triangular approach. This included an analysis of the range of healthcare management education providers, an assessment of needs that was based on questionnaires distributed to a sample of top managers in hospitals, and a focus group discussion. The findings revealed a gap between the estimated needs and the current capacities for management education. The results serve as a baseline for a discussion of future management training and education for healthcare providers in the Slovak Republic. PMID- 15754857 TI - Contrasting the American College of Healthcare Executives' code of ethics with undergraduate health administration students' values and ethical decision choices. AB - Although administrative ethics are imbedded into the code of ethics of the American College of Healthcare Executives (ACHE), understanding the values and ethical decision-making practices of health administration students can help shape content and curriculum for health administration programs in the future. The study surveyed a sample of undergraduate health administration students to examine their sense of honesty and ethical decision-making practices. The sampled students completed the Comparative Emphasis Scale, which measured the student's sense of honesty, fairness, and integrity, and 10 short cases of administrative ethical issues derived from the ACHE Code of Ethics. The findings from the study indicated that the health administrative students had moderate to high mean scores on the ethical scales measuring achievement (15.86), concern for others (17.82), fairness (17.67), and honesty (18.21). The students' overall mean score for the 10 ethical cases was 3.51 on a 5-point scale, with 1 indicating a low likelihood and 5 a high likelihood. Pearson's product-moment correlation coefficient showed a minimum relationship between results of the Comparative Emphasis Scale and responses to ethical decision-making cases, and it showed no significant relationship between race, gender, and class (freshman, sophomore, junior, or senior) of the students. The results of the study have indicated that the sampled undergraduate health administration students respond at a moderate to high ethical level to this standardized scale and cases involving ACHE administrative ethical issues. Future research should explore the relationship between select variables concerning employment history, industry, position, and demographics characteristics in people's ethical choices. PMID- 15754855 TI - Chronoastrobiology: proposal, nine conferences, heliogeomagnetics, transyears, near-weeks, near-decades, phylogenetic and ontogenetic memories. AB - "Chronoastrobiology: are we at the threshold of a new science? Is there a critical mass for scientific research?" A simple photograph of the planet earth from outer space was one of the greatest contributions of space exploration. It drove home in a glance that human survival depends upon the wobbly dynamics in a thin and fragile skin of water and gas that covers a small globe in a mostly cold and vast universe. This image raised the stakes in understanding our place in that universe, in finding out where we came from and in choosing a path for survival. Since that landmark photograph was taken, new astronomical and biomedical information and growing computer power have been revealing that organic life, including human life, is and has been connected to invisible (non photic) forces, in that vast universe in some surprising ways. Every cell in our body is bathed in an external and internal environment of fluctuating magnetism. It is becoming clear that the fluctuations are primarily caused by an intimate and systematic interplay between forces within the bowels of the earth--which the great physician and father of magnetism William Gilbert called a 'small magnet'- and the thermonuclear turbulence within the sun, an enormously larger magnet than the earth, acting upon organisms, which are minuscule magnets. It follows and is also increasingly apparent that these external fluctuations in magnetic fields can affect virtually every circuit in the biological machinery to a lesser or greater degree, depending both on the particular biological system and on the particular properties of the magnetic fluctuations. The development of high technology instruments and computer power, already used to visualize the human heart and brain, is furthermore making it obvious that there is a statistically predictable time structure to the fluctuations in the sun's thermonuclear turbulence and thus to its magnetic interactions with the earth's own magnetic field and hence a time structure to the magnetic fields in organisms. Likewise in humans, and in at least those other species that have been studied, computer power has enabled us to discover statistically defined endogenous physiological rhythms and further direct effects that are associated with these invisible geo- and heliomagnetic cycles. Thus, what once might have been dismissed as noise in both magnetic and physiological data does in fact have structure. And we may be at the threshold of understanding the biological and medical meaning and consequences of these patterns and biological-astronomical linkages as well. Structures in time are called chronomes; their mapping in us and around us is called chronomics. The scientific study of chronomes is chronobiology. And the scientific study of all aspects of biology related to the cosmos has been called astrobiology. Hence we may dub the new study of time structures in biology with regard to influences from cosmo- helio- and geomagnetic rhythms chronoastrobiology. It has, of course, been understood for centuries that the movements of the earth in relation to the sun produce seasonal and daily cycles in light energy and that these have had profound effects on the evolution of life. It is now emerging that rhythmic events generated from within the sun itself, as a large turbulent magnet in its own right, can have direct effects upon life on earth. Moreover, comparative studies of diverse species indicate that there have also been ancient evolutionary effects shaping the endogenous chronomic physiological characteristics of life. Thus the rhythms of the sun can affect us not only directly, but also indirectly through the chronomic patterns that solar magnetic rhythms have created within our physiology in the remote past. For example, we can document the direct exogenous effects of given specific solar wind events upon human blood pressure and heart rate. We also have evidence of endogenous internal rhythms in blood pressure and heart rate that are close to but not identical to the period length of rhythms in the solar wind. These were installed genetically by natural selection at some time in the distant geological past. This interpretive model of the data makes the prediction that the internal and external influences on heart rate and blood pressure can reinforce or cancel each other out at different times. A study of extensive clinical and physiological data shows that the interpretive model is robust and that internal and external effects are indeed augmentative at a statistically significant level. Chronoastrobiological studies are contributing to basic science--that is, our understanding is being expanded as we recognize heretofore unelaborated linkages of life to the complex dynamics of the sun, and even to heretofore unelaborated evolutionary phenomena. Once, one might have thought of solar storms as mere transient 'perturbations' to biology, with no lasting importance. Now we are on the brink of understanding that solar turbulences have played a role in shaping endogenous physiological chronomes. There is even documentation for correlations between solar magnetic cycles and psychological swings, eras of belligerence and of certain expressions of sacred or religious feelings. Chronoastrobiology can surely contribute to practical applications as well as to basic science. It can help develop refinements in our ability to live safely in outer space, where for example at the distance of the moon the magnetic influences of the sun will have an effect upon humans unshielded by the earth's native magnetic field. We should be better able to understand these influences as physiological and mechanical challenges, and to improve our estimations of the effects of exposure. Chronoastrobiology moreover holds great promise in broadening our perspectives and powers in medicine and public health right here upon the surface of the earth. Even the potential relevance of chronoastrobiology for practical environmental and agricultural challenges cannot be ruled out at this early stage in our understanding of the apparently ubiquitous effects of magnetism and hence perhaps of solar magnetism on life. The evidence already mentioned that fluctuations in solar magnetism can influence gross clinical phenomena such as rates of strokes and heart attacks, and related cardiovascular variables such as blood pressure and heart rate, should illustrate the point that the door is open to broad studies of clinical implications. The medical value of better understanding magnetic fluctuations as sources of variability in human physiology falls into several categories: 1) The design of improved analytical and experimental controls in medical research. Epidemiological analyses require that the multiple sources causing variability in physiological functions and clinical phenomena be identified and understood as thoroughly as possible, in order to estimate systematic alterations of any one variable. 2) Preventive medicine and the individual patients'care. There are no flat 'baselines', only reference chronomes. Magnetic fluctuations can be shown statistically to exacerbate health problems in some cases. The next step should be to determine whether vulnerable individuals can be identified by individual monitoring. Such vulnerable patients may then discover that they have the option to avoid circumstances associated with anxiety during solar storms, and/or pay special attention to their medication or other treatments. Prehabilitation by self-help can hopefully complement and eventually replace much costly rehabilitation. 3) Basic understanding of human physiological mechanisms. The chronomic organization of physiology implies a much more subtle dynamic integration of functions than is generally appreciated. All three categories of medical value in turn pertain to the challenges for space science of exploring and colonizing the solar system. The earth's native magnetic field acts like an enormous umbrella that offers considerable protection on the surface from harsh solar winds of charged particles and magnetic fluxes. The umbrella becomes weaker with distance from the earth and will offer little protection for humans, other animals, and plants in colonies on the surface of the moon or beyond. Thus it is important before more distant colonization is planned or implemented to better understand those magnetism-related biological- solar interactions that now can be studied conveniently on earth. (ABSTRACT TRUNCATED) PMID- 15754859 TI - Real-time patient satisfaction survey and improvement process. AB - The purpose of this article is to describe how one multidisciplinary hospital responded to patient-satisfaction issues and improved communication throughout its organization by implementing a real-time assessment of patient and staff satisfaction for a faster and better-focused improvement process. The survey process is based on eliciting information from several different sources in a manner that allows corrective action plans to be made and implemented within 4 to 8 weeks of patient encounters. Organized groups can then review feedback from the implemented action plans within 9 to 16 weeks of patient encounters. This 4-month process is repeated on a quarterly basis, as lessons learned from the previous cycle are fed into the upcoming survey process for continuous patient satisfaction improvement. Employees, faculty, and administrators have accepted the Real-Time Patient Satisfaction Survey and Improvement Process as a routine activity within the normal operating structure at the Moffitt Cancer Center. This activity of problem identification-action-feedback has been well integrated in the system and will continue to rotate throughout all patient care clinical services at the Moffitt Cancer Center. The program has become a method for goal setting and establishing management accountability. As an adaptation of continuous quality improvement, The Real-Time Patient Satisfaction Survey and Improvement Process at the Moffitt Cancer Center is applicable for use in other hospitals and cancer centers in the United States. The general design, materials, and analysis plan can be directed toward the needs of the specific institution (and are available for distribution by contacting the authors). PMID- 15754860 TI - The "new" medical malpractice crisis--part 1. AB - It is important to put the current medical malpractice crisis into the historical context of the past several decades. Doing so provides an important perspective from which to understand the current iteration. One may reasonably conclude that the present medical malpractice situation is only the latest outbreak of a continuing, chronic condition, rather than a distinct, unusual event. In this regard, it is analogous to a chronic disease that occasionally flares up. Chronicity suggests the presence of major underlying problems, which may be linked to insurance carriers' business cycles as much as reflecting increases in either medical malpractice or the numbers and value of claims. It is useful to bear in mind that the fact of a claim may or may not indicate actual medical malpractice, and increased claims could well correlate more closely with patients' disgruntlement with the medical delivery system, access to it, and the way they were treated by staff than with significant injuries for which compensation should be paid. Regulatory and public policy efforts to date have only affected the problem of medical malpractice at the margin. Apparently, the core of the problem has not been addressed; in fact, it seems as yet to be unidentified. Solutions that focus on the economic dimensions only address the symptoms-claims for medical malpractice-and apparently have done nothing to correct the root cause(s). Part 2 of this two-part series considers and analyzes the current medical malpractice insurance crisis. Its evolution and analysis of specific aspects may provide guidance in understanding how to predict its future course. More important, the analysis will suggest guidance as to how organizations may reduce the potential for the problem and protect themselves from the negative aspects, should it occur. PMID- 15754861 TI - Second medical opinions sought by patients. PMID- 15754862 TI - The body's story: a case report of hypnosis and physiological narration of trauma. AB - Adult Posttraumatic Stress Disorder secondary to childhood sexual abuse is clinically complicated by its increasingly noted deficient linguistic recording of the abuse, perhaps partially explaining consequent difficulties with verbalizing in therapy. A single case illustrates that hypnotically utilizing the body-emotion register of encrypted sexual abuse trauma may not only afford more naturalistic retrieval and purgation of the experience, but may also provide the very medium for the healing narrative required for recovery. The patient's original and continuing therapist was also present as support and observer for all but 1 of 25 hypnosis sessions. Treatment gains were robust at 3-year follow up. This case suggests that effective treatment for sexual abuse PTSD may in some instances reside in more nonverbally sensitive interventions not aiming to prove, probe, or process linguistic reconstructions of memory. This is the first published report of such a bodily narrative in hypnosis. PMID- 15754863 TI - Hypnosis and irritable bowel syndrome: a review of efficacy and mechanism of action. AB - Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder characterized by abdominal pain, distension, and an altered bowel habit for which no cause can be found. Despite its prevalence, there remains a significant lack of efficacious medical treatments for IBS to date. In this paper we reviewed a total of 14 published studies (N=644) on the efficacy of hypnosis in treating IBS (8 with no control group and 6 with a control group). We concluded that hypnosis consistently produces significant results and improves the cardinal symptoms of IBS in the majority of patients, as well as positively affecting non-colonic symptoms. When evaluated according to the efficacy guidelines of the Clinical Psychology Division of American Psychological Association, the use of hypnosis with IBS qualifies for the highest level of acceptance as being both efficacious and specific. In reviewing the research on the mechanism of action as to how hypnosis works to reduce symptoms of IBS, some evidence was found to support both physiological and psychological mechanisms of action. PMID- 15754864 TI - Utilizing hypnosis and ego-state therapy to facilitate healthy adaptive differentiation in the treatment of sexual disorders. AB - Much of the literature focuses on the pathology that falls to the far right of the Watkins (1997) differentiation-dissociation continuum, such as Dissociative Identity Disorder and Dissociative Disorder NOS. Adding a "far left" to this continuum, as well as a construct of what the "far left" looks like, makes apparent the value of healthy adaptive differentiation for those individuals that fall to the "far left" of the spectrum; those who don't differentiate enough. A discussion of sexual dysfunction at this end of the continuum and cases of Hypoactive Sexual Desire Disorder and Vaginismus demonstrate the clinical effectiveness of an approach combining hypnosis and ego-state therapy to facilitate healthy adaptive differentiation. PMID- 15754865 TI - Three failures of direct suggestion in psychogenic dermatitis followed by successful intervention. AB - Three adult cases of psychogenic dermatitis of atypical presentation were treated with direct suggestion under hypnosis (DSUH), which included suggestions for developing cooling, soothing and healing numbness in the affected areas. After a trial of 5 sessions over a period of 2 months, the results in all 3 cases were determined to be unappreciable and unsuccessful. The patients were subsequently treated with hypnoanalysis including ideomotor questioning, regression to onset, and reframing followed by direct suggestions under hypnosis (DSUH) for healing of all affected areas except a negotiated index finger on the nondominant hand. This technique proved an effective treatment that extinguished the flair-ups in 6 visits or less over a period of 2 months. These cases were followed at intervals of up to 1 year and no evidence of relapse found. PMID- 15754866 TI - Haiku: language, communication, and hypnosis. AB - This article will illustrate how effective hypnotic communication closely resembles the Haiku form. Working with the Haiku form is an effective and dynamic approach that encourages the therapists to keep their awareness sharpened and observation astute. Haiku is not just a type of poetry; it is a way of looking at the world with a heightened level of attentiveness. Crafting effective and evocative hypnotic suggestions requires that the therapist become immersed in the world of passion, images, sounds, sights, opposites, humor, creativity, and perceptive consciousness. Enhancing our skills of observation is an important aspect of the continuing experience of the hypnotherapist. The Haiku method can help us enhance our observation and utilize what we observe in developing evocative hypnotic suggestions that help the client access their internal representational systems to stimulate their healing response. A systematized method for learning to write Haiku is presented. PMID- 15754868 TI - Reflecting on tomorrow's nurses. PMID- 15754869 TI - Emergency ventilation of the tracheostomy patient, part II: a story of tracheotomy history and emergency airway management--advocating education for emergency resuscitation. AB - Over a period of 5000 years, dramatic changes have occurred in airway management, tracheotomy procedure terminology, indications, techniques, instruments, settings where procedures are performed, tube design and patient outcomes. Specialized knowledge and skills necessary to safely care for tracheostomy patients and to provide effective respiratory resuscitation are reviewed. The purpose of this paper is to document the history of the tracheotomy as a backdrop for understanding patient management. Recommendations for staff education regarding emergency ventilation of the tracheostomy patient are presented. PMID- 15754871 TI - Patient education. Allergy precautions in the home. PMID- 15754870 TI - "Code 99"--SOHN chapter extreme makeover. AB - Regional Chapters, both old and new, often experience a lack of participation and interest despite large membership numbers. Often, a few overworked members keep the Chapter afloat. This article, co-authored by two Chapter leaders, explores reasons for these changes and offers some tips for an "Extreme Chapter Makeover". PMID- 15754872 TI - Cortical mapping of EEG alpha power using a charge layer model. AB - This study aims to investigate how the blurred scalp alpha power distribution can be segregated on the cortex. EEG recorded from 11 subjects with their eyes closed were utilized to reconstruct the cortically equivalent Charge Layer (CL). Further, the power maps of scalp potential as well CL were generated by Fourier transform. The results showed a distinctly different CL power distribution from that of the scalp potential power, the cortical alpha electric activities were separated significantly into the left and right occipital regions, and in some cases activities at parietal regions were also clearly visibly discriminated. We concluded that the CL model is sensitive, and the blurred scalp activities can be further delineated into separate regions. PMID- 15754873 TI - Use of principal component analysis in the frequency domain for mapping electroencephalographic activities: comparison with phase-encoded Fourier spectral analysis. AB - Principal component analysis (PCA) can separate multichannel electroencephalographic (EEG) epochs into linearly independent (temporally and spatially noncorrelated) components. Results of PCA include component time-series waveforms and factors representing the contribution of each component to each electrode; these factors may be displayed as contour maps representing the topographic distribution of each component. However, PCA often does not achieve the most useful separation of components. PCA may be performed in the frequency domain to potentially improve results. After inspecting principal components of the frequency spectra, spectral values in a selected frequency range are multiplied by a chosen factor to emphasize (or de-emphasize) these frequencies and PCA is redone, promoting the separation of different frequencies into different components. Phase-encoded Fourier spectral analysis (PEFSA) uses multichannel complex Fourier spectra (amplitude and phase) to obtain positive or negative (phase-encoded) potentials at each electrode for any selected frequency. These may be displayed as a contour map representing the topographic distribution of the selected frequency. Applying both techniques, we found that EEG activities of differing frequency were readily separated by PEFSA, while standard PCA often mixed activities with different frequencies into a single component. However, frequency-domain PCA gave a component whose spatial distribution well matched PEFSA results. PCA is superior to PEFSA for separating activities with overlapping frequencies but differing spatial distributions. Preservation of phase information is an advantage of PEFSA and PCA over topographic maps that represent only amplitude (or power) at a given frequency. PCA or PEFSA maps can serve as a starting point for source localization. PMID- 15754874 TI - Scalp field potentials of human pain: spatial effects and temporal relation in finger stimulation. AB - In the present study, the spatial extent and temporo-spatial correlations of the human brain responses were investigated by electrically stimulating thumb (D1) and little finger (D5) under painful and non-painful intensity levels. High density (124-ch) somatosensory evoked potentials (SEPs) were recorded (-50 to +450 ms) from 15 healthy male volunteers. Early (0-50 ms), and late phases (150 450 ms) of the responses were analyzed. Peak stages 33 ms, 42 ms, 210 ms, 328 ms for D1; and 33 ms, 44 ms, 240 ms, 350 ms for D5 were isolated from a compressed waveform; the relationships within stages and phases were investigated using topography, Focal maximum Amplitude (FA, single central site at 0.2 cm2) and Area Magnitude (AM, summated FA and amplitudes of neighboring four sites in a region of nearly 9.9 cm2 area). In the early phase, the response of FA at 33 ms was significantly higher during painful than non-painful stimulation for both D1 and D5. In the late phase D1 (210 ms and 328 ms), and D5 (240 ms and 350 ms) the spatial areas of activation were significantly enlarged from non-painful to painful stimulation. For temporo-spatial relationship, D1 at 33 ms of early phase showed a significant correlation between the negative maximal site (sink) and the positive maximal site (source) under both non-painful and painful stimulations. For the late phase, the AM potentials at N2 correlated with that of P2 for both D1 and D5 under painful stimulation. The focal effects in FA of contralateral early potential indicates a shallow dipole in the primary somatosensory area of SI, while the large spatial extent in AM indicates a deep dipole of the putative cingulum activation under painful stimulation. No correlation between early and late activities implied that both activations are operated independently at the early SI and late cingulate processing of evoked pain. PMID- 15754875 TI - Localization of brainstem auditory evoked potentials in primates: a comparison of localization techniques applied to deep brain sources. AB - The objective of this study was to evaluate the performance of source localization techniques through localization of deep brain sources. To accomplish this, two replications of a brainstem auditory evoked potential (BAEP, left ear 60 dB nHL clicks) were recorded from five normal rhesus monkeys. We analyzed waves III and IV, as this portion of the BAEP corresponds to the deepest signal. Data were analyzed using five different source localization techniques: Moving Dipoles, Fixed Dipoles, MUSIC (Multiple Signal Classification) dipole scan, LORETA (Low Resolution Tomography), and LCMV (Linearly Constrained Minimum Variance) spatial filtering. The moving dipole, fixed dipole and MUSIC solutions were found to be, on average, 25.1 mm from the brainstem generators. LORETA detected sources within the brainstem 65% of the time. However, 90% of these localization results also included false detections defined as regions of the brain that were more than 2 cm away from the auditory pathway. LCMV included the brainstem in 90% of the trials and false detections in 40% of the cases. These findings indicate that evoked electrical activity from deep brain sources can be localized with cm accuracy. The dipole methods performed better than LORETA and LCMV. Given the depth and amplitude of the sources analyzed in this study, these results can be interpreted as an upper bound on the accuracy of each technique. PMID- 15754876 TI - Voltage-based versus factor score-based source localization analyses of electrophysiological brain activity: a comparison. AB - Though, traditionally, electrophysiological recordings have been limited to provide temporal information on neural activity, the development of mathematical algorithms capable of solving the inverse problem is facilitating, in recent years, the access to spatial information (i.e., on the origin of neural activation). This study explored a new strategy in order to increase the reliability of inverse problem solutions: applying these algorithms on factor scores (and not on voltages), a parameter that can be defined as "clean amplitude". Factor scores derive from Principal Component Analysis (PCA) applied to event-related potentials (ERPs). The main advantage of PCA is its capability to extract and quantify ERP components free of the influence of adjacent or subjacent components. The LORETA algorithm for source localization was applied on peak voltage, average voltage and factor scores for the motor potential recorded from 25 subjects, who had to repeatedly press a button with their right hand. The solutions given by LORETA in these three modalities were compared. The motor potential, a negative wave that begins just before any voluntary movement and is centrally distributed in the scalp, is particularly useful to the scope of this study, since its origin is known: contralateral motor cortex. Results show that the three modalities (peak voltage, mean voltage and factor scores) provided the same main focus (left motor cortex), though the "cleanest" solution (i.e., the main focus was more salient with respect to other secondary, noisy foci) was achieved by the factor score-based LORETA. PMID- 15754877 TI - Non linear analysis of magnetoencephalographic signals as a tool for assessing malignant lesions of the brain: first results. AB - BACKGROUND AND PURPOSE: Non linear signal analysis is a powerful technique that reveals qualitative and quantitative differentiations between different dynamical systems (biological or otherwise). Presented here are the first results of a work in progress to investigate the Magnetoencephalograms (MEG) from patients with malignant CNS lesions and from healthy volunteers. METHODS: We present MEG recordings of 10 patients diagnosed with malignant CNS lesions and the corresponding ones from 10 healthy volunteers. A 122-channel SQUID biomagnetometer in an electromagnetically shielded room was used to record the MEG signals and the Grassberger-Procaccia method for the estimation of the correlation dimension was applied on the phase space reconstruction of the recorded signal from each patient. RESULTS: Evidence linking MEG signal characteristics (existence of low dimensionality chaotic dynamics) with the existence of the tumour was found from this analysis. CONCLUSION: The obtained results substantiate our hypothesis of a relation between tumours of the brain and the mathematically chaotic nature of the neural dynamics derived from their MEG recordings. PMID- 15754878 TI - An anti-Freudian approach to improve life for dementia patients. PMID- 15754879 TI - Medicare prescription Rx cards: making difficult choices. PMID- 15754881 TI - Revolution in neurosurgery marches into refractory psychiatric illness. PMID- 15754880 TI - The move to screen: underused or heavyhanded? PMID- 15754882 TI - How is case management. Working? Ask consumers directly. PMID- 15754883 TI - Workplace presenteeism: How behavioral professionals can make a difference. PMID- 15754884 TI - Targeted training the basis for NCCBH Conference's success. PMID- 15754885 TI - Implementing CQI at the healthcare provider level. PMID- 15754886 TI - A learning community for women in recovery. PMID- 15754887 TI - Oral dyskinesia: a clinical overview. AB - PURPOSE: Dentists may be the first health care professionals to recognize unusual and abnormal oral movements collectively termed oral dyskinesias. The aims of this clinical overview are to raise the dental community's awareness about this important and complex topic and describe the clinical features and management of the main entities. MATERIALS AND METHODS: A MEDLINE search of the different entities reported in the English and French literature was conducted. The main findings of a field study on oral dyskinesia were also reviewed. RESULTS: Involuntary movement disorders are often drug related. In other cases, excessive oral movements may occur at any age in relation to various neuropsychiatric conditions. Orofacial dystonia apparently triggered by dental procedures has also been reported. Edentulousness has been associated with oral stereotypes. In a survey of 352 edentulous elderly individuals attending daycare centers, only 7% displayed visible oral sterotypes, and ill-fitting dentures were suggested as a possible triggering factor for the majority. CONCLUSION: A multidisciplinary evaluation is desirable in the care of individuals with oral dyskinesia and in the selection of those who may benefit from a prosthodontic approach. A good knowledge of potentially offending drugs may allow avoidance of unnecessary procedures. PMID- 15754888 TI - An economic evaluation of implant treatment in edentulous patients-preliminary results. AB - PURPOSE: Edentulous patients with denture problems benefit from implant treatment with overdenture prostheses. The aim of this prospective study was to investigate a method of analyzing cost effectiveness in dentistry. As an example, overdenture treatment with two or four implants was compared to the conventional complete denture (CD). MATERIALS AND METHODS: In a self-selected trial, 20 patients each were treated with implant-retained overdentures (two implants, IRET), implant supported overdentures (four implants, ISUP), or CDs (control group) in the edentulous mandible. A cost-effectiveness analysis was performed from the patient's perspective, with a time horizon of 6 months. Direct health-care costs were calculated in Swiss Francs (in 2000), and effects were defined as improvements in perceived chewing ability compared with the baseline value before treatment (measured on a VAS). Point estimates for mean incremental cost effectiveness ratios were complemented with cost-effectiveness acceptability curves to account for uncertainties associated with costs and effects. RESULTS: Mean incremental costs were CHF 4,329 (IRET-CD), CHF 13,360 (ISUP-CD), and CHF 9,031 (ISUP-IRET); these cost differences were all statistically significant. The mean incremental effects at 6 months were 19% (IRET-CD), 23% (ISUP-CD), and 4% (ISUP-IRET). Incremental cost-effectiveness ratios were CHF 228 (IRET-CD), CHF 581 (ISUP-CD), and CHF 2,258 (IRET-ISUP) per percentage increase in chewing ability. CONCLUSION: From an economic point of view, IRETs were more attractive than ISUPs. The latter were associated with a statistically significant improvement in perceived chewing ability compared to CDs, but at substantially higher costs. PMID- 15754889 TI - Effect of water storage on the flexural properties of three glass fiber reinforced composites. AB - PURPOSE: The effect of water sorption on the flexural strength and flexural modulus of three fiber-reinforced composites was studied. MATERIALS AND METHODS: Bar-shaped specimens of each material were prepared according to the manufacturers' instructions. The flexural strength and modulus of each specimen were tested after the specimens were immersed in distilled water for 1, 7, 60, and 180 days. SEMs were taken to examine the mode of failure. The volume percentage of fiber content of each fiber-reinforced composite was experimentally estimated. RESULTS: The ascending order of flexural strength and modulus among the materials was generally: FibreKor < Stick < Vectris. The flexural strengths of Stick and FibreKor specimens at 1 and 180 days were not significantly different. Although the 180-day Vectris specimens possessed significantly lower flexural strength than the 1-day specimens, the flexural strengths of Vectris specimens at 1, 7, 60, and 180 days remained significantly higher than those of Stick and FibreKor. The difference in flexural modulus for each fiber-reinforced composite related to the duration of water immersion was not significant. CONCLUSION: Water immersion affected the flexural strengths of the three composites to a different degree but did not affect their flexural moduli significantly. For each duration of water immersion, the flexural property of the fiber-reinforced composite, in ascending order of significant difference, was: FibreKor < Stick < Vectris. PMID- 15754890 TI - 5-year follow-up of a prospective clinical study on various types of core restorations. AB - PURPOSE: This study tested whether: (1) the survival rate of cast post-and-core restorations is better than the survival of direct post-and-core restorations and post-free all-composite cores; and (2)the survival of these buildup restorations is influenced by the remaining dentin height after preparation. MATERIALS AND METHODS: In a clinical trial, 18 operators made 319 core restorations in 249 patients. The restorations involved were: (1) cast post-and-core restorations; (2) direct post and composite core restorations; and (3) post-free all-composite cores. All restorations were made under single porcelain-fused-to-metal crowns. Treatments were allocated after dentin height assessment using balanced drawing. Failures were registered during a 5-year period. RESULTS: Fifteen restorations failed during the follow-up period. Five failures occurred during the first month; they were considered to be independent from clinical aging and excluded from further survival assessments. The overall survival was 96%+/-2%. No difference was found between the survivals of the different types of restorations. The factor "remaining dentin height" appeared to have a significant effect on the survival of post-and-core restorations (98%+/-2% survival for "substantial dentin height" vs 93%+/-3% for "minimal dentin height"). CONCLUSION: The type of post and core was not relevant with respect to survival. The amount of remaining dentin height after preparation influenced the longevity of a post and-core restoration. PMID- 15754891 TI - A 5-year prospective clinical study on core restorations without covering crowns. PMID- 15754892 TI - Prosthetic intervention in the era of microvascular reconstruction of the mandible--a retrospective analysis of functional outcome. AB - PURPOSE: The purpose of this retrospective study was to compare the functional outcomes of patients who had mandibular resection and reconstruction with and without prosthetic intervention, and to identify predictive factors that may have an impact on functional outcomes. MATERIALS AND METHODS: Two hundred twenty head and neck cancer patients who had undergone mandibular resection and reconstruction with at least 6 months of postoperative convalescence formed the basis of this retrospective review. Patients who did not receive prosthetic intervention formed group I (n = 142); those who received prosthetic intervention formed group II (n = 78). Functional outcomes were measured using four individual assessments (nutritional status, swallowing, masticatory performance, and speech) and one that combined the information from these assessments, the global measure of functional outcome (GMFO). Statistical analyses were used to compare the baseline characteristics and functional outcome between groups I and II and to analyze independent predictors for GMFO. RESULTS: Of the 220 patients reviewed, 78 (35%) had prosthetic intervention; group II patients had better individual functional outcome measures and GMFO. Use of a prosthesis remained associated with GMFO after controlling for other significant predictors; other independent predictors were xerostomia, number of remaining mandibular teeth, number of tooth to-tooth contacts, type of reconstruction, flap interference, and tongue defect. Patients who had fewer mandibular teeth and received a smaller prosthesis had better overall outcome than patients who received a larger prosthesis. CONCLUSION: Patients who had prosthetic intervention after mandibular reconstruction had significantly better functional outcomes than patients who did not receive prosthetic intervention, even after adjusting for confounding variables. PMID- 15754893 TI - Candida-related denture stomatitis: a pilot study of the efficacy of an amorolfine antifungal varnish. AB - PURPOSE: Candida albicans is a component of the normal oral microflora, but local and systemic factors can transform this commensal C albicans to a pathogen. The most frequent cause of Candida opportunistic infections (candidiasis) is dentures, especially if poorly fitting or poorly cleaned. Management of oral candidiasis depends on an accurate diagnosis, identification and elimination of predisposing factors, and, often, use of antifungal agents. The aim of this study was to examine fingernail varnish, currently used for onychomycosis therapy, to reduce the fungal colonization in prosthetic biofilms. MATERIALS AND METHODS: A varnish containing 5% amorolfine was applied once or twice a week for 6 months in six patients affected by nystatin-resistant denture-related stomatitis. In all six patients, the prostheses had previously been removed at night, and daily antimycotic topical therapy with nystatin had failed to resolve the stomatitis; after 30 days, these patients all showed persistence of candidal stomatitis. RESULTS: After 1 month, five of the six patients were negative for Candida; this situation was unchanged in the following monthly controls. Only in the patient with suspected Sjogren syndrome was oral Candida found 15 days after the last varnish application. None of the patients had any complaints about the medication. CONCLUSION: This varnish containing 5% amorolfine, applied once or twice a week for 6 months, was able to suppress the nystatin-resistant denture related stomatitis. PMID- 15754894 TI - Effect of surface treatments on the resin bond to zirconium-based ceramic. AB - PURPOSE: This study tested the hypothesis that the tribochemical silica coating on ceramic surfaces increases the bond strength of resin cement to a glass infiltrated zirconium-based ceramic. MATERIALS AND METHODS: Fifteen blocks of In Ceram Zirconia from CEREC InLab (5 per group) and 15 composite blocks (Z-250) 5 mm x 5 mm x 4 mm were made. The ceramic surfaces were polished, and the blocks were divided into three groups: (1) airborne abrasion with 110-microm aluminum oxide particles; (2) Rocatec system, tribochemical silica coating; and (3) CoJet system, tribochemical silica coating. The ceramic blocks were cemented to the composite blocks using Panavia F according to the manufacturer's specifications. All samples were stored in 37 degrees C distilled water for 7 days and later sectioned in two axes using a diamond disk under cooling to obtain specimens with a cross-sectional area of approximately 1 mm2 (n = 45). Each specimen was then attached with cyanoacrylate glue to an adapted device for the microtensile test, which was carried out on a universal testing machine. RESULTS: The results were subjected to ANOVA and Tukey's test. Group 2 (23.0+/-6.7 MPa) and group 3 (26.8+/ 7.4 MPa) showed greater bond strength than group 1 (15.1+/-5.3 MPa). There was no significant difference between groups 2 and 3. All failures were in the adhesive zone. CONCLUSION: The hypothesis was confirmed--the tribochemical systems increased the bond strength between Panavia F and In-Ceram Zirconia. PMID- 15754895 TI - The oral pigmentation chart: a clinical adjunct for oral pigmentation in removable prostheses. AB - PURPOSE: Non-Caucasian patients exhibit different characteristics of oral pigmentation and may request that the acrylic resin parts of their dentures look natural, simulating the original mucosal color. Tooth loss, bone resorption, and lack of attached gingiva may, however, make it difficult to determine what the original pigmentation was like. The purpose of this investigation was to study the distribution in oral pigmentation around the natural dentition in non Caucasians, in a preliminary effort to classify these variations into a chart of oral pigmentation, and to analyze its reproducibility. MATERIALS AND METHODS: For the study, 106 dentate non-Caucasians were selected from two universities: ACTA (patient group) and UCLA (nonpatient group). A pigmentation scheme was devised on the basis of half of the participants, and the others were divided into categories by four observers independently. Cohen's kappa was then calculated. RESULTS: On the basis of information obtained from the ACTA participants, six categories of mucosal pigmentation were defined. The kappa statistics for the four observers varied from .58 to .79 for intraobserver agreement and from .15 to .55 for interobserver agreement. CONCLUSION: The Oral Pigmentation Chart is a simple device that makes it possible to simulate oral pigmentation in the acrylic resin parts of removable dentures. The reproducibility appeared to be acceptable when clinician and dental technician were calibrated. Patients can be offered a choice of the kind of pigmentation geography they want in their removable prostheses. PMID- 15754896 TI - Bond strengths of veneering ceramics to reinforced ceramic core materials. PMID- 15754897 TI - Comparison of the color of ceramics as measured by different spectrophotometers and colorimeters. PMID- 15754898 TI - Prosthodontics and the patient: what is oral rehabilitation need? Conceptual analysis of need and demand for prosthodontic treatment. Part 1: a conceptual analysis. AB - PURPOSE: The concepts of need and demand are central in studies on dental care. In the literature, a normative definition is often used, but it pays little attention to the individual's personal comfort and quality of life. Need and demand for prosthodontic services are difficult to measure, as prosthodontic treatment is highly individual and not closely related to edentulousness. Need, however defined, does not always lead to demand for treatment, depending on a variety of factors. MATERIALS AND METHODS: The present article is part of a larger study in which the intention is to evaluate need and demand for prosthodontic treatment among the participants in a 1989 and 1999 longitudinal study of a population sample. As the first step, this article reports a conceptual analysis of the need concept from the literature. RESULTS: Need is stated as socially established in the interaction between patient and clinician. It makes demand dependent on available treatment options from the care provider and society. In the prosthetic treatment decision-making process, the emancipatory perspective with the patient-clinician dialogue is of utmost importance to achieve an optimal treatment result. CONCLUSION: The professional attitude toward need must be that there is no true objective or subjective need. Need is established only in a communicative dialogue with mutual respect between the professional and the patient. PMID- 15754899 TI - 'Code' called contrary to pt.'s advance directives. PMID- 15754900 TI - Why wasn't 'continuous treatment' doctrine applied? Case on point: Knox v. Greenville Hospital System, 2005 WL 20007 S.E.2d--SC. PMID- 15754901 TI - PA: Was LPN fired or did she quit?: LPN quit!--unemployment benefits denied! PMID- 15754902 TI - The nurse's role in determining testamentary capacity. Case on point: In re. purported last will of Patton, 2004 WL 3030543 N.E.2d--DE. PMID- 15754904 TI - [Racing?]. PMID- 15754903 TI - [Equine exudative canker: an (auto-)immune disease?]. AB - Equine canker is a chronic, hyperplastic, exudative pododermatitis affecting one or more feet. Although many causes and treatments have been suggested, the cause of the disease is still unknown and most probably multifactorial. Local treatments include radical surgical debridement of the diseased hoof tissue and application of caustic substances, antibiotics, and pressure bandaging. Nevertheless, the number of recurrences is high (45%). This article presents a 3 year-old New Forest pony-cross mare in which all horny structures (frogs, coronets, spurs, chestnuts) of all feet were affected. Bacteriological and fungal cultures of the frogs were found negative for the pathogens tested. Papilloma virus was not found. Clinical findings raised the hypothesis that the non specific hyperplastic inflammation of these horn-like structures might have been caused by an (auto-)immune reaction. On the basis of the clinical findings, the pony was treated with surgical debridement of the frogs of a diagonal pair of feet and oral administration of prednisolone (1 mg/kg sid). The frogs, coronets, spurs, and chestnuts of all four feet healed completely within 8 weeks, thus making an (auto-)immune reaction more likely. In conclusion, this case report raised the hypothesis that an aspecific, hyperplastic inflammation of all four feet ('equine canker') and other horny structures may be caused by an (auto )immune reaction, and that corticosteroids (prednisolone 1 mg/kg sid per os) are effective as treatment. PMID- 15754905 TI - [Rabies in bats]. PMID- 15754906 TI - [Honey in treatment of skin wounds]. PMID- 15754907 TI - [Interview with Tony Achterkamp, chairman of Platform for Responsible Animal Husbandry. Text by Paul Overgaauw]. PMID- 15754908 TI - [Symposium 'Health care in biological animal husbandry']. PMID- 15754909 TI - [New limb sparing therapy for osteosarcoma in the dog]. PMID- 15754910 TI - [Comfortable conversation]. PMID- 15754911 TI - "Reverse the curse". PMID- 15754912 TI - Nurse manager survival in an age of new health care priorities. PMID- 15754913 TI - Pesticide drift. PMID- 15754914 TI - Factors influencing pain, physical function and social functioning in patients with osteoarthritis in southern Italy. AB - The aim of this study was to assess factors influencing bodily pain (BP), physical function (PF) and social functioning (SF) in patients with osteoarthritis (OA) from southern Italy A total of 1,782 patients (mean age 66.08 years, 570 men and 1,212 women) with knee, hip, spine or hand OA underwent a structured assessment comprising demographic data and the Short Form 36 (SF-36) BP, PF and SF scales. Separate multiple linear regression models were employed for statistical analysis. The mean disease duration was 9.18 years and the mean body mass index (BMI) was 27.06. The mean BP, PF and SF scores of 34.93 (SD 19.37), 63.58 (SD 26.53) and 47.89 (SD 21.83) for the study subjects were substantially lower than those expected for the general Italian population. Subjects who were younger with a shorter disease duration and lower BMI had better PF and SF Younger subjects with a lower BMI and a longer disease duration had less BP. Female sex was associated with more BP, worse SF and better PF. In conclusion, demographic and disease-related factors influence BP, PF and SF in southern Italian patients with OA. PMID- 15754915 TI - Comparison of new different assay systems for thyrotropin receptor antibodies with reference to thyroid-stimulating antibodies and thyroid stimulation-blocking antibodies in Graves' disease. AB - The aim of our study was to evaluate the diagnostic sensitivity of the new thyrotropin receptor antibody (TRAb) assays (Cosmic TRAb CT, ELISA and Yamasa DYNOtest TRAb). TRAb was positive in 43 of 44 (97.7%) untreated patients with Graves' disease by both TRAb CT and/or ELISA and NYNOtest TRAb. Thus the new TRAb assays were clearly more sensitive than the conventional assay (positivity: 85%). There was a strong positive correlation between the data obtained in TRAb CT and/or ELISA and those obtained in DYNOtest TRAb (r = 0.942, p < 0.0001). There was a significant correlation between the new TRAb and TSAb (r = 0.696, p < 0.0001). Although there was a significant correlation between the new TRAb and thyroid stimulation-blocking antibody (TSBAb), the correlation coefficient was low (r = 0.605, p < 0.0001). The increased sensitivity of the new TRAb assays for Graves' disease provides an advantage over conventional assay. PMID- 15754916 TI - Life-threatening renal failure caused by vasomotor nephropathy associated with nonsteroidal anti-inflammatory drugs. AB - The purpose of this study was to evaluate the prevalence of life-threatening renal failure (RF) caused by vasomotor nephropathy associated with nonsteroidal anti-inflammatory drug (NSAID) treatment (NSAID-RF) and risk factors for this renal impairment in an inception cohort of patients with recently diagnosed uremia treated by emergency hemodialysis in a prospective regional study. There are few published data on this phenomenon. Two hundred fifty-six patients (137 men, 119 women, mean age 68 years [22-95 years]) with acute uremia were treated with emergency hemodialysis in the intensive care unit over a period of 70 months. The patients were from an area of 231,000 inhabitants. Of the 256 patients, clinical data from a group of 79 patients with medical-type renal failure were analyzed in detail. The prevalence of NSAID-RF was 8%. This prevalence decreased to 4% when patients without any other medication affecting compensatory renal hemodynamics were considered. Moreover when nonpharmacological insults were not taken into account the prevalence decreased to only 1.6%. In 80% of the patients with NSAID-RF, nonpharmacological insults contributed to renal impairment. Both hypotension of cardiac etiology and dehydration/hypovolemia were present in 25% of the patients with this type of RF while urinary tract obstruction was seen in 1%. In 75% patients with NSAID-RF the underlying nephropathies were identified. NSAID-RF was not frequent. The population at greatest risk for renal functional alteration associated with NSAID therapy included patients with dehydration/hypovolemia, hypotension of cardiac etiology and those with pre-existing renal impairment, especially with vascular and analgesic nephropathy. PMID- 15754917 TI - Working women are better responders to beta-blocker monotherapy of mild hypertension than men. AB - The aim of this single-blind study was to compare the efficacy of betaxolol treatment (20 mg/day) on 24-h blood pressure profiles in working men and women with mild hypertension (grade 1 acc. ESH/ESC/JNC 2003), A group of 11 men and 11 women with a mean age 47+/-5 years underwent 24-h blood pressure monitoring after 8 days of placebo and after 20 days of treatment. A significant reduction (p < 0.05) in blood pressure was found for 11 h in men and 15 h in women (systolic) and 9 h in men and 13 h in women (diastolic). There was a tendency for a greater mean reduction in women (9.6/8.0 mmHg in men versus 12.9/7.4 mmHg in women). Diastolic blood pressure variability was significantly reduced in women (9.9 versus 13.1, respectively, p < 0.002) with a tendency for systolic blood pressure variability reduction (13.0 versus 15.1). The smoothness index for systolic blood pressure was higher in women (1.0/0.74 versus 0.64/0.61). A better response for betaxolol treatment 20 mg/day was observed in women in terms of target organ damage: a longer period of significant blood pressure reduction, lower variability and a tendency toward a greater reduction. PMID- 15754918 TI - Molecular mechanism for accelerated atherosclerosis in diabetes and its potential therapeutic intervention. AB - Diabetes is associated with a marked increase in the risk of atherosclerotic vascular disorders, including coronary, cerebrovascular and peripheral artery disease. Macrovascular complications could account for disabilities and high mortality rates in patients with diabetes. There is a growing body of evidence to indicate that tight blood glucose control has no more than a marginal impact on cardiovascular disease in diabetes. In this review, we discuss the molecular mechanisms for accelerated atherosclerosis in diabetes and the potential therapeutic interventions, especially focusing on postprandial hyperglycemia and advanced glycation end products (AGEs). PMID- 15754919 TI - A mixed picture. PMID- 15754920 TI - Basic instincts. PMID- 15754921 TI - Good counsel. PMID- 15754922 TI - In the dark. PMID- 15754923 TI - Missing pieces. PMID- 15754924 TI - Communication about sexual problems in male patients with multiple sclerosis. AB - AIM: To explore the reasons given by a disability, assessment and rehabilitation team (DART) for perceived difficulties in discussing sexual problems with male patients with multiple sclerosis (MS), and to compare them with the views of a sample of patients. METHOD: Data were gathered from a multidisciplinary team who were responsible for assessment and rehabilitation of patients with MS, using a focus group and semi-structured interviews with 11 male patients. Data were analysed thematically, with reference to Annon's (1976) model of communication. FINDINGS: Patients felt a need to discuss problems with sexual dysfunction, and would welcome the introduction of the topic in a straightforward way. Health professionals were worried about intrusiveness and lack of information and training. Team members and the patients indicated that nurses are the best people to deal with these issues. CONCLUSION: Rehabilitation teams need training to deal with patients' sexual problems, and the nurse is likely to be identified as the specialist in this area. PMID- 15754925 TI - The district nurse's role in managing patients with heart failure. AB - Heart failure is a complex disorder that affects patients and their carers in many ways. Recommendations for improving the quality of care for people with heart failure are set out in the government's National Service Framework for Coronary Heart Disease (Department of Health (DH) 2000a) and in the guideline commissioned by the National Institute for Clinical Excellence (NICE 2003a), which describes best practice for the management of patients with heart failure. This article discusses the role of the district nurse in meeting the challenges of the NICE guideline, and suggests that district nurses are better placed than other health professionals in primary care to have a central role in delivering such care. The contribution of district nurses to the management of patients with chronic diseases and those who are terminally ill at home is also discussed. The impact of The NHS Cancer Plan (DH 2000b) on the care of cancer patients in primary care is examined briefly to ascertain whether the NICE guideline is likely to have a similar impact on the scope and capacity of district nursing. PMID- 15754926 TI - Headache. AB - This article examines the different types of headache that patients may experience, distinguishing between common types of headache and those that are life-threatening. PMID- 15754927 TI - Wound infection and colonisation. AB - Many wounds seen by nurses will involve infection and colonisation. To enable nurses to correly assess and manage these wounds, infection and colonisation are explained and options for management discussd. PMID- 15754928 TI - A guide to emollient therapy. AB - Although the benefits of emollient therapy have been widely acknowledged in dermatology treatments (Cork 1997), emollients have tended to be undervalued and under-used in general care and wound management. Healthy skin is an essential part of patient wellbeing, and emollient therapy is integral to the management of skin problems. The integration of tissue viability and dermatology services in the Plymouth Hospitals NHS Trust provided an opportunity to evaluate how and why emollient therapy is implemented. In practice, the authors have identified that healthcare practitioners need guidance and advice on emollient therapy to improve their care delivery. This article is based on the emollient therapy currently used in the authors' trust. PMID- 15754929 TI - Time to change. PMID- 15754930 TI - Life giver. Interview by Brian McCalden. PMID- 15754931 TI - Looking towards a changing future. PMID- 15754933 TI - Should nurses be role models for health? PMID- 15754934 TI - The CPD challenge. PMID- 15754935 TI - Treating complex wounds at home. PMID- 15754936 TI - 'These awards bring national recognition'. PMID- 15754938 TI - Wound infection. PMID- 15754937 TI - Raising awareness of Raynaud's phenomenon and scleroderma. AB - The 2005 Raynaud's and scleroderma awareness campaign aims to highlight Raynaud's phenomenon, a common episodic circulatory disorder in which the small blood vessels in the extremities are oversensitive to changes in temperature. For a small number of people, Raynaud's is one of the earliest symptoms of scleroderma, so people with Raynaud's require prompt assessment and diagnosis. PMID- 15754939 TI - Developing a chest pain team to fast track AMI patients. AB - Rapid identification and treatment of patients found to have acute myocardial infarction (AMI) saves lives. This article identifies some of the clinical leadership challenges in the development of a chest pain team to fast track AMI patients. PMID- 15754940 TI - Tracheal suction. PMID- 15754941 TI - Nystatin. PMID- 15754942 TI - Managing acute and chronic pain in sickle cell disease. AB - Acute painful episodes among patients with sickle cell disease may occur in any body part or several sites simultaneously. The reasons for inadequate pain relief for this patient group are complex. Prompt, accurate pain assessment is the cornerstone of effective pain management and a comprehensive, multidisciplinary team strategy is essential, particularly for managing patients with frequent acute pain or chronic pain. PMID- 15754943 TI - The importance of thorough physiological assessment. PMID- 15754944 TI - The effectiveness of the national cleanyourhands campaign. AB - The National Patient Safety Agency (NPSA) is a key contributor to the national drive to reduce avoidable health care-associated infections (HAIs) through the implementation of the cleanyourhands campaign. The campaign is based on international evidence of the most effective methods for achieving a sustained year-on-year improvement In compliance with hand hygiene (Larson et al, 2000; Pittet et al, 2000) and is targeted at NHS staff working in the acute sector. PMID- 15754946 TI - Best practice in urinary catheterisation and catheter care. AB - In May 2002 a series of 'best practice statements' to encourage a consistent and cohesive approach to care was launched by the former Nursing and Midwifery Practice Development Unit (NMPDU). PMID- 15754945 TI - Minimising cross-infection risks associated with beds and mattresses. AB - The spread of infection can be associated with inanimate objects in the environment, referred to as fomites. These include beds, mattresses, curtains, bedpans and sphygmomanometers, all of which may be contaminated with micro organisms (Wilson, 2001). PMID- 15754948 TI - Making the most of a careers event. PMID- 15754947 TI - The special needs of children receiving intravenous therapy. AB - Although the principles of preventing infection associated with intravenous therapy are the same for all patient groups, there are some notable differences when caring for babies and infants. The incidence of nosocomial infections in children is proportional to their age, with the highest incidence occurring in neonates and children under one year of age. PMID- 15754949 TI - [The differences and regularities of life-span dynamics for human and animal populations]. AB - The dynamics of mortality of several generations of man, Wistar rats and line Canton-S fruit flies Drosophila melanogaster was investigated. The result was, that the correlation between sample mean and sample variance was negative for man, positive for flies and was absent for rats. To fit the survival curve we chose two-parametric Gompertz function. Starting from Strehler-Mildwan correlation (linear correlation between Gompertz function parameters), we obtained theoretical dependence between life-span mean and variance. This dependence had complex shape and points, corresponding to different species were situated at different segments of this curve that is corresponding to the various sign of correlation between these characteristics. Increasing of the mean man life-span attribute usually to advances in medicine and social environment. Associated with this the decreasing of the life-span variance, as it follows from this paper, depends on parameter point location on parametric plane, i.e. from biological feature of species. PMID- 15754950 TI - [Are Nobel prize winners gettting older? Mathematical analysis of age and life span of the Nobel prize winners, 1901-2003]. AB - Data on the distribution by the age of 647 Nobel prizewinner in physics, chemistry, physiology or medicine, economics and literature and time dynamics of the age during 1901-2003 are presented in the paper. The mean survival and the expected life span of the prizewinners was also calculated. The mean life span of the Nobelists in natural sciences steadily increases from the first to the fourth quartile of the 20th century. The rate of the prizewinners selected at the age of 61 and more years increases from the 23% at 1901-1925 to 53.1% at 1975-2003, whereas the rate of the winners selected before the age of 40 years decreases from 19% to 2.7% during the same period. Analysis of available data on the Nobel Prizes for chemistry or physiology or medicine has shown that the majority of the Prizes was awarded to researcher discovered his main results at the age of 31-40 (41.8% and 47.4%, correspondingly). The mean interval between a discovery and a selection for Nobel Prize in chemistry increases from 12.6 years in 1901-1925 to 23.7 years in 1975-2003, whereas corresponding parameters for prizewinners in physiology or medicine increases from to 16.9 to 20.1 years. The mean life span of deceased Nobelists is 77.8 years, varying from 76.0 years in chemists to 84.8 years in economists. The mean life span after the selection is 20.3 years (from 13.6 years in writers to 24.6 in physicists). The expected life span after the selection adjusted by the age and the year of the selection of the Nobelists in physics, chemistry, economics and literature in 1959-1999 failed to reveal any increase, whereas the life expectancy of the prizewinners in physiology or medicine is increased as compared with the male population of USA. PMID- 15754951 TI - [Historical dynamics of age-related mortality: 1. Evolution of the specific lifespan of the Finnish population]. AB - Historical dynamics of parameters of age-related mortality of Finnish men and women for the period 1952 to 2000 is studied. The Gompertz equation is found to describe age-related mortality of men quite exactly during the historical interval 1952 to 1989. From 1990 to 2000 for men and during the entire interval for women the Gompertz-Makeham equation is more exact. The constant component of mortality is found to be increasing since 1990 for men and since 1980 for women. The presence of the Strehler-Mildvan correlation is shown. There are 3 evidently separated periods: 1) from 1952 to 1971, 2) from 1972 to 1980, 3) from 1981 to 2000. The correlation has the highest value in the 1st and 3rd periods. During the 2nd, transitional, period the correlation value is minimal. An opinion is expressed that it's not correct to treat the incline coefficient of correlation dependence as a specific lifespan since it takes on different values even for a single country, though the values are stable during certain historical periods. PMID- 15754952 TI - [Analysis of the life span distribution mode in 128 successive generations of D. melanogaster]. AB - A large body of experimental data consisting of 116 samples (sets) of individual life span (LS) values of D. melanogaster from the same laboratory strain Canton-S was analyzed. In total, 10180 Drosophila flies (5100 females and 5080 males) were studied. Each of 58 pairs of samples belonged to a definite generation in a continuous succession where every next generation was an offspring of the preceding one. Mathematical simulation made it possible to demonstrate that both the normal (Gaussian) and Gompertz distributions were equally good approximations of the experimental data. Both of them adequately described the LS distributions in laboratory populations of D. melanogaster. The confidence intervals for absolute deviations of the theoretical distributions from experimental ones were small (4-5%). In other words, the approximation error was no more than 5% in either case. The estimation of the dependence of approximation quality on the LS in the original (experimental) population showed that the normal distribution was preferable, because, in this case the absolute deviation from the experimental distribution was independent of the LS in the original population. PMID- 15754953 TI - [Numerical modeling of ideal cohorts of aging organisms obeying the Gompertz Makeham law in association with the Strehler-Mildwan correlation]. AB - The Gompertz-Makeham law (-dn/dt x l/n(t)=C+lambdae(gammat)) so as other genuine laws of Nature is strictly applicable only to ideal objects (populations and cohorts) analogously to laws of mechanics or thermodynamics, which are exactly true only for such physical abstractions as mass points or ideal gases. Therefore, a biologically meaningful interpretation of the parameters of this law is likely to be more important for understanding the aging process than devising of alternative analytical descriptions of biodemographic processes for the sake of a better fit only. Numerical modeling of ideal cohorts of aging organisms obeying the Gompertz-Makeha law makes it possible to differentiate possible real and apparent changes in mortality patterns that occur in human history and in evolution and are observed in gerontological experiments and to demonstratively show such effects as the dependency of longevity upon population size, the evolutionarily important possibility of reciprocal changes in the mean and maximal longevity, or detection of apparent changes in negatively correlated aging rate and vitality when the Makeham term is ignored, which is usual in demography. The basic difference between the Makeham term Cand Gompertz term lambdae(gammat) is suggested to be not that the former is constant, whereas the latter is age-dependent, but that the former comprises the contributions of inherently irresistible stresses to mortality, whereas the latter comprises the contributions of resistible stresses to mortality and shows how changes in the ability to resist them is translated into changes in mortality. PMID- 15754954 TI - [Neuroendocrinology of the stomach and aging]. AB - The survey contains information about basic stomach endocrine cells in normal condition and their changes in chronic atrophic gastritis and ageing. The data of modern research devoted to endocrine cells age-related changes are represented here and analyzed. The reasons that influence on endocrine cells age-related changes and the ways of their possible correction are described. PMID- 15754955 TI - [Skin photo-aging--molecular markers of epidermis and dermis]. AB - The review has deal with available data on the range of molecular markers of chronic photic damage and photo-ageing of the epidermis and dermis. PMID- 15754956 TI - [Circadian rhythms of pineal gland melatonin-forming function in healthy elderly people]. AB - Circadian rhythms of pineal gland melatonin-forming function were studied in healthy elderly people (n=31, age range 60-74 years). The blood plasma melatonin concentration was determined by means of radioimmune method at 3.00, 9.00, 15.00 and 21.00. The study revealed two types of circadian rhythms of plasma melatonin concentration: a high-amplitude (29%) and a low-amplitude (71%) ones. The high amplitude rhythm of melatonin-forming pineal gland function was observed in elderly people with a higher physical working ability and a lower functional age of cardiovascular system. It is advisable to conduct the correction of pineal gland melatonin-forming function in elderly people with low-amplitude circadian rhythm of plasma melatonin concentration. PMID- 15754957 TI - [Study of stability of lactate dehydrogenase isoenzymes in leucocytes of elderly people]. AB - In this paper was investigated stability of isoenzymes of lactatdehydrogenase (LDH) of leukocytes of peripheral blood of persons at the age of 20 years (n=10) and more than 65 years (n=8). Extraction of the leukocytes was conducted by centrifugation of human blood on a double gradient of density. Leukocytes were destroyed by hypotonic shock in combination with vibration. Separation of LDH isoenzymes from destroyed leukocytes was conducted by the method of native electrophores. The revealing of activity of LDH isoenzymes was conducted by the method of restoration of nitroblue tetrasolium. Isoenzymes stability was evaluated according to alteration of activity after the incubation of destroyed leukocytes with urea in various concentrations (1M, 2M and 3M). Was shown the presence of three isoenzymes of LDG (LDH-1, LDH-2, LDH-3) in human leukocytes. As the age-dependent alteration of activity of isoenzymes of lactatdehydrogenase in human leukocytes was shown. It can be caused by age-related changes of physico chemical properties of leukocytes' membranes and/or isoenzymes stability. Stability of LDH-3 was decreased with the age. Observed decrease of LDH-3 stability can be caused by age-related modification of M-subunits of the studied isoenzymes. PMID- 15754958 TI - [Effect of antidiabetic drug diabenol on parameters of biological age, life span and tumor development in NMRI and HER-2/neu mice]. AB - The effect of new antidiabetic drug Diabenol [9-diethylaminoethyl-2,3 dihydroimidazo-(1,2alpha) benzimidazol dihydrochloride] on life span and spontaneous tumor incidence in NMRI and transgenic HER-2/neu mice was studied. Female NMRI and transgenic HER-2/neu mice were given diabenol with drinking water (0.1 mg/l=approx. 10 mg/kg of b.w.) 5 times a week since the age of 2 months until natural death. The treatment with the drug failed influence body weight gain dynamics, food and water consumption and the body temperature in NMRI mice. Diabenol treatment slowed down age-related disturbances in estrous function and increased life span of all and 10% most long-living NMRI mice. The treatment with diabenol inhibited spontaneous tumor incidence (mammary gland and lymphomas mainly) and increased the mammary tumor latency. Dibenol treatment slowed down age-related changes in estrous function in HER-2/neu mice, failed influence survival of these mice and slightly inhibited the incidence and decreased the size of mammary adenocarcinoma metastases into the lung. Thus, long-term treatment with diabenol is safe and non-toxic in mice. The drug increases survival and inhibits spontaneous carcinogenesis in mice PMID- 15754959 TI - [Effect of selenium on reactions of elderly rats affected by low doses of radiochemical exposures]. AB - The 18-months-old female rats received with drinking water Hg2(NO3) in concentration of 1 mkg/l in account to metal and a preparation of selenium "Neoselen" (Na2SeO2) in concentration of 1.3 ml/kg/day, respectively, during 1 month before and 1 month after single gamma-irradiations with 25 cGy doses. Are established early (through 24 h after an irradiation) cytologic and genotoxic reactions in leukocytes of blood, and also hematological, biochemical reactions and changes of the content and frame DNA of leukocytes of blood in 30 day after an irradiation? Features of action of low doses of radiation in a combination to ions of mercury and their combination with a preparation of selenium are revealed. Communication of postradiating effects--changes of indexes of DNA of leukocytes of blood with reduction of life expectancy of animals is shown. The data are received, permissive to draw the conclusion, that the preparation of selenium in utilised concentrations, essentially not influencing on lifetime at low doses of radiation--mercury affecting is capable to improve quality of life of elderly animals PMID- 15754960 TI - [Clinical parameters of the course of metabolic syndrome in advanced age]. AB - The proposed principles of classification of metabolic syndrome are critically considered, as they do not take into account peculiarities of its clinical course in patients of advanced and senile age. Results of examination are presented of two groups of patients of the cardiological profile, who were long observed under conditions of a departmental outpatient clinic. In the first group, evaluated were the incidence and the degree of severity of symptoms characterizing disturbances in systems of carbohydrate and lipid metabolism, peculiar to metabolic syndrome, as well as the incidence and severity of arterial hypertension and obesity. It has been established that these symptoms are more frequent in various combinations as a picture of the "incomplete" syndrome and have different incidence in the older and younger age groups. Among patients with the "complete" set of characters of the metabolic syndrome there also takes place the statistically significant polymorphism of its clinical manifestations, which allows suggesting effects of mechanism of compensation and mutual aggravation. Peculiarities of manifestations of symptoms of the metabolic syndrome in patients of the elderly group make it possible to identify prognostically favorable and unfavorable forms of the course of the syndrome and, accordingly, to optimize the therapeutic and preventive programs. PMID- 15754962 TI - [Current tactics of the treatment of age-related macular degeneration]. AB - The survey offers an analysis on the current concepts of age-related macular degeneration's pathogeneses and treatment. PMID- 15754961 TI - [Diabetes mellitus in elderly: geroprotective and antidiabetic properties of delta-sleep induced peptide]. AB - Delta-sleep induced peptide (DSIP) has a wide range of positive regulator properties. It is known as antioxidant, immunomodulator, antistressor and other, that important as for diabetes, and for aging mechanisms. Aim of the study was to investigate effects of treatment by DSIP-containing drug "Deltaran" in elderly diabetic patients. MATERIALS AND METHODS: As the pilot study we examined 11 elderly diabetic patients (mean age 66.3 yr, BMI--28.5 kg/m2, duration of diabetes 12.5 yr). All patients receive 20 doses of DSIP-containing drug "Deltaran" daily. Tests was before and after 60 days after beginning treatment. Blood samples took before and after 60 minutes after standard carbohydrates breakfast. RESULTS: After treatment were observed improvement of the parameters of quality of life on SCAG scale as the impairment of recent memory (p=0.003), mood depression (p=0.019), emotional lability (p=0.002) and anxiety (p=0.000). Also were observed improvement of quality of sleep and felling between meals in treating patients (p=0.010). Another positive tendencies consist of decrease of BP (ns), increase of ABI (ns), increase of pallestesia (p=0.017), decrease of MAU frequency (p=0.011) and decrease of glycaemia levels after load (p=0.033). And also after treatment were observed decrease of basal and reactive levels in blood of growth hormone (p=0.034 and p=0.005 accord.), insulin (ns), cortisol (ns) and on the contrary increase of dehydroepiandrosterone sulfate levels (ns). Main positive change observed in patients received only diabetic diet, where was a decrease of insulin resistance and in patients with latent autoimmune diabetes (type LADA), where was a increase insulin secretion and decrease of daily dose of insulin requiring. CONCLUSIONS: The pilot trial indicate beneficial antidiabetic and geroprotective effects of DSIP-contaning drug "Deltaran" in elderly diabetic patients. PMID- 15754963 TI - [Community social gerontological work in the USA]. AB - This abstract represents community social gerontological programs and projects in the USA. There are many gerontological programs based on intergenerational approach. They involve the elderly and the young in joint activities and have good socio-psychological results. These programs and projects can be used in Russia, minding national and community traditions. PMID- 15754964 TI - [Internet resources in gerontology and social work with elderly in USA]. PMID- 15754965 TI - [Individual-typological distinctions of mental work capacity in aging]. AB - Investigated age and personal features of mental serviceability. It is revealed that psychological stability of the person renders stronger influence on mental serviceability than age. It is shown that mental serviceability depends not only on properties of temperament, but also on character traits. In this connection paramount value has formation of the emotional--strong-willed character traits, and also such properties of character, as self-esteem, advantage, independence, confidence of the abilities, social maturity that will promote preservation of mental serviceability at ageing. PMID- 15754966 TI - [Understanding aging]. AB - The essay reviews specific features of medical, biological, and biotechnological approaches to studying and understanding of biological phenomena exemplified with aging. PMID- 15754967 TI - [Comments on the article "Understanding aging" by Golubev AG]. PMID- 15754968 TI - Global patterns of isolation by distance based on genetic and morphological data. AB - The isolation-by-distance model predicts that genetic similarity between populations will decrease exponentially as the geographic distance between them increases, because of the limiting effect of geographic distance on rates of gene flow. Many studies of human populations have applied the isolation-by-distance model to genetic variation between local populations in a limited geographic area, but few have done so on a global level, and these few used different models and analytical methods. I assess genetic variation between human populations across the world using data on red blood cell polymorphisms, microsatellite DNA markers, and craniometric traits. The isolation-by-distance model provides an excellent fit to average levels of genetic similarity within geographic distance classes for all three data sets, and the rate of distance decay is the same in all three. These results suggest that a common pattern of global gene flow mediated by geographic distance is detectable in diverse genetic and morphological data. An alternative explanation is that the correspondence between genetic similarity and geographic distance reflects the history of dispersal of the human species out of Africa. PMID- 15754969 TI - Genetic influences on plasma cytokine variation in a parasitized population. AB - The soil-transmitted helminths are the most common helminthic infections, affecting about one-fourth of the world's population. There is a significant genetic component to susceptibility to infection with these organisms. Substantial changes in plasma cytokine levels are associated with helminthic infections, and there may be significant genetic components to this cytokine variation. Six plasma cytokine levels were assessed for 367 members of a single pedigree from the Jirel population of eastern Nepal. This population experiences moderate rates of infection with geohelminths. Sex, age, helminthic infection, infection with Giardia, and presence of a household latrine were considered as covariates in all analyses of the cytokine data. The analyses of the single Jirel pedigree revealed significant heritabilities for IFN-gamma (h2 = 0.654+/-0.096), TNF-alpha (h2 = 0.458+/-0.101), IL-2 (h2 = 0.583+/-0.101), IL-4 (h2 = 0.700+/ 0.095), IL-5 (h2 = 0.676+/-0.087), and IL-10 (h2 = 0.597+/-0.093). The ratios of IL-4 to IFN-gamma and of IL-10 to IFN-gamma were used as indicators of the degree of type 2 bias in immunological response; analyses of these variables indicated that approximately 40-60% of the variation (h2 = 0.400-0.577) in these derived measures of relative type 2/type 1 response is due to genetic factors. PMID- 15754970 TI - Genetic characterization of the population of Grande Comore Island (Njazidja) according to major blood groups. AB - The Comorian population is historically considered a blend of influences from African Bantus, Arabs, and possibly Austronesians. In this study we present the first genetic data on the current Comorian population. Serologic analysis of the six major blood group systems (ABO, RH, KEL, FY, JK, and MNS) was performed on 164 individuals from Grande Comore Island (Njazidja). In addition, Duffy genotypes were determined by polymerase chain reaction using allele-specific primers. Our findings establish a high frequency of the Fy(a- b-) phenotype (86%), presenting the same genetic background as in sub-Saharan Africa. Analysis of genetic frequencies, distances, and admixture with other populations indicates that African Bantus made the main contribution to the gene pool (73.2%+/-15.5%). The Arab contribution from the Arabian peninsula was smaller (24.2%+/-7%) and the Indonesian contribution was minor (2.6%+/-9%). The major Bantu contribution was commensurate with the Bantu cultural influence. The contribution from the Arabian peninsula seemed in relation to its permeating religious and linguistic influence. As with the language, the Indonesian contribution to the Comorian gene pool was small. These results are in agreement with historical, sociological, and linguistic data. PMID- 15754971 TI - Characterization of admixture in an urban sample from Buenos Aires, Argentina, using uniparentally and biparentally inherited genetic markers. AB - In this study we analyzed a sample of the urban population of La Plata, Argentina, using 17 mtDNA haplogroups, the DYS 199 Y-chromosome polymorphism, and 5 autosomal population-associated alleles (PAAs). The contribution of native American maternal lineages to the population of La Plata was estimated as 45.6%, whereas the paternal contribution was much lower (10.6%), clearly indicating directional mating. Regarding autosomal evidence of admixture, the relative European, native American, and West African genetic contributions to the gene pool of La Plata were estimated to be 67.55% (+/-2.7), 25.9% (+/-4.3), and 6.5% (+/-6.4), respectively. When admixture was calculated at the individual level, we found a low correlation between the ancestral contribution estimated with uniparental lineages and autosomal markers. Most of the individuals from La Plata with a native American mtDNA haplogroup or the DYS199*T native American allele show a genetic contribution at the autosomal level that can be traced primarily to Europe. The results of this study emphasize the need to use both uniparentally and biparentally inherited genetic markers to understand the history of admixed populations. PMID- 15754972 TI - Genetic diversity in Tunisia: a study based on the GM polymorphism of human immunoglobulins. AB - The GM polymorphism of human immunoglobulins is analyzed in three Berber populations of southern Tunisia and compared to other GM data. Genetic diversity among Tunisian populations is higher than that among Europeans but does not exhibit any significant geographic or linguistic structure. This result suggests a complex pattern of genetic differentiation. PMID- 15754973 TI - Genetic diversity and relationships among the tribes of Meghalaya compared to other Indian and Continental populations. AB - The autosomal AmpFLSTR markers validated and widely used for forensic applications are used in this study to examine the extent of diversity and genetic relationships among nine Meghalaya populations. Altogether, 932 chromosomes from 9 populations were analyzed using 9 tetrameric AmpFLSTR loci. The included populations were all seven subtribes of the Austro-Asiatic Mon-Khmer speaking Khasi and the neighboring Tibeto-Burman Garo. The Lyngngam, which are linguistically closer to the Khasi but are culturally intermediate between the Khasi and the Garo, are also included in the study. Although most of the microsatellite loci are highly polymorphic in each of these populations, the allele distributions are fairly uniform across the Meghalaya populations, suggesting relative homogeneity among them. Concurrent with this, the coefficient of gene differentiation (G(ST)) is observed to be low (0.026+/-0.002). This is naturally reflected in the lack of clear differentiation and clustering pattern of the Meghalaya tribes based on either geographic proximity or the historical or current affiliations of these tribes. Analysis of molecular variance (AMOVA) suggests no significant population structure. The structure analysis further suggests that, barring War-Khasi and Pnar, no other population shows any semblance of genetic identity. Even the position of the linguistically distinct Garo is not portrayed as separate from the Khasi. However, when comparable data from other Indian, Southeast Asian, and other continental populations were analyzed, the Meghalaya populations formed a compact cluster clearly separated from other populations, suggesting genetic identity of the Meghalaya populations as a whole. These results are concurrent with the hypothesis of a common and recent origin of these Meghalaya populations, whose genetic differentiation is overwhelmed by the homogenizing effect of continuous gene flow. PMID- 15754974 TI - Intra- and intercontinental molecular variability of an Alu insertion in the 3' untranslated region of the LDLR gene. AB - One-hundred three individuals from two Mongolian, two Siberian, and ten native American populations were studied in relation to a 340-bp sequence from an Alu insertion located in the 3' untranslated region of the LDLR gene. Seven haplotypes have been determined, and haplotype B1 was the most common, accounting for about half the sequences found. In general, diversity values are quite high, about 2.5 times higher than those found in other autosomal Alu sequences. Almost all (93%) of the variability occurs at the intrapopulation level, but the greatest among-group differentiation (6-8%) was found when we grouped in a single population all Native Americans plus Siberian Eskimos and Chukchi and compared them with Mongolians. This result is compatible with earlier mtDNA and Y chromosome suggestions of a single origin for the first colonizers of the American continent. With this nuclear locus it was not possible to broadly distinguish between Central and South American natives. No evidence of selection or marked demographic changes was obtained with these data. PMID- 15754975 TI - Apolipoprotein E polymorphism is related to plasma lipids and apolipoproteins in Mexican adolescents. AB - Previous studies in the Mexican population have failed to show an effect of apolipoprotein E (APOE) polymorphism on the lipid profile. The purpose of the present study was to determine the frequencies of APOE phenotypes, and their influence on lipid and apolipoprotein levels in a random sample of Mexican adolescents living in Mexico City. APOE polymorphism, fasting insulin levels, lipid levels, and apolipoprotein levels were determined in 420 adolescents. We found a high frequency of APOE*3 subjects (89.5%) and a low frequency of APOE*2 (3.0%) and APOE*4 (7.5%) subjects. The APOE*4 subjects (including APOE 4,3 and APOE 4,4) showed the highest concentrations of total cholesterol, low-density lipoprotein cholesterol, and apoB and the lowest high-density lipoprotein cholesterol levels, whereas carriers of the APOE*2 allele (APOE 3,2 and APOE 2,2) had the lowest values for total and low-density lipoprotein cholesterol and the highest concentrations of high-density lipoprotein cholesterol. No significant differences in triglyceride and insulin levels among subjects with different APOE polymorphisms were observed. Unlike previous studies in the Mexican population, our results show that lipid and lipoprotein levels are under the influence of APOE polymorphism. As in whites, APOE*4 may be a cardiovascular risk factor in the Mexican population. PMID- 15754977 TI - Worldwide variation in life-span sexual dimorphism and sex-specific environmental mortality rates. AB - In all human populations mean life span of women generally exceeds that of men, but the extent of this sexual dimorphism varies across different regions of the world. Our purpose here is to study, using global demographic and environmental data, the general tendency of this variation and local deviations from it. We used data on male and female life history traits and environmental conditions for 227 countries and autonomous territories; for each country or territory the life span dimorphism was defined as the difference between mean life spans of women and men. The general tendency is an increase of life-span dimorphism with increasing average male-female life span; this tendency can be explained using a demographic model based on the Makeham-Gompertz equation. Roughly, the life-span dimorphism increases with the average life span because of an increase in the duration of expressing sex- and age-dependent mortality described by the second (exponential) term of the Makeham-Gompertz equation. Thus we investigated the differences in male and female environmental mortality described by the first term of the Makeham-Gompertz equation fitted to the data. The general pattern that resulted was an increase in male mortality at the highest and lowest latitudes. One plausible explanation is that specific factors tied to extreme latitudes influence males more strongly than females. In particular, alcohol consumption increases with increasing latitude and, on the contrary, infection pressures increase with decreasing latitude. This finding agrees with other observations, such as an increase in male mortality excess in Europe and Christian countries and an increase in female mortality excess in Asia and Muslim countries. An increase in the excess of female mortality may also be due to increased maternal mortality caused by an increase in fertility. However, this relation is not linear: In regions with the highest fertility (e.g., in Africa) the excess of female mortality is smaller than in regions with relatively lower fertility (e.g., in Asia). A possible explanation of this phenomenon is an evolutionary adaptation of women to the pressures of extremely high fertility by means of some reduction of their maternal mortality. PMID- 15754976 TI - Variations in APOE genotype distribution in children from areas with different adult cardiovascular disease mortality in Spain. AB - We investigate whether a varying distribution of the APOE genotype could help explain regional differences in ischemic heart disease (IHD) mortality in Spain. APOE genotypes were examined by PCR in 1,274 randomly selected healthy children from four Spanish regions with different adult IHD mortality rates (northwest and central Spain with low rates and southeast and southern Spain with high rates). In the population as a whole the prevalence of the higher risk APOE*3/*4 genotype is 16.8% and the prevalence of the APOE*4 allele is 10.1%. In northwest Spain the frequencies of the APOE*3/*4 genotype (12.9%) and of the APOE*4 allele (8.3%) are smaller than in the other regions. The southeast region shows statistically higher frequencies of the APOE*3/*4 genotype (22.5%) and of the APOE*4 allele (13.2%) than in the other regions or in the group as a whole. We can conclude that Spain is not homogeneous in terms of APOE genotype distribution. Although the prevalence of the APOE*4 allele is generally low, there are areas with higher prevalence of the APOE*4 allele and a higher incidence of adult IHD mortality. This allows us to conclude that in Spain this genetic determinant can be associated with IHD mortality in relatively isolated populations. PMID- 15754978 TI - Distribution of CCR5-delta32, CCR2-64I, and SDF1-3'A mutations in populations from the Brazilian Amazon region. AB - The frequency distribution of the CCR5-delta32, CCR2-64I, and SDF1-3'A alleles was studied in the urban population of Belem and in Afro-Brazilians, Amerindians, and Japanese immigrants in the state of Para, Brazil. The results suggest that Amerindians may be genetically more susceptible to HIV-1 infection and disease progression than the other human groups studied. PMID- 15754979 TI - Model-based segmentation of medical imagery by matching distributions. AB - The segmentation of deformable objects from three-dimensional (3-D) images is an important and challenging problem, especially in the context of medical imagery. We present a new segmentation algorithm based on matching probability distributions of photometric variables that incorporates learned shape and appearance models for the objects of interest. The main innovation over similar approaches is that there is no need to compute a pixelwise correspondence between the model and the image. This allows for a fast, principled algorithm. We present promising results on difficult imagery for 3-D computed tomography images of the male pelvis for the purpose of image-guided radiotherapy of the prostate. PMID- 15754980 TI - The hypervolume under the ROC hypersurface of "near-guessing" and "near-perfect" observers in N-class classification tasks. AB - We express the performance of the N-class "guessing" observer in terms of the N2 N conditional probabilities which make up an N-class receiver operating characteristic (ROC) space, in a formulation in which sensitivities are eliminated in constructing the ROC space (equivalent to using false-negative fraction and false-positive fraction in a two-class task). We then show that the "guessing" observer's performance in terms of these conditional probabilities is completely described by a degenerate hypersurface with only N-1 degrees of freedom (as opposed to the N2-N-1 required, in general, to achieve a true hypersurface in such a ROC space). It readily follows that the hypervolume under such a degenerate hypersurface must be zero when N > 2. We then consider a "near guessing" task; that is, a task in which the N underlying data probability density functions (pdfs) are nearly identical, controlled by N-1 parameters which may vary continuously to zero (at which point the pdfs become identical). With this approach, we show that the hypervolume under the ROC hypersurface of an observer in an N-class classification task tends continuously to zero as the underlying data pdfs converge continuously to identity (a "guessing" task). The hypervolume under the ROC hypersurface of a "perfect" ideal observer (in a task in which the N data pdfs never overlap) is also found to be zero in the ROC space formulation under consideration. This suggests that hypervolume may not be a useful performance metric in N-class classification tasks for N > 2, despite the utility of the area under the ROC curve for two-class tasks. PMID- 15754982 TI - A three-parameter mechanical property reconstruction method for MR-based elastic property imaging. AB - A reconstruction process featuring full parameterization of the three dimensional, time-harmonic equations of linear elasticity is developed and reconstructed property images are presented from simulation-based investigation. While interesting in its own right through the potential for increased adaptability of these reconstructive elastic imaging techniques, this study also presents a set of analysis tools used to study the poor convergence behavior found in the case of tissue like conditions (i.e. nearly incompressible materials). The choice of elastic properties for imaging in elastography research remains an open question at this point; the use of the stability and sensitivity based analytical methods described here will help to predict and understand the value and reliability of different parameterizations of elasticity imaging. Additionally, though results indicate significant work needs to be done to achieve effective multiparameter reconstructive imaging, the methods detailed here offer the promise of increased flexibility and sophistication in elastographic imaging techniques. PMID- 15754981 TI - Detection performance theory for ultrasound imaging systems. AB - A rigorous statistical theory for characterizing the performance of medical ultrasound systems for lesion detection tasks is developed. A design strategy for optimizing ultrasound systems should be to adjust parameters for maximum information content, which is obtained by maximizing the ideal observer performance. Then, given the radio-frequency data, image and signal processing algorithms are designed to extract as much diagnostically relevant information as possible. In this paper, closed-form and low-contrast approximations of ideal observer performance are derived for signal known statistically detection tasks. The accuracy of the approximations are tested by comparing with Monte Carlo techniques. A metric borrowed and modified from photon imaging, Generalized Noise Equivalent Quanta, is shown to be a useful and measurable target-independent figure of merit when adapted for ultrasound systems. This theory provides the potential to optimize design tradeoffs for detection tasks. For example it may help us understand how to push the limits of specific features, such as spatial resolution, without significantly compromising overall detection performance. PMID- 15754983 TI - Conjugate phase MRI reconstruction with spatially variant sample density correction. AB - A new image reconstruction method to correct for the effects of magnetic field inhomogeneity in non-Cartesian sampled magnetic resonance imaging (MRI) is proposed. The conjugate phase reconstruction method, which corrects for phase accumulation due to applied gradients and magnetic field inhomogeneity, has been commonly used for this case. This can lead to incomplete correction, in part, due to the presence of gradients in the field inhomogeneity function. Based on local distortions to the k-space trajectory from these gradients, a spatially variant sample density compensation function is introduced as part of the conjugate phase reconstruction. This method was applied to both simulated and experimental spiral imaging data and shown to produce more accurate image reconstructions. Two approaches for fast implementation that allow the use of fast Fourier transforms are also described. The proposed method is shown to produce fast and accurate image reconstructions for spiral sampled MRI. PMID- 15754984 TI - Accelerated penalized weighted least-squares and maximum likelihood algorithms for reconstructing transmission images from PET transmission data. AB - We present penalized weighted least-squares (PWLS) and penalized maximum likelihood (PML) methods for reconstructing transmission images from positron emission tomography transmission data. First, we view the problem of minimizing the weighted least-squares (WLS) and maximum likelihood objective functions as a sequence of nonnegative least-squares minimization problems. This viewpoint follows from using certain quadratic functions as surrogate functions for the WLS and maximum likelihood objective functions. Second, we construct surrogate functions for a class of penalty functions that yield closed form expressions for the iterates of the PWLS and PML algorithms. Due to the slow convergence of the PWLS and PML algorithms, accelerated versions of them are developed that are theoretically guaranteed to monotonically decrease their respective objective functions. In experiments using real phantom data, the PML images produced the most accurate attenuation correction factors. On the other hand, the PWLS images produced images with the highest levels of contrast for low-count data. PMID- 15754985 TI - Visible Korean human: improved serially sectioned images of the entire body. AB - The data from the Visible Human Project (VHP) and the Chinese Visible Human (CVH), which are the serially sectioned images of the entire cadaver, are being used to produce three-dimensional (3-D) images and software. The purpose of our research, the Visible Korean Human (VKH), is to produce an enhanced version of the serially sectioned images of an entire cadaver that can be used to upgrade the 3-D images and software. These improvements are achieved without drastically changing the methods developed for the VHP and CVH; thus, a complementary solution was found. A Korean male cadaver was chosen without anything perfused into the cadaver; the entire body was magnetic resonance (MR) and computed tomography (CT) scanned at 1.0-mm intervals to produce MR and CT images. After scanning, entire body of the cadaver was embedded and serially sectioned at 0.2 mm intervals; each sectioned surface was inputted into a personal computer to produce anatomical images (pixel size: 0.2 mm) without any missing images. Eleven anatomical organs in the anatomical images were segmented to produce segmented images. The anatomical and segmented images were stacked and reconstructed to produce 3-D images. The VKH is an ongoing research; we will produce a female version of the VKH and provide more detailed segmented images. The data from the VHP, CVH, and VKH will provide valuable resources to the medical image library of 3-D images and software in the field of medical education and clinical trials. PMID- 15754986 TI - Determination of mechanical and electronic shifts for pinhole SPECT using a single point source. AB - The effects of uncompensated electronic and mechanical shifts may compromise the resolution of pinhole single photon emission computed tomography. The resolution degradation due to uncompensated shifts is estimated through simulated data. A method for determining the transverse mechanical and axial electronic shifts is described and evaluated. This method assumes that the tilt of the detector and the radius of rotation (ROR) are previously determined using another method. When this assumption is made, it is possible to determine the rest of the calibration parameters using a single point source. A method that determines the electronic and mechanical shifts as well as the tilt has been previously described; this method requires three point sources. It may be reasonable in most circumstances to calibrate tilt much less frequently than the mechanical shifts since the tilt is a property of the scanner whereas the mechanical shift may change every time the collimator is replaced. An alternative method for determining the ROR may also be used. Lastly, we take the view that the transverse electronic shift and the focal length change slowly and find these parameters independently. PMID- 15754987 TI - A study on several machine-learning methods for classification of malignant and benign clustered microcalcifications. AB - In this paper, we investigate several state-of-the-art machine-learning methods for automated classification of clustered microcalcifications (MCs). The classifier is part of a computer-aided diagnosis (CADx) scheme that is aimed to assisting radiologists in making more accurate diagnoses of breast cancer on mammograms. The methods we considered were: support vector machine (SVM), kernel Fisher discriminant (KFD), relevance vector machine (RVM), and committee machines (ensemble averaging and AdaBoost), of which most have been developed recently in statistical learning theory. We formulated differentiation of malignant from benign MCs as a supervised learning problem, and applied these learning methods to develop the classification algorithm. As input, these methods used image features automatically extracted from clustered MCs. We tested these methods using a database of 697 clinical mammograms from 386 cases, which included a wide spectrum of difficult-to-classify cases. We analyzed the distribution of the cases in this database using the multidimensional scaling technique, which reveals that in the feature space the malignant cases are not trivially separable from the benign ones. We used receiver operating characteristic (ROC) analysis to evaluate and to compare classification performance by the different methods. In addition, we also investigated how to combine information from multiple-view mammograms of the same case so that the best decision can be made by a classifier. In our experiments, the kernel-based methods (i.e., SVM, KFD, and RVM) yielded the best performance (Az = 0.85, SVM), significantly outperforming a well-established, clinically-proven CADx approach that is based on neural network (Az = 0.80). PMID- 15754988 TI - Parameter distribution models for estimation of population based left ventricular deformation using sparse fiducial markers. AB - We present a method to estimate left ventricular (LV) motion based on three dimensional (3-D) images that can be derived from any anatomical tomographic or 3 D modality, such as echocardiography, computed tomography, or magnetic resonance imaging. A finite element mesh of the LV was constructed to fit the geometry of the wall. The mesh was deformed by optimizing the nodal parameters to the motion of a sparse number of fiducial markers that were manually tracked in the images through the cardiac cycle. A parameter distribution model (PDM) of LV deformations was obtained from a database of MR tagging studies. This was used to filter the calculated deformation and incorporate a priori information on likely motions. The estimated deformation obtained from 13 normal untagged studies was compared with the deformation obtained from MR tagging. The end systolic (ES) circumferential and longitudinal strain values matched well with a mean difference of 0.1 +/- 3.2% and 0.3 +/- 3.0%, respectively. The calculated apex base twist angle at ES had a mean difference of 1.0 +/- 2.3 degrees. We conclude that fiducial marker fitting in conjunction with a PDM provides accurate reconstruction of LV deformation in normal subjects. PMID- 15754990 TI - Soft tissue differentiation using multiband signatures of high resolution ultrasonic transmission tomography. AB - In this paper, we are interested in soft tissue differentiation by multiband images obtained from the High-Resolution Ultrasonic Transmission Tomography (HUTT) system using a spectral target detection method based on constrained energy minimization (CEM). We have developed a new tissue differentiation method (called "CEM filter bank") consisting of multiple CEM filters specially designed for detecting multiple types of tissues. Statistical inference on the output of the CEM filter bank is used to make a decision based on the maximum statistical significance rather than the magnitude of each CEM filter output. We test and validate this method through three-dimensional interphantom/intraphantom soft tissue classification where target profiles obtained from an arbitrary single slice are used for differentiation over multiple other tomographic slices. The performance of the proposed classifier is assessed using receiver operating characteristic analysis. We also apply our method to classify tiny structures inside a bovine kidney and sheep kidneys. Using the proposed method we can detect physical objects and biological tissues such as styrofoam balls, chicken tissue, calyces, and vessel-duct successfully. PMID- 15754989 TI - Structural group classification technique based on regional fMRI BOLD responses. AB - This paper presents a new multigroup classification method based on subtle differences in regional brain activity during the completion of a functional magnetic resonance imaging (fMRI) challenge paradigm. Classification is performed based on features derived from BOLD time intensity curves in selected regions of interest (ROI). For each ROI, a mean time intensity curve [called mean regional response (MRR)] is calculated from realigned and normalized datasets. The overall subject performance is characterized with a vector of features obtained using nonlinear modeling of all subject's MRRs with a mixture of time shifted Gaussian functions. The classification is performed in the reduced-dimension optimal discrimination space, obtained through canonical transformations of original feature space. In order to demonstrate feasibility of the proposed method, classification of three groups of subjects is presented. The three groups are defined as heavy marijuana smokers after 24 hours of abstinence, heavy marijuana smokers after 28 days of abstinence, and healthy nonusing controls. The proposed method can be useful as an analytic tool for the discrimination of different groups of subjects based on temporal features of functional magnetic resonance imaging activation. PMID- 15754992 TI - Mean curvature mapping for detection of corneal shape abnormality. AB - Corneal topography is used to measure the anterior surface of the cornea. It is conventionally represented as radial slope, radial curvature, and elevation. In this paper, we introduce the application of mean curvature mapping as an alternative representation of the corneal topography. The purpose is to improve the detection of keratoconus and other diseases characterized by local increase in corneal curvature. Both simulated keratoconic cornea and real keratoconus data exported from the corneal topography system were analyzed. Four representations of corneal topography were generated and compared. It was found that mean curvature mapping provided the most precise cone location in simulated keratoconus. In both actual and simulated keratoconus cases, the appearance of the cone-like distortion is more consistent on mean curvature maps. Mean curvature mapping may improve the detection and localization of corneal shape abnormalities. PMID- 15754991 TI - Robust anisotropic Gaussian fitting for volumetric characterization of pulmonary nodules in multislice CT. AB - This paper proposes a robust statistical estimation and verification framework for characterizing the ellipsoidal (anisotropic) geometrical structure of pulmonary nodules in the Multislice X-ray computed tomography (CT) images. Given a marker indicating a rough location of a target, the proposed solution estimates the target's center location, ellipsoidal boundary approximation, volume, maximum/average diameters, and isotropy by robustly and efficiently fitting an anisotropic Gaussian intensity model. We propose a novel multiscale joint segmentation and model fitting solution which extends the robust mean shift-based analysis to the linear scale-space theory. The design is motivated for enhancing the robustness against margin-truncation induced by neighboring structures, data with large deviations from the chosen model, and marker location variability. A chi-square-based statistical verification and analytical volumetric measurement solutions are also proposed to complement this estimation framework. Experiments with synthetic one-dimensional and two-dimensional data clearly demonstrate the advantage of our solution in comparison with the gamma-normalized Laplacian approach (Linderberg, 1998) and the standard sample estimation approach (Matei, 2001). A quasi-real-time three-dimensional nodule characterization system is developed using this framework and validated with two clinical data sets of thin section chest CT images. Our experiments with 1310 nodules resulted in (1) robustness against intraoperator and interoperator variability due to varying marker locations, (2) 81% correct estimation rate, (3) 3% false acceptance and 5% false rejection rates, and (4) correct characterization of clinically significant nonsolid ground-glass opacity nodules. This system processes each 33-voxel volume of-interest by an average of 2 s with a 2.4-GHz Intel CPU. Our solution is generic and can be applied for the analysis of blob-like structures in various other applications. PMID- 15754993 TI - Immunohistochemistry in assessment of molecular pathogenesis of cervical carcinogenesis. AB - Concerning the prediction of HPV-associated cervical disease, several importance issues are related both to the management of women with diagnosed CIN and those with cervical cancer. Oncogenic HPVs are capable of contributing to the development of malignant phenotype by several different mechanisms, most of which seem to be closely interrelated. Because of the fact that these molecular interactions are mediated by proteins, the logical strategy to dissect the complex molecular pathways is to study the functions of these proteins, utilising the capabilities of immunohistochemistry (IHC). IHC offers practically unlimited possibilities to study any target molecules, against which a monoclonal or polyclonal antibody can be raised. This review describes the IHC-based strategies used by this author to assess the molecular pathogenesis of cervical cancer and its precursors in a number of large-scale prospective cohort studies conducted during the past 25 years. In the ongoing HPV-PathogenISS study, 13 different markers are being tested to evaluate their predictive value in distinct viral events, e.g. persistence or clearance of high-risk HPV in women treated for CIN. Apart from getting new insights into the molecular pathogenesis of HPV-associated cervical carcinogenesis, we anticipate the disclosure of individual markers, a set of markers, or an expression profile of any such marker sets that would be of clinical value as predictors of disease outcome in cervical carcinogenesis. PMID- 15754994 TI - Possibilities and limitation of endoscopic procedures in oncological gynaecology. AB - In cases of gynaecological cancer, the use of endoscopic techniques, in spite of becoming more frequent, is very controversial. Mainly, the potential risk of dissemination, which worsens the patient prognosis and leads to consequent adjuvant therapy is of concern. Based on data from the literature the authors present the advantages and disadvantages of using hysteroscopy in patients suffering from endometrial malignancy. According to the references and personal experience, the diagnostic and therapeutic value of laparoscopy in cases of cervical, endometrial and ovarian cancer are discussed. PMID- 15754995 TI - Restaging in gynaecological cancers. AB - Regardless of recent technical developments in the scientific arena, stage is still the most important prognostic factor in gynaecological cancers. Surgical staging is performed in all types of gynaecologic cancers except for cervical cancer. Adjuvant therapies that contribute to survival are planned in the light of information obtained from staging procedures. Therefore, necessary information for further therapeutic management should be revealed by the end of surgical staging. A staging surgery that is not completed for any reason will not only deprive the patient of necessary treatments, but can also cause administration of unnecessary adjuvant treatments. This is especially important, given the undesired effects and cost of both chemotherapy and radiotherapy. A particularly relevant case in point is tumours that look like early stage; this is because upstaging up to 30% has been reported in ovarian and endometrial cancers. As for vulvar cancer, clinical staging has been reported to lead to about 15% over diagnosis in comparison to surgical staging. Thus, the first step in all gynaecological cancers, except cervical cancer, should be to perform surgical staging when possible and unveil all surgical-pathological prognostic factors in the light of data obtained. Accordingly, restaging surgery should be considered in all cases that had incomplete staging. However, care should be taken to evaluate the benefits to be reaped together with the operative morbidity risk associated with the restaging procedure. This will both ensure accurate planning of postoperative treatment and provide a universal standard of approaching cancer patients and their treatments. PMID- 15754996 TI - Intratumoral oxygenation of invasive squamous cell carcimoma of the vulva is not correlated with regional lymph node metastasis. AB - INTRODUCTION: Tumour hypoxia has been found to be associated with tumour aggressiveness. Our primary aim was to explore the relationship between pretreatment tumour oxygenation in primary vulvar carcinoma and nodal status. Our secondary objective was to assess if there was a relationship between the clinical and biological variables. METHODS: 20 women with ISCC of the vulva were assessed with pretreatment primary tumour oxygenation with an Eppendorf pO2 probe. Patients underwent standard surgical management. Pathological assessment of the primary and nodal tissues was then performed. Primary tumour specimens were also stained for microvessel density and carbonic anhydrase IX. The relationship between smoking, preoperative Hgb, tumour CAIX expression, MVD, and Eppendorf pO2 measurements vs nodal metastasis and between these clinical and biological variables was assessed. RESULTS: Seven patients had positive lymph nodes, 13 had negative nodes. While neither current smoking status, tumour size, tumour oxygen measurements, MVD and CAIX expression correlated with metastatic nodal disease, a low preoperative Hgb correlated with pathological nodal status (p < 0.027). CONCLUSIONS: Although this analysis failed to demonstrate a strong correlation between various measures of tumour oxygenation with nodal metastasis, it may be due to the small number of patients. Only preoperative anaemia is correlated with nodal metastasis in early ISCC of the vulva. PMID- 15754997 TI - Primary lymphoma of the cervix uteri: a diagnostic challenge. Report of two cases and review of the literature. AB - BACKGROUND: Primary non Hodgkin's lymphoma (NHL) of the cervix and vagina is uncommon; the incidence of uterine lymphoma is estimated to be less than 0.5% of all NHL. Patients regularly present with vaginal bleeding. The diagnosis is made on biopsy but this can be difficult on small samples which may not be representative of the lesion. Immunohistochemical analysis and often molecular techniques are required to confirm the diagnosis. CASES: We report two cases of primary diffuse large B-cell lymphoma of the cervix. In the first case, the diagnosis could only be made on repeat biopsies. The second case presented as a cervical polyp. CONCLUSION: Gynecologists should be aware of this rare clinical entity in order to apply the proper treatment. PMID- 15754998 TI - Human papillomavirus infection in relation to mild dyskaryosis in conventional cervical cytology. AB - PURPOSE OF INVESTIGATION: To establish the prevalence and distribution of high risk human papillomavirus (HPV) genotypes in Slovene women with repeat mild dyskaryosis, and to evaluate three molecular methods for the detection of HPV that could be used as a complementary method to cervical cytology. METHODS: In this prospective study 148 women with three subsequent cervical cytologic tests within two years showing mild dyskaryosis were enrolled. HPV infection was determined using three molecular tests: Hybrid Capture II and two variants of polymerase chain reaction (PCR-PGMY11/PGMY09 and PCR-CPI/CPIIG). RESULTS: HPV was detected in 17 of the 45 women aged < or =30 years and in 21 of the 103 women aged >30 years (37.8% vs 20.4%, p = 0.04). The most common genotype was HPV 16 detected in eight (21.1%) women, the next were HPV 53 and HPV 51, each detected in five (13.2 %) women. The three molecular methods matched in 92.9%. CONCLUSION: Low prevalence of HPV infections indicates that cervical screening programmes in Slovenia are overburdened with mild dyskaryosis. Repeat cytology is not reliable; HPV testing might be useful as a complementary method. PMID- 15754999 TI - Evaluation of soluble tumour necrosis factor alpha receptors p55 and p75 in ovarian cancer patients. AB - PURPOSE: Evaluation of serum TNFalpha receptor 1 (p55) and 2 (p75) concentrations preoperatively in patients with ovarian masses. METHODS: Estimation by ELISA assay in 51 women with ovarian cancer and 16 healthy controls. Mean values and correlations with CA-125, tumour volume index, morphological score, pathological finding and cytoreduction were estimated. RESULTS: Mean concentrations of p55 and p75 in cancer patients were 2006 +/- 1030 pg/ml and 2849 +/- 1092 pg/ml, respectively, whereby for controls 1323 +/- 291 pg/ml and 2386 +/- 475 pg/ml, respectively. The area under the ROC curve for CA-125, p55 and p75 for cancer (FIGO Stages I-IV) were: 0.85 (95% CI 0.75-0.92), 0.73 (95% CI 0.60-0.83) and 0.65 (95% CI 0.50-0.77), respectively. Serum p55 correlated with morphological ultrasound score and CA-125 but not with FIGO stage, tumour grade or tumour volume index. No correlations of p75 with these parameters were observed. CONCLUSION: Estimation of p55 and p75 provide little information in ovarian cancer patients and have poor detecting power. PMID- 15755000 TI - Epidemiological aspects of ovarian malignancies in North Sardinia in the period 1992-2001. AB - Malignant ovarian tumors have been continuously increasing in Western countries and represent the leading cause of death for gynecological cancer. In fact, the mortality for malignant ovarian tumors remains very high with a low percentage of 5-year survival in the advanced stage of disease. The aim of this study was to evaluate the incidence trend and epidemiological characteristics of malignant ovarian tumors in the Province of Sassari, Sardinia (Italy) in the period 1992 2001 and to report the variations in comparison to the 1974-1985 period. The analysis of our data regarding the period 1992-2001, if compared with those of the period 1974-85, showed an increase of malignant ovarian tumors which triplicated achieving an incidence of 11.99/100,000 vs 4.27/100,000. The analysis of our epidemiologic data showed an increase of the age of first diagnosis (mean 60.9 years for epithelial ovarian tumors), the occurrence of the cancer in women at low socio-economic levels and a family history of cancer among the patients with malignant ovarian tumors. These data suggest that both local environmental factors combined with genetic characteristics play a role in the pathogenesis of ovarian tumors. The genetic characteristics could be of particular interest because Sardinia has been through the centuries a geographical area with little population migration. The marked increase in the incidence of ovarian tumors in the last several years points out the need to organize systematic screening by ultrasonography in our population. PMID- 15755001 TI - An ultrasonographic morphological index for prediction of ovarian tumor malignancy. AB - PURPOSE OF INVESTIGATION: A newly created ultrasonographic scale called the Poznan index as well as scales already well known (introduced by Sassone, De Priest and Lerner) were compared in our group of patients. METHOD: A morphological index was based on seven sonographic ovarian tumor features. Examinations on 686 patients were evaluated. Comparison of prognostic values of the Poznan index with other applied morphological indices in our group of patients was based on the area under receiver operating characteristic (ROC) curves. RESULTS: The cut-off level of the new index is 8 points. The new morphological index has a specificity of 77.0%, and negative and positive predictive values of 90.7% and 69.1%, respectively. It has a sensitivity of 86.7% and accuracy of 80.6%. The Poznan index proved its usefulness and superiority (AU ROC = 0.89). CONCLUSION: Using this morphological index it is possible to make a precise prognosis of ovarian tumor malignancy. It also makes it possible to make the right decision concerning the manner of surgical treatment. PMID- 15755002 TI - Endometrial cancer in polyps: a clinical study of 27 cases. AB - PURPOSE OF INVESTIGATION: To review risk factors, clinical presentation, diagnostic methods, and histopathologic findings in 27 cases of endometrial cancer in polyps. METHODS: A descriptive, retrospective study of 204 consecutive patients with endometrial carcinoma who were diagnosed at our institution between June 1998 to June 2001. Endometrial cancer arising in polyps occurred in 27 patients (13.2%) and accounted for 1.8% of 1492 endometrial polyps diagnosed during this period. RESULTS: Patients had a mean age of 62 years. All except one woman were postmenopausal. Three breast cancer patients were currently given tamoxifen. Metrorrhagia was the presenting symptom in 74% of cases, although 22% of patients were asymptomatic at the time of diagnosis. Ultrasonography performed in 22 patients showed images compatible with an endometrial polyp in 50% of cases, myoma in 5%, and inconclusive findings in 45%. The median endometrial thickness was 11 mm (range 4-33 mm). Diagnosis was made by aspiration-biopsy in 13 patients and by hysteroscopic endometrial sampling in 13 (in one patient endometrial carcinoma was incidentally found in the surgical specimen). All patients were in FIGO Stage IA. Endometrioid carcinoma was found in 81.5% of cases. Retroperitoneal metastases were not found in 25 patients undergoing pelvic lymphadenectomy, nor neoplastic growth in the specimens of six polypeptomies performed during hysteroscopy. All patients are free of relapse after a mean follow-up of 30 months. CONCLUSIONS: Postmenopausal women with endometrial polyps diagnosed by ultrasonography should undergo directed biopsies under hysteroscopic vision. The present series confirms the good prognosis of endometrial cancer in polyps. PMID- 15755003 TI - Risk factors and prognostic factors in patients with double primary cancer: epithelial ovarian cancer and breast cancer. AB - BACKGROUND AND OBJECTIVE: The most important known risk factor for ovarian cancer is the BRCA1-2 mutation, which is clinically often manifested through a positive family history of cancer of the breast and/or ovary. Whether other risk factors and prognostic factors in women with a positive family history of cancer of the breast and/or ovary and/or with BRCA1-2 mutation are important remains to be elucidated. Recent studies have shown that in the double primary breast and ovarian cancer (DPBOC), BRCA1-2 mutation is present in at least 86% of cases. Therefore, the group of patients with DPBOC, especially with epithelial ovarian cancer and breast cancer, is the most suitable for such an analysis. The aim of this study was to verify the hypothesis that, in this group, some other risk factors, in addition to a specific family history of cancer, as well as unfavourable pathomorphological prognostic factors, are more expressed than in a control group of patients with sporadic epithelial ovarian cancer only. METHODS: We compared the study group of 31 patients with DPBOC (epithelial ovarian cancer) to a control group of 62 patients with a single, sporadic epithelial ovarian cancer and negative specific family history. The data were obtained from the Cancer Registry of Slovenia and from clinical records. For every patient, we filled-in a protocol and analysed the data, comparing other risk factors in addition to specific family history and prognostic, clinical, and pathomorphological factors. Statistical analysis was performed using descriptive statistics, chi-square test and t-test. Multivariate analysis was also planned, but the necessary conditions were not met. RESULTS: In the study group, we found a higher percentage of positive non-specific family histories than in the control group, but the difference was not statistically significant. No difference in procreative risk factors was observed between the groups. There was a higher percentage of borderline significance of women from the study group that developed ovarian cancer between 45 and 59 years of age. In the study group, ovarian cancer was significantly more often found at Stage I, although the groups did not differ in detection procedures. Also, we did not find any differences in the distribution of tumour grades or histologic tumour types. CONCLUSION: The results did not confirm our hypothesis, yet they indicated some differences between the groups regarding the risk factors for ovarian cancer. Regarding the prognostic factors, we even found a significantly higher percentage of Stage I epithelial ovarian cancer in the study group, with no difference in the mode of detection. Considering the results that are not typical of BRCA-related cancer (what double primary cancer of the ovary and breast is supposed to be) and previous reports, we find it more likely that the patients with BRCA1-2 mutations represent only a subgroup within the group of patients with double primary breast and ovarian cancer. PMID- 15755005 TI - Ovarian cystic teratoma with primary epithelial cell melanoma. AB - We report a rare case of malignant melanoma arising in a cystic teratoma of the ovary occurring in a 60-year-old woman who died in four months despite the combined treatment administrated (surgery and chemotherapy). Diagnosis of ovarian melanoma was confirmed by immunohistochemical positivity to S-100 protein and HMB 45. There was no evidence of extra-ovarian primary melanoma on clinical examination; therefore the diagnosis was primary ovarian melanoma. Melanoma metastases were detected on the uterus, the right ovary, the omentum and in one of the three excised left external iliac lymph nodes. A review of the literature is analyzed and discussed. PMID- 15755004 TI - The relationship between mutant p53 gene, DNA contents and conventional clinicopathological prognostic variables in cases with endometrial carcinoma. AB - PURPOSE OF INVESTIGATION: To determine whether p53 expression and DNA ploidy are related to traditional prognostic indicators in patients with endometrial cancer. METHODS: Tumor material (n=136) was analyzed regarding flow cytometric DNA ploidy and immunohistochemical p53 expression. Pearson's correlation, Fisher's exact test, Cox's regression analysis and the Kaplan-Meier survival test were used, as appropriate. RESULTS: P53 overexpression and DNA ploidy were higher in patients with nonendometrioid histology, FIGO advanced stage, poor grade, positive peritoneal cytology, lymphovascular space invasion (LVSI) and lymph node involvement (LNI). Histologic subtype, stage, grade, LVSI, LNI, tumor recurrence and overall survival rate correlated with p53 and DNA ploidy. No association of depth of myometrial invasion and age with p53 and DNA ploidy was observed. P53 was related to DNA ploidy. Of the factors analyzed, histologic subtype and myometrial invasion were found to be most important independent determinants of recurrence. Utilizing survival as the endpoint for multivariate analysis, when considering p53 and DNA ploidy together, histologic subtype, stage, peritoneal cytology, LNI and DNA ploidy were independent prognostic indicators. CONCLUSION: p53 expression and DNA ploidy were related to histologic subtype, FIGO stage, grade, LVSI, LNI, peritoneal cytology, tumor recurrence and overall 5-year survival. As compared to p53, DNA ploidy was the stronger independent predictor factor for survival. Neither p53 nor DNA ploidy were significant independent factors for tumor recurrence when submitted to multivariate analysis in this study. However, since p53 or DNA ploidy were found to be significant factors in univariate analysis and were correlated with tumor recurrence, they could be useful factors in making prognoses. PMID- 15755006 TI - Diagnostic value of the PDD method in evaluation of vulvar lesions. AB - The purpose of this study was to evaluate the efficiency of photodynamic diagnostics (PDD), a method used in the diagnosis of vulvar lesions, and to determine its position and efficacy among diagnostic techniques generally used up to now. The projected purpose of the study in cases of vulvar lesions was realised by performing a detailed comparative analysis of sensitivity, specificity and diagnostic efficacy of PDD and vulvoscopy, as compared with histological evaluation of excised lesions. PMID- 15755007 TI - Preliminary experience with salvage weekly paclitaxel in women with advanced recurrent ovarian carcinoma. AB - PURPOSE OF INVESTIGATION: To assess the role of palliative chemotherapy with weekly paclitaxel in patients with recurrent ovarian cancer. METHODS: Thirty-two patients with paclitaxel- and platinum-resistant ovarian cancer were treated with weekly paclitaxel at 80 mg/m2 as a 1-hour intravenous infusion weekly for six weeks every eight weeks (1 cycle). This schedule was considered to be given for three cycles. Evaluation of radiographically measurable disease was used in the assessment of response. CA-125 was used to classify responses only in the absence of a measurable lesion. RESULTS: Thirty-two patients were all assessable for response. Of these, nine patients (28.1%) achieved a partial response and one patient achieved a complete response, leading to an overall response rate of 31.2%. Stable disease occurred in six patients (18.8%), and 16 patients (50%) had progressive disease. Nine patients died of progressive disease while on treatment. The median survival for the entire group was 10.5 months (range 2.5-22 months). Grade 3 or 4 leukopenia and neutropenia occurred in eight and six patients, respectively. Four of these patients developed febrile neutropenia without infection. Grade 1 and 2 peripheral neuropathies were observed in 50% of the patients without causing any premature drop out. Severe (grade 3 or 4) peripheral neuropathy was not observed. There were 11 patients with grade 1 or 2 myalgias. CONCLUSION: Weekly paclitaxel regimen is well tolerated with acceptable toxicity. The favorable toxicity profile and the encouraging antitumor activity observed in this study makes this regimen an option for the salvage treatment of patients with recurrent ovarian cancer. PMID- 15755008 TI - Quantitative telomerase activity in malignant, benign and normal gynecological tissues. AB - OBJECTIVE: The aim of this study was to evaluate quantitative telomerase activity in malignant, benign and normal gynecological tissue samples by using the Telomerase-PCR ELISA kit, and to determine a cut-off level for malignancy by this quantitative method. MATERIALS AND METHODS: Fifty gynecological tumors, 27 benign gynecological disorders and 29 normal tissues were analyzed by the Telomerase-PCR ELISA kit. All tissues were confirmed by a pathologist. A ROC (receiver operator characteristic) curve was drawn to determine a threshold level best discriminating malignant tissues from benign pathologies and normal tissues. Telomerase activity was compared in malignant, benign and normal tissues. RESULTS: The mean level of telomerase activity of the malignant tumor samples (1.03 +/- 0.53 units) was significantly (p < .001) higher than the normal tissues (0.13 +/- 0.07 units) and benign pathologies (0.37 +/- 0.25 units). The cut-off point to differentiate malignant samples from benign samples was set at 0.42 units, where the sensitivity was 93.8% and the specificity was 89.3%. Positive predictive value was 84% and negative predictive value was 89.3%. There was a significant difference in telomerase activity between malignant, benign and normal tissues within each histological group. CONCLUSION: In this preliminary study, the telomerase-PCR ELISA method was found to have a high sensitivity and specificity to differentiate malignant gynecological tissues from benign tissues. PMID- 15755009 TI - Granzyme B as a prognostic marker of cervical intraepithelial neoplasia. AB - Granzyme B (GrB) is a serine protease synthesized in T lympocytes (CTL), released after T-cell activation resulting from exogenous stimulation. With perforin, GrB discharges apoptotic signals to a target cell and therefore constitutes a marker to identify activated CTL. We aimed to quantify GrB expression by immunohistochemistry staining in 12 tissue fragments of cervical carcinoma, 33 cervical intraepithelial neoplasias treated by LLTEZ and nine cervical pieces without disease. Activated cytotoxic lymphocyte mean values (20 HPF-400x) in both epithelial and stromal pars were 7.11 cells in tissue without neoplasia, 33.45 cells in cervical intraepithelial neoplasia and 139.75 cells in carcinoma samples, with a statistical difference between them. Comparative analysis in the CIN group showed an expressive difference between cases with disease recurrence (19.28 cells) and without recurrence (37.26 cells). Thus, the relation between number of activated CTLs found at the moment of treatment and clinical evolution determined in this study, suggest GrB use as a prognostic marker. PMID- 15755010 TI - Value of glutathion-S transferase pi as a prognostic factor in epithelial ovarian carcinoma. AB - The association between glutathione S-transferase pi (GSTpi) and other clinicopathological parameters, response to chemotherapy and clinical outcome were investigated in chemotherapy naive epithelial ovarian cancer patients. Paraffin-embedded material from 55 patients were used for immunohistochemical analysis. All patients had received six cycles of cisplatinum-based chemotherapy and 41 of them were revalued by laparotomy. Pre- and post-chemotherapy GSTpi staining were detected in the cancer tissues of 18/55 (32.7%) and 5/14 (35.7%) patients, respectively. GSTpi expression was not associated with other clinicopathologic parameters. Of 17 patients with postoperative measurable residual disease clinical response was observed in 4/7 of GSTpi positive and in 9/10 GSTpi negative patients (p = 0.25). Pathologic complete response (pCR) was achieved in 5/8 of GSTpi positive and 11/22 of GSTpi negative cases (p = 0.69). There was no significant difference in overall survival and progression-free survival (PFS) according to initial GSTpi status. However the PFS of the five patients (median 22 +/- 5.9 months) who had postchemotherapy positive GSTpi was significantly shorter than the nine patients (10.0 +/- 2.19 months) who had negative GSTpi (p = 0.006). This difference was not observed in overall survival. These results suggest that initial immunohistochemical staining of GSTpi does not aid in the prediction of pCR and clinical outcome in patients with epithelial ovarian cancer. Nonetheless investigation of GSTpi expression after chemotherapy needs further evaluation. PMID- 15755011 TI - Expression of E-cadherin in squamous cell carcinomas of the cervix with correlations to clinicopathological features. AB - OBJECTIVE: To evaluate the expression of E-cadherin, a calcium-dependent cell adhesion molecule, in a retrospective analysis of paraffin-embedded tissue specimens of cervical squamous carcinoma and the relationship with histopathological differentiation and lymph node status. METHODS: In this study, we investigated by immunohistochemistry E-cadherin expression in ten normal cervical epithelia and 24 cervical invasive squamous carcinomas. RESULTS: Normal cervical squamous epithelium showed strong expression of E-cadherin at the membrane of the cell and intercellular junctions. In 24 tumors immunnostained by E-cadherin antibody, 11 (46%) showed preserved expression and 13 (54%) reduced expression. There was no significant correlation between E-cadherin expression and histological differentiation (p = 0.650, p = 0.294). In the status of lymph node metastasis, reduced expression of E-cadherin was seen in 11/15 (73%) with lymph node metastasis versus 2/9 (22%) without lymph node metastasis. There was a significant inverse correlation between E-cadherin expression and lymph node metastasis (p = 0.032). CONCLUSION: Reduced E-cadherin expression may be an important factor among a variety of biologic events that occur during the process of metastasis. However, this should be explored by a large scale study. PMID- 15755012 TI - Antiestrogenic therapy in breast cancer and endometrial modifications. AB - The aim of this retrospective study was to detect endometrial lesions in tamoxifen breast cancer users (menopausal state related). The meaning of genital bleeding during the treatment and the actual incidence of benign and malignant pathology of the endometrium related to length of treatment was also evaluated. Tamoxifen (TMX) is a nonsteroidal triphenylene derivate with clear antiestrogenic properties on the breast which is used as adjuvant treatment for breast cancer; potential adverse effects include endometrial lesions. Three hundred and sixty six breast cancer patients were enrolled in this study; 292 patients were treated with 20 mg/daily of TMX as adjuvant therapy and the remaining 74 did not receive therapy. All patients were subdivided in premenopausal and postmenopausal, asymptomatic and symptomatic groups. All patients underwent ultrasound scans (to examine endometrial thickness) and hysteroscopic examinations before treatment and after one, three and five years. Endometrial biopsy under direct hysteroscopic vision was systematically performed. The pathological histology reports were classified under polyps, simple hyperplasia, complex hyperplasia, atypical hyperplasia, and carcinoma. A higher incidence of endometrial pathology was found only in symptomatic postmenopausal TMX treated patients (27.2% vs 19.5%) between the third and fifth year of treatment. PMID- 15755013 TI - Vulvar invasive squamous cell carcinoma occurring in a young woman with systemic lupus erythematosus. AB - BACKGROUND: Although several studies have demonstrated a possible relationship between systemic lupus erythematosus (SLE) and non-Hodgkin's lymphoma, Hodgkin's lymphoma, leukemia and several solid tumors, it is still debatable whether SLE patients have an increased incidence of cancer overall. CASE: We describe a 25 year-old patient with SLE who developed invasive squamous cell carcinoma of the vulva. The patient underwent radical vulvectomy and bilateral groin sentinel lymph node dissection and until to date, one year after surgery, she is alive without evidence of recurrent disease. CONCLUSIONS: Only three cases of vaginal/vulvar cancer associated with SLE have previously been mentioned in the literature, but not described in detail. This is the first detailed case report in the literature of vulvar invasive squamous cell carcinoma occurring in a SLE patient. It can only be speculated that the SLE itself and/or the treatment with immunosuppressive drugs provoked malignant transformation and the development of vulvar squamous cell carcinoma in such a young patient. PMID- 15755014 TI - Primary squamous cell carcinoma of endometrium: a case report. AB - A case of primary squamous cell carcinoma of the endometrium (PSCCE) in a virgin treated with surgery--abdominal hysterectomy and bilateral salpingo-oophorectomy, and followed by whole pelvic radiation is presented. The tumor recurred 12 months later and the patient then underwent relaparotomy and excision of recurrent tumor of the pelvis; right hemicolectomy and rectosigmoidectomy were done due to recurrence and metastasis of the primary tumor. After re-operation, six courses of cisplatin and 5-fluorouracil were given intravenously for cytotoxic effects at 3-week intervals. The patient's disease progressed despite therapy, and she died three months after the last cycle or 21 months after the first diagnosis was made. PMID- 15755015 TI - Coexistence of brenner tumor and struma ovarii: case report. AB - BACKGROUND: There has been controversy regarding the histogenesis of Brenner tumors. It is generally accepted that Brenner tumors are derived directly from ovarian surface epithelium, which undergoes metaplasia to form the typical urothelial-like components, whereas some investigators assume that Brenner tumors arise from immature germ cells. CASE: We describe a well-documented case of the coexistence of struma ovarii regarded as a form of teratoma and Brenner tumor in the same ovary. Immunohistologically, not only columnar cells of thyroid follicles, but also transitional cells of Brenner nests were positive for thyroglobulin. CONCLUSIONS: In the present case, Brenner tumors and thyroid elements coexisted and were positive for thyroglobulin. While there is strong evidence that pure Brenner tumors originate mostly from the ovarian surface, at least Brenner tumors associated with teratomatous elements may have a germ cell origin. PMID- 15755016 TI - Laparoscopic uterine artery dissection in an undiagnosed endometrial stromal sarcoma. Case report. AB - We present a case of a 24-year-old woman, gravida 0, with menometrorrhagia and pelvic pain. A uterine hemorrhagic fibroid was diagnosed after ultrasound and magnetic resonance imaging (MRI). The endometrial biopsy was negative for malignancy. Laparoscopic sentinel lymph node sampling, lavage, and myometrial biopsy with negative results were performed before dissection of the uterine vessels. The final diagnosis of endometrial stromal sarcoma was made by myomectomy and hysterectomy one year later. This case should demonstrate the difficulty of making the right diagnosis of sarcoma before laparoscopic dissection of uterine vessels in patients with symptomatic fibroids. PMID- 15755018 TI - Pelvic retroperitoneal schwannoma presenting as a gynecologic mass: case report. AB - Schwannomas of the sacral plexus are retroperitoneal tumors, usually benign, that result from proliferation of perineural cells. They are rare, pelvic, well defined tumors that present infrequently as gynecologic masses. Misdiagnosis is not uncommon in these cases, since these masses are not often encountered in clinical practice and preoperative imaging methods can be only suggestive of the diagnosis. Furthermore, these masses are not often encountered in clinical practice. We report a case of a 28-year-old woman who presented with a pelvic mass and obscure clinical signs. Imaging methods suggested that this tumor was most probably an ovarian mass of embryonic origin. After dissection by laparotomy, the microscopic examination showed typical futures of a neurilemoma (ancient schwannoma). The goal of the operating intervention is to excise the tumor avoiding major trauma, thus cooperation between surgeons is indispensable. The patient has had no motor or sensory disturbances after surgery. PMID- 15755017 TI - Endometrial stromal sarcoma arising in extrauterine endometriosis: a case report. AB - We report a rare case of a 46-year-old woman developing endometrial stromal sarcoma (ESS) on the grounds of extrauterine endometriosis. The patient presented with symptoms of stenosis of the rectosygmoid colon. The tissue samples were submitted to histological and immunohistochemical analyses using antibodies for indirect staining. The trial showed multiple foci of endometriosis and mesenchymal malignant tissue described as ESS in the bowel wall, mesentery and in the remnants of the left adnexae. According to our findings, we suspect that ESS might have arisen in colon endometriosis. PMID- 15755019 TI - Primary ovarian leiomyosarcoma: a case report. AB - Primary ovarian leiomyosarcomas are extremely rare tumors that comprise less than 0.1% of all ovarian malignancies. We present a case of 62-year-old postmenopausal woman with a slightly enlarged right ovary and a Color Doppler sonography resistance index (RI) measuring 0.54. The patient, after being managed with surgery alone, is alive after 14 months without any evidence of disease. This is an unusual case in that primary ovarian leiomyosarcoma was diagnosed in the setting of a slightly enlarged irregular postmenopausal ovary with a concomitant intermediate RI value on color flow Doppler evaluation. A high index of suspicion may help prevent delay in the diagnosis of this rare neoplasm. PMID- 15755020 TI - Laparoscopic evaluation of metastatic ovarian cancer: a case report. AB - OBJECTIVE AND CASE: Both noninvasive and invasive methods have limited value in the diagnosis of metastatic ovarian cancer. We present a case with the initial complaint of abdominal distention in whom primary and metastatic tumor sites were safely diagnosed by using laparoscopy: a gastric tumor with ovarian metastasis. DISCUSSION: Diagnostic laparoscopy by the open technique provides a safe and effective diagnostic option in patients with metastatic ovarian cancer. PMID- 15755021 TI - [Urgent need for the standardization of semen analysis among clinical andrology laboratories]. PMID- 15755022 TI - [Is the semen analysis a reliable test?]. AB - The semen analysis is one, if not the most, important and widely used clinical laboratory test to evaluate the fertility potential of the male. However, recent reports have suggested that the semen analysis is unreliable. Quality control in the andrology laboratory is often seen as problematic, and many laboratories do not routinely employ QC procedures in semen testing. Quality assurance is an often overlooked and unappreciated aspect of overall quality laboratory performance. External proficiency testing programs in andrology are not universally accepted, and the results from the few programs currently available demonstrate huge variations between laboratories. Numerous different standards and criteria are being used by andrology laboratories, making it difficult if not impossible to compare results from one laboratory to another. However, reliable semen analyses can be obtained by following several recommendations: (1) all laboratories performing the semen analysis should adopt universally accepted performance standards and criteria, (2) all laboratories performing this test should participate in external proficiency testing programs, (3) andrology laboratories should implement effective internal quality control and quality assurance programs to ensure that the results reported are accurate and reproducible, and (4) physicians should only refer their patients to, or accept semen analysis results from, laboratories that have stringently followed these recommendations. PMID- 15755023 TI - [Quantitative morphometry of hyperplastic hypoechoic nodules in the prostatic outer gland]. AB - OBJECTIVE: To explore the morphotic characteristics of hypoechoic nodules in the outer gland of the prostate with benign METHODS: Twenty-two hyperplastic hypoechoic nodules in the prostatic outer gland were biopsied guided by prostatic hyperplasia. transrectal ultrasound. The hematoxylin-eosin (HE) staining and immunohistochemistry combined with computer assisted quantitative image analyses were adopted to examine the mean percentages of the area densities of stroma, epithelium, glandular lumen and smooth muscle cells. RESULTS: The area densities of stroma, epithelium, glandular lumen and smooth muscle cells were (72.52 +/- 13.14)%, (20.57 +/- 9.01)%, (6.85 +/- 4.51)% and (24.14 +/- 6.31)%, respectively. CONCLUSION: Hyperplastic hypoechoic nodules may develop in the outer gland as well as in the inner gland of the prostate, but the mean percentages of the components are different between the two kinds of nodules. PMID- 15755024 TI - [Effect of hypertension on cell proliferation and apoptosis in benign prostatic hyperplasia]. AB - OBJECTIVE: To evaluate the effect of hypertension on cell proliferation and apoptosis in benign prostatic hyperplasia (BPH). METHODS: The techniques of immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling(TUNEL) were used to detect the proliferative index (PI) and apoptotic index (AI) in both epithelium and stroma of simple BPH (40 cases) and BPH with hypertension (40 cases), respectively. RESULTS: In both the groups of simple BPH and BPH with hypertension, the value of PI was significantly higher than that of AI in epithelium and stroma (P < 0.05), and PI was closely related to the prostate volume (P < 0.05). As compared with the simple BPH group, PI was significantly increased in epithelium and stroma in the group of BPH with hypertension( P < 0.05), in which the disease history of hypertension was positively associated with PI in epithelium and stroma (p < 0.05). CONCLUSION: Hypertension, particularly persistent hypertension state, may enhance cell proliferation in both epithelium and stroma of BPH, and consequently lead to the enlargement of the prostate. PMID- 15755025 TI - [Growth hormone enhances regeneration of nNOS-containing nerve fibers in the aged rat penis]. AB - OBJECTIVE: To investigate the effects of growth hormone (GH) on the erectile function and the number of neuronal nitric oxide synthase (nNOS) -containing nerve fibers in the penis of aged rats. METHODS: Twenty-four aged male SD rats (18 months) were randomized into 2 groups: GH intervention group and control group. After four and eight weeks, a half of each group were selected and tested for erectile function after apomorphine (APO) injection and then sacrificed for the detection of nNOS-containing nerve fibers in the penis by streptavidin peroxidase conjugated method (SP method). RESULTS: After four weeks, the erectile function and the number of nNOS-containing nerve fibers showed no significant difference between the GH intervention group and the control group (P > 0.05). After eight weeks, the erection frequency was significantly different (P < 0.05) between the two groups, while the erection rate was not. The number of nNOS containing nerve fibers in the GH intervention group was significantly larger than that in the control group (P < 0.01). CONCLUSION: GH enhances the regeneration of nNOS-containing nerve fibers in the penis and improves the erectile function of the aged rat. PMID- 15755027 TI - [Expression of cyclooxygenase-2 in the testes and epididymides of adult male rats]. AB - OBJECTIVE: To investigate the expression of cyclooxygenase-2 (cox-2) in the testes and epididymides of adult male rats and its significance. METHODS: Immunohistochemical staining was used to detect the expression and localization of cox-2 in the testicle and epididymal tissues of 40 adult male SD rats. RESULTS: Strong cox-2 immunoreactivity was detected in the epididymides and testes of the rats. In the caput epididymides, cox-2 expressed mainly in the epithelial nuclei and partly in the cytoplasm. Cox-2 was also found positive in the testis nuclei and cytoplasm. CONCLUSION: Immunohistochemical staining is a fairly sensitive method for detecting cox-2 expression in the testes and epididymides of adult male rats. PMID- 15755026 TI - [Comparison between two methods for staining DNA of apoptotic spermatozoa]. AB - OBJECTIVE: To compare two fluorochrome staining methods for the assessment of sperm quality. METHODS: Washed sperm cells were incubated in 0, 0.15, or 15 micromol/L camptothecin (CAM), or 0.37 or 3.7 mmol/L genistein (GEN) at 37 degrees C for 4 hours. The sperm cells were analyzed for cycle-independent apoptosis and necrosis by single-stain compared with dual-stain fluorescence microscopy to contrast the relative effectiveness of these two approaches. RESULTS: The single-stain procedure could not detect the sperm viability differences. In contrast, the dual-stain procedure identified a dosage-dependent decrease in the viability and increased necrozoospermia after topoisomerase inhibitor CAM and GEN treatments. Apoptosis was 2-fold higher with topoisomerase inhibitor treatment. CONCLUSION: The two topoisomerase inhibitors were associated with increased apoptosis and dosage-dependent necrosis. The data suggested that the dual-stain combination Hoechst 33342/PI was more sensitive than the single Hoechst 33342 stain analysis and permitted quantitative analysis of the apoptosis and necrosis in sperm. PMID- 15755028 TI - [Expression and biology identification of the human epididymis-specific gene ESC42 in E. coli]. AB - OBJECTIVE: To provide materials for the study of the function of ESC42 protein specifically expressed in the human epididymis. METHODS: The ESC42 gene was amplified from the human epididymis cDNA library by PCR and then cloned into prokaryotic expression vector pGEX-4T-1, expressed and purified by recombinant DNA techniques. The specificity of ESC42 protein was identified by Western blot and MALDI-TOF-MS. The database was searched by Ms-Fit. RESULTS: The recombinant plasmid expressed a Mr 38 x 10(3) fusion protein in E. coli at a level of 30% of the total protein, and the purity was as high as 99%. The ESC42 protein was identified by ESC42 monoclonal antibody and its molecular weight was 11 978.12, tested by MALDI-TOF-MS. The peptide mass fingerprint analysis showed that the coverage rate of the sequence reached 48% with 100% matching. The motif scan in Prosite database reveal that ESC42 belonged to the beta-defensin family and had antibacterial activity. CONCLUSION: Obtaining high purity of rhESC42 protein may lay a foundation for the study of its functions. PMID- 15755029 TI - [Correlation between abnormal serum lipid and erectile dysfunction]. AB - OBJECTIVE: To achieve the correlation between abnormal serum lipid and male erectile dysfunction (ED). METHODS: Randomly selected were 200 male patients abnormal at least in one of the four laboratory items: total cholesterol (TC), triglycerides (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL). Their erectile function was evaluated by IIEF-5, and the correlation was studied between abnormal serum lipid and ED. RESULTS: The incidence of ED was 47% in these patients. Age, coronary heart disease (CHD), high serum glucose, benign prostatic hyperplasia (BPH), drug, hypertension were negatively, while HDL was positively correlated with erectile function. Age, CHD, BPH and TC/HDL ratio were risk factors of ED, while HDL was a preventive one. CONCLUSION: Hyperlipidemia is an important factor of ED. HDL and TC/HDL ratio are good predictors of ED. PMID- 15755030 TI - [Effects of polychlorinated biphenyl on bcl-2 and TGFbeta1 expression in rat testes]. AB - OBJECTIVE: To study the effects of polychlorinated biphenyl (PCB) on bcl-2 and TGFbeta1 expression in rat testes. METHODS: Forty male Wistar rats were divided into 4 groups at random: Group A (normal control), Group B (fed on 10(-8) mol/L PBC), Group C (feb on 10(-7) mol/L) and Group D (feb on 10(-6) mol/L). After three months, all the rats were killed, the animal model established, and observations made on the expression of bcl2 and TGFbeta1 in the rat testis using the optical microscope and immunohistochemical techniques. RESULTS: The damage to the structure of the testis was related to the dosage of PCB: the higher the dodage, the more serious the damage. PCB induced the expression of bcl-2 and TGFbeta1. The TGFbeta1 expression was significantly higher in the highest dosage group than in others (P < 0.01 ), and the bcl-2 expression was dramatically higher in Group C than in other groups (P < 0.01). CONCLUSION: PCB can cause injury in rat testes. PMID- 15755031 TI - [Microcirculation changes in the ventral prostates of rats after castration]. AB - OBJECTIVE: To study the microcirculation changes in the ventral prostates of rats after castration and the role of microcirculation during the apoptosis of prostatic cells. METHODS: Thirty-six male adult rats were randomized to 6 groups: one was taken as the control, while the other 5 underwent measurement of the microcirculation in vivo by a D95 physiological signal acquisition system 12 h, 24 h, 72 h, 7 d and 14 d respectively after castration, and then were perfused with Chinese ink to trace the microvessels of the prostates. RESULTS: The microcirculation of the rats'prostates changed dramatically following castration. The diameter and density of the microvessels, especially in the distant and mediate ducts of the prostates, decreased dramatically, and so did the bloodflow velocity. CONCLUSION: The microcirculation plays a role during the process of apoptosis of prostatic cells, and might be the mechanism of " apoptosis shift". PMID- 15755032 TI - [Effect of photodynamic therapy with metalloporphyrin compound on human prostate cancer PC-3 cells in vitro]. AB - OBJECTIVE: To investigate the effect of the photodynamic therapy (PDT) with the new water-soluble metalloporphyrin compound on human prostate cancer PC-3 cells in vitro and the anticancer mechanism of PDT. METHODS: The new water-soluble manganese, 5,10,15, 20-tetra (N-methyl4-pyridyl) porphinato (2-) tetraiodide salt, was synthesized. The PC-3 cells were treated with the compound of serial concentrations(0, 0.1, 1, 1.0 micromol/L) followed by irradiation of different dosages of visible light. The techniques of MTT and Annexin-V/propidium iodide double-labeled flow cytometry (FCM) were applied to measuring the inhibitory effect of the compound on the growth activity and apoptosis of the cells. RESULTS: When the metalloporphyrin compound concentration was within 10 micromol/L and the irradiation time was within 30 min, the water-soluble metalloporphyrin compound had a significant inhibitory effect on the proliferation of PC-3 cells and induced PC-3 cell apoptosis, and the effects depended greatly on metalloporphyrin concentration and illumination dosages. Higher concentrations and dosages induced the death of the majority of PC-3 cells. CONCLUSION: The PDT of the water-soluble metalloporphyrin compound followed by light irradiation has a distinctive killing effect on PC-3 cells in vitro, and the rates of proliferation inhibition and cell apoptosis are correlated with metalloporphyrin concentration and the dosages of light irradiation. The results suggest that the mechanism of metalloporphyrin PDT may be involved with the induction of apoptosis in human prostate cancer cells. PMID- 15755033 TI - [A clinical study of biological zinc for the treatment of male infertility with chronic prostatitis]. AB - OBJECTIVE: To evaluate the efficacy of biological zinc in the treatment of male infertility with chronic prostatitis (MICP). METHODS: Thirty-eight patients with MICP were treated with biological zinc. The zinc concentration in the semen and the seminal parameters were tested before and after using biological zinc. RESULTS: After treatment, the zinc concentration in the semen was increased markedly, and the semen liquefaction and the sperm motility were also improved in the patients who had received biological zinc supplementation as compared with those who had not (P < 0.05). CONCLUSION: It is suggested that biological zinc has the effect of increasing zinc concentration in semen, and the supplementation of biological zinc for one of the effective methods for the treatment of MICP. PMID- 15755034 TI - [Extraperitoneal laparoscopic radical prostatectomy: a report of 2 cases]. AB - OBJECTIVE: To probe into the operation method and clinical result of extraperitoneal laparoscopic radical prostatectomy. METHODS: Two male patients of prostate cancer underwent extraperitoneal laparoscopic radical prostatectomy. The main operation procedures proceeded under the extraperitoneal laparoscope, consisting of dissecting the prostate gland, cutting the bladder shank and the apix of the prostate gland, and then freeing the seminal vesicles followed by removing the prostate anteriorly. The final step was to connect the urethra and bladder neck. RESULTS: The operation time was 10 and 7 hours and blood loss was 1 000 and 500 ml respectively. The intestinal function resumed 24 hours after the operation. The catheter was removed 3 weeks after surgery and no complication was seen. CONCLUSION: Extraperitoneal laparoscopic radical prostatectomy is a good and least invasive method for local prostate cancer. PMID- 15755035 TI - [Correlation of age and sex hormones to the transition zone volume of the prostate]. AB - OBJECTIVE: To investigate the correlation of age and sex hormones to the total volume (TV) and transition zone volume METHODS: Eighty-two men were divided into two groups with the age of 60 as the dividing line. Prostatic TV (TZV and TZV were measured with transrectal ultrasound (TRUS) and the concentrations of total testosterone (T), free testosterone (fT) and estradiol (E2) were measured with radioimmunoassay. Exponential curve regression and one-factor analysis of variance (ANOVA) were used for statistical analysis. RESULTS: Age was significantly correlated with TV, TZV and the TZV/TV ratio, P < 0.01. There was a significant positive correlation between the concentrations of fT and TZV, a negative correlation between those of E2 and TZV (P < 0.01), but no correlation between those of T and TZV (P > 0.05). CONCLUSION: The significant acceleration of the TZV of the prostate after the age of 60 is related to sex hormones. PMID- 15755036 TI - [Effect of FK506 on the cavernous nerve regeneration after injury]. AB - OBJECTIVE: To investigate the effect of FK506 on the cavernous nerve regeneration after injury and to discuss its possible action mechanisms. METHODS: Fifty-four male adult Sprague-Dawley rats were randomized into three groups: Group 1 (sham control), Group 2 (unilateral cavernous nerve ablation), and Group 3 (unilateral cavernous nerve ablation with subsequent injection of FK506). Electrostimulation of the cavernous nerve was performed at 1 and 3 months after surgical injury. The intracavernous pressure was continuously detected and the rats were followed by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) staining to identify NOS in the penile nerve fibers of the penile shaft. RESULTS: At 1 month, the number of NOS-positive nerve fibers significantly decreased with no statistical difference among the three groups except the sham controls (P > 0.05). At 3 months, electrostimulation revealed greater maximal intracavernous pressure in Group 3 than in Group 2 (P < 0.01). Furthermore, the number of NOS positive nerve fibers showed a significant increase (P < 0.01), but not in Group 2 (P > 0.05). CONCLUSION: FK506 injection enhances the regeneration of cavernous nerves after injury and the recovery of erectile function in rats. PMID- 15755037 TI - [Transurethral prostatectomy with the bipolar plasma kinetic technique for benign prostate hyperplasia: a report of 297 cases]. AB - OBJECTIVE: To evaluate the effect and safety of transurethral prostatectomy with the bipolar plasma kinetic technique (PKRP) in the treatment of benign prostate hyperplasia(BPH). METHODS: Two hundred and ninty-seven BPH patients underwent transurethral prostatectomy with the bipolar plasma kinetic technique. The preoperative estimated weight of the prostate ranged from 35 g to 102 g, averaging 52 g. RESULTS: The operation lasted 40 approximately 65 min, averaging 51 min. The resected tissues weighed 40 approximately 80 g, averaging 46 g. During the operation no transurethral resection (TUR) syndrome occurred. The catheter was removed 4 approximately 5 days after the operation, all with fluent urination. The patients were followed up for 2 approximately 33 months. IPSS decreased from average 31.5 preoperatively to average 6.8 postoperatively (P < 0.001). Average maximum flow-rate (Q(max)) decreased from 6.3 ml/s preoperatively to 18.6 ml/ s postoperatively (P < 0.001). Preoperative average residual urine was 97 ml and reduced to average 9 ml after the operation. Temporary incontinence occurred in 4 cases, perioperative hemorrhage in 2, and urethral stricture in 1. CONCLUSION: Transurethral prostatectomy with the bipolar plasma kinetic technique is a safe and effective means for the treatment of BPH. PMID- 15755038 TI - [Diagnosis and treatment of urachal remnants with heterotopia sinus: a report of 2 cases]. AB - OBJECTIVE: To probe into the diagnosis and treatment of urachal remnants with heterotopic sinus. METHODS: Two cases with penis dorsal drainage for 5 approximately 6 years were diagnosed as urachal anomaly by sinus probing, methylene blue injection test, cystoscope examination, and type B ultrasonic examination, and both received surgical removal of the urachus and all the abnormal tissues associated with it. RESULTS: Surgery and pathology proved urachal cyst with communication drainage in both of the patients. Follow-up survey showed no recurrence and malignancy. CONCLUSION: Sinus probing, mythylene blue injection test and type B ultrasonic examination are effective methods for the diagnosis of urachal remnants with heterotopic sinus. Surgical treatment should include radical removal of the urachus and the abnormal tissues associated with it. PMID- 15755039 TI - [Chronic orchialgia]. AB - Chronic orchialgia, whose positive signs defy routine examinations, is usually misdiagnosised as chronic prostatitis or a psychological disorder. This paper aims at introducing the definition, etiology, diagnosis and management of chronic orchialgia, as well as a reappraisal and refinement of the traditional treatment of the disease. PMID- 15755040 TI - [Intracytoplasmic sperm injection for Klinefelter patients and the risk of chromosome anomaly in the patients' offspring]. AB - Patients with Klinefelter's syndrome are generally characterized by a 47, XXY karyotype, seminiferous tubule dysgenesis, azoospermia and infertility. However, focal spermatogenesis and severe oligozoospermia have been found in a few cases of 47, XXY, too. With the recent development in assisted reproductive technologies, the recovered spermatozoa by testicular biopsy from Klinefelter patients have been used for intracytoplasmic sperm injection (ICSI) and over 30 healthy neonates have been born. The conception of one 47, XXY fetus was found and then underwent abortion. This review focuses on the ICSI treatment of infertility in Klinefelter patients and the risk of chromosome anomaly in the offspring of these patients. PMID- 15755041 TI - [Effect of Zhenyangjian on the penile hemodynamics of the patients with arterial erectile dysfunction]. AB - OBJECTIVE: To explore an efficacious therapy for arterial erectile dysfunction (ED) by observing the effect of the Chinese traditional medicine Zhenyangjian on the penile hemodynamics of the patient. METHODS: Forty-seven patients with arterial ED, aged from 40 to 49 years, were treated with Zhenyangjian for 1 month. Intracavernous injection of papaverine and phentolamine (30 mg/l mg) was given and multifunctional color Doppler ultrasound was used to measure the peak systolic velocity(PSV) of the corpus cavernous arteries before and after Zhenyangjian administration. RESULTS: The PSV of the penis corpus cavernous arteries was obviously ameliorated in all the patients (P < 0.05). CONCLUSION: The Chinese traditional medicine Zhenyangjian can promote penile erection effectively. PMID- 15755042 TI - Tension-free polypropylene mesh for vaginal repair of anterior vaginal wall prolapse. AB - OBJECTIVE: To study the ongoing results of the repair of anterior vaginal wall prolapse reinforced with tension-free polypropylene mesh (GyneMesh, Gynecare, Ethicon, Issy-Les-Moulineaux, France). STUDY DESIGN: A case series of 87 consecutive women with anterior vaginal wall prolapse who underwent a transvaginal procedure using polypropylene mesh between October 1999 and August 2002. The mean age (+/-SD) was 62.4+/-13.4 years. Before the operation, patients underwent physical examination staging of the prolapse with the International Pelvic Organ Prolapse staging system. Thirteen women had stage 2 anterior vaginal wall prolapse (14.9%), 59 had stage 3 (67.9%), and 15 had stage 4 (17.3%). The polypropylene mesh was placed from the retropubic space to the inferior part of the bladder in a tension-freefashion. Patients were followed for 9-43 months, with a median follow-up (+/-SD) of 24+/-9.6 months. We defined "cure" as satisfactory (stage 1) or optimal (stage 0) outcome for point Ba in the staging system. RESULTS: Eighty-four patients returned for follow-up (96.6%). At follow up, 77 women were cured (91.6%), 5 women had asymptomatic stage 2 anterior vaginal wall prolapse, and 2 had a recurrent stage 3 (2.4%). There were no postoperative infections. There were a total of 7 vaginal erosions of the mesh (8.3%); 4 necessitated a second procedure for partial excision of the mesh. CONCLUSION: Vaginal repair of anterior vaginal wall prolapse reinforced with tension-free polypropylene mesh is effective and relatively safe. Vaginal erosion occurred in 8.3% of the study population but was easily manageable, with no sequelae. PMID- 15755043 TI - Magnetic resonance imaging of uterine version in a multiethnic, nulliparous, healthy female population. AB - OBJECTIVE: To determine measurement and topography of uterine position in asymptomatic women with different ethnicity. STUDY DESIGN: The angle and direction of uterine version were measured using magnetic resonance imaging in nulliparous, young volunteers from 5 ethnic groups (Emiratis, other Arabs, Indians/Pakistanis, Filipinos and Europeans/ Caucasians; N=55) and compared using Europeans/Caucasians as the reference group. RESULTS: The uterus was anteverted on the vagina in 46 (83.6%) and retroverted in 9 (16.4%) women, with no significant difference between Europeans/Caucasians and non-Caucasians. The angle of uterine version was significantly less (i.e., the cervix was more anteverted or retroverted on the vagina) in Europeans/Caucasians as compared to other women (p=0.002), particularly Indians/Pakistanis (p < 0.00001). CONCLUSION: The degree of uterine version is different in healthy, nulliparous European/Caucasian and non-Caucasian women. PMID- 15755045 TI - Contributions of physical and sexual abuse to women's experiences with chronic pelvic pain. AB - OBJECTIVE: To examine the roles of physical and sexual abuse in women with chronic pelvic pain using multi-dimensional pain assessment and to compare the chronic pelvic pain experiences of women with physical abuse to those of women with sexual abuse. STUDY DESIGN: Structured questionnaires were used to measure self-reported abuse, pain severity, psychological distress, physical functioning, interpersonalfunctioning, and coping in 63 women attending a tertiary care gynecologic clinic for diagnosis and treatment of chronic pelvic pain. RESULTS: Women with chronic pelvic pain who reported abuse demonstrated significantly more psychological distress than did women who reported no abuse, but there were no differences in pain severity, physical functioning, interpersonal functioning or coping. Women with physical abuse reported more overall psychological distress, depression, anxiety and somatization than women who reported no physical abuse. Women who reported sexual abuse showed more overall psychological distress and anxiety than women who reported no sexual abuse. While physical abuse was more consistently associated with psychological distress than was sexual abuse, both types of abuse were risk factors for distress. CONCLUSION: These results suggest that both physical and sexual abuse are associated with psychological distress in women with chronic pelvic pain but not with other domains of pain experience. Additional research to improve identification and treatment of women with both chronic pelvic pain and abuse is indicated. PMID- 15755044 TI - Use of a GnRH antagonist in controlled ovarian hyperstimulation for assisted conception in women with polycystic ovary disease: a randomized, prospective, pilot study. AB - OBJECTIVE: To compare the outcome of using gonadotropin-releasing hormone (GnRH) antagonists versus agonists in women with polycystic ovary disease (PCOD) who underwent controlled ovarian hyperstimulation (COH) for assisted reproductive techniques (ART). STUDY DESIGN: A total of 129 patients with PCOD were randomly allocated to undergo COH with a GnRH antagonist (59 patients) and GnRH agonist (leuprolide acetate) (70 patients) to prevent a premature luteinizing hormone (LH) surge. Assisted fertilization following oocyte retrieval and embryo transfer was performed. RESULTS: None of the cycles were cancelled due to a premature LH surge. There was no significant difference between the antagonist and agonist arms in the number of gonadotropin ampules consumed per cycle. However, in the antagonist arm a shorter duration of ovarian stimulation was recorded as compared to the agonist arm. Although similar numbers of oocytes was retrieved from both groups of patients, the quality of the oocytes, as measured by metaphase 2/total oocyte ratio, was lower in the antagonist arm as compared to the agonist arm. Pregnancy rates were 57.6% and 58.5% in the antagonist and agonist arms, respectively (p > 0.05). Implantation rates were not different (34.0% and 34.6%, respectively). The frequency of ovarian hyperstimulation syndrome also did not differ between the treatment groups (5% and 7.1%, respectively). CONCLUSION: The size of our study, on a specific subgroup of patients, does not allow a reliable conclusion regarding ART outcomefollowing the use of a GnRH antagonist versus agonist. Nevertheless, the protocol with the antagonist gave results that were as good as those of the protocol with the agonist in this PCOD patient population. PMID- 15755046 TI - One-stop hysteroscopy clinic for postmenopausal bleeding. AB - OBJECTIVE: To evaluate the role and feasibility of the "1-stop" clinic for management of postmenopausal bleeding, based on the use of transvaginal ultrasound and hysteroscopic examination under local anesthesia. STUDY DESIGN: A retrospective analysis of 308 patients referred to the clinic between October 2000 and May 2002 was carried out. Endometrial thickness of > 4 mm, as measured by transvaginal scan, was considered the cutoff limit for further investigation by hysteroscopy. RESULTS: Three hundred eight patients were seen in the 1-stop clinic. One hundred ten (35.7%) had endometrial thickness < or = 4 mm with no other associated abnormal findings and were discharged without proceeding to hysteroscopy. In 8 patients the scan was not conclusive, and they were offered hysteroscopy. Another 4 had a fluid-filled uterine cavity on sonography and were offered dilatation and curettage under general anesthesia. Eight patients opted to have the hysteroscopy done under general anesthesia, and in 12, hysteroscopyfailed under local anesthesia. Outpatient hysteroscopy was performed in 174 cases; 86 (49.4%) had normalfindings and were discharged. Endometrial polyps were found in 55 (31.6%) patients; in 42 the polyp was successfully removed at the same setting. Twenty patients (11.5%) had submucous fibroids and were discharged. A suspicious lesion was found in 13 (7.5%) patients; histologic examination confirmed endometrial carcinoma. A total of 216 of 308 (70%) patients were totally managed and discharged at the first visit with no further follow-up, and hospital admission was avoided in 258 (83.7%) of cases. CONCLUSION: The 1 stop clinic is effective in reducing the number of hospital visits per patient as well as hospital admissions and the waiting list. PMID- 15755047 TI - Breech presentation: evolution of management. AB - From the historical perspective, vaginal delivery of the persistent breech presentation had been the tradition since the 1st century A.D. The external cephalic version was perfected and popularized in the mid-16th century. A variety of instruments and maneuvers had been used in the 19th century, with successful application of forceps to the after-coming head, and in 1924, Edmund Piper developed forceps for application only to the after-coming head. Vaginal delivery of breech cases had been performed primarily for the safety of the mother. As blood banking, antibiotics and safe anesthesia became available and legal and the ethical and social milieu was changing, cesarean breech deliveries were performed more liberally to reduce the increased perinatal morbidity and mortality of infants born of vaginal breech deliveries. Liberalization of cesarean delivery resulted in increased maternal mortality, and increased maternal morbidity created potential hazards in subsequent pregnancies. Renewed interest in revival of the practice of external cephalic version created successful reduction in term breech presentation. Several retrospective, prospective and randomized studies of vaginal deliveries of some types of breech cases were conducted under strict, selective protocols, with results of outcome comparable to those of cesarean sections. The International Term Breech Collaborative Group, after study of a large number of frank and complete breech cases, concluded that planned cesarean delivery is a "substantially better method of delivery for the fetus" and that a policy of planned vaginal birth for term, singleton, breech fetuses should no longer be encouraged. PMID- 15755048 TI - Operative laparoscopy in tubal pregnancy: preoperative ultrasonographic measurement of hemoperitoneum as a predictor of blood loss and outcome. AB - OBJECTIVE: To study the relationship of ultrasonographic measurement of hemoperitoneum to estimated blood loss and outcome of operative laparoscopy in ectopic pregnancy. STUDY DESIGN: Retrospective study of all tubal pregnancy patients with preoperative ultrasonographic evidence of hemoperitoneum who underwent laparoscopy as part of the procedure at a tertiary referral center over a 3.5-year period. The largest transabdominal and transvaginal measurements of pelvic fluid collections in anterior-posterior, transverse and cranial-caudal dimensions were measured from all available preoperative ultrasound films. Patient characteristics, volume of estimated blood loss and complications were reviewed. RESULTS: A total of 29 patients were included in the study. Twenty-one underwent successful laparoscopic treatment (group 1). Four patients underwent conversion from laparoscopy to laparotomy because of bleeding (group 2). Four patients underwent conversion because of adhesions (group 3). There were no significant differences in patient characteristics. The single largest ultrasonographc diameter of pelvic fluid collection in any dimension seemed to correlate with estimated blood loss volume (R2 =.659; p <0.05), with significant volume present in those with a maximum diameter > 7 cm. The mean diameter of greatest measurements was significantly different between groups 1 and 2 and between groups 2 and 3 (p<0.05). However, such measurements did not predict the success of laparosocpic surgery. CONCLUSION: The greatest ultrasonographic measurement of pelvic fluid collection > or = 7 cm in any dimension appears to serve as a marker of significant hemoperitoneum. Hemoperitoneum alone does not appear to predict the outcome of surgery, and otherfactors should be considered prior to performing operative laparoscopy in ectopic pregnancy. PMID- 15755049 TI - Vulvoperineal reconstruction with a sulcus gluteus flap. AB - OBJECTIVE: To describe vulvoperineal reconstruction with a flap of the sulcus gluteus. STUDY DESIGN: Nonrandomized, prospective study in patients with vulvar cancer and vulvar intraepithelial neoplasia. After oncologic surgery, reconstruction with a sulcus gluteus flap was performed. The flap is supplied by the internal pudendal artery, and innervation is provided by the pudendal nerve. The tuber ischiale is the anatomic landmark for locating the pedicle. Follow-up was from 2 months to 2 years. Immediate and long-term complications were assessed. Aesthetic and functional results were analyzed and related to the quality of life and length of hospitalization. RESULTS: Vulvoperineal reconstruction was carried out in 6 patients who had undergone radical surgery. For reconstruction we used the sulcus gluteus flap in 4 unilateral cases and in 2 bilateral cases, for a total of 8 flaps. Cutaneous flaps of the sulcus gluteus provided good functional and aesthetic results. All flaps survived. There was no flap necrosis. The average length of hospitalization was 4 days. Complications were 2 breakdowns of the wound and 1 lymphocele. Postoperative discomfort was minimum. Patients were allowed to walk on day 6 and to sit on day 15. CONCLUSION: The advantages of this reconstruction are that it: is useful in cases of lymphadenectomy, is sensitive and maintains innervation, is distant enough from the receptor area to be used for large vulvar lesions, is able to hide the scar from the donor area in the sulcus gluteus, has a low complication rate, has a short hospitalization time and provides very good compliance with follow-up. PMID- 15755050 TI - Vulvar pain in women attending a general medical clinic in Accra, Ghana. AB - OBJECTIVE: To determine the prevalence of vulvar pain and pain related to sexual intercourse among women attending a general medical clinic in Accra, Ghana. STUDY DESIGN: A cross-sectional survey was performed. RESULTS: Forty-six women (22.8%) reported the presence of vulvar pain at the time of the study. There was significant association between the presence of vulvar pain and dissatisfaction with the quality of sexual activity (p < 0.001). There was no association between the presence of vulvar pain and the type of underwear used or the menstrual hygiene of the respondents. CONCLUSION: Vulvar pain with some relationship to sexual intercourse appears to be a significant problem among Ghanaian women. PMID- 15755051 TI - Erythropoietin use in a pregnant Jehovah's witness with anemia and beta thalassemia: a case report. AB - BACKGROUND: Anemia in pregnancy is seen often because of iron deficiency and the "physiologic dilution" that occurs in the third trimester. Other causes include genetic conditions, such as sickle cell anemia and thalassemias. In cases not responding to iron therapy, patients occasionally require a blood transfusion to restore adequate circulating red blood cell mass. In patients belonging to the Jehovah's Witness sect, transfusion of blood products is not allowed, and treatment of anemia in pregnancy may require use of erythropoietin. CASE: A 26 year-old, African American woman belonging to the Jehovah's Witness sect presented with anemia associated with beta-thalassemia. Iron therapy and prenatal vitamins did not correct the anemia, and the patient became symptomatic, with fatigue and shortness of breath when walking. Therapy with synthetic erythropoietin corrected the anemia, and the patient had an otherwise uncomplicated pregnancy and delivery. CONCLUSION: Synthetic erythropoietin has been used successfully in patients with renal failure and anemia. In pregnancy associated with renal failure and anemia, synthetic erythropoietin has been shown to be safe except for rare cases of hypertension. We treated anemia caused by beta-thalassemia in pregnancy with synthetic erythropoietin to avoid a transfusion in a Jehovah's Witness. PMID- 15755052 TI - 47,XXY female with testicular feminization and positive SRY: a case report. AB - BACKGROUND: Males with a 47,XXY karyotype have the clinical phenotype of Klinefelter syndrome. A few 47,XXY cases with a female phenotype have been reported. These individuals have positive SRY (testis-determining factor). The genetic explanation of this phenomenon is unclear. CASE: A 34-year-old woman presented with testicular feminization and a 47,XXY karyotype. Cytogenetic analysis and fluorescence in situ hybridization suggested that the Y chromosome had a normal structure; the polymerase chain reaction was positive for SRY. CONCLUSION: This is the third reported case of 47,XXY with afemale phenotype in spite of the presence of a Y chromosome and the normal SRY. This suggests that the phenotypic sex in these patients might be due to the involvement of other sex determining genes. PMID- 15755053 TI - Treating cornual pregnancy with a single methotrexate injection: a report of 3 cases. AB - BACKGROUND: Cornual pregnancy is a rare type of ectopic pregnancy. Methotrexate (MTX) has been used successfully for the treatment of tubal pregnancies; however, its use for interstitial pregnancies is less common. CASES: Three cases of cornual pregnancy were successfully treated with a single MTX injection (50 mg/m2), without complications. CONCLUSION: A single MTX injection as medical treatment is an alternative to surgical treatment of cornual pregnancies. PMID- 15755054 TI - [Final report on the 3rd convention of oncologists and radiologists of the CIS]. PMID- 15755055 TI - [Serum tumor marker TPS (tissue polypeptide-specific antigen)]. PMID- 15755056 TI - [Radiologic diagnosis of mediastinal lymphoma]. PMID- 15755057 TI - [Clinical evaluation of efficacy of chemoradiotherapy for Hodgkin's disease]. AB - Patients with stage III (A,B) Hodgkin's disease (366) received chemoradiotherapy consisting of 2-4 courses of combined modality treatment followed by total or subtotal irradiation of lymph nodes. Overall 10-year (84%) and 15-year (79%) and relapse-free 10-year (85%) and 15-year (82%) survival was reported in stage IIIA cases. Subtotal exposure proved relatively more effective in such patients without iliac and inguinal lymph node involvement. If, following combined modality therapy, intoxication symptoms were aborted in stage IIIB patients; fairly good results were obtained after total and subtotal irradiation of lymph nodes or involved areas (10-year (70%) and 15-year (65%) overall and 10-year (75%) and 15-year (75%) relapse-free survival. PMID- 15755058 TI - [Strontium-89 diagnosis and therapy of bone metastases]. AB - A clinical test of strontium-89 chloride effect was carried out in 150 patients with bone metastases and bone pain from different patterns of tumor. Different types of bone lesions were considered. Pain-relieving effect was reported in 77%, irrespective of lesion gravity. There was no response to strontium-89 chloride in 150 patients with bone metastases. It mostly occurred in cases of lysis. Strontium-89 chloride therapy contributed to partial bone tissue repair. It is indicated as a component of complex treatment for bone metastases. PMID- 15755060 TI - [Wilms tumor gene (WT1) expression in blood cells from patients with myelodysplastic syndrome]. PMID- 15755059 TI - [Prognosis of epidermoid anal carcinoma regression after conservative treatment]. AB - The prospective study was concerned with definition of the clinical and therapeutic factors behind poor response of anal cancer to radio- (RT) or chemoradiotherapy (CRT). Out of 64 female and 8 male patients at the mean age of 57 (33-81), thirty six had split-course of 60-65 Gy (RT), twenty--60-65 Gy, 5-FU and mitomycin C (CRT) and eighteen--up to 55-65 Gy (1.5 Gy--session 1, 1.0 Gy- session 2) (hyper-fractionated RT) plus 5-FU, for squamous cell anal carcinoma. There was no endorectal ultrasound evidence of perirectal lymph node involvement (uN0): T1-2uN-M0 (n=46), T3-4uN0M0 (n=11), uN1 or N2-3 (groin or endorectal ultrasound: T1-2uN-M0 (n=46), T3-4uN0M0 (n=11), uN1 or N2-3 (groin metastases) were detected in 7 patients: T1-2uN1-2M0 (n=7), T3-4N1-3M0 (n=10). Endorectal ultrasound staging (ERUS) used a linear 7.5 MHz transducer. The uTNM system was devised on the basis of tumor invasion parameters. There were no tumors confined to the subendothelial layer of the anal canal (uT1); 24 (32.4%) tumors were confined to the internal anal sphincter (uT2); 19 (25.7%) invaded the external anal sphincter (uT3) and 31 (41.9%)--levator ani (uT4). All carcinomas T4 (n=9) corresponded to the uT4 category. Only T-stage and tumor invasion (uT) proved significant prognostic variables. Complete response of T1-2 was 79.2%, T3-4- 33.3% (p=0.0003); uT2--95.8%, uT3--68.4%, and uT4--41.9% (except T4) (p=0.0001). In multivariate logistic analysis, uT alone appeared an independent variable (p=0.015). ERUS uTNM staging is more effective in prognosis for RT and CRT and, therefore, should be recommended for preliminary management of epidermoid anal carcinoma. PMID- 15755061 TI - [Ploidometrical evidence in support of the law of step-by-step development of cancer and its practical uses]. AB - The ploidometrical data on over 50,000 nuclei of intact and altered cells from cervical, endometrial, mammary, renal, pulmonary, thyroidal and cutaneous tissues at different stages of carcinogenesis showed theoretical and empirical evidence to match fully. That was interpreted as a mathematical support of the law of step by-step development of the disease. PMID- 15755062 TI - [The role of macrophages in asbestos-induced carcinogenesis]. AB - Data available on the pathways of asbestos (fibrous) carcinogenesis still leaves much to be desired. Asbestos is regarded as a non-genotoxic substance by most researchers. There is insufficient evidence on the interaction of fibres, target cells and macrophages. Macrophages secreted proteins (ca. 450 kD) to inhibit proliferation of intact mesothelium and cytoxine (3-5 kD) which stimulated the cellular sensitivity of intact mesothelium and mesotheliomas to the toxic influence of asbestos. It was suggested that the effect was due to the triggering of intrinsic causation of cell death. Like any other fibres, carcinogenic effect of asbestos could be accounted for by such significant factor as active oxygen radicals. When exposed to asbestos, both intact mesothelial and mesothelioma cells and macrophages synthesized those substances. Free radical-like substances in conjunction with macrophage-conditioned media produced toxic effect on mesothelial cells. The role of active oxygen radicals in fibre-induced carcinogenesis is discussed. PMID- 15755064 TI - [Ehrlich tumor cells stimulate T-cell production of interferon-gamma and are resistant to autocrine nitric oxide]. AB - Ehrlich tumor cells released factors which activated T-cell production of interferon-gamma and triggered a mechanism of nitric oxide (NO) production instead of interferon-gamma. When cultured in vitro, they produced NO without stimulation and responded to interferon-gamma by an output rate of NO significantly greater than that in bone marrow cells. Unlike P-815 cells, those of Ehrlich tumor proved resistant to autocrine NO. PMID- 15755063 TI - [Sodium nitrite-induced potentiation of spontaneous and 1,2-dimethylhydrazine induced carcinogenesis in male mice F1 (C57 B1xCBA)]. AB - Chronic sodium nitrite (SN) treatment potentiated spontaneous and 1,2 dimethylhydrazine (DMH)-induced carcinogenesis. Mice injected with SN alone showed a higher incidence of leukemia and lung cancer than in controls. Combined treatment with DMH and SN induced most of benign and malignant tumors (hepatic hemangioendothelioma, hepatocarcinoma, renal adenoma, etc.). The difference in the numbers of DMH- and SN-induced tumor bearers was not significant until a concentration of 500 mg/l was reached (64.7%). The level of multiple tumor incidence increased when SN 50 and 500 mg/l was used. Unlike DMH alone, cumulative incidence of DMH-specific tumors and leukemia after combined treatment was higher. An evaluation of cumulative incidence and relative risk established an indirect but positive correlation between SN dose, on the one hand, and spontaneous and induced carcinogenesis, on the other. The strongest carcinogenic effect was reported when DMH was used in combination with SN 500 mg/l. Our data confirmed the carcinogenic hazard of chronic exposure to SN which increased when in combination with that to a specific carcinogenic substance. PMID- 15755065 TI - [Investigation of allergenic properties of olipiphat]. AB - Olipiphat, which is experimentally capable of antitumor action, did not induce any immediate hypersensitivity, but lowered such response to horse serum; nor did it stimulate delayed hypersensitivity to full Freund adjuvant or sharp venous return after concavalin A. No skin or eye mucosa irritation was reported. PMID- 15755066 TI - [Role of cytologic and histologic examination in the diagnosis of thyroid diseases in patients exposed to radiation]. AB - Complex cytological and ultrasound study was conducted in 300 patients with thyroid pathology exposed to 5-250 cSv. Benign goiter changes and autoimmune thyroiditis were detected in 66.2% while malignant thyroidal nodules--in 3.7%. Benign and autoimmune changes alone were in 38%. Pathomorphological features of nodular changes and autoimmune thyroiditis patterns as well as their ultrasound representations were identified. PMID- 15755067 TI - [A comparison of two modalities of complex chemoradiotherapy for locally advanced nasopharyngeal carcinoma]. AB - Two modalities of complex chemoradiotherapy for locally advanced nasopharyngeal carcinoma stage III-IV were compared in the treatment of 52 patients. Thirty-four patients (group I) received polychemotherapy: cisplatin--100 mg/sq.m, i.v., dropwise, on day 4, plus ACOP (adriamycin 35 mg/sq.m, i.v.--4; cyclophosphamide 750 mg/sq.m, i.v.--4, day 1; vincristine 1.4 mg/sq.m, i.v.--4, and prednisolone 100 mg/sq.m, per os, days 1-5; group II (18)--cisplatin 100 mg/sq.m, day 4, plus 5-FU 1,000 mg, i.v.--4, injection, days 1-3. Full response was registered in 93.7% (group I) and 93.3% (group II). Overall 5-year and relapse-free survival was 85 and 63% (group I), and 83 and 56% (group II), respectively. There were no significant differences in radiation-related injury incidence in both groups. The best results were reported with chemoradiotherapy using two courses: an induction one and an adjuvant cisplatin drugs treatment concurrent with ACOP or 5-FU. PMID- 15755069 TI - [Methodology of bone marrow imaging in diagnosis of skeletal metastatic involvement in cancer patients]. AB - The cost-effectiveness of visualization techniques for the entire hemopoietic bone marrow (BM) in diagnosis of tumor involvement of the skeleton was evaluated in 1,200 cancer patients. BM involvement was diagnosed in 199. Generalized involvement detectable by imaging any skeletal region was reported in 72 (31.6%) patients. Focal BM involvement was registered in 127 (68.4%): pelvis--51, femur- 29, sternum--25, cervical or sternal region--25 and lumbar--15. When pelvis, sternum, femur and cervico-thoracic spine were imaged, tumors were detected in 90%, 3 regions--62.3-80.1%, 2--50.5-71.1% and 1--38.2-54.6%. PMID- 15755068 TI - [Rationale for surgical treatment of cancer of the lower pulmonary lobe with consideration for regional metastatic spread]. AB - Optimal extent of surgery for lung cancer is determined to a great degree by lymph drainage and regional metastases vis-a-vis tumor localization in the lobes. Data on surgical treatment of 505 cases of cancer of the lower lobe are presented. The study established such peculiarities of metastatic spread to mediastinal lymph collector as relatively frequent involvement of the lower mediastinal lymph nodes ("jumping" metastases included) and greater likelihood of spread to the superior mediastinal lymph nodes when those of radix pulmonis are involved. The latter should be interpreted as an indicator of lower lobe involvement. We established the significance of relatively worse prognosis for mediastinal lymph node involvement as well as high frequency of intrasternal recurrences of tumors located in the left lung. The prevailing evidence on partial resections of the right lung rather than lower bilobectomy and the good results of bilobectomies for stage T1-3N1-2 tumors suggest that conservative treatment might offer more advantage in managing cancer of the lower lobe of the right lung. Extended surgery entails lower incidence of intrasternal recurrence and is more effective when used to treat right-lung tumors. PMID- 15755070 TI - [Comparative clinical appraisal of the effectiveness and tolerability of paclitaxel-LANCE and taxol in combination chemotherapy for recurrences and platinum-refractory ovarian carcinoma]. AB - A clinical appraisal of efficacy and tolerability of taxol (T) and a generic drug -paclitaxel--LANCE (P)--was carried out in two groups of patients with recurrences (9) or platinum-refractory ovarian carcinoma (17). In group I, patients aged 19-71 (mean age--47 yrs) had received 6-7 courses, while, in group II, patients aged 44-72 (mean age--52 yrs) who suffered tumor progression--15.0 courses of various chemotherapeutic regimens. Following standard premedication, T and P were injected 135 mg/sq., intravenously, dropwise, during 3 hrs, as a rule, either concurrently with platinum drugs or with hemzar. In the final analysis, there was no clinically significant difference in overall response--44-30% and 44 48%, respectively, nor any significant difference in side-effect incidence was reported. PMID- 15755072 TI - [Therapy of cancer patients with concurrent critical atherosclerosis of the aorta and peripheral arteries]. AB - The results are presented of 5-year experience with therapy of concurrent neoplasia and atherosclerotic lesions of the aorta and peripheral arteries gained at the Center for Oncosurgery, Regional Oncological Dispensary, Chelyabinsk. The analysis was concerned with the treatment received by 118 patients, irrespective of tumor stage or localization: surgical correction of blood flow was carried out in 60. Clinically significant atherosclerotic lesions of the aorta and peripheral arteries in a cancer patient should not be regarded as absolute or relative contraindication for combined treatment. Atherosclerotic occlusion of the femoral or shin arteries was managed conservatively in most cases (p<0.05). In cases of similar lesions of the carotid arteries and aortal aneurysm, surgical correction of blood flow was mostly used. PMID- 15755071 TI - [Epidemiologic indices of cervical cancer morbidity in areas with various levels of industrial pollution]. AB - While general cervical carcinoma morbidity is on the decline in most of the surveyed Russian Federation areas characterized by varying levels of industrial pollution, the heavily industrialized region of Salavat revealed a different tendency. Its level was 1.6 times (p<0.05) those in other areas. That was due to the morbidity of women aged 40-69 and even 20-29 and 30-39 years. PMID- 15755073 TI - [Use of dexamethasone in treatment of high- and low-grade non-Hodgkin's lymphoma]. AB - Since it exerts a stronger antitumor action than predinisolone, dexamethasone was used for therapy of patients with non-Hodgkin's lymphoma refractory to CHOP. All patients (66), resistant to CHOP, suffered bone marrow involvement. Dexamethasone pulsed therapy was given to 45 patients, with 21 COP-BLAM or ASHAP, MAD, Dexa BEAM treated in control. Median response duration in high- and low-grade non Hodgkin's lymphoma groups was 2 and 12 months, respectively. PMID- 15755074 TI - [New results of endocrine therapy of breast cancer (the role of Aromasin)]. PMID- 15755075 TI - Characterization of antigen-specific CD8+ T lymphocyte responses in skin and peripheral blood following intradermal peptide vaccination. AB - Immune responses to cancer vaccines are commonly tested by measuring cutaneous reactions to intradermal (i.d.) antigen. When well-characterized peptide epitopes are injected i.d., infiltrates of CD4+ and CD8+ T lymphocytes are frequently seen. In this study, we have further characterized T cells derived from vaccine infiltrating lymphocyte (VIL) responses. We found that the infiltrates capable of producing IFN-gamma and cytolytic activity could recognize vaccine peptide, as well as antigen-positive melanoma cells. We studied antigen-specific T cell responses from VILs and peripheral blood in 10 patients who participated in a clinical trial. All patients received systemic Flt3 ligand (20 microg/kg/d) and i.d. peptides: Three NY-ESO-1 peptides, SLLMWITQCFL (157-167), SLLMWITQC (157 165), QLSLLMWIT (155-163); tyrosinase internal peptide YMDGTMSQV (368-376); Melan A/MART-1 analogue peptide ELAGIGILTV (26-35, E27L substitution); and influenza matrix peptide GILGFVFTL (58-66). In 54 paired VIL and peripheral blood analyses, a good correlation was found between responses in skin and in blood. These cells could be rapidly expanded in a short-term assay and thus appear to be memory T cells. The demonstrated presence of antigen-specific T cells at vaccination sites validates this method of assessing the immune response to i.d. vaccines. PMID- 15755076 TI - 2-Deoxystreptamine: central scaffold of aminoglycoside antibiotics. PMID- 15755077 TI - Over one hundred peptide-activated G protein-coupled receptors recognize ligands with turn structure. PMID- 15755078 TI - Alpha-fluorinated ethers, thioethers, and amines: anomerically biased species. PMID- 15755079 TI - BINOL: a versatile chiral reagent. PMID- 15755080 TI - Synthesis of beta-aminophosphonates and -phosphinates. PMID- 15755081 TI - Conjugate additions of nitroalkanes to electron-poor alkenes: recent results. PMID- 15755082 TI - Proteases universally recognize beta strands in their active sites. PMID- 15755083 TI - Concurrent tandem catalysis. PMID- 15755085 TI - Confined phonons in Si nanowires. AB - Raman microprobe studies of long crystalline Si nanowires reveal for the first time the evolution of phonon confinement with wire diameter. The Raman band at approximately 520 cm-1 in bulk Si is found to downshift and asymmetrically broaden to lower frequency with decreasing wire diameter D, in good agreement with a phenomenological model first proposed by Richter et al. An adjustable parameter (alpha) is added to the theory that defines the width of the Gaussian phonon confinement function. We find that this parameter is not sensitive to diameter over the range 4-25 nm. PMID- 15755086 TI - Metal recognition of septapeptides via polypod molecular architecture. AB - The understanding of the nature of recognition of inorganic materials by proteins is one of the core elements of and has profound implications in biological materials science and engineering. Using combinatorial display methods, a considerable number of short polypeptides have recently been selected with affinity to engineering materials. During these selections, more than several polypeptides are identified with binding specificity to a chosen inorganic material. Understanding the nature of surface recognition of materials by polypeptides is essential for rational design and biomimetic engineering of these inorganic-binding polypeptides for use as linkers, catalyzers, and growth modifiers in nanotechnology and nanobiotechnology. Although there may not be direct homology among the amino acids constituting the polypeptides, their function may come from conserved molecular architecture. Here we study crystallographic surface recognition of platinum metal-binding septapeptides by conformational analysis. We find that the septapeptides conform into certain molecular architectures containing multiple protrusions (polypods) that spatially match with the crystallographic metal surfaces. While the physical recognition may originate from how well the molecular polypods spatially match a given crystallographic surface, the degree of binding may be due to the reactive groups that form the polypods, e.g., charged or polar groups (e.g., hydroxyl and amine). These results are highly consistent with the experimental binding characteristics of the Pt binders with various degrees of affinities. PMID- 15755087 TI - Electronic signature of DNA nucleotides via transverse transport. AB - We report theoretical studies of charge transport in single-stranded DNA in the direction perpendicular to the backbone axis. We find that, if the electrodes which sandwich the DNA have the appropriate spatial width, each nucleotide carries a unique signature due to the different electronic and chemical structure of the four bases. This signature is independent of the nearest-neighbor nucleotides. Furthermore, except for the nucleotides with guanine and cytosine bases, we find that the difference in conductance of the nucleotides is large for most orientations of the bases with respect to the electrodes. By exploiting these differences it may be possible to sequence single-stranded DNA by scanning its length with conducting probes. PMID- 15755088 TI - Self-assembly of concentric quantum double rings. AB - We demonstrate the self-assembled formation of concentric quantum double rings with high uniformity and excellent rotational symmetry using the droplet epitaxy technique. Varying the growth process conditions can control each ring's size. Photoluminescence spectra emitted from an individual quantum ring complex show peculiar quantized levels that are specified by the carriers' orbital trajectories. PMID- 15755089 TI - Liquid crystalline nanowires in porous alumina: geometric confinement versus influence of pore walls. AB - Aligned liquid crystalline nanowires within ordered porous alumina templates show a pronounced texture on a macroscopic scale. We have investigated the influence of the geometric confinement and the nature of the pore walls on the mesophase formation by means of X-ray diffraction. The apparent texture is the result of a complex interplay of the pore geometry, interfacial phenomena, and the thermal history. Pores with a diameter of a few hundred nm guide the mesophase formation more efficiently than those with a diameter below 100 nm. PMID- 15755090 TI - Radial anisotropic growth of rhodium nanoparticles. AB - In this contribution, we report the synthesis of rhodium multipods that result from a homogeneous seeded growth mechanism. Small Rh nanocrystal seeds were synthesized by the reduction of RhCl3 in ethylene glycol in the presence of PVP. These seed particles could be subsequently used, without isolation, to form larger rhodium nanoparticles. A reaction temperature of 190 degrees C led to isotropic cubic Rh particles. Lowering the reaction temperature resulted in more anisotropic growth, which gave Rh cubes with horns at 140 degrees C, and Rh multipods at 90 degrees C. The anisotropic growth occurred in the (111) direction, as determined by high-resolution TEM (HRTEM). Anisotropic growth proceeds via a seeded growth mechanism, and not by oriented attachment. PMID- 15755091 TI - A "grow-in-place" architecture and methodology for electrochemical synthesis of conducting polymer nanoribbon device arrays. AB - Fully enclosed horizontal nanochannels, in a prearranged array on a substrate and with built-in electrical contacts and chemical access regions, were used as growth templates for electrochemical synthesis of conducting polymer nanoribbons. In this "grow-in-place" approach, the nanochannel templates are part of the final array structure and remain after fabrication of the nanoribbons. The built-in electrical contacts, which provide the electrical potential for electrochemical polymerization, also remain and become contacts/interconnects to the array components. The grow-in-place architecture and methodology remove the need for template dissolution, any post-synthesis nanoribbon "grow-and-then-place" manipulation, and any post-synthesis electrical contacting. The fact that the templates are fully enclosed prohibits dendrite formation during growth, ensures precise dimensionality, and gives the encapsulation needed in any real device application. In this report the grow-in-place approach to electrochemical polymerization is used to produce polyaniline nanoribbons. These were found to be fibrils and not tubes and to grow from the central region of the growth-template cross-section and not from the template walls. Two-point and four-point electrical characterization of these polyaniline nanoribbons, obtained using the built-in electrodes, was employed to yield the true polyaniline conductivity and to assess the ohmicity of the contacting approach. Conductivity studies, done as a function of nanoribbon width, show conductivity increases as the width decreases. We also show that our grow-in-place approach may be used for chemical polymerization. However, at least for polyaniline, electrochemical polymerization is superior since it does not suffer from diffusion-limited growth and allows precise placement of the nanoribbons in the growth channel. PMID- 15755092 TI - Selective growth of PbSe on one or both tips of colloidal semiconductor nanorods. AB - PbSe nanocrystals with rock-salt structure are grown on the tips of colloidal CdS and CdSe nanorods. The facets of wurtzite rods provide a substrate with various degrees of reactivity for the growth of PbSe. The presence of dangling Cd bonds may explain subtle differences between nonequivalent facets resulting in the selective nucleation of PbSe only on one of the two tips of each CdS rod. This approach has the potential to facilitate the fabrication of heterostructures with tailored optical and electronic properties. PMID- 15755093 TI - Chemical optimization of self-assembled carbon nanotube transistors. AB - We present the improvement of carbon nanotube field effects transistors (CNTFETs) performances by chemical tuning of the nanotube/substrate and nanotube/electrode interfaces. Our work is based on a method of selective placement of individual single walled carbon nanotubes (SWNTs) by patterned aminosilane monolayer and its use for the fabrication of self-assembled nanotube transistors. This method brings a relevant solution to the problem of systematic connection of self organized nanotubes. The aminosilane monolayer reactivity can be used to improve carrier injection and doping level of the SWNT. We show that the Schottky barrier height at the nanotube/metal interface can be diminished in a continuous fashion down to an almost ohmic contact through these chemical treatments. Moreover, sensitivity to 20 ppb of triethylamine is demonstrated for self-assembled CNTFETs, thus opening new prospects for gas sensors taking advantages of the chemical functionality of the aminosilane used for assembling the CNTFETs. PMID- 15755094 TI - Controlled growth of Si nanowire arrays for device integration. AB - Silicon nanowires were synthesized, in a controlled manner, for their practical integration into devices. Gold colloids were used for nanowire synthesis by the vapor-liquid-solid growth mechanism. Using SiCl4 as the precursor gas in a chemical vapor deposition system, nanowire arrays were grown vertically aligned with respect to the substrate. By manipulating the colloid deposition on the substrate, highly controlled growth of aligned silicon nanowires was achieved. Nanowire arrays were synthesized with narrow size distributions dictated by the seeding colloids and with average diameters down to 39 nm. The density of wire growth was successfully varied from approximately 0.1-1.8 wires/microm2. Patterned deposition of the colloids led to confinement of the vertical nanowire growth to selected regions. In addition, Si nanowires were grown directly into microchannels to demonstrate the flexibility of the deposition technique. By controlling various aspects of nanowire growth, these methods will enable their efficient and economical incorporation into devices. PMID- 15755095 TI - Precise and rapid size selection and targeted deposition of nanoparticle populations using CO2 gas expanded liquids. AB - This paper demonstrates the rapid and precise size selection of nanoparticle populations using the pressure tunable solvent properties of CO2-expanded liquids. Specifically, by pressurizing and expanding a single organic solution with carbon dioxide gas, ligand-stabilized silver particles of desired mean size were size selectively precipitated at desired locations. Compared to current techniques, this CO2-expanded liquid approach provides for faster and more efficient particle size separation, reduction in organic solvent usage, and pressure tunable size selection. PMID- 15755096 TI - Production and characterization of single-crystal FeCo nanowires inside carbon nanotubes. AB - We describe the synthesis of novel monocrystalline FeCo nanowires encapsulated inside multiwalled carbon nanotubes (MWNTs). These FeCo nanowires exhibit homogeneous Fe and Co concentrations and do not contain an external oxide layer due to the presence of insulating nanotube layers. The method involves the aerosol thermolysis of toluene-ferrocene-cobaltocene solutions in inert atmospheres. The materials have been carefully characterized using state-of-the art high-resolution transmission electron microscopy (HRTEM), electron-energy loss spectroscopy (EELS), scanning electron microscopy (SEM), energy-dispersive X ray analysis (EDX), electron diffraction, HREELS-STM elemental mapping, X-ray powder diffraction, and SQUID magnetometry. We noted that the formation of FeCo alloys occurs at relatively low pyrolytic temperatures (e.g., 650-750 degrees C). These single-crystal nanowires, which have not been reported hitherto, always exhibit the FeCo (110) plane parallel to the carbon nanotube axis. The FeCo nanomaterials have shown large coercive fields at room temperature (e.g., 900 Oe). We envisage that these aligned ferromagnetic nanowires could be used in the fabrication of high-density magnetic storage devices and magnetic composites. PMID- 15755097 TI - Gold nanocages: bioconjugation and their potential use as optical imaging contrast agents. AB - Gold nanocages of <40 nm in dimension have been synthesized using the galvanic replacement reaction between Ag nanocubes and HAuCl4 in an aqueous solution. By controlling the molar ratio between Ag and HAuCl4, the gold nanocages could be tuned to display surface plasmon resonance peaks around 800 nm, a wavelength commonly used in optical coherence tomography (OCT) imaging. OCT measurements on phantom samples indicate that these gold nanocages have a moderate scattering cross-section of approximately 8.10 x 10(-16) m2 but a very large absorption cross-section of approximately 7.26 x 10(-15) m2, suggesting their potential use as a new class of contrast agents for optical imaging. When bioconjugated with antibodies, the gold nanocages have also been demonstrated for specific targeting of breast cancer cells. PMID- 15755098 TI - Integrated semiconductor nanocrystal and epitaxical nanostructure systems: structural and optical behavior. AB - Integration of semiconductor epitaxical nanostructures and nanocrystals into two classes of quantum structures, uncovered adsorbed nanocrystals or buried via epitaxical overgrowth, is successfully demonstrated through structural and optical studies. The combination InGaAs/GaAs epitaxical structures and InAs nanocrystals is employed as a vehicle with the functional aim of exploiting the well developed optoelectronic communication technology based on the former with the biochemical and biomedical applications for which the latter are well suited. PMID- 15755099 TI - Hybrid epitaxial-colloidal semiconductor nanostructures. AB - We present a growth technique which combines wet-chemical growth and molecular beam epitaxy (MBE) to create complex semiconductor nanostructures with nanocrystals as active optical material. The obtained results show that wet chemically prepared semiconductor nanocrystals can be incorporated in an epitaxally grown crystalline cap layer. As an exemplary system we chose CdSe nanorods and CdSe(ZnS) core-shell nanocrystals in ZnSe and discuss the two limits of thin (d approximately 2R) and thick (d>2R) ZnSe cap layers of thickness d for CdSe nanorods and nanodots of radii R between 2 and 4 nm. In contrast to the strain-induced CdSe/ZnSe Stranski-Krastanow growth of a quantum dot layer in a semiconductor heterostructure, the technique proposed here does not rely on strain and thus results in additional degrees of freedom for choosing composition, concentration, shape, and size of the nanocrystals. Transmission electron microscopy and X-ray diffractometry show that the ZnSe cap layer is of high crystalline quality and provides all parameters for a consecutive growth of Bragg structures, waveguides, or diode structures for electrical injection. PMID- 15755100 TI - In-situ observation of switchable nanoscale topography for y-shaped binary brushes in fluids. AB - Direct, in-fluid observation of the surface morphology and nanomechanical properties of the mixed brushes composed of Y-shaped binary molecules PS-PAA revealed nanoscale network-like surface topography formed by coexisting stretched soluble PAA arms and collapsed insoluble PS chains in water. Placement of Y shaped brushes in different fluids resulted in dramatic reorganization ranging from soft repellent layer covered by swollen PS arms in toluene to an adhesive, mixed layer composed of coexisting swollen PAA and collapsed PS arms in water. These binary layers with the overall nanoscale thickness can serve as adaptive nanocoatings with stimuli-responsive properties. PMID- 15755101 TI - Chains, dimers, and sandwiches: melting behavior of DNA nanoassemblies. AB - We have studied, in experiment and theory, the melting behavior of DNA nanoassemblies of perylenediimide-bis-oligonucleotides (PON), in which two short oligonucleotide chains (16- or 24-mers) are attached to the extremities of a spacer molecule. By varying both the sequence orientation (exchange of 3' and 5' ends neighboring the spacer) and the oligomer composition of the solutions, nanoassemblies of different complexity can be generated. Our results show a subtle dependence of the melting behavior on the supramolecular arrangement of the DNA-based assemblies. PMID- 15755102 TI - A molecular switch based on potential-induced changes of oxidation state. AB - We have measured the conductance of a hepta-aniline oligomer attached to gold electrodes held under potential control in electrolyte. It increases fifteen-fold (to 5.3+/-0.4 nS) on oxidation from the leucoemeraldine form to the emeraldine salt. The single-molecule current-voltage characteristic, linear in toluene, displays negative differential resistance in an acidic electrolyte. The negative differential resistance is accounted for by modification of the local surface potential by the applied bias. These results connect electrochemical data directly to molecular electronic behavior in a two-terminal device. PMID- 15755103 TI - Photocurrent imaging of charge transport barriers in carbon nanotube devices. AB - The realization of high-performance electrical devices incorporating single-wall carbon nanotubes critically depends on the minimization of charge transport barriers in the tubes and at the contacts. Herein we demonstrate photocurrent imaging as a fast and effective tool to locate such barriers within individual metallic nanotubes contacted by metal electrodes. The locally induced photocurrents directly reflect the existence of built-in electric fields associated with the presence of depletion layers at the contacts or structural defects along the tubes. PMID- 15755104 TI - Spectroscopy of single- and double-wall carbon nanotubes in different environments. AB - Individual single-wall carbon nanotubes (SWNTs) and double-wall carbon nanotubes (DWNTs) were suspended in water for optical studies using sodium-cholate and other surfactants. We used time-resolved photoluminescence (PL) spectroscopy to study the influence of tube chirality and diameter as well as of the environment on nonradiative decay in small diameter tubes. The studies provide evidence for PL from small diameter core tubes in DWNTs and for a correlation of nonradiative decay with tube diameter and exciton red shift as induced by interaction with the environment. PMID- 15755105 TI - Observation of intrinsic size effects in the optical response of individual gold nanoparticles. AB - The photothermal heterodyne imaging method is used to study for the first time the absorption spectra of individual gold nanoparticles with diameters down to 5 nm. Intrinsic size effects that result in a broadening of the surface plasmon resonance are unambiguously observed. Dispersions in the peak energies and homogeneous widths of the single-particle resonances are revealed. The experimental results are analyzed within the frame of Mie theory. PMID- 15755106 TI - Labeling ribonuclease S with a 3 nm Au nanoparticle by two-step assembly. AB - We label ribonuclease S with a 3 nm Au nanoparticle (NP) by utilizing its two piece structure. One portion, S-peptide, is mutated with a unique NP attachment site. NP-peptide self-assembles with the other portion, S-protein, to form an active enzyme. NP mobility decreases with peptide labeling and S-protein association. Surface plasmon shifts support conjugation. Higher S-peptide coverages on the NP surface reduce nonspecific adsorption, while sterically hindering assembly of RNaseS. Thiols displace nonspecific adsorption, maximizing site-specific labeling. PMID- 15755107 TI - Modeling a suspended nanotube oscillator. AB - We present a general study of oscillations in suspended one-dimensional elastic systems clamped at each end, exploring a wide range of slack (excess length) and downward external forces. Our results apply directly to recent experiments in nanotube and silicon nanowire oscillators. We find the behavior to simplify in three well-defined regimes which we present in a dimensionless phase diagram. The frequencies of vibration of such systems are found to be extremely sensitive to slack. PMID- 15755108 TI - Electrostatic force-assisted nanoimprint lithography (EFAN). AB - We present and demonstrate a novel imprint method, electrostatic force-assisted nanoimprint lithography (EFAN), where a voltage applied between a mold and a substrate generates an electrostatic force that presses the mold into a resist on the substrate. We have successfully used EFAN to pattern nanostructures in a photocurable resist spin-coated on a wafer, with high fidelity and excellent uniformity over the entire substrate, in ambient atmosphere without using a vacuum chamber. In initial tests without any process optimization, 100 nm half pitch gratings with a residual layer thickness of 22+/-5 nm were imprinted across a 100 mm diameter wafer in about 2 s. Furthermore, numerical calculations show that the field magnitude experienced by the dielectric layers on the substrate is much less than their breakdown limit. Hence, EFAN is well suited for step-and repeat nanoimprint lithography, and its simple operation can simplify and speed up multilayer alignment process. PMID- 15755110 TI - Synthesis of blue luminescent si nanoparticles using atmospheric-pressure microdischarges. AB - Silicon nanoparticles are synthesized from a mixture of argon/silane in a continuous flow atmospheric-pressure microdischarge reactor. Particles nucleate and grow to a few nanometers (1-3 nm) in diameter before their growth is abruptly terminated in the short residence time microreactor. Narrow size distributions are obtained as inferred from size classification and imaging. As-grown Si nanoparticles collected in solution exhibit room-temperature photoluminescence that peaks at 420 nm with a quantum efficiency of 30%; the emission is stable for months in ambient air. PMID- 15755109 TI - Thermal transport in au-core polymer-shell nanoparticles. AB - Thermal transport in aqueous suspensions of Au-core polymer-shell nanoparticles is investigated by time-resolved measurements of optical absorption. The addition of an organic cosolvent to the suspension causes the polystyrene component of the polymer shell to swell, and this change in the microstructure of the shell increases the effective thermal conductivity of the shell by a factor of approximately 2. The corresponding time scale for the cooling of the nanoparticle decreases from 200 ps to approximately 100 ps. The threshold concentration of cosolvent that creates the changes in thermal conductivity, 5 vol % tetrahydrofuran in water or 40 vol % N,N-dimethylformamide in water, is identical to the threshold concentrations for producing small shifts in the frequency of the plasmon resonance. Because the maximum fraction of solvent in the polymer shell is less than 20 vol %, the increase in the effective thermal conductivity of the shell cannot be easily explained by contributions to heat transport by the solvent or enhanced alignment of the polystyrene backbone along the radial direction. PMID- 15755111 TI - An organometallic synthesis of TiO2 nanoparticles. AB - We report the synthesis of TiO2 nanoparticles that uses the low-temperature reaction of low-valent organometallic precursors. Bis(cyclooctatetraene)titanium reacts with dimethyl sulfoxide in organic solution at temperatures as low as room temperature to produce TiO2. In the absence of any supporting ligand, the reaction gives precipitation of amorphous TiO2 powder; however, in the presence of basic ligands such as tributylphosphine, tributylphosphine oxide and trioctylphosphine oxide, the precipitation is arrested, and chemically distinct, isolated, internally crystalline TiO2 nanoparticles are formed. PMID- 15755112 TI - Sculpting nanoelectrodes with a transmission electron beam for electrical and geometrical characterization of nanoparticles. AB - A method to produce metal electrodes with a gap of a few nanometers with a highly focused electron beam in a transmission electron microscope (TEM) is described. With this method the electrical and geometrical characterization of the same particle is possible. The I-V characteristics of a gold particle trapped between such electrodes showed the expected single-electron tunneling behavior, with a Coulomb gap corresponding to the geometry of the particle as observed with high resolution TEM. PMID- 15755113 TI - Charge transfer induced polarity switching in carbon nanotube transistors. AB - We probed the charge transfer interaction between the amine-containing molecules hydrazine, polyaniline, and aminobutyl phosphonic acid and carbon nanotube field effect transistors (CNTFETs). We successfully converted p-type CNTFETs to n-type and drastically improved the device performance in both the ON- and OFF transistor states, utilizing hydrazine as dopant. We effectively switched the transistor polarity between p- and n- type by accessing different oxidation states of polyaniline. We also demonstrated the flexibility of modulating the threshold voltage (Vth) of a CNTFET by engineering various charge-accepting and donating groups in the same molecule. PMID- 15755114 TI - Hydrogen transfer reaction on the surface of an oxide catalyst. AB - RuO2(110) exposes two kinds of active surface species (acidic and basic centers) that govern the interaction of the gas phase in contact with the catalyst's surface. Here we will elucidate the cooperative interplay of these two active surface sites for a simple model reaction, namely the water formation over RuO2 catalysts when supplying hydrogen and oxygen from the gas phase. The bridging O atoms harvest the hydrogen from the gas phase, while the on-top O atoms pick up those adsorbed hydrogen atoms from the bridging O atoms to form water. This mechanism of hydrogen transfer is mediated by a strong hydrogen bond. Hydrogen transfer is expected to play a vital role for the whole class of catalyzed hydrogenation and dehydrogenation reactions of hydrocarbons over RuO2. PMID- 15755115 TI - Remarkable supramolecular catalysis of glycoside hydrolysis by a cyclodextrin cyanohydrin. AB - (6AR,6DR)-6A,6D-di-C-cyano-beta-cyclodextrin (3) was synthesized and shown to catalyze hydrolysis of nitrophenyl glycosides with the reaction following Michaelis-Menten kinetics. At pH 7.4 and 25 degrees C, hydrolysis of 4 nitrophenyl-beta-glucopyranoside (2) was catalyzed with KM = 15 mM, kcat = 8.2 x 10-6 s-1, and kcat/kuncat = 1217. Catalysis was observed with concentration of 3 as low as 10 muM. Hydrolysis of the corresponding alpha-glucoside, alpha galactoside, alpha-mannoside, and 2-nitrophenyl-beta-galactoside was also catalyzed by 3, with kcat/kuncat ranging from 283 to 2147. A series of analogues of 3 was prepared and investigated for catalysis of the hydrolysis of 2: (6AR,6DR)-6A,6D-di-C-propyl-beta-cyclodextrin (9) was not catalytic, while 6A,6D di-C-cyano-6A,6D-dideoxy-beta-cyclodextrin (12) had a low catalytic activity (kcat/kuncat = 4). A kcat/kuncat = 48 was found for 6A,6D-dialdehydo-beta cyclodextrin dihydrate (11). It was proposed that 3 acts by general acid catalysis on the bound substrate. PMID- 15755116 TI - Enantioselective organocatalytic cyclopropanations. The identification of a new class of iminium catalyst based upon directed electrostatic activation. AB - A new method for enantioselective organocatalytic cyclopropanation is described. This study outlines the identification of a new class of iminium catalyst based on the concept of directed electrostatic activation (DEA). This novel organocatalytic mechanism exploits dual activation of ylide and enal substrates through a proposed electrostatic activation and stereodirected protocol. Formation of trisubstituted cyclopropanes with high levels of enantio- and diastereoinduction is accomplished for a variety of alpha,beta-unsaturated aldehydes and sulfonium ylides. In addition, mechanistic studies have found that this cyclopropanation reaction exhibits enantioselectivity and reactivity profiles that are in accord with the proposed DEA step. PMID- 15755117 TI - A simple organic reaction mediates the crystallization of the inorganic nanocluster [Ga13(mu3-OH)6(mu2-OH)18(H2O)24](NO3)15. AB - We report the single-crystal structure of an inorganic gallium cluster [Ga13(mu3 OH)6(mu2-OH)18(H2O)24](NO3)15.6H2O prepared using a simple organic reaction to drive the formation of the crystalline inorganic cluster. PMID- 15755118 TI - Ligand-catalyzed asymmetric alkynylboration of enones: a new paradigm for asymmetric synthesis using organoboranes. AB - Asymmetric conjugate alkynylation of enones may be effected using a B-1 alkynyldiisopropylboronate and catalytic amounts of binaphthol ligands. The high enantioselectivities observed are comparable to those obtained using stoichiometric amounts of binaphthol-modified boronate reagents, suggesting that this is a ligand-accelerated process. This is the first demonstration that catalytic amounts of chiral diols can be used with boronates to effect high levels of stereochemical control. PMID- 15755119 TI - Nonclassical crystallization: mesocrystals and morphology change of CaCO3 crystals in the presence of a polyelectrolyte additive. AB - Crystallization of calcite from differently concentrated calcium chloride solutions by the CO2 gas diffusion technique in the presence of polystyrene sulfonate yields crystal superstructures with unusual morphology. From the typical calcite rhombohedra as a starting situation, the morphology can be systematically varied via rounded edges and truncated triangles to finally concavely bended lens-like superstructures. Although these "crystals" are apparently well-faceted in light microscopy, electron microscopy analysis and BET reveal that the structures are highly porous and are composed of almost perfectly 3D-aligned calcite nanocrystals scaffolded to the final, partly nicely curved superstructures. At high supersaturations, superstructures with changed symmetry indicative of dipolar interaction potentials between the building blocks are found. The present model case also gives evidence for the importance of nonclassical, mesoscopic processes in crystallization in general. PMID- 15755120 TI - Rh(I)-catalyzed cyclization of 1-arylprop-2-yn-1-ol derivatives utilizing rhodium 1,4-migration. AB - A highly useful method for the construction of cyclopentanone derivatives fused with aromatic ring based on hydrorhodation-1,4-rhodium migration sequence is achieved. Treatment of 1-aryl-prop-2-yn-1-ols, which are easily accessible via an addition of acetylides to the corresponding aryl aldehydes, with a catalytic amount of [Rh(cod)2]2(BF4) and P(p-tolyl)3 in the presence of a base gives cyclopentanone derivatives in moderate to good yield. PMID- 15755121 TI - Differential inhibition of Staphylococcus aureus PBP2 by glycopeptide antibiotics. AB - The glycopeptide antibiotics prevent maturation of the bacterial cell wall by binding to the terminal d-alanyl-d-alanine moiety of peptidoglycan precursors, thereby inhibiting the enzymes involved in the final stages of peptidoglycan synthesis. However, there are significant differences in the biological activity of particular glycopeptide derivatives that are not related to their affinity for d-Ala-d-Ala. We compare the ability of vancomycin and a set of clinically relevant glycopeptides to inhibit Staphylococcus aureus PBP2 (penicillin binding protein), the major transglycosylase in a clinically relevant pathogen, S. aureus. We report experiments suggesting that activity differences between glycopeptides against this organism reflect a combination of substrate binding and secondary interactions with key enzymes involved in peptidoglycan synthesis. PMID- 15755122 TI - Synthesis of beta,beta-disubstituted vinyl boronates via the ruthenium-catalyzed Alder ene reaction of borylated alkynes and alkenes. AB - Ruthenium-catalyzed Alder ene reactions between borylated alkynes and terminal alkenes give the corresponding beta,beta-disubstituted vinyl boronates with high selectivity for the branched isomer. The stereochemistry of the vinyl boronate moiety was the result of a formal trans addition of the ene subunit across the alkyne, which is the opposite stereochemical outcome observed for other internal alkynes. PMID- 15755123 TI - In situ XAFS and NMR study of rhodium-catalyzed dehydrogenation of dimethylamine borane. AB - In situ X-ray absorption fine structure spectroscopy (XAFS) and 11B NMR were used to study the rhodium-catalyzed dimerization reaction of dimethylaminoborane, (CH3)2NHBH3 or DMAB. XAFS spectra show that the active form of the rhodium catalyst is most likely composed of a six-atom Rh core surrounded by tightly bound external ligands. NMR results show the presence of monomeric dimethylamine borane (CH3)2NBH2, providing evidence that hydrogen formation from the homogeneous Rh species occurs by an intramolecular pathway. This is in contrast to thermal pathways that involve intermolecular B-N concurrent with hydrogen formation. This work shows that in situ XAFS spectroscopy offers a unique experimental tool to differentiate between heterogeneous and homogeneous catalysis. PMID- 15755124 TI - Synthesis of the potent immunostimulatory adjuvant QS-21A. AB - QS-21A is one of the most promising new adjuvants for immune response potentiation and dose-sparing in vaccine therapy, given its exceedingly high level of potency and its favorable toxicity profile. Melanoma, breast cancer, small cell lung cancer, prostate cancer, HIV-1, and malaria are among the numerous maladies targeted in more than 80 recent and ongoing clinical trials involving QS-21A as a critical component for immune response augmentation in microgram doses. Herein is reported the first synthesis and structure verification of QS-21Aapi, applying novel glycosylation methodologies in the convergent modular construction of this rare and potent natural product immunostimulant. PMID- 15755125 TI - Nitrogen atom transfer between manganese complexes of salen, porphyrin, and corrole and characterization of a (nitrido)manganese(VI) corrole. AB - The investigations of complete nitrogen atom transfer reactions from (nitrido)manganese(V) salen to manganese(III) complexes of porphyrins and corroles revealed that stabilization of the [Mn(N)]2+ moiety is in the order of corrole > porphyrin > salen. The first kinetic examination of this quite fundamental reaction exposed a large solvent effect on both the enthalpy and entropy activation energies. Oxidation of the (nitrido)manganese(V) corroles leads to the first (nitrido)manganese(VI) complexes that are coordinated by tetrapyrrolic ligands. PMID- 15755126 TI - Single-crystalline and monodisperse LaF3 triangular nanoplates from a single source precursor. AB - Single-crystalline and monodisperse LaF3 triangular nanoplates (2.0 x 16.0 nm) in trigonal tysonite structure were synthesized by the thermolysis of a single source precursor (SSP), La(CF3COO)3, in a hot oleic acid/octadecene solution. The combined use of SSP and coordinating and noncoordinating solvents was demonstrated to have played key roles in the formation of such high-quality nanoplates, which could spontaneously organize into two types of superlattices (edge-to-edge and face-to-face) on a large area. This SSP approach has advantages of one-step, mass production, and easy operation, and may represent a rather general route toward metal fluoride nanocrystals. PMID- 15755127 TI - Crystal structure of BRL 42715, C6-(N1-methyl-1,2,3-triazolylmethylene)penem, in complex with Enterobacter cloacae 908R beta-lactamase: evidence for a stereoselective mechanism from docking studies. AB - BRL 42715, C6-(N1-methyl-1,2,3-triazolylmethylene)penem, is an active-site directed inactivator of bacterial beta-lactamases. The crystal structure of Enterobacter cloacae 908R class C beta-lactamase in complex with BRL 42715, docking, and energy minimization studies explain stereoselectivity of the binding of C6-(heterocyclic methylene)penems against class C beta-lactamase. PMID- 15755128 TI - Ordering of vacancies in type-I tin clathrate: superstructure of Rb8Sn44 square2. AB - One important step toward the understanding of the mechanisms of thermoelectric properties is the knowledge about local distortions and vacancy ordering in clathrates. The type-I clathrate Rb8Sn44 shows in single-crystal and powder diffraction patterns a 2 x 2 x 2 supercell of the primitive cubic unit cell (Pmn), which originates from an ordering of the partially occupied site. The latter is distributed around a 41 screw axis, and the vacancies lead to a relaxation of the tin framework, thereby creating a local distortion of the tetrakaidecahedron. PMID- 15755129 TI - Facile synthesis of silver nanoparticles useful for fabrication of high conductivity elements for printed electronics. AB - A facile synthesis of stable silver nanoparticles having a particle size of <10 nm is described. The synthesis involved reduction of silver acetate with a substituted hydrazine, such as PhNHNH2, in the presence of a 1-alkylamine, such as C16H33NH2, in toluene at 25-60 degrees C. Spin-coated thin films or printed electronic features of alkylamine-stabilized silver nanoparticles could be easily converted at 120-160 degrees C into highly conductive films or elements with conductivity of 2-4 x 104 S cm-1. Organic thin-film transistors with printed silver source/drain electrodes of this nature exhibited field-effect transistor properties which are similar to those of the devices using vacuum-deposited silver electrodes. PMID- 15755130 TI - Fabrication of colloidal crystals with tubular-like packings. AB - Methods are reported for the fabrication of colloidal crystal wires with tubular packings. Both free and silica-encased wires have been prepared. Porous silicon membranes are infiltrated with silica spheres, treated with silane, and annealed. After removal of the silicon template, short annealing times were found to result in colloidal crystal wires with varied packing geometries, while repeated annealing cycles produced a thin translucent silica sheath around the wires. Packing in the wires varies with the channel diameter of the Si membrane. The channels used in this study typically produce colloidal crystal wires with six strands, though wires with four to seven strands have been observed. Both chiral and achiral packings are also possible. PMID- 15755131 TI - Inhibition assay of biomolecules based on fluorescence resonance energy transfer (FRET) between quantum dots and gold nanoparticles. AB - An inhibition assay method was developed based on the modulation in the FRET efficiency between quantum dots (QDs) and gold nanoparticles (AuNPs) in the presence of the molecules which inhibit the interactions between QD- and AuNP conjugated biomolecules. For the functionalization, AuNPs were first stabilized by chemisorption of n-alkanethiols and then capped with the first generation polyamidoamine (G1 PAMAM) dendrimers. By employing a streptavidin-biotin couple as a model system, avidin was quantitatively analyzed as an inhibitor by sensing the change in photoluminescence (PL) quenching of SA-QDs by biotin-AuNPs. The detection limit for avidin was about 10 nM. It is anticipated that the PL quenching-based sensing system can be used for the quantitative analysis and high throughput screening of molecules which inhibit the specific biomolecular interactions. PMID- 15755132 TI - Atomic XAFS as a tool to probe the electronic properties of supported noble metal nanoclusters. AB - Atomic XAFS is a very attractive technique for probing electronic properties of supported metal nanoclusters. For platinum nanoparticles on different supports, the technique is found to be in good agreement with infrared CO adsorption measurements. The advantages of AXAFS, however, are that no probe molecule is required and that real-time measurements under reaction conditions are possible. PMID- 15755133 TI - Enantioselective synthesis of near enantiopure compound by asymmetric autocatalysis triggered by asymmetric photolysis with circularly polarized light. AB - Right- and left-handed circularly polarized light (CPL) has been proposed as one of the origins of homochirality of biomolecules. However, the enantiomeric excess induced by CPL has been only very low (<2% ee). We found the unprecedented example of asymmetric autocatalysis triggered directly by a chiral physical factor, that is, right- and left-handed CPL, leading to a near enantiopure compound. Asymmetric photolysis of racemic pyrimidyl alkanol by r-CPL irradiation followed by asymmetric autocatalysis affords (R)-pyrimidyl alkanol with >99.5% ee. On the other hand, irradiation with l-CPL affords (S)-pyrimidyl alkanol with >99.5% ee. Thus, chiral physical power, such as CPL, in conjunction with asymmetric autocatalysis, provides a highly enantioenriched compound. PMID- 15755134 TI - Enzymatic clipping of DNA wires coated with magnetic nanoparticles. AB - The study describes how DNA coated with magnetic nanoparticles remains biologically active and accessible to the BamH1 restriction enzyme. Long DNA molecules are coated with magnetic nanoparticles using electrostatic interactions. The coated, stretched, and surface-bound DNA is incubated in the restriction enzyme that specifically recognizes any strand containing the GGATCC base sequence and clips the DNA. We show that, despite the presence of the nanoparticles on the DNA, the enzyme is still able to recognize the cleavage site and effectively digest the assembly. PMID- 15755135 TI - A facile synthesis and properties of multicarbazole molecules containing multiple vinylene bridges. AB - Multibranched pi-conjugated materials and hyperbranched polymers have received considerable interest in both academic research and industrial applications because of their unusual molecular structures and properties. In this communication, we provide an efficient approach to synthesize two novel pi conjugated multibranch compounds, which have good solubility and are nanosize (about 5 nm). The double bonds of the multibranch compounds are synthesized using solvent-free Wittig-Horner-Emmons reactions. The reaction conditions are simple and mild. Their optical properties are discussed too. PMID- 15755136 TI - Label-free electronic detection of DNA-hybridization on nanogapped gold particle film. AB - A label-free high-resolution DNA sensing technique using a nanogapped gold particle film is presented to acquire nucleotide polymorphism information, which can be read by electronic protocols. PMID- 15755137 TI - [2]Borametallocenophanes of Zr and Hf: synthesis, structure, and polymerization activity. AB - The first [2]borametallocenophanes of Zr and Hf are reported, which were obtained from 1,2-bis(dimethylamino)-1,2-dicyclopentadienyldiborane(4). The constitution of these complexes in solution and, for the Hf complex, in the crystalline state is discussed. The title complexes were studied as Ziegler-Natta-type catalyst precursors for the polymerization of ethylene. Both complexes produced PE of high molecular mass up to 1.6 x 106 g/mol, and very interestingly, the Hf complex proved to be as active as its Zr analogue. PMID- 15755138 TI - Molecular salts with diquat-based electron acceptors for nonlinear optics. AB - We have used several techniques, including Stark spectroscopy and MO calculations, to investigate the optical and electronic properties of two new dipolar chromophoric cations containing diquat-based electron acceptor groups. Both the Stark data and the calculated parameters show that the strong electron accepting properties of a diquat unit lead to static first hyperpolarizabilities beta0 which are considerably larger than those of a related stilbazolium chromophore. In addition, one compound has a strongly 2D quadratic NLO response, providing a very rare example of a charged molecule displaying such behavior. PMID- 15755139 TI - Spectroscopic evidence for the supply of reactive oxygen during CO oxidation catalyzed by gold supported on nanocrystalline CeO2. AB - Nanocrystalline CeO2 supplies reactive oxygen in the form of surface eta1 superoxide species and peroxide adspecies at the one-electron defect site to the supported active species of gold for the oxidation of CO. PMID- 15755140 TI - Squaraine-derived rotaxanes: sterically protected fluorescent near-IR dyes. AB - A squaraine dye with bulky end groups is employed as the thread component in two Leigh-type amide rotaxanes. The rotaxanes are synthesized in a simple two-step process. X-ray crystal structures of the rotaxanes show that the pyridyl containing macrocycle is more rigid and wraps more tightly around the cyclobutene core of the squaraine thread compared to the isophthalamide-containing macrocycle. The rotaxanes exhibit photophysical properties that are similar to the precursor squaraine. The encapsulating macrocycle greatly increases the chemical stability of the squaraine thread and inhibits aggregation-induced broadening of its absorption spectrum. It should be possible to prepare squaraine derived rotaxanes with improved properties for a wide range of photophysical, photochemical, and biomedical applications. PMID- 15755141 TI - Imidazo[1,5-a]pyridine: a versatile architecture for stable N-heterocyclic carbenes. AB - The imidazo[1,5-a]pyridine skeleton provides a versatile platform for the generation of new types of stable N-heterocyclic carbenes. Rh(I) mono- (6) and biscarbenes (7) from imidazo[1,5-a]pyridin-3-ylidenes (ImPy) and derivatives such as 13 from a mesoionic carbene were synthesized and characterized. PMID- 15755142 TI - Evidence for radical fragmentations from persistent singlet carbenes. AB - Upon warm up, persistent aminohydrazinocarbenes undergo beta-fragmentation reactions. Experimental as well as computational evidences are reported for the involvement of radical processes. This is the first example of a homolytic fragmentation for a persistent singlet carbene. PMID- 15755143 TI - The normal range for secondary Swain-Schaad exponents without tunneling or kinetic complexity. AB - An analysis is presented of the range of secondary Swain-Schaad exponents to be expected at 25 degrees C in the absence of tunneling or kinetic complexity. From 15 996 sets of exact harmonic semiclassical equilibrium isotope effects for simple C-H/D/T exchange reactions and 954 sets of exact harmonic semiclassical secondary H/D/T kinetic isotope effects for C-H positions in simple organic reactions, the distribution of Swain-Schaad exponents versus magnitude of the isotope effect is determined. This distribution defines when a secondary Swain Schaad exponent may be considered to implicate nonsemiclassical behavior, revises the expected Swain-Schaad exponent for extrapolation of secondary isotope effects, and serves as a guide to the uncertainty in such extrapolations. PMID- 15755144 TI - A new synthetic method for alpha-alkoxycarbonyl iminium salt and its reaction with nucleophiles. AB - An iminium salt was easily prepared using the oxidation of amino ketene silyl acetal with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and the subsequent nucleophilic addition to this iminium species proceeded efficiently to afford alpha-amino esters in good yields. PMID- 15755145 TI - The role of Pd nanoparticles in ionic liquid in the Heck reaction. AB - Pd(0) nanoparticles with approximately 2 nm diameter, immobilized in 1-n-butyl-3 methylimidazolium hexafluorophosphate ionic liquid, are efficient catalyst precursors for coupling of aryl halides with n-butylacrylate. In situ TEM analysis of the ionic liquid catalytic solution after the catalytic reaction shows the formation of larger nanoparticles ( approximately 6 nm). The palladium content in the organic phase during the arylation reaction was checked by ICP-AS and shows significant metal leaching (up 34%) from the ionic phase to the organic phase at low substrate conversions and drops to 5-8% leaching at higher conversions. These results strongly suggest that the Pd(0) nanoparticles serve as a reservoir of "homogeneous" catalytic active species. PMID- 15755146 TI - Synthesis and fate of o-carboxybenzophenones in the biosynthesis of aflatoxin. AB - o-Carboxybenzophenones have long been postulated to be intermediates in the oxidative rearrangement of anthraquinone natural products to xanthones in vivo. Many of these Baeyer-Villiger-like cleavages are believed to be carried out by cytochrome P450 enzymes. In the biosynthesis of the fungal carcinogen, aflatoxin, six cytochromes P450 are encoded by the biosynthetic gene cluster. One of these, AflN, is known to be involved in the conversion of the anthraquinone versicolorin A (3) to the xanthone demethylsterigmatocystin (5) en route to the mycotoxin. An aryl deoxygenation, however, also takes place in this overall transformation and is proposed to be due to the requirement that an NADPH-dependent oxidoreductase, AflM, be active for this process to take place. What is known about other fungal anthraquinone --> xanthone conversions is reviewed, notably, the role of the o carboxybenzophenone sulochrin (25) in geodin (26) biosynthesis. On the basis of mutagenesis experiments in the aflatoxin pathway and these biochemical precedents, total syntheses of a tetrahydroxy-o-carboxybenzophenone bearing a fused tetrahydrobisfuran and its 15-deoxy homologue are described. The key steps of the syntheses entail rearrangement of a 1,2-disubstituted alkene bearing an electron-rich benzene ring under Kikuchi conditions to give the 2-aryl aldehyde 43 followed by silyltriflate closure to a differentially protected dihydrobenzofuran 44. Regiospecific bromination, conversion to the substituted benzoic acid, and condensation with an o-bromobenzyl alcohol gave esters 47 and 50. The latter could be rearranged with strong base, oxidized, and deprotected to the desired o-carboxybenzophenones. These potential biosynthetic intermediates were examined in whole-cell and ground-cell experiments for their ability to support aflatoxin formation in the blocked mutant DIS-1, defective in its ability to synthesize the first intermediate in the pathway, norsolorinic acid. Against expectation, neither of these compounds was converted into aflatoxin under conditions where the anthraquinones versicolorin A and B readily afforded aflatoxins B1 and B2. This outcome is evaluated further in a companion paper appearing later in this journal. PMID- 15755147 TI - Heme/non-heme diiron(II) complexes and O2, CO, and NO adducts as reduced and substrate-bound models for the active site of bacterial nitric oxide reductase. AB - As a first generation model for the reactive reduced active-site form of bacterial nitric oxide reductase, a heme/non-heme diiron(II) complex [(6L)Fe(II)...Fe(II)-(Cl)]+ (2) {where 6L = partially fluorinated tetraphenylporphyrin with a tethered tetradentate TMPA chelate; TMPA = tris(2 pyridyl)amine} was generated by reduction of the corresponding mu-oxo diferric compound [(6L)Fe(III)-O-Fe(III)-Cl]+ (1). Coordination chemistry models for reactions of reduced NOR with O2, CO, and NO were also developed. With O2 and CO, adducts are formed, [(6L)Fe(III)(O2-))(thf)...Fe(II)-Cl]B(C6F5)4 (2a x O2) {lambda(max) 418 (Soret), 536 nm; nu(O-O) = 1176 cm(-1), nu(Fe-O) = 574 cm(-1) and [(6L)Fe(II)(CO)(thf)Fe(II)-Cl]B(C6F5)4 (2a x CO) {nu(CO) 1969 cm(-1)}, respectively. Reaction of purified nitric oxide with 2 leads to the dinitrosyl complex [(6L)Fe(NO)Fe(NO)-Cl]B(C6F5)4 (2a x (NO)2) with nu(NO) absorptions at 1798 cm(-1) (non-heme Fe-NO) and 1689 cm(-1) (heme-NO). PMID- 15755148 TI - Analogues of vaccinia virus DNA topoisomerase I modified at the active site tyrosine. AB - The mechanism of type IB topoisomerase-mediated DNA relaxation was studied by modification of vaccinia topoisomerase I at the active site tyrosine (position 274) with several tyrosine analogues. These analogues had varied steric, electronic, and stereochemical features to permit assessment of those structural elements required to support topoisomerase function. Eleven tyrosine analogues were successfully incorporated into the active site of vaccinia topoisomerase I. It was found that only tyrosine analogues having the phenolic -OH group in the normal position relative to the protein backbone were active. Modifications that replaced the nucleophilic tyrosine OH (pKa approximately 10.0) group with NH2 (pKa 4.6), SH (pKa approximately 7.0), or I groups or that changed the orientation of the nucleophilic OH group essentially eliminated topoisomerase I function. For the active analogues, the electronic effects and H-bonding characteristics of substituents in the meta-position of the aromatic ring may be important in modulating topoisomerase I function. The pH profile for the functional analogues revealed a small shift toward lower pH when compared with wild-type topoisomerase I. PMID- 15755149 TI - A new four-base genetic helix, yDNA, composed of widened benzopyrimidine-purine pairs. AB - We describe the properties of stable DNA-like self-assembled helices composed entirely of base pairs involving two new size-expanded pyrimidines. We term this new helix geometry "yDNA" (an abbreviation of "wide DNA"). The new pyrimidine analogues, yT and yC, are increased in size by benzo-homologation and have a geometry that is distinct from previous size-expanded pyrimidines. The yT and yC deoxyribosides were incorporated into oligodeoxynucleotides designed to form four pairs: yT-A, A-yT, yC-G, and G-yC. Helices were characterized by thermal denaturation, mixing data, and circular dichroism spectra. Results showed that highly stable double-stranded helices were formed in several sequence contexts. The data further showed that yT and yC could be segregated onto one strand and used to bind to natural strands of DNA with high sequence selectivity. The combination of yC, yT, G, and A make up a new selective, self-assembling four base genetic pairing system that functions in many respects like natural DNA, but which is structurally distinct. The results establish that multiple variants of size-expanded DNA-like helices are feasible and suggest the possibility of a future eight-base genetic system based on the yDNA geometry. Finally, the high binding selectivity, affinity, and fluorescence of yDNA strands may yield useful applications in detection of nucleic acid sequences. PMID- 15755150 TI - Effect of PNA backbone modifications on cyanine dye binding to PNA-DNA duplexes investigated by optical spectroscopy and molecular dynamics simulations. AB - Optical spectroscopy and molecular dynamics simulations have been used to study the interaction between a cationic cyanine dye and peptide nucleic acid (PNA)-DNA duplexes. This recognition event is important because it leads to a visible color change, signaling successful hybridization of PNA with a complementary DNA strand. We previously proposed that the dye recognized the minor groove of the duplex, using it as a template for the assembly of a helical aggregate. Consistent with this, we now report that addition of isobutyl groups to the PNA backbone hinders aggregation of the dye when the substituents project into the minor groove but have a weaker effect if directed out of the groove. UV-Visible and circular dichroic spectroscopy were used to compare aggregation on the different PNA-DNA duplexes, while molecular dynamics simulations were used to confirm that the substituents block the minor groove to varying degrees, depending on the configuration of the starting amino acid. In addition to a simple steric blockage effect of the substituent, the simulations suggest that directing the isobutyl group into the minor groove causes the groove to narrow and the duplex to become more rigid, structural perturbations that are relevant to the growing interest in backbone-modified PNA for applications in the biological and materials sciences. PMID- 15755151 TI - Effect of proline and glycine residues on dynamics and barriers of loop formation in polypeptide chains. AB - Glycine and proline residues are frequently found in turn and loop structures of proteins and are believed to play an important role during chain compaction early in folding. We investigated their effect on the dynamics of intrachain loop formation in various unstructured polypeptide chains. Loop formation is significantly slower around trans prolyl peptide bonds and faster around glycine residues compared to any other amino acid. However, short loops are formed fastest around cis prolyl bonds with a time constant of 6 ns for end-to-end contact formation in a four-residue loop. Formation of short loops encounters activation energies in the range of 15 to 30 kJ/mol. The altered dynamics around glycine and trans prolyl bonds can be mainly ascribed to their effects on the activation energy. The fast dynamics around cis prolyl bonds, in contrast, originate in a higher Arrhenius pre-exponential factor, which compensates for an increased activation energy for loop formation compared to trans isomers. All atom simulations of proline-containing peptides indicate that the conformational space for cis prolyl isomers is largely restricted compared to trans isomers. This leads to decreased average end-to-end distances and to a smaller loss in conformational entropy upon loop formation in cis isomers. The results further show that glycine and proline residues only influence formation of short loops containing between 2 and 10 residues, which is the typical loop size in native proteins. Formation of larger loops is not affected by the presence of a single glycine or proline residue. PMID- 15755152 TI - Electrochemical and guest binding properties of Frechet- and Newkome-type dendrimers with a single viologen unit located at their apical positions. AB - Several new series of dendrimers containing a single redox-active 4,4' bipyridinium (viologen) residue were synthesized and characterized. In these dendrimers, the viologen group is covalently attached to the apical position of a Newkome- or Frechet-type dendron, ranging in size from first to third generation of growth. The half-wave potentials corresponding to the two consecutive one electron reductions of the viologen residue are affected by the size of the dendritic component. The size effects are more pronounced in the Newkome-type dendrimers and seem to result from the polarity contrast between the microenvironments provided by the solution and the internal phase of the dendrimer. Unlike in many other dendrimers having a redox-active core, the voltammetric behavior remains fast (reversible) even in third generation dendrimers. Pulse gradient stimulated echo NMR diffusion coefficient measurements on the Newkome-type dendrimers reveal that their hydrodynamic radii are relatively invariant in solvents of widely different polarities (dichloromethane to dimethyl sulfoxide). The host-guest binding interactions between the viologen residue in these dendrimers and the crown ether host bis-p-phenylene-34-crown-8 were also investigated. While in Newkome-type dendrimers the growth of the dendron caused a substantial attenuation of the binding constant values, this size effect was not observed in the Frechet-type dendrimers. These electrochemical and binding measurements underscore some of the structural differences between these two common types of dendritic architectures. PMID- 15755153 TI - A luminescent receptor with affinity for N-terminal histidine in peptides in aqueous solution. AB - Crown ethers of suitable size are the perfect artificial host compounds for ammonium ion binding, but the rather low affinity in aqueous solution prevents their use at physiological conditions. We report here the synthesis and properties of a luminescent benzo crown ether with a pendant copper imidodiacetic acid complex, which coordinates with high affinity to histidine. The emission intensity of the benzo crown ether increases significantly in the presence of ammonium ions in methanol. At physiological conditions in buffered water at pH 7.5 these interactions are too weak to be detected. If an ammonium ion and an imidazole moiety are present in the analyte, such as in His-Lys-OMe or His-OMe, high binding affinity in aqueous solution is restored. The binding event is signaled by an increase in emission intensity, which can even be observed with the naked eye. This allows the selective detection of small peptides containing N terminal histidine or histidine among all other amino acids at physiological conditions. PMID- 15755154 TI - Mineralization of synthetic polymer scaffolds: a bottom-up approach for the development of artificial bone. AB - The controlled integration of organic and inorganic components confers natural bone with superior mechanical properties. Bone biogenesis is thought to occur by templated mineralization of hard apatite crystals by an elastic protein scaffold, a process we sought to emulate with synthetic biomimetic hydrogel polymers. Cross linked polymethacrylamide and polymethacrylate hydrogels were functionalized with mineral-binding ligands and used to template the formation of hydroxyapatite. Strong adhesion between the organic and inorganic materials was achieved for hydrogels functionalized with either carboxylate or hydroxy ligands. The mineral nucleating potential of hydroxyl groups identified here broadens the design parameters for synthetic bonelike composites and suggests a potential role for hydroxylated collagen proteins in bone mineralization. PMID- 15755155 TI - A molecular flytrap for the selective binding of citrate and other tricarboxylates in water. AB - The synthesis and binding properties of a new tricationic guanidiniocarbonyl pyrrole receptor 7 are described. Receptor 7 binds citrate 9 and other tricarboxylates such as trimesic acid tricarboxylate 8 with unprecedented high association constants of K(assoc) > 10(5) M(-1) in water as determined by UV and fluorescence tritration studies. According to NOESY experiments and molecular modeling calculations, the tricarboxylates are bound within the inner cavity of receptor 7 by ion pairing between the carboxylate groups and the guanidiniocarbonyl pyrrole moieties, favored by the nonpolar microenvironment of the cavity. Hence, receptor 7 can be regarded as a molecular flytrap. In the case of the aromatic tricarboxylate 8, additional aromatic interactions further strengthen the complex. The complexes with the tricarboxylates are so strong that even the presence of a large excess of competing anions or buffer salts does not significantly affect the association constant. For example, the association constant for citrate changes only from K(assoc) = 1.6 x 10(5) M(-1) in pure water to K(assoc) = 8.6 x 10(4) M(-1) in the presence of a 170-fold excess of bis-tris buffer and a 1000-fold excess of chloride. This makes 7 one of the most efficient receptors for the binding of citrate in aqueous solvents reported thus far. PMID- 15755156 TI - Total synthesis and properties of the crambescidin core zwitterionic acid and crambescidin 359. AB - The total synthesis of the crambescidin core acid 9, crambescidins 359 (8) and 431 (7), and the properties of the crambescidin core are described. A key step of the synthetic route to guanidinium carboxylate 9 is Pd(0) catalyzed cleavage of the ester side chain of pentacyclic cinnamyl ester 15. This ester is also employed to prepare a small library of crambescidin alkaloid analogues that differ in their C14 side chain. The zwitterionic guanidinium carboxylate 9 was shown to readily decarboxylate to form crambescidin 359 (8). Decarboxylation of crambescidin core acid 9 was fastest under basic conditions. In the presence of base, up to eight deuterium atoms can be incorporated into the pentacyclic crambescidin core. Both deuterium incorporation and decarboxylation of crambescidin core acid 9 are the result of facile ring opening of the spirocyclic ether rings of the pentacyclic guanidinium moiety. PMID- 15755157 TI - Intramolecular electron transfer in bis(methylene) adamantyl radical cation: a case study of diabatic trapping. AB - Our characterization of the potential energy surface for electron transfer (ET) in the bis(methylene)adamantane (BMA) model radical cation shows that the surface topology is prone to diabatic trapping (competition between ET and upward hops to the excited state). The general conditions for this phenomenon have been derived. The surface is centered around a conical intersection, and diabatic trapping occurs because one of the branching space coordinates (coordinates that lift the degeneracy at first order) corresponds to a vector of small length. For BMA, this coordinate is an antisymmetric breathing mode of the rigid carbon framework. Other modes (including methylene torsions and pyramidalizations) may lift the degeneracy at second-order but do not affect the energy gap at the intersection region effectively. The resulting topology is similar to that of an (n - 1) dimensional seam (where n is the number of nuclear degrees of freedom of the molecule) that cannot be avoided along the reaction coordinate, thus favoring recrossing to the upper surface. This analysis is extended by ab initio semiclassical dynamics using an Ehrenfest and a trajectory surface hopping algorithm implemented at the CASSCF level. Examination of the trajectories shows that there is no single mode that controls the diabatic trap, in agreement with the condition that there is no predominant degeneracy-lifting coordinate. Thus the reactivity depends on a combination of small effects, where presumably higher order effects come into play. This should be the general behavior of dynamics at a diabatic trapping situation. PMID- 15755158 TI - Synthesis and application of poly(phenylene ethynylene)s for bioconjugation: a conjugated polymer-based fluorogenic probe for proteases. AB - A set of carboxylate-functionalized poly(phenylene ethynylene)s (PPEs) has been synthesized in which the carboxylic acid groups are separated from the polymer backbone by oligo(ethylene glycol) spacer units. These polymers are soluble in water and organic solvents and have photophysical properties that are sensitive to solvent conditions, with high salt content and the absence of surfactant promoting the formation of aggregates of relatively low quantum yield and long fluorescence lifetime. Quenching of these materials by the dinitrophenyl (DNP) chromophore (K(SV) approximately 10(4)) is also highly solvent-dependent. The presence of carboxylate groups far from the polymer backbone appended to each repeating unit allows for the postpolymerization modification of these PPEs with peptides by methods analogous to those described for carboxylate-functionalized small-molecule dyes. Covalent attachment of the fluorescence-quenching 14-mer Lys(DNP)-GPLGMRGLGGGGK to the PPE results in a nonemissive substrate whose fluorescence is restored upon treatment with trypsin. The rate of fluorescence turn-on in this case is increased 3-fold by the presence of surfactant, though the actual rate of peptide hydrolysis remains the same. A small-molecule mimic of the polymer-peptide system shows a smaller fluorescence enhancement upon treatment with trypsin, illustrating the value of polymer-based amplification in this sensory scheme. PMID- 15755159 TI - Ruthenium-catalyzed aromatization of enediynes via highly regioselective nucleophilic additions on a pi-alkyne functionality. A useful method for the synthesis of functionalized benzene derivatives. AB - TpRu(PPh3)(CH3CN)2 PF6 (10 mol %) catalyst effected the nucleophilic addition of water, alcohols, aniline, acetylacetone, pyrroles, and dimethyl malonate to unfunctionalized enediynes under suitable conditions (100 degrees C, 12-24 h) and gave functionalized benzene products in good yields. In this novel cyclization, nucleophiles very regioselectively attack the internal C1' alkyne carbon of enediynes to give benzene derivatives as a single regioisomer. Experiments with methoxy substituents exclude the possible involvement of naphthyl cations as reaction intermediates in the cyclization of (o-ethynylphenyl) alkynes. Deuterium labeling experiments indicate that the catalytically active species is ruthenium pi-alkyne rather than ruthenium-vinylidene species. This hypothesis is further confirmed by the aromatization of o-(2'-iodoethynyl)phenyl alkynes with alcohols. We propose a nucleophilic addition/insertion mechanism for this nucleophilic aromatization on the basis of a series of experiments. PMID- 15755160 TI - Catalytic asymmetric cyano-ethoxycarbonylation reaction of aldehydes using a YLi3 tris(binaphthoxide) (YLB) complex: mechanism and roles of achiral additives. AB - Full details of a catalytic asymmetric cyano-ethoxycarbonylation reaction promoted by a heterobimetallic YLi3 tris(binaphthoxide) complex (YLB 1), especially mechanistic studies, are described. In the cyanation reaction of aldehydes with ethyl cyanoformate, three achiral additives, H2O, tris(2,6 dimethoxyphenyl)phosphine oxide (3a), and BuLi, were required to achieve high reactivity and enantioselectivity (up to >99% yield and up to 98% ee). The roles of achiral additives and the reaction pathway were investigated in detail. In situ IR analysis revealed that the initiation step to generate LiCN from H2O, BuLi, and ethyl cyanoformate is rather slow. On the basis of mechanistic studies of the initiation step to generate an active nucleophilic species, reaction conditions were optimized by using a catalytic amount of acetone cyanohydrin as an initiator. Under the optimized conditions, the induction period decreased and the reaction completed within 9 min using 5 mol % YLB at -78 degrees C. Catalyst loading was successfully reduced to 1 mol %. Kinetic experiments and evaluation of the substituent effects of phosphine oxide revealed that phosphine oxide had beneficial effects on both the reaction rate and the enantioselectivity. The putative active species as well as the catalytic cycle of the reaction are also discussed. PMID- 15755161 TI - Ligand-assisted reduction of osmium tetroxide with molecular hydrogen via a [3+2] mechanism. AB - Osmium tetroxide is reduced by molecular hydrogen in the presence of ligands in both polar and nonpolar solvents. In CHCl3 containing pyridine (py) or 1,10 phenanthroline (phen), OsO4 is reduced by H2 to the known Os(VI) dimers L2Os(O)2(mu-O)2Os(O)2L2 (L2 = py2, phen). However, in the absence of ligands in CHCl3 and other nonpolar solvents, OsO4 is unreactive toward H2 over a week at ambient temperatures. In basic aqueous media, H2 reduces OsO4(OH)n(n-) (n = 0, 1, 2) to the isolable Os(VI) complex, OsO2(OH)4(2-), at rates close to that found in py/CHCl3. Depending on the pH, the aqueous reactions are exergonic by deltaG = 20 to -27 kcal mol(-1), based on electrochemical data. The second-order rate constants for the aqueous reactions are larger as the number of coordinated hydroxide ligands increases, k(OsO4) = 1.6(2) x 10(-2) M(-1) s(-1) < k(OsO4(OH)-) = 3.8(4) x 10(-2) M(-1) s(-1) < k(OsO4(OH)2(2-)) = 3.8(4) x 10(-1) M(-1) s(-1). The observation of primary deuterium kinetic isotope effects, k(H2)/k(D2) = 3.1(3) for OsO4 and 3.6(4) for OsO4(OH)-, indicates that the rate-determining step in each case involves H-H bond cleavage. Density functional calculations and thermochemical arguments favor a concerted [3+2] addition of H2 across two oxo groups of OsO4(L)n and argue against H* or H- abstraction from H2 or [2+2] addition of H2 across one Os=O bond. The [3+2] mechanism is analogous to that of alkene addition to OsO4(L)n to form diolates, for which acceleration by added ligands has been extensively documented. The observation that ligands also accelerate H2 addition to OsO4(L)n highlights the analogy between these two reactions. PMID- 15755162 TI - A metastable prerequisite for the growth of lumazine synthase crystals. AB - Dense liquid phases, metastable with respect to a solid phase, form in solutions of proteins and small-molecule materials. They have been shown to serve as a prerequisite for the nucleation of crystals and other ordered solid phases. Here, using crystals of the protein lumazine synthase from Bacillus subtilis, which grow by the generation and spreading of layers, we demonstrate that within a range of supersaturations the only mechanism of generation of growth layers involves the association of submicrometer-size droplets of the dense liquid to the crystal surface. The dense liquid is metastable not only with respect to the crystals, but also with respect to the low-concentration solution: dynamic light scattering reveals that the droplets' lifetime is limited to several seconds, after which they decay into the low-concentration solution. The short lifetime does not allow growth to detectable dimensions so that liquid-liquid phase separation is not observed within a range of conditions broader than the one used for crystallization. If during their lifetime the droplets encounter a crystal surface, they lower their free energy not by decay, but by transformation into crystalline matter, ensuring perfect registry with the substrate. These observations illustrate two novel features of phase transformations in solutions: the existence of doubly metastable, short-lifetime dense phases and their crucial role for the growth of an ordered solid phase. PMID- 15755163 TI - Comparison of analytical techniques for purity evaluation of single-walled carbon nanotubes. AB - We compare popular analytical techniques, including scanning and transmission electron microscopy (SEM and TEM), thermogravimetric analysis (TGA), and Raman and near-infrared (NIR) spectroscopy, for the evaluation of the purity of bulk quantities of single-walled carbon nanotubes (SWNTs). Despite their importance as imaging techniques, SEM and TEM are not capable of quantitatively evaluating the purity of typical inhomogeneous bulk SWNT samples because the image frame visualizes less than 1 pg of SWNT material; furthermore, there is no published algorithm to convert such images into numerical data. The TGA technique is capable of measuring the amount of metal catalyst in an SWNT sample, but does not provide an unambiguous separation between the content of SWNTs and carbonaceous impurities. We discuss the utilization of solution-phase near-infrared spectroscopy and solution-phase Raman spectroscopy to quantitatively compare arbitrary samples of bulk SWNT materials of different purities. The primary goal of this paper is to provide the chemical community with a realistic evaluation of current analytical tools for the purity evaluation of a bulk sample of SWNTs. The secondary goal is to draw attention to the growing crisis in the SWNT industry as a result of the lack of quality control and the misleading advertising by suppliers of this material. PMID- 15755164 TI - Mononuclear aluminum hydroxide for the design of well-defined homogeneous catalysts. AB - An unprecedented aluminum hydroxide LAlMe(OH) (5; L = HC[(CMe)(2,6-iPr2C6H3N)]2) has been prepared by the hydrolysis of LAlMeCl (4). For the preparation of 5, the reagents of KOH, water, and KH, as well as the two-phase ammonia/toluene system, were used. Further reactions of 5 with Cp2ZrMe2 (8) and Cp2ZrHCl in toluene lead to the intermolecular elimination of CH4 and H2 and the formation of mu-O-bridged dinuclear aluminum and zirconium complexes [LAlMe(mu-O)ZrMeCp2] (6) and [LAlMe(mu O)ZrClCp2] (7), respectively, in high yields. The crystal structure reveals that 5 is a monomer with terminal OH and Me groups. The X-ray structure analysis shows that 6 and 7 contain a bent Al-(mu-O)-Zr core with terminal Al-Me and Zr-Me or Zr Cl arrangements. The methylalumoxane (MAO)-activated compounds 6 and 7 exhibit high catalytic activity for the polymerization of ethylene. Under comparable polymerization conditions, the MAO/6 and MAO/7 catalyst systems show considerably higher activity and much lower MAO:catalyst ratios than that of MAO/8. PMID- 15755165 TI - Charge separation versus recombination in dye-sensitized nanocrystalline solar cells: the minimization of kinetic redundancy. AB - In this paper we focus upon the electron injection dynamics in complete dye sensitized nanocrystalline metal oxide solar cells (DSSCs). Electron injection dynamics are studied by transient absorption and emission studies of DSSCs and correlated with device photovoltaic performance and charge recombination dynamics. We find that the electron injection dynamics are dependent upon the composition of the redox electrolyte employed in the device. In a device with an electrolyte composition yielding optimum photovoltaic device efficiency, electron injection kinetics exhibit a half time of 150 ps. This half time is 20 times slower than that for control dye-sensitized films covered in inert organic liquids. This retardation is shown to result from the influence of the electrolyte upon the conduction band energetics of the TiO2 electrode. We conclude that optimum DSSC device performance is obtained when the charge separation kinetics are just fast enough to compete successfully with the dye excited-state decay. These conditions allow a high injection yield while minimizing interfacial charge recombination losses, thereby minimizing "kinetic redundancy" in the device. We show furthermore that the nonexponential nature of the injection dynamics can be simulated by a simple inhomogeneous disorder model and discuss the relevance of our findings to the optimization of both dye sensitized and polymer based photovoltaic devices. PMID- 15755167 TI - Synthesis of macrocycles via cobalt-mediated [2 + 2 + 2] cycloadditions. AB - We investigated the formation of macrocycles from alpha,omega-diynes in cobalt mediated co-cyclotrimerization reactions. Long-chain alpha,omega-diynes underwent metal-mediated [2 + 2 + 2] cycloadditions with nitriles, cyanamides, or isocyanates in the presence of CpCo(CO)2 (Cp = cyclopentadienide) to yield pyridine-containing macrocycles, i.e., meta- and para-pyridinophanes, such as 5m/5p, 35m/35p, and 41m/41p. The regioselectivity of these reactions was affected by the length and type of linker unit between the alkyne groups, as well as by certain stereoelectronic factors. An analogous alpha,omega-cyano-alkyne, 28, combined with an alkyne to yield two isomeric meta-pyridinophanes, such as 5m and 29m, and an ortho cycloadduct (benzannulation product), such as 29o. We developed a reaction protocol for these cobalt-based [2 + 2 + 2] cycloadditions that involves markedly improved conditions such that this process offers a convenient, flexible synthetic approach to macrocyclic pyridine-containing compounds. For example, diyne 6 reacted with p-tolunitrile in 1,4-dioxane to give 7p and 7m (7:1 ratio) in 87% yield at a moderate temperature of ca. 100 degrees C in 24 h without photoirradiation or syringe-pump addition. Isocyanates were also effective reactants, as exemplified by the formation of 44p almost exclusively (44p:44m > 50:1) in 64% yield from diyne 8 and 2-phenylethylisocyanate. By using this improved protocol we were able to co-cyclotrimerize long-chain alpha,omega diynes with alkynes in certain cases to demonstrate a successful macrocyclic variant of the Vollhardt reaction. For instance, diyne 6 reacted with dipropylacetylene to give paracyclophane 57p and benzannulene 57o (2:1 ratio) in 29% yield. PMID- 15755166 TI - Aqueous dispersions of single-wall and multiwall carbon nanotubes with designed amphiphilic polycations. AB - Poor solubility of single-walled and multiwalled carbon nanotubes (NTs) in water and organic solvents presents a considerable challenge for their purification and applications. Macromolecules can be convenient solubilizing agents for NTs and a structural element of composite materials for them. Several block copolymers with different chemical functionalities of the side groups were tested for the preparation of aqueous NT dispersions. Poly(N-cetyl-4-vinylpyridinium bromide-co N-ethyl-4-vinylpyridinium bromide-co-4-vinylpyridine) was found to form exceptionally stable NT dispersions. It is suggested that the efficiency of macromolecular dispersion agents for NT solubilization correlates with the topological and electronic similarity of polymer-NT and NT-NT interactions in the nanotube bundles. Raman spectroscopy and atomic force and transmission electron microcopies data indicate that the polycations are wrapped around NTs forming a uniform coating 1.0-1.5 nm thick. The ability to wind around the NT originates in the hydrophobic attraction of the polymer backbone to the graphene surface and topological matching. Tetraalkylammonium functional groups in the side chains of the macromolecule create a cloud of positive charge around NTs, which makes them hydrophilic. The prepared dispersions could facilitate the processing of the nanotubes into composites with high nanotube loading for electronic materials and sensing. Positive charge on their surface is particularly important for biological and biomedical applications because it strengthens interactions with negatively charged cell membranes. A high degree of spontaneous bundle separation afforded by the polymer coating can also be beneficial for NT sorting. PMID- 15755168 TI - Two-dimensional pressure-driven nanorod-to-nanowire reactions in Langmuir monolayers at room temperature. AB - Typical two-dimensional surface pressures prevalent within floating monolayer (Langmuir) films are on the order of a few dozen megapascals. One might expect, therefore, that some chemical reactions should be directly and strongly affected by this surface pressure, along with the well-known effect of pressure on the orientational order and the mere proximity of the molecules within the film. Here, we show that the two-dimensional surface pressure in Langmuir films provides a direct driving force, decreasing the activation energy of a cooperative oriented coalescence of ZnS nanorods into nanowires near room temperature. At low film surface pressure or low temperature, the nanorods do not react, while in solution, they react only above 140 degrees C, even though in all of these cases, the rods are organized in similar super-crystalline clusters. Electron microscopy and measurements of the reaction rates give a detailed picture of the order of the rods, their rearrangement, and their coalescence. PMID- 15755169 TI - Crystal structure and diffusion path in the fast lithium-ion conductor La(0.62)Li(0.16)TiO3. AB - We report the results of a neutron powder diffraction study of the La(0.62)Li(0.16)TiO3 perovskite that determined the diffusion path of lithium cations at room temperature. At 77 K, the Li cations are located at the 2c site (Wycoff notation of the Cmmm space group) on the (002) La deficient layer, while, at room temperature, they are spread over a wide area and migrate following the 2c-4f-2c or 2c-2d-2c tie line on the (002) layer. The probability density of Li cations has a minimum between the 2c and 4f or between the 2c and 2d positions on the diffusion path in contrast to the previous reports where the bottleneck has been thought to be located at the 2c, 2d, and 4f positions. On the basis of the present structural model, the Li-cation conductivity is discussed in terms of a two-dimensional bond-percolation model for Li-cation diffusion. It was found that the vacancy at the La site is essential for the Li-cation conduction. PMID- 15755170 TI - Stable transition metal complexes of an all-metal antiaromatic molecule (Al4Li4): role of complexations. AB - We propose for the first time a few examples of stable transition metal complexes of an all-metal antiaromatic molecule Al4Li4. We demonstrate that these all-metal species can be stabilized by complexation with 3d transition metals very similar to their organic counterpart, C4H4. Complexation to transition metal ions reduces the frontier orbital energies and introduces aromatic charactersitics. We consider a series of such complexes: [eta4-(Al4Li4)-Fe(CO)3, eta2,sigma2-(Al4Li4) Ni, and (Al4Li4)2Ni] and compare and contrast their energetics with their organometallic counterparts. Fragmentation energy, orbital correlation energy analysis, and the nucleus-independent chemical shift (NICS) values support the complexation-induced stabilities in these systems. PMID- 15755171 TI - Lanthanide clusters with internal Ln ions: highly emissive molecules with solid state cores. AB - "Er(SePh)(2.5)I(0.5)" reacts with elemental S to give (THF)10Er6S6I6, a double cubane cluster with one face of the Er4S4 cube capped by an additional Er2S2. Reactions with a mixture of elemental S/Se results in the formation of (THF)14Er10S6(Se2)6I6, a cluster composed of an Er6S6 double cubane core, with two "Er2(Se2)3" units condensed onto opposing rectangular sides of the Er6S6 fragment. This deposition of Er2Se6 totally encapsulates the two central Er with chalcogen atoms (4 S, 4 Se) and excludes neutral THF donors or iodides from the two primary coordination spheres. The Er10 compound is the first lanthanide cluster to contain internal, chalcogen encapsulated Ln. This cluster shows strong fluorescence at 1544 nm with a measured decay time of 3 ms and an estimated quantum efficiency of 78%, which is comparable to Er doped solid-state materials. The unusual fluorescence spectral properties of (THF)14Er10S6Se12I6 are unprecedented for a molecular Er complex and are attributed to the low phonon energy host environment provided by the I-, S(2-), and Se2(2-) ligands. PMID- 15755172 TI - Metallurgy in a beaker: nanoparticle toolkit for the rapid low-temperature solution synthesis of functional multimetallic solid-state materials. AB - Intermetallic compounds and alloys are traditionally synthesized by heating mixtures of metal powders to high temperatures for long periods of time. A low temperature solution-based alternative has been developed, and this strategy exploits the enhanced reactivity of nanoparticles and the nanometer diffusion distances afforded by binary nanocomposite precursors. Prereduced metal nanoparticles are combined in known ratios, and they form nanomodulated composites that rapidly transform into intermetallics and alloys upon heating at low temperatures. The approach is general in terms of accessible compositions, structures, and morphologies. Multiple compounds in the same binary system can be readily accessed; e.g., AuCu, AuCu3, Au3Cu, and the AuCu-II superlattice are all accessible in the Au-Cu system. This concept can be extended to other binary systems, including the intermetallics FePt3, CoPt, CuPt, and Cu3Pt and the alloys Ag-Pt, Au-Pd, and Ni-Pt. The ternary intermetallic Ag2Pd3S can also be rapidly synthesized at low temperatures from a nanocomposite precursor comprised of Ag2S and Pd nanoparticles. Using this low-temperature solution-based approach, a variety of morphologically diverse nanomaterials are accessible: surface-confined thin films (planar and nonplanar supports), free-standing monoliths, nanomesh materials, inverse opals, and dense gram-scale nanocrystalline powders of intermetallic AuCu. Importantly, the multimetallic materials synthesized using this approach are functional, yielding a room-temperature Fe-Pt ferromagnet, a superconducting sample of Ag2Pd3S (Tc = 1.10 K), and a AuPd4 alloy that selectively catalyzes the formation of H2O2 from H2 and O2. Such flexibility in the synthesis and processing of functional intermetallic and alloy materials is unprecedented. PMID- 15755173 TI - Interaction of a bulky N-heterocyclic carbene ligand with Rh(I) and Ir(I). Double C-H activation and isolation of bare 14-electron Rh(III) and Ir(III) complexes. AB - Reactivity and structural studies of unusual rhodium and iridium systems bearing two N-heterocyclic carbene (NHC) ligands are presented. These systems are capable of intramolecular C-H bond activation and lead to coordinatively unsaturated 16 electron complexes. The resulting complexes can be further unsaturated by simple halide abstraction, leading to 14-electron species bearing an all-carbon environment. Saturation of the vacant sites in the 16- and 14-electron complexes with carbon monoxide permits a structural comparison. DFT calculations show that these electrophilic metal centers are stabilized by pi-donation of the NHC ligands. PMID- 15755174 TI - A tryptophan-containing open-chain framework for tuning a high selectivity for Ca2+ and 13C NMR observation of a Ca2+-indole interaction in aqueous solution. AB - Two molecular architectures featuring the cation-responsive tryptophan indole were designed and investigated for the development of a novel fluorescent chemosensor for Ca2+. We observed that the Trp-based open-framework chemosensor EW2 exhibits remarkable selectivity for Ca2+ over Mg2+, Ba2+, K+, Na+, and Li+ in water between pH 4.6 and 7.0 on the basis of Ca2+-induced high fluorescence enhancement of the Trp residue. A combined 13C NMR and CD spectroscopic study has demonstrated a dynamic reorientation of the indole ring due to the cation-indole interaction accompanying the Ca2+-induced dramatic fluorescence enhancement. The results suggest that the highly sensitive, metal-ion-dependent Trp indolyl C(3) chemical shifts may serve as a promising indicator for monitoring metal ion indole noncovalent interaction in solution. PMID- 15755175 TI - Spectroscopic and density functional studies of the red copper site in nitrosocyanin: role of the protein in determining active site geometric and electronic structure. AB - The electronic structure of the red copper site in nitrosocyanin is defined relative to that of the well understood blue copper site of plastocyanin by using low-temperature absorption, circular dichroism, magnetic circular dichroism, resonance Raman, EPR and X-ray absorption spectroscopies, combined with DFT calculations. These studies indicate that the principal electronic structure change in the red copper site is the sigma rather than the pi donor interaction of the cysteine sulfur with the Cu 3d(x2-y2) redox active molecular orbital (RAMO). Further, MCD data show that there is an increase in ligand field strength due to an increase in coordination number, whereas resonance Raman spectra indicate a weaker Cu-S bond. The latter is supported by the S K-edge data, which demonstrate a less covalent thiolate interaction with the RAMO of nitrosocyanin at 20% relative to plastocyanin at 38%. EXAFS results give a longer Cu-S(Cys) bond distance in nitrosocyanin (2.28 A) compared to plastocyanin (2.08 A) and also show a large change in structure with reduction of the red copper site. The red copper site is the only presently known blue copper-related site with an exogenous water coordinated to the copper. Density functional calculations reproduce the experimental properties and are used to determine the specific protein structure contributions to exogenous ligand binding in red copper. The relative orientation of the CuNNS and the CuSC(beta) planes (determined by the protein sequence) is found to be key in generating an exchangeable coordination position at the red copper active site. The exogenous water ligation at the red copper active site greatly increases the reorganization energy (by approximately 1.0 eV) relative to that of the blue copper protein site, making the red site unfavorable for fast outer-sphere electron transfer, while providing an exchangeable coordination position for inner-sphere electron transfer. PMID- 15755176 TI - O-atom transport catalysis by atomic cations in the gas phase: reduction of N2O by CO. AB - Atomic cations (26), M+, have been shown to lie within a thermodynamic window for O-atom transport catalysis of the reduction of N2O by CO and have been checked for catalytic activity at room temperature with kinetic measurements using an inductively-coupled plasma/selected-ion flow tube (ICP/SIFT) tandem mass spectrometer. Only 10 of these 26 atomic cations were seen to be catalytic: Ca+, Fe+, Ge+, Sr+, Ba+, Os+, Ir+, Pt+, Eu+, and Yb+. The remaining 16 cations that lie in the thermodynamic window (Cr+, Mn+, Co+, Ni+, Cu+, Se+, Mo+, Ru+, Rh+, Sn+, Te+, Re+, Pb+, Bi+, Tm+, and Lu+) react too slowly at room temperature either in the formation of MO+ or in its reduction by CO. Many of these reactions are known to be spin forbidden and a few actually may lie outside the thermodynamic window. A new measure of efficiency is introduced for catalytic cycles that allows the discrimination between catalytic cations on the basis of the efficiencies of the two legs of the catalytic cycle. Also, a potential-energy landscape is computed for the reduction of N2O by CO catalyzed by Fe+(6D) that vividly illustrates the operation of an ionic catalyst. PMID- 15755177 TI - On the mechanism of hydrophobic association of nanoscopic solutes. AB - The hydration behavior of two planar nanoscopic hydrophobic solutes in liquid water at normal temperature and pressure is investigated by calculating the potential of mean force between them at constant pressure as a function of the solute-solvent interaction potential. The importance of the effect of weak attractive interactions between the solute atoms and the solvent on the hydration behavior is clearly demonstrated. We focus on the underlying mechanism behind the contrasting results obtained in various recent experimental and computational studies on water near hydrophobic solutes. The length scale where crossover from a solvent separated state to the contact pair state occurs is shown to depend on the solute sizes as well as on details of the solute-solvent interaction. We find the mechanism for attractive mean forces between the plates is very different depending on the nature of the solute-solvent interaction which has implications for the mechanism of the hydrophobic effect for biomolecules. PMID- 15755178 TI - Asymmetric induction during Yang cyclization of alpha-oxoamides: the power of a covalently linked chiral auxiliary is enhanced in the crystalline state. AB - Gamma-hydrogen abstraction has been revealed to be the primary photoprocess in the crystalline state of alpha-oxoamides through photochemical and X-ray structural studies. The outstanding ability of a covalent chiral auxiliary in generating asymmetric induction in the photoproduct beta-lactam has been established with 10 examples. We have shown that the crystal lattice preorganizes the reactant molecules toward a single diastereomer of the beta-lactam and prevents large motions of the 1,4-diradical intermediate that would result in the loss of stereochemical memory. A rare single-crystal-to-single-crystal transformation path of one of the examples investigated establishes the direct correlation between the stereochemistries of the reactant and the product. PMID- 15755179 TI - Macrophomate synthase: QM/MM simulations address the Diels-Alder versus Michael Aldol reaction mechanism. AB - Macrophomate synthase (MPS) of the phytopathogenic fungus Macrophoma commelinae catalyzes the transformation of 2-pyrone derivatives to the corresponding benzoate analogues. The transformation proceeds through three separate chemical reactions, including decarboxylation of oxalacetate to produce pyruvate enolate, two C-C bond formations between 2-pyrone and pyruvate enolate that form a bicyclic intermediate, and final decarboxylation with concomitant dehydration. Although some evidence suggests that the second step of the reaction catalyzed by MPS is a Diels-Alder reaction, definite proof that the C-C bond formations occur via a concerted mechanism is still required. An alternative route for formation of the C-C bonds is a stepwise Michael-aldol reaction. In this work, mixed quantum and molecular mechanics (QM/MM) combined with Monte Carlo simulations and free-energy perturbation (FEP) calculations were performed to investigate the relative stabilities of the transition states (TS) for both reaction mechanisms. The key results are that the Diels-Alder TS model is 17.7 and 12.1 kcal/mol less stable than the Michael and aldol TSs in the stepwise route, respectively. Therefore, this work indicates that the Michael-aldol mechanism is the route used by MPS to catalyze the second step of the overall transformation, and that the enzyme is not a natural Diels-Alderase, as claimed by Ose and co-workers (Nature 2003, 422, 185-189; Acta Crystallogr. 2004, D60, 1187-1197). A modified link-atom treatment for the bonds at the QM/MM interface is also presented. PMID- 15755180 TI - Limited flexibility of lactose detected from residual dipolar couplings using molecular dynamics simulations and steric alignment methods. AB - The conformational flexibility of lactose in solution has been investigated by residual dipolar couplings (RDCs). One-bond carbon-proton and proton-proton coupling constants have been measured in two oriented media and interpreted in combination with molecular dynamics simulations (MD). Two different approaches, known as PALES (Zweckstetter et al., J. Am. Chem. Soc. 2000, 122, 3791-3792) and TRAMITE (Azurmendi et al., J. Am. Chem. Soc. 2002, 124, 2426-2427), have been used to determine the alignment tensor from a shape-induced alignment model with the oriented medium. The steric alignment of the structures from several MD trajectories has provided ensemble averaged RDCs that have been compared with the experimental ones. The obtained results reveal the almost exclusive presence of a major low energy region defined as syn-phi/syn-psi (> 97%), for which sampling occurs in a dynamic manner. This result satisfactorily agrees with that determined by standard NOE-based methods. PMID- 15755181 TI - Electrochemical deposition of mesoporous nickel hydroxide films from dilute surfactant solutions. AB - A series of micelle-templated mesoporous nickel hydroxide films were prepared by electrochemical deposition from dilute surfactant solutions by using different types of template and by varying plating solvent composition. Lamellar mesostructured Ni(OH)2 films are obtained with only anionic surfactant sodium dodecyl sulfate (SDS) as the template. In particular, a unique cooperative assembly fashion, that is, the combination between Ni2+ and a complex composed of the primary template SDS and a cosurfactant, such as triblock poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) copolymers and poly(ethylene glycol), was explored, by which two-dimensional hexagonal mesoporous Ni(OH)2 films were electrodeposited. Meanwhile, the deposition medium also plays a crucial role in determining the mesostructure of Ni(OH)2 films. For the composite nickel hydroxide films deposited from aqueous solution or dilute aqueous solution of ethylene glycol (<20 wt %) in the presence of SDS or the SDS poly(alkylene oxide) polymer complexes, a mixed lamellar phase with d(001) = 37.4 A and d(001) = 28.5 A was obtained. However, single lamellar phase with d(001) = 37.4 A was electrodeposited from concentrated aqueous solutions of ethylene glycol (> or = 20 wt %). Furthermore, such deposition baths have access to hexagonal mesoporous nickel hydroxide films with d(100) = 37.4 A at 70 degrees C with the SDS-poly(alkylene oxide) polymer complexes as the templates. Within the potential window for Ni(OH)2, the morphology and quality of mesostructured films are significantly dependent on the deposition potential, while the mesostructures of the composite films always remain unchanged. PMID- 15755182 TI - 2,6-Diarylnaphtho[1,8-bc:5,4-b'c']dithiophenes as new high-performance semiconductors for organic field-effect transistors. AB - A series of 2,6-diaryl-substituted naphtho[1,8-bc:5,4-b'c']dithiophene derivatives 2-6, whose aryl groups include 5-hexyl-2-thienyl, 2,2'-bithiophen-5 yl, phenyl, 2-naphthyl, and 4-biphenylyl, was synthesized by the palladium catalyzed Suzuki-Miyaura coupling and utilized as active layers of organic field effect transistors (OFETs). All devices fabricated using vapor-deposited thin films of these compounds showed typical p-type FET characteristics. The mobilities are relatively good and widely range from 10(-4) to 10(-1) cm2 V(-1) s(-1), depending on the substituent groups. Among them, the mobilities of the devices using films of 3-5 tend to increase with the increasing temperature of the Si/SiO2 substrate during film deposition. In particular, the device based on the naphthyl derivative 5, when fabricated at 140 degrees C, marked a high mobility of 0.11 cm2 V(-1) s(-1) with an on/off ratio of 10(5), which is a top class of performance among organic thin-film transistors. Studies of X-ray diffractograms (XRDs) have revealed that the film of 4 and 5 is composed of two kinds of crystal grains with different phases, so-called "single-crystal phase" and "thin film phase", and that the proportion of the thin film phase increases with an increase of the substrate temperature. In the thin film phase the assembled molecules stand nearly upright on the substrate in such a way as to be favorable to carrier migration. PMID- 15755184 TI - Computational rationalization of the dependence of the enantioselectivity on the nature of the catalyst in the vanadium-catalyzed oxidation of sulfides by hydrogen peroxide. AB - A computational study with the IMOMM(Becke3LYP:MM3) method is carried out on the mechanism of the enantioselective reaction of complex V(O)(L)(OOH), L= bulky tridentate Schiff base, and bis(tert-butyl) disulfide. The reaction with a given L ligand A is first systematically studied: different conformers of the catalyst are optimized, and the large number of associated transition states are systematically searched. The study is then extended to the geometry optimization of selected transition states associated to other ligands B, C, and D, similar to A but differing in the nature of certain substituents R1, R2, R3. The experimental trends in selectivity for catalysts based on ligands A to D are faithfully reproduced by the calculations. Analysis of the computational results leads finally to the formulation of a simple model that can explain one of the most remarkable aspect of this reaction, namely the large effect on enantioselectivity of ligands seemingly far from each other in the catalyst. PMID- 15755183 TI - Local and global effects of metal binding within the small subunit of ribonucleotide reductase. AB - Each beta-protomer of the small betabeta subunit of Escherichia coli ribonucleotide reductase (R2) contains a binuclear iron cluster with inequivalent binding sites: Fe(A) and Fe(B). In anaerobic Fe(II) titrations of apoprotein under standard buffer conditions, we show that the majority of the protein binds only one Fe(II) atom per betabeta subunit. Additional iron occupation can be achieved upon exposure to O2 or in high glycerol buffers. The differential binding affinity of the A- and B-sites allows us to produce heterobinuclear Mn(II)Fe(II) and novel Mn(III)Fe(III) clusters within a single beta-protomer of R2. The oxidized species are produced with H2O2 addition. We demonstrate that no significant exchange of metal occurs between the A- and B-sites, and thus the binding of the first metal is under kinetic control, as has been suggested previously. The binding of first Fe(II) atom to the active site in a beta protomer (betaI) induces a global protein conformational change that inhibits access of metal to the active site in the other beta-protomer (betaII). The binding of the same Fe(II) atom also induces a local effect at the active site in betaI-protomer, which lowers the affinity for metal in the A-site. The mixed metal FeMn species are quantitatively characterized with electron paramagnetic resonance spectroscopy. The previously reported catalase activity of Mn2(II)R2 is shown not to be associated with Mn. PMID- 15755185 TI - Enhanced fluorescence cyanide detection at physiologically lethal levels: reduced ICT-based signal transduction. AB - Three water-soluble fluorescent probes have been specifically designed to determine free cyanide concentrations up to physiologically lethal levels, >20 microM. The probes have been designed in such a way as to afford many notable sensing features, which render them unique with regard to signal transduction, photophysical characteristics, and their application to physiological cyanide determination and safeguard. The probes are readily able to reversibly bind free aqueous cyanide with dissociation constants around 4 microM3. Subsequent cyanide binding modulates the intramolecular charge transfer within the probes, a change in the electronic properties within the probes, resulting in enhanced fluorescence optical signals as a function of increased solution cyanide concentration. The ground-state chelation with cyanide produces wavelength shifts, which also enable the probes to sense cyanide in both an excitation and emission ratiometric manner, in addition to enhanced fluorescence signaling. This has enabled a generic cyanide sensing platform to be realized that is not dependent on fluorescent probe concentration, probe photodegradation, or fluctuations in the intensity of any employed excitation sources, ideal for remote cyanide sensing applications. Further, the >600 nm fluorescence emission of the probes potentially allows for enhanced fluorescence ratiometric cyanide sensing in the optical window of tissues and blood, facilitating their use for the transdermal monitoring of cyanide for mammalian safeguard or postmortem in fire victims, both areas of active research. PMID- 15755188 TI - Are cognitively impaired older adults able to vote? PMID- 15755189 TI - Reddish-brown itchy papule on leg. No substance is draining from the lesion. PMID- 15755190 TI - Psychoses in late life: evaluation and management of disorders seen in primary care. AB - As the ranks of Baby Boomers move into later life and develop age-related cognitive conditions, an increased incidence in the number of psychotic conditions is expected. When psychotic conditions occur after age 60, the causes and management pose challenges for clinicians caring for the older adult. This article will review the differential diagnosis and management of psychoses in late life. PMID- 15755191 TI - Preventive services: immunization and chemoprevention. PMID- 15755192 TI - Investigation of tetherable distilbazolium compounds as fluorescent probes in nanostructured silica sol-gel materials. AB - Distibazolium dyes are investigated by steady-state and time-resolved fluorescence techniques in a series of low- and high-viscosity polar solvents and in a silica sol-gel matrix. In all solvents and the sol-gel matrix, an interplay of photoinduced switching between different cis-trans isomers and solvation dynamics is observed. Even in a viscous solution (glycerol) and in silica gel, cis-trans isomerization is solvent-controlled. Whereas in glycerol the solvation results in a time-dependent fluorescence Stokes shift, the solvation-induced spectral heterogeneity in silica gel is mostly static, possibly due to a close proximity of dye molecules to the silica surfaces of the nanopores. Compared to low-viscosity solvents, where the cis-trans isomerization process takes place with a solvent-dependent rate on the timescale of about 120-150 ps, it slows down to about 1100-1400 ps in glycerol and about 1500 ps in a sol-gel matrix. Additionally, fluorescence kinetics of the dyes in the sol-gel reveals the presence of a range of different "frozen-in" conformers exhibiting a broad spectrum of lifetimes from 20 to 300 ps. PMID- 15755193 TI - Applications of free-electron lasers in the biological and material sciences. AB - Free-Electron Lasers (FELs) collectively operate from the terahertz through the ultraviolet range and via intracavity Compton backscattering into the X-ray and gamma-ray regimes. FELs are continuously tunable and can provide optical powers, pulse structures and polarizations that are not matched by conventional lasers. Representative research in the biological and biomedical sciences and condensed matter and material research are described to illustrate the breadth and impact of FEL applications. These include terahertz dynamics in materials far from equilibrium, infrared nonlinear vibrational spectroscopy to investigate dynamical processes in condensed-phase systems, infrared resonant-enhanced multiphoton ionization for gas-phase spectroscopy and spectrometry, infrared matrix-assisted laser-desorption-ionization and infrared matrix-assisted pulsed laser evaporation for analysis and processing of organic materials, human neurosurgery and ophthalmic surgery using a medical infrared FEL and ultraviolet photoemission electron microscopy for nanoscale characterization of materials and nanoscale phenomena. The ongoing development of ultraviolet and X-ray FELs are discussed in terms of future opportunities for applications research. PMID- 15755194 TI - Exposure geometry and spectral environment determine photobiological effects on the human eye. AB - Photobiological effects upon the human retina, cornea and lens are highly dependent on the optical exposure geometry as well as spectral characteristics of the exposure. The organ of sight is exquisitely sensitive to light because it performs well in very low nighttime illumination levels and yet it also must adapt to extremely bright environments where light exposures are greater by many orders of magnitude. The eye has evolved to protect itself reasonably well against excessive exposure in bright environments. The retina is minimally exposed in extremely bright environments and the cornea and lens are surprisingly well protected in harsh environments. Although these protective mechanisms are good, they are not perfect and adverse changes from both acute and chronic exposures to sunlight still exist. The geometrical protective factors must be understood and appreciated whenever assessing potential adverse effects of environmental UV radiation and light on ocular structures. These natural ocular protective factors also work with the ever-changing spectrum of sunlight and the different spectral distribution of light and UV radiation across the eye's field of view. Spectral characteristics of the ocular media are also important. One can visualize a series of intraocular color filters that progressively filter shorter wavelengths and thereby aid in color vision, reduce the impact of chromatic aberrations and significantly reduce the optical radiation hazards to the lens and retina. PMID- 15755197 TI - Analysis of 6000 skin biopsies of the national leprosy control program in Mexico. AB - Six thousand skin biopsy specimens taken from April 1978 to January 2002 under conditions as specified by the National Leprosy Control Program (NLCP), were analyzed to obtain information about the work of the program and contribute to the knowledge of this illness in the Mexico. Six-thousand request forms for histologic exam of the NLCP were reviewed. Sixty-two percent of the requests had all the required information and in 38% one or more data items were omitted. The age range was 2 to 98 yrs with a median of 50 yrs; a small number of cases was observed in the age group of 0 to 14 yrs, and the peak was in the age group of 41 to 50 yrs. Of the 6000 biopsies, 3693 were classified. Polar lepromatous (LL) was the most common form of the disease, in 60.3% of cases. Twice as many cases were multibacillary leprosy (MB) as paucibacillary (PB). MB predominated in males, and PB predominated in females. The Cohen's kappa index (kappa) of clinical histological agreement was 0.202 (95% CI 0.184-0.219) and showed a poor grade of agreement between clinical and histologic diagnosis, with a level of significance of 0.05 (p <0.001). The results may indicate the end of leprosy in Mexico, a country in which the national goal of elimination was reached in 1994, with a prevalence since the year 2000 of 0.17/10 000. PMID- 15755198 TI - The dynamics of stigma in leprosy. AB - Leprosy in Nepal is a stigmatizing disease. This paper explores the different coping strategies employed by people affected by leprosy to manage stigma. It is based on a qualitative study conducted in the eastern part of Nepal. It will show that a difference exists between experienced stigma and the anticipation of stigma. Both types of stigma result in different coping strategies. In managing stigma people go through different phases. This paper will show that stigma is a dynamic process, and I will elaborate on the concealment cycle, as developed by Hyland, to produce a more detailed understanding of the stigmatization process in Nepal. Doing so, it highlights the importance of a mutual concealment phase and the importance of triggers to exposure and discrimination. Changing from one phase to a subsequent phase in the stigmatization process is always triggered. It highlights further, that even within the same culture and even the same village, social differentiation makes a significant difference on the impact of stigma and the coping strategies employed in managing stigma. Stigma enforces already existing inequalities in social class, gender, and age. PMID- 15755195 TI - The global threat of counterfeit drugs: why industry and governments must communicate the dangers. PMID- 15755199 TI - A comparison of economic aspects of hospitalization versus ambulatory care in the management of neuritis occurring in lepra reaction. AB - Neuritis is one of the important causes of deformities and disabilities in leprosy. Neuritis has been managed both in the field and in hospital. This study was done to compare the economic aspects of cost of ambulatory vs in-patient management of neuritis in leprosy. The quality of life of the affected patients and the clinical improvement in the 2 groups were also studied. Twenty six patients fulfilling the study criteria were randomized into the ambulatory and in patient group (13 in each group). The primary outcome examined was cost, in various categories; the secondary outcomes included pre- and post-treatment comparison of Quality of Life (QOL) scores and tests of sensory and motor function. The direct and indirect medical costs incurred by patients in the hospitalized group were higher than those patients in the ambulatory group. The difference in the direct medical costs between the two groups was Rs. 9110.5, and the extra direct non medical costs incurred by patients in the hospitalized group was Rs. 888.50 because of more frequent visits of family members. A greater percentage of ambulatory than in-patients returned to work in 28; (28/7) -> 4; and (28/4) --> 7. In this article, the author reviews the evidence for this model, considers alternative accounts, and proposes a simple and empirically motivated revision to the model that (a) accommodates conflicting results, (b) provides a novel account of the ties effect, and (c) makes new and nonintuitive predictions for the factoring operation (e.g., factoring of 28 into 4 and 7). The author reports 3 experiments designed to test these predictions and discusses implications for arithmetic instruction. PMID- 15755244 TI - Integrating novel dimensions to eliminate category exceptions: when more is less. AB - Category learning can be characterized as a process of discovering the dimensions that represent stimuli efficiently and effectively. Categories that are overlapping when represented in 1 dimensionality may be separate in a higher dimensional cue set. The authors report 2 experiments in which participants were shown an additional cue after learning to use 2 imperfect cues. The results revealed that participants can integrate new information into their categorization cue set. The authors discovered wide individual differences, however, with many participants favoring simpler, but less accurate, cue sets. Some participants demonstrated the ability to discard information previously used when new, more accurate information was introduced. The categorization model RASHNL (J. K. Kruschke & M. K. Johansen, 1999) gave qualitatively accurate fits of the data. PMID- 15755245 TI - The origin of exemplar effects in rule-driven categorization. AB - S. W. Allen and L. R. Brooks (1991) have shown that exemplar memory can affect categorization even when participants are provided with a classification rule. G. Regehr and L. R. Brooks (1993) argued that stimuli must be individuated for such effects to occur. In this study, the authors further analyze the conditions that yield exemplar effects in this rule application paradigm. The results of Experiments 1-3 show that interchangeable attributes, which are not part of the rule, influence categorization only when attention is explicitly drawn on them. Experiment 4 shows that exemplar effects can occur in an incidental learning condition, whether stimulus individuation is preserved or not. The authors conclude that the influence of exemplar learning in rule-driven categorization stems from the attributes specified in the rule or in the instructions, not from the stimulus gestalts. PMID- 15755246 TI - An exemplar account of the bow and set-size effects in absolute identification. AB - The extended generalized context model for response times (K. Lamberts, 2000) was designed to account for choice proportions and response times in perceptual categorization. In this article, the hypothesis that the model also offers an account of accuracy and response times in absolute identification was investigated. The model was applied to the data from 2 absolute identification experiments and provided a good account of the bow and the set-size effects in accuracy and response time data from individual participants, including the response time distributions for individual stimuli. The model applications demonstrated that exemplar-based process models offer a viable account of absolute identification data. PMID- 15755247 TI - Examining the time course of indexical specificity effects in spoken word recognition. AB - Variability in talker identity and speaking rate, commonly referred to as indexical variation, has demonstrable effects on the speed and accuracy of spoken word recognition. The present study examines the time course of indexical specificity effects to evaluate the hypothesis that such effects occur relatively late in the perceptual processing of spoken words. In 3 long-term repetition priming experiments, the authors examined reaction times to targets that were primed by stimuli that matched or mismatched on the indexical variable of interest (either talker identity or speaking rate). Each experiment was designed to manipulate the speed with which participants processed the stimuli. The results demonstrate that indexical variability affects participants' perception of spoken words only when processing is relatively slow and effortful. PMID- 15755249 TI - Sticky rules: integration between abstract rules and specific actions. AB - The authors manipulated repetitions and/or changes of abstract response rules and the specific stimulus- response (S-R) associations used under these rules. Experiments 1 and 2, assessing trial-to-trial priming effects, showed that repetition of complete S-R couplings produced only benefits when the rule also repeated (i.e., rule-S-R conjunctions) but costs when identical S-R couplings repeated while rules changed. In Experiments 3 and 4, the authors manipulated amount of experience with specific rule-S-R conjunctions and demonstrated integration between rules and S-R couplings in terms of cumulative practice effects. However, unlike short-term priming effects, cumulative practice supported generalization of experience with specific S-R couplings across rule boundaries (Experiment 4). Results are discussed in terms of constraints on models of hierarchical control and in terms of qualitatively different ways in which people profit from very recent experiences (i.e., all-or-none access to working memory representations) versus cumulative experience (i.e., similarity based retrieval from long-term memory). PMID- 15755248 TI - The "one-shot" hypothesis for context storage. AB - In 3 experiments motivated by the implicit memory literature, the authors investigated the effects of different strengthening operations on the list strength effect (LSE) for explicit free recall, an effect posited by R. M. Shiffrin, R. Ratcliff, and S. E. Clark (1990) to be due to context cuing. According to the one-shot hypothesis, a fixed amount of context is stored when an item is studied for at least 1 or 2 s. Beyond the initial context storage, increases in study time or different orienting tasks do not influence the amount of context that is stored, and thus only spaced repetitions should produce a positive LSE. Consistent with prior findings, spaced repetitions always produced a positive LSE, but increases in depth of processing, study time, and massed repetitions did not. A model implements the one-shot hypothesis, and a role for context storage as a link between episodic and semantic memory is discussed. PMID- 15755251 TI - Regularization in short-term memory for serial order. AB - Previous research has shown that short-term memory for serial order can be influenced by background knowledge concerning regularities of sequential structure. Specifically, it has been shown that recall is superior for sequences that fit well with familiar sequencing constraints. The authors report a corresponding effect pertaining to serial recall errors. Undergraduate participants performed immediate serial recall on sequences of pseudowords generated on the basis of an artificial grammar. After extensive experience with this material, recall errors displayed a bias toward regularizing responses, response sequences more probable, with respect to the artificial grammar, than the originally presented stimulus sequence. This regularization effect squares well with recent trace redintegration and Bayesian models of serial recall, and appears to represent an analog of the schema-based error patterns observed in other domains of memory. PMID- 15755252 TI - Memory for narrative and expository text: independent influences of semantic associations and text organization. AB - The author examined memory for text in terms of the independent influences of semantic knowledge associations and text organization. Semantic associations were operationalized as the semantic relatedness between individual text concepts and the text as a whole and assessed with latent semantic analysis. The author assessed text organization by simulating comprehension with the construction integration model. Text organization consistently accounted for unique variance in recall. Semantic associations strongly predicted expository recall and predicted narrative recall significantly but to a lesser extent, even when the familiarity of the narrative content was manipulated. Results suggest that prior semantic associations and novel associations in the text structure influence memory independently, and that these influences can be affected by text genre. PMID- 15755253 TI - An alternative perspective on von Winterfeldt et al.'s (1997) test of consequence monotonicity. AB - D. von Winterfeldt, N.-K. Chung, R. D. Luce, and Y. Cho (1997) provided several tests for consequence monotonicity of choice or judgment, using certainty equivalents of gambles. The authors reaxiomatized consequence monotonicity in a probabilistic framework and reanalyzed von Winterfeldt et al.'s main experiment via a bootstrap method. Their application offers new insights into consequence monotonicity as well as into von Winterfeldt et al.'s 3 experimental paradigms: judged certainty equivalents (JCE), QUICKINDIFF, and parameter estimation by sequential testing (PEST). For QUICKINDIFF, the authors found no indication of violations of "random consequence monotonicity." This sharply contrasts the findings of von Winterfeldt et al., who concluded that axiom violations were the most pronounced under that procedure. The authors found potential evidence for violations in JCE and certainty equivalents derived from PEST. PMID- 15755254 TI - Predicting without modeling: a critique of Trabasso and Bartolone (2003). AB - T. Trabasso and J. Bartolone (2003) used a computational model of narrative text comprehension to account for empirical findings. The authors show that the same predictions are obtained without running the model. This is caused by the model's computational setup, which leaves most of the model's input unchanged. PMID- 15755256 TI - Structural aspects of histone H1-DNA complexes and their relation to transfection efficiency. AB - During transfection, polycation-DNA complexes are normally diluted by the transfection medium, which often contains salt in the physiological concentration range and serum. It is not exactly known to what extent this dilution step influences the properties of the complexes, which in turn influence the transfection efficiency. In order to gain more insight into the size-structure transfection activity relationship, we prepared histone H1-DNA complexes in NaCl solutions at various concentrations known to determine the size and structure of the resulting complexes. We characterized the complexes by physicochemical methods. Fluorescence correlation spectroscopy enabled relative measurements of complex sizes even under physiological conditions. The different appearances of the complexes were correlated with their transfection efficiency. When transfection was performed by dilution of the complexes in cell-cultivation media, the initial structure of H1-DNA complexes preformed under distinct salt conditions had no significant influence on the transfection efficiency. The dilution of the preformed complexes with cell-cultivation medium resulted in re formation and aggregation of the complexes. The addition of the complexes to the cells without cell-cultivation medium, however, showed a direct correlation between the size of the complexes and the transfection efficiency (correlation coefficient 0.91). Small complexes did not contribute to the transfection. PMID- 15755257 TI - Blood haemoglobin is an independent predictor of B-type natriuretic peptide (BNP). AB - BNP (B-type natriuretic peptide) and anaemia are both associated with adverse outcome in patients with chronic heart failure. Whether low haemoglobin levels are independently predictive of elevated BNP levels in subjects without heart failure is unknown. In the present study, we examined the relationship between haemoglobin and BNP levels in 234 patients with suspected coronary heart disease without a history of chronic heart failure, adjusting for known predictors of BNP levels. By univariate analysis, haemoglobin levels were inversely related to logarithmically transformed BNP values (r = -0.30, P < 0.0001). After adjustment for patient age, gender, body mass index, history of myocardial infarction, use of diuretics, angiotensin-converting enzyme inhibitors and beta-blockers, estimated creatinine clearance rate, extent of coronary disease, left ventricular ejection fraction and left ventricular end-diastolic pressure, blood haemoglobin remained an independent predictor of plasma BNP (standardized beta-coefficient = 0.253, P < 0.0001). A similar relationship was observed between haematocrit and BNP (standardized beta-coefficient -0.215, P < 0.0001). We conclude that haemoglobin levels are independently predictive of plasma BNP levels in patients with suspected coronary heart disease without heart failure. Anaemia may contribute to elevated BNP levels in the absence of heart failure, and may represent an important confounder of the relationship between BNP, cardiac function and prognosis. PMID- 15755258 TI - Membrane activity of the phospholipase C-delta1 pleckstrin homology (PH) domain. AB - PH-PLCdelta1 [the PH domain (pleckstrin homology domain) of PLCdelta1 (phospholipase C-delta1)] is among the best-characterized phosphoinositide binding domains. PH-PLCdelta1 binds with high specificity to the headgroup of PtdIns(4,5)P2, but little is known about its interfacial properties. In the present study, we show that PH-PLCdelta1 is also membrane-active and can insert significantly into PtdIns(4,5)P2-containing monolayers at physiological (bilayer equivalent) surface pressures. However, this membrane activity appears to involve interactions distinct from those that target PH-PLCdelta1 to the PtdIns(4,5)P2 headgroup. Whereas the majority of PtdIns(4,5)P2-bound PH-PLCdelta1 can be displaced by adding excess of soluble headgroup [Ins(1,4,5)P3], membrane activity of PH-PLCdelta1 cannot. PH-PLCdelta1 differs from other phosphoinositide-binding domains in that its membrane insertion does not require that the phosphoinositide binding site be occupied. Significant monolayer insertion remains when the phosphoinositide-binding site is mutated, and PH-PLCdelta1 can insert into monolayers that contain no PtdIns(4,5)P2 at all. Our results suggest a model in which reversible membrane binding of PH-PLCdelta1, mediated by PtdIns(4,5)P2 or other acidic phospholipids, occurs without membrane insertion. Accumulation of the PH domain at the membrane surface enhances the efficiency of insertion, but does not significantly affect its extent, whereas the presence of phosphatidylethanolamine and cholesterol in the lipid mixture promotes the extent of insertion. This is the first report of membrane activity in an isolated PH domain and has implications for understanding the membrane targeting by this common type of domain. PMID- 15755259 TI - Methylglyoxal administration induces diabetes-like microvascular changes and perturbs the healing process of cutaneous wounds. AB - Increased formation of MG (methylglyoxal) and related protein glycation in diabetes has been linked to the development of diabetic vascular complications. Diabetes is also associated with impaired wound healing. In the present study, we investigated if prolonged exposure of rats to MG (50-75 mg/kg of body weight) induced impairment of wound healing and diabetes-like vascular damage. MG treatment arrested growth, increased serum creatinine, induced hypercholesterolaemia (all P < 0.05) and impaired vasodilation (P < 0.01) compared with saline controls. Degenerative changes in cutaneous microvessels with loss of endothelial cells, basement membrane thickening and luminal occlusion were also detected. Acute granulation appeared immature (P < 0.01) and was associated with an impaired infiltration of regenerative cells with reduced proliferative rates (P < 0.01). Immunohistochemical staining indicated the presence of AGEs (advanced glycation end-products) in vascular structures, cutaneous tissue and peripheral nerve fibres. Expression of RAGE (receptor for AGEs) appeared to be increased in the cutaneous vasculature. There were also pro inflammatory and profibrotic responses, including increased IL-1beta (interleukin 1beta) expression in intact epidermis, TNF-alpha (tumour necrosis factor-alpha) in regions of angiogenesis, CTGF (connective tissue growth factor) in medial layers of arteries, and TGF-beta (transforming growth factor-beta) in glomerular tufts, tubular epithelial cells and interstitial endothelial cells. We conclude that exposure to increased MG in vivo is associated with the onset of microvascular damage and other diabetes-like complications within a normoglycaemic context. PMID- 15755260 TI - An ordered sequential mechanism for Factor IX and Factor IXa binding to platelet receptors in the assembly of the Factor X-activating complex. AB - To define the contributions of the Omega-loop of the Gla (gamma-carboxyglutamic acid) domain and the EGF2 (second epidermal growth factor) domain of FIXa (Factor IXa) in the assembly of the FX-activating complex on activated platelets and phospholipid membranes, three recombinant FIXa chimeras were prepared with corresponding residues from the homologous coagulation protein, FVII: (i) Gly4 Gln11 (FIXa7Omegaloop), (ii) Cys88-Cys124 (FIXa7EGF2), and (iii) both Gly4-Gln11 and Cys88-Cys124 (FIXa7Omegaloop7EGF2). All three chimeras were similar to wild type FIXa, as assessed by SDS/PAGE, active-site titration, content of Gla residues, activation rates by FXIa and rates of FXa generation in solution. Titrations of FX or FVIIIa on SFLLRN peptide-activated platelets and on phospholipid vesicles in the presence of FVIIIa revealed normal substrate and cofactor binding to all chimeras. In kinetic assays in the presence of phospholipid vesicles and FVIIIa, compared with wild-type FIXa K(d, app) approximately 4 nM, the FIX7Omegaloop chimera showed a 1.6-fold increase in K(d, app), the FIX7EGF2 chimera had a 7.4-fold increase in K(d, app), and the FIX7Omegaloop7EGF2 chimera showed a 21-fold increase in K(d, app). In kinetic assays and equilibrium platelet-binding assays with activated platelets and FVIIIa, compared with wild-type FIXa (V(max) approximately 5 nM min(-1); K(d, app) approximately 0.5 nM; B(max) approximately 550 sites/platelet; K(d) approximately 0.5 nM), the FIX7Omegaloop chimera displayed 2-fold decreases in V(max) and B(max) and 2-fold increases in K(d, app) and K(d). The FIX7EGF2 chimera displayed 2-fold decreases in V(max) and B(max) and 10-fold increases in K(d, app) and K(d). The FIX7Omegaloop7EGF2 chimera showed non-saturable curves and severely impaired rates of FXa generation, and non-saturable, non-specific, low-level binding to activated platelets. Thus both the Gla domain Omega-loop (Gly4-Gln11) and the EGF2 domain (Cys88-Cys124) are required to mediate the normal assembly of the FX-activating complex on activated platelets and on phospholipid membranes. PMID- 15755261 TI - How effective are the English smoking treatment services in reaching disadvantaged smokers? AB - AIMS: To determine the effectiveness of smoking cessation services in enabling smokers living in disadvantaged areas to access treatment services, and to assess the extent of variations between areas. DESIGN: Observational study of administrative information linked with survey data. SETTING: A representative sample of 19 of the 95 English health areas in 2001. MEASUREMENTS: All England smoking data by deprivation category obtained from the Health Survey of England were used to estimate neighbourhood smoking prevalence rates. Area of residence data from smokers setting a quit date were used to calculate the proportion of smokers in receipt of treatment services in different economic deprivation categories. FINDINGS: In general, treatment services were seeing smokers from the most disadvantaged areas where smoking prevalence rates were highest; 32.3% of all smokers in receipt of treatment services lived in the most disadvantaged quintile of areas compared with 9.6% resident in the most advantaged quintile. An indicator of 'positive discrimination' was calculated for each health authority area to quantify the extent to which the proportion of disadvantaged smokers being treated was greater than the proportion in the local population. This figure ranged from just under 0% to 18%. CONCLUSIONS: National Health Service (NHS) smoking cessation services have been successful in reaching smokers from disadvantaged communities. If improved access to support for smokers living in the poorest communities can be extended, sustained and translated into long-term quitting then smoking cessation services have the potential to make a useful contribution to addressing inequalities in health. PMID- 15755262 TI - The English smoking treatment services: one-year outcomes. AB - AIMS: To assess the impact of English treatment services on CO-validated quit rates at 52-week follow-up, to explore the relationship between service-related characteristics and socio-demographic and behavioural factors with cessation outcomes, and to compare the characteristics of service users lost to follow-up with CO-validated quitters. DESIGN: Observational study of administrative information linked with survey data for 2069 recipients of smoking treatment services who set a quit date between May and November 2002. SETTING: Two contrasting areas of England, Nottingham and North Cumbria, consisting of nine primary care trust (PCT) localities. MEASUREMENTS: Routine monitoring data specified by the Department of Health included information about basic demographic characteristics, postcode of residence from which a deprivation category was identified, nature of intervention, and smoking status at 4-week follow-up. These data were supplemented with information about smoking status at 52 weeks, referral pathways, relapse experiences, number of follow-up contact attempts, socio-economic status and smoking-related behaviours obtained from consenting service recipients by treatment advisers. FINDINGS: One user in seven (14.6%) reported prolonged abstinence and was CO-validated as a successful quitter at 52 weeks. This rose to 17.7% when self-report cases were included. Relapse rates between 4 and 52 weeks were almost identical between the two study areas--75%. Relapse was most likely to occur in the first 6 months following treatment. Users who self-reported quitting at 4 weeks were less likely (13.7%) than those with biochemical verification of smoking status at 4 weeks (25.2%) to be CO-validated quitters at 52 weeks (P = 0.004). Older users (OR 1.023; CI 1.014 1.032), people who smoke mainly for pleasure rather than to cope (OR 1.38; CI 1.02-1.87), and those who were extremely determined (OR 1.58; CI 1.21-2.05) were more likely to be quitters at 52-week follow-up, whereas those with lower socio economic status (OR 0.86; CI 0.78-0.96), who smoked their first cigarette of the day within 5 minutes of waking (OR 0.73; CI 0.55-0.96) or had another smoker in their household (OR 0.65; CI 0.49-0.86) were less likely. In contrast, users lost to follow-up tended to be younger and experienced different referral pathways than CO-validated quitters. Gender was not statistically significantly associated with cessation at 52 weeks and nor were any of the key characteristics of intervention, such as group or one-to-one counselling. CONCLUSIONS: These results obtained from routine services are consistent with those obtained from clinical trials in relation to abstinence at one year. Given that a high proportion of smokers relapsed between 4 weeks and 1 year it is important that future assessments of longer-term outcomes are conducted. However, following-up service users many months after an intervention is expensive, and reasonable estimates of quit rates can be estimated from short-term outcomes, provided that they have been CO-validated. Future studies should monitor outcomes from a selection of services treating different groups of smokers, particularly if more is to be learned about the role of smoking treatment services in reducing inequalities in health. PMID- 15755263 TI - Lessons from the English smoking treatment services. AB - This paper summarizes and discusses the key findings of the evaluation of the English smoking treatment services, which were established in 1999 as part of the English National Health Service. Within 4 years these services existed throughout the country and were working at full capacity, a total of pound 76 million having been spent on them over this period, excluding medication costs. In the fourth year almost 235,000 people attended treatment and set a quit date, and the total budget, including medications, was approximately pound 50 million. At the end of the fourth year the government allocated pound 138 million for the services for the period April 2003-March 2006. The CO-validated 4-week abstinence rate was 53%, the validated 52-week abstinence rate was 15%, and the relapse rate from 4 to 52 weeks was 75%. There was no sex difference in cessation rates at long-term follow-up. The cessation results and relapse rate from weeks 4 to 52 are consistent with results from published studies, including clinical trials. The estimated cost per life-year saved was pound 684 and the figure is even lower if the potential future health care cost savings are taken into account at pound 438 per life-year saved. This compares with the benchmark of pound 20,000 per life year saved, which the National Institute for Clinical Excellence (NICE) is using to recommend new health care interventions in the National Health Service. The services were also succeeding in reaching disadvantaged smokers. However, there have been problems, and other health care systems considering an initiative of this kind should: set national training standards and increase training capacity before launching the services; standardize the provision of pharmaceutical treatments and make them as accessible as possible before launching the services; and give the services at least 5 years of central funding to allow them to become well established. Monitoring is extremely important but should not be so much of a burden that it detracts from developing a quality service and although cessation targets can be helpful, care needs to be taken that they are reasonable and do not promote throughput at the expense of quality. PMID- 15755264 TI - Proceedings of the TransAtlantic-Pacific Dialogues in Acid-related Disorders. May 2004. New Orleans, Louisiana, USA. PMID- 15755265 TI - The appropriate use of proton-pump inhibitor co-therapy with non-steroidal anti inflammatory drugs and cyclo-oxygenase-2 selective inhibitors. PMID- 15755266 TI - Non-steroidal anti-inflammatory drugs and proton-pump inhibitors vs. cyclo oxygenase-2 selective inhibitors in reducing the risk of recurrent ulcer bleeding in patients with arthritis. PMID- 15755267 TI - Optimizing cardiovascular and chemopreventive benefits of aspirin: what role for the proton-pump inhibitors? PMID- 15755268 TI - Endoscopy-negative reflux disease: part of the spectrum of gastro-oesophageal reflux disease [corrected] or a separate disorder? Implications for treatment. PMID- 15755269 TI - Tough-to-treat gastro-oesophageal reflux disease patients: who are they and how to treat? PMID- 15755270 TI - Definitions and classification of dyspepsia: pH, Helicobacter pylori, non steroidal anti-inflammatory drugs--should we include gastro-oesophageal reflux disease? PMID- 15755271 TI - Implications for treatment: pH, Helicobacter pylori or alternative approaches? PMID- 15755272 TI - Managing dyspepsia in regions with a high risk of gastric cancer. PMID- 15755275 TI - Mixed pulmonary fungal infection with Aspergillus fumigatus and Absidia corymbifera in a patient with relapsed acute myeloid leukaemia. PMID- 15755276 TI - Osteonecrosis of the jaw associated with zoledronate therapy in a patient with multiple myeloma. PMID- 15755277 TI - Perforin and lymphohistiocytic proliferative disorders. AB - Perforin is critical for cytotoxicity mediated by granules present in natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). Perforin-deficient mice have impaired cytotoxicity by NK cells and CTLs, resulting in failure to control infections with certain viruses or bacteria. Infection of perforin-deficient mice with lymphocytic choriomeningitis virus results in haemophagocytic lymphohistiocytosis and elevated levels of pro-inflammatory cytokines. Mutations throughout the perforin gene have been identified in patients with familial haemophagocytic lymphohistiocytosis (FHL) type 2. These patients present with fever, hepatosplenomegaly, pancytopenia, have marked elevations of T-helper type 1 and type 2 cytokines, and have impaired NK cell and CTL cytotoxicity. A number of infectious pathogens have been implicated as triggering the onset of disease. Identification of mutations in perforin as the cause of FHL should allow prenatal diagnosis of the disorder. While stem cell transplantation is curative, gene therapy might be effective in the future. PMID- 15755278 TI - Defining stroke risk in children with sickle cell anaemia. AB - Sickle cell anaemia (SCA) is the most common cause of childhood stroke, occurring with the highest frequency before the age of 6 years. Despite the relative frequency of stroke in SCA, few predictors of risk exist. Anaemia, leucocytosis, hypertension, silent infarction, and history of acute chest syndrome are well documented risk factors for ischaemic stroke in SCA. Recent data suggest that other environmental and genetic factors, many unrelated to SCA, influence the development of cerebrovascular disease. Non-invasive assessment of individual stroke risk using transcranial Doppler ultrasonography has provided a means of selecting and prophylactically treating SCA children at highest risk. With the ultimate goal of preventing stroke, the information gained from the studies reviewed here may lead to improved prediction of stroke so that clinical trials to assess risk-based therapy may be carried out on selected children with SCA. PMID- 15755279 TI - c-Flip protein expression in Burkitt's lymphomas is associated with a poor clinical outcome. AB - The World Health Organization Classification of Lymphoid Neoplasms identifies Burkitt's lymphoma/leukaemia (BL) as a single entity, characterized by unique clinical and genetic features that require specific high intensity chemotherapy regimens. Although remarkable successes in the treatment of the disease have been observed, when compared with paediatric patients, adults are less likely to reach stable complete remission. We investigated 32 BL cases, composed in equal part by adults and children that were treated with the French LMB regimen, for factors that may be implicated in chemoresistance. Immunohistochemical detection of procaspase-8, caspase-3a, survivin, p53, CD95, c-Flip and Phospho-RelA (Ser536) was investigated on paraffin-embedded tissues. The expression of c-Flip was found highly related to a poor prognosis, mostly characterized by adults with a chemoresistant disease, resulting in a high death rate within the first year of diagnosis. The 2-year overall survival with c-Flip expression was 24% compared with 93% in the absence of this marker (P = 0.04). All c-Flip-positive BL cases presented a nuclear Phospho-RelA (Ser536) localization, suggesting the presence of an active nuclear factor (NF)-kappa B transcription pathway. These findings show that c-Flip could be a reliable prognostic factor in BL, suggesting new therapeutic approaches that target the NF-kappa B pathway. PMID- 15755280 TI - Concurrent detection of minimal residual disease (MRD) in childhood acute lymphoblastic leukaemia by flow cytometry and real-time PCR. AB - Minimal (i.e. submicroscopic) residual disease (MRD) predicts outcome in childhood acute lymphoblastic leukaemia (ALL). To be used clinically, MRD assays must be reliable and accurate. Two well-established techniques, flow cytometry (FC) and polymerase chain reaction (PCR), can detect leukaemic cells with a sensitivity of 0.01% (10(-4)). We analysed diagnostic samples of 45 ALL-patients (37 B-lineage ALL, eight T-lineage ALL) by four-colour FC and real-time PCR. Leukaemia-associated immunophenotypes, at a sensitivity of MRD detection by FC at the 0.01% level, were identified in 41 cases (91%); antigen-receptor gene rearrangements suitable for MRD detection with a sensitivity of 0.01% or better by PCR were identified in 38 cases (84%). The combined use of FC and PCR allowed MRD monitoring in all 45 patients. In 105 follow-up samples, MRD estimates by both methods were highly concordant, with a deviation factor of <5 by Bland Altman analysis. Importantly, the concordance between FC and PCR was also observed in regenerating bone marrow samples containing high proportions of CD19(+) cells, and in samples studied 24 h after collection. We conclude that both MRD assays yield generally concordant results. Their combined use should enable MRD monitoring in virtually all patients and prevent false-negative results due to clonal evolution or phenotypic shifts. PMID- 15755281 TI - Prediction of individual response to chemotherapy in patients with acute myeloid leukaemia using the chemosensitivity index Ci. AB - As the response to chemotherapy in patients with acute myeloid leukaemia (AML) may still not be accurately determined by known prognostic factors, such as karyotype, the ex vivo chemosensitivity profile may help to predict the individual response. The predictive accuracy of an ex vivo assay should be assessed by correlation of assay results with both response rate and survival. We prospectively investigated the prognostic relevance of pre-therapeutic ex vivo chemosensitivity testing in primary cell cultures from adult AML patients by applying a new evaluation methodology, designated the chemosensitivity index, C(i). This C(i) was designed as a prognostic index by taking the area under the curve as an exact measure of the total dose-response relationship. We found an overall predictive accuracy of 98.2% concerning treatment response, which compares favourably with previously published data ranging from 75% to 92%. Moreover, the C(i) proved to be the strongest prognostic factor for overall survival in a multivariate Cox regression analysis including karyotype grouping and age (P < 0.001), and enabled the evaluation of response to combination therapies and selection of possible treatment alternatives. Our data suggest that ex vivo chemosensitivity testing evaluated by the C(i) could serve as a powerful tool for assay-directed therapy strategies in AML. PMID- 15755282 TI - Incidence and spectrum of non-Hodgkin lymphoma in Chinese migrants to British Columbia. AB - The incidence and spectrum of non-Hodgkin lymphoma (NHL) differ between the Chinese and Caucasian populations. Using population-based registries, we studied the pattern of NHL in Chinese migrants to British Columbia (BC). The records of all NHL cases of Chinese descent diagnosed between 1980 and 1997 were retrieved. Age-standardized incidences were calculated by 5-year intervals in terms of age and calendar years and the relative rates were compared between the migrant, Hong Kong and BC populations. The histological distribution of NHL was compared with 4500 consecutive NHL cases diagnosed in the two populations. A total of 211 cases of migrant NHL were identified, with an age-standardized incidence rate of 7.11 per 100 000 per year, compared with the Hong Kong and BC rates of 7.91 [standardized incidence ratio (SIR) = 0.86, P = 0.01] and 11.88 (SIR = 0.56, P < 0.01). The standardized rates of follicular lymphoma remained low, but the incidence of gastric and nasal natural killer/T lymphomas in migrants were lower than expected. Genetic factors appeared to be stronger than environmental factors in governing the overall incidence of NHL in Chinese. However, certain subtypes of lymphoma may show decreased rates in migrants because of environmental factors. PMID- 15755283 TI - Acute leukaemia in children with Down syndrome: a population-based Nordic study. AB - To determine the epidemiology and outcome of children with Down syndrome (DS) diagnosed with acute leukaemia in the Nordic countries, data registered in the Nordic Society of Paediatric Haematology and Oncology (NOPHO) population-based leukaemia registry were analysed. Of 3494 children with acute leukaemia diagnosed between July 1984 and December 2001, 136 patients (3.9%) with DS were identified. 2.1% of the children with acute lymphoid leukaemia (ALL) and 14.0% of the children with acute myeloid leukaemia (AML) had DS. In ALL, DS patients had similar age and sex distribution and no major differences in blood counts compared with non-DS children. None of the DS patients had T cell leukaemia. Outcome was inferior to that of non-DS children and treatment results did not improve over time. In AML, DS patients showed a significant female predominance and all but one were <5 years old. DS patients with AML had significantly lower platelet and white blood cell counts and two-thirds were type M7 as according to the French-American-British classification. None of the patients <5 years of age had typical AML cytogenetic aberrations. Outcome was far better in the DS group. DS patients treated for AML after 1992 had an excellent outcome (probability of event-free survival, 83 +/- 6%). The high proportion of female DS patients with AML is unexplained. The differing treatment results in AML versus ALL need further evaluation and represent a challenge for the coming years. PMID- 15755284 TI - Methylation status of the p15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia. AB - Aberrant DNA methylation is frequently observed in adults with myelodysplastic syndrome (MDS), and is recognized as a critical event in the disease's pathogenesis and progression. This is the first report to investigate the methylation status of p15 and p16, cell cycle regulatory genes, in children with MDS (n = 9) and juvenile myelomonocytic leukaemia (JMML; n = 18) by using a methylation-specific polymerase chain reaction. The frequency of p15 hypermethylation in paediatric MDS was 78% (7/9), which was comparable to that in adult MDS. In contrast, p15 hypermethylation in JMML was a rare event (17%; 3/18). In JMML, clinical and laboratory characteristics including PTPN11 mutations and aberrant colony formation were not different between the three patients with hypermethylated p15 and the others. Aberrant methylation of p16 was not detected in children with either MDS or JMML. Since p15 and p16 genes were unmethylated in two children with JMML, in whom the disease had progressed with an increased number of blasts, a condition referred to as blastic crisis, we infer that the aberrant methylation of these genes is not responsible for the progression of JMML. The results suggest that demethylating agents may be effective in most children with MDS and a few patients with JMML. PMID- 15755285 TI - Serum levels of inflammatory cytokines at diagnosis correlate to the bcl-6 and CD10 defined germinal centre (GC) phenotype and bcl-2 expression in patients with diffuse large B-cell lymphoma. AB - Circulating inflammatory cytokines have a prognostic impact independent of the information provided by the International Prognostic Index (IPI) in diffuse large B-cell lymphoma (DLBCL). The present study characterized prognostic cytokines in relation to stage-specific B-cell differentiation antigens and bcl-2 protein expression, assessed by immunohistochemistry in de novo DLBCL. Serum levels of interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-alpha) were found to be significantly lower in patients with a germinal centre (GC) phenotype (co expression of bcl-6 and CD10) compared with the non-GC phenotype. IL-6 and TNF alpha levels were significantly elevated in patients expressing bcl-2 protein. Serum levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were not associated with the GC phenotype. On the contrary, both VEGF and bFGF were strongly correlated to bcl-2 expression. In survival analysis, IPI score remained the most important independent prognostic factor. However, IL-6 and VEGF, combined with non-GC phenotype and bcl-2 positivity, respectively, had a similar independent prognostic power as the IPI. In conclusion, our data suggest that inflammatory cytokines are differently distributed in the GC and non-GC phenotypes and correlate to bcl-2 expression. Combining these biomarkers may add to the prognostic information given by clinical variables in the IPI alone. PMID- 15755286 TI - Characterizing CD43 expression in haematogones using multicolor flow cytometric analysis. AB - Haematogones are precursor B cells commonly detected in small numbers in the bone marrow. Morphologically, haematogones can mimic lymphoblasts and are best distinguished using multicolour flow cytometry with antibody combinations. Haematogones show characteristic and reproducible patterns of antigen expression representing the B-cell maturation sequence. CD43 expression, widely seen in haematopoietic elements, has not been well characterized in haematogones. We demonstrate that CD43 is consistently expressed in haematogones in a reproducible pattern similar to that of CD10 when combined with CD20. We propose that in combination with other markers, CD43 can be useful in the identification of haematogones. PMID- 15755287 TI - Use of 18F-FDG positron emission tomography following allogeneic transplantation to guide adoptive immunotherapy with donor lymphocyte infusions. AB - Fluorine-18 fluorodeoxyglucose positron emission tomography ((18)F-FDG PET) provides valuable prognostic information in the management of lymphoma patients. However, the utility of (18)F-FDG PET following allografting is unclear. We analysed the use of (18)F-FDG PET after allogeneic reduced-intensity transplantation (RIT) performed in our institution. Between June 1998 and January 2002, 55 patients underwent RIT for either Hodgkin or non-Hodgkin lymphoma. At least one (18)F-FDG PET scan was performed during the post-transplant period (median five studies) in 15 (27.2%) of these 55 patients. PET scans were performed after re-staging computed tomography (CT) and were categorised depending on (18)F-FDG uptake. The first PET scan was informative in 11 of 15 patients (73%) and influenced the administration of donor lymphocyte infusions (DLI) in nine: leading to earlier DLI administration in two patients, earlier dose escalation in one, withholding of DLI administration in five and dose reduction in one. In addition, subsequent monitoring with (18)F-FDG PET scans documented a graft-versus-lymphoma effect in five patients (median post-DLI follow-up 33 months, range 13-36 months). These preliminary data suggest that (18)F-FDG PET has a role in guiding DLI administration and monitoring the immunotherapeutic effect in patients after allogeneic transplantation. This retrospective pilot study forms the basis for a prospective study to clarify the utility of (18)F-FDG PET/CT in these patients. PMID- 15755289 TI - Changes in platelet count after cardiac surgery can effectively predict the development of pathogenic heparin-dependent antibodies. AB - Cardiopulmonary bypass (CPB) induces the release of platelet factor 4 (PF4) and patients are at risk of heparin-induced thrombocytopenia (HIT). This study was aimed to determine whether an abnormal evolution in platelet count (PC) after CPB is predictive of the development of HIT antibodies. Two abnormal PC patterns were defined: pattern P1, characterized by a decrease in PC following previous correction of thrombocytopenia occurring during CPB, and pattern P2, defined as a persistent low PC in the days following CPB. PC was evaluated for 10 d in 305 consecutive patients before and after CPB. Serotonin release assay (SRA) was carried out between days 8 and 10 to detect pathogenic heparin-dependent antibodies. Moreover, antibodies to heparin-PF4 (H-PF4) complexes were assayed by enzyme-linked immunosorbent assay. PC evolution after CPB was normal in 300 patients although antibodies to H-PF4 were frequently present (53.4%). Changes in PC were abnormal in five patients with pattern P1 (n = 4) or P2 (n = 1). As SRA was positive in four of the five cases, the positive predictive value of abnormal PC pattern for pathogenic HIT antibodies was 80%. Careful follow-up of PC after CPB makes it possible to predict with high specificity (99%) for those patients who develop pathogenic HIT antibodies. PMID- 15755288 TI - The prevalence of the cysteine1584 variant of von Willebrand factor is increased in type 1 von Willebrand disease: co-segregation with increased susceptibility to ADAMTS13 proteolysis but not clinical phenotype. AB - The molecular pathogenesis of type 1 von Willebrand disease (VWD) is uncertain in most patients. We examined 30 type 1 VWD families in the UK Haemophilia Centre Doctors' Organization study. Heterozygosity for Y/C1584 was present in eight of 30 (27%) families and 19 of 76 (25%) individuals with type 1 VWD recruited into the study. Eighteen (95%) of these 19 individuals were blood group O. C1584 did not co-segregate with VWD in four families, and co-segregated in one family; the results were equivocal in three families. In all families increased susceptibility of von Willebrand factor (VWF) to a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS) 13 proteolysis co-segregated with C1584 in affected and unaffected individuals. These data show that C1584, associated with blood group O, is prevalent among patients with type 1 VWD but not necessarily causative of disease and should not be used in isolation to diagnose VWD. Increased susceptibility of C1584 VWF to ADAMTS13 proteolysis may be physiologically significant and increase an individual's risk of bleeding and presenting with VWD. PMID- 15755290 TI - An evaluation of rapid D-dimer assays for the exclusion of deep vein thrombosis. AB - We evaluated the performance of eight d-assays for the exclusion of deep vein thrombosis (DVT); Biopool AutoDimer, Biopool MiniQuant, bioMerieux MDA D-Dimer, VIDAS, Dade Behring D-Dimer Plus, Trinity Biotech AMAX, NycoCard D-dimer and IL Test D-Dimer. The assays were evaluated both as stand-alone tests, and in combination with pretest probability (PTP). D-dimer assays and PTP assessment were performed on 410 patients presenting to the emergency department with suspected acute DVT. DVT was diagnosed in 76 of 410 patients (18.5%) by compression ultrasound or other imaging techniques, as required. Receiver operator characteristics analysis established optimum cut-off values and these were compared with manufacturer's cut-off values where provided. As stand-alone tests, the assays varied immensely regarding cut-off value, negative predictive value (NPV 93-100%) and specificity (0-67%). At least one patient with confirmed DVT had a low d-dimer level by each method: to achieve 100% sensitivity it would be necessary to reduce cut-off values to levels below clinical usefulness. When low d-dimer was used in combination with PTP, six of eight methods achieved > or =98% NPV, with a diagnosis of DVT excluded in 16-44% of patients without the requirement for diagnostic imaging. The highly variable diagnostic performance of these d-dimer assays means that some assays are unsuitable for certain diagnostic strategies. However, our data suggest that the combination of sensitive D-dimer assays with an assessment of PTP may be used to exclude a diagnosis of DVT. PMID- 15755291 TI - Interleukin 4, interleukin 6 and interleukin 10 polymorphisms in children with acute and chronic immune thrombocytopenic purpura. AB - Immune thrombocytopenic purpura (ITP) is an immune-mediated disorder. We investigated the polymorphisms of interleukin (IL)-4 intron 3, IL-6 (-572 G/C), and IL-10 (-627 C/A) in 50 children with acute ITP, 30 children with chronic TIP, and 100 healthy individuals. There were significant differences in the RP1/RP2 genotype proportion (P = 0.04) and the RP2 allelic frequency (P = 0.03) of IL-4 intron 3 and the A/C genotype proportion (P = 0.01) of IL-10 (-627) between children with chronic ITP and controls. This finding suggests that the IL-4 intron 3 and IL-10 (-627) polymorphisms contribute to the susceptibility of developing childhood chronic ITP. PMID- 15755292 TI - Requirement for myeloid growth factors in the differentiation of acute promyelocytic leukaemia. AB - It is well known that the differentiation of acute promyelocytic leukaemia (APL) cells by all-trans-retinoic acid (ATRA) may be enhanced by myeloid growth factors, but the requirement for growth factors in this process is unclear. Our previous studies in multiple myeloma and non-APL acute myeloid leukaemia demonstrated that lineage-specific growth factors are required for the maximal activity of many pharmacologic differentiating agents in vitro. Thus, we studied whether the differentiation of APL is similarly dependent on growth factors. We found that the myeloid growth factors granulocyte colony-stimulating factor or granulocyte-macrophage colony-stimulating factor markedly increased the differentiation of NB4 cells or APL blasts from clinical samples treated with ATRA, arsenic trioxide (ATO), or bryostatin-1 as evidenced by the enhanced expression of myeloid surface antigens and the inhibition of clonogenic growth. Furthermore, myeloid growth factors were necessary for the differentiation of APL cells since the activity of each pharmacologic agent could be blocked by specific growth factor-neutralizing antibodies. Each differentiating agent was active only at concentrations that inhibited cell cycling, suggesting that this property is also required for differentiation. These data demonstrate that both pharmacologic differentiating agents and myeloid growth factors are required, but neither sufficient, for the differentiation of APL cells. The combined use of pharmacologic differentiating agents and growth factors may improve the clinical efficacy of differentiation therapy in APL. PMID- 15755293 TI - Activated T-lymphocytes with myelosuppressive properties in patients with chronic idiopathic neutropenia. AB - To characterize the cellular components responsible for the impaired granulopoiesis in chronic idiopathic neutropenia (CIN), we investigated the origin of the proapoptotic cytokine producing cells in the bone marrow (BM) microenvironment of CIN patients. We found that the interferon gamma (IFN gamma) and/or Fas-ligand expressing cells in patient BM mononuclear cells and long-term BM culture stroma cells were the CD3(+) T-lymphocytes but not the CD14(+) monocytes/macrophages. The percentage of activated T-lymphocytes was increased in patients' BM as indicated by the proportions of human leucocyte antigen (HLA) DR(+), CD25(+), CD38(+), CD69(+) and Fas(+) cells within the CD3(+) fraction. Intracellular IFN gamma expression was higher in the BM than peripheral blood of the patients and was associated with increased BM T-lymphocyte numbers. In crossover experiments, patient CD3(+) T-lymphocytes conferred autologous and allogeneic haemopoietic progenitor cell colony inhibition. Patients' T-cell receptor repertoire and polymerase chain reaction analysis did not reveal any clonal T-lymphocyte expansion, suggesting the absence of a direct, antigen-driven recognition of CD34(+) myeloid progenitor cells by patient T-lymphocytes. We conclude that CIN patients have increased number of activated T-lymphocytes in the BM, probably in the setting of a localized polyclonal immune reaction and that these cells confer an inhibitory effect on myelopoiesis through myelosuppressive cytokines including Fas-ligand and IFN gamma. PMID- 15755294 TI - Overexpression of the Doppel protein in acute myeloid leukaemias and myelodysplastic syndromes. AB - We investigated the expression patterns and distribution of Doppel (Dpl), the product of the prion-like gene PRND, in the leukaemic cell lines HL-60 and K562 and in bone marrow cells from 44 patients with acute myeloid leukaemia (AML) and 63 patients with myelodysplastic syndrome (MDS). Whereas normal samples were negative or showed very weak expression that was restricted to CD34(+) cells, Dpl was detected in both cell lines and in most AML and MDS cases by immunocytochemistry and Western blotting. Quantitative reverse transcription polymerase chain reaction revealed variable mRNA levels in almost all AML and MDS cases, but barely detectable levels in normal bone marrow. These differences were confirmed by in situ hybridization. PRND expression was higher in advanced compared with early MDS (P = 0.01), but Dpl levels did not predict disease progression. In AML there was no correlation between Dpl levels and clinical or laboratory findings. In conclusion, this is the first time that the expression of PRND has been demonstrated in human bone marrow. The molecular mechanism of the observed overexpression is unknown; however, the differential Dpl distribution in AML and MDS versus healthy subjects makes it a possible leukaemia-associated antigen that could be useful for diagnostic and therapeutic purposes. PMID- 15755295 TI - A simple inhibitor screen is more sensitive than a Bethesda assay in monitoring for the development of inhibitors in haemophilia A and B. PMID- 15755296 TI - Pulmonary hypertension and thalidomide therapy in multiple myeloma. PMID- 15755298 TI - Editorial. PMID- 15755299 TI - Annotation: PANDAS: a model for human autoimmune disease. AB - BACKGROUND: Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcus infections (PANDAS) is a recently recognized syndrome in which pre adolescent children have abrupt onsets of tics and/or obsessive-compulsive symptoms, a recurring and remitting course of illness temporally related to streptococcal infections, and associated neurologic findings including adventitious movements, hyperactivity and emotional lability. METHODS: Inspired by observations of similar symptoms in children with Sydenham's chorea, a search was undertaken for clinical and laboratory evidence in support of the new syndrome. RESULTS: Consistent and predictable clinical findings have been described in a large case series. Magnetic resonance imaging has supported the postulated pathobiology of the syndrome with evidence of inflammatory changes in basal ganglia. Antibasal ganglia antibodies have been found in some acute cases, mimicking streptococcal antigen epitopes. CONCLUSIONS: While PANDAS remains a controversial diagnostic concept, it has stimulated new research endeavors into the possible links between bacterial pathogens, autoimmune reactions, and neuropsychiatric symptoms. PMID- 15755300 TI - When more is not better: the role of cumulative risk in child behavior outcomes. AB - BACKGROUND: Cumulative risk research has established the deleterious effects of co-occurring risk factors on child behavior outcomes. However, extant literature has not addressed potential differential effects of cumulative risk at different points in development and has left open questions about whether a threshold model or a linear risk model better describes the impact of cumulative risk on behavior outcomes. The current study examined the impact of cumulative risk factors (i.e., child maltreatment, inter-parental violence, family disruption, low socioeconomic status, and high parental stress) in early and middle childhood on child behavior outcomes in adolescence. METHODS: Using data from an ongoing longitudinal study of at-risk urban children (N=171), the cumulative effects of these five risk factors across early and middle childhood were investigated. RESULTS: The findings support the cumulative risk hypothesis that the number of risks in early childhood predicts behavior problems in adolescence. Evidence for a linear but not a threshold model of cumulative risk was found; the more risks present, the worse the child outcome. Moreover, the presence of multiple risks in early childhood continues to explain variations in predicting adolescent behavior outcomes even after including the effects of risk in middle childhood. CONCLUSIONS: The results support the need for comprehensive prevention and early intervention efforts with high-risk children, such that there does not appear to be a point beyond which services for children are hopeless, and that every risk factor we can reduce matters. PMID- 15755301 TI - Are maternal smoking and stress during pregnancy related to ADHD symptoms in children? AB - BACKGROUND: There are some indications that maternal lifestyle during pregnancy (smoking and stress) contributes to symptoms of ADHD in children. We prospectively studied whether prenatal exposure to maternal smoking and/or stress is associated with ADHD symptoms and diagnostic criteria (according to DSM-IV) in 7-year-olds. METHODS: Nulliparous Scandinavian women were consecutively recruited at their first prenatal health care visit and assessments of smoking and stress were collected at gestational weeks 10, 12, 20, 28, 32, and 36. Children were followed up at 7 years old. We obtained full data for 72% of the sample: ADHD symptoms were rated by 74% of mothers (n=290) and 96% of eligible teachers (n=208). Attrition analyses showed no differences on key variables between participants and non-participants at follow-up. RESULTS: Results of multiple regression analyses showed prenatal exposure to smoking (beta=.16, p<.01) and stress (beta=.18, p<.01) were independently associated with later symptoms of ADHD. Results of logistic regression analyses showed that fulfillment of the diagnostic criteria for ADHD was related to exposure to prenatal stress (beta=.68, p<.01) especially in boys. The results were not confounded by sociodemographic factors or birth outcomes. CONCLUSIONS: This study provides evidence that prenatal exposure to stress and smoking is independently associated with later symptoms of ADHD in human children, particularly for boys. Because stress and smoking are relatively common during pregnancy, and yet preventable, these results are of public health significance. PMID- 15755302 TI - The adolescent outcome of hyperactive girls: self-report of psychosocial status. AB - BACKGROUND: The aim of the study was to clarify the developmental risk associated with hyperactive behaviour in girls in a longitudinal epidemiological design. METHODS: This was investigated in a follow-up study of girls who were identified by parent and teacher ratings in a large community survey of 6- and 7-year-olds as showing pervasive hyperactivity or conduct problems or the comorbid mixture of both problems or neither problem. They were later investigated, at the age of 14 to 16 years, with a detailed self-report interview technique. RESULTS: Hyperactivity was a risk factor for later development, even allowing for the coexistence of conduct problems. Hyperactivity predicted academic problems and interpersonal relationship problems. Relationships with parents, by contrast, were not portrayed to be as problematic as relationships with peers and the opposite sex. Their psychological, social and occupational functioning was objectively rated to be more deviant and their self-report showed them to be more ambivalent about their future. There was a trend for hyperactivity to be self reported as a risk for the development of continuing symptomatology but neither hyperactivity nor conduct problems were self-reported to be a risk for antisocial behaviour, substance misuse or low self-esteem in adolescence. However, they were at risk for the development of state anxiety. CONCLUSIONS: The results suggested girls' pattern of functioning may differ from that of boys because girls self report a more pervasive range of social dysfunction than that previously reported in boys. PMID- 15755303 TI - The importance of parenting in the development of disorganized attachment: evidence from a preventive intervention study in adoptive families. AB - BACKGROUND: As infant disorganized attachment is a serious risk factor for later child psychopathology, it is important to examine whether attachment disorganization can be prevented or reduced. METHOD: In a randomized intervention study involving 130 families with 6-month-old adopted infants, two attachment based intervention programs were tested. In the first program, mothers were provided a personal book, and in the second program mothers received the same personal book and three home-based sessions of video feedback. The third group did not receive intervention (control group). RESULTS: The intervention with video feedback and the personal book resulted in enhanced maternal sensitive responsiveness (d=.65). Children of mothers who received this intervention were less likely to be classified as disorganized attached at the age of 12 months (d=.46), and received lower scores on the rating scale for disorganization than children in the control group (d=.62). In the book-only intervention group children showed lower disorganization ratings compared to the control group, but no effect on the number of infants with disorganized attachment classifications was found. CONCLUSION: Our short-term preventive intervention program with video feedback and a book lowered the rate of disorganized attachment. The effectiveness of our intervention documents the importance of parenting in the development of infant attachment disorganization. PMID- 15755304 TI - Prevalence of pervasive developmental disorders in children and adolescents with mental retardation. AB - BACKGROUND: Insight into the prevalence of pervasive developmental disorder (PDD) in children and adolescents with mental retardation (MR) is known to be of clinical importance. However, estimating this prevalence is complicated. The literature reports prevalence rates ranging from 3% through 50%. This variation seems to be related to the concepts of PDD under study, the instruments used, and the studied populations. The present study aimed to estimate a reliable prevalence rate of PDD. METHODS: A total population-based screening with the PDD MRS and the ABC (n=825) was followed by further assessment of children and adolescents at high risk for PDD according to these instruments, and for controls, with the ADI-R, ADOS-G and a DSM-IV-TR classification (n=188). RESULTS: The instruments lead to different prevalence rates that range from 7.8% to 19.8%. The differences in the estimated prevalence rates are related to the concept of PDD and the instruments they represent. The DSM-IV-TR prevalence (16.7%) seems to be the most reliable and well-founded estimate, since this prevalence rate is based on information from multiple informants and multiple time periods. CONCLUSIONS: The reported prevalence rates provide policy makers with an up-to date and more substantiated guideline for the allocation of resources for children and adolescents with MR and PDD. The height of the prevalence should alert professionals that PDD is widespread in the population with MR. PMID- 15755305 TI - The Carter Neurocognitive Assessment for children with severely compromised expressive language and motor skills. AB - In this paper, different means of assessing cognitive development in children with severe impairments in both their expressive language and their motor skills are reviewed. A range of techniques are considered, including traditional cognitive tests and behavioral and physiological measures, but these techniques are generally impractical and minimally informative when it comes to assessing children with both motor and speech impairments. Electrophysiological measures show some promise for the future, but are currently inadequate for wide-ranging cognitive assessment. Development of the Carter Neurocognitive Assessment (CNA) is described. The CNA is appropriate for use in clinical and research settings and was designed to minimalize the impact of severely impaired motor skills and expressive language on performance. The CNA is intended to itemize and quantify a range of skills reflecting a cognitive level up to approximately 18 to 24 months in four areas: Social Awareness, Visual Attention, Auditory Comprehension and Vocal Communication. The use of the CNA to assess the performance and developmental growth of eight children with Holoprosencephaly (HPE), a midline developmental brain malformation, is described. The CNA is a useful tool for the assessment of children with severely compromised motor and verbal skills and has provided a more positive view of the cognitive potential of children with severe handicaps, such as the sample of children with HPE, than that presented in the past. PMID- 15755306 TI - Serial order reconstruction in Down syndrome: evidence for a selective deficit in verbal short-term memory. AB - BACKGROUND: Individuals with Down syndrome consistently perform less well than appropriately matched comparison groups on tests of verbal short-term memory, despite performing relatively well on non-verbal short-term memory tasks. However, it is not clear whether these findings constitute evidence for a selective deficit in verbal short-term memory, or whether they instead reflect the influence of non-central factors such as speech difficulties or poor number knowledge. METHODS: Twenty-six individuals with Down syndrome and 32 typically developing children were tested on a digit reconstruction task in which participants were presented with auditory digit sequences and responded by pressing the corresponding digits on a touch-screen in the correct serial order. Background measures were performance on a closely matched visuo-spatial reconstruction task, reaction time on a simple digit identification task, receptive vocabulary age and non-verbal ability (Raven's matrices). Participants were also tested on a conventional digit recall task. RESULTS: All four background measures accounted for significant individual variation in digit reconstruction performance, but there remained a significant effect of group that reflected relatively poor performance of individuals with Down syndrome. Hierarchical regression showed that group membership accounted for unique variation in both digit reconstruction and recall performance, even after all group differences on background measures had been accounted for. CONCLUSIONS: The results provide strong evidence that Down syndrome is associated with a selective deficit in verbal short-term memory, and a deficit in verbal serial order memory in particular. Implications for the language difficulties associated with Down syndrome are discussed. PMID- 15755307 TI - Non-verbal cognitive development and language impairment. AB - BACKGROUND: Specific language impairment (SLI) is currently partly defined by the presence of non-verbal IQ scores in the normal range. However, not only is there a debate concerning where 'normal thresholds' should be, but increasing information about the presence of processing deficits in SLI have led some researchers to question the use of IQ criteria in clinical diagnosis. In particular, little is known about the longitudinal and developmental patterns of cognitive performance in this population. METHOD: Data from a long-term follow-up study of SLI was examined in 82 children defined at original participation as having SLI who had IQ measurements at 7, 8, 11 and 14 years. RESULTS: Analyses revealed a significant fall between 7 and 14 years of over 20 IQ points. This fall took place mainly between 8 and 11 years but was still continuing between 11 and 14 years. Further investigation revealed different groups of children showing different developmental patterns in IQ, even after controlling for baseline measurement. These groups also showed significantly different language outcomes at 14 years. Analyses controlling for IQ at 7 were also performed that suggested a dynamic process between language and cognitive development. CONCLUSIONS: These findings appear to be in agreement with a model of impairment that views 'Residual Normality' as unlikely (Karmiloff-Smith, 1998; Thomas & Karmiloff Smith, 2002). The implications are therefore discussed in relation to the dynamic development of systems along with the possible cognitive mechanisms (such as working memory) that might interact with language to create an SLI profile. PMID- 15755308 TI - Deafness to fear in boys with psychopathic tendencies. AB - The processing of the emotional signals of others is fundamental for normal socialization and interaction. Reduced responsiveness to the expressions of sadness and fear has been implicated in the development of psychopathy (Blair, 1995). The current study investigates the ability of boys with psychopathic tendencies to process auditory affect information. Boys with psychopathic tendencies and a comparison group, as defined by the Antisocial Process Screening Device (APSD: Frick & Hare, 2001), were presented with neutral words spoken with intonations conveying happiness, disgust, anger, sadness and fear and were asked to identify the emotion of the speaker based on prosody. The boys with psychopathic tendencies presented with a selective impairment for the recognition of fearful vocal affect. These results are interpreted with reference to amygdala dysfunction and components of the Integrated Emotion Systems model. PMID- 15755309 TI - Veterinary drug usage and antimicrobial resistance in bacteria of animal origin. AB - In the production of food animals, large amounts of antimicrobial agents are used for therapy and prophylaxis of bacterial infections and in feed to promote growth. There are large variations in the amounts of antimicrobial agents used to produce the same amount of meat among the different European countries, which leaves room for considerable reductions in some countries. The emergence of resistant bacteria and resistance genes due to the use of antimicrobial agents are well documented. In Denmark it has been possible to reduce the usage of antimicrobial agents for food animals significantly and in general decreases in resistance have followed. Guidelines for prudent use of antimicrobial agents may help to slow down the selection for resistance and should be based on knowledge regarding the normal susceptibility patterns of the causative agents and take into account the potential problems for human health. Current knowledge regarding the occurrence of antimicrobial resistance in food animals, the quantitative impact of the use of different antimicrobial agents on selection of resistance and the most appropriate treatment regimes to limit the development of resistance is incomplete. Programmes monitoring the occurrence and development of resistance and consumption of antimicrobial agents are strongly desirable, as is research into the most appropriate ways to use antimicrobial agents in veterinary medicine. PMID- 15755310 TI - Enhanced transcription of contractile 5-hydroxytryptamine 2A receptors via extracellular signal-regulated kinase 1/2 after organ culture of rat mesenteric artery. AB - 5-Hydroxytryptamine (5-HT) has been found to elicit enhanced contractile effects in some vascular disorders. The present study was designed to examine if vascular 5-HT2A receptors are up-regulated during organ culture and if the extracellular signal-regulated protein kinase 1/2 (ERK1/2) pathways are involved. Compared with fresh rat mesenteric artery ring segments, the contractile responses to 5-HT were significantly increased in the segments cultured for 6, 24 or 48 hr (P<0.05, P<0.01, P<0.01, respectively). The 5-HT-induced contraction occurred via 5-HT2A receptors, since the selective 5-HT2A antagonist ketanserin blocked the 5-HT induced contraction in the fresh segments with a pA2 value 9.5 (slope was 0.98 with 95% confidence intervals from 0.8 to 1.1). A similar result was obtained in the segments cultured for 24 hr with a pA2 value of 9.43 (slope=0.91 and 95% confidence intervals between 0.45 to 2.3). In addition, the enhanced 5-HT2A receptor contraction occurred with a significant increase of 5-HT2A receptor mRNA (P<0.05). Organ culture of the mesenteric artery was found to activate ERK1/2 already within 1 and 3 hr. It is likely that the ERK1/2 pathways were involved as a initial switch, since the selective ERK1/2 pathway inhibitor SB386023 abolished both up-regulation of 5-HT2A mRNA transcription and the enhanced contractile response to 5-HT. These data reveal a role of ERK1/2 in up-regulation of 5-HT2A receptors and suggest a possibility to inhibit the enhanced responses to 5-HT by inhibition of the ERK1/2 pathway. PMID- 15755311 TI - Evaluation of a drug-drug interaction alert structure through the retrospective analysis of statins-macrolides co-prescriptions. AB - The aim of this work was the evaluation of the existent drug interaction alert structure in Spain, which is based on yellow cards notifications and circulation of drug alert letters, through the retrospective analysis of CYP3A-metabolized statins and macrolides co-prescriptions in the Spanish province of Badajoz between May and September 2001. The period of study was planned to include the release of 2 drug alert letters released by the Spanish Drug Agency in June and July, addressed to all healthcare professionals, which warned against the concomitant prescription of statins and inhibitors of their metabolism, e.g. macrolides antibacterials. 4,600,764 prescriptions were examined, 664 of which corresponded to combinations of statins and macrolides. Although a decrease was detected in the number of these co-prescriptions throughout the study, 80 of these corresponding to 67 patients were still being prescribed in September, after the warnings by the Spanish Drug Agency had been released. 431 physicians prescribed these drugs simultaneously, with 22.9% of them having more than one patient at potential risk. Doctors working at rural healthcare centres or not directly attached to any healthcare facility were more prone to prescribe unsafe coprescriptions than those working at urban health centre. This study shows that the present drug alert system is not fully efficient when facing a situation like the one retrospectively reviewed in this study, in which a prompt action, in this case termination of potentially hazardous coprescriptions, was required. New systems developed to improve prescribing, including a new method based on personal contact between Drug Surveillance Centres and general practitioners, are discussed. PMID- 15755312 TI - Metal binding ability of metallothionein-3 expressed in Escherichia coli. AB - Metallothionein-3/growth inhibitory factor is a brain-specific member of the metallothionein gene family, and impairs the survival and neurite formation of cultured neurons. Metallothionein-3 can bind heavy metals such as Zn, Cu, or Cd almost in the same way as other metallothionein family. However, its biological function as growth inhibitory factor apparently distinguishes metallothionein-3 from other metallothioneins. To better understanding of the relationship between the growth inhibitory activity of metallothionein-3 and metals bound to metallothionein-3, the metal-binding ability of metallothionein-3 was analyzed in comparison with those of metallothionein-1 and -2. The metal-binding ability of metallothionein-3 was evaluated by pH titration and 5-5' dithiobis (2 nitrobenzoic) acid (DTNB) analysis as compared with those of the other metallothioneins. The affinity of metal ions for metallothionein-3 was indicated as follows, Cu>Cd>Zn, same as metallothionein-1 and -2. However, the affinity of metallothionein-3 to Cu was much higher than that of metallothionein-1 and -2. The strong affinity to Cu of metallothionein-3 might be related to its growth inhibitory activity. PMID- 15755313 TI - 2,3-Dimercapto-1-propanol does not alter the porphobilinogen synthase inhibition but decreases the mercury content in liver and kidney of suckling rats exposed to HgCl2. AB - Heavy metals have received great attention as environmental pollutants mainly because once introduced in the biological cycle they are incorporated in the food chain. Especially the mercury toxicity due to a diversity of effects caused by different chemical species should be emphasized. Heavy metal intoxication has been treated with chelating agents such as 2,3-dimercapto-1-propanol (BAL). However, the efficacy of this treatment is questionable due to the lack of specific effect on the toxic metal. The present study examined the effects of HgCl2 exposure (five doses of 5.0 mg/kg between ages 8 to 12 days) on physiological parameters, on porphobilinogen synthase activity, and on mercury content in liver, kidneys and brain from suckling rats. The effect of BAL (one dose of 12.5-75 mg/kg) applied 24 hr after mercury intoxication on these parameters was also investigated. The results demonstrate that HgCl2 intoxication induced a decrease of corporal weight gain as well as brain weight and an increase in renal weight. The inhibition of porphobilinogen synthase from liver and kidney, is still significant and was not modified by subsequent BAL treatment. However, BAL altered two effects induced by mercury: increase in death percentage and decrease in mercury contents in liver and kidney. The increase of mortality induced by mercury was not promoted by metal redistribution to brain nor by the increase of porphobilinogen synthase inhibition induced by metal. More investigations are necessary to determine if the different effects of BAL on intoxication by metals are possibly related to other tissues and/or if the probable metal-chelating complex formed is more toxic than the metal itself. PMID- 15755314 TI - Xenobiotics and the glucocorticoid receptor: additive antagonistic effects on tyrosine aminotransferase activity in rat hepatoma cells. AB - Methylsulfonyl-PCBs (MeSO2-PCBs) and some fungicides were studied for their functional effects on the glucocorticoid signal transduction in the Reuber rat hepatoma H-II-E-C3 cell line. 4-Substituted MeSO2-PCBs, tolylfluanid and ketoconazole displayed antagonistic effects on dexamethasone-induced tyrosine aminotransferase specific activity (IC50 ranging from 0.7-5.1 microM), but no agonist activity. These substances also had affinity to the mouse glucocorticoid receptor in competition binding studies, indicating that the inhibition of the middle cerebral artery occlusion-activity is indeed mediated by receptor binding. Thus, substances with a structural resemblance with a methyl sulfonyl group, such as the fungicide tolylfluanid, may inhibit glucocorticoid receptor-regulated gene transcription. In co-exposure experiments with three substances, multivariate modelling showed that the inhibitory effect of 4-MeSO2-2,5,6,2',4' pentachlorobiphenyl (4-MeSO2-CB91), 4-MeSO2-2,3,6,2',4',5'-hexachlorobiphenyl (4 MeSO2-CB149) and tolylfluanid on tyrosine aminotransferase activity was close to additive. Thus, co-exposure to such different chemicals as persistent organic pollutants and pesticides may affect cells additively. Chemical interference with the glucocorticoid hormone system therefore deserves further attention in vivo. PMID- 15755315 TI - The broad-spectrum cation channel blocker pinokalant (LOE 908 MS) reduces brain infarct volume in rats: a temperature-controlled histological study. AB - Activation of cation channels conducting Ca2+, Na+ and K+ is involved in the pathogenesis of infarction in experimental focal cerebral ischaemia. Pinokalant (LOE 908 MS) is a novel broad-spectrum inhibitor of several subtypes of such channels and has previously been shown to improve the metabolic and electrophysiologic status of the ischemic penumbra and to reduce lesion size on magnetic resonance images in the acute phase following middle cerebral artery occlusion in rats. The purpose of the present study was to investigate whether these beneficial effects of pinokalant are translated into permanent neuroprotection in terms of a reduction in infarct size one week after middle cerebral artery occlusion in rats. Halothane-anaesthetized male Wistar rats subjected to permanent distal middle cerebral artery occlusion were randomly assigned to one of two treatment groups: 1) Control (vehicle intravenous loading dose followed by infusion); 2) Pinokalant (0.5 mg/kg intravenous loading dose followed by infusion of 1.25 mg/kg/hr). Infusions started 30 min. after middle cerebral artery occlusion and were continued for 24 hr. Body temperature and mean arterial blood pressure were monitored by telemetry during this period and the spontaneous temperature after course in control rats established in other experiments was imitated. Seven days later histological brain sections were prepared and the infarct volumes measured. Body temperature did not differ between the groups. Mean arterial blood pressure was slightly higher in the pinokalant group. Pinokalant treatment significantly reduced cortical infarct volume from 33.8+/-15.8 mm3 to 24.5+/-13.1 mm3 (control group versus pinokalant group, P=0.017, t-test). Taking the effective drug plasma concentration established in other experiments into account revealed that in rats with plasma concentrations within the therapeutic interval, infarct volumes were further reduced to 17.9+/-7.5 mm3 (P<0.005). PMID- 15755316 TI - The structure of (S)-(-)-4-(2-benzimidazolyl)-4-hydroxybutyric acid formed during mammalian metabolism of the fungicide 2-(2-furyl)benzimidazole. AB - During metabolism of the fungicide 2-(2-furyl)benzimidazole (FB) in mammals, an optically active compound, metabolite A, was isolated. This compound, a principal metabolite in the biological degradation of FB, was isolated from the urine of horse and dog. When studied by IR, NMR, MS, CD and electrophoretic techniques, the metabolite proved to be the zwitterionic form of (S)-(-)-4-(2-benzimidazolyl) 4-hydroxybutyric acid. The synthesized optical active compound was in every respect identical with metabolite A. The suggested structure has been completely verified by the current investigation of its crystal structure, based on single crystal diffraction data collected at the synchrotron MAX II in Lund, Sweden. In an earlier determination, a related compound, 3-benzimidazolylpropionic acid (Ercan et al. 1996) was suggested to be non-zwitterionic. The present studies have shown that the latter compound too is zwitterionic and that the conclusions presented were apparently based on some erroneously proposed hydrogen positions. PMID- 15755317 TI - 2,3-Dimercaptopropanol, 2,3-dimercaptopropane-1-sulfonic acid and meso-2,3 dimercaptosuccinic acid acute administration diferentially change biochemical parameters in mice. PMID- 15755318 TI - Automatic detection of false annotations via binary property clustering. AB - BACKGROUND: Computational protein annotation methods occasionally introduce errors. False-positive (FP) errors are annotations that are mistakenly associated with a protein. Such false annotations introduce errors that may spread into databases through similarity with other proteins. Generally, methods used to minimize the chance for FPs result in decreased sensitivity or low throughput. We present a novel protein-clustering method that enables automatic separation of FP from true hits. The method quantifies the biological similarity between pairs of proteins by examining each protein's annotations, and then proceeds by clustering sets of proteins that received similar annotation into biological groups. RESULTS: Using a test set of all PROSITE signatures that are marked as FPs, we show that the method successfully separates FPs in 69% of the 327 test cases supplied by PROSITE. Furthermore, we constructed an extensive random FP simulation test and show a high degree of success in detecting FP, indicating that the method is not specifically tuned for PROSITE and performs well on larger scales. We also suggest some means of predicting in which cases this approach would be successful. CONCLUSION: Automatic detection of FPs may greatly facilitate the manual validation process and increase annotation sensitivity. With the increasing number of automatic annotations, the tendency of biological properties to be clustered, once a biological similarity measure is introduced, may become exceedingly helpful in the development of such automatic methods. PMID- 15755319 TI - Increased capsaicin receptor TRPV1 in skin nerve fibres and related vanilloid receptors TRPV3 and TRPV4 in keratinocytes in human breast pain. AB - BACKGROUND: Breast pain and tenderness affects 70% of women at some time. These symptoms have been attributed to stretching of the nerves with increase in breast size, but tissue mechanisms are poorly understood. METHODS: Eighteen patients (n = 12 breast reduction and n = 6 breast reconstruction) were recruited and assessed for breast pain by clinical questionnaire. Breast skin biopsies from each patient were examined using immunohistological methods with specific antibodies to the capsaicin receptor TRPV1, related vanilloid thermoreceptors TRPV3 and TRPV4, and nerve growth factor (NGF). RESULTS: TRPV1-positive intra epidermal nerve fibres were significantly increased in patients with breast pain and tenderness (TRPV1 fibres / mm epidermis, median [range] - no pain group, n = 8, 0.69 [0-1.27]; pain group, n = 10, 2.15 [0.77-4.38]; p = 0.0009). Nerve Growth Factor, which up-regulates TRPV1 and induces nerve sprouting, was present basal keratinocytes: some breast pain specimens also showed NGF staining in supra-basal keratinocytes. TRPV4-immunoreactive fibres were present in sub-epidermis but not significantly changed in painful breast tissue. Both TRPV3 and TRPV4 were significantly increased in keratinocytes in breast pain tissues; TRPV3, median [range] - no pain group, n = 6, 0.75 [0-2]; pain group, n = 11, 2 123, p = 0.008; TRPV4, median [range] - no pain group, n = 6, [0-1]; pain group, n = 11, 1 [0.5 2], p = 0.014). CONCLUSION: Increased TRPV1 intra-epidermal nerve fibres could represent collateral sprouts, or re-innervation following nerve stretch and damage by polymodal nociceptors. Selective TRPV1-blockers may provide new therapy in breast pain. The role of TRPV3 and TRPV4 changes in keratinocytes deserve further study. PMID- 15755320 TI - Secondary structure in the target as a confounding factor in synthetic oligomer microarray design. AB - BACKGROUND: Secondary structure in the target is a property not usually considered in software applications for design of optimal custom oligonucleotide probes. It is frequently assumed that eliminating self-complementarity, or screening for secondary structure in the probe, is sufficient to avoid interference with hybridization by stable secondary structures in the probe binding site. Prediction and thermodynamic analysis of secondary structure formation in a genome-wide set of transcripts from Brucella suis 1330 demonstrates that the properties of the target molecule have the potential to strongly influence the rate and extent of hybridization between transcript and tethered oligonucleotide probe in a microarray experiment. RESULTS: Despite the relatively high hybridization temperatures and 1M monovalent salt imposed in the modeling process to approximate hybridization conditions used in the laboratory, we find that parts of the target molecules are likely to be inaccessible to intermolecular hybridization due to the formation of stable intramolecular secondary structure. For example, at 65 degrees C, 28 +/- 7% of the average cDNA target sequence is predicted to be inaccessible to hybridization. We also analyzed the specific binding sites of a set of 70mer probes previously designed for Brucella using a freely available oligo design software package. 21 +/- 13% of the nucleotides in each probe binding site are within a double-stranded structure in over half of the folds predicted for the cDNA target at 65 degrees C. The intramolecular structures formed are more stable and extensive when an RNA target is modeled rather than cDNA. When random shearing of the target is modeled for fragments of 200, 100 and 50 nt, an overall destabilization of secondary structure is predicted, but shearing does not eliminate secondary structure. CONCLUSION: Secondary structure in the target is pervasive, and a significant fraction of the target is found in double stranded conformations even at high temperature. Stable structure in the target has the potential to interfere with hybridization and should be a factor in interpretation of microarray results, as well as an explicit criterion in array design. Inclusion of this property in an oligonucleotide design procedure would change the definition of an optimal oligonucleotide significantly. PMID- 15755321 TI - Design of the BiRmingham Early Detection In untREated psyChosis Trial (REDIRECT): cluster randomised controlled trial of general practitioner education in detection of first episode psychosis [ISRCTN87898421]. AB - BACKGROUND: Treatment delay in first episode psychosis is common. As general practitioners are the first point of contact for many individuals with first episode psychosis, they are well placed to detect the early symptoms and make urgent referrals to specialist secondary care services. However, early psychosis is often difficult to detect. The primary objective of the Redirect trial is to estimate whether an educational intervention targeted at general practitioners increases the general practitioner referral rate of young people with first episode psychosis to Early Intervention Services. METHODS/DESIGN: This paper describes the design of a stratified-cluster randomised controlled trial of an educational intervention on first episode psychosis in primary care. The primary outcome is the number of general practitioner referrals of young people with first episode psychosis to Early Intervention Services. Secondary outcomes are duration of untreated psychosis, time to recovery, use of the Mental Health Act, and general practitioner consultation rate. Young people with first episode psychosis referred to Early Intervention Services will be recruited over a two year period from 1 March 2004. Seventy-eight out of 89 eligible general practices were recruited. The educational intervention has been implemented and evaluated by general practitioners. The education was well received and considered relevant to clinical practice by the general practitioners. DISCUSSION: The results suggest that the recruitment strategy and implementation of the educational intervention are feasible and acceptable in a primary care setting. The Redirect trial will provide robust information about the efficacy of an evidence-based complex educational intervention targeted at general practitioners on referral rates of young people with first episode psychosis to Early Intervention Services. PMID- 15755322 TI - Assessing the capacity of the health services research community in Australia and New Zealand. AB - BACKGROUND: In order to profile the health services research community in Australia and New Zealand and describe its capacity, a web-based survey was administered to members of the Health Services Research Association of Australia and New Zealand (HSRAANZ) and delegates of the HSRAANZ's Third Health Services Research and Policy Conference. RESULTS: Responses were received from 191 individuals (68%). The responses of the 165 (86%) who conducted or managed health services research indicated that the health services research community in Australia and New Zealand is characterised by highly qualified professionals who have come to health services research via a range of academic and professional routes (including clinical backgrounds), the majority of whom are women aged between 35 and 54 who have mid- to senior- level appointments. They are primarily employed in universities and, to a lesser extent, government departments and health services. Although most are employed in full time positions, many are only able to devote part of their time to health services research, often juggling this with other professional roles. They rely heavily on external funding, as only half have core funding from their employing institution and around one third have employment contracts of one year or less. Many view issues around building the capacity of the health services research community and addressing funding deficits as crucial if health services research is to be translated into policy and practice. Despite the difficulties they face, most are positive about the support and advice available from peers in their work settings, and many are actively contributing to knowledge through academic and other written outputs. CONCLUSION: If health services research is to achieve its potential in Australia and New Zealand, policy-makers and funders must take the concerns of the health services research community seriously, foster its development, and contribute to maximising its capacity through a sustainable approach to funding. There is a clear need for a strategic approach, where the health services research community collaborates with competitive granting bodies and government departments to define and fund a research agenda that balances priority-driven and investigator driven research and which provides support for training and career development. PMID- 15755323 TI - Persistent demographic differences in colorectal cancer screening utilization despite Medicare reimbursement. AB - BACKGROUND: Colorectal cancer screening is widely recommended, but often under utilized. In addition, significant demographic differences in screening utilization exist. Insurance coverage may be one factor influencing utilization of colorectal cancer screening tests. METHODS: We conducted a retrospective analysis of claims for outpatient services for Washington state Medicare beneficiaries in calendar year 2000. We determined the proportion of beneficiaries utilizing screening fecal occult blood tests, flexible sigmoidoscopy, colonoscopy, or double contrast barium enema in the overall population and various demographic subgroups. Multiple logistic regression analysis was used to determine the relative odds of screening in different demographic groups. RESULTS: Approximately 9.2% of beneficiaries had fecal occult blood tests, 7.2% had any colonoscopy, flexible sigmoidoscopy, or barium enema (invasive) colon tests, and 3.5% had invasive tests for screening indications. Colonoscopy accounted for 41% of all invasive tests for screening indications. Women were more likely to receive fecal occult blood test screening (OR 1.18; 95%CI 1.15, 1.21) and less likely to receive invasive tests for screening indications than men (OR 0.80, 95%CI 0.77, 0.83). Whites were more likely than other racial groups to receive any type of screening. Rural residents were more likely than urban residents to have fecal occult blood tests (OR 1.20, 95%CI 1.17, 1.23) but less likely to receive invasive tests for screening indications (OR 0.89; 95%CI 0.85, 0.93). CONCLUSION: Reported use of fecal occult blood testing remains modest. Overall use of the more invasive tests for screening indications remains essentially unchanged, but there has been a shift toward increased use of screening colonoscopy. Significant demographic differences in screening utilization persist despite consistent insurance coverage. PMID- 15755325 TI - Patient-reported outcome and quality of life instruments database (PROQOLID): frequently asked questions. AB - The exponential development of Patient-Reported Outcomes (PRO) measures in clinical research has led to the creation of the Patient-Reported Outcome and Quality of Life Instruments Database (PROQOLID) to facilitate the selection process of PRO measures in clinical research. The project was initiated by Mapi Research Trust in Lyon, France. Initially called QOLID (Quality of Life Instruments Database), the project's purpose was to provide all those involved in health care evaluation with a comprehensive and unique source of information on PRO and HRQOL measures available through the Internet.PROQOLID currently describes more than 470 PRO instruments in a structured format. It is available in two levels, non-subscribers and subscribers, at http://www.proqolid.org. The first level is free of charge and contains 14 categories of basic useful information on the instruments (e.g. author, objective, original language, list of existing translations, etc.). The second level provides significantly more information about the instruments. It includes review copies of over 350 original instruments, 120 user manuals and 350 translations. Most are available in PDF format. This level is only accessible to annual subscribers. PROQOLID is updated in close collaboration with the instruments' authors on a regular basis. Fifty or more new instruments are added to the database annually.Today, all of the major pharmaceutical companies, prestigious institutions (such as the FDA, the NIH's National Cancer Institute, the U.S. Veterans Administration), dozens of universities, public institutions and researchers subscribe to PROQOLID on a yearly basis. More than 800 users per day routinely visit the database. PMID- 15755324 TI - Interrupted or continuous slowly absorbable sutures - design of a multi-centre randomised trial to evaluate abdominal closure techniques INSECT-trial [ISRCTN24023541]. AB - BACKGROUND: The closure of the abdomen after median laparotomy is still a matter of debate among surgeons. Further well designed and performed randomised controlled trials determining the optimal method of abdominal fascial closure are needed. DESIGN: This is a three armed, multi-centre, intra-operatively randomised, controlled, patient blinded trial. Over 20 surgical departments will enrol 600 patients who are planned for an elective primary abdominal operation. The objective of this study is to compare the frequency of abdominal incisional hernias between two continuous suture techniques with different, slowly absorbable monofilament materials and an interrupted suture using an absorbable braided suture material at one year postoperatively. CONCLUSION: This trial will answer the question whether the continuous abdominal wall closure with a slowly absorbable material with longitudinal elasticity is superior to the continuous suture with a material lacking elasticity and to interrupted sutures with braided thread. PMID- 15755327 TI - Activation of p53, inhibition of telomerase activity and induction of estrogen receptor beta are associated with the anti-growth effects of combination of ovarian hormones and retinoids in immortalized human mammary epithelial cells. AB - BACKGROUND: A full-term pregnancy has been associated with reduced risk for developing breast cancer. In rodent models, the protective effect of pregnancy can be mimicked with a defined regimen of estrogen and progesterone combination (E/P). However, the effects of pregnancy levels of E/P in humans and their underlying mechanisms are not fully understood. In this report, we investigated the growth inhibitory effects of pregnancy levels of E/P and both natural and synthetic retinoids in an immortalized human mammary epithelial cell line, 76N TERT cell line. RESULTS: We observed that cell growth was modestly inhibited by E/P, 9-cis-retinoic acid (9-cis RA) or all-trans-retinoic acid (ATRA), and strongly inhibited by N-(4-hydroxyphenyl) retinamide (HPR). The growth inhibitory effects of retinoids were further increased in the presence of E/P, suggesting their effects are additive. In addition, our results showed that both E/P and retinoid treatments resulted in increased RARE and p53 gene activity. We further demonstrated that p53 and p21 protein expression were induced following the E/P and retinoid treatments. Furthermore, we demonstrated that while the telomerase activity was moderately inhibited by E/P, 9-cis RA and ATRA, it was almost completely abolished by HPR treatment. These inhibitions on telomerase activity by retinoids were potentiated by co-treatment with E/P, and correlated well with their observed growth inhibitory effects. Finally, this study provides the first evidence that estrogen receptor beta is up-regulated in response to E/P and retinoid treatments. CONCLUSION: Taken together, our studies show that part of the anti-growth effects of E/P and retinoids is p53 dependent, and involve activation of p53 and subsequent induction of p21 expression. Inhibition of telomerase activity and up-regulation of estrogen receptor beta are also associated with the E/P- and retinoid-mediated growth inhibition. Our studies also demonstrate that the potency of retinoids on cell growth inhibition may be increased through combination of estrogen and progesterone treatment. PMID- 15755326 TI - Antibiotic activity of telithromycin and comparators against bacterial pathogens isolated from 3,043 patients with acute exacerbation of chronic bronchitis. AB - BACKGROUND: Antimicrobial therapy is considered an important component in the medical management of most patients with acute exacerbation of chronic bronchitis (AECB). The three predominant bacterial species isolated are nontypeable Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. Staphylococcus aureus is also frequently isolated while atypical bacteria are thought to cause up to 10% of exacerbations. Antibacterial resistance is increasing worldwide and little surveillance data exist concerning pathogens isolated from patients with AECB. METHODS: This study examines the prevalence of antibacterial resistance in isolates obtained from patients with clinically diagnosed AECB. A total of 3043 isolates were obtained from 85 centres in 29 countries, between 1999-2003, and were tested against the new ketolide telithromycin and a panel of commonly used antibiotics. RESULTS AND DISCUSSION: Of the S. pneumoniae isolates, 99.9% were susceptible to telithromycin, but only 71% were susceptible to erythromycin and 75.3% to penicillin. Of the H. influenzae isolates, 99.6% were susceptible to telithromycin. 11.7% of these isolates produced beta-lactamase. Almost 10% of S. pneumoniae were multidrug resistant; 99.0% of these isolates were susceptible to telithromycin. Telithromycin also demonstrated good in vitro activity against M. catarrhalis (MIC90 = 0.12 mg/L) and was the most active compound against methicillin susceptible S. aureus (98.9% susceptible). CONCLUSION: Telithromycin demonstrated similar or better activity against the bacterial species investigated than the other agents, with the most complete coverage overall. These species are the predominant causative bacterial pathogens in AECB and thus the spectrum of activity of telithromycin makes it a potential alternative for the empirical treatment of AECB. PMID- 15755328 TI - Patient preferences for notification of normal laboratory test results: a report from the ASIPS Collaborative. AB - BACKGROUND: Many medical errors occur during the laboratory testing process, including lost test results. Patient inquiry concerning results often represents the final safety net for locating lost results. This qualitative study sought to identify, from a patient perspective, specific preferences and factors that influence the process of communicating normal (negative) laboratory test results to patients. METHODS: We conducted 30-minute guided interviews with 20 adult patients. Patients were recruited from two practice-based research networks in Colorado that were participating in a medical errors study. A semi-structured interview elicited the participant's experience with and preference for laboratory test result notification. Quantitative descriptive statistics were generated for demographic and preference data. Qualitative results were analyzed by a team of experienced qualitative researchers using multiple styles of qualitative analyses, including a template approach and an editing approach. RESULTS: Ninety percent of participants wanted to be notified of all tests results. Important issues related to notification included privacy, responsive and interactive feedback, convenience, timeliness, and provision of details. Telephone notification was preferred, followed by regular mail. Electronic notification was perceived as uncomfortable because it was not secure. While 65% preferred being notified by a provider, participants acknowledge that this may be impractical; thus, they wanted to be notified by someone knowledgeable enough to answer questions. Participants do not normally discuss their preferences for test result notification with their providers. CONCLUSION: Privacy, responsive and interactive feedback, convenience, and timeliness with detailed information may be critical for patient satisfaction and for improving patient safety, and are features that may be incorporated into emerging communication channels. PMID- 15755329 TI - Differences between men with screening-detected versus clinically diagnosed prostate cancers in the USA. AB - BACKGROUND: The advent of prostate specific antigen (PSA) testing in the United States of America (USA) has led to a dramatic increase in the incidence of prostate cancer in the United States as well as the number of men undergoing aggressive treatment with radical prostatectomy and radiation therapy. We compared patient characteristics and treatment selection between American men with screening-detected versus clinically diagnosed prostate cancers. METHODS: We evaluated 3,173 men with prostate cancer in the USA. Surveys and medical records provided information on demographics, socioeconomic status, comorbidities, symptoms, tumor characteristics, and treatment. We classified men presenting with symptoms of advanced cancer - bone pain, weight loss, or hematuria - as "clinically diagnosed"; asymptomatic men and those with only lower urinary tract symptoms were considered "screening-detected." We used multivariate analyses to determine whether screening predicted receiving aggressive treatment for a clinically localized cancer. RESULTS: We classified 11% of cancers as being clinically diagnosed. Men with screening-detected cancers were more often non Hispanic white (77% vs. 65%, P < 0.01), younger (36% < 65 years vs. 25%, P or= high school vs. 67%, P < 0.01), healthier (18% excellent health vs. 10%, P < 0.01), and diagnosed with localized disease (90% vs. 75%, P < 0.01). Men with screening-detected localized cancers more often underwent aggressive treatment, 76% vs. 70%, P = 0.05. CONCLUSION: Most cancers were detected by screening in this American cohort. Appropriately, younger, healthier men were more likely to be diagnosed by screening. Minority status and lower socio-economic status appeared to be screening barriers. Screening detected earlier-stage cancers and was associated with receiving aggressive treatment. PMID- 15755364 TI - Management of morbid obesity: surgical options. PMID- 15755365 TI - Management of morbid obesity: before and after surgery: the team approach to management. PMID- 15755366 TI - Management of morbid obesity: bariatric surgery in context. PMID- 15755330 TI - Cost savings associated with improving appropriate and reducing inappropriate preventive care: cost-consequences analysis. AB - BACKGROUND: Outreach facilitation has been proven successful in improving the adoption of clinical preventive care guidelines in primary care practice. The net costs and savings of delivering such an intensive intervention need to be understood. We wanted to estimate the proportion of a facilitation intervention cost that is offset and the potential for savings by reducing inappropriate screening tests and increasing appropriate screening tests in 22 intervention primary care practices affecting a population of 90,283 patients. METHODS: A cost consequences analysis of one successful outreach facilitation intervention was done, taking into account the estimated cost savings to the health system of reducing five inappropriate tests and increasing seven appropriate tests. Multiple data sources were used to calculate costs and cost savings to the government. The cost of the intervention and costs of performing appropriate testing were calculated. Costs averted were calculated by multiplying the number of tests not performed as a result of the intervention. Further downstream cost savings were determined by calculating the direct costs associated with the number of false positive test follow-ups avoided. Treatment costs averted as a result of increasing appropriate testing were similarly calculated. RESULTS: The total cost of the intervention over 12 months was $238,388 and the cost of increasing the delivery of appropriate care was $192,912 for a total cost of $431,300. The savings from reduction in inappropriate testing were $148,568 and from avoiding treatment costs as a result of appropriate testing were $455,464 for a total savings of $604,032. On a yearly basis the net cost saving to the government is $191,733 per year (2003 Can dollars) equating to $3,687 per physician or $63,911 per facilitator, an estimated return on intervention investment and delivery of appropriate preventive care of 40%. CONCLUSION: Outreach facilitation is more expensive but more effective than other attempts to modify primary care practice and all of its costs can be offset through the reduction of inappropriate testing and increasing appropriate testing. Our calculations are based on conservative assumptions. The potential for savings is likely considerably higher. PMID- 15755367 TI - Monsters in our midst? Media and the power of language. PMID- 15755368 TI - Vitamin E not helpful, perhaps harmful. PMID- 15755369 TI - Acupuncture effective for osteoarthritis of the knee. PMID- 15755371 TI - Ten years after bariatric surgery: weight loss sustained, diabetes and hypertension reversed. PMID- 15755370 TI - Use CCBs as last resort in treatment of hypertension. PMID- 15755372 TI - Guidelines for the treatment of chronic stable angina. PMID- 15755373 TI - Consumer-directed health care: One step forward, two steps back? PMID- 15755374 TI - What can we expect from nonoperative treatment options for shoulder pain? PMID- 15755375 TI - Puzzling palmar papules and pits. PMID- 15755376 TI - Somatization: Diagnosing it sooner through emotion-focused interviewing. PMID- 15755377 TI - Preterm labor: Diagnostic and therapeutic options are not all alike. PMID- 15755378 TI - Managing diarrhea in infants. PMID- 15755379 TI - Community-acquired bacterial respiratory tract infections. PMID- 15755380 TI - Non-consented IUD placement reported by Mexican immigrants: A caution for caregivers in the US? PMID- 15755381 TI - Clinical inquiries. Should liver enzymes be checked in a patient taking niacin? PMID- 15755382 TI - Clinical inquiries. How useful is high-sensitivity CRP as a risk factor for coronary artery disease? PMID- 15755383 TI - Clinical inquiries. How should we follow up a positive screen for anemia in a 1 year old? PMID- 15755384 TI - Clinical inquiries. What is the best treatment for analgesic rebound headaches? PMID- 15755385 TI - Clinical inquiries. Is sputum evaluation useful for patients with community acquired pneumonia? PMID- 15755386 TI - Clinical inquiries. What is the best regimen for newly diagnosed hypertension? PMID- 15755387 TI - [Polymorphisms in cystathionine beta-synthase and methylenetetrahydrofolate reductase genes as risk factors for cerebral vascular disease]. AB - BACKGROUND AND OBJECTIVE: High plasma total homocysteine (tHcy), low dietary intake of folate and other B vitamins, and genetic polymorphisms related to the metabolism of homocysteine may interactively contribute to the risk of cerebral vascular disease (CVD). We explored interrelations between total homocysteine levels and mutations in genes for the two key enzymes in methionine-homocysteine metabolism. PATIENTS AND METHOD: We analyzed two polymorphisms, C677T in the MTHFR gene and 844ins68 in the CBS gene. We assessed their association with fasting homocysteine in 64 patients with CVD, and in 159 controls. RESULTS: No differences in CBS and MTHFR genotype frequencies between cases and controls were found (C677T p = 0.87 and 844ins68 p = 0.63), nor was a particular CBS and MTHFR genotype associated with an elevated risk of CVD. None of the genotypes defined by the CBS and MTHFR variants studied showed an association with elevated fasting homocysteine concentrations (C677T p = 0.07 and 844ins68 p = 0.47). CONCLUSIONS: We did not find any indication that genetic variation in the CBS and MTHFR genes are associated with homocysteine-related risk of CVD, hence needing further investigation. The contributions to total plasma homocysteine levels of the common mutations of genes coding for the enzymes controlling homocysteine metabolism are modest. PMID- 15755388 TI - [Predictive factors for readmission in heart failure patients]. AB - BACKGROUND AND OBJECTIVE: Heart failure (HF) is a prevalent and increasing disease and represents one of the main causes of hospital admission. It is associated with an important morbi-mortality and a high rate of readmission. The objective of this study was to know the clinical characteristics of admitted patients with HF and to detect any valuable prognosis factors. PATIENTS AND METHOD: Prospective study of admitted HF patients between May'99 and May'00. Readmission rate was evaluated six months later. RESULTS: 204 patients were included with an average age of 78 (9.9) years. 66% were women. Diabetes mellitus (DM) (36.4%) and chronic obstructive pulmonary disease (COPD) (23.4%) were the most outstanding associated pathologies. Ischemic heart disease (IHD) was the most frequent etiology in 33.4% cases. Mean time of admission was 10 days. 34% patients had systolic dysfunction (SD). Men with IHD presented a higher rate of SD (p < 0.001). Mortality was 12.4%, especially in COPD patients (p < 0.011). IECAs were prescribed in 71.2% in the SD group. Readmission rate at 6 months was about 43%. Patients with renal failure (p < 0.04) and those with a more impaired functional class (p < 0.02) displayed a higher readmission rate. CONCLUSIONS: Several clinical factors determine the morbi-mortality and prognosis including an older age, associated comorbility, type of cardiopathy and presence of systolic dysfunction. All these factors are detected at the time of hospital admission. PMID- 15755389 TI - [Intramuscular route for the administration of the anti-flu vaccine in patients receiving oral anticoagulation therapy]. AB - BACKGROUND AND OBJECTIVE: Comparison of safety and effectiveness of the intramuscular (IM) (deltoid) vs subcutaneous (s.c.) administration of the flu vaccine (FV) in patients on oral anticoagulation therapy. PATIENTS AND METHOD: It was a phase IV, simple blind, 2-parallel groups, randomized trial developed in an urban primary care setting. We included patients taking oral anticoagulation therapy without FV contraindications. The IM administration of the FV in the experimental group was compared with a SC administration in the control group. RESULTS: 59 patients were included. The two groups were comparable at the beginning of the study. INR was not modified following s.c. (p = 0.38) or i.m. (p = 0.49) administration. No systemic side effects were observed. More cutaneous lesions were observed in the S.C. group (71.4%) when compared with the i.m. group (25.8%; p < 0.0001). For the remaining variables, we observed a tendency towards more reported pain in the SC group (35.7 vs 22.6%) and a larger brachial diameter (42 vs 29%) without significant differences. 3.2% of patients in the i.m. group and 7.1% in the s.c. group developed influenza symptoms without significant differences. CONCLUSIONS: Even though the results must be interpreted with caution, currently there appears to be no apparent contraindication for the i.m. administration of the FV. When it is administered subcutaneously, it tends to cause more side effects. PMID- 15755390 TI - [Lipid profile in the hyperacute phase of ischemic stroke allows to decide cholesterol-lowering therapy]. AB - BACKGROUND AND OBJECTIVE: We aimed to determine whether measuring the hyperacute lipid profile helps to start a cholesterol-lowering therapy soon after an ischemic stroke. PATIENTS AND METHOD: 72 ischemic stroke patients underwent lipid determinations at three times (hyperacute phase or arrival, second day and the third month). RESULTS: Total cholesterol, cLDL, cHDL and triglyceride mean values were significantly higher (p < 0.05) at arrival compared to the second day (212 vs 189; 131 vs 120; 52 vs 44 and 132 vs 115, respectively). In the third month, mean values were similar to those of the hyperacute phase (197, 131, 48 and 143, respectively). Considering the updated cholesterol management guidelines, we should treat 55% of patients diagnosed at the hyperacute phase but only 37% of patients diagnosed at the second day. CONCLUSION: Hyperacute lipid profile determination helps to determine those patients who need a cholesterol-lowering therapy without delay. PMID- 15755391 TI - [Risk factors of ischemic stroke. Who was the sinner, the patient or his/her parents?]. PMID- 15755392 TI - [Carbon monoxide: an invisible and silent lethal killer]. PMID- 15755393 TI - [Relevance of treatment compliance in heart failure]. AB - It is recognized that the irregularity in therapeutic compliance is one of the main unstabilizing factors leading to hospitalization in patients with heart failure (HF). In this review, we specifically deal with the Pharmacological Therapeutic Adherence (PTA) in HF patients, and especially with the adherence problems related to those drugs which have been shown to improve the prognosis of the disease. The impact of a deficient PTA (DPTA) jeopardizes the efficiency of the new neurohormonal inhibitor drugs which have proven benefit in wide and expensive clinical trials. It is necessary to have more information about PTA, to develop skills and methods to identify noncompliant patients and to practically improve those actions which have shown some positive effect on DPTA. Some DPTA related problems owe to inappropriate therapeutic schemes, adverse effects, social deprivation, scarce interaction with the physician and an inadequate health education. Uninformed patients use to believe that they must take the medicines only when they feel sick and have symptoms, yet they believe that drugs can be withdrawn when they feel better. Different pharmacological groups may have different adherence problems in HF. It is estimated that those interventions aimed at improving adherence are useful for the reduction of health costs are they are likely more effective than the effects caused by choosing a given drug. The choosing method to assess PTA in practice with reliability is the counting of pills combined with a compliance survey. Time spent to improve adherence not only can improve it but also it can diminish the total time spent by the physician in the follow-up of these patients. PMID- 15755394 TI - [Effort intolerance in a 24-year-old male]. PMID- 15755395 TI - [Indications and advantages of laparoscopic splenectomy in patients infected by HIV]. PMID- 15755396 TI - [The impact of a simple educational intervention in diagnosis delay among hospitalized patients with pulmonary tuberculosis]. PMID- 15755398 TI - [Hyperbaric tolerance test. Conceptual remarks]. PMID- 15755400 TI - [Comments about diagnosis and treatment of pneumococcal meningitis]. PMID- 15755401 TI - [Collision tumour. adenocarcinoma and synchronous MALT gastric lymphoma]. PMID- 15755403 TI - [Lymphangioleiomyomatosis in a postmenopausal woman under estrogen therapy]. PMID- 15755402 TI - [Dichloropropane poisoning: report of 2 cases]. PMID- 15755404 TI - Geriatrics: contemporary and future concerns. PMID- 15755405 TI - The elderly at risk for periodontitis and systemic diseases. AB - The increasing population of older subjects with dental care needs will become a major challenge to our society and its care providers. To manage the health care needs of the elderly, a coordination between medical and dental care providers will become necessary. From the dental perspective, it is important to develop skills in the risk assessment of older patients. Such risk assessment of older subjects should take an approach that is holistic and focused on the reduction of the infectious burden and the improvement of self-efficacy. PMID- 15755406 TI - Dental caries in older adults. AB - Dental caries is one of the most significant health problems facing older adults. More than half of the elderly who are dentate are affected with either coronal or root caries, and caries is the primary cause of tooth loss in this population. New materials and techniques are emerging to help with geriatric preventive and restorative needs, but ongoing vigilance for caries will be required in this population, which is experiencing increased longevity and tooth retention. PMID- 15755407 TI - Salivary hypofunction and xerostomia: diagnosis and treatment. AB - Salivary gland hypofunction and complaints of xerostomia are common in elderly patients, irrespective of their living situation. Medication use is frequently related to dry mouth symptoms and reductions in salivary flow rates. Patients with reduced salivary flow are at increased risk for caries, oral fungal infections, swallowing problems, and diminished or altered taste. Oral health care providers should institute aggressive preventive measures and recommend palliative care for patients with significant reduction in salivary gland function. The systemic agents pilocarpine and cevimeline may help selected patients. Selective use of fluoride-releasing restorative materials and conservative treatment plans are recommended for this patient group. PMID- 15755408 TI - Derangement, osteoarthritis, and rheumatoid arthritis of the temporomandibular joint: implications, diagnosis, and management. AB - Temporomandibular joint (TMJ) dysfunction is often believed to bea young person's malady. However, geriatric patients also present with clinical findings of TMJ clicking, locking, crepitation, limited opening, and pain. With our aging population and the high prevalence of rheumatic and musculoskeletal diseases in the elderly, it is important to understand the etiopathogenesis, clinical presentation, and management of derangement, rheumatoid arthritis, and osteoarthritis of the TMJ. Although arthritis of the TMJ usually causes only mild to-moderate dysfunction in older patients, they present challenges related to medication use and comorbidity. This article presents the most recent understanding and therapeutic protocols for patient diagnosis and management. PMID- 15755409 TI - Orofacial pain and sensory disorders in the elderly. AB - Many orofacial pain conditions occur in the elderly. Specifically,this article reviews the prevalence of general and orofacial-related pain in the elderly. The authors also describe and discuss the likely disorders and diseases that produce facial pain and burning pain in the mouth. They do not cover jaw joint pain, oral sores, or ulceration-induced pain, as these conditions are better discussed in the context of arthritis and oral pathologies of the mouth. The authors discuss oral motor disorders, myogenous pain, vascular pain, headaches, trigeminal neuralgia, trigeminal neuropathic dis-ease, postherpetic neuralgia, burning mouth syndrome, and occlusal dysesthesia. PMID- 15755410 TI - Underserved elderly issues in the United States: burdens of oral and medical health care. AB - The elderly represent approximately 12.4% of the general population (2000 Census), yet their health care expenditure and consumption represent 14% of the total (2003). Although 10% of the elderly had no medical insurance in 2000, 78% had no dental insurance. Elderly Americans' burden of medical care overuse is worsened by their out-of-pocket expenses for oral health, because this is usually not a covered benefit. In underserved communities, the management of the oral health and dental care needs of older Americans approaches negligence. PMID- 15755411 TI - Interaction with other health team members in caring for elderly patients. AB - Comprehensive health care of the geriatric patient requires thoughtful communication and coordination of services. Unlike young adults, older patients are likely to be frail, have multiple chronic conditions, and experience disability. Hence they are more likely to depend on others for help and to see a variety of health professionals at different sites. This complexity demands that health care professionals consider their care not in isolation, but as part of a team. They must ensure that other members of the team are kept informed and are consulted as appropriate to ensure safe and effective care. Accomplishing this goal requires being acquainted with the usual care providers, the necessary information for sharing, and the most effective communication methods within the team. PMID- 15755412 TI - Cognitive function, aging, and ethical decisions: recognizing change. AB - As the population ages, dental and other health care providers will be working with more older adults (and their family members) with changing cognitive status than ever before in history. The intent of this article is to review common cognitive changes in older adults that will undoubtedly be seen in dental practices. Knowledge of the common signs and symptoms of age-related cognitive changes provides a basis on which to identify individuals with undiagnosed cognitive,changes. This article reviews the relationship between cognitive function, aging, and dementia (specifically, mild cognitive impairment and Alzheimer's disease), the role of the dental team in recognizing these conditions, and issues related to obtaining in-formed consent from cognitively impaired patients. PMID- 15755413 TI - Medication use and prescribing considerations for elderly patients. AB - Older adults consume more medications than any other segment of the population. Increasing lifespan means that more people will live into old age, frequently with disabilities and conditions man-aged by medications. Age-associated physiologic changes, medication use patterns, and adverse drug effects and interactions place the older adult at high risk for medication-related problems. Older adults living in institutions, those with complex medical problems,and those who do not adhere to medication regimens are at highest risk for negative health outcomes from medication mishaps. Dentists must be able to identify older adults who are susceptible to adverse drug events and to recognize which medications are most likely to precipitate problems. PMID- 15755414 TI - Caring for elderly long-term care patients: oral health-related concerns and issues. AB - Much work is needed to resolve the many issues of prevention in the complex environment of long-term care facilities and to provide effective curative care for individuals, no matter how frail, who could benefit from comprehensive dental services. PMID- 15755415 TI - Oral diagnostics for the geriatric populations: current status and future prospects. AB - Because it is a noninvasive technique, there is growing interest in replacing blood with oral-based methods of diagnostics. Oral diagnostics may be used for diagnosis and therapeutic drug monitoring of both oral diseases (eg, caries, periodontal disease,oral lesions, oral cancer) and systemic diseases (eg, infectious diseases, including HIV and AIDS, autoimmune diseases, cancer,and endocrine disorders). The authors address both existing techniques and oral-based diagnostics that will be applicable to the aging population in the future. They also highlight those techniques that are uniquely suited to point-of-care applications. PMID- 15755416 TI - Osteoporosis: diagnostic testing, interpretation, and correlations with oral health--implications for dentistry. AB - Osteoporosis affects over 10 million Americans, including over 2 million men. Dentistry is in a position to work alongside medicine to assist the population in preventing and coping with this condition. This article provides an overview of osteoporosis, including its causes, assessment, prevention, and treatment. Various pharmacologic agents are reviewed, including bisphosphonates, selective estrogen receptor modulators, calcitonin, anabolic agents, and fluoride. The article also discusses the dentist's role in prevention, risk assessment, diagnosis, nutrition counseling, and evaluating the impact of the disease and its treatment on dental management and treatment planning. As dental implants become more in demand, the prognosis for and potential complications of implant therapy in patients with osteoporosis are a matter of intense interest. PMID- 15755418 TI - Leishmania chagasi/infantum: further investigations on Leishmania tropisms in atypical cutaneous and visceral leishmaniasis foci in Central America. AB - In Central America, apparently genetically identical Leishmania chagasi/infantum parasites cause cutaneous (CL) and visceral leishmaniasis (VL), the latter being more frequent in young children. The present study investigated if there were pathology-related differences in virulence between Honduran CL and VL strains using Mediterranean L. infantum strains as a reference. Macrophage infectivity and serum sensitivity, properties thought to be associated with virulence, were similar between CL and VL strains from both regions. Attention focused on the genome organisation of genes for two candidate virulence factors: Leishmania mitogen activated protein kinase (LMPK) and cysteine proteinase b (Cpb). Interestingly, the Mediterranean strains exhibited restriction enzyme polymorphisms associated with tropism for both LMPK and Cpb genes whereas no differences were observed for the Honduran strains. We also report relative genetic homogeneity of the Honduran strains as compared to the Mediterranean strains and discuss it in terms of the probable origin for the Central American L. chagasi/infantum. PMID- 15755417 TI - Impact of vitamin E or selenium deficiency on nematode-induced alterations in murine intestinal function. AB - The effects of deficiencies in the antioxidant nutrients, vitamin E and selenium, on the host response to gastrointestinal nematode infection are unknown. The aim of the study was to determine the effect of antioxidant deficiencies on nematode induced alterations in intestinal function in mice. BALB/c mice were fed control diets or diets deficient in selenium or vitamin E and the response to a secondary challenge inoculation with Heligmosomoides polygyrus was determined. Egg and worm counts were assessed to determine host resistance. Sections of jejunum were mounted in Ussing chambers to measure changes in permeability, absorption, and secretion, or suspended in organ baths to determine smooth muscle contraction. Both selenium and vitamin E deficient diets reduced resistance to helminth infection. Vitamin E, but not selenium, deficiency prevented nematode-induced decreases in glucose absorption and hyper-contractility of smooth muscle. Thus, vitamin E status is an important factor in the physiological response to intestinal nematode infection and may contribute to antioxidant-dependent protective mechanisms in the small intestine. PMID- 15755419 TI - Clonorchis sinensis: molecular cloning and functional expression of novel cytosolic malate dehydrogenase. AB - The NAD-dependent cytosolic malate dehydrogenase (cMDH, EC 1.1.1.37) plays a pivotal role in the malate-aspartate shuttle pathway that operates in a metabolic coordination between cytosol and mitochondria, and thus is crucial for the survival and pathogenicity of the parasite. In the high throughput sequencing of the cDNA library constructed from the adult stage of Clonorchis sinensis, a cDNA clone containing 1152bp insert was identified to encode a putative peptide of 329 amino acids possessing more than 50% amino acid sequence identities with the cMDHs from other organisms such as fish, plant, and mammal. But low sequence similarities have been found between this cMDH and mitochondrial malate dehydrogenase as well as glyoxysomal malate dehydrogenase from other organisms. Northern blot analysis showed the size of the C. sinensis cMDH mRNA was 1.2 kb. The cMDH was expressed in Escherichia coli M15 as a His-tag fusion protein and purified by BD TALON metal affinity column. The recombinant cMDH showed high MDH activity of 241 U mg(-1), without lactate dehydrogenase and NADP(H) selectivity. It provides a model for the structure, function analysis, and drug screening on cMDH. PMID- 15755420 TI - Schistosoma mansoni: functional proteasomes are required for development in the vertebrate host. AB - Proteasomes are multi-subunit proteases involved in several mechanisms and thought to contribute to the regulation of cellular homeostasis. Here, we report for the first time biochemical evidence for the existence of a ubiquitin proteasome proteolytic pathway in this parasite. Proteasomes from both cercariae and adult worms exhibited a high preference for hydrolysis of the substrate Suc LLVY-AMC, although in the cercariae extract the rate of hydrolysis was 50% lower when compared to adult worms extracts. The same difference in proteasome activities was observed when endogenous proteins were broken down in the presence of ATP and ubiquitin. Additionally, accumulation of high molecular weight conjugates was observed when cercariae were pre-incubated with proteasome inhibitors. Finally, we present evidence that during experimental schistosomiasis, proteasome inhibitors were able to reduce the number of lung stage schistosomula, reduce the worm burden and consequently decrease the egg output in infected mice. PMID- 15755421 TI - Echinococcus granulosus: lethal effect of low voltage direct electric current on hydatid cyst protoscoleces. AB - We have studied a small scale method for killing hydatid cyst protoscoleces using low voltage direct electric current. After collecting hydatid cysts from infected organs of slaughtered animals, protoscoleces were cultured in four different media: hydatid cyst fluid, RPMI, normal saline, and Tris buffer, respectively. Protoscoleces from each of the above media were then transferred to an electrolysis device through which different electric current densities were applied. For measuring the survival rate of protoscoleces, flame cell movement and eosin staining was used. The results show that the survival rate of protoscoleces in hydatid fluid was dependent on the electric current density and the time of the applied current. Current densities of 62.5 mA/cm2 (11 V), 53.71 mA/cm2 (10 V), and 18.18 mA/cm2 (5 V) after 1, 2, and 3 min, respectively, killed all the parasites in the hydatid fluid. However, a current density of 7 mA/cm2 (9 V) in RPMI medium after 3 min was most effective. PMID- 15755422 TI - Entamoeba histolytica: intracellular distribution of the sec61alpha subunit of the secretory pathway and down-regulation by antisense peptide nucleic acids. AB - The Sec61alpha protein is defined as a highly conserved essential integral component of the endoplasmic reticulum in eukaryotic cells. We report a detailed immunolocalization of the Entamoeba histolytica homologue of the Sec61alpha subunit (EhSec61alpha), which shows an irregular pattern throughout the cell and is also found on the cell surface, its effective down-regulation by means of antisense peptide nucleic acids and its effects on cell proliferation, subcellular distribution of two virulence factors, and the ability of the trophozoites to cause liver abscess in hamsters. Although Sec61alpha levels are specifically decreased in antisense PNA-treated trophozoites, which proliferate more slowly than the controls, mobilization of the cysteine protease 5 and amoebapore to the cell surface is not significantly impeded and the capacity to induce liver abscess in hamsters is largely unaffected. The implications of these findings are discussed in the context of the peculiar cell biology of E. histolytica. PMID- 15755423 TI - Setaria cervi: immunoprophylactic potential of glutathione-S-transferase against filarial parasite Brugia malayi. AB - Glutathione-S-transferase (GST) has been detected in the adult female Setaria cervi, a bovine filarial parasite. The role of S. cervi GST antigen in inducing immunity in the host against Brugia malayi microfilariae and infective larvae was studied by in vitro antibody dependent cell mediated reaction as well as in situ inoculation of filarial parasites within a microchamber in Mastomys. The immune sera from glutathione-S-transferase immunized Mastomys promoted the adherence of peritoneal exudate cells to B. malayi microfilariae and infective larvae in vitro inducing 80.7 and 77.6% cytotoxicity, respectively in 72 h. In the microchambers implanted in the immunized Mastomys host cells could migrate and adhere to the microfilariae and infective larvae and induced 77.8 and 75% cytotoxicity to B. malayi microfilariae and infective larvae in 72 h, respectively. These results suggest that native GST from S. cervi is effective in inducing protection against heterologous B. malayi filarial parasite and thus has potential in immunoprophylaxis. PMID- 15755424 TI - Plasmodium vivax: allele variants of the mdr1 gene do not associate with chloroquine resistance among isolates from Brazil, Papua, and monkey-adapted strains. AB - We describe here the sequence of the Plasmodium vivax mdr1 gene from 10 different isolates differing in chloroquine sensitivity. The deduced amino acid sequence of PvMDR1 shares more than 70% similarity with other malarial MDR proteins and it displays consensus motifs of an ABC family transporter including two transmembrane domains and two ATP binding cassettes. Similarity and dendrogram analyses revealed that sequences could be grouped according to their geographical origin. Within each geographical group however, no correlation was found between chloroquine resistance and specific mutations. PMID- 15755425 TI - Trypanosoma cruzi: molecular characterization of TcPUF6, a Pumilio protein. AB - In trypanosomes regulation of gene expression occurs mainly at the post transcriptional level. Pumilio proteins are RNA-binding proteins that modulate gene expression in lower and higher eukaryotes. Here we present the characterization of TcPUF6, a member of the Pumilio family in Trypanosoma cruzi. TcPUF6 is expressed in the different life cycle forms of the parasite showing no clear stage specific regulation and it is localized to multiple discrete foci in the cytoplasm of epimastigotes. The recombinant TcPUF6 fusion protein specifically binds to the Drosophila hunchback NRE (nanos response element). TcPUF6 conserves functional properties that characterize the Pumilio family throughout evolution. PMID- 15755426 TI - Outcome instruments for the assessment of the upper extremity following trauma: a review. AB - Orthopaedic injuries are common among trauma patients and can result in long-term problems. Considerable data are available regarding functional outcomes following lower extremity trauma. There is, however, a paucity of data available for upper extremity trauma patients. Whilst currently available instruments appear to assess outcomes of relevance in trauma populations, the reliability, validity and responsiveness of these instruments have not been evaluated in the upper extremity trauma population. This paper reviews instruments designed for patient self-evaluation of musculoskeletal disorders of the upper extremity, and instruments used in an orthopaedic trauma population to assess functional recovery following injury. The Musculoskeletal Functional Assessment (MFA), Short Musculoskeletal Functional Assessment (SMFA), Disabilities of the Arm, Shoulder, and Hand (DASH), American Shoulder and Elbow Surgeons Shoulder Scale (ASES-s), American Shoulder and Elbow Surgeons Elbow Scale (ASES-e), Patient Rated Elbow Evaluation (PREE), and the Patient Rated Wrist Evaluation (PRWE) were reviewed. Until research is published outlining the evaluation of assessment instruments in upper extremity orthopaedic populations, authors will need to conduct their own validation studies before investigating outcomes in specific trauma populations. PMID- 15755427 TI - The Injury Severity Score or the New Injury Severity Score for predicting intensive care unit admission and hospital length of stay? AB - OBJECTIVES: To compare the New Injury Severity Score (NISS) and the Injury Severity Score (ISS) as predictors of intensive care unit (ICU) admission and hospital length of stay (LOS) in an urban North American trauma population and in a subset of patients with head injuries. METHODS: The study population consisted of 23,909 patients from three urban level I trauma centres in the province of Quebec, Canada. The predictive accuracies of the NISS and the ISS were compared using Receiver Operator Characteristic (ROC) curves and Hosmer-Lemeshow (H-L) statistics for the logistic regression model of ICU admission and using r2 for the linear regression model of LOS. RESULTS: A total of 7660 (32%) patients were admitted to the ICU. Mean LOS was 8.2+/-2.5 days. In the whole sample, the NISS presented equivalent discrimination (area under ROC curve: NISS = 0.839 versus ISS = 0.843, p = 0.08) but better calibration (H-L statistic: 309 versus 611) for predicting ICU admission. In the subgroup patients with moderate to serious head injuries, the NISS was a better predictor of ICU admission in terms of both discrimination (area under ROC curve: NISS = 0.771 versus ISS = 0.747, p < 0.00001) and calibration (H-L statistic: 12 versus 21). The NISS explained more variation in LOS than the ISS for the whole sample (r2 = 0.254 versus 0.249, p = 0.0008) and in the sub-population with moderate to severe head injuries (r2 = 0.281 versus 0.263, p = 0.0002). CONCLUSIONS: The NISS is a better choice for case mix control in trauma research than the ISS for predicting ICU admission and LOS, particularly among patients with moderate to severe head injuries. PMID- 15755428 TI - Improvements in passive car safety led to decreased injury severity--a comparison between the 1970s and 1990s. AB - PURPOSE: The purpose of this study was to characterize changes in the mechanism and pattern of injury for vehicular trauma victims with modern vehicle design. Crash and injury severity were specifically investigated to isolate the influence of these improvements in vehicle design. METHODS: Since 1972, a local, prospective, assessment of vehicular trauma victims on-scene and at medical institutions providing care has been performed including the following parameters: delta-v, collision speed, type of road using, abbreviated injury scale (AIS), injury severity score (ISS), incidence of polytrauma or death. Victims (for restrained car occupants, bicyclists, pedestrians) injured between 1973 and 1978, and between 1994 and 1999 were compared. RESULTS: Lower crash severity (delta-v, collision speed) and injury severity (AIS, ISS, incidence of polytrauma or death) were measured for restrained car occupants, bicyclists and pedestrians during the later period. The correlation coefficient between delta-v or vehicle collision speed and ISS was higher in the earlier period for car occupants, cyclists and pedestrians. CONCLUSIONS: This study suggests that the observed reduction in injury severity in restrained car occupants, bicyclists and pedestrians is not only linked to the reduction of crash severity, but also related to improvements in vehicle design beyond seat-belt use. Passive car safety led to decreased injury severity--a comparison. PMID- 15755429 TI - Recombinant human bone morphogenetic protein 2 enhances bone healing in an experimental model of fractures at risk of non-union. AB - INTRODUCTION: Identification of patients at risk of developing non-union and institution of procedures preventing non-union could be attractive in routine fracture management. We investigated whether recombinant human bone morphogenetic protein (rhBMP-2) delivered in a hyaluronic acid carrier could prevent non-union development in an experimental non-union model, which simulates the clinical situation of open mid-tibial fractures. METHODS: Sixteen rabbits underwent a standard non-union operation comprising mid-tibial osteotomy, excision of periosteum and endosteum, and plate fixation. Before closure of the wound eight rabbits received interfragmentary deposition of 200 microg rhBMP-2 delivered in a hyaluronan gel carrier, and eight rabbits received gel carrier alone. RESULTS: After 7 weeks, torsional failure moment of the osteotomy and energy absorbed at failure, macroscopic and radiographic appearance, callus area, and interfragmentary bone volume fraction confirmed that rhBMP-2 delivery significantly improved bone healing. Blood flow at the osteotomy site, measured using radiolabelled microspheres, was not higher in the united osteotomies than in non-united osteotomies. DISCUSSION: rhBMP-2 delivered in a hyaluronic acid carrier-induced formation of competent bone in an experimental model of compromised healing. We, therefore, propose interfragmentary deposition of rhBMP 2 delivered in a hyaluronic acid carrier to patients encountering fractures at risk of non-union or delayed union. PMID- 15755431 TI - Independent lung ventilation for asymmetrical chest trauma: effect on ventilatory and haemodynamic parameters. AB - Complex thoracic injuries significantly alter the lung mechanics. There appears to be severe ventilation-perfusion inequality that is enhanced by the asymmetrical compliance of the injured areas. In cases of unilateral massive air leaks large tidal volumes are needed to deliver adequate air volume to the injured lung. In such cases, mechanical ventilation via a standard tracheal tube will direct a large fraction of the tidal volume to the less affected areas. An alternative means of ventilating these patients is the application of independent lung ventilation through a double lumen tracheal tube and the use of two separate ventilators. Two such cases are presented with emphasis given on the ventilatory and haemodynamic changes that were recorded. PMID- 15755432 TI - Pre-operative analgesia for patients with femoral neck fractures using a modified fascia iliaca block technique. AB - Adequate pre-operative analgesia for elderly patients with femoral neck fractures is difficult to assess and is often an overseen aspect of their care. We aimed to assess the efficacy of fascia iliaca blocks inserted via plexus blockade catheters in the pre-operative period. Our simple technique allowed the block to be administered safely without the need for a nerve stimulator. We assessed the effectiveness of the block with a novel objective sitting score and by assessing the degree of passive hip flexion that could be achieved comfortably. Visual analogue scores were also used. We studied 30 consecutive patients, regardless of their mental state. One hour following the block, there was a significant improvement in the sitting scores as well as the passive hip flexion (mean increase 44 degrees ). Visual analogue scores also score improved significantly from 7.2 to 4.6 (S.D. 2.4) in the 18 patients without cognitive impairment. We conclude that fascia iliaca blocks can provide significant benefit in the pre operative period and allow patients to sit up more comfortably while they await surgery. PMID- 15755433 TI - Coping strategies, injury characteristics and long-term outcome after burn injury. AB - Coping consists of cognitive and behavioural strategies aimed at enhancing adaptation, and the use of certain coping strategies is proposed to be beneficial for health. The relationship between coping strategies and burn injury characteristics, sociodemographics and long-term outcome was evaluated in 161 previous victims of severe burn injury. Functional and psychosocial restrictions were measured with the burn specific health scale-brief (BSHS-B) and related to coping strategies measured by the coping with burns questionnaire (CBQ). Patients were on average 47.7 years at measurement of health status and they were assessed 9.2 (S.D. = 4.8) years after injury. The mean area burned was 24.0 and 7.2% was full thickness injury. There was no relation between coping strategies and injury characteristics except in individuals with a full thickness burn exceeding 10%, who exhibited more Revaluation/adjustment. An Avoidant coping strategy was related to work status, marital status and living conditions, and this was the strategy most clearly related to "Bad outcome" in all scales of the BSHS-B. Emotional support was the most beneficial strategy and was mainly associated with the psychosocial scales of the BSHS-B. PMID- 15755430 TI - The role of recombinant factor VIIa in the treatment of life-threatening haemorrhage in blunt trauma. AB - BACKGROUND: Recombinant factor VIIa (rFVIIa) is a novel haemostatic agent originally developed to treat bleeding in haemophiliacs. Several case reports suggest effectiveness of rFVIIa in the treatment of patients without pre-existing bleeding disorders. The aim of this study is to evaluate treatment with recombinant (rFVIIa) in blunt trauma patients with uncontrolled bleeding. PATIENTS AND METHODS: This study was designed as a retrospective case review. Consecutive patients with life-threatening uncontrolled bleeding due to blunt trauma who were treated with rFVIIa were selected. Data were obtained from medical records. RESULTS: A total of eight blunt trauma patients were treated with rFVIIa for uncontrolled bleeding. After treatment the need for transfusion of red blood cells (RBC) decreased significantly from 31.3 +/- 15.8 to 6.1 +/- 6.8 units (P = 0.003), fresh frozen plasma (FFP) from 13.3 +/- 6.6 to 5 +/- 6.3 units (P = 0.02), and platelets from 3.6 +/- 1.8 to 1.5 +/- 2.3 units (P = 0.01). Three patients died of non-bleeding complications. The other five fully recovered. CONCLUSION: Treatment with rFVIIa reduced or stopped bleeding in all patients. No adverse events were registered. Prospective studies are mandatory to elucidate the role of rFVIIa in blunt trauma. PMID- 15755434 TI - Clinical evaluation of the spine in the intoxicated blunt trauma patient. AB - The objective of the current study was to evaluate the accuracy of the clinical examination of the spine following blunt trauma in intoxicated patients. Methods included a retrospective review of all cases of blunt trauma presenting to an urban level I trauma centre over 1 year. Data was analysed separately for the clinical examination of the cervical spine (CS) and for the thoracic and lumbar spine (T + LS). Two hundred and sixteen cases of blunt trauma secondary to MVC (n = 143) or falls (n = 73) were retained for analysis. In intoxicated patients, sensitivities and specificities for CS tenderness were 60.0 and 64.3% (radiological abnormality) and 100 and 68.6% (operative stabilization), respectively. With respect to the T + LS in intoxicated patients; sensitivities and specificities for T + LS tenderness and radiological abnormality were 80.0 and 71.4% and for the ability of the clinical examination to pick up unstable T + LS fractures requiring operative stabilization 100 and 72.0%, respectively. Intoxicated blunt trauma patients may be able to have spine fractures requiring operative stabilization excluded using physical examination of the spine at presentation to the trauma center. PMID- 15755435 TI - Videothoracoscopy versus intrapleural streptokinase for management of post traumatic retained haemothorax: a retrospective study of 65 cases. AB - BACKGROUND: Post traumatic retained haemothorax (PRH) may cause pulmonary restrictions or septic pleural complications. Currently, minimally invasive procedures such as videothoracoscopy or intrapleural fibrinolysis have replaced open surgery in an effort to avoid these complications. OBJECTIVE: We have reviewed retrospectively our use of videothoracoscopy versus intrapleural streptokinase for the management of PRH over the last 10 years. PATIENTS AND METHOD: There were 56 males and nine females in the study. Thirty-one cases had been managed by intrapleural streptokinase (group I), and videothoracoscopy was performed on 34 cases (group II). Therapeutic results for both groups were determined by chest radiographs. RESULTS: In the population from which we drew our study group, retained haemothoraces occurred in 10.9% of 596 cases with traumatic haemothorax. In group I, 22 patients had radiological improvement; the others underwent thoracotomy. In this group, mean hospitalisation time was 14.5(+/-4.2) days, and three cases were complicated by empyema. In group II, all patients except four had complete radiological improvement; two of them required a decortication. In this group, mean hospital stay was 9.8(+/-3.7) days. There were no deaths in either group. The differences between group I and group II for length of hospital stay and number of thoracotomies was statistically significant. CONCLUSION: Videothoracoscopy is therefore a more effective procedure than intrapleural streptokinase for the management of PRH. PMID- 15755436 TI - Minimally invasive plate osteosynthesis (MIPO) of the humeral shaft fracture. Is it possible? A cadaveric study and preliminary report. AB - Minimally Invasive Plate Osteosynthesis (MIPO) has gained popularity with satisfactory clinical outcomes in the treatment of long bone fractures. MIPO for humeral shaft fractures, however, could be a surgically dangerous procedure because of the risk of radial nerve injury. An anatomical study was performed to evaluate the feasibility of MIPO for the humeral shaft fractures, and to study the relationship between the radial nerve and the plate with the forearm in full pronation and in supination. The study was performed on ten arms from five fresh cadavers. Two separate incisions, one proximal and one distal, were made in each arm with the forearm in full supination. A 9-hole narrow DCP was inserted into a tunnel using an anterior approach and fixed with 2 screws each on the proximal and distal humerus. The tunnel was then explored to identify the relationship between the radial nerve and the plate. No radial nerve compression or entrapment by the plate was found. The distance measured from the closest part of the plate to the radial nerve was 2.0-4.9 mm (average 3.2 mm). When the forearm was pronated, the radial nerve moved closer to the plate by a distance of 0-3 mm. The results of this study showed that it is possible to treat humeral shaft fractures by the MIPO method using an anterior approach. To reduce the risk of radial nerve injury, the forearm must be kept in full supination during plate insertion, and excessive force should be avoided during retraction of the lateral half of the brachialis muscle together with the radial nerve in the distal incision. The results of using this MIPO approach for humeral shaft fractures in 4 patients were also reviewed. PMID- 15755437 TI - Outcome following Bankart repair for shoulder instability 1998-2002 using a subjective patient based shoulder questionnaire (Oxford). AB - Bankart repair for shoulder instability has been performed by the senior author since 1998. We employ a modification of the technique as described by Rowe et al., using suture anchors but without a coracoid osteotomy. If inferior instability was present this procedure was combined with a capsular shift. We performed a retrospective case note review and postal questionnaire incorporating the Oxford Instability Score (OIS) on all patients. Fifty Bankart repairs were performed in the study period, three patients had further dislocations and three patients had ongoing symptoms of instability. Response rate to the questionnaire was 62%. Mean OIS following primary stabilisation was 21.7 (possible scores from 12-excellent to 60-poor). OIS following Bankart stabilisation of the shoulder has not been previously reported. These results compare favourably to original scores published by Dawson et al. (1999) who included both patients treated by surgery or physiotherapy. PMID- 15755438 TI - Secondary effects of femoral instrumentation on pulmonary physiology in a standardised sheep model: what is the effect of lung contusion and reaming? AB - Intramedullary nailing is the treatment of choice for patients with femoral shaft fractures. However, there is an ongoing debate in multiple trauma patients with associated lung contusion when primary or secondary definitive stabilisation of the long bone fracture should be performed, as nailing is thought to play an important role in the pathogenesis of adult respiratory distress syndrome (ARDS). In a standardised sheep model, this study aimed to quantify the development of acute pulmonary endothelial changes, to assess the activation of polymorphonuclear leucocytes (PMNL) and to observe the effects on the coagulation system associated with the reamed nailing procedure. Furthermore, the effect of coexisting lung contusion in an experimental model was evaluated. The animals were randomly assigned to one of four different groups (6 animals/group). In control groups, only a sham operation (thoracotomy) was performed, whereas in study groups, lung contusion was induced prior to femoral stabilisation either by external fixation or reamed femoral nailing. Using bronchoalveolar lavage (BAL) pulmonary permeability changes were quantified and PMNL activation was assessed by chemiluminescence. Additionally PMNL diapedesis and interstitial lung oedema were determined by histological analysis. All animals were sacrificed 4 h after the start of the femoral instrumentation. Without an associated lung injury, instrumentation of the femoral canal with the reamed nailing technique induced a transient increase in pulmonary permeability. In the face of an induced lung contusion, reamed femoral nailing resulted in significant increases in PMNL activation, pulmonary permeability and interstitial lung oedema, compared with external fixation. Without pulmonary contusion, reaming of the femoral canal was associated with a transient increase in pulmonary permeability. This was exacerbated in the presence of lung contusion along with increased PMNL activation. External fixation did not provoke similar changes. The findings of this study support the view that reaming of the femoral canal should be avoided in polytrauma patients with severe chest trauma as it could act as an additional stimulus for adverse outcome. Temporary external fixation appears to be a safe method for fracture stabilisation until inflammatory and coagulatory disturbances after trauma have been normalized. PMID- 15755440 TI - The use of multi-media messaging in the referral of musculoskeletal limb injuries to a tertiary trauma unit using: a 1-month evaluation. AB - OBJECTIVES: New developments in telecommunications will have a profound effect on the delivery of medical care throughout the world. In Northern Ireland three trauma centres provide fracture care for their own and surrounding Emergency Departments. All trauma referrals are currently taken by telephone. It is our experience that the verbal description of the radiographs of a musculoskeletal limb injury can be inaccurate, necessitating us to view the plain films of the patient. By utilising a recent advance in telecommunications technology, the launch of mobile handsets with multi-media messaging (MMS) service capability, it is now possible to digitally capture and instantly send an image of a plain film. PURPOSE: To evaluate the use of multi-media messaging as a supplement to the telephone referral of musculoskeletal limb injures. METHOD: : Following a referral using, the emergency physician and the trauma surgeon evaluated the multi-media consult through a survey questionnaire. RESULTS: Between the 1st December 2003 and the 1st January 2004, 46 multi-media consultations were performed. Picture quality was acceptable in all but one of the referrals. In 35 of the 46 referrals the multi-media image of the plain films was felt to improve the management of the patient. In 8 of the 46 referrals the multi-media image of the plain films was felt to change the management of the patient. CONCLUSION: A multi-media messaging store- and forward telemedicine system has potential to facilitate the rapid, cost-effective management of musculoskeletal limb injuries thereby enhancing clinical care. PMID- 15755439 TI - Reproducibilty of partial weight bearing. AB - OBJECTIVES: To find out whether partial weight bearing can be reproduced and retained. DESIGN: In vivo experiment in normal subjects. INTERVENTION: Training for partial weight bearing (25% of body weight) using bathroom scales. MAIN OUTCOME MEASUREMENT: Reproducibility on force platform immediately after training and after 60 min. RESULTS: Twelve subjects were asked to reproduce 25% of their body weight through either the dominant or non-dominant limb on force platform after three practice attempts on bathroom scales with concurrent visual feedback. No feedback was provided after the measurements on force plate. The process was repeated after 1h without any practice sessions in the interim period to find out if the weight practised could be retained. The mean 0-min reading was found to be 25.9% of body weight while the mean 60-min reading was found to be 24.4%. The p value for the difference between the two means was found to be 0.3841. CONCLUSIONS: This study indicates that partial weight bearing instructions can be quantified and graded. Simple bathroom scales are sufficient to educate the patients and this can be practised at home after an initial period of supervision. PMID- 15755441 TI - Avulsion fracture of the coracoid process: a case report. PMID- 15755442 TI - Falling downstairs does not mean a fracture: the 1st case report of an immunocompetent community acquired MRSA disc/psoas abscess. PMID- 15755443 TI - Bridge plate osteosynthesis of humeral shaft fractures. PMID- 15755444 TI - Correlation between omega and psi dihedral angles in protein structures. AB - The planarity of the peptide group is one of the fundamental features of protein structure that is described in every chemistry and biochemistry textbook. By surveying a dataset of 163 atomic resolution protein structures we here identify the stereochemical conditions that favor significant deformations of peptide bond planarity. In particular, we demonstrate that the values of the omega dihedral angle are strictly correlated to the values of the adjacent psi angle. This trend is also observed in highly strained states such as those occurring in vicinal disulfide bridges. These findings provide direct evidence for the mutual influence of the geometrical parameters that describe the protein structure. PMID- 15755445 TI - Intramolecular cooperativity in a protein binding site assessed by combinatorial shotgun scanning mutagenesis. AB - Combinatorial shotgun alanine-scanning was used to assess intramolecular cooperativity in the high affinity site (site 1) of human growth hormone (hGH) for binding to its receptor. A total of 19 side-chains were analyzed and statistically significant data were obtained for 145 of the 171 side-chain pairs. The analysis revealed that 90% of the side-chain pairs exhibited no statistically significant pair interactions, and the remaining 10% of side-chain pairs exhibited only small interactions corresponding to cooperative interaction energies with magnitudes less than 0.4 kcal/mol. The statistical predictions were tested by measuring affinities for purified mutant proteins and were found to be accurate for five of six side-chain pairs tested. The results reveal that hGH site 1 behaves in a highly additive manner and suggest that shotgun scanning should be useful for assessing cooperative effects in other protein-protein interactions. PMID- 15755446 TI - Differential DNA binding and protection by dimeric and dodecameric forms of the ferritin homolog Dps from Deinococcus radiodurans. AB - Bacterial iron storage proteins such as ferritin serve as intracellular iron reserves. Members of the DNA protection during starvation (Dps) family of proteins are structurally related to ferritins, and their function is to protect the genome from iron-induced free radical damage. Some members of the Dps family bind DNA and are thought to do so only as fully assembled dodecamers. We present the cloning and characterization of a Dps homolog encoded by the radiation resistant eubacterium Deinococcus radiodurans and show that DNA binding does not require its assembly into a dodecamer. D.radiodurans Dps-1, the product of gene DR2263, adopts a stably folded conformation, as demonstrated by circular dichroism spectroscopy, and undergoes a transition to a disordered state with a melting temperature of 69.2(+/-0.1) degrees C. While a dimeric form of Dps-1 is observed under low-salt conditions, a dodecameric assembly is highly favored at higher concentrations of salt. Both oligomeric forms of Dps-1 exhibit ferroxidase activity, and Fe(II) oxidation/mineralization is seen for dodecameric Dps-1. Notably, addition of Ca(2+) (to millimolar concentrations) to dodecameric Dps-1 can result in the reduction of bound Fe(III). Dimeric Dps-1 protects DNA from both hydroxyl radical cleavage and from DNase I-mediated cleavage; however, dodecameric Dps-1 is unable to provide efficient protection against hydroxyl radical-mediated DNA cleavage. While dodecameric Dps-1 does bind DNA, resulting in formation of large aggregates, cooperative DNA binding by dimeric Dps-1 leads to formation of protein-DNA complexes of finite stoichiometry. PMID- 15755447 TI - Uncoupling of unwinding from DNA synthesis implies regulation of MCM helicase by Tof1/Mrc1/Csm3 checkpoint complex. AB - The replicative DNA helicases can unwind DNA in the absence of polymerase activity in vitro. In contrast, replicative unwinding is coupled with DNA synthesis in vivo. The temperature-sensitive yeast polymerase alpha/primase mutants cdc17-1, pri2-1 and pri1-m4, which fail to execute the early step of DNA replication, have been used to investigate the interaction between replicative unwinding and DNA synthesis in vivo. We report that some of the plasmid molecules in these mutant strains became extensively negatively supercoiled when DNA synthesis is prevented. In contrast, additional negative supercoiling was not detected during formation of DNA initiation complex or hydroxyurea replication fork arrest. Together, these results indicate that the extensive negative supercoiling of DNA is a result of replicative unwinding, which is not followed by DNA synthesis. The limited number of unwound plasmid molecules and synthetic lethality of polymerase alpha or primase with checkpoint mutants suggest a checkpoint regulation of the replicative unwinding. In concordance with this suggestion, we found that the Tof1/Csm3/Mrc1 checkpoint complex interacts directly with the MCM helicase during both replication fork progression and when the replication fork is stalled. PMID- 15755448 TI - Self-association properties of the bacteriophage lambda terminase holoenzyme: implications for the DNA packaging motor. AB - Terminases are enzymes common to complex double-stranded DNA viruses and are required for packaging of viral DNA into a protective capsid. Bacteriophage lambda terminase holoenzyme is a hetero-oligomer composed of the A and Nu1 lambda gene products; however, the self-association properties of the holoenzyme have not been investigated systematically. Here, we report the results of sedimentation velocity, sedimentation equilibrium, and gel-filtration experiments studying the self-association properties of the holoenzyme. We find that purified, recombinant lambda terminase forms a homogeneous, heterotrimeric structure, consisting of one gpA molecule associated with two gpNu1 molecules (114.2 kDa). We further show that lambda terminase adopts a heterogeneous mixture of higher-order structures, with an average molecular mass of 528(+/-34) kDa. Both the heterotrimer and the higher-order species possess site-specific cos cleavage activity, as well as DNA packaging activity; however, the heterotrimer is dependent upon Escherichia coli integration host factor (IHF) for these activities. Furthermore, the ATPase activity of the higher-order species is approximately 1000-fold greater than that of the heterotrimer. These data suggest that IHF bending of the duplex at the cos site in viral DNA promotes the assembly of the heterotrimer into a biologically active, higher-order packaging motor. We propose that a single, higher-order hetero-oligomer of gpA and gpNu1 functions throughout lambda development. PMID- 15755449 TI - Unique iron binding and oxidation properties of human mitochondrial ferritin: a comparative analysis with Human H-chain ferritin. AB - Ferritins are ubiquitous iron mineralizing and storage proteins that play an important role in iron homeostasis. Although excess iron is stored in the cytoplasm, most of the metabolically active iron is processed in the mitochondria of the cell. Little is known about how these organelles regulate iron homeostasis and toxicity. The recently discovered human mitochondrial ferritin (MtF), unlike other mammalian ferritins, is a homopolymer of 24 subunits that has a high degree of sequence homology with human H-chain ferritin (HuHF). Parallel experiments with MtF and HuHF reported here reveal striking differences in their iron oxidation and hydrolysis chemistry despite their similar diFe ferroxidase centers. In contrast to HuHF, MtF does not regenerate its ferroxidase activity after oxidation of its initial complement of Fe(II) and generally has considerably slower ferroxidation and mineralization activities as well. MtF exhibits sigmoidal kinetics of mineralization more characteristic of an L-chain than an H-chain ferritin. Site-directed mutagenesis reveals that serine 144, a residue situated near the ferroxidase center in MtF but absent from HuHF, is one player in this impairment of activity. Additionally only one-half of the 24 ferroxidase centers of MtF are functional, further contributing to its lower activity. Stopped-flow absorption spectrometry of Fe(II) oxidation by O(2) in MtF shows the formation of a transient diiron(III) mu-peroxo species (lambda(max) = 650 nm) as observed in HuHF. Also, as for HuHF, minimal hydroxyl radical is produced during the oxidative deposition of iron in MtF using O(2) as the oxidant. However, the 2Fe(II) + H(2)O(2) detoxification reaction found in HuHF does not occur in MtF. The structural differences and the physiological implications of the unique iron oxidation properties of MtF are discussed in light of these results. PMID- 15755450 TI - Intriguing conformation changes associated with the trans/cis isomerization of a prolyl residue in the active site of the DsbA C33A mutant. AB - Escherichia coli DsbA belongs to the thioredoxin family and catalyzes the formation of disulfide bonds during the folding of proteins in the bacterial periplasm. It active site (C30-P31-H32-C33) consists of a disulfide bridge that is transferred to newly translocated proteins. The work reported here refers to the DsbA mutant termed C33A that retains, towards reduced unfolded thrombin inhibitor, an activity comparable with the wild-type enzyme. Besides, C33A is also able to form a stable covalent complex with DsbB, the membrane protein responsible for maintaining DsbA in its active form. We have determined the crystal structure of C33A at 2.0 angstroms resolution. Although the general architecture of wt DsbA is conserved, we observe the trans/cis isomerization of P31 in the active site and further conformational changes in the so-called "peptide binding groove" region. Interestingly, these modifications involve residues that are specific to DsbA but not to the thioredoxin family fold. The C33A crystal structure exhibits as well a hydrophobic ligand bound close to the active site of the enzyme. The structural analysis of C33A may actually explain the peculiar behavior of this mutant in regards with its interaction with DsbB and thus provides new insights for understanding the catalytic cycle of DsbA. PMID- 15755451 TI - Using a library of structural templates to recognise catalytic sites and explore their evolution in homologous families. AB - Catalytic site structure is normally highly conserved between distantly related enzymes. As a consequence, templates representing catalytic sites have the potential to succeed at function prediction in cases where methods based on sequence or overall structure fail. There are many methods for searching protein structures for matches to structural templates, but few validated template libraries to use with these methods. We present a library of structural templates representing catalytic sites, based on information from the scientific literature. Furthermore, we analyse homologous template families to discover the diversity within families and the utility of templates for active site recognition. Templates representing the catalytic sites of homologous proteins mostly differ by less than 1A root mean square deviation, even when the sequence similarity between the two proteins is low. Within these sets of homologues there is usually no discernible relationship between catalytic site structure similarity and sequence similarity. Because of this structural conservation of catalytic sites, the templates can discriminate between matches to related proteins and random matches with over 85% sensitivity and predictive accuracy. Templates based on protein backbone positions are more discriminating than those based on side-chain atoms. These analyses show encouraging prospects for prediction of functional sites in structural genomics structures of unknown function, and will be of use in analyses of convergent evolution and exploring relationships between active site geometry and chemistry. The template library can be queried via a web server at and is available for download. PMID- 15755452 TI - Structure of MFP2 and its function in enhancing MSP polymerization in Ascaris sperm amoeboid motility. AB - The simplicity and specialization of the cell motility machinery of Ascaris sperm provides a powerful system in which to probe the basic molecular mechanism of amoeboid cell motility. Although Ascaris sperm locomotion closely resembles that seen in many other types of crawling cell, movement is generated by modulation of a cytoskeleton based on the major sperm protein (MSP) rather than the actin present in other cell types. The Ascaris motility machinery can be studied conveniently in a cell-free in vitro system based on the movement of plasma membrane vesicles by fibres constructed from bundles of MSP filaments. In addition to ATP, MSP and a plasma membrane protein, reconstitution of MSP motility in this cell-free extract requires cytosolic proteins to orchestrate the site-specific assembly and bundling of MSP filaments that generates locomotion. One of these proteins, MFP2, accelerates the rate of movement in this assay. Here, we describe crystal structures of two isoforms of MFP2 and show that both are constructed from two domains that have the same fold based on a novel, compact beta sheet arrangement. Patterns of conservation observed in a structure based analysis of MFP2 sequences from different nematode species identified regions that may be putative functional interfaces involved both in interactions between MFP2 domains and also with other components of the sperm motility machinery. Analysis of the growth of fibres in vitro in the presence of added MFP2 indicated that MFP2 increases the rate of locomotion by enhancing the effective rate of MSP filament polymerization. This observation, together with the structural data, suggests that MFP2 may function in a manner analogous to formins in actin-based motility. PMID- 15755453 TI - Single molecule studies of antibody-antigen interaction strength versus intra molecular antigen stability. AB - We investigated molecular recognition of antibodies to membrane-antigens and extraction of the antigens out of membranes at the single molecule level. Using dynamic force microscopy imaging and enzyme immunoassay, binding of anti-sendai antibodies to sendai-epitopes genetically fused into bacteriorhodopsin molecules from purple membranes were detected under physiological conditions. The antibody/antigen interaction strength of 70-170 pN at loading rates of 2-50 nN/second yielded a barrier width of x = 0.12 nm and a kinetic off-rate (corresponding to the barrier height) of k(off) = 6s(-1), respectively. Bacteriorhodopsin unfolding revealed a characteristic intra-molecular force pattern, in which wild-type and sendai-bacteriorhodopsin molecules were clearly distinguishable in their length distributions, originating from the additional 13 amino acid residues epitope in sendai purple membranes. The inter-molecular antibody/antigen unbinding force was significantly lower than the force required to mechanically extract the binding epitope-containing helix pair out of the membrane and unfold it (126 pN compared to 204 pN at the same loading rate), meeting the expectation that inter-molecular unbinding forces are weaker than intra-molecular unfolding forces responsible for stabilizing native conformations of proteins. PMID- 15755454 TI - Redox-dependent structural reorganization in putidaredoxin, a vertebrate-type [2Fe-2S] ferredoxin from Pseudomonas putida. AB - Putidaredoxin (Pdx), a vertebrate-type [2Fe-2S] ferredoxin from Pseudomonas putida, transfers electrons from NADH-putidaredoxin reductase to cytochrome P450cam. Pdx exhibits redox-dependent binding affinities for P450cam and is thought to play an effector role in the monooxygenase reaction catalyzed by this hemoprotein. To understand how the reduced form of Pdx is stabilized and how reduction of the [2Fe-2S] cluster affects molecular properties of the iron-sulfur protein, crystal structures of reduced C73S and C73S/C85S Pdx were solved to 1.45 angstroms and 1.84 angstroms resolution, respectively, and compared to the corresponding 2.0 angstroms and 2.03 angstroms X-ray models of the oxidized mutants. To prevent photoreduction, the latter models were determined using in house radiation source and the X-ray dose received by Pdx crystals was significantly decreased. Structural analysis showed that in reduced Pdx the Cys45 Ala46 peptide bond flip initiates readjustment of hydrogen bonding interactions between the [2Fe-2S] cluster, the Sgamma atoms of the cysteinyl ligands, and the backbone amide nitrogen atoms that results in tightening of the Cys39-Cys48 metal cluster binding loop around the prosthetic group and shifting of the metal center toward the Cys45-Thr47 peptide. From the metal center binding loop, the redox changes are transmitted to the linked Ile32-Asp38 peptide triggering structural rearrangement between the Tyr33-Asp34, Ser7-Asp9 and Pro102-Asp103 fragments of Pdx. The newly established hydrogen bonding interactions between Ser7, Asp9, Tyr33, Asp34, and Pro102, in turn, not only stabilize the tightened conformation of the [2Fe-2S] cluster binding loop but also assist in formation of a specific structural patch on the surface of Pdx that can be recognized by P450cam. This redox-linked change in surface properties is likely to be responsible for different binding affinity of oxidized and reduced Pdx to the hemoprotein. PMID- 15755455 TI - The crystal structure of human PAPS synthetase 1 reveals asymmetry in substrate binding. AB - The high energy sulfate donor 3'-phosphoadenosine-5-phosphosulfate (PAPS) is used for sulfate conjugation of extracellular matrix, hormones and drugs. Human PAPS synthetase 1 catalyzes two subsequent reactions starting from ATP and sulfate. First the ATP sulfurylase domain forms APS, then the APS kinase domain phosphorylates the APS intermediate to PAPS. Up to now the interaction between the two enzymatic activities remained elusive, mainly because of missing structural information. Here we present the crystal structure of human PAPSS1 at 1.8 angstroms resolution. The structure reveals a homodimeric, asymmetric complex with the shape of a chair. The two kinase domains adopt different conformational states, with only one being able to bind its two substrates. The asymmetric binding of ADP to the APS kinase is not only observed in the crystal structure, but can also be detected in solution, using an enzymatic assay. These observations strongly indicate structural changes during the reaction cycle. Furthermore crystals soaked with ADP and APS could be prepared and the corresponding structures could be solved. PMID- 15755456 TI - Molecular recognition of human eosinophil-derived neurotoxin (RNase 2) by placental ribonuclease inhibitor. AB - Placental ribonuclease inhibitor (RI) binds diverse mammalian RNases with dissociation constants that are in the femtomolar range. Previous studies on the complexes of RI with RNase A and angiogenin revealed that RI utilises largely distinctive interactions to achieve high affinity for these two ligands. Here we report a 2.0 angstroms resolution crystal structure of RI in complex with a third ligand, eosinophil-derived neurotoxin (EDN), and a mutational analysis based on this structure. The RI-EDN interface is more extensive than those of the other two complexes and contains a considerably larger set of interactions. Few of the contacts present in the RI-angiogenin complex are replicated; the correspondence to the RI-RNase A complex is somewhat greater, but still modest. The energetic contributions of various interface regions differ strikingly from those in the earlier complexes. These findings provide insight into the structural basis for the unusual combination of high avidity and relaxed stringency that RI displays. PMID- 15755457 TI - Determination of ultrafast protein folding rates from loop formation dynamics. AB - Quenching of the triplet state of tryptophan by contact with cysteine can be used to measure the kinetics of loop formation in unfolded proteins. Here we show that cysteine quenching dynamics also provide a novel method for measuring folding rates when the exchange between folded and unfolded states is faster than the unquenched triplet lifetime (approximately 100 micros). We use this technique to investigate folding/unfolding kinetics of the 35 residue headpiece subdomain of the protein villin, which contains a single tryptophan residue and was engineered to contain a cysteine residue at the N terminus. At intermediate concentrations of denaturant the time-course of the triplet decay consists of two relaxations, the rates and amplitudes of which reveal the fast kinetics for folding and unfolding of this protein. The folding rates extracted using a simple kinetic model are close to those reported previously from laser-induced temperature-jump experiments that employ the change in tryptophan fluorescence as a probe. However, the results differ significantly from those reported from dynamic NMR line shape analysis on a variant with methionine at the N terminus, an issue that remains to be resolved. The analysis of the triplet quenching kinetics also shows that the quenching rates in the unfolded state increase with decreasing denaturant concentration, indicating a compaction of the unfolded protein. PMID- 15755458 TI - Sequential generation of two structurally distinct ovine prion protein soluble oligomers displaying different biochemical reactivities. AB - In pathologies due to protein misassembly, low oligomeric states of the misfolded proteins rather than large aggregates play an important biological role. In prion diseases the lethal evolution is associated with formation of PrP(Sc), a misfolded and amyloid form of the normal cellular prion protein PrP. Although several molecular mechanisms were proposed to account for the propagation of the infectious agent, the events responsible for cell death are still unclear. The correlation between PrP(C) expression level and the rate of disease evolution on one side, and the fact that PrP(Sc) deposition in brain did not strictly correlate with the apparition of clinical symptoms on the other side, suggested a potential role for diffusible oligomers in neuronal death. To get better insight into the molecular mechanisms of PrP(C) oligomerization, we studied the heat induced oligomerization pathway of the full-length recombinant ovine PrP at acidic pH. This led to the irreversible formation of two well-identified soluble oligomers that could be recovered by size-exclusion chromatography. Both oligomers displayed higher beta-sheet content when compared to the monomer. A sequential two-step multimolecular process accounted for the rate of their formation and their ratio partition, both depending on the initial protein concentration. Small-angle X-ray scattering allowed the determination of the molecular masses for each oligomer, 12mer and 36mer, as well as their distinct oblate shapes. The two species differed in accessibility of polypeptide chain epitopes and of pepsin-sensitive bonds, in a way suggesting distinct conformations for their monomeric unit. The conversion pathway leading to these novel oligomers, displaying contrasted biochemical reactivities, might be a clue to unravel their biological roles. PMID- 15755460 TI - Epidemiology and risk factors for chronic kidney disease. AB - Kidney disease is highly prevalent in the United States population and groups at high risk for increased prevalence of CKD include individuals with a family history of ESRD, diabetes, hypertension, and cardiovascular disease. Despite the increased risk of ESRD observed for blacks compared with whites, racial disparities in the prevalence of kidney disease have not been consistently demonstrated in the United States population. Although the reasons for discrepancy in risk of ESRD and CKD have not been established, clinicians should be aware that more rapid progression of CKD among blacks is a possible explanation for this observation and that closer monitoring and intensive care of risk factors associated with progressive renal injury is warranted for blacks with CKD and in other high-risk groups. Therapeutic interventions that delay or prevent progressive kidney disease are well established and incorporated into widely disseminated clinical practice guidelines. These interventions include aggressive blood pressure control with agents that block the renin-angiotensin system, reduction of dietary protein to recommended levels for the American diet, weight loss, smoking cessation, and control of hyperlipidemia. These interventions also reduce the risk of cardiovascular disease and should be regarded as essential components of care of CKD. Achieving high levels of medically appropriate care of CKD patients and reduction in risk of progression to ESRD may be delayed by barriers created by individual and regional poverty. PMID- 15755461 TI - The familial clustering of renal disease and related phenotypes. AB - This article reviews the familial aggregation of chronic kidney diseases including end-stage renal disease and albuminuria, along with variation in glomerular filtration rate. In addition to environmental influences on the progression of nephropathy, epidemiologic evidence in support of the existence of renal failure susceptibility genes is presented. PMID- 15755462 TI - Measurement of kidney function. AB - Accurate estimation of kidney function is central to the detection, evaluation, and treatment of chronic kidney disease (CKD). Glomerular filtration rate (GFR) is widely accepted as the best overall measure of kidney function and as such it forms the basis of definition and classification system for CKD. GFR cannot be measured directly in clinical practice and multiple methods are used to estimate GFR, all of which have their strengths and limitations. The current recommended method is to use estimating equations based on serum creatinine and which incorporate demographic and clinical variables. PMID- 15755463 TI - Racial disparities in the optimal delivery of chronic kidney disease care. AB - This article provides evidence that the current and growing burden of CKD in racial and ethnic minority populations is likely to be multifactorial involving the interplay of biologic, clinical, social, and behavioral determinants. To eliminate these disparities, crafting successful solutions requires more attention to the constellation of contributing factors not only by specialists, primary care physicians, and other health care providers involved in CKD care, but also clinical and behavioral scientists, payers of health care, and patients. PMID- 15755464 TI - Strategies to retard the progression of chronic kidney disease. AB - Chronic kidney disease (CKD) is a common and, in advanced cases, highly morbid disorder. The most severe form of CKD is end-stage renal disease (ESRD), in which the patient requires some form of renal replacement therapy to survive. The increasing incidence, prevalence, and costs of ESRD are major national health care concerns; interventions that may prevent or slow the progression of CKD towards ESRD are extremely important and the focus of this article. PMID- 15755465 TI - Multiple risk factor intervention in chronic kidney disease: management of cardiac disease in chronic kidney disease patients. AB - This article describes the relationship between CVD and CKD, the current state of knowledge regarding medical interventions, and underscores the importance of attending to both CVD and kidney disease aspects in each individual. The burden of cardiac disease in CKD patients is high with severe LVH, dilated cardiomyopathy and coronary artery disease occurring frequently. This predisposes to congestive heart failure, angina, myocardial infarction, and death. Multiple risk factors for cardiac disease exist and include hypertension, diabetes, smoking, anemia, abnormal calcium and phosphate metabolism, inflammation, and LVH. The efficacy of risk factor intervention has not been established in these populations, although there is good evidence for good blood pressure control, partial correction of anemia, treatment of dyslipidemia, cessation of tobacco use, correction of divalent abnormalities, and aspirin us. Appropriate use of ACE inhibitors, beta-blockers, and statins should be encouraged. PMID- 15755466 TI - Etiology and management of hypertension in chronic kidney disease. AB - The kidneys are vital in the pathogenesis of hypertension and are also pathologically affected by the presence of hypertension. The prevalence of hypertension in chronic kidney disease (CKD) depends on age, the severity of renal failure, and proteinuria. The intricate and inextricable relationship between CKD and hypertension seems to cause cardiovascular disease that has assumed epidemic proportions. This article discusses the etiology and treatment of hypertension in CKD so that it can be better controlled. PMID- 15755467 TI - Complications of chronic kidney disease: anemia, mineral metabolism, and cardiovascular disease. AB - This article focuses on the importance of three major complications of chronic kidney disease: (1) anemia, (2) calcium-phosphorus regulation and bone disease, and (3) cardiovascular risk profiling and treatment. The arguments for early and effective intervention have been amply made with respect to these three complications. Substantive trial data are sorely need to provide the definitive evidence that effective treatment of these complications results in better outcomes. PMID- 15755468 TI - Treatment of acute coronary syndromes in patients who have chronic kidney disease. AB - Patients with CKD and CAD have traditionally been a difficult population to diagnose and treat in the setting of ACS. In addition to having poorer outcomes post-ACS, data are lacking regarding best treatments available. Aggressive interventional and medical treatments in this group with already poor outcomes are not necessarily contraindicated and should always be considered. The appalling outcome for CKD patients post-ACS is improved by many therapies shown to benefit in the non-CKD patients. Data suggest that troponins are useful markers in CKD patients, that major bleeding is not increased with the use of GP IIb-IIIa antagonists, that thrombolytics have been used successfully in CKD patients, and that PCI electively and as a primary treatment for ACS is successful and probably more beneficial to treatment. PMID- 15755469 TI - Traditional and nontraditional cardiovascular risk factors in chronic kidney disease. AB - Chronic kidney disease (CKD) is public health problem, with as many as 20 million individuals affected in the United States. Patients with CKD should be considered in the highest-risk group for development of cardiovascular disease (CVD), and aggressive treatment of traditional and nontraditional risk factors should be instituted. Additional randomized controlled trials are urgently needed to evaluate potential treatments in this population. This article focuses attention on the major modifiable cardiovascular risk factors in CKD. PMID- 15755470 TI - Gender differences in hypertension and kidney disease. AB - Gender differences exist in hypertension, the prevalence of several renal diseases, progression of established renal disease, and within the ESRD program, including renal transplantation. Sex hormones play key roles in the pathogenesis and outcome of disease processes. Observational data suggest gender differences in the prevalence and outcome of several renal diseases. The molecular mechanisms associated with physiologic phenomena needed to explain gender differences in renal disorders, however, remain largely obscure. The interaction of psychosocial, economic, medical, and genetic differences associated with discrepancies between the genders in the process of receiving a renal transplant and sustaining graft function are currently unclear. Additional studies are needed in these and other areas to explain gender differences in the incidence, prevalence, and outcome of renal disease. PMID- 15755472 TI - Drug dosing in chronic kidney disease. AB - Patients with chronic kidney disease (CKD) are at high risk for adverse drug reactions and drug-drug interactions. Drug dosing in these patients often proves to be a difficult task. Renal dysfunction-induced changes in human pathophysiology regularly results may alter medication pharmacodynamics and handling. Several pharmacokinetic parameters are adversely affected by CKD, secondary to a reduced oral absorption and glomerular filtration; altered tubular secretion; and reabsorption and changes in intestinal, hepatic, and renal metabolism. In general, drug dosing can be accomplished by multiple methods; however, the most common recommendations are often to reduce the dose or expand the dosing interval, or use both methods simultaneously. Some medications need to be avoided all together in CKD either because of lack of efficacy or increased risk of toxicity. Nevertheless, specific recommendations are available for dosing of certain medications and are an important resource, because most are based on clinical or pharmacokinetic trials. PMID- 15755471 TI - Calcium and phosphorus metabolism in patients who have chronic kidney disease. AB - Disturbances in calcium and phosphorus metabolism are almost invariable consequences of chronic kidney disease (CKD). Because the capacity to regulate calcium and phosphorus metabolism becomes compromised progressively as kidney function declines, calcium and phosphorus homeostasis is disrupted and serum calcium or phosphorus levels are perturbed in many patients with CKD. The level of interest in, and concerns about, abnormalities in calcium and phosphorus metabolism among patients with CKD has increased substantially in recent years. Strategies for clinical management are being revised, and recent recommendations differ substantially from those used previously with a renewed emphasis on safety. PMID- 15755473 TI - Dyslipidemias in patients who have chronic kidney disease. AB - Patients with CKD are at high risk for developing CVD. In fact, most CKD patients have a 10-year risk of coronary heart disease events greater than or equal to 20%, placing them in the highest risk category according to the National Cholesterol Education Program Adult Treatment Panel III guidelines. For this reason, the National Kidney Foundation K/DOQI guidelines for managing dyslipidemia suggest that CKD patients with LDL greater than or equal to 100 mg/dL (2.59 mmol/L) should be treated with diet and a statin. The K/DOQI guidelines also make it clear that the evidence supporting treatment in CKD populations is lacking however, and that additional placebo-controlled trials are needed. In the mean time, the high incidence of CVD makes intensive monitoring and treatment of dyslipidemias in patients with CKD a reasonable clinical approach. PMID- 15755474 TI - Executing change in the management of chronic kidney disease: perspectives on guidelines and practice. AB - In this era of escalating information, costly technology, and an increasing prevalence of chronic complex diseases in an aging population, a systematic approach to execute changes in the care of patients with kidney disease must be developed. Specifically, there is a need to facilitate the translation of research and clinical guidelines into the delivery of quality clinical care. At present in nephrology, there is some knowledge of disease processes, accumulating knowledge about risk factors for progression, and knowledge about how to best deliver care to those with a chronic disease. The current health care environment is not suited to either the care of chronic conditions or to prevention. Information technology should facilitate shared models of care delivery for chronic conditions and allow opportunities to add new knowledge and deliver good care to complex patient groups. To execute change in the management of patients with CKD, medical students, healthcare professionals, and established physicians need to be educated about the prevalence and consequences of CKD. These educational initiatives should be done in the context of cases or specific patients especially for established practitioners, and should be simplified to make analogies to familiar concepts. The concept that CKD is a risk factor for cardiovascular disease, and needs to be managed (as does diabetes and dyslipidemia), should be more clearly articulated. Basic and clinical research in kidney disease has been enhanced by discoveries in vascular biology, diabetes, and cardiology. Much of the clinical research has been limited, however, by lack of clear definition of CKD. The development of the new K/DOQI staging system that defines and classifies the severity of kidney disease may improve the execution of ongoing robust clinical trials. Incorporating this classification system into the clinical practice of all physicians by automatic laboratory reporting of estimates of GFR raises awareness and improves communication between all medical professionals. Collaborative management of CKD patients between different physicians and multidisciplinary teams, in conjunction with the ongoing investigation of treatments and treatment strategies by both clinician and researchers, may well lead to improved outcomes for patients with CKD. Executing change in the management of CKD requires an increased awareness on the part of all clinicians, including nephrologists, regarding the prevalence and importance of the problem of earlier stages of kidney disease. The systematic evaluation of all patients and incorporation of simplified definitions and classification systems should enhance the ability to improve the outcomes of patients with kidney disease irrespective of time of identification. PMID- 15755475 TI - Agonistic effects of the beta-carboline DMCM revealed in GABA(A) receptor gamma 2 subunit F77I point-mutated mice. AB - Affinity of the inverse agonist methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3 carboxylate (DMCM) to the benzodiazepine binding site of the GABA(A) receptor is abolished by a phenylalanine (F) to isoleucine (I) substitution at position 77 of the gamma2 subunit. We tested the effects of DMCM in gene knockin gamma2I77 mice carrying this mutation. Unlike in wild-type mice, DMCM was not able to reverse the GABA-induced reduction of the picrotoxin-sensitive t-butylbicyclophosphoro [35S]thionate ([35S]TBPS) binding to GABA(A) receptor channels in the forebrain sections of gamma2I77 mice. Accordingly, DMCM was not convulsant in the mutant mice even at doses 20-fold higher (60mg/kg, i.p.) than those producing convulsions in wild-type littermate controls (3 mg/kg, i.p.). Neither did DMCM raise the c-Fos levels in gamma2I77 mouse brain. DMCM additionally exhibits a less well described agonistic effect on GABA(A) receptors that is normally masked by its strong inverse agonist effect. DMCM agonistically enhanced the GABA induced reduction in [35S]TBPS binding to the cerebellar granule cell layer in control and mutant mice. In vivo DMCM (20-60 mg/kg i.p.) produced modest anxiolytic-like effects in gamma2I77 mice as assessed by elevated plus maze and staircase tests, but no motor impairment was found in the rotarod test. The results suggest only minor agonistic efficacy for the beta-carboline DMCM. PMID- 15755476 TI - Effects of a novel glutamate transporter blocker, (2S, 3S)-3-[3-[4 (trifluoromethyl)benzoylamino]benzyloxy]aspartate (TFB-TBOA), on activities of hippocampal neurons. AB - Glutamate transporters rapidly take up synaptically released glutamate and maintain the glutamate concentration in the synaptic cleft at a low level. (2S, 3S)-3-[3-[4-(trifluoromethyl)benzoylamino]benzyloxy]aspartate (TFB-TBOA) is a novel glutamate transporter blocker that potently suppresses the activity of glial transporters. TFB-TBOA inhibited synaptically activated transporter currents (STCs) in astrocytes in the stratum radiatum in rat hippocampal slices in a dose-dependent manner with an IC50 of 13 nM, and reduced them to approximately 10% of the control at 100 nM. We investigated the effects of TFB TBOA on glutamatergic synaptic transmission and cell excitability in CA1 pyramidal cells. TFB-TBOA (100 nM) prolonged the decay of N-methyl-D-aspartic acid receptor (NMDAR)-mediated excitatory postsynaptic currents (EPSCs), whereas it prolonged that of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-mediated EPSCs only when the desensitization of AMPARs was reduced by cyclothiazide (CTZ). Furthermore, long-term application of TFB-TBOA induced spontaneous epileptiform discharges with a continuous depolarization shift of membrane potential. These epileptiform activities were mainly attributed to NMDAR activation. Even after pharmacological block of NMDARs, however, TFB TBOA induced similar changes by activating AMPARs in the presence of CTZ. Thus, the continuous uptake of synaptically released glutamate by glial transporters is indispensable for protecting hippocampal neurons from glutamate receptor-mediated hyperexcitabilities. PMID- 15755478 TI - Alterations of striatal NMDA receptor subunits associated with the development of dyskinesia in the MPTP-lesioned primate model of Parkinson's disease. AB - The development of dyskinesias and other motor complications greatly limits the use of levodopa therapy in Parkinson's disease (PD). Studies in rodent models of PD suggest that an important mechanism underlying the development of levodopa related motor complications is alterations in striatal NMDA receptor function. We examined striatal NMDA receptors in the MPTP-lesioned primate model of PD. Quantitative immunoblotting was used to determine the subcellular abundance of NR1, NR2A and NR2B subunits in striata from unlesioned, MPTP-lesioned (parkinsonian) and MPTP-lesioned, levodopa-treated (dyskinetic) macaques. In parkinsonian macaques, NR1 and NR2B subunits in synaptosomal membranes were decreased to 66 +/- 11% and 51.2 +/- 5% of unlesioned levels respectively, while the abundance of NR2A was unaltered. Levodopa treatment eliciting dyskinesia normalized NR1 and NR2B and increased NR2A subunits to 150 +/- 12% of unlesioned levels. No alterations in receptor subunit tyrosine phosphorylation were detected. These results demonstrate that altered synaptic abundance of NMDA receptors with relative enhancement in the abundance of NR2A occurs in primate as well as rodent models of parkinsonism, and that in the macaque model, NR2A subunit abundance is further increased in dyskinesia. These data support the view that alterations in striatal NMDA receptor systems are responsible for adaptive and maladaptive responses to dopamine depletion and replacement in parkinsonism, and highlight the value of subtype selective NMDA antagonists as novel therapeutic approaches for PD. PMID- 15755477 TI - Genetic knockout and pharmacological blockade studies of the 5-HT7 receptor suggest therapeutic potential in depression. AB - The affinity of several antidepressant and antipsychotic drugs for the 5-HT7 receptor and its CNS distribution suggest potential in the treatment of psychiatric diseases. However, there is little direct evidence of receptor function in vivo to support this. We therefore evaluated 5-HT7 receptors as a potential drug target by generating and assessing a 5-HT7 receptor knockout mouse. No difference in assays sensitive to potential psychotic or anxiety states was observed between the 5-HT7 receptor knockout mice and wild type controls. However, in the Porsolt swim test, 5-HT7 receptor knockout mice showed a significant decrease in immobility compared to controls, a phenotype similar to antidepressant treated mice. Intriguingly, treatment of wild types with SB 258719, a selective 5-HT7 receptor antagonist, did not produce a significant decrease in immobility unless animals were tested in the dark (or active) cycle, rather than the light, adding to the body of evidence suggesting a circadian influence on receptor function. Extracellular recordings from hypothalamic slices showed that circadian rhythm phase shifts to 8-OH-DPAT are attenuated in the 5 HT7 receptor KO mice also indicating a role for the receptor in the regulation of circadian rhythms. These pharmacological and genetic knockout studies provide the first direct evidence that 5-HT7 receptor antagonists should be investigated for efficacy in the treatment of depression. PMID- 15755479 TI - KW-6002 protects from MPTP induced dopaminergic toxicity in the mouse. AB - The risk of Parkinson's disease (PD) is associated with a lower intake of caffeine, a non-selective adenosine A2A antagonist. In agreement, genetic or pharmacological inactivation of adenosine A2A receptors in animal models of PD has demonstrated both symptomatic and neuroprotective effects. These findings and the lack of disease modifying therapies have led to intense research on adenosine A2A antagonists as a novel treatment for PD. In the present study the neuroprotective effect of the A2A receptor antagonist KW-6002 was investigated using different models of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice, which induced dopaminergic terminal and or dopaminergic cell loss and inflammation. Treatment with KW-6002 prevented the loss of dopaminergic striatal terminals and nigral cell bodies and inhibited the nigral microglia activation. Our results confirm previous findings that pharmacological inactivation of A2A receptors inhibits MPTP-induced dopaminergic damage at the level of striatum. In addition, we demonstrate for the first time that, after MPTP treatment in mice, an A2A antagonist is neuroprotective, and has anti-inflammatory effects, at the level of the substantia nigra. Thus, our data further support the use of A2A receptor antagonists as a novel neuroprotective therapy for PD. PMID- 15755480 TI - Dopamine inhibits cell growth and cell cycle by blocking ribonucleotide reductase. AB - Dopamine (DA) is a classical neurotransmitter modulating various brain functions by acting on its specific receptors. In addition, DA is a reactive molecule that has been implicated in neurodegeneration, especially in Parkinson's disease. Here we show that DA inhibited cell growth of dopamine transporter transfected cells by intracellularly blocking cell cycle progression. To pinpoint the site of this effect, we measured DNA distribution and 5-bromo-2'-deoxyuridine (BrdU) incorporation, as well as the levels of the key cell cycle proteins. DA increased number of cells with a G1 DNA content, decreased BrdU incorporation and simultaneously increased cyclin A but had no effect on cyclin D2, D3, E, nor on cdk4 and p21. These results narrowed down the DA effect to the beginning of S phase, suggesting inhibition of the ribonucleotide reductase, an enzyme essential for DNA synthesis. Indeed, measurement of enzyme activity in situ revealed that DA, within 1h of addition to cells labelled with [3H]cytidine, strongly reduced the cell content of [3H]2'-deoxycytidine 5'-triphophate. The time course of this DA effect preceded the cell cycle progression. This novel molecular mechanism of intracellular DA action independent of plasmamembrane receptors may be involved in processes controlling the development and survival of brain dopaminergic neurons. PMID- 15755481 TI - Pharmacological reactivity to cocaine in adult rats undernourished at perinatal age: behavioral and neurochemical correlates. AB - The influence of neuronal alterations induced by early undernutrition on the stimulant effect of cocaine was assessed in adult rats submitted to a protein deprivation schedule at perinatal age. To evaluate the sensitization phenomenon induced by repeated cocaine administration, different groups of control (C) and deprived (D) rats received a daily injection of cocaine (5, 10 or 15 mg/kg, i.p.) for 16 days. Behavioral parameters were assessed every two days in an open-field. Dose-response curves obtained with different doses of cocaine used revealed a shift to the left in the locomotor activity curves of D rats compared to controls. Thus, D animals showed a clear behavioral sensitization to the lower dose of cocaine, whereas this phenomenon was only observed in C rats for the higher dose used. To correlate this differential development of sensitization with neurochemical parameters, we assessed extracellular dopamine (DA) levels in nucleus accumbens (core and shell) and in the dorsal caudate-putamen, using a microdialysis technique. A challenge with cocaine in cocaine pre-exposed animals produced a different increase in DA output only in nucleus accumbens "core" of D animals. Comparable DA levels were observed in nucleus accumbens shell and in dorsal caudate-putamen of both groups. These results demonstrate that D rats had a lower threshold developing a progressive behavioral sensitization following repeated cocaine administration, as well as higher responsiveness of the nucleus accumbens (core) expressed by increased DA release. PMID- 15755482 TI - The modulation of Ca2+ and K+ channels but not changes in cAMP signaling contribute to the inhibition of glutamate release by cannabinoid receptors in cerebrocortical nerve terminals. AB - While cannabinoid receptors activate multiple signaling pathways in the brain, it remains unclear what influence the inhibition of adenylylcyclase has on the inhibition of glutamate release. In cerebrocortical nerve terminals, the cannabinoid receptor agonist WIN55,212-2 reduced KCl-evoked glutamate release through a mechanism that restricted the rise of cytoplasmic free Ca2+, but not the changes in plasma membrane depolarization. These effects were consistent with the inhibition of Ca2+ channels. Furthermore, WIN55,212-2 reduced 4-aminopyridine (4AP) evoked glutamate release to a larger extent by modulating the behavior of both Ca2+ and K(+)-channels. The inhibition of 4AP-evoked release was associated with a decrease in cytoplasmic free Ca2+ and in plasma membrane depolarization that was reverted by the potassium channel blocker, tetraethylammonium. Interestingly, the reduction of KCl- and 4AP-evoked release by WIN55,212-2 was independent of adenylylcyclase activity and did not affect cAMP. Forskolin and the beta-adrenergic receptor increase intrasynaptosomal cAMP and promote a PKA dependent tetrodotoxin (TTX)-sensitive increase in the spontaneous release of glutamate. These two responses were reduced by WIN55,212-2. However, the glutamate release induced by Sp-8-Br-cAMPS, which directly activated PKA without affecting cAMP, was also similarly reduced by WIN55,212-2. Hence, we conclude that the inhibition of glutamate release by WIN55,212-2 is unrelated to changes in cAMP and that the inhibition of release that a decrease in cAMP might produce is occluded by the activation of additional pathways such as the inhibition of Ca2+ channels and/or the activation of K(+)-channels that strongly depress glutamate release. PMID- 15755483 TI - Improved posthypoxic recovery in vitro on treatment with drugs used for secondary stroke prevention. AB - Besides aspirin several new drugs for inhibition of platelet aggregation and 3 hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibition are used in secondary stroke prevention. Pharmacology and clinical effects, however, are not fully explained by current understanding of underlying mechanisms. Population spike amplitude (PSAP), an established marker of slice integrity, was measured during hypoxia and recovery thereof in hippocampal slices from control CD1 mice (25-35 g) and animals pretreated in vivo with a single i.p. injection of clopidogrel, ticlopidine, or atorvastatine at different time intervals and dosages. Posthypoxic recovery of PSAP was 20 +/- 35% in control CD1 mice. Upon pretreatment with clopidogrel (1-24 h, 0.5-2 mg/kg body weight) an increase up to 81 +/- 20% (p < 0.01 to control) was observed at 1h interval and 1mg/kg. Application of ticlopidine (1-24 h, 1-4 mg/kg body weight) resulted in an improvement of posthypoxic recovery to 61 +/- 41% (p < 0.05 to control) while administration of atorvastatine (1-24 h, 1-4 mg/kg body weight) caused an increase up to 87 +/- 31% (p < 0.01 to control) at 1h interval and 2 mg/kg. On application of these substances in vitro the NADH autofluorescence spectrum in hippocampal slices is blue-shifted suggesting an alteration of oxidative metabolism. The present data demonstrate a shared neuroprotective effect of agents known to inhibit platelets (acetylsalicylic acid, clopidogrel, and ticlopidine) and HMG-CoA reductase (atorvastatine). The time course of this neuroprotective action in the current experimental study (onset within an hour, duration of several hours in contrast to several days) resembles clinical practice in dosing these substances. We hypothesize that an increase of hypoxic tolerance resulting from mild mitochondrial inhibition by these substances is a principal constituent of the effectiveness of these drugs. PMID- 15755484 TI - Protection of ischemic rat spinal cord white matter: Dual action of KB-R7943 on Na+/Ca2+ exchange and L-type Ca2+ channels. AB - The effect of the Na+/Ca(2+)-exchange inhibitor KB-R7943 was investigated in spinal cord dorsal column ischemia in vitro. Oxygen/glucose deprivation at 37 degrees C for 1 h causes severe injury even in the absence of external Ca2+. KB R7943 was very protective in the presence and absence of external Ca2+ implicating mechanisms in addition to extracellular Ca2+ influx through Na+/Ca(2+)-exchange, such as activation of ryanodine receptors by L-type Ca2+ channels. Indeed, blockade of L-type Ca2+ by nimodipine confers a certain degree of protection of dorsal column against ischemia; combined application of nimodipine and KB-R7943 was not additive suggesting that KB-R7943 may also act on Ca2+ channels. KB-R7943 reduced inward Ba2+ current with IC50 = 7 microM in tsA 201 cells expressing Ca(v)1.2. Moreover, nifedipine and KB-R7943 both reduced depolarization-induced [Ca2+]i increases in forebrain neurons and effects were not additive. Nimodipine or KB-R7943 also reduced ischemic axoplasmic Ca2+ increase, which persisted in 0Ca2+/EGTA perfusate in dorsal column during ischemia. While KB-R7943 cannot be considered to be a specific Na+/Ca2+ exchange inhibitor, its profile makes it a very useful neuroprotectant in dorsal columns by: reducing Ca2+ import through reverse Na+/Ca2+ exchange; reducing influx through L-type Ca2+ channels, and indirectly inhibiting Ca2+ release from the ER through activation of ryanodine receptors. PMID- 15755485 TI - Opposite effects of lithium and valproic acid on trophic factor deprivation induced glycogen synthase kinase-3 activation, c-Jun expression and neuronal cell death. AB - Recent studies demonstrate that lithium and valproic acid (VPA), two commonly used mood-stabilizing drugs, have neuroprotective effects against a variety of insults. Inhibition of the proapoptotic enzyme, glycogen synthase kinase-3 (GSK 3), has been suggested to be the mechanism of action of neuroprotection for both drugs. In this study, we tested if lithium and VPA could protect cultured cerebellar granule neurons (CGNs) from GSK-3-mediated apoptosis induced by trophic factor withdrawal (serum/potassium deprivation). Both lithium and indirubin, a specific GSK-3 inhibitor, protected CGNs in a dose-dependent manner. In contrast, VPA did not provide any neuroprotection and even potentiated cell death. Immunoblot analysis revealed that lithium inhibited the trophic factor deprivation-induced activation of GSK-3 as well as the in vivo phosphorylation of the microtubule-associated protein Tau on Ser199, a specific target site for GSK 3. Under these same experimental conditions, however, VPA neither inhibited GSK-3 activation nor hindered GSK-3 mediated Tau phosphorylation. Furthermore, in accordance with their effects on neuronal survival, lithium prevented the induction of c-Jun expression in trophic factor-deprived CGNs, whereas VPA potentiated it. Collectively, these results show that VPA is not a universal inhibitor of neuronal GSK-3, and that instead of being neuroprotective, VPA can even exacerbate neuronal death under some conditions. PMID- 15755486 TI - Neurosteroid paradoxical enhancement of paired-pulse inhibition through paired pulse facilitation of inhibitory circuits in dentate granule cells. AB - Neurosteroids are produced in the brain independently of peripheral endocrine glands to act locally in the nervous system. They exert potent promnesic effects and play significant roles in mental health-related disorders. In part, neurosteroids act by affecting ligand-gated ion channels and metabotropic receptors through rapid non-genomic processes. We have previously demonstrated that neurosteroids also affect synaptic transmission presynaptically in the CA1 region of the hippocampus. Here we describe the effects of the most abundant neurosteroid in the rodent brain, pregnenolone sulfate (PregS), on signal processing in the dentate subfield of the hippocampus. We show that PregS acts presynaptically at low concentrations (300 nM) to enhance paired-pulse facilitation (PPF) in perforant pathway terminals on dentate granule cells. Similar effects were found with two steroid sulfatase inhibitors demonstrating a potential contribution of endogenous steroids to dentate synaptic plasticity. This enhanced presynaptic facilitation paradoxically increases paired-pulse inhibition (PPI) at short interpulse intervals. Based on these data, a model of dentate gyrus circuit interactions is proposed for the presynaptic action of PregS on the filtering dynamics of the dentate subfield at frequencies similar to those of the endogenous signals from the entorhinal cortex. These modeling studies are consistent with experimental measurements demonstrating positive modulation by PregS at low frequencies and negative modulation at high frequencies. These studies show an important role for the presynaptic action of neurosteroids in modulating input signals to the hippocampus. PMID- 15755487 TI - Non-adrenergic inhibition at prejunctional sites by agmatine of purinergic vasoconstriction in rabbit saphenous artery. AB - We investigated the effects of agmatine, clonidine, xylazine and moxonidine on the purinergic vasoconstriction induced by electrical stimulation in the rabbit isolated saphenous artery without endothelium. Transmural electrical stimulations induced reproducible responses in the arterial preparations, which were abolished by tetrodotoxin at 0.1 microM or pyridoxal-phosphate-6-azophenyl-2',4' disulphonic acid tetrasodium salt (PPADS, 30 microM), but were not affected by 1 microM prazosin. Clonidine, xylazine and moxonidine induced transient and concentration-independent vasoconstriction, with threshold concentrations of 1, 3 and 30 microM, respectively. Agmatine, in contrast, did not produce any vascular response even at 1 mM. Lower concentrations of clonidine, xylazine and moxonidine (0.01-0.3 microM) concentration-dependently decreased vasoconstrictor responses to electrical stimulation, whereas agmatine (0.1-1 mM) induced an inhibitory followed by a facilitatory effect on electrically evoked responses. Agmatine, clonidine and moxonidine but not xylazine significantly enhanced the vasoconstriction elicited by 1 mM ATP. The concentration-response curve for NA was shifted to the left slightly by 1 mM agmatine, but not affected by 0.3 microM of other three agonists. Phenoxybenzamine did not affect the vasoconstrictive responses to 1mM ATP and to electrical stimulations, but abolished those to NA. Agmatine at 1mM evoked only an inhibitory effect on electrical stimulation induced vasoconstriction in the preparation pretreated with phenoxybenzamine, and the inhibitory action was enhanced to 38.6% from the control value (without treatment with phenoxybenzamine) of 22.5%. The non-imidazoline compound xylazine at 0.3 microM lost its inhibitory effect on the neurogenic vasoconstriction in the presence of phenoxybenzamine. In conclusion, agmatine produces a biphasic effect on the purinergic vasoconstriction induced by sympathetic nerve stimulation in the rabbit isolated saphenous artery. The monophasic inhibition of agmatine in the artery treated with phenoxybenzamine is due to an alpha adrenoceptor-independent mechanism at prejunctional sites, and the potentiation effect of agmatine is mainly dependent on its enhancement of vasoconstriction at postjunctional sites. PMID- 15755488 TI - Comparison of responses of ventral posterolateral and posterior complex thalamic neurons in naive rats and rats with hindpaw inflammation: mu-opioid receptor mediated inhibitions. AB - The aim of the present study was to compare the effects of morphine on thalamic neuronal responses in naive rats and rats with carrageenan-induced hindpaw inflammation. Multiple single unit ventral posterolateral (VPL) and posterior complex (Po) activity was recorded and mechanically- (7 g, 14 g, 21 g, 60 g and 80 g) evoked responses of VPL and Po neurones were measured in naive rats and rats with carrageenan (100 microl, 2%)-induced hindpaw inflammation. Effects of systemic (0.5 mg kg(-1)) and intra-thalamic (66 microM, 250 nL) morphine on neuronal responses were determined. Mechanically-evoked (60 g) nociceptive responses of VPL neurones were significantly larger in inflamed rats (29 +/- 4 spikes s(-1)) compared to naive rats (19 +/- 2 spikes s(-1), P < 0.05). Systemic morphine inhibited 7 g-evoked responses of VPL neurones in inflamed (24 +/- 8% control, P < 0.01), but not in naive rats (123 +/- 3% control). Frank noxious evoked responses of VPL neurones in inflamed rats were less sensitive to the effects of systemic and intra-thalamic morphine, compared to naive rats (P < 0.05 for both). These data provide evidence for altered evoked responses of neurones at the level of VPL, but not at Po, during hindpaw inflammation and suggest that thalamic sites of action contribute to the effects of systemic morphine. PMID- 15755489 TI - The Symposia on Calcium Binding Proteins and Calcium Function in Health and Disease: an historical account, and an appraisal of their role in spreading the calcium message. PMID- 15755490 TI - Optical single-channel recording by imaging Ca2+ flux through individual ion channels: theoretical considerations and limits to resolution. AB - Recent developments in microscopy and fluorescent indicators now make it possible to monitor the activity and localization of membrane ion channels by imaging Ca(2+) flux through individual channels. Such optical approaches have advantages over electrophysiological single-channel techniques in that they are less invasive, provide spatial information and can simultaneously and independently monitor hundreds of channels. However, their kinetic resolution does not yet approach that of patch-clamp recordings. To help understand the processes that determine the temporal resolution and noise level of single-channel Ca(2+) fluorescence signals (SCCaFTs), we simulated the microdomains of Ca(2+) ions and Ca(2+)-bound indicator dye that exist around the mouth of an open channel. Further, as an aid to development of improved optical techniques, we modeled the dependence of the amplitude and kinetics of SCCaFTs on parameters such as the imaging volume, the indicator concentration, affinity and mobility, and the presence of endogenous and exogenous Ca(2+) buffers. The results indicate that under optimal conditions, including the use of confocal or total-internal reflection microscopy to image from sub-femtolitre volumes, SCCaFTs should resolve channel openings as brief as 1ms with a signal-to-noise ratio >10. PMID- 15755491 TI - Expression of multiple CaV1.2 transcripts in rat tissues mediated by different promoters. AB - The expression of two different transcripts for Ca(V)1.2 in rat tissues mirrors that which has previously been described for human tissue, in that expression of transcripts expressing exon 1a is predominant only in heart, whereas expression of transcripts expressing exon 1b is greater in smooth muscle rich tissues such as aorta and intestine. Transcripts expressing exon 1b also predominate in brain and in diaphragm. Western blots indicate that the N-terminus coded for by exon 1b is present in much of the protein in all these tissues except heart. The promoter just upstream of exon 1b has been cloned, sequenced and utilized to drive expression of luciferase in smooth muscle A7r5 cells, cardiac HL-1 cells, skeletal muscle L6 cells and neuronal PC12 cells. The nucleotide sequence of the promoter exhibits 80% identity with the equivalent promoter previously identified in humans and 94% identity with the sequence of the equivalent region of the mouse genome. Evidence in favor of still another promoter upstream of exon 2 has been uncovered. PMID- 15755492 TI - Evidence for signaling via gap junctions from smooth muscle to endothelial cells in rat mesenteric arteries: possible implication of a second messenger. AB - We investigated heterocellular communication in rat mesenteric arterial strips at the cellular level using confocal microscopy. To visualize Ca(2+) changes in different cell populations, smooth muscle cells (SMCs) were loaded with Fluo-4 and endothelial cells (ECs) with Fura red. SMC contraction was stimulated using high K(+) solution and Phenylephrine. Depending on vasoconstrictor concentration, intracellular Ca(2+) concentration ([Ca(2+)](i)) increased in a subpopulation of ECs 5-11s after a [Ca(2+)](i) rise was observed in adjacent SMCs. This time interval suggests chemical coupling between SMCs and ECs via gap junctions. As potential chemical mediators we investigated Ca(2+) or inositol 1,4,5 trisphosphate (IP(3)). First, phospholipase C inhibitor U-73122 was added to prevent IP(3) production in response to the [Ca(2+)](i) increase in SMCs. In high K(+) solution, all SMCs presented global and synchronous [Ca(2+)](i) increase, but no [Ca(2+)](i) variations were detected in ECs. Second, 2 aminoethoxydiphenylborate, an inhibitor of IP(3)-induced Ca(2+) release, reduced the number of flashing ECs by 75+/-3% (n = 6). The number of flashing ECs was similarly reduced by adding the gap junction uncoupler palmitoleic acid. Thus, our results suggest a heterocellular communication through gap junctions from SMCs to ECs by diffusion, probably of IP(3). PMID- 15755493 TI - Modeling stochastic Ca2+ release from a cluster of IP3-sensitive receptors. AB - We focused our attention on Ca(2+) release from the endoplasmic reticulum through a cluster of inositol(1,4,5)-trisphosphate (IP(3)) receptor channels. The random opening and closing of these receptors introduce stochastic effects that have been observed experimentally. Here, we present a stochastic version of Othmer Tang model (OTM) for IP(3) receptor clusters. We address the average behavior of the channels in response to IP(3) stimuli. In our stochastic simulation we found that the fraction of open channels versus [IP(3)] follows a Hill curve, whose associate Hill coefficient increases when intracellular Ca(2+) level increase. This finding suggests that feedback from cytosolic Ca(2+) plays a key role in the channel response to IP(3). We also study several aspects of the stochastic properties of Ca(2+) release and we compare with experimental observations. PMID- 15755494 TI - Rearrangement of the close contact between the mitochondria and the sarcoplasmic reticulum in airway smooth muscle. AB - The mitochondria and the sarcoplasmic reticulum (SR) are two major intracellular calcium-storing organelles that exhibit close functional interaction with each other. Close spatial association is believed to be important for their functional interaction. In this study, we have characterized the spatial relationship between the SR and the mitochondria in porcine tracheal smooth muscle cells (TSMC) under different conditions. By examining the cross-section of unstimulated TSMC with electron microscopy, we found that 99.4 +/- 0.5% of the mitochondria seen on random cross-sections were situated within 30 nm of the SR and that 82.2 +/- 6.7% of the mitochondria were completely enveloped by the SR network. Overall, 48.0 +/- 3.5% of the mitochondrial outer membrane was within 30 nm with the SR. After stimulation of the TSMC with acetylcholine (ACh) or 80 mM [K(+)] solution 97.0 +/- 2.1% and 98.6 +/- 1.4% of the mitochondria observed were situated within 30 nm of the SR, respectively. However, the proportion of the mitochondria that was completely enveloped by the SR was significantly reduced to 12.2 +/- 5.9% in ACh-stimulated cells and 9.7 +/- 6.6% in 80 mM [K(+)] stimulated cells. The percentage of mitochondrial membrane closely associated with the SR was correspondingly lower at 10.1 +/- 1.0% during ACh stimulation and 10.8 +/- 0.9% during 80 mM [K(+)] stimulation. During smooth muscle cell stimulation, the SR appears to unwrap from the mitochondria and extend into the cytoplasm while maintaining close contact with the mitochondria over a smaller area. Such static and dynamic components of the close spatial association between the mitochondria and the SR may serve as a structural basis for the selective and efficient Ca(2+) trafficking between the two organelles in TSMC. PMID- 15755495 TI - Elimination of environmental sensitivity in a cameleon FRET-based calcium sensor via replacement of the acceptor with Venus. AB - Genetically encoded sensors are becoming a powerful tool for investigating cellular signaling pathways and, potentially, signaling in vivo. Many sensors use changes in fluorescence resonance energy transfer (FRET) between donor and acceptor variants of GFP separated by a ligand binding domain sensitive to a particular signaling pathway. Accurate measurements require that sensors be insensitive to extraneous intracellular environmental factors. We have found that the responsiveness of the Ca(2+) sensor, cameleon YC6.1, varies linearly with the resting YFP/CFP emission ratio in the cell. However, cells expressing responsive or non-responsive sensor can easily be segregated by determining a resting YFP/CFP ratio cutoff for the sensor. This environmental sensitivity has been eliminated by replacing EYFP with Venus to produce a new cameleon we have designated VC6.1. Measurements show that VC6.1 has a greater dynamic range than YC6.1 and better environmental resistance. We also show that YC6.1 is inactivated by persistent activation of the IP(3) pathway following expression of constitutively active G(q), while VC6.1 is not. The stability of VC6.1 may make it well suited to studies utilizing mixed cell populations such as those encountered in vivo. PMID- 15755496 TI - Two-photon microscopic analysis of acetylcholine-induced mucus secretion in guinea pig nasal glands. AB - The spatiotemporal changes in intracellular free Ca(2+) concentration ([Ca(2+)](i)) as well as fluid secretion and exocytosis induced by acetylcholine (ACh) in intact acini of guinea pig nasal glands were investigated by two-photon excitation imaging. Cross-sectional images of acini loaded with the fluorescent Ca(2+) indicator fura-2 revealed that the ACh-evoked increase in [Ca(2+)](i) was immediate and spread from the apical region (the secretory pole) of acinar cells to the basal region. Immersion of acini in a solution containing a fluorescent polar tracer, sulforhodamine B (SRB), revealed that fluid secretion, detected as a rapid disappearance of SRB fluorescence from the extracellular space, occurred exclusively in the luminal region and was accompanied by a reduction in acinar cell volume. Individual exocytic events were also visualized with SRB as the formation of Omega-shaped profiles at the apical membrane. In contrast to the rapidity of fluid secretion, exocytosis of secretory granules occurred with a delay of approximately 70s relative to the increase in [Ca(2+)](i). Exocytic events also occurred deep within the cytoplasm in a sequential manner with the latency of secondary exocytosis being greatly reduced compared with that of primary exocytosis. The delay in sequential compound exocytosis relative to fluid secretion may be important for release of the viscous contents of secretory granules into the nasal cavity. PMID- 15755497 TI - Rapid Ca2+-dependent increase in oxygen consumption by mitochondria in single mammalian central neurons. AB - Oxygen consumption increases within a fraction of a second after the onset of neuronal activity, a phenomenon referred to as the "initial dip" in functional imaging studies of the living brain. The cellular mechanism that underlies this rapid increase in oxygen consumption has remained unclear, however. We have now used two-photon excitation imaging to characterize rapid activity-dependent mitochondrial responses in single neurons. This approach allowed simultaneous multicolor imaging of individual mitochondria in single mouse Purkinje neurons in culture. Mitochondrial depolarization was induced immediately when the cytosolic free Ca(2+) concentration ([Ca(2+)](i)) exceeded 15 microM and was associated with oxidation of mitochondrial NAD(P)H, suggesting that Ca(2+)-induced mitochondrial depolarization mediated by the Ca(2+) uniporter directly facilitated oxidation of NAD(P)H. With the use of a miniature oxygen electrode, we detected a burst of oxygen consumption within 0.2s after the onset of cell depolarization in single Purkinje neurons, and this rapid increase in oxygen consumption was dependent on the increase in [Ca(2+)](i). We have thus demonstrated a rapid Ca(2+)-dependent consumption of oxygen that is mediated by mitochondrial depolarization in mammalian central neurons. This process might function as a rapid feed-forward mechanism in homeostatic control of the cytosolic ATP concentration. PMID- 15755498 TI - A role for Wnt signaling in adult T-cell leukemia? PMID- 15755499 TI - Letting the CAThelicidin out of the bag, as a therapeutic modulator of the adaptive immune system. PMID- 15755500 TI - Anagrelide: analysis of long-term efficacy, safety and leukemogenic potential in myeloproliferative disorders. AB - Appropriate treatment for nonreactive thrombocytosis resulting from a myeloproliferative disorder (MPD) is surrounded by controversy. Although few doubt the association of thrombocytosis with increased risk for life-threatening events such as thrombosis or hemorrhage, or the association between clonal myeloproliferation and the progression to acute leukemia or myelofibrosis, controversy exists regarding the timing and nature of appropriate therapeutic intervention. Studies have shown that treatment with myelosuppressive agents such as chlorambucil, busulfan, radiophosphorus (32P), and hydroxyurea reduces the platelet count. However, investigators have also identified an increased risk of drug-related leukemic transformation. An ideal cytoreductive treatment for long term use should minimize thrombosis and avoid long-term complications, especially acute leukemia (AL). Anagrelide, an imidazoquinolin, inhibits megakaryopoiesis and more selectively reduces platelet production in humans. A retrospective analysis of an open-label, multicenter, international trial reviewing 3660 anagrelide-treated patients was performed to assess efficacy and long-term safety, specifically potential for increased leukemogenicity. The study included MPD patients with thrombocytosis diagnosed according to Polycythemia Vera Study Group (PVSG) criteria. Of all patients enrolled, 81% had previously received other myelosuppressive agents; of these, 33% were transferred from the original agent to anagrelide due to toxicity and 31% were transferred because of poor platelet control. Over 45% of patients were symptomatic due to thrombocythemia, most commonly vascular sequelae (25%). Dosage was titrated to achieve a platelet count < 600 x 10(9) L(-1) and ideally between 130 and 450 x 10(9) L(-1). The safety cohort of 3660 patients, including 2251 with essential thrombocythemia (ET), 462 with polycythemia vera (PV), and 947 with chronic myeloid leukemia (CML) and other MPDs, was analyzed to establish the incidence of leukemic transformation in patients with ET and PV. From the Safety Population, 12.8% (467/3660) of patients were treated with anagrelide as the sole cytoreductive agent (naive patients). Acute leukemia/myelodysplasia developed in 2.1% of ET patients (47/2251) with a maximum follow-up of 7.1 years. Of the PV patients, 2.8% developed acute leukemia/myelodysplastic syndrome (13/462), with a maximum follow-up of 7.0 years. ET and PV patients who transformed to AL had all been previously exposed to other cytotoxics; there were no ET or PV patients in the study who transformed to AL exposed solely to anagrelide. With maximum follow-up over 7 years, anagrelide achieved platelet control in over 75% of MPD patients and did not increase the conversion to acute leukemia during the treatment duration analyzed. Longer follow-up is required to confirm these important observations regarding the long-term safety of anagrelide. PMID- 15755501 TI - Chronic disseminated candidiasis in patients with acute leukemia: emphasis on diagnostic definition and treatment. AB - BACKGROUND: Chronic disseminated candidiasis (CDC) is a form of invasive fungal infection that occurs most commonly in patients with acute leukemia treated with chemotherapy. Recent studies have provided evidence for diagnostic alternatives to invasive procedures and more therapeutic options for the management of this complication. In order to put diagnostic criteria and methodological approach to the disease into the perspective of developing strategies for therapy, all relevant studies published in the English literature over the last 30 years were examined. MATERIALS AND METHODS: The English-language articles located through MEDLINE (1966 to present) and from selected bibliographies. RESULTS: There is increased recognition of CDC as complication of treatment with chemotherapy in patients with acute leukemia. Liver biopsy may not always be revealing or feasible to perform in some patients. Among the imaging modalities, magnetic resonance imaging has obtained preeminence as a non-invasive tool for the diagnosis of hepatosplenic fungal infections. Administration of amphotericin B (Amp B) in relatively large cumulative doses is needed to ensure appropriate control of the infection and prevention of future relapse. Patients intolerant of, or refractory to conventional Amp B have been successfully salvaged using fluconazole or lipid formulations of Amp B. A constellation of clinical, laboratory and radiologic parameters should be used to determine response and efficacy of therapy. There is sufficient evidence to support the safety and feasibility of continuing chemotherapy for acute leukemia in conjunction with antifungal treatment in patients diagnosed with CDC. CONCLUSION: The development of CDC in patients with acute leukemia does not preclude further chemotherapy or constitute contraindication for bone marrow transplantation. Knowledge of the course and pattern of evolution of the disease and adopting aggressive therapeutic approach will likely reduce the morbidity and mortality from this complication. PMID- 15755502 TI - Serological identification of immunogenic antigens in acute monocytic leukemia. AB - In order to improve disease-free survival and potentially a cure, it is necessary to identify more potent leukemia antigen. Here, we defined the acute monocytic leukemia-associated antigen (LAA) recognized by the humoral immune system for the first time. We have applied the method of serologic analysis of recombinant cDNA expression library (SEREX) on acute monocytic leukemia (FAB M5), followed by DNA sequencing and analyzing of positive clones. Then, the reactivity of normal and other leukemia sera with positive clones were performed. Thirty-five distinct novel antigens reactive with autologous IgG were identified by SEREX analysis on an acute monocytic leukemia patient and were characterized according to cDNA sequence and the reactivity with allogeneic sera. Twenty of the 35 antigens identified in this study were recognized by IgG antibodies in normal sera, and the remaining 15 were recognized exclusively by sera from allogeneic leukemia patients but not by normal donor sera, suggested that the immune response to these 15 antigens are leukemia related. The 15 immunogenic antigens detected by immune responses in the autologous host facilitate the identification of epitopes recognized by antigen-specific cytotoxic T lymphocytes (CTL) and are potential candidates for diagnosis and immunotherapy in acute myeloid leukemia (AML). PMID- 15755503 TI - Methylation analysis of the cell cycle control genes in myelofibrosis with myeloid metaplasia. AB - Promoter hypermethylation represents a primary mechanism in the inactivation of tumor suppressor genes during tumorigenesis. We analyzed the promoter methylation status of eight tumor-associated genes (p14 ARF, p15 INK4B, p16 INK4A, Rb, hMLH1, hMSH2, APC, and DAPK) in 30 patients with myelofibrosis with myeloid metaplasia (MMM) by methylation specific PCR. The study showed no hypermethylation of the promoters of p16(INK4A), Rb, hMLH1, hMSH2, APC, and DAPK genes. The p14 ARF, p15 INK4B promoters were hypermethylated in only one patient each. This study indicates that, although methylation of these genes is important in other cancers, it is rare in MMM and causation of this disease should be focused elsewhere. PMID- 15755504 TI - Identification of various MLL gene aberrations that lead to MLL gene mutation in patients with acute lymphoblastic leukemia (ALL) and infants with acute leukemia. AB - Studies were done to investigate MLL gene aberrations using Conventional Cytogenetics, Southern blotting as well as FISH using a panel of probes on 218 cases which included 206 cases of pediatric/young adult ALL and 12 cases of infantile acute leukemias from Tata Memorial Hospital, India. The incidence of MLL gene rearrangements in acute lymphoblastic leukemia (ALL) was 9.4% which included infants as well as pediatric/young adults. In infantile group which included ALL as well as AML cases, MLL gene rearrangement was very common (75% frequency). Application of metaphase-FISH helped unravel MLL rearrangements not only as a result of translocations but also inversions, insertions, partial deletion, duplications, partial duplication-->self-fusion. Besides age, MLL gene rearrangements showed significant association with hyperleukocytosis, peripheral blood blast percentage and early Pre-B phenotype. Clinical outcome of patients with MLL gene rearrangements revealed unfavorable prognosis. PMID- 15755505 TI - Lovastatin alters the isoprenoid biosynthetic pathway in acute myelogenous leukemia cells in vivo. AB - Lovastatin, a competitive inhibitor of hydroxymethylglutaryl coenzyme A reductase (HMGR), is used therapeutically to lower plasma cholesterol levels and has garnered attention for its cytotoxic effects in leukemia cells. In this study, escalating doses of lovastatin were administered to nine patients with acute myelogenous leukemia. Peripheral blood leukemia cells were drawn pre- and post lovastatin dosing. Plasma lovastatin bioactivity ranged up to 234 nM lovastatin equivalents. Our results show that in vivo lovastatin, at up to 200 mg/dose, induces an increased activity of leukemia cell HMGR and alters leukemia cell proliferation without discernibly altering Ras processing. PMID- 15755506 TI - LL-37 enhances adaptive antitumor immune response in a murine model when genetically fused with M-CSFR (J6-1) DNA vaccine. AB - DNA vaccine against M-CSFR(J6-1) (macrophage colony-stimulating factor receptor cloned from the J6-1 leukemic cell line) has shown both protective and therapeutic effects. In this study, to explore the adjuvant effects of LL-37 to M CSFR(J6-1) DNA vaccines, we constructed genetically fused vaccines encoding M CSFR(J6-1) and LL-37(pF). After immunizing BALB/c mice, specific humoral and cellular immune responses were detected. Compared with pR (encoding the extracellular region of M-CSFR(J6-1)), pF was more effective in inducing humoral and cytotoxic immune response, prolonging survival of mice challenged with SP2/0 CSFR(J6-1) tumor cells, and inducing IFN-gamma and IL-4 release by splenocytes. In this study, we also constructed pLL37 (encoding the mature LL-37) and coadministrated pLL37 and pR to see whether the genetic fusion was necessary. We found that compared with pR alone, pLL37+pR could not prolong survival of mice challenged with SP2/0-CSFR(J6-1) tumor cells. Our results suggest that when genetically fused with M-CSFR(J6-1), LL-37 could enhance adaptive immune response against M-CSFR(J6-1) in a murine model challenged with tumor cells bearing M CSFR(J6-1). PMID- 15755507 TI - Inhibition of HHV-8/KSHV infected primary effusion lymphomas in NOD/SCID mice by azidothymidine and interferon-alpha. AB - Kaposi's sarcoma-associated herpesvirus/human herpesvirus type-8 (KSHV/HHV-8) is associated with primary effusion lymphomas (PEL), a rare form of B-cell lymphoma. PEL cell lines infected with HHV-8, but negative for Epstein-Barr virus (EBV), were analyzed for their tumorigenic potential in a small animal model system. Inoculation of PEL cell lines into non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice results in efficient engraftment and tumorigenesis in vivo. PEL-engrafted NOD/SCID (PEL/SCID) mice displayed malignant ascites development with notable abdominal distension, consistent with the clinical manifestations of PEL in humans. Azidothymidine (AZT, zidovudine) and interferon-alpha (IFN-alpha) induce apoptosis in HHV-8+/EBV- PEL cells in culture, by induction of a tumor necrosis factor-related apoptosis inducing ligand (TRAIL) mediated suicide program and has been proposed as a therapy for herpesvirus-associated lymphomas. Daily injection of AZT and IFN-alpha significantly increased mean survival time (MST) of PEL/SCID mice suggesting that induction of apoptosis in PEL cells in vivo may be exploited as an effective relatively non-toxic therapy targeting HHV-8 infected PEL. These data demonstrate that the PEL/SCID mouse is an important preclinical model to characterize efficacy and anti-tumor mechanisms of new therapeutic targets in vivo and will be useful in the design and testing of agents in viral lymphoproliferative diseases. PMID- 15755508 TI - p15 mRNA expression detected by real-time quantitative reverse transcriptase polymerase chain reaction correlates with the methylation density of the gene in adult acute leukemia. AB - Cyclin-dependent kinase inhibitor p15 is frequently inactivated by either methylation or deletion in patients with acute leukemia. To examine pathologic and clinical significance of the p15 gene inactivation, we established a quantitative assay of p15 mRNA expression in the bone marrow cells by real-time quantitative reverse transcriptase-polymerase chain reaction. p15 mRNA expression in 14 patients with precursor B-cell acute lymphoblastic leukemia (PBC-ALL) well correlated with status of deletion and methylation in the p15 gene analyzed by Southern blotting. Furthermore, two patients with PBC-ALL and 11 acute myeloblastic leukemia (AML) were quantitatively examined for p15 gene methylation using bisulfite genomic sequencing. The data showed that p15 mRNA expression significantly correlated with the CpG island methylation density. Among 108 AML patients, p15 mRNA expression was significantly lower in the myeloid lineage (M1, M2, M3) than the monocytic lineage (M4, M5) (P = 0.0019). Above all, the majority of M3 patients showed low p15 expression compared with M1 and M2 patients (P = 0.029). These observations suggest that quantitative analysis of p15 mRNA will be useful to evaluate transcriptional repression of the p15 gene caused by various degrees of methylation. PMID- 15755509 TI - Cytokinetics and mechanism of action of AKO4: a novel nitrogen mustard targeted to bcr-abl. AB - The "combi-targeting" concept seeks to design molecules to not only block tyrosine kinase (TK) activity but also to induce DNA damage. Here we design AK04, a molecule that combines the pharmacophore chlorambucil with that of STI-571 (Gleevec). The results showed that although a less potent abl TK inhibitor than STI571, AK04 was capable of significantly blocking bcr-abl phosphorylation not only in a purified abl assay but also in the bcr-abl+ K562 cells. In contrast to STI571 and like chlorambucil, it induced a dose-dependent increase in DNA damage in these cells. More importantly, AK04 was 12-32-fold more potent than chlorambucil in all bcr-abl+ cells of our cell panel. In the isogenic human megakaryocytic Mo7e and Mo7/bcr-abl cells, AK04 selectively killed the bcr-abl transfectants. Flow cytometry revealed that despite being a five-fold less potent inhibitor of bcr-abl than STI-571, it induced a significant dose-dependent increase in levels of cell death by apoptosis in KU812 cells 24 h post-treatment. Under these conditions, chlorambucil did not induce any significant level of apoptosis. These results suggest that AK04 is a nitrogen mustard with binary bcr abl/DNA targeting effects, a property that may account for its superior potency when compared with the classical mustard chlorambucil. PMID- 15755510 TI - Dietary bioflavonoids induce apoptosis in human leukemia cells. AB - Dietary bioflavonoids are secondary metabolites of plants that are known to have a variety of bio-effects, including anti-cancer activity. In this study, we examined the effects of flavonoids on the growth of human leukemia cells and found that certain flavonoids induce apoptosis in a variety of human leukemia cells. The apoptosis induced by bioflavonoids was dose-dependent and was accompanied by a disruption of the mitochondrial transmembrane potential and the activation of caspase. Our data suggests that dietary bioflavonoids may be useful chemotherapeutic reagents for leukemia patients. PMID- 15755511 TI - Immune therapy of chronic myelogenous leukemia. PMID- 15755512 TI - Bortezomib in combination with dexamethasone for relapsed multiple myeloma. AB - Fifteen patients with advanced multiple myeloma were scheduled to receive bortezomib 1.3 mg/m2 IV days 1, 4, 8, and 11 every 3 weeks for eight cycles in combination with dexamethasone. One patient (7%) achieved a complete response, 10 (67%) a partial response, and one (7%) a minor response (MR) resulting in an overall response rate (> or = MR) of 80% (9/9 with > or = 2nd untreated and 3/6 with refractory relapse). Responses occurred after a median of 3 weeks and were independent of conventional prognostic parameters including deletion of chromosome 13. Adverse events, mainly myelosuppression, neuropathy and fatigue, were manageable. PMID- 15755513 TI - PET scan-positive cat scratch disease in a patient with T cell lymphoblastic lymphoma. AB - In patients who have history of lymphoma, a positive positron emission tomography (PET) scan is frequently considered as good evidence for relapse and/or persistent disease. Thus, lymph node biopsy is not always done to confirm the diagnosis of relapse or refractory lymphoma before a patient is subjected to further chemotherapy. We report a case of patient with history of T cell lymphoblastic lymphoma who presented again with inguinal lymphadenopathy and positive study on positron emission tomography suggestive of lymphoma relapse. This was pathologically proven to be cat scratch disease. This case suggests that in the immunocompromised patients who had history of lymphoma, infectious etiology should be ruled out for PET scan-positive lymphadenopathy. PMID- 15755514 TI - Lack of association of CYP3A4-V polymorphism with the risk of treatment-related leukemia. PMID- 15755515 TI - Decreased GABA(A) receptor expression in the seizure-prone fragile X mouse. AB - The fragile X mental retardation syndrome is due to the transcriptional silence of the fragile X gene, FMR1, and to the resulting loss of the FMR1 product, FMRP. The pathogenesis of the syndrome, however, is not understood. Increased prevalence of childhood seizures is a feature of the fragile X syndrome and increased seizure susceptibility is seen in the fragile X knock out mouse model for this disorder. To investigate the increased seizure susceptibility, we examined GABA(A) receptor expression in the FVB/N fragile X mouse. Western blot analysis revealed that expression of the GABA(A) receptor beta subunit (GABA(A) beta), which is required for receptor function, was reduced in the cortex, hippocampus, diencephalon and brainstem in adult male fragile X mice. Immunohistochemical analysis of brain sections indicated a reduction in GABA(A) beta immunoreactivity. We also found increased expression of glutamic acid decarboxylase, the enzyme responsible for GABA synthesis, in the same regions that showed GABA(A) beta reduction. These results indicate that the absence of Fmrp leads to GABAergic system alterations that could account for the increased seizure susceptibility of the fragile X mouse. These alterations may also be relevant to the seizures and the abnormal behaviors in the human syndrome. PMID- 15755516 TI - The inhibitor of I kappa B alpha phosphorylation BAY 11-7082 prevents NMDA neurotoxicity in mouse hippocampal slices. AB - NF-kappaB is a nuclear transcription factor involved in the control of fundamental cellular functions including cell survival. Among the many target genes of this factor, both pro- and anti-apoptotic genes have been described. To evaluate the contribution of NF-kappaB activation to excitotoxic insult, we analysed the effect of IkappaBalpha (IkappaBalpha) phosphorylation blockade on glutamate-induced toxicity in adult mouse hippocampal slices. By using immunocytochemical and EMSA techniques, we found that (i) acute exposure of hippocampal slices to NMDA induced nuclear translocation of NF-kappaB, (ii) NMDA mediated activation of NF-kappaB was prevented by BAY 11-7082, an inhibitor of IkappaBalpha phosphorylation and degradation, and (iii) BAY 11-7082-mediated inhibition of NF-kappaB activation was associated with neuroprotection. PMID- 15755517 TI - Dopamine modulates synaptic activity in the optic lobes of cuttlefish, Sepia officinalis. AB - The effects of dopamine on spontaneous excitatory postsynaptic currents (sEPSCs) and inhibitory postsynaptic currents (sIPSCs) in three different classes of neurones within the optic lobe of cuttlefish were investigated using whole-cell voltage clamp techniques in a slice preparation. The neuronal types were centrifugal and amacrine neurones, located in the inner granular cell layer, and medullar interneurones, located within the central medulla of the optic lobes. The results demonstrate that bath application of dopamine (50 microM) reversibly reduced both the frequency and amplitude of sEPSCs and of sIPSCs in these optic lobe neurones. The inhibitory effects of DA were dose-dependent and neither D1- nor D2-like receptors appear to be implicated, but probably D4-like receptors are involved in these actions. By pre-applying tetrodotoxin (TTX, 0.5 microM), to block action potential-dependent EPSCs and IPSCs, it is shown that dopamine has no effect on the amplitude, frequency or decay time constant of the mEPSCs or mIPSCs. The results are the first to identify a specific physiological action of dopamine on cephalopod brain activity, they indicate that this effect is probably presynaptic to the specific classes of cells recorded from, and they provide information on the pharmacological profile of the receptors involved. The widespread inhibitory effect of dopamine on the activity of cuttlefish optic lobe neurones is discussed in the context of comparable data from vertebrate preparations and the actions of other neuromodulators in the cuttlefish brain. PMID- 15755518 TI - EEG and postural correlates of mild traumatic brain injury in athletes. AB - Mild traumatic brain injury (MTBI), or concussion, is one of the least understood injuries facing the neuroscience and sports medicine community today. The notion of transient dysfunction and rapid symptom resolution is misleading since symptom resolution is not indicative of injury resolution. Our working hypothesis is that there are residual postural and EEG abnormalities in concussed individuals that could be reliably assessed using appropriate research methodology. This paper presents combined postural and electroencephalographic (EEG) findings suggesting the persistent functional deficits in athletes suffering from MTBI. Twelve concussed athletes and twelve normal controls participated in the study. There was a decrease in EEG power in all bandwidths studied in concussed subjects, especially in standing postures. This was accompanied by sustained postural instability especially under the no vision testing condition. Overall, this study demonstrated the presence of long-term functional abnormalities in individuals suffering from mild traumatic brain injury. PMID- 15755519 TI - Adrenalectomy-induced ZnT3 downregulation in mouse hippocampus is followed by vesicular zinc depletion. AB - The effects of adrenalectomy (ADX) on the vesicular zinc content of zinc-enriched (ZEN) terminals in mouse hippocampus were investigated at light microscopic levels using zinc transporter-3 immunohistochemistry (ZnT3IHC) and zinc selenium autometallography (ZnSeAMG). ZnT3 resides in the synaptic vesicle membranes of ZEN neurons and is believed to move zinc ions into the vesicles. ZnT3IHC staining closely corresponds to the ZnSeAMG staining, but in the present study we present evidence of a delayed decrease of ZEN zinc, as compared to downregulation of the ZnT3 protein following ADX. Twenty-four hours after adrenalectomy the level of ZnT3IHC was visibly reduced while the ZnSeAMG staining intensity seemed unchanged. After 10 and 30 days, however, downregulation of ZnT3 was paralleled by a distinct reduction in ZnSeAMG staining. The total protein concentration of ZnT3 was reduced by about 53%, and the total zinc concentration in the hippocampus of the same mice was reduced by 43-64%, 30 days after the adrenalectomy. The present results support previous results suggesting that ZnT3 is responsible for transport of zinc ions into a pool of synaptic vesicles in ZEN terminals. PMID- 15755520 TI - Expression of ciliary neurotrophic factor (CNTF), CNTF receptor alpha (CNTFR alpha) following experimental intracerebral hemorrhage in rats. AB - Ciliary neurotrophic factor (CNTF) is known as a neuro-survival factor in the developing and developed CNS, as well as in the CNS following injury. However, little is known about the expression of CNTF or that of its receptor (CNTFR alpha) in cases of intracerebral hemorrhage (ICH). We investigated the temporal and spatial profiles of CNTF and CNTFR-alpha expression using a collagenase induced ICH rat model. CNTF expression was up-regulated from the day following ICH induction and reached a peak level at 5 to 14 days, with increased expression observed in brain tissue surrounding the hematoma lesion and white matter structures in association with astroglial proliferation. Further, CNTFR-alpha was transiently expressed in the cerebral cortex surrounding the hematoma, with a peak at 5 days. Administration of exogenous CNTF into the lesion following initiation of ICH resulted in a prolonged expression of CNTFR-alpha on cortical neurons neighboring the hematoma. Our findings suggest differential regulation of CNTF and CNTFR-alpha, and the possibility of a therapeutic strategy using CNTF administration for ICH. PMID- 15755521 TI - Glutamatergic chloride currents associated to glutamate transport? AB - Especially in arthropod glutamatergic synaptic systems, microM l-glutamate (Glu) concentrations often elicit Cl- currents, in addition to the excitatory cationic currents that are triggered by much higher Glu concentrations. In crayfish, Ibotenate (Ibo) is a specific agonist of the Glu-ergic Cl- currents. Application of Glu to Glu-transporters opens associated Cl- currents that inhibit quantal release presynaptically and by occupying the transporter prevents removal of released Glu. The latter prolongs the decay of postsynaptic EPSCs. It was tested whether the Ibo-elicited Cl- currents show the same pre- and post-synaptic effects as the transporter elicited ones, suggesting that also this current component arises through transporter activation. Indeed, Ibo applied to single synaptic junctions produced inhibition of quantal release and prolongation of EPSCs, very similar to the effects of Glu. It seems probable, therefore, that at least in crayfish Glu-ergic Cl- currents are generated by activation of transporters. Since generally such transporters are located around Glu-ergic synapses, this is likely to be a general mechanism. The toxin Ivermectin also elicits Cl- currents. However, while Ivermectin inhibits release too, it does not prolong the decay of EPSCs and is probable to activate GABAergic channels. PMID- 15755523 TI - Involvement of ionotropic glutamate receptors in low frequency electroacupuncture analgesia in rats. AB - The present study was conducted to determine whether blockage of both N-methyl-D aspartate (NMDA) and alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid/kainate (AMPA/KA) receptors influences the induction of low frequency electroacupuncture (EA) analgesia. Although neither intrathecal injection of NMDA antagonist D-2-amino-5-phosphonopentanoic acid (D-AP-5) or AMPA/KA antagonist 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonami-de (NBQX) disodium alone had an effect on analgesia, spinal application of D-AP-5 and NBQX disodium significantly prevented analgesia induced by 2 Hz EA. The intrathecal injection of the excitatory amino acid NMDA produced analgesia for several minutes after intrathecal injection, as did EA stimulation. These results suggest that ionotropic glutamate receptors may be involved in the induction of 2 Hz EA analgesia. PMID- 15755522 TI - The effect of acupuncture on anxiety and neuropeptide Y expression in the basolateral amygdala of maternally separated rats. AB - Recent studies have suggested that maternally deprived rats exhibit anxiogenic like behavior when exposed to stress in later life. Neuropeptide Y (NPY) is involved in the regulation of various physiological functions such as the expression of anxiety. Female Wistar rat pups were separated from their mothers for 3h daily from postnatal days 3 (P3) to 14 (P14). Acupuncture groups were treated with acupuncture at Shenmen (HT7) or Zusanli (ST36) on alternate days from P50 to P62. Their anxiety-like behavior was evaluated using an elevated plus maze at P62, and then NPY immunohistochemistry in the basolateral amygdala (BLA) was performed. Rats exposed to maternal separation (MS) were less likely to explore the open arms of the plus-maze compared to control rats that were not exposed to MS. Among maternally separated groups, the percentage of time spent in the open arms was significantly increased in the HT7 acupuncture group, but not the ST36 acupuncture group, compared to MS group. In accordance with this behavior, the numbers of NPY-immunoreactive cells in the BLA were lower in the MS group compared to the control group. Among maternally separated groups, the numbers of NPY-immunoreactive cells in the BLA were significantly higher in the HT7 acupuncture group, but not higher in the ST36 acupuncture group, compared to MS group. These findings suggest that acupuncture treatment might reduce anxiety like behavior in adult rats following maternal separation by modulating the NPY system in the amygdala. PMID- 15755524 TI - Transient expression of MIDC-8 in the normal mouse brain. AB - In this study, we have immunohistochemically characterized the expression of mononuclear phagocyte markers CD14, CD36, CD68, CD204 and MARCO by parenchymal microglia in the developing and adult mouse brain. We further investigated whether these cells express two well-characterized phenotypic markers of dendritic cells: CD205 (DEC-205/NLDC-145) and MIDC-8 antigen. Our results confirm the lack of expression of dendritic cell markers by microglia. We noted that these cells do not appear to express markers associated with monocytes and macrophages during the course of development, but do express CD68 and CD204 antigens in the adult. Unexpectedly, we also noted the transient expression of MIDC-8 antigen on cells within the medial ganglionic eminence and by neuroepithelial cells lining the lateral ventricles and in the medial lemniscus between E15 and E19. We discuss this finding in the context of neural and haematopoietic differentiation. PMID- 15755525 TI - Sub-chronic nicotine-induced changes in regional cerebral blood volume and transversal relaxation time patterns in the rat: a magnetic resonance study. AB - This preliminary study describes magnetic resonance imaging (MRI) data on the effects of sub-chronic nicotine administration in rats. Nicotine 0.4 mg/kg s.c. free base was given once a day for 4 days to Wistar adult male rats. On day 5, anaesthetized subjects were observed using an MRI tomography system. Regional cerebral blood volume (rCBV) and transversal relaxation time (T2) MRI parameters were measured. Nicotine treatment increased T2 values, with a significant effect in the cingulate cortex. A trend to increase was also observed in the prefrontal cortex and nucleus accumbens. Similarly, the effect of nicotine on rCBV was a significant increase in values compared to saline treatment. Post hoc analysis showed a significant effect of nicotine in the prefrontal cortex, cingulate cortex, mediodorsal thalamus and lateral posterior thalamus. This study showed for the first time that sub-chronic nicotine administration can induce changes in MRI pattern which are (i) similar to human MRI studies, and (ii) common to those described for markers of neuronal metabolic activation in corticolimbic brain regions known to be involved in nicotine dependence. PMID- 15755526 TI - Gq alpha subunit mediates receptor site-specific adaptation in the sugar taste receptor cell of the blowfly, Phormia regina. AB - The gustatory system is essential for almost all animals. The recent identification of G protein-coupled receptor proteins (GPCRs) has progressed molecular biological studies of gustatory systems, although the signal transduction mechanisms have not yet been fully elucidated. In vision and olfactory receptor cells, Gq class G protein is known to be a major signal transducer. By functional blocking of intrinsic Gq with an anti-Gq/11alpha antibody, we investigated the roles of Gq in the sugar receptor cell of the blowfly, Phormia regina. Before and after introduction of the anti-Gq/11alpha antibody into the cell through the DOC-permeabilized cell membrane, we recorded the responses of the receptor cell to sucrose and d-fructose, which stimulate different receptor sites, respectively. The initial impulse frequency in response to either sucrose or D-fructose was not changed by antibody introduction, whereas the adaptation rate in sucrose stimulation, but not fructose stimulation, became slower after antibody introduction. These results indicate that: (1) Gq is a regulator of adaptation in the sugar receptor cell of Phormia, rather than a transducer, and (2) different adaptation mechanisms are promoted by stimulations with sucrose and D-fructose. PMID- 15755527 TI - Ischemic preconditioning as induction of ischemic tolerance after transient ischemic attacks in human brain: its clinical relevance. AB - Ischemic preconditioning has been well-established in different animal models of cerebral ischemia. Suggesting that transient ischemic attack (TIAs) may represent a clinical model of ischemic tolerance, patients with ipsilateral TIA before ischemic stroke may have a better neurological outcome compared to patients without TIA before ischemic stroke. A retrospective case-control study. Analysis of clinical and neuroradiological findings of 130 patients treated with local intra-arterial thrombolysis using urokinase after ischemic stroke between January 2000 and May 2002. Eleven patients (8%) demonstrated prestroke ipsilateral TIA in the same vascular territory. Despite no significant differences in baseline characteristics, independence (modified Ranking Scale score) and neurological outcome (NIHS score) were significantly associated with prior TIA in multivariante models. Patients with prestroke TIA, showing a similar clinical picture (NIHS score) on admission, demonstrated a more favorable outcome (NIHS score (p<0.001) and modified Ranking score (p<0.0001)) compared to patients with unheralded ischemic stroke. The interval between TIA and ischemic stroke (1-7 days), duration of TIAs (0-20 min) and number of TIAs (2-3) influenced the better outcome in this patients' subgroup. In addition, we could show an induction of ischemic tolerance by prestroke ASA treatment in patients without previous new onset TIA. Two to three ipsilateral TIA of the same vascular territory and of up to 20 min duration within 1-7 days may lead to ischemic tolerance to a subsequent ischemic stroke as demonstrated by significantly diminished infarct extention throughout all cortical imagines in neuroradiological studies. The role of ASA has to been further elucidated. PMID- 15755529 TI - Utilizing data grid architecture for the backup and recovery of clinical image data. AB - Grid Computing represents the latest and most exciting technology to evolve from the familiar realm of parallel, peer-to-peer and client-server models. However, there has been limited investigation into the impact of this emerging technology in medical imaging and informatics. In particular, PACS technology, an established clinical image repository system, while having matured significantly during the past ten years, still remains weak in the area of clinical image data backup. Current solutions are expensive or time consuming and the technology is far from foolproof. Many large-scale PACS archive systems still encounter downtime for hours or days, which has the critical effect of crippling daily clinical operations. In this paper, a review of current backup solutions will be presented along with a brief introduction to grid technology. Finally, research and development utilizing the grid architecture for the recovery of clinical image data, in particular, PACS image data, will be presented. The focus of this paper is centered on applying a grid computing architecture to a DICOM environment since DICOM has become the standard for clinical image data and PACS utilizes this standard. A federation of PACS can be created allowing a failed PACS archive to recover its image data from others in the federation in a seamless fashion. The design reflects the five-layer architecture of grid computing: Fabric, Resource, Connectivity, Collective, and Application Layers. The testbed Data Grid is composed of one research laboratory and two clinical sites. The Globus 3.0 Toolkit (Co-developed by the Argonne National Laboratory and Information Sciences Institute, USC) for developing the core and user level middleware is utilized to achieve grid connectivity. The successful implementation and evaluation of utilizing data grid architecture for clinical PACS data backup and recovery will provide an understanding of the methodology for using Data Grid in clinical image data backup for PACS, as well as establishment of benchmarks for performance from future grid technology improvements. In addition, the testbed can serve as a road map for expanded research into large enterprise and federation level data grids to guarantee CA (Continuous Availability, 99.999% up time) in a variety of medical data archiving, retrieval, and distribution scenarios. PMID- 15755530 TI - International Internet-2 performance and automatic tuning protocol for medical imaging applications. AB - Internet-2 is an advanced computer network, which has been widely used for medical imaging applications such as teleradiology and teleconsultation, since Internet-2 can fulfill the requirements for high-speed data transmission and short turn-around time with low operation cost once installed. However, such high performance of Internet-2 may not be retained for global access from international network peers. Considering the international Internet-2 connection between the PolyU and the IPI/USC, there exist two major factors, network looping in the US and bottleneck of the connection, raising the round-trip time and limiting the available bandwidth, respectively. The available bandwidth will be further underutilized if the TCP/IP parameters at the sending and receiving computers are not appropriately chosen. This paper proposes a repeatable and consistent protocol to automatically tune these parameters for the clinical applications. PMID- 15755531 TI - Web-based electronic patient records for collaborative medical applications. AB - We developed a web-based system to interactively display electronic patient records (EPR), such as DICOM images, graphics, and structure reports and therapy records, for intranet and internet collaborative medical applications. This system has three major components, a C/S (client/server) architecture for EPR data acquisition and authoring, and a Web B/S architecture for data delivering. The Web viewer of this system integrates multi-media display modules and remote control module together to provide interactive EPR display and manipulation functions for collaborative applications. We have successfully used this system two times to provide teleconsultation for severe acute respiratory syndrome (SARS) patients in Shanghai Infection Hospital and Xinhua Hospital. During the consultation, both the physicians in infection control area and the experts outside the control area could use this system interactively to manipulate and navigate the EPR objects of the SARS patients to facilitate a more precise diagnosis. This paper gives a new approach to create and manage image-based EPR from actual patient records, and also presents a novel method to use Web technology and DICOM standard to build an open architecture for collaborative medical applications. The system can be used for both intranet and internet medical applications such as tele-diagnosis, teleconsultation, and distant learning. PMID- 15755532 TI - A model of DICOM-based electronic patient record in radiation therapy. AB - Electronic patient records for radiation therapy (RT) consists of text, images and graphics. To enable the exchange of RT patient information between systems and institutions, a common standard is called for and the DICOM standard extension to radiation therapy is an appropriate means for standardization. This paper describes a model of DICOM-based electronic patient record system for information exchange and sharing. The system used a DICOM-based RT archive server as a common platform for archival of all RT related information including images and the web technology for distribution and viewing of the patient electronic record. PMID- 15755533 TI - Integrating images into the electronic patient record of the hospital authority of Hong Kong. AB - Over a period of 10 years the Hospital Authority has developed an integrated clinical information system (The Clinical Management System-CMS), including a territory-wide longitudinal electronic patient record (ePR). The CMS and ePR are used by over 4000 doctors and 20,000 other clinicians to document and review care. The ePR currently has nearly 3TB of data covering 44 million episodes for 6.4 million patients. To date the CMS has largely dealt with textual data. However, PACS technology is increasingly being adopted in the HA. This paper describes our strategy for taking the hospital image archives and making them available throughout the Authority as part of the longitudinal patient record, leveraging the ePR to distribute radiological and other images in a manner which is integrated, affordable and sustainable. PMID- 15755534 TI - Automatic categorization of medical images for content-based retrieval and data mining. AB - Categorization of medical images means selecting the appropriate class for a given image out of a set of pre-defined categories. This is an important step for data mining and content-based image retrieval (CBIR). So far, published approaches are capable to distinguish up to 10 categories. In this paper, we evaluate automatic categorization into more than 80 categories describing the imaging modality and direction as well as the body part and biological system examined. Based on 6231 reference images from hospital routine, 85.5% correctness is obtained combining global texture features with scaled images. With a frequency of 97.7%, the correct class is within the best ten matches, which is sufficient for medical CBIR applications. PMID- 15755535 TI - Informatics infrastructure of CAD system. AB - A computer aided diagnosis (CAD) system requires several components which influence its effectiveness. An image processing methodology is responsible for the analysis, database structure archives and distributes the patient demographics, clinical information, and image data. A graphical user interface is applied in order to enter the data and present it to the user. By designing dynamic Web pages a remote access to the entire is granted. The computer aided diagnosis system includes three layers, which might be installed on various platforms. Elements of the application software are designed independently. Integration of all components is another issue discussed in the presented paper. Implementation of a computer aided diagnosis system improves and accelerates the analysis by giving to the user objective measurement tools. It also standardizes the decision-making process and solves the problem of replicability. Finally, it permits a set of images and features to be collected and recognized as a medical standard and be applied in education and research. PMID- 15755536 TI - Image informatics at a national research center. AB - Image informatics at the Communications Engineering Branch of the Lister Hill National Center for Biomedical Communications (LHNCBC), an R&D division of the National Library of Medicine (NLM), includes document and biomedical images. In both domains, research into computer-assisted methods for information extraction, and the implementation of prototype systems incorporating such methods, is central to our mission. Current document image research focuses on extracting bibliographic data from scanned journal articles. Current biomedical imaging work focuses on content-based image retrieval (CBIR) and related problems in segmentation, indexing, and classifying collections of images of the spine and of the uterine cervix. PMID- 15755537 TI - Image-matching as a medical diagnostic support tool (DST) for brain diseases in children. AB - Imaging-matching is an important research area in imaging informatics. We have developed and evaluated a novel diagnostic support tool (DST) based on medical image matching using MR brain images. The approach consists of two steps, database generation and image matching. The database contains pre-diagnosed MR brain images. As the images are added to the database, they are registered to the 3D Talairach coordinate system. In addition, regions of interests (ROI) are generated, and image-processing techniques are used to extract relevant image parameters related to the brain and diseases from the ROIs and from the entire MR image. The second step is to retrieve relevant information from the database by performing image matching. In this step, the physician first submits a query image. The DST computes the similarity between the query image and each of the images in the database, and then presents the most similar images to the user. Since the database contains pre-diagnosed images, the retrieved cases tend to contain relevant diagnostic information. To evaluate the usefulness of the DST in a clinical setting, pediatric brain diseases were used. The database contains 2500 pediatric patients between ages 0 and 18 with brain Magnetic Resonance (MR) images of known brain lesions. A testbed was established at the Children's Hospital Los Angeles (CHLA) for acquiring MR images from the PACS server of patients with known lesions. These images were matched against those in the DST pediatric brain MR database. An expert pediatric neuroradiologist evaluated the matched results. We found that in most cases, the image-matching method was able to quickly retrieve images with relevant diagnostic content. The evaluation method and results are given. PMID- 15755538 TI - User interface paradigms for patient-specific surgical planning: lessons learned over a decade of research. AB - This paper covers work in virtual reality-based, patient-specific surgical planning over the past decade. It aims to comprehensively examine the user interface paradigms and system designs during that period of time and to objectively analyze their effectiveness for the task. The goal is to provide useful feedback on these interface and implementation paradigms to aid other researchers in this field. First, specialized systems for specific clinical use were produced with a limited set of visualization tools. Later, through collaboration with NASA, an immersive virtual environment was created to produce high-fidelity images for surgical simulation, but it underestimated the importance of collaboration. The next system, a networked, distributed virtual environment, provided immersion and collaboration, but the immersive paradigm was found to be of a disadvantage and the uniqueness of the framework unwieldy. A virtual model, workbench-style display was then created using a commercial package, but limitations of each were soon apparent. Finally, a specialized display, with an integrated visualization and simulation system is described and evaluated. Lessons learned include: surgical planning is an abstract process unlike surgical simulation; collaboration is important, as is stereo visualization; and that high-resolution preoperative images from standard viewpoints are desirable, but interaction is truly the key to planning. PMID- 15755539 TI - Designing high-quality interactive multimedia learning modules. AB - Modern research has broadened scientific knowledge and revealed the interdisciplinary nature of the sciences. For today's students, this advance translates to learning a more diverse range of concepts, usually in less time, and without supporting resources. Students can benefit from technology-enhanced learning supplements that unify concepts and are delivered on-demand over the Internet. Such supplements, like imaging informatics databases, serve as innovative references for biomedical information, but could improve their interaction interfaces to support learning. With information from these digital datasets, multimedia learning tools can be designed to transform learning into an active process where students can visualize relationships over time, interact with dynamic content, and immediately test their knowledge. This approach bridges knowledge gaps, fosters conceptual understanding, and builds problem-solving and critical thinking skills-all essential components to informatics training for science and medicine. Additional benefits include cost-free access and ease of dissemination over the Internet or CD-ROM. However, current methods for the design of multimedia learning modules are not standardized and lack strong instructional design. Pressure from administrators at the top and students from the bottom are pushing faculty to use modern technology to address the learning needs and expectations of contemporary students. Yet, faculty lack adequate support and training to adopt this new approach. So how can faculty learn to create educational multimedia materials for their students? This paper provides guidelines on best practices in educational multimedia design, derived from the Virtual Labs Project at Stanford University. The development of a multimedia module consists of five phases: (1) understand the learning problem and the users needs; (2) design the content to harness the enabling technologies; (3) build multimedia materials with web style standards and human factors principles; (4) user testing; (5) evaluate and improve design. PMID- 15755540 TI - HIPAA compliant auditing system for medical images. AB - As an official regulation for healthcare privacy and security, Health Insurance Portability and Accountability Act (HIPAA) mandates health institutions to protect health information against unauthorized use or disclosure. One such method proposed by HIPAA Security Standards is audit trail, which records and examines health information access activities. HIPAA mandates healthcare providers to have the ability to generate audit trails on data access activities for any specific patient. Although current medical imaging systems generate activity logs, there is a lack of formal methodology to interpret these large volumes of log data and generate HIPAA compliant auditing trails. This paper outlines the design of a HIPAA compliant auditing system (HCAS) for medical images in imaging systems such as PACS and discusses the development of a security monitoring (SM) toolkit based on some of the partial components in HCAS. PMID- 15755541 TI - A "deficient environment" in prenatal life may compromise systems important for cognitive function by affecting BDNF in the hippocampus. AB - The intrauterine environment has the capacity to mold the prenatal nervous system. Particularly, recent findings show that an adverse prenatal environment produces structural defects of the hippocampus, a critical area sub-serving learning and memory functions. These structural changes are accompanied by a disruption in the normal expression pattern of brain-derived neurotrophic factor (BDNF) and its cognate tyrosine kinase B (TrkB) receptor. The important role that the BDNF system plays in neural modeling and learning and memory processes suggests that fetal exposure to unfavorable intrauterine conditions may compromise proper cognitive function in adult life. These findings have implications for disorders that involve a dysfunction in the BDNF system and are accompanied by cognitive deficits. PMID- 15755542 TI - alpha-Synuclein: a potent inducer of tau pathology. PMID- 15755543 TI - Neurogenesis in the ependymal layer of the adult rat 3rd ventricle. AB - Neurogenesis has been described in limited regions of the adult mammalian brain. In this study, we showed that the ependymal layer of the 3rd ventricle is a neurogenic region in the adult rat brain. DiI labeling of the 3rd ventricle revealed that neural progenitor cells were derived from cells at the ependymal layer of the adult 3rd ventricle. The mitosis of these progenitor cells at the ependymal layer was promoted by bFGF administration. Combination of BrdU administration, nestin/GFAP immunohistochemistry, and labeling by GFP-recombinant adenoviral infection (vGFP) indicated that at least some tanycytes might be neural progenitor cells in the ependymal layer of the 3rd ventricle. Tracing by vGFP indicated that neural progenitor cells may have migrated from the 3rd ventricle to the hypothalamic parenchyma, where they were integrated into neural networks by forming synapses. In addition, some BrdU(+) neurons had immunoreactivity for orexin A in the hypothalamus. These results indicate that neural progenitor cells exist in the ependymal layer of the adult rat 3rd ventricle and that they may differentiate into neurons functioning in the hypothalamus. PMID- 15755544 TI - BDNF and TrkB protein expression is altered in the fetal hippocampus but not cerebellum after chronic prenatal compromise. AB - This study examines the effects of a chronic prenatal insult on both the expression of brain-derived neurotrophic factor (BDNF) and TrkB proteins and the structural development of the fetal hippocampus and cerebellum. Chronic placental insufficiency (CPI) was induced via unilateral ligation of the uterine artery from midgestation to near term in the pregnant guinea pig. Fetuses were delivered at 60 days of gestation (dg, term approximately 67 dg) and classified as control or growth-restricted (GR) according to established criteria. In hippocampi and cerebella from control (n = 7) and GR (n = 8) fetuses, immunohistochemistry was performed to detect the expression of BDNF and TrkB proteins, and the growth of neuropil and cellular layers was measured in each structure. The growth of neuropil layers was reduced in the dentate gyrus of GR fetuses compared to controls: hippocampi from severely GR fetuses showed a marked reduction in BDNF IR and an increase in TrkB-IR. The most pronounced effects on neuropil growth were seen in the same fetuses that demonstrated marked alterations in BDNF-IR and TrkB-IR. In the cerebellum, there were significant reductions in the growth of the cellular and neuropil layers; however, BDNF-IR and TrkB-IR were not affected. These results demonstrate that CPI has a widespread effect in retarding process growth in the developing brain, but a differential effect on neurotrophin expression. Changes in BDNF and TrkB expression appear to be associated with the pronounced structural changes in the hippocampi of severely GR fetuses, however, structural abnormalities in the cerebellum were not associated with changes in these proteins; presumably, other factors are involved. PMID- 15755545 TI - Tau phosphorylation increases in symptomatic mice overexpressing A30P alpha synuclein. AB - Mice overexpressing mutant alpha-synuclein develop a progressive loss of motor function associated with the accumulation of aggregated alpha-synuclein in neurons of the brainstem. Recent reports suggest that tau pathology might also be associated with Parkinson disease (PD) and aggregation of alpha-synuclein. We now report that mice overexpressing A30P alpha-synuclein develop abnormally phosphorylated tau in parallel with the accumulation of aggregated alpha synuclein. Enhanced phosphorylation of tau occurs only in symptomatic mice that also harbor abundant aggregated alpha-synuclein. The increased phosphorylation of tau occurs at S396/404 and S202 as shown by immunoblotting and immunocytochemical studies with the antibodies PHF-1 and AT8. Neurons that accumulated alpha synuclein occurred in the dorsal brainstem and did not show strong colocalization with neurons that showed abnormal tau phosphorylation, which largely occurred in the ventral brainstem. Aggregation of alpha-synuclein and phosphorylation of tau are associated with increased levels of phosphorylated c-jun kinase (JNK), which is a stress kinase known to phosphorylate tau protein. These results suggest that alpha-synuclein pathology can stimulate early pathological changes in tau. PMID- 15755546 TI - Optical imaging of epileptiform activity in experimentally induced cortical malformations. AB - Electrophysiological studies of human cortical dysplasia and rodent models revealed widespread hyperexcitability in the malformation itself as well as in its vicinity. We here analyzed the initiation of paroxysmal epileptiform activity using optical imaging of neuronal activity in rats with cortical malformations induced by neonatal freeze lesions. Brain slice preparations were incubated with the voltage-sensitive dye RH795 and neuronal activity was monitored using a fast imaging photodiode array combined with standard field potential recordings. Spontaneous paroxysmal epileptiform activity emerged in all slices from animals with cortical malformations and sham-operated controls 20-40 min after omission of extracellular Mg(2+). Following electrophysiological and optical recordings, slices were histochemically processed. Using this approach, the present study demonstrated that in animals with freeze-lesion-induced focal cortical malformations, paroxysmal epileptiform activity always emerged from the dysplastic cortex and then spread to adjacent areas through superficial layers. This distribution of initiation sites was significantly different to sham operated controls in which epileptogenic foci were located in various cytoarchitectonic areas. The present study indicates that following global changes in excitability, the dysplastic cortex itself is the main initiation site of paroxysmal epileptiform activity in animals with focal cortical malformations. PMID- 15755547 TI - Differential responses of spinal axons to transection: influence of the NG2 proteoglycan. AB - Spinal cord transections were performed in wild type and NG2 proteoglycan null mice in order to study penetration of regenerating axons into the scar that forms in response to this type of injury. Aside from the presence or absence of NG2, the features of the transection scar did not differ between the two genotypes. In both cases, the rostral and caudal spinal cord stumps were separated by collagenous connective tissue that was continuous with the spinal cord meninges. In wild type mice, oligodendrocyte progenitors, macrophages, and microvascular pericytes contributed to up-regulation of NG2 expression in and around the scar. Substantial amounts of non-cell associated NG2 were also observed in the scar. The abilities of two classes of spinal axons to penetrate the transection scar were examined. Serotonergic efferents and calcitonin gene-related peptide positive sensory afferents both were observed within the lesion, with calcitonin gene-related peptide-positive axons exhibiting a greater capability to penetrate deeply into the scar tissue. These observations demonstrate inherent differences in the abilities of distinct types of neurons to penetrate the scar. Significantly, growth of serotonergic axons into the transection scar was observed twice as frequently in wild type mice as in NG2 knockout mice, suggesting a stimulatory role for the proteoglycan in regeneration of these fibers. These findings run counter to in vitro evidence implicating NG2 as an inhibitor of nerve regeneration. This work therefore emphasizes the importance of including in vivo models in evaluating the responses of specific types of neurons to spinal cord injury. PMID- 15755548 TI - Ca(2+)/calmodulin-protein kinase IIalpha in the trigeminal subnucleus caudalis contributes to neuropathic pain following inferior alveolar nerve transection. AB - Calcium-calmodulin protein kinase IIalpha (CaMKIIalpha) is mainly found in brain cells, and the mRNA concentrates highly in the postsynaptic density. CaMKIIalpha is an effector of calcium and calmodulin mediated functions, and the phosphorylated CaMKIIalpha (pCaMKIIalpha) activates glutamate receptors, such as the AMPA receptor, and enhances its function. In the present study, we examined whether CaMKIIalpha in trigeminal brainstem neurons contributed to the neuropathic pain induced by inferior alveolar nerve (IAN) transection. Using immunohistochemistry and in situ hybridization, we found that the expression of CaMKIIalpha and pCaMKIIalpha increased in the trigeminal subnucleus caudalis (Vc) after IAN transection. The significant increase in the protein of CaMKIIalpha peaked at 30 min after IAN transection, and the mRNA of CaMKIIalpha increased from 2 to 14 days. Double immunofluorescent staining for CaMKIIalpha and MAP2, a marker of dendrite, revealed a significant increase in the overlapping area at 30 min after injury. This suggests that CaMKIIalpha protein is synthesized from the local mRNA pool in the dendrite 30 min after IAN transection and may quickly transmit information after nerve injury. In the behavioral test in which the escape threshold from mechanical stimulation to the lateral face was measured, intrathecal administration of KN-93, a CaMKII inhibitor, for 7 days significantly inhibited mechano-allodynia induced by IAN transection, as compared with administration of a control peptide. These data suggest that CaMKIIalpha in the trigeminal subnucleus caudalis may be involved in neuropathic pain caused by IAN transection. PMID- 15755549 TI - Genetics of subthalamic nucleus in development and disease. AB - The subthalamic nucleus (STN) is a crucial node in the basal ganglia. Clinical success in targeting the STN for deep brain stimulation in Parkinson's disease patients has prompted increased interest in understanding STN biology. In this report, we discuss recent evidence for transcription factor mediated regulation of STN development. We also review STN developmental neurobiology and known patterns of gene expression in the developing and mature STN. PMID- 15755550 TI - Minocycline delays disease onset and mortality in reovirus encephalitis. AB - Minocycline is neuroprotective in many experimental models of neurodegenerative diseases and central nervous system (CNS) injury but has not previously been tested in a model of viral encephalitis. Experimental infection of neonatal mice with neurotropic reoviruses is a classic model for studying the pathogenesis of viral encephalitis. Intracerebral inoculation of serotype 3 reovirus strain Dearing (T3D) in neonatal mice results in lethal encephalitis caused by neuronal apoptosis throughout the CNS. Minocycline significantly delayed death in mice to 11.6 +/- 0.9 days post-infection vs. 8.6 +/- 0.7 days post-infection in controls (P < 0.01). Virus-induced CNS injury, apoptosis, viral titer and antigen expression were significantly decreased in the brains of minocycline-treated mice on 6 and 8 days post-infection compared to controls. Virus-induced injury and viral titer in minocycline-treated infected mice at 11 days post-infection were similar to those seen in untreated T3D-infected mice at 8 days post-infection. Little microglial or astrocytic invasion of brain regions with viral injury was found at any time-point in untreated or minocycline-treated mice, suggesting that in this model system the neuroprotective effect exerted by minocycline is more likely due to its anti-apoptotic properties rather than its capacity to inhibit microglial activation and limit gliosis. These findings, similar to those reported for neurodegenerative diseases, indicate that minocycline does not prevent development of fatal reovirus encephalitis but delays disease onset and progression, suggesting that minocycline treatment may provide a useful adjunctive therapy in viral CNS infections. PMID- 15755551 TI - Upregulation of activating transcription factor 3 (ATF3) by intrinsic CNS neurons regenerating axons into peripheral nerve grafts. AB - The expression of the transcription factor ATF3 in the brain was examined by immunohistochemistry during axonal regeneration induced by the implantation of pieces of peripheral nerve into the thalamus of adult rats. After 3 days, ATF3 immunoreactivity was present in many cells within approximately 500 mum of the graft. In addition, ATF3-positive cell nuclei were found in the thalamic reticular nucleus (TRN) and medial geniculate nuclear complex (MGN), from which most regenerating axons originate. CNS cells with ATF3-positive nuclei were predominantly neurons and did not show signs of apoptosis. The number of ATF3 positive cells had declined by 7 days and further by 1 month after grafting when most ATF3-positive cells were found in the TRN and MGN. 14 days or more after grafting, some ATF3-positive nuclei were distorted and may have been apoptotic. In some experiments of 1 month duration, neurons which had regenerated axons to the distal ends of grafts were retrogradely labeled with DiAsp. ATF3-positive neurons in these animals were located in regions of the TRN and MGN containing retrogradely labeled neurons and the great majority were also labeled with DiAsp. SCG10 and c-Jun were found in neurons in the same regions as retrogradely labeled and ATF3-positive cells. Thus, ATF3 is transiently upregulated by injured CNS neurons, but prolonged expression is part of the pattern of gene expression associated with axonal regeneration. The co-expression of ATF3 with c-jun suggests that interactions between these transcription factors may be important for controlling the program of gene expression necessary for regeneration. PMID- 15755552 TI - Increased neurogenesis and the ectopic granule cells after intrahippocampal BDNF infusion in adult rats. AB - There is evidence that BDNF influences the birth of granule cells in the dentate gyrus, which is one of the few areas of the brain that demonstrates neurogenesis throughout life. However, studies to date have not examined this issue directly. To do so, we compared the effects of BDNF, phosphate-buffered saline (PBS), or bovine serum albumin (BSA) on neurogenesis after infusion into the hippocampus of the normal adult rat, using osmotic pumps that were implanted unilaterally in the dorsal hilus. BDNF, PBS, and BSA were infused for 2 weeks. The mitotic marker bromodeoxyuridine (BrdU) was administered twice daily during the 2-week infusion period. At least 1 month after infusion ended, brains were processed immunocytochemically using antibodies to BrdU, a neuronal nuclear protein (NeuN), or calbindin D28K (CaBP), which labels mature granule cells. Stereology was used to quantify BrdU-labeled cells in the dorsal hippocampus that were double-labeled with NeuN or CaBP. There was a statistically significant increase in BrdU(+)/NeuN(+) double-labeled cells in the granule cell layer after BDNF infusion relative to controls. The values for BrdU(+)/NeuN(+) cells were similar to BrdU(+)/CaBP(+) cells, indicating that most new neurons were likely to be granule cells. In addition, BrdU(+)/NeuN(+)-labeled cells developed in the hilar region after BDNF infusion, which have previously only been identified after severe continuous seizures (status epilepticus) and associated pathological changes. Remarkably, neurogenesis was also increased contralaterally, but BDNF did not appear to spread to the opposite hemisphere. Thus, infusion of BDNF to a local area can have widespread effects on hippocampal neurogenesis. The results demonstrate that BDNF administration to the dentate gyrus leads to increased neurogenesis of granule cells. They also show that ectopic granule cells develop after BDNF infusion, which suggests that ectopic migration is not necessarily confined to pathological conditions. These results are discussed in light of the evidence that BDNF increases neuronal activity in hippocampus. Thus, the mechanisms underlying neurogenesis following BDNF infusion could be due to altered activity as well as direct effects of BDNF itself, and this is relevant to studies of other growth factors because many of them have effects on neuronal excitability that are often not considered. PMID- 15755554 TI - Engraftment of freshly isolated or cultured human umbilical cord blood cells and the effect of cyclosporin A on the outcome. AB - Human umbilical cord blood (HUCB) is a potentially valuable resource for cell therapy. The present study investigated the short-term survival of intrastriatal grafts of either freshly isolated or cultured HUCB cells and the effect of the immunosuppressive agent cyclosporin A (CSA) in host rat brains. The group injected with either freshly isolated or cultured HUCB cells was subdivided into CSA or saline controls. Freshly isolated and cultured HUCB cells displayed surface markers CD33, CD44, CD45, CD51/61 and CD90/Thy-1. The hematopoietic progenitor marker CD34 was expressed only in freshly isolated cells. The majority of injected HUCB cells were localized within a 500-mum radius from the injection site in the striatum; however, a subpopulation migrated along the corpus callosum. There was no significant statistical difference in the cell count between freshly isolated and cultured HUCB cells with or without CSA. Some grafted HUCB cells expressed either a neural or microglial marker. There was weak up-regulation of major histocompatibility complex (MHC) class I antigen in rats either with or without CSA. However, there were considerably fewer positive cells labeled with an MHC class II antigen in CSA groups. These results suggest that neither freshly isolated nor cultured HUCB cells induce acute rejection after intrastriatal transplantation up to 14 days. CSA suppressed up-regulation of MHC class II antigen in the host brain. PMID- 15755553 TI - Cell-permeant calcium buffer induced neuroprotection after cortical devascularization. AB - An excitotoxic cascade resulting in a significant intracellular calcium load is thought to be a primary mechanism leading to neuronal death after ischemia. One way to protect neurons from injury is through the use of cell-permeant calcium buffers. These molecules have been reported to be neuroprotective via their ability to increase the cell's overall Ca(2+) buffering load as well as by attenuating neurotransmitter release. However, their efficacy when given after injury has yet to be determined. We used diffusion-weighted magnetic resonance imaging (DWI), histological, and immunohistochemical methods to determine the neuroprotective efficacy of 2-aminophenol-N, N, O-triacetic acid acetoxymethyl ester (APTRA-AM) after focal cerebral ischemia. Injured animals were given two injections of APTRA-AM at 1 and 12 h after injury. Animals were imaged prior to injury and then at 12, 24, 48 h and 3 and 7 days after injury. After 7 days the animals were euthanized for correlative cresyl violet histology and immunohistochemistry. Injury resulted in a decrease in the apparent diffusion coefficient (ADC) of the injured area within the first 12 h of injury, which returned to normal by 7 days. In contrast, animals injected with APTRA-AM showed no significant change in the ADC at any time point studied. Tissue analysis showed that APTRA-AM significantly reduced the infarct size by 85% and extent of inflammatory cell infiltration by 94%. The results clearly demonstrate significant neuroprotection by APTRA-AM when given after injury. PMID- 15755555 TI - Compartmentalization of TCR repertoire alteration during rejection of an intrabrain xenograft. AB - Xenograft rejections of embryonic pig neural cells implanted into the adult rat striatum occurs within 3-4 weeks, following a dramatic T cell infiltration. Little is known about the cross-talk between the brain and peripheral lymphoid tissues which results in this recruitment and lymphocyte homing. To better characterize the dynamics of the T cell response against xenogeneic neural cells implanted into the brain parenchyma, we used both qualitative and quantitative methods to follow the alterations of the CDR3 length distribution (CDR3-LD) of the TCR (T cell receptor) beta chain in the transplanted striatum and compared this response to that observed in the deep cervical lymph nodes, spleen, and blood. Data showed that the T cell repertoire diversity was highly altered in the recipient brain during xenograft rejection. Comparison of the alterations of the CDR3-LD between several animals revealed a single public alteration in the Vbeta20 family, and many private alterations of the CDR3-LD which differed from one infiltrated brain to another. Alterations of the T cell repertoire were also observed in lymphocytes homed into the deep cervical lymph nodes. However, they differed from the alterations detected in the infiltrated brains. Conversely, no significant alteration of the CDR3-LD was detected in the spleen or in the blood. These data suggest that the deep cervical lymph nodes play an active role in the process of xenograft recognition or/and rejection. However, they also indicate that the fate of T cells homed in the brain and deep cervical lymph nodes differs. PMID- 15755556 TI - Demyelination and Schwann cell responses adjacent to injury epicenter cavities following chronic human spinal cord injury. AB - The natural history of post-traumatic demyelination and myelin repair in the human spinal cord is largely unknown and has remained a matter of speculation. A wealth of experimental studies indicate that mild to moderate contusive injuries to the mammalian spinal cord evolve into a cavity with a preserved rim of white matter in which a population of segmentally demyelinated axons persists. It is believed that such injured axons have abnormal conduction properties. Theoretically, such axons might show improved function if myelin repair occurred. Schwann cells can remyelinate axons affected by multiple sclerosis, but little evidence exists that such repair can occur spontaneously following traumatic human SCI. Therefore, it is important to determine if chronic demyelination is present following human spinal cord injury. There are no previous reports that have conclusively demonstrated demyelination in the human spinal cord following traumatic spinal cord injury using immunohistochemical techniques. Immunohistochemical methods were used to study the distribution of peripheral and central myelin proteins as well as axonal neurofilament at the injury epicenter in 13 postmortem chronically injured human spinal cords 1-22 years following injury. Of these seven could be assessed by our methods. We found that some axonal demyelination can be detected even a decade following human SCI and indirect evidence that invading Schwann cells contributed to restoration of myelin sheaths around some spinal axons. PMID- 15755558 TI - A potent sorbitol dehydrogenase inhibitor exacerbates sympathetic autonomic neuropathy in rats with streptozotocin-induced diabetes. AB - We have developed an animal model of diabetic sympathetic autonomic neuropathy which is characterized by neuroaxonal dystrophy (NAD), an ultrastructurally distinctive axonopathy, in chronic streptozotocin (STZ)-diabetic rats. Diabetes induced alterations in the sorbitol pathway occur in sympathetic ganglia and therapeutic agents which inhibit aldose reductase or sorbitol dehydrogenase improve or exacerbate, respectively, diabetes-induced NAD. The sorbitol dehydrogenase inhibitor SDI-711 (CP-470711, Pfizer) is approximately 50-fold more potent than the structurally related compound SDI-158 (CP 166,572) used in our earlier studies. Treatment with SDI-711 (5 mg/kg/day) for 3 months increased ganglionic sorbitol (26-40 fold) and decreased fructose content (20-75%) in control and diabetic rats compared to untreated animals. SDI-711 treatment of diabetic rats produced a 2.5- and 4-5-fold increase in NAD in the SMG and ileal mesenteric nerves, respectively, in comparison to untreated diabetics. Although SDI-711 treatment of non-diabetic control rat ganglia increased ganglionic sorbitol 40-fold (a value 8-fold higher than untreated diabetics), the frequency of NAD remained at control levels. Levels of ganglionic sorbitol pathway intermediates in STZ-treated rats (a model of type 1 diabetes) and Zucker Diabetic Fatty rats (ZDF, a genetic model of type 2 diabetes) were comparable, although STZ-diabetic rats develop NAD and ZDF-diabetic rats do not. SDI failed to increase diabetes-related ganglionic NGF above levels seen in untreated diabetics. Initiation of Sorbinil treatment for the last 4 months of a 9 month course of diabetes, substantially reversed the frequency of established NAD in the diabetic rat SMG without affecting the metabolic severity of diabetes. These findings indicate that sorbitol pathway-linked metabolic alterations play an important role in the development of NAD, but sorbitol pathway activity, not absolute levels of sorbitol or fructose per se, may be most critical to its pathogenesis. PMID- 15755557 TI - Astrocyte growth effects of vascular endothelial growth factor (VEGF) application to perinatal neocortical explants: receptor mediation and signal transduction pathways. AB - The non-angiogenic role of vascular endothelial growth factor (VEGF), and its receptors flt-1 and flk-1, together with downstream signaling pathways were examined in fetal and postnatal rat cerebral cortical organotypic explants. VEGF application in both paradigms caused a significant increase in astroglial proliferation and a dose-dependent increase in GFAP and nestin immunoreactivity. The VEGF receptor flt-1 was observed on most, though not all astrocytes, while flk-1 receptor immunoexpression was absent. Treatment with antisense oligonucleotides (AS-ODNs) to flt-1 resulted in a dramatic decrease in GFAP and nestin immunoreactivity, which further confirmed the role of flt-1 in mediating VEGF's gliotrophic effects, while AS-ODNs to flk-1 had no effect. VEGF-induced gliotrophic effects were found to be mediated by the MAPK/ERK and PI-3 kinase signaling pathways, since the both the MEK1 inhibitor, PD98059 and the PI-3 kinase inhibitor, Wortmannin abolished VEGF-induced astrocytic GFAP(+) expression. Although high dose VEGF application resulted in strong upregulation of both GFAP and nestin immunoreactivity in astrocytes, overlap of the two proteins was not observed in all cells, suggesting that some of the nestin(+) cells might be neural progenitors. Exposure to VEGF resulted in upregulation of both VEGF and bFGF mRNA at the one-day time point, and bFGF protein by 3 days; VEGF activated astrocytes expressed bFGF to a much greater degree than those in untreated explants. The increased expression of bFGF induced by VEGF, may serve in the proliferation of multipotential neural stem/progenitor cells in vitro. VEGF, an established angiogenic factor, appears to play a significant role in the growth and differentiation of astrocytes in the CNS. PMID- 15755559 TI - Seizures induced in immature rats by homocysteic acid and the associated brain damage are prevented by group II metabotropic glutamate receptor agonist (2R,4R) 4-aminopyrrolidine-2,4-dicarboxylate. AB - The present study has examined the anticonvulsant and neuroprotective effect of group II metabotropic glutamate receptor (mGluR) agonist (2R,4R)-4 aminopyrrolidine-2,4-dicarboxylate (2R,4R-APDC) in the model of seizures induced in immature 12-day-old rats by bilateral intracerebroventricular infusion of dl homocysteic acid (DL-HCA, 600 nmol/side). For biochemical analyses, rat pups were sacrificed during generalized clonic-tonic seizures, approximately 45-50 min after infusion. Comparable time intervals were used for sacrificing the pups which had received 2R,4R-APDC. Low doses of 2R,4R-APDC (0.05 nmol/side) provided a pronounced anticonvulsant effect which was abolished by pretreatment with a selective group II mGluR antagonist LY341495. Generalized clonic-tonic seizures were completely suppressed and cortical energy metabolite changes which normally accompany these seizures were either normalized (decrease of glucose and glycogen) or markedly reduced (an accumulation of lactate). EEG recordings support the marked anticonvulsant effect of 2R,4R-APDC, nevertheless, this was only partial. In spite of the absence of obvious motor phenomena, isolated spikes or even short periods of partial ictal activity could be observed. Isolated spikes could also be seen in some animals after application of 2R,4R-APDC alone, reflecting most likely subclinical proconvulsant activity of this agonist. The neuroprotective effect of 2R,4R-APDC was evaluated after 24 h and 6 days of survival following DL-HCA-induced seizures. Massive neuronal degeneration, as revealed by Fluoro-Jade B staining, was observed in a number of brain regions following infusion of DL-HCA alone (seizure group), whereas 2R,4R-APDC pretreatment provided substantial neuroprotection. The present findings support the possibility that group II mGluRs are a promising target for a novel approach to treating epilepsy. PMID- 15755560 TI - Dexamethasone enhances NT-3 expression in rat hippocampus after traumatic brain injury. AB - The cellular events in traumatic brain injury (TBI) are complicated, and the factors mediating neurotrophins to protect and repair the injured brain cells are only beginning to be identified. This study examined the effect of dexamethasone (DEX) on neurotrophin-3 (NT-3) expression following TBI. Levels of NT-3 mRNA and protein in rat hippocampus were measured using in situ hybridization and immunohistochemistry, respectively. After TBI, the NT-3 mRNA expression was down regulated during the first 24 h. DEX reversed the post-traumatic reduction of NT 3 mRNA expression at 2, 4, 6, and 12 h in the hippocampus, and also decreased the cell death in hippocampal hilum and supraventricular cerebral cortex after 7 days. The NT-3 protein levels generally corresponded to the mRNA levels in the hippocampal region. DEX enhanced the NT-3 expression after TBI, indicating that post-traumatic neuroprotection in the hippocampus is at least partially mediated by NT-3 and thus can be modulated by DEX treatment. PMID- 15755561 TI - Immune and glial cell factors as pain mediators and modulators. AB - A decade ago the attention of pain scientists was focused on a small number of molecules such as prostaglandin and bradykinin as peripheral pain mediators or modulators. These factors were known to be produced by tissue damage or inflammation, and considered responsible for the activation and sensitization of peripheral pain signaling sensory neurons. A small number of molecules were also identified as central pain mediators, most notably glutamate and substance P released from central nociceptive nerve terminals, and, starting at that time, appreciation that nitric oxide might be produced by dorsal horn neurons and act as a diffusible transmitter to increase excitability of central pain circuits. During the last decade evidence has emerged for many novel pain mediators. The old ones have not disappeared, although their roles have been redefined in some cases. Prostaglandin E2 (PGE2), for instance, is now recognized as playing a prominent role in CNS as well as peripheral tissues. The newly identified mediators include a variety of factors produced and released from nonneuronal cells-predominantly immune and glial cells. The evidence is now growing apace that these are important mediators of persistent pain states and can act at a number of loci. Here we review the actions of several of these factors-the pro inflammatory cytokines, some chemokines, and some neurotrophic factors, which, in addition to their traditionally recognized roles, are all capable of changing the response properties of peripheral and central pain signaling neurons. We review these actions, first in periphery, where a substantial literature has accumulated, and then in spinal cord, where the role of factors from nonneuronal cells has only recently been identified as of considerable importance. PMID- 15755564 TI - Rapid progress in new vaccines development, immunisation and immunotherapy. PMID- 15755562 TI - Cobalamin (vitamin B(12)) in subacute combined degeneration and beyond: traditional interpretations and novel theories. AB - Subacute combined degeneration (SCD) is a neuropathy due to cobalamin (Cbl) (vitamin B(12)) deficiency acquired in adult age. Hitherto, the theories advanced to explain the pathogenesis of SCD have postulated a causal relationship between SCD lesions and the impairment of either or both of two Cbl-dependent reactions. We have identified a new experimental model, the totally gastrectomized rat, to reproduce the key morphological features of the disease [spongy vacuolation, intramyelinic and interstitial edema of the white matter of the central nervous system (CNS), and astrogliosis], and found new mechanisms responsible for the pathogenesis of SCD: the neuropathological lesions in TGX rats are not only due to mere vitamin withdrawal but also to the overproduction of the myelinolytic tumor necrosis factor (TNF)-alpha and the reduced synthesis of the two neurotrophic agents, epidermal growth factor (EGF) and interleukin-6. This deregulation of the balance between TNF-alpha and EGF synthesis induced by Cbl deficiency has been verified in the sera of patients with pernicious anemia (but not in those with iron-deficient anemia), and in the cerebrospinal fluid (CSF) of SCD patients. These new functions are not linked to the coenzyme functions of the vitamin, but it is still unknown whether they involve genetic or epigenetic mechanisms. Low Cbl levels have also been repeatedly observed in the sera and/or CSF of patients with Alzheimer's disease or multiple sclerosis, but whether Cbl deficit plays a role in the pathogenesis of these diseases is still unclear. PMID- 15755565 TI - On the need for, and the delivery of, cross-protective vaccines. AB - The rhinoviruses that are instrumental in causing about one-third of the outbreaks of the common cold present us with some 100 or so serotypes whose convalescent sera do not cross-neutralise. A similar situation prevails with the organism that causes gonorrhoea. Both the HIV and the protozoan causing malaria are notorious for their ability to evade the immune system by changes to their antigenic profile. Similarly, we face continual changes in the antigenic determinants of the influenza virus. It is clear that we require vaccine for these diseases that provide protection against a wide variety of basic variants. This can be achieved, as was shown by Arvind Kumar, who, in his PhD project, generated monoclonal antibodies to cross-reacting yet neutralising epitopes of a number of rhinoviruses. Such antibodies also neutralised some Coxsackie viruses as well as some of the types of Poliovirus. This demonstration of feasibility will be explored further in my paper with a view to arriving at a general approach to the production of vaccines whose humoral and cellular responses can neutralise a wide cross-section of serotype variants. PMID- 15755566 TI - DermaVir, a novel HIV immunisation technology. AB - DermaVir is a novel topical immunisation designed to target Langerhans cells (LC), the epidermal precursors of dendritic cells. LC serve as a vehicle to process and transfer antigens from the skin to T cell areas in the lymphoid organs. The HIV DNA delivered by DermaVir to LC expresses most viral regulatory and structural genes and induces T cell-mediated immune responses with broad specificity. In chronically infected macaques, DermaVir administered with antiretroviral drugs, suppressed viral load rebound after treatment interruption and improved survival. DermaVir is a promising antiretroviral treatment approach with a unique mechanism of action for combination with currently used drugs. PMID- 15755567 TI - Antigenome technology: a novel approach for the selection of bacterial vaccine candidate antigens. AB - A novel approach for the identification of protein antigens from bacterial pathogens was previously developed in our laboratory that combines the advantages of full genome coverage and serological antigen identification. We have applied this technology to several bacterial pathogens of the genera Staphylococcus and Streptococcus and have, as a result, defined the "antigenome" of these pathogens. This catalogue defines the most relevant antigenic proteins that are targeted by the human immune system, including their antibody binding sites. The antigenome technology offers an integrated approach for antigen validation in order to select the most promising candidates for the development of subunit vaccines against the targeted bacterial diseases. Using this technology, novel protective antigens were discovered from several important human pathogens. PMID- 15755568 TI - Magnifection--a new platform for expressing recombinant vaccines in plants. AB - Today, plant biotechnology relies on two processes for delivery and expression of heterologous genes in plants: stable genetic transformation and transient infection with viral vectors. Although much faster, the transient route until recently was limited because of virus' low infectivity and its inability to carry average-size or larger transgenes. A recently developed new generation transfection technology overcomes these limitations by relying on Agrobacterium as an infective systemic agent that delivers viral replicons. This improved process is being used to simultaneously start transient gene amplification and high-level expression in all mature leaves of a plant, and such a transfection can be done on an industrial scale. This eclectic technology, called 'magnifection', combines advantages of three biological systems: vector efficiency and efficient systemic DNA delivery of Agrobacterium, speed and expression level/yield of a plant RNA virus, as well as posttranslational capabilities and low production costs of a plant. The proposed process allows for industrial production that does not require genetic modification of plants, that is much faster than previous methods, and that is biologically safe. Numerous applications in the area of vaccine manufacturing are being discussed. PMID- 15755569 TI - Site-specific peptide vaccines for immunotherapy and immunization against chronic diseases, cancer, infectious diseases, and for veterinary applications. AB - United Biomedical, Inc. (UBI) has developed a set of core technologies for the discovery and production of synthetic peptide-based immunotherapeutics and vaccines. These core technologies have led to products that stimulate functional site-directed antibody responses for therapeutic effects. UBI active immunotherapies can be used to modulate physiological processes effective for the control of cell entry by HIV virions, for control of prostate cancer and allergy, and for immunocastration in livestock leading to boar taint elimination and growth promotion in swine. The UBI technologies are also useful to stimulate site directed antibodies against pathogenic agents such as foot-and-mouth disease virus. UBITh Immunotherapeutic peptides were developed as antigens to direct antibody responses against targeted epitopes on self-proteins and viral pathogens that are responsible for biological functions and pathogenicity. A collection of promiscuous UBITh T helper cell epitopes was used to impart these functionally antigenic peptides with immunogenicity. The T cell helper epitopes were covalently linked to the functional antigenic target sites by peptide synthesis, creating well-defined synthetic immunogens. Finally, vaccine formulations were selected appropriate for the delivery of peptide immunogens. Controlled production processes and the means to characterize the final product provide a framework for the GMP-compliant manufacture of UBITh immunotherapeutics and vaccines. PMID- 15755570 TI - Vaccination markers: designing unique antigens to be added to vaccines to differentiate between natural infection and vaccination. AB - Some important vaccines, including ones against tuberculosis (Tb) and foot-and mouth disease (FMD), are not used in many developed countries because there is no simple way to differentiate between vaccine-induced immunity and infection. Eliminating major protein antigens has not succeeded in the cases of Tb, FMD and some other vaccine preparations. Instead of eliminating antigens, novel antigens can be added to vaccines in order to induce a unique immunological response proving that the vaccine has been delivered. Methods for producing such unique antigens are presented and preliminary test results reported. The technique may also be useful for immunologically "branding" animals. PMID- 15755571 TI - Plasmid maintenance systems suitable for GMO-based bacterial vaccines. AB - Live carrier-based bacterial vaccines represent a vaccine strategy that offers exceptional flexibility. Commensal or attenuated strains of pathogenic bacteria can be used as live carriers to present foreign antigens from unrelated pathogens to the immune system, with the aim of eliciting protective immune responses. As for oral immunisation, such an approach obviates the usual loss of antigen integrity observed during gastrointestinal passage and allows the delivery of a sufficient antigen dose to the mucosal immune system. Antibiotic and antibiotic resistance genes have traditionally been used for the maintenance of recombinant plasmid vectors in bacteria used for biotechnological purposes. However, their continued use may appear undesirable in the field of live carrier-based vaccine development. This review focuses on strategies to omit antibiotic resistance determinants in live bacterial vaccines and discusses several balanced lethal plasmid stabilisation systems with respect to maintenance of plasmid inheritance and antigenicity of plasmid-encoded antigen in vivo. PMID- 15755572 TI - Plasmid vaccination of stable HIV-positive subjects on antiviral treatment results in enhanced CD8 T-cell immunity and increased control of viral "blips". AB - Antiviral therapy prolongs suppression of viral replication and allows for significant immune reconstitution but has not been effective in eradicating reservoirs of virus, which produce resurgent viremia when highly active antiretroviral therapy (HAART) is discontinued. Immune-based therapy may provide an additional antiviral effect. We vaccinated stable HIV-positive patients on HAART with an HIV plasmid vaccine to determine safety, immunogenicity, and therapeutic potential. Volunteers received a combination of two HIV DNA plasmid constructs, which drive expression of env/rev and gag/pol genes. The vaccine was well tolerated with no toxicity. CD4 and CD8 lymphocyte counts did not change significantly among volunteers. CD8 MHC class I-restricted responses to HIV antigens were assayed. Eight of 13 vaccinees responded after vaccination with detectable ELISpot result. Importantly, we observed a difference in viral detection events in vaccinated compared to control patients. Three out of the five placebo recipients had "viral blips" (transient elevations of HIV RNA) during follow-up (10/49 assays) while these were only present in one of 13 vaccinees on one occasion (1/130 assays; p<0.04). The decrease in the frequency of transient viremia and failure suggests that DNA immunization with CD8 generating vaccines in HAART-controlled HIV-positive subjects may have therapeutic potential. PMID- 15755574 TI - The US smallpox vaccination program: a review of a large modern era smallpox vaccination implementation program. AB - We describe the US experience with a large-scale smallpox vaccination program in the modern era and quantify the anticipated and unanticipated local and systemic side-effects of smallpox vaccination. In addition, we review unexpected issues, such as the development of myopericarditis discovered during the implementation of this program. These results constitute the largest dataset of a vaccinia vaccination program utilizing calf-lymph derived New York City Board of Health strain vaccine (Dryvax, Wyeth) since the 1970s. These results should inform current and future vaccinia vaccination programs and provide a historical rate of complications against which candidate vaccine side-effects can be compared in future clinical trials. PMID- 15755573 TI - Induction of broadly neutralizing antibodies against measles virus mutants using a polyepitope vaccine strategy. AB - Chimeric molecules expressing multiple copies of the loop-forming hemagglutinin noose epitope (designated as "L"; aa386-400), a protective B cell epitope of the measles virus were generated by recombinant technology. The recombinant polyepitope [L4T4]2 combining two sets of four repeats of the L epitope and with two sets of four repeats of the human promiscuous T cell epitope of tetanus toxoid ("T", tt830-844) was produced in transgenic carrot plants. After intraperitoneal immunization of mice with plant membrane extract, sera neutralized all wild-type viruses. In a modified plaque reduction neutralization assay based on CD150-transfected Vero cells anti-[L4T4]2 sera neutralized all field isolates, irrespective of mutations in the L epitope. Even viruses with a mutation in the contact residues of a neutralizing L-specific monoclonal antibody or two mutations in other positions of the epitope were equally sensitive to neutralization. These results suggest that the multiple copies of the L epitope fold into different conformations that induce a repertoire of B cells diverse enough to overcome the genetic diversity of field viruses. PMID- 15755576 TI - Paediatric infections: prevention of transmission and disease--implications for adults. AB - Congenital and neonatal viral infections usually display their acute manifestations in highly recognisable ways, for example, congenital rubella, cytomegalovirus (CMV), varicella, human immunodeficiency (HIV) and herpes simplex virus (HSV) infection. By contrast, congenital hepatitis B virus (HBV) infection may go undetected for years. Some of these are preventable, but what is not immediately apparent is that the long-term consequences are being prevented as well. The long-term consequences of congenital and neonatal infections include endocrine, immunological and cardiovascular disease, deafness, visual problems, intellectual handicap and cerebral palsy. With the survival of HIV-infected infants into adulthood the long-term consequences will soon be described. Maternally and neonatally transmitted HBV infection predisposes to carriage, liver cirrhosis and hepatocellular carcinoma in young adults. Neonatal HBV vaccination prevents adult cancer. Acquired viral infections may predispose to subsequent lung disease, malabsorption, fertility problems or neurological disability. In the prevention of acquired rubella, varicella, HBV, influenza, poliovirus, measles and hepatitis A, one should mention the added bonus of preventing secondary cases by preventing transmission from infants and children to other children and adults. Preventing paediatric HSV, HBV and HIV infection in females may even be preventing subsequent transmission to future generations. Turning to paediatric bacterial infections, vaccinating infants and young children against pertussis could not only prevent transmission to older children and adults but also break the cycle, which then transmits from adults back to infants and young children. There is evidence that disease in older age groups, including adults, has been prevented by virtue of herd immunity from paediatric vaccination, e.g. Neisseria meningitidis Group C and Streptococcus pneumoniae. The add-on benefits for other generations, including for adults, arising from the prevention of paediatric infections are considerable. PMID- 15755575 TI - Human-derived, plant-produced monoclonal antibody for the treatment of anthrax. AB - The unpredictable nature of bio-terrorism compels us to develop medical countermeasures that will enable authorities to treat individuals exposed to agents such as anthrax. We report the feasibility of producing a protective, human-derived, monoclonal antibody directed against the protective antigen (PA) of Bacillus anthracis in plants. This was achieved by transient expression using agroinfiltration of Nicotiana benthamiana plants. The resulting antibody was able to neutralize toxin activity in vitro and in vivo at a comparable level to that seen for its hybridoma-produced counterpart. PMID- 15755577 TI - A strategic vaccine facility for the UK. AB - This paper describes a proposed Strategic Vaccine Facility (SVF) to provide a capability to the UK to deal with new and emerging disease threats. It would underpin the vaccine manufacturing industry by developing expertise and technology to enable rapid manufacture of small batches of vaccines for emergency use against agents, such as bioterrorist agents and emerging diseases. It would have a rare ability to work with dangerous pathogens under containment, allowing the production of inactivated and live vaccines, which would be difficult in a conventional plant. The facility's output will include vaccine candidates and manufacturing protocols for transfer to industry, small vaccine batches for emergency use or clinical trials, and vaccine reference standards. It would also be available for manufacturing small batches of experimental and public health vaccines for the UK and the developing world, allowing clinical trials to be undertaken against key diseases. PMID- 15755578 TI - Health economic research on vaccinations and immunisation practices--an introductory primer. AB - The economic importance of vaccines lies partly in the burden of disease that can be avoided and partly in the competition for resources between vaccines and other interventions. Up to the 1980s only few economic evaluations had been carried out. Since then the confrontation of most countries with escalating health care costs and tighter budgets have awakened the interest in pharmacoeconomic analysis. Resources used to provide health care are vast but not limitless. When clinicians are asked to participate in decisions for large groups of patients (in a managed care context, in an institution, or at the level of local health authorities), the balance between consumption of resources and the benefits of an intervention is important. Clinicians may use cost-effectiveness and cost-benefit studies to inform such decisions (but not to make them). Because of differences in methods, the presentation of results, and country-specific parameters, economic evaluations of the same vaccination strategy by different groups may have divergent results. Vaccines differ from classical medicines in at least three ways: firstly, there is a longer tradition of economic evaluations for vaccines than for medicines. Some of the most earliest economic studies were carried out in the field of vaccines in the public health arena. Secondly, comparatively fewer central decision makers need to be convinced as compared to drugs. The reason for this being a more centralised process of recommending vaccines and vaccination policies. Thirdly, externalities are more relevant in the field of vaccines. Such externalities may be positive or negative. Positive externalities are present in the case where herd immunity prevents the spread of the disease in the community. We are now undoubtedly in an era of assessment and accountability for all new technologies in healthcare. However, sufficient economic data are still lacking to support the formulation of health policy and a particular challenge for the future is to conduct further health economic research on immunisation. Specific areas for such study include: effectiveness under field conditions (i.e., not under the conditions of a randomised controlled trial); the real value of economic production losses; the conditions for implementing novel immunization programmes; cost estimates for more ambitious immunization programmes; the economic benefits of combination vaccines. From this research, it will be important to disseminate the data and to adapt the findings to other countries. Nevertheless, the source of funding for research and its application in clinical trials programmes represent some of the practical problems faced by medical economics today within academia and the industry. PMID- 15755580 TI - Real world TB vaccines: clinical trials in TB-endemic regions. AB - The first 4 years of the 21st century have been an exciting time for tuberculosis (TB) research. After more than a half-century, the first new vaccines for TB are beginning to enter human clinical trials. Based on advances in proteomics and genomics, new vaccine candidates are filling the preclinical pipeline and our understanding of the natural history of M. tuberculosis infection is expanding rapidly. This review summarizes the developing consensus on the BCG vaccine and its limitations and discusses clinical trials of some of the new vaccines intended to supplement or replace it. PMID- 15755579 TI - Current progress in tuberculosis vaccine development. AB - The tuberculosis vaccine field has blossomed in the past 10 years, with over a hundred new candidates going through animal model testing, and several now entering or approaching clinical trial evaluation. In this brief review the current animal screening models are discussed, as are the various types of new vaccines that have been developed. New approaches, especially in the area of BCG boosting in various prime/boost protocols, are starting to show considerable promise. More sophisticated readouts, including imaging approaches such as magnetic resonance imaging, and better definition of the immunopathology of the lung disease process, should help accelerate vaccine development even further in the next decade. PMID- 15755581 TI - Do successful tuberculosis vaccines need to be immunoregulatory rather than merely Th1-boosting? AB - Tuberculosis vaccine candidates are entering clinical studies in areas where BCG fails. This is a high-risk strategy. We suggest that geographical variation in the efficacy of BCG is related to the presence in developing countries of a cross reactive background Th2-like response, probably attributable to exposure of mother and infant to helminths and environmental mycobacteria. Such Th2-like activity can stop Mycobacterium tuberculosis from being pushed into a latent state by the Th1 response, impair bactericidal functions and cause toxicity of TNF-alpha and pulmonary fibrosis. A successful vaccine, rather than driving a Th1 response, might need to suppress this pre-existing subversive Th2-like component. PMID- 15755582 TI - Developing an epitope-driven tuberculosis (TB) vaccine. AB - Epitope-driven vaccines are created from selected sub-sequences of proteins, or epitopes, derived by scanning the protein sequences of pathogens for patterns of amino acids that permit binding to human MHC molecules. We developed a prototype tuberculosis (TB) vaccine that contains epitopes derived by (1) EpiMer mapping of previously published secreted proteins derived from Mycobacterium tuberculosis (Mtb), and (2) EpiMatrix mapping of selected Mtb genome open reading frames (ORFs). Each of the epitopes contains at least three distinct class II MHC binding motif matches. These Mtb epitope selections were validated by measuring T cell responses from peripheral blood mononuclear cells (PBMC) obtained from healthy, asymptomatic tuberculin skin test-positive donors. Twenty-four validated Mtb epitopes were selected for inclusion in a DNA plasmid vector. We immunized HLA-DR B*0101 transgenic mice with this vaccine prototype augmented by co administration of rIL-15. Following administration of three immunizations at 14 day intervals in conjunction with rIL-15, epitope-specific T cell responses were observed to eight of the 24 epitopes contained in the DNA construct, one week following the last injection. The systematic application of bioinformatics tools to whole genomes, in combination with in vitro methods for screening and confirming epitopes, may lead to the development of novel vaccines for infectious diseases like TB. PMID- 15755583 TI - Novel recombinant BCG and DNA-vaccination against tuberculosis in a cynomolgus monkey model. AB - We have developed two novel tuberculosis (TB) vaccines: a DNA vaccine combination expressing mycobacterial heat shock protein 65 (Hsp65) and interleukin-12 (IL-12) by using the hemagglutinating virus of Japan (HVJ)-liposome (HSP65+IL-12/HVJ) and a recombinant BCG harboring the 72f fusion gene (72f rBCG). These vaccines provide remarkable protective efficacy in mouse and guinea pig models, as compared to the current by available BCG vaccine. In the present study, we extended our studies to a cynomolgus monkey model, which is currently the best animal model of human tuberculosis, to evaluate the HSP65+IL-12/HVJ and 72f rBCG vaccines. Vaccination with HSP65+IL-12/HVJ as well as 72f rBCG vaccines provided better protective efficacy as assessed by the Erythrocyte Sedimentation Rate, chest X-ray findings and immune responses than BCG. Most importantly, HSP65+IL 12/HVJ resulted in an increased survival for over a year. This is the first report of successful DNA vaccination and recombinant BCG vaccination against M. tuberculosis in the monkey model. PMID- 15755584 TI - HIV vaccine development by computer assisted design: the GAIA vaccine. AB - The design of epitope-driven vaccines that address the global variability of HIV has been significantly hampered by concerns about conservation of the vaccine epitopes across clades of HIV. We developed two computer-driven methods for improving epitope-driven HIV vaccines: the Epi-Assembler, which derives representative or "immunogenic consensus sequence" (ICS) epitopes from multiple viral variants, and VaccineCAD, which reduces junctional immunogenicity when epitopes are aligned in a string-of-beads format for insertion in a DNA expression vector. In this study, we report on 20 ICS HIV-1 peptides. The core 9 mer contained in these consensus peptides was conserved in 105-2250 individual HIV-1 strains. Nineteen of the 20 ICS epitopes (95%) evaluated in this study were confirmed in ELISpot assays using peripheral blood monocytes obtained from 13 healthy HIV-1 infected subjects. Twenty-five ICS peptides (all 20 of the peptides evaluated in this study and 5 additional ICS epitopes) were then aligned in a pseudoprotein string using "VaccineCAD", an epitope alignment tool that eliminates immunogenicity created by the junctions between the epitopes. Reordering the construct reduced the immunogenicity of the junctions between epitopes as measured by EpiMatrix, an epitope mapping algorithm. The reordered construct was also a more effective immunogen in vivo when tested in HLA-DR transgenic mice. These data confirm the utility of bioinformatics tools to design novel vaccines containing "immunogenic consensus sequence" T cell epitopes for a globally relevant vaccine against HIV. PMID- 15755585 TI - T-cell vaccination against anti-CD4 autoimmunity in HIV-1 subtypes B and C infected patients--an extended open trial. AB - This study is an extended clinical trial of the one initiated and reported in the Journal of Clinical Virology 2004;31S:S48-54. Thirteen HIV-1 patients (eight subtype B and five subtype C) that manifested T-cell autoimmunity to recombinant human CD4 (rCD4) were treated with T-cell vaccine composed of glutaraldehyde treated autologous anti-CD4 reactive T-cells and compared to historical seven non vaccinated HIV-1-infected subjects. This study proved to be feasible and safe. Follow-up study revealed that 7/8 subtype B and 2/4 subtype C patients (one has just received the first TCV injection) responded with a persistent increase in their blood CD4 T-cell levels and four subtype B patients manifested decreased anti-CD4 autoimmunity. Despite highly active antiretroviral therapy (HAART), the persistence of CD4 T-cell lymphopenia may be associated with anti-CD4 autoimmunity. T-cell vaccination (TCV) may decrease such autoimmunity and elevate CD4 T-cell numbers. PMID- 15755586 TI - Protection by dendritic cells-based HIV synthetic peptide cocktail vaccine: preclinical studies in the SHIV-rhesus model. AB - Vaccine development efforts against HIV-1 have been hindered because of the high mutation rate of the virus, and limitations for direct testing of HIV antigens in animal models to discern the nature of protective immunity. We developed a multivalent vaccine comprised of highly conserved HIV envelope peptide cocktail focused on priming antigen-specific helper T cell and CTL responses. Here we report protection of rhesus macaques against pathogenic SHIV(89.6P) challenge through priming cell-mediated immunity by prophylactic vaccination with the peptide-cocktail delivered by dendritic cells. Compared to monkeys mock vaccinated or immunized with the peptide cocktail using IFA, vaccination with peptide cocktail-pulsed DC showed significant protection from AIDS-associated mortality and reduction in plasma viremia to undetectable levels. PMID- 15755587 TI - Antigenic complementarity between HIV and other AIDS-associated infections results in idiotype-antiidiotype antibody complexes that cross react with lymphocyte proteins. AB - HIV proteins mimic HLA proteins, and the proteins of cofactor infections (cytomegalovirus, hepatitis viruses, mycobacteria, mycoplasmas) mimic CD4 proteins, making some HIV antigens molecularly complementary to cofactor antigens. Antibodies to HIV and its cofactors should therefore act like idiotype antiidiotype pairs. Over 2000 combinations of antibodies were tested for such complementarity using modified forms of Ouchterlony immunodiffusion and ELISA. HIV antibodies do precipitate some antibodies against cofactor infections including CMV, HBV, mycobacteria and mycoplasmas. These antibodies also mimic antibodies against HLA and CD4 proteins. Thus, some combinations of HIV with cofactor infections may induce lymphocytotoxic antibodies (LCTA), a risk that must be addressed in vaccine development. PMID- 15755588 TI - Evaluation of a synthetic vaccine construct as antigen for the detection of HIV induced humoral responses. AB - Synthetic V3 loop peptides hold numerous benefits for the development of HIV vaccines and in immunodiagnosis; however, their use is limited due to the extensive antigenic variability of this region. The effectiveness of a potential HIV vaccine component, which accounts for V3 loop variability, is evaluated here. A branched peptide construct representing multiple sequences and allowing 1.8 x 10(16) possible permutations was developed to mimic circulating HIV-1 subtype C, V3 loops. The construct was found to be immunogenic, able to induce neutralizing antibodies and sensitive to HIV-1 subtype B/C and HIV-2. This alternative antigen format also incorporates conformational epitopes. PMID- 15755589 TI - Targeting carbohydrate antigens in HIV vaccine development. AB - Peptide mimotopes provide a strategy to augment human immunodeficiency virus 1 (HIV-1) specific carbohydrate reactive immune responses. Their antigenic and immunological properties will depend on the optimization of motif clustering and multimerization. We observe that structural variants of the same mimetic motif, linear versus cyclic, can be used to tune the properties of the antibodies elicited. The expansion of the database of mimotope sequence motifs can be increased by analyzing structures that bind to HIV directed monoclonal antibody 2G12 and the lectin Concanavalin A (Con A), fostering new mimotope designs. Such analysis indicates that these reagents bind to subsets of mannosyl antigens on the envelope (env) protein. PMID- 15755590 TI - Vaccination against hepatitis A during outbreaks starting in schools: what can we learn from experiences in central Italy? AB - Two outbreaks of hepatitis A started almost simultaneously in a maternal school and in a day care centre located at opposite sides of Florence, Italy, at the end of 2002. Both of them originated from immigrant children, and in both cases, hepatitis A was initially not recognised due to aspecific symptoms. While vaccination of contacts started with delay in the first outbreak, the same intervention was organised and performed in 3 days in the other. The outbreak starting in the maternal school caused 30 notified cases, plus 7 cases diagnosed retrospectively. Nine of them were in a secondary school, where vaccination (in accordance with the Italian national guidelines on hepatitis A (HA) vaccination) had been started only after a secondary case occurred. Only three cases occurred overall in the other outbreak starting in the day care centre, where >80% of infants, children and personnel were immunised. Although few asymptomatic infections probably occurred, no source of contagion existed any longer 2 months after immunisation. A rapid vaccination of school and family contacts of hepatitis A cases after the first case (irrespective of school grade) seems to play an important role to shorten outbreak duration. PMID- 15755591 TI - Dramatic decline in acute hepatitis B infection and disease incidence rates among adolescents and young people after 12 years of a mass hepatitis B vaccination programme of pre-adolescents in the schools of Catalonia (Spain). AB - The aim of the study was to describe the impact of hepatitis B vaccination and disease incidence in adolescents and young people 12 years after the launching of a mass hepatitis B vaccination of pre-adolescents in schools. Vaccination coverage was assessed using administrative and serological data. Infection trends were evaluated by means of seroepidemiological surveys. High levels of vaccination coverage and vaccine-induced immunity were achieved. The resulting low proportions of susceptible adolescents and young people have undoubtedly contributed to the substantial reduction in the prevalence of hepatitis B infection in the 15-24 years age group (0.9 per 100 in 2001 versus 9.3 per 100 in 1986) and in the reported incidence of hepatitis B cases (80% reduction). Over the last 3 years, the declining trend seems to have been halted, although 35% of cases reported during this period corresponded to immigrants. PMID- 15755592 TI - Efficiency of the incorporation of the hepatitis A vaccine as a combined A+B vaccine to the hepatitis B vaccination programme of preadolescents in schools. AB - In 1998, the Department of Health of Catalonia (Spain) began universal vaccination of preadolescents against hepatitis A by replacing the simple hepatitis B vaccine with a combined hepatitis A+B vaccine. Economic analyses were made of the two alternative strategies: to continue with the simple hepatitis B vaccination or to replace the simple vaccine with a combined hepatitis A+B vaccine. The analysis was made from the societal perspective and the time horizon considered was 25 years. In the base case, (estimated annual hepatitis A incidence of 15 per 100,000 and incremental price of the hepatitis A+B vaccine over the simple hepatitis B vaccine of 1.98) the net present value of the programme was positive (+533,708) and the benefit-cost ratio was 2.58. If the estimated disease incidence were reduced by half, the programme would still be efficient. PMID- 15755593 TI - MeNZB: a safe and highly immunogenic tailor-made vaccine against the New Zealand Neisseria meningitidis serogroup B disease epidemic strain. AB - Clinical studies have been conducted in New Zealand evaluating the safety and immunogenicity of an outer membrane vesicle (OMV) vaccine, MeNZB, developed to control epidemic disease caused by group B meningococci, subtype P1.7b,4. MeNZB, administered in a three-dose regimen, was well tolerated and induced a seroresponse, defined as a four-fold rise (> or =titre 8) in serum bactericidal antibodies against the vaccine strain 4-6 weeks after the third vaccination, in 96% (95% confidence interval (CI): 79-100%) of adults, 76% (95% CI: 72-80%) of children, 75% (95% CI: 69-80%) of toddlers and 74% (95% CI: 67-80%) of infants receiving MeNZB. In conclusion, these findings suggest that MeNZB is safe and is likely to confer protection against systemic group B meningococcal disease caused by the epidemic strain. PMID- 15755594 TI - From secondary prevention to primary prevention: a unique strategy that gives hope to a country ravaged by meningococcal disease. AB - New Zealand has been affected by an epidemic of group B meningococcal disease dominated by a strain defined as, B:4:P1.7b,4. Over 5550 cases and 222 deaths have been reported since 1991 in a population of 4 million people. Meningococcal disease cases notified on EpiServ database operated by Institute of Environmental Science and Research Limited through to 30 September 2004. Through the collaborative efforts of a government agency, vaccine company, university and laboratory institute, clinical trials of the Chiron produced outer membrane vesicle (OMV) strain-specific MeNZB vaccine were run in rapid succession. The delivery of MeNZB will be New Zealand's largest immunisation programme with three doses given at 6-week intervals to over 1 million people aged 6 weeks-19 year olds inclusive. Planning, co-ordinating and delivering the immunisation programme is a challenging project for the New Zealand Health Sector. PMID- 15755595 TI - The concept of "tailor-made", protein-based, outer membrane vesicle vaccines against meningococcal disease. AB - Protein-based, outer membrane vesicle (OMV) vaccines have previously proven to be efficacious against serogroup B meningococcal disease in Norway and Cuba. Currently, a public health intervention is going on in order to control a serogroup B epidemic in New Zealand. The scale-up and standardization of vaccine production required for controlling the New Zealand epidemic has allowed the establishment of large-scale GMP manufacturing for OMV vaccines. The outcome of this will be licensing of the vaccine in New Zealand and possibly other countries. The availability of licensed OMV vaccines raises the question of whether such vaccines may provide the opportunity to control other outbreaks and epidemics. For instance, such a vaccine could control a localised outbreak of group B meningococci in Normandy, France. "Tailor-made" vaccines, focusing on the sub-capsular antigens may also be considered for use in sub-Saharan Africa for the prevention of the recurrent outbreaks by serogroups A and W135 meningococci. This assumption is based on the epidemiological observation that meningococcal outbreaks in Africa are clonal and are strikingly stable regarding their phenotypic characteristics. PMID- 15755596 TI - Outer membrane protein (OMP) based vaccine for Neisseria meningitidis serogroup B. AB - A family of outer membrane lipoproteins of Neisseria meningitidis, LP2086, has been shown to induce serum bactericidal activity against a broad variety of meningococcal strains. Two sub-families of serologically distinct LP2086 proteins (A and B) have been identified. In the present study, we have shown that polyclonal anti-serum against rLP2086 is protective in vivo in an infant rat passive-protection model. Additionally, the LP2086 protein is displayed on the surface of 91% meningococcal strains as measured in a whole cell ELISA using polyclonal anti-sera raised against these proteins. We also demonstrate based on the reactivity of anti-rLP2086 antibody with recombinantly expressed C- and N terminal fragments of rLP2086 in a Western blot assay that the C-terminal fragment of LP2086 dictates sub-family specificity and the N-terminal fragment determines the family specificity. A formulation containing family A and B of LP2086 potentially would provide broad protection against a majority of Neisseria meningitidis strains. PMID- 15755597 TI - The development of a meningococcal disease vaccine based on Neisseria lactamica outer membrane vesicles. AB - Serogroup B meningococcal disease remains a serious problem in many countries and no effective vaccine is currently available. Immunological and epidemiological evidence suggests that carriage of commensal Neisseria species is involved in the development of natural immunity against meningococcal disease. Neisseria lactamica has many surface structures in common with Neisseria meningitidis and may be the most important of these species. We have produced extensive pre clinical data, which indicate that N. lactamica outer membrane vesicles (OMVs) may provide a vaccine effective against diverse disease-causing meningococcal strains. Immunisation with N. lactamica OMVs protected against lethal challenge with diverse meningococcal isolates in a mouse intraperitoneal challenge model of meningococcal disease and we are developing this vaccine for use in a phase I safety and immunogenicity study in adult volunteers. We have shown that OMVs produced from bacteria grown under iron-limited or iron-rich conditions provide equivalent protection in the mouse infection model and thus OMVs produced from iron-rich will be used. Sterile filtration of N. lactamica OMVs has proved difficult but this has been improved by resuspending the vesicles in a buffer, which increases their surface zeta potential. The vaccine is currently being manufactured and validated ELISA protocols have been developed for the analysis of serological responses. PMID- 15755598 TI - Experimental disease models for the assessment of meningococcal vaccines. AB - Animal infection models are valuable for the development and preclinical assessment of meningococcal vaccines in the absence of clear in vitro correlates of protection for protein-based serogroup B vaccines. It is only in animal models that interactions of the organism with the innate, humoral and cellular immune systems can be assessed. However, humans are the only natural host for Neisseria meningitidis and there is no ideal disease model using laboratory animals that mimics the course of human disease. The two most widely used models are intraperitoneal (i.p.) infection of adult mice or infant rats. The mouse i.p. infection model requires an exogenous iron source (e.g. human transferrin) to obtain a lethal bacteraemic infection and can be used to assess both active and passive immunisation. The virulence of wild-type and knockout mutants can also be compared. i.p. infection of infant rats has been used to assess passive protection provided by sera raised against vaccine candidates or human vaccine sera. However, the duration of bacteraemia is short, mortality is low and active protection cannot be assessed. Recent developments using transgenic mice expressing human CD46 give hope that improved models will be developed. PMID- 15755599 TI - Validation of the serum bactericidal assay for measurement of functional antibodies against group B meningococci associated with vaccine trials. AB - Provisional licensure of the trial vaccine, MeNZB, required demonstration of immune responses in vaccines, as measured by a validated Serum Bactericidal Assay (SBA). Reported are the investigations undertaken to define test parameters, lower limits of quantitation and measurement of SBA reproducibility. Results helped to formulate the operating procedure for the measurement of serum bactericidal antibodies during six age-group MeNZB vaccine trials. The lower limit of quantitation was determined as a titre of 4. A four-fold rise in antibody (sero-conversion) from a pre-vaccination titre of 2 (<4) required a minimum post-vaccination titre of 8, a more stringent measurement than has been used in other published studies. PMID- 15755600 TI - Meningococcal surrogates of protection--serum bactericidal antibody activity. AB - Despite the availability of anti-microbial agents effective against Neisseria meningitidis, meningococcal disease continues to be a major global health problem, particularly in the very young. Serogroup A meningococci cause large epidemics in sub-Saharan Africa, whilst serogroups B and C organisms are responsible for sporadic cases and localised outbreaks of disease world-wide. For measuring functional activity, the serum bactericidal antibody (SBA) assay is the most important method. It is mediated by antibody and complement resulting in lysis of the bacterial cells. To date the SBA has proved to be the best surrogate of protection for all serogroups. For serogroup C, an SBA titre of either >/ or =4 or > or =8 has being utilised for putatively indicating protection when using either human or baby rabbit complement, respectively. For serogroup B, the proportions of vaccines with > or =4-fold rises in SBA pre- to post-vaccination or SBA titres > or =4 have been correlated with clinical efficacy in trials of outer membrane vesicle (OMV) vaccines in Cuba, Brazil and Norway. SBA activity as a correlate of protection for evaluating the immune response to meningococcal vaccines is described in this review. PMID- 15755601 TI - New Zealand's epidemic of meningococcal disease described using molecular analysis: implications for vaccine delivery. AB - New Zealand's epidemic of meningococcal disease began in mid 1991. Surveillance of meningococcal disease in New Zealand is based on a combination of disease notification and organism characterisation. Case numbers and population rates rose from 53 (1.5 per 100,000 population) in 1990 to a high of 650 (17.4 per 100,000 population) in 2001. The highest rates of disease occur in Pacific peoples under 20 years but the highest percentages of cases occur in Maori and European New Zealanders. The epidemic has been driven by a strain identified as B:4:P1.7b,4, ST-41/44 complex/Lineage III. The stability of the P1.7b,4 Por A protein has been demonstrated suggesting that the epidemic may be controlled by a strain-specific OMV vaccine. PMID- 15755602 TI - Evaluating the post-licensure effectiveness of a group B meningococcal vaccine in New Zealand: a multi-faceted strategy. AB - A nationwide strategy to control a group B meningococcal disease epidemic in New Zealand using an epidemic strain-specific vaccine (MeNZB ) commenced in 2004. In the absence of randomised controlled trials investigating the efficacy of this particular vaccine, a complement of observational methods are planned to evaluate the post-licensure effectiveness of this vaccine strategy. The two main approaches involve a Poisson regression model investigating the overall impact of the MeNZB programme on disease rates over time capitalising on detailed population-based disease surveillance data and the staged roll-out of the vaccine campaign, and a case-control study that aims to estimate vaccine effectiveness in pre-school children. The studies are designed to minimise the potential biases inherent in all observational methods and provide critical data on the effectiveness of a major public health intervention. PMID- 15755603 TI - Malaria vaccines: using models of immunity and functional genomics tools to accelerate the development of vaccines against Plasmodium falciparum. AB - Naturally acquired immunity and immunity acquired after immunization with attenuated parasites indicate that a vaccine against malaria is feasible. Several obstacles have stymied malaria vaccine development, among them our poor understanding of protective immunity and technical difficulties for studying gene and protein expression in the Plasmodium falciparum parasite. Pregnancy malaria offers a model approach for vaccine development: recent findings have elucidated the basis for disease pathogenesis and protective immunity in this syndrome, and this understanding has focused the effort to identify the optimal antigens for a pregnancy malaria vaccine. In parallel, functional genomics tools are overcoming several of the obstacles for studying protein expression in the malaria parasite, vastly accelerating the pace for antigen discovery. Together, these conceptual and technological advances allow a rational approach to vaccine antigen selection, in which a finite number of antigens are selected from the entire genome by merit of the expression patterns and specific features. These candidate antigens are then subjected to detailed studies according to criteria established by the understanding of pathogenesis and protective immunity, to identify the optimal antigens for inclusion in subunit vaccines. PMID- 15755605 TI - Requiring influenza vaccination for health care workers: seven truths we must accept. AB - In this paper we outline the seven primary truths supporting the call for requiring influenza immunization of all health care workers. We view this as a serious patient safety issue, given the clear and compelling data regarding the frequency and severity of influenza infection. In addition, clear-cut safety, efficacy, economic, legal, and ethical platforms support the use of influenza vaccine. Unfortunately health care workers have demonstrated, over almost 25 years that they are unwilling to comply with voluntary influenza immunization programs utilizing a variety of education and incentive programs, at rates sufficient to protect the patients in their care. We suggest that an annual influenza immunization should be required for every health care worker with direct patient contact, unless a medical contraindication or religious objection exists, or an informed declination is signed by the health care worker. High rates of health care worker immunization will benefit patients, health care workers, their families and employers, and the communities within which they work and live. PMID- 15755606 TI - Policy evaluation for the subsidy for influenza vaccination in elderly. AB - OBJECTIVE: In Japan, the subsidy of influenza vaccination for the elderly was introduced in November 2001. This paper examines its policy evaluation from the viewpoint of cost-benefit analysis. MATERIALS: The data of copayment of influenza vaccination, population and shot rate of the elderly are surveyed by telephone interview to the correspondents in the local governments of Tokyo metropolitan and other 12 big cities in Japan. The mortality due to pneumonia or influenza is obtained from Vital Statistics of Population. METHOD: At first, I examine the impact of amount of copayment, through its effect on shot rate, on the percentage of elderly receiving influenza vaccinations. Using these estimation results, benefit-cost ratio (BCR) is calculated. RESULTS: The estimated coefficient of copayment on shot rate is -0.007 and statistically significant. Shot rate significantly reduces pneumonia and influenza mortality and its magnitude is 0.0028. The obtained net benefit (NB) is 134.9 million yen or US$1.08 billion and benefit-cost ratio is 22.9 and its 95% confidence interval is [2.2, 43.7]. DISCUSSION: If copayment would be cut by a 1000 yen (US$8), it could avoid about 400 deaths in average big city. The benefit-cost ratio is quite high compared with the other countries or other vaccinations. CONCLUSION: We found the strong evidence in a sense of cost-benefit analysis in the subsidy for influenza vaccination in the elderly. PMID- 15755604 TI - Towards an RTS,S-based, multi-stage, multi-antigen vaccine against falciparum malaria: progress at the Walter Reed Army Institute of Research. AB - The goal of the Malaria Vaccine Program at the Walter Reed Army Institute of Research (WRAIR) is to develop a licensed multi-antigen, multi-stage vaccine against Plasmodium falciparum able to prevent all symptomatic manifestations of malaria by preventing parasitemia. A secondary goal is to limit disease in vaccinees that do develop malaria. Malaria prevention will be achieved by inducing humoral and cellular immunity against the pre-erythrocytic circumsporozoite protein (CSP) and the liver stage antigen-1 (LSA-1). The strategy to limit disease will target immune responses against one or more blood stage antigens, merozoite surface protein-1 (MSP-1) and apical merozoite antigen 1 (AMA-1). The induction of T- and B-cell memory to achieve a sustained vaccine response may additionally require immunization with an adenovirus vector such as adenovirus serotype 35. RTS,S, a CSP-derived antigen developed by GlaxoSmithKline Biologicals in collaboration with the Walter Reed Army Institute of Research over the past 17 years, is the cornerstone of our program. RTS,S formulated in AS02A (a GSK proprietary formulation) is the only vaccine candidate shown in field trials to prevent malaria and, in one instance, to limit disease severity. Our vaccine development plan requires proof of an individual antigen's efficacy in a Phase 2 laboratory challenge or field trial prior to its integration into an RTS,S-based, multi-antigen vaccine. Progress has been accelerated through extensive partnerships with industrial, academic, governmental, and non governmental organizations. Recent safety, immunogenicity, and efficacy trials in the US and Africa are presented, as well as plans for the development of a multi antigen vaccine. PMID- 15755608 TI - Prophylaxis of acute respiratory virus infections using nucleic acid-based drugs. AB - Acute respiratory virus infections such as SARS and pandemic influenza are highly contagious diseases that cause global crisis, and inflict severe human mortality and morbidity. Vaccines against these viruses are either unavailable or do not provide adequate protection. In the absence of effective vaccines, nucleic acid based immunomodulators have the potential to offer effective, broad-spectrum protection against these deadly pathogens. Poly ICLC and CpG oligonucleotides are promising gene-based drugs which have been shown in animal studies to protect against acute respiratory virus infections. Poly ICLC is a synthetic double stranded RNA (dsRNA), and an effective interferon-inducer and natural killer cell activator. When encapsulated in liposomes, poly ICLC offers complete protection (100% survival rate in pretreated group versus 0% survival in control group) against a lethal respiratory challenge of influenza A virus in mice. This antiviral effect has been shown to persist for up to 3 weeks post-drug treatment. Poly ICLC pretreatment also protects mice against a respiratory challenge of western equine encephalitis (WEE) virus, at a level comparable to inactivated WEE vaccine. CpG oligos in liposomes also provided high level of protection against the lethal influenza challenge. Together, these studies suggest nucleic acid based immunomodulators are promising antiviral agents which can offer effective and non-specific protection against acute respiratory virus infections. PMID- 15755607 TI - Peptide-based candidate vaccine against respiratory syncytial virus. AB - We engineered a 21-mer peptide representing amino acids 170-190 of the respiratory syncytial virus (RSV) G protein as a fusion with the Alfalfa mosaic virus (AlMV) coat protein (CP), produced recombinant AlMV particles presenting this peptide (VMR-RSV) on their surfaces and tested the immunogenicity in vitro in human dendritic cells and in vivo in non-human primates. Significant pathogen specific immune responses were generated in both systems: (i) human dendritic cells armed with VMR-RSV generated vigorous CD4+ and CD8+ T cell responses; (ii) non-human primates that received these particles responded by mounting strong cellular and humoral immune responses. This approach may validate the use of a novel RSV vaccine delivery vehicle in humans. PMID- 15755609 TI - The development of vaccines against SARS corona virus in mice and SCID-PBL/hu mice. AB - We have investigated to develop novel vaccines against SARS CoV using cDNA constructs encoding the structural antigen; spike protein (S), membrane protein (M), envelope protein (E), or nucleocapsid (N) protein, derived from SARS CoV. Mice vaccinated with SARS-N or -M DNA using pcDNA 3.1(+) plasmid vector showed T cell immune responses (CTL induction and proliferation) against N or M protein, respectively. CTL responses were also detected to SARS DNA-transfected type II alveolar epithelial cells (T7 cell clone), which are thought to be initial target cells for SARS virus infection in human. To determine whether these DNA vaccines could induce T cell immune responses in humans as well as in mice, SCID-PBL/hu mice was immunized with these DNA vaccines. As expected, virus-specific CTL responses and T cell proliferation were induced from human T cells. SARS-N and SARS-M DNA vaccines and SCID-PBL/hu mouse model will be important in the development of protective vaccines. PMID- 15755610 TI - Evaluation of modified vaccinia virus Ankara based recombinant SARS vaccine in ferrets. AB - Severe acute respiratory syndrome (SARS) caused by a newly identified coronavirus (SARS-CoV) remains a threat to cause epidemics as evidenced by recent sporadic cases in China. In this communication, we evaluated the efficacy and safety of two SARS vaccine candidates based on the recombinant modified vaccinia Ankara (MVA) expressing SARS-CoV spike or nucleocapsid proteins in ferrets. No clinical signs were observed in all the ferrets challenged with SARS-CoV. On the other hand, vaccination did not prevent SARS-CoV infection in ferrets. In contrast, immunized ferrets (particularly those immunized with rMVA-spike) exhibited significantly stronger inflammatory responses and focal necrosis in liver tissue after SARS-CoV challenge than control animals. Thus, our data suggest that enhanced hepatitis is linked to vaccination with rMVA expressing SARS-CoV antigens. PMID- 15755611 TI - Efficacy of DNA vaccination against western equine encephalitis virus infection. AB - The efficacy of a DNA vaccine against western equine encephalitis (WEE) infection in mice was evaluated. The 26S structural region was expressed, in vitro from an internal T7 promoter using a rabbit reticulysate transcription/translation system; and from a CMV promoter after transfection into Vero cell monolayers. The proteins synthesized were reactive with anti-WEE virus (WEEV) antibodies, both in western blot analysis and histochemical staining, respectively. When the DNA vaccine plasmid, pVHX-6, was administered intraepidermally to mice, followed by challenge in a lethal mouse model, the level of protection obtained ranged from 50 to 100% amongst three strains of WEEV. Preliminary results suggest the protective immunity provided by the DNA vaccine appears to be a cell-mediated immune response, as elevated cytotoxic T lymphocyte activity was detected against the E2 protein in a T-cell proliferation assay. The efficacy results suggest a DNA vaccine may be a promising approach against WEE infection. PMID- 15755612 TI - Rabies control in South and Southeast Asia. AB - We have the knowledge and tools to eliminate the threat of canine rabies but this disease, nevertheless, remains a public health threat in many parts of the world. Lack of motivation by governments, cultural issues and inadequate funding remain barriers. This is amazing since the number of human rabies deaths worldwide is greater than that from polio, meningococcal meningitis, Japanese encephalitis, yellow fever, SARS, bird flue and other scourges that attract more attention. Safe and effective vaccines are now widely available. Reduced dose effective and less expensive post-exposure vaccination regimens have helped eliminate nerve tissue vaccines in Thailand, Philippines and Sri Lanka. India and Pakistan, the major users of dangerous nerve tissue derived Semple type vaccine, are now considering following suite. Immediate wound care and prompt use of a potent vaccine will save a majority of infected persons. Rabies immunoglobulin, injected into and around bite wounds, provides added safety for the severely exposed. The high cost of rabies immunoglobulin and tissue culture vaccines are remaining barriers, but new manufacturers and the use of intradermal vaccination schedules can reduce costs. Ultimately, it is the need to control rabies in dogs that must occupy most of our attention. The tools are available, but attitudes must change before they can be applied. There have been many new developments since publication of the last WHO rabies expert committee report in 1992 (new version in print)] and we will address those that have practical applicability. PMID- 15755613 TI - Induction of immune responses and partial protection in mice after skin immunization with rotavirus VP6 protein and the adjuvant LT(R192G). AB - Oral or intranasal administration of mice with rotavirus VP6/LT(R192G) vaccine induces between 95 and 99% protection against fecal shedding of rotavirus after challenge. However, mucosal administration of LT(R192G) is controversial. Subcutaneous, intradermal or Biojector injection induced high titers of serum VP6 specific IgG, eliciting only partial to no protection (73, 0 and 26%, respectively), while transcutaneous delivery using gauze pad induced both poor immune responses and no protection (13%). A mixture of VP6-derived synthetic peptides induced >97, 48 and 33% protection after intranasal, gauze pad or Biojector administration, respectively. For needle-free delivery methods to be viable, improvements to these methods must be made to enhance the efficacy of the VP6 vaccine. PMID- 15755614 TI - Expression of the synthetic neutralizing epitope gene of porcine epidemic diarrhea virus in tobacco plants without nicotine. AB - Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in pigs, leading to death with a high mortality. In this study, the synthetic neutralizing epitope gene of PEDV, which was optimized based on the coding sequence of tobacco plant genes being modified, was expressed in the no-nicotine tobacco plants. The synthetic gene was cloned into the plant expression vector under the control of CaMV 35S promoter and transformed by an Agrobacterium mediated transformation. The amount of synthetic epitope protein of PEDV detected in the transgenic tobacco plants was approximately 2.1% of the total soluble plant protein, which was approximately five-fold higher than that expressed with the native gene. PMID- 15755615 TI - Protection against group A streptococcal infection by vaccination with self adjuvanting lipid core M protein peptides. AB - We have investigated the lipid polylysine core peptide (LCP) system as a self adjuvanting group A streptococcal (GAS) vaccine delivery approach. LCP constructs were synthesised incorporating peptides from the M protein conserved carboxy terminal C-repeat region, the amino terminal type-specific region and from both of these regions. Immunisation with the constructs without adjuvant led to the induction of peptide-specific serum IgG antibody responses, heterologous opsonic antibodies, and complete protection from GAS infection. These data indicate that protective immunity to GAS infection can be evoked using the self-adjuvanting LCP system, and point to the potential application of this system in human mucosal GAS vaccine development. PMID- 15755616 TI - Bacterial otitis media: a vaccine preventable disease? AB - Otitis media (OM) is the most common childhood illness for which medical advice is sought. Whilst the disease rarely results in death, there is a significant level of morbidity and economic burden on the community. Although the causes of OM are multifactoral, bacterial and viral infections are the single most important cause. Bacteria responsible for infections of the middle ear are predominantly, nontypeable Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis. Antibiotics have been widely used to treat children who present to a medical clinic with OM. However, given the high prevalence of this disease and the increasing incidence of microbial resistance to antibiotics, there is a need to develop alternative therapeutic strategies such as vaccination. Pneumococcal polysaccharide vaccination has produced disappointing results for effectiveness in preventing OM and there is evidence of an increased incidence of disease due to non-vaccine serotypes. An efficacious vaccine for bacterial OM would require combining protective protein antigens from all three causative bacteria. A combined bacterial-viral vaccine formulation would produce the most profound and sustained impact on reducing the global incidence of OM. PMID- 15755617 TI - United States Pharmacopeia activities in the area of vaccines, virology and immunology. AB - The United States Pharmacopeia (USP) develops public standards for medical products that are enforceable by FDA. USP general information chapters have been providing industrial and academic researchers alike with crucial guidance especially in areas where there is absence of regulatory guidance. In an effort to meet the challenge of rapid advances in vaccine research and manufacturing, the Council of Experts Committee for Vaccines, Virology, and Immunology of the US Pharmacopeia has recently initiated two new general chapters to provide advice for researchers and manufacturers in the vaccine and virology fields and beyond. Chapter 1235 Vaccines and Vaccine Test Methods will focus on manufacturing and analytical requirements for the different types of vaccines currently in manufacture and development. Chapter 1237 Virology Test Methods will discuss modern diagnostic virology techniques and a variety of tests as applicable to vaccine and biologics manufacturing. PMID- 15755618 TI - Studies of equivalence in clinical vaccine research. AB - The development of combination vaccines as well as the improved manufacture of vaccines have resulted in the need for clinical studies seeking equivalence or non-inferiority as investigators seek to demonstrate that combination vaccines achieve the same levels of efficacy, immunogenicity, and safety as their individual counterparts. Given the nature of the statistical analysis, studies of equivalence require particular attention to sample size. The current double significance method of Neyman-Pearson attempts to address problems with equivalence testing, but it leads to very large sample sizes and illogical results. With such large samples, one can find a clinically trivial difference that is still statistically significant. The late Professor Alvan R. Feinstein proposed a more logical approach that would call for smaller, more workable sample sizes. Understanding the issues involved in sample size calculations for such studies is important to those who design clinical vaccine studies. The implications of the calculations will have far-reaching effects on the feasibility of the study such as availability of subjects, the success with recruitment, and the overall expenses. In fact, the feasibility issues may prevent the study being undertaken at all. PMID- 15755619 TI - Sample sizes and negative studies in clinical vaccine research. AB - Negative studies of vaccine adverse events occur with some frequency in the published literature. They serve important roles in fending off claims posed by the anti-vaccine movement and in reinforcing public health efforts to prevent diseases. Still, negative studies frequently suffer from concerns of adequate sample size. The double-significance Neyman-Pearson formula for sample size calculation that is now in vogue results in immense sample sizes that can lead to illogical interpretations. The late Professor Alvan R. Feinstein, the father of quantitative clinical epidemiology, proposed a more logical approach that would reduce the confusion and call for more moderate sample sizes. Understanding the issues involved in sample size calculations is important to those who design clinical vaccine studies. The implications of the calculations have far-reaching effects upon elements of feasibility such as the expense of the study, the ability to recruit adequate numbers, and even whether the study will be done at all. PMID- 15755620 TI - An ingenious design for peptide vaccines. AB - For humoral immunization, it may be possible to make effective and safe peptide vaccines for various diseases by selection of proper B-cell epitopes. However, a lack of T-cell epitopes on short peptides, such as those associated with major histocompatibility complex (MHC)-restriction, is a major problem for peptide vaccine development. We propose a solution for the design of peptide vaccines that involves induction of broadly reactive T-cell epitopes via agretopes. The strategy involves positioning multi-agretope type peptides on the N-terminal side of a di-lysine linker and B-cell epitopes on the C-terminal side. The addition of the arginine-glysine-aspartate (RGD)-motif to the N terminus of the peptide enhances its immunogenicity, and enables nasal immunization without adjuvants. PMID- 15755621 TI - Generation of anti-beta-amyloid antibodies via phage display technology towards Alzheimer's disease vaccination. AB - The pathology of Alzheimer's disease (AD) shows a significant correlation between beta-amyloid peptide (betaAP) deposition and the clinical severity of dementia. The ability of site-directed antibodies towards the N-terminal region of beta amyloid peptide to suppress in vitro formation of toxic beta-amyloid serves as a factual basis for in vivo investigations. We localized the epitope of these anti aggregating antibodies, and injection of phage displaying this epitope induced antibodies against the whole anti-beta-amyloid peptide. In Alzheimer's diseased transgenic mice, these antibodies are delivered from the periphery to the CNS preventing beta-amyloid formation and/or dissolving such aggregates. Performance of such antigens opens up possibilities for development of an efficient, long lasting immunization procedure for treatment of Alzheimer's disease. PMID- 15755622 TI - General strategies for efficient adjuvant incorporation of recombinant subunit immunogens. AB - We have previously reported strategies for Escherichia coli production of recombinant immunogens fused to hydrophobic peptides or lipid tags to improve their capacity to be incorporated into an adjuvant formulation, e.g., immunostimulating complexes (iscoms). Recently, we also explored the strong interaction between biotin and streptavidin to achieve iscom association of recombinant immunogens. Plasmodium falciparum,Toxoplasma gondii and Neospora caninum antigens have served as model immunogens in the different studies. Generated fusion proteins have been found to be successfully incorporated into iscoms and high-titer antigen-specific antibody responses have been obtained upon immunization of mice. We believe that the different concepts presented, utilizing either hydrophobic peptide or lipid tags, or the recently explored biotin streptavidin principle, offer convenient methods to achieve efficient adjuvant incorporation of recombinant immunogens. PMID- 15755623 TI - Linkage of Ii-Key segment to gp100(46-58) epitope enhances the production of epitope-specific antibodies. AB - Linkage of the Ii-Key segment of the Ii protein to MHC class II epitope gp100(46 58) using a polymethylene linker significantly enhances the production of epitope specific antibodies in HLA-DR4-IE transgenic mice. This enhancement is not restricted by the spacer length in between the Ii-Key and epitope. The use of either IFA or CFA induced only epitope-specific IgG1. In contrast, CpG adjuvant induced both IgG1 and IgG2a isotypes. These results indicate that the Ii-Key hybrid technology is a novel and potent method to increase the immunogenicity of a MHC class II epitope. It can also be used to more efficiently generate epitope specific antibodies. PMID- 15755624 TI - Fusing subunit antigens to interleukin-2 and encapsulating them in liposomes improves their antigenicity but not their protective efficacy. AB - Subunit vaccines commonly lack sufficient immunogenicity to stimulate a comprehensive protective immune response in vivo. We have investigated the potential of specific cytokines (interleukin-2) and particulate delivery systems (liposomes) to enhance antigenicity. Here we report that the IgG1 and IFN-gamma responses to a subunit antigen, consisting of a T and B-cell epitope from Influenza haemagglutinin, can be improved when it is both fused to interelukin-2 and encapsulated in liposomes. However, this vaccine formulation was not able to protect animals against a challenge with live Influenza A/PR/8/34 virus. The addition of more potent immune stimulators may be necessary to improve responses. PMID- 15755625 TI - Cost-benefit and cost-effectiveness of the incorporation of the pneumococcal 7 valent conjugated vaccine in the routine vaccination schedule of Catalonia (Spain). AB - The health and economic costs and benefits of vaccinating a cohort of 60,000 children born in Catalonia in the year 2000 with the pneumococcal 7-valent conjugated vaccine were compared with the alternative of not implementing the vaccination programme. The time horizon fixed for the programme was 10 years for invasive disease, 2 years for all episodes of pneumonia and otitis media and 3.5 years for the placement of tympanostomy tubes. In the base case (incidence rate of invasive disease of 160 per 100,000 and price of the vaccine 50 euros) the net present value was negative, both from the societal perspective (-5.1million euros) and from the provider's perspective (-9.2million euros). The benefit-cost ratio was 0.59 euros from the societal perspective. The cost per disability adjusted life year (DALY) gained was 44,307 euros from the societal perspective and 80,291 euros from the provider's perspective. PMID- 15755626 TI - Surveillance system of vaccine adverse events and local data analysis--the experience in a middle-sized city in Brazil, 1999-2001. AB - We reviewed all vaccine adverse events (VAE) notified in a middle-sized Brazilian city (n=247) to the National Immunisation Program between January 1999 and December 2001. Vaccine doses used in that period were considered for rate estimates. Aspects of the surveillance system (SS) and their influences on collected data were considered, searching for contributions of local data analysis to investigation of VAE and to the monitoring of vaccine safety. Notification rates in our study were higher when compared to national data. Changes in the notification pattern were observed following vaccination campaign periods. An increase in aseptic meningitis cases temporally associated to yellow fever vaccine was detected. The analysis of local data provided information unperceived in national consolidated data. Through this analysis we detected: events related to application technique and handling; people's perception changes on VAE; and the local SS's ability to raise new hypothesis. We suggested changes to the notification form regarding data entry criteria and analysis. PMID- 15755627 TI - The "Green Channel" of the Veneto region as a model for vaccine safety monitoring in Italy. AB - Modern vaccinology and public health organizations need to satisfy an increased safety demand. Therefore, to improve adverse events following immunization (AEFIs) surveillance systems, some countries have established clinical evaluation centers for AEFI assessment and management of at risk individuals. In the Veneto region of Italy, the Green Channel operates through a counselling service for subjects with prior AEFI or with suspected contraindications to vaccine administration, and a surveillance system of the AEFIs reported in the region. Updated data on 753 consultations and 3023 AEFI analyses are discussed together with the opportunity to include the Green Channel model as part of an international vaccine safety network. PMID- 15755628 TI - Cancer vaccines and immunotherapies: emerging perspectives. AB - Research efforts over the last two decades studying immune responses to human carcinomas have demonstrated that antigens expressed by tumor cells can elicit specific cellular and humoral immune responses. Unfortunately, despite the observation that existent immune responses to these antigens are present in some patients with cancer, the tumors continue to progress. Thus, there has been considerable interest to augment these immune responses by immunization. Some of the clinical trials of the first cancer vaccines have provided evidence of clinical benefit thus encouraging the development of other vaccines. Challenges to development of such cancer vaccines include the identification and characterization of the antigen(s) to be targeted, the definition of the desired immune response to be elicited by the vaccine, and the choice of the appropriate vaccine delivery system. PMID- 15755629 TI - Therapeutic vaccines: a solution to the prevention problem? AB - Therapeutic vaccines are likely to provide a significant and growing contribution to tackling some of the manifold health problems that face us in the future. However, despite these real benefits, I suggest the need for caution in relation to one possible ethical argument that could be used to support the introduction of therapeutic vaccination over preventive vaccination for contagious diseases where both types of vaccination exist. I argue that therapeutic vaccination will not solve what I call the 'prevention problem': that is, a perceived injustice in the distribution of benefits and harms arising from population-based preventive vaccination policies. PMID- 15755630 TI - Tumour-dendritic hybrid cell vaccination for the treatment of patients with malignant melanoma: immunological effects and clinical results. AB - Hybrid cell vaccines of autologous tumour cells fused with allogenic dendritic cells (DC) combine the tumour's antigenicity with the immune-stimulatory capacity of mature dendritic cells and allogenic MHC class II molecules to activate T cell help and induce tumour-specific cytotoxic T cells. This concept was tested in a clinical trial with melanoma stage III and IV patients. Seventeen patients were evaluated: one experienced complete, one partial response and six stable disease with long survival times. Eleven of fourteen patients, clinical responders and non-responders alike, mounted high-frequency T cell responses to various tumour associated antigens. Failing clinical responses correlated with loss of antigenicity. PMID- 15755631 TI - The rationale for combined chemo/immunotherapy using a Toll-like receptor 3 (TLR3) agonist and tumour-derived exosomes in advanced ovarian cancer. AB - A clinical trial employing an immunotherapeutic approach based on the use of a Toll-like receptor 3 (TLR3) agonist and tumour-derived exosomes carrying tumour associated antigens is planned in advanced ovarian cancer in conjunction with conventional first line chemotherapy. Most patients with ovarian cancer present with advanced disease and despite high initial response rate to chemotherapy the majority will relapse within 2 years with poor overall survival. Tumour antigen specific T cells are naturally occurring in ovarian cancer patients and T cell infiltration of the tumour is highly prognostic. Novel immunotherapy to expand and activate tumour antigen-specific T cells combined with adjuvant treatment to overcome tumour-induced immunosuppression is considered to be therapeutically beneficial. The rationale for adopting such a combined approach is discussed here. PMID- 15755633 TI - Vaccination against human papillomavirus infection: a new paradigm in cervical cancer control. AB - Universal deployment of organized or opportunistic screening with Pap cytology in high and middle income countries has been the primary reason for the substantial reductions in cervical cancer morbidity and mortality during the last 50 years. However, in many low income countries Pap cytology screening is yet to be effectively implemented or has failed to reduce cervical cancer rates to an appreciable extent. Cervical cancer thus remains a critical public health problem that is second only to breast cancer in overall disease burden for women throughout the world. The fact that infection with certain human papillomavirus (HPV) types is now recognized as a necessary cause of this disease has led to new research fronts on the prevention of cervical cancer. Recent research on the safety and efficacy of candidate prophylactic vaccines against HPV have shown very promising results with nearly 100% efficacy in preventing the development of persistent infections and cervical precancerous lesions. Ongoing clinical studies are expected to provide further evidence of efficacy and will form the basis for licensing of candidate vaccines by the major pharmaceutical companies within 3-6 years. Although the future seems bright on the HPV vaccine front policy makers are strongly cautioned to avoid scaling back cervical cancer screening. It will take many years before we can rationally develop cervical cancer screening strategies that will be cost-effective for the proper surveillance of women protected by HPV vaccination. PMID- 15755632 TI - Immunotherapy with autologous tumor cell-BCG vaccine in patients with colon cancer: a prospective study of medical and economic benefits. AB - We have completed a multicenter, randomized controlled phase III clinical trial in Stages II and III colon cancer patients with active specific immunotherapy (ASI) using autologous tumor cells with an immunomodulating adjuvant bacillus Callmette-Guerin (BCG) vaccine (OncoVAX) in an adjuvant setting. In this study, patients were randomized to receive either OncoVAX therapy or no therapy after surgical resection of the primary tumor and stratified by stage of disease. Since the biologic essence of the effective tumor immunotherapy is the presence in the vaccine of a minimum number of viable, metabolically active, autologous tumor cells, the processing of the vaccine product, occurred within 48 h after surgery. Analysis of prognostic benefit in the pivotal phase III trial, with a 5.8 year median follow-up, showed that a beneficial effect of OncoVAX is statistically significant for all endpoints including recurrence-free interval, overall survival, and recurrence-free survival in Stage II colon cancer patients. Surgery alone cures approximately 65% of Stage II (Dukes B2, B3) colon cancer patients. In the remaining patients, OncoVAX in an adjuvant setting, significantly prolongs recurrence-free interval (57.1% relative risk reduction) and significantly improves 5-year overall survival and recurrence-free survival. No statistically significant prognostic benefits were achieved in Stage III (Duke's C1-C3) patients. A health economics assessment was performed on these results in Stage II colon cancer patients using disease-free survival and overall survival (for the entire intent-to-treat population). Cost-effectiveness, cost-utility and sensitivity analysis were applied with, cost of life years, recurrence-free life years and quality adjusted life years (QALYs) as the primary endpoints to this analysis. The perspective of the economic analysis was the current direct medical cost established by the health care providers. The introduction of new technologies often leads to additional costs. This report verified that the use of OncoVAX for patients with Stage II colon cancer not only has significant prognostic benefit and positive clinical outcomes, but also showed that OncoVAX therapy yields impressive health economics benefits. PMID- 15755634 TI - Development of a therapeutic vaccine for HSV-2. AB - A therapeutic vaccine for genital herpes is clearly needed. Vaccines recently developed for HSV-2 in humans have been aimed at eliciting CD4+ T cell responses and neutralizing antibody responses to two HSV-2 glycoproteins (gB and gD). These vaccines have had no therapeutic effect against HSV-2 in human clinical trials. To enable development of an efficacious vaccine, Corixa Corporation has made a major effort to identify novel antigens that can be recognized by human HSV-2 specific CD8+ and CD4+ T cells. Corixa Corporation's proprietary adjuvants and delivery systems, when combined with appropriate antigens, may allow the development of an effective therapeutic vaccine for HSV-2. PMID- 15755635 TI - Assessment of immunological competence and SV40 specific recall immunity in malignant pleural mesothelioma. AB - Diffuse malignant pleural mesothelioma (MPM) is the third most common lung malignancy showing rising incidence with 250,000 deaths expected from it in Western Europe over the next 35 year. The tumour is generally resistant to conventional treatment and there is urgent need for novel preventative and therapeutic measures to combat this growing public threat. Finding of SV40 DNA sequences in a high proportion (40-90%) of several series of MPM cases, and suggestion of its potential co-carcinogen role provide a rationale for the development of novel anti-MPM vaccines incorporating SV40 gene sequences or antigenic determinants. As a prelude to adopting this approach, general T cell function was examined in relatively early cases of MPM presenting for biopsy or debulking surgery. CD8+ T cell responses were studied using antigenic epitopes of common viral antigens covering a broad range of haplotypes. 74.1% (20/27) of MPM patients and 80% (8/10) of the control subjects showed T cell responsiveness to the viral peptides mix, whilst a small proportion showed SV40 specific recall immunity. PMID- 15755636 TI - Immunopropylactic approaches against chemical carcinogenesis. AB - Immunoprophylactic strategies using carcinogen-specific antibodies have so far received little attention. This may be due to experimental difficulties of in vivo chemical carcinogenesis models, which conveniently use excessive doses of carcinogen, which do not reflect environmental exposure and which cannot be matched by molar equivalents of antibodies. However, more recent studies have now demonstrated that both mucosal and systemic antibodies may afford protection against low doses of environmental carcinogens, at least when stoichiometry between carcinogen and antibody is respected. Mucosal antibodies could decrease both systemic uptake and metabolic activation at the site of entry. Systemic antibodies may change the kinetics of carcinogen metabolism and redistribute carcinogens within the organism. Antibody-mediated redistribution may favor metabolism in less sensitive distal organs and thus result in lower concentrations of adduct-forming species at mucosal surfaces and other sensitive cells. This may be accomplished by avoiding threshold concentrations of carcinogens required for triggering carcinogen-mediated cytochrome P450 induction and tumor promotion. Studies at immunoprophylactic intervention in carcinogenicity are discussed. PMID- 15755637 TI - Synthesis and immunobiological activity of base substituted 2-amino-3-(purin-9 yl)propanoic acid derivatives. AB - 2-Amino-3-(purin-9-yl)propanoic acids substituted at position 6 of the purine base moiety by dimethylamino, cyclopropylamino, pyrrolidin-1-yl, hydroxy, and sulfanyl group as well as their 2-aminopurine analogues were prepared from corresponding 9-(2,2-diethoxyethyl)purines and 2-aminopurines, respectively, by the Strecker synthesis. 2-Aminopropanoic acid derivatives were tested for their immunostimulatory and immunomodulatory potency. Some of these compounds significantly enhanced secretion of chemokines RANTES and MIP-1alpha, the most potent was 2-amino-6-sulfanylpurine derivative. Most of these compounds also augmented NO biosynthesis triggered primarily by IFN-gamma. PMID- 15755638 TI - A comparison between two polarizability parameters in chemical--biological interactions. AB - The polarizability of a molecule, an important physical property, is currently attracting our attention particularly in the area of QSAR (quantitative structure activity relationships) for chemical-biological interactions. Our primary focus in the present study has been upon the computational aspects by using NVE (sum of the valence electrons) as a means for estimating polarizability, we have been surprised at its utility. In this report we demonstrate how NVE can be related to the calculated polarizability from a variety of efforts to better understand the subject. A comparison between the use of two polarizability parameters, that is, NVE and CMR (calculated molar refractivity) in the formulation of QSAR for chemical-biological interactions has been also discussed. PMID- 15755639 TI - Synthesis and evaluation of new omega-borono-alpha-amino acids as rat liver arginase inhibitors. AB - Recent studies have demonstrated that arginase plays important roles in pathologies such as asthma or erectile dysfunctions. We have synthesized new omega-borono-alpha-amino acids that are analogues of the previously known arginase inhibitors S-(2-boronoethyl)-l-cysteine (BEC) and 2-amino-6 boronohexanoic acid (ABH) and evaluated them as inhibitors of purified rat liver arginase (RLA). In addition to the distance between the B(OH)(2) and the alpha amino acid functions, the position of the sulfur atom in the side chain also appears as a key determinant for the interaction with the active site of RLA. Furthermore, substitution of the alkyl side chain of BEC by methyl groups and conformational restriction of ABH by incorporation of its side chain in a phenyl ring led to inactive compounds. These results suggest that subtle interactions govern the affinity of inhibitors for the active site of RLA. PMID- 15755640 TI - Inhibitory effects of flavonol glycosides from Cinnamomum osmophloeum on inflammatory mediators in LPS/IFN-gamma-activated murine macrophages. AB - Four kaempferol glycosides were isolated from the leaves of Cinnamomum osmophloeum Kaneh, a Taiwan endemic tree. These compounds namely, kaempferitrin (1), kaempferol 3-O-beta-D-glucopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-7-O alpha-L-rhamnopyranoside (2), kaempferol 3-O-beta-D-apiofuranosyl-(1-->2)-alpha-L arabinofuranosyl-7-O-alpha-L-rhamnopyranoside (3), and kaempferol 3-O-beta-D apiofuranosy-(1-->4)-alpha-L-rhamnopyranosyl-7-O-alpha-L-rhamnopyranoside (4). The structure of compound 2 was determined by spectroscopic analyses and acid hydrolysis. The isolates 1-4 were evaluated as inhibitors of some macrophage functions involved in the inflammatory process. These four compounds inhibited lipopolysaccharide (LPS) and interferon (IFN)-gamma-induced nitric oxide (NO), and cytokines [tumor necrosis factor (TNF)-alpha and interleukin (IL)-12] in a dose-dependent manner. The concentration of 50% inhibition (IC(50)) of NO by compounds 1, 3, 4 were 40, 15, 20microM, respectively. In parallel, these concentrations were approximately in a similar manner to that observed for TNF alpha and IL-12 production. However, compound 2 inhibited NO and cytokines production by 30% at 100microM concentration. On the other hand, compounds 3 and 4 showed no inhibitory effect on the production of NO from macrophages, when inducible NO synthase was already expressed by the stimulation with LPS and IFN gamma. Taken together, our results provide evidence that isolates of C. osmophloeum possess an anti-inflammatory potential which constitutes a previously unrecognized biological activity. PMID- 15755641 TI - Synthesis of novel DNA cross-linking antitumour agents based on polyazamacrocycles. AB - We are seeking to develop more effective alkylating agents as antitumour agents. In previous work conformationally restricted nitrogen mustards were synthesised containing piperidine or pyrrolidine rings. The free bases were designed to be bifunctional alkylating agents via aziridinium ion formation and the effects of varying the distances between the two alkylating sites were studied. Some efficient cross-linkers of naked DNA were prepared but few of these compounds exhibited significant cytotoxicity in human tumour cells in vitro. We have extended this work by making tri- and tetra-azamacrocyclic compounds containing two to four potential alkylating sites. Most of these compounds were powerful DNA alkylating agents and showed cytotoxicity (IC(50) values 6-100microM) comparable with chlorambucil (45microM) and melphalan (8.5microM). In particular the cyclen derivative 2a was more than 10(4) times more effective at cross-linking DNA (2a XL(50)<<10nM) than chlorambucil (XL(50) 100microM), and showed significant cytotoxicity in human tumour cells in vitro. PMID- 15755642 TI - Pyrazolopyridine antiherpetics: SAR of C2' and C7 amine substituents. AB - A novel series of potent pyrazolo[1,5-a]pyridine inhibitors of herpes simplex virus 1 replication have been identified. Several complimentary synthetic methods were developed to allow facile access to a diverse set of analogs from common late stage intermediates. Detailed examination of the amine substituents at the C2' position of the pyrimidine and C7 position of the core pyrazolopyridine is described. The antiviral data suggests that non-polar amines are preferred for optimal activity. Additionally, the 2' position has been shown to require an NH group to retain activity levels similar to that of the gold standard acyclovir. PMID- 15755644 TI - Stabilization of guanine quadruplex DNA by the binding of porphyrins with cationic side arms. AB - Many aromatic ligands, including tetra-(N-methyl-4-pyridyl)porphyrin (TMPyP4), have been reported to bind and stabilize quadruplex structure of telomeric DNA. We synthesized novel quadruplex-interacting porphyrins with cationic pyridinium and trimethylammonium arms at para- or meta-position of all phenyl groups of tetratolyl porphyrin. An antiparallel quadruplex structure was found to be stabilized more greatly by the meta-isomers than by the para-isomers and well studied TMPyP4, as revealed by the increase in melting temperature of the quadruplex. One mole equivalent of the isomers was sufficient to stabilize the quadruplex. From the results of absorption, induced circular dichroism, and fluorescence resonance energy transfer spectroscopic methods, the unique site for the porphyrin binding is suggested to be the external guanine tetrad or groove of the quadruplex. The cationic side arms played a key role in the stabilization of the quadruplex structure. PMID- 15755643 TI - Porphyrin substituted phosphoramidites: new building blocks for porphyrin oligonucleotide syntheses. AB - Thymidine phosphoramidites containing trispyridylphenyl and tetraphenylporphyrin chromophores attached via a short amide linker in the 3'-position have been synthesized and used as building blocks in solid-phase synthesis of self complementary 8-mer oligonucleotides 3'-T-5'-GCGCGCA-3' and 5'-ACGCGCGT-3'. To our knowledge, these are the first porphyrin-oligonucleotide conjugates carrying the porphyrin chromophores in the 3'-position. Chain assembly was achieved by automated solid-phase synthesis and by inexpensive straightforward 'in flask' modification of commercially available solid supported oligonucleotides. This approach allows the synthesis of modified oligonucleotides without using costly instrumentation for automated DNA synthesis. Porphyrin-containing self complementary oligonucleotides are expected to be a valuable model for drug binding studies and determination of conformational changes in DNA sequences using circular dichroism. PMID- 15755645 TI - Evaluation of macrocyclic Grb2 SH2 domain-binding peptide mimetics prepared by ring-closing metathesis of C-terminal allylglycines with an N-terminal beta-vinyl substituted phosphotyrosyl mimetic. AB - Preferential binding of ligands to Grb2 SH2 domains in beta-bend conformations has made peptide cyclization a logical means of effecting affinity enhancement. This is based on the concept that constraint of open-chain sequences to bend geometries may reduce entropy penalties of binding. The current study extends this approach by undertaking ring-closing metathesis (RCM) macrocyclization between i and i+3 residues through a process involving allylglycines and beta vinyl-functionalized residues. Ring closure in this fashion results in minimal macrocyclic tetrapeptide mimetics. The predominant effects of such macrocyclization on Grb2 SH2 domain binding affinity were increases in rates of association (from 7- to 16-fold) relative to an open-chain congener, while decreases in dissociation rates were less pronounced (approximately 2-fold). The significant increases in association rates were consistent with pre-ordering of solution conformations to near those required for binding. Data from NMR experiments and molecular modeling simulations were used to interpret the binding results. An understanding of the conformational consequences of such i to i+3 ring closure may facilitate its application to other systems where bend geometries are desired. PMID- 15755647 TI - Syntheses of quinolone hydrochloride enantiomers from synthons (R)- and (S)-2 methylpiperazine. AB - A series of R and S enantiomers of 7-(3-methylpiperazin-1-yl) quinolone derivatives were synthesized from (R)- and (S)-tert-butyl 2-methylpiperazine-1 carboxylate and tested for their antibacterial activities on 14 kinds of bacteria. Although no distinct difference in in vitro antibacterial activities was observed, 2-64-fold difference between R and S enantiomers was observed in approximately 52% of cases. PMID- 15755646 TI - Synthesis, structural identification, and ligand binding of tropane ring analogs of paroxetine and an unexpected aza-bicyclo[3.2.2]nonane rearrangement product. AB - The structural requirements for high affinity at the serotonin transporter (5 HTT) have been investigated through the preparation of rigid paroxetine analogs. Tropane-derived analogs (4a-i) of paroxetine (2) were designed and synthesized as potential inhibitors of serotonin reuptake based on the structural and biological similarity between the two compound classes. Overall, the affinity of tropane derived analogs at the 5-HTT was found to be at least an order of magnitude lower than that of paroxetine and ranged from 2-400nM. The reduced affinity at the 5 HTT may be attributed to the inability of the rigid tropane-derived analogs to adopt conformations favored by the 5-HTT. Within the series of tropane analogs, the 2beta,3beta- and 2beta,3alpha-isomers, 4a and 4d, were the most potent at the DAT and NET and are also significantly more potent than paroxetine (2) suggesting that their reduced conformational flexibility maximizes residence time in conformations favored by these transporters. Examination of the previously published preparation and structural assignment of 4a by additional NMR and X-ray crystallographic data has established that nucleophilic addition to the intermediate 2beta-methanesulfonyloxymethyl-3beta-(4-fluorophenyl)tropane unexpectedly provided the aza-bicyclo[3.2.2]nonane derivative 10a. PMID- 15755649 TI - New sesquiterpenic phytotoxins establish unprecedented relationship between different groups of blackleg fungal isolates. AB - A comprehensive search for sesquiterpenic metabolites produced by isolates of the blackleg fungus [Leptosphaeria maculans (Desm.)] Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.] revealed that an isolate pathogenic on both canola and brown mustard (IBCN 18) and two isolates pathogenic on brown mustard (Laird 2 and Mayfair 2) produced similar sesquiterpenes. The isolation, chemical structure elucidation, and phytotoxicity of these new sesquiterpenes with silphinene and selinene type skeletons is reported. This is the first time that an isolate virulent on canola and brown mustard is found to produce metabolites characteristic of both virulent (sirodesmins) and avirulent (phomalairdenones) L. maculans/P. lingam. In the context of grouping the various isolates of L. maculans/P. lingam, this work suggests an additional pathogenicity group comprising isolates that produce both sirodesmins and phomalairdenones and are virulent on both canola and brown mustard. PMID- 15755648 TI - Design, synthesis, and biological evaluation of N-acetyl-2 carboxybenzenesulfonamides: a novel class of cyclooxygenase-2 (COX-2) inhibitors. AB - N-Acetyl-2-carboxybenzenesulfonamide (11), and a group of analogues possessing an appropriately substituted-phenyl substituent (4-F, 2,4-F(2), 4-SO(2)Me, 4 OCHMe(2)) attached to its C-4, or C-5 position, were synthesized for evaluation as selective cyclooxygenase-2 (COX-2) inhibitors. In vitro COX-1/COX-2 inhibition studies showed that 11 is a more potent inhibitor (COX-1 IC(50)=0.06microM; COX-2 IC(50)=0.25microM) than aspirin (COX-1 IC(50)=0.35microM; COX-2 IC(50)=2.4microM), and like aspirin [COX-2 selectivity index (S.I.)=0.14], 11 is a nonselective COX-2 inhibitor (COX-2 S.I.=0.23). Regioisomers having a 2,4 difluorophenyl substituent attached to the C-4 (COX-2 IC(50)=0.087microM; COX-2 S.I. >1149), or C-5 (COX-2 IC(50)=0.77microM, SI>130), position of 11 exhibited the most potent and selective COX-2 inhibitory activity relative to the reference drug celecoxib (COX-1 IC(50)=33.1microM; COX-2 IC(50)=0.07microM; COX-2 S.I.=472). N-Acetyl-2-carboxybenzenesulfonamide (11, ED(50)=49 mg/kg), and its C 4 2,4-difluorophenyl derivative (ED(50)=91 mg/kg), exhibited superior antiinflammatory activity (oral dosing) in a carrageenan-induced rat paw edema assay compared to aspirin (ED(50)=129 mg/kg). These latter compounds exhibited comparable analgesic activity to the reference drug diflunisal, and superior analgesic activity compared to aspirin, in a 4% NaCl-induced abdominal constriction assay. A molecular modeling (docking) study indicated that the SO(2)NHCOCH(3) substituent present in N-acetyl-2-carboxy-4-(2,4 fluorophenyl)benzenesulfonamide, like the acetoxy substituent in aspirin, is suitably positioned to acetylate the Ser(530) hydroxyl group in the COX-2 primary binding site. The results of this study indicate that the SO(2)NHCOCH(3) pharmacophore present in N-acetyl-2-carboxybenzenesulfonamides is a suitable bioisostere for the acetoxy (OCOMe) group in aspirin. PMID- 15755650 TI - A topological substructural approach applied to the computational prediction of rodent carcinogenicity. AB - The carcinogenic activity has been investigated by using a topological substructural molecular design approach (TOPS-MODE). A discriminant model was developed to predict the carcinogenic and noncarcinogenic activity on a data set of 189 compounds. The percentage of correct classification was 76.32%. The predictive power of the model was validated by three test: an external test set (compounds not used in the develop of the model, with a 72.97% of good classification), a leave-group-out cross-validation procedure (4-fold full cross validation, removing 20% of compounds in each cycle, with a good prediction of 76.31%) and two external prediction sets (the first and second exercises of the National Toxicology Program). This methodology evidenced that the hydrophobicity increase the carcinogenic activity and the dipole moment of the molecule decrease it; suggesting the capacity of the TOPS-MODE descriptors to estimate this property for new drug candidates. Finally, the positive and negative fragment contributions to the carcinogenic activity were identified (structural alerts) and their potentialities in the lead generation process and in the design of 'safer' chemicals were evaluated. PMID- 15755651 TI - Fluorinated phenylcyclopropylamines. Part 4: effects of aryl substituents and stereochemistry on the inhibition of monoamine oxidases by 1-aryl-2-fluoro cyclopropylamines. AB - A series of para-ring-substituted (E)- and (Z)-1-aryl-2-fluorocyclopropylamines were examined as inhibitors of recombinant human liver monoamine oxidase A (MAO A) and B (MAO B). Unlike the parent 1-phenylcyclopropylamine, which is a selective inhibitor of MAO B, both (E)- and (Z)-diastereomers of derivatives having fluorine at the 2-position of the cyclopropane ring were potent and selective irreversible inhibitors of MAO A. Both electron releasing groups (Me, OMe) and electron attracting groups (Cl, F) substituted in the para-position caused a modest increase in activity. Geminal difluoro-substitution caused a loss of potency of 100-fold compared to either (E)- or (Z)-monofluorinated analogue. Surprisingly, (1S,2R)-2-fluoro-1-phenylcyclopropylamine and the (1R,2S) enantiomer were essential equally potent as inhibitors of MAO A and MAO B. None of the tested 1-aryl-2-fluorocyclopropylamines exhibited significant inhibition of tyramine oxidase. PMID- 15755652 TI - Benzotropolone inhibitors of estradiol methylation: kinetics and in silico modeling studies. AB - Natural and synthetic benzotropolone compounds were assessed in vitro for their ability to inhibit hydroxyestradiol methylation by catechol-O-methyltransferase (COMT). The compounds were also modeled in silico with a homology model of human COMT. Purpurogallin (1), purpurogallin carboxylic acid (2), and theaflavin-3,3' digallate (6) were the most potent inhibitors of 2-hydroxy and 4-hydroxyestradiol methylation (IC(50) 0.22-0.50microM). Compounds 1 and 6 decreased the V(max) and increased the K(m) of COMT, indicating a mixed-type inhibition. Compounds 1 and 2 bound to COMT by inserting the six-membered ring of the benzotropolone into the active site. Decreased acidity of the hydroxyl groups on this ring or increased bulkiness reduced potency. Compound 6 bound by inserting the galloyl ester into the active site, which allowed the compound to overcome increased bulkiness and resulted in restored potency. Further studies are needed to determine the impact in vivo of COMT inhibition by these compounds. PMID- 15755653 TI - Synthesis of potent and selective inhibitors of Candida albicans N myristoyltransferase based on the benzothiazole structure. AB - Two parallel synthetic methods using solid-supported reagents were established to examine the rapid optimization of weak hit compound 1. Several compounds showed high potency in the low nanomolar range against N-myristoyltransferase. The structure-activity relationship (SAR) and antifungal activities of a series of novel 2-aminobenzothiazole N-myristoyltransferase inhibitors are presented. PMID- 15755654 TI - Lysine-spermine conjugates: hydrophobic polyamine amides as potent lipopolysaccharide sequestrants. AB - Lipopolysaccharides (LPS), otherwise termed 'endotoxins', are outer-membrane constituents of Gram-negative bacteria. Lipopolysaccharides play a key role in the pathogenesis of 'Septic Shock', a major cause of mortality in the critically ill patient. Therapeutic options aimed at limiting downstream systemic inflammatory processes by targeting lipopolysaccharide do not exist at the present time. We have defined the pharmacophore necessary for small molecules to specifically bind and neutralize LPS and, using animal models of sepsis, have shown that the sequestration of circulatory LPS by small molecules is a therapeutically viable strategy. In this paper, the interactions of a focused library of lysine-spermine conjugates with lipopolysaccharide (LPS) have been characterized. Lysine-spermine conjugates with the epsilon-amino terminus of the lysinyl moiety derivatized with long-chain aliphatic hydrophobic substituents in acyl or alkyl linkage bind and neutralize bacterial lipopolysaccharides, and may be of use in the prevention or treatment of endotoxic shock states. PMID- 15755655 TI - Structural modification of phenylpropanoid-derived compounds and the effects on their participation in redox processes. AB - Oxidation and reduction processes are fundamental to many of the proposed mechanisms by which dietary phytochemicals are thought to exert protective effects against cardiovascular disease and some cancers. An understanding of the redox chemistry of these compounds is essential in assessing their potential to participate in these processes. Phenylpropanoid-derived compounds were selected and synthesised where required to represent many of the structural features found in this important group of compounds. Using electron paramagnetic resonance spectroscopy and computational chemistry a structure-redox activity relationship was obtained. Good correlation of computational and experimental results was observed for the mono-hydroxylated compounds. This demonstrated the value of computational chemistry in obtaining information about compounds, not readily available and the effect of electron delocalisation on parent radical stability. For compounds containing more than one hydroxyl, the relationship was found to be more complex. The importance of quinone formation in compounds containing more than one hydroxyl substituent was highlighted, as this was found to have a significant effect on stabilisation and therefore, their participation in redox processes. PMID- 15755656 TI - QSAR study for a novel series of ortho monosubstituted phenoxy analogues of alpha1-adrenoceptor antagonist WB4101. AB - A number of (S)- and (R)-2-[(2-phenoxyethyl)aminomethyl]-1,4-benzodioxanes unsubstituted or ortho monosubstituted at the phenoxy moiety were synthesized and tested in binding assays on the alpha(1a)-AR, alpha(1b)-AR, alpha(1d)-AR and the 5-HT(1A) receptor. The affinity values of the new compounds 1-16 were compared with those of the enantiomers of the 2,6-dimethoxyphenoxy analogue, the well known alpha(1) antagonist WB4101, finding that the unsubstituted derivative (S)-1 and the o-methyl, the o-t-butyl, the o-fluoro and the o-methoxy derivatives, (S) 2, (S)-4, (S)-8 and (S)-16, respectively, display a significantly specific 5 HT(1A) affinity, very close, with the exception of (S)-4, to the almost nanomolar one of (S)-WB4101. Otherwise, sensible affinity decreases were recorded for the three alpha(1)-AR subtypes. A classical quantitative structure-activity relationship (Hansch) analysis was successfully applied to compounds (S)-1 to (S) 16 and (S)-WB4101 to rationalize such binding data. PMID- 15755657 TI - Anti-inflammatory activity and QSAR studies of compounds isolated from Hyacinthaceae species and Tachiadenus longiflorus Griseb. (Gentianaceae). AB - Twenty-two homoisoflavanones and structurally related compounds isolated from plants were screened for anti-inflammatory activity. Seventeen compounds were isolated from southern African Hyacinthaceae species, one from the Madagascan gentian Tachiadenus longiflorus Griseb. and four were of synthetic origin. Inhibition of prostaglandin synthesis in cell microsomal fractions was first evaluated, followed by screening for specific inhibition of isolated cyclooxygenase enzymes (COX-1 and COX-2). Six homoisoflavanone and structurally related compounds showed significantly high levels of anti-inflammatory activity in the microsomal fraction assay. Only one compound exhibited a high level of anti-inflammatory activity in the COX-1 enzyme assay and no significant activity was detected in the COX-2 enzyme assay. Biological screening was followed by a computer-based quantitative structure-activity relationship (QSAR) study. The physicochemical descriptors: strain energy, heat of formation, volume, surface area, aqueous phase energy, dipole moment, enthalpy, entropy, molar refractivity, parachor, density, refractive index, surface tension, polarizability, logP, Van der Waals interaction energy, Coulombic interaction energy and nonbonded interaction energy were used to characterize the structures of the homoisoflavanones and structurally related compounds. This study produced three equations with significant prediction values for the anti-inflammatory activity of the compounds investigated. The derived models also provided valuable parameter guidelines for those properties influencing the anti-inflammatory activity of the studied compounds. PMID- 15755658 TI - Methyltrioxorhenium: a new catalyst for the activation of hydrogen peroxide to the oxidation of lignin and lignin model compounds. AB - The oxidative degradation of lignin under totally chlorine free conditions is one of the most relevant targets for the design of environmental friendly pulping and bleaching industrial processes. Methyltrioxorhenium was found a powerful and promising catalyst for the oxidation of both phenolic and non-phenolic lignin model compounds by use of hydrogen peroxide as primary oxidant. Three different technical lignins, hydrolytic sugar cane lignin (SCL), red spruce kraft lignin (RSL) and a hardwood organosolvent lignin (OSL), that are representative examples of widely diffused para-hydroxyphenyl-guaiacyl, guaiacyl and guaiacyl-syringyl lignins, were also extensively degraded under similar experimental conditions. PMID- 15755659 TI - Bridge-linked bis-quaternary ammonium anti-microbial agents: relationship between cytotoxicity and anti-bacterial activity of 5,5'-[2,2' (tetramethylenedicarbonyldioxy)-diethyl]bis(3-alkyl-4-methylthiazonium iodide)s. AB - We examined the correlation between the anti-bacterial activity against Escherichia coli and the cytotoxicity of five synthesized bridge types of bis quaternary ammonium compounds (bis-QACs) as follows: thioether type, 4,4'-(p xylydithio)bis(1-octylpyridinium iodide) (4DTBP-X,8); amide type, N,N' tetramethylenebis(1-dodecyl-4-carbamoylpyridinium iodide) (4BCAP-4,12), N,N' (phenylene)bis(1-octyl-4-carbamoylpyridinium iodide) (4BCAP-P,8); anti-amide type, 4,4'-(tetramethylenedicarbonyldiamino)bis(1-octylpyridinium iodide) (4DCABP 4,8), 4,4'-(phenylenedicarbonyldiamino)bis(1-octylpyridinium iodide) (4DCABP P,8); ester type, 4,4'-(1,6-hexamethylenedioxydicarbonyl)bis(1-dodecylpyridinium iodide) (4DOCBP-6,12); and an anti-ester type, 5,5'-[2,2' (tetramethylenedicarbonyldioxy)diethyl]bis(3-alkyl-4-methylthiazolium iodide) (5DEBT-4,n, The letter n indicates the carbon number of the alkyl group). 5DEBT 4,8 showed low cytotoxicity (LD(50)) to human erythrocytes (97+/-6microM) and the NB1RGB cell line (111+/-20microM) and remarkable anti-bacterial activity (MIC) toward E. coli K12 W3110 (7.9microM). Moreover, 5DEBT-4,8 indicated 1144 conformers by global minimum analysis and had two minimum dGW (solvation free energy) points as well as 4DTBP-6,8, which had been previously examined and concluded to be a significant useful anti-bacterial compound. PMID- 15755660 TI - Inhibitory activity of a ceramide library on interleukin-4 production from activated T cells. AB - Allergic diseases are hypersensitivity disorders associated with the production of specific immunoglobulin E (IgE) to environmental allergens. Interleukin (IL) 4, produced primarily by CD4(+) T cells, is an important stimulus for the switch of the antibody isotype to IgE in both mice and humans. In this study we investigated the inhibitory activity of IL-4 production in activated T cells by screening ceramide derivatives prepared by solid phase combinatorial chemistry. Many ceramide derivatives significantly inhibited IL-4 production in T cells. In particular, ceramide derivatives with a lauroyl group showed strong inhibitory activities on IL-4 production in both phorbol 12-myristate 13-acetate (PMA) activated EL4 T cells and antigen-primed cells, suggesting that they can be used as compounds for the development of anti-allergic agents. PMID- 15755661 TI - Synthesis and biological evaluation of branched and conformationally restricted analogs of the anticancer compounds 3'-C-ethynyluridine (EUrd) and 3'-C ethynylcytidine (ECyd). AB - The synthesis of branched and conformationally restricted analogs of the anticancer nucleosides 3'-C-ethynyluridine (EUrd) and 3'-C-ethynylcytidine (ECyd) is presented. Molecular modeling and (1)H NMR coupling constant analysis revealed that the furanose rings of all analogs except the LNA analog are conformationally biased towards South conformation, and are thus mimicking the structure of ECyd. All target nucleosides were devoid of anti-HIV or anticancer activity. PMID- 15755662 TI - Synthesis and evaluation of the permeability transition inhibitory characteristics of paramagnetic and diamagnetic amiodarone derivatives. AB - Several amiodarone analogues were synthesized varying the 2-substituent on the benzofuran ring and diethylaminoethyl side chain of phenolether by introducing 2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole and 1,2,5,6-tetrahydropyridine nitroxides or their amino or hydroxylamino precursors. The new compounds were screened on isolated mitochondria and perfused heart and their toxicity was evaluated on WRL-68 liver cells and H9C2 cardiomyocytes. Most of the newly synthesized derivatives exerted uncoupling effect on the mitochondrial oxidative phosphorilation at higher concentrations, compared to amiodarone and one of the modified amiodarone analogues showed an effect similar to that of amiodarone on the mitochondrial permeability transition and on restoring of mitochondrial high energy phosphate metabolites in perfused hearts. This amiodarone analogue can be new leading compound among the experimental amiodarone analogues with the same or enhanced efficiency of amiodarone, but with less side effects. PMID- 15755663 TI - Design, synthesis and evaluation of 2,4-diaminoquinazolines as inhibitors of trypanosomal and leishmanial dihydrofolate reductase. AB - This paper describes the design, synthesis and evaluation of a series of 2,4 diaminoquinazolines as inhibitors of leishmanial and trypanosomal dihydrofolate reductase. Compounds were designed by a generating virtual library of compounds and docking them into the enzyme active site. Following their synthesis, they were found to be potent and selective inhibitors of leishmanial dihydrofolate reductase. The compounds were also found to have potent activity against Trypanosoma brucei rhodesiense, a causative organism of African trypanosomiasis and also against Trypanosoma cruzi, the causative organism of Chagas disease. There was significantly lower activity against Leishmania donovani, one of the causative organisms of leishmaniasis. PMID- 15755664 TI - Enzymatic hydrolysis of stampidine and other stavudine phosphoramidates in the presence of mammalian proteases. AB - Mammalian proteases have not been implicated in the metabolism of any nucleoside phosphoramidate prodrug. The results presented herein provide unprecedented and conclusive experimental evidence that mammalian proteases are capable of hydrolyzing stavudine phosphoramidates. Specifically, cathepsin B and Proteinase K are able to metabolize stampidine and other phosphoramidate derivatives of stavudine. Additionally, cathepsin B exhibits chiral selectivity at the phosphorus center. The elucidation of the metabolic pathways leading to activation of stampidine may provide the basis for pharmacologic interventions aimed at modulating the metabolism and thereby improving the therapeutic window of stampidine as an anti-HIV agent. PMID- 15755665 TI - Discovery of novel non-peptidic ketopiperazine-based renin inhibitors. AB - Ketopiperazine 2 was designed from a previously published analog. Compound 2 was shown to be a novel, potent inhibitor of renin that, when administered orally, lowered blood pressure in a hypertensive double transgenic (human renin and angiotensinogen) mouse model. Compound 2 was further optimized to sub-nanomolar potency by designing an analog that addressed the S3 sub-pocket of the renin enzyme (16). PMID- 15755666 TI - Synthesis and inhibitory activity of benzoic acid and pyridine derivatives on influenza neuraminidase. AB - Based upon the activity and X-ray crystallographic studies of tri-substituted benzene derivatives containing carboxylic acid, acetamido and guanidine groups, we investigated the effect of the fourth substituent to fulfill the fourth pocket of neuraminidase enzyme. The groups selected as fourth substituents were hydroxymethyl, hydroxyethyl, oxime and amino. These tetra-substituted benzene derivatives were synthesized and evaluated for neuraminidase inhibitory activity. All these compounds were found to have poorer IC(50) values than the tri substituted compounds. Further, benzene ring was replaced by pyridine ring and di, tri and tetra-substituted pyridine derivatives were synthesized. The activity of the pyridine derivatives was comparable to benzene derivatives. The fourth substituent seems to disturb the binding of the other three substituents, so the activity is reduced as compared to tri-substituted benzene and pyridine derivatives. PMID- 15755667 TI - The study on the interaction between seryl-histidine dipeptide and proteins by circular dichroism and molecular modeling. AB - The selective cleavage of proteins is very important in key biological processes. Chemical (nonenzymatic) reagents such as cyanogen bromide and transition metal complexes are used extensively with great defects. In this paper, the binding of seryl-histidine dipeptide (abbreviated as SH) with bovine serum albumin (BSA) and lysozyme were investigated by the circular dichroism spectroscopy (CD) at 298K, molecular docking studies and quantum chemical calculations based on the previous results of polyacrylamide gel electrophoresis (PAGE). From the studies of CD, it showed that SH interacted strongly with BSA and lysozyme. The change percentages of the secondary structures of BSA and lysozyme were calculated. The contents of the beta-sheets decreased remarkably. It indicated that the interactions between SH and proteins could break the hydrogen bonds of beta-sheets selectively. The docking studies between SH and BSA showed that the position of the oxygen atom of the hydroxyl group of SH (O(12)) was in favor of a nucleophilic attack on carbon atom of the amide bond of a beta-sheet (C(34)) because the distance between O(12) and C(34) was 3.37A. Natural charges, natural atomic hybrid percentages and square sums of HOMO coefficients calculated by the NBO and population analysis at HF/6-31G* supported the suggested mechanism. And so SH may be an interesting agent for the therapeutic use. PMID- 15755668 TI - The extent of damage following repeated injury to cultured hippocampal cells is dependent on the severity of insult and inter-injury interval. AB - Recent evidence suggests repeated mild brain trauma may result in cumulative damage. We investigated cell damage and death in hippocampal cultures following repeated mechanical trauma in vitro by measuring propidium iodide (PrI) uptake, release of neuron-specific enolase (NSE) and glial S-100beta protein, and performing neuronal counts. Cultures receiving two mild injuries (31% stretch) 1 or 24 h apart displayed different profiles of PrI uptake and S-100beta release, although neuronal loss and NSE release was similar in both paradigms. Cells receiving a subthreshold, low-level stretch (10%) repeated several times eventually stained with PrI. Cultures administered 10% stretch before mild injury released less S-100beta than mild injury alone, suggesting a preconditioning effect. Lastly, exogenous S-100beta applied to injured cultures decreased PrI uptake, implying a protective role. These results suggest cumulative damage is dependent on injury severity and inter-injury interval, and that neurons and glia react differently to various injury paradigms. PMID- 15755669 TI - Increased oxidative protein and DNA damage but decreased stress response in the aged brain following experimental stroke. AB - Aged individuals experience the highest rate of stroke and have less functional recovery, but do not have larger infarcts. We hypothesized that aged individuals experience greater sublethal damage in peri-infarct cortex. Focal cortical stroke was produced in aged and young adult animals. After 30 min, 1, 3 and 5 days brain sections and Western blot were used to analyze markers of apoptotic cell death, oxidative DNA and protein damage, heat shock protein (HSP) 70 induction, total neuronal number and infarct size. Focal stroke produces significantly more oxidative DNA and protein damage and fewer cells with HSP70 induction in peri infarct cortex of aged animals. There is no difference in infarct size or the number of cells undergoing apoptosis between aged and young adults. Stroke in the aged brain is associated with a greater degree of DNA and protein damage and a reduced stress response in intact, surviving tissue that surrounds the infarct. PMID- 15755670 TI - Triptolide protects dopaminergic neurons from inflammation-mediated damage induced by lipopolysaccharide intranigral injection. AB - Converging lines of evidence suggest that neuroinflammatory processes may account for the progressive death of dopaminergic neurons in Parkinson's disease (PD). Therefore, anti-inflammatory strategies have attracted much interest for their potential to prevent further deterioration of PD. Our previous study showed that triptolide, a traditional Chinese herbal compound with anti-inflammatory and immunosuppressive properties, protected dopaminergic neurons from lipopolysaccharide (LPS)-induced damage in primary embryonic midbrain cell cultures. To examine further if triptolide can protect dopaminergic neurons from inflammation-mediated damage in vivo, microglial activation and injury of dopaminergic neurons were induced by LPS intranigral injection, and the effects of triptolide treatment on microglial activation and survival ratio and function of dopaminergic neurons were investigated. Our results demonstrated that microglial activation induced by a single intranigral dose of 10 mug of LPS reduced the survival ratio of tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the substantia nigra pars compacta (SNpc) to 29% and the content of dopamine (DA) in striatum to 37% of the non-injected side. Intriguingly, treatment with triptolide of 5 mug/kg for 24 days once per day dramatically improved the survival rate of TH-ir neurons in the SNpc to 79% of the non-injected side. Meanwhile, treatment with triptolide of 1 or 5 mug/kg for 24 days once per day significantly improved DA level in striatum to 70% and 68% of the non-injected side, respectively. Complement receptor 3 (CR3) immunohistochemical staining revealed that triptolide treatment potently inhibited LPS-elicited deleterious activation of microglia in SNpc. The excessive production of cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, was significantly abolished by triptolide administration. These results, together with our previous data in vitro, highly suggest the effectiveness of triptolide in protecting dopaminergic neurons against inflammatory challenge. PMID- 15755671 TI - Caspase-3-mediated cleavage of PHF-1 tau during apoptosis irrespective of excitotoxicity and oxidative stress: an implication to Alzheimer's disease. AB - Excitotoxicity, oxidative stress, and apoptosis have been recognized as routes to neuronal death in various neurological diseases. We examined the possibility that PHF-1 tau, a substrate for various proteases, would be selectively cleaved depending upon routes of neuronal death. Cleavage form of PHF-1 tau was not observed in cortical cell cultures exposed to excitotoxins or oxidative stress that cause neuronal cell necrosis. PHF-1 tau was cleaved within 8 h following exposure of cortical cell cultures to apoptosis-inducing agents. This cleavage was blocked by inclusion of zDEVD-fmk, an inhibitor of caspase-3, and accompanied by activation of caspase-3. Levels and cleavage of PHF-1 tau were markedly increased in AD brain compared with control. Moreover, PHF-1 tau and active caspase-3 were colocalized mostly in tangle-bearing neurons. The current findings suggest that PHF-1 tau is cleaved by caspase-3 during apoptosis and neurodegenerative process in AD. PMID- 15755672 TI - Differential effects of oligomeric and fibrillar amyloid-beta 1-42 on astrocyte mediated inflammation. AB - Activated glia, as a result of chronic inflammation, are associated with amyloid beta peptide (Abeta) deposits in the brain of Alzheimer's disease (AD) patients. In vitro, glia are activated by Abeta inducing secretion of pro-inflammatory molecules. Recent studies have focused on soluble oligomers (or protofibrils) of Abeta as the toxic species in AD. In the present study, using rat astrocyte cultures, oligomeric Abeta induced initial high levels of IL-1beta decreasing over time and, in contrast, fibrillar Abeta increased IL-1beta levels over time. In addition, oligomeric Abeta, but not fibrillar Abeta, induced high levels of iNOS, NO, and TNF-alpha. Our results suggest that oligomers induced a profound, early inflammatory response, whereas fibrillar Abeta showed less increase of pro inflammatory molecules, consistent with a more chronic form of inflammation. PMID- 15755674 TI - Glutamate transporter function of rat hippocampal astrocytes is impaired following the global ischemia. AB - Astroglial glutamate transporters, GLT-1 and GLAST, play an essential role in removing released glutamate from the extracellular space and are essential for maintaining a low concentration of extracellular glutamate in the brain. It was hypothesized that impaired function of glial glutamate transporters induced by transient global ischemia may lead to an elevated level of extracellular glutamate and subsequent excitotoxic neuronal death. To test this hypothesis, in the present study, we performed whole-cell patch-clamp recording of hippocampal CA1 astrocytes in control or postischemic slices, and measured glutamate transporter activity by recording glutamate-evoked transporter currents. Six to 24 h after global ischemia, maximal amplitude of glutamate transporter currents recorded from postischemic CA1 astrocytes was significantly reduced. Western blotting analysis indicated that transient global ischemia decreased the protein level of GLT-1 in the hippocampal CA1 area without affecting GLAST protein level. Further real-time quantitative RT-PCR assays showed that global ischemia resulted in a decrease in GLT-1 mRNA level of hippocampal CA1 region. Global ischemia induced reduction in GLT-1 expression and glutamate transporter function of CA1 astrocytes precedes the initiation of delayed neuronal death in CA1 pyramidal layer. The present study provides the evidence that transient global ischemia downregulates glutamate transporter function of hippocampal CA1 astrocytes by decreasing mRNA and protein levels of GLT-1. PMID- 15755673 TI - Repetitive febrile seizures in rat pups cause long-lasting deficits in synaptic plasticity and NR2A tyrosine phosphorylation. AB - Adult rats with early-life frequently repetitive febrile seizures (FRFS), but not single febrile seizure (SFS), exhibited impaired performance in inhibitory avoidance tasks but without significant hippocampal neuronal loss. The mechanisms of long-term memory impairment in the hippocampus of adult rats with early-life FRFS remain unknown. Using a heated-air febrile seizures (FS) paradigm, male rat pups were subjected to single or nine episodes of brief FS at days 10 to 12 postpartum. We found that early-life FRFS led to long-term bidirectional modulation in hippocampal synaptic plasticity, i.e., impaired long-term potentiation and facilitated long-term depression. Three hours after inhibitory avoidance training, phosphorylation of hippocampal extracellular signal-regulated kinase (ERK) 1/2 was significantly less in the FRFS group than in controls. Furthermore, there was a selective alteration in NMDA receptor-mediated ERK1/2 phosphorylation in the hippocampus of the FRFS group. Although the expression levels of NMDA receptor subunits and interaction of NMDA receptor and postsynaptic density 95 did not alter quantitatively, there was a specific alteration in NR2A, but not NR2B, subunit tyrosine phosphorylation after NMDA stimulation in the FRFS group. These data offer a potential molecular explanation for the hippocampus-dependent memory deficits observed in the rats with early life FRFS. PMID- 15755675 TI - Heterogeneous GABAA receptor subunit expression in pediatric epilepsy patients. AB - The gamma-amino-butyric acid type A receptors (GABAAR) are a heteropentameric receptor complex, composed of 16 possible subunits in various combinations, forming a ligand-gated ion channel. Subunit composition is the primary determinant of GABAAR physiology and pharmacology. Here we have measured mRNA levels for 16 GABAAR subunits in isolated dentate granule neurons (DGN) from eight pediatric patients undergoing resective surgery for intractable epilepsy. We found tightly correlated expression of a subset of GABAAR subunit mRNAs within a single DGN (alpha1, gamma1, and gamma2; alpha4, alpha5, and beta2; alpha4 and beta3). Analysis of inter-patient variability (ANOVA) of eleven highly expressed GABAAR subunit mRNAs found seven of the subunits varied between patients, as did whole cell GABAAR currents. Due to inter-patient differences, there is heterogeneity in DGN GABAAR subunit mRNA and physiology within pediatric epilepsy patients. Patient-specific GABAAR expression might contribute to variability in anti-epileptic drug efficacy, side-effect profiles, and seizure susceptibility. PMID- 15755676 TI - Mitochondrial associated metabolic proteins are selectively oxidized in A30P alpha-synuclein transgenic mice--a model of familial Parkinson's disease. AB - Parkinson's disease (PD) is the most common neurodegenerative movement disorder and is characterized by the loss of dopaminergic neurons in the substantia nigra compacta. alpha-Synuclein is strongly implicated in the pathophysiology of PD because aggregated alpha-synuclein accumulates in the brains of subjects with PD, mutations in alpha-synuclein cause familial PD, and overexpressing mutant human alpha-synuclein (A30P or A53T) causes degenerative disease in mice or drosophila. The pathophysiology of PD is poorly understood, but increasing evidence implicates mitochondrial dysfunction and oxidative stress. To understand how mutations in alpha-synuclein contribute to the pathophysiology of PD, we undertook a proteomic analysis of transgenic mice overexpressing A30P alpha synuclein to investigate which proteins are oxidized. We observed more than twofold selective increases in specific carbonyl levels of three metabolic proteins in brains of symptomatic A30P alpha-synuclein mice: carbonic anhydrase 2 (Car2), alpha-enolase (Eno1), and lactate dehydrogenase 2 (Ldh2). Analysis of the activities of these proteins demonstrates decreased functions of these oxidatively modified proteins in brains from the A30P compared to control mice. Our findings suggest that proteins associated with impaired energy metabolism and mitochondria are particularly prone to oxidative stress associated with A30P mutant alpha-synuclein. PMID- 15755677 TI - Pentapeptides derived from Abeta 1-42 protect neurons from the modulatory effect of Abeta fibrils--an in vitro and in vivo electrophysiological study. AB - Short fragments and fragment analogues of beta-amyloid 1-42 peptide (Abeta1-42) display a protective effect against Abeta-mediated neurotoxicity. After consideration of our earlier results with in vitro bioassay of synthetic Abeta recognition peptides and toxic fibrillar amyloids, five pentapeptides were selected as putative neuroprotective agents: Phe-Arg-His-Asp-Ser amide (Abeta4-8) and Gly-Arg-His-Asp-Ser amide (an analogue of Abeta4-8), Leu-Pro-Tyr-Phe-Asp amide (an analogue of Abeta17-21), Arg-Ile-Ile-Gly-Leu amide (an analogue of Abeta30-34), and Arg-Val-Val-Ile-Ala amide (an analogue of Abeta38-42). In vitro electrophysiological experiments on rat brain slices demonstrated that four of these peptides counteracted with the field excitatory postsynaptic potential attenuating effect of Abeta1-42; only Arg-Val-Val-Ile-Ala amide proved inactive. In in vivo experiments using extracellular single-unit recordings combined with iontophoresis, all these pentapeptides except Arg-Val-Val-Ile-Ala amide protected neurons from the NMDA response-enhancing effect of Abeta1-42 in the hippocampal CA1 region. These results suggest that Abeta recognition sequences may serve as leads for the design of novel neuroprotective compounds. PMID- 15755678 TI - Accumulation of human SOD1 and ubiquitinated deposits in the spinal cord of SOD1G93A mice during motor neuron disease progression correlates with a decrease of proteasome. AB - Mutations in SOD1 cause selective motor neuron degeneration in familial amyotrophic lateral sclerosis patients and transgenic mice overexpressing the mutant enzyme. Formation and accumulation of ubiquitinated aggregates in motor neurons are thought to be involved in the toxic gain of function of mutant SOD1. The present study shows that the accumulation of soluble and detergent-insoluble mutant SOD1 in spinal cord of symptomatic SOD1G93A transgenic mice is due to impaired degradation of mutant SOD1 rather than to increased transcript levels. This effect was accompanied by a decrease of constitutive proteasome levels and a concomitant increase of immunoproteasome in the spinal cord homogenate which resulted in overall unchanged proteasome activity. A decrease of constitutive proteasome occurred in the motor neurons of SOD1G93A mice at the presymptomatic stage and became remarkable with the progression of the disease. This provides further evidence for an involvement of proteasome impairment in the toxicity of mutant SOD1. PMID- 15755679 TI - D3 dopamine receptors do not regulate neurogenesis in the subventricular zone of adult mice. AB - Testing the effects of drugs that stimulate endogenous neurogenesis in different species is important for the development of neural repair strategies in humans. We have previously shown in adult rats that a 14-day intracerebroventricular infusion of the D3 preferential agonist 7-hydroxydipropyl-amino-tetraline (7-OH DPAT) increases BrdU labeling of neural precursors in the subventricular zone of the anterior lateral ventricle (SVZ). Here, we show that such a treatment failed to affect neurogenesis in C57Bl/6 and FVB mice, even at a high dose or when infused into the neostriatum. We confirmed that such a treatment was effective in adult rats. Moreover, D3 receptor inhibition or genetic knockout failed to affect the neurogenesis in mice. These results raise the possibilities that neurogenesis is not regulated by D3 receptors in all species and, therefore, that D3 agonists like pramipexole may not be useful to harness endogenous neurogenesis in cell replacement strategies for Parkinson's disease. PMID- 15755680 TI - Transgenic expression of an expanded (GCG)13 repeat PABPN1 leads to weakness and coordination defects in mice. AB - Oculopharyngeal muscular dystrophy (OPMD) is a late-onset disorder caused by a (GCG)n trinucleotide repeat expansion in the poly(A) binding protein nuclear-1 (PABPN1) gene, which in turn leads to an expanded polyalanine tract in the protein. We generated transgenic mice expressing either the wild type or the expanded form of human PABPN1, and transgenic animals with the expanded form showed clear signs of abnormal limb clasping, muscle weakness, coordination deficits, and peripheral nerves alterations. Analysis of mitotic and postmitotic tissues in those transgenic animals revealed ubiquitinated PABPN1-positive intranuclear inclusions (INIs) in neuronal cells. This latter observation led us to test and confirm the presence of similar INIs in postmortem brain sections from an OPMD patient. Our results indicate that expanded PABPN1, presumably via the toxic effects of its polyalanine tract, can lead to inclusion formation and neurodegeneration in both the mouse and the human. PMID- 15755681 TI - Microarray analysis identifies an aberrant expression of apoptosis and DNA damage regulatory genes in multiple sclerosis. AB - To clarify the molecular mechanisms underlying multiple sclerosis (MS)-promoting autoimmune process, we have investigated a comprehensive gene expression profile of T cell and non-T cell fractions of peripheral blood mononuclear cells (PBMC) isolated from 72 MS patients and 22 age- and sex-matched healthy control (CN) subjects by using a cDNA microarray. Among 1258 genes examined, 173 genes in T cells and 50 genes in non-T cells were expressed differentially between MS and CN groups. Downregulated genes greatly outnumbered upregulated genes in MS. More than 80% of the top 30 most significant genes were categorized into apoptosis signaling-related genes of both proapoptotic and antiapoptotic classes. They included upregulation in MS of orphan nuclear receptor Nurr1 (NR4A2), receptor interacting serine/threonine kinase 2 (RIPK2), and silencer of death domains (SODD), and downregulation in MS of TNF-related apoptosis-inducing ligand (TRAIL), B-cell CLL/lymphoma 2 (BCL2), and death-associated protein 6 (DAXX). Furthermore, a set of the genes involved in DNA repair, replication, and chromatin remodeling was downregulated in MS. These results suggest that MS lymphocytes show a complex pattern of gene regulation that represents a counterbalance between promoting and preventing apoptosis and DNA damage of lymphocytes. PMID- 15755682 TI - PABPN1 overexpression leads to upregulation of genes encoding nuclear proteins that are sequestered in oculopharyngeal muscular dystrophy nuclear inclusions. AB - Oculopharyngeal muscular dystrophy (OPMD) is an adult-onset disease caused by expanded (GCN)12-17 stretches encoding the N-terminal polyalanine domain of the poly(A) binding protein nuclear 1 (PABPN1). OPMD is characterized by intranuclear inclusions (INIs) in skeletal muscle fibers, which contain PABPN1, molecular chaperones, ubiquitin, proteasome subunits, and poly(A)-mRNA. We describe an adenoviral model of PABPN1 expression that produces INIs in most cells. Microarray analysis revealed that PABPN1 overexpression reproducibly changed the expression of 202 genes. Sixty percent of upregulated genes encode nuclear proteins, including many RNA and DNA binding proteins. Immunofluorescence microscopy revealed that all tested nuclear proteins encoded by eight upregulated genes colocalize with PABPN1 within the INIs: CUGBP1, SFRS3, FKBP1A, HMG2, HNRPA1, PRC1, S100P, and HSP70. In addition, CUGBP1, SFRS3, and FKBP1A were also found in OPMD muscle INIs. This study demonstrates that a large number of nuclear proteins are sequestered in OPMD INIs, which may compromise cellular function. PMID- 15755683 TI - Developmental changes induced by graded prenatal systemic hypoxic-ischemic insults in rats. AB - In infants, a common consequence of systemic perinatal insults is disruption of neonatal brain development. Such insults can cause cerebral palsy, cognitive delay, epilepsy and other chronic neurologic deficits in children. The mechanisms underlying disruption of brain development after perinatal insults are poorly defined. To mimic human systemic insults, a transient prenatal hypoxic-ischemic insult model was developed in rodents. Ischemic animals showed reproducible histological lesions including oligodendrocyte loss, gliosis, and axonal disruption. Ischemic animals displayed persistent postnatal loss of oligodendrocyte lineage cells and cortical neurons, decreased cell proliferation, increased cell death, elevated pro-inflammatory cytokine levels, and impaired motor skills as young adults. Progressive ischemic intervals produced a graded pattern of injury. This systemic rodent prenatal hypoxic-ischemic insult accurately models human perinatal brain injury in several important criteria, including functional association of altered brain development with motor delay, and consequently provides novel insights into the pathogenesis of human perinatal brain insults. PMID- 15755684 TI - Expression time course and spatial distribution of activated caspase-3 after experimental status epilepticus: contribution of delayed neuronal cell death to seizure-induced neuronal injury. AB - Pilocarpine-induced status epilepticus (PCSE) is a widely used model to study neurodegeneration in limbic structures after prolonged epileptic seizures. However, mechanisms mediating neuronal cell death in this model require further characterization. Examining the expression time course and spatial distribution of activated caspase-3, we sought to determine the role of apoptosis in PCSE mediated neuronal cell death. Expression of activated caspase-3, predominantly located in neurons, was detected 24 h (amygdala; piriform and temporal cortex) and 7 days (hippocampus; amygdala; piriform, temporal and parietal cortex; thalamus) after PCSE with strongest induction being observed in the amygdala, the piriform cortex, and the hippocampus. Further analysis revealed TUNEL positivity (24 h and 7 days after SE) and a significant, progressive neuronal cell loss in all brain regions displaying caspase-3 activation. Corresponding to high levels of activated caspase-3 expression, neuronal cell loss was most pronounced in the amygdala, piriform cortex, and dorsal CA-1 hippocampus. These results demonstrate that apoptosis contributes significantly to PCSE-induced neuronal cell death. PMID- 15755685 TI - Cannabinoid CB1 antagonists possess antiparkinsonian efficacy only in rats with very severe nigral lesion in experimental parkinsonism. AB - We have observed that systemic administration of cannabinoid CB1 antagonists exerts antiparkinsonian effects in rats with very severe nigral lesion (>95% cell loss), but not in rats with less severe lesion (85-95% cell loss). Local injections into denervated striatum and corresponding globus pallidus reduced parkinsonian asymmetry. Infusions into lesioned substantia nigra enhanced motor asymmetries, but this effect was absent after very severe nigral lesion. At the striatal level, CB1 antagonists act enhancing dopamine D1 receptor function and reducing D2 receptor function. Striatal dopaminergic denervation did not affect cannabinoid CB1 receptor coupling to G proteins. These results suggest that (i) systemic administration of CB1 antagonists in rats with severe nigral degeneration is ineffective because striatopallidal-mediated motor effects are antagonized by nigra-mediated activity, and (ii) CB1 antagonists exert antiparkinsonian effects after very severe nigral degeneration because nigra mediated inhibition disappears. CB1 receptor antagonists that lack psychoactive effects might be of therapeutic value in the control of very advanced stage of Parkinson's disease in humans. PMID- 15755686 TI - Episodic-like memory deficits in the APPswe/PS1dE9 mouse model of Alzheimer's disease: relationships to beta-amyloid deposition and neurotransmitter abnormalities. AB - Transgenic mice made by crossing animals expressing mutant amyloid precursor protein (APPswe) to mutant presenilin 1 (PS1dE9) allow for incremental increases in Abeta42 production and provide a model of Alzheimer-type amyloidosis. Here, we examine cognition in 6- and 18-month old transgenic mice expressing APPswe and PS1dE9, alone and in combination. Spatial reference memory was assessed in a standard Morris Water Maze task followed by assessment of episodic-like memory in Repeated Reversal and Radial Water maze tasks. We then used factor analysis to relate changes in performance in these tasks with cholinergic markers, somatostatin levels, and amyloid burden. At 6 months of age, APPswe/PS1dE9 double transgenic mice showed visible plaque deposition; however, all genotypes, including double-transgenic mice, were indistinguishable from nontransgenic animals in all cognitive measures. In the 18-month-old cohorts, amyloid burdens were much higher in APPswe/PS1dE9 mice with statistically significant but mild decreases in cholinergic markers (cortex and hippocampus) and somatostatin levels (cortex). APPswe/PS1dE9 mice performed all cognitive tasks less well than mice from all other genotypes. Factor and correlation analyses defined the strongest correlation as between deficits in episodic-like memory tasks and total Abeta loads in the brain. Collectively, we find that, in the APPswe/PS1dE9 mouse model, some form of Abeta associated with amyloid deposition can disrupt cognitive circuits when the cholinergic and somatostatinergic systems remain relatively intact; and that episodic-like memory seems to be more sensitive to the toxic effects of Abeta. PMID- 15755687 TI - Role of mitochondria in neuronal cell death induced by oxidative stress; neuroprotection by Coenzyme Q10. AB - Neuronal cells depend on mitochondrial oxidative phosphorylation for most of their energy needs and therefore are at a particular risk for oxidative stress. Mitochondria play an important role in energy production and oxidative stress induced apoptosis. In the present study, we have demonstrated that external oxidative stress induces mitochondrial dysfunction leading to increased ROS generation and ultimately apoptotic cell death in neuronal cells. Furthermore, we have investigated the role of Coenzyme Q10 as a neuroprotective agent. Coenzyme Q10 is a component of the mitochondrial respiratory chain and a potent anti oxidant. Our results indicate that total cellular ROS generation was inhibited by Coenzyme Q10. Further, pre-treatment with Coenzyme Q10 maintained mitochondrial membrane potential during oxidative stress and reduced the amount of mitochondrial ROS generation. Our study suggests that water-soluble Coenzyme Q10 acts by stabilizing the mitochondrial membrane when neuronal cells are subjected to oxidative stress. Therefore, Coenzyme Q10 has the potential to be used as a therapeutic intervention for neurodegenerative diseases. PMID- 15755688 TI - Insulin protects against amyloid beta-peptide toxicity in brain mitochondria of diabetic rats. AB - This study compared the status of brain mitochondria isolated from 12-week streptozotocin (STZ)-diabetic rats versus STZ-diabetic animals treated with insulin during a period of 4 weeks. Brain mitochondria isolated from 12-week citrate (vehicle)-treated rats were used as control. For that purpose, several mitochondrial parameters were evaluated: respiratory indexes (respiratory control ratio (RCR) and ADP/O ratio), transmembrane potential (DeltaPsim), repolarization lag phase, repolarization level, ATP, glutathione and coenzyme Q (CoQ) contents, production of H2O2, ATPase activity, and the capacity of mitochondria to accumulate Ca2+. Furthermore, the effect of Abeta1-40 was also analyzed. We observed that STZ-induced diabetes promoted a significant decrease in mitochondrial CoQ9, ATPase activity, and a lower capacity of mitochondria to accumulate Ca2+ when compared with control and insulin-treated diabetic rats. The presence of 4 microM Abeta1-40 induced a significant decrease in RCR in the three groups of rats. However, this peptide induced a significant increase in the repolarization lag phase and a significant decrease in the repolarization level in control and diabetic animals without insulin treatment. Furthermore, this peptide exacerbated significantly the production of H2O2 in STZ-diabetic rats, this effect being avoided by insulin treatment. Our data show that although diabetes induces some alterations in brain mitochondrial activity, those alterations do not interfere significantly with mitochondria functional efficiency. Similarly, insulin does not affect basal mitochondria function. However, in the presence of amyloid beta-peptide, insulin seems to prevent the decline in mitochondrial oxidative phosphorylation efficiency and avoids an increase in oxidative stress, improving or preserving the function of neurons under adverse conditions, such as Alzheimer's disease. PMID- 15755689 TI - Reduction of Ca2+ stores and capacitative Ca2+ entry is associated with the familial Alzheimer's disease presenilin-2 T122R mutation and anticipates the onset of dementia. AB - Mutations in the presenilin genes PS1 and PS2, the major cause of familial Alzheimer's disease (FAD), are associated with alterations in Ca2+ signalling. In contrast to the majority of FAD-linked PS1 mutations, which cause an overload of intracellular Ca2+ pools, the FAD-linked PS2 mutation M239I reduces Ca2+ release from intracellular stores [Zatti, G., Ghidoni, R., Barbiero, L., Binetti, G., Pozzan, T., Fasolato, C., Pizzo, P., 2004. The presenilin 2 M239I mutation associated with Familial Alzheimer's Disease reduces Ca2+ release from intracellular stores. Neurobiol. Dis. 15/2, 269-278]. We here show that in human FAD fibroblasts another PS2 mutation (T122R) reduces both Ca2+ release and capacitative Ca2+ entry. The observation, done in two monozygotic twins, is of note since only one of the subjects showed overt signs of disease at the time of biopsy whereas the other one developed the disease 3 years later. This finding indicates that Ca2+ dysregulation anticipates the onset of dementia. A similar Ca2+ alteration occurred in HeLa and HEK293 cells transiently expressing PS2 T122R. Based on these data, the "Ca2+ overload" hypothesis in AD pathogenesis is here discussed and reformulated. PMID- 15755690 TI - Platform influence on DNA microarray data in postmortem brain research. AB - In addition to the substantial biological diversity among humans, our limited ability to reliably measure expression changes of small magnitude significantly reduces our capacity to obtain convergent sets of transcriptome data in postmortem brain. In particular, differences in the structure and sensitivity/reproducibility of microarray platforms, and in the variety of tools used to analyze microarray data, strongly influence experimental outcome. In order to better understand the sensitivity, dynamic range, and reproducibility of three common DNA microarray platforms, we compared two human postmortem samples on cDNA microarrays with dual-fluorescence, oligonucleotide GeneChips (Affymetrix), and single-color gel matrix deposited CodeLink oligonucleotide arrays. All three microarray platforms reported a good dynamic range and high correlation in replicate experiments, but they failed to consistently identify the same genes as differentially expressed between the same samples. Given their reproducibility and proven accuracy, different microarray platforms appear to be measuring different things by nature of their design and function. This needs to be taken into account when comparing data across studies. PMID- 15755691 TI - Distinct disease mechanisms in peripheral neuropathies due to altered peripheral myelin protein 22 gene dosage or a Pmp22 point mutation. AB - Point mutations affecting PMP22 can cause hereditary demyelinating and dysmyelinating peripheral neuropathies. In addition, duplication and deletion of PMP22 are associated with Charcot-Marie-Tooth disease Type 1A (CMT1A) and Hereditary Neuropathy with Liability to Pressure Palsy (HNPP), respectively. This study was designed to elucidate disease processes caused by misexpression of Pmp22 and, at the same time, to gain further information on the controversial molecular function of PMP22. To this end, we took advantage of the unique resource of a set of various Pmp22 mutant mice to carry out comparative expression profiling of mutant and wild-type sciatic nerves. Tissues derived from Pmp22-/- ("knockout"), Pmp22tg (increased Pmp22 copy number), and Trembler (Tr; point mutation in Pmp22) mutant mice were analyzed at two developmental stages: (i) at postnatal day (P)4, when normal myelination has just started and primary causative defects of the mutations are expected to be apparent, and (ii) at P60, with the goal of obtaining information on secondary disease effects. Interestingly, the three Pmp22 mutants exhibited distinct profiles of gene expression, suggesting different disease mechanisms. Increased expression of genes involved in cell cycle regulation and DNA replication is characteristic and specific for the early stage in Pmp22-/- mice, supporting a primary function of PMP22 in the regulation of Schwann cell proliferation. In the Tr mutant, a distinguishing feature is the high expression of stress response genes. Both Tr and Pmp22tg mice show strongly reduced expression of genes important for cholesterol synthesis at P4, a characteristic that is common to all three mutants at P60. Finally, we have identified a number of candidate genes that may play important roles in the disease process or in myelination per se. PMID- 15755692 TI - [With gratitude to our referees, three Murats and internet visitors]. PMID- 15755693 TI - [Is slow coronary flow related with inflammation and procoagulant state?]. AB - OBJECTIVE: To investigate the pathogenesis of coronary slow flow (CSF), C reactive protein (CRP) levels as indicator of inflammation and procoagulant activity were studied in patients with CSF. METHODS: Fifty-one patients (22 female, mean age; 53+/-10 years) who were admitted to our clinic with chest pain and had the diagnosis of CSF established by TIMI frame count method and coronary angiography, and 44 healthy subjects (18 female, mean age; 546 years) with normal coronary flow (NCF) were included in the study. Subjects with any infectious and systemic immune disease were excluded from the study. The CRP levels were measured from venous blood samples during admission, at 24th hour and after 3 months in all subjects. Additionally; fibrinogen, plasminogen, plasminogen activator inhibitor (PAI-1), tissue plasminogen activator (t-PA) and von Willebrand factor (vWF) levels were measured to determine the procoagulant activity. RESULTS: There was no significant difference between CRP levels of patients with CSF and healthy subjects during admission (7.26+/-4.2 ng/dl vs. 6.43+/-2.8 ng/dl, p>0.05), at 24th hour (7.84+/-1.3 ng/dl vs. 6.32+/-2.5 ng/dl, p>0.05) and after 3 months (6.37+/-2.4 ng/dl vs. 6.18+/-3.3 ng/dl, p>0.05). There were no differences between levels of CRP when compared according to the TIMI frame count, number of vessels with CSF and artery in which CSF was dominant. Additionally; procoagulant activity assessed by fibrinogen, plasminogen, PAI-1, t PA and vWF levels was similar in both groups. CONCLUSION: Our findings on normal levels of CRP and procoagulant activity, and lack of relation with TIMI frame count made us to think that inflammatory and procoagulant activity did not play a role in the pathogenesis of CSF. PMID- 15755694 TI - The serial changes in plasma homocysteine levels and it's relationship with acute phase reactants in early postmyocardial infarction period. AB - OBJECTIVE: We aimed to study the change in the plasma homocysteine concentration in the early stage of acute myocardial infarction and its relationship with the acute phase reactants. METHODS: We included into the study 33 patients who were admitted to the hospital with acute myocardial infarction within the first three hours after the onset of symptoms. The plasma samples were obtained on admission (within 3 hours onset of symptom) and at 6, 12, 24 hours and 2, 4, 7, 30 and 90th day after admission. RESULTS: The serial homocysteine measurements were as following: 11.87+/-0.71 micromol/L, 11.89+/-0.62 micromol/L, 11.37+/-0.83 micromol/L, 10.96+/-0.93 micromol/L, 11.37+/-0.89 micromol/L, 11.24+/-0.66 micromol/L, 13.09+/-0.64 micromol/L, 12.85+/-0.71 micromol/L, and 12.19+/-0.91 micromol/L, respectively (p=0.05). Statistically significant difference was found only between the hour 24 and the day 7 (p=0.04). However, there was no statistically significant difference between the admission level and none of the other time points. No correlation was identified between acute phase reactants and lipid parameters that were measured serially at the same time periods and homocysteine levels. CONCLUSION: Although homocysteine plasma values obtained during the sixth and twelfth hours of acute myocardial infarction provide reliable results as a risk markers, timing of blood sampling during the myocardial infarction does not have significant role since plasma values of homocysteine did not affect acute phase reactants. PMID- 15755695 TI - Analytical performance of a direct assay for LDL-cholesterol: a comparative assessment versus Friedewald's formula. AB - OBJECTIVE: Because low density lipoprotein-cholesterol (LDL-C) is a modifiable risk factor for coronary artery disease (CAD), its routine measurement is recommended in the evaluation and management of hypercholesterolemia. Concentrations of LDL-C are commonly monitored by means of the Friedewald formula (FF), which provides a relative estimation of LDL-C concentration when the triglyceride (TGs) concentration is <200 mg/dl and there are no abnormal lipids. Because of the limitations of the Friedewald calculation, direct methods for an accurate quantification of LDL-C are needed. METHODS: We critically examined an immunoseparation method for direct assay of LDL-C in a comparison with FF. 1) We measured intraassay and interassay precision using quality-control sera and patient serum pools. Accuracy was evaluated from total error analyses. Sample stability was examined over 2 months. 2) The LDL-C levels obtained with direct assay were compared with those calculated by the FF in 47 randomly chosen patient samples. The samples were classified as group 1 (patients with TGs 60-308 mg/dl, n=25) and group 2 (patients with TGs 320-695 mg/dl, n=22). RESULTS: The direct immunoseparation assay displayed an excellent precision (total coefficient of variance (CV)<2.5%, intraassay CV<1.5% and interassay CV<1.5%). Mean total error was 4.34%. The direct assay met the current National Cholesterol Education Program (NCEP) requirements for LDL-C testing for precision and accuracy. The results of direct method (x) and the FF (y) were highly correlated (r=0.9908, y=1.030 x -0.289, n=25) in group 1, but the results of two methods disagreed (r=0.716, y=0.956 x -24.869, n=22) in group 2 (patients with TGs 320-695 mg/dl). CONCLUSION: The direct immunoseparation assay meets the currently established analytical performance goals and may be useful for the diagnosis and management of hyperlipidemic patients. PMID- 15755696 TI - Effect of octreotide in the prevention of doxorubicin cardiotoxicity. AB - OBJECTIVE: A precise method for prevention from doxorubicin cardiotoxicity is not known. We examined whether octreotide has a protective effect against doxorubicin cardiotoxicity. METHODS: New Zealand rabbits (n=44) were divided into 4 groups according to drugs given: Group A (n=12) doxorubicin and octreotide, Group B (n=12) only doxorubicin, Group C (n=10) only octreotide and Group D (n=10) only saline. Effects of the drugs were evaluated in terms of histopathological score, fractional shortening (FS) and prolongation of the QTc interval. RESULTS: Mean pathological score for cardiotoxicity (Group A: 3.7+/-0.5, Group B: 3.9+/-0.3), prolongation of QTc (Group A: from 244.5+/-21.2 ms to 282.9+/-25.9 ms, p<0.0001; Group B: from 248.5+/-17.7 ms to 298.3+/-13.7 ms, p<0.00001) and the rate of decrease in FS (Group A: from 34.4+/-2.0 to 28.0+/-2.0, p<0.05; Group B: from 35.1+/-1.9 to 24.8+/-1.3, p<0.05) were higher in Group B when compared to Group A, but only difference in the rate of decrease in FS was statistically significant (p<0.001). None of these variables changed significantly in groups C and D. CONCLUSION: In this preliminary study, octreotide seems not to reduce doxorubicin cardiotoxicity. On the other hand, a consistent tendency of decreased cardiotoxicity in octreotide+doxorubicin group was observed, although only the difference in FS decrease was significant. Further investigations are needed to address the issue of the extent and the mechanisms of this effect. PMID- 15755697 TI - [Comparison of multiple prediction models for hypertension (Neural networks, logistic regression and flexible discriminant analyses)]. AB - OBJECTIVE: In this study, we compared performances of logistic regression analysis (LR), flexible discriminant analysis (EAA) and neural networks (SA) in prediction of primary hypertension. METHODS: Predictor variables were family history, lipoprotein A, triglyceride, smoking and body mass index. The data were collected from Cardiology Clinic of Trakya University Medical Faculty in Turkey, 2001. Logistic regression analysis, flexible discriminant analysis and neural networks were used for prediction of control and hypertension groups. Comparison of the performance of all models was done using receiver operating characteristic (ROC) curve analysis. RESULTS: All models had areas under the ROC curve in the range of 0.793-0.984 and SA had sensitivity, specificity, and accuracy greater than 90% at ideal threshold. ROC curve areas of SA and LR, and SA and EAA were statistically different (p<0.001 and p<0.001 respectively), while ROC curve areas of EAA and LR did not differ (p>0.05). CONCLUSION: We concluded that family history, lipoprotein A, triglyceride, smoking and body mass index variables can be used for prediction of control and hypertension groups with statistically better performance of SA over LR and EAA. PMID- 15755698 TI - [Predictive models and hypertension]. PMID- 15755699 TI - [Hemodynamic performance of mechanical aortic valves in narrow aortic annulus cases]. AB - OBJECTIVE: Small-sized prosthetic valves used in aortic valve position usually cause a high transvalvular gradient. This situation, especially in people with a high body surface area, causes a patient-prosthetic valve mismatch. In this study, hemodynamic performances of the two new generation bileaflet, small size mechanical valves were evaluated during rest and stress conditions. METHODS: Eighteen patients with implanted 21-mm St. Jude HP (11 cases) and Sorin Bicarbon (7 cases) prosthetic valves in aortic position were evaluated at rest and during dobutamine infusion, 16.3+/-5.5 months after the operation. Dobutamine infusion was started at a dose of 5 microg/kg/min. The infusion rate was increased every 15 minutes up to 10-20-30 microg/kg/min. Maximum gradient, mean gradient, effective valve area (EVA), effective valve area index (EVAI) and performance index (PI) were calculated during rest and maximum dobutamine infusion rate. RESULTS: When the rest and dobutamine infusion measurements were compared, the maximum and mean gradients were found to be increased moderately, but EVA, EVAI, PI values did not change with increasing heart rates. For St. Jude valve; maximum gradient increased from 30.5+/-6.5 mmHg to 74.7+/-33.6 mmHg (p=0.03) and mean gradient increased from 17.9+/-3.8 to 40.8+/-23.8 mmHg (p=0.03). For Sorin Bicarbon valve; maximum gradient increased from 31.7+/-13.3 mmHg to 52.0+/-11.2 mmHg (p=0.01), mean gradient increased from 16.1+/-6.4 mmHg to 28.8+/-1.0 mmHg (p=0.01). The difference was not significant between the two valves with respect to measured parameters during rest and maximum dobutamine infusion. CONCLUSION: According to our findings, 21-mm Sorin Bicarbon and St. Jude HP mechanical bileaflet valves have good hemodynamic performance during exercise and these types of valves seem to be appropriate for patients with small aortic annulus. PMID- 15755700 TI - [Small aortic annulus and mechanic aortic valves]. PMID- 15755701 TI - [Effects of topical applications of aprotinin and tranexamic acid on blood loss after open heart surgery]. AB - OBJECTIVE: We sought to investigate the effects of topical applications of aprotinin and tranexamic acid in preventing postoperative bleeding during open heart surgery. METHODS: Thirty patients undergoing open-heart surgery with cardiopulmonary bypass were randomized to three different groups. Group 1 (n = 10) received 1000000 KIU aprotinin, Group 2 (n= 10) received 1 gr tranexamic acid and Group 3 (n = 10) received placebo before closure of the sternotomy. During the first three hours and 24 hours total postoperative blood loss and amount of transfused blood products were recorded. RESULTS: In Group 1, postoperative bleeding rates were 122 ml during first 3 hours, 302 ml during 24 hours and 384 ml total. In Group 2, postoperative bleeding rates were 108 ml during first 3 hours, 305 ml during 24 hours and 393 ml total. In control group, bleeding rates were- 162, 347 and 502 ml, respectively. Needs for transfusion were 4.7 U in the aprotinin group, 5.4 U in the tranexamic acid group and 6.0 U in the control group. CONCLUSION: Topical application of aprotinin and tranexamic acid reduced postoperative bleeding, but this was not statistically significant. The differences were insignificant for effectiveness between aprotinin and tranexamic acid and for transfusion requirements. PMID- 15755702 TI - [The effects of obstructive sleep apnea hypopnea syndrome on cardiovascular system]. AB - Obstructive sleep apnea hypopnea syndrome (OSAHS) is characterized by repetitive upper airway obstruction during sleep and it is commonly seen in the adult population, 4% in the men, 2% in the women. The most common nocturnal symptom is snoring while the most common daytime symptom is excessive daytime sleepiness. The gold standard in the diagnosis is polysomnography. Nasal continuous positive airway pressure is the most efficient therapy in the treatment and prevention of the disease. The OSAHS may cause cardiovascular complications in long-term, including systemic hypertension, pulmonary hypertension, congestive heart failure, arrhythmias, stroke and myocardial infarction. All these complications increase the morbidity and mortality of OSAHS. In this paper, effects of OSAHS on cardiovascular system were reviewed. PMID- 15755703 TI - [Cardiovascular manifestations and treatment in Marfan syndrome]. AB - Marfan syndrome is a dominantly inherited connective tissue disease characterized by cardiovascular, skeletal and ocular manifestations, which was firstly described by Antoine Marfan in 1896. The underlying disorder is a mutation, which impairs fibrillin synthesis and is associated with the FBN-1 gene on the 15th chromosome. Ghent Nosology is used for diagnosis. The progressive dilatation of the proximal aorta leading to dissection and rupture is the typical feature of the disease. Aortic aneurysm and aortic rupture are the lethal complications of the disorder. Increased life expectancy depends on the control and the prevention of the cardiovascular complications. The most frequent cardiovascular manifestation is the mitral valve involvement. The principal pathologic findings on the mitral valve are annular dilatation, fibromyxomatous changes of the leaflets and chordae, elongation and rupture of chordae and deposition of calcium. Prolapsus occurs in 80% of the cases. Elective surgery with optimal timing is associated with increased survival. In the follow-up of patients after surgery for aortic aneurysm and dissection, serial imaging studies in order to detect a new onset aneurysm or dissection on different sites of the aorta are essential. PMID- 15755704 TI - [Conditions when coronary angiography should not/is not necessarily be performed]. PMID- 15755705 TI - [Coronary artery fistula from the left circumflex artery to the right bronchial artery]. PMID- 15755706 TI - A combined surgical and endovascular procedure for thoracic aortic aneurysm in a high risk patient. PMID- 15755707 TI - [A case of less invasive closed mitral commissurotomy with aid of transesophageal echocardiography]. PMID- 15755708 TI - Peak pulse pressure during exercise and left ventricular hypertrophy in athletes. PMID- 15755709 TI - [Coronary artery system in a dog]. PMID- 15755710 TI - [The organ effects of systemic inflammation response activated during open heart surgery and current treatment methods]. PMID- 15755711 TI - Hypertrophic cardiomyopathy: the pathological features and the molecular pathogenesis. PMID- 15755712 TI - [Cardiology education]. PMID- 15755713 TI - [Distribution of risk factors according to socioeconomic status in male and female cases with coronary artery disease]. PMID- 15755714 TI - [Evaluation of diastolic function by transmitral color M-mode flow propagation velocity in hypertensive patients]. PMID- 15755715 TI - Surgical treatment of congenital right coronary artery aneurysm and fistula in a patient with previous aortic dissection repair. PMID- 15755716 TI - [Giant left main coronary aneurysm with coronary artery ectasia]. PMID- 15755717 TI - Chest pain, dynamic electrocardiography changes and ventricular arrhythmia in a patient with thoracic disc hernia. PMID- 15755718 TI - Pulmonary valve endocarditis. PMID- 15755720 TI - Isolated left ventricular diverticulum in an adult: dynamic contrast-enhanced MRI findings. PMID- 15755719 TI - Chronic dissective aortic aneurysm as a result of cannulation performed 20 years ago. PMID- 15755721 TI - [A rare case of traumatic dissection and rupture of the distal thoracic aorta]. PMID- 15755722 TI - Protein kinase D specifically mediates apoptosis signal-regulating kinase 1-JNK signaling induced by H2O2 but not tumor necrosis factor. AB - Although both tumor necrosis factor (TNF) and H2O2 induce activation of c-Jun N terminal kinase (JNK) kinase cascades, it is not known whether they utilize distinct intracellular signaling pathways. In this study, we first examined a variety of pharmacological inhibitors on TNF and H2O2-induced JNK activation. Go6983 or staurosporine, which inhibits protein kinase C isoforms had no effects on TNF or H2O2-induced JNK activation. However, Go6976 and calphostin, which can inhibit protein kinase C as well as protein kinase D (PKD), blocked H2O2- but not TNF-induced JNK activation, suggesting that PKD may be specifically involved in H2O2-induced JNK activation. Consistently, H2O2, but not TNF, induced phosphorylation of PKD and translocation of PKD from endothelial cell membrane to cytoplasm where it associates with the JNK upstream activator, apoptosis signal regulating kinase 1 (ASK1). The association is mediated through the pleckstrin homology domain of PKD and the C-terminal domain of ASK1. Inhibition of PKD by Go6976 or by small interfering RNA of PKD blocked H2O2-induced ASK1-JNK activation and endothelial cell apoptosis. Interestingly, H2O2 induced 14-3-3 binding to PKD via the phospho-Ser-205/208 and phospho-Ser-219/223 and H2O2 induced 14-3-3 binding of PKD was specifically blocked by Go6976 but not by Go6983. More significantly, the 14-3-3-binding defective forms of PKD failed to associate with ASK1 and to activate JNK signaling, highlighting the importance of 14-3-3 binding of PKD in H2O2-induced activation of ASK1-JNK cascade. Thus, our data have identified PKD as a critical mediator in H2O2- but not TNF-induced ASK1 JNK signaling. PMID- 15755723 TI - Physical and functional interactions of the lysophosphatidic acid receptors with PDZ domain-containing Rho guanine nucleotide exchange factors (RhoGEFs). AB - Lysophosphatidic acid (LPA) is a serum-derived phospholipid that induces a variety of biological responses in various cells via heterotrimeric G protein coupled receptors (GPCRs) including LPA1, LPA2, and LPA3. LPA-induced cytoskeletal changes are mediated by Rho family small GTPases, such as RhoA, Rac1, and Cdc42. One of these small GTPases, RhoA, may be activated via Galpha(12/13)-linked Rho-specific guanine nucleotide exchange factors (RhoGEFs) under LPA stimulation although the detailed mechanisms are poorly understood. Here, we show that the C terminus of LPA1 and LPA2 but not LPA3 interact with the PDZ domains of PDZ domain-containing RhoGEFs, PDZ-RhoGEF, and LARG, which are comprised of PDZ, RGS, Dbl homology (DH), and pleckstrin homology (PH) domains. In LPA1- and LPA2-transfected HEK293 cells, LPA-induced RhoA activation was observed although the C terminus of LPA1 and LPA2 mutants, which failed to interact with the PDZ domains, did not cause LPA-induced RhoA activation. Furthermore, overexpression of the PDZ domains of PDZ domain-containing RhoGEFs served as dominant negative mutants for LPA-induced RhoA activation. Taken together, these results indicate that formation of the LPA receptor/PDZ domain containing RhoGEF complex plays a pivotal role in LPA-induced RhoA activation. PMID- 15755724 TI - Folding efficiency is rate-limiting in dopamine D4 receptor biogenesis. AB - Dopamine receptors are G protein-coupled receptors that are critically involved in locomotion, reward, and cognitive processes. The D2 class of dopamine receptors (DRD2, -3, and -4) is the target for antipsychotic medication. DRD4 has been implicated in cognition, and genetic studies have found an association between a highly polymorphic repeat sequence in the human DRD4 coding region and attention deficit hyperactivity disorder. Using DRD4 as a model, we show that antipsychotics can function as potent pharmacological chaperones up-regulating receptor expression and can also rescue a non-functional DRD4 folding mutant. This chaperone-mediated up-regulation involves reduced degradation by the 26 S proteasome; likely via the stabilization of newly synthesized receptor in the endoplasmic reticulum. Dopamine itself can function as a chaperone when shuttled into the cell by means of the dopamine transporter. Furthermore, different repeat variants of DRD4 display differential sensitivity to this chaperone effect. These data suggest that folding efficiency may be rate-limiting for dopamine receptor biogenesis and that this efficiency differs between receptor variants. Consequently, the clinical profile of dopaminergic ligands, including antipsychotics, may include their ability to serve as pharmacological chaperones. PMID- 15755725 TI - Interleukin-1beta decreases expression of the epithelial sodium channel alpha subunit in alveolar epithelial cells via a p38 MAPK-dependent signaling pathway. AB - Acute lung injury (ALI) is a devastating syndrome characterized by diffuse alveolar damage, elevated airspace levels of pro-inflammatory cytokines, and flooding of the alveolar spaces with protein-rich edema fluid. Interleukin-1beta (IL-1beta) is one of the most biologically active cytokines in the distal airspaces of patients with ALI. IL-1beta has been shown to increase lung epithelial and endothelial permeability. In this study, we hypothesized that IL 1beta would decrease vectorial ion and water transport across the distal lung epithelium. Therefore, we measured the effects of IL-1beta on transepithelial current, resistance, and sodium transport in primary cultures of alveolar epithelial type II (ATII) cells. IL-1beta significantly reduced the amiloride sensitive fraction of the transepithelial current and sodium transport across rat ATII cell monolayers. Moreover, IL-1beta decreased basal and dexamethasone induced epithelial sodium channel alpha-subunit (alpha ENaC) mRNA levels and total and cell-surface protein expression. The inhibitory effect of IL-1beta on alpha ENaC expression was mediated by the activation of p38 MAPK in both rat and human ATII cells and was independent of the activation of alpha v beta6 integrin and transforming growth factor-beta. These results indicate that IL-1beta may contribute to alveolar edema in ALI by reducing distal lung epithelial sodium absorption. This reduction in ion and water transport across the lung epithelium is in large part due to a decrease in alpha ENaC expression through p38 MAPK dependent inhibition of alpha ENaC promoter activity and to an alteration in ENaC trafficking to the apical membrane of ATII cells. PMID- 15755726 TI - Structure and kinetics of a monomeric glucosamine 6-phosphate deaminase: missing link of the NagB superfamily? AB - Glucosamine 6-phosphate is converted to fructose 6-phosphate and ammonia by the action of the enzyme glucosamine 6-phosphate deaminase, NagB. This reaction is the final step in the specific GlcNAc utilization pathway and thus decides the metabolic fate of GlcNAc. Sequence analyses suggest that the NagB "superfamily" consists of three main clusters: multimeric and allosterically regulated glucosamine-6-phosphate deaminases (exemplified by Escherichia coli NagB), phosphogluconolactonases, and monomeric hexosamine-6-phosphate deaminases. Here we present the three-dimensional structure and kinetics of the first member of this latter group, the glucosamine-6-phosphate deaminase, NagB, from Bacillus subtilis. The structures were determined in ligand-complexed forms at resolutions around 1.4 Angstroms. BsuNagB is monomeric in solution and as a consequence is active (k(cat) 28 s(-1), K(m(app)) 0.13 mM) without the need for allosteric activators. A decrease in activity at high substrate concentrations may reflect substrate inhibition (with K(i) of approximately 4 mM). The structure completes the NagB superfamily structural landscape and thus allows further interrogation of genomic data in terms of the regulation of NagB and the metabolic fate(s) of glucosamine 6-phosphate. PMID- 15755728 TI - Increased protein stability causes DNA methyltransferase 1 dysregulation in breast cancer. AB - We report that DNA methyltransferase 1 (DNMT1) expression is dysregulated in breast cancer. The elevated protein levels are not a result of increased mRNA levels, but rather an increase in protein half-life. We found that DNMT1 protein levels were elevated in breast cancer tissues and in MCF-7 breast cancer cells relative to normal human mammary epithelial cells (HMECs) without a concomitant increase in DNMT1 mRNA or proliferative fraction. Although DNMT1 mRNA levels were properly S-phase-regulated in both cell types, DNMT1 protein levels did not follow S-phase fraction in MCF-7 cells. Rather, an increase in DNMT1 protein stability was found for MCF-7 cells relative to HMECs, and a destruction domain was mapped to the N-terminal 120 amino acids of DNMT1, which was required for its proper ubiquitination and degradation in HMECs. Furthermore, overexpression of DNMT1 with this deleted destruction domain in HMECs resulted in significantly increased genomic 5-methylcytosine levels relative to overexpression of the full length protein. The regulation of DNMT1 destruction via this domain may be dysfunctional in cancer cells leading to subsequent cytosine hypermethylation in the genome. PMID- 15755729 TI - Localization of the N-terminal domain in light-harvesting chlorophyll a/b protein by EPR measurements. AB - The conformational distribution of the N-terminal domain of the major light harvesting chlorophyll a/b protein (LHCIIb) has been characterized by electron electron double resonance yielding distances between spin labels placed in various domains of the protein. Distance distributions involving residue 3 near the N terminus turned out to be bimodal, revealing that this domain, which is involved in regulatory functions such as balancing the energy flow through photosystems (PS) I and II, exists in at least two conformational states. Models of the conformational sub-ensembles were generated on the basis of experimental distance restraints from measurements on LHCIIb monomers and then checked for consistency with the experimental distance distribution between residues 3 in trimers. Only models where residue 3 is located above the core of the protein and extends into the aqueous phase on the stromal side fit the trimer data. In the other state, which consequently is populated only in monomers, the N-terminal domain extends sideways from the protein core. The two conformational states may correspond to two functional states of LHCIIb, namely trimeric LHCIIb associated with PSII in stacked thylakoid membranes and presumably monomeric LHCIIb associated with PSI in nonstacked thylakoids. The switch between these two is known to be triggered by phosphorylation of Thr-6. A similar phosphorylation induced conformational change of the N-terminal domain has been observed by others in bovine annexin IV which, due to the conformational switch, also loses its membrane-aggregating property. PMID- 15755727 TI - Role of photoreceptor-specific retinol dehydrogenase in the retinoid cycle in vivo. AB - The retinoid cycle is a recycling system that replenishes the 11-cis-retinal chromophore of rhodopsin and cone pigments. Photoreceptor-specific retinol dehydrogenase (prRDH) catalyzes reduction of all-trans-retinal to all-trans retinol and is thought to be a key enzyme in the retinoid cycle. We disrupted mouse prRDH (human gene symbol RDH8) gene expression by targeted recombination and generated a homozygous prRDH knock-out (prRDH-/-) mouse. Histological analysis and electron microscopy of retinas from 6- to 8-week-old prRDH-/- mice revealed no structural differences of the photoreceptors or inner retina. For brief light exposure, absence of prRDH did not affect the rate of 11-cis-retinal regeneration or the decay of Meta II, the activated form of rhodopsin. Absence of prRDH, however, caused significant accumulation of all-trans-retinal following exposure to bright lights and delayed recovery of rod function as measured by electroretinograms and single cell recordings. Retention of all-trans-retinal resulted in slight overproduction of A2E, a condensation product of all-trans retinal and phosphatidylethanolamine. We conclude that prRDH is an enzyme that catalyzes reduction of all-trans-retinal in the rod outer segment, most noticeably at higher light intensities and prolonged illumination, but is not an essential enzyme of the retinoid cycle. PMID- 15755730 TI - Na,K-ATPase from mice lacking the gamma subunit (FXYD2) exhibits altered Na+ affinity and decreased thermal stability. AB - The gamma subunit of the Na,K-ATPase, a 7-kDa single-span membrane protein, is a member of the FXYD gene family. Several FXYD proteins have been shown to bind to Na,K-ATPase and modulate its properties, and each FXYD protein appears to alter enzyme kinetics differently. Different results have sometimes been obtained with different experimental systems, however. To test for effects of gamma in a native tissue environment, mice lacking a functional gamma subunit gene (Fxyd2) were generated. These mice were viable and without observable pathology. Prior work in the mouse embryo showed that gamma is expressed at the blastocyst stage. However, there was no delay in blastocele formation, and the expected Mendelian ratios of offspring were obtained even with Fxyd2-/- dams. In adult Fxyd2-/- mouse kidney, splice variants of gamma that have different nephron segment-specific expression patterns were absent. Purified gamma-deficient renal Na,K-ATPase displayed higher apparent affinity for Na+ without significant change in apparent affinity for K+. Affinity for ATP, which was expected to be decreased, was instead slightly increased. The results suggest that regulation of Na+ sensitivity is a major functional role for this protein, whereas regulation of ATP affinity may be context-specific. Most importantly, this implies that gamma and other FXYD proteins have their effects by local and not global conformation change. Na,K ATPase lacking the gamma subunit had increased thermal lability. Combined with other evidence that gamma participates in an early step of thermal denaturation, this indicates that FXYD proteins may play an important structural role in the enzyme complex. PMID- 15755731 TI - Solution X-ray scattering evidence for agonist- and antagonist-induced modulation of cleft closure in a glutamate receptor ligand-binding domain. AB - Agonist-induced conformational changes in the ligand-binding domains (LBD) of glutamate receptor ion channels provide the driving force for molecular rearrangements that mediate channel opening and subsequent desensitization. The resulting regulated transmembrane ion fluxes form the basis for most excitatory neuronal signaling in the brain. Crystallographic analysis of the GluR2 LBD core has revealed a ligand-binding cleft located between two lobes. Channel antagonists stabilize an open cleft, whereas agonists stabilize a closed cleft. The crystal structure of the apo form is similar to the antagonist-bound, open state. To understand the conformational behavior of the LBD in the absence of crystal lattice constraints, and thus better to appreciate the thermodynamic constraints on ligand binding, we have undertaken a solution x-ray scattering study using two different constructs encoding either the core or an extended LBD. In agreement with the GluR2 crystal structures, the LBD is more compact in the presence of agonist than it is in the presence of antagonist. However, the time averaged conformation of the ligand-free core in solution is intermediate between the open, antagonist-bound state and the closed, agonist-bound state, suggesting a conformational equilibrium. Addition of peptide moieties that connect the core domain to the other functional domains in each channel subunit appears to constrain the conformational equilibrium in favor of the open state. PMID- 15755732 TI - BIRB796 inhibits all p38 MAPK isoforms in vitro and in vivo. AB - The compound BIRB796 inhibits the stress-activated protein kinases p38alpha and p38beta and is undergoing clinical trials for the treatment of inflammatory diseases. Here we report that BIRB796 also inhibits the activity and the activation of SAPK3/p38gamma. This occurs at higher concentrations of BIRB796 than those that inhibit p38alpha and p38beta and at lower concentrations than those that inhibit the activation of JNK isoforms. We also show that at these concentrations, BIRB796 blocks the stress-induced phosphorylation of the scaffold protein SAP97, further establishing that this is a physiological substrate of SAPK3/p38gamma. Our results demonstrate that BIRB796, in combination with SB203580, a compound that inhibits p38alpha and p38beta, but not the other p38 isoforms, can be used to identify physiological substrates of SAPK3/p38gamma as well as those of p38alpha and p38beta. PMID- 15755733 TI - Insulin stimulates postsynaptic density-95 protein translation via the phosphoinositide 3-kinase-Akt-mammalian target of rapamycin signaling pathway. AB - Insulin receptors are highly enriched at neuronal synapses, but whose function remains unclear. Here we present evidence that brief incubations of rat hippocampal slices with insulin resulted in an increased protein expression of dendritic scaffolding protein postsynaptic density-95 (PSD-95) in area CA1. This insulin-induced increase in the PSD-95 protein expression was inhibited by the tyrosine kinase inhibitor, AG1024, phosphatidylinositol 3-kinase (PI3K) inhibitors, LY294002 and wortmannin, translational inhibitors, anisomycin and rapamycin, but not by LY303511 (an inactive analogue of LY294002), and transcriptional inhibitor, actinomycin D, suggesting that insulin regulates the translation of PSD-95 by activating the receptor tyrosine kinase-PI3K-mammalian target of rapamycin (mTOR) signaling pathway. A similar insulin-induced increase in the PSD-95 protein expression was detected after stimulation of the synaptic fractions isolated from the hippocampal neurons. Furthermore, insulin treatment did not affect the PSD-95 mRNA levels. In agreement, insulin rapidly induced the phosphorylation of 3-phosphoinositide-dependent protein kinase-1 (PDK1), protein kinase B (Akt), and mTOR, effects that were prevented by the AG1024 and LY294002. We also show that insulin stimulated the phosphorylation of 4E-binding protein 1 (4E-BP1) and p70S6 kinase (p70S6K) in a mTOR-dependent manner. Finally, we demonstrate the constitutive expression of PSD-95 mRNA in the synaptic fractions isolated from hippocampal neurons. Taken together, these findings suggest that activation of the PI3K-Akt-mTOR signaling pathway is essential for the insulin induced up-regulation of local PSD-95 protein synthesis in neuronal dendrites and indicate a new molecular mechanism that may contribute to the modulation of synaptic function by insulin in hippocampal area CA1. PMID- 15755734 TI - Translation of eukaryotic translation initiation factor 4GI (eIF4GI) proceeds from multiple mRNAs containing a novel cap-dependent internal ribosome entry site (IRES) that is active during poliovirus infection. AB - Eukaryotic translation initiation factor 4GI (eIF4GI) is an essential scaffolding protein required to recruit the 43 S complex to the 5'-end of mRNA during translation initiation. We have previously demonstrated that eIF4GI protein expression is translationally regulated. This regulation is mediated by cis acting RNA elements, including an upstream open reading frame and an IRES that directs synthesis of five eIF4GI protein isoforms via alternative AUG initiation codon selection. Here, we further characterize eIF4GI IRES function and show that eIF4GI is expressed from several distinct mRNAs that vary via alternate promoter use and alternate splicing. Several mRNA variants contain the IRES element. We found that IRES activity mapped to multiple regions within the eIF4GI RNA sequence, but not within the 5'-UTR per se. However, the 5'-UTR enhanced IRES activity in vivo and played a role in initiation codon selection. The eIF4GI IRES was active when transfected into cells in an RNA form, and thus, does not require nuclear processing events for its function. However, IRES activity was found to be dependent upon the presence, in cis, of a 5' m7guanosine-cap. Despite this requirement, the eIF4GI IRES was activated by 2A protease cleavage of eIF4GI, in vitro, and retained the ability to promote translation during poliovirus-mediated inhibition of cap-dependent translation. These data indicate that intact eIF4GI protein is not required for the de novo synthesis of eIF4GI, suggesting its expression can continue under stress or infection conditions where eIF4GI is cleaved. PMID- 15755735 TI - Endothelial nitric-oxide synthase antisense (NOS3AS) gene encodes an autophagy related protein (APG9-like2) highly expressed in trophoblast. AB - Macroautophagy is an intracellular degradation system for the majority of proteins and some organelles that is conserved in all eukaryotic species. The precise role of autophagy in mammalian development and potential involvement in disease remain to be discerned. Yeast Atg9p is the first integral membrane protein shown to be essential for the cytoplasm to vacuole targeting (Cvt) pathway and autophagy, whereas its mammalian functional orthologue has yet to be identified. We have identified two human genes homologous to yeast Atg9p and designated these as APG9L1 and APG9L2. We have previously identified APG9L2 as NOS3AS, which participates in the post-transcriptional regulation of the endothelial nitric-oxide synthase (NOS3) gene on chromosome 7 through its antisense overlap. In human adult tissues, APG9L1 was ubiquitously expressed, whereas APG9L2 was highly expressed in placenta (trophoblast cells) and pituitary gland. In transient transfection assays we found that both proteins were primarily localized to the perinuclear region and also scattered throughout the cytosol as dots, a subset of which colocalized with an autophagosome-specific marker LC3 under starvation conditions. Finally, by the small interfering RNA mediated knockdown of APG9L1 in HeLa cells, we demonstrated that APG9L1 is essential for starvation-induced autophagosome formation. In addition, APG9L2 can functionally complement APG9L1 in this process. These results, taken together with those of phylogenetic and sequence analyses, suggest that both APG9L1 and APG9L2 are functionally orthologous to the yATG9 in autophagosome formation. Moreover, APG9L2 is a vertebrate-specific gene that may have gained critical roles in mammalian-specific developmental events, such as placentation, through rapid evolution. PMID- 15755736 TI - Overexpression of the epithelial Na+ channel gamma subunit in collecting duct cells: interactions of Liddle's mutations and steroids on expression and function. AB - The epithelial Na(+) channel (ENaC) has three subunits; the expression of each can be regulated. Liddle's syndrome is caused by an activating mutation in the C terminus of either the beta or gamma subunit. We used a doxycycline-regulated adenovirus system to express varying levels of human gammaENaC in renal collecting duct (M1 cell) monolayers. Increasing levels of wild type human gamma ENaC (gammahENaC) produced a 2.5-fold enhancement of Na(+) transport. Expression of a truncated C terminus produced less protein than wild type or a gammaY627A missense mutation. However, either of these mutations produced a approximately 4 fold increase in Na(+) transport despite the different levels of protein expression. Unexpectedly, overexpression of a marginally detectable amount of gammahENaC was sufficient to produce a full increase in Na(+) transport; a further increase in protein expression produced no further increase in Na(+) transport. Steroid treatment increased Na(+) transport to a similar absolute magnitude in control monolayers and in monolayers expressing all types of gammahENaC. Withdrawal of steroids after 24 h produced a decline in Na(+) transport over 8 h in monolayers expressing wild type but not the Liddle's mutation. Using treatment with brefeldin A to estimate the disappearance rate constants, we found progressively slower disappearance rates in monolayers overexpressing gammahENaC and the Liddle's mutant. Calculated insertion rates were slower for the Liddle's mutant than for wild type despite increasing rates of Na(+) transport. These results raise questions regarding previously held assumptions about the behavior of ENaC. PMID- 15755738 TI - Analysis of ARD1 function in hypoxia response using retroviral RNA interference. AB - Cellular hypoxia response is regulated at the level of hypoxia-inducible factor (HIF) activity. A number of recently identified oxygen sensors are HIF-modifying enzymes that respond to low oxygen by altering HIF modification and thus lead to its activation. In addition to the HIF proline hydroxylases and asparagine hydroxylases, ARD1 is recently described as a HIF-1alpha acetylase that regulates its stability. We found that ARD1 is down-regulated in a number of cell lines in response to hypoxia and hypoxia mimic compounds. After surveying these lines for erythropoietin production and retroviral transfection efficiency, we chose to use HepG2 cells to study the function of ARD1. ARD1 short hairpin RNA delivered by a retroviral vector caused >80% reduction in ARD1 message. We observed decreases in erythropoietin and vascular endothelial growth factor protein production, whereas there was no change in the HIF-1alpha protein level. A gene chip analysis of HepG2 cells transduced with virus expressing ARD1 short hairpin RNA under normoxia and hypoxia conditions or with virus overexpressing recombinant ARD1 confirmed that inhibition of ARD1 does not cause activation of HIF and downstream target genes. However, this analysis revealed that ARD1 is involved in cell proliferation and in regulating a series of cellular metabolic pathways that are regulated during hypoxia response. The role of ARD1 in cell proliferation is confirmed using fluorescence labeling analysis of cell division. From these studies we conclude that ARD1 is not required to suppress HIF but is required to maintain cell proliferation in mammalian cells. PMID- 15755739 TI - Dynamic membrane topology of the Escherichia coli beta-glucoside transporter BglF. AB - The Escherichia coli BglF protein, a permease of the phosphoenolpyruvate dependent phosphotransferase system, catalyzes transport and phosphorylation of beta-glucosides. In addition, BglF regulates bgl operon expression by controlling the activity of the transcriptional regulator BglG via reversible phosphorylation. BglF is composed of three domains; one is hydrophobic, which presumably forms the sugar translocation channel. We studied the topology of this domain by Cys-replacement mutagenesis and chemical modification by thiol reagents. Most Cys substitutions were well tolerated, as demonstrated by the ability of the mutant proteins to catalyze BglF activities. Our results suggest that the membrane domain contains eight transmembrane helices and an alleged cytoplasmic loop that contains two additional helices. The latter region forms a dynamic structure, as evidenced by the alternation of residues near its ends between faced-in and faced-out states. We suggest that this region, together with the two transmembrane helices encompassing it, forms the sugar translocation channel. BglF periplasmic loops are close to the membrane, the first being a reentrant loop. This is the first systematic topological study carried out with an intact phosphotransferase system permease and the first demonstration of a reentrant loop in this group of proteins. PMID- 15755737 TI - Manganese superoxide dismutase protects against 6-hydroxydopamine injury in mouse brains. AB - Dopaminergic neurons of the substantia nigra are susceptible to toxin-based insults. Intrastriatal injection of 6-hydroxydopamine results in selective toxicity to these neurons. A mechanistic role for reactive oxygen species is supported by observations that antioxidants confer protection from 6 hydroxydopamine. Although cell culture studies have suggested extracellular or nonmitochondrial mechanisms in 6-hydroxydopamine toxicity, the compartmentalization of oxidative injury mechanisms is incompletely defined in vivo. Transgenic mice overexpressing mitochondrial manganese superoxide dismutase or extracellular superoxide dismutase received unilateral intrastriatal injections of 6-hydroxydopamine. Mice that overexpress manganese superoxide dismutase showed significantly smaller striatal lesions than littermate controls. There were no differences in nonspecific striatal injury associated with contralateral vehicle injection. Manganese superoxide dismutase overexpression also protected against loss of neuronal cell bodies in the substantia nigra. In contrast, mice overexpressing extracellular superoxide dismutase showed no protection from 6-hydroxydopamine toxicity in either brain region. Protection of the nigrostriatal system by overexpression of manganese superoxide dismutase supports a role for mitochondrially derived superoxide in 6-hydroxydopamine toxicity. Mitochondrial oxidative stress appears to be a common mechanism among diverse models of Parkinson disease, whether involving toxins, mutated genes, or cybrid cells containing patient mitochondria. Antioxidant therapies that target this subcellular compartment may prove promising. PMID- 15755740 TI - Peptide-mediated interference of TIR domain dimerization in MyD88 inhibits interleukin-1-dependent activation of NF-{kappa}B. AB - Myeloid differentiation factor 88 (MyD88) plays a crucial role in the signaling pathways triggered by interleukin (IL)-1 and Toll-like receptors in several steps of innate host defense. A crucial event in this signaling pathway is represented by dimerization of MyD88, which allows the recruitment of downstream kinases like IRAK-1 and IRAK-4. Herein, we have investigated the function of the Toll/IL-1 receptor (TIR) domain in MyD88 homodimerization in cell-free and in vitro experimental settings by using epta-peptides that mimic the BB-loop region of the conserved TIR domain of different proteins. By using a pull-down assay with purified glutathione S-transferase-MyD88 TIR or co-immunoprecipitation experiments, we found that epta-peptides derived from the TIR domain of MyD88 and IL-18R are the most effective in inhibiting homodimerization with either the isolated TIR or full-length MyD88. Moreover, we demonstrated that a cell permeable analog of MyD88 epta-peptide inhibits homodimerization of MyD88 TIR domains in an in vitro cell system and significantly reduces IL-1 signaling, as assayed by activation of the downstream transcription factor NF-kappaB. Our results indicate that the BB-loop in TIR domain of MyD88 is a good target for specific inhibition of MyD88-mediated signaling in vivo. PMID- 15755741 TI - Eap45 in mammalian ESCRT-II binds ubiquitin via a phosphoinositide-interacting GLUE domain. AB - Ubiquitination serves as a key sorting signal in the lysosomal degradation of endocytosed receptors through the ability of ubiquitinated membrane proteins to be recognized and sorted by ubiquitin-binding proteins along the endocytic route. The ESCRT-II complex in yeast contains one such protein, Vps36, which harbors a ubiquitin-binding NZF domain and is required for vacuolar sorting of ubiquitinated membrane proteins. Surprisingly, the presumptive mammalian ortholog Eap45 lacks the ubiquitin-binding module of Vps36, and it is thus not clear whether mammalian ESCRT-II functions to bind ubiquitinated cargo. In this paper, we provide evidence that Eap45 contains a novel ubiquitin-binding domain, GLUE (GRAM-like ubiquitin-binding in Eap45), which binds ubiquitin with similar affinity and specificity as other ubiquitin-binding domains. The GLUE domain shares similarities in its primary and predicted secondary structures to phosphoinositide-binding GRAM and PH domains. Accordingly, we find that Eap45 binds to a subset of 3-phosphoinositides, suggesting that ubiquitin recognition could be coordinated with phosphoinositide binding. Furthermore, we show that Eap45 colocalizes with ubiquitinated proteins on late endosomes. These results are consistent with a role for Eap45 in endosomal sorting of ubiquitinated cargo. PMID- 15755743 TI - Expression of the floral B-function gene SLM2 in female flowers of Silene latifolia infected with the smut fungus Microbotryum violaceum. AB - Silene latifolia is a dioecious plant in which sex is determined by X and Y chromosomes. Expression of the B-function gene SLM2, an ortholog of PISTILLATA (PI) in Arabidopsis, was examined by in situ hybridization. SLM2 was not expressed in suppressed stamens of female flowers, but was expressed in developing stamens of smut-infected female flowers. These results indicate that the control of SLM2 is independent of the presence of the Y chromosome. Smut infected females provide a useful system for clarifying the relationship between the B-function gene and the sex determination factor. PMID- 15755742 TI - Functional characterization of XendoU, the endoribonuclease involved in small nucleolar RNA biosynthesis. AB - XendoU is the endoribonuclease involved in the biosynthesis of a specific subclass of Xenopus laevis intron-encoded small nucleolar RNAs. XendoU has no homology to any known cellular RNase, although it has sequence similarity with proteins tentatively annotated as serine proteases. It has been recently shown that XendoU represents the cellular counterpart of a nidovirus replicative endoribonuclease (NendoU), which plays a critical role in viral replication and transcription. In this paper, we combined prediction and experimental data to define the amino acid residues directly involved in XendoU catalysis. Specifically, we find that XendoU residues Glu-161, Glu-167, His-162, His-178, and Lys-224 are essential for RNA cleavage, which occurs in the presence of manganese ions. Furthermore, we identified the RNA sequence required for XendoU binding and showed that the formation of XendoU-RNA complex is Mn2+-independent. PMID- 15755744 TI - Transcriptional activity of male gamete-specific histone gcH3 promoter in sperm cells of Lilium longiflorum. AB - Histones are essential for packaging of eukaryotic genomic DNA in nucleosomes, and histone gene expression is normally coupled with DNA synthesis. Some of the flowering plant histone genes show strictly male gamete-specific expression. However, mechanisms underlying their male gamete-specific expression have not been elucidated so far. Here we report the isolation of the male gamete-specific histone gcH3 promoter from Lilium longiflorum and its activity in the male gametic cell of the flowering plant. The OCT motif, which is well conserved in plant histone promoters regulating S phase-specific expression, is not conserved in the gcH3 promoter. Instead sequence motifs identical to GC box 1 and GC box 2, the transcriptional activator and suppressor for mammalian testis-specific histone H1t, are present in the gcH3 promoter, suggesting that plants and animals share the mechanism which governs the specificity of gene expression in male gametic cells. Male gamete-specific activation of the gcH3 promoter has been confirmed by microprojectile bombardment in lily pollen. The sperm cell carrying gold particles showed reporter gene expression, while green fluorescent protein (GFP) was absent in the other sperm cell which had no particles, confirming that the gcH3 promoter is activated in the male gametic cell, and sperm cells have transcriptional and translational machinery that is independent of the vegetative cell of pollen. PMID- 15755745 TI - A critical role for the Sp1-binding sites in the transforming growth factor-beta mediated inhibition of lipoprotein lipase gene expression in macrophages. AB - Increasing evidence suggests that the cytokine transforming growth factor-beta (TGF-beta) inhibits the development of atherosclerosis. The lipoprotein lipase (LPL) enzyme expressed by macrophages has been implicated in the pathogenesis of atherosclerosis by stimulating the uptake of lipoprotein particles. Unfortunately, the action of TGF-beta on the expression of LPL in macrophages remains largely unclear. We show that TGF-beta inhibits LPL gene expression at the transcriptional level. Transient transfection assays reveal that the -31/+187 sequence contains the minimal TGF-beta-responsive elements. Electrophoretic mobility shift assays show that Sp1 and Sp3 interact with two regions in the 31/+187 sequence. Mutations of these Sp1/Sp3 sites abolish the TGF-beta-mediated suppression whereas multimers of the sequence impart the response to a heterologous promoter. TGF-beta has no effect on the binding or steady-state polypeptide levels of Sp1 and Sp3. These results, therefore, suggest a novel mechanism for the TGF-beta-mediated repression of LPL gene transcription that involves regulation of the action of Sp1 and Sp3. PMID- 15755746 TI - Regulated chromatin domain comprising cluster of co-expressed genes in Drosophila melanogaster. AB - Recently, the phenomenon of clustering of co-expressed genes on chromosomes was discovered in eukaryotes. To explore the hypothesis that genes within clusters occupy shared chromatin domains, we performed a detailed analysis of transcription pattern and chromatin structure of a cluster of co-expressed genes. We found that five non-homologous genes (Crtp, Yu, CK2betates, Pros28.1B and CG13581) are expressed exclusively in Drosophila melanogaster male germ-line and form a non-interrupted cluster in the 15 kb region of chromosome 2. The cluster is surrounded by genes with broader transcription patterns. Analysis of DNase I sensitivity revealed 'open' chromatin conformation in the cluster and adjacent regions in the male germ-line cells, where all studied genes are transcribed. In contrast, in somatic tissues where the cluster genes are silent, the domain of repressed chromatin encompassed four out of five cluster genes and an adjacent non-cluster gene CG13589 that is also silent in analyzed somatic tissues. The fifth cluster gene (CG13581) appears to be excluded from the chromatin domain occupied by the other four genes. Our results suggest that extensive clustering of co-expressed genes in eukaryotic genomes does in general reflect the domain organization of chromatin, although domain borders may not exactly correspond to the margins of gene clusters. PMID- 15755747 TI - Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts. AB - The expression of angiosperm chloroplast genes is modified by C-to-U RNA editing. The mechanism for recognition of the approximately 30 C targets of editing is not understood. There is no single consensus sequence surrounding editing sites, though sites can be grouped into small 'clusters' of two to five sites exhibiting some sequence similarity. While complementary RNA that guides nucleotides for alteration has been detected in other RNA modification systems, it is not known whether complementary RNA is involved in chloroplast editing site recognition. We investigated the effect of expressing RNA antisense to the sequences -20 to +6 surrounding the RpoB-2 C target of editing, which is a member of a cluster that includes the PsbL-1 and Rps14-1 sites. Previous experiments had shown that chloroplast rpoB transgene transcripts carrying only these 27 nt were edited in vivo at the proper C. Though transcripts carrying sequences -31 to +60 surrounding the RpoB-2 sites were edited in chloroplast transgenic plants, transcripts carrying the -31 to +62 region followed by the 27 nt complementary region were not edited at all. In contrast, a similar construct, in which the C target as well as the preceding and subsequent nucleotides were mismatched within the 27 nt region, was efficiently edited. The presence of any of the four transgenes carrying RpoB-2 sequences in sense and/or antisense orientation resulted in reduced editing at the PsbL-1 site. Chloroplast transgenic plants expressing the three different antisense RNA constructs exhibited abnormal growth and development, though plants expressing the 92 nt sense transcripts were phenotypically normal. PMID- 15755748 TI - Conformational flexibility revealed by the crystal structure of a crenarchaeal RadA. AB - Homologous recombinational repair is an essential mechanism for repair of double strand breaks in DNA. Recombinases of the RecA-fold family play a crucial role in this process, forming filaments that utilize ATP to mediate their interactions with single- and double-stranded DNA. The recombinase molecules present in the archaea (RadA) and eukaryota (Rad51) are more closely related to each other than to their bacterial counterpart (RecA) and, as a result, RadA makes a suitable model for the eukaryotic system. The crystal structure of Sulfolobus solfataricus RadA has been solved to a resolution of 3.2 A in the absence of nucleotide analogues or DNA, revealing a narrow filamentous assembly with three molecules per helical turn. As observed in other RecA-family recombinases, each RadA molecule in the filament is linked to its neighbour via interactions of a short beta-strand with the neighbouring ATPase domain. However, despite apparent flexibility between domains, comparison with other structures indicates conservation of a number of key interactions that introduce rigidity to the system, allowing allosteric control of the filament by interaction with ATP. Additional analysis reveals that the interaction specificity of the five human Rad51 paralogues can be predicted using a simple model based on the RadA structure. PMID- 15755749 TI - Translation of the F protein of hepatitis C virus is initiated at a non-AUG codon in a +1 reading frame relative to the polyprotein. AB - The hepatitis C virus (HCV) genome contains an internal ribosome entry site (IRES) followed by a large open reading frame coding for a polyprotein that is cleaved into 10 proteins. An additional HCV protein, the F protein, was recently suggested to result from a +1 frameshift by a minority of ribosomes that initiated translation at the HCV AUG initiator codon of the polyprotein. In the present study, we reassessed the mechanism accounting for the synthesis of the F protein by measuring the expression in cultured cells of a luciferase reporter gene with an insertion encompassing the IRES plus the beginning of the HCV-coding region preceding the luciferase-coding sequence. The insertion was such that luciferase expression was either in the +1 reading frame relative to the HCV AUG initiator codon, mimicking the expression of the F protein, or in-frame with this AUG, mimicking the expression of the polyprotein. Introduction of a stop codon at various positions in-frame with the AUG initiator codon and substitution of this AUG with UAC inhibited luciferase expression in the 0 reading frame but not in the +1 reading frame, ruling out that the synthesis of the F protein results from a +1 frameshift. Introduction of a stop codon at various positions in the +1 reading frame identified the codon overlapping codon 26 of the polyprotein in the +1 reading frame as the translation start site for the F protein. This codon 26(+1) is either GUG or GCG in the viral variants. Expression of the F protein strongly increased when codon 26(+1) was replaced with AUG, or when its context was mutated into an optimal Kozak context, but was severely decreased in the presence of low concentrations of edeine. These observations are consistent with a Met-tRNA(i)-dependent initiation of translation at a non-AUG codon for the synthesis of the F protein. PMID- 15755750 TI - Characterizing the function and structural organization of the 5' tRNA-like motif within the hepatitis C virus quasispecies. AB - Hepatitis C virus (HCV) RNA is recognized and cleaved in vitro by RNase P enzyme near the AUG start codon. Because RNase P identifies transfer RNA (tRNA) precursors, it has been proposed that HCV RNA adopts structural similarities to tRNA. Here, we present experimental evidence of RNase P sensitivity conservation in natural RNA variant sequences, including a mutant sequence (A368-G) selected in vitro because it presented changes in the RNA structure of the relevant motif. The variation did not abrogate the original RNase P cleavage, but instead, it allowed a second cleavage at least 10 times more efficient, 4 nt downstream from the original one. The minimal RNA fragment that confers sensitivity to human RNase P enzyme was located between positions 299 and 408 (110 nt). Therefore, most of the tRNA-like domain resides within the viral internal ribosome entry site (IRES) element. In the variant, in which the mutation stabilizes a 4 nt stem loop, the second cleavage required a shorter (60 nt) substrate, internal to the minimal fragment substrate, conforming a second tRNA-like structure with similarities to a 'Russian-doll' toy. This new structure did not impair IRES activity, albeit slightly reduced the efficiency of translation both in vitro and in transfected cells. Conservation of the original tRNA-like conformation together with preservation of IRES activity points to an essential role for this motif. This conservation is compatible with the presence of RNA structures with different complexity around the AUG start codon within a single viral population (quasispecies). PMID- 15755752 TI - Heart rate, regularity, and synchronicity in heart failure: a tale of three brothers. PMID- 15755751 TI - Developmentally regulated instability of the GPI-PLC mRNA is dependent on a short lived protein factor. AB - The expression of the vast majority of protein coding genes in trypanosomes is regulated exclusively at the post-transcriptional level. Developmentally regulated mRNAs that vary in levels of expression have provided an insight into one mechanism of regulation; a decrease in abundance is due to a shortened mRNA half-life. The decrease in half-life involves cis-acting elements in the 3' untranslated region of the mRNA. The trans-acting factors necessary for the increased rate of degradation remain uncharacterized. The GPI-PLC gene in Trypanosoma brucei encodes a phospholipase C expressed in mammalian bloodstream form, but not in the insect procyclic form. Here, it is reported that the differential expression of the GPI-PLC mRNA also results from a 10-fold difference in half-life. Second, the instability of the GPI-PLC mRNA in procyclic forms can be reversed by the inhibition of protein synthesis. Third, specifically blocking the translation of the GPI-PLC mRNA in procyclic forms by the inclusion of a hairpin in the 5' untranslated region does not result in stabilization of the mRNA. Thus, the effect of protein synthesis inhibitors in stabilizing the GPI PLC mRNA operates in trans through a short-lived factor dependent on protein synthesis. PMID- 15755753 TI - Pretreatment with corticosteroids attenuates the efficacy of colchicine in preventing recurrent pericarditis: a multi-centre all-case analysis. AB - AIMS: Effective prevention of recurrent pericarditis remains an important yet elusive goal. Corticosteroid therapy often needs to be continued for a prolonged period and causes severe side effects. We performed a multi-centre all-case analysis to investigate the efficacy of colchicine in preventing subsequent relapses of pericarditis, and addressed the hypothesis that pretreatment with corticosteroids may attenuate the beneficial effect of colchicine. METHODS AND RESULTS: One hundred and forty published and unpublished cases of patients treated with colchicine after at least two relapses of pericarditis were aggregated from European centres. Of those, 119 were included in the study group. Only 18% of the patients had relapses under colchicine therapy, and 30% after its discontinuation. There were significantly more relapses among male patients after colchicine treatment (36 vs. 17%, P=0.046), and those with previous corticosteroid treatment (43 vs. 13%, P=0.02). Multivariate logistic regression analysis identified previous corticosteroid therapy (OR 6.68, 95% CI: 1.65-27.02) and male gender (OR 4.20, 95% CI: 1.16-15.21) as independent risk factors for recurrence following colchicine therapy. CONCLUSION: Treatment with colchicine is highly effective in preventing recurrent pericarditis, while pretreatment with corticosteroids exacerbates and extends the course of recurrent pericarditis. PMID- 15755754 TI - Recurrent pericarditis: mysterious or not so mysterious? PMID- 15755755 TI - Model of outcomes of screening mammography: information to support informed choices. AB - OBJECTIVE: To provide easy to use estimates of the benefits and harms of biennial screening mammography for women aged 40, 50, 60, and 70 years. DESIGN: Markov process model, with data from BreastScreen Australia, the Australian Institute of Health and Welfare, and the Australian Bureau of Statistics. MAIN OUTCOME MEASURE: Age specific outcomes expressed per 1000 women over 10 years. RESULTS: For every 1000 women screened over 10 years, 167-251 (depending on age) receive an abnormal result; 56-64 of these women undergo at least one biopsy, 9-26 have an invasive cancer detected by screening, and 3-6 have ductal carcinoma in situ (DCIS) detected by screening. More breast cancers (both invasive and DCIS) are diagnosed among screened than unscreened women. For example, among 1000 women aged 50 who have five biennial screens, 33 breast cancers are diagnosed: 28 invasive cancers (18 detected at screening and 10 interval cancers) and five DCIS (all detected at screening). By comparison, among 1000 women aged 50 who decline screening, 20 cancers are diagnosed over 10 years. There are about 0.5, 2, 3, and 2 fewer deaths from breast cancer over 10 years per 1000 women aged 40, 50, 60, and 70, respectively, who choose to be screened compared with women who decline screening at times determined by relevant policy. CONCLUSION: Benefits and harms of screening mammography are relatively finely balanced. Quantitative estimates such as these can be used to support individual informed choices about screening. PMID- 15755756 TI - Diphenyleneiodium (DPI) reduces oxalate ion- and calcium oxalate monohydrate and brushite crystal-induced upregulation of MCP-1 in NRK 52E cells. AB - BACKGROUND: Our earlier studies have demonstrated upregulation of monocyte chemoattractant protein-1 (MCP-1) in NRK52E rat renal epithelial cells by exposure to oxalate (Ox) ions and crystals of calcium oxalate monohydrate (COM) or the brushite (Br) form of calcium phosphate. The upregulation was mediated by reactive oxygen species (ROS). This study was performed to investigate whether NADPH oxidase is involved in ROS production. METHODS: Confluent cultures of NRK52E cells were exposed to Ox ions or COM and Br crystals. They were exposed for 1, 3, 6, 12, 24 and 48 h for isolation of MCP-1 mRNA and 24 h for enzyme linked immunosorbent assay (ELISA) to determine the secretion of protein into the culture medium. We also investigated the effect of free radical scavenger, catalase, and the NADPH oxidase inhibitor diphenyleneiodium (DPI) chloride, on the Ox- and crystal-induced expression of MCP-1 mRNA and protein. The transcription of MCP-1 mRNA in the cells was determined using real-time polymerase chain reaction. Hydrogen peroxide and 8-isoprostane were measured to investigate the involvement of ROS. RESULTS: Exposure of NRK52E cells to Ox ions as well as the crystals resulted in increased expression of MCP-1 mRNA and production of the chemoattractant. Treatment with catalase reduced the Ox- and crystal-induced expression of both MCP-1 mRNA and protein. DPI reduced the crystal-induced gene expression and protein production but not Ox-induced gene expression and protein production. CONCLUSIONS: Exposure to Ox ions, and COM and Br crystals stimulates a ROS-mediated increase in MCP-1 mRNA expression and protein production. Reduction in ROS production, lipid peroxidation, low-density lipoprotein release, and inducible MCP-1 gene and protein in the presence of DPI indicates an involvement of NADPH oxidase in the production of ROS. PMID- 15755757 TI - Growth hormone induces anabolism in malnourished maintenance haemodialysis patients. AB - BACKGROUND: Growth hormone (GH) promotes anabolism in patients undergoing maintenance haemodialysis (MHD). However, no studies have examined the effects of GH on protein anabolism in MHD patients using full nitrogen-balance techniques. This study tested the hypothesis that recombinant human GH (rhGH) will induce an anabolic response, as assessed by long-term classic nitrogen-balance techniques, in malnourished MHD patients. METHODS: Six adult MHD patients with protein-energy malnutrition underwent nitrogen-balance studies in a general clinical research centre for 28-35 days each. Patients were maintained on a constant dialysis regimen and protein and energy intakes that were similar to their dialysis regimen and diet prior to hospitalization. The first 14-21 hospital days constituted a baseline phase; during the subsequent 8-21 days, patients were given daily subcutaneous injections of rhGH (0.05 mg/kg body weight/day). RESULTS: During treatment with rhGH, serum insulin-like growth factor-I (IGF-I) increased by approximately 225% (P = 0.002), nitrogen balance became strongly positive (+2.35 g/day; P = 0.034 vs baseline) and there was a reduction in serum urea nitrogen (-32%; P = 0.001). Two patients who became acutely ill and had the lowest dietary protein intakes developed a much smaller rise in serum IGF-I levels and increase in nitrogen balance when they received the rhGH treatment. In the remaining four responders, the decrease in nitrogen output was sustained throughout the entire period of treatment with rhGH. There was no change in body weight during the baseline or treatment phases of the study. CONCLUSIONS: Injections of rhGH induce a strong and sustained anabolic effect, as indicated by positive nitrogen balance, in MHD patients with protein-energy malnutrition. This response was attenuated in two patients who were acutely ill with low protein intakes, suggesting that they may have developed partial resistance to GH. PMID- 15755758 TI - Measurement of cyclosporin exposure in renal transplant recipients during the early post-operative period: is C2 alone sufficient? PMID- 15755759 TI - The importance of residual renal function for patients on dialysis. PMID- 15755760 TI - Expression and activation of STAT3 in chronic proliferative immune complex glomerulonephritis and the effect of fosinopril. AB - BACKGROUND: Signal transducers and activators of transcription (STATs) are cytoplasmic proteins that are activated in response to stimulation from various cytokines. Among these, STAT3 is an important member that has been implicated in the inflammatory proliferation of cells. We hypothesized that STAT3 may be activated in kidneys of rats having modified chronic immune complex glomerulonephritis, and that angiotensin-converting enzyme (ACE) inhibition with fosinopril may prevent the activation of STAT3 and subsequent upregulation of tissue inhibitor of metalloproteinase-1 (TIMP-1), which are effects that may explain the therapeutic effects of fosinopril on nephritis. METHODS: Fifty-one Wistar rats were randomly divided into three groups that included a control group, a model group and a fosinopril group. Bovine serum albumin (BSA) nephritis was induced by subcutaneous immunization and daily intraperitoneal (i.p.) administration of BSA. To accentuate the nephritis, we performed uni-nephrectomy and gave 100 microg of lipopolysaccharide (LPS) as an i.p. injection. Macrophage infiltration (ED-1) was assessed with immunohistochemistry. The expression and activation of STAT3 and the expression of TIMP-1, one of the STAT3 downstream genes, were observed in renal tissues of rats by means of immunohistochemistry, electrophoretic mobility shift assay (EMSA), western blot and northern blot. The relationships between STAT3 phosphorylation, 24 h urinary protein excretion and TIMP-1 expression were also analysed. RESULTS: Northern blot showed that the mRNA expression of both STAT3 and TIMP-1 was significantly increased in kidneys from the model group, but significantly decreased in the fosinopril group (P<0.05). Western blot analysis revealed similar increases in the expression of STAT3, phospho-STAT3 (p-STAT3) and TIMP-1 in the model group. Analysis of immunohistochemistry showed that STAT3 and p-STAT3 were expressed in very few cells of normal rats, that expression was strong in model rats and that this increased expression was attenuated in the fosinopril group (P<0.05). The expression of p-STAT3 in glomeruli was positively correlated with 24 h proteinuria as well as with glomerular TIMP-1 expression. Double staining showed that some ED-1-positive cells also contained p-STAT3-positive staining. CONCLUSIONS: The present study showed that STAT3 is expressed and activated in kidneys of rats with modified immune complex glomerulonephritis. These rats also had increased ED-1-positive cells, with some cells showing simultaneous expression of p-STAT3 and ED-1, which may contribute to glomerular inflammatory proliferation and extracellular matrix accumulation. Finally, fosinopril downregulated STAT3 activation and ED-1 influx, which are effects that may attenuate renal damage in this model. PMID- 15755762 TI - Anisocytes and poikilocytes in the urine. PMID- 15755761 TI - Cutaneous Mycobacterium abscessus infection after kidney transplantation. PMID- 15755763 TI - The puzzle of aspirin and sex. PMID- 15755764 TI - Low-density lipoprotein cholesterol in patients with stable coronary heart disease--is it time to shift our goals? PMID- 15755765 TI - Intensive lipid lowering with atorvastatin in patients with stable coronary disease. AB - BACKGROUND: Previous trials have demonstrated that lowering low-density lipoprotein (LDL) cholesterol levels below currently recommended levels is beneficial in patients with acute coronary syndromes. We prospectively assessed the efficacy and safety of lowering LDL cholesterol levels below 100 mg per deciliter (2.6 mmol per liter) in patients with stable coronary heart disease (CHD). METHODS: A total of 10,001 patients with clinically evident CHD and LDL cholesterol levels of less than 130 mg per deciliter (3.4 mmol per liter) were randomly assigned to double-blind therapy and received either 10 mg or 80 mg of atorvastatin per day. Patients were followed for a median of 4.9 years. The primary end point was the occurrence of a first major cardiovascular event, defined as death from CHD, nonfatal non-procedure-related myocardial infarction, resuscitation after cardiac arrest, or fatal or nonfatal stroke. RESULTS: The mean LDL cholesterol levels were 77 mg per deciliter (2.0 mmol per liter) during treatment with 80 mg of atorvastatin and 101 mg per deciliter (2.6 mmol per liter) during treatment with 10 mg of atorvastatin. The incidence of persistent elevations in liver aminotransferase levels was 0.2 percent in the group given 10 mg of atorvastatin and 1.2 percent in the group given 80 mg of atorvastatin (P<0.001). A primary event occurred in 434 patients (8.7 percent) receiving 80 mg of atorvastatin, as compared with 548 patients (10.9 percent) receiving 10 mg of atorvastatin, representing an absolute reduction in the rate of major cardiovascular events of 2.2 percent and a 22 percent relative reduction in risk (hazard ratio, 0.78; 95 percent confidence interval, 0.69 to 0.89; P<0.001). There was no difference between the two treatment groups in overall mortality. CONCLUSIONS: Intensive lipid-lowering therapy with 80 mg of atorvastatin per day in patients with stable CHD provides significant clinical benefit beyond that afforded by treatment with 10 mg of atorvastatin per day. This occurred with a greater incidence of elevated aminotransferase levels. PMID- 15755766 TI - Secretory dynamics of ghrelin in adolescent girls with anorexia nervosa and healthy adolescents. AB - Ghrelin is an orexigenic peptide and a growth hormone (GH) secretagogue. Secretory dynamics of ghrelin have not been characterized in adolescents with anorexia nervosa (AN). We hypothesized that, compared with healthy adolescents, girls with AN would have increased ghrelin concentrations measured over 12 h of nocturnal sampling from increased basal and pulsatile secretion, and endogenous ghrelin would independently predict GH and cortisol. We examined ghrelin concentration and secretory dynamics in 22 girls with AN and 18 healthy adolescents 12-18 yr old. Associations between ghrelin, various hormones, and measures of insulin resistance were examined. On Cluster analysis, girls with AN had higher ghrelin concentrations than controls, including total area under the curve (AUC) (P = 0.002), nadir (P = 0.0006), and valley levels (P = 0.002). On deconvolution analysis, secretory burst amplitude (P = 0.03) and burst mass (P = 0.04) were higher in AN, resulting in higher pulsatile (P = 0.05) and total ghrelin secretion (P = 0.03). Fasting ghrelin independently predicted GH burst frequency (r = 0.44, P = 0.005). The nutritional markers body mass index and body fat predicted postglucose and valley ghrelin but not fasting levels. Ghrelin parameters were inversely associated with fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), leptin, and IGF-I. HOMA-IR was the most significant predictor of most ghrelin parameters. Valley ghrelin independently predicted cortisol burst frequency (52% of variability), and ghrelin parameters independently predicted total triiodothyronine and LH levels. Higher ghrelin concentrations in adolescents with AN are a consequence of increased secretory burst mass and amplitude. The most important predictor of ghrelin concentration is insulin resistance, and ghrelin in turn predicts GH and cortisol burst frequency. PMID- 15755767 TI - Portal 5-hydroxytryptophan infusion enhances glucose disposal in conscious dogs. AB - Intraportal serotonin infusion enhances net hepatic glucose uptake (NHGU) during glucose infusion but blunts nonhepatic glucose uptake and can cause gastrointestinal discomfort and diarrhea at high doses. Whether the serotonin precursor 5-hydroxytryptophan (5-HTP) could enhance NHGU without gastrointestinal side effects during glucose infusion was examined in conscious 42-h-fasted dogs, using arteriovenous difference and tracer ([3-3H]glucose) techniques. Experiments consisted of equilibration (-120 to -30 min), basal (-30 to 0 min), and experimental (EXP; 0-270 min) periods. During EXP, somatostatin, fourfold basal intraportal insulin, basal intraportal glucagon, and peripheral glucose (to double the hepatic glucose load) were infused. In one group of dogs (HTP, n = 6), saline was infused intraportally from 0 to 90 min (P1), and 5-HTP was infused intraportally at 10, 20, and 40 microg x kg(-1) x min(-1) from 90 to 150 (P2), 150 to 210 (P3), and 210 to 270 (P4) min, respectively. In the other group (SAL, n = 7), saline was infused intraportally from 0 to 270 min. NHGU in SAL was 14.8 +/- 1.9, 18.5 +/- 2.3, 16.3 +/- 1.4, and 19.7 +/- 1.6 micromol x kg(-1) x min(-1) in P1-P4, whereas NHGU in 5-HTP averaged 16.4 +/- 2.6, 18.5 +/- 1.4, 20.8 +/- 2.0, and 27.6 +/- 2.6 micromol x kg(-1) x min(-1) (P < 0.05 vs. SAL). Nonhepatic glucose uptake (micromol x kg(-1) x min(-1)) in SAL was 30.2 +/- 4.3, 36.8 +/- 5.8, 44.3 +/- 5.8, and 54.6 +/- 11.8 during P1-P4, respectively, whereas in HTP the corresponding values were 26.3 +/- 6.8, 44.9 +/- 10.1, 47.5 +/- 11.7, and 51.4 +/- 13.2 (not significant between groups). Intraportal 5-HTP enhances NHGU without significantly altering nonhepatic glucose uptake or causing gastrointestinal side effects, raising the possibility that a related agent might have a role in reducing postprandial hyperglycemia. PMID- 15755768 TI - Impact of continuous and pulsatile insulin delivery on net hepatic glucose uptake. AB - The pancreas releases insulin in a pulsatile manner; however, studies assessing the liver's response to insulin have used constant infusion rates. Our aims were to determine whether the secretion pattern of insulin [continuous (CON) vs. pulsatile] in the presence of hyperglycemia 1) influences net hepatic glucose uptake (NHGU) and 2) entrains NHGU. Chronically catheterized conscious dogs fasted for 42 h received infusions including peripheral somatostatin, portal insulin (0.25 mU x kg(-1) x min(-1)), peripheral glucagon (0.9 ng x kg(-1) x min( 1)), and peripheral glucose at a rate double the glucose load to the liver. After the basal period, insulin was infused for 210 min at either four times the basal rate (1 mU x kg(-1) x min(-1)) or an identical amount in pulses of 1 and 4 min duration, followed by intervals of 11 and 8 min (CON, 1/11, and 4/8, respectively) in which insulin was not infused. A variable peripheral glucose infusion containing [3H]glucose clamped glucose levels at twice the basal level ( approximately 200 mg/dl) throughout each study. Hepatic metabolism was assessed by combining tracer and arteriovenous difference techniques. Arterial plasma insulin (microU/ml) either increased from basal levels of 6 +/- 1 to a constant level of 22 +/- 4 in CON or oscillated from 5 +/- 1 to 416 +/- 79 and from 6 +/- 1 to 123 +/- 43 in 1/11 and 4/8, respectively. NHGU (-0.8 +/- 0.3, 0.4 +/- 0.2, and -0.9 +/- 0.4 mg x kg(-1) x min(-1)) and net hepatic fractional extraction of glucose (0.04 +/- 0.01, 0.04 +/- 0.01, and 0.05 +/- 0.01 mg x kg(-1) x min(-1)) were similar during the experimental period. Spectral analysis was performed to assess whether a correlation existed between the insulin secretion pattern and NHGU. NHGU was not augmented by pulsatile insulin delivery, and there is no evidence of entrainment in hepatic glucose metabolism. Thus the loss of insulin pulsatility per se likely has little or no impact on the effectiveness of insulin in regulating liver glucose uptake. PMID- 15755769 TI - Interleukin-6 acts as insulin sensitizer on glycogen synthesis in human skeletal muscle cells by phosphorylation of Ser473 of Akt. AB - Previous studies showed an insulin-"desensitizing" action of IL-6 on glycogen synthesis in hepatocytes. We recently found no inhibition of the proximal steps of the insulin signal cascade in human skeletal muscle cells. Because these data indicate a possible tissue-specific effect of IL-6, we investigated the influence of IL-6 on insulin-stimulated glycogen synthesis in these cells. At first, we found that incubation of the cells with 20 ng/ml IL-6 alone induced phosphorylation of Ser473 of Akt, but not of Thr308 time dependently and we observed that IL-6 augments insulin-induced Ser473 and Thr308 phosphorylation in the low nanomolar range of insulin. Moreover, IL-6 increased insulin-stimulated phosphorylation of glycogen synthase kinase-3. Accordingly, IL-6 enhanced glycogen synthesis in the presence of 3 and 10 nM insulin, whereas IL-6 alone had only a marginal effect. IL-6 treatment of C57Bl/6 mice readily stimulated phosphorylation of Ser473 in skeletal muscle. Our result that IL-6 did not induce Ser473 phosphorylation in the liver of these mice suggests a tissue-specific effect. Together, our data demonstrate a novel insulin-sensitizing function of IL 6 on glycogen synthesis in skeletal muscle cells and indicate that IL-6 exerts cell/tissue-specific effects on insulin action. PMID- 15755770 TI - Shielding evaluation of a medical linear accelerator vault in preparation for installing a high-dose rate 252Cf remote afterloader. AB - In support of the effort to begin high-dose rate 252Cf brachytherapy treatments at Tufts-New England Medical Center, the shielding capabilities of a clinical accelerator vault against the neutron and photon emissions from a 1.124 mg 252Cf source were examined. Outside the clinical accelerator vault, the fast neutron dose equivalent rate was below the lower limit of detection of a CR-39 etched track detector and below 0.14 +/- 0.02 muSv h(-1) with a proportional counter, which is consistent, within the uncertainties, with natural background. The photon dose equivalent rate was also measured to be below background levels (0.1 muSv h(-1)) using an ionisation chamber and an optically stimulated luminescence dosemeter. A Monte Carlo simulation of neutron transport through the accelerator vault was performed to validate measured values and determine the thermal-energy to low-energy neutron component. Monte Carlo results showed that the dose equivalent rate from fast neutrons was reduced by a factor of 100,000 after attenuation through the vault wall, and the thermal-energy neutron dose equivalent rate would be an additional factor of 1000 below that of the fast neutrons. Based on these findings, the shielding installed in this facility is sufficient for the use of at least 5.0 mg of 252Cf. PMID- 15755771 TI - Study of radionuclide distribution around Kudankulam nuclear power plant site (Agastheeswaram taluk of Kanyakumari district, India). AB - The activity concentration of primordial radionuclides 238U, 232Th and 40K have been measured in the sand samples of Agastheeswaram taluk of Kanyakumari district using gamma-ray spectrometer. The average activity of 232Th, 238U and 40K are found to be 5787.1, 1082.9 Bq kg(-1) and BDL, respectively. The total average absorbed dose rate owing to the presence of 232Th, 238U and 40K is found to be 3900.4 nGy h(-1). The annual effective dose is 4.7 mSv y(-1) and the results are discussed in this paper. PMID- 15755772 TI - First results on patient dose measurements from conventional diagnostic radiology procedures in Serbia and Montenegro. AB - The objective of this work is to assess patient doses for the most frequent X-ray examinations for the first time in Serbia and Montenegro. A total of 510 procedures for 11 different examination categories in 3 general hospitals were analysed. Mean and median entrance surface air kerma (ESAK) and kerma area product (KAP) values followed by mean effective doses were reported. Using X-ray tube output data, ESAK for each radiographic examination was calculated, as well as the effective dose for each patient. Except for chest PA examination, all estimated doses are less than stated European and International Atomic Energy Agency (IAEA) reference levels for simple radiographic examinations. For complex examinations involving fluoroscopy and radiography total KAP was measured and contributions from fluoroscopy and radiography were assessed. The study of KAP confirms that the dose level for complex fluoroscopy investigations is closely related to the technique and individual patient variation in terms of fluoroscopy time and number of radiography exposures. The obtained values are comparable to those reported in the UK. Survey data are aimed aid development of a national quality control and radiation protection programme for medical exposures. PMID- 15755773 TI - A simple method for measuring thoron spatial distributions. AB - A simple but effective method that allows the measurement of the 220Rn spatial distribution in working or living environments using a solid-state detector is presented in this paper. The method is based on measurements of the alpha particles emitted by 216Po (the first 220Rn progeny) directly deposited on the detector surface at different distances from a 220Rn exhalation source. The validity of the method is shown by comparing the results of an experiment, where the 220Rn activity concentration is measured under conditions of diffusion at constant temperature, with finite-element calculations. PMID- 15755774 TI - Estimation of the dose to the nursing infant due to direct irradiation from activity present in maternal organs and tissues. AB - Radionuclides deposited internally in the mother will give rise to a radiation dose in the infant in two ways. The radionuclides may be transferred through milk and give rise to an internal dose in the infant, or the radionuclides may emit photons that are absorbed by the infant, giving rise to an external dose. In this paper, the external dose to the newborn infant caused by direct irradiation was estimated for monoenergetic photons. Voxel models (also called voxel phantoms) of the mother and infant were made in three geometries. These models, consisting of volume elements, or voxels, were designed so that the infant model was placed in the lap, at the breast and on the shoulder of the mother model. The Visual Monte Carlo (VMC) code was used to transport the photons through the voxel models. Source regions for the emitted photons, such as the whole body, the thyroid, the lung, the liver and the skeleton, were chosen. For the validation of the calculation procedure, VMC results were favourably compared with the results obtained by using other Monte Carlo programs and also with the previously published results for specific absorbed fractions. This paper provides estimates of the external dose per photon to the infant for photon energies between 0.05 and 2.5 MeV. The external dose per photon estimates were made for the three geometries and for the sources listed above. The results show that, for the geometry of the nursing infant model at the breast, the highest dose to the infant per photon comes from radionuclides deposited in the mother's liver. For the nursing infant model at the shoulder, the highest dose to the infant per photon comes from radionuclides deposited in the mother's thyroid, and for the nursing infant model in the lap, the highest dose to the infant per photon comes from radionuclides deposited uniformly in the whole body. The dose per photon results were then used to estimate the dose an infant might receive over the lactation period (6 months) due to the incorporation of 1 Bq of a radionuclide by the mother. This information may be used to provide external dose estimates to the infant in the case of a known or suspected radionuclide incorporation by the mother due to, for example, a nuclear medicine procedure. PMID- 15755775 TI - How useful are home safety behaviours for predicting childhood injury? A cohort study. AB - Little work has examined the utility of home safety behaviours in predicting childhood injury. This study examines the relationship between safety behaviours and child injury using a cohort of 1717 families, with 2357 children aged 0-7 years. Safety behaviours, and sociodemographic and family characteristics were measured using a validated questionnaire, and medically attended injuries were ascertained from medical records. Hospital admission rates were lower amongst children from families with fitted and working smoke alarms [incidence rate ratio (IRR) 0.55, 95% confidence interval (CI) 0.31-0.96], who stored sharp objects safely (IRR 0.44, 95% CI 0.23-0.84) and who had fitted stair gates (IRR 0.57, 95% CI 0.31-1.03). Not having a stair gate and not storing sharp objects safely had high sensitivities and negative predictive values for predicting hospital admission. These findings are unlikely to be explained by reductions in the risk of injuries these items are designed to prevent. Families with a range of safety behaviours may also be 'safer' in other ways. Further exploration of factors that may explain lower injury rates in these families is required. Information on safety behaviours may be useful for targeting and monitoring injury prevention activity. PMID- 15755776 TI - Bridging the gap between doctors and policymakers: the use of scientific knowledge in local school health care policy in The Netherlands. AB - BACKGROUND: The decentralization of school health care policy in The Netherlands was followed by an increase in diversity, which was most often not evidence based. This study aims to clarify the use of scientific knowledge in school health care policy-making processes: multi-actor processes in networks, trying to solve certain problems. METHODS: Case-study design in four Municipal Health Service regions, using documents and half-structured interviews as data sources. RESULTS: Scientific knowledge is used by only 42% of the actors in 58% of decision-making rounds in policy-making processes. 'Recent' regional data on health indicators are used more often than 'established' (inter)national knowledge of theoretical models. Mainly school health professionals use knowledge as a resource to influence the policy process. Other actors (e.g. managers and municipalities) use formal power, money or 'initiative' as their main resources. Powerful actors put forward less scientific knowledge than actors in dependent positions. Individual actors with a combined scientific and political frame of reference put forward knowledge most frequently, especially in complex networks with many actors, more than one powerful actor, more than one arena, more than one dominant resource and more than one dominant frame of reference. CONCLUSION: The use of scientific knowledge in school health care policy-making processes can and must be improved. Liaison officers can bridge the gap between doctors and policymakers, especially in complex policy networks. They combine a scientific and a political frame of reference and act upon scientific knowledge as a resource in their efforts to influence the policy-making process. PMID- 15755777 TI - What happens to British veterans when they leave the armed forces? AB - BACKGROUND: Little is known about the factors associated with leaving the armed forces, or what predicts subsequent employment success for veterans. It is likely that there is a complex interaction of adverse social outcomes and mental health status in this group. METHOD: Analysis of existing data from the King's Military Cohort, a large, randomly selected, longitudinal cohort of service personnel, many of whom have now left the armed forces. The sample consisted of 8195 service personnel who served in the armed forces in 1991; a third deployed to the Gulf (1990-91), a third deployed to Bosnia (1992-97) and the final third an 'Era' control group in the Armed Forces in 1991 but not deployed. RESULTS: The majority of service leavers do well after leaving and are in full-time employment. Those with poor mental health during service were more likely to leave and had a greater chance of becoming unemployed after leaving. Mental health problems appear to remain static for veterans after leaving. Veterans of the Gulf War enjoyed more favourable employment outcomes, provided that they came home well. CONCLUSIONS: Only a minority of veterans fare badly after service, even amongst those with active tours of duty behind them. Veterans with mental health problems during service seem to be at higher risk of social exclusion after leaving and therefore these individuals represent an especially vulnerable group of the veteran population. PMID- 15755778 TI - Disaster and associated changes in physical and mental health in older residents. AB - BACKGROUND: Long-term health consequences of disasters have not been studied extensively, one reason amongst others is that no pre-disaster observation is available. This study focuses on an aeroplane crash on an Amsterdam suburb. The ongoing Longitudinal Aging Study Amsterdam has one pre-disaster and several post disaster observations, making it possible to study changes in health, taking pre disaster health characteristics into account. METHODS: Three exposure groups are distinguished: those living within a radius of 1 km from the disaster (initial n=39), those living between a radius of 1 and 2 km from the disaster (initial n=56), and those living in the rest of the city of Amsterdam (initial n=508). Health measures include general health, health in comparison with age peers, functional limitations, disability and cognitive functioning. These measures are based on self-ratings, interviewer observations, or both. RESULTS: Older persons living closest to the disaster area are likely to experience health decline in the wake of a disaster, over and above the health decline that would occur normally with aging. The disaster-associated health decline is small, and most obvious in the ability to perform actions (such as mobility), but is not observed in either disability in daily functioning, nor in self-perceptions of health. Cognitive functioning even shows a short-term improvement. CONCLUSION: These findings suggest substantial resilience in older adults, despite their common health problems. PMID- 15755779 TI - Adapting and disseminating effective public health interventions in another country: towards a systematic approach. AB - BACKGROUND: Public health interventions that have proven effective in one country, are often adapted and disseminated in other countries. However, the process by which effective interventions are chosen for adaptation and dissemination in another country is often not conducted systematically. METHODS: We propose a more systematic approach and describe the main steps that have to be taken in the assessment of the usefulness of effective interventions in another national context. RESULTS: The following steps are proposed. Step 0: Point out the most relevant areas in public health (this is a collaborative effort by policy makers, scientists and practitioners). Step 1: Identification of potentially effective interventions (through systematic literature searches). Step 2: Assessing the levels of evidence and grades of recommendation for adoption. Step 3: Can the results of the trials be generalized to the national situation? Step 4: Can the intervention be implemented in the national situation? CONCLUSION: A more systematic approach to the adaptation and dissemination can be adopted. The basic steps described should be worked out in more detail before they can be used in practice. PMID- 15755781 TI - Socioeconomic status and smoking: analysing inequalities with multiple indicators. AB - BACKGROUND AND AIMS: Socioeconomic differences in smoking have been well established. While previous studies have mostly relied on one socioeconomic indicator at a time, this study examined socioeconomic differences in smoking by using several indicators that reflect different dimensions of socioeconomic position. DATA AND METHODS: Data derive from Helsinki Health Study baseline surveys conducted among the employees of the City of Helsinki in 2000 and 2001. The data include 6243 respondents aged 40-60 years (response rate 68%). Six socioeconomic indicators were used: education, occupational status, household income per consumption unit, housing tenure, economic difficulties and economic satisfaction. Their associations with current smoking were examined by fitting sequential logistic regression models. RESULTS: All socioeconomic indicators were strongly associated with smoking among both men and women. When the indicators were examined simultaneously their associations with smoking attenuated, especially when education and occupational status were considered together, and when income and housing tenure were introduced into the models already containing education and occupational status. After mutual adjustment for all socioeconomic indicators, housing tenure and economic satisfaction remained associated with smoking in men. In women, all indicators except income and economic difficulties were inversely associated with smoking after adjustments. CONCLUSIONS: Smoking was associated with structural, material as well as perceived dimensions of socioeconomic disadvantage. Attempts to reduce smoking among the socioeconomically disadvantaged need to target several dimensions of socioeconomic position. PMID- 15755782 TI - Bullying and symptoms among school-aged children: international comparative cross sectional study in 28 countries. AB - BACKGROUND: There have been no large-scale international comparisons on bullying and health among adolescents. This study examined the association between bullying and physical and psychological symptoms among adolescents in 28 countries. METHODS: This international cross-sectional survey included 123,227 students 11, 13 and 15 years of age from a nationally representative sample of schools in 28 countries in Europe and North America in 1997-98. The main outcome measures were physical and psychological symptoms. RESULTS: The proportion of students being bullied varied enormously across countries. The lowest prevalence was observed among girls in Sweden (6.3%, 95% CI: 5.2-7.4), the highest among boys in Lithuania (41.4%, 95% CI 39.4-43.5). The risk of high symptom load increased with increasing exposure to bullying in all countries. In pooled analyses, with sex stratified multilevel logistic models adjusted for age, family affluence and country the odds ratios for symptoms among students who were bullied weekly ranged from 1.83 (95% CI 1.70-1.97) to 2.11 (95% CI 1.95-2.29) for physical symptoms (headache, stomach ache, backache, dizziness) and from 1.67 (95% CI 1.55-1.78) to 7.47 (95% CI 6.87-8.13) for psychological symptoms (bad temper, feeling nervous, feeling low, difficulties in getting to sleep, morning tiredness, feeling left out, loneliness, helplessness). CONCLUSION: There was a consistent, strong and graded association between bullying and each of 12 physical and psychological symptoms among adolescents in all 28 countries. PMID- 15755786 TI - Alcohol ingestion and topical tacrolimus: a disulfiram-like interaction? PMID- 15755787 TI - Rimonabant--a selective CB1 antagonist. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and safety of rimonabant, a new selective cannabinoid receptor antagonist. DATA SOURCES: Primary literature and review articles were obtained via a MEDLINE search (1966-November 2004) using the key terms obesity, smoking cessation, cannabinoid, rimonabant, SR 141716, and SR 141716a. Additional studies and abstracts were identified from the bibliographies of reviewed literature. STUDY SELECTION AND DATA EXTRACTION: Studies and review articles related to rimonabant and the endocannabinoid system were reviewed. Data pertinent to this article were included. DATA SYNTHESIS: Rimonabant is a selective cannabinoid receptor antagonist. Recent data have demonstrated beneficial effects of rimonabant in obesity, smoking cessation, and metabolic syndrome. Animal studies using rimonabant have shown a positive role for reducing hunger, caloric intake, and body weight and in increasing satiety. In humans, rimonabant appears to be effective for treatment of obesity and smoking cessation. Ongoing studies will examine the effect of rimonabant on obesity, metabolic syndrome, smoking cessation, and alcohol abuse. To date, the incidence of adverse effects with rimonabant has been slightly greater than placebo, with the most common being nausea. CONCLUSIONS: Rimonabant appears to be a promising drug in an entirely new class called selective cannabinoid CB1 receptor antagonists. The drug may be approved for treatment of obesity and smoking cessation in 2005. Additional studies are ongoing that may provide information on other clinical uses for this medication. PMID- 15755788 TI - Do authors of drug therapy review articles perform a complete search of the literature? PMID- 15755789 TI - Satisfaction with pharmacotherapy for approved and off-label indications--a Delphi study. AB - BACKGROUND: Prescribing for non-approved uses is widespread in the treatment of AIDS, cancer, and pediatric illnesses, but it is by no means limited to these areas. Few studies have been performed evaluating reasons for off-label prescribing. OBJECTIVE: To explore the satisfaction with drug therapy as one of the potential reasons for off-label uses by testing a hypothesis that the satisfaction with drug therapy for off-label indications is lower than for approved indications. METHODS: The study compared the satisfaction with drug therapy for known off-label indications with a control group of approved indications. Twenty-four of the first 50 single-ingredient drugs, according to their share in the drug cost budget of the Slovenian Compulsory Health Insurance, had 86 different off-label indications eligible for inclusion. A control group of 86 approved indications was randomly selected from the list of all possible approved indications for the same 24 drugs. A 2-round Delphi technique, involving an expert panel of physicians who are members of the drug regulatory agency, was used to evaluate the satisfaction with drug therapy for selected indications. RESULTS: After the second round of the Delphi study, the median scores of satisfaction with drug therapy for approved and off-label indications were 7.00 and 6.50, respectively (p = 0.001). CONCLUSIONS: The study shows that the satisfaction with available drug therapy for off-label indications is lower than for approved indications. The statistical association, biologic plausibility, and coherence with existing information, as well as the temporality of the association, provide supporting evidence that low satisfaction with drug therapy is one of the incentives for off-label use. PMID- 15755790 TI - Decreased warfarin effect after initiation of high-protein, low-carbohydrate diets. AB - OBJECTIVE: To report 2 cases of decreased international normalized ratio (INR) after initiation of a high-protein, low-carbohydrate diet. CASE SUMMARIES: Case 1. A 67-year-old white woman had been receiving warfarin for 3 years for venous thromboembolism. After initiation of a high-protein, low-carbohydrate diet, the patient required a 22.2% increase (from 45 to 57.5 mg/wk) in warfarin dose. Her INR remained in the therapeutic range on this dose for 8 weeks. When the patient stopped the high-protein, low-carbohydrate diet, a decrease back to the original warfarin dose was required to return to a therapeutic INR. Case 2. A 58-year-old white man had been receiving warfarin for 8 years for a cerebrovascular accident. Initiation of a high-protein, low-carbohydrate diet resulted in a 30% increase (from 26.25 to 37.5 mg/wk) in warfarin dose. His warfarin dose was reduced to the original dose after he stopped the high-protein, low-carbohydrate diet. DISCUSSION: The Naranjo probability scale indicated a possible adverse effect between warfarin and high-protein diets. High-protein diets have been shown to increase serum albumin levels. This may result in more warfarin binding to serum albumin, thereby decreasing the anticoagulant effect of warfarin. The increase of albumin occurs rapidly after initiation of a high-protein diet and appears to promptly affect anticoagulation therapy with warfarin. CONCLUSIONS: These cases indicate a significant interaction between high-protein, low-carbohydrate diets and warfarin therapy. Patients receiving warfarin therapy should be educated on and monitored for the potential interaction that occurs with warfarin therapy and high-protein, low-carbohydrate diets. PMID- 15755791 TI - Staggered transition to epoprostenol from treprostinil in pulmonary arterial hypertension. AB - OBJECTIVE: To describe a successful transition process from subcutaneous treprostinil to intravenous epoprostenol after the failure of treprostinil in a patient with idiopathic pulmonary arterial hypertension and present an algorithm to achieve the conversion without significant adverse reactions. CASE SUMMARY: A 25-year-old white female receiving subcutaneous treprostinil 97 ng/kg/min was admitted to the intensive care unit for transition from subcutaneous treprostinil to a target intravenous epoprostenol dose of 72 ng/kg/min via a staggered interval dose adjustment approach. The patient experienced facial flushing, hot flashes, and headache when dose adjustments of the drugs were made simultaneously; however, when dose adjustments were staggered, the adverse reactions did not occur and larger adjustments could be achieved. DISCUSSION: This case demonstrates a suboptimal therapeutic response to treprostinil for the treatment of idiopathic pulmonary arterial hypertension. The transition of treprostinil to epoprostenol is rare; however, in the event therapy change is needed, dosing information is minimal. A staggered transition dosing regimen that accounts for the pharmacokinetic differences between epoprostenol and treprostinil was successfully used in this case. CONCLUSIONS: The approach in this case demonstrates the success of staggered-interval dose adjustments to minimize supratherapeutic symptoms and coincides with the pharmacokinetic profile of the 2 medications. PMID- 15755792 TI - Prokinetic drug therapy in children: a review of current options. AB - OBJECTIVE: To review the pharmacology, safety, and efficacy of the prokinetic agents metoclopramide and erythromycin in children. DATA SOURCES: English language literature was accessed using MEDLINE (1970-June 2004) with metoclopramide, erythromycin, macrolides, gastroesophageal reflux, and gastrointestinal motility as the search terms. STUDY SELECTION AND DATA EXTRACTION: Abstracts and original research articles were included. Preference was given to published controlled trials. Articles providing descriptions of pharmacology, safety, and effectiveness of metoclopramide and erythromycin for the treatment of gastroesophageal reflux (GER) were also used in this review. DATA SYNTHESIS: Some authors advocate using a prokinetic agent along with acid suppression for treatment of GER in children. The 2 prokinetic agents most commonly used are erythromycin and metoclopramide. Erythromycin has numerous observational reports and controlled trials demonstrating its efficacy in improving feeding tolerance in children. Adverse drug reactions associated with its use were uncommon in prospective controlled trials. Few data support the use of metoclopramide for management of GER, and the potential adverse effects associated with its use need to be considered before prescribing. CONCLUSIONS: The literature supports the use of erythromycin as a prokinetic agent. Many children with GER are adequately controlled with acid suppression alone; however, if use of a prokinetic agent is warranted, erythromycin in combination with acid suppression should be considered. Given the lack of prospective controlled studies demonstrating metoclopramide's efficacy and safety in the treatment of GER in children, metoclopramide should not be considered a treatment option. PMID- 15755793 TI - Discovery, clinical development, and therapeutic uses of bisphosphonates. AB - OBJECTIVE: To review the literature concerning the history, development, and therapeutic uses of bisphosphonates. DATA SOURCES: English-language articles were identified through a search of MEDLINE (through December 2004) using the key word bisphosphonate. Reference lists of pivotal studies, reviews, and full prescribing information for the approved agents were also examined. STUDY SELECTION AND DATA EXTRACTION: Selected studies included those that discussed the discovery and initial applications of bisphosphonates, as well as their historical development, pharmacokinetic and pharmacodynamic properties, and current therapeutic uses. DATA SYNTHESIS: Bisphosphonates structurally resemble pyrophosphates (naturally occurring polyphosphates) and have demonstrated similar physicochemical effects to pyrophosphates. In addition, bisphosphonates reduce bone turnover and resist hydrolysis when administered orally. The information gained from initial work with etidronate generated a considerable scientific effort to design new and more effective bisphosphonates. The PCP moiety in the general bisphosphonate structure is essential for binding to hydroxyapatite and allows for a number of chemical variations by changing the 2 lateral side chains (designated R(1) and R(2)). The R(1) side chain determines binding affinity to hydroxyapatite, and the R(2) side chain determines antiresorptive potency. Accordingly, each bisphosphonate has its own characteristic profile of activity. CONCLUSIONS: The bisphosphonates reduce bone turnover, increase bone mass, and decrease fracture risk and therefore have a significant place in the management of skeletal disorders including osteoporosis, Paget's disease, bone metastases, osteogenesis imperfecta, and heterotopic ossification. PMID- 15755794 TI - Pemetrexed therapy for malignant pleural mesothelioma. AB - OBJECTIVE: To review pemetrexed, a novel multi-targeted antifolate agent. DATA SOURCES: A literature search was conducted (1985-September 2004) using MEDLINE and CANCERLIT. Recent abstracts from the American Society of Clinical Oncology were also included, along with the manufacturer's information. Key words were pemetrexed, LY-231514, Alimta, multi-targeted antifolate, malignant pleural mesothelioma. STUDY SELECTION AND DATA EXTRACTION: Relevant information on pharmacology, pharmacokinetics, and safety and efficacy of pemetrexed from clinical trials was selected. DATA SYNTHESIS: Pemetrexed inhibits folate metabolism and purine/pyrimidine synthesis. Based on Phase I and II trials, pemetrexed has antitumor activity in solid tumors such as lung, colorectal, and cervical. A pivotal Phase III study in patients with malignant pleural mesothelioma (MPM) demonstrated survival superiority of pemetrexed-cisplatin regimen versus cisplatin. CONCLUSIONS: Pemetrexed is a promising new drug for the treatment of solid malignancies, most notably MPM. PMID- 15755795 TI - Pharmacologic treatment of opioid-induced sedation in chronic pain. AB - OBJECTIVE: To review the literature for pharmacologic management of opioid induced sedation (OIS) in patients with chronic pain. DATA SOURCES: A search of MEDLINE (1966-October 2004) for English-language literature and selected bibliographies was completed. Search terms included pain, opioid, sedation, psychostimulants, amphetamines, modafinil, and donepezil. DATA SYNTHESIS: Amphetamines and amphetamine-like agents, caffeine, donepezil, and modafinil have been evaluated for OIS. Available literature is limited by numbers of subjects, duration, and trial design; however, there is limited support for the use of methylphenidate, donepezil, and modafinil. CONCLUSIONS: Pharmacologic treatment of OIS should be utilized selectively, given the available literature. Methylphenidate, donepezil, and modafinil may be considered in appropriate patients. PMID- 15755796 TI - Factors associated with celecoxib and rofecoxib utilization. AB - BACKGROUND: The cyclooxygenase-2 (COX-2) selective nonsteroidal antiinflammatory drugs (NSAIDs) celecoxib and rofecoxib (before its removal) are marketed as having fewer gastrointestinal (GI)-related complications than nonselective NSAIDs. However, adverse reaction data suggest that the use of COX-2 selective NSAIDs is associated with clinically significant GI events. OBJECTIVE: To assess whether patients receiving celecoxib and rofecoxib have a greater underlying disease burden than patients prescribed nonselective NSAIDs. METHODS: The study population consisted of members of 11 health plans, aged >34 years, with a pharmacy claim for celecoxib or rofecoxib or a nonselective NSAID dispensed between February 1, 1999, and July 31, 2001, who had been continuously enrolled for >364 days before the dispensing date. Celecoxib and rofecoxib patients were randomly selected without replacement from a pool of eligible users in each of the 30 months. Nonselective NSAID users were randomly chosen without replacement within each month on a 2:1 ratio to cases; they could be chosen in more than one month. Univariate analyses comparing 9000 cases and 18 000 controls were performed, followed by a multiple logistic regression analysis conditioned on time. RESULTS: Increasing age, treatment by a rheumatologist or an orthopedic specialist, treatment with a high number of different medications in the past year, treatment with oral corticosteroids in the past year, and having had a previous GI bleed increased the likelihood of receiving celecoxib or rofecoxib, whereas treatment with a high number of nonselective NSAID prescriptions in the past year decreased it. Treatment with a high number of different medications was a predictor of increased prevalence of underlying diabetes mellitus and cardiovascular disease. CONCLUSIONS: Patients having a greater underlying disease burden were more likely to receive COX-2 selective NSAIDs than nonselective ones. Paradoxically, patients at higher risk for cardiovascular disease were channeled toward treatment with COX-2 selective NSAIDs, many of which may confer an increased risk of acute myocardial infarction and other adverse cardiovascular outcomes. PMID- 15755798 TI - Functional INAD complexes are required to mediate degeneration in photoreceptors of the Drosophila rdgA mutant. AB - The TRP family of ion channels mediates a wide range of calcium-influx phenomena in eukaryotic cells. Many members of this family are activated downstream of phosphoinositide hydrolysis but the subsequent steps that lead to TRP channel activation in vivo remain unclear. Recently, the lipid products of phosphoinositide hydrolysis (such as diacylglycerol and its metabolites) have been implicated in activating TRP channels in both Drosophila and mammals. In Drosophila photoreceptors, lack of diacylglycerol kinase (DGK) activity (encoded by rdgA) leads to both constitutive TRP-channel activity and retinal degeneration. In this study, using a novel forward-genetic screen, we identified InaD, a multivalent PDZ domain protein as a suppresser of retinal degeneration in rdgA mutants. We show that InaD suppresses rdgA and that the rescue is correlated with reduced levels of phospholipase Cbeta (PLCbeta), a key enzyme for TRP channel activation. Furthermore, we show that light, Gq and PLCbeta all modulate retinal degeneration in rdgA. The results demonstrate a previously unknown requirement for a balance of PLCbeta and DGK activity for retinal degeneration in rdgA. They also suggest a key role for the lipid products of phosphoinositide hydrolysis in the activation of TRP channels in vivo. PMID- 15755797 TI - Nuclear localization of HTLV-I bZIP factor (HBZ) is mediated by three distinct motifs. AB - The genome of the human T-cell leukemia virus type I (HTLV-I) codes for a basic leucine zipper protein, HBZ, capable of repressing JUN activity and viral transcription. Transient expression in mammalian cells showed that HBZ was targeted to the nucleus, where it accumulated in nuclear speckles. By using a complementary set of deletion mutants, we report here that the nuclear targeting of HBZ is mediated by three distinct nuclear localization signals and that at least two are necessary for the translocation of HBZ to the nucleus. Moreover, the resulting mutant proteins distribute throughout the nucleoplasm and/or into the nucleoli, whereas the wild-type HBZ exclusively accumulates in nuclear speckles, suggesting that the integrity of the protein is required for its speckle localization. We also demonstrate that the HBZ-containing speckles do not correspond to Cajal bodies, splicing factor compartments, or promyelocytic leukemia oncoprotein bodies. Unexpectedly, by using immunogold electron microscopy, we found HBZ localized to heterochromatin. Until now, such characteristics had never been described for a transcription factor and could explain the inhibitory activity of HBZ. PMID- 15755800 TI - Resistance to the antibiotic Zeocin by stable expression of the Sh ble gene does not fully suppress Zeocin-induced DNA cleavage in human cells. AB - Zeocin is a member of the bleomycin/phleomycin family of antibiotics, known to bind and cleave DNA. We established human SK-OV-3 cells that stably express the Zeocin resistance gene (Sh ble) using an ecdysone-inducible mammalian expression system. Surprisingly, our results demonstrated that Zeocin, added in the culture medium to maintain the expression of the ecdysone receptor, was responsible for the formation of DNA strand breaks in the recombinant cells. This suggests that the Zeocin is not completely detoxified and is still able to cleave DNA, despite the stable expression of the Sh ble gene in the recombinant clones. Our study indicates that one needs to be very cautious in the interpretation of data involving stable cell lines selected with Zeocin. PMID- 15755799 TI - O-glycosylation is essential for intracellular targeting of synaptotagmins I and II in non-neuronal specialized secretory cells. AB - We have examined the trafficking of synaptotagmin (Syt) I and II in the mast cell line rat basophilic leukemia (RBL-2H3). We demonstrate that both Syt I and Syt II travel through the plasma membrane and require endocytosis to reach their final intracellular localization. However, N- or C-terminal tagging of Syt II, but not of Syt I, prevents its internalization, trapping the tagged protein at the plasma membrane. Furthermore, a chimeric protein comprising a tagged luminal domain of Syt II fused with the remaining domains of Syt I also localizes to the plasma membrane, whereas a chimera consisting of tagged luminal domain of Syt I fused with Syt II colocalizes with Syt I on secretory granules. We also show that endocytosis of both Syt I and Syt II is strictly dependent on O-glycosylation processing, whereby O-glycosylation mutants of either protein fail to internalize and remain at the plasma membrane. Our results indicate that the luminal domains of Syt I and Syt II govern their internalization capacity from the plasma membrane and identify O-glycosylation as playing a crucial role in Syt trafficking in non-neuronal secretory cells. PMID- 15755801 TI - Hepatic DNA adduct dosimetry in rats fed tamoxifen: a comparison of methods. AB - Liver homogenates from rats fed tamoxifen (TAM) in the diet were shared among four different laboratories. TAM-DNA adducts were assayed by high pressure liquid chromatography-electrospray tandem mass spectrometry (HPLC-ES-MS/MS), TAM-DNA chemiluminescence immunoassay (TAM-DNA CIA), and (32)P-postlabeling with either thin layer ((32)P-P-TLC) or liquid chromatography ((32)P-P-HPLC) separation. In the first study, rats were fed a diet containing 500 p.p.m. TAM for 2 months, and the values for measurements of the (E)-alpha-(deoxyguanosin-N(2)-yl)-tamoxifen (dG-N(2)-TAM) adduct in replicate rat livers varied by 3.5-fold when quantified using 'in house' TAM-DNA standards, or other approaches where appropriate. In the second study, rats were fed 0, 50, 250 or 500 p.p.m. TAM for 2 months, and TAM DNA values were quantified using both 'in house' approaches as well as a newly synthesized [N-methyl-(3)H]TAM-DNA standard that was shared among all the participating groups. In the second study, the total TAM-DNA adduct values varied by 2-fold, while values for the dG-N(2)-TAM varied by 2.5-fold. Ratios of dG-N(2) TAM:(E)-alpha-(deoxyguanosin-N(2)-yl)-N-desmethyltamoxifen (dG-N(2)-N-desmethyl TAM) in the second study were approximately 1:1 over the range of doses examined. The study demonstrated a remarkably good agreement for TAM-DNA adduct measurements among the diverse methods employed. PMID- 15755802 TI - Mutagenesis studies of the major benzo[a]pyrene N2-dG adduct in a 5'-TG versus a 5'-UG sequence: removal of the methyl group causes a modest decrease in the [G >T/G->A] mutational ratio. AB - The potent mutagen/carcinogen benzo[a]pyrene (B[a]P) is metabolically activated to (+)-anti-B[a]PDE, which induces a full spectrum of mutations primarily at the G:C base pairs (e.g. GC-->TA, GC-->AT, etc.). Each of these mutations can be induced by its major adduct [+ta]-B[a]P-N(2)-dG, where DNA sequence context appears to influence both the quantitative and qualitative pattern of mutagenesis. We noted previously that 5'-TG sequences tend to have a higher fraction of G-->T mutations for both [+ta]-B[a]P-N(2)-dG and (+)-anti-B[a]PDE in comparison with 5'-CG, 5'-GG or 5'-AG sequences. To investigate a possible structural element for this trend, the role (if any) of the methyl group on the 5'-T is considered. Using adduct site-specific means, the [G-->T/G-->A] mutational ratio for [+ta]-B[a]P-N(2)-dG is determined to be approximately 1.08 in a 5'-TGT sequence, and approximately 0.60 in a 5'-UGT sequence. (G-->C mutations are minor.) Although this modest approximately 1.8-fold decrease in [G- >T/G-->A] ratio is statistically significant (P = 0.03), it suggests that the methyl group on the 5'-T is not the main reason why a 5'-T tends to enhance G-->T mutations. This study was prompted by an adduct conformational hypothesis, which predicted that the removal of the methyl group in a 5'-TG sequence would lower the fraction of G-->T mutations; however, the approximately 1.8-fold decrease is too small to do additional experiments to assess whether this conformational hypothesis, or other hypotheses, are the true cause of the decrease, which is discussed in this paper. PMID- 15755803 TI - In vitro effect of karathane LC (dinocap) on human lymphocytes. AB - Karathane LC (active ingredient dinocap), a contact fungicide and a non-systemic acaricide was investigated for its ability to induce chromosome aberrations (CAs) and sister chromatid exchanges (SCEs) in cultured human lymphocytes of peripheral blood. In addition to the cytogenetic analysis, the effect of karathane LC on the cell proliferation kinetics (CPK) by the replication index (RI) was studied. The mitotic index (MI) was also determined to detect the cytotoxic effect. Lymphocytes were treated with four different concentrations (5, 10, 15 and 20 microg/ml) of karathane LC for 24 and 48 h. Significant differences between exposed and non-exposed groups found in CAs, SCEs and MI demonstrate the mutagenic, clastogenic and also the cytotoxic effect of karathane LC. PMID- 15755804 TI - Analysis of mouse embryonic patterning and morphogenesis by forward genetics. AB - Many aspects of the genetic control of mammalian embryogenesis cannot be extrapolated from other animals. Taking a forward genetic approach, we have induced recessive mutations by treatment of mice with ethylnitrosourea and have identified 43 mutations that affect early morphogenesis and patterning, including 38 genes that have not been studied previously. The molecular lesions responsible for 14 mutations were identified, including mutations in nine genes that had not been characterized previously. Some mutations affect vertebrate-specific components of conserved signaling pathways; for example, at least five mutations affect previously uncharacterized regulators of the Sonic hedgehog (Shh) pathway. Approximately half of all of the mutations affect the initial establishment of the body plan, and several of these produce phenotypes that have not been described previously. A large fraction of the genes identified affect cell migration, cellular organization, and cell structure. The findings indicate that phenotype-based genetic screens provide a direct and unbiased method to identify essential regulators of mammalian development. PMID- 15755806 TI - Segmental trisomy of chromosome 17: a mouse model of human aneuploidy syndromes. AB - Triplication of whole autosomes or large autosomal segments is detrimental to the development of a mammalian embryo. The trisomy of human chromosome (Chr) 21, known as Down's syndrome, is regularly associated with mental retardation and a variable set of other developmental anomalies. Several mouse models of Down's syndrome, triplicating 33-104 genes of Chr16, were designed in an attempt to analyze the contribution of specific orthologous genes to particular developmental features. However, a recent study challenged the concept of dosage sensitive genes as a primary cause of an abnormal phenotype. To distinguish between the specific effects of dosage-sensitive genes and nonspecific effects of a large number of arbitrary genes, we revisited the mouse Ts43H/Ph segmental trisomy. It encompasses >310 known genes triplicated within the proximal 30 megabases (Mb) of Chr17. We refined the distal border of the trisomic segment to the interval bounded by bacterial artificial chromosomes RP23-277B13 (location 29.0 Mb) and Cbs gene (location 30.2 Mb). The Ts43H mice, viable on a mixed genetic background, exhibited spatial learning deficits analogous to those observed in Ts65Dn mice with unrelated trisomy. Quantitative analysis of the brain expression of 20 genes inside the trisomic interval and 12 genes lying outside on Chr17 revealed 1.2-fold average increase of mRNA steady-state levels of triplicated genes and 0.9-fold average down-regulation of genes beyond the border of trisomy. We propose that systemic comparisons of unrelated segmental trisomies, such as Ts65Dn and Ts43H, will elucidate the pathways leading from the triplicated sequences to the complex developmental traits. PMID- 15755805 TI - Tbx2b is essential for neuronal differentiation along the dorsal/ventral axis of the zebrafish retina. AB - The mechanisms by which retinal neurons are patterned along the dorsal/ventral axis remain largely unknown, yet this patterning is integral for the topographic mapping of visual space. With an interest in elucidating the mechanisms that regulate the development of this retinal axis, we have characterized a T-box family transcription factor, Tbx2b, during zebrafish retinogenesis. Tbx2b is expressed throughout all phases of retinal development with a striking asymmetry of distribution highest dorsally to lowest ventrally. To examine Tbx2b function during retinal development, two morpholino antisense oligonucleotides were created; one blocking the translational start site of Tbx2b and the other interfering with Tbx2b mRNA splicing. Injection of either of these morpholinos resulted in profound defects in the development of the dorsal retina. By using molecular markers for neuronal subtypes, the ventral retina contained all cell types, whereas in the dorsal retina, only retinal ganglion cells expressed markers of differentiation. The cells of the dorsal retina were postmitotic, however, as demonstrated by a lack of BrdUrd incorporation during the normal periods of retinal differentiation. Markers for dorsal and ventral retinal compartments were also expressed normally in Tbx2b morphants. Combined, these observations suggest that the cellular mechanisms regulating neuronal differentiation within the retina are asymmetric about the dorsal/ventral axis and that Tbx2b mediates this process within the dorsal retina. PMID- 15755807 TI - Effects of titanium particle size on osteoblast functions in vitro and in vivo. AB - The formation of titanium (Ti)-wear particles during the lifetime of an implant is believed to be a major component of loosening due to debris-induced changes in bone cell function. Radiographic evidence indicates a loss of fixation at the implant-bone interface, and we believe that the accumulation of Ti particles may act on the bone-remodeling process and impact both long- and short-term implant fixation strengths. To determine the effects of various sizes of the Ti particles on osteoblast function in vivo, we measured the loss of integration strength around Ti-pin implants inserted into a rat tibia in conjunction with Ti particles from one of four size-groups. Implant integration is mediated primarily by osteoblast adhesion/focal contact pattern, viability, proliferation and differentiation, and osteoclast recruitment at the implant site in vivo. This study demonstrates the significant attenuation of osteoblast function concurrent with increased expression of receptor activator of nuclear factor kappaB ligand (RANKL), a dominant signal for osteoclast recruitment, which is regulated differentially, depending on the size of the Ti particle. Zymography studies have also demonstrated increased activities of matrix metalloproteinases (MMP) 2 and 9 in cells exposed to larger Ti particles. In summary, all particles have adverse effects on osteoblast function, resulting in decreased bone formation and integration, but different mechanisms are elicited by particles of different sizes. PMID- 15755808 TI - On the utility of pooling biological samples in microarray experiments. AB - Over 15% of the data sets catalogued in the Gene Expression Omnibus Database involve RNA samples that have been pooled before hybridization. Pooling affects data quality and inference, but the exact effects are not yet known because pooling has not been systematically studied in the context of microarray experiments. Here we report on the results of an experiment designed to evaluate the utility of pooling and the impact on identifying differentially expressed genes. We find that inference for most genes is not adversely affected by pooling, and we recommend that pooling be done when fewer than three arrays are used in each condition. For larger designs, pooling does not significantly improve inferences if few subjects are pooled. The realized benefits in this case do not outweigh the price paid for loss of individual specific information. Pooling is beneficial when many subjects are pooled, provided that independent samples contribute to multiple pools. PMID- 15755809 TI - Cross GTPase-activating protein (CrossGAP)/Vilse links the Roundabout receptor to Rac to regulate midline repulsion. AB - The regulators of the Rho-family GTPases, GTPase-activating proteins (GAPs) and guanine exchange factors (GEFs), play important roles in axon guidance. By means of a functional genomic study of the Rho-family GEFs and GAPs in Drosophila, we have identified a Rho-family GAP, CrossGAP (CrGAP), which is involved in Roundabout (Robo) receptor-mediated repulsive axon guidance. CrGAP physically associates with the Robo receptor. Too much or too little CrGAP activity leads to defects in Robo-mediated repulsion at the midline choice point. The CrGAP gain-of function phenotype mimics the loss-of-function phenotypes of both Robo and Rac. Dosage-sensitive genetic interactions among CrGAP, Robo, and Rac support a model in which CrGAP transduces signals downstream of Robo receptor to regulate Rac dependent cytoskeletal changes. PMID- 15755810 TI - Functional- and abundance-based mechanisms explain diversity loss due to N fertilization. AB - Human activities have increased N availability dramatically in terrestrial and aquatic ecosystems. Extensive research demonstrates that local plant species diversity generally declines in response to nutrient enrichment, yet the mechanisms for this decline remain unclear. Based on an analysis of >900 species responses from 34 N-fertilization experiments across nine terrestrial ecosystems in North America, we show that both trait-neutral and trait-based mechanisms operate simultaneously to influence diversity loss as production increases. Rare species were often lost because of soil fertilization, randomly with respect to traits. The risk of species loss due to fertilization ranged from >60% for the rarest species to 10% for the most abundant species. Perennials, species with N fixing symbionts, and those of native origin also experienced increased risk of local extinction after fertilization, regardless of their initial abundance. Whereas abundance was consistently important across all systems, functional mechanisms were often system-dependent. As N availability continues to increase globally, management that focuses on locally susceptible functional groups and generally susceptible rare species will be essential to maintain biodiversity. PMID- 15755811 TI - Mate choice decisions of stickleback females predictably modified by MHC peptide ligands. AB - Sexual selection has been proposed as one mechanism to explain the maintenance of high allelic diversity in MHC genes that control the extent of resistance against pathogens and parasites in natural populations. MHC-based sexual selection is known to involve olfactory mechanisms in fish, mice, and humans. During mate choice, females of the three-spined stickleback (Gasterosteus aculeatus) use an odor-based selection strategy to achieve an optimal level of MHC diversity in their offspring, equipping them with optimal resistance toward pathogens and parasites. The molecular mechanism of odor-based mate-selection strategies is unknown. Because peptide ligands for MHC class I molecules function as individuality signals in mice, we hypothesized that female sticklebacks might assess the degree of MHC diversity of potential partners by means of the structural diversity of the corresponding peptide ligands in perceived odor signals. We show that structurally diverse MHC ligands interact with natural odors of male sticklebacks to predictably modify MHC-related mate choice. For a mating pair with suboptimal numbers of MHC alleles, peptides increase the attractiveness of male water, whereas for a mating pair with superoptimal numbers, attractiveness is decreased. Our results suggest that female sticklebacks use evolutionarily conserved structural features of MHC peptide ligands to evaluate MHC diversity of their prospective mating partners. PMID- 15755812 TI - Identification of transcribed sequences in Arabidopsis thaliana by using high resolution genome tiling arrays. AB - Using a maskless photolithography method, we produced DNA oligonucleotide microarrays with probe sequences tiled throughout the genome of the plant Arabidopsis thaliana. RNA expression was determined for the complete nuclear, mitochondrial, and chloroplast genomes by tiling 5 million 36-mer probes. These probes were hybridized to labeled mRNA isolated from liquid grown T87 cells, an undifferentiated Arabidopsis cell culture line. Transcripts were detected from at least 60% of the nearly 26,330 annotated genes, which included 151 predicted genes that were not identified previously by a similar genome-wide hybridization study on four different cell lines. In comparison with previously published results with 25-mer tiling arrays produced by chromium masking-based photolithography technique, 36-mer oligonucleotide probes were found to be more useful in identifying intron-exon boundaries. Using two-dimensional HPLC tandem mass spectrometry, a small-scale proteomic analysis was performed with the same cells. A large amount of strongly hybridizing RNA was found in regions "antisense" to known genes. Similarity of antisense activities between the 25-mer and 36-mer data sets suggests that it is a reproducible and inherent property of the experiments. Transcription activities were also detected for many of the intergenic regions and the small RNAs, including tRNA, small nuclear RNA, small nucleolar RNA, and microRNA. Expression of tRNAs correlates with genome-wide amino acid usage. PMID- 15755813 TI - Core 3 synthase is down-regulated in colon carcinoma and profoundly suppresses the metastatic potential of carcinoma cells. AB - The core 3 structure of the O-glycan, GlcNAcbeta1-3GalNAcalpha1-Ser/Thr, an important precursor in the biosynthesis of mucin-type glycoproteins, is synthesized by beta1,3-N-acetylglucosaminyltransferase 6 (beta3Gn-T6; core 3 synthase). We generated an anti-beta3Gn-T6 mAb (G8-144 mAb) and performed immunohistochemical analyses. In normal stomach and colon, beta3Gn-T6 was strongly expressed in the Golgi region of epithelia. In contrast, its expression was markedly down-regulated in gastric and colorectal carcinomas. Tissue specimens from a familial adenomatous polyposis patient showed a clear correlation between the down-regulation of beta3Gn-T6 expression and the degree of dysplasia/neoplasia. In vitro, the level of beta3Gn-T6 transcript was increased according to the differentiation of Caco-2 cells. These results suggested that the expression of beta3Gn-T6 is closely regulated during differentiation and dedifferentiation. beta3Gn-T6 would be a useful marker for distinguishing between benign adenomas and premalignant lesions. HT1080 FP-10 cells stably transfected with the beta3Gn-T6 gene showed a decrease in the core 1 structure, Galbeta1,3GalNAcalpha1-Ser/Thr, probably due to competition between the core 1 synthase and core 3 synthase. The migration activity of the transfectants was markedly lower than that of mock transfectants in vitro, and lung metastasis after i.v. injection of the transfectants into nude mice was significantly suppressed. These findings indicated that the core structures of O glycans are profoundly involved in the metastatic capacity of cancer cells. PMID- 15755814 TI - Role of Toll-like receptors 2 and 4 in the induction of cyclooxygenase-2 in vascular smooth muscle. AB - Bacteria stimulate macrophages as part of normal host defense. However, when this response is not limited, vascular smooth muscle may also be activated to express "vasoactive" genes (e.g., cyclooxygenase), leading to vascular collapse and septic shock. In macrophages, Toll-like receptors (TLRs) 4 and 2 transduce responses to Gram-negative and Gram-positive bacteria, respectively. However, the role of these TLRs in sensing bacteria in vascular smooth muscle is unclear. To address this question, we have cultured vascular smooth muscle cells from mice deficient in TLR4 (TLR4(-/-) mice), mice deficient in TLR2 (TLR2(-/-) mice), or control mice. Cells cultured from control or TLR2(-/-) mice, but not from TLR4(-/ ) mice, expressed cyclooxygenase-2 and released increasing levels of prostaglandin E(2) after stimulation with whole Escherichia coli bacteria; the combination of IL-1beta plus TNF-alpha induced cyclooxygenase-2 in cells cultured from all three groups of animals. By contrast, Staphylococcus aureus affected cyclooxygenase-2 expression in two distinct ways. First, S. aureus induced a transient inhibition of cyclooxygenase-2 expression, which was overcome with time, and increased protein expression was noted. The effects of S. aureus on cyclooxygenase-2 expression were TLR2- and not TLR4-dependent. Thus, we show that Gram-positive and Gram-negative bacteria induce cyclooxygenase-2 in vascular smooth muscle with differing temporal profiles but with appropriate TLR2-versus TLR4 signaling. These data have important implications for our understanding of the innate immune response in vascular cells and how it may impact vascular disease. PMID- 15755816 TI - Cortisol and ACTH responses to severe asphyxia in preterm fetal sheep. AB - It has been hypothesized that the hypothalamic-pituitary-adrenal (HPA) axis is immature in the preterm fetus and that this compromises their ability to adapt to hypoxic stress; however, there are few direct data. We therefore examined the effects of asphyxia on HPA responses in chronically instrumented preterm fetal sheep (104 days of gestation; term is 147 days), allocated to a sham control group (n = 7) or 25 min of complete umbilical cord occlusion (n = 8), followed by recovery for 72 h. During umbilical cord occlusion there was a rapid rise in ACTH levels (230.4 +/- 63.5 versus 14.1 +/- 1.8 ng ml(-1) in sham controls, 16-fold) and cortisol levels (7.4 +/- 4.9 versus 0.2 +/- 0.1 ng ml(-1), 31-fold), with further increases after release of cord occlusion. ACTH levels were normalized by 24 h, while plasma cortisol levels returned to sham control values 72 h after asphyxia. Fetal arterial blood pressure was elevated in the first 36 h, with a marked increase in femoral vascular resistance, and correlated positively with cortisol levels after asphyxia (P = 0.05). In conclusion, the preterm fetus shows a brisk, substantial HPA response to severe hypoxia. PMID- 15755817 TI - Emphysema-induced reductions in locomotory skeletal muscle contractile function. AB - Patients with COPD suffer from locomotory skeletal muscle contractile dysfunction. This may be due to the disease per se or as a result of some confounding factor. Therefore, the purpose of this investigation was to determine whether emphysema: (1) reduces force production; (2) increases fatigability; and (3) impairs the speed of recovery in locomotory skeletal muscle in an accepted animal model in which many confounding variables can be controlled. To explore this issue, in situ mechanical properties of gastrocnemius were measured in Syrian Golden hamsters 8 months after intratracheal instillation of either saline (control, n = 5) or elastase (emphysema, n = 7). Emphysema increased excised lung volume (80%; P < 0.01), increased fatigability (control, 25% reduction in maximal strength after 4 min of repeated contractions; emphysema, 55% reduction; P < 0.05) and decreased the recovery rate (half-times of recovery: control, 7 +/- 7 s; emphysema, 92 +/- 92 s; P < 0.05) of gastrocnemius muscle. In contrast, emphysema had no effect on maximal force, whether related to body mass or muscle mass, or force-velocity characteristics of gastrocnemius muscle. These data demonstrate that emphysema, independent of physical activity levels, pharmacological intervention, and/or nutritional status, can increase fatigability and impair the speed of recovery of locomotory skeletal muscle contractile function which may contribute to exercise intolerance of COPD patients. PMID- 15755815 TI - Integrons in Xanthomonas: a source of species genome diversity. AB - Integrons are best known for assembling antibiotic resistance genes in clinical bacteria. They capture genes by using integrase-mediated site-specific recombination of mobile gene cassettes. Integrons also occur in the chromosomes of many bacteria, notably beta- and gamma-Proteobacteria. In a survey of Xanthomonas, integrons were found in all 32 strains representing 12 pathovars of two species. Their chromosomal location was downstream from the acid dehydratase gene, ilvD, suggesting that an integron was present at this site in the ancestral xanthomonad. There was considerable sequence and structural diversity among the extant integrons. The majority of integrase genes were predicted to be inactivated by frameshifts, stop codons, or large deletions, suggesting that the associated gene cassettes can no longer be mobilized. In support, groups of strains with the same deletions or stop codons/frameshifts in their integrase gene usually contained identical arrays of gene cassettes. In general, strains within individual pathovars had identical cassettes, and these exhibited no similarity to cassettes detected in other pathovars. The variety and characteristics of contemporary gene cassettes suggests that the ancestral integron had access to a diverse pool of these mobile elements, and that their genes originated outside the Xanthomonas genome. Subsequent inactivation of the integrase gene in particular lineages has largely fixed the gene cassette arrays in particular pathovars during their differentiation and specialization into ecological niches. The acquisition of diverse gene cassettes by different lineages within Xanthomonas has contributed to the species-genome diversity of the genus. The role of gene cassettes in survival on plant surfaces is currently unknown. PMID- 15755818 TI - Antioxidants and ecto-5'-nucleotidase are not involved in the training-induced cardioprotection against ischaemia-reperfusion injury. AB - Isolated Langendorff-perfused hearts from sedentary and prolonged (24 weeks) treadmill-trained rats were subjected to 30 min of normoxic perfusion either alone or followed by 20 min of global ischaemia, or by 20 min of global ischaemia and 15 min of normoxic reperfusion. Pre-ischaemic values of antioxidant enzyme activities and ecto-5'-nucleotidase activity were not different in sedentary and trained hearts but a 5-fold increase of 72-kDa heat shock protein (HSP72) levels was detected in trained myocardium. After ischaemia and reperfusion (I/R), metabolic recovery was better in trained than in sedentary hearts as indicated by higher ATP and creatine phosphate levels. However, antioxidant enzymatic activities, glutathione reductase, and total and mitochondrial superoxide dismutase decreased in trained rats after I/R, whereas they remained unchanged in the sedentary ones. Ecto-5'-nucleotidase activity was modified by I/R in sedentary as well as in trained hearts while HSP72 content did not change. Ecto 5'-nucleotidase activity and HSP72 content increased in parallel by the 30-min perfusion period. In conclusion, the cardioprotection induced by long-term training could be mediated by the exercise-induced increase in HSP72 levels and is not related to enhanced antioxidant systems or ecto-5'-NT activity. PMID- 15755819 TI - Caffeine and arrhythmia. PMID- 15755820 TI - Cocoa, diabetes, and hypertension: should we eat more chocolate? PMID- 15755821 TI - Survival advantages of obesity in dialysis patients. AB - In the general population, a high body mass index (BMI; in kg/m(2)) is associated with increased cardiovascular disease and all-cause mortality. However, the effect of overweight (BMI: 25-30) or obesity (BMI: >30) in patients with chronic kidney disease (CKD) undergoing maintenance hemodialysis (MHD) is paradoxically in the opposite direction; ie, a high BMI is associated with improved survival. Although this "reverse epidemiology" of obesity or dialysis-risk-paradox is relatively consistent in MHD patients, studies in CKD patients undergoing peritoneal dialysis have yielded mixed results. Growing confusion has developed among physicians, some of whom are no longer confident about whether to treat obesity in CKD patients. A similar reverse epidemiology of obesity has been described in geriatric populations and in patients with chronic heart failure (CHF). Possible causes of the reverse epidemiology of obesity include a more stable hemodynamic status, alterations in circulating cytokines, unique neurohormonal constellations, endotoxin-lipoprotein interaction, reverse causation, survival bias, time discrepancies among competitive risk factors, and malnutrition-inflammation complex syndrome. Reverse epidemiology may have significant clinical implications in the management of dialysis, CHF, and geriatric patients, ie, populations with extraordinarily high mortality. Exploring the causes and consequences of the reverse epidemiology of obesity in dialysis patients can enhance our insights into similar paradoxes observed for other conventional risk factors, such as blood pressure and serum cholesterol and homocysteine concentrations, and in other populations such as those with CHF, advanced age, cancer, or AIDS. Weight-gaining interventional studies in dialysis patients are urgently needed to ascertain whether they can improve survival and quality of life. PMID- 15755822 TI - Comparison of abdominal adiposity and overall obesity in predicting risk of type 2 diabetes among men. AB - BACKGROUND: Obesity is a strong risk factor for type 2 diabetes. However, few studies have compared the predictive power of overall obesity with that of central obesity. The cutoffs for waist circumference (WC) and waist-to-hip ratio (WHR) as measures of abdominal adiposity remain controversial. OBJECTIVE: The objective was to compare body mass index (BMI), WC, and WHR in predicting type 2 diabetes. DESIGN: A prospective cohort study (Health Professionals Follow-Up Study) of 27 270 men was conducted. WC, WHR, and BMI were assessed at baseline. Covariates and potential confounders were assessed repeatedly during the follow up. RESULTS: During 13 y of follow-up, we documented 884 incident type 2 diabetes cases. Age-adjusted relative risks (RRs) across quintiles of WC were 1.0, 2.0, 2.7, 5.0, and 12.0; those of WHR were 1.0, 2.1, 2.7, 3.6, and 6.9; and those of BMI were 1.0, 1.1, 1.8, 2.9, and 7.9 (P for trend < 0.0001 for all). Multivariate adjustment for diabetes risk factors only slightly attenuated these RRs. Adjustment for BMI substantially attenuated RRs for both WC and WHR. The receiver operator characteristic curve analysis indicated that WC and BMI were similar and were better than WHR in predicting type 2 diabetes. The cumulative proportions of type 2 diabetes cases identified according to medians of BMI (>/=24.8), WC (>/=94 cm), and WHR (>/=0.94) were 82.5%, 83.6%, and 74.1%, respectively. The corresponding proportions were 78.9%, 50.5%, and 65.7% according to the recommended cutoffs. CONCLUSIONS: Both overall and abdominal adiposity strongly and independently predict risk of type 2 diabetes. WC is a better predictor than is WHR. The currently recommended cutoff for WC of 102 cm for men may need to be reevaluated; a lower cutoff may be more appropriate. PMID- 15755823 TI - Genetic and environmental influences on eating behavior: the Swedish Young Male Twins Study. AB - BACKGROUND: Eating behavior may be implicated in the increasing prevalence of overweight and obesity, presumably in relation to easy access to energy-dense and highly palatable foods. OBJECTIVE: The aim of the present study was to disentangle genetic and environmental influences on eating behavior in a population-based cohort of male twins. DESIGN: The study included 326 dizygotic and 456 monozygotic male twin pairs aged 23-29 y from Sweden. The revised 21-item version of the Three-Factor Eating Questionnaire (TFEQ-R21) was used to assess eating behavior. This validated instrument consists of 3 dimensions: cognitive restraint, emotional eating, and uncontrolled eating. Structural equation modeling was used to estimate the heritability of eating behavior. RESULTS: Cognitive restraint was the only TFEQ-R21 scale that significantly correlated with BMI (r = 0.39, P < 0.0001). The best-fitted models gave a heritability of 59% (95% CI: 52%, 66%) for cognitive restraint, 60% (95% CI: 52%, 67%) for emotional eating, and 45% (95% CI: 36%, 53%) for uncontrolled eating. CONCLUSIONS: These results show the great importance of genetic factors in the eating behavior of a large, unselected population of young adult male twins. Nonshared environmental factors were also important, whereas shared environmental factors did not contribute to eating behavior. PMID- 15755824 TI - Decreases in fasting leptin and insulin concentrations after acute energy restriction and subsequent compensation in food intake. AB - BACKGROUND: The decrease in leptin after energy restriction is a starvation signal to the brain. Several studies have found an association between this decrease and subjective appetite; however, no solid data are available on the acute decrease in fasting leptin concentration and subsequent caloric compensation. OBJECTIVE: The objective was to assess the effect of acute decreases in fasting leptin concentrations, induced by energy restriction, on subsequent energy intake compensation. We hypothesized that men with a large decrease in fasting leptin concentrations would have larger ad libitum energy intakes than would men with a small decrease in leptin. DESIGN: Thirty-four male unrestrained eaters [age: 23 +/- 3 y; body mass index (in kg/m(2)): 22.3 +/- 1.6] participated in a semicontrolled intervention study. Fasting serum leptin and insulin concentrations were measured before and 2 d after 62% energy restriction. Energy intake was measured on the 2 following days on which food was provided ad libitum. RESULTS: During energy restriction, fasting leptin and insulin concentrations decreased by 27.2% (95% CI: -34.4%, -19.9%) and 30.7% (95% CI: 41.0%, -20.4%), respectively. Subjects consumed 143 +/- 27% of their estimated energy requirements (18.3 +/- 2.9 MJ) on the first day and 124 +/- 20% (16.0 +/- 2.6 MJ) on the second day of ad libitum intake. No significant correlations were observed between decreases in fasting leptin or insulin concentrations and subsequent ad libitum energy intake; however, decreases in insulin were correlated with an increase in carbohydrate intake (r=-0.49, P < 0.01). CONCLUSION: Although fasting leptin concentrations decreased significantly during energy restriction and subjects showed compensatory behavior during subsequent ad libitum food intake, no association was observed between the decrease in fasting leptin concentrations and caloric compensation. PMID- 15755825 TI - Caffeine and risk of atrial fibrillation or flutter: the Danish Diet, Cancer, and Health Study. AB - BACKGROUND: It is not known whether the consumption of caffeine is associated with excess risk of atrial fibrillation. OBJECTIVE: We evaluated the risk of atrial fibrillation or flutter in association with daily consumption of caffeine from coffee, tea, cola, cocoa, and chocolate. DESIGN: We prospectively examined the association between the amount of caffeine consumed per day and the risk of atrial fibrillation or flutter among 47 949 participants (x age: 56 y) in the Danish Diet, Cancer, and Health Study. Subjects were followed in the Danish National Registry of Patients and in the Danish Civil Registration System. The consumption of caffeine was analyzed by quintiles with Cox proportional-hazard models. RESULTS: During follow-up (x: 5.7 y), atrial fibrillation or flutter developed in 555 subjects (373 men and 182 women). When the lowest quintile of caffeine consumption was used as a reference, the adjusted hazard ratios (95% CIs) in quintiles 2, 3, 4, and 5 were 1.12 (0.87, 1.44), 0.85 (0.65, 1.12), 0.92 (0.71, 1.20), and 0.91 (0.70, 1.19), respectively. CONCLUSION: Consumption of caffeine was not associated with risk of atrial fibrillation or flutter. PMID- 15755827 TI - Blood lipid and oxidative stress responses to soy protein with isoflavones and phytic acid in postmenopausal women. AB - BACKGROUND: Postmenopausal women are at risk of cardiovascular disease (CVD) as a result of unfavorable blood lipid profiles and increased oxidative stress. Soy protein consumption may help protect against these risk factors. OBJECTIVE: Our objective was to ascertain the effect of the soy protein components isoflavones and phytate on CVD risk in postmenopausal women. DESIGN: In a double-blind 6-wk study, 55 postmenopausal women were randomly assigned to 1 of 4 treatments with soy protein (40 g/d) isolate (SPI): low phytate/low isoflavone (LP/LI); normal phytate/low isoflavone (NP/LI); low phytate/normal isoflavone (LP/NI); or normal phytate/normal isoflavone (NP/NI). Blood lipids (total, LDL, and HDL cholesterol and triacylglycerol) and oxidative stress indexes (protein carbonyls, oxidized LDLs, and 8-iso-prostaglandin-F(2alpha)) were measured at baseline and 6 wk. RESULTS: The oxidative stress indexes were not significantly affected by either phytate or isoflavones. Phytate treatment had a minimal but nonsignificant effect in reducing protein carbonyls and 8-iso-prostaglandin-F(2alpha); the reductions were 6-8% and 4-6% in the NP/LI and NP/NI groups and 1-4% and 3-4% in the LP/LI and LP/NI groups, respectively. Similarly, circulating lipids were not significantly affected by either phytate or isoflavones. The decline in total (6% 7% compared with 2%-4%) and LDL (10%-11% compared with 3%-7%) cholesterol did not differ significantly between the normal- and low-isoflavone groups, respectively. CONCLUSION: In postmenopausal women, neither phytate nor isoflavones in SPI have a significant effect of reducing oxidative damage or favorably altering blood lipids. PMID- 15755826 TI - Influence of long-term intervention with dietary counseling, long-chain n-3 fatty acid supplements, or both on circulating markers of endothelial activation in men with long-standing hyperlipidemia. AB - BACKGROUND: Dietary factors and very-long-chain n-3 polyunsaturated fatty acids (n-3 PUFAs) may influence the atherothrombotic process. Elevated concentrations of circulating cell adhesion molecules, thrombomodulin (TM), von Willebrand factor (vWF), and tissue-type plasminogen activator antigen (tPAag) are related to atherothrombotic cardiovascular disease. OBJECTIVE: The randomized Diet and Omega-3 Intervention Trial (DOIT) targeted a comparison of the effect of 3-y dietary counseling, n-3 PUFA supplementation (2.4 g/d), or both on circulating markers of endothelial activation. DESIGN: The study included 563 elderly men with long-standing hyperlipidemia. The men were randomly assigned by factorial design into 4 groups: control (no dietary counseling and placebo capsules), dietary counseling (and placebo capsules), n-3 PUFA supplementation (no dietary counseling), and dietary counseling and n-3 PUFA supplementation. RESULTS: Serum concentrations of fatty acids reflected good compliance. Dietary counseling was followed by significantly reduced concentrations of soluble intercellular adhesion molecule 1 (sICAM-1; P < 0.001), sTM (P = 0.004), and tPAag (P < 0.001) than in subjects without dietary counseling. After n-3 PUFA supplementation, significantly reduced concentrations of sICAM-1 (P < 0.001) and sTM (P = 0.006) were observed when compared with subjects receiving placebo capsules. An increase in tPAag was not significantly different from that observed in subjects receiving placebo capsules. For sICAM-1, a significant effect was observed for both interventions combined. CONCLUSIONS: Each intervention (dietary counseling or n-3 PUFA supplements) reduced sTM and sICAM-1 concentrations, indicating decreased endothelial activation. The tPAag increase in the groups not receiving dietary counseling (pooled), which indicates progression of atherosclerosis, was significantly counteracted by dietary counseling. PMID- 15755828 TI - Validity of impedance-based equations for the prediction of total body water as measured by deuterium dilution in African women. AB - BACKGROUND: Little information is available on the validity of simple and indirect body-composition methods in non-Western populations. Equations for predicting body composition are population-specific, and body composition differs between blacks and whites. OBJECTIVE: We tested the hypothesis that the validity of equations for predicting total body water (TBW) from bioelectrical impedance analysis measurements is likely to depend on the racial background of the group from which the equations were derived. DESIGN: The hypothesis was tested by comparing, in 36 African women, TBW values measured by deuterium dilution with those predicted by 23 equations developed in white, African American, or African subjects. These cross-validations in our African sample were also compared, whenever possible, with results from other studies in black subjects. RESULTS: Errors in predicting TBW showed acceptable values (1.3-1.9 kg) in all cases, whereas a large range of bias (0.2-6.1 kg) was observed independently of the ethnic origin of the sample from which the equations were derived. Three equations (2 from whites and 1 from blacks) showed nonsignificant bias and could be used in Africans. In all other cases, we observed either an overestimation or underestimation of TBW with variable bias values, regardless of racial background, yielding no clear trend for validity as a function of ethnic origin. CONCLUSIONS: The findings of this cross-validation study emphasize the need for further fundamental research to explore the causes of the poor validity of TBW prediction equations across populations rather than the need to develop new prediction equations for use in Africa. PMID- 15755829 TI - Short-term protein intake and stimulation of protein synthesis in stunted children with cystic fibrosis. AB - BACKGROUND: Stunted children with cystic fibrosis (CF) have less net protein anabolism than do children without CF, and the result is retarded growth in the CF patients. It is not known whether protein intake above that recommended by the Cystic Fibrosis Foundation would further stimulate whole-body protein synthesis. OBJECTIVE: We studied the effects of 3 amounts of protein intake on whole-body protein synthesis and breakdown by using isotopic infusion of [1-(13)C]valine and [(15)N(2)]urea in children with stable CF who required tube feeding. DESIGN: In 8 pediatric CF patients, we administered 3 randomly allocated isocaloric diets with normal (NP), intermediate (IP), and high (HP) amounts of protein (1.5, 3, and 5 g . kg(-1) . d(-1), respectively) by continuous drip feeding during a 4-d period at 6-wk intervals. Each patient acted as his or her own control. On the fourth day of feeding, whole-body protein synthesis and breakdown were measured. RESULTS: Protein synthesis was significantly higher in the HP group (x +/- SEM: 1.78 +/- 0.07 micromol . kg(-1) . min(-1)) than in the IP (1.57 +/- 0.08 micromol . kg(-1) . min(-1); P=0.001) and NP (1.37 +/- 0.07 micromol . kg(-1) . min(-1); P < 0.001) groups. There were no significant differences in protein breakdown. Net retention of nitrogen was significantly higher in the HP group (12.93 +/- 1.42 micromol . kg(-1) . min(-1)) than in the IP (7.61 +/- 1.40 micromol . kg(-1) . min(-1); P=0.01) and HP (2.48 +/- 0.20 micromol . kg(-1) . min(-1); P < 0.001) groups. CONCLUSION: In stunted children with CF requiring tube feeding, the highest stimulation of whole-body protein synthesis was achieved with a short-term dietary protein intake of 5 g . kg(-1) . d(-1). PMID- 15755830 TI - Short-term administration of dark chocolate is followed by a significant increase in insulin sensitivity and a decrease in blood pressure in healthy persons. AB - BACKGROUND: Numerous studies indicate that flavanols may exert significant vascular protection because of their antioxidant properties and increased nitric oxide bioavailability. In turn, nitric oxide bioavailability deeply influences insulin-stimulated glucose uptake and vascular tone. Thus, flavanols may also exert positive metabolic and pressor effects. OBJECTIVE: The objective was to compare the effects of either dark or white chocolate bars on blood pressure and glucose and insulin responses to an oral-glucose-tolerance test in healthy subjects. DESIGN: After a 7-d cocoa-free run-in phase, 15 healthy subjects were randomly assigned to receive for 15 d either 100 g dark chocolate bars, which contained approximately 500 mg polyphenols, or 90 g white chocolate bars, which presumably contained no polyphenols. Successively, subjects entered a further cocoa-free washout phase of 7 d and then were crossed over to the other condition. Oral-glucose-tolerance tests were performed at the end of each period to calculate the homeostasis model assessment of insulin resistance (HOMA-IR) and the quantitative insulin sensitivity check index (QUICKI); blood pressure was measured daily. RESULTS: HOMA-IR was significantly lower after dark than after white chocolate ingestion (0.94 +/- 0.42 compared with 1.72 +/- 0.62; P < 0.001), and QUICKI was significantly higher after dark than after white chocolate ingestion (0.398 +/- 0.039 compared with 0356 +/- 0.023; P = 0.001). Although within normal values, systolic blood pressure was lower after dark than after white chocolate ingestion (107.5 +/- 8.6 compared with 113.9 +/- 8.4 mm Hg; P < 0.05). CONCLUSION: Dark, but not white, chocolate decreases blood pressure and improves insulin sensitivity in healthy persons. PMID- 15755831 TI - Serum transferrin receptor and zinc protoporphyrin as indicators of iron status in African children. AB - BACKGROUND: Although transferrin receptor (TfR) and zinc protoporphyrin (ZnPP) are often used to define iron status in school-age children in developing countries, the diagnostic cutoffs for this age group are uncertain. OBJECTIVE: The objective was to determine the sensitivity and specificity of TfR and ZnPP in predicting iron deficiency in black and white children in Africa. DESIGN: Hemoglobin, C-reactive protein (CRP), serum ferritin (SF), TfR, and ZnPP were measured in children in Cote d'Ivoire and Morocco. We excluded children with elevated CRP and then used receiver operating characteristic (ROC) curves to evaluate TfR and ZnPP alone and in combination in screening for iron deficiency, defined as an SF concentration <15 mug/L, and iron deficiency anemia (IDA), defined as an SF concentration <15 mug/L and low hemoglobin. RESULTS: The sample included 2814 children aged 5-15 y. The sensitivity and specificity of TfR and ZnPP were limited by considerable overlap between iron-sufficient, nonanemic children and those with IDA. On the basis of ROC curves, we identified diagnostic cutoffs for TfR and ZnPP that achieved specificities and sensitivities of approximately 60-80%. Separate cutoffs for Cote d'Ivoire and Morocco gave the best performance; the cutoffs for both TfR and ZnPP were higher in Cote d'Ivoire. Moreover, a comparison of nonanemic, iron-sufficient subjects showed that Ivorian children had significantly higher TfR and ZnPP concentrations than did Moroccan children (P < 0.01). CONCLUSIONS: New diagnostic cutoffs for TfR and ZnPP, based on ROC curve analyses, may improve the performance of these indexes in defining iron status in children. Significant ethnic differences in TfR and ZnPP suggest that separate cutoffs may be needed for black and white children. PMID- 15755832 TI - Influence of apolipoprotein E genotype on fat-soluble plasma antioxidants in Spanish children. AB - BACKGROUND: Apolipoprotein (apo) E is a major determinant of plasma lipid concentrations, which in turn influence the plasma concentrations of various fat soluble vitamins. OBJECTIVE: We aimed to analyze the effect of APOE genotype on fat-soluble antioxidant concentrations in children. DESIGN: A total of 926 healthy boys and girls aged 6-8 y were selected from 4 cities in Spain. APOE genotyping was carried out, and plasma concentrations of lipids, apolipoproteins, and lipid-soluble antioxidants were measured. RESULTS: Plasma lipid concentrations were strongly influenced by APOE genotype. The mean plasma concentration of alpha-tocopherol was 21.3 micromol/L, which is one of the highest values ever reported for a population of children. Although plasma concentrations of alpha-tocopherol, gamma-tocopherol, lycopene, and alpha carotene varied significantly between subjects with different APOE genotypes, most of these differences disappeared after adjustment for lipoprotein-related covariates. Nevertheless, tocopherol concentrations remained elevated in individuals with the E2/2 genotype. Multivariate regression analysis showed interactions of APOE genotype with triacylglycerol and apo B in determining alpha tocopherol concentrations. When subjects were stratified according to major apo E groups, apo B appeared to be the most important predictor of alpha-tocopherol concentrations in all groups, whereas triacylglycerol was identified only in carriers of the E2 allele. CONCLUSIONS: The association between APOE genotype and lipophilic antioxidant concentrations is dependent mainly on the effect of the polymorphism on lipoprotein concentrations. However, triacylglycerol plays a role in determining the variability of alpha-tocopherol concentrations in E2 carriers only. This suggests that the alpha-tocopherol content in each lipoprotein class varies according to APOE genotype. PMID- 15755833 TI - Concentrations of retinoids in early pregnancy and in newborns and their mothers. AB - BACKGROUND: Retinoids are vital for embryonic development; both excesses and deficiencies of vitamin A are known to give similar patterns of birth defects. Concentrations of retinol in newborns and in pregnant women have been investigated, but concentrations of the biologically active metabolite all-trans retinoic acid and its isomer 13-cis retinoic acid have not. OBJECTIVE: We measured serum concentrations of these retinoid derivatives in newborns and their mothers and in women in the first trimester of pregnancy, when embryonic differentiation (organogenesis) takes place. DESIGN: In this descriptive study, 10 newborns from normal deliveries and their mothers and 16 healthy women in their first trimester of pregnancy were studied. Seventeen healthy women served as control subjects. all-trans and 13-cis Retinoic acid and retinol concentrations were measured by HPLC. RESULTS: The newborns had significantly lower retinol concentrations (1.0 micromol/L) than did their mothers (1.7 micromol/L; P = 0.013). Serum all-trans retinoic acid was also significantly lower in the newborns (3.4 nmol/L) than in their mothers (5.8 nmol/L; P = 0.008). In addition, serum concentrations of 13-cis retinoic acid were significantly lower in the newborns (2.0 nmol/L) than in their mothers (2.6 nmol/L; P = 0.005). The serum concentrations of all-trans retinoic acid and retinol did not correlate in any group. CONCLUSION: Retinol concentrations do not accurately reflect the concentrations of the biologically active derivative all-trans retinoic acid. PMID- 15755834 TI - Longitudinal measurements of zinc absorption in Peruvian children consuming wheat products fortified with iron only or iron and 1 of 2 amounts of zinc. AB - BACKGROUND: Information is needed on the fractional absorption of zinc (FAZ) and absorbed zinc (AZ) during prolonged exposure to zinc-fortified foods. OBJECTIVE: The objective was to measure FAZ and AZ from diets fortified with different amounts of zinc and to determine whether zinc absorption changes over approximately 7 wk. DESIGN: Forty-one stunted, moderately anemic children received daily, at breakfast and lunch, 100 g wheat products fortified with 3 mg Fe (ferrous sulfate) and 0 (group Zn-0), 3 (group Zn-3), or 9 (group Zn-9) mg Zn (zinc sulfate) per 100 g flour. FAZ was measured on days 2-3 and 51-52; meal specific AZs were calculated as the product of FAZ and zinc intake. RESULTS: For the breakfast and lunch meals combined, mean total zinc intakes were 2.14, 4.72, and 10.04 mg/d in groups Zn-0, Zn-3, and Zn-9, respectively, during the initial absorption studies; mean (+/-SD) FAZ values were 0.341 +/- 0.111, 0.237 +/- 0.052, and 0.133 +/- 0.041, respectively, on days 2-3 (P < 0.001) and did not change significantly on days 51-52 in the subset of 31 children studied twice. Mean initial AZ was positively related to zinc intake (0.71 +/- 0.18, 1.11 +/- 0.21, and 1.34 +/- 0.47 mg/d, respectively; P < 0.001); final values did not differ significantly from the initial values. CONCLUSIONS: AZ from meals containing zinc-fortified wheat products increases in young children relative to the level of fortification and changes only slightly during approximately 7-wk periods of consumption. Although consumption of zinc-fortified foods may reduce FAZ, zinc fortification at the levels studied positively affects total daily zinc absorption, even after nearly 2 mo of exposure to zinc-fortified diets. PMID- 15755835 TI - Dietary vitamin B-6 restriction does not alter rates of homocysteine remethylation or synthesis in healthy young women and men. AB - BACKGROUND: The effects of vitamin B-6 status on steady-state kinetics of homocysteine metabolism in humans are unclear. OBJECTIVE: The objective was to determine the effects of dietary vitamin B-6 restriction on the rates of homocysteine remethylation and synthesis in healthy humans. DESIGN: Primed, constant infusions of [(13)C(5)]methionine, [3-(13)C]serine, and [(2)H(3)]leucine were conducted in healthy female (n=5) and male (n=4) volunteers (20-30 y) before and after 4 wk of dietary vitamin B-6 restriction (<0.5 mg vitamin B-6/d) to establish whether vitamin B-6 status affects steady-state kinetics of homocysteine metabolism in the absence of concurrent methionine intake. Effects of dietary vitamin B-6 restriction on vitamin B-6 status, plasma amino acid concentrations, and the rates of reactions of homocysteine metabolism were assessed. RESULTS: Dietary vitamin B-6 restriction significantly reduced plasma pyridoxal 5-phosphate (PLP) concentrations (55.1 +/- 8.3 compared with 22.6 +/- 1.3 nmol/L; P=0.004), significantly increased plasma glycine concentrations (230 +/- 14 compared with 296 +/- 15; P=0.008), and significantly reduced basal (43%; P < 0.001) and PLP-stimulated (35%; P=0.004) lymphocyte serine hydroxymethyltransferase activities in vitro. However, the in vivo fluxes of leucine, methionine, and serine; the rates of homocysteine synthesis and remethylation (total and vitamin B-6-dependent); and the concentrations of homocysteine, methionine, and serine in plasma were not significantly affected by dietary vitamin B-6 restriction. CONCLUSIONS: Moderate vitamin B-6 deficiency does not significantly alter the rates of homocysteine remethylation or synthesis in healthy young adults in the absence of dietary methionine intake. PMID- 15755836 TI - Changes in iodine excretion in 50-69-y-old denizens of an Arctic society in transition and iodine excretion as a biomarker of the frequency of consumption of traditional Inuit foods. AB - BACKGROUND: Iodine intake in Greenland has been hypothesized to exceed 10 times the recommended amount. The transition from a traditional Arctic society may change the iodine intake, but no field studies have been performed. OBJECTIVE: We aimed to ascertain iodine intakes, factors affecting iodine intake in circumpolar populations, and the usefulness of urinary iodine excretion as a biomarker for validation of Inuit food-frequency questionnaires. DESIGN: Data were collected in a cohort study of 4 Greenland population groups: Inuit living in the capital city, the major town, and settlements in East Greenland and non-Inuit. Supplement use and lifestyle factors were evaluated with questionnaires, and dietary habits were ascertained with a food-frequency questionnaire. Iodine was measured in spot urine samples. RESULTS: One percent of the population of Greenland was invited, and the participation rate was 95%. Less than 5% of Inuit but 55% of non-Inuit had urinary iodine excretion < 50 microg/24 h. Median urinary iodine excretion declined with the degree of decrease in the traditional lifestyle: it was 198, 195, 147, and 58 microg/24 h among Inuit in settlements, town, and city and in non-Inuit, respectively (P < 0.001). Participants were divided into diet groups calculated from Inuit food frequency. Iodine excretion decreased with increasing intake of imported foods (P < 0.001). In regression models, type of diet and the subject's lifestyle, sex, weight, ethnicity, and intake of iodine-containing supplements affected urinary iodine excretion. CONCLUSIONS: Circumpolar non-Inuit are at risk of iodine deficiency. Departure from the traditional Inuit diet lowers iodine intake, which should be monitored in Arctic societies. Urinary iodine excretion may be a useful biomarker of traditional Inuit food frequency. PMID- 15755837 TI - Common dihydrofolate reductase 19-base pair deletion allele: a novel risk factor for preterm delivery. AB - BACKGROUND: Folate is critical for cell division, a major feature of in utero development. Dihydrofolate reductase (DHFR) is required to convert the folic acid used in supplements and for food fortification and the dihydrofolate produced by thymidylate synthase during DNA synthesis to the reduced folate forms used by the cell. OBJECTIVE: We aimed to determine whether a common, recently discovered deletion polymorphism in the DHFR gene is a risk factor for preterm delivery or low birth weight. DESIGN: We studied 324 pregnant women from Camden, NJ. Folate intake was computed from folate supplement intake plus the mean of two 24-h recalls completed during the course of pregnancy. Genomic DNA was extracted from the women's leukocytes and genotyped. RESULTS: Women with a deletion allele had a significantly greater risk of preterm delivery [adjusted odds ratio (AOR): 3.0; 95% CI: 1.0, 8.8; P < 0.05] than did those without a deletion allele. Women with both a DHFR deletion allele and low folate intake (<400 microg/d from diet plus supplements) had a significantly greater risk of preterm delivery (AOR: 5.5; 95% CI: 1.5, 20.4; P = 0.01) and a significantly greater risk of having an infant with a low birth weight (AOR: 8.3; 95% CI: 1.8, 38.6; P = 0.01) than did women without a deletion allele and with a folate intake >/=400 microg/d. CONCLUSIONS: The DHFR 19-base pair deletion allele may be a risk factor for preterm delivery. In the presence of low dietary folate, the allele may also be a risk factor for low birth weight. This may be a gene-environment interaction. PMID- 15755838 TI - Status of plasma folate after folic acid fortification of the food supply in pregnant African American women and the influences of diet, smoking, and alcohol consumption. AB - BACKGROUND: African American women and socioeconomically challenged women are at risk of compromised folate status and, thus, of folate-related birth defects. Data are limited on circulating folate concentrations in pregnant African American women after folic acid fortification of the food supply was implemented. OBJECTIVE: The objective was to determine the influence of smoking and alcohol consumption on plasma 5-methyltetrahydrofolic acid (5-MTHFA) concentrations in pregnant African American women. DESIGN: Alcohol consumption, smoking exposure, and other characteristics of pregnant African American women reporting to an inner-city antenatal clinic were assessed. At 24 wk of gestation, blood samples and food-frequency intake data were collected. Plasma 5-MTHFA concentrations were determined by liquid chromatography-mass spectrometry for 116 subjects and examined in a correlational study design. RESULTS: Dietary folate and markers of alcohol consumption were positively associated, whereas exposure to smoke was negatively associated with plasma 5-MTHFA. More than one-half of the participants in this population failed to meet the recommended dietary allowance for dietary folate equivalents of 600 microg/d during pregnancy. CONCLUSIONS: Most inner-city African American women are not meeting the recommended dietary allowance for dietary folate during pregnancy, and smoking may further compromise their folate status. Programs to reduce smoking and raise awareness about the importance of folate and multivitamin supplementation during pregnancy need to target this population. PMID- 15755839 TI - Changes in basal metabolic rate during pregnancy in relation to changes in body weight and composition, cardiac output, insulin-like growth factor I, and thyroid hormones and in relation to fetal growth. AB - BACKGROUND: The total energy cost of pregnancy is largely due to an elevated basal metabolic rate (BMR). Large variations in the BMR response to pregnancy have been reported, but the factors associated with this variability are incompletely known. OBJECTIVE: The objective was to identify factors associated with variability in the BMR response to pregnancy. DESIGN: In 22 healthy women, BMR, body weight (BW), total body fat (TBF), fat-free mass (FFM), circulatory variables, serum concentrations of insulin-like growth factor I (IGF-I), and thyroid hormones were measured before pregnancy and in gestational weeks 14 and 32. BMR and BW were also measured in gestational weeks 8, 20, and 35. Fetal weight was estimated in gestational week 31. RESULTS: In gestational week 14, the increase in BMR correlated significantly with the corresponding increase in BW and with the prepregnancy percentage of TBF. Together these variables explained approximately 40% of the variability in the BMR response. In gestational week 32, the increase in BMR correlated significantly with the corresponding changes in BW, TBF, FFM, IGF-I, cardiac output, and free triiodothyronine. The increase in BW in combination with fetal weight or with the elevated concentration of IGF-I in serum explained approximately 60% of the variability in the increase in BMR. CONCLUSIONS: Weight gain and the prepregnancy percentage of TBF-ie, factors related to the maternal nutritional situation-are important factors with regard to the variability in the BMR response to pregnancy. Thus, it is important to consider the nutritional situation before and during gestation when assessing pregnancy energy requirements. PMID- 15755840 TI - Phosphatidylcholine and lysophosphatidylcholine excretion is increased in children with cystic fibrosis and is associated with plasma homocysteine, S adenosylhomocysteine, and S-adenosylmethionine. AB - BACKGROUND: Hepatic steatosis and fat malabsorption are common in cystic fibrosis (CF). Choline deficiency results in decreased phosphatidylcholine synthesis through the cytidine diphosphocholine-choline pathway and hepatic steatosis and in increased synthesis of phosphatidylcholine from phosphatidylethanolamine using methyl groups from S-adenosylmethionine. The intestinal absorption of phosphatidylcholine in CF is unknown. OBJECTIVES: The objective was to determine whether excretion of choline phosphoglyceride (phosphatidylcholine and lysophosphatidylcholine) is increased in CF and whether loss of fecal choline phosphoglyceride is associated with altered plasma methionine cycle metabolites. DESIGN: A cross-sectional study involved 53 children with CF and 18 control children without CF. Blood was collected from all participants. A subset of 18 children with CF and 8 control children provided 72-h fecal samples and 5-d food records. RESULTS: Fat absorption was significantly lower (x+/- SEM: 86.2 +/- 1.6% and 94.1 +/- 1.2%) and excretion of fecal fat (12.9 +/- 1.7 and 3.9 +/- 0.7 g/d), phospholipid (median: 130 and 47.7 mg/d), phosphatidylcholine (19.6 and 2.1 mg/d), and lysophosphatidylcholine (60.3 and 16.9 mg/d) was significantly higher in children with CF than in control children, respectively (P < 0.05). Choline phosphoglyceride excretion was positively correlated with plasma homocysteine and S-adenosylhomocysteine and inversely related with plasma methionine (P < 0.05). CONCLUSIONS: Choline phosphoglyceride excretion is increased in children with CF and is associated with decreased plasma methionine and increased homocysteine and S-adenosylhomocysteine. These findings suggest choline depletion and an increased choline synthesis by S-adenosylmethionine-dependent methylation in CF, as well as a metabolic link between phosphatidylcholine metabolism and the methionine homocysteine cycle in humans. PMID- 15755841 TI - Absorption and transport of dietary long-chain fatty acids in cirrhosis: a stable isotope-tracing study. AB - BACKGROUND: In rats, 30-70% of dietary fatty acids (FAs) are absorbed through the portal vein. Whether this occurs in humans is unknown, but it may occur in persons with cirrhosis, who show a blunted chylomicronemic response to dietary fat without significant steatorrhea. OBJECTIVE: The objective was to investigate whether portal FA absorption occurs in humans with cirrhosis. DESIGN: Six control subjects and 10 patients with (n = 5) and without (n = 5) cirrhotic ascites were fed [1-(13)C]palmitic and oleic acids in a test meal. Samples were drawn before and 30, 60, 90, 120, 240, 360, 480, and 720 min afterward for plasma [1-(13)C] labeled FAs and breath (13)CO(2) assay. Fecal [1-(13)C]-labeled FAs were also measured. RESULTS: [1-(13)C]-Labeled FAs increased in chylomicrons in all groups, but less in ascitic cirrhotic patients, because their median area under the curve from 120 to 720 min was significantly lower than in the control subjects for labeled palmitate [520 (interquartile range: 192-1137) compared with 2862 (2674 4175) micromol . min/L] and oleate [829 (781-1263) compared with 3119 (2939-4986) micromol . min/L]. [1-(13)C]-Labeled FA enrichment of VLDL was also lower in cirrhotic patients. [1-(13)C]-Labeled FA in free FAs peaked earlier in ascitic than in nonascitic patients and control subjects, mainly for [1-(13)C]oleate, and the median area under the curve from 0 to 120 min was significantly higher in ascitic patients than in control subjects [301 (255-400) compared with 48 (34 185) micromol . min/L]. Fecal excretion of [1-(13)C]-labeled FA was negligible and not significantly different between groups. CONCLUSIONS: The low [1-(13)C] labeled FA concentrations in chylomicrons and VLDL, without increased fecal losses, confirm previous data in cirrhotic patients with the use of an unlabeled fat load. The earlier [1-(13)C]-labeled FA appearance in free FAs supports the portal absorption of dietary fat in patients with advanced cirrhosis with spontaneous portal-systemic shunting. PMID- 15755842 TI - Investigating heterogeneity in studies of resting energy expenditure in persons with HIV/AIDS: a meta-analysis. AB - BACKGROUND: There is conflict in the literature about the extent of alterations of resting energy expenditure (REE) in persons with HIV. OBJECTIVE: The study was conducted to ascertain the mean difference in REE (in kJ) per kilogram of fat free mass (FFM; REE/FFM) between HIV-positive subjects and control subjects and to investigate heterogeneity in the literature. DESIGN: A meta-analysis comparing classical and Bayesian methods was conducted. Heterogeneity was investigated by using subgroup analysis, metaregression, and a mixed indirect comparison. RESULTS: Of 58 studies meeting the inclusion criteria, 32 included both HIV positive and control groups; 24 of these 32 were included. Thirty-seven studies were used in the mixed indirect comparison, and 30 were used in the subgroup comparisons of the HIV-symptomatic, lipodystrophy, weight-losing, and weight stable subgroups and the healthy (HIV-negative) control group. Mean REE/FFM was significantly higher in 732 HIV-positive subjects than in 340 control subjects [11.93 kJ/kg (95% CI: 8.44,15.43 kJ/kg) and 12.47 kJ/kg (95% CI: 8.19,16.57 kJ/kg), classical and Bayesian random effects, respectively]; the test for heterogeneity was significant (P < 0.001). Both the mixed indirect comparison and the subgroup analysis indicated that REE/FFM was highest in the symptomatic subgroup; however, the small number of studies investigating symptomatic subjects limited statistical comparisons. The presence of lipodystrophy, use of highly active antiretroviral therapy, subject age, and method of body-composition measurement could not explain the heterogeneity in the data with the use of metaregression. CONCLUSIONS: REE/FFM (kJ/kg) is significantly higher in HIV positive subjects than in healthy control subjects. Symptomatic HIV infection may contribute to the variations reported in the literature. PMID- 15755843 TI - Overweight exceeds underweight among women in most developing countries. AB - BACKGROUND: It is generally believed that overweight is less prevalent than undernutrition in the developing world, particularly in rural areas, and that it is concentrated in higher socioeconomic status (SES) groups. OBJECTIVE: The purpose of this study was to examine patterns of adult female overweight and underweight in the developing world by using categories of urban or rural status and SES strata. DESIGN: Body mass index (BMI; in kg/m(2)) data collected in 36 countries from 1992 to 2000 by nationally representative cross-sectional surveys of women aged 20-49 y (n = 148579) were classified as indicating underweight (BMI < 18.5) and overweight (BMI >/= 25). Associations between the nutritional status of urban and rural women and each country's per capita gross national income (GNI) and level of urbanization were explored in the overall sample and among different SES groups. RESULTS: Overweight exceeded underweight in well over half of the countries: the median ratio of overweight to underweight was 5.8 in urban and 2.1 in rural areas. Countries with high GNIs and high levels of urbanization had not only high absolute prevalences of overweight but also small urban-rural differences in overweight and very high ratios of overweight to underweight. In the more-developed countries, overweight among low-SES women was high in both rural (38%) and urban (51%) settings. Even many poor countries, countries in which underweight persists as a significant problem, had fairly high prevalences of rural overweight. CONCLUSIONS: In most developing economies, prevalences of overweight in young women residing in both urban and rural areas are higher than those in underweight women, especially in countries at higher levels of socioeconomic development. Research is needed to assess male and child overweight to understand the dynamics facing these groups as well. PMID- 15755844 TI - Glycemic index and body weight. PMID- 15755845 TI - More support for dietary patterns involving high-fiber, high-complex carbohydrates. PMID- 15755846 TI - Low plasma pyridoxal-5'phosphate and cardiovascular disease risk in women: results from the Coronary Risk Factors for Atherosclerosis in Women Study. PMID- 15755847 TI - The effect of thyroxine treatment started in the neonatal period on development and growth of two-year-old Down syndrome children: a randomized clinical trial. AB - CONTEXT: Young Down syndrome children appear to have a mild form of congenital hypothyroidism that is rarely detected by neonatal screening and usually left untreated. OBJECTIVE: To investigate the effects of thyroxine treatment on development and growth of young Down syndrome children. DESIGN, SETTING, AND PARTICIPANTS: Single-center, randomized, double-blind, 24-month trial (enrollment June 1999 to August 2001) with nationwide recruitment, comparing thyroxine administration with placebo in 196 Down syndrome neonates. INTERVENTION: Neonates were randomly assigned to treatment for 2 yr with either thyroxine (n = 99; initial dose 8 microg/kg) or placebo (n = 97). Daily thyroxine doses were adjusted at regular intervals to maintain plasma TSH in its normal and free T(4) concentrations in its high-normal range. Placebo dose adjustments mirrored those of thyroxine. MAIN OUTCOME MEASURES: The primary outcome was mental and motor development at age 24 months, assessed with the Bayley Scales of Infant Development II. RESULTS: At age 24 months, the developmental testing results of 90 thyroxine-, and 91 placebo-treated children were available for analysis. The thyroxine-treated children had a 0.7-month smaller delay in motor developmental age (95% confidence interval, -1.4 to 0), corresponding to a difference of seven motor developmental index points. The mental developmental age delay was also 0.7 month smaller in the thyroxine group (95% confidence interval, -1.5 to 0.2), but lacked statistical significance. Thyroxine-treated children had greater gains in length (1.1 cm; 95% confidence interval, 0.2 to 2.0) and weight (378 g; 95% confidence interval 55 to 701). CONCLUSIONS: The data of our study provide evidence to support the hypothesis that thyroxine treatment may improve development and growth of young Down syndrome children. Thyroxine treatment should be considered in Down syndrome neonates to maximize their early development and growth. PMID- 15755849 TI - 15-Deoxy-{delta}12,14-prostaglandin j2 inhibits interleukin-1{beta}-induced nuclear factor-{kappa}b in human amnion and myometrial cells: mechanisms and implications. AB - Proinflammatory cytokines and prostaglandins play key roles in term and preterm human labor. The expression of the prostaglandin synthetic enzyme cyclooxygenase (COX)-2 and cytokines IL-1beta and IL-8 increases within the uterus at the time of labor, and each is regulated by the transcription factor nuclear factor-kappaB (NF-kappaB). In addition to its role in driving inflammation, COX-2 may also synthesize 15-deoxy-Delta (12, 14)-prostaglandin J(2) (15d-PGJ(2)), an antiinflammatory cyclopentenone prostaglandin (cyPG), which acts in some cells as an agonist of peroxisome proliferator-activated receptors (PPARs). We found that PPARalpha and -gamma proteins are expressed in both amnion epithelial and myometrial cells, but synthetic PPAR agonists could not inhibit NF-kappaB activity or COX-2 expression. 15d-PGJ(2) inhibited NF-kappaB activity and COX-2 expression in both cell types. This was unaffected by a PPAR antagonist and could be mimicked by the cyPG PGA(1) but not 9,10-dihydro-15d-PGJ(2) in which the cyclopentenone ring is disrupted. This shows that, in amnion and myometrium, inhibition of NF-kappaB activity and COX-2 expression by 15d-PGJ(2) is independent of PPARs and requires the cyclopentenone ring. We further show that 15d-PGJ(2) acts at multiple levels in the NF-kappaB pathway: blocking inhibitor of kappaBalpha degradation by repressing inhibitor of kappaB kinase activation and the 26S proteasome and also repressing NF-kappaB DNA binding and phosphorylation. Our data suggest that PPARs are unlikely to play a role in the regulation of either NF-kappaB or COX-2 in human amnion and myometrium. Targeting of NF-kappaB is a potential therapeutic strategy in preterm labor. PPAR agonists are unlikely to be effective in this context, but cyPGs may have potential. PMID- 15755848 TI - Congenital adrenal hyperplasia due to 11-hydroxylase deficiency: functional characterization of two novel point mutations and a three-base pair deletion in the CYP11B1 gene. AB - Congenital adrenal hyperplasia is a group of autosomal recessive disorders second most often caused by deficiency of steroid 11-hydroxylase (CYP11B1) due to mutations in the CYP11B1 gene. We studied the functional and structural consequences of two novel missense mutations (W116C, L299P) and an in-frame 3-bp deletion (DeltaF438) in the CYP11B gene, detected in three unrelated families. All patients are suffering from classical CYP11B1 deficiency. In vitro expression studies in COS-7 cells revealed a decreased CYP11B1 activity in the W116C mutant to 2.9 +/- 0.9% (sd) for the conversion of 11-deoxycortisol to cortisol. The L299P mutant reduced the enzymatic activity to 1.2 +/- 0.9%, whereas the DeltaF438 mutation resulted in no measurable residual CYP11B1 activity. Introduction of these mutations in a three-dimensional model structure of the CYP11B1 protein provides a possible explanation for the in vitro measured effects. We hypothesize that the W116C mutation influences the conformational change of the 11-hydroxylase protein necessary for substrate access and product release. The L299P mutation causes a change in the position of the I helix relative to the heme group, whereas the DeltaF438 mutation results in a steric disarrangement of the heme group relative to the enzyme. Studying the enzyme function in vitro helps to understand the phenotypical expression and disease severity of 11-hydroxylase deficiency, which is the basis for accurate genetic counseling, prenatal diagnosis, and treatment. Moreover, the combination of in vitro enzyme function and molecular modeling provides new insights in cytochrome P450 structural-functional relationships. PMID- 15755850 TI - Neutrophil count in small-for-gestational age children: contrasting effects of metformin and growth hormone therapy. AB - A minority of children born small for gestational age (SGA) maintain a slow weight gain and a short stature (SS). At the other end of the spectrum are SGA children who show rapid postnatal weight gain and catch-up growth; these subjects may develop hyperinsulinemia, exaggerated adrenarche with precocious pubarche (PP), and an associated proinflammatory state with raised IL-6 and reduced adiponectin levels. Metformin therapy in SGA-PP girls attenuates the hyperinsulinemia, the adrenal androgen excess, and the proinflammatory state. In contrast, GH therapy in SGA-SS children promotes height gain but may induce hyperinsulinemia. Both groups are associated with increased risk markers for future cardiovascular disease. Therefore, we studied markers of inflammation in both SGA subpopulations at baseline and after their respectively corrective therapies. SGA-PP girls (n = 33; mean age, 8 yr; body mass index, 18.5 kg/m(2)) were randomized to remain untreated or to receive metformin (425 mg/d) for 6 months. SGA-SS children (n = 29; mean age, 7 yr; body mass index, 14.7 kg/m(2)) were randomly assigned to remain untreated or to receive GH (60 mug/kg/d). In SGA PP girls, the mean neutrophil count (4.0 x 1000/mm(3)) was more than 2 sd above the mean reference level (2.8 x 1000/mm(3), P < 0.001); this remained stable over 6 months in untreated girls but dropped in metformin-treated girls by -1.1 x 1000/mm(3) (P = 0.002). In SGA-SS children, neutrophil counts were also higher at baseline (3.3 x 1000/mm(3), P < 0.01). This remained stable in untreated children but rose in GH-treated children by +1.1 x 1000/mm(3) (P = 0.004). GH-treated children also showed a rise in circulating IL-6 and dehydroepiandrosterone sulfate levels and a fall in adiponectin levels. In conclusion, neutrophil counts were elevated in SGA children. In SGA girls with PP, the present results corroborate the antiinflammatory benefits of metformin therapy. In contrast, high dose GH therapy in short SGA children may increase neutrophil counts and lead to a less favorable adipocytokine profile. Future studies with combined GH plus metformin treatment in short SGA children may clarify whether insulin resistance is a mechanism linking GH therapy to markers of inflammation. PMID- 15755851 TI - Iatrogenic Cushing's syndrome with osteoporosis and secondary adrenal failure in human immunodeficiency virus-infected patients receiving inhaled corticosteroids and ritonavir-boosted protease inhibitors: six cases. AB - Ritonavir, a protease inhibitor (PI), is a potent inhibitor of cytochrome P450 3A4. This pharmacological effect, even at low doses ( or vaccines). Factors associated with severe delay included having a mother who was unmarried or who did not have a college degree, living in a household with 2 or more children, being non-Hispanic black, having 2 or more vaccination providers, and using public vaccination provider(s). CONCLUSIONS: More than 1 in 3 children were undervaccinated for more than 6 months during their first 24 months of life and 1 in 4 children were delayed for at least 4 vaccines. Standard measures of vaccination coverage mask substantial shortfalls in ensuring that recommendations are followed regarding age at vaccination throughout the first 24 months of life. PMID- 15755942 TI - Role of computerized physician order entry systems in facilitating medication errors. AB - CONTEXT: Hospital computerized physician order entry (CPOE) systems are widely regarded as the technical solution to medication ordering errors, the largest identified source of preventable hospital medical error. Published studies report that CPOE reduces medication errors up to 81%. Few researchers, however, have focused on the existence or types of medication errors facilitated by CPOE. OBJECTIVE: To identify and quantify the role of CPOE in facilitating prescription error risks. DESIGN, SETTING, AND PARTICIPANTS: We performed a qualitative and quantitative study of house staff interaction with a CPOE system at a tertiary care teaching hospital (2002-2004). We surveyed house staff (N = 261; 88% of CPOE users); conducted 5 focus groups and 32 intensive one-on-one interviews with house staff, information technology leaders, pharmacy leaders, attending physicians, and nurses; shadowed house staff and nurses; and observed them using CPOE. Participants included house staff, nurses, and hospital leaders. MAIN OUTCOME MEASURE: Examples of medication errors caused or exacerbated by the CPOE system. RESULTS: We found that a widely used CPOE system facilitated 22 types of medication error risks. Examples include fragmented CPOE displays that prevent a coherent view of patients' medications, pharmacy inventory displays mistaken for dosage guidelines, ignored antibiotic renewal notices placed on paper charts rather than in the CPOE system, separation of functions that facilitate double dosing and incompatible orders, and inflexible ordering formats generating wrong orders. Three quarters of the house staff reported observing each of these error risks, indicating that they occur weekly or more often. Use of multiple qualitative and survey methods identified and quantified error risks not previously considered, offering many opportunities for error reduction. CONCLUSIONS: In this study, we found that a leading CPOE system often facilitated medication error risks, with many reported to occur frequently. As CPOE systems are implemented, clinicians and hospitals must attend to errors that these systems cause in addition to errors that they prevent. PMID- 15755944 TI - Chromosomal instability in amniocytes from fetuses of mothers who smoke. AB - CONTEXT: Tobacco increases the risk of systemic diseases, and it has adverse effects on pregnancy. However, only indirect data have been published on a possible genotoxic effect on pregnancy in humans. OBJECTIVES: To determine whether maternal smoking has a genotoxic effect on amniotic cells, expressed as an increased chromosomal instability, and to analyze whether any chromosomal regions are especially affected by exposure to tobacco. DESIGN, SETTING, AND PATIENTS: In this prospective study, amniocytes were obtained by routine amniocentesis for prenatal diagnosis from 25 controls and 25 women who smoke (> or =10 cigarettes/d for > or =10 years), who were asked to fill out a smoking questionnaire concerning their smoking habits. Chromosomal instability was analyzed in blinded fashion by 2 independent observers in routine chromosome spreads. Breakpoints implicated in chromosomal abnormalities were identified by G banding. MAIN OUTCOME MEASURES: Association between maternal smoking and increased chromosomal instability in amniotic fluid cells, expressed as chromosomal lesions (gaps and breaks) and structural chromosomal abnormalities. RESULTS: Comparison of cytogenetic data between smokers and nonsmokers (controls) showed important differences for the proportion of structural chromosomal abnormalities (smokers: 12.1% [96/793]; controls: 3.5% [26/752]; P = .002) and to a lesser degree for the proportion of metaphases with chromosomal instability (smokers: 10.5% [262/2492]; controls: 8.0% [210/2637]; P = .04), and for the proportion of chromosomal lesions (smokers: 15.7% [391/2492]; controls: 10.1% [267/2637]; P = .045). Statistical analysis of the 689 breakpoints detected showed that band 11q23, which is a band commonly implicated in hematopoietic malignancies, was the chromosomal region most affected by tobacco. CONCLUSIONS: Our findings show that smoking 10 or more cigarettes per day for at least 10 years and during pregnancy is associated with increased chromosomal instability in amniocytes. Band 11q23, known to be involved in leukemogenesis, seems especially sensitive to genotoxic compounds contained in tobacco. PMID- 15755945 TI - Effects of computerized clinical decision support systems on practitioner performance and patient outcomes: a systematic review. AB - CONTEXT: Developers of health care software have attributed improvements in patient care to these applications. As with any health care intervention, such claims require confirmation in clinical trials. OBJECTIVES: To review controlled trials assessing the effects of computerized clinical decision support systems (CDSSs) and to identify study characteristics predicting benefit. DATA SOURCES: We updated our earlier reviews by searching the MEDLINE, EMBASE, Cochrane Library, Inspec, and ISI databases and consulting reference lists through September 2004. Authors of 64 primary studies confirmed data or provided additional information. STUDY SELECTION: We included randomized and nonrandomized controlled trials that evaluated the effect of a CDSS compared with care provided without a CDSS on practitioner performance or patient outcomes. DATA EXTRACTION: Teams of 2 reviewers independently abstracted data on methods, setting, CDSS and patient characteristics, and outcomes. DATA SYNTHESIS: One hundred studies met our inclusion criteria. The number and methodologic quality of studies improved over time. The CDSS improved practitioner performance in 62 (64%) of the 97 studies assessing this outcome, including 4 (40%) of 10 diagnostic systems, 16 (76%) of 21 reminder systems, 23 (62%) of 37 disease management systems, and 19 (66%) of 29 drug-dosing or prescribing systems. Fifty-two trials assessed 1 or more patient outcomes, of which 7 trials (13%) reported improvements. Improved practitioner performance was associated with CDSSs that automatically prompted users compared with requiring users to activate the system (success in 73% of trials vs 47%; P = .02) and studies in which the authors also developed the CDSS software compared with studies in which the authors were not the developers (74% success vs 28%; respectively, P = .001). CONCLUSIONS: Many CDSSs improve practitioner performance. To date, the effects on patient outcomes remain understudied and, when studied, inconsistent. PMID- 15755946 TI - The unintended consequences of publicly reporting quality information. AB - Health care report cards publicly report information about physician, hospital, and health plan quality in an attempt to improve that quality. Reporting quality information publicly is presumed to motivate quality improvement through 2 main mechanisms. First, public quality information allows patients, referring physicians, and health care purchasers to preferentially select high-quality physicians. Second, public report cards may motivate physicians to compete on quality and, by providing feedback and by identifying areas for quality improvement initiatives, help physicians to do so. Despite these plausible mechanisms of quality improvement, the value of publicly reporting quality information is largely undemonstrated and public reporting may have unintended and negative consequences on health care. These unintended consequences include causing physicians to avoid sick patients in an attempt to improve their quality ranking, encouraging physicians to achieve "target rates" for health care interventions even when it may be inappropriate among some patients, and discounting patient preferences and clinical judgment. Public reporting of quality information promotes a spirit of openness that may be valuable for enhancing trust of the health professions, but its ability to improve health remains undemonstrated, and public reporting may inadvertently reduce, rather than improve, quality. Given these limitations, it may be necessary to reassess the role of public quality reporting in quality improvement. PMID- 15755948 TI - Screening for psychological illness in military personnel. PMID- 15755949 TI - Computer technology and clinical work: still waiting for Godot. PMID- 15755950 TI - Smoking while pregnant: transplacental mutagenesis of the fetus by tobacco smoke. PMID- 15755947 TI - Screening for breast cancer. AB - CONTEXT: Breast cancer screening in community practices may be different from that in randomized controlled trials. New screening modalities are becoming available. OBJECTIVES: To review breast cancer screening, especially in the community and to examine evidence about new screening modalities. DATA SOURCES AND STUDY SELECTION: English-language articles of randomized controlled trials assessing effectiveness of breast cancer screening were reviewed, as well as meta analyses, systematic reviews, studies of breast cancer screening in the community, and guidelines. Also, studies of newer screening modalities were assessed. DATA SYNTHESIS: All major US medical organizations recommend screening mammography for women aged 40 years and older. Screening mammography reduces breast cancer mortality by about 20% to 35% in women aged 50 to 69 years and slightly less in women aged 40 to 49 years at 14 years of follow-up. Approximately 95% of women with abnormalities on screening mammograms do not have breast cancer with variability based on such factors as age of the woman and assessment category assigned by the radiologist. Studies comparing full-field digital mammography to screen film have not shown statistically significant differences in cancer detection while the impact on recall rates (percentage of screening mammograms considered to have positive results) was unclear. One study suggested that computer-aided detection increases cancer detection rates and recall rates while a second larger study did not find any significant differences. Screening clinical breast examination detects some cancers missed by mammography, but the sensitivity reported in the community is lower (28% to 36%) than in randomized trials (about 54%). Breast self-examination has not been shown to be effective in reducing breast cancer mortality, but it does increase the number of breast biopsies performed because of false-positives. Magnetic resonance imaging and ultrasound are being studied for screening women at high risk for breast cancer but are not recommended for screening the general population. Sensitivity of magnetic resonance imaging in high-risk women has been found to be much higher than that of mammography but specificity is generally lower. Effect of the magnetic resonance imaging on breast cancer mortality is not known. A balanced discussion of possible benefits and harms of screening should be undertaken with each woman. CONCLUSIONS: In the community, mammography remains the main screening tool while the effectiveness of clinical breast examination and self-examination are less. New screening modalities are unlikely to replace mammography in the near future for screening the general population. PMID- 15755951 TI - JAMA patient page. Smoking and pregnancy. PMID- 15755953 TI - Diversifying carotenoid biosynthetic pathways by directed evolution. AB - Microorganisms and plants synthesize a diverse array of natural products, many of which have proven indispensable to human health and well-being. Although many thousands of these have been characterized, the space of possible natural products--those that could be made biosynthetically--remains largely unexplored. For decades, this space has largely been the domain of chemists, who have synthesized scores of natural product analogs and have found many with improved or novel functions. New natural products have also been made in recombinant organisms, via engineered biosynthetic pathways. Recently, methods inspired by natural evolution have begun to be applied to the search for new natural products. These methods force pathways to evolve in convenient laboratory organisms, where the products of new pathways can be identified and characterized in high-throughput screening programs. Carotenoid biosynthetic pathways have served as a convenient experimental system with which to demonstrate these ideas. Researchers have mixed, matched, and mutated carotenoid biosynthetic enzymes and screened libraries of these "evolved" pathways for the emergence of new carotenoid products. This has led to dozens of new pathway products not previously known to be made by the assembled enzymes. These new products include whole families of carotenoids built from backbones not found in nature. This review details the strategies and specific methods that have been employed to generate new carotenoid biosynthetic pathways in the laboratory. The potential application of laboratory evolution to other biosynthetic pathways is also discussed. PMID- 15755952 TI - The acetate switch. AB - To succeed, many cells must alternate between life-styles that permit rapid growth in the presence of abundant nutrients and ones that enhance survival in the absence of those nutrients. One such change in life-style, the "acetate switch," occurs as cells deplete their environment of acetate-producing carbon sources and begin to rely on their ability to scavenge for acetate. This review explains why, when, and how cells excrete or dissimilate acetate. The central components of the "switch" (phosphotransacetylase [PTA], acetate kinase [ACK], and AMP-forming acetyl coenzyme A synthetase [AMP-ACS]) and the behavior of cells that lack these components are introduced. Acetyl phosphate (acetyl approximately P), the high-energy intermediate of acetate dissimilation, is discussed, and conditions that influence its intracellular concentration are described. Evidence is provided that acetyl approximately P influences cellular processes from organelle biogenesis to cell cycle regulation and from biofilm development to pathogenesis. The merits of each mechanism proposed to explain the interaction of acetyl approximately P with two-component signal transduction pathways are addressed. A short list of enzymes that generate acetyl approximately P by PTA ACKA-independent mechanisms is introduced and discussed briefly. Attention is then directed to the mechanisms used by cells to "flip the switch," the induction and activation of the acetate-scavenging AMP-ACS. First, evidence is presented that nucleoid proteins orchestrate a progression of distinct nucleoprotein complexes to ensure proper transcription of its gene. Next, the way in which cells regulate AMP-ACS activity through reversible acetylation is described. Finally, the "acetate switch" as it exists in selected eubacteria, archaea, and eukaryotes, including humans, is described. PMID- 15755954 TI - Signaling by target of rapamycin proteins in cell growth control. AB - Target of rapamycin (TOR) proteins are members of the phosphatidylinositol kinase related kinase (PIKK) family and are highly conserved from yeast to mammals. TOR proteins integrate signals from growth factors, nutrients, stress, and cellular energy levels to control cell growth. The ribosomal S6 kinase 1 (S6K) and eukaryotic initiation factor 4E binding protein 1(4EBP1) are two cellular targets of TOR kinase activity and are known to mediate TOR function in translational control in mammalian cells. However, the precise molecular mechanism of TOR regulation is not completely understood. One of the recent breakthrough studies in TOR signaling resulted in the identification of the tuberous sclerosis complex gene products, TSC1 and TSC2, as negative regulators for TOR signaling. Furthermore, the discovery that the small GTPase Rheb is a direct downstream target of TSC1-TSC2 and a positive regulator of the TOR function has significantly advanced our understanding of the molecular mechanism of TOR activation. Here we review the current understanding of the regulation of TOR signaling and discuss its function as a signaling nexus to control cell growth during normal development and tumorigenesis. PMID- 15755959 TI - Prognostic and predictive factors for breast cancer: translating technology to oncology. PMID- 15755955 TI - Initiation of protein synthesis in bacteria. AB - Valuable information on translation initiation is available from biochemical data and recently solved structures. We present a detailed description of current knowledge about the structure, function, and interactions of the individual components involved in bacterial translation initiation. The first section describes the ribosomal features relevant to the initiation process. Subsequent sections describe the structure, function, and interactions of the mRNA, the initiator tRNA, and the initiation factors IF1, IF2, and IF3. Finally, we provide an overview of mechanisms of regulation of the translation initiation event. Translation occurs on ribonucleoprotein complexes called ribosomes. The ribosome is composed of a large subunit and a small subunit that hold the activities of peptidyltransfer and decode the triplet code of the mRNA, respectively. Translation initiation is promoted by IF1, IF2, and IF3, which mediate base pairing of the initiator tRNA anticodon to the mRNA initiation codon located in the ribosomal P-site. The mechanism of translation initiation differs for canonical and leaderless mRNAs, since the latter is dependent on the relative level of the initiation factors. Regulation of translation occurs primarily in the initiation phase. Secondary structures at the mRNA ribosomal binding site (RBS) inhibit translation initiation. The accessibility of the RBS is regulated by temperature and binding of small metabolites, proteins, or antisense RNAs. The future challenge is to obtain atomic-resolution structures of complete initiation complexes in order to understand the mechanism of translation initiation in molecular detail. PMID- 15755960 TI - Chemoprevention in the 21st century: is a balance best or should women have no estrogen at all? PMID- 15755956 TI - Cellulase, clostridia, and ethanol. AB - Biomass conversion to ethanol as a liquid fuel by the thermophilic and anaerobic clostridia offers a potential partial solution to the problem of the world's dependence on petroleum for energy. Coculture of a cellulolytic strain and a saccharolytic strain of Clostridium on agricultural resources, as well as on urban and industrial cellulosic wastes, is a promising approach to an alternate energy source from an economic viewpoint. This review discusses the need for such a process, the cellulases of clostridia, their presence in extracellular complexes or organelles (the cellulosomes), the binding of the cellulosomes to cellulose and to the cell surface, cellulase genetics, regulation of their synthesis, cocultures, ethanol tolerance, and metabolic pathway engineering for maximizing ethanol yield. PMID- 15755961 TI - Integration of imaging in the management of breast cancer. PMID- 15755962 TI - Continuing evolution in breast cancer surgical management. PMID- 15755963 TI - Radiation therapy for invasive breast cancer: not just for local control. PMID- 15755964 TI - Optimizing endocrine therapy for breast cancer. PMID- 15755965 TI - Moving away from the "one shoe fits all" strategy: the key to future progress in chemotherapy. PMID- 15755966 TI - What is targeted therapy? PMID- 15755967 TI - Estrogen-receptor biology: continuing progress and therapeutic implications. PMID- 15755968 TI - Circulating and disseminated tumor cells. PMID- 15755957 TI - Detection of and response to signals involved in host-microbe interactions by plant-associated bacteria. AB - Diverse interactions between hosts and microbes are initiated by the detection of host-released chemical signals. Detection of these signals leads to altered patterns of gene expression that culminate in specific and adaptive changes in bacterial physiology that are required for these associations. This concept was first demonstrated for the members of the family Rhizobiaceae and was later found to apply to many other plant-associated bacteria as well as to microbes that colonize human and animal hosts. The family Rhizobiaceae includes various genera of rhizobia as well as species of Agrobacterium. Rhizobia are symbionts of legumes, which fix nitrogen within root nodules, while Agrobacterium tumefaciens is a pathogen that causes crown gall tumors on a wide variety of plants. The plant-released signals that are recognized by these bacteria are low-molecular weight, diffusible molecules and are detected by the bacteria through specific receptor proteins. Similar phenomena are observed with other plant pathogens, including Pseudomonas syringae, Ralstonia solanacearum, and Erwinia spp., although here the signals and signal receptors are not as well defined. In some cases, nutritional conditions such as iron limitation or the lack of nitrogen sources seem to provide a significant cue. While much has been learned about the process of host detection over the past 20 years, our knowledge is far from being complete. The complex nature of the plant-microbe interactions makes it extremely challenging to gain a comprehensive picture of host detection in natural environments, and thus many signals and signal recognition systems remain to be described. PMID- 15755969 TI - Physician/patient decision aids for adjuvant therapy. PMID- 15755970 TI - Gene expression profiling of breast cancer: a new tumor marker. PMID- 15755971 TI - Aromatase inhibitors for breast cancer prevention. PMID- 15755972 TI - Selective estrogen-receptor modulators for primary prevention of breast cancer. PMID- 15755973 TI - Prevention and management of hereditary breast cancer. PMID- 15755974 TI - FDG-PET and beyond: molecular breast cancer imaging. PMID- 15755975 TI - Future directions in breast imaging. PMID- 15755976 TI - Magnetic resonance imaging of the breast: opportunities to improve breast cancer management. PMID- 15755977 TI - Advances in breast conservation therapy. PMID- 15755978 TI - Lymphatic mapping and sentinel lymph node biopsy for breast cancer: developments and resolving controversies. PMID- 15755979 TI - The use of radiotherapy after mastectomy: a review of the literature. PMID- 15755980 TI - Use of conventional radiation therapy as part of breast-conserving treatment. PMID- 15755981 TI - Accelerated partial breast irradiation as a part of breast conservation therapy. PMID- 15755982 TI - Adjuvant endocrine therapy for premenopausal women with early breast cancer. PMID- 15755983 TI - Advances in adjuvant hormonal therapy for postmenopausal women. PMID- 15755984 TI - Chemotherapy: what progress in the last 5 years? PMID- 15755985 TI - Targeted therapy: wave of the future. PMID- 15755986 TI - Angiogenesis of breast cancer. PMID- 15755987 TI - A randomized, double-blinded, placebo-controlled phase II trial of recombinant human leukemia inhibitory factor (rhuLIF, emfilermin, AM424) to prevent chemotherapy-induced peripheral neuropathy. PMID- 15755988 TI - AKR1B10: a new diagnostic marker of non-small cell lung carcinoma in smokers. PMID- 15755989 TI - Prevention of chemotherapy-induced neuropathy: leukemia inhibitory factor. PMID- 15755990 TI - Identification of a 40S ribosomal protein S4-derived H-Y epitope able to elicit a lymphoblast-specific cytotoxic T lymphocyte response. AB - PURPOSE: The superior graft-versus-leukemia (GVL) effect of the female-to-male stem cell transplantation is partially independent from the concomitant graft versus-host reactivity. However, the antigenic basis of this selective GVL response remains enigmatic, because no H-Y antigens with hematopoietic-restricted expression were identified. In this study, we report a novel H-Y epitope that is preferentially recognized on activated proliferating lymphocytes. EXPERIMENTAL DESIGN: We generated a CTL clone YKIII.8 that showed reactivity toward male B*5201+ CD40-activated B cells, EBV-lymphoblastoid cell lines, and phytohemagglutinin-activated T-cell blasts but little or no reactivity toward fibroblasts, CD14+ cells, or unstimulated B and T cells. The antigen recognized by YKIII.8 was identified by screening of a cDNA expression library, and its pattern of expression was investigated. RESULTS: cDNA of the male isoform of 40S ribosomal protein S4 was found to encode the antigenic peptide TIRYPDPVI, which was recognized by YKIII.8. Western blot analysis showed that rapidly proliferating cells overexpress the RPS4 protein in comparison with nonrecognized cell subsets. Retroviral transfer of YKIII.8 T-cell receptor resulted in preservation of the lymphoblast-specific reactivity pattern. CONCLUSION: Our findings suggest that CTL specific to certain epitopes of ubiquitously expressed H-Y antigens may specifically target lymphoblasts, contributing to the selective GVL effect of female-to-male stem cell transplantation. PMID- 15755991 TI - Heregulin-induced activation of HER2 and HER3 increases androgen receptor transactivation and CWR-R1 human recurrent prostate cancer cell growth. AB - PURPOSE: The androgen receptor (AR) is a ligand-dependent transcription factor that mediates gene expression and growth of normal and malignant prostate cells. In prostate tumors that recur after androgen withdrawal, the AR is highly expressed and transcriptionally active in the absence of testicular androgens. In these "androgen-independent" tumors, alternative means of AR activation have been invoked, including regulation by growth factors and their receptors in prostate cancer recurrence. EXPERIMENTAL DESIGN AND RESULTS: In this report, we show that HER receptor tyrosine kinases 1 through 4 are expressed in the CWR-R1 recurrent prostate cancer cell line; their stimulation by epidermal growth factor (EGF) and heregulin activates downstream signaling, including mitogen-activated protein kinase and phosphatidylinositol-3 kinase and Akt pathways. We show that heregulin activates HER2 and HER3 and increases androgen-dependent AR transactivation of reporter genes in CWR-R1 cells. Tyrosine phosphorylation of HER2 and HER3, AR transactivation, and cell proliferation induced by heregulin were more potently inhibited by the EGFR/HER2 dual tyrosine kinase inhibitor GW572016 (lapatinib) than the EGFR-specific inhibitor ZD1839 (gefitinib). Basal proliferation in the absence of growth factors was also inhibited by GW572016 to a greater extent than ZD1839, suggesting that low level HER2/HER3 activation perhaps by an autocrine pathway contributes to the proliferation signal. CONCLUSIONS: These data indicate that heregulin signaling through HER2 and HER3 increases AR transactivation and alters growth in a recurrent prostate cancer cell line. Therefore, inhibition of low-level HER2 signaling may be a potential novel therapeutic strategy in prostate cancer. PMID- 15755992 TI - An orthotopic model of anaplastic thyroid carcinoma in athymic nude mice. AB - PURPOSE: To develop an orthotopic model of anaplastic thyroid carcinoma (ATC) in athymic nude mice. EXPERIMENTAL DESIGN: Various thyroid carcinoma cell lines were injected into the thyroid gland of athymic nude mice to determine whether such injection was technically feasible. ATC cells were then injected into the thyroid gland or the subcutis of nude mice at various concentrations, and the mice were then followed for tumor development. The tumors were examined histopathologically for local invasion or regional or distant metastasis. RESULTS: Injection of tumor cells into the thyroid glands of nude mice was technically feasible and resulted in the formation of thyroid tumors. The ATC cell line DRO showed significantly higher tumorigenicity in the thyroid gland than in the subcutis. In contrast, oral squamous cell carcinoma cell line TU167 shows no significantly higher tumorigenicity in the thyroid gland than in the subcutis. ATC tumors established in the thyroid gland also produced symptomatic compression of the esophagus and the trachea. Local invasion of the larynx and trachea was as well as high rates of pulmonary metastasis were also observed. Immunohistochemical staining showed higher microvessel density as well as higher expression of vascular endothelial growth factor and interleukin-8 in the orthotopic thyroid tumors than in ectopic tumors. CONCLUSION: An orthotopic model of ATC in athymic nude mice was developed that closely recapitulates the clinical findings of human ATC. This model should facilitate the understanding of the pathogenesis of ATC and aid in the development of novel therapies against ATC. PMID- 15755993 TI - 2-methoxyestradiol is an estrogen receptor agonist that supports tumor growth in murine xenograft models of breast cancer. AB - PURPOSE: 2-Methoxyestradiol (2MEO) is being developed as a novel antitumor agent based on its antiangiogenic activity, tumor cell cytotoxicity, and apparent lack of toxicity. However, pharmacologic concentrations of 2MEO bind to estrogen receptors (ER). We have therefore examined the ER activity of 2MEO. EXPERIMENTAL DESIGN: Estrogenic actions of 2MEO were evaluated by changes in gene expression of the ER-positive (MCF7) breast tumor cell line and, in vivo, estrogenicity was assessed in breast tumor xenograft models and by measuring endocrine responses in uterus and liver. RESULTS: In the ER-positive breast tumor cell line (MCF7), microarray experiments revealed that 269 of 279 changes in gene expression common to 2MEO and estradiol were prevented by the ER antagonist, ICI 182,780. Changes in the expression of selected genes and their sensitivity to inhibition by ICI 182,780 were confirmed by quantitative reverse transcription-PCR measurement. Activation of ER in MCF7 cells by 2MEO was further confirmed by stimulation of an estrogen response element-dependent reporter gene that was blocked by ICI 182,780 (1 micromol/L). Doses of 2MEO (15-150 mg/kg) that had no antitumor efficacy in either nu/nu BALB/c or severe combined immunodeficient mice bearing ER-negative MDA-MB-435 tumors had uterotropic and hepatic estrogen-like actions. In female nu/nu BALB/c mice inoculated with the estrogen-dependent MCF7 tumor cells, 2MEO (50 mg/kg/d) supported tumor growth. CONCLUSIONS: Tumor growth enhancement by 2MEO at doses generating serum levels (100-500 nmol/L) that have estrogenic activity suggests that a conservative approach to the further clinical evaluation of this agent should be adopted and that its evaluation in breast cancer is inappropriate. PMID- 15755994 TI - Influence of CD80, interleukin-2, and interleukin-7 expression in human renal cell carcinoma on the expansion, function, and survival of tumor-specific CTLs. AB - PURPOSE: A renal cell carcinoma (RCC) line, RCC-26, has been identified as a suitable candidate for development of an allogeneic tumor cell vaccine based on its expression of a variety of tumor-associated antigens (TAA). To improve immunogenicity, RCC-26 cells were genetically engineered to express CD80 alone or in combination with interleukin (IL)-2 or IL-7. The effect of these modifications on proliferation, function, and survival of autologous and allogeneic tumor specific CTLs was assessed. EXPERIMENTAL DESIGN: RCC-26 sublines expressing different transgenes were tested for their capacity to reactivate cytokine secretion and cytotoxicity in autologous tumor-infiltrating lymphocytes, to improve proliferation and survival of tumor-associated T cells present in autologous peripheral blood, and to induce tumor-associated responses in naive allogeneic lymphocytes. The expression of several common TAA was quantitated in the RCC-26 sublines using reverse transcription-PCR to identify surrogate markers for immune monitoring in clinical trials. RESULTS: Gene-modified RCC-26 cells showed enhanced immunogenicity. CD80 expression was necessary to induce RCC associated CTL in blood of healthy allogeneic donors. It also improved proliferation of autologous effector-memory T cells. Further enhancement was achieved with IL-2 through induction of the antiapoptosis protein Bcl-x(L). The candidate vaccine lines overexpressed several common TAA that are suitable markers for immune monitoring. CONCLUSIONS: RCC-26 cells coexpressing CD80 and cytokine transgenes display improved immunogenic characteristics, supporting their use as allogeneic tumor cell vaccines for HLA-A2-matched patients with metastatic RCC. PMID- 15755995 TI - Effect of chemokine receptors CXCR4 and CCR7 on the metastatic behavior of human colorectal cancer. AB - PURPOSE: The expression of chemokine receptors CXCR4 and CCR7 has been associated with tumor dissemination and poor prognosis in a limited number of tumor entities. However, no data are currently available on the impact of chemokine receptor expression on disease progression and prognosis in human colorectal cancer. EXPERIMENTAL DESIGN: The expression of CXCR4 and CCR7 was evaluated in 96 patients with histologically confirmed colorectal cancers and in four colorectal cancer cell lines by immunohistochemical staining. Furthermore, cell migration assays were done with SW480, SW620, and LS174T cancer cells to confirm the effect of the CXCR4 ligand stromal cell-derived factor 1alpha on migration. RESULTS: Human colorectal cancer specimens and cell lines displayed a CXCR4 and CCR7 expression with variable intensities. Interestingly, strong expression of CXCR4, but not of CCR7, was significantly associated with higher Union International Contre Cancer stages 3/4 (P = 0.0017), lymph node metastasis (P = 0.00375), and distant metastasis (P = 0.00003) and further correlated with a reduced 3-year survival rate (P = 0.1). Strong CXCR4 and CCR7 expression positively correlated with the location of the primary tumor in the rectum (P < 0.01). Furthermore, activation of CXCR4-expressing cancer cells by stromal cell-derived factor 1alpha resulted in a significant increase of cell migration (P < 0.014). CONCLUSION: Strong expression of CXCR4 by colorectal cancer cells is significantly associated with lymphatic and distant dissemination in patients with colorectal cancer as well as with cancer cell migration in vitro. PMID- 15755997 TI - Elevated serum cytokines correlated with altered behavior, serum cortisol rhythm, and dampened 24-hour rest-activity patterns in patients with metastatic colorectal cancer. AB - PURPOSE: Incapacitating symptom burden in cancer patients contributes to poor quality of life (QOL) and can influence treatment outcomes because of poor tolerance to therapy. In this study, the role of circulating cytokines in the production symptoms in cancer patients is evaluated. EXPERIMENTAL DESIGN: Eighty patients with metastatic colorectal cancer with either normal (group I, n = 40) or dampened (group II, n = 40) 24-hour rest/activity patterns measured by actigraphy were identified. Actigraphy patterns were correlated with QOL indices, serum cortisol obtained at 8:00 a.m. and 4:00 p.m. and with serum levels of transforming growth factor-alpha, tumor necrosis factor-alpha, and interleukin 6 (IL-6) obtained at 8:00 a.m. and analyzed in duplicate by ELISA. Cytokine levels and survival were also correlated. RESULTS: Group II patients had significantly higher pre treatment levels of all three cytokines, displayed significantly poorer emotional and social functioning, had higher fatigue, more appetite loss, and poorer performance status compared with group I patients. Transforming growth factor-alpha (TGF-alpha) and IL-6 were significantly increased in the patients with WHO performance status >1 and in those with appetite loss. Fatigue was significantly associated with elevated TGF-alpha only. IL-6 was increased in those patients with extensive liver involvement and multiple organ replacement, and it was significantly correlated with dampened cortisol rhythm. In a multivariate analysis, IL-6 was correlated with poor treatment outcome. CONCLUSIONS: Significant correlations were found between serum levels of TGF alpha and IL-6, circadian patterns in wrist activity and serum cortisol and tumor related symptoms in patients with metastatic colorectal cancer. These data support the hypothesis that some cancer patient's symptoms of fatigue, poor QOL, and treatment outcome are related to tumor or host generated cytokines and could reflect cytokine effects on the circadian timing system. This interplay between cytokine signaling pathways, the hypothalamic-pituitary-adrenal axis, the autonomic nervous system, and efferent pathways of the suprachiasmatic nucleus that control circadian physiology, opens the way to new rational interventions for symptom management in cancer patients. PMID- 15755996 TI - Up-regulation of the peripheral benzodiazepine receptor during human colorectal carcinogenesis and tumor spread. AB - The peripheral benzodiazepine receptor (PBR) is overexpressed in a variety of cancers. In Unio Internationale Contra Cancrum (UICC) III colorectal cancers, a high level of PBR overexpression correlates with poor prognosis. However, little is known about the role of PBR in the development and progression of colorectal cancer. This study addresses the up-regulation of PBR during colorectal carcinogenesis and tumor spread. One hundred sixteen consecutive patients undergoing surgery for colorectal cancer with either regional (59 patients) or distant metastases (57 patients) were followed-up for 5 years or until death. Twenty-four of the 59 patients with initial UICC stage III cancers later developed distant metastases. PBR overexpression in tumor specimens was determined by immunohistochemistry. UICC stage III patients with colorectal primaries highly overexpressing PBR developed metastases significantly more often than patients with low PBR overexpression in their primary carcinoma. In 54 of the 116 patients adenomas and/or metastases and/or recurrences were available to be studied for PBR up-regulation during colorectal carcinogenesis and tumor spread. PBR was found to be overexpressed in 86% of early and late adenomas. Furthermore, 85% of primaries and of 86% of metastases displayed PBR overexpression. PBR overexpression was also detected at the mRNA level as revealed by real-time PCR. The extent of PBR protein overexpression was equivalent in colorectal adenomas and carcinomas but slightly increased in metastases. These data suggest a functional role of PBR during colorectal carcinogenesis and tumor spread. Thus, PBR qualifies as a target for innovative diagnostic and therapeutic approaches. PMID- 15755998 TI - Allelotypes and fluorescence in situ hybridization profiles of poorly differentiated endocrine carcinomas of different sites. AB - PURPOSE: The aim of this work was to investigate the genotypic profiles of 36 poorly differentiated endocrine carcinoma (PDEC) of different sites to verify if their very similar phenotype may reflect similar pattern of genetic anomalies and if useful diagnostic or prognostic markers may be pointed out. EXPERIMENTAL DESIGN: All tumors were microallelotyped at 57 microsatellite on 11 autosomes and the allelotypes of a selected panel of tumors were validated by interphasic fluorescence in situ hybridization with centromeric probes for chromosomes 1, 3, 6, 11, 17, and 18 and a probe specific for p53. RESULTS: Regardless of the primary sites, PDECs exhibit very complex allelotypes (86%) and TP53 allelic imbalance (89%). Among these cases, fluorescence in situ hybridization analysis confirmed the presence of multiple aneusomies and a chromosome instability phenotype. Very low percentage of allelic imbalance (AI) and few aneuploidies were detected in only five PDECs for which an overall longer survival was observed. We found recurrent AI on 3p, 5, and 11q13 in lung PDECs, on 5q21, 8p, and 18q21 in colorectal PDECs and on 7 and 11q22 in gastric PDECs. Significantly better outcome was observed in patients with PDEC exhibiting 8q AIs and absence of AI at chromosome regions 6q25 and 6p. CONCLUSIONS: The concurrence of p53 inactivation and aneuploidies or chromosome instability are the main features of PDECs. However, the specific allelotypes observed in relation to primary site support the hypothesis that PDECs and exocrine carcinomas of all sites may share early pathogenetic mechanisms. Molecular markers of potential diagnostic and prognostic values for PDECs of different sites have been identified. PMID- 15755999 TI - Overexpression of the aldo-keto reductase family protein AKR1B10 is highly correlated with smokers' non-small cell lung carcinomas. AB - PURPOSE: Squamous cell carcinoma (SCC) and adenocarcinoma of the lung are currently subject to similar treatment regimens despite distinct differences in histology and epidemiology. The aim of this study is to identify a molecular target with diagnostic and therapeutic values for SCC. EXPERIMENTAL DESIGN: Genes specifically up-regulated in SCC were explored through microarray analysis of 5 SCCs, 5 adenocarcinomas, 10 small cell lung carcinomas, 27 normal tissues, and 40 cancer cell lines. Clinical usefulness of these genes was subsequently examined mainly by immunohistochemical analysis. RESULTS: Seven genes, including aldo-keto reductase family 1, member B10 (AKR1B10), were identified as SCC-specific genes. AKR1B10 was further examined by immunohistochemical analysis of 101 non-small cell lung carcinomas (NSCLC) and its overexpression was observed in 27 of 32 (84.4%) SCCs and 19 of 65 (29.2%) adenocarcinomas. Multiple regression analysis showed that smoking was an independent variable responsible for AKR1B10 overexpression in NSCLCs (P < 0.01) and adenocarcinomas (P < 0.01). AKR1B10 staining was occasionally observed even in squamous metaplasia, a precancerous lesion of SCC. CONCLUSION: AKR1B10 was overexpressed in most cases with SCC, which is closely associated with smoking, and many adenocarcinoma cases of smokers. These results suggest that AKR1B10 is a potential diagnostic marker specific to smokers' NSCLCs and might be involved in tobacco-related carcinogenesis. PMID- 15756000 TI - Prognostic validation of the international classification of immunoglobulin M gammopathies: a survival advantage for patients with immunoglobulin M monoclonal gammopathy of undetermined significance? AB - PURPOSE: To verify the reliability of the new criteria for the diagnosis of IgM gammopathies recently proposed by an international panel of experts (Athens, 2002). EXPERIMENTAL DESIGN: A retrospective series of 698 patients with IgM gammopathy was reviewed paying attention to symptoms, serum IgM concentration, bone marrow infiltration, blood cell count and clinical course. Four clinical entities can be identified: IgM monoclonal gammopathy of undetermined significance (IgM-MGUS), asymptomatic and symptomatic Wandenstrom's macroglobulinemia (A-WM and S-WM, respectively), and IgM-related disorders, although this last was excluded from the study because of the scarcity of patients due to probable selection biases. The observed mortality was studied related to that expected in the general population of comparable age and sex and over an equivalent period of follow-up (standardized mortality ratio, SMR). RESULTS: IgM-MGUS, A-WM, and S-WM shared many clinical aspects but, with respect to the general population, patients with IgM-MGUS had a slight but definite survival advantage, those with A-WM had a mortality rate equivalent to that of the general population, whereas the SMR of patients with S-WM was 5.4. Within A WM and S-WM the SMR values did not vary significantly in relation to marrow lymphocyte counts or serum IgM concentrations. CONCLUSIONS: Our findings represent a prognostic validation of the applied diagnostic criteria for three of the four identifiable clinical entities and highlight the importance of symptoms over serum IgM concentration and marrow infiltration. PMID- 15756002 TI - Neutral endopeptidase 24.11/CD10 suppresses progressive potential in ovarian carcinoma in vitro and in vivo. AB - Recently, numerous studies have shown that endothelin-1 (ET-1) is expressed in ovarian carcinoma and that ET-1 selectively acts as an autocrine or paracrine growth factor through the endothelin A receptor (ET(A)R), and is involved in cell proliferation, invasiveness, neovascularization, and prevention of apoptosis. Neutral endopeptidase 24.11 (NEP) is a cell surface aminopeptidase with a ubiquitous expression and is capable of degrading a number of bioactive peptides including ET-1. Our previous report showed that stromal NEP expression in ovarian carcinoma was down-regulated as the histologic grade advanced. Here, we confirmed that NEP was expressed in tumor cells as well as stromal tissues in ovarian carcinoma, and investigated the functions of NEP in this carcinoma. We showed that there was a significant decrease in cell proliferation and invasiveness with a reduction in the concentration of ET-1 in the conditioned medium on the NEP overexpression of NEP in ovarian carcinoma cells. In addition, the overexpression of NEP enhanced susceptibility to paclitaxel, resulting in an increased occurrence of apoptotic morphologic change. Furthermore, tumorigenesis was reduced in vivo with the overexpression of NEP, down-regulation of both matrix metalloproteinase-2, and vascular endothelial growth factor expression. This evidence suggests that NEP functionally suppresses the progression of ovarian carcinoma and further study of this enzyme may reveal an effective way to target ET-1 for the treatment of this carcinoma. PMID- 15756001 TI - Fractional genomic alteration detected by array-based comparative genomic hybridization independently predicts survival after hepatic resection for metastatic colorectal cancer. AB - PURPOSE: Although liver resection is the primary curative therapy for patients with colorectal hepatic metastases, most patients have a recurrence. Identification of molecular markers that predict patients at highest risk for recurrence may help to target further therapy. EXPERIMENTAL DESIGN: Array-based comparative genomic hybridization was used to investigate the association of DNA copy number alterations with outcome in patients with colorectal liver metastasis resected with curative intent. DNA from 50 liver metastases was labeled and hybridized onto an array consisting of 2,463 bacterial artificial chromosome clones covering the entire genome. The total fraction of genome altered (FGA) in the metastases and the patient's clinical risk score (CRS) were calculated to identify independent prognostic factors for survival. RESULTS: An average of 30 +/- 14% of the genome was altered in the liver metastases (14% gained and 16% lost). As expected, a lower CRS was an independent predictor of overall survival (P = 0.03). In addition, a high FGA also was an independent predictor of survival (P = 0.01). The median survival time in patients with a low CRS (score 0-2) and a high (> or =20%) FGA was 38 months compared with 18 months in patients with a low CRS and a low FGA. Supervised analyses, using Prediction Analysis of Microarrays and Significance Analysis of Microarrays, identified a set of clones, predominantly located on chromosomes 7 and 20, which best predicted survival. CONCLUSIONS: Both FGA and CRS are independent predictors of survival in patients with resected hepatic colorectal cancer metastases. The greater the FGA, the more likely the patient is to survive. PMID- 15756003 TI - Expression profile of cancer-testis genes in 121 human colorectal cancer tissue and adjacent normal tissue. AB - PURPOSE: Among tumor antigens identified to date, cancer-testis (CT) antigens, which are coded by CT genes, are identified as a group of highly attractive targets for cancer vaccines. This study is the first to analyze the mRNA expression and possible correlation with pathologic characteristics of multiple CT genes in a large cohort of colorectal cancer (CRC) patients. EXPERIMENTAL DESIGN: The expression of 10 individual CT genes in 121 CRC and adjacent tissues were analyzed by RT-PCR method. The presence of autologous antibodies against NY ESO-1 was examined in serum samples by ELISA. To confirm the protein expression, immunohistochemistry was done for detecting the NY-ESO-1 antigen in mRNA-positive CRC tissues. RESULTS: The CT genes were detected with various frequencies in CRC tissue, SCP-1, 1.7%; SSX-2, 2.5%; SSX-4, 2.5%; SSX-1, 5.0%; CT10, 6.6%; NY-ESO-1, 9.9%; MAGE-1, 11.6%; LAGE-1, 15.7%; MAGE-4, 22.3%; and MAGE-3, 27.3%. In 56.2% of tumor tissues examined in this study, at least one CT gene was detected. In contrast, no CT gene expression was found in cancer adjacent tissues. Among 10 CT genes investigated, NY-ESO-1 and LAGE-1 are of particular interest because their mRNA expression in CRC was rarely reported before. In our study, NY-ESO-1 mRNA was found to express in 9.9% of the samples, and also correlated significantly with stages (P = 0.041) and local lymph node metastasis (P = 0.002). In addition, we also identified one NY-ESO-1 antibody-positive serum sample. MAGE-4 mRNA was expressed at a high frequency in tumor tissues with vessel emboli samples (P = 0.025). CONCLUSIONS: These results suggested that CT genes, especially NY-ESO-1 and LAGE-1, do express in CRC. More than 50% of the CRC patients in this study express at least one CT gene, making them eligible for CT vaccination. NY-ESO-1 gene may serve as a marker for local metastasis and advanced disease. MAGE-4 gene is significantly associated with the vessel emboli. PMID- 15756004 TI - The prognostic significance of plasma interleukin-6 levels in patients with metastatic hormone-refractory prostate cancer: results from cancer and leukemia group B 9480. AB - Interleukin-6 signaling can activate androgen receptor in a ligand-independent manner and may play an important functional role in hormone-refractory prostate cancer (HRCaP) progression and patient survival. Plasma and serum IL-6 levels have been associated with prostate cancer progression in several small studies. In order to evaluate its prognostic significance in metastatic HRCaP patients, we measured IL-6 in plasma collected at baseline from patients in a large cooperative group study [Cancer and Leukemia Group B 9480 (CALGB 9480)]. METHODS: 191 patients entered on CALGB 9480 had pretreatment plasma collected and centrally stored. Using a human IL-6 immunoassay, quantitative levels of IL-6 were measured in duplicate on 300 muL samples. The proportional hazard model was used to assess the prognostic significance of IL-6 in predicting overall survival. RESULTS: Median IL-6 level for the cohort of 191 patients was 4.80 pg/mL. Survival time among patients with IL-6 levels less than or equal to the median was 19 months (95% CI, 17-22) compared with 11 (95% CI, 8-14) months for patients above the median (P = 0.0004). In multivariate analysis, adjusting on performance status, lactate dehydrogenase, and prostate-specific antigen level, the hazard ratio was 1.38 (95% CI, 1.01-1.89; P = 0.043) using the median level as a cut point. Furthermore, a cut point of 13.31 pg/mL revealed robust prognostic significance with a hazard ratio of 2.02 (95% CI, 1.36-2.98; P = 0.0005). CONCLUSIONS: Plasma IL-6 level has prognostic significance in patients with metastatic HRCaP from CALGB 9480. These findings support using IL-6 levels in prognostic models and support the rationale for IL-6-targeted therapy in patients with HRCaP. PMID- 15756005 TI - Heterogeneous patterns of CEBPalpha mutation status in the progression of myelodysplastic syndrome and chronic myelomonocytic leukemia to acute myelogenous leukemia. AB - PURPOSE: We aimed to assess the role of CEBPalpha mutations in the progression of myelodysplastic syndrome (MDS) to acute myelogenous leukemia (AML) and their cooperating mutations. EXPERIMENTAL DESIGN: Mutational analysis of CEBPalpha with direct sequencing for each PCR product was done on matched bone marrow samples obtained from 50 adult patients with MDS at diagnosis and at AML transformation. Cloning analysis was used to determine the allelic distribution. RESULTS: CEBPalpha mutations were identified in four patients at diagnosis of MDS, including one with refractory anemia with excess blasts and three with chronic myelomonocytic leukemia. At AML transformation, three patients retained the identical mutant clones as their initial diagnosis, three acquired the mutations, and one lost CEBPalpha mutation when she gained FLT3/ITD mutation. Together, seven patients had CEBPalpha mutations throughout the disease course; four patients had NH(2)-terminal mutations resulting in a frameshift and truncation of the protein, three of them had two different mutations either on the same alleles or on different alleles, two had missense mutations, and one had a deletion in the basic region leucine zipper domain. Except for one with coexistence of N-ras mutation, no sample harbored cooperating mutations with FLT3 or N-ras genes. CEBPalpha mutations had no influence on the time to AML progression or overall survival. CONCLUSIONS: Our results show that CEBPalpha mutations play a role in a subset of patients with MDS, especially in chronic myelomonocytic leukemia. The mutation status was heterogeneous, exhibiting identical clone, clonal change, or clonal evolution during the progression to AML. PMID- 15756006 TI - The clinical significance of Aurora-A/STK15/BTAK expression in human esophageal squamous cell carcinoma. AB - PURPOSE: Aurora-A/STK15/BTAK (Aurora-A) encodes a Serine/Threonine kinase associated with chromosomal distribution, and its up-regulation induces chromosomal instability thereby leading to aneuploidy and cell transformation in several types of cancer. In this study, we investigated the role of Aurora-A in human esophageal squamous cell carcinoma (ESCC). EXPERIMENTAL DESIGN: The expression levels of Aurora-A mRNA were compared in 33 ESCC tissues with that in corresponding normal esophageal epithelium by semiquantitative reverse transcription-PCR, and the distribution patterns and expression levels of Aurora A protein were immunohistochemically investigated in the ESCC tumors of 142 patients. The results were then separately compared with the clinicopathologic findings of the patients, and the expression of Aurora-A was examined in nine ESCC cell lines and a normal esophageal epithelial cell line using Western blot analysis. RESULTS: The up-regulation of Aurora-A mRNA was found in 30% (10 of 33) of the tumors by semiquantitative reverse transcription-PCR, and protein up regulation was found in 53% (75 of 142) of the patients by immunohistochemistry. mRNA and protein up-regulation of Aurora-A were correlated with distant lymph node metastasis (P = 0.05 and P = 0.04, respectively), and patients with Aurora-A mRNA or protein up-regulation had a poorer prognosis (P = 0.003 and P = 0.0009, respectively). Furthermore, multivariate analysis revealed that up-regulation of the Aurora-A protein was an independent prognostic factor. In addition, Aurora-A expression in all ESCC cell lines was higher than that in a normal esophageal epithelial cell line. CONCLUSIONS: The up-regulation of Aurora-A expression may reflect the malignant behavior of ESCC and may prove useful information as a prognostic factor for ESCC patients. PMID- 15756007 TI - Expression of CXCR4 predicts poor prognosis in patients with malignant melanoma. AB - PURPOSE: CXCR4 receptor and its unique ligand, the CXCL12 chemokine, have been recently implicated in cancer metastasis. Evidence about the role of CXCR4/CXCL12 axis has been reported in several cancers including melanoma. Our goal was to investigate if CXCR4 expression has a prognostic value in malignant melanoma. EXPERIMENTAL DESIGN: Immunohistochemical expression of CXCR4 was evaluated on 71 specimens of primary cutaneous melanoma with a Breslow tumor thickness of >1 mm after radical resection. Associations between baseline patient features and tumors were analyzed by chi(2) test. The prognostic value of CXCR4 expression was evaluated by univariate and multivariate analyses adjusted by age, sex, Breslow tumor thickness, presence of ulceration, and sentinel lymph node metastases. RESULTS: CXCR4 expression was detected in 31 of 71 (43.6%) primary cutaneous melanomas. Membrane or cytoplasmic staining for CXCR4 protein was absent in 56% of the tumors. The positive cases were divided into three score classes according to their staining: low in 15 cases (21%), moderate in 10 (14%), and high in 6 (8%). After a median follow-up of 38 months, 26 patients progressed (16 of 26 expressed CXCR4) and 19 died (12 of 19 expressed CXCR4). The CXCR4 expression on tumor cells was correlated with an unfavorable prognosis with a median disease free and overall survival of 22 and 35 months, respectively. The hazard ratios of relapse and death, compared with patients with CXCR4-negative tumors, were 2.5 (95% confidence interval, 1.2-6.1) and 3.1 (95% confidence interval, 1.1-7.2), respectively. Median time-to-event (progression and survival) was not reached in patients with CXCR4-negative tumors. In the multivariate analysis, CXCR4 expression, presence of ulceration, and sentinel lymph node status emerged as independent prognostic factors. CONCLUSIONS: This article provides the first evidence that CXCR4 expression could be an independent and powerful prognostic marker in primary cutaneous malignant melanomas. PMID- 15756008 TI - B7-h4 is highly expressed in ductal and lobular breast cancer. AB - PURPOSE: This study was designed to investigate the expression of B7-H4 protein, a member of the B7 family that is involved in the regulation of antigen-specific immune responses, in normal breast and in primary and metastatic breast carcinomas. EXPERIMENTAL DESIGN: Archival formalin-fixed tissue blocks from breast cancers and normal somatic tissues were evaluated for B7-H4 expression by immunohistochemistry with manual and automated image analysis. The proportion of B7-H4-positive cells and the intensity of B7-H4 staining were compared with histologic type, grade, stage, hormone receptor status, and HER-2/neu status. RESULTS: B7-H4 was detected in 165 of 173 (95.4%) primary breast cancers and in 240 of 246 (97.6%) metastatic breast cancers. B7-H4 staining intensity was greater in invasive ductal carcinomas [24.61 relative units (RU)] and in invasive lobular carcinomas (15.23 RU) than in normal breast epithelium (4.30 RU, P = 0.0003). Increased staining intensity was associated with negative progesterone receptor status (P = 0.014) and history of neoadjuvant chemotherapy (P = 0.004), and the proportion of B7-H4-positive cells was associated with negative progesterone receptor (P = 0.001) and negative HER-2/neu (P = 0.024) status. However, there was no statistically significant relationship between the proportion of B7-H4-positive cells or staining intensity and grade, stage, or other clinicopathologic variables. Low levels of B7-H4 expression were also detected in epithelial cells of the female genital tract, lung, pancreas, and kidney, but B7-H4 was generally absent in most other normal somatic tissues. CONCLUSIONS: The nearly ubiquitous expression of B7-H4 in breast cancer, independent of tumor grade or stage, suggests a critical role for this protein in breast cancer biology. PMID- 15756009 TI - NBS1 expression as a prognostic marker in uveal melanoma. AB - PURPOSE: Up to half of uveal melanoma patients die of metastatic disease. Treatment of the primary eye tumor does not improve survival in high-risk patients due to occult micrometastatic disease, which is present at the time of eye tumor diagnosis but is not detected and treated until months to years later. Here, we use microarray gene expression data to identify a new prognostic marker. EXPERIMENTAL DESIGN: Microarray gene expression profiles were analyzed in 25 primary uveal melanomas. Tumors were ranked by support vector machine (SVM) and by cytologic severity. Nbs1 protein expression was assessed by quantitative immunohistochemistry in 49 primary uveal melanomas. Survival was assessed using Kaplan-Meier life-table analysis. RESULTS: Expression of the Nijmegen breakage syndrome (NBS1) gene correlated strongly with SVM and cytologic tumor rankings (P < 0.0001). Further, immunohistochemistry expression of the Nbs1 protein correlated strongly with both SVM and cytologic rankings (P < 0.0001). The 6-year actuarial survival was 100% in patients with low immunohistochemistry expression of Nbs1 and 22% in those with high Nbs1 expression (P = 0.01). CONCLUSIONS: NBS1 is a strong predictor of uveal melanoma survival and potentially could be used as a clinical marker for guiding clinical management. PMID- 15756011 TI - Induction of p53 up-regulated modulator of apoptosis messenger RNA by chemotherapeutic treatment of locally advanced breast cancer. AB - PURPOSE: In biopsies of patients with locally advanced breast cancer, we investigated the in vivo changes of the gene expression pattern induced by chemotherapy to find genes that are potentially responsible for the efficacy of the drug. EXPERIMENTAL DESIGN: Early cellular responses to chemotherapy-induced damage, both in vivo and in vitro, were investigated by analyzing chemotherapy induced changes in gene expression profiles. Core biopsies were taken from nine patients with locally advanced breast cancer, before and at 6 hours after initiation of doxorubicin-based chemotherapy. Both samples were cohybridized on the same microarray containing 18,000 cDNA spots. RESULTS: The analysis revealed marked differences in gene expression profile between treated and untreated samples. The gene which was most frequently found to be differentially expressed was p53 up-regulated modulator of apoptosis (PUMA). This gene was up-regulated in eight of nine patients with an average factor of 1.80 (range, 1.36-2.73). In vitro MCF-7 breast cancer cells exposed to clinically achievable doxorubicin concentrations for 6 hours revealed marked induction of PUMA mRNA, as well. CONCLUSIONS: This is the first report describing PUMA mRNA to be up-regulated as a response to chemotherapy in patients. Because PUMA is a known member of the family of BH3-only proapoptotic proteins, this finding suggests PUMA's potential importance for the response to anticancer drugs. PMID- 15756010 TI - Phase I clinical and pharmacokinetic study of kahalalide F in patients with advanced androgen refractory prostate cancer. AB - PURPOSE: The purpose is to determine the maximum tolerated dose, profile of adverse events, and dose-limiting toxicity of Kahalalide F (KF) in patients with androgen refractory prostate cancer. Furthermore, the pharmacokinetics after KF administration and preliminary antitumor activity were evaluated. KF is a dehydroaminobutyric acid-containing peptide isolated from the marine herbivorous mollusk, Elysia rufescens. EXPERIMENTAL DESIGN: Adult patients with advanced or metastatic androgen refractory prostate cancer received KF as an i.v. infusion over 1 hour, during five consecutive days every 3 weeks. The starting dose was 20 microg per m(2) per day. Clinical pharmacokinetics studies were done in all patients using noncompartmental analysis. Prostate-specific antigen levels were evaluated as a surrogate marker for activity against prostate cancer. RESULTS: Thirty-two patients were treated at nine dose levels (20-930 microg per m(2) per day). The maximum tolerated dose on this schedule was 930 microg per m(2) per day. The dose-limiting toxicity was reversible and asymptomatic Common Toxicity Criteria grade 3 and 4 increases in transaminases. The recommended dose for phase II studies is 560 microg per m(2) per day. Pharmacokinetics analysis revealed dose linearity up to the recommended dose. Thereafter, a more than proportional increase was observed. Elimination was rapid with a mean (SD) terminal half-life (t(1/2)) of 0.47 hour (0.11 hour). One patient at dose level 80 microg per m(2) per day had a partial response with a prostate-specific antigen decline by at least 50% for > or =4 weeks. Five patients showed stable disease. CONCLUSIONS: KF can be given safely as a 1-hour i.v. infusion during five consecutive days at a dose of 560 microg per m(2) per day once every 3 weeks. PMID- 15756012 TI - Phase II evaluation of docetaxel-modulated capecitabine in previously treated patients with non-small cell lung cancer. AB - PURPOSE: Based on the preclinical observation of upregulation of thymidine phosphorylase, the last enzymatic step in the conversion of capecitabine to 5 fluorouracil, by docetaxel along with good clinical tolerability of the combination of docetaxel and capecitabine using an optimized schedule in a previous phase I trial, we conducted this phase II study of this combination in patients with refractory or relapsed non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with NSCLC previously treated with at least one platinum- or paclitaxel-based regimen received docetaxel 36 mg/m(2) on days 1, 8, and 15 and capecitabine 625 mg/m(2) twice daily on days 5 to 18, every 4 weeks. The primary objective of the study was evaluation of progression-free survival (PFS) 26 weeks from initiation of treatment. RESULTS: Thirty-six evaluable patients received 104 cycles of the combination. Severe toxicities were infrequent with only one patient requiring toxicity-related hospitalization. The 26-week PFS rate was 25% (95% confidence interval, 12-42) with an intent to treat median survival and 1-year survival rate of 9.1 months and 37%, respectively. Among 31 patients with measurable disease (Response Evaluation Criteria in Solid Tumors criteria), eight (26%; 95% confidence interval, 12-45) achieved partial responses. CONCLUSION: The combination of capecitabine and weekly docetaxel is well tolerated in previously treated patients with NSCLC. The relatively high 26-week PFS and 1-year survival, as well as the high response rate observed, encourages further evaluation of this regimen in NSCLC, either in randomized trials for refractory patients or as a potential treatment option for chemotherapy naive patients. PMID- 15756014 TI - Phase I trial and pharmacokinetic study of raltitrexed in children with recurrent or refractory leukemia: a pediatric oncology group study. AB - PURPOSE: To evaluate the toxicity, antileukemic activity, and pharmacology of raltitrexed administered weekly for 3 weeks to patients with refractory or recurrent leukemia. EXPERIMENTAL DESIGN: Raltitrexed was administered as a 15 minute infusion for 3 consecutive weeks every 5 weeks, at doses ranging from 1.3 to 2.8 mg/m(2). The first course was used to determine the dose-limiting toxicities and maximum tolerated dose. Correlative studies included an assessment of raltitrexed pharmacokinetics and measurement of plasma 2'-deoxyuridine concentrations, a surrogate measure of thymidylate synthase inhibition. RESULTS: Twenty-one children (18 evaluable) with refractory leukemia received 25 courses of raltitrexed. The dose-limiting toxicity was reversible elevation in liver transaminases at the 2.8-mg/m(2) dose level and the maximum tolerated dose was 2.1 mg/m(2) per dose. Pharmacokinetics were best characterized by a two compartment model with a clearance of 139 mL/min/m(2) (8.3 L/h/m(2)), a 2.4-L volume of distribution, an initial half-life (t(1/2alpha)) of 6 minutes, and a terminal half-life (t(1/2beta)) of 45 minutes. There were three objective responses. CONCLUSIONS: Raltitrexed was well tolerated when administered as a single agent to children with recurrent or refractory leukemia. We observed preliminary evidence of antileukemia activity using this weekly dosing schedule and these observations support further evaluation of raltitrexed in this population. PMID- 15756013 TI - A phase I trial of the novel farnesyl protein transferase inhibitor, BMS-214662, in combination with paclitaxel and carboplatin in patients with advanced cancer. AB - PURPOSE: This phase I study was conducted to determine the toxicities, pharmacokinetics, and pharmacodynamics of BMS-214662, a farnesyl transferase inhibitor, in combination with paclitaxel and carboplatin, in patients with advanced solid tumors. EXPERIMENTAL DESIGN: Patients with solid tumors received one of six escalating dose levels of BMS-214662 infused over 1 hour given following paclitaxel and carboplatin on the first day of a 21-day cycle. Toxicities were graded by the National Cancer Institute common toxicity criteria and recorded as maximum grade per patient for each treatment cycle. Inhibition of farnesyl transferase activity in peripheral blood mononuclear cells (PBMCs) was evaluated. Accumulation of unfarnesylated HDJ-2 in PBMCs of patients was evaluated as a marker of farnesyl transferase inhibition by BMS-214662. RESULTS: Thirty patients received 141 cycles of treatment through six dose levels. Dose limiting toxicities were neutropenia, thrombocytopenia, nausea, and vomiting. There was no pharmacokinetic interaction between BMS-214662 and paclitaxel. The maximum tolerated dose was established as BMS-214662 (160 mg/m(2)), paclitaxel (225 mg/m(2)) and carboplatin (area under the curve = 6 on day 1), every 21 days. Inhibition of HDJ-2 farnesylation in PBMCs of patients was shown. One measurable partial response was observed in a patient with taxane-resistant esophageal cancer. There was partial regression of evaluable disease in two other patients (endometrial and ovarian cancer). Stable disease (> 4 cycles) occurred in eight other patients. CONCLUSIONS: The combination of BMS-214662 with paclitaxel and carboplatin was well tolerated, with broad activity in solid tumors. There was no correlation between dose level and accumulation of unfarnesylated HDJ-2 in PBMCs nor tumor response. PMID- 15756015 TI - A randomized, double-blinded, placebo-controlled phase II trial of recombinant human leukemia inhibitory factor (rhuLIF, emfilermin, AM424) to prevent chemotherapy-induced peripheral neuropathy. AB - PURPOSE: To determine whether recombinant human leukemia inhibitory factor (rhuLIF, AM424, emfilermin) can prevent or ameliorate the development of chemotherapy-induced peripheral neuropathy (CIPN) after treatment with carboplatin (AUC 6) and paclitaxel (175 mg/m(2) over 3 hours). EXPERIMENTAL DESIGN: Randomized double-blind placebo-controlled phase II clinical trial. Eligible patients had solid tumors for which treatment with carboplatin/paclitaxel was appropriate. The primary end point was a standardized composite peripheral nerve electrophysiology (CPNE) score, based on nerve velocities and amplitudes, measured at baseline and after four cycles of chemotherapy. Secondary efficacy end points included CPNE score at last cycle and at exit evaluation, vibration perception threshold, H-reflex latency, symptom scores, and quantitative assessment of neurologic signs. Study drug was given s.c. daily for 7 days starting the day before chemotherapy. Patients were randomized to receive low-dose rhuLIF (2 microg/kg), high-dose rhuLIF (4 microg/kg), or placebo. RESULTS: Patients (n = 117) were randomized across seven neurology test centers. Thirty-six patients received low dose rhuLIF (2 microg/kg), 39 received high dose rhuLIF (4 microg/kg) and 42 received placebo. rhuLIF was well tolerated with 95% compliance and no adverse effects on quality of life. No differences between groups in CPNE or any of the individual neurologic testing variables were observed between baseline and cycle 4 or by the secondary efficacy variables. CONCLUSIONS: rhuLIF is not effective in preventing CIPN caused by carboplatin and paclitaxel. CPNE is a reliable and valid tool that was sensitive to the onset and progression of CIPN. PMID- 15756016 TI - Intratumoral recombinant human interleukin-12 administration in head and neck squamous cell carcinoma patients modifies locoregional lymph node architecture and induces natural killer cell infiltration in the primary tumor. AB - The objective of this study was to evaluate the histologic and immunohistopathologic effects of intratumorally given recombinant human interleukin-12 on the immune cells in the primary tumors and regional lymph nodes. Ten previously untreated patients with head and neck squamous cell carcinoma (HNSCC) were injected in the primary tumor twice to thrice, once weekly, at two dose levels of 100 or 300 ng/kg, before surgery. These patients were compared with 20 non-IL-12-treated control HNSCC patients. In the primary tumor, the number of CD56+ natural killer (NK) cells was increased in IL-12 treated patients compared with control patients. In some IL-12-treated patients, an impressive peritumoral invasion of CD20+ B cells was noticed. No differences were seen in the CD8+ or CD4+ T lymphocytes. Interestingly, major differences were apparent in the architecture of the enlarged lymph nodes of IL-12-treated patients; in particular, the distribution of B cells differed and fewer primary and secondary follicles with smaller germinal centers were observed. In addition, a decrease of dendritic cell lysosyme-associated membrane glycoprotein-positive cells in the paracortex was noted, resulting in a reduction of paracortical hyperplasia. In the lymph nodes, especially the CD56+ NK cells but also the CD8+ and CD4+ T lymphocytes, produced a high amount of IFN-gamma. Patients, irrespectively of IL-12 treatment, with a high number of CD56+ cells in the primary tumor had a better overall survival than those with a low number. In conclusion, after i.t. IL-12 treatment in HNSCC patients, the largest effect was seen on the NK cells, with a higher number in the primary tumor and a high IFN gamma mRNA expression in the lymph nodes. Significant effects were noted on B cells, with altered lymph node architecture in every IL-12-treated patient and excessive peritumoral infiltration in some patients. PMID- 15756017 TI - A phase I study of alpha-galactosylceramide (KRN7000)-pulsed dendritic cells in patients with advanced and recurrent non-small cell lung cancer. AB - PURPOSE: Human Valpha24 natural killer T (NKT) cells bearing an invariant Valpha24JalphaQ antigen receptor, the counterpart of murine Valpha14 NKT cells, are activated by a specific ligand, alpha-galactosylceramide (alphaGalCer, KRN7000), in a CD1d-dependent manner. I.v. administration of alphaGalCer-pulsed dendritic cells (DC) induces significant activation and expansion of Valpha14 NKT cells in the lung and resulting potent antitumor activities in mouse tumor metastatic models. We did a phase I dose escalation study with alphaGalCer-pulsed DCs in lung cancer patients. EXPERIMENTAL DESIGN: Patients with advanced non small cell lung cancer or recurrent lung cancer received i.v. injections of alphaGalCer-pulsed DCs (level 1: 5 x 10(7)/m(2); level 2: 2.5 x 10(8)/m(2); and level 3: 1 x 10(9)/m(2)) to test the safety, feasibility, and clinical response. Immunomonitoring was also done in all completed cases. RESULTS: Eleven patients were enrolled in this study. No severe adverse events were observed during this study in any patient. After the first and second injection of alphaGalCer-pulsed DCs, dramatic increase in peripheral blood Valpha24 NKT cells was observed in one case and significant responses were seen in two cases receiving the level 3 dose. No patient was found to meet the criteria for partial or complete responses, whereas two cases in the level 3 group remained unchanged for more than a year with good quality of life. CONCLUSIONS: In this clinical trial, alphaGalCer pulsed DC administration was well tolerated and could be safely done even in patients with advanced disease. PMID- 15756018 TI - Growth inhibitory and antimetastatic effect of green tea polyphenols on metastasis-specific mouse mammary carcinoma 4T1 cells in vitro and in vivo systems. AB - PURPOSE: Breast cancer is the second leading cause of cancer-related deaths among females. Dietary habits may have a role in breast cancer risk and prevention as well. Here, we examined the effect of green tea polyphenols (GTP) on growth and metastasis of highly metastatic mouse mammary carcinoma 4T1 cells in vitro and in vivo systems. EXPERIMENTAL DESIGN: 4T1 cells were treated with (-) epigallocatechin-3-gallate (EGCG), and the effect was determined on cellular proliferation, induction of apoptosis, proapoptosis, and antiapoptotic proteins of Bcl-2 family, and caspase 3 and poly(ADP-ribose) polymerase activation following 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and Western blot analysis. Anticarcinogenic and antimetastatic effect of GTP in 4T1 cells was assessed in immunocompetent BALB/c mice. RESULTS: Treatment of 4T1 cells with EGCG resulted in inhibition of cell proliferation, induction of apoptosis in dose- and time-dependent manner. The increase in apoptosis was accompanied with decrease in the protein expression of Bcl-2 concomitantly increase in Bax, cytochrome c release, Apaf-1, and cleavage of caspase 3 and PARP proteins. Treatment of EGCG-rich GTP in drinking water to 4T1 cells bearing BALB/c mice resulted in reduction of tumor growth accompanied with increase in Bax/Bcl-2 ratio, reduction in proliferating cell nuclear antigen and activation of caspase 3 in tumors. Metastasis of tumor cells to lungs was inhibited and survival period of animals was increased after green tea treatment. CONCLUSION: This study suggests that GTP have the ability to prevent the development of breast cancer and its metastasis; however, further in vivo studies are required to identify the molecular targets. PMID- 15756019 TI - Effects of imatinib on monocyte-derived dendritic cells are mediated by inhibition of nuclear factor-kappaB and Akt signaling pathways. AB - Dendritic cells are the most powerful antigen-presenting cells playing a decisive role for the initiation and maintenance of primary immune responses. However, signaling pathways involved in the differentiation of these cells have not been fully determined. Imatinib is a novel tyrosine kinase inhibitor effective against Abl kinases, c-Kit, and platelet-derived growth factor receptor. Using this compound, we show that human monocyte-derived dendritic cells generated in the presence of therapeutic concentrations of imatinib show a reduced expression of CD1a, MHC class I and II, and costimulatory molecules as well as decreased secretion of chemokines and cytokines resulting in an impaired capacity of dendritic cells to elicit primary T-cell responses. Using Western blot analyses, we found that these effects are mediated by inhibition of phosphatidylinositol 3 kinase/Akt pathways and a pronounced down-regulation of nuclear localized protein levels of nuclear factor-kappaB family members. Importantly, using blocking antibodies and tyrosine kinase inhibitors, we show that the inhibitory effects of imatinib on dendritic cell differentiation are not mediated via platelet-derived growth factor receptor and c-Kit. Taken together, our study reveals that imatinib inhibits dendritic cell differentiation and function via Akt and nuclear factor kappaB signal transduction. Importantly, we show that imatinib can inhibit the function of normal, nonmalignant cells that may result in immunosuppression of these patients. PMID- 15756020 TI - Cure of mammary carcinomas in Her-2 transgenic mice through sequential stimulation of innate (neoadjuvant interleukin-12) and adaptive (DNA vaccine electroporation) immunity. AB - PURPOSE: Whereas neoadjuvant therapy is emerging as a treatment option in early primary breast cancer, no data are available on the use of antiangiogenic and immunomodulatory agents in a neoadjuvant setting. In a model of Her-2 spontaneous mammary cancer, we investigated the efficacy of neoadjuvant interleukin 12 (IL 12) followed by "immune-surgery" of the residual tumor. EXPERIMENTAL DESIGN: Female BALB/c mice transgenic for the rat Her-2 oncogene inexorably develop invasive carcinomas in all their mammary glands by the 23rd week of age. Mice with multifocal in situ carcinomas received four weekly i.p. injections of 100 ng IL-12 followed by a 3-week rest. This course was given four times. A few mice additionally received DNA plasmids encoding portions of the Her-2 receptor electroporated through transcutaneous electric pulses. RESULTS: The protection elicited by IL-12 in combination with two DNA vaccine electroporations kept 63% of mice tumor-free. Complete protection of all 1-year-old mice was achieved when IL-12-treated mice received four vaccine electroporations. Pathologic findings, in vitro tests, and the results from immunization of both IFN-gamma and immunoglobulin gene knockout transgenic mice and of adoptive transfer experiments all show that IL-12 augments the B- and T-cell response elicited by vaccination and slightly decreases the number of regulatory T cells. In addition, IL-12 strongly inhibits tumor angiogenesis. CONCLUSIONS: In Her-2 transgenic mice, IL 12 impairs tumor progression and triggers innate immunity so markedly that DNA vaccination becomes effective at late points in time when it is ineffective on its own. PMID- 15756021 TI - Triterpenoid electrophiles (avicins) suppress heat shock protein-70 and x-linked inhibitor of apoptosis proteins in malignant cells by activation of ubiquitin machinery: implications for proapoptotic activity. AB - Avicins are plant-derived triterpenoid stress metabolites that have both proapoptotic and cytoprotective properties. Avicins induce apoptosis in Jurkat T leukemia cells by targeting mitochondria and release of cytochrome c that occurs in a p53-independent manner. However, postmitochondrial antiapoptotic barriers, such as increased expression of heat shock proteins (Hsp) and X-linked inhibitor of apoptosis proteins (XIAP), frequently exist in cancer cells and often account for resistance to chemotherapy and a poor prognosis. In this article, we show the role of avicins in the activation of stress-regulated ubiquitination and degradation of Hsp70 and XIAP. This is the first report showing the regulation of Hsp70 via the ubiquitin/proteasome pathway. We also show the induction of E3alpha ubiquitin ligase in avicin-treated Jurkat T leukemia cells, and its involvement in the degradation of XIAP. Avicin-mediated suppression of Hsp70 and XIAP was further confirmed in other leukemic/lymphoma cell lines and freshly isolated peripheral blood lymphocytes from Sezary syndrome patients. No change in the Hsp70 and XIAP proteins was observed in peripheral blood lymphocytes from normal donors. We propose that the ability of avicins to induce ubiquitination and regulate the degradation of Hsp70 and XIAP in leukemia cells could have important implications in the treatment of drug-resistant neoplasia and inflammatory disorders. PMID- 15756022 TI - AEE788, a dual tyrosine kinase receptor inhibitor, induces endothelial cell apoptosis in human cutaneous squamous cell carcinoma xenografts in nude mice. AB - PURPOSE: We investigated whether concomitant blockade of the epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor (VEGFR) signaling pathways by AEE788, a dual inhibitor of EGFR and VEGFR tyrosine kinases, would inhibit the growth of cutaneous squamous cell carcinoma (SCC) cells and human cutaneous cancer xenografts in nude mice. EXPERIMENTAL DESIGN: We examined the effects of AEE788 on the phosphorylation of EGFR and VEGFR-2 in cutaneous SCC cells expressing EGFR and VEGFR-2 and cutaneous SCC cell growth and apoptosis. We assessed the in vivo antitumor effects of AEE788 in a xenograft model in nude mice. AEE788 (50 mg/kg) was given orally thrice weekly to mice that had been s.c. injected with Colo16 tumor cells. Mechanisms of in vivo AEE788 activity were determined by immunohistochemical analysis. RESULTS: Treatment of cutaneous SCC cells with AEE788 led to dose-dependent inhibition of EGFR and VEGFR-2 phosphorylation, growth inhibition, and induction of apoptosis. In mice treated with AEE788, tumor growth was inhibited by 54% at 21 days after the start of treatment compared with control mice (P < 0.01). Immunohistochemical analysis revealed that AEE788 inhibited phosphorylation of EGFR and VEGFR and induced apoptosis of tumor cells and tumor-associated endothelial cells. CONCLUSIONS: In addition to inhibiting cutaneous cancer cell growth by blocking EGFR and VEGFR signaling pathways in vitro, AEE788 inhibited in vivo tumor growth by inducing tumor and endothelial cell apoptosis. PMID- 15756023 TI - Growth inhibition of multiple myeloma cells by a novel IkappaB kinase inhibitor. AB - Involvement of nuclear factor-kappaB (NF-kappaB) in cell survival and proliferation of multiple myeloma has been well established. In this study we observed that NF-kappaB is constitutively activated in all human myeloma cell lines, thus confirming the previous studies. In addition, we found the phosphorylation of p65 subunit of NF-kappaB in addition to the phosphorylation of IkappaBalpha and the activation of NF-kappaB DNA binding and that various target genes of NF-kappaB including bcl-x(L), XIAP, c-IAP1, cyclin D1, and IL-6 are up regulated. We then examined the effect of a novel IkappaB kinase inhibitor, 2 amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-piperidin-4-yl nicotinonitrile (ACHP). When myeloma cells were treated with ACHP, the cell growth was efficiently inhibited with IC(50) values ranging from 18 to 35 mumol/L concomitantly with inhibition of the phosphorylation of IkappaBalpha/p65 and NF kappaB DNA-binding, down-regulation of the NF-kappaB target genes, and induction of apoptosis. In addition, we observed the treatment of ACHP augmented the cytotoxic effects of vincristine and melphalan (l-phenylalanine mustard), conventional antimyeloma drugs. These findings indicate that IkappaB kinase inhibitors such as ACHP can sensitize myeloma cells to the cytotoxic effects of chemotherapeutic agents by blocking the antiapoptotic nature of myeloma cells endowed by the constitutive activation of NF-kappaB. PMID- 15756024 TI - Pulsatile administration of the epidermal growth factor receptor inhibitor gefitinib is significantly more effective than continuous dosing for sensitizing tumors to paclitaxel. AB - PURPOSE: Gefitinib is an inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase. Continuous inhibition of EGFR signaling is thought necessary for optimal inhibition of tumor cell proliferation. We hypothesized that continuous gefitinib may antagonize the effects of cytotoxics that inhibit tumor cells in other phases of the cell cycle. Furthermore, we hypothesized that intermittent dosing would allow for dose escalation and greater inhibition of EGFR-dependent antiapoptotic pathways. EXPERIMENTAL DESIGN: To test these assertions, we compared combinations of paclitaxel and gefitinib using either intermittent or continuous dosing schedules in mice. RESULTS: We found that when used in combination with paclitaxel, pulsatile gefitinib was significantly superior to continuous dosing. When gefitinib was administered for one or two consecutive days before paclitaxel, much higher doses could be given safely. Two days of gefitinib treatment before paclitaxel was most effective, causing significantly greater mean tumor regression and a higher percentage of complete responses than other schedules. CONCLUSIONS: The results suggest that the dose and schedule of an EGFR inhibitor required to effectively inhibit proliferation may differ from that required to stimulate apoptosis or to induce other effects. The former may require continuous EGFR inhibition to maintain cell cycle arrest, whereas sensitization to apoptosis may be optimally induced by profound but temporary inhibition of survival pathways. Our data suggest that the effects of receptor inhibition vary as a function of dose and schedule and that continuous administration of tyrosine kinase inhibitors may not be the best schedule with which to combine such agents with taxanes. PMID- 15756025 TI - Antitumor efficacy of bcl-2 and c-myc antisense oligonucleotides in combination with cisplatin in human melanoma xenografts: relevance of the administration sequence. AB - PURPOSE: bcl-2 and c-myc oncogenes are frequently overexpressed in different human tumors, including melanoma. Here, we evaluate the combined efficacy of two antisense oligonucleotides targeting bcl-2 mRNA (ODN bcl-2) and c-myc mRNA (ODN c myc) in combination with cis-diammine dichloroplatinum (cisplatin, DDP) on three human melanoma lines (LM, NG, and M20). EXPERIMENTAL DESIGN: Two different sequences were designed to treat tumor-bearing mice: in the first one, ODN bcl-2 at a dose of 0.2 mg/day x4, followed by DDP given i.p. at a dose of 3.3 mg/kg/day x3 and ODN c-myc i.v. at 0.5 mg/day x7, whereas the other sequence consisted of ODN c-myc given as first agent followed by DDP and ODN bcl-2 at the same doses. Mice received three complete cycles of treatment in 1-week intervals. RESULTS: The treatment sequence with ODN bcl-2/DDP/ODN c-myc combination completely inhibited growth in NG tumor and induced a 35-day delay in LM tumor growth. In contrast, the M20 tumor growth was unaffected by the combination. A discrete amount of c-Myc and bcl-2 protein expression in both LM and NG tumors was detected, whereas no detectable levels of the two proteins were observed in M20 tumors. Compared with the other combination, the sequence (ODN bcl-2/DDP/ODN c myc) produced the most effective results, producing a significant decrease in bcl 2 and c-Myc protein expression, which in turn significantly increased the survival of NG- and LM-bearing mice, with 4 mice out of 11 and 1 out of 7 mice being cured, respectively. Finally, this combination increased the apoptotic rate and produced an antiangiogenetic effect. CONCLUSIONS: These results show that an antisense approach to the treatment of melanoma xenografts overexpressing either bcl-2 or c-myc oncogenes represents a successful strategy to improve the response to chemotherapy in melanoma, with particular attention to the treatment sequence. PMID- 15756026 TI - Celecoxib inhibits prostate cancer growth: evidence of a cyclooxygenase-2 independent mechanism. AB - PURPOSE: Selective cyclooxygenase-2 (COX-2) inhibitors may suppress carcinogenesis by both COX-2-dependent and COX-2-independent mechanisms. The primary purpose of this study was to evaluate whether celecoxib or rofecoxib, two widely used selective COX-2 inhibitors, possess COX-2-independent antitumor activity. EXPERIMENTAL DESIGN: PC3 and LNCaP human prostate cancer cell lines were used to investigate the growth inhibitory effects of selective COX-2 inhibitors in vitro. To complement these studies, we evaluated the effect of celecoxib on the growth of PC3 xenografts. RESULTS: COX-1 but not COX-2 was detected in PC3 and LNCaP cells. Clinically achievable concentrations (2.5-5.0 micromol/L) of celecoxib inhibited the growth of both cell lines in vitro, whereas rofecoxib had no effect over the same concentration range. Celecoxib inhibited cell growth by inducing a G(1) cell cycle block and reducing DNA synthesis. Treatment with celecoxib also led to dose-dependent inhibition of PC3 xenograft growth without causing a reduction in intratumor prostaglandin E(2). Inhibition of tumor growth occurred at concentrations (2.37-5.70 micromol/L) of celecoxib in plasma that were comparable with the concentrations required to inhibit cell growth in vitro. The highest dose of celecoxib led to a 52% reduction in tumor volume and an approximately 50% decrease in both cell proliferation and microvessel density. Treatment with celecoxib caused a marked decrease in amounts of cyclin D1 both in vitro and in vivo. CONCLUSIONS: Two clinically available selective COX-2 inhibitors possess different COX-2 independent anticancer properties. The anticancer activity of celecoxib may reflect COX-2-independent in addition to COX-2-dependent effects. PMID- 15756027 TI - Herceptin down-regulates HER-2/neu and vascular endothelial growth factor expression and enhances taxol-induced cytotoxicity of human Ewing's sarcoma cells in vitro and in vivo. AB - We have previously shown that high levels of HER-2/neu protein were overexpressed in human Ewing's sarcoma cells (TC71, SK-ES1) relative to normal human osteoblasts. The purpose of this study was to determine whether herceptin alone or in combination with chemotherapeutic agents could inhibit the growth of Ewing's sarcoma in vitro and in vivo. Western blot analysis showed that the protein levels of HER-2/neu were decreased following herceptin treatment. Cell growth was also inhibited by herceptin in a dose-dependent manner with an IC(50) of 4 mg/mL in TC71 and SK-ES1 cell line, whereas human immunoglobin had no effect. Northern blot and ELISA showed the RNA expression and protein levels of vascular endothelial growth factor were also inhibited by herceptin treatment with no alteration in HIF-1alpha protein and topoisomerase IIalpha expression. Furthermore, Ewing's sarcoma tumor growth was significantly delayed by 100 mg/kg herceptin treatment in our Ewing's sarcoma xenograft mouse model. Combining taxol with herceptin resulted in additive cytotoxicity, whereas herceptin-etoposide, doxorubicin, and 9-nitrocamptothecin combinations did not. Taxol-herceptin enhanced growth inhibition in TC71 cells in vitro compared with either agent alone. Ewing's sarcoma growth was also delayed in vivo and mean tumor size was significantly lower in mice treated with herceptin plus taxol than in those receiving taxol or herceptin alone. These data suggest that herceptin in combination with taxol may be a therapeutic option in the treatment of Ewing's sarcoma. PMID- 15756028 TI - Effect of polyethylene glycol linker chain length of folate-linked microemulsions loading aclacinomycin A on targeting ability and antitumor effect in vitro and in vivo. AB - PURPOSE: To establish a novel formulation tumor-targeted drug carrier of lipophilic antitumor antibiotics, aclacinomycin A (ACM), folate-linked microemulsions were prepared and investigated both in vitro and in vivo. EXPERIMENTAL DESIGN: Three kinds of folate-linked microemulsions with different polyethylene glycol (PEG) chain lengths loading ACM were formulated with 0.24 mol% folate-PEG(2000)-distearoylphosphatidylethanolamine (DSPE), folate-PEG(5000) DSPE, and folate-lipid (without PEG linker) in microemulsions. In vitro studies were done in a human nasopharyngeal cell line, KB, which overexpresses the folate receptor (FR), and a human hepatoblastoma cell line, [FR(-)] HepG2. In vivo experiments were done in a KB xenograft by systemic administration of folate linked microemulsions loading ACM. RESULTS: The association of folate-linked microemulsions to KB cells could be blocked by 2 mmol/L free folic acid. Selective FR-mediated cytotoxicity of folate-linked microemulsions loading ACM was obtained in KB but not in HepG2 cells. The association of the folate PEG(5000)-linked microemulsion and folate-PEG(2000)-linked microemulsion with the cells was 200- and 4-fold higher, whereas their cytotoxicity was 90- and 3.5-fold higher than those of nonfolate microemulsion, respectively. The folate-PEG(5000) linked microemulsions showed 2.6-fold higher accumulation in solid tumors 24 hours after i.v. injection and greater tumor growth inhibition than free ACM. CONCLUSION: These findings suggest that a folate-linked microemulsion is feasible for tumor-targeted ACM delivery. This study shows that folate modification with a sufficiently long PEG chain on emulsions is an effective way of targeting emulsion to tumor cells. PMID- 15756029 TI - Farnesyltransferase inhibitors and human malignant pleural mesothelioma: a first step comparative translational study. AB - It is known that the potential clinical use of farnesyltransferase inhibitors (FTI) could be expanded to include cancers harboring activated receptor tyrosine kinases. Approximately 70% of malignant pleural mesotheliomas (MPM) overexpress epidermal growth factor receptors (EGFR) and a subset express both EGFR and transforming growth factor alpha (TGF-alpha), suggesting an autocrine role for EGFR in MPM. We checked on MPM cells (10 human cell lines, 11 primary cultures obtained by human biopsies, and 7 short-term normal mesothelial cell cultures) concerning the following: (a) the relative overexpression of EGFR (Western blotting, flow cytometry, immunohistochemistry), (b) the relative expression of EGFR ligands (EGF, amphiregulin, TGF-alpha, ELISA), (c) the relative increase of the activated form of Ras (Ras-bound GTP) after EGF stimulation (Ras activation assay), (d) the efficacy of five different FTIs (HDJ2 prenylation, cell cytotoxicity, and apoptosis using ApopTag and gel ladder). EGFR was overexpressed in MPM cells compared with normal pleural mesothelial cells in equivalent levels as in non-small cell lung cancer cells A459. MPM cells constitutively expressed EGFR ligands; however, Ras activation was attenuated at high EGF concentrations (100 ng/mL). Growth of MPM cells was substantially not affected by treatment with different FTIs (SCH66336, BMS-214662, R115777, RPR-115135, and Manumycin). Among these, BMS-214662 was the only one moderately active. BMS-214662 triggered apoptosis in a small fraction of cells (not higher than 30%) that was paralleled by a slight decrease in the levels of TGF-alpha secreted by treated MPM cells. Our data highlighted the concept that the same signaling pathway can be regulated in different ways and these regulations can differ between different cells of different origin. PMID- 15756030 TI - Flavopiridol blocks integrin-mediated survival in dormant breast cancer cells. AB - PURPOSE: Breast cancer micrometastases in the bone marrow are resistant to chemotherapy. They can remain dormant for years before some begin to proliferate. We seek to understand survival mechanisms and develop targeted approaches to eliminating these cells. EXPERIMENTAL DESIGN: In an in vitro model of dormancy, basic fibroblast growth factor 2 (FGF-2), abundant in the bone marrow, inhibits the growth of well-differentiated cells in the 2- to 10-cell stage and up regulates integrin alpha(5)beta(1). Through this integrin, cells bind fibronectin, spread out, and acquire a survival advantage, partly through activation of the phosphatidylinositol 3-kinase/Akt pathway. We investigated the effects of Taxotere, flavopiridol, and mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase and p38 inhibitors on survival of dormant clones and that of flavopiridol on expression of integrins, adhesion strength, and phosphorylation of Akt, ERK 1/2, and p38. RESULTS: Dormant MCF-7 and T-47D cell clones were resistant to Taxotere concentrations 10-fold higher than needed to eliminate growing clones but were almost completely eradicated by 200 nmol/L flavopiridol. Flavopiridol caused a decrease in FGF-2-induced expression of integrins, including alpha(5) and beta(1), and decreased FGF-2-induced specific adhesion to fibronectin. It diminished Akt phosphorylation, but reexpression of active Akt was not sufficient to reverse dormant clone inhibition. Flavopiridol did not affect phosphorylation of ERK 1/2 and p38 but diminished total protein levels. Chemical inhibition of these pathways partially abrogated dormant clone survival. CONCLUSIONS: Flavopiridol has pleiotropic effects on key targets involved with survival of dormant breast cancer cells and may represent a useful approach to eliminating cells dependent on multiple signal pathways for survival. PMID- 15756031 TI - Antisense inhibition of methylenetetrahydrofolate reductase reduces cancer cell survival in vitro and tumor growth in vivo. AB - PURPOSE: Many cancer lines are methionine dependent and decrease proliferation when methionine supply is limited. Methylenetetrahydrofolate reductase (MTHFR) generates the folate derivative for homocysteine remethylation to methionine. We investigated the effect of antisense-mediated inhibition of MTHFR on survival of human cancer cells. EXPERIMENTAL DESIGN: We examined the in vitro and in vivo anticancer effects of a combination of MTHFR antisense and standard cytotoxic drugs. RESULTS: Specific antisense against MTHFR (EX5) showed significant inhibitory effects on growth of human colon, lung, breast, prostate, and neuroblastoma tumor cells in vitro compared with that of the control oligonucleotide. Cytotoxic drugs (5-fluorouracil, cisplatin, or paclitaxel) potentiated the effect of EX5. In vivo, antisense alone or in combination with cytotoxic drugs inhibited the growth of human colon and lung carcinoma xenografts. In comparison with control oligonucleotide, treatment with EX5 inhibited growth of colon tumors and lung tumors by 60% and 45%, respectively. EX5 with 5-fluorouracil decreased growth of colon tumors by an additional 30% compared with EX5 alone, and EX5 with cisplatin decreased growth of lung tumors by an additional 40% compared with cisplatin alone. Growth inhibition by EX5 was associated with decreased amounts of MTHFR protein and with increased amounts of an apoptosis marker. CONCLUSIONS: Our results confirm that MTHFR inhibition decreases tumor growth and suggest that inhibition of MTHFR by antisense or small molecules may be a novel anticancer approach. PMID- 15756032 TI - Gene expression profiling of the irinotecan pathway in colorectal cancer. AB - The exact mechanism responsible for large variation of response to chemotherapy remains unclear. This study profiled the gene expression for the entire irinotecan pathway to provide insights into individualized cancer therapy. The RNA expressions of 24 irinotecan pathway genes were measured in paired tumor and normal tissues from 52 patients with Dukes' C colorectal cancer using a real-time quantitative reverse transcription-PCR assay. The relative expression levels across the 24 pathway genes varied considerably, with a 441-fold range from highest to lowest expression levels for the tumor tissues and a 934-fold range for the normal tissues. Interpatient variability was also quite large, with a 33.6 median fold change in the tumor tissue genes and a 30.1 median fold change in the normal tissue genes. Six of the 24 irinotecan pathway genes had dramatically lower expression levels in the tumor samples than did the genes in the normal tissues (median range, 1.28-4.39 folds; P = 0.001-0.029). Eight genes had significantly higher levels (median range, 1.35-2.42 folds; P = 0.001-0.011). Using hierarchical clustering, three gene clusters and three patient groups were observed with high similarity indices by the RNA expressions in colorectal tumors. The three patient groups had no unique clinical pathologic features but could be differentiated by the statistically significant differences in RNA expression level of seven genes. Our study indicates that gene expression profiling could be valuable for predicting tumor response to chemotherapy and for tailoring therapy to individual cancer patients. PMID- 15756033 TI - Combination therapy enhances the inhibition of tumor growth with the fully human anti-type 1 insulin-like growth factor receptor monoclonal antibody CP-751,871. AB - PURPOSE: The insulin-like growth factor (IGF) signaling pathway is implicated in cellular mitogenesis, angiogenesis, tumor cell survival, and tumorigenesis. Inhibition of this pathway results in decreased cell growth, inhibition of tumor formation in animal models, and increased apoptosis in cells treated with cytotoxic chemotherapy. We generated and characterized a human monoclonal antibody that targeted the IGF receptor. EXPERIMENTAL DESIGN: By use of XenoMouse technology, we generated CP-751,871, a fully human IgG2 antibody with high affinity (K(d) = 1.5 nmol/L) for human IGF-1R and evaluated its biological, pharmacologic, and antitumor properties. RESULTS: This antibody blocks binding of IGF-1 to its receptor (IC(50) 1.8 nmol/L), IGF-1-induced receptor autophosphorylation (IC(50) 0.42 nmol/L) and induced the down-regulation of IGF 1R in vitro and in tumor xenografts. The extent of IGF-1R down-regulation in vivo was proportional to CP-751,871 concentrations in the serum of tumor-bearing mice. Pharmacokinetic profiles in cynomolgus monkeys indicated a close to linear increase of exposure following i.v. dosing of antibody in the range of 3 to 100 mg/kg. CP-751,871 showed significant antitumor activity both as a single agent and in combination with Adriamycin, 5-fluorouracil, or tamoxifen in multiple tumor models. A biomarker assay was developed to establish the relationship between circulating antibody concentrations and down-regulation of IGF-1R in peripheral blood cells. The concentration of CP-751,871 required to down-regulate 50% of IGF-1R on peripheral blood cells was 0.3 nmol/L. CONCLUSION: These data suggest that inhibition of the IGF cascade by use of this monoclonal antibody may be of clinical benefit in the treatment of human cancers. PMID- 15756034 TI - Telomere shortening and telomerase reverse transcriptase expression in preinvasive bronchial lesions. AB - PURPOSE: Telomerase, a ribonucleoprotein complex whose activity is related to the expression of its catalytic subunit human telomerase reverse transcriptase (hTERT), restores telomere length in tumor cells and enables immortality after p53/Rb inactivation has been achieved. To determine the timing of hTERT derepression during bronchial carcinogenesis and its relationship with telomere shortening and the p53/Rb pathway alterations, we did an immunohistochemical and in situ hybridization study in preinvasive and invasive bronchial lesions. EXPERIMENTAL DESIGN: hTERT, P53, P16, cyclin D1, Bax-to-Bcl2 ratio, and Ki67 immunostainings were done in 106 preneoplastic lesions and in paired lung carcinoma and normal bronchial mucosae. Concomitantly, hTERT mRNA levels and qualitative telomere shortening were assessed by in situ hybridization and fluorescence in situ hybridization, respectively, in a subset of preneoplastic and neoplastic lesions. RESULTS: Telomerase was increasingly expressed from normal epithelium to squamous metaplasia, dysplasia, and carcinoma in situ, and decreased in invasive carcinoma (P < 0.0001), with a direct correlation between protein and mRNA levels of expression (P < 0.0001). hTERT expression was directly correlated with P53, Ki67, and Bcl2-to-Bax ratio, suggesting a coupling between telomerase reactivation, proliferation, and resistance to apoptosis. Telomere signals significantly decreased as early as squamous metaplasia and progressively increased over the spectrum of preneoplastic lesions. CONCLUSIONS: Telomere shortening represents an early genetic abnormality in bronchial carcinogenesis, preceding telomerase expression and p53/Rb inactivation, which predominate in high-grade preinvasive lesions. PMID- 15756035 TI - Effect of the synthetic retinoid fenretinide on circulating free prostate specific antigen, insulin-like growth factor-I, and insulin-like growth factor binding protein-3 levels in men with superficial bladder cancer. AB - PURPOSE: Fenretinide (4-HPR) is a synthetic retinoid that has shown a preventive activity in prostate cancer animal models. EXPERIMENTAL DESIGN: We measured the changes in total and free prostate-specific antigen (PSA) and its association with insulin-like growth factor I (IGF-I) and IGFBP-3 levels after 1 year of treatment in 24 subjects given 4-HPR and 24 control subjects enrolled in a randomized bladder cancer prevention trial. RESULTS: No significant effect of 4 HPR was observed on total and free fraction of PSA levels. The median percentage [95 confidence interval (95% CI)] change for % free PSA and total PSA in the 4 HPR and the control group were, respectively, 7.6 (95% CI, -4.0 to 69.3) versus 5.1 (95% CI, -21.4 to 59.8) and -7.8 (95% CI, -18.2 to 52.5) versus -12.3 (95% CI, -44.6 to 9.6). However, in patients ages <60 years, there was a trend to an increase of total free PSA and % free PSA after treatment with 4-HPR that was different from a trend to a decrease in the control group (P = 0.002 and 0.052, respectively). The interaction between age and treatment was statistically significant on free PSA (P = 0.001). A similar pattern was noted with smoking status (P = 0.011 for the interaction on free PSA). No association was observed between PSA levels and IGF-I or IGFBP-3 levels. CONCLUSIONS: We conclude that 4 HPR has no significant effect on circulating PSA, but it increases significantly free PSA levels in subjects younger than 60 years and in nonsmokers. These effects might support an activity in prostate cancer prevention but further studies are required. PMID- 15756037 TI - Measles control: current trends & recommendations. PMID- 15756036 TI - Overexpression of 5-lipoxygenase and cyclooxygenase 2 in hamster and human oral cancer and chemopreventive effects of zileuton and celecoxib. AB - PURPOSE: Previous studies have suggested an important role of aberrant arachidonic acid metabolism, especially the cyclooxygenase (Cox) pathway, in oral carcinogenesis. However, it is unknown whether the 5-lipoxygenase (5-Lox) pathway contributes to oral carcinogenesis, and whether combination of inhibitors of both pathways may have synergistic or additive effects of chemoprevention. EXPERIMENTAL DESIGN: 5-Lox expression was examined in 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster and human oral cancer tissues by immunohistochemistry, and Cox2 expression was investigated in hamster oral tissues using in situ hybridization. Zileuton (a specific 5-Lox inhibitor) and celecoxib (a specific Cox2 inhibitor), either alone or in combination, were investigated for their chemopreventive effects on the DMBA-induced hamster model at the post-initiation stage through topical application. RESULTS: 5-Lox was overexpressed during oral carcinogenesis in hamsters and humans, as well as Cox2 in the hamster tissues. In a chemoprevention study using the post-initiation DMBA model, incidence of hamster oral squamous cell carcinoma was reduced from 76.9% (20 of 26) to 45.8% (11 of 24, P < 0.05) and 32.1% (9 of 28, P < 0.01) by 3% and 6% topical zileuton, respectively; and to 57.6% (15 of 26, P > 0.05) and 50% (12 of 24, P < 0.05) by 3% and 6% topical celecoxib, respectively. When used in combination, celecoxib and zileuton (3% of each) had an additive inhibitory effect on the incidence of squamous cell carcinoma (36%, 9 of 25, P < 0.01). Other pathologic variables and the levels of leukotriene B4 and prostaglandin E2 of the hamster tissues were reduced as well. CONCLUSIONS: The results clearly showed that both 5-Lox and Cox2 played important roles in oral carcinogenesis. Zileuton and celecoxib prevented oral carcinogenesis at the post-initiation stage through their inhibitory effects on arachidonic acid metabolism. PMID- 15756038 TI - Platelet endothelial cell adhesion molecule-1 (PECAM-1) & coronary heart disease. PMID- 15756039 TI - Can we reduce preterm births? PMID- 15756040 TI - The challenge of antibiotic resistance: need to contemplate. AB - "Survival of the fittest " holds good for men and animals as also for bacteria. A majority of bacteria in nature are nonpathogenic, a large number of them, live as commensals on our body leading a symbiotic existence. A limited population of bacteria which has became pathogenic was also sensitive to antibiotics to begin with. It is the man made antibiotic pressure, which has led to the emergence and spread of resistant genes amongst bacteria. Despite the availability of a large arsenal of antibiotics, the ability of bacteria to become resistant to antibacterial agents is amazing. This is more evident in the hospital settings where the antibiotic usage is maximum. The use of antibiotics is widespread in clinical medicine, agriculture, aquaculture, veterinary practice, poultry and even in household products. The major reason for this is the inappropriate use of antibiotics due to a lack of uniform policy and disregard to hospital infection control practices. The antibiotic cover provided by newer antibiotics has been an important factor responsible for the emergence of multi-drug resistant bacteria. Bacterial infections increase the morbidity and mortality, increase the cost of treatment, and prolong hospital stay adding to the economical burden on the nation. The problem is further compounded by the lack of education and " over the counter " availability of antibiotics in developing countries. Antibiotic resistance is now all pervasive with the developed world as much vulnerable to the problem. Despite advancement in medical technology for diagnosis and patient care, a person can still die of an infection caused by a multi-drug resistant bacteria. It is time to think, plan and formulate a strong antibiotic policy to address the burgeoning hospital infection. PMID- 15756042 TI - Is dengue emerging as a major public health problem? AB - BACKGROUND & OBJECTIVES: Diagnosis of dengue infection is easily and best accomplished by demonstration of specific IgM antibodies in blood. We analyzed retrospectively the dengue IgM seropositivity available for samples obtained over a period of five year (1999-2003) from patients with suspected dengue fever (DF) like illness to investigate whether there was an overall increase in the dengue IgM prevalence over this period. METHODS: Serum samples from a total of 1426 individuals (suspected dengue cases) obtained over five year were tested for dengue specific IgM antibodies. Of the 1426 patients, 693 were adults (>15 yr) and 694 children (<15 yr) (excluding 39 individuals whose age was not known). There were 807 males and 610 females (excluding 9 individuals whose status on sex was unknown). RESULTS: A total of 423 (29.7%) samples were positive for dengue IgM over the five year period. Overall, there was a significant increase in the percentage of dengue IgM positive individuals over the this period (P<0.001). When the individuals were grouped into children (<15 yr) and adults (>15 yr), a significant increase in the number of dengue IgM positive individuals was noticed only in children (P<0.001) and not in adults. When the individuals were grouped into males and females, a significant increase in the number of dengue IgM positive individuals was noticed in both the sexes (P<0.03). Month-wise analysis of the dengue IgM positivity rates indicated the year-wide occurrence of dengue. A total of 158 (41%) of the dengue IgM positive individuals showed positivity for dengue IgG also suggestive of a secondary heterotypic infection. INTERPRETATION & CONCLUSION: The overall significant increase in dengue IgM seropositivity among the suspected cases indicates an increase in dengue virus activity, raising the question whether dengue is emerging/re-emerging as a major health problem in southern India. Increase in probable secondary infection (as evidenced by dual positivity for dengue IgM and IgG) seen in this study is also a point of concern. Such an increase especially in a country like ours where multiple serotypes are prevalent, raises concern over probable increase in the incidences of the more serious DHF/DSS. As this report could well be an underestimate of true incidence, the alarming increase observed in 2003, may be a warning/indication of epidemics to come soon that merits serious consideration. PMID- 15756041 TI - Association of Leu125Val polymorphism of platelet endothelial cell adhesion molecule-1 (PECAM-1) gene & soluble level of PECAM-1 with coronary artery disease in Asian Indians. AB - BACKGROUND & OBJECTIVES: Platelet endothelial cell adhesion molecule-1 (PECAM-1) plays a key role in the transendothelial migration of circulating leukocytes (diapedesis) during vascular inflammation. We hypothesized that genetic variation and the level of soluble PECAM-1 could be associated with the development of atherosclerosis and conducted a study on gene polymorphisms of PECAM-1 and soluble PECAM-1 levels in Asian Indian patients with coronary artery disease (CAD) in Singapore. METHODS: Of the 137 angiographically confirmed patients (> or =70% stenosis) of CAD and 110 controls in Asian Indian population, two single nucleotide polymorphisms (SNPs) of PECAM-1 gene, C+373G (Leu125Val) at exon 3 and G+1688A (Ser563Asn) at exon 8 were analyzed by polymerase chain reaction restriction fragment-length polymorphism (PCR-RFLP) strategy. In addition, plasma soluble PECAM-1, P-selection and lipid profile were measured. Chi- square test and student t test were adopted to compare categorical and continuous variables, respectively. RESULTS: A significant decrease in C allele frequency but increase in G allele frequency of the Leu125Val (C/G) polymorphism were observed in CAD patients as compared with controls (0.54/0.46 vs 0.663/0.337 respectively, P=0.008). Alteration in genotype distributions (CC, CG and GG) of the Leu125Val polymorphism between CAD patients and controls (P=0.009) was also significant. A similar trend was observed on the allele frequencies (G/A) and genotype distributions of Ser563Asn (G/A) polymorphism, though the difference did not reach significance. On the other hand, plasma level of soluble PECAM-1 (sPECAM-1) was markedly elevated in CAD patients (P=0.006), and associated with soluble P selectin and lipid profiles. INTERPRETATION & CONCLUSION: Our study showed that Leu125Val polymorphism of PECAM-1 gene and elevated soluble PECAM-1 were related to severe coronary artery stenosis in CAD patients of Asian Indian origin in Singapore. Our data also suggest that PECAM-1 plays an important role in the development of atherosclerosis. PMID- 15756043 TI - Modified cold Z-N staining for presumptive identification of Brucella. AB - Brucellosis is one of the most common zoonotic diseases in India. Growth of Brucella is slow and needs meticulous biochemical tests and antisera for confirmation of the isolates. In the present study modified cold ZN staining was tried on the broth cultures for early presumptive identification of Brucella growth. Blood cultures were attempted in 22 seropositive patients. In the five blood cultures positive for Brucella, acid-fast coccobacilli were seen in broth smears stained with modified cold ZN stain, thus providing presumptive identification of Brucella growth. Acid-fast bacteria were not seen in the broth smears of the remaining 17 broths negative for Brucella growth. The method is simple, reliable and reproducible and needs to be evaluated further on a larger sample. PMID- 15756044 TI - Erythrocyte antioxidant enzymes in Parkinson's disease. AB - BACKGROUND & OBJECTIVES: Oxidative stress is incriminated to play a central role in the pathogenesis of Parkinson's disease (PD). Oxidative stress, to which neurons are highly susceptible, is also known to induce oxidative changes in human red blood cells (RBCs), in vivo and in vitro. Earlier studies on oxidative stress in RBCs in patients with PD have yielded controversial results claiming unaltered activity to reduced activity. Using RBC as a model, we have undertaken this study to ascertain the possibility of oxidative damage to the RBCs in PD by measuring the cytosolic antioxidant enzymes viz., superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (G-Px) and glucose-6-phosphate dehydrogenase (G6PD). METHODS: Activities of antioxidant enzymes were measured in erythrocytes of 115 PD patients and 37 normal age-matched healthy persons as controls. Enzymes activities were correlated with age of patients, age of onset of disease, duration of disease, United Parkinson's Disease Rating Scale (UPDRS) and Hoehn and Yahr stage. RESULTS: The SOD, CAT, G-Px and G6PD activities were significantly lower in patients with PD compared to the control. A significant (P<0.05) negative correlation of enzyme activities with Hoehn and Yahr stage of the disease and also with UPDRS score was found. INTERPRETATION & CONCLUSION: Results of the present study showed involvement of oxidative stress as one of the risk factors, which can initiate and/or promote neurodegeneration in PD and was correlated to the severity of the disease. PMID- 15756045 TI - Toxicity of a mosquitocidal metabolite of Pseudomonas fluorescens on larvae & pupae of the house fly, Musca domestica. AB - BACKGROUND & OBJECTIVES: Biological control through the use of parasitoids and pathogens is one of the alternatives to the use of chemical pesticides for control of insects of public health importance. At the Vector Control Research Centre, a liquid formulation developed using the metabolite of a Pseudomonas fluorescens strain was found to be lethal to larvae as well as pupae of vector mosquitoes. The lethal fraction of the metabolite is a protein with a molecular mass of 44 kDa and toxicity studies showed that it is safe to mammals. In the present study, this formulation was evaluated against immatures of the common house fly, Musca domestica, to find out whether it could be developed into a potential biocontrol tool. METHODS: Early second instar larvae of house fly were introduced into rearing medium incorporated with the formulation at concentrations of 1, 5, 10, 15, 20 and 25 per cent, which were equivalent to respectively 1.13, 5.63, 11.25, 16.88, 22.50 and 28.13 microg of the toxic protein/ g of rearing medium. Mortality was monitored until the emergence of adult house fly. Net mortality of larvae and pupae were calculated and the LC50 and LC90 values were determined through probit regression analysis. RESULTS: Larval mortality was obtained from day 3 to 6 post-treatment. Net mortality of larvae was higher at the concentration of 20 than at 25 per cent. However, it was higher at 25 per cent on day 5 and continued to day 6 when there was no larval mortality at other concentrations. The net mortality of pupae was higher than that of larvae at all the concentrations except at 20 per cent. The LC50 and LC90 values calculated from the net mortality of larvae and pupae together, from day 1 to 12 post-treatment, were respectively, 8.25 and 51.79 microg protein/g of the fly rearing medium. INTERPRETATION & CONCLUSION: The formulation prepared from the exotoxin of P. fluorescens was toxic to the house fly. Pupae were more susceptible than larvae and the activity of the toxin might have been through cuticular absorption. The results are indicative of the possibility of development of the mosquitocidal metabolite for house fly control through appropriate field evaluations. PMID- 15756046 TI - Ritodrine in the treatment of preterm labour: a meta-analysis. AB - BACKGROUND & OBJECTIVES: Results on the effect of ritodrine in the treatment of preterm labour are different all over the world. Therefore, a concrete conclusion cannot be drawn from a single randomized control trial (RCT). In this study, we selected a large number of RCTs worldwide on the treatment of preterm labour comparing ritodrine and placebo or magnesium sulfate and by applying meta analysis, evaluated the effect of ritodrine in the treatment of preterm labour. METHODS: We searched and identified 20 RCTs about ritodrine versus placebo, and ritodrine versus magnesium sulphate in the main medical data resources (MEDLINE, PubMed, CBMdisc, Cochrane Library and EMBASE) from January 1970 to December 2001 published in English and Chinese literature. We abstracted data about delay of gestation, gestational age at delivery, birth weight, severe neonatal respiratory system diseases, perinatal deaths, and administration to contraction-ceased interval, etc. RESULTS: The odds ratio (OR) of 1 day (d) delay in 10 RCTs on ritodrine in preterm labour versus placebo was 2.95 (95%CI 2.15, 4.04), and OR of 2d delay was 1.91 (95%CI 1.49, 2.45), both differences were statistically significant. There were no significant difference on the OR of birth > or =37 wk, birth weight > or =2500 g, severe respiratory morbidity and perinatal mortality. The OR of 2d delay of four RCTs from abroad on ritodrine versus magnesium sulphate was 1.56 (95%CI 0.62, 3.94), and OR of 7d delay was 1.45 (95%CI 0.80, 2.62), both without significant difference. But the combined estimated rate difference (CERD) of side effects interruption rate was 12 per cent with significant difference. There were six Chinese RCTs on ritodrine and magnesium sulphate. The OR of 1d delay was 3.95 (95%CI 1.98, 7.88), and OR of neonatal death was 0.45 (95%CI 0.22, 0.93), both with significant difference. The average administration to contractionceased interval were 2.5 and 6.3 h, respectively, with CERD being -4.2 h. INTERPRETATION & CONCLUSION: In treating preterm labour, ritodrine can significantly prolong a short interval more quickly but with relatively more side effects than magnesium sulphate. Stratified RCTs for different gestational ages and different labour stages should be designed for further study. PMID- 15756047 TI - Resurgence of Vibrio cholerae O139 in Rohtak. PMID- 15756049 TI - Glucocorticoid-induced osteoporosis: from basic mechanisms to clinical aspects. AB - Glucocorticoid (GC)-induced osteoporosis (GCOP) is the most common cause of osteoporosis in adults aged 20-45 years as well as the most common cause of iatrogenic osteoporosis. GC excess, either endogenous or exogenous, induces bone loss in 30-50% of cases. Indeed, bone loss leading to fractures is perhaps the most incapacitating, sometimes partially irreversible, complication of GC therapy. Nevertheless, GCOP is often underdiagnosed and left untreated. The following article provides an update on the cellular and molecular mechanisms implicated in the pathophysiology of GC-induced bone loss, as well as some guidelines on diagnostic, preventive and therapeutic strategies for this medical condition, in an effort to promote a better knowledge and greater awareness of GCOP by both the patient and the physician. PMID- 15756050 TI - Induction of experimental allergic encephalomyelitis in a low-susceptible Albino Oxford rat strain by somatostatin analogue SMS 201-995. AB - OBJECTIVE: The effect of the somatostatin analogue SMS 201-995 (octreotide; OCT) on the course of experimental allergic encephalomyelitis (EAE) in the relatively resistant Albino Oxford (AO) strain of rats was studied. METHODS: Animals were actively immunized with bovine brain homogenate in complete Freund's adjuvant. OCT was given subcutaneously in the hind legs on days 7, 8 and 9 after immunization, at a dose of 3 x 5 microg/kg/day. Rats in control groups were treated with saline or were left untreated. EAE was scored clinically and immunophenotypically, estimating by flow cytometry the changes in the popliteal lymph nodes (PLN) and spleen and monitoring immunohistologically the brain sections of rats recovered from disease. RESULTS: In control AO rats, EAE was induced in only 2 of 22 rats (9%). In OCT-treated rats, however, EAE developed in 11 of 20 rats (55%), in comparison with 3 of 17 saline-treated animals (17%) (p <0.05). In PLN of OCT-treated rats during the clinical course of EAE, a decreased proportion of OX8+ cells was seen, followed by increases in OX39+ and W3/25+ cells on days 17 and 26. In spleen, OCT decreased the proportion of OX1+, OX39+ and OX8+ cells (on days 12 and/or 17), and increased the proportion of OX39+ cells on days 26 and 31. In the brain sections of saline-treated rats recovered from EAE, numerous Mac-1+, Mac-3+ and OX8+ cells were found. These cells were, however, absent in OCT-treated rats; instead, several W3/25+ cells were noticed. CONCLUSIONS: These data imply that OCT increases the susceptibility of AO rats to EAE, interfering with specific and/or nonspecific defense mechanisms operating in both the initial and recovery phase of EAE. PMID- 15756051 TI - Immunological study of IFNbeta-1a-treated and untreated multiple sclerosis patients: clarifying IFNbeta mechanisms and establishing specific dendritic cell immunotherapy. AB - OBJECTIVES: A comparative immunological evaluation of multiple sclerosis (MS) patients receiving IFNbeta treatment and patients who are not receiving treatment may help clarify IFNbeta neurological mechanisms and lead the way to an effective dendritic cell (DC) immunotherapy. This type of study helps clarify the pathological function of T cells and DCs within the TH1/TH2/TH3 network as well as the specific interactions between TH1/TH2/TH3 cytokines implicated in MS pathological mechanisms and determine the best way of reestablishing the TH1/TH2/TH3 network equilibrium. METHODS: We studied network interactions between TH1/TH2/TH3 cytokine levels in serum and supernatants of whole blood and CD14+ monocyte-derived DCs in the remission phase of the disease and in correlation to the Expanded Disability Status Scale (EDSS). RESULTS: We found that TH1 dysregulation results in a disruption of the maturation and activation of dendritic and T cells, and a lack of T-regulating cells for the induction of self tolerance; IFNbeta mechanisms restore regulation by reestablishing the network balance but fail to resolve the disease completely due to in vivo IL12p70 network interactions leading to the deletion of self-aggressive cells. CONCLUSIONS: Our results indicate that a specific DC immunotherapy could cure rather than treat MS. The best point to reestablish the normal physiological cycle is at the immature DC stage which can be done in vitro with treated peripheral blood CD14+ cells and used in vivo to stimulate the expansion of specific regulatory T cells. PMID- 15756052 TI - Dopamine exerts no acute effects on Kv1.3 in activated encephalitogenic T cells. AB - Apart from a central function in the extrapyramidal motor system, dopamine has been suggested to play a role in neuroimmune interactions. Particularly in diseases of the central nervous system, such as multiple sclerosis, alterations in dopamine homeostasis might have immunological consequences. We investigated potential effects of dopamine stabilized by ascorbic acid on specifically activated encephalitogenic T cells at the peak of activation. Those cells exhibited an upregulation of voltage-sensitive K+ channels which play a role in many neurotransmitter responses of lymphocytes and fulfilled a prerequisite to respond to dopamine, i.e. stable expression of mRNA for dopamine receptors DRD1, DRD2 and DRD3. However, whole-cell and perforated whole-cell recordings revealed no change in voltage-sensitive K+ currents. Moreover, T cell proliferation was not changed in the presence of dopamine. Previously reported dopamine effects on T cells may be explained by a comparatively lower activation of the cells under investigation, suggesting an activation dependence of dopamine effects that may not be mediated by K+ channels. Alternatively, the occurrence of dopamine degradation products under unprotected conditions may account for the changes reported. Nevertheless, care should be taken when using the dopamine-protecting anti-oxidant ascorbic acid, since we found that it markedly inhibited both K+ currents and lymphocyte proliferation at higher concentrations. PMID- 15756053 TI - ICAM G241A polymorphism and soluble ICAM-1 serum levels: evidence for an active immune process in schizophrenia. AB - OBJECTIVES: We have previously reported reduced serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in schizophrenic patients. A single nucleotide polymorphism (SNP) of the ICAM-1 gene was described at position 241. The G-->A SNP results in a nonsynonymous amino acid exchange of the ICAM-1 protein, and the A allele was shown to be also associated with several immunological disorders like rheumatoid arthritis. METHODS: We investigated 70 schizophrenic patients and 128 unrelated healthy control persons regarding the relationship between the serum levels of sICAM-1 and the ICAM-1 G214A polymorphism. RESULTS: We were able to replicate our previous finding of reduced sICAM-1 levels in schizophrenia. Healthy control persons carrying the polymorphic A allele showed markedly lower sICAM-1 serum levels than carriers of the homozygous GG wild type (p < 0.004). In contrast, no significant difference in the sICAM-1 serum levels were seen regarding the G241A genotype distribution in schizophrenic patients. CONCLUSION: We hypothesize that the biochemical effect of the G241A SNP is masked in schizophrenic patients, indicating a disease-related mechanism leading to reduced levels of sICAM-1 in schizophrenia. PMID- 15756054 TI - Postoperative interleukin-6 and cortisol concentrations in elderly patients with postoperative confusion. AB - OBJECTIVE: To investigate changes in plasma interleukin (IL)-6, cortisol or noradrenaline concentrations after surgery in elderly patients with postoperative confusion. METHODS: We studied 80 patients aged 70-90 years undergoing abdominal surgery and measured plasma IL-6, cortisol or noradrenaline concentrations before surgery, at the end of surgery, and 24 and 48 h after surgery. RESULTS: Plasma IL 6 concentrations in elderly patients with postoperative confusion were 83.2 +/- 30.5, 49.3 +/- 14.1 and 42.9 +/- 19.4 pg.ml(-1) at the end of surgery, and 24 and 48 h after surgery, respectively, being significantly higher than in elderly patients without postoperative confusion (58.0 +/- 37.5, 36.1 +/- 20.0 and 28.2 +/- 16.7 pg.ml(-1)). Plasma cortisol concentrations in elderly patients with postoperative confusion (42.2 +/- 7.8, 38.3 +/- 8.3 and 33.1 +/- 8.4 microg.dl( 1) at the end of surgery, and 24 and 48 h after surgery, respectively) were significantly higher than in elderly patients without postoperative confusion (32.9 +/- 6.7, 30.4 +/- 8.6 and 25.6 +/- 6.5 microg.dl(-1), respectively). There were no significant differences in plasma norepinephrine concentrations at all sampling points between elderly patients with and without postoperative confusion. However, plasma IL-6 and cortisol concentrations were related in elderly patients with postoperative confusion (at the end of surgery and 24 and 48 h after surgery). However, there was no relationship between plasma IL-6 and cortisol concentrations 24 and 48 h after surgery in elderly patients without postoperative confusion. CONCLUSION: Elderly patients with postoperative confusion had increased plasma IL-6 and cortisol concentrations. The interaction between IL-6 and cortisol after surgery is associated with developing postoperative confusion in the elderly patients. PMID- 15756055 TI - Effect of the macrolide antibacterial drug, tylosin, on TNBS-induced colitis in the rat. AB - Bacterial antigens, such as intestinal microflora, are known to play a role in the pathogenesis of human inflammatory bowel disease (IBD). Tylosin, a macrolide antimicrobial agent, has proven to be effective in cat and dog chronic colitis, but the reasons underlying this efficacy are still unclear. In the present study we evaluated the effects of tylosin on 2,4,6-trinitrobenzenesulfonic acid (TNBS) induced colitis in the rat, in comparison with the antibacterial drug metronidazole and the corticosteroid budesonide. Colitis was induced by a single intrarectal administration of 10 mg TNBS under light ether anesthesia. Tylosin (20 mg/kg twice a day), metronidazole (160 mg/kg twice a day) and budesonide (500 microg/kg once a day) were given orally for up to 6 days to separate groups of rats. The animals were sacrificed after 6 days and colonic lesions evaluated (colon weight, macroscopic and histologic damage, myeloperoxidase activity). Tylosin and metronidazole significantly lowered macroscopic lesion score, reduced colon weight, the severity of histologic lesions and myeloperoxidase activity; budesonide did not significantly change the parameters of colonic inflammation. These data indicate a protective effect of tylosin against intestinal inflammation, suggesting a major role for bacteria, anaerobes in particular, in the development of TNBS-induced mucosal damage. PMID- 15756056 TI - Nucleosomes in pancreatic cancer patients during radiochemotherapy. AB - Nucleosomes appear spontaneously in elevated concentrations in the serum of patients with malignant diseases as well as during chemo- and radiotherapy. We analyzed whether their kinetics show typical characteristics during radiochemotherapy and enable an early estimation of therapy efficacy. We used the Cell Death Detection Elisa plus (Roche Diagnostics) and investigated the course of nucleosomes in the serum of 32 patients with a local stage of pancreatic cancer who were treated with radiochemotherapy for several weeks. Ten of them received postsurgical therapy, 21 received primary therapy and 1 received therapy for local relapse. Blood was taken before the beginning of therapy, daily during the first week, once weekly during the following weeks and at the end of radiochemotherapy. The response to therapy was defined according to the kinetics of CA 19-9: a decrease of CA 19-9 > or =50% after radiochemotherapy was considered as 'remission'; an increase of > or =100% (which was confirmed by two following values) was defined as 'progression'. Patients with 'stable disease' ranged intermediately. Most of the examined patients showed a decrease of the concentration of nucleosomes within 6 h after the first dose of radiation. Afterwards, nucleosome levels increased rapidly, reaching their maximum during the following days. Patients receiving postsurgery, primary or relapse therapies did not show significant differences in nucleosome values during the time of treatment. Single nucleosome values, measured at 6, 24 and 48 h after the application of therapy, could not discriminate significantly between patients with no progression and those with progression of disease. However, the area under the curve of the first 3 days, which integrated all variables of the initial therapeutic phase, showed a significant correlation with the progression free interval (p=0.008). Our results indicate that the area under the curve of nucleosomes during the initial phase of radiochemotherapy could be valuable for the early prediction of the progression-free interval. PMID- 15756057 TI - Immunohistochemical expression of Bcl-2, Ki-67, and p21 in patients with papillary thyroid cancer. AB - Papillary thyroid cancer (PTC) is a slow-growing tumor with a favorable outcome. Still, some low-risk patients develop local or distant metastases and eventually die from their disease. Many molecular markers are involved in proliferation and apoptosis, including Bcl-2, Ki-67, and p21. Because age over 45 is the most important determinant of a poor survival, we analyzed whether the expression of these tumor proliferation markers differs between young and older PTC patients. Our study comprised 108 PTC patients retrospectively selected by age, i.e. those younger than 35 or older than 55 at diagnosis. Formalin-fixed, paraffin-embedded archival tissue blocks were analyzed for Bcl-2, Ki-67, and p21 protein expression by immunohistochemistry. We showed that expression of Ki-67 increases significantly with age, indicating that tumors in older patients may grow faster. This higher proliferative activity may explain the worse prognosis in these patients. Expression of p21 was higher in large tumors and in tumors extending beyond the thyroid capsule. Expression of Bcl-2 did not correlate with clinical parameters. PMID- 15756058 TI - Fluorescence cystoscopy in the management of bladder cancer: a help for the urologist! AB - As a disease characterized by a nature polymorphic and fluctuant in its evolution, superficial transitional cell carcinoma of the bladder remains a perpetual therapeutic challenge, and raises a great interest in the development of new diagnostic and surveillance techniques. This paper reviews 10 years of experience of fluorescence cystoscopy, a simple technique perfectly adapted to the current endoscopic equipment. Its principle is to enhance the visual contrast between benign and malignant cells. Three photosensitizing agents are available, two prodrugs: delta-aminolevulinic acid or hexaminolevulinate, and a natural substance: hypericin. With a detection rate of over 90% for carcinoma in situ and a real potential for detecting small tumors overlooked by standard cystoscopy, fluorescence cystoscopy may be clearly recommended in clinical practice. This technique favors a standardization of superficial bladder cancer endoscopic management and is susceptible to have a real impact on the disease recurrence and progression rate. PMID- 15756059 TI - Measurement of residual adenoma after transurethral resection of the prostate by transurethral enucleation technique. AB - OBJECTIVE: Transurethral resection of the prostate (TURP) leaves a lot of residual adenoma and has a high recurrence rate, but the residual adenoma weight has not been measured surgically. Using surgery we tried to measure the residual adenoma after TURP. MATERIAL AND METHODS: Total adenoma resection was performed via standard TURP in 64 cases with benign prostatic hyperplasia and the weight of the residual adenoma was measured by transurethral enucleation (TUE) of the prostate. RESULTS: Prostate volume averaged 37.4 ml and adenoma volume averaged 20.6 ml by TRUS. The average weight of the prostate removed by TURP was 9.8 g. After TURP residual adenoma was confirmed in all cases by TUE, and the average weight of the residual adenoma was 10.2 g. The total average prostate volume removed by TURP and TUE was 20.1 g. The average residual rate of the adenoma removed by TURP was 54.5%. CONCLUSION: TURP leaves about half of the adenoma. Therefore, TURP might be very difficult for complete resection of an adenoma. PMID- 15756060 TI - Follow-up of testicular microlithiasis for subsequent testicular cancer development. AB - INTRODUCTION: To demonstrate the relationship between testicular microlithiasis and testicular tumor development. PATIENTS AND METHODS: Between January 1996 and March 2004, bilateral testicular microlithiasis was found in 40 of the 5,263 patients who underwent scrotal ultrasonography yielding a prevalence of 0.76%. Of the 40 patients, 4 patients with concomitant testicular tumors were excluded from the study. The remaining 36 patients were enrolled into the study and followed by ultrasonography at 6-month intervals. RESULTS: Patient ages ranged between 1 and 69 years (mean 31 +/- 14 years). The median ultrasonography follow-up was 34 months (range, 1-96). Testicular tumor development was not observed in any of these 36 patients during the follow-up period. CONCLUSIONS: Extensive evaluation including computerized tomography, testicular tumor markers and testicular biopsy of patients with testicular microlithiasis is unnecessary and also increases patient anxiety. Yet annual ultrasonography and physical examination should be performed if ever until testicular microlithiasis is completely accepted as a nonpremalignant disease. PMID- 15756061 TI - Thick loop prostatectomy in the endoscopic treatment of benign prostatic hyperplasia: results of a prospective randomised study. AB - OBJECTIVE: The thick loop is a new device employed for transurethral resection of the prostate (TURP) using the standard resectoscope. The loop is broader and thicker than the standard one, resulting in better hemostasis because of its ability to cut, coagulate and vaporize tissue simultaneously. We evaluated the safety and efficacy of the thick loop device compared with the standard loop. PATIENTS AND METHODS: 103 patients with symptomatic benign prostatic hyperplasia were randomized to undergo either thick loop (51 patients) or standard loop TURP (52 patients). Patients were considered for surgery with the American Urological Association (AUA) symptom score greater than 7 and a maximum urinary flow rate <15 ml/s. Serum hemoglobin, hematocrit, electrolytes, operative time, prostate resected weight, catheterization time and complications were recorded. Twelve months later, the AUA score, maximum urinary flow rate (Q(max)) and postvoiding residual urine volume were evaluated. RESULTS: There were no significant differences between thick and standard loop TURP regarding the operative and catheterization time, prostate resection weight and postoperative levels of hemoglobin, hematocrit and electrolytes. Twelve months after TURP, the AUA score was significantly lower and Q(max) greater in the thick loop compared to the standard loop groups. CONCLUSIONS: Thick loop TURP is as safe as standard loop TURP with respect to blood loss, operative time and complications. Operating in a virtually bloodless field could allow a more radical TURP and provide an explanation for the better functional results (Q(max) and AUA score) obtained by the thick loop resection. PMID- 15756062 TI - Outpatient biofeedback relaxation of the pelvic floor in treating pediatric dysfunctional voiding: a short-course program is effective. AB - INTRODUCTION: We report the treatment results of a short course of biofeedback relaxation of the pelvic floor (BRPF) in treating children with dysfunctional voiding. MATERIALS AND METHODS: Fourteen girls and 6 boys with videourodynamically proved dysfunctional voiding were enrolled. To increase the awareness of the abnormal voiding pattern, the anatomy of the pelvis and the results of a voiding diary and videourodynamics were extensively explained to the patients/parents. Surface electromyography and uroflowmetry were used as biofeedback tools to teach the adequate relaxation of the pelvic floor during voiding. The child was asked to practice the relaxation technique at home at least twice a day. BRPF was performed once a week until the child had 2 consecutive normal bell-shaped uroflow recordings. Patients were reevaluated at 4, 8, 12 and 24 weeks after BRPF training. RESULTS: The mean patient age was 8.3 +/- 3.8 years. The mean number of BRPF training sessions was 2.2 +/- 0.9. The mean follow-up period was 18.9 months. Normalization of abnormal uroflowmetry was achieved in 18 patients (90%). The mean maximal uroflow rate and voided volume increased from 13.3 +/- 4.3 to 18.0 +/- 3.4 ml/s (p < 0.01) and from 138 +/- 56 to 193 +/- 65 ml (p < 0.01), respectively. The postvoiding residual urine decreased from 54.5 +/- 47.6 to 21.3 +/- 10.6 ml (p < 0.01). Complete and partial resolution of voiding symptoms was achieved in 14 (70%) and 6 patients (30%), respectively. Recurrence was noted in 2 of the 10 patients who had complete resolution of symptoms and who had been followed up for more than 6 months. CONCLUSIONS: Short-course outpatient BRPF is an effective treatment of pediatric dysfunctional voiding. PMID- 15756063 TI - Lower urinary tract reconstruction in ectopic ureteroceles. AB - OBJECTIVES: The management of ectopic ureteroceles is a challenging entity in pediatric urology. In our study, we aimed to determine the outcome after lower urinary tract reconstruction in ectopic ureteroceles. MATERIALS AND METHODS: A total of 18 (12 girls, 6 boys) children with ectopic ureteroceles, treated between 1993 and 2003 by complete reconstruction, were enrolled in the study and their records were retrospectively reviewed. RESULTS: Ureterocele was found to be unilateral in 16 and bilateral in 2 children. Four patients had been previously managed by endoscopic interventions and 1 patient underwent partial nephrectomy. Ureterocelectomy was performed on 20 renal units, and of these 20 renal units, 9 underwent heminephroureterectomy, 1 ureterectomy, and 3 (with single system ureteroceles) nephroureterectomy. Seven (35%) renal units with adequate function underwent ureteral reimplantations. After a mean follow-up of 5.4 years (range 4 months to 9.5 years), no patient required a second procedure. Contralateral reflux that developed in 2 children after surgical treatment resolved spontaneously after 1 year of follow-up. CONCLUSION: In the management of ectopic ureterocele, lower urinary tract reconstruction is an effective treatment alternative. According to the functional status of the renal parenchyma involved, the surgical procedure was limited to the lower urinary tract in 35% of the cases. PMID- 15756064 TI - Advantages of video-assisted approach in detecting epididymal anomalies and treatment of nonpalpable testis. AB - In children with a nonpalpable abdominal testis, preoperative localization is very helpful prior to surgical investigation both to reduce the time required and to plan the correct surgical procedure. It is no doubt that laparoscopy is useful in both diagnosis and management of nonpalpable testis. In this study, 15 patients with 17 nonpalpable testes admitted to our clinic in 2 years period were evaluated retrospectively. Ultrasound study was performed in all patients after detailed physical examination. Testicular location, size, and configuration of epididymis were determined by laparoscopy and compared with ultrasound findings. Laparoscopy, in conjunction with ultrasound, permits not only a better localization and anatomy of the testis but is also superior in planning the best choice of management. We believe that the video-assisted inguinal approach is a feasible method in the treatment of nonpalpable testis when compared to pure open surgery. PMID- 15756065 TI - Trigonoplasty to treat secondary vesicoureteral reflux in neurogenic bladders. AB - OBJECTIVES: The effectiveness of Gil-Vernet's trigonoplasty, with technical modifications, was studied in treating vesicoureteral reflux (VUR) in neurogenic bladders. METHODS: Modified Gil-Vernet's trigonoplasty, making the transmural ureters advance over the midline and cross each other in the trigone, was applied in patients with neurogenic bladder dysfunction and VUR, diagnosed by voiding cystourethrogram. Most of the patients were sufficiently followed up by cystourethrogram. RESULTS: 26 patients with neurogenic bladder dysfunction were diagnosed as having 43 refluxing units and underwent modified Gil-Vernet's trigonoplasty. Reflux was grade I in 5 units, grade II in 7, grade III in 5, grade IV in 18, and grade V in 8, with unilateral reflux in 9 patients and bilateral reflux in 17. Surgery was successful in 95.3% of 43 refluxing units. 22 patients were followed 3-6 months after the operation, and the cure rate was 90.9% (no reflux under voiding cystourethrogram). 18 patients were followed up for more than 2 years without recurrence of VUR. CONCLUSIONS: Modified Gil Vernet's trigonoplasty might be a useful technique in the management of patients with VUR secondary to neurogenic bladder dysfunction. PMID- 15756066 TI - Endoscopic treatment of ureteric strictures: acucise, cold-knife endoureterotomy and wall stents as a salvage approach. AB - OBJECTIVE: To evaluate the factors influencing the results of endoureterotomy using cold-knife and cutting balloon dilatation, and permanent ureteral wall stents in patients with benign ureteral strictures after different operations affecting the ureter. MATERIALS AND METHODS: Over a 4-year period, in 18 patients, endoscopic cold-knife and Acucise endoureterotomies were performed in 13 and 7 renal units, respectively. Eight Memoterm permanent ureteral wall stents were inserted into 7 patients when endoureterotomy failed. Successful outcome was defined by the absence of re-stricture assessed both clinically and radiologically. RESULTS: The strictures were secondary to ureterolithotomy in 6, ureteroscopy in 3, gynecological procedures in 4, abdominal surgeries in 2, transplantation in 2 and continent urinary diversion in 1. The right and left ureters were unilaterally affected in 5 and 11 patients, respectively (5 of them had a solitary kidney), while the remaining 2 patients had bilateral ureteral strictures. We achieved total ureteral patency of 3 (43%) and 7 (54%) renal units with Acucise and cold-knife incision, respectively. Obstructive uropathy was resolved in 6 renal units (75%) of 8 using ureteral wall stents. CONCLUSION: Endoureterotomy with cold-knife or Acucise cutting balloon dilatation is effective in the treatment of iatrogenic ureteral strictures, but only in a selected group. Based on our results, the favorable prognostic criteria for endoureterotomy are the length (< or =1.5 cm), the nonischemic nature of the stricture and adequate renal function. As a salvage approach, permanent self expanding ureteral wall stents with a 75% success rate may provide a satisfactory outcome for decompression of an obstructed system. PMID- 15756067 TI - Long-term surgical results of pubovaginal sling procedure using polypropylene mesh in the treatment of stress urinary incontinence. AB - INTRODUCTION: Surgery for stress urinary incontinence (SUI) in women might cure incontinence symptom but develop new problems. This study assessed the long-term outcome of pubovaginal sling procedure using polypropylene mesh in patients with SUI. MATERIALS AND METHODS: A total of 108 patients with various types of SUI received pubovaginal sling procedure using polypropylene mesh. A surgical technique that did not create bladder outlet obstruction after operation was used in all the patients. Video urodynamic study was performed at baseline, 2 weeks and 3 months after operation. The long-term surgical results and patients' satisfaction were assessed. RESULTS: The age of patients ranged from 33 to 94 years (mean 62.6 +/- 12.0 years). The follow-up period ranged from 24 to 72 months (median 46 months). Among the 108 patients, a successful result was noted in 92 (85.2%), including a continence rate of 65.7% and mild SUI in 19.4% of patients. Treatment failure was noted in 16 patients (14.8%), including 13 (12%) with urge incontinence and 3 (2.8%) with severe SUI that was cured by a second sling. There were 104 patients (96.3%) who could void volitionally with little residual urine, whereas 4 (3.7%) needed transvaginal urethrolysis. Urodynamic study revealed no significant changes in the mean values of parameters at 3 months after operation. However, 22 (20%) patients had an increase in voiding pressure by 50% at 3 months postoperatively. The reported satisfactory rate was 89.8%. CONCLUSIONS: Pubovaginal sling procedure using polypropylene mesh had a high success rate in all types of SUI. The continence rate was suboptimal, indicating that the absence of bladder outlet obstruction might result in inadequate urethral compression after pubovaginal sling procedure. PMID- 15756068 TI - Bladder carcinoma during pregnancy. AB - INTRODUCTION: We report 3 cases of bladder cancer during pregnancy and give a review of the literature in an attempt to evaluate tumor at presentation, characteristics, maternal and fetal outcome. MATERIALS AND METHODS: The case history of 3 pregnant women treated for bladder cancer in 2001 together with the results of a MEDLINE search from 1966 to 2003. RESULTS: Out of 27 cases of nonbilharzial bladder carcinoma, 74% presented with transitional cell carcinoma. Five patients had muscle-invasive tumors. Major symptom was hematuria in 81%, which was initially mistaken as vaginal bleeding in 22%. Only half of the tumors were identified by ultrasonography. Although superficial bladder carcinoma was transurethrally resected alone, outcome and prognosis are good. But the prognosis of locally advanced bladder carcinoma is poor. None of the fetuses delivered before 30 weeks of gestation survived. Two of the 5 patients died from the disease and follow-up is only short in the rest. CONCLUSION: Any doubtful genital bleeding during pregnancy without definite proof of vaginal/cervical origin should be investigated by both ultrasonography of the upper urinary tract and urethrocystoscopy. Superficial bladder tumors can be most effectively treated by transurethral resection followed by cystoscopy, whereas the prognosis of muscle invasive bladder carcinoma is poor and demands more radical treatment, depending on the stage of pregnancy. PMID- 15756069 TI - Rectal distention inhibits the spinal micturition reflex via glycinergic or GABAergic mechanisms in rats with spinal cord injury. AB - BACKGROUND: We examined the influence of rectal distention on the spinal micturition reflex and the mechanism underlying its inhibition of bladder contraction. METHODS: Fourteen conscious female Sprague-Dawley rats were used in this study after transection of the lower thoracic cord. Isovolumetric cystometry was performed before and after distention of the rectum by inflation of a rectal balloon, followed by intrathecal injection of strychnine (a selective glycine receptor antagonist) or bicuculline (a GABA(A) receptor antagonist) into the lumbosacral cord. RESULTS: Rectal distention (1.0-3.0 cm(3)) prolonged the interval, decreased the amplitude, and shortened the duration of bladder contraction, and eventually almost abolished bladder activity. After intrathecal injection of strychnine (0.001-10 microg) or bicuculline (0.001-1 microg) in animals with inhibition of bladder activity by rectal distention, the interval, amplitude, and duration of bladder contraction returned to baseline. CONCLUSION: These results suggest that there is an inhibitory rectovesical reflex in the lumbosacral cord of rats with spinal cord injury, which modulates the spinal micturition reflex via glycinergic or GABAergic mechanisms. PMID- 15756070 TI - Effects of early phase of preconditioning on rat testicular ischemia. AB - INTRODUCTION: Brief episodes of ischemia followed by periods of reperfusion generate a powerful protective mechanism in cell, tissue or organ, which increase the resistance to further ischemic damage. This is known as ischemic preconditioning, and has not been investigated in testis. The present experiments were undertaken to determine whether early phase of ischemic preconditioning is evident in rat testis. MATERIALS AND METHODS: Surgery was conducted under thiopental (60 mg/kg, intraperitoneal) anesthesia in male Wistar rats. Surgical procedures were performed through a midline incision. Group 1 was designed as a sham group. In group 2, which served as the ischemia group, the animals were subjected to unilateral testicular torsion by rotating the left testis 720 degrees in a clockwise direction. Then, this testis was maintained in the torsion position by fixing with a silk suture to the scrotal wall for 90 min. In groups 3 and 4, 5 or 10 min ischemia followed by 10 min reperfusion was introduced, respectively, to induce single cycle ischemic preconditioning. In group 5, which served as the multiple cycle preconditioning group, 3 cycles of 10 min ischemia and 10 min reperfusion were applied prior to 90 min ischemia. Both ipsilateral and contralateral testes were removed from the rats at the end of the experimental periods, and tissue malondialdehyde (MDA), nitric oxide (NO) levels, xanthine oxidase (XO), myeloperoxidase (MPO) and superoxide dismutase (SOD) activities were measured. Both testes were also evaluated histologically, assessing interstitial edema, congestion, hemorrhages, rupture of tubules and Leydig cell proliferation. RESULTS: 90 min ischemia produced a marked increase in MDA level in left testis. However, all ischemic preconditioning protocols used in this study did not show any significant modification in MDA, NO levels or XO, MPO and SOD activities. Histological grading scale was also similar in ischemia and preconditioning groups. CONCLUSION: These results suggest that there are no protective effects with ischemic preconditioning in rat testis as showed by biochemical analysis and histological examinations. PMID- 15756071 TI - Effects of micronised purified flavonoid fraction on pain, spermiogram and scrotal color Doppler parameters in patients with painful varicocele. AB - PURPOSE: Micronised purified flavonoid fraction (MPFF) is a phlebotropic drug improving venous tone, elasticity and lymphatic drainage, decreasing venous distensibility, venous emptying times, reflux time of venous blood and capillary hyperpermeability, and having antioxidant activities. Since varicocele is a venous pathology, we evaluated the effects of MPFF on pain, spermiogram and color Doppler parameters in patients with painful varicocele. MATERIAL AND METHODS: Semen analyses and Doppler sonography were performed in 16 patients (aged 20-62 years, mean 31.2 +/- 10.9) before and after 1,000 mg/day MPFF treatment. Pain change was assessed with visual analogue scale scored between 0 and 10. Patient satisfaction with outcome and side effects were recorded. RESULTS: Mean pain scores at 1, 3, and 6 months were 1.25 +/- 1.34, 0.25 +/- 0.68, and 0.12 +/- 0.34, respectively, all being significantly lower (p < 0.001 for each) than baseline (4.93 +/- 1.77). Mean score at 12 months (0.93 +/- 1.06) was still lower than baseline (p < 0.001) but higher than at 6 months (p = 0.019). Respectively, 14 (87.5%) and 2 (12.5%) patients reported complete and very significant resolution of pain at the 6th month. All patients were very satisfied with the outcome. While semen volume, total sperm count, sperm concentration and morphology did not change significantly, motility increased significantly (p = 0.009) due to decrease of grade 0 sperms (p = 0.006) and increase of grade 3 + 4 sperms (p < 0.001). Reflux time of left spermatic vein during the Valsalva maneuver decreased significantly (p = 0.003). No significant change occurred in reflux time of the right spermatic vein. No side effect was recorded. CONCLUSIONS: Results of this preliminary study suggest the safety and efficacy of MPFF in the treatment of varicocele-associated pain. However, no recommendation for the use of MPFF in the treatment of pain associated with varicocele can be made before these preliminary results are confirmed by a randomized placebo controlled trial. PMID- 15756072 TI - Prostate fossa packing: a simple, quick and effective method of achieving hemostasis in suprapubic prostatectomy. AB - INTRODUCTION: Suprapubic prostatectomy remains a very common surgical treatment for bladder outlet obstruction from an enlarged prostate in the developing world. It is a technically simple procedure which can be learned by nonurologists who perform the bulk of these procedures in the developing world. Intraoperative and postoperative bleeding, however, can be a problem. We describe our experience with prostatic fossa packing at the time of the procedure, as a very simple, quick and effective technique for maximal hemostasis. MATERIAL AND METHODS: We have used this technique for several years at our institution and the results have been excellent. However, the major advantages of this technique became highlighted when the primary author visited Eritrea on a medical mission trip and used it in 42 consecutive cases. We looked at the hemorrhagic complications, both intraoperatively and postoperatively, infections and other complications related to the technique. RESULTS: The 42 patients were typical of the patients that are seen in the developing world for suprapubic prostatectomy. The age range was 49 102 years. 75% had indwelling urethral catheters secondary to urinary retention. The duration of the catheters was from 2 weeks to 10 months. All the patients who had the indwelling catheters had bacteriuria. 56% had associated bladder calculi and 25% had associated inguinal hernias. The average surgical time was 45 min. The average intraoperative blood loss was estimated at 160 ml. There were no blood transfusions. There were no infections and there were no complications related to the packing. CONCLUSION: Packing of the prostatic fossa at the time of enucleation of the adenoma is a very simple, quick and effective way to achieve hemostasis in suprapubic prostatectomy. It is also very easy to learn. With this technique we have achieved zero rate of transfusion and no infections. The surgical time is also reduced. We have had no morbidities associated with this technique and highly recommend it. PMID- 15756073 TI - Urethral advancement procedure for reconstruction after excision of male parameatal melanoma in situ. AB - We successfully reconstructed the glans penis and external meatus by means of the urethral mobilization and advancement technique for an adult patient with a parameatal lesion of melanoma in situ. PMID- 15756074 TI - Fulminant hepatic failure secondary to metastatic prostate cancer. AB - The most common sites of metastatic disease from prostate cancer include the bones, lymph nodes and less commonly the lungs, adrenal glands, brain and kidneys. Acute fulminant hepatic failure secondary to cancer metastasis is rare and has unique clinical presentation. We describe a case of fatal liver failure in a 71-year-old male due to metastases to the liver from a prostatic adenocarcinoma. Thus in patients with metastatic cancer and elevated liver function tests, the possibility of hepatic involvement by the cancer should be considered as a possible cause of hepatic failure. PMID- 15756075 TI - Bilateral perirenal lymphangioma: a conservative approach. AB - The lymphangioma is a well-recognized benign tumor most frequently encountered in infancy and childhood. The kidneys are rarely involved. Despite its benign nature, most of the patients were treated by partial or radical nephrectomy. We describe a case of bilateral perirenal lymphangioma in a 33-year-old man for whom we chose a conservative approach. PMID- 15756076 TI - Transverse testicular ectopia associated with persistent Mullerian duct syndrome. A case report. AB - Transverse testicular ectopia (TTE) associated with persistent Mullerian duct syndrome (PMDS) is a rare genitourinary anomaly. The clinical and operative findings and treatment are discussed. It is very important to perform a careful exploration in TTE when the testes are undescended, in order to exclude the presence of PMDS. Transseptal orchidopexy is the surgical treatment of choice. PMID- 15756077 TI - Emergency medicine research. PMID- 15756078 TI - Management of clinical fractures of the scaphoid: results of an audit and literature review. AB - OBJECTIVES: Two to 5% of scaphoid fractures are missed on initial presentation. The failure of early recognition and treatment are considered to contribute to delayed union and non-union. Despite advances in diagnostic imaging, a dogmatic approach has persisted in the management of patients with clinical suspicion but no radiographic evidence of scaphoid fracture. A critical analysis of the current treatment protocol of indiscriminate cast immobilization and serial clinical and radiographic follow-up is presented. METHODS: A prospective study involving 90 patients with clinical signs suggestive of scaphoid injury, followed up until a definite boney injury was demonstrated or the patient was discharged. A review of the literature was conducted to question the need for immobilization in these patients and the potential use of other forms of diagnostic imaging in screening for occult scaphoid fractures. RESULTS: The incidence of true fractures of the scaphoid was 6.66% (5/75). Ten patients (13.33%) had other injuries around the wrist unrelated to the scaphoid. Eighty per cent of the patients had no definite boney injury and were needlessly immobilized, and followed up. A total of 128 scaphoid casts, 135 sets of scaphoid X-rays, 135 clinic appointments and a cumulative 148 weeks of cast immobilization involved patients with normal wrists. CONCLUSION: The incidence of radiologically inapparent fractures of the scaphoid is low. The use of a tender anatomical snuff box as the only clinical sign in the diagnosis of scaphoid injury is unsatisfactory. Other injuries around the wrist must be carefully excluded. There is insufficient evidence to support immobilizing all patients with clinical scaphoid fractures. For suspected fractures with no radiological evidence, symptomatic treatment is probably sufficient. Most occult fractures are visible at 2 weeks. Both magnetic resonance imaging and bone scintigraphy are accurate and cost effective and should be performed earlier rather than later. PMID- 15756079 TI - Management of new-onset atrial fibrillation in the emergency department: is there any predictive factor for early successful cardioversion? AB - OBJECTIVES: The aim of this retrospective study was to assess the initial management of atrial fibrillation (AF) in the emergency department (ED) and to identify predictive factors of early conversion to sinus rhythm, which could justify a short stay in the ED observation unit (EDOU) instead of longer hospitalization. PATIENTS AND METHODS: All patients with acute AF, either of new onset or recurrent, admitted to our hospital during a 12-month period were included in the study. Hospital records were reviewed retrospectively. The success of conversion to sinus rhythm was recorded in association with a series of clinical and laboratory factors. RESULTS: Sixty-seven patients (39 men and 28 women), with a mean age of 63.6+/-12.2 years, were studied. The most frequent presenting symptom was palpitations (n=40, 59.7%). In forty-two patients (62.7%) the duration of symptoms was less than 48 h. Digoxine was the anti-arrhythmic agent most frequently administered (n=26, 38.8%), followed by amiodarone (n=17, 25.4%). Fifty patients (73.1%) converted to sinus rhythm and for 45 of them conversion took place during their stay in the ED or in the EDOU. Factors associated with early conversion to sinus rhythm were aged younger than 65 years (P=0.021) and symptom duration of less than 48 h (P=0.001). On the other hand, the presence of signs of heart failure was significantly associated with unsuccessful early cardioversion (P=0.001). CONCLUSIONS: The majority of patients admitted with AF of acute onset had early conversion to sinus rhythm. AF in young patients, with a duration of symptoms of less than 48 h and without signs of heart failure can be managed in the EDOU, thus avoiding a longer hospitalization. PMID- 15756080 TI - Improving the pattern of work towards emergency medicine at the Emergency Department in Rambam Medical Centre. AB - BACKGROUND: Rambam Medical Centre is a 950-bed referral hospital, containing 34 beds in the Emergency Department (ED). The arrangement of the ED was based on two separate units: trauma and medical. Patient evaluation lasted many hours, with an abuse of consultants, laboratory and imaging services. OBJECTIVE: To describe the improvement of patient management in our ED by changing working patterns according to emergency medicine guidelines. METHODS: The trauma and medical units were combined into a one-unit ED, managed by one director and two specialists in emergency medicine. The goal of the ED was defined as the treatment and triage of patients towards discharge or admission in the minimal time needed. New protocols for clinical evaluation, blood and imaging tests, consultation facilities and pain management were implemented according to emergency medicine practice. Once a decision for admission is made, a computerized program assigns the patient to the appropriate ward. RESULTS: There was a dramatic reduction (44%) in the time needed for patient evaluation. The number of patients waiting more than 4 h and 8 h for admission was reduced significantly (11 versus 38% and 1 versus 24%, respectively). The total number of blood tests was reduced by 45%, and the number of blood and urine cultures by 81 and 87%, respectively. Two years after the change, the ED won first place in patient satisfaction screening. CONCLUSIONS: By changing work methods according to emergency medicine guidelines, and by using protocols written for emergency medicine, a significant improvement in our duties as an ED was achieved. The backing of the hospital management helped to implement the changes, which led to the functional improvement. PMID- 15756081 TI - Adverse events and death in stroke patients admitted to the emergency department of a tertiary university hospital. AB - OBJECTIVES: To identify the occurrence of adverse events in stroke patients presenting to the emergency department of a tertiary university facility, and to disclose the categories of adverse events associated with death. METHODS: This matched case-control study enrolled 468 patients admitted with stroke to the emergency department from March 1996 to September 1999. The cases comprised 234 consecutive deaths and the controls 234 discharged patients, matched for primary diagnosis and admission period. Adverse events, detected by chart review, were classified according to the degree of severity, immediate causes, and professional category. The association with death was analysed by conditional logistic regression. RESULTS: Adverse events totaled 1218 and occurred in 295 patients: 932 events (76.5%) in 170 cases and 286 (23.5%) in 125 controls. Major adverse events equaled 54.1% of all events (659 episodes): 538 events in 143 cases and 121 in 65 controls. Diagnostic or therapeutic procedures and nursing activities accounted for 55.2% of events. Nursing (38.4%) and medical (31%) adverse events represented the most common related professional categories. A significant association with death was found for major adverse events, medical adverse events, and nosocomial infections, with adjusted odds ratio estimates of 3.74 [95% confidence interval (CI) 1.64-8.54], 3.71 (95% CI 1.61-8.53), and 3.22 (95% CI 1.21-8.59), respectively. CONCLUSION: Adverse events, mostly severe, predominated among deceased patients, resulting mainly from diagnostic or therapeutic procedures and nursing activities. In spite of limitations concerning the observational retrospective nature of this study, we found that severe adverse events, medical adverse events, and nosocomial infections were significantly associated with death in stroke patients. PMID- 15756082 TI - An analysis of spoken language expression during simulated emergency call triage. AB - OBJECTIVES: Volunteer citizens were recruited to perform simulated emergency calls, and the expressions and content of these telephone calls were analysed to examine risk factors associated with the success or failure of communication. SUBJECTS AND METHODS: Six physicians played the role of patients who had various symptoms, such as cerebral stroke and ischaemic heart disease. Eighty-four volunteer citizens made simulated emergency calls. Physicians at a simulated call centre communicated with each caller regarding the patient's body position, respiratory condition, and cardiovascular status. Details of the telephone communications were analysed to determine if communication was successful. RESULTS: Telephone communications that resulted in the correct understanding of a simulated patient's condition were as follows: 60.2% of sessions (32/50) on whether or not a patient was breathing; 47.8% of sessions (22/46) on whether or not a patient had a pulse (carotid or radial artery); and 86.2% of sessions (56/65) on patient body position. How a simulated dispatcher verbally expressed questions was the most influential factor in the success of communication regarding respiratory condition and body position. Avoiding vague language, giving specific instructions for checking a patient, and finally reminding the caller to perform the explained procedures led to a high rate of successful communications. Various spoken expressions by simulated dispatchers in confirming patient pulse did not have any impact on the success or failure of communications. CONCLUSION: In developing a 'protocol for emergency call triage' to achieve a high rate of successful emergency communications, an analysis of expressions using simulated patients is useful. PMID- 15756083 TI - Pyridoxine in clinical toxicology: a review. AB - Pyridoxine (vitamin B6) is a co-factor in many enzymatic pathways involved in amino acid metabolism: the main biologically active form is pyridoxal 5 phosphate. Pyridoxine has been used as an antidote in acute intoxications, including isoniazid overdose, Gyromitra mushroom or false morrel (monomethylhydrazine) poisoning and hydrazine exposure. It is also recommended as a co-factor to improve the conversion of glyoxylic acid into glycine in ethylene glycol poisoning. Other indications are recommended by some sources (for example crimidine poisoning, zipeprol and theophylline-induced seizures, adjunct to d penicillamine chelation), without significant supporting data. The value of pyridoxine or its congener metadoxine as an agent for hastening ethanol metabolism or improving vigilance in acute alcohol intoxication is controversial. This paper reviews the various indications of pyridoxine in clinical toxicology and the supporting literature. The potential adverse effects of excessive pyridoxine dosage will also be summarized. PMID- 15756084 TI - Delayed pericardial tamponade after penetrating chest trauma. AB - Delayed pericardial tamponade (PT) after penetrating heart trauma is now a rare condition as a result of advances in medical and surgical management. We report the case of a 32-year-old man with delayed PT after a stab wound from a knife. The initial evaluation was consistent with a traumatic apical myocardial infarction. After an uneventful initial course, the patient developed acute PT, which required emergency surgery. A thrombus was discovered over a laceration in the mid-segment of the left anterior descending artery and a simple suture was performed. PMID- 15756086 TI - A case of catamenial pneumothorax caused by thoracic endometriosis. PMID- 15756085 TI - Delayed perforation of right ventricle with cardiac tamponade: a complication of pacemaker implantation. AB - We report a case of delayed cardiac tamponade that resulted from pacemaker implantation in the emergency room of a large urban hospital. A 19-year-old male patient with a pacemaker implanted 10 days earlier suffered from delayed perforation of the right ventricle with cardiac tamponade. A review of the literature revealed that cardiac tamponade is a rare complication of pacemaker implantation. Pacemakers should only be implanted by physicians with relevant experience, and emergency room physicians should be aware of the possibility of the associated complications and be prepared to treat them. PMID- 15756087 TI - Thrombolytic therapy during prolonged in-hospital cardiac arrest. AB - We report a case of cardiac arrest secondary to acute myocardial infarction that was successfully resolved after thrombolytic therapy. This case suggests that thrombolysis can be used safely also during prolonged cardiopulmonary resuscitation for in-hospital cardiac arrest. PMID- 15756088 TI - Clinical diagnosis of fractures in a paediatric population. AB - Diagnosing fractures in the paediatric population is a problematical process for which there are currently no accepted clinical criteria. We studied the physical signs sought by accident and emergency staff in 126 children with suspected fractures. We found a significant correlation between 'point tenderness' and fracture, as demonstrated by plain radiograph. 'Swelling' and 'redness' approached significance. These results correlate well with previous work in this area, but further research using a larger sample is required as confirmation. PMID- 15756089 TI - Organophosphate poisoning: the lesser-known face of a toxidrome. PMID- 15756090 TI - When is a herb a drug? AB - With the growing popularity and use of herbal remedies in the industrial countries there has been an increasing recognition and reporting of adverse reactions. Drug development represents discovery and characterization of naturally occurring plant metabolites or their synthetic analogues. Therefore, a herbal remedy should be considered a drug for all clinical purposes. The liver, being central to the metabolism of virtually all xenobiotics, is inevitably exposed to reactive metabolites and liver injury is a potential complication of nearly every medication, including herbal remedies. Recognition of herb induced hepatotoxicity depends on clinical awareness of the problem, a suspicion that a herbal preparation is involved, and detailed enquiry regarding the use of all supplements and remedies. Herbal medicines should be regulated more systematically and monitoring for adverse reaction should improve. PMID- 15756091 TI - Crohn's disease patient characteristics in a tertiary referral center: comparison with patients from a population-based cohort. AB - OBJECTIVES: Data on the clinical presentation, effects of therapy and prognosis of patients with Crohn's disease are often based on patients from specialized referral centers. We assessed the extent of the selection based on the clinical and demographic characteristics. METHODS: All patients with Crohn's disease presenting to the University Hospital of Regensburg (Medical Department) were analyzed retrospectively with respect to demographic and disease specific characteristics. Only patients diagnosed <2 years before presentation were included in the main analysis. The original data from a population-based, prospectively assembled incidence cohort were available for comparison (EC-IBD, northern centers only, n=475). Age at diagnosis, disease location and behavior were categorized according to the Vienna classification. Differences were examined using chi-square tests. MAIN RESULTS: At the referral center, 394 patients were treated within a 5-year period. Of these, 116 patients fulfilled the inclusion criteria for the comparative analysis. Sixteen percent of the referral patients were diagnosed at age 40 or older, as compared with 32% in the population-based group (P<0.004). The distribution of disease location, sex, smoking behavior and positive family history was similar in both groups. Among the referral patients, more had fistulas (39% versus 20%, P<0.001). Also, more patients were receiving steroids (49% versus 27%) or other immunosuppressive therapy (12% versus 4%). The selection effects increase with duration of disease. CONCLUSIONS: Patients with late onset of disease, inflammatory only disease behavior and no need for immunosuppression are under-represented at a tertiary referral center. PMID- 15756092 TI - Complications of portal hypertension in adults: a French consensus. AB - Portal hypertension is responsible for different severe complications such as variceal bleeding, ascites and pulmonary disorders. Different pharmacological and endoscopic treatments have been proposed but some recommendations may differ. The conclusions of a French consensus conference on the prevention and treatments of the main complications of portal hypertension in adults are reported. PMID- 15756093 TI - Impact of irritable bowel syndrome on patients' lives: development and psychometric documentation of a disease-specific measure for use in clinical trials. AB - OBJECTIVE: To develop a disease-specific questionnaire to capture the impact of irritable bowel syndrome (IBS) and its treatment on patients' lives, the Irritable Bowel Syndrome Impact Scale (IBS-IS). PATIENTS AND METHODS: One hundred and fifty-five IBS patients participated (126 (81%) female; age (mean+/-SD) 45.5+/-12.4 years). We developed the initial 39 items from the literature, available IBS-specific instruments and input from physicians, nurses and patients. We deleted IBS-IS items with a high ceiling effect, items that measured a different construct and items showing a high correlation (r>0.90) with another item and with Rasch analysis, leaving 26 items. We then applied exploratory factor analysis to examine domain groupings. Subjects completed the IBS-IS instrument, the Gastrointestinal Symptom Rating Scale for IBS (GSRS-IBS), Short Form-36 (SF-36), Visceral Sensitivity Index (VSI), and Hospital Anxiety and Depression (HAD) scale. Internal consistency, construct validity and discriminate validity were assessed. RESULTS: The 26 items represented five domains: fatigue, impact on daily activities, sleep disturbance, emotional distress and eating habits. The internal consistency reliability for the domains was 0.87 to 0.96. Most associations between similar constructs in the IBS-IS, GSRS-IBS, SF-36, VSI, and HAD were >0.40. Each IBS-IS domain score decreased with increasing IBS symptom severity (P<0.05), and the patients scored >5 score units lower than a US general population scored on all eight SF-36 dimensions. CONCLUSION: The IBS-IS is a short, user-friendly instrument with excellent psychometric properties that has potential usefulness for clinical trials. PMID- 15756094 TI - Tegaserod is safe, well tolerated and effective in the treatment of patients with non-diarrhoea irritable bowel syndrome. AB - OBJECTIVE: To evaluate the safety/tolerability and efficacy of tegaserod, a 5-HT4 receptor partial agonist, in the treatment of patients with non-diarrhoea irritable bowel syndrome (non-D-IBS) in Switzerland. METHODS: This was an 8-week, open-label, prospective, multicentre study. Patients (> or =18 years old) met the Rome II diagnostic criteria for IBS, excluding those with diarrhoea for > or =14 days in the previous 3 months. Details of IBS symptoms experienced in the preceding week were recorded at visit 1 (day 1). Eligible patients received 6 mg tegaserod twice daily for 8 weeks. Adverse events (AEs) and serious AEs were recorded, along with detailed assessment of diarrhoeal episodes. Efficacy assessments included the overall number and percentage of responders after 8 weeks' treatment. RESULTS: A total of 850 patients (72% women; mean age, 51.4 years) were enrolled, and 843 received at least one dose of tegaserod. AEs were reported in 38% of patients, of which 13% were drug-related. Diarrhoea occurred early during treatment (13% in the first week, 7% thereafter), was mild to moderate in severity, was transient and was resolved with continued treatment. In total, 208 patients left the study early, primarily due to AEs. Diarrhoea accounted for 68 of these discontinuations. Nine serious AEs were reported but these were not related to tegaserod treatment. Sixty-six percent of patients responded to tegaserod on the Subject's Global Assessment of relief after 8 weeks. Benefits were also seen across individual IBS symptoms. CONCLUSION: Tegaserod (6 mg twice daily) appears to be safe, well-tolerated and effective in the treatment of non-D-IBS over 8 weeks. PMID- 15756095 TI - Localization of gastrinomas by selective intra-arterial calcium injection in patients on proton pump inhibitor or H2 receptor antagonist therapy. AB - Preoperative localization is important for the successful treatment of gastrinomas. The aim of this study was to investigate whether selective intra arterial calcium injection and hepatic venous sampling was able to localize gastrinomas in four patients who remained on proton pump inhibitor or H2 antagonist therapy. Calcium gluconate was injected directly into the arteries supplying the pancreas and liver after standard selective angiography. Gastrin levels were then measured in samples taken from the right hepatic vein. Calcium gluconate produced a diagnostic rise (at least 2-fold) in serum gastrin and unequivocally localized the tumour to a specific vascular territory in each case. One patient did not undergo surgery. In the remaining three patients, surgery confirmed the position and histology of the tumour. This study shows that in four patients with a confirmed gastrinoma we were able to localize the gastrinoma by selective intra-arterial calcium injection and hepatic venous sampling, whilst the patients remained on proton pump inhibitor or H2 antagonist therapy, thereby reducing their risk of gastrointestinal perforation. PMID- 15756096 TI - A solitary tuberculous ulcer in the oesophagus. AB - A 48-year-old woman born in Pakistan was evaluated for dysphagia. Endoscopy showed a solitary ulcerative oesophageal lesion. Cultures were positive for mycobacterium tuberculosis. Additional imaging showed no other manifestations of tuberculosis. Oesophageal tuberculosis is a rare entity, especially as a primary manifestation defined as involvement of the oesophagus without signs of disseminated disease. Therefore, this case was classified as primary oesophageal tuberculosis. PMID- 15756097 TI - Idiopathic gastric acid hypersecretion presenting as a diarrheal disorder and mimicking both Zollinger-Ellison syndrome and Crohn's disease. AB - Many gastric acid hypersecretory states (basal acid output of greater than 15.0 mEq/h) exist for which the etiology is known, such as Zollinger-Ellison syndrome, systemic mastocytosis, antral exclusion, antral predominant Helicobacter pylori gastritis (antral G cell hyperplasia), chronic gastric outlet obstruction, short gut syndrome and basophilic leukemias. However, many hypersecretory patients have no identified etiology for their acid hypersecretion and are designated as idiopathic gastric acid hypersecretors with a basal acid output of greater than 10 mEq/h and a normal serum gastrin level. Because of the gastric acid hypersecretion these patients also commonly have an increased frequency of stools. Idiopathic gastric acid hypersecretion represents a known cause of gastric acid hypersecretion that is far more common than Zollinger-Ellison syndrome and it has a markedly different treatment regimen and natural history. We report a case of a patient with idiopathic gastric acid hypersecretion previously misdiagnosed as having Crohn's disease because of a presenting complaint of diarrhea and mimicking Zollinger-Ellison syndrome because her fasting serum gastrin level was elevated when incorrectly measured in the presence of antisecretory treatment. PMID- 15756098 TI - Herbal hepatotoxicity: acute hepatitis caused by a Noni preparation (Morinda citrifolia). AB - A 45-year-old patient was sent to our department because of highly elevated transaminases and elevated lactate dehydrogenase. His medical history was unremarkable and he took no medication on regular basis. Physical examination did not detect any abnormalities. There was no evidence for viral hepatitis, Epstein Barr virus or cytomegalovirus, autoimmune hepatitis, Budd-Chiari syndrome, haemochromatosis or Wilson's disease. During the interview he admitted that for 'prophylactic reasons' he had been drinking the juice of Noni (Morinda citrifolia), a Polynesian herbal remedy made from a tropical fruit, during the preceding 3 weeks. This gave rise to the suspicion of herbal toxicity, which was confirmed by a liver biopsy. After ceasing the ingestion of Noni, transaminase levels normalized quickly and were within normal ranges 1 month after the first presentation. To our knowledge, this is the first report of hepatotoxicity caused by this herbal remedy, which has been highly praised in the tabloid press. PMID- 15756099 TI - Cerebellar degeneration and hearing loss in a patient with idiopathic myenteric ganglionitis. AB - A 35-year-old male with an 11-year history of intestinal pseudo-obstruction associated with an idiopathic inflammatory insult of the myenteric plexus and the presence of circulating anti-Hu antibodies developed a neurological syndrome characterized by bilateral hearing loss, deteriorating balance, an unsteady gait and difficulty in estimating distances. A similar neurological syndrome has previously been described in older patients among the paraneoplasic syndromes associated with small-cell lung carcinoma and the presence of circulating anti-Hu antibodies, but never in the rare cancer-free patients with anti-Hu-associated chronic idiopathic intestinal pseudo-obstruction. The patient underwent a steroid treatment. No further episodes of functional intestinal obstruction were observed and, after an initial improvement, the neurological symptoms stabilized, leaving a permanent reduction in hearing function and an unsteady gait. The case shows that an idiopathic inflammatory insult of the myenteric plexus may precede (and perhaps lead to) central nervous system impairment in patients with anti-Hu associated chronic idiopathic intestinal pseudo-obstruction. PMID- 15756100 TI - Paroxysmal nocturnal hemoglobinuria and Budd-Chiari syndrome: therapeutic challenge with bone marrow transplantation, transjugular intrahepatic portosystemic shunt, and vena cava stent. AB - Stenosis of inferior vena cava can be a cause of or a consequence of Budd-Chiari syndrome (BCS). However, its occurrence after transjugular intrahepatic portosystemic shunt (TIPS) insertion has only been twice reported. We report the case of a 23-year-old man who presented BCS of the three suprahepatic veins. The cause of BCS was paroxysmal nocturnal hemoglobinuria, which was treated by bone marrow transplantation. A few months later, it was necessary to insert a TIPS because of refractory ascites and severe denutrition. However, refractory ascites was persistent and esophageal varices bleeding occurred. A TIPS desobstruction was needed, and during this angiography a stenosis of the suprahepatic vena cava was found and was treated with dilatation and stent. In a few weeks, the patient's clinical and nutritional state improved and, after 9 months hospitalization, the patient was discharged. One year later, this patient had no symptoms with an excellent permeability of TIPS and vena cava stent. This case report confirms that TIPS is a good treatment for BCS, but stenosis of inferior vena cava can occur after such a treatment. PMID- 15756101 TI - Promising treatment of autoimmune hepatitis with 6-thioguanine after adverse events on azathioprine. AB - The use of corticosteroids in autoimmune hepatitis is an established therapy. To avoid the possible serious side effects of corticosteroids, immunosuppression with azathioprine is often warranted. Azathioprine, a purine analogue, is frequently used to taper or replace corticosteroids. However, approximately 10% of the patients are intolerant to azathioprine. Alternative therapies using mycophenolate, tacrolimus, budesonide, cyclosporine and 6-mercaptopurine have been studied, with variable results. The use of 6-thioguanine, an agent more directly leading to the down-stream active metabolites of azathioprine (6 thioguanine nucleotides) in inflammatory bowel disease patients intolerant to azathioprine or 6-mercaptopurine showed conflicting results. We report three patients with autoimmune hepatitis who could not tolerate azathioprine but tolerated 6-thioguanine 0.3 milligram per kilogram daily well. All three patients improved clinically. Therapeutic drug monitoring was performed. The prospective evaluation of 6-thioguanine as a possible immunosuppressive drug in autoimmune hepatitis patients is warranted. PMID- 15756102 TI - Cirrhosis related chylous ascites successfully treated with TIPS. AB - We describe a patient with chylous ascites, who was extensively investigated for the cause. No malignant or lymphatic disease could be found, but a liver biopsy revealed liver cirrhosis. The chylous ascites was unsuccessfully treated with a sodium restriction diet, diuretics and a medium chain triglyceride diet. After the placement of a transjugular intrahepatic portosystemic shunt the ascites disappeared. PMID- 15756109 TI - Disaster management and the ABA Plan. PMID- 15756111 TI - The Pentagon attack of September 11, 2001: a burn center's experience. AB - On September 11, 2001, an airplane flown by terrorists crashed into the Pentagon, causing a mass casualty incident with 189 deaths and 106 persons treated for injuries in local hospitals. Nine burn victims and one victim with an inhalation injury only were transported to the burn center hospital. The Burn Center at Washington Hospital Center admitted and treated the acute burn patients while continuing its mission as the regional burn center for the Washington DC region. Eight of the nine burn patients survived. Lessons learned include 1) A large volume burn center hospital can absorb nine acute burns and maintain burn center and hospital operations, but the decision to keep or transfer burn patients must be tempered with the reality that several large burns can double or triple the work load for 2 to 3 months. 2) Transfer decisions should have high priority and be timely to ensure optimum care for the patients without need for movement of medical personnel from one burn center to another. 3) The reserve capacity of burn beds in the United States is limited, and the burn centers and the American Burn Association must continue to seek recognition and support from Congress and the federal agencies for optimal preparedness. PMID- 15756112 TI - A regional burn center's response to a disaster: September 11, 2001, and the days beyond. AB - This report reviews the response of a regional burn center to the disaster that occurred in New York City at the World Trade Center on September 11, 2001. In addition, it assesses that response in the context of other medical institutions in the region. There were facilities in the region that had 120 burn care beds; only two-thirds of the burn-injured patients who required hospital admission were admitted to designated burn centers, and only 28% of burn-injured victims initially were triaged to regional burn centers. The care rendered at this center was made possible by a "disaster-ready" facility and supplementation of personnel from the resources provided by The National Disaster Medical System. The patient outcomes at this center exceeded that as predicted by logistic regression analysis. PMID- 15756113 TI - The Bali burn disaster: implications and lessons learned. AB - In October 2002, a terrorist attack on a nightclub in Bali resulted in an explosion and fire, causing the deaths of more than 200 people, including 88 Australian citizens. After first aid and primary care, the injured were repatriated to Darwin for triage and continued treatment and were then disseminated to various burn units throughout Australia. At the Repatriation General Hospital Concord Sydney, we received 12 patients with burns and a variety of blast injuries. Their treatment was complicated by infection with multiresistant organisms that were previously unseen in our unit and the presence of complex shrapnel wounds. There were no deaths and, with two exceptions, all patients were discharged within 6 weeks. This incident had profound effects on our unit, particularly related to the management of high-velocity shrapnel injuries, serious ongoing septic complications, and the psychological effects on both patients and staff, all of which are detailed and discussed. PMID- 15756114 TI - The Pope Air Force Base aircraft crash and burn disaster. AB - This report describes the initial hospital and burn center management of a mass casualty incident resulting from an aircraft crash and fire. One hundred thirty soldiers were injured, including 10 immediate fatalities. Womack Army Medical Center at Fort Bragg, North Carolina, managed the casualties and began receiving patients 15 minutes after the crash. As a result of repetitive training that included at least two mass casualty drills each year, the triage area and emergency department were cleared of all patients within 2 hours. Fifty patients were transferred to burn centers, including 43 patients to the US Army Institute of Surgical Research. This constitutes the largest single mass casualty incident experienced in the 57-year history of the Institute. All patients of the US Army Institute of Surgical Research survived to hospital discharge, and 34 returned to duty 3 months after the crash. The scenario of an on-ground aircraft explosion and fire approximates what might be seen as a result of an aircraft hijacking, bombing, or intentional crash. Lessons learned from this incident have utility in the planning of future response to such disasters. PMID- 15756115 TI - The station nightclub fire. PMID- 15756117 TI - Burn support for Operation Iraqi Freedom and related operations, 2003 to 2004. AB - Thermal injury historically constitutes approximately 5% to 20% of conventional warfare casualties. This article reviews medical planning for burn care during war in Iraq and experience with burns during the war at the US Army Burn Center; aboard the USNS Comfort hospital ship; and at Combat Support Hospitals in Iraq and in Afghanistan. Two burn surgeons were deployed to the military hospital in Landstuhl, Germany, and to the Gulf Region to assist with triage and patient care. During March 2003 to May 2004, 109 burn casualties from the war have been hospitalized at the US Army Burn Center in San Antonio, Texas, and US Army Burn Flight Teams have moved 51 critically ill burn casualties to the Burn Center. Ten Iraqi burn patients underwent surgery and were hospitalized for up to 1 month aboard the Comfort, including six with massive wounds. Eighty-six burn casualties were hospitalized at the 28th Combat Support Hospital for up to 53 days. This experience highlights the importance of anticipating the burn care needs of both combatants and the local civilian population during war. PMID- 15756116 TI - Managing a combined burn trauma disaster in the post-9/11 world: lessons learned from the 2003 West Pharmaceutical plant explosion. AB - At 1:37 pm on January 29, 2003, an explosion occurred at the West Pharmaceutical chemical plant in Kinston, North Carolina. The explosion killed three people at the scene and resulted in more than 30 admissions to area hospitals. The disaster resulted in 10 critically ill burn patients, who were all intubated with inhalation injuries, many with combined burn and trauma injuries. All 10 critically injured patients were admitted to a tertiary care facility 100 miles away with both a Level I trauma center and a verified burn center. Ultimately, 7 of 10 patients survived (a mortality rate of 30%), and none were transferred to another trauma or burn center. This article analyzes the unique challenges that combined burn and trauma patients present during a disaster, critically examines the response to this disaster, describes lessons learned, and presents recommendations that may improve the response to such disasters in the future. PMID- 15756118 TI - Combat burn life support: a military burn-education program. AB - The Advanced Burn Life Support Course has been used to train military physicians and nurses for more than 16 years. Although it useful for teaching the fundamentals of burn care, the course is designed for a civilian audience, covers only the first 24 hours of burn care, and presumes the availability of a burn center for patient transfer. In preparation for hostilities in Iraq, we developed several add-on modules to the standard Advanced Burn Life Support course to meet specific needs of military audiences. These modules cover the treatment of white phosphorus burns; the treatment of mustard gas exposure; the long-range aeromedical transfer of burn patients; the management of burn patients beyond the first 24 hours; and the delivery of burn care in austere environments. These add on modules are termed Combat Burn Life Support. Between January 22, 2003, and May 12, 2003, Advanced Burn Life Support and/or Combat Burn Life Support courses were provided to a total of 1035 military health care providers in the United States, Germany, and the Middle East. Student feedback was largely positive and is being used for further course refinement. The Combat Burn Life Support Course is designed to augment, rather than replace, the Advanced Burn Life Support Course. Although intended for a military audience, the course material is equally applicable to civilian terrorist or mass casualty situations. PMID- 15756119 TI - Nursing's role in the planning, preparation, and response to burn disaster or mass casualty events. PMID- 15756120 TI - Burn specialty teams. AB - Natural disasters have always been a threat. human-caused disasters, especially terrorist acts, are increasing in frequency. Burn centers and providers have an important contribution to make in caring for those injured in these incidents. The most effective way to make a contribution is to act in cooperation with the Federal Disaster Response, which is organized by the Department of Homeland Security and the Federal Emergency Management Agency. It appears that this can be most effectively accomplished through participation in the Burn Specialty Team Program, which has been developed to rapidly augment emergency medical teams with burn expertise. PMID- 15756121 TI - Tracking the daily availability of burn beds for national emergencies. AB - Medical planning for Operation Iraqi Freedom included predictive models of expected number of burn casualties. In all but the best-case scenario, casualty estimates exceeded the capacity of the only Department of Defense burn center. Examination of existing federal-civilian disaster plans for military hospital augmentation revealed that bed availability data were neither timely nor accurate. Recognizing the need for accurate knowledge of burn bed availability, the Department of Defense requested assistance from the American Burn Association (ABA). Directors of burn centers in the United States were queried for interest in participation in a mass casualty plan to provide overflow burn bed capacity. A list of 70 participating burn centers was devised based upon proximity to planned military embarkation points. A computer tracking program was developed. Daily automated e-mail messages requesting bed status were sent to burn center directors at 6 am Central time with responses requested before 11 am. The collated list of national overflow burn bed capacity was e-mailed each day to the ABA Central Office and to federal and military agencies involved with burn patient triage and transportation. Once automated, this task required only 1-2 hours a day. Available burn-bed lists were generated daily between March 17 and May 2, 2003 and then every other day until May 9, 2003. A total of 2151 responses were received (mean, 43 burn centers per day). A system to track daily nationwide burn bed availability was successfully implemented. Although intended for military conflict, this system is equally applicable to civilian mass casualty situations. We advocate adoption of this or a similar bed tracking system by the ABA for use during burn mass casualty incidents. PMID- 15756122 TI - Plenary Session II: American Burn Association Disaster Readiness Plan. PMID- 15756123 TI - Autonomic disorders and the eye. PMID- 15756124 TI - Segmental facial anhidrosis and tonic pupils with preserved deep tendon reflexes: a novel autonomic neuropathy. AB - A 31-year-old woman had exertional right-sided hemifacial flushing and sweating. Examination demonstrated slightly dilated pupils with absent constriction to light and a tonic near response and redilatation, features consistent with Adie syndrome. Neurological examination was otherwise normal, including preservation of deep tendon reflexes. Magnetic resonance imaging of brain and spine were normal. The combination of unilateral loss of sudomotor and vasomotor activity without loss of ocular sympathetic innervation fulfills the diagnosis of Harlequin syndrome. The combination of Harlequin and Adie syndromes has been called Ross syndrome, but the preservation of deep tendon reflexes precludes a diagnosis of Ross syndrome in our patient. This previously undescribed variant adds further complexity to the spectrum of autonomic neuropathies. PMID- 15756125 TI - Nonarteritic ischemic optic neuropathy developing soon after use of sildenafil (viagra): a report of seven new cases. AB - Seven patients, aged between 50 and 69 years, had typical features of nonarteritic anterior ischemic optic neuropathy (NAION) within 36 hours after ingestion of sildenafil citrate (Viagra) for erectile dysfunction. Six patients had vision loss within 24 hours after use of the agent. Final visual acuity in the affected eye ranged from 20/20 to light perception. Both eyes were affected in one individual. All affected individuals had pre-existing hypertension, diabetes, elevated cholesterol, or hyperlipidemia. Seven similar cases have been previously reported. Sildenafil may provoke NAION in individuals with an arteriosclerotic risk profile. PMID- 15756126 TI - Predictors of recurrent ischemic optic neuropathy in giant cell arteritis. AB - BACKGROUND: The competing interests of preventing recurrent ischemic optic neuropathy (ION) and minimizing medication side effects make corticosteroid dose reduction in giant cell arteritis (GCA) a difficult problem. The authors sought to determine whether any factors were predictive of recurrent ION. METHODS: Retrospective review of the records of 100 consecutive patients with biopsy proven giant cell arteritis diagnosed in two Australian hospitals between 1988 and 1998. Among 67 patients who met inclusion criteria for ION in GCA, seven patients had recurrent ION. We compared the seven patients with recurrent ION to the 60 patients with nonrecurrent ION in terms of age, gender, mode of corticosteroid delivery, initial visual acuity in the affected eye, prevalence of bilateral ION, initial erythrocyte sedimentation rate (ESR) level, and rate of corticosteroid dose reduction. In the recurrent ION group, we documented the timing of the recurrence in relation to corticosteroid dose, elevation in acute phase reactants, and relapse of systemic symptoms. RESULTS: We found recurrent ION in GCA in 10% of our cohort, higher than has been previously reported. Recurrences, all of which were ipsilateral, occurred from 3 to 36 months (median 8 months) after the initial ION. None of the clinical indicators the authors examined differed between the two groups. Six of seven patients with recurrent ION had elevations in ESR or C-reactive protein or a new headache at the time of ION recurrence, but in only one of these patients were these features recognized as preceding the recurrent ION. One patient had neither an elevation in acute phase reactants nor a relapse in systemic symptoms of GCA at the time of ION recurrence. CONCLUSIONS: Recurrent ION in GCA is difficult to predict. Although elevated acute phase reactants or new systemic symptoms consistent with GCA were present in six (83%) of our patients with ION recurrence, in only one patient (17%) did these events occur with enough lead time to allow caregivers to act preemptively. Thus, even very close monitoring of GCA patients with ION may not predict ION recurrence. PMID- 15756127 TI - Optic neuropathy associated with linezolid treatment. AB - Two patients undergoing long-term treatment with linezolid for pneumonia had decreased visual acuity, dyschromatopsia, and cecocentral scotomas characteristic of toxic optic neuropathy. Visual function slowly recovered 3 to 4 months after discontinuation of the antibiotic treatment in both patients. Toxic optic neuropathy may occur from linezolid. Early withdrawal of the antibiotic may be associated with visual recovery. PMID- 15756128 TI - A comparison of risk factors for postoperative and spontaneous nonarteritic anterior ischemic optic neuropathy. AB - BACKGROUND: : Whether postoperative non-arteritic ischemic optic neuropathy (NAION) is caused by surgery is unsettled. To provide further evidence on this issue, it is useful to compare the characteristics of patients who develop NAION following intraocular surgery to those who develop NAION spontaneously. METHODS: : In a retrospective review of patients diagnosed with NAION between January 1, 1993 and December 31, 1999, 12 cases of NAION in 11 subjects were identified as occurring within 30 days of cataract extraction or intraocular lens exchange (postoperative NAION group). Using Fisher exact test, the prevalence of NAION risk factors (hypertension, diabetes mellitus, hypercholesterolemia, smoking, small cup-to-disc ratios) was compared with that of a similarly aged control group of 37 subjects diagnosed with spontaneous NAION (spontaneous NAION group). RESULTS: : Patients with postoperative NAION had a lower prevalence of hypertension than did those with spontaneous NAION (27% versus 68%, P = 0.034) and a lower prevalence of cup-to-disc ratios of less than or equal to 0.2 (55% vs. 94%, P = 0.006). The prevalence of elevated cholesterol, diabetes mellitus, and tobacco use was similar in the two groups. CONCLUSIONS: : The prevalence of hypertension and low cup-to-disc ratios is significantly lower in subjects with NAION following lens-related intraocular surgery than in those with spontaneous NAION, indicating that risk factors for NAION in these settings may be different. PMID- 15756129 TI - Bilateral optic neuritis in wegener granulomatosis. AB - A 32-year-old patient with recurrent sinusitis had severe visual loss from optic neuropathy. Imaging revealed severe bone destruction and soft tissue densities of the paranasal sinuses and enhancement of the dura of the frontal sinuses, optic canals, and superior orbital fissures bilaterally. Endoscopic sinusectomy with biopsy showed granulomatous vasculitis compatible with Wegener granulomatosis (WG). The patient was treated with intravenous and oral corticosteroids and oral cyclophosphamide that led to rapid and dramatic visual recovery. This case draws attention to the fact that optic neuritis may be an early inflammatory manifestation of WG and that rapid diagnosis and aggressive anti-inflammatory treatment is critical before inflammation of arteries leads to infarction and irreversible visual loss. PMID- 15756130 TI - Bilateral simultaneous central retinal artery occlusions in wegener granulomatosis. AB - A 46-year old woman developed simultaneous central retinal artery occlusions (CRAOs) in Wegener granulomatosis (WG). She had presented six years earlier with xerostomia, skin rash, and arthralgias and received a diagnosis of Sjogren syndrome. Anti-neutrophilic cytoplasmic antibody (ANCA) was negative. Months prior to the CRAOs, she had developed hearing loss, proptosis, and scleritis that were not responsive to prednisone 50 mg/d. The CRAOs occurred while she was being treated at this dose level. ANCA was now positive. This is the 12th case of CRAO in WG and the 6th case of bilateral CRAOs reported in the English literature. It emphasizes that serious irreversible visual complications may occur even when the patient is being treated with substantial corticosteroid doses. PMID- 15756131 TI - The elusive nature of primary intraocular lymphoma. AB - A 58-year-old woman with an initial diagnosis of multiple evanescent white dot syndrome OU experienced deteriorating vision despite corticosteroid treatment. Reevaluation disclosed retinal and subretinal infiltrates and pigmentary alterations, prompting a suspicion of primary intraocular lymphoma (PIOL). Diagnostic vitrectomy yielded inconclusive cytology, but flow cytometry demonstrated small monoclonal B cells less suggestive of PIOL than of small lymphocytic lymphoma originating outside the eye or central nervous system. Brain magnetic resonance imaging, chest/abdomen/pelvis computed tomography, lumbar puncture, and laboratory studies failed to disclose lymphoma elsewhere. There was insufficient evidence to recommend radiation therapy. Vision deteriorated rapidly, prompting a diagnostic retinal biopsy and aspiration of the subretinal infiltrate, revealing unequivocal evidence of PIOL. After 40 Gy orbital x irradiation, visual function improved dramatically. This case emphasizes the unusual ocular manifestations of PIOL and the difficulty of obtaining a definitive diagnosis by sampling vitreous, particularly after corticosteroid treatment. In such cases, subretinal aspiration or retinal biopsy may be necessary. Timely diagnosis is critical because treatment can reverse visual loss if it is not severe. PMID- 15756132 TI - Multiple bilateral extraocular muscle metastases as the initial manifestation of breast cancer. AB - A 75-year-old woman had gradually progressive binocular diplopia and 2 months later had breast cancer diagnosed. Examination showed bilateral external ophthalmoplegia with lid retraction and lag and no pupillary abnormalities. Orbital magnetic resonance imaging showed enlargement and enhancement of all extraocular muscles bilaterally. A right orbital biopsy was consistent with metastatic breast carcinoma. Positron emission tomography, bone scan, and computed tomography of the chest, abdomen, and pelvis failed to disclose other evidence of breast cancer metastases. It is unusual to encounter metastatic breast cancer affecting every extraocular muscle before the diagnosis of the primary carcinoma. PMID- 15756133 TI - A mechanical theory to account for bitemporal hemianopia from chiasmal compression. AB - The association between bitemporal hemianopia and chiasmal compression is well recognized. The majority of chiasmal syndromes are caused by extrinsic compression from pituitary tumors, suprasellar meningiomas, craniopharyngiomas, and aneurysms. However, it is not clear why compressive lesions of the chiasm show a predilection for damage to nasal fibers with bitemporal hemianopia. Few experimental attempts at elucidating these mysteries have been reported and none has provided an adequate explanation. The authors postulate that the susceptibility of nasal fibers to preferential damage is explained by structural collapse theories as applied to crossing and noncrossing cylinders. By constructing a simplified mathematical model, the authors demonstrate that nasal fibers are subject to relatively greater pressures for any given external compressive force acting on the chiasm. PMID- 15756134 TI - Otto lowenstein, pioneer pupillographer. AB - Otto Lowenstein, a pioneer in the study of pupil function, began his professional life as an academic neuropsychiatrist at the University of Bonn with an interest in experimental psychology. From his teacher Alexander Westphal, he developed a fascination with the pupil. He invented ingenious recording devices and took motion pictures of the pupils, graphing their movements. Forced to flee Nazi persecution in 1933, he took temporary refuge in Switzerland and eventually sacrificed a flourishing career in Europe to escape to New York. During the next 25 years, he collaborated with Irene Loewenfeld on experiments and publications related to the clinical use of pupillary signs and introduced pupillography to American neuro-ophthalmology. PMID- 15756135 TI - Second Congress of the Asian Neuro-Ophthalmology Society, Nagoya, Japan, October 15-16, 2004. PMID- 15756136 TI - Bilateral optic neuritis after dengue viral infection. PMID- 15756137 TI - One-and-a-half syndrome with facial diplegia: the 15 1/2 syndrome? PMID- 15756138 TI - Unilateral involuntary eyelid closure induced by diplopia that did not remit with contact lens occlusion. PMID- 15756139 TI - Progressive external ophthalmoplegia with hereditary sensory neuropathy: a rare association. PMID- 15756140 TI - See-saw nystagmus as the presenting sign in multiple sclerosis. PMID- 15756142 TI - Etiological factor of hepatocellular carcinoma in Italy. AB - Cirrhosis from any cause is present in the majority (93.1%) of hepatocellular carcinoma (HCC) cases in Italy, it seems to be the common pathway by which several risk factors extent their carcinogenic effect. The mortality rate of HCC in Italy has progressively increased during the period 1969-1994, reflecting the rising number of persons living with cirrhosis as consequence of the remarkable advances in medical management of such patients. Most HCC develops in cirrhosis caused by known and preventable risk factors (hepatitis B virus, HBV, hepatitis C virus, HCV, alcohol and possibly non-alcoholic steatohepatitis, NASH). Unlike alcohol and NASH, HBV and HCV chronic infections act as a risk factors for HCC both because they induce cirrhosis and because they increase the risk in patients with cirrhosis. Moreover, case-control and prospective studies have shown a synergistic effect on HCC risk, when both viral infections occur. Currently, HCV infection is detected in the majority (76.4%) of HCC cases in Italy, reflecting the large cohort of subjects infected with this virus via the iatrogenic route during the 1950s and 1960s. The low rate of HCV infection in the younger Italian generations may generate a downward trend in the HCC mortality rate in the years to come. PMID- 15756143 TI - Risk factors for the acquisition of hepatitis C. AB - About 170 million people are estimated to be chronically infected with hepatitis C worldwide. Despite intense research efforts and considerable improvement of antiviral therapy sustained virological response can only be achieved in a minority of patients due to viral non-response or relapse, contraindications against therapy and high costs. Chronic hepatitis C is the leading cause for end stage liver disease, hepatocellular carcinoma and liver transplantation in developed countries. To limit the incidence of new infections knowledge about potential transmission pathways and risk factors is crucial. This article aims to summarize recent advances in understanding transmission pathways and outlines future prevention strategies. PMID- 15756144 TI - Natural history of hepatitis C virus infection: from chronic hepatitis to cirrhosis, to hepatocellular carcinoma. AB - Hepatitis C is a heterogeneous disease and is responsible for considerable mortality and morbidity. The hepatitis C virus (HCV) infects nearly 170 million people world-wide. More than 80% of infected individuals develop chronic infection; the remaining 10-20% develop spontaneous clearance with natural immunity. Acute hepatitis is icteric in only 20% of patients and is rarely severe. The majority of patients who develop chronic HCV infection are asymptomatic; but 60-80% develop chronic hepatitis as indicated by elevated alanine aminotransferase (ALT), around 30% maintain persistently normal ALT levels despite having detectable HCV-RNA in serum. One-third of chronically infected patients develop progressive liver injury, fibrosis and cirrhosis over a period of 20-30 years. The relationship between virus load, HCV genotype, quasi species variability and progression of liver disease is controversial. Acquired infection after age 40, male sex, excessive alcohol-consumption, hepatitis B virus (HBV) or HIV co-infection, steatosis, and immunosuppressed state have been identified as co-factors associated with progression of fibrosis and development of cirrhosis. In patients with cirrhosis, the incidence of hepatocellular carcinoma is 2-5% per year. At present, HCV-related end-stage cirrhosis is the first cause of liver transplantation. PMID- 15756145 TI - Non-responders to previous treatment for hepatitis C. AB - The main principle in treating chronic hepatitis C is the prevention of serious liver complications. Because curing hepatitis C virus infection has been demonstrated to prevent progression of liver disease and even promote regression of fibrosis, it remains the primary goal of treatment. However, nearly half of patients are not cured with our best treatment. Patients who failed older therapies should be treated with peginterferon and ribavirin, but those with advanced fibrosis or African heritage will have very low rates of response. Non responders to peginterferon and ribavirin present a special challenge. If there were problems related to dosing, adherence, or access during treatment, then one can consider re-treating with the same regimen if the problems can be corrected. Otherwise, non-responders with early-stage fibrosis can observe without further treatment until newer drugs are available. Those with advanced fibrosis should consider low-dose peginterferon maintenance treatment or participation in an experimental trial. Experimental approaches include intensification of existing therapies, combination of new agents with existing drugs, long-term virus suppression, inhibition of liver fibrogenesis, and inhibition of hepatitis C RNA or hepatitis C viral enzymes. PMID- 15756146 TI - Autoimmune manifestations during interferon therapy in patients with chronic hepatitis C: the hepatologist's view. AB - In face of numerous benefits induced by therapy based on interferon (IFN) associated with ribavirin for the treatment of chronic hepatitis C, there is an increasing concern regarding its tolerance, which can, in some cases, reduce the quality of life as well as compliance of patients. Among the less common side effects, there are the autoimmune ones which can be globally divided into appearance or increase in titres of auto-antibodies and/or manifestation of overt autoimmune pathologies. Whereas the former may concern more than 50% of treated subjects, the latter is reported in only 1-2% of patients under therapy. Thyroid dysfunction represents the well-studied autoimmune disorder. The presence of pre existing anti-thyroid antibodies and being of female sex, constitute relevant risk factors for the development of a disease involving this gland. Often the treatment of thyropathy must be continuous in spite of IFN discontinuation because the disturbance usually does not abate with stopping antiviral therapy. Some observations have pointed out to the fact that IFN can lead to the development of insulin-dependent diabetes mellitus. Sometimes, during, as well as after IFN treatment, the appearance of anti-islet cell antibodies has been shown, but its interrelationship with the development of disease is uncertain. While being treated with IFN for chronic hepatitis C, the finding of non-organ specific antibodies at baseline can increase the likelihood of the development of autoimmune hepatitis. However, their presence does not constitute an absolute contraindication to the treatment, except in case of high titre. Other disorders, such as a lupus erythematosus-like syndrome, haemolytic anaemia, and immune mediated thrombocytopenia have been reported. In conclusion, although the presence of auto-antibodies is considered to be an epiphenomenon without pathogenic significance in most patients suffering from chronic hepatitis C, it poses a problem when they need to be treated with IFN. This antiviral drug can induce or exacerbate a multitude of autoimmune-related disorders, however, clinically overt immune-mediated diseases are rare and affect a subset of subjects who have an underlying autoimmune diathesis. PMID- 15756147 TI - Natural history of hepatitis B. AB - The natural course of hepatitis B virus (HBV) chronic infection is variable, ranging from an inactive HBsAg carrier state to a more or less progressive chronic hepatitis, potentially evolving to cirrhosis and hepatocellular carcinoma (HCC). Chronic hepatitis may present as typical HBeAg-positive chronic hepatitis B or HBeAg-negative chronic hepatitis B. HBeAg-positive chronic hepatitis is due to wild type HBV; it represents the early phase of chronic HBV infection. HBeAg negative chronic hepatitis is due to a naturally occurring HBV variant with mutations in the precore or/and basic core promoter regions of the genome; it represents a late phase of chronic HBV infection. The latter form of the disease has been recognized as increasing in many countries within the last decade and it represents the majority of cases in many countries. HBeAg-negative chronic hepatitis B is generally associated with a more severe liver disease with a very low rate of spontaneous disease remission and a low sustained response rate to antiviral therapy. Longitudinal studies of patients with chronic hepatitis B indicate that, after diagnosis, the 5-year cumulative incidence of developing cirrhosis ranges from 8-20%. Morbidity and mortality in chronic hepatitis B are linked to evolution to cirrhosis or HCC. The 5-year cumulative incidence of hepatic decompensation is approximately 20%. The 5-year probability of survival is approximately 80-86% in patients with compensated cirrhosis. Patients with decompensated cirrhosis have a poor prognosis (14-35% probability of survival at 5 years). HBV-related end-stage liver disease or HCC are responsible for at least 500,000 deaths per year. PMID- 15756148 TI - Current and future treatment of chronic hepatitis B. AB - Hepatitis B virus (HBV) infection is an ongoing worldwide pandemic. In general, the mode of transmission is different according to its prevalence in different areas: mostly perinatal in highly prevalent zones and predominantly sexual and/or parenteral in low prevalence areas. The variation in presentations of chronic hepatitis B (CHB) make its management complex. Aminotransferase levels, viral load, histologic changes, age of the patient, viral mutations (e.g. hepatitis B e antigen negative) and pregnancy are all factors that impact on treatment decisions. The ideal drug that will eradicate the HBV has yet to be developed. This review focuses on the currently available medications for CHB: alpha interferon, lamivudine, adefovir, as well as new drugs that have proven to be active against HBV in clinical trials and are closer to licensure; pegylated interferon tenofovir, entecavir, emtricitabine, telbivudine and clevudine. The antiviral properties and the advantages and disadvantages in HBeAg-positive and negative patients are discussed. Combinations of different medications currently under study were not included in this review. A suggested algorithm, developed from recent literature, may serve as a practical guide on tailoring drug selection based on viral load, aminotransferases, hepatitis B e antigen status and histological findings. Finally, practical management questions are raised. PMID- 15756149 TI - Genetic variants of hepatitis B virus and their clinical relevance. AB - Infection with hepatitis B virus (HBV) leads to a wide spectrum of clinical presentations ranging from an asymptomatic carrier state to self-limited acute or fulminant hepatitis to chronic hepatitis with progression to cirrhosis and hepatocellular carcinoma (HCC). Infection with HBV is one of the most common viral diseases affecting man. Both viral factors as well as the host immune response have been implicated in the pathogenesis and clinical outcome of HBV infection. Evidence has been accumulating that HBV mutants are associated with certain clinical disease manifestations, may affect the natural course of the infection and confer resistance to antivirals. Naturally occurring mutations have been identified in the structural and non-structural genes as well as regulatory elements of the virus. The best characterized mutants comprise the pre-core (pre C) stop codon mutation resulting in a loss of hepatitis B e antigen (HBeAg), defined clusters of mutations in the core promotor resulting in enhanced viral replication and mutations in the hepatitis B core and surface antigens (HBcAg and HBsAg) altering the antigenicity of the virus. More recently, several mutations in the reverse transcriptase/polymerase gene have been identified conferring resistance to antivirals used for the treatment of chronic hepatitis B. In this review, we will focus on the biological phenotype of HBV genetic variants and discuss their clinical relevance for the pathogenesis of HBV-induced liver disease. PMID- 15756150 TI - An update in liver transplantation in patients with hepatitis B and hepatitis C. AB - Chronic hepatitis C virus (HCV) infection is an epidemic that currently represents the number one indication for liver transplantation (LTx). Hepatitis B virus (HBV) infection is associated with better outcomes following LTx since the advent of hepatitis B immune globulin and lamivudine. The impact of HCV and HBV in LTx is well known. Therapeutic interventions, however, are less standardized and often depend upon institutional protocol. This review article will provide a comprehensive review of the literature and address many issues and complications with transplantation in patients suffering from chronic liver disease as a result of HCV or HBV. PMID- 15756151 TI - Epidemiology of hepatitis C virus infection. AB - Although a lot of novel information and data on the epidemiology of hepatitis C virus (HCV) infection are available worldwide, the majority of these information are often fragmentary and sometimes contradictory. This review tries to highlight all the data available on the prevalence (i.e. the number of cases present in a known population), the risk factors, the natural history and the incidence (i.e. the number of new cases that occur every year) of HCV infection in the world, and particularly in Italy. PMID- 15756152 TI - When plans backfire: telling the difference between the pain physician and the palliative care physician. PMID- 15756154 TI - Perioperative analgesia in pulmonary surgery. AB - AIM: In pulmonary surgery many studies have shown how pain associated with residual doses of anaesthetic drugs can cause a decrease in pulmonary volumes and depression of the cough reflex. Both these phenomena are responsible for a rising incidence of postoperative mortality and morbidity. The most widely used postoperative analgesic techniques are continuous systemic analgesia and epidural analgesia. The aim of this study is to verify the advantages and the disadvantages of both analgesic techniques. METHODS: Fifty patients, undergoing pulmonary surgery, were recruited and divided, after randomization into 2 groups. Patients included in A group received an epidural administration of naropina 0.2%+fentanyl 4 microg/mL by elastomeric pump (rate 5 ml/h). Patients included in B group received an ev continuous infusion of tramadol 600 mg+ketorolac trometamina 120 mg+ranitidina 200 mg+ondansetron 16 mg by elastomeric pump (rate 5 ml/h for 48 hours). RESULTS: Both groups showed good analgesic effects. Pain rest relief was between 3 and 1.7 points in group B and between 2.5 and 0.4 points in group A. Incident pain was 4.8 at awakening time and it decreased to 4 after 48 hours in group B while in group A it was from 3.2 to 1.8 in the same period of B group. CONCLUSIONS: Our data show that both analgesic techniques are able to guarantee a good rest pain relief after thoracotomy. Epidural analgesia showed more efficacy as far as incident pain relief but it was more difficult to realise and it showed less acceptance by patients. PMID- 15756153 TI - Peripheral and central mechanisms of orofacial pain and their clinical correlates. AB - This paper reviews recent research that has provided important insights into the peripheral processes by which noxious stimuli activate or modulate nociceptive afferent inputs into the brainstem, that has identified the critical neural elements and pathways in the brainstem and higher levels of the trigeminal (V) somatosensory system that receive and transmit nociceptive inputs from orofacial tissues, and that has clarified some of the mechanisms involved in the modulation and plasticity of nociceptive transmission. The paper also outlines some of the clinical correlates of these research advances. PMID- 15756155 TI - Desflurane and sevoflurane in elderly patients during general anesthesia: a double blind comparison. AB - AIM: To investigate pulmonary wash-out of sevoflurane and desflurane and the quality of recovery from anesthesia in elderly patients. METHODS: Thirty-six patients aged >65 years, ASA II, were assigned in a double blind fashion to either desflurane (n=18) or sevoflurane (n=18) anesthesia. All received propofol 2 mg/kg and remifentanil 0.2 microg/kg/min for induction and 0.6 mg/kg of rocuronium. When the trachea was intubated volatile anaesthetic was administered. All data were recorded 1, 3, 5, 15, 30 min after intubation and then every 15 min. All data were recorded 1, 2, 3, 4, 5 min after suspension of all agents. Once extubated simple orders and questions were given every minute, times of appropriate response were noted. The patients were then transferred to the recovery room, until discharge to the floor. Postoperative pain control was obtained by a continuous iv infusion of ketorolac 60 mg and tramadol 100 mg. The latter was incremented by supplemental boluses of 50 mg according to patient needs (VAS <4) up to a maximum of 300 mg/24h. RESULTS: The F(A)/F(A0) ratio was lower in the desflurane group after halogenated agent suspension (p= or <0.05). Desflurane proved to have a faster wash out curve with respect to sevoflurane. Early recovery, as indicated by the time necessary to appropriately answer simple questions after the discontinuation of anesthetics, showed a significant advantage for desflurane (p= or <0.05). VAS was higher in the desflurane group as well as the needs for postoperative analgesia. CONCLUSIONS: Patients receiving desflurane reported faster recovery from anesthesia but an earlier and more intense perception of pain after surgery. PMID- 15756156 TI - Comparison of two protocols of conscious analgosedation in video-assisted talc pleurodesis. AB - AIM: Video-assisted thoracoscopy surgery (VATS) is classically performed using general anesthesia with a double-lumen endotracheal tube to allow collapse of the operated lung. However, according to our opinion, the risks of general anesthesia with one-lung ventilation could be accepted when major thoracic operation is planned, but it should be avoided or kept at minimum when performing less invasive procedures such as video-assisted talc pleurodesis. In this paper, 2 different protocols are described in order to demonstrate the effectiveness and safety of Monitored Anesthesia Care (MAC) for performing VATS talc pleurodesis. METHODS: We studied 65 neoplastic patients ASA III-IV who underwent video assisted pleurodesis with talc nebulization. They were randomized into 2 homogenous groups: Group 1, received midazolam (0.015-0.030 mg/kg) and sufentanil (0.15-0.20 microg/kg); Group 2, received midazolam (0.15-0.2 mg/kg) and continuous remifentanil infusion 0.5-1 microg/kg/min in the first minute, followed by 0.05 microg/kg/min. Local anesthesia with 0.2% ropivacaine was employed before the thoracoscopic trocar was inserted in both groups. Intraoperative standard monitoring required ECG, heart rate, non-invasive blood pressure, transcutaneous carbon dioxide and oxygen saturation. Consciousness status has been evaluated by Ramsey scale, while pain intensity by VAS scale. RESULTS: No statistically significant fluctuations were observed for mean arterial pressure, heart rate, SpO(2), tCO(2), VAS and Ramsey score. No postoperative complications and hospital mortality occurred. CONCLUSIONS: Both the protocols offer an efficient control of analgosedation with a minimum incidence of intraoperative and postoperative side effects. Anyway, the titration of the drug is very important. PMID- 15756157 TI - Post-traumatic stress disorder-related symptoms after intensive care. AB - AIM: To determine the incidence of Post Traumatic Stress Disorder (PTSD) related symptoms in a population of intensive care unit (ICU) admitted patients and the relationship between PTSD-related symptoms and memories of ICU. METHODS: Adults consecutively admitted to an ICU of a University hospital during 1 year, who stayed in the ICU at least 3 days, were prospectively studied. A questionnaire (ICU memory tool) was administered to 84 patients 1 week after ICU discharge and to 63 of them after 3 months. Past medical history and clinical variables present during ICU stay were collected. RESULTS: At the 1st interview, 5 patients (5.9%) did not remember to have been in ICU. Of the remaining 79 patients (males 59.5%, median age 69 years, SAPS II 34, APACHE II 14 and ICU stay 5 days), 4 reported intrusive memories and none panic attacks. The Impact of Events Scale (IES), available in 3 of them, scored in medium/high levels. Only the median number of factual memories reported by the patients with and without intrusive memories was significantly different (4 interquartile range 2-5 vs 8 interquartile range 6-10; p=0.002). The patients with intrusive memories at the 1st interview did not report them at the 2nd interview. Two patients not having panic or intrusive memories at the 1st interview reported PTSD-related symptoms after 3 months. CONCLUSIONS: In a general ICU population, few patients (5%) have PTSD-related symptoms and those who present those symptoms report less factual memories of ICU stay. PMID- 15756158 TI - Development of aerobic power in pubescent male soccer players related to hematocrit, hemoglobin and maturation. A longitudinal study. AB - AIM: More than ever young athletes are training intensively and compete at high levels. As adolescent athletes are training in a period associated with many changes in growth and maturation longitudinal studies are important to elucidate the development in physical parameters. The aim of this study was to evaluate the development in aerobic power in elite as well as non-elite soccer players during the years of puberty and to relate it to maturation as well as hemoglobin and hematocrit. METHODS: Forty-nine young male soccer players at elite and non-elite levels (10.5-13 y) were recruited as subjects. All subjects were tested 4 times over a period of 3.5 years. Anthropometrical parameters including skinfold were measured. VO2max was assessed during treadmill run, testicular volume and testosterone levels were used to estimate maturation, and hemoglobin (Hb) as well as hematocrit (Htc) was measured from blood samples. RESULTS: A longitudinal development in VO2max (l/min) was found for all players together. The elite players have higher values and steeper increase compared to non elite-players. This difference was still present when VO2max was adjusted for body weight (ml/kg/min). CONCLUSIONS: The general development in VO2max was significantly correlated to the development in Hb and Htc as well as maturation, but the difference between elite and non-elite players could not be explained from differences in these variables. As elite players often are selected at a very young age it is possible that training effect is present but it cannot be excluded that genetic factors may play an important role. PMID- 15756159 TI - The effects of rest interval and resistance training on quadriceps femoris muscle. Part II: EMG and perceived exertion. AB - AIM: The purpose of this study was to examine the effects of rest interval on quadriceps femoris muscle activation and perceived exertion, during short-term resistance training. METHODS: Vastus medialis (VM) and vastus lateralis (VL) muscle electromyograms (EMG) were assessed in 15 males during a sustained 80% maximal voluntary contraction (MVC). During the pre-training evaluation, the absolute value of the 80% MVC (N.m) and contraction duration (s) was performed at 2, 4, and 6 weeks during the training period. Perceived exertion was measured via the Borg category-ratio scale every 5 s during the 80% MVC. Subjects were randomly assigned to 3 groups: group 1 received a 40 s rest interval in between exercise sets, group 2 received a rest period of 160 s, and the control group did not participate in training. Groups 1 and 2 performed isokinetic knee extensions at 180 deg.s(-1) 2 days per week for 6 weeks. RESULTS: The results demonstrated a significant decrease in VM EMG within the initial portion of the 80% MVC across the training period in the short rest interval group. The long rest interval and control groups showed no significant changes in VM EMG during 1st part of the contraction across the training period, whereas the control group exhibited a significant reduction in VL EMG across weeks 4 to 6. VL EMG increased during the 80% MVC in the control group across the training period. VM EMG increased during the sustained contractions in the long rest interval and control groups across the training period. The perceived exertion response was lower in the 1st part of the 80% MVC in the short and long rest interval groups, but not in the control group, across the training period. The results also showed a significant decrease in perceived exertion at the end of the sustained contraction in the short rest interval group, but not in the long rest interval group or the control group. CONCLUSIONS: The findings from this study suggest that the application of relatively short rest intervals in between sets of resistance exercise induced a greater neuromuscular response of the VM muscle during short-term training. PMID- 15756161 TI - Surface electromyography of nine shoulder muscles in two iron cross conditions in gymnastics. AB - AIM: The rules of gymnastics impose an element of static strength such as an iron cross (IC) on the rings. For IC training, coaches use a custom made device -- the herdos -- to simulate the conditions of competition. The purpose was to compare muscle activity and coordination during IC performed both on the rings and using herdos. Secondly, we tried to determine whether herdos usage induced functional adaptations of the shoulder muscles. METHODS: Six male gymnasts performed 10 IC in each condition. Surface electromyogram of muscles pectoralis major, latissimus dorsi, teres major, infraspinatus, rhomboideus, trapezius, serratus anterior, biceps brachii, and triceps brachii in the right shoulder were analysed using root-mean-square (RMS), and muscle part's in each condition. Muscle part represents the contribution of each of the 9 shoulder muscles studied in each condition. Total muscle activity (SUM) was also used to compare the 2 conditions. RESULTS: Except for the muscle teres major, the RMS decreases (p<0.05) when using the herdos. The SUM also decreases (p<0.05) when using this device. The muscle parts indicate that the contribution of the muscle latissimus dorsi decreases (p<0.05) when using the herdos. These results suggest that the herdos modified shoulder coordination. But their usage does not seem to induce any functional adaptations of these muscles. CONCLUSIONS: The herdos do not seem to provide a valid method to reproduce the same shoulder coordination as on the rings. Therefore IC training with this special device could be called in question. PMID- 15756160 TI - The discriminative power of the Interval Shuttle Run Test and the Maximal Multistage Shuttle Run Test for playing level of soccer. AB - AIM: The purpose of this study was to examine the discriminative power of the recently developed Interval Shuttle Run Test (ISRT) and the widely used Maximal Multistage 20 m Shuttle Run Test (MMSRT) for soccer players at different levels of competition. The main difference between the tests is that the exercise mode of the ISRT is not continuous but intermittent. Instead of 60 s of running per stage in the MMSRT, 30 s of running are alternated with 15 s of walking, twice per stage. METHODS: Comparative study: within a 2-week period, 81 male soccer players at the Dutch premier league professional, premier league and 3rd division amateur levels performed the ISRT and the MMSRT. The total number of runs was scored for both tests. RESULTS: Only the ISRT distinguished between levels of competition. In general, the ISRT correlated moderately with the MMSRT for players of different levels of competition (rP: 0.52-0.83). CONCLUSIONS: In contrast to the MMSRT, the ISRT shows discriminative power for soccer players at different levels of competition supporting the validity of the ISRT for measuring endurance in a more soccer-specific way. As a result of the intermittent character of the ISRT the energetics of the ISRT and the MMSRT are not closely related. PMID- 15756162 TI - Effect of sleep deprivation on shuttle run score in middle-aged amateur athletes. Influence of initial score. AB - AIM: The aim of the present study was to investigate the effect of sleep deprivation on shuttle run score (corresponding to an estimated VO2 peak) in middle-aged amateur athletes. METHODS: Twenty-two athletes, training 9.2 hours a week (+/-4), performed a Leger and Gadoury shuttle test daily in a 4-day series that included a pre-test in day 0, 28 hours without sleep before test 1 (and 10 another hours after the test and before bedtime for a sleep deprivation of 38 hours), and then 10 hours of sleep before each of days 2 and 3. RESULTS: Shuttle run score showed no significant difference over the 4 days; however, the change in shuttle run score on day 2 was significantly correlated with the initial score. CONCLUSIONS: This result suggests that effect of sleep deprivation on shuttle run score is dependent on initial score and may encourage high level aerobic athletes to avoid sleep deprivation 2 days before a competition. PMID- 15756163 TI - Changes in the multi-joint kinematics and co-ordination after repetitive windsurfing pumping task. AB - AIM: This study investigated the change in kinematics after repetitive continuous maximal windsurfing pumping on a simulator by utilizing 3-D motion analysis. METHODS: Six competitive windsurfers exercised on a tailor-made simulator. The wind force was simulated by attaching the boom to the power head of a rowing ergometer. The exercise consisted of 3 sets of 3-minute maximum pumping. The motion of wrist, elbow, shoulder (on sagittal and frontal planes), waist, hip, knee and ankle joints were recorded by 2 video cameras. Kinematic analysis included the assessment of the change of the range of motion, movement speed (pulling phase and the whole pumping cycle), as well as the movement pattern of all tested joints. RESULTS: Modified motor strategy was documented by the significant drop of vertical excursion of the body centre of mass. Moreover, the ankle plantar flexors and dorsiflexors, mid-deltoid as well as wrist extensor and flexor, had significant change of movement according to the assessed parameters. Therefore, they may be the muscles that were more influenced than the other assessed muscles after continuous maximal pumping. The change of muscle co ordination may be a compensation mechanism to maintain work output. CONCLUSIONS: Findings in this study may suggest the importance of implementing specific muscle training program for the competitive windsurfers to increase the pumping capacity. PMID- 15756164 TI - The relationship between isometric contraction durations during hold-relax stretching and improvement of hamstring flexibility. AB - AIM: Previous research suggests proprioceptive neuromuscular facilitation (PNF) stretching techniques produce greater increases in range of motion than static or ballistic. The purpose of this study was to determine the isometric contraction hold time that best produces gains in hip range of motion after a 3-s (3-HR), 6 second (6-HR), and 10-s (10-HR) hold-relax PNF stretch. METHODS: The experimental design was a within subjects repeated measures 2 factor (baseline and treatment) by 3 factor (3-HR, 6-HR, 10-HR) ANOVA. The dependent variable was hamstring flexibility as measured in degrees of passive hip flexion with the knee extended. Measurements were taken in a university athletic therapy clinic. Sixty active individuals without history of knee or hip injury volunteered for this study. Each subject was randomly assigned to a 3-HR, 6-HR, or 10-HR group for the hold relax PNF stretch. All subjects were passively taken to the end range of motion 3 times and a goniometric measurement was taken. Each subject then performed the hold-relax PNF stretch 3 times with either a 3-HR, 6-HR, or 10-HR isometric contraction hold time and another goniometric measurement was taken. An average of each of these 3 trials was then calculated. RESULTS: The data analysis revealed a significant main effect difference (F(1,5 7)+/-633.97, p<.001, eta2=0.918) between the baseline and treatment measurements. There were no significant differences for the between groups (3-HR, 6-HR, 10-HR) main effect or for the interaction term. CONCLUSIONS: All 3 hold-time conditions produced significant gains in range of motion compared to baseline measurements. The application of the findings suggest that clinicians could choose any of the hold times and produce the same result to patient hip joint flexibility. PMID- 15756165 TI - Measuring energy expenditure in cardiac patients using the Body Media Armband versus indirect calorimetry. A validation study. AB - AIM: The purpose of this study was to compare differences in energy expenditure using the newly developed SenseWear Armband body monitor to indirect calorimetry in cardiac rehabilitation patients. METHODS: Twenty-four (62+/-8.1 years) patients completed steady state exercise for 8 minutes on 4 modes of exercise (arm ergometry, treadmill walking, recumbent stepping and rowing ergometry). Total cumulative kilojoules were recorded simultaneously by indirect calorimetry and the SenseWear Armband. Statistical analyses assessed the degree of agreement between the 2 measurement methods and 3 different versions of the SenseWear software. RESULTS: The correlations between indirect calorimetry and version 2.2 of the SenseWear armband for arm and rowing ergometry, the treadmill and recumbent stepper were r=0.90, r=0.67, r=0 .80 and r=0.74, respectively. There were no between method differences during arm ergometry (p<1.000) and the recumbent stepper (p<0.102). Bland and Altman plots revealed the greatest spread of scores for the rower and the treadmill. Between method differences were minimal when using the preliminary cardiac software. The correlations for arm and rowing ergometry, the treadmill and recumbent stepper were r=0.90, r=0.84, r=0.78 and r=0.82, respectively. CONCLUSIONS: The accuracy of the BodyMedia SenseWear Armband appears to be exercise modality dependent in those with heart disease when using software developed for the general population. Therefore, the information should be interpreted cautiously, particularly during treadmill walking and rowing. Initial experiments with cardiac-focused software are promising, but preliminary. PMID- 15756166 TI - High-intensity intermittent activities at school: controversies and facts. AB - In comparison to continuous aerobic type activity, little is known about high intensity intermittent physical activity in children. Repeated short-term high intensity activities (> maximal aerobic speed and <10 s) are more characteristic of the spontaneous physical activity of children. Recent studies have shown during repetitive bouts of sprints separated by short recovery intervals, that prepubescent children compared with adults are more able to maintain their performance without substantial fatigue. Moreover, repetitive runs at high velocities (near and higher than the maximal aerobic speed) separated by short recovery periods may elicit a high oxygen consumption in children. Several studies using interval training programmes for 7 weeks, twice a week for 30 min in physical education lessons showed that children's aerobic performance (maximal O2 uptake, maximal aerobic speed) could be enhanced. Training based on these repeated short-term high-intensity exercises could also improve children's anaerobic performance (short-term muscle power, strength and speed). Current evidence suggests that recovery from high-intensity exercises is faster in children than in adults and that repeated runs at high velocities separated by short recovery intervals can improve both aerobic and anaerobic performance. Although continuous aerobic type activity is more scientifically established as a training mode, repeated short-term high-intensity exercises in physical education programmes should be considered to enhance aerobic, as well as, anaerobic fitness in children. PMID- 15756167 TI - The effect of chronic and acute exercise on thymocyte apoptosis and necrosis in ovariectomized mice given dietary genistein. AB - AIM: At menopause, many women consume phytoestrogens instead of beginning hormone replacement therapy. Many also start exercise programs for health benefits. Genistein, a soy isoflavone with estrogen-like properties, induces lymphocyte apoptosis in vitro. Aerobic exercise also induces apoptosis in lymphoid cells. The present study was designed to determine the effect of chronic (wheel running, WR) and acute (treadmill) exercise on in vivo apoptosis of thymocytes using an animal model of menopause with supplementation from dietary genistein. METHODS: Using a randomized design, 99 ovariectomized B6D2F1 mice were fed 250 (GEN) or 0 (C) ppm genistein and given concurrent exercise (voluntary wheel running-WR; or WR followed by a bout of high intensity treadmill running-WR+TREAD) or remained sedentary (SED). After 21 days, mice were sacrificed for measurements of body weights, tissue weights, thymocyte apoptosis and necrosis by annexin-V FITC and propidium iodide staining and flow cytometry, DNA fragmentation by ELISA, and plasma estrogen concentrations by RIA. RESULTS: WR+TREAD mice had lower percentages of viable and higher percentages of apoptotic and necrotic cells from thymus compared with SED or WR conditions (p<0.001). WR resulted in greater DNA fragmentation in thymus cell lysates than in samples from SED mice (p<0.005). There were no differences in thymocyte apoptosis or DNA fragmentation between GEN and C mice, either independently or interactively with exercise. GEN mice tended to have greater wheel running activity than C mice (0.05120 km/w in training; 3 weeks of pre-race taper) and low (<80 km/w in training; 2 weeks of pre-race taper) training status groups. METHODS: Full blood counts, differential lymphocyte counts (CD3, CD4, CD8, CD19, CD56), serum cortisol, C-reactive protein (CRP) and creatine kinase (CK) were measured in blood samples donated 21 hours before and 16 hours after a 90 km ultramarathon. RESULTS: Despite significantly faster mean race finishing time (8.03 h vs 10.53 h; p<0.001) and greater percentage incidence (55.6% vs 40%) of post-race upper respiratory tract infection (URTI) in the highly trained group, these faster runners did not show evidence of a slower post-race recovery in terms of total leukocyte, neutrophil, total or differential lymphocyte counts (p>0.05). CRP concentrations were, however, markedly higher in the slower, less trained group (65.3+/-10.7 vs 38.3+/-5.9; p<0.01). CONCLUSIONS: Despite greater systemic evidence of post-race muscle inflammation and an acute phase response in the slower runners in a downhill ultramarathon race, the haematological recovery of well trained runners who undergo a 3-week taper period prior to the ultramarathon is not different to that in less trained runners who spend almost 3 hours longer on the road. The higher prevalence of post-race URTI symptoms in the fast, well trained group does not appear to be related to lymphocyte recovery in peripheral blood. PMID- 15756173 TI - Long-term endurance training induced changes in glucocorticoid receptors concentrations in rat and in man. AB - AIM: The responsiveness of target cells to glucocorticoids is directly related to the number and the functional state of intracellular glucocorticoid receptors (GR), which mediated the role of glucocorticoids. Although the effects of acute training on GR have been well characterized, less is known about the effects of chronic training on GR. The purpose of the present study was to describe the effects of long-term endurance training on GR. METHODS: Male Sprague-Dawley rats were divided into 2 groups at random, 7 as controls and exercisers (swimming 40 50 min/day for 80 days), respectively. All rats were sacrificed 24 hours after their last training session for measurements of the GR in liver and brain cytosol and thymus cells by radioligand binding assay with 3H-dexamethasone. This study was also carried out on 8 trained adult, 8 aged as well as 6 retired athletes, whose GR in peripheral leukocytes were analyzed. RESULTS: GR in hepatic cytosol and intact thymus cells of rats were significantly lower in exercise group than those in the controls. But the apparent dissociation constant (kd) was not changed. The GR in brain cytosol of rats indicated no significant reduction after training. The GR in peripheral leukocytes of the adult and aged athletes were also significantly lower than that in the controls. However, there was no significant difference between the GR in the retired athletes and that in the controls. CONCLUSIONS: This study demonstrated that long-term endurance training could lead to a decrease in GR and that the changes in GR were reversible during training. The possible mechanisms and the physiological significance of these changes need to be determined. PMID- 15756174 TI - Adverse effects of energy restriction on changes in immunoglobulins and complements during weight reduction in judoists. AB - AIM: A preliminary study to investigate the combined effects of dietary restriction and weight reduction through exercise on markers of immune function in college judoists before and after a single competition. METHODS: Forty-nine judoists participated in the study. Thirty-eight athletes combined exercise and dietary restriction (WR group), and 11 athletes did not require dietary restriction (EX group). Changes in anthropometric parameters, energy intake, concentrations of serum immunoglobulins and complements, and white blood cell counts were assessed at 4 time points: 20 days (pre-values), 4 days and 1 day before the competition, and 7 days after the competition. RESULTS: Compared with pre-values, the WR group exhibited significant decreases in body weight (-2.8 kg at 1 day before) and fat free mass (-1.7 kg at 1 day before); there were no changes in these variables in the EX group. The WR group exhibited significant decreases in IgG, IgM and C3 at 7 days after the competition (all p<0.01). In the EX group, significant decreases in IgM and C3 (both p<0.05) were observed at 7 days after the competition, though to a lesser degree than in the WR group. CONCLUSIONS: Energy restriction seemed to exacerbate alterations in immune markers such as immunoglobulin and complement induced by vigorous exercise at 7 days after a competition. Although the changed values were still within normal limits, we hypothesize that the potential cumulative effect of these changes over many competitions in 1 year might well induce abnormal levels with a possibly harmful clinical effect on judoists. PMID- 15756177 TI - Development of a perforated biodegradable interference screw. AB - PURPOSE: The objective was to develop a perforated biodegradable interference screw to allow for enhanced osseous implant integration without impairing screw stability during insertion. TYPE OF STUDY: Mechanical testing, followed by animal study. METHODS: At first, manual perforation of 8 x 23-mm biodegradable poly-(L co-D,L-lactide) interference screws was performed, using 3 different perforation patterns (clockwise spiral, counter-clockwise spiral, and parallel perforation), followed by torsional tests. Next, parallel perforated screws (n = 6) and unperforated control screws (n = 6) were applied to the proximal tibia of 12 sheep. The sheep were put down after 24 weeks and the screw site was examined histologically. Subsequently, molding of a parallel perforated screw followed by torsional tests was undertaken. RESULTS: The parallel perforated screw presented a torsional strength insignificantly different from the unperforated control screw and well beyond the reported maximum manual insertion torques of biodegradable interference screws in young human bone. When compared with the regular unperforated interference screw, the molded perforated screw exhibited a torsional strength of 91%, indicating a secure surgical application. In contrast to the unperforated screw, histologic evaluations revealed clear bone ingrowth into the perforations including the core of the perforated interference screw. CONCLUSIONS: Perforated, "cage-like" interference screws may be promising for the acceleration of osseous implant integration into the bone with a very low risk of screw breakage during insertion. CLINICAL RELEVANCE: To ameliorate osseous implant integration and possibly enhance ossification of former implant site in anterior cruciate ligament surgery. PMID- 15756178 TI - Primary stability of hamstring graft fixation with biodegradable suspension versus interference screws. AB - PURPOSE: During the early postoperative period, the stability of the fixation of a hamstring graft to the bone tunnel is the primary factor in limiting rehabilitation. The aim of this study was to evaluate if the initial fixation strength of a new suspension screw is comparable to that of the biodegradable interference screw fixation technique in the hamstring reconstruction of the anterior cruciate ligament (ACL). TYPE OF STUDY: Experimental laboratory study. METHODS: We evaluated the initial fixation strength of a biodegradable poly-L lactide/tri-calcium phosphate (PLLA/TCP) screw that suspended the graft in the bone tunnel and compared it with the strength of an interference screw for fixation of hamstring grafts in ACL reconstruction using bovine knees. Single cycle and cyclic loading tests were performed using a materials testing machine. RESULTS: The suspension screw provided a significantly higher yield load and ultimate failure load than the interference screw. There was no significant difference in the stiffness of both techniques. The typical failure mode for the suspension screw was fracture of the screw and for the interference screw it was slippage of the graft past the screw. In cyclic testing, both methods of fixation ran out to 1,000 cycles up to 250 N with a mean displacement of 2.6 mm (range, 1.8 to 3.3 mm) for the suspension screw and 4.1 mm (range, 2.3 to 6.0 mm) for the interference screw. Only the grafts fixed with the suspension screw survived a protocol with 1,000 cycles up to 400 N. CONCLUSIONS: Our biomechanical data suggest that hamstring graft fixation using a biodegradable PLLA/TCP suspension screw provides an alternative to interference screw fixation. CLINICAL RELEVANCE: Hamstring graft fixation using a suspension screw provides a reasonable alternative to interference screw fixation. PMID- 15756179 TI - Hamstring anterior cruciate ligament reconstruction: is it necessary to sacrifice the gracilis? AB - PURPOSE: The purpose of this study was to determine if there was a difference in the clinical results of anterior cruciate ligament (ACL) reconstruction when using the semitendinosus tendon (ST) alone versus the semitendinosus and gracilis (STG) construct. TYPE OF STUDY: Prospective randomized study. METHODS: We prospectively followed up a group of 97 patients who underwent reconstruction with either an ST graft (50 patients) or STG graft (47 patients). The patients were evaluated according to standard knee scales (IKDC, Noyes, Lysholm, Tegner), self-evaluation score (SANE), clinical findings, computerized knee laxity analysis, and functional tests. Isokinetic flexion, extension, and internal rotation-external rotation testing were also performed. The results were subjected to statistical analysis. RESULTS: We did not find any significant difference between the 2 groups according to the standard knee scores; self evaluation score; clinical findings; computerized knee laxity analysis; flexion, extension, and external rotation strengths; or functional tests. However, we did note that the internal rotation torque deficit was significantly higher in the STG group (P = .039). Likewise, the external-to-internal rotation ratio was significantly greater in the STG group (P = .006). CONCLUSIONS: Although there is not much clinical difference when using the ST alone versus the STG construct, internal rotation weakness following harvest of 2 tendons may need to be evaluated further. We suggest that, whenever possible, only 1 tendon should be used when performing anterior cruciate ligament reconstruction with hamstring tendons. LEVEL OF EVIDENCE: Level I, Randomized Controlled Study. PMID- 15756180 TI - Injury to the infrapatellar branch of the saphenous nerve in anterior cruciate ligament reconstruction: comparison of horizontal versus vertical harvest site incisions. AB - PURPOSE: Damage to the infrapatellar branch of the saphenous nerve (IPBSN) has been described for knee arthrotomy and arthroscopy. The true incidence of damage to this structure during anterior cruciate ligament (ACL) reconstruction has not been reported. Furthermore, the traditional vertical incision for central patellar tendon harvesting runs perpendicular to the course of this nerve. Therefore, a horizontal incision to avoid this potential complication was developed. TYPE OF STUDY: Case series. METHODS: A retrospective comparative study of ACL reconstructions was performed in 76 patients with central third patellar tendon as graft with a minimum of 2 years follow-up. Forty-two patients had a horizontal incision and 34 patients had a vertical incision. A questionnaire was used to evaluate function, satisfaction, and evidence of IPBSN damage. RESULTS: There was no difference between the 2 groups with regard to scar length, pain at rest, and pain with activity. Patients with a horizontal incision were more satisfied with the cosmetic appearance of their scar (P < .004). Twenty patients (59%) with a vertical incision had evidence of damage to the IPBSN as opposed to 18 patients (43%) with a horizontal incision. CONCLUSIONS: Although technically slightly more difficult, a horizontal incision may be a useful option for ACL graft harvesting to provide a more satisfactory scar with less risk of IPBSN damage. Regardless of the incision used, damage to the IPBSN is a potential complication and patients should be counseled of this preoperatively. LEVEL OF EVIDENCE: Level IV. PMID- 15756181 TI - Biomechanical comparisons of three different tibial tunnel directions in posterior cruciate ligament reconstruction. AB - PURPOSE: To investigate the effects of 3 different tunnel directions on the outcomes of posterior cruciate ligament (PCL) reconstruction surgery based on the forces exerted on the replacement ligament from a biomechanical point of view. The 3 tunnel directions in the proximal tibia are medial, central, and lateral. TYPE OF STUDY: Biomechanical study. METHODS: The forces exerted on the replaced PCL were calculated using finite element analyses as well as measurements from 6 cadavers. The results of the 3 surgical approaches were then compared. In the finite element analyses, the replaced ligament was assumed to have nonlinear elastic as well as viscoelastic properties. To simulate the overload in exercise, the femur was forced to move in the anterior direction abruptly while the tibia was held. From numerical analyses, the resultant forces, von Mises stresses, and maximum shear stresses on the replacement PCLs were calculated and compared. In the cadaveric study, a pressure-sensitive thin film was inserted between the replacement PCL and the killer turn area of the tibia. The color changes in films were evaluated using digital image processing in each case. RESULTS: The medial approach showed remarkably higher stresses and forces on the interface between the replaced PCL and the killer turn in both the numerical and cadaveric study. In contrast, the lateral approach showed the lowest stresses. CONCLUSIONS: The numerical and cadaveric studies indicate that the lateral approach is highly promising compared with the other approaches. CLINICAL RELEVANCE: The lateral approach has been shown to minimize stress concentration around the killer turn during in vitro experiments and a computer simulation of PCL reconstruction for long-term stability. The lateral approach technique appears to provide a promising clinical outcome in patients undergoing PCL reconstruction. PMID- 15756182 TI - Intercondylar notch stenosis in degenerative arthritis of the knee. AB - PURPOSE: To present a classification of intercondylar notch stenosis (IS) adjacent to the anterior cruciate ligament (ACL) in degenerative knee arthritis, to raise awareness of this disorder, to describe the arthroscopic findings, and to promote an organized approach to its treatment with favorable results. TYPE OF STUDY: Case series. METHODS: Of 362 arthroscopies in patients with gonarthrosis, we identified 122 knees in 96 patients (34%) with central knee pain and subjective instability without ACL laxity to determine the notch changes adjacent to the ACL. We followed a cohort of 69 knees in 64 patients, 47 female (73%) and 17 male (27%), excluding 53 knees in 32 patients for other symptomatic lesions, noncompliance with protocol, or loss to follow-up. The average patient age was 66 years (range, 53 to 78 years). Stenosis was classified as: type I, anterior; type II, lateral; type III, mixed; and type IV, massive. Diagnosis was determined by manipulation during arthroscopy to visualize impingement and was followed by notchplasty. Average follow-up was 26 months (range, 12 to 36 months). RESULTS: Type III was most common, appearing in 48% of knees. Type I was found in 29%, type II in 20%, and type IV in 3% of knees. Preoperatively, central pain occurred in all patients, being moderate in 40 knees (58%) and severe in 26 knees (38%), with diminished strength and subjective instability in all cases; only 42 (61%) had knee extension loss. Flexion contracture resolved in 81% of cases; 90% had good to excellent pain relief and 74% excellent relief of subjective instability, without significant complications. CONCLUSIONS: Intercondylar notch stenosis in the arthritic knee may be a cause of ACL damage, symptomatic instability, and loss of extension. A structured approach to diagnosis and treatment was beneficial in restoring more normal function for our patients and may prevent disease progression. LEVEL OF EVIDENCE: Level IV. PMID- 15756183 TI - Arthroscopic debridement of acute periprosthetic septic arthritis of the knee. AB - PURPOSE: To evaluate the efficacy of a treatment protocol including arthroscopic irrigation and debridement in resolving acute periprosthetic septic arthritis of the knee. TYPE OF STUDY: Retrospective review. METHODS: During a 1-year period, patients presenting with acute septic arthritis of the knee after total knee arthroplasty were screened for suitability for treatment with an arthroscopic debridement protocol consisting of (1) arthroscopic debridement and synovectomy with at least 12 L of antibiotic irrigant, (2) suction drainage until minimum output, (3) repeat arthroscopy for fever persisting beyond 48 hours, and (4) intravenous antibiotics for 6 weeks. Only previously well-functioning arthroplasties presenting within 7 days of symptom onset in patients without immunocompromising factors and with radiographically stable prosthetic components were selected for this treatment protocol. Five knees in 4 patients met the inclusion criteria and were examined clinically and radiographically at minimum 36-month follow-up. RESULTS: The mean follow-up interval was 41 months (range, 36 to 43 months). No knee had been revised or had revision planned for infection, although 1 had undergone tibial insert exchange for polyethylene wear. Follow-up radiographs showed no prosthetic loosening, and Knee Society scores averaged 88 (range, 75-95) in the 4 retained knees, 3 of which were subjectively thought to have returned to their preinfection level of functioning. All knees were infected with a single organism not considered highly virulent. No patients required oral suppressive antibiotics for their knees. CONCLUSIONS: Early aggressive arthroscopic debridement as part of a treatment protocol of acute periprosthetic knee septic arthritis with well-fixed and functioning implants can be an effective treatment option in selected cases. LEVEL OF EVIDENCE: Level IV study (no, or historical control group). PMID- 15756184 TI - Short-term analgesic effects of intra-articular injections after knee arthroscopy. AB - PURPOSE: To study the effects of 3 different intra-articular analgesic combinations on postoperative pain, and whether the timing of the injection, either preoperative or postoperative, had an effect on postoperative analgesia. TYPE OF STUDY: Prospective, randomized, double-blind clinical trial. METHODS: Patients undergoing knee arthroscopy performed by a single surgeon under general anesthesia were randomized into 6 study groups. Each patient received an intra articular injection 20 minutes before incision and at the end of the procedure. One injection contained the study medications, and the other contained normal saline solution (placebo). The patients, surgeon, and data collection personnel were blinded to the contents of the injections. Outcome measures included visual analogue pain scores at 0, 60, and 120 minutes after the procedure, and total recovery room fentanyl consumption. RESULTS: Fifty patients successfully completed the study protocol. Patients receiving combinations of morphine, bupivacaine, and epinephrine or bupivacaine and epinephrine yielded lower pain scores and narcotics consumption than patients receiving epinephrine alone, which was statistically significant irrespective of the timing of injection (P < .0001). Patients receiving the study medication preoperatively had significantly lower pain scores at the first measurement (t = 0) than those receiving the study medication postoperatively (P = .0343). There was no statistically significant effect of timing of the treatment medication administration at either 60 or 120 minutes postoperatively. Comparison of fentanyl consumption between groups receiving the treatment medication preoperatively versus postoperatively showed no significant difference. CONCLUSIONS: The combination of morphine, bupivacaine, and epinephrine, as well as the combination of bupivacaine and epinephrine provide excellent postoperative pain control when used either preoperatively or postoperatively in knee arthroscopy. There was a trend that patients receiving preoperative analgesic injections experienced superior pain control than did those injected postoperatively. LEVEL OF EVIDENCE: Level I, Randomized Controlled Trial. PMID- 15756185 TI - Intra-articular ropivacaine 0.75% and bupivacaine 0.50% for analgesia after arthroscopic knee surgery: a randomized prospective study. AB - PURPOSE: Intra-articular administration of local anesthetic solution provides analgesia after arthroscopic knee surgery. Bupivacaine is considered the gold standard local anesthetic in this indication, but ropivacaine, which is less toxic than bupivacaine, can consequently be administered in higher doses, potentially increasing the duration of analgesia. We compared the analgesic effect of intra-articular injection of ropivacaine 225 mg and bupivacaine 150 mg in patients undergoing arthroscopic surgery. TYPE OF STUDY: Double-blind, randomized prospective clinical study. METHODS: The study included 45 patients scheduled for knee meniscus repair under arthroscopy, who were allocated randomly into 3 groups to receive, intra-articularly, 30 mL of isotonic saline, bupivacaine 0.5%, or ropivacaine 0.75% solutions at the end of surgery. Postoperatively, pain was measured using a visual analog scale (VAS) at rest and on mobilization. Propacetamol was given when patients complained of pain. RESULTS: VAS scores at rest and on mobilization were higher in the saline group compared with the ropivacaine group (P = .006 and P = .01, respectively). No difference in VAS scores was documented between bupivacaine and the saline groups. The median delay between the intra-articular administration and propacetamol administration was shorter in the saline group (15 minutes; range, 15 to 120 minutes) compared with the ropivacaine group (240 minutes; range 15 to 720 minutes) but no difference was documented between the bupivacaine group (30 minutes; range, 15 to 360 minutes) and the other 2 groups. CONCLUSIONS: This level I study documents that intra-articular ropivacaine 0.75% provides better analgesia than bupivacaine 0.5% after knee arthroscopic surgery. LEVEL OF EVIDENCE: Level I. PMID- 15756186 TI - Arthroscopic treatment of anterolateral soft tissue impingement of the ankle: evaluation of factors affecting outcome. AB - PURPOSE: To evaluate the results for patients treated arthroscopically for anterolateral soft tissue impingement syndrome, to determine the factors affecting the outcome, and to report on a synovial shelf. TYPE OF STUDY: Retrospective clinical review. METHODS: Forty-one patients underwent operative arthroscopy for anterolateral impingement of the ankle between 1990 and 2001; the mean follow-up was 83.7 months (range, 21 to 152 months). There were 25 men and 16 women with an average age of 33.2 years (range, 15 to 63 years). All patients reported a history of inversion injury to the ankle. The most frequent preoperative complaints were tenderness localized to the anterolateral aspect of the ankle, swelling, crepitation, and pain at weight-bearing. All patients had failed to respond to at least 3 months of conservative treatment. The results were assessed according to Meislin's criteria and the American Orthopaedics Foot and Ankle Society (AOFAS) scoring table. For statistical analysis, the Mann Whitney U test was used where appropriate and the significance was set at P < .005. RESULTS: According to Meislin's criteria, there were excellent results in 21 patients, good in 16, fair in 2, and poor in 2. The mean AOFAS score was 89.6 points (range, 60-100) at follow-up. Four different soft tissue pathologies causing impingement were described. It has been statistically shown that cartilage damage located at the anterolateral region of the ankle and not advanced to the subchondral bone, and repeated inversion injuries had negative effects on clinical results at long-term follow-up. CONCLUSIONS: The arthroscopic diagnosis and treatment of anterolateral soft tissue impingement is a safe and effective method. Any combination of associated intra-articular pathologies, such as a chondral lesion or a new inversion injury of the ankle, after the arthroscopic procedure resulted in a poor outcome. LEVEL OF EVIDENCE: Level IV, Therapeutic Study. PMID- 15756187 TI - In vitro analysis of rotator cuff repairs: a comparison of arthroscopically inserted tacks or anchors with open transosseous repairs. AB - PURPOSE: The purpose of this study was to employ a cyclic loading protocol to compare rotator cuff repair strengths of arthroscopically inserted cuff tacks and suture anchors with the traditional open transosseous suture repair. TYPE OF STUDY: In vitro cadaveric analysis. METHODS: Full-thickness 1 x 3-cm rotator cuff defects were created in 25 fresh-frozen cadaveric shoulders, and were randomized to 1 of 4 repair groups: (1) open repair with transosseous sutures, (2) arthroscopic repair with 2 singly loaded suture anchors, (3) arthroscopic repair with 2 doubly loaded suture anchors, and (4) arthroscopic repair with cuff tacks. All repairs were cyclically loaded from 10 to 180 N, and the numbers of cycles to 50% (5-mm gap) and 100% (10-mm gap) failure were recorded. RESULTS: The number of cycles to 100% failure was significantly higher for the arthroscopic doubly loaded suture anchor repairs when compared with the (1) open transosseous suture repair (P = .009), (2) arthroscopic cuff tack repair (P = .003), and (3) arthroscopic singly loaded suture anchor repair (P = .02). Additionally, the number of cycles to 50% failure was significantly higher for all anchors versus open or tack repair (P = .03 for both). CONCLUSIONS: Immediate postoperative fixation of rotator cuff repairs with doubly loaded suture anchors was more stable than that provided by the open transosseous suture repairs, arthroscopic singly loaded suture anchors, or cuff tacks. However, additional evaluation is needed to examine the effects on the sustained strength of the repair throughout the healing process. CLINICAL RELEVANCE: These in vitro results indicate that superior immediate postoperative fixation of rotator cuff repairs may be achieved with the doubly loaded suture anchors. However, additional evaluation is needed to examine the effects on the sustained strength of the repair throughout the healing process. PMID- 15756188 TI - Arthroscopic versus mini-open rotator cuff repair: a cohort comparison study. AB - PURPOSE: To critically compare arthroscopic and mini-open rotator cuff repair. TYPE OF STUDY: Retrospective case control study. METHODS: Nine patients who had an arthroscopic rotator cuff repair (ARCR) were matched for age, gender, dominance, side of injury, history of trauma, duration of symptoms, and type of rotator cuff injury, with 12 patients who had a mini-open rotator cuff repair (MOR). Comparison included a preoperative and postoperative physical examination as well as a completed Simple Shoulder Test (SST) questionnaire at the latest follow-up at a minimum of 27 months. RESULTS: Both groups had significant reductions in pain scores (P < .01) and there was no significant difference in preoperative or postoperative active flexion or external rotation between both groups (P > .20). Although the ARCR group showed a significant improvement in strength (P < .01) and the MOR group did not, no patient had less than 4/5 strength. The impingement sign remained positive in 1 MOR patient, but all patients had a negative Jobe's test result. Pain and Tasks assessment by SST questionnaire showed that neither group had night pain or discomfort when using their arms overhead. There were no significant differences in the overall SST scores between groups. CONCLUSIONS: Because all patients in each group were satisfied with the procedure and there were no objective differences in outcome, we conclude that there is no difference in outcome between ARCR and MOR. Thus, the choice of one approach over the other is best based on surgeon or patient preference. LEVEL OF EVIDENCE: Level III, Retrospective Case Control Study. PMID- 15756189 TI - Diagnosis and treatment of chronic painful shoulder: review of nonsurgical interventions. AB - Chronic painful shoulder is a common complaint, yet its initial pathogenesis may have multiple etiologies as well as multiple and perhaps distinct primary or secondary etiologies responsible for its final clinical presentation. While the clinical presentation of pain and limitation of motion is easily defined, there are no standardized criteria for the differential diagnosis of various chronic shoulder pain syndromes, much less a consensus on their treatment. At present, there are limited nonsurgical options and medications approved for the treatment of chronic painful shoulder, and, with limited exceptions, there are no definitive conclusions about the efficacy of these therapies based on controlled clinical studies reported in the literature. There is a clear clinical need for local nonsurgical treatments that are safe and effective for chronic painful conditions. In this article, an attempt is made to review the diagnoses associated with chronic shoulder pain and some of the limited data that exist so as to suggest a therapeutic algorithm. PMID- 15756190 TI - Loose intra-articular body following anterior cruciate ligament reconstruction. AB - We report a case of intra-articular fracture of a bioabsorbable fixation device from the femoral tunnel in an anterior cruciate ligament reconstruction using a bone-tendon-bone graft. Thirteen months after successful reconstruction surgery, the patient experienced episodes of locking and medial joint pain. There was no history of trauma and no symptoms of instability or swelling. On revision arthroscopy, a fractured tip of a bioabsorbable RIGIDfix cross pin (Mitek, Westwood, MA) was identified in the medial compartment of the knee. There was a broad area of chondral erosion affecting the medial femoral condyle and a small defect to the medial tibial plateau where the loose body had been lodged. The bone-tendon-bone graft was intact without disruption. After arthroscopy, the patient was symptom free for 3 weeks but then developed further symptoms of locking. Magnetic resonance imaging showed another loose body within the knee. A repeat arthroscopy was performed 6 weeks after the earlier procedure and another piece of the polylactic acid RIGIDfix cross pin was removed, this time from the lateral gutter. This case raises concern about the potential for breakage and resultant loose body formation that may occur after bioabsorbable cross-pin fixation and, particularly, the associated chondral damage that can occur if early intervention is not conducted. PMID- 15756191 TI - Computed tomography-guided and arthroscopically controlled en bloc retrograde resection of a juxta-articular osteoid osteoma of the tibial plateau. AB - Osteoid osteoma represents approximately 10% to 11% of all the benign bone tumors. The localization at the articular and juxta-articular level of the knee is an atypical picture that causes both diagnostic and therapeutic problems. We present the case of an osteoid osteoma of the lateral tibial plateau just beneath the joint level. The lesion was removed by a rear-entry computed tomography (CT) guided drill under arthroscopic control and the bony defect filled with bone graft harvested from the proximal tibial metaphysis. Postoperative CT scans at 3 months, and 1 and 2 years were performed. The result of the kneeling test to evaluate donor-site morbidity 1 year after the surgery was negative. There were no immediate or delayed complications. Currently (2 years postoperatively), the patient has no pain and has gone back to his normal active daily lifestyle and routines. The follow-up CT scan 2 years after surgery showed complete excision of the lesion and perfect positioning of the bone graft. This new approach/technique enabled us to avoid damaging the proximal structures, to examine the lesion, and to fill the bony defect. Moreover, as shown by CT scan at follow-up, iatrogenic lesions to the cartilage of the tibial plateau were not reported. PMID- 15756192 TI - Arthroscopic bullet removal from the acetabulum. AB - Hip arthroscopy has been shown to offer minimally invasive access to the hip joint compared with standard open arthrotomy. The use of arthroscopy for diagnosing and treating disorders about the hip continues to evolve. The authors describe an arthroscopically assisted technique for the removal of a bullet lodged in the acetabulum of a patient who sustained a gunshot wound that entered the abdomen and traversed the rectum before ending up in the weight-bearing dome of the acetabulum. A number of issues led to the decision to use both arthroscopy and this specific technique. Most importantly was our desire to limit the amount of surrounding articular cartilage and local bone damage on removal. Minimizing the soft tissue dissection needed to access the bullet and keeping down our operative time also played a role in deciding to use this technique. We considered the risks of potential bullet fragmentation and migration, as well as a possible abdominal compartment syndrome before proceeding. This surgical technique afforded a very satisfactory outcome for this patient and serves as a model for others when encountering a similar injury pattern in a trauma patient. It is a procedure that can be performed safely, quickly, and with minimal complications for surgeons who have experience with arthroscopy of the hip joint. PMID- 15756193 TI - A simple technique for reconstruction of the medial patellofemoral ligament using a quadriceps tendon graft. AB - Numerous surgical procedures have been described for the treatment of recurrent patellar dislocation. Studies have shown that the primary pathoanatomy associated with lateral patellar dislocation is injury to the medial patellofemoral ligament (MPFL), which is the major soft-tissue restraint to lateral patellar translation. Recently, various procedures have been described that address the medial ligamentous injury inherent to lateral patellar dislocation. We present a simple technique for MPFL reconstruction using a quadriceps tendon graft. PMID- 15756194 TI - Minimally invasive "crescentic" imbrication of the medial patellofemoral ligament for chronic patellar subluxation. AB - Chronic patellar subluxation is one of the many causes of anterior knee pain. Nonoperative management of this disorder is successful in many patients, but surgical intervention may be necessary for refractory cases. We present a surgical technique for chronic patellar subluxation that is minimally invasive and is focused specifically on the medial patellofemoral ligament. PMID- 15756195 TI - Partial-thickness articular surface supraspinatus tears: a new transtendon suture technique. AB - The standard technique for repairing partial-thickness tears of the supraspinatus tendon includes completion of the lesion to a full-thickness tear. Partial articular-side supraspinatus tendon avulsions (PASTA) form a subgroup deserving special consideration. We present a transtendon suture technique that is able to preserve the intact tendon fibers and to achieve firm attachment of the tendon to the humeral footprint using 1 double-loaded bone anchor. The surgical technique is described in detail, and pitfalls and complications are noted. The clinical results of the first 22 consecutive patients are reported, showing an increase in the UCLA score from 17.1 to 31.2 points and a patient satisfaction rate of 91%. PMID- 15756196 TI - Traumatic rupture of the ligamentum teres as a source of hip pain. PMID- 15756204 TI - Untimely loss: lasting legacy. PMID- 15756205 TI - Inhaled nitric oxide for preterm infants: a therapy in search of an indication? The search continues. PMID- 15756206 TI - Growth failure in cystic fibrosis: a true need for anabolic agents? PMID- 15756207 TI - Chronic cholestasis and dyslipidemia: what is the cardiovascular risk? PMID- 15756208 TI - Modest lifestyle intervention attenuates the inflammatory state in children. PMID- 15756209 TI - Post-traumatic stress and its effect on health outcomes in children. PMID- 15756210 TI - Is there clinical evidence supporting the use of botanical dietary supplements in children? PMID- 15756211 TI - The safety and efficacy of nitric oxide therapy in premature infants. AB - OBJECTIVES: To assess the safety-efficacy balance of low-dose inhaled nitric oxide (iNO) in hypoxemic premature infants because no sustained beneficial effect has been demonstrated clearly and there are concerns about side effects. STUDY DESIGN: Eight hundred and sixty infants <32 weeks were randomized at birth to receive 5 ppm iNO or placebo when they presented with hypoxemic respiratory failure (HRF) defined by a requirement for mechanical ventilation, fraction of inspired oxygen (FIO 2 ) >40%, and arterio-alveolar ratio in oxygen (aAO 2 ) <0.22. The primary end point was intact survival at 28 days of age. RESULTS: Sixty-one of 415 infants presented with HRF and were compared with 84 of 445 controls who presented with HRF. There was no difference in the primary end point (61.4% in infants [23% with HRF who were treated with iNO] vs 61.1% in controls [21.4% in controls with HRF]; P = .943). For the infants with HRF who were treated with iNO, there was no significant difference from controls for intraventricular hemorrhage (IVH) (6% vs 7%), necrotizing enterocolitis (8% vs 6 %), or patent ductus arteriosus (PDA) (34% vs 37%). Compared with nonhypoxemic infants, the risk of bronchopulmonary displasia (BPD) increased significantly in HRF controls (OR = 3.264 [CI 1.461-7.292]) but not in infants with HRF who were treated with iNO (OR = 1.626 [CI 0.633-4.178]). CONCLUSIONS: iNO appears to be safe in premature infants but did not lead to a significant improvement in intact survival on day 28. PMID- 15756212 TI - Growth hormone treatment enhances nutrition and growth in children with cystic fibrosis receiving enteral nutrition. AB - OBJECTIVES: Impaired longitudinal growth and poor weight gain are common and important problems in children with cystic fibrosis. This study evaluates the hypothesis that adjunctive growth hormone (GH) therapy augments the growth response to nutritional supplementation. STUDY DESIGN: We recruited 18 prepubertal children who received enteral nutritional supplementation for at least 2 years before enrollment. Nine were randomly assigned to receive no GH for 1 year, followed by 1 year of GH. Nine were randomly assigned to receive 1 year of GH followed by a second year of GH. Measurements included height, weight, pulmonary function, lean tissue mass, bone mineral content, hospitalizations, outpatient antibiotic use, and caloric intake. RESULTS: Growth hormone resulted in significant improvement in height, weight, bone mineral content, lean tissue mass, and number of hospitalizations. Pulmonary function was similar at baseline. Absolute forced vital capacity and forced expiratory volume in 1 minute significantly increased in GH treatment, but there was no significant change in percent predicted pulmonary function. Caloric intake was similar in both groups during both years. CONCLUSIONS: These results suggest that GH is a useful for enhancing growth in children with cystic fibrosis receiving enteral nutritional supplementation. PMID- 15756213 TI - Evaluation of risk for atherosclerosis in Alagille syndrome and progressive familial intrahepatic cholestasis: two congenital cholestatic diseases with different lipoprotein metabolisms. AB - OBJECTIVES: To evaluate the risk for atherosclerosis in Alagille syndrome (AGS) and progressive familial intrahepatic cholestasis (PFIC) on the basis of lipoprotein metabolism and by ultrasonography. STUDY DESIGN: Five patients with AGS and 5 with PFIC, ages 3 to 4 years, were enrolled. Intimal-medial thickness and wall stiffness of the common carotid artery were examined by ultrasonography. Serum levels of lipids and lipoproteins were determined. Further, the chemical composition of LDL and its ability to transform macrophages into foam cells were determined. RESULTS: Intimal-medial thickness and wall stiffness were increased in patients with PFIC but not in patients with AGS. Total cholesterol, LDL cholesterol, HDL cholesterol, and lipoprotein X were remarkably increased in patients with AGS, whereas in patients with PFIC, an increase in triglyceride and a decrease in HDL cholesterol were the prominent findings. However, despite the normal LDL cholesterol level, oxidized LDL level was strikingly high in patients with PFIC. LDLs from patients with PFIC had high TG contents and exhibited high abilities to transform macrophages into foam cells. CONCLUSIONS: These findings suggest that patients with PFIC are at high risk for cardiovascular disorders involving atherosclerosis. PMID- 15756215 TI - Reduced adiponectin and HDL cholesterol without elevated C-reactive protein: clues to the biology of premature atherosclerosis in Hutchinson-Gilford Progeria Syndrome. AB - OBJECTIVES: Children with Hutchinson-Gilford Progeria Syndrome (HGPS) die of severe premature atherosclerosis at an average age of 13 years. Although the LMNA gene defect responsible for this "premature aging syndrome" has been identified, biological mechanisms underlying the accelerated atherosclerosis are unknown. We determined whether children with HGPS demonstrate abnormalities in known biomarkers for cardiovascular disease (CVD) risk. STUDY DESIGN: We quantified serum lipids, lipoproteins, C-reactive protein (CRP), and adiponectin in children with HGPS and age-matched control children. RESULTS: HDL cholesterol (P < .0001) and adiponectin (P < .001) concentrations decreased significantly with increasing age in HGPS but not in control children. There was a positive correlation between these variables in HGPS ( P < .0001) but not control children. Mean total cholesterol, LDL and HDL cholesterol, triglyceride, and median CRP levels were similar between HGPS and control children (all P > .05). CONCLUSIONS: Declining HDL cholesterol and adiponectin with advancing age may contribute to accelerated atherosclerotic plaque formation in HGPS. Several factors frequently associated with CVD risk in normal aging (elevated CRP, total and LDL cholesterol) showed no difference and are unlikely to influence CVD risk in HGPS. HDL and adiponectin may represent significant mediators and potential therapeutic targets for atherosclerosis in HGPS. PMID- 15756217 TI - Lifestyle-only intervention attenuates the inflammatory state associated with obesity: a randomized controlled study in adolescents. AB - OBJECTIVES: The primary goals were to understand the relationship among the inflammatory factors, C-reactive protein (CRP), interleukin-6 (IL-6), and fibrinogen, and indices of obesity in normoglycemic, insulin-resistant adolescents and to investigate the impact of a lifestyle-only intervention on these nontraditional risk factors for cardiovascular disease (CVD). STUDY DESIGN: Randomized controlled lifestyle-only intervention study in adolescents. Of the 21 adolescents studied, 15 obese subjects (body mass index [BMI] = 37.6 +/- 3.3 kg/m 2 ) were randomized to either a lifestyle intervention program or usual care. The lean controls were studied only at baseline. Analysis of variance (ANOVA) for repeated measures was used to study intervention effect and t test, one-way ANOVA, and discriminant function analysis for baseline comparisons. RESULTS: The intervention group maintained weight, whereas the control group gained weight (P = .02). A redistribution of body composition and a decrease in insulin resistance were observed. Elevated circulating concentrations of CRP, fibrinogen, and IL-6 were significantly reduced (all P or=3 spontaneous BMs per week with no fecal soiling, was similar in the control and experimental groups at 12 weeks (28/41 [68%] vs 31/43 [72%], respectively; P = .7) and at 24 weeks (27/41 [65%] vs 27/42 [64%], respectively; P = 1.0]. Groups also did not differ in their mean number of spontaneous BMs per week or episodes of fecal soiling per week at 4, 8, and 12 weeks. Adverse events and overall tolerance did not differ between groups. CONCLUSION: LGG, as dosed in this study, was not an effective adjunct to lactulose in treating constipation in children. PMID- 15756222 TI - A common mutation in the surfactant protein C gene associated with lung disease. AB - OBJECTIVE: To determine the contribution of the surfactant protein C (SP-C) I73T mutation to lung disease. STUDY DESIGN: Genomic DNA was obtained from 116 children with interstitial lung disease (ILD) or chronic lung disease of unclear cause and from 166 control subjects and was screened for the I73T mutation using an allele-specific polymerase chain reaction assay. RESULTS: The I73T mutation was found on 7 of 232 SP-C alleles from 7 unrelated children with ILD but was not found on 332 control SP-C alleles ( P < .01, Fisher exact test). The I73T mutation segregated with lung disease in one kindred with familial ILD. The I73T mutation was found in an asymptomatic parent from two different families with affected children consistent with variable penetrance, but it was not found in either asymptomatic parent of two other unrelated affected children consistent with a de novo mutation. Analysis of single nucleotide polymorphisms indicated diverse genetic backgrounds of the I73T alleles. Immunohistochemical analysis of lung tissue from an infant with the I73T mutation demonstrated normal staining patterns for proSP-B, SP-B, and proSP-C. CONCLUSIONS: These findings support the hypothesis that the I73T mutation predisposes to or causes lung disease. PMID- 15756223 TI - Pulmonary capillaritis in children: a review of eight cases with comparison to other alveolar hemorrhage syndromes. AB - OBJECTIVE: To review clinical, laboratory, and outcome characteristics of children diagnosed with pulmonary capillaritis (PC), a small-vessel vasculitis, presenting as diffuse alveolar hemorrhage (DAH), and to compare these findings with those for children with other alveolar hemorrhage syndromes. STUDY DESIGN: A retrospective chart review of patients who underwent a lung biopsy because of a clinical suggestion of pulmonary hemorrhage. RESULTS: PC was identified in 8 of 23 patients. In these patients, cough, crackles, and hypoxia were common. Alveolar infiltrates on radiography and anemia were present in 7 of 8 cases. Serologic evidence of a systemic vasculitis was present in 50% of patients. High dose corticosteroids proved effective in controlling alveolar hemorrhage in all cases. There were no presenting signs or symptoms that could differentiate patients with PC from those with non-immune-mediated alveolar hemorrhage. In general, patients with PC had a lower hematocrit and higher erythrocyte sedimentation rate (ESR). CONCLUSION: Children presenting with lower respiratory tract symptoms, chest x-ray abnormalities, and anemia should undergo evaluation for PC, as early initiation of immunosuppression can be lifesaving and organ sparing. No clinical signs to differentiate immune and non-immune-mediated alveolar hemorrhage were evident in this study. PMID- 15756225 TI - Genetic analyses in two extended families with deletion 22q11 syndrome: importance of extracardiac manifestations. AB - OBJECTIVES: Cardiovascular malformations (CVMs) are reported to be common (approximately 75%) in patients with deletion 22q11.2 (del22q11) syndrome. To better understand why deletions go unrecognized, we characterized the phenotype in deleted individuals in two large kindreds with particular emphasis on the presence or absence of CVM. STUDY DESIGN: After the diagnosis of del22q11 in two unrelated probands with CVM, we sequentially evaluated family members with clinical evaluation and cytogenetic analysis. RESULTS: Del22q11 was identified in 13 individuals; all exhibited characteristic dysmorphic facial features, but a CVM was present in only 6 of 13 (46%) individuals. CONCLUSIONS: We speculate that in the absence of CVM, diagnosis of del22q11 is hampered by a failure to recognize extracardiac features of the del22q11 syndrome spectrum. The data highlight the need for primary care physicians and specialists to familiarize themselves with the extracardiac stigmata of del22q11 to ensure timely diagnosis in all family members. PMID- 15756227 TI - Central nervous system hyperexcitability associated with glutamate dehydrogenase gain of function mutations. AB - OBJECTIVE: To describe seizure phenotypes associated with the hyperinsulinism/hyperammonemia syndrome (HI/HA), which is caused by gain of function mutations in the enzyme glutamate dehydrogenase (GDH). STUDY DESIGN: A retrospective review of records of 14 patients with HI/HA. RESULTS: Nine patients had seizures as the first symptom of HI/HA, and six had seizures in the absence of hypoglycemia. No electroencephalogram (EEG) background abnormalities were identified. In four patients, EEG recordings during seizures in the setting of normal blood glucose contained generalized epileptiform discharges. EEGs of three of these patients showed 0.5- to 2-second generalized irregular spike-and-wave discharge at 3 to 6 Hz corresponding to eye blinks, eye rolling, or staring. The EEG of the fourth patient consisted of 20 seconds of generalized regular spike and-wave discharge at 3 Hz in the clinical context of staring and unresponsiveness. In two patients, seizure control worsened with carbamezapine or oxcarbezapine treatment. CONCLUSIONS: In patients with HI/HA, generalized seizures are common and can occur in the absence of hypoglycemia. The drugs carbamazepine and oxcarbazepine should be used with caution for treatment. Pathogenesis of epilepsy in these patients may be related to effects of GDH mutations in the brain, perhaps in combination with effects of recurrent hypoglycemia and chronic hyperammonemia. PMID- 15756228 TI - Radiographic vertebral morphology: a diagnostic tool in pediatric osteoporosis. AB - OBJECTIVE: To assess the value of spinal radiographs in determining the significance of reductions in bone mass or density in chronically ill children. STUDY DESIGN: A pediatric scoring method for assessment of osteoporotic vertebral changes, developed on the basis of radiographs of 70 healthy controls and established adult scoring methods, was subsequently used to assess 32 pediatric patients with suspected secondary osteoporosis. Radiographic findings were correlated with bone mineral density (BMD), clinical data, and biochemistry. RESULTS: Thirty-two patients (median age 14.1 years) were included. Assessment of spinal radiographs with the developed scoring method found previously undiagnosed spinal compression deformities in 11 patients (34%) of whom 9 were asymptomatic and 8 had lumbar spine (size-corrected) BMD measurements within +/-2.0 SD of the age- and sex-specific norms. Fracture history and cumulative glucocorticoid (GC) dose did not differ between those with and without compression deformities. CONCLUSIONS: Vertebral compression fractures can be documented in a significant number of chronically ill children and are poorly predicted by single BMD measurements and clinical history. Assessment of vertebral morphology is recommended as an additional tool in the diagnostic workup of pediatric osteoporosis. PMID- 15756229 TI - Incidence and prevalence of HIV encephalopathy in children with HIV infection receiving highly active anti-retroviral therapy (HAART). AB - OBJECTIVE: To describe neurologic outcomes in children infected with HIV in the era of highly active anti-retroviral therapy (HAART), including rates of progressive HIV encephalopathy (PHE) and clinical sequelae among PHE survivors. STUDY DESIGN: Neurobehavior and school placement was assessed prospectively in the year 2000 in 126 children infected with HIV. PHE, developmental delay, and attention deficit disorder (ADHD) were the main outcome variables analyzed. Predictors of PHE were assessed in controlled analysis among age-matched controls. RESULTS: The rate of active PHE in 2000 was 1.6% (n = 2), and the prevalence of arrested PHE was 10% (n = 13). Residual motor and cognitive sequelae and need for special education was found in the majority of survivors. PHE relapse occurred in 3 (23%) children with previously arrested PHE. Viral load (VL) was the only significant factor associated with PHE. HIV or PHE was not associated with ADHD. Isolated developmental delay was not associated with HIV. CONCLUSIONS: PHE is an infrequent and reversible complication of HIV infection that responds to HAART and that may relapse if control of the virus is lost. Children with arrested PHE show higher rates of residual neurologic, cognitive, and scholastic impairments compared with children who never had PHE. Children with arrested PHE are the group of children with HIV infection most at risk for PHE, in the form of a relapse. PMID- 15756230 TI - Severe thrombocytopenia in a neonate with congenital HIV infection. PMID- 15756232 TI - Risk of hepatocellular carcinoma in liver mitochondrial respiratory chain disorders. AB - Mitochondrial respiratory chain disorders (MRCD) are a large group of disorders that can affect any organ besides muscles or the central nervous system. We report two children who presented with neonatal cholestasis and progressive cirrhosis, who subsequently developed hepatocellular carcinoma (HCC). This suggests a particular risk of degeneration in these patients and the importance of a regular screening for secondary liver cancer. Suggestion of HCC should lead to early liver transplantation, which was successful without tumor recurrence in the two patients. PMID- 15756233 TI - Liver transplantation as definitive treatment for a factor V Leiden mutation. AB - Liver transplantation (LT) was achieved for factor V Leiden-induced thrombophilia in a neonate with hepatic veno-occlusive disease. Initial LT was performed with a liver segment removed from a child with primary oxalosis. Four months later, a second, definitive LT was performed. The child remains well without recurrent thrombosis. PMID- 15756234 TI - Fatal varicella associated with selective natural killer cell deficiency. AB - A 2-year-old girl with recurrent severe varicella infections had a fatal outcome. Studies of cellular and humoral immunity were normal. No natural killer (NK) cells were detected, and NK activity was markedly decreased. The interleukin (IL)15/IL15R signaling pathway was intact. This case emphasizes the role of NK cells in controlling herpes viral infection. PMID- 15756236 TI - Esophageal diverticulum: a complication of histoplasmosis in children. AB - We report three boys, ages 8 to 14 years, who experienced dysphagia or chest pain while eating. In each patient, contrast esophagrams or esophagogastroduodenoscopy (EGD) demonstrated mid-esophageal traction diverticula, and serologic findings were compatible with acute or recent histoplasmosis. Diverticula appear to result from esophageal traction induced by inflammatory changes in adjacent infected lymph nodes. Antifungal and anti-inflammatory therapies were individualized in each case; all patients recovered fully. PMID- 15756237 TI - Parry-Romberg syndrome with fatal brain stem involvement. AB - We report the case of a 4 year-old boy with Parry-Romberg syndrome who had intractable seizures, progressive cerebral hemisphere atrophy, and fatal brain stem involvement. PMID- 15756238 TI - Kaposi varicelliform eruption in an infant. PMID- 15756239 TI - Resuscitation of newborn infants with 100% oxygen or air: a systematic review and meta-analysis. PMID- 15756240 TI - A randomized trial of a single dose of oral dexamethasone for mild croup. PMID- 15756241 TI - Are bedside features of shock reproducible between different observers? PMID- 15756242 TI - Effectiveness of adenotonsillectomy in children with mild symptoms of throat infections or adenotonsillar hypertrophy: open, randomised controlled trial. PMID- 15756245 TI - Retinal haemorrhages as proof of abusive head injury. PMID- 15756246 TI - Defining significant hydronephrosis. PMID- 15756247 TI - Widening sedation availability. PMID- 15756248 TI - Classification of cardiovascular malformations associated with neuroblastoma. PMID- 15756249 TI - From the many to the few: changes in urological cancer surgery provision. PMID- 15756250 TI - State-of-the-art management of nasopharyngeal carcinoma: current and future directions. AB - Nasopharyngeal carcinoma (NPC) is a distinct type of head and neck cancer. Approximately 70% of patients with newly diagnosed NPC present with locally advanced disease. Phase III clinical trials support the addition of chemotherapy to radiotherapy for the initial treatment of these patients. Once metastatic disease develops, practices become varied. Further experience needs to be gained with both targeted therapies and immunotherapy to gauge whether they will improve treatment outcomes in NPC. PMID- 15756251 TI - Trends in UK cancer trials: results from the UK Coordinating Committee for Cancer Research National Register of Cancer Trials. AB - We aimed to study trends in the design and conduct of randomised controlled trials (RCTs) in cancer in the UK, using the UK Coordinating Committee for Cancer Research (UKCCCR) National Register of Cancer Trials (NRCT). We conducted a descriptive survey of 520 UK RCTs in cancer that were registered on the UKCCCR NRCT. All trials had been initiated between 1971 and 2000. Trials on the NRCT have been conducted in a wide variety of cancer types, but with a third in breast (22%) or lung cancer (11%). They have largely been funded by the UK public and charity sectors. Overall, there has been a sustained rise in the total numbers of patients entering UK cancer trials over time with a trend towards larger, multicentre trials, greater recruitment targets and a marked reduction in the average time taken to complete trials. Trends in the design and conduct of noncommercial cancer RCTs from 1971 to 2000 are encouraging. It will be interesting to see how they develop in light of the implementation of recent national initiatives regarding cancer clinical trials in the UK. PMID- 15756252 TI - A phase I clinical and pharmacokinetic study of capecitabine (Xeloda) and irinotecan combination therapy (XELIRI) in patients with metastatic gastrointestinal tumours. AB - Capecitabine is a highly active oral fluoropyrimidine that is an attractive alternative to 5-fluorouracil in colorectal cancer treatment. The current study, undertaken in 27 patients with gastrointestinal tumours, aimed to assess the toxicity and potential for significant pharmacokinetic interactions of a combination regimen incorporating capecitabine with 3-weekly irinotecan (XELIRI). Irinotecan (200 and 250 mg m(-2)) was administered as a 90-min infusion on day 1 in combination with escalating capecitabine doses (700-1250 mg m(-2) twice daily) administered on days 2-15 of a 3-week treatment cycle. Pharmacokinetics were characterised on days 1 and 2 of the first two cycles. A total of 103 treatment cycles were administered. The principal dose-limiting toxicities were diarrhoea and neutropenia. Capecitabine 1150 mg m(-2) twice daily with irinotecan 250 mg m( 2) was identified as the maximum-tolerated dose and capecitabine 1000 mg m(-2) with irinotecan 250 mg m(-2) was identified as the recommended dose for further study. Analyses confirmed that there were no significant pharmacokinetic interactions between the two agents. The combination was clinically active, with complete and partial responses achieved in heavily pretreated patients. This study indicates that XELIRI is a potentially feasible and clinically active regimen in patients with advanced gastrointestinal cancer. PMID- 15756253 TI - Australian experience of a modified schedule of FOLFOX with high activity and tolerability and improved convenience in untreated metastatic colorectal cancer patients. AB - This study determined the efficacy and safety of a modified FOLFOX regimen that improved patient convenience without compromising oxaliplatin dose intensity. A total of 62 patients with previously untreated metastatic colorectal cancer were enrolled to receive, entirely as outpatients, 2-weekly cycles of oxaliplatin 100 mg m(-2) i.v. over 2 h, together with leucovorin 400 mg m(-2) over 2 h, 5 fluorouracil (5-FU) 400 mg m(-2), bolus, followed by a 46-h infusion of 5-FU at 2.4 g m(-2). Treatment was given until progression or unmanageable toxicity. In all, 61 patients received > or =one oxaliplatin dose and a median of 11 treatment cycles (range 1-20 cycles); 22 (36%) reported grade 3/4 neutropenia and 13 patients (21%) experienced grade 3 neurotoxicity; 16 patients (26%) discontinued treatment due to disease progression or death, 15 (25%) due to neurotoxicity and six (10%) due to haematological toxicity. Of the 56 eligible patients, complete or partial responses were observed in 29 or 52% (95% confidence interval 38-65%). Median progression-free survival was 8.2 months (7.1-9.9) and median overall survival was 18.7 months (14.0-23.4). In our experience, a modified schedule of FOLFOX improves convenience without compromising efficacy or toxicity. PMID- 15756255 TI - Primary tumour-vessel tumour-nodal tumour classification for patients with invasive ductal carcinoma of the breast. AB - There are many studies that show biological differences between invasive ductal carcinoma (IDC) with and without nodal metastasis, but no prognostic classification taking into consideration any biological differences between them is currently available. We previously investigated the histological characteristics that play an important role in tumour progression of IDCs according to their nodal status, and a new prognostic histological classification, the primary tumour-vessel tumour-nodal tumour (PVN) classification, was devised based on the histological characteristics of IDCs with and without nodal metastasis. Multivariate analyses using the Cox proportional hazard regression models were used to compare the ability of the PVN classification to predict tumour recurrence and death in 393 IDC patients based on the following histological classifications: (1) the pTNM classification, (2) the Nottingham Prognostic Index, (3) the modified Nottingham Prognostic Index, and (4) the histologic grade. In IDCs without nodal metastasis, only the PVN classification significantly increased the hazard rates (HRs) of tumour recurrence and death (P<0.05), independent of the hormone receptor status. Similarly, in IDCs with nodal metastases, only the PVN classification significantly increased the HRs of tumour recurrence and death (P<0.05), independent of the hormone receptor status. We conclude that the PVN prognostic histological classification is the best classification available for IDC of the breast. PMID- 15756254 TI - Adjuvant treatment with interleukin-2- and interferon-alpha2a-based chemoimmunotherapy in renal cell carcinoma post tumour nephrectomy: results of a prospectively randomised trial of the German Cooperative Renal Carcinoma Chemoimmunotherapy Group (DGCIN). AB - We conducted a prospectively randomised clinical trial to investigate the role of adjuvant outpatient immunochemotherapy administered postoperatively in high-risk patients with renal cell carcinoma. In total, 203 renal carcinoma patients' status post radical tumour nephrectomy were stratified into three risk groups: patients with tumour extending into renal vein/vena cava or invading beyond Gerota's fascia (pT3b/c pN0 or pT4pN0), patients with locoregional lymph node infiltration (pN+), and patients after complete resection of tumour relapse or solitary metastasis (R0). Patients were randomised to undergo either (A) 8 weeks of outpatient subcutaneous interleukin-2 (sc-rIL-2), subcutaneous interferon alpha2a (sc-rIFN-alpha2a), and intravenous 5-fluorouracil (iv-5-FU) according to the standard Atzpodien regimen (Atzpodien et al, 2004) or (B) observation. Two-, 5-, and 8-year survival rates were 81, 58, and 58% in the treatment arm, and 91, 76, and 66% in the observation arm (log rank P=0.0278), with a median follow-up of 4.3 years. Two, 5-, and 8-year relapse-free survival rates were calculated at 54, 42, and 39% in the treatment arm, and at 62, 49, and 49% in the observation arm (log rank P=0.2398). Stage-adapted subanalyses revealed no survival advantages of treatment over observation, as well. Our results established that there was no relapse-free survival benefit and the overall survival was inferior with an adjuvant 8-week-outpatient sc-rIL-2/sc-rIFN-alpha2a/iv-5-FU-based immunochemotherapy compared to observation in high-risk renal cell carcinoma patients following radical tumour nephrectomy. PMID- 15756256 TI - Insulin-like growth factor-1 (IGF1) genotype predicts breast volume after pregnancy and hormonal contraception and is associated with circulating IGF-1 levels: implications for risk of early-onset breast cancer in young women from hereditary breast cancer families. AB - BRCA1/2 mutations predispose to early-onset breast cancer, especially after oral contraceptive (OC) use and pregnancy. However, the majority of breast cancers might be due to more prevalent low-penetrance genes, which may also modify the risk in BRCA mutation carriers. The absence of the IGF1 19-repeat allele has been associated with high insulin-like growth factor-1 (IGF-1) levels during OC use. High IGF-1 levels are linked to early-onset breast cancer and larger breast volumes in the general population. The goal of this study was to elucidate the relationships between IGF1 genotype, early-onset breast cancer, breast volume, circulating IGF-1 levels and OC use in a prospective cohort of 258 healthy women < or =40 years old from high-risk breast cancer families. All women completed a questionnaire including information on reproductive factors and OC use. We measured the height, weight, breast volumes and plasma IGF-1 levels. IGF-1 levels were similar among parous and nulliparous women not using OCs. In all, 13% had no IGF1 19-repeat allele. There was an interaction between IGF1 genotype and OC use on IGF-1 levels (P=0.026) in nulliparous women and another interaction between IGF1 genotype and parity on breast volume (P=0.01). Absence of the 19-repeat allele was associated with high IGF-1 levels in nulliparous OC users and with larger breast volumes in parous women and OC users. Incident breast cancers were also more common in women without the 19-repeat allele (log rank P=0.002). Our results suggest that lack of the IGF1 19-repeat allele modifies IGF-1 levels, breast volume and possibly early-onset breast cancer risk after hormone exposure in young high-risk women. PMID- 15756257 TI - Adenoviral vector-mediated expression of a gene encoding secreted, EpCAM-targeted carboxylesterase-2 sensitises colon cancer spheroids to CPT-11. AB - CPT-11 (irinotecan or 7-ethyl-10[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin) is an anticancer agent in use for the treatment of colon cancer. In order to be fully active, CPT-11 needs to be converted into SN-38 (7 ethyl-10-hydroxycamptothecin) by the enzyme carboxylesterase (CE). In humans, only a minority of CPT-11 is converted to SN-38. To increase the antitumour effect of CPT-11 by gene-directed enzyme prodrug therapy, we constructed a replication-deficient adenoviral vector Ad.C28-sCE2 containing a fusion gene encoding a secreted form of human liver CE2 targeted to the surface antigen epithelial cell adhesion molecule (EpCAM) that is highly expressed on most colon carcinoma cells. By targeting CE2 to EpCAM, the enzyme should accumulate specifically in tumours and leakage into the circulation should be minimised. Ad.C28-sCE2-transduced colon carcinoma cells expressed and secreted active CE that bound specifically to EpCAM-expressing cells. In sections of three dimensional colon carcinoma spheroids transduced with Ad.C28-sCE2, it was shown that C28-sCE2 was capable of binding untransduced cells. Most importantly, treatment of these spheroids with nontoxic concentrations of CPT-11 resulted in growth inhibition comparable to treatment with SN-38. Therefore, Ad.C28-sCE2 holds promise in gene therapy approaches for the treatment of colon carcinoma. PMID- 15756258 TI - CD1a expression by Barrett's metaplasia of gastric type may help to predict its evolution towards cancer. AB - As emerging in the recent literature, CD1a has been regarded as a molecule whose expression may reflect tumour evolution. The aim of the present work was to investigate the expression of CD1a in a series of Barrett's metaplasia (BM), gastric type (GTBM), with and without follow-up, in order to analyse whether its expression may help to diagnose this disease and to address the outcome. Indeed, GTBM may be confused sometimes with islets of ectopic gastric mucosa and its evolution towards dysplasia (Dy) or carcinoma (Ca) could not be foreseen. We showed a significant higher expression of CD1a in GTBM than in both Dy and Ca; nevertheless, the number of positive GTBM was significantly lower in the group of cases that at follow-up underwent Dy or Ca. Our data address that CD1a may be a novel biomarker for BM and that its expression may help to predict the prognosis of this pathology. PMID- 15756259 TI - High viral loads of human papillomavirus predict risk of invasive cervical carcinoma. AB - High loads of human papillomavirus (HPV) 16 and HPV 18/45 increase the risk of developing invasive cervical carcinoma, revealing higher risk in percentiles of highest viral loads for HPV 16 (odds ratio (OR) 58.7, 95% confidence interval (CI) 21.9-151.4) compared to HPV 18/45 (OR 3.3, 95% CI 1.5-7.2). Thus, HPV load is a type-dependent risk marker for invasive carcinoma. PMID- 15756260 TI - Identification and analysis of tumour-associated antigens in hepatocellular carcinoma. AB - To identify tumour and tumour-associated antigens in patients with hepatocellular carcinoma (HCC) one may find potential diagnostic markers and immunotherapeutic targets. In the current study, 30 distinct antigens reactive with serum IgG from HCC patients were identified by serological analysis of cDNA expression libraries (SEREX). The mRNA expression patterns of 14 of these 30 antigens were altered in cancer as further revealed by cDNA microarray, with upregulation for nine and downregulation for five antigens. One of the upregulated antigens was cancer testis (CT) antigen (CAGE), which had been previously reported to be expressed exclusively in normal gametogenic tissues and aberrantly expressed in a variety of cancer cells. In our study, CAGE mRNA was expressed in 39.4% of HCC patients, 73.3% of patients with gastric cancer and 30.8% of patients with colorectal cancer. Antibodies against CAGE protein were detected in approximately 5.1% of the sera from HCC patients, 8.3% of that from gastric cancer patients and 7.3% of that from colorectal cancer patients. The relative high incidence of CAGE in cancer cells makes it a potential target for vaccine design. Another antigen of great interest is transgelin 2. The overexpression of transgelin 2 mRNA in a large per cent (69%) of HCC points to its potential as a diagnostic marker for HCC. PMID- 15756261 TI - Chromosome alterations in human hepatocellular carcinomas correlate with aetiology and histological grade--results of an explorative CGH meta-analysis. AB - All available comparative genomic hybridisation (CGH) analyses (n=31, until 12/2003) of human hepatocellular carcinomas (HCCs; n=785) and premalignant dysplastic nodules (DNs; n=30) were compiled and correlated with clinical and histological parameters. The most prominent amplifications of genomic material were present in 1q (57.1%), 8q (46.6%), 6p (22.3%), and 17q (22.2%), while losses were most prevalent in 8p (38%), 16q (35.9%), 4q (34.3%), 17p (32.1%), and 13q (26.2%). Deletions of 4q, 16q, 13q, and 8p positively correlated with hepatitis B virus aetiology, while losses of 8p were more frequently found in hepatitis C virus-negative cases. In poorly differentiated HCCs, 13q and 4q were significantly under-represented. Moreover, gains of 1q were positively correlated with the occurrence of all other high-frequency alterations in HCCs. In DNs, amplifications were most frequently present in 1q and 8q, while deletions occurred in 8p, 17p, 5p, 13q, 14q, and 16q. In conclusion, aetiology and dedifferentiation correlate with specific genomic alterations in human HCCs. Gains of 1q appear to be rather early events that may predispose to further chromosomal abnormalities. Thus, explorative CGH meta-analysis generates novel and testable hypotheses regarding the cause and functional significance of genomic alterations in human HCCs. PMID- 15756263 TI - Childhood leukaemia incidence and the population mixing hypothesis in US SEER data. PMID- 15756262 TI - Aberrant methylation of SPARC in human lung cancers. AB - SPARC (secreted protein acidic and rich in cysteine) is an extracellular Ca2+ binding matricellular glycoprotein associated with the regulation of cell adhesion and growth. We investigated loss of expression of SPARC gene and promoter methylation in lung cancers and correlated the data with clinicopathological features. We observed loss of SPARC expression in 12 of 20 (60%) lung cancer cell lines. Treatment of expression-negative cell lines with a demethylating agent restored expression in all cases. Methylation frequencies of SPARC gene were 55% in 20 lung cancer cell lines. Primary tumours had methylation at a rate of 69% (119 of 173), while nonmalignant lung tissues (n=60) had very low rates (3%). In lung adenocarcinomas, SPARC methylation correlated with a negative prognosis (P=0.0021; relative risk 4.65, 95% confidence interval 1.75 12.35, multivariate Cox's proportional-hazard model). Immunostaining revealed protein expression in bronchial epithelium (weak intensity) and in juxtatumoral stromal tissues (strong intensity) accompanied by frequent loss in cancer cells that correlated with the presence of methylation (P<0.001). Our findings are of biological interest and potentially of clinical importance in human lung cancers. PMID- 15756265 TI - Axillary recurrences after sentinel node (SLN) biopsy without complete axillary dissection in breast cancer patients. PMID- 15756268 TI - A prospective study of serum insulin-like growth factor-I (IGF-I), IGF-II, IGF binding protein-3 and breast cancer risk. AB - The associations between serum concentrations of insulin-like growth factor-I (IGF-I), IGF-II and IGF-binding proteins (IGFBP)-3 and risk of breast cancer were investigated in a nested case-control study involving 117 cases (70 premenopausal and 47 postmenopausal at blood collection) and 350 matched controls within a cohort of women from the island of Guernsey, UK. Women using exogenous hormones at the time of blood collection were excluded. Premenopausal women in the top vs bottom third of serum IGF-I concentration had a nonsignificantly increased risk for breast cancer after adjustment for IGFBP-3 (odds ratio (OR) 1.71; 95% confidence interval (CI): 0.74-3.95; test for linear trend, P=0.21). Serum IGFBP 3 was associated with a reduction in risk in premenopausal women after adjustment for IGF-I (top third vs the bottom third: OR 0.49; 95% CI: 0.21-1.12, P for trend=0.07). Neither IGF-I nor IGFBP-3 was associated with risk in postmenopausal women and serum IGF-II concentration was not associated with risk in pre- or postmenopausal women. These data are compatible with the hypothesis that premenopausal women with a relatively high circulating concentration of IGF-I and low IGFBP-3 are at an increased risk of developing breast cancer. PMID- 15756269 TI - A nested case-control study of the association of Helicobacter pylori infection with gastric adenocarcinoma in Korea. AB - In a nested case-control study of 86 cases of gastric adenocarcinoma in relation to Helicobacter pylori infection in the Korean Multi-center Cancer Cohort, the H. pylori IgG seropositivity was 83.7% and that of the 344 matched controls was 80.8%, with a matched odds ratio for H. pylori infection of 1.06 (95% CI, 0.80 1.40). PMID- 15756270 TI - Familial aggregation of lung cancer in a high incidence area in China. AB - To investigate whether lung cancer clusters in families in a high incidence county of China, an analysis was conducted using data on domestic fuel history and tobacco use for family members of 740 deceased lung cancer probands and 740 controls (probands' spouses). Lung cancer prevalence was compared among first degree relatives of probands and of controls, taking into account various factors using logistic regression and generalised estimating equations. First-degree relatives of probands, compared with those of controls, showed an excess risk of lung cancer (odds ratio (OR)=2.05, 95% confidence interval (CI): 1.68-2.53). Overall, female relatives of probands had a greater risk than did their male counterparts, and the risk was 2.90-fold for parents of probands as compared with parents of spouses. Female relatives of probands had 2.67-fold greater risk than female controls. Lung cancer risk was particularly marked among mothers (OR=3.78, 95% CI: 2.03-7.12). Having two or more affected relatives was associated with a 2.69-5.40-fold risk increase. The risk elevation was also found for other cancers overall. Results confirm previous findings of a genetic predisposition to lung cancer, and also imply that lung cancer may share a genetic background with other cancers. PMID- 15756271 TI - Optimisation of irinotecan dose in the treatment of patients with metastatic colorectal cancer after 5-FU failure: results from a multinational, randomised phase II study. AB - Although irinotecan 350 mg m(-2) is a standard option for relapsed/refractory advanced colorectal cancer, there is some evidence that suggests that a higher dose may be more effective, with acceptable tolerability, following 5 fluorouracil (5-FU). This study assessed the optimal dosing strategy for irinotecan, along with treatment efficacy and safety. A total of 164 patients with metastatic colorectal cancer progressing after failure on 5-FU or raltitrexed received either 350 mg m(-2) irinotecan (Group A; n=36) or 250, 350 or 500 mg m(-2), according to individual patient tolerance (Group B; n=62) or based on risk factor optimisation (Group C; n=66). There were no complete responses. There was a trend towards a higher overall response rate in Group B (13%) than in Groups A (8%) and C (9%). Tumour control growth rate was high in all three groups: 58% in group A, 60% in Group B and 50% in Group C. A total of 34% of patients in Group B and 9% in Group C were able to receive a dose of 500 mg m(-2). Median duration of response and time to progression were significantly longer in Groups A and B compared with Group C. No significant between-group differences for any adverse events were seen, although there was a small trend towards better tolerability in Group B. Individual dose escalation based on patient tolerance may allow more patients to receive a higher irinotecan dose without causing additional toxicity and can be an appropriate patient management strategy. PMID- 15756272 TI - K-ras mutation in the endometrium of tamoxifen-treated breast cancer patients, with a comparison of tamoxifen and toremifene. AB - The putative presence of a mutation in codon 12 of the K-ras gene was investigated in the endometrium of tamoxifen (TAM) and toremifene (TOR)-treated breast cancer patients. DNA was extracted from fresh cytologic samples of the endometrium in 86 TAM and 21 TOR-treated breast cancer patients. Mutations were detected by enriched PCR and an enzyme-linked mini-sequence assay (ELMA). K-ras mutation was found in 35 TAM-treated endometrial samples, and in only one TOR treated endometrium (P<0.003). In 24 premenopausal patients, K-ras mutation was found in seven (43.8%) of 16 patients with less than 47 months of TAM treatment, while none was found in eight patients with more than 48 months of TAM treatment (P<0.03). In 62 postmenopausal-amenorrheic patients, K-ras mutation was found in three (15.8%) of 19 patients with less than 23 months of TAM treatment, while it was found in 16 (61.5%) of 26 patients with 24-47 months of TAM treatment and nine (52.9%) of 17 patients with more than 48 months of TAM treatment (P=0.002). The presence of K-ras mutation is significantly influenced by the duration of TAM treatment and menstrual status of the patients. TOR may have a lower potential genotoxicity than TAM. PMID- 15756273 TI - Mutation analysis of three genes encoding novel LKB1-interacting proteins, BRG1, STRADalpha, and MO25alpha, in Peutz-Jeghers syndrome. AB - Mutations in LKB1 lead to Peutz-Jeghers syndrome (PJS). However, only a subset of PJS patients harbours LKB1 mutations. We performed a mutation analysis of three genes encoding novel LKB1-interacting proteins, BRG1, STRADalpha, and MO25alpha, in 28 LKB1-negative PJS patients. No disease-causing mutations were detected in the studied genes in PJS patients from different European populations. PMID- 15756274 TI - Dexamethasone-induced cisplatin and gemcitabine resistance in lung carcinoma samples treated ex vivo. AB - Chemotherapy for lung cancer not only has severe side effects but frequently also exhibits limited, if any clinical effectiveness. Dexamethasone (DEX) and similar glucocorticoids (GCs) such as prednisone are often used in the clinical setting, for example, as cotreatment to prevent nausea and other symptoms. Clinical trials evaluating the impact of GCs on tumour control and patient survival of lung carcinoma have never been performed. Therefore, we isolated cancer cells from resected lung tumour specimens and treated them with cisplatin in the presence or absence of DEX. Cell number of viable and dead cells was evaluated by trypan blue exclusion and viability was measured by the MTT-assay. We found that DEX induced resistance toward cisplatin in all of 10 examined tumour samples. Similar results were found using gemcitabine as cytotoxic drug. Survival of drug-treated lung carcinoma cells in the presence of DEX was longlasting as examined 2 and 3 weeks after cisplatin treatment of a lung carcinoma cell line. These data corroborate recent in vitro and in vivo xenograft findings and rise additional concerns about the widespread combined use of DEX with antineoplastic drugs in the clinical management of patients with lung cancer. PMID- 15756275 TI - The R72P P53 mutation is associated with familial breast cancer in Jewish women. AB - BRCA1/BRCA2 mutations account for a substantial proportion of familial breast cancer, but clearly mutations in additional genes exist, one candidate being the p53 gene. To evaluate its putative involvement in inherited predisposition to breast/ovarian cancer in Jewish high-risk women, mutational analysis of the p53 gene (exons 4-9) was carried out using exon-specific polymerase chain reaction followed by denaturing gradient gel electrophoresis (DGGE) analysis, complemented by DNA sequencing of abnormally migrating fragments. Overall, 132 Jewish breast cancer patient non-BRCA1/2 mutation carriers and 167 average risk controls (Ashkenazi (n=60), non-Ashkenazi (n=107)) were genotyped, and no inactivating p53 germline mutations were detected. Consistent migration abnormalities were noted in 167 fragments, 134 of which were shown to be the Arg72Pro polymorphism, whereas migration abnormalities in fragments containing exons 4 (n=2) and 6 (n=23) and introns 3 (n=4) and 9 (n=4) corresponded to five previously described polymorphisms. Allele distribution of the R72P missense mutation between ethnically diverse Jewish breast cancer cases and average risk controls showed significant differences: among non-Ashkenazi breast cancer cases, 62.5%, 33.3% and 4.2% were homozygous, heterozygous and homozygous for the Arg72, Arg72Pro and the Pro72 polymorphism, respectively, whereas for controls, the distribution was 22.4%, 65.4% and 12.2%, respectively (P=0.00052), and among Ashkenazi breast cancer cases, allele distribution was 68.5%, 29.6% and 1.9%, whereas for controls, the distribution was 50%, 40% and 10%, respectively (P=0.0125). We conclude that arginine homozygosity at codon 72 of the p53 gene is associated with a significant increased breast cancer risk in Jewish high-risk population. PMID- 15756276 TI - Phase I study of temozolomide plus paclitaxel in patients with advanced malignant melanoma and associated in vitro investigations. AB - The purpose of this study was to determine activity of temozolomide combined with paclitaxel or epothilone B in vitro, and to investigate the combination of temozolomide with paclitaxel in a Phase I clinical trial. Melanoma cell lines A375P and DX3 were treated with temozolomide and either paclitaxel or epothilone B. Combination indices were determined to assess the degree of synergism. In a clinical study, 21 patients with malignant melanoma were treated with increasing doses of temozolomide (orally, days 1-5), in combination with a fixed dose of paclitaxel (i.v. infusion day 1), followed by dose escalation of the latter drug. Cycles of treatment were repeated every 3 weeks. Pharmacokinetics of both agents were determined on day 1, with temozolomide pharmacokinetics also assessed on day 5. All three compounds were active against the melanoma cell lines, with epothilone B being the most potent. There was a strong degree of synergism between temozolomide and either paclitaxel or epothilone B. In the clinical study, no pharmacokinetic interaction was observed between temozolomide and paclitaxel. Dose escalation of both drugs to clinically active doses was possible, with no dose-limiting toxicities observed at 200 mg m(-2) day(-1) temozolomide and 225 mg m(-2) day(-1) paclitaxel. There were two partial responses out of 15 evaluable patients. One patient remains alive and symptom free at 4 years after treatment. Temozolomide and paclitaxel may be administered safely at clinically effective doses. Further evaluation of these combinations in melanoma is warranted. PMID- 15756277 TI - Epidermal growth factor receptor double targeting by a tyrosine kinase inhibitor (Iressa) and a monoclonal antibody (Cetuximab). Impact on cell growth and molecular factors. AB - Among the recent advances in the molecular targeted therapy of cancer, the applications focused on epidermal growth factor receptor (EGFR) are currently the most promising and the most advanced at clinical level. In view of the different modes of action of monoclonal antibodies and tyrosine kinase inhibitors (TKI), it is tempting to examine the effect of a combination between these two EGFR targeting approaches. It was the purpose of the present study to test this combination at experimental level by using two epidermoid human cell lines CAL 33 and CAL 39. As C225 (Cetuximab) and ZD1839 (Iressa) are, respectively, the most clinically advanced drugs in the category of anti-EGFR drugs, the experiments were performed using these two representative compounds. The combination of C225 and ZD1839 was antagonistic whatever the cell line considered. These antagonistic effects were corroborated by molecular changes in apoptosis (PARP) and EGFR signalling (phospho-p42-44). Drugs alone led to a diminution in EGFR levels, while their combination increased the cellular expression in EGFR. These data suggest that new and tempting treatment strategies on the EGFR target consisting in a double hit with a monoclonal antibody and a TKI must be considered with caution. PMID- 15756278 TI - Phase I/II study of oral etoposide plus GM-CSF as second-line chemotherapy in platinum-pretreated patients with advanced ovarian cancer. AB - The aim of this phase I/II study was to determine the maximum tolerated dose (MTD) and the dose-limiting toxicities of chronic oral etoposide given on days 1 10 followed by rescue with subcutaneous (s.c.) granulocyte-macrophage colony stimulating factor (GM-CSF) on days 12-19 as second-line chemotherapy in platinum pretreated patients (pts) with advanced ovarian carcinoma. Cohorts of three to six pts were treated with doses of oral etoposide from 750 mg m(-2) cycle(-1) escalated to 1250 mg m(-2) cycle(-1) over 10 days, every 3 weeks. Subcutaneous GM CSF, 400 mug once daily, days 12-19, was added if dose-limiting granulocytopenia was encountered. In total, 18 pts with a median Karnofsky index of 80% (range, 70 100%) and a median time elapsed since the last platinum dose of 10 months (range, 1-24 months), 30% of whom showed visceral metastases, were treated at four dose levels (DLs) of oral etoposide on days 1-10 of each cycle as follows: DL 1, 750 mg m(-2) cycle(-1), without GM-CSF, three pts; DL 2, 1000 mg m(-2) cycle(-1), without GM-CSF, three pts; DL 3, 1000 mg m(-2) cycle(-1), with GM-CSF, six pts; and DL 4, 1250 mg m(-2) cycle(-1), with GM-CSF, six pts. All pts were assessable for toxicity and 16 pts for response. Dose-limiting toxicity (DLT) was reached at DL 4 by three of six pts, showing World Health Organization (WHO) toxicity grade 4. One patient died from gram-negative sepsis associated with granulocytopenia grade 4. Two more pts developed uncomplicated granulocytopenia grade 4. Thus, we recommend that DL 3 can be used for further phase II evaluation (i.e. oral etoposide 1000 mg m(-2) cycle(-1), days 1-10, followed by s.c. GM-CSF 400 mug, days 12-19). The clinical complete or partial responses in each patient cohort were: DL 1, one of three pts; DL 2, one of three pts; DL 3, three of five pts; and DL 4, two of five pts. In conclusion, in this phase I/II study, we defined the MTD and the dose recommended for the therapy with oral etoposide given over 10 days followed by s.c. GM-CSF in platinum-pretreated patients with advanced ovarian cancer. Our data demonstrate encouraging activity of this regimen and strongly support its further investigation in a phase II study. PMID- 15756279 TI - Suicides among Danish cancer patients 1971-1999. AB - Compared to the general population, the suicide risk among Danish cancer patients diagnosed in 1971-1986 was increased by 50% for men and 30% for women. We updated the earlier study to evaluate both long-term and recent trends in the suicide risk. Cancer patients with a first cancer diagnosed between 1971 and 1999 in Denmark were followed-up for completed suicide through 1999. Excluding nonmelanoma skin cancer, 564 508 cancer patients were included and 1241 suicides observed. Both the standardised mortality ratio (SMR) of suicide relative to the general population and the suicide rates were analysed with Poisson regression methods. The overall SMR was increased to 1.7 (95% CI. 1.6-1.9) for men and 1.4 (95% CI: 1.3-1.5) for women. Following the cancer diagnosis, the suicide risk was highest in the first 3 months for men and between months 3 and 12 for women. The risk was higher for nonlocalized cancer and for cancers with perceived poor prognosis. Breast cancer patients had a higher risk than other cancer patients with similar good prognosis. The suicide rates among cancer patients decreased with calendar time, but less so than the rates in the general population. The suicide risk among cancer patients has not decreased as much as in the Danish population and reasons for this should be explored. Breast cancer might be believed by patients to be more life threatening than it is. Assessment and treatment of depression could improve the quality of life for cancer patients who suffer from unrecognised depressions and in turn reduce the risk of suicide in cancer patients. PMID- 15756280 TI - Aberrant DNA methylation associated with silencing BNIP3 gene expression in haematopoietic tumours. AB - Hypoxia is a key factor contributing to the progression of human neoplasias and to the development of resistance to chemotherapy. BNIP3 is a proapoptotic member of the Bcl-2 protein family involved in hypoxia-induced cell death. We evaluated the expression and methylation status of BNIP3 gene to better understand the role of epigenetic alteration of its expression in haematopoietic tumours. Methylation of the region around the BNIP3 transcription start site was detected in four acute lymphocytic leukaemia, one multiple myeloma and one Burkitt lymphoma cell lines, and was closely associated with silencing the gene. That expression of BNIP3 was restored by treatment with 5-aza2'-deoxycytidine (5-aza-dC), a methyltransferase inhibitor, which confirmed the gene to be epigenetically inactivated by methylation. Notably, re-expression of BNIP3 using 5-aza2-dC also restored hypoxia-mediated cell death in methylated cell lines. Acetylation of histone H3 in the 5' region of the gene, which was assessed using chromatin immunoprecipitation assays, correlated directly with gene expression and inversely with DNA methylation. Among primary tumours, methylation of BNIP3 was detected in five of 34 (15%) acute lymphocytic leukaemias, six of 35 (17%) acute myelogenous leukaemias and three of 14 (21%) multiple myelomas. These results suggest that aberrant DNA methylation of the 5' CpG island and histone deacetylation play key roles in silencing BNIP3 expression in haematopoietic tumours. PMID- 15756281 TI - Survival following lobectomy vs limited resection for stage I lung cancer: a meta analysis. AB - Extent of resection needed to treat lung cancer has long been an issue. The sole randomised controlled trial, reported by the Lung Cancer Study Group, advised against limited resection as standard surgery even for small peripheral non-small cell lung cancers (< or =3 cm), because of frequent local recurrences. Elsewhere, conflicting results have been reported from different institutions. We therefore conducted a meta-analysis of reported studies to compare survival of stage I patients between limited resection and standard lobectomy. A MEDLINE web search for computer-archived bibliographic data yielded 14 articles suitable for analysis. Combined survival differences (survival rate with lobectomy minus that with limited resection) at 1, 3, and 5 years after resection according to the DerSimonian-Laird random effects model were 0.7% (95% CI, -0.8 to 2.1; P=0.3659), 1.9% (95% CI, -3.7 to 7.4; P=0.5088), and 3.6% (95% CI, -0.4 to 10.5; P=0.3603), respectively. None of these survival differences were significant, indicating that survival after limited resection for stage I lung cancer was comparable to that after lobectomy. However, since interstudy heterogeneity was detected, caution is required in interpretation of the results. PMID- 15756282 TI - Reduced-intensity stem-cell transplantation for adult acute lymphoblastic leukemia: a retrospective study of 33 patients. AB - Efficacy of reduced-intensity stem-cell transplantation (RIST) for acute lymphoblastic leukemia (ALL) was investigated in 33 patients (median age, 55 years). RIST sources comprised 20 HLA-identical related donors, five HLA mismatched related, and eight unrelated donors. Six patients had undergone previous transplantation. Disease status at RIST was first remission (n=13), second remission (n=6), and induction failure or relapse (n=14). All patients tolerated preparatory regimens and achieved neutrophil engraftment (median, day 12.5). Acute and chronic graft-versus-host disease (GVHD) developed in 45 and 64%, respectively. Six patients received donor lymphocyte infusion (DLI), for prophylaxis (n=1) or treatment of recurrent ALL (n=5). Nine patients died of transplant-related mortality, with six deaths due to GVHD. The median follow-up of surviving patients was 11.6 months (range, 3.5-37.3 months). The 1-year relapse-free and overall survival rates were 29.8 and 39.6%, respectively. Of the 14 patients transplanted in relapse, five remained relapse free for longer than 6 months. Cumulative rates of progression and progression-free mortality at 3 years were 50.9 and 30.4%, respectively. These findings suggest the presence of a graft versus-leukemia effect for ALL. RIST for ALL is worth considering for further evaluation. PMID- 15756283 TI - Caspofungin as second-line therapy for fever of unknown origin or invasive fungal infection following allogeneic stem cell transplantation. AB - Caspofungin (CAS) is the first of a new class of antifungal agents, the echinocandins, that interfere with fungal cell wall synthesis by inhibition of glucan synthesis. Here, we report the results of 31 patients treated with CAS following allogeneic SCT. CAS was administered as a second-line agent to patients with invasive fungal infection (IFI) (n=15) or fever of unknown origin (n=16) who were recalcitrant to or intolerant of prior antifungal therapy. Unsuccessful first-line regimes included amphotericin B (n=17), liposomal amphotericin B (n=5), fluconazole (n=3), itraconazole (n=1), and voriconazole (n=2). All patients received concomitant immunosuppressive therapy for graft-versus-host disease. In 23 patients, cyclosporin A (CSA) and CAS were administered concurrently without any major side effects detected. Observed increases in GPT were not clinically significant. Normalization of serum creatinine and significant reductions in C-reactive protein were observed in response to CAS. Favorable outcome to CAS were documented in eight of 15 patients with IFI and in 15 of 16 patients with fever of unknown origin. CAS is a promising alternative in patients with IFI and fever of unknown origin in the setting of allogeneic SCT. PMID- 15756284 TI - Numerous nonclonal chromosomal aberrations arising in residual recipient hematopoietic cells following allogeneic bone marrow transplantation. AB - A young female patient in a second remission of acute lymphoblastic leukemia underwent bone marrow transplantation after total body irradiation and high-dose cytarabine from her HLA-matched brother. Following successful engraftment, mixed chimerism was seen 75 days post transplant. The karyotype contained numerous abnormalities in residual recipient cells. Chromosomes 1, 7, 13, and X were significantly more affected than other chromosomes. The high-frequency breakpoints identified were 1p22.2, 5q31.2, and 13q14.2. Some karyotypes specific for leukemia, such as t(9;22)(q34.1;q11.2) and t(8;21)(q22.2;q22.2), not seen with the original disease, were also present. As the frequency of aberrant chromosomes increased markedly with time, donor leukocytes were infused 14 months after BMT, which effectively eradicated the abnormal karyotypes. PMID- 15756286 TI - Targeting once-daily intravenous busulfan in combination with fludarabine before allogeneic hematopoietic cell transplantation. PMID- 15756285 TI - Outcome and long-term follow-up of alloreactive donor lymphocyte infusions given for relapse after myeloablative allogeneic hematopoietic stem cell transplantations (HSCT). AB - In order to study efficacy, toxicity and the long-term results of donor lymphocyte infusions (DLI), we retrospectively analyzed DLI given for relapse after conventional allogeneic hematopoietic stem cell transplantation (HSCT) in 30 patients with a median delay of 107.5 months after transplant and 58 months after DLI. After DLI, 15 patients established full donor chimerism, three patients developed grade III and one grade IV acute GVHD. A total of 15 patients achieved a disease response. Among the 14 patients with chronic myeloid leukemia (CML), 11 are alive at the last follow-up: five are in complete molecular response (CMR) and two in complete cytogenetic response (CCR) with no other intervention after DLI, three in CMR after imatinib mesylate given after DLI and one in complete hematological response after imatinib mesylate and reduced intensity conditioning allogeneic SCT performed after DLI. At the time of the last follow-up, 19 (63%) patients died and 11 (37%) remain alive. The 3-year probability of survival for the entire population, CML patients and non-CML patients, was 60, 93, 62% after transplantation, and 48, 80 and 48% after DLI, respectively. A multivariate analysis demonstrated a significantly worse survival rate after transplantation for female recipients, advanced disease and acute leukemia before transplantation. PMID- 15756287 TI - Red cell fragmentation preceding a renal crisis in a patient with systemic sclerosis undergoing haematopoietic stem cell transplantation. PMID- 15756288 TI - Successful bone marrow harvest during pregnancy. PMID- 15756291 TI - A DNA recombination-based approach to eliminate papillomavirus infection. AB - At present, no treatments exist that effectively target and eliminate papillomaviruses (PVs) from infected cells or prevent its replication. We are employing a strategy to prevent virus replication in PV-infected cells through the conditional expression of the herpes simplex virus type 1 thymidine kinase (TK) gene. Expression of TK in this system is expected to be triggered by a homologous recombination event between the endogenous PV genome and a nonexpressing TK gene cassette. Recombination between these two DNAs is expected to change the nonexpressing cassette into a form that expresses TK. Various constructs were generated to express the TK in the above manner. Transfection of cell lines with a TK nonexpressing plasmid did not result in TK production due to alternative splicing and polyadenylation site selection. However, cotransfection of cell lines with PV plasmids along with the above TK construct containing short segments of PV sequences resulted in a recombination event that led to TK expression as shown by Northern and Western blot analyses. We also developed a TK expression cassette utilizing an adeno-associated virus (AAV) vector. Delivery of the cassette by AAV to PV-infected cells resulted in TK expression, and ganciclovir treatment resulted in efficient killing of these cells. PMID- 15756294 TI - Iron absorption by human subjects from different iron fortification compounds added to Thai fish sauce. AB - OBJECTIVES: (a) To measure iron absorption by human subjects from citric acid stabilized fish sauce fortified with ferrous sulfate, ferric ammonium citrate or ferrous lactate and (b) to identify the effect of added citric acid (3 g/l) on iron absorption from ferrous sulfate fortified fish sauce. DESIGN: Iron absorption from the intrinsically labeled compounds was determined via erythrocyte incorporation of isotopic labels ((57)Fe and (58)Fe) using a randomized crossover design. In three separate absorption studies, 10 adult women each consumed a basic test meal of rice and vegetable soup seasoned with isotopically labeled, iron fortified fish sauce. RESULTS: Iron absorption was significantly lower from ferrous lactate and from ferric ammonium citrate fortified fish sauce than from ferrous sulfate fortified fish sauce. Fractional iron absorption (geometric mean; -1s.d., +1s.d.) was 8.7(3.6; 21.4)% for ferrous lactate compared to 13.0(5.4; 31.4)% from ferrous sulfate, P = 0.003 (study 1) and 6.0(2.5; 14.3)% from ferric ammonium citrate relative to 11.7(4.4; 30.7)% from ferrous sulfate, P < 0.001, in study 2. Citric acid added at a molar ratio of approximately 2.5 to iron had no effect on iron absorption from ferrous sulfate (study 3). Iron absorption in the presence of citric acid was 14.1(6.4; 30.8)% compared to 12.0(5.8; 24.7)% in its absence (P = 0.26). CONCLUSIONS: Iron absorption was 50-100% higher from ferrous sulphate fortified fish sauce than from fish sauce fortified with ferric ammonium citrate or ferrous lactate. In the presence of citric acid as a chelator, ferrous sulfate would appear to be a useful fortificant for fish sauce. SPONSORSHIP: International Atomic Energy Agency (IAEA), Vienna, Austria. PMID- 15756295 TI - Evolutionary genetics: the human brain -- adaptation at many levels. PMID- 15756296 TI - Twin study of genetic and aging effects on X chromosome inactivation. AB - To investigate the genetic influence on X chromosome inactivation and on age related skewing of X inactivation, in particular, we analysed the X inactivation pattern (XIP) in peripheral blood cells from 118 young monozygotic (MZ) twin pairs (18-53 years), 82 elderly MZ twin pairs (55-94 years), 146 young dizygotic (DZ) twin pairs (20-54 years) and 112 elderly DZ twin pairs (64-95 years). Elderly twins had a higher frequency of skewed X inactivation (34%) than young twins (15%) (P<0.001). Our data suggest that the increase in skewing occurs after age 50-60 years. The intraclass correlation was 0.61 and 0.58 in young and elderly MZ twin pairs, and 0.08 and 0.09 in young and elderly DZ twin pairs. Biometric analysis showed that dominant genetic effects accounted for 63 and 58% of the variance of XIP in the young and elderly twin pairs, respectively. The dominant genetic effect and the shared environment for monochorionic MZ twins may explain the high intraclass correlation for the MZ twin pairs compared to the DZ twin pairs. We did not observe a significant decrease in the intraclass correlation in elderly MZ twins compared to young MZ twins, which would be expected if age-related skewing were due to stochastic factors. We conclude that the increased skewing with age implies that a genetically dependent selection of blood cells take place. PMID- 15756297 TI - High frequencies of Y chromosome lineages characterized by E3b1, DYS19-11, DYS392 12 in Somali males. AB - We genotyped 45 biallelic markers and 11 STR systems on the Y chromosome in 201 male Somalis. In addition, 65 sub-Saharan Western Africans, 59 Turks and 64 Iraqis were typed for the biallelic Y chromosome markers. In Somalis, 14 Y chromosome haplogroups were identified including E3b1 (77.6%) and K2 (10.4%). The haplogroup E3b1 with the rare DYS19-11 allele (also called the E3b1 cluster gamma) was found in 75.1% of male Somalis, and 70.6% of Somali Y chromosomes were E3b1, DYS19-11, DYS392-12, DYS437-14, DYS438-11 and DYS393-13. The haplotype diversity of eight Y-STRs ('minimal haplotype') was 0.9575 compared to an average of 0.9974 and 0.9996 in European and Asian populations. In sub-Saharan Western Africans, only four haplogroups were identified. The West African clade E3a was found in 89.2% of the samples and the haplogroup E3b1 was not observed. In Turks, 12 haplogroups were found including J2*(xJ2f2) (27.1%), R1b3*(xR1b3d, R1b3f) (20.3%), E3b3 and R1a1*(xR1a1b) (both 11.9%). In Iraqis, 12 haplogroups were identified including J2*(xJ2f2) (29.7%) and J*(xJ2) (26.6%). The data suggest that the male Somali population is a branch of the East African population - closely related to the Oromos in Ethiopia and North Kenya - with predominant E3b1 cluster gamma lineages that were introduced into the Somali population 4000-5000 years ago, and that the Somali male population has approximately 15% Y chromosomes from Eurasia and approximately 5% from sub-Saharan Africa. PMID- 15756298 TI - Homocysteine levels--before and after methionine loading--in 51 Dutch families. AB - Elevated levels of homocysteine are a risk factor for vascular disease, thrombosis, neural tube defects and dementia. The 677C>T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene appears to be the most important single determinant of plasma homocysteine concentration. In the current study, we estimated heritability and fit a series of models of inheritance for both fasting and postmethionine-load homocysteine levels in the HOFAM-study (HOmocysteine in FAMilies study), which included 306 participants from 51 pedigrees, ascertained through a hyperhomocysteinemic proband. The crude heritability was 21.6% for fasting and 67.5% for postloading homocysteine. After adjustment for MTHFR 677C>T genotype, heritability dropped to 5.2 and 63.9%, respectively. Segregation analysis revealed that a nongenetic model with equal transmission was the best fitting and most parsimonious model for fasting homocysteine levels, while a two distribution, Mendelian model with residual familial correlation was best for postmethionine-load homocysteine levels. This study shows that postload homocysteine levels have a stronger genetic determination than do fasting homocysteine levels. The heritability of postload homocysteine levels were not strongly affected by adjustment for MTHFR 677C>T genotype, in contrast to fasting homocysteine levels. Further studies are needed to identify the genes responsible for the inheritance of postload homocysteine levels. PMID- 15756299 TI - Schistosomal hepatic fibrosis and the interferon gamma receptor: a linkage analysis using single-nucleotide polymorphic markers. AB - A minority of individuals infected with the parasite Schistosoma mansoni develops hepatic fibrosis. HLA studies in Egypt and a candidate gene search in a Sudanese population indicate that the host's genetics contribute to disease susceptibility. In an Egyptian community, 32.7% of individuals 11 years and older had significant fibrosis by WHO ultrasound criteria. Linkage to 10 candidate genes was tested using 89 affected sibling pairs from 40 pedigrees in this community. The candidates included genes that initiate fibrosis, participate in collagen synthesis, or control collagen degradation. Two to four single nucleotide polymorphisms (SNPs) were genotyped per locus, and 188 individuals were genotyped at 48 markers. Model-free modified Haseman-Elston analysis identified linkage to a SNP in the interferon gamma receptor locus (P=0.000001). There was also weak evidence for linkage to the interleukin 13-4 region and tissue growth factor beta 1. PMID- 15756300 TI - FISH of supernumerary marker chromosomes (SMCs) identifies six diagnostically relevant intervals on chromosome 22q and a novel type of bisatellited SMC(22). AB - Supernumerary marker chromosomes (SMCs) are frequently found at pre- and postnatal cytogenetic diagnosis and require identification. A disproportionally large subset of SMCs is derived from the human chromosome 22 and confers tri- or tetrasomy for the cat eye chromosomal region (CECR, the proximal 2 Mb of chromosome 22q) and/or other segments of 22q. Using fluorescence in situ hybridization (FISH) and 15 different DNA probes, we studied nine unrelated patients with an SMC(22) that contained the CECR. Five patients showed the small (type I) cat eye syndrome (CES) chromosome and each one had the larger (type II) CES chromosome, small ring chromosome 22, der(22)t(11;22) extrachromosome, and a novel type of bisatellited SMC(22) with breakpoints outside the low-copy repeats (LCRs22). By size and morphology, the novel bisatellited SMC(22) resembled the typical (types I and II) CES chromosomes, but it might have been associated with the chromosome 22q duplication syndrome, not CES. This SMC included a marker from band 22q12.3 and conferred only one extra copy each of the 22 centromere, CECR, and common 22q11 deletion area. There has been no previous report of a bisatellited SMC(22) predicting the chromosome 22q duplication syndrome. Accounting for the cytogenetic resemblance to CES chromosomes but different makeup and prognosis, we propose naming this an atypical (type III) CES chromosome. In this study, we found six distinct intervals on 22q to be relevant for FISH diagnostics. We propose to characterize SMCs(22) using DNA probes corresponding to these intervals. PMID- 15756301 TI - A y(+)LAT-1 mutant protein interferes with y(+)LAT-2 activity: implications for the molecular pathogenesis of lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is an inherited aminoaciduria caused by defective cationic amino acid (CAA) transport at the basolateral membrane of epithelial cells in the intestine and kidney. The SLC7A7 gene, mutated in LPI, encodes the y(+)LAT-1 protein, which is the light subunit of the heterodimeric CAA transporter in which 4F2hc is the heavy chain subunit. Co-expression of 4F2hc and y(+)LAT-1 induces the y(+)L activity. This activity is also exerted by another complex composed of 4F2hc and y(+)LAT-2, the latter encoded by the SLC7A6 gene and more ubiquitously expressed than SLC7A7. On the basis of both the pattern of expression and the transport activity, y(+)LAT-2 might compensate for CAA transport when y(+)LAT-1 is defective. By expression in Xenopus laevis oocytes and mammalian cells, we functionally analysed two SLC7A7 mutants, E36del and F152L, respectively, the former displaying a partial dominant-negative effect. The results of the present study provide further insight into the molecular pathogenesis of LPI: a putative multiheteromeric structure of both [4F2hc/y(+)LAT-1] and [4F2hc/y(+)LAT-2], and the interference between y(+)LAT-1 and y(+)LAT-2 proteins. This interference can explain why the compensatory mechanism, that is, an increased expression of SLC7A6 as seen in lymphoblasts from LPI patients, may not be sufficient to restore the y(+)L system activity. PMID- 15756303 TI - About the origin and development of hereditary conventional renal cell carcinoma in a four-generation t(3;8)(p14.1;q24.23) family. AB - Conventional renal cell carcinoma (CRCC) may appear in families with germline translocations involving chromosome 3, although a recurrent responsible gene has not been found. We recently described a family with CRCC and a constitutional t(3;8)(p14.1;q24.23), and we demonstrated that no genes were disrupted by the translocation breakpoints. In order to investigate the genetic origin and features of the CRCC tumors that occurred in this family, we have extended the pedigree up to four generations, and analyzed peripheral blood samples from 36 members, CRCC tumors, normal renal tissues, and a gastric tumor. (1) By means of comparative genomic hybridization (CGH), we have detected loss of the derivative chromosome carrying 3p in all CRCC but not in the corresponding normal renal tissue. In addition, by means of the fluorescence in situ hybridization technique, we have observed that not all tumoral cells lose the der(3p), which suggests that, previous to this loss, another hit should occur to initiate the transformation of normal into tumoral cells. (2) All known mechanisms of inactivation of the candidate von Hippel-Lindau (VHL) gene have been studied in the tumors, detecting alterations in 65% of them. This confirms that inactivation of the VHL gene is not always required to develop CRCC, and that (an)other suppressor gene(s) on 3p could be involved. (3) We discard FHIT as an alternative pathway to VHL. We have not found new candidate regions along 3p by using a 1-Mb resolution array-based CGH. (4) The tumorigenesis mechanism of a second gastric tumor developed in the probandus is different from that of CRCC. PMID- 15756302 TI - COMP mutation screening as an aid for the clinical diagnosis and counselling of patients with a suspected diagnosis of pseudoachondroplasia or multiple epiphyseal dysplasia. AB - The skeletal dysplasias are a clinically and genetically heterogeneous group of conditions affecting the development of the osseous skeleton and fall into the category of rare genetic diseases in which the diagnosis can be difficult for the nonexpert. Two such diseases are pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED), which result in varying degrees of short stature, joint pain and stiffness and often resulting in early onset osteoarthritis. PSACH and some forms of MED result from mutations in the cartilage oligomeric matrix protein (COMP) gene and to aid the clinical diagnosis and counselling of patients with a suspected diagnosis of PSACH or MED, we developed an efficient and accurate molecular diagnostic service for the COMP gene. In a 36-month period, 100 families were screened for a mutation in COMP and we identified disease causing mutations in 78% of PSACH families and 36% of MED families. Furthermore, in several of these families, the identification of a disease-causing mutation provided information that was immediately used to direct reproductive decision making. PMID- 15756305 TI - The influence of chronic exposure to antipsychotic medications on brain size before and after tissue fixation: a comparison of haloperidol and olanzapine in macaque monkeys. AB - It is unclear to what degree antipsychotic therapy confounds longitudinal imaging studies and post-mortem studies of subjects with schizophrenia. To investigate this problem, we developed a non-human primate model of chronic antipsychotic exposure. Three groups of six macaque monkeys each were exposed to oral haloperidol, olanzapine or sham for a 17-27 month period. The resulting plasma drug levels were comparable to those seen in subjects with schizophrenia treated with these medications. After the exposure, we observed an 8-11% reduction in mean fresh brain weights as well as left cerebrum fresh weights and volumes in both drug-treated groups compared to sham animals. The differences were observed across all major brain regions (frontal, parietal, temporal, occipital, and cerebellum), but appeared most robust in the frontal and parietal regions. Stereological analysis of the parietal region using Cavalieri's principle revealed similar volume reductions in both gray and white matter. In addition, we assessed the subsequent tissue shrinkage due to standard histological processing and found no evidence of differential shrinkage due to drug exposure. However, we observed a pronounced general shrinkage effect of approximately 20% and a highly significant variation in shrinkage across brain regions. In conclusion, chronic exposure of non-human primates to antipsychotics was associated with reduced brain volume. Antipsychotic medication may confound post-mortem studies and longitudinal imaging studies of subjects with schizophrenia that depend upon volumetric measures. PMID- 15756304 TI - Behavioral tolerance to lysergic acid diethylamide is associated with reduced serotonin-2A receptor signaling in rat cortex. AB - Tolerance is defined as a decrease in responsiveness to a drug after repeated administration. Tolerance to the behavioral effects of hallucinogens occurs in humans and animals. In this study, we used drug discrimination to establish a behavioral model of lysergic acid diethylamide (LSD) tolerance and examined whether tolerance to the stimulus properties of LSD is related to altered serotonin receptor signaling. Rats were trained to discriminate 60 microg/kg LSD from saline in a two-lever drug discrimination paradigm. Two groups of animals were assigned to either chronic saline treatment or chronic LSD treatment. For chronic treatment, rats from each group were injected once per day with either 130 microg/kg LSD or saline for 5 days. Rats were tested for their ability to discriminate either saline or 60 microg/kg LSD, 24 h after the last chronic injection. Rats receiving chronic LSD showed a 44% reduction in LSD lever selection, while rats receiving chronic vehicle showed no change in percent choice on the LSD lever. In another group of rats receiving the identical chronic LSD treatment, LSD-stimulated [35S]GTPgammaS binding, an index of G-protein coupling, was measured in the rat brain by autoradiography. After chronic LSD, a significant reduction in LSD-stimulated [35S]GTPgammaS binding was observed in the medial prefrontal cortex and anterior cingulate cortex. Furthermore, chronic LSD produced a significant reduction in 2,5-dimethoxy-4-iodoamphetamine stimulated [35S]GTPgammaS binding in medial prefrontal cortex and anterior cingulate cortex, which was blocked by MDL 100907, a selective 5-HT2A receptor antagonist, but not SB206553, a 5-HT2C receptor antagonist, indicating a reduction in 5-HT2A receptor signaling. 125I-LSD binding to 5-HT2A receptors was reduced in cortical regions, demonstrating a reduction in 5-HT2A receptor density. Taken together, these results indicate that adaptive changes in LSD stimulated serotonin receptor signaling may mediate tolerance to the discriminative stimulus effects of LSD. PMID- 15756306 TI - Antianxiety and antidepressive behavior produced by physiological estradiol regimen may be modulated by hypothalamic-pituitary-adrenal axis activity. AB - Variations in estradiol (E(2)) may influence expression of stress-related anxiety and depression symptoms among women. Effects of E(2) and stress on anxiety and depressive behavior were investigated using an animal model. E(2) was administered subcutaneously (0, 2, 5, 10, 20, 50 mug/rat) to ovariectomized rats 2 days before testing. In experiment 1, open field (anxiety), elevated plus maze (anxiety), or forced swim test (depressive) behavior was evaluated following 20 min of restraint or no such stressor. Rats administered 5 or 10 mug E(2), which produced physiological plasma E(2) concentrations, showed significantly less anxiety and depressive behavior and lower corticosterone levels compared to vehicle, lower, or higher E(2) dosages. Restraint stress prior to behavioral testing attenuated the antianxiety and antidepressive effects of 5 or 10 mug E(2). In experiment 2, effects of adrenalectomy or sham surgery and vehicle or corticosterone replacement in their drinking water on behavior and neuroendocrine measures of rats administered 0, 10, or 50 mug E(2) were examined. E(2), 10 mug, compared to vehicle or 50 mug, reduced anxiety and depressive behavior of sham and adrenalectomized rats administered the low dosage of corticosterone, but not vehicle or the high dosage of corticosterone, suggesting that there may be an optimal level of corticosterone necessary for E(2) to exert these effects. Together, these data suggest that E(2) may have dose-dependent effects on anxiety and depressive behavior of female rodents, which may depend on the tone of the hypothalamic-pituitary-adrenal axis. PMID- 15756310 TI - "Smart dust": nanostructured devices in a grain of sand. AB - The term "smart dust" originally referred to miniature wireless semiconductor devices made using fabrication techniques derived from the microelectronics industry. These devices incorporate sensing, computing and communications in a centimetre-sized package. This article discusses the construction of much smaller silicon-based systems, using the tools of nanotechnology. The synthesis of millimetre- to micron-sized functional photonic crystals made from porous silicon is described. It is shown how the various optical, chemical, and mechanical properties can be harnessed to perform sensing, signal processing, communication and motive functions. PMID- 15756311 TI - Combinatorial libraries - from solution to 2D microarrays. AB - Enzymatic modifications of split and mix libraries were followed by "pulling down" onto a 2-dimensional DNA microarray, via PNA tagging; this allowed complete library interrogation of all members of the split and mix library. PMID- 15756312 TI - Catalytic asymmetric synthesis of enantiopure isoprenoid building blocks: application in the synthesis of apple leafminer pheromones. AB - The first catalytic asymmetric procedure capable of preparing all 4 diastereoisomers (ee > 99%, de > 98%) of a versatile saturated isoprenoid building block was developed and the value of this new method was demonstrated in its application to the concise total synthesis of two pheromones. PMID- 15756313 TI - Air-stable Ir-(P-Phos) complex for highly enantioselective hydrogenation of quinolines and their immobilization in poly(ethylene glycol) dimethyl ether (DMPEG). AB - An air-stable catalyst system Ir-(P-Phos) catalyst was found to be highly effective in the asymmetric hydrogenation of quinoline derivatives. The catalyst immobilized in DMPEG was efficiently recovered and reused eight times, retaining reactivity and enantioselectivity. PMID- 15756314 TI - A new macrocycle demonstrates ditopic recognition properties. AB - The synthesis and binding properties of a new macrocycle is reported. The host, comprised of three basic pyridines, four hydrogen bond accepting carbonyls, and two hydrogen bond donating amide groups, binds mono-alkyl ammonium salts in a manner that is dependent on the counter-ion of the ammonium guest. PMID- 15756315 TI - A chiral layered Co(II) coordination polymer with helical chains from achiral materials. AB - A layered coordination polymer Co(PDC)(H2O)2.H2O containing two helical chains was synthesized, and the resultant crystals were not racemic as evidenced by the observation of strong signals in vibrational circular dichroism (VCD) spectra. PMID- 15756316 TI - Synthesis of poly(ethyleneimine)s-silica hybrid particles with complex shapes and hierarchical structures. AB - Shaped silica constructed by silica fiber with axial poly(ethyleneimine)s (PEIs) filaments was available simply from rapid hydrolytic condensation of tetramethoxysilane (TMOS) upon aggregates of PEIs with different architecture. PMID- 15756317 TI - Reduction of dipyrido-[3,2-a:2',3'-c]-phenazine (dppz) by photolysis in ethanol solution. AB - Photolysis of dipyrido-[3,2-a:2',3'-c]-phenazine (dppz) (1) in ethanol solution leads to the formation of 9,14-dihydrodipyridophenazine (2), which has been characterised by detailed NMR analysis, UV/VIS absorption spectroscopy, and theoretical calculations which reveal that its red colour is due to a low-lying intramolecular charge transfer state. PMID- 15756318 TI - Formation, stability, and reactivity of a mononuclear nonheme oxoiron(IV) complex in aqueous solution. AB - A mononuclear nonheme oxoiron(IV) complex bearing a pentadentate N5 ligand was prepared in aqueous solution; the pH dependence of its stability and reactivities was reported along with the mechanistic details of sulfide oxidation by the oxoiron(IV) species. PMID- 15756319 TI - High-efficiency red electrophosphorescence based on neutral bis(pyrrole)-diimine platinum(II) complex. AB - Efficient red electroluminescence from the excimer or oligomer of neutral phosphorescent bis(pyrrole)-diimine Pt(II) complex has been achieved with maximum external quantum efficiency, luminous efficiency, power efficiency and brightness of 6.5%, 9.0 cd A(-1), 4.0 lm W(-1) and 11 100 cd m(-2), respectively. PMID- 15756320 TI - Preparation and crystallographic characterization of C60Cl24. AB - C60Cl24 has been synthesized by the chlorination of C60 with VCl4 or C60Br24 with SbCl5; the X-ray single crystal structure of C60Cl24.2Br2 confirmed the molecular T(h) symmetry in good agreement with the IR data and theoretical calculations. PMID- 15756321 TI - A high spin molecular square based on square pyramidal Co(II) and tetrahedral Mn(II) centers: [{MnIICl2}2{CoII(triphos)(CN)2}2]. AB - A high spin molecular square of square pyramidal Co(II) and tetrahedral Mn(II) corners exhibiting an overall S= 4 ground state has been crystallized and its magnetic properties investigated. PMID- 15756322 TI - Unexpected conversion of a hexacyanometallate to a homoleptic nitrile complex with triphenylborane substituents. AB - The homoleptic, triphenylborane adduct of the hexacyanochromate anion was structurally characterized and found to be a product in which all six cyanide ligands have undergone isomerization to nitrile groups. PMID- 15756323 TI - The effect of spacer chain length on ion binding to bidentate alpha,omega diamines: contrasting ordering for H+ and Li+ ion affinities. AB - Electrospray ionisation mass spectrometry studies and quantum chemical calculations indicate that bidentate ligation of Li+ ion to the diamines leads to symmetric bridging and exhibits contrasting relative affinity orderings compared to that of proton for alpha,omega-diamines. PMID- 15756324 TI - Organosilicate-surfactant lamellar mesophase with molecular-scale periodicity in the silicate layers. AB - The synthesis of lamellar mesophases of organosilicate-surfactant composites with periodicity within the silicate layers due to periodic arrangement of phenylene- and biphenylylene-silica moieties is reported. PMID- 15756325 TI - Mild metal-free syn-stereoselective ring opening of activated epoxides and aziridines with aryl borates. AB - A conceptually new, simple and practical method for the syn-nucleophilic displacement of aryl and vinyl epoxides and aryl aziridines with (substituted) phenols, using aryl borates as activating nucleophiles under neutral conditions, is reported. PMID- 15756326 TI - Preferential CO oxidation promoted by the presence of H2 over K-Pt/Al2O3. AB - In preferential CO oxidation in H2-rich gas, K-Pt/Al2O3(K/Pt = 10) was very effective in decreasing CO concentration below 10 ppm in the 375-410 K range, and the turnover frequency of the K-Pt/Al2O3 was 20 times as high as that of Pt/Al2O3 at 353 K; furthermore, the activity of CO oxidation was promoted drastically by the presence of H2. PMID- 15756327 TI - Lanthanide 8-hydroxyquinoline-based podates with efficient emission in the NIR range. AB - The novel hydroxyquinoline-containing tetrapodal ligand forms water soluble and stable chelates and is a good sensitizer of the NIR luminescence of its Nd(III) and Yb(III) complexes; its easy synthesis opens the way for potential biomedical applications. PMID- 15756328 TI - Wet-chemical templateless assembly of metal nanowires from nanoparticles. AB - We describe a new, simple, room-temperature wet-chemical approach for assembling Au and Ag nanoparticles into nanowire networks, without the use of lithographic templates. Five to 35 nm-diameter nanowires passivated with a thin organic layer were synthesized by mechanically agitating a biphasic liquid mixture of an aqueous hydrosol containing the nanoparticles, and toluene. Nanowire structure and surface chemistry are discussed based on electron microscopy, UV-visible spectroscopy and thermogravimetric analyses. PMID- 15756329 TI - Coordination chemistry of the bis(trifluoromethylsulfonyl)imide anion: molecular interactions in room temperature ionic liquids. AB - Room temperature ionic liquids composed of bis(trifluoromethylsulfonyl)imide anions and 1,3-ethylmethylimidazolium (EMI) cations are shown to stabilize monomeric ligand deficient transition metal complexes via four distinct binding modes: monodentate nitrogen or oxygen coordination and/or bidentate oxygen oxygen' or nitrogen-oxygen coordination (eta1-N, eta1-O, eta2-O,O' and eta2-N,O). PMID- 15756330 TI - Lysine-enediyne conjugates as photochemically triggered DNA double-strand cleavage agents. AB - Statistical analysis of DNA-photocleavage by two types of lysine-enediyne conjugates confirms that more double-strand breaks are produced than can be accounted for by coincident single-strand breaks. PMID- 15756331 TI - A simple and convenient route to prepare poly(vinylidene fluoride trifluoroethylene) copolymer nanowires and nanotubes. AB - Poly(vinylidene fluoride trifluoroethylene) copolymer nanowires and nanotubes have been prepared for the first time via a high temperature (99% ees for aromatic aldehydes and 99% ees for alphatic aldehydes were offered by the present procedure. PMID- 15756333 TI - Nanoplatelet-based reconstructed hydrotalcites: towards more efficient solid base catalysts in aldol condensations. AB - Rehydration of Mg-Al hydrotalcite in the liquid phase using ultrasounds or a high stirring speed leads to nanoplatelets with surface areas of 400 m(2) g(-1), displaying catalytic activities in aldol condensations up to 8 times higher than the best catalytic system reported in the literature. PMID- 15756334 TI - Nickel catalysed asymmetric cycloisomerisation of diethyl diallylmalonate. AB - Cationic nickel catalysts with monodentate phosphoramidites and Wilke's azaphospholene as ligands are highly regio- and enantioselective catalysts for the cycloisomerisation of diethyl diallylmalonate. PMID- 15756335 TI - Alkynes as activators in the nickel-catalysed addition of organoboronates to aldehydes. AB - Alkynes act not as substrates but as co-catalysts in the presence of a nickel catalyst, an organoboronate and an aldehyde to promote the addition reaction between the substrates in combination with H2O. PMID- 15756336 TI - Enhancement in the reducibility of cobalt oxides on a mesoporous silica supported cobalt catalyst. AB - The silylation of SBA-15 enhances the reducibility of cobalt oxides on a SBA-15 supported cobalt catalyst, and consequently increases the catalytic activity for Fischer-Tropsch synthesis of hydrocarbons from syngas and selectivity for longer chain products. PMID- 15756337 TI - Hexyl-substituted oligothiophenes with a central tetrafluorophenylene unit: crystal engineering of planar structures for p-type organic semiconductors. AB - Rigidification has been achieved in thiophene-tetrafluorophenylene architectures through strong S...F and H...F intramolecular interactions; the resulting materials are promising candidates for p-type organic field effect transistors. PMID- 15756339 TI - Selection experiments with dynamic combinatorial libraries: the importance of the target concentration. AB - Experimental data are presented which demonstrate that the target concentration has a strong influence on the outcome of selection experiments with a dynamic mixture of self-assembled receptors. PMID- 15756338 TI - Bisindoles containing a 2,1,3-benzothiadiazole unit: novel non-doping red organic light-emitting diodes with excellent color purity. AB - A new class of bisindole-based red fluorescent materials was prepared and used as non-doping red emitters to give the devices emitting saturated red colour at about 650 nm with a chromaticity coordinate of (x = 0.64, y = 0.34), matching the CIE 1931 standard red color very well. PMID- 15756340 TI - A simple cobalt catalyst system for the efficient and regioselective cyclotrimerisation of alkynes. AB - The intermolecular cyclotrimerisation of terminal and internal alkynes can be catalysed by simple cobalt complexes such as a CoBr2(diimine) under mild reaction conditions when treated with zinc and zinc iodide with high regioselectivity in excellent yields. PMID- 15756341 TI - Selective guanine oxidation by UVB-irradiation in telomeric DNA. AB - The combination of the transient absorption measurement and DNA damage quantification by HPLC clearly demonstrated the preferential excitation of guanine and its decomposition in quadruplex DNA by UVB-irradiation. PMID- 15756342 TI - A simple method for the alpha-oxygenation of aldehydes. AB - A mild, efficient and general method for the chemospecific alpha-oxygenation of aldehydes is described. Treatment of a series of aldehydes with N-tert-butyl-O benzoyl hydroxylamine hydrochloride gives the corresponding alpha-oxygenated carbonyl via a proposed pericyclic rearrangement process. PMID- 15756343 TI - Easy one-pot synthesis of new dppm-type linkers for immobilizations. AB - New synthetic routes to symmetric and unsymmetric dppm-type chelate linkers containing the ethoxysilane group are described and the compounds were characterized also by solid-state NMR spectroscopy and X-ray structures. PMID- 15756344 TI - Heterogeneous assembly of silver(I) and calcium(II) ions accompanying a dimer formation of cyclo(L-Ala-L-Met)3. AB - Heterogeneous assembly of three Ag+ and one Ca2+ ion has been accomplished in a dimeric structure of cyclic hexapeptides, cyclo(l-Ala-l-Met)3. PMID- 15756345 TI - Questions regarding the role of protein phosphorylation in HIV replication and anti-acquired immune deficiency syndrome therapy. PMID- 15756346 TI - Cholesterol. How low should we go? AB - The National Cholesterol Education Program NCEP has recently updated their Adult Treatment Panel (ATP) III guidelines and called for more intensive cholesterol treatment, especially in patients at high risk for coronary heart disease CHD. The message from the updated report is that lower is better for high risk patients, with the NCEP expert panel calling for low-density lipoprotein LDL- cholesterol treatment targets of <100 mg/dL in patients at high risk for CHD. In very high risk patients, however, aggressively lowering LDL-cholesterol to <70 mg/dL is now a therapeutic option for clinicians. Very high risk individuals are those with cardiovascular disease plus diabetes, persistent cigarette smoking, poorly controlled hypertension, or multiple risk factors of the metabolic syndrome, and those who recently had a myocardial infarction MI. Despite the strong clinical evidence and widely publicized treatment guidelines, many hyperlipidemic patients receive inadequate lipid-lowering treatment or leave the hospital after having a MI without a statin. Intensive therapy should be considered for all patients admitted to the hospital for acute coronary syndrome. Achieving very low levels of LDL-cholesterol often requires high doses of a statin or a combination therapy. The coadministration of ezetimibe, a new cholesterol-absorption inhibitor, further reduced LDL-cholesterol by 23% compared with those patients who remained on statin therapy alone. Recent trials with statin therapy are discussed in this review. PMID- 15756347 TI - Wingless signaling pathway family relation to colon cancer. Have we come full circle? AB - Colorectal cancer CRC is one of the most common malignancies worldwide. Advances in molecular techniques have provided deep insight into the molecular pathogenesis, biologic and genetic changes occurring in colon cancer patients. Current theories of malignant transformation postulate that development of colon cancer is related to 2 main pathways; the loss of heterozygosity pathway, which is usually due to a defect in the adenomatous polyposis coli APC gene and microsatellite instability, which is usually due to a defect in mismatch repair MMR genes. This review summarizes the role of the wingless signaling pathway genes including APC and MMR genes in the development of CRC. PMID- 15756348 TI - Al-Aqeel Sewairi syndrome, a new autosomal recessive disorder with multicentric osteolysis, nodulosis and arthropathy. The first genetic defect of matrix metalloproteinase 2 gene. AB - OBJECTIVE: We report a distinctive autosomal recessive multicentric osteolysis in Saudi Arabian families with distal arthropathy of the metacarpal, metatarsal and interphalangeal joints, with ultimate progression to the proximal joints with decreased range of movements and deformities with ankylosis and generalized osteopenia. In addition, they had large, painful to touch palmar and plantar pads. Hirsutism and mild dysmorphic facial features including proptosis, a narrow nasal bridge, bulbous nose and micrognathia. METHODS: Using a genome-wide search for microsatellite markers from 11 members of the family from the Armed Forces Hospital and King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia, localized the disease gene to chromosome 16q12-21. Haplotype analysis with additional markers narrowed the critical region to 1.2cM and identified the matrix metalloproteinase 2 (MMP-2), (gelatinase A, collagenase type IV, EC 3.4, 24,24) gene as a disease candidate at Mount Sinai School of Medicine, New York, United States of America in April 2000. RESULTS: Some affected individuals were homoallelic for a nonsense mutation (TCA>TAA) in codon 244 of exon 5, predicting the replacement of a tyrosine residue by a stop codon in the first fibronectin type II domain (Y244X). Other affected members had a missense mutation in exon 2 arginine 101-histidine (R101H) leading to no MMP-2 enzyme activity in serum or fibroblast or both of affected individuals. In other affected members, a non-pathogenic homoallelic GT transversion resulted in the substitution of an aspartate with a tyrosine residue in codon 210 of exon 4 (D210Y). The MMP-2-null mouse has no developmental defects, but are small, which may reflect genetic redundancy. CONCLUSION: The discovery that deficiency of this well-characterized gelatinase/collagenase results in an inherited form of an osteolytic and arthritic disorder provides an invaluable insights for the understanding of osteolysis and arthritis and is the first genetic evidence that MMP2 deficiency is important in growth and development. PMID- 15756349 TI - Magnetic resonance imaging of pelvic and femoral bones for detection of bone marrow infiltration in patients with non-Hodgkins lymphoma. AB - OBJECTIVE: Detection of the residual bone marrow (BM) involvement is essential in treatment of patients with non-Hodgkin's lymphoma (NHL). Bilateral iliac crest BM biopsies appear to have low sensitivity for BM infiltration, while magnetic resonance imaging (MRI) presents more global view of BM. This study was conducted to determine the advantage of femoral marrow MRI as a non-invasive technique over bilateral iliac crest BM biopsies in detecting BM infiltration before treatment and residual disease after completion of treatment. METHODS: Over a period of 27 months from January 2002 to March 2004, a total of 30 patients with newly diagnosed NHL were included in the study. Magnetic resonance imaging of 26 patients were conducted in King Fahd Hospital of the University, Al-Khobar, Kingdom of Saudi Arabia. Magnetic resonance imaging of 4 patients were referred from different hospitals in the Eastern Province. Twenty-five patients were of B cell type and 5 patients were of T-cell type. Coronal MRI of the pelvis and femoral marrow were obtained by the T1, T2-weighted spin echo sequences and short TI inversion recovery technique. RESULTS: Magnetic resonance images showed BM infiltration in 17 cases (56.7%) before treatment and positive biopsy results were found in 9 cases (30%); all had abnormal MR images. There was a significant difference between both methods in the detection of infiltration (p=0.037). Magnetic resonance imaging showed that 58.8% of cases have scattered pattern, 23.5% were uniform and 17.6% of cases have nodular patterns. Magnetic resonance images after completion of treatment showed residual BM infiltration in 6 out of 17 cases who previously had positive MR images and only one case of them had a positive BM biopsy with a significant difference between both methods and all of them relapsed within 6 months (p=0.034). CONCLUSION: Magnetic resonance images of the pelvis and femoral marrow were superior than BM biopsy on detection of BM infiltration before treatment and residual infiltration after treatment. PMID- 15756350 TI - Expression of p53 oncoprotein and bcl-2 in renal cell carcinoma. AB - OBJECTIVE: The aim of the present study was to investigate the significance of p53 and bcl-2 as prognostic factors among others in renal cell carcinoma patients. METHODS: We evaluated the stages, histological grades, tumor diameters, cellular patterns and the presence of mutant p53 protein and bcl-2 overexpression in 57 cases of renal cell carcinoma (RCC). Kaplan-Meier and log-rank tests estimated the survival function of each parameter. The study was carried out in the Department of Pathology and the Department of Urology, Faculty of Medicine, Dicle University Hospital, Diyarbakir, Turkey, in 2003. RESULTS: The p53 mutation was 35% and bcl-2 overexpression incidence was 89.4% in the RCC cases included in the study. The 5-year disease specific survival rates of mutant p53 positive was 46.6% and p53 negative cases were 83.3%, (p=0.0063). There was no pathological parameter associated in bcl-2, and it has no prognostic significance. CONCLUSION: The tumor stage, grade, diameter and p53 mutations affect the survival of RCC cases. The bcl-2 staining did not play any role to estimate patients at high risk of the disease progression. PMID- 15756351 TI - The diagnostic accuracy of fine needle aspiration cytology versus core needle biopsy for palpable breast lump(s). AB - OBJECTIVE: Breast lump is one of the most common surgical problems in females. Surgical biopsy of palpable breast lump was considered the gold standard for the diagnosis of breast lump(s). Emphasis has been placed on improving method for establishing a definitive diagnosis of breast mass prior to surgery. Our aim was to compare the diagnostic accuracy of fine needle aspiration cytology (FNAC) and core needle biopsy (CNB) in patients with palpable breast masses. METHODS: Prospective randomized controlled clinical trial included 296 Female patients with breast lumps, presented to Al-Thawra Teaching Hospital, Sanaa, Yemen between May 1998 -- May 2002. All the patients submitted either to FNAC or CNB equally. The results were then compared with the final diagnosis by histopathology. RESULTS: The patient age ranged from 15-74 years with a mean of 33.77 +/- 11.91 years. Married patients were 69.3% and the most common presentation was breast lump(s) (88.5%). The mean size of the tumor was 3.47 +/- 1.43 cm in diameter. The FNAC sensitivity was 66.66%, 81.8% specificity, 75.7% accuracy, positive predictive value (PPV) 100% and negative predictive value (NPV) 90%, while in core needle breast biopsy sensitivity was 92.3%, 94.8% specificity, 93.4% accuracy, PPV 100% and NPV 100%. The diagnostic accuracy of CNB was higher than the FNAC, which was statistically significant (p<0.05). CONCLUSION: Both procedures are simple, easy, safe, cheap and reliable, but CNB is more accurate than the FNAC. PMID- 15756352 TI - Carcinoma of unknown primary site. AB - OBJECTIVE: To verify the spectrum of manifestations of carcinoma of unknown primary site, histopathological type and outcome, as well as to identify the prognostic factors for patient's outcome and their survival rate. METHODS: From January 2001 to July 2002, in Basrah Teaching Hospital, Basrah, Iraq, 60 patients (27 males and 33 females), (mean age 58.8 +/- 11.1 years) who had fulfilled the criteria for carcinoma of unknown primary site were studied. RESULTS: Forty-one patients were with one site of metastasis and 19 patients were with multiple site of metastasis. Liver was the most common site of metastasis (in single site group) with 42% of the patients while in the bone with 22% of patients. The mean age of male patients was higher than the female patients (62.1 versus 55 years). Twenty-three female patients were less than 60-years-old (70%), while 10 male patients were less than 60 years (30%). These findings were statistically significant. Adenocarcinoma was the most common histopathological type (63% of cases), while sequamous cell carcinoma was the least (13%). Fifty percent of patients were smokers. Weight loss was the most common presentation (37% of cases). Follow up was made for 48 patients (20 males and 28 females). CONCLUSION: The mean survival was 2.4 +/- 1.8 months. Age > or = 60, smoking, adenocarcinoma type and multiple site of metastasis were bad prognostic factors for the outcome. PMID- 15756353 TI - Creating computer aided 3D model of spleen and kidney based on Visible Human Project. AB - OBJECTIVE: To investigate the efficacy of computer aided 3-dimensional (3D) reconstruction technique on visualization and modeling of gross anatomical structures with an affordable methodology applied on the spleen and kidney. METHODS: From The Visible Human Project Dataset cryosection images, developed by the National Library of Medicine, the spleen and kidney sections were preferred to be used due to their highly distinct contours. The software used for the reconstruction were SurfDriver 3.5.3 for Mac and Cinema 4D XL version 7.1 for Mac OS X. This study was carried out in May 2004 at the Department of Anatomy, Hacettepe University, Ankara, Turkey. RESULTS: As a result of this study, it is determined that these 2 programs could be effectively used both for 3D modeling of the mentioned organs and volumetric analyses on these models. It is also seen that it is possible to hold the physical models of these gross anatomical digital ones with stereolithography technique by means of the data exchange file format provided by the program and present such images as anaglyph. CONCLUSION: SurfDriver 3.5.3 for Mac OS and Cinema 4 DXL version 7.1 for Mac OS X can be used effectively for reconstruction of gross anatomical structures from serial parallel sections with distinct contours such as spleen and kidney and the animation of models. These software constitute a highly effective way of getting volumetric calculations, spatial relations and morphometrical measurements of reconstructed structures. PMID- 15756355 TI - Detecting of Mycoplasma genitalium in male patients with urethritis symptoms in Turkey by polymerase chain reaction. AB - OBJECTIVE: The aim of this study was to investigate the incidence of Mycoplasma genitalium in the urine samples of 63 male patients who had urethritis symptoms. Along with Neisseria gonorrhoeae (N. gonorrhoeae) and Chlamydia trachomatis (C. trachomatis). We also investigated Mycoplasma hominis (M. hominis) and Ureaplasma urealyticum (U. urealyticum), both of which are known to cause urethritis. METHODS: Microorganisms were investigated in urine samples of the patients with polymerase chain reaction. The study was conducted between September 2003 - February 2004 at the Department of Microbiology and Clinical Microbiology Ankara University School of Medicine, Ankara, Turkey. RESULTS: A total of 63 urine samples were analyzed and 6 (9.52%) patients had N. gonorrhoeae, 4 (6.34%) had C. trachomatis, while 4 (6.34%) urines were positive in terms of M. genitalium. Nevertheless, 3 (4.76%) patients had U. urealyticum and 2 (3.17%) patients had M. hominis. One urine sample was positive in terms of both N. gonorrhoeae and U. urealyticum, and another urine sample was positive in terms of both M. hominis and U. urealyticum. The results were compared with the control group and found no statistically significant difference. CONCLUSION: Mycoplasma species are found in normal flora of urogenital system and also as an agent of urogenital infection. In our study, we found low microorganism rates when compared with Europe and America. This difference may be due to the conservative sexual behavior in Turkey. PMID- 15756354 TI - Comparison of antibiotic susceptibility tests, plasmid profiles and restriction enzyme analysis of plasmid DNA of methicillin susceptible and resistant Staphylococcus aureus strains isolated from intensive care units. AB - OBJECTIVE: To differentiate methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) and methicillin-sensitive S. aureus (MSSA) strains and detect the source of epidemic strains and prevent their access to patients. METHODS: All the procedures were carried out in the Department of Microbiology, Medical Faculty Hospital, Dokuz Eylul University, Izmir, Turkey from 1996-1998, and antibiotic susceptibility tests continued in the laboratory of King Fahad Hospital, Al-Baha, Kingdom of Saudi Arabia (KSA), from 2001-2004. A total of 81 S. aureus strains (71 MRSA, 10 MSSA) from Turkey were isolated from different sites of patients in Intensive Care Unit's (ICU's), evaluated by plasmid profile, Restriction Endonuclease Analysis of Plasmids (REAP), and antibiotic sensitivity tests. A total of 117 S. aureus strains (24 MRSA, 93 MSSA) from KSA were isolated from different sites of patients in ICU's, evaluated by antibiotic sensitivity tests as recommended by National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Seventy-one MRSA from Turkey were divided into 13 groups by antibiotic sensitivity tests and into 4 groups by plasmid profiles, in which 3rd and 4th groups subdivided into 2 subgroups, and into 5 groups by REAP. The 1st, 2nd, 3rd and 5th groups were subdivided into 2 subgroups. Ten MSSA were divided into 4 groups by antibiotic sensitivity tests, 3 in plasmid profiles and 2 in REAP tests. Twenty-four MRSA strains from KSA were divided into 9 groups by antibiotic sensitivity tests while 93 MSSA strains were divided into 7 groups. CONCLUSION: In respect to epidemiological survey, plasmids profiles and REAP seems to discriminate more respect to antibiotic sensitivity tests but at the same time neither of them were 100% accurately differential. According to the plasmid profile of the 3rd MSSA (Turkey) group, a multi-drug resistance by antibiotic susceptibility tests were noticed and showed the same plasmid profile in MRSA first subgroup of the 3rd group, but the same groups were different in REAP tests. In order to distinguish the discriminatory power of the strains, where REAP is better than plasmid profile and antibiotic sensitivity tests, we may formulate the statement into the following; REAP > plasmid profile > antibiotic sensitivity tests. For typing and gathering of epidemiological data, it is suggested that all 3 methods should be employed in clinical laboratories as they are cheap, practical and easily interpreted. PMID- 15756356 TI - In-vitro antimicrobial activity of Lawsonia inermis Linn (henna). A pilot study on the Omani henna. AB - OBJECTIVE: To investigate the antimicrobial activity of henna's fresh and dry leaves and seeds obtained from Oman. METHODS: This study was carried out at the College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Sultanate of Oman during the period January-June, 2004. Crude extracts of fresh and dry leaves and seeds were investigated for their antimicrobial activity against 3 standard bacterial strains namely: Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Eleven different bacterial strains were obtained from patients attending the Sultan Qaboos University Hospital, Muscat, Sultanate of Oman. In addition, one Candida albicans (C. albicans) species was used for testing the antifungal activity of the Omani henna sample. RESULTS: All fresh and dry leaves and seeds of the Omani henna demonstrated antibacterial activity against all 3 standard strains and the 11 patients' isolated strains. Henna dry leaves demonstrated the best in-vitro antimicrobial activity and in particular against Shigella sonnei. However, henna fresh and dry seeds failed to show any activity against C. albicans. CONCLUSION: Omani henna does possess, in-vitro antibacterial activity against a wide spectrum of bacterial strains and C. albicans. PMID- 15756357 TI - Biochemical analysis of serum pancreatic amylase and lipase enzymes in patients with type 1 and type 2 diabetes mellitus. AB - OBJECTIVE: To examine the pancreatic exocrine insufficiency in patient with diabetes mellitus by estimating serum pancreatic amylase and lipase enzymes in healthy subjects and in type 1 and type 2 diabetic patients. METHODS: The study was conducted on 20 normal healthy volunteers and 39 diabetic patients referred to Al-Isra Medical Laboratory, Amman, Jordan during the period from April - November 2003 after recording their age and gender. The age of onset of diabetes and the type of treatment were determined and the patients were categorized into type 1 and type 2 diabetics. Blood samples were collected and analyzed for fasting blood sugar (FBS), glycosylated hemoglobin (HbA1C), serum insulin, and serum pancreatic amylase and lipase enzymes. All biochemical tests were carried out in the medical laboratories of Islamic Hospital, Amman, Jordan. All estimates were presented as means +/- SD, and statistical treatment of data were performed using student t-test. RESULTS: The FBS and HbA1C estimates were consistently higher in type 1 and type 2 diabetic patients, while no significant changes were observed in the estimates of serum insulin between the normal and diabetic patients. The reduction in serum pancreatic amylase was recorded in both types of diabetes, which amounted to 71% for type 1 diabetics and 49% for type 2 diabetics. On the other hand, reduction in serum lipase was only detected in type 1 diabetics amounting to 31%. Correlation of the reduction in serum amylase and lipase levels with the duration of disease revealed a remarkable decrease in both enzymes in patients with long-standing disease (76% and 39%) in type 1 diabetic patients. Whereas, patients with very low serum insulin estimates the reduction in serum amylase was 77% while serum lipase level was reduced by 42%. Similarly, the reduction in serum amylase in type 2 diabetes was higher in patients with longer duration of illness (59%) and in patients with low serum insulin value (79%), while reduction in serum lipase was only detected in patients with very low serum insulin (34%). No differences in all measured parameters between males and females were recorded in type 1 and type 2 diabetics. CONCLUSION: Although most of diabetic research has been focused on dyslipidemia as a major risk factor for cardiac, cerebral and renal complications, the present study clearly illustrates an impairment of pancreatic exocrine function in type 1 and type 2 diabetes. We suggest that analysis of serum pancreatic enzymes could be an additional informative parameter for the assessment of chronicity and progress of the illness as well as the response to therapy. PMID- 15756358 TI - Treatment of complicated and uncomplicated hypertension with nifedipine in Palestine. AB - OBJECTIVE: It is generally agreed that short acting calcium channel blockers (CCB) as nifedipine are not safe as the reflex increase in sympathetic activity and thus, their use in hypertension (HTN) is questionable. The objective of this study was to determine the extent of utilization of short acting nifedipine in the treatment of complicated and uncomplicated HTN in one of the major districts in Palestine, Nablus. METHODS: The data for this study were collected from patient's medical files. Data collection was made over a period of 6 months (April - October 2003) in Nablus, Palestine. The sample studied included 876 patients with cardiovascular diseases. Focus was made on the 262 patients with uncomplicated HTN and the 147 patients with both HTN and diabetes mellitus. RESULTS: A total of 247 (28.2%) patients in the study sample were receiving CCB. One third (33.7%) of patients with uncomplicated HTN were receiving CCB mainly as monotherapy with most of them (61%) were receiving the old generation nifedipine as a short acting formulation. Use of nifedipine was also more common among elderly patients above the age of 60 years. CONCLUSION: There was a tendency to use short acting nifedipine as antihypertensive monotherapy among patients with either uncomplicated or complicated HTN. This irrational use needs to be reviewed since it is associated with higher risks. PMID- 15756359 TI - Functional capacity in severe chronic obstructive pulmonary disease. AB - OBJECTIVE: To investigate the functional capacity in patients with severe chronic obstructive pulmonary disease (COPD) and healthy subjects. METHODS: The present study was conducted in the School of Physical Therapy and Rehabilitation, Department of Chest Medicine, Medical School, Hacettepe University, Ankara, Turkey, from June 2002 to July 2003. Forty-five male patients with severe (23 moderately severe, 22 very severe) COPD and 45 healthy controls participated in the study. Pulmonary function tests and 6-minute walk test (6MWT) were performed. The Borg score and heart rate were measured before and after the test. External work of walking was calculated as the product of body weight and walked distance. Functional dyspnea was measured using the modified Medical Research Council (MRC) dyspnea scale. RESULTS: Both distance and external work of 6MWT were significantly lower in severe COPD patients than healthy subjects (p<0.05). No significant differences in distance, external work, and exertional dyspnea were found between moderately severe and very severe COPD patients (p>0.05). Patients with very severe impairment had a significantly lower percentage of predicted 6MWT distance (p<0.05). In these patients, 6MWT distance was correlated significantly with forced expiratory flow in one second (FEV1), MRC score, and Borg score whereas external work of walking correlated with age, FEV1, forced expiratory flow from 25-75%, MRC, and resting Borg score (p<0.05). CONCLUSION: Severe COPD patients had a significantly impaired functional capacity as compared to their healthy counterparts. Patients with FEV1 <35% have a lower percentage of functional capacity. Both distance and external work of 6MWT are influenced by large and small airway function, level of breathlessness reported during daily activity or scored during an exercise test in patients with FEV1 <35%. PMID- 15756360 TI - Patient preference for providers' gender at a primary health care setting in Alexandria, Egypt. AB - OBJECTIVE: The aim of this study is to determine patient's gender preferences in choosing general practitioners and specialists for both general and specific health issues at primary health care centers in Alexandria, Egypt. METHODS: A descriptive cross-sectional study was conducted at Smouha Health Center, Alexandria Governorate, Alexandria, Egypt. The study was conducted between the period January -- March 2004. A pre-designed questionnaire was used. The format included 3 main sections, namely, socio-demographic, beliefs towards provider's gender, and providers' preferences of patients. RESULTS: Both male and female patients preferred a male physician for consultation and clinical management of cases. The highest percentage of male patients (81.4%) and female patients (41.0%) preferred physician of the same gender, and such preference was of personal reasons for males (97.9%) and females (96.6%), while basing on traditions and norms for male (62.9%) and female (63.4%). Significant results of the stepwise logistic regression analysis in relation to socio-demographic factors on provider preference revealed that job and income were the significant determinants of gender preference for the sample under study. CONCLUSION: Patients prefer physicians of the same gender, but in actual practice a male physician is believed to be more competent. PMID- 15756361 TI - Fetal macrosomia. Risk factor and outcome. AB - OBJECTIVE: To determine the risk factors predisposing to fetal macrosomia and assess the maternal and perinatal outcome in these patients. METHODS: This was a retrospective analysis of all macrosomic deliveries in the Department of Obstetrics and Gynecology, Sultan Qaboos University Hospital, Sultanate of Oman, during a 3-year period from January 2001 -- December 2003. The maternal and neonatal records of infants with birth weight of > or =4000 g (n=275) were reviewed. Outcome variables included demographic profile, antenatal risk factors, mode of delivery and maternal and perinatal complications. RESULTS: A total of 7367 deliveries occurred during the study period. The rate of macrosomic deliveries was 3.75% and the rate of deliveries > or =4500 g was 0.48%. The mean birth weight of the study group was 4230 +/- 220 g. Obesity, diabetes, prolonged gestation and postpartum hemorrhage were significantly higher in the study group. The cesarean section rate was 25.8% for the study group compared to the general incidence of 13.1% during the study period (p<0.0001). The incidence of shoulder dystocia was 7.6% compared to the general incidence of 0.48% during the study period (p<0.0001). There were 7 cases of Erb's palsy, all except one recovered without sequelae by 3 months of age. CONCLUSION: Gestational diabetes, maternal obesity, increasing age and parity were the main risk factors for fetal macrosomia. The incidence of shoulder dystocia, birth injuries and neonatal morbidity increased in this group. PMID- 15756362 TI - Magnetic resonance venography of double inferior vena cava. AB - Duplication of the inferior vena cava IVC is the most common anomaly affecting the vena cava. Variations of the IVC are diagnosed in routine dissection studies, in retroperitoneal surgeries, or in radiological studies for various reasons. In this paper, we present a case of double IVC with its magnetic resonance imaging findings. PMID- 15756363 TI - Jaffe-Campanacci syndrome. AB - This case report describes the clinical, radiological and histopathological features of the Jaffe-Campanacci syndrome as seen in a 6-year-old Qatari male patient who was initially misdiagnosed as a case of systemic neurofibromatosis. Our case has all the diagnostic stigmata of Jaffe-Campanacci syndrome as described in the literature and these include cafe au lait macules, skeletal deformities and multiple histologically confirmed non-ossifying fibromas of the long bones. PMID- 15756364 TI - Pheochromocytoma of the organ of Zuckerkandl. Unusually small lesion detected with magnetic resonance imaging. AB - A case report of a 40-year-old female with an unusually small pheochromocytoma of the organ of Zuckerkandl is discussed. The tumor is diagnosed by magnetic resonance imaging MRI examination, which was requested to evaluate her as a potential renal donor. There is a family history of Carney s triad gastric leiomyosarcoma, extra-adrenal pheochromocytoma and pulmonary chondroma. The MRI technique and findings are discussed. PMID- 15756365 TI - Castleman disease presenting as obstructive jaundice. AB - A 48-year-old Saudi male was admitted with features of obstructive jaundice. Endoscopic retrograde cholangiopancreatography showed stricture in distal common bile duct CBD. Computed tomography scan revealed lymphadenopathy along CBD and in porta hepatis. Cholangiocarcinoma, lymphoma or metastatic deposits were suspected but no definite preoperative diagnosis could be established. Laparotomy disclosed lymph node enlargement in porta hepatis and along the CBD and lesser curvature of the stomach. Triple bypass procedure was performed to relieve the obstruction. Pathological examination of the lymph nodes showed Castleman disease of hyaline vascular type. PMID- 15756366 TI - An unusual etiology of epigastric mass. AB - Bezoar is described as presence of indigested or poorly digested material forming a mass in the gastrointestinal lumen. Patients may present with abdominal pain, dyspeptic complaints, gastrointestinal obstruction, perforation or bleeding, as well as, incidental abdominal mass in asymptomatic patients. We report a 30-year old female patient with no history of previous illness who was presented to the emergency ward with complaints of epigastric pain and abdominal bloating. The diagnostic features and treatment of bezoar are presented. PMID- 15756367 TI - Hallucinogenic plant poisoning in children. AB - Datura is a hallucinogenic plant found in urban or rural areas in the Kingdom of Saudi Arabia KSA. It grows wildly in many parts of the country. Its taste and shape makes it unattractive to both man and animals, though deliberate use by young adults for its hallucinogenic effects have been widely reported for the past 30 years. Datura contains 3 main toxic alkaloids: atropine, scopolamine and hyoscamine. Consumption of any part of the plant can result in severe anticholinergic toxicity. Clinical symptoms are those seen in atropine poisoning, particularly mydriasis and hallucinations. Children have a special susceptibility to atropine toxicity; even small amount may produce central nervous system manifestations. Hospitalization is required for agitation and combative behavior although symptomatic treatment is usually sufficient. We report a case of acute Datura stramonium intoxication in a 6-year-old boy from Khamis Mushayt, KSA, who presented with restlessness, hallucinations and mydriasis 8 hours after ingesting the seeds of Datura plant. PMID- 15756368 TI - Abdominal tuberculosis in adolescents. Difficulties in diagnosis. AB - Since the nature of abdominal tuberculosis is mimicking a number of diseases, this may cause delayed diagnosis resulting in evident increased morbidity and mortality. Most of the time, serologic and bacteriologic tools are not enough. We report 3 adolescents with distinct presentations, one mimicking Crohn's disease, one with hepatitis, and the last one with ascites. Terminal ileitis and mesenteric lymphadenitis were found in laparotomy of the first case mimicking Crohn disease. Granulomatous hepatitis was found in the liver biopsy of the second patient, and peritonitis was found by laparoscopy of the third patient. Tuberculosis could be diagnosed merely by histopathologic investigation. All were treated successfully without complication. PMID- 15756369 TI - Subaortic membrane in an adult patient with coronary artery disease. AB - A case of subaortic membrane with coronary artery disease in a 48-year-old man is described. He was referred to our hospital for cardiac murmur, which was discovered on routine clinical examination. He had no significant past medical history apart from dizziness while exercising. Subaortic membrane was totally excised; left internal thoracic artery was anastomosed to left anterior descending artery by end-to-side technique. The postoperative 2-year course of the patient was uneventful. PMID- 15756370 TI - Successful outcome after evacuation of intracranial hematoma following thrombolysis in acute myocardial infarction. AB - Thrombolytic therapy is the modality of choice for the treatment of life threatening thrombosis in various vascular territories and nowadays, is used extensively in setting of acute myocardial infarction. There is, however, the omnipresent danger of serious bleeding inherently associated with the use of all thrombolytics which if it occurs in the brain, can lead to potentially serious neurological impairment and even death. In our report, we describe the successful surgical management of a streptokinase-induced intracranial hemorrhage. Timely neurosurgical intervention is advocated as the optimal approach for this particular side effect of thrombolytic agents. PMID- 15756371 TI - A large giant cell tumor of the sacrum. Advantage of an abdomino-sacral approach. AB - We report a case of giant cell tumor of the sacrum, presenting with sacral pain, swelling, and change of bowel habits. Rectal examination revealed a huge retrorectal mass fixed to the sacrum but not to the wall of the rectum. Abdominal ultrasonography, computed tomography CT scan, and magnetic resonance imaging MRI showed a huge pelvic mass invading the sacrum. Exploration via posterior sacral approach was not successful due to both, extensive bleeding and difficult accessibility. Re-exploration was carried out 2 days later with the patient in lithotomy position. Using abdomino-sacral approach the mass together with part of the sacrum and the whole coccyx were excised. Histopathology reported giant cell tumor of the sacrum with no evidence of mitosis. The patient was symptomless 12 months after surgery and on follow up. PMID- 15756372 TI - An unexpected spontaneous triplet heterotopic pregnancy. AB - A heterotopic pregnancy is the coexistence of one or more ectopic pregnancies and a single or multiple eutopic pregnancies. This potentially fatal condition rarely occurs in spontaneous conception cycles. We report a case of spontaneous triplet heterotopic pregnancy in a multigravida with no known risk factors. Ultrasound imaging was the diagnostic tool used and left salpingectomy via laparotomy was performed in this case report. PMID- 15756373 TI - Whole lung lavage with intermittent double lung ventilation. A modified technique for managing pulmonary alveolar proteinosis. AB - Whole lung lavage is still the most effective treatment for pulmonary alveolar proteinosis. We report a 21-year-old male diagnosed with pulmonary alveolar proteinosis by open lung biopsy and who underwent whole lung lavage with a modified technique. He showed significant improvement in clinical and functional parameters. The technique of intermittent double lung ventilation during lavage procedure keeps the oxygen saturation in acceptable limits in patients at risk for severe hypoxemia and allows the procedure to be completed in a single setting. PMID- 15756374 TI - Cutaneous hyperpigmentation following nonpermanent henna tattoo. AB - Decorative henna skin painting can provoke severe reactions in the skin. We report 3 cases of unusual and persistent hyperpigmentation following nonpermanent henna tattoo, which alerted us to identify an additional side effect. PMID- 15756375 TI - Spontaneous idiopathic chylothorax in adults. PMID- 15756376 TI - A randomized comparison of sulphadoxine-pyrimethamine and combination of sulphadoxine pyrimethamine with chloroquine in the treatment of uncomplicated falciparum malaria in Eastern Sudan. PMID- 15756377 TI - Analysis of documents used in referral system in Wad Medani, Sudan. PMID- 15756378 TI - Uterine prolapse immediately after labor. PMID- 15756379 TI - Spontaneous bacterial peritonitis due to Hafnia alvei in a patient with peritoneal mesothelioma. PMID- 15756380 TI - Antibiotic resistance patterns of Acinetobacter species isolated in King Hussein Medical Center, Jordan. PMID- 15756381 TI - Anti-Toxoplasma gondii antibodies in patients infected with hepatitis B virus. PMID- 15756382 TI - Hypovolemic cardiac arrest after dental extraction. An unexpected high-flow maxillar arteriovenous malformation. PMID- 15756383 TI - Health insurance. A need of time. PMID- 15756384 TI - Critical reading and critique of medical articles. PMID- 15756385 TI - Nosocomial urinary tract infection. PMID- 15756386 TI - Hyperbaric medicine for necrotizing fasciitis. PMID- 15756388 TI - Surgical innovation and diseases of the foregut. PMID- 15756389 TI - The laparoscopic revolution. PMID- 15756390 TI - Recent advances in colorectal surgery. PMID- 15756391 TI - A current assessment of endoluminal approaches to the treatment of gastroesophageal reflux disease. AB - Over the past decade, a number of endoscopic techniques have been developed as alternatives to medical and surgical treatment of gastroesophageal reflux disease (GERD). The driving force was to provide an outpatient transoral, endoscopic procedure effective in controlling reflux in a portion of patients with GERD. Three major technologies emerged, although each use different approaches to augment the barrier function of the lower esophageal sphincter, mechanisms may be similar. These include Endocinch which tightens the gastroesophageal junction via a set of suture plications around the lower esophageal sphincter, Stretta, which delivers radiofrequency energy at the cardia, and Enteryx, which is an inert polymer injected into the muscle layer of the gastroesophageal junction. To date, the underlying mechanism of action of these procedures has not been completely elucidated, although each alters the compliance of the GEJ and thus its ability to respond to a "refluxogenic stress". The target population currently consists of proton pump inhibitor-dependent GERD patients, with little or no hiatal hernia and without severe esophagitis or Barrett's. The Stretta procedure is the only procedure to date to be subjected to a sham-controlled trial. Registries of complications suggest that these techniques are relatively safe, but serious morbidity including rare mortality have been reported. All can be performed on an outpatient basis. Future comparative studies with predetermined end points, validated outcome measures, prolonged follow-up, and complete complication registries are needed to determine the role of endoscopic procedures in the clinical practice of patients with GERD. Evolution of the current technologies will almost certainly occur, and a commonly performed, efficacious endoscopic antireflux procedure is likely to emerge. PMID- 15756392 TI - Initiation and progression of physical activity after laparoscopic and open gastric bypass surgery. AB - This study compared postoperative physical activity participation among patients who underwent laparoscopic (LGBS) or open gastric bypass surgery (OGBS). Postoperative physical activity participation is considered important for achieving optimal weight loss and maintenance after gastric bypass surgery. However, no study has examined the relationship between surgery type and postoperative physical activity. Minimal invasiveness and reduced recovery time associated with LGBS compared with OGBS may permit earlier initiation and faster progression of postsurgical physical activity and potentially contribute to greater long-term adherence rates. Self-reported physical activity participation and aerobic physical activity hours per week at 2-weeks, 3-months, and 6-months postsurgery were assessed among LGBS and OGBS patients (presurgical body mass index of 35 to 70 kg/m(2)) at a university hospital from 1988-2002. Of the 2,235 patients, 531 (24%) and 1704 (76%) underwent LGBS and OGBS, respectively. A greater proportion of LGBS patients reported physical activity participation at each time point compared with OGBS patients (2 week, 76% vs 62%; 3 months, 84% vs 74%; 6 months, 85% vs 76%). Furthermore, LGBS patients reported a significantly greater physical activity duration at 2-weeks postsurgery compared with OGBS patients. A nonsignificant trend toward greater physical activity duration was observed in the LGBS patients at 3 months, whereas 6-month physical activity duration was similar between groups. LGBS, compared with OGBS, may promote earlier onset, progression, and maintenance of physical activity until 6 months postsurgery. Future studies need to prospectively determine whether LGBS, via facilitation of greater engagement in postsurgical physical activity, contributes to more successful weight loss and weight maintenance compared with OGBS. PMID- 15756393 TI - Novel technology and innovations in colorectal surgery: the circular stapler for treatment of hemorrhoids and fibrin glue for treatment of perianal fistulae. AB - The introduction of new techniques and technologies in medical science is both stimulating and controversial. This article is a review of the current status of two such advances. Since its first description, the so-called "stapled hemorrhoidectomy" has been gaining increasing popularity, at first in Asia and Europe, and more recently in the United States. It is obviously a misnomer, since no excision of hemorrhoidal tissue is undertaken in this procedure. It is probably the most significant change in the surgical treatment of hemorrhoids since the introduction of conventional hemorrhoidectomy. Patients routinely experience less postoperative pain and have excellent control of symptoms, with few serious complications in most series. Despite a relatively simple operative technique, the procedure still has specific steps and features that must be followed and mastered to help insure success. The use of fibrin glue for treatment of perianal fistulae has also been a controversial issue, thus it is seldom included in any algorithm as a therapeutic step for fistula-in-ano. The reported success rates of the treatment range from 0% to 100% owing to the heterogeneity of the clinical trials, treatment protocols, patients, etiologies, and types of fistulae. However, the benign nature, simplicity, negligible morbidity, and repeatability of the treatment, potentially makes fibrin glue an attractive first line treatment for perianal fistulae. PMID- 15756394 TI - Endoscopy and surgeons. PMID- 15756395 TI - Ampullary tumors: endoscopic versus operative management. AB - Most ampullary adenomas (80%) are common benign ampullary tumors; however, they can range from mild dysplasia to high-grade dysplasia to invasive carcinoma. They are considered premalignant lesions found in the setting of familial polyposis syndromes or found sporadically, usually manifested by vague abdominal pain, liver enzyme elevation, jaundice, recurrent pancreatitis, or with uncommon symptoms such as gastrointestinal bleeding or duodenal obstruction. Endoscopic retrograde cholangiopancreatography with biopsy is a minimally invasive technique used to visualize these tumors directly and to evaluate their histologic characteristics. Definitive treatment primarily depends on these histologic results. Local resection has a high rate of recurrence (5% to 30%) and requires postoperative endoscopic surveillance, which is the reason it is not considered as a first choice in the management of ampullary tumors. The operative mortality is 10% or less for pancreaticoduodenectomy, a procedure of choice at most experienced centers for frank carcinoma, foci papillary adenocarcinoma in pre excisional biopsies, or high-grade dysplasia ampullary adenomas. Endoscopic interventions for presumed benign ampullary adenomas have resolved symptoms of obstruction, but long-term follow up is necessary to detect early malignant transformation. In summary, the choice of treatment depends on level of surgical skill available, patient tolerance of long-term endoscopic surveillance versus radical surgery, and the presence or absence of coexisting familial adenomatous polyposis. PMID- 15756396 TI - Basic science and the surgeon. PMID- 15756397 TI - Improving the quality of minimally invasive surgery. PMID- 15756398 TI - Surgical education and training. PMID- 15756399 TI - Minimally invasive surgeons of Texas: a new concept in fellowship training. AB - Minimally invasive fellowship training has seen phenomenal growth, with 94 organized fellowship programs in North America. Training experience is varied across programs owing to the differences in surgical practices. Many programs are often weighted to a specific field of advanced laparoscopic surgery, which can limit a fellow's clinical experience. Minimally Invasive Surgeons of Texas (MIST) is a combined organization of four independent private practice and academic MIS fellowships brought together for the common goal of improved fellowship training. PMID- 15756400 TI - The last thing we need is another journal, but... PMID- 15756401 TI - Surgical leadership in the business of surgery. PMID- 15756402 TI - A broader view of marketing: implications for surgeons. AB - The health care sector of the economy is changing. There is unprecedented growth in the health care sector, and competitive forces have a more prominent role. In addition, consumers have become more informed and as a result, more empowered. Patients in the health care sector are no exception to this trend. As patients become more informed, it is imperative that health care providers become more effective at marketing their services. In general, physicians typically have received little training in the field of marketing, which results in potentially limited understanding of the key marketing issues being faced in today's health care environment. We identify and examine several key marketing issues critical to the success of health care providers in today's environment. Further, we offer some managerial recommendations designed to address each of these issues. PMID- 15756403 TI - Comes the revolution ... PMID- 15756404 TI - Harold Hopkins (1918-1995): "let there be light...". PMID- 15756406 TI - [Brainstem injury diagnosed by MRI. An epidemiologic and prognostic reappraisal]. AB - INTRODUCTION AND OBJECTIVES: CT-scan allows emergency surgical evaluation of head injury lesions, but does not offer a comprehensive diagnosis of the resulting brain injuries. Magnetic Resonance Imaging (MRI) can complete the evaluation of head injury, particularly in the brain stem. We attempted to estimate the frequency of traumatic primary brain stem injuries MATERIAL AND METHODS: Thirty patients with moderate or severe head injury (GCS < or = 13) underwent a MRI study during the first two weeks after trauma. In order to exclude old patients with previous ischemic lesions unrelated to the head trauma, only young patients (16-40 years-old) were included. Patients with cranial surgery were also eliminated from the study. Based on previous studies, the FLAIR (8000/120/T. Inversion 2200mseg) sequence was selected. RESULTS: Brain stem injuries were detected in 26.6% of the patients; this was confirmed by two independent radiologists. Six patients had hyperintense lesions compatible with diffuse axonal damage, and two others showed hemorrhagic lesions. These findings were directly related to a specific neurological deficit in four patients; while in the remaining, unspecific consciousness disturbances were noted. CONCLUSIONS: We believe that the FLAIR sequence demonstrate a type of traumatic brain stem injury (probably corresponding to diffuse axonal injury) that is more frequent and less severe in terms of prognosis than those classically described in previous CT scan studies. PMID- 15756405 TI - [Comparison of the effectiveness of pentobarbital and thiopental in patients with refractory intracranial hypertension. Preliminary report of 20 patients]. AB - OBJECTIVE: To assess the effectiveness of pentobarbital and thiopental to control raised intracranial pressure (ICP), refractory to first level measures, in patients with severe traumatic brain injury. MATERIAL AND METHODS: Prospective, randomized study to compare the effectiveness between two treatments: pentobarbital and thiopental. The patients will be selected from those admitted to the Intensive Care Unit with a severe traumatic brain injury (postresuscitation Glasgow Coma Scale equal or less than 8 points) and raised ICP (ICP>20 mmHg) refractory to first level measures according to the Brain Trauma Foundation guidelines. The adverse effects of both treatments were also collected. RESULTS: We present the results of the first 20 patients included. Ten received pentobarbital and the other ten thiopental. There were no statistically significance differences in patients'characteristics (age, sex, severity of the trauma at admission and comorbilities). There were no differences between both groups neither in the Glasgow Coma Scale at admission (thiopental six points; pentobarbital seven points; P=0.26) nor in the admission Cranial Tomography, according to the Traumatic Coma Data Bank Classification. Thiopental treatment controlled raised ICP in five cases and pentobarbital in two cases (P=0.16). Five patients in the thiopental group died and eight in the pentobarbital group (P=0.16). There were no statistically differences between both groups regarding to the presence of hypotension (P=1) or infectious complications. CONCLUSIONS: These preliminary results indicate that thiopental could be more effective than pentobarbital in patients with refractory intracranial hypertension. These results support previous experimental findings that show that both treatments are not equal and justify to continue this study. PMID- 15756407 TI - [Hemifacial spasm as clinical presentation of intracranial meningiomas. Report of three cases and review of the literature]. AB - Hemifacial spasm (HFS) is a clinical entity consisting of brief clonic jerking movements of the facial musculature, beginning in the orbicularis oculi with downward spreading to other facial muscles. Apart from vascular loop compression at the root exit zone of the facial nerve, other causes of HFS are rare. It is exceptional as a form of presentation of intracranial meningiomas We report three cases of patients with meningiomas who presented with HFS, either as an isolated sign or associated with symptoms of rise intracranial pressure or focal deficit. We review the literature and discuss the possible physiopathological mechanisms responsible for this association. PMID- 15756408 TI - [Transnasal-transsphenoidal endoscopic surgery of the sellar region]. AB - Transnasal transsphenoidal endoscopic approach to the sella turcica has recently become an option to the pituitary surgery which could predominate in a near future. OBJECTIVES: To evaluate through the analysis of our results whether it is possible to perform in our Hospital the endoscopic transnasal transsphenoidal approach to the sella with results at least as good as those we obtain with the microscopic sublabial transseptal transsphenoidal approach. MATERIAL AND METHODS: As the first stage of a three stage randomized trial, this is a prospective study of 10 patients operated on, using transnasal transsphenoidal endoscopic approach between august 2002 and August 2003 at the department of neurosurgery of "Hermanos Ameijeiras Hospital". RESULTS: Six patients had pituitary macroadenomas and four had cerebrospinal fluid leak through the sellar floor. In five of six macroadenomas total tumor resection was obtained and in the other one a subtotal resection was performed. Two of four patients with cerebrospinal fluid leak could be cured while the other two patients do not. Complications were present in 30% of cases, but in only 1 (10%) it reached the postoperatory period. CONCLUSIONS: Transnasal transsphenoidal endoscopic approach to the sella can be done save enough in our Hospital to justify the start of the second stage of one randomized trial to determine if the endoscopic technique is better than the usual transsphenoidal approach. PMID- 15756409 TI - [Chiari I malformation: postsurgical evolution after 2 years. Report of ten cases]. AB - OBJECTIVE To describe the clinical and radiological evolution of ten patients with Chiari I malformation (CIM), treated surgically. METHOD: Ten patients with Chiari I malformation underwent suboccipital craniectomy, laminectomy of C1 and dural patch grafting. All of them were followed for at least two years. A magnetic resonance imaging was performed 6 months after surgery. Clinical preoperative evolution and time of installation, results of MRI and clinical postoperative evolution were correlated. RESULTS: In postoperative clinical evolution, we noted an important improvement in sensitive function, decrease of the syringohydromielia and partial improvement of cerebellar functions. Improvement of the motor alteration was less marked. During the two years after treatment the muscular atrophy did not change. CONCLUSIONS: Chiari I malformation is a congenital disease with many clinical manifestations which usually increase with time. A suitable diagnostic studies as well as an appropriate treatment are needed to improve the neurological results. PMID- 15756411 TI - [Cocaine abuse as precipitating factor of tumoral bleeding in an oligoastrocitoma]. AB - Cocaine abuse has been associated with a variety of intracranial haemorrhagic disorders, such as intracerebral, subdural or subarachnoidal haemorrhage. Frequently, these patients harbour underlying vascular malformations, like cerebral aneurysms or arteriovenous malformations (AVM). To the best of our knowledge only two cases of tumoral haemorrhage induced by cocaine abuse have been previously reported. We describe a new case of intracerebral haemorrhage after cocaine inhalation, in which both the preoperative imaging studies and the pathological examination showed a brain tumour as the origin of the haemorrhage. We think that cocaine abuse may be considered as a new precipitating factor in intratumoral haemorrhage. PMID- 15756410 TI - [Overview of the recent clinical trials in severe head injury and analysis of their therapeutic failure]. AB - During the past "Decade of the Brain" several neuroprotective agents have been tested in phase III clinical trials for severe head injury (SHI) but unfortunately none of them significantly improved the outcome of these patients. In contrast to the success achieved by these drugs in animal laboratory studies, the results in terms of neuroprotection in the clinical setting have been disappointing. This paper has been divided in three parts: in the first one, we summarize the pathophysiological mechanisms related to SHI, targeted by the neuroprotective agents. In the second part we review the main clinical trials carried out for SHI to date, and in the third one, we analyze the possible reasons that explain why these agents have failed to show efficacy. PMID- 15756412 TI - Pott's puffy tumor of the vertex years after trauma in a diabetic patient: case report. AB - Pott's puffy tumor is a rare clinical entity characterized by subperiosteal abscess associated with osteomyelitis. It is usually seen as a complication of frontal sinusitis or trauma. This is the unique report of a Pott's puffy tumor located over the vertex of a type 1 diabetic patient with an unusual latency of 14 years following injury. A 27-year old man presented with pain and a soft swelling on his vertex. Magnetic resonance imaging demonstrated subperiosteal abscess in the vertex region associated with dural thickening and perisinusal irregularities of epidural space. Further history revealed that he had a trauma to the same location when he was 13 years old. Considering possible complications due to proximity of the lesion to the sagittal sinus, we retrained from aggressive surgical interventions. We treated our patient with a simple surgical abscess drainage followed by prolonged use of antibiotics and achieved complete therapy. The cellular and humoral elements of the immune system may be disrupted in diabetic patients resulting in such atypical courses and complications of infections. We want to emphasize both importance of the prompt diagnosis of Pott's puffy tumor as intracranial invasion may cause severe neurologic problems, and importance of a surgical intervention tailored for the individual lesion. PMID- 15756413 TI - Hemangioblastoma of the lateral ventricle: case report and review of the literature. AB - We report a unique case of hemangioblastoma of the lateral ventricle in a 73 year old man with cognitive deficits and fluent dysphasia. He harboured an intraventricular tumor, placed at the trigone of the left lateral ventricle. The tumor was successfully excised, by means of a temporal craniotomy. The patient became mute in the immediate postoperative with restoration of speech within a few days. The literature has been reviewed and only three other similar cases have been reported. Discovery of lesions in such unusual location should raise a high degree of suspicion for von Hippel-Lindau disease. PMID- 15756414 TI - [Idiopathic hypertrophic pachymeningitis of the cavernous sinus mimicking meningioma with complete occlusion of the carotid artery]. AB - Idiopathic hypertrophic pachymeningitis (IHPM) is a rare inflammatory entity characterized by the idiopathic thickening of the dura mater. An increasing number of cases have been reported involving the cavernoss sinus. We present the case of a patient with IHPM who was diagnosed of meningioma and showed a complete occlusion of the intracavernous carotid artery. This 42 years old woman was sent to our hospital to be treated radiosurgicaly with the diagnosis of meningioma. The diagnosis was based on MRI findings and on the temporal resistance to corticoid treatment. A complete study of the patient demonstrated the inflammatory nature of the lesion, which improved after long corticosteroid treatment. The differential diagnosis between meningioma and IHPM may be complex because of the similarities in image findings and the temporal resistance to corticosteroids shown by IHPM, thus we emphasize the usefulness of the SPECT to differentiate between inflammation and tumor in this clinical setting. In addition, our patient showed a total occlusion of the intracavernous carotid artery and, as far as we know, this is the first case with such a finding reported in the literature. PMID- 15756415 TI - [Intracranial trigeminal schwannoma with extension to infratemporal fossa, parapharyngeal space, orbit, maxillary sinus and nasal fossa]. AB - Schwannomas reach 8 to 10% of all intracranial tumors. Most originate at the vestibular root of VIII cranial nerve, but trigeminal tumors are infrequent. We present the case of a patient admitted at the National Institute of Neurology and Neurosurgery "Manuel Velasco Suarez" (Mexico City) with a mass occupying the infratemporal fossa with involvement of nearby structures. Schwannomas with extension to the infratemporal fossa are rare. We review the anatomy of this region, the surgical approaches, which have been used and propose a different approach. PMID- 15756416 TI - Expression of human papillomavirus type 16 L1 protein in transgenic tobacco plants. AB - To develop a plant expression system for the production of the human papillomavirus type 16 (HPV16) vaccine, we investigated whether the HPV16 L1 protein can be expressed in tobacco plants and whether it can be used as the cheapest form of edible vaccine. The HPV16 L1 coding sequence was amplified by PCR using specific primers from the plasmid pGEM-T-HPV16 containing the template sequence, and subcloned into the intermediate vector pUCmT and binary vector pBI121 consecutively to obtain the plant expression plasmid pBI-L1. The T-DNA regions of the pBI-L1 binary vector contained the constitutive Cauliflower mosaic virus (CaMV) 35S promoter and the neomycin phosphotransferase npt II gene, which allowed the selection of transformed plants using kanamycin. The tobacco plants were transformed by co-cultivating them, using the leaf disc method, with Agrobacterium tumefaciens LBA4404, which harbored the plant expression plasmid. The regenerated transgenic tobacco plants were selected using kanamycin, and confirmed by PCR. The results of the Southern blot assay also showed that the HPV16 L1 gene was integrated stably into the genome of the transformed tobacco plants. The Western blot analysis showed that the transformed tobacco leaves could express the HPV16 L1 protein. Furthermore, it was demonstrated by ELISA assay that the expressed protein accounted for 0.034%-0.076% of the total soluble leaf protein, was able to form 55 nm virus-like particles compatible with HPV virus-like particle (VLP), and induced mouse erythrocyte hemagglutination in vitro. The present results indicate that the HPV16 L1 protein can be expressed in transgenic tobacco plants and the expressed protein possesses the natural features of the HPV16 L1 protein, implying that the HPV16 L1 transgenic plants can be potentially used as an edible vaccine. PMID- 15756417 TI - Molecular cloning of MSRG-11 gene related to apoptosis of mouse spermatogenic cells. AB - Beginning with a new contig of the expressed sequence tags (Mm.63892) obtained by comparing testis libraries with other tissue and cell line libraries using the digital differential display program, we cloned a new gene which is related to the apoptosis of mouse spermatogenic cells using the Genscan program and polymerase chain reaction (PCR) technology. The sequence data have been submitted to the GenBank database under accession number AY747687. The full cDNA length is 1074 bp, and the gene with 7 exons and 6 introns is located in mouse chromosome 1 H5. The protein is recognized as a new member of calmodulin (CaM) binding protein family because the sequence contains three short calmodulin-binding motifs containing conserved Ile and Gln residues (IQ motif) and is considered to play a critical role in interactions of IQ motif-containing proteins with CaM proteins. The putative protein encoded by this gene has 192 amino acid residues with a theoretical molecular mass of 23.7 kDa and a calculated isoelectric point of 9.71. The sequence shares no significant homology with any known protein in databases. RT-PCR and Northern blot analyses revealed that 1.3 kb MSRG-11 transcript was strongly expressed in adult mouse testis but weakly expressed in the spleen and thymus. The MSRG-11 gene was expressed at various levels, faintly at two weeks postpartum and strongly from three weeks postpartum in adult testes. The green fluorescence produced by pEGFP-C2/MSRG-11 was detected in the cytoplasm of COS7 cells 24 h post-transfection. The pcDNA3.1(?-)/MSRG-11 plasmid was constructed and introduced into COS7 cells using Lipofectamine 2000 transfection reagent (Invitrogen, Carlsbad, USA). MSRG-11 can accelerate COS7 cell apoptosis, which suggests that this gene may play an important role in the development of mouse testes and is a candidate gene of testis-specific apoptosis. Based on these observations, it was considered that we cloned a new gene which probably accelerates spermatogenetic cell apoptosis in mouse. PMID- 15756418 TI - Protein secondary structure prediction using dynamic programming. AB - In the present paper, we describe how a directed graph was constructed and then searched for the optimum path using a dynamic programming approach, based on the secondary structure propensity of the protein short sequence derived from a training data set. The protein secondary structure was thus predicted in this way. The average three-state accuracy of the algorithm used was 76.70%. PMID- 15756419 TI - Alterations of lymphoid enhancer factor-1 isoform expression in solid tumors and acute leukemias. AB - Two major transcripts of lymphoid enhancer factor-1 (LEF-1) have been described. The long isoform with b-catenin binding domain functions as a transcriptional enhancer factor. The short isoform derives from an intronic promoter and exhibits dominant negative activity. Recently, alterations of LEF-1 isoforms distribution have been described in colon cancer. In the current study we employed a quantitative real-time reverse transcription PCR method (TaqMan) to analyze expression of LEF-1 isoforms in a large cohort of human tumor (n = 304) and tumor free control samples (n = 56). The highest expression level of LEF-1 was found in carcinoma samples whereas brain cancer samples expressed little. Expression of LEF-1 was different in distinct cancer types. For example, the mRNA level of LEF 1 was lower in testicular tumor samples compared with tumor-free control samples. Besides epithelial cancers, significant LEF-1 expression was also found in hematopoietic cells. In hematological malignancies, overall LEF-1 level was higher in lymphocytic leukemias compared with myeloid leukemias and normal hematopoiesis. However, acute myeloid leukemia and acute lymphocytic leukemia showed a significantly increased fraction of the oncogenic LEF-1 compared with chronic lymphocytic leukemia and chronic myeloid leukemia. Taken together, these data suggest that LEF-1 is abundantly expressed in human tumors and the ratio of the oncogenic and the dominant negative short isoform altered not only in carcinomas but also in leukemia. PMID- 15756421 TI - Isolation, sequence analysis and expression profile of a novel swine gene differentially expressed in the Longissimus dorsi muscle tissues from LandracexLarge white cross-combination. AB - The mRNA differential display technique was performed to investigate the differences in gene expression in the Longissimus dorsi muscle tissues from LandracexLarge White cross-combination. One novel gene that was differentially expressed was identified using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and its complete cDNA sequence was obtained using the rapid amplification of cDNA ends (RACE) method. The nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence prediction analysis revealed that the open reading frame of this gene encodes a protein of 260 amino acids that contains the putative conserved domain of the carbonic anhydrase, and this protein has high homology with the carbonic anhydrase III (CA III) of four species mouse (91%), horse (91%), rat (89%) and human (86%)-so that it can be defined as swine carbonic anhydrase III. The phylogenetic tree analysis revealed that the swine CA-III has a closer genetic relationship with the horse CA-III than with those of mouse, rat and human. The tissue expression analysis indicated that the swine CA-III gene is generally expressed in most tissues. Our experiment is the first to establish the primary foundation for further research on the swine CA-III gene. PMID- 15756420 TI - Critical role of Cys168 in noggin protein's biological function. AB - Previous studies have indicated that noggin exerts its neural inducing effect by binding and antagonizing bone morphogenetic protein 4 (BMP4). In order to further clarify the relationship between the structure and the function of noggin, and elucidate the possible mechanism responsible for noggin-BMP4 interaction, we generated three noggin mutants, C168S, C174S and C197S, by using a site-directed mutagenesis method. Ectopic expression of wild-type (WT) noggin, C174S or C197S, in Xenopus animal caps (ACs) by mRNA injection converted the explants (prospective ectoderm) into neural tissue, as indicated by the neural-like morphology and expression of the neural cell adhesion molecule (NCAM) in the ACs. In contrast, ACs expressing C168S suffered an epidermal fate similar to the control caps. Similarly, among the three mutants, only C168S lost the dorsalizing function. These studies highlight the critical role played by Cys168 in noggin's biological activities. It probably participates in the formation of an intermolecular disulfide bridge. PMID- 15756422 TI - Synthesis of reassortant infectious bursal disease virus in chickens injected directly with infectious clones from different virus strains. AB - The infectious bursal disease virus (IBDV), a member of the Birnaviridae family, containing a bisegmented double-stranded RNA genome, encodes four structural viral proteins, VP1, VP2, VP3, and VP4, as well as a non-structural protein, VP5. In the present paper, the segment A from two IBDV strains, field isolate ZJ2000 and attenuated strain HZ2, were inserted into one NaeI site by site-directed silent mutagenesis and subcloned into the eukaryotic expression plasmid pCI under the control of the human cytomegalovirus (hCMV) immediate early enhancer and promoter to construct the recombinant plasmids pCI-AKZJ2000 and pCI-AKHZ2, respectively. Each of the two recombinants was combined with another recombinant pCI plasmid containing the marked segment B of strain HZ2 (pCI-mB), and injected intramuscularly into non-immunized chickens. Two chimeric IBDV strains were recovered from the chickens. Two out of eight chickens in each of two groups showed the bursal histopathological change. The reassortant virus derived from pCI-AKZJ2000/pCI-mB can infect chicken embryos and shows relatively low virulence. We have developed a novel virus reverse genetic approach for the study of IBDV. The results also form the basis for investigating the role of VP1 in viral replication and pathogenecity. PMID- 15756423 TI - Co-expression and immunity of Legionella pneumophila mip gene and immunoadjuvant ctxB gene. AB - The mip gene of Legionella pneumophila and the ctxB gene of Vibrio cholerae were amplified by PCR respectively. The amplified cDNA was ligated to the pcDNA3.1(+) vector. The recombinant plasmids pcDNA3.1-mip and pcDNA3.1-ctxB were identified by restriction analysis and PCR, and further confirmed by sequencing analysis. NIH3T3 cells were transfected with pcDNA3.1-mip and pcDNA3.1-ctxB according to the Lipofection method. Transient and stable products of the co-expression of the mip gene and ctxB gene were detected by immunofluorescence and Western blotting. The results showed that NIH3T3 cells were successfully transfected, and that the transiently and stably co-expressed products can be detected in the transfected cells. To detect the humoral and cellular immune response in immunized mice induced by the co-mmunization of the mip and ctxB genes, female BALB/c mice were immunized intramuscularly with pcDNA3.1-mip and pcDNA3.1-ctxB. The results showed that the specific antibody titer and the cytotoxic T-lymphocyte response for pcDNA3.1-mip immunization and co-immunization were increased compared with that of pcDNA3.1(+) immunization. Furthermore, the specific antibody titer and cytotoxic T-lymphocyte response for co-immunization were increased compared with that of pcDNA3.1-mip immunization. Statistical analysis using one-way analysis of variance (ANOVA) showed that there was a significant difference between the groups (P<0.01). The results indicated that the ctxB gene enhanced the humoral and cellular immune response to the mip gene immunization. These findings provide experimental evidence to support the development of the L. pneumophila DNA vaccine. PMID- 15756424 TI - Application of phage-displayed single chain antibodies in western blot. AB - A phage display single chain fragment variable library constructed on pIII protein of M13 filamentous phage was screened using B-lymphocyte stimulator and FP248 as selective molecules. After four rounds of panning, there was a remarkable enrichment in the titer of bound phages. Twenty phage clones were selected from the last round and screened by means of phage-ELISA. With the antibody phages as primary antibodies in Western blot, we developed a method for detecting the specific antigen. The dilutions of antibody phages depend on the affinity between antibody-displayed phage particles and antigens. PMID- 15756425 TI - In vitro construction of effective M1GS ribozymes targeting HCMV UL54 RNA segments. AB - Seven sequence-specific ribozymes (M1GS RNAs) derived in vitro from the catalytic RNA subunit of Escherichia coli RNase P and targeting the mRNAs transcribed by the UL54 gene encoding the DNA polymerase of human cytomegalovirus were screened from 11 ribozymes that were designed based on four rules: (1) the NCCA-3' terminal must be unpaired with the substrate; (2) the guide sequence (GS) must be at least 12 nt in length; (3) the eighth nucleotide must be U, counting from the site -1; and (4) around the cleavage site, the sites -1/+1/+2 must be U/G/C or C/G/C. Further investigation of the factors affecting the cleavage effect and the optimal ratio for M1GS/substrate was carried out. It was determined that the optimal ratio for M1GS/substrate was 2:1 and too much M1GS led to substrate degrading. As indicated above, several M1GS that cleaved HCMV UL54 RNA segments in vitro were successfully designed and constructed. Our studies support the use of ribozyme M1GS as antisense molecules to silence HCMV mRNA in vitro, and using the selection procedure as a general approach for the engineering of RNase P ribozymes. PMID- 15756426 TI - Aspirin, ibuprofen, and other non-steroidal anti-inflammatory drugs in cancer prevention: a critical review of non-selective COX-2 blockade (review). AB - We comprehensively reviewed the published scientific literature on non-steroidal anti-inflammatory drugs (NSAIDs) and cancer and evaluated results based upon epidemiologic criteria of judgment: consistency of results, strength of association, dose response, molecular specificity, and biological plausibility. Sufficient data from 91 epidemiologic studies were available to examine the dose response of relative risk and level of NSAID intake for ten human malignancies. Dose response curves were fitted by exponential regression. Results showed a significant exponential decline in the risk with increasing intake of NSAIDs (primarily aspirin or ibuprofen) for 7-10 malignancies including the four major types: colon, breast, lung, and prostate cancer. Daily intake of NSAIDs, primarily aspirin, produced risk reductions of 63% for colon, 39% for breast, 36% for lung, and 39% for prostate cancer. Significant risk reductions were also observed for esophageal (73%), stomach (62%), and ovarian cancer (47%). NSAID effects became apparent after five or more years of use and were stronger with longer duration. Observed protective effects were also consistently stronger for gastrointestinal malignancies (esophagus, stomach, and colon). Results for pancreatic, urinary bladder, and renal cancer were inconsistent. Initial epidemiologic studies of malignant melanoma, Hodgkin's disease, and adult leukemia also found that NSAIDs are protective. A few studies suggest that ibuprofen has stronger anticancer effects than aspirin, particularly against breast and lung cancer. Overexpression of cyclooxygenase-2 (COX-2) and increased prostaglandin biosynthesis correlates with carcinogenesis and metastasis at most anatomic sites. Preclinical investigations provide consistent evidence that both selective and non-selective NSAIDs effectively inhibit chemically-induced carcinogenesis of epithelial tumors. This review provides compelling and converging evidence that regular intake of NSAIDs that non-selectively block COX 2 protects against the development of many types of cancer. PMID- 15756427 TI - Rat soluble tumor-associated antigens inhibit chemically-induced mammary tumorigenesis in syngeneic rats. AB - This study examined whether soluble 66 and 51 kDa tumor-associated antigens (sTAA), isolated from the serum of rats with mammary cancer, possess specific suppressive effects on chemically-induced mammary tumorigenesis in syngeneic counterparts. Dimethylbenzanthracene (DMBA, 10 mg/rat, two administrations) was used to induce mammary tumors in 8-week-old Sprague Dawley rats. After the appearance of numerous tumors, preparations of sTAA (50 to 60 microg/rat in 0.5 ml sterile PBS) obtained from breast cancer patients (heterologous sTAA) or from syngeneic mammary tumor-bearing rats (syngeneic sTAA) were administered weekly for 12 weeks. The following groups of mammary tumor-bearing rats were studied: groups 1 and 3, control rats treated with saline; group 2, rats treated with heterologous sTAA; and group 4, rats treated with syngeneic sTAA. The experiment was terminated when tumors in 50% of the rats became ulcerous. The treatment with both types of sTAA significantly decreased, compared to controls and initial values, the yield and total area of the tumors. We conclude that syngeneic sTAA have tumor-suppressive properties, which are very similar to those in heterologous sTAA. PMID- 15756428 TI - Pharmacokinetics and adverse reactions of a new liposomal cisplatin (Lipoplatin): phase I study. AB - Lipoplatin, a new liposomal cisplatin formulation, is formed from cisplatin and liposomes composed of dipalmitoyl phosphatidyl glycerol (DPPG), soy phosphatidyl choline (SPC-3), cholesterol and methoxy-polyethylene glycol-distearoyl phosphatidylethanolamine (mPEG2000-DSPE). Following intravenous infusion, the nanoparticles (110 nm) are distributed into tissues and concentrate preferentially at tumor sites supposedly via extravasation through the leaky tumor vasculature. This study was designed to investigate the pharmacokinetics and the toxicity of this new liposomal cisplatin in patients with pretreated advanced malignant tumors. The drug was infused for 8 h every 14 days at escalating doses. Twenty-seven patients were included and 3-5 patients were selected for each dosage level; levels started at 25 mg/m2 and were increased by 25 to 125 mg/m2. Three patients were also treated at higher dose levels, one each at 200, 250 and 300 mg/m2. Blood was taken at certain time intervals in order to estimate total platinum plasma levels. At level 5 (125 mg/m2), grades 1 and 2 GI tract and hematological toxicities were detected. No nephrotoxicity was observed. Seven additional patients were added at the 4th level (100 mg/m2) for further pharmacokinetic evaluation. Measurement of platinum levels in the plasma of patients as a function of time showed that a maximum platinum level is attained at 6-8 h. The half-life of Lipoplatin was 60-117 h depending on the dose. Urine excretion reached about 40% of the infused dose in 3 days. The data demonstrate that Lipoplatin up to a dose of 125 mg/m2 every 14 days has no nephrotoxicity and it lacks the serious side effects of cisplatin. PMID- 15756429 TI - Expression of thymidine phosphorylase correlates with microvessel density in prostate cancer. AB - Angiogenesis in the growth and development of prostate cancer was the focus of this study. Various angiogenic factors and their clinicopathologic correlations with the progression of prostate cancer have been examined. Thymidine phosphorylase is identical to platelet-derived endothelial cell growth factor (TP/PD-ECGF) and has angiogenic activity. We investigated the expression of TP/PD ECGF in prostate cancer and its association with angiogenesis or clinicopathologic findings in 81 cases with prostate cancer. Western blot analysis using a specific monoclonal antibody 654-1 revealed the existence of a 55 kDa TP/PD-ECGF protein in human prostate cancer tissue. Cancer tissue showed low-positive immunostaining in 32 cases (39.5%) and high positivity in 49 cases (60.5%). This protein expression indicated a statistically significant association with microvessel density (low vs. high TP/PD-ECGF expression group: mean +/- SD, 37.3+/-27.0 vs. 53.1+/-28.0 microvessels in three fields, p<0.05). No correlation was found between the expression of TP/PD-ECGF and nuclear grade, glandular differentiation, clinical stage or overall survival rate. TP/PD-ECGF may play an important role in tumor angiogenesis in prostate cancer tissues. Although the expression of TP/PD-ECGF was not correlated with clinical outcome in patients with prostate cancer, there remains the possibility that TP/PD-ECGF may support or modify the tumor growth through angiogenesis in cooperation with other factors. PMID- 15756430 TI - Relationship between peroxisome proliferator-activated receptor-gamma expression and differentiation of human esophageal squamous cell carcinoma. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, is involved in suppressing the growth of several tumors. We showed that PPAR-gamma is expressed in Barrett's adenocarcinoma cell lines and inhibited the growth of these lines through the induction of G1 cell cycle arrest and apoptosis. We examined PPAR-gamma expression in human esophageal squamous cell carcinoma (SCC) in vitro and in vivo and investigated whether PPAR-gamma ligands affect the proliferation and apoptosis of human SCC cell lines. Biopsy specimens (n=46) obtained from human SCC of the esophagus were stained using a monoclonal antibody against human PPAR gamma. We assessed the effects of PPAR-gamma ligands on the growth of SCC cells by adding 15-deoxy prostaglandin J2 (15d-PGJ2), or troglitazone to six human esophageal SCC cell lines (TE-1, TE-2, TE-3, TE-5, TE-8, and TE-9). Immunohistochemical staining showed that 34 of 46 (73.9%) SCC of the esophagus expressed PPAR-gamma. All SCC cell lines expressed PPAR-gamma mRNA and protein, especially when poorly differentiated (TE-2, TE-5, and TE-9). The PPAR-gamma ligands significantly and dose-dependently inhibited the proliferation of SCC lines, except for well-differentiated TE-1 and TE-3. Apoptosis was induced by 15d PGJ2 (10 microM) in all tested SCC lines except TE-1, whereas troglitazone (50 microM) was marginally effective in only the TE-2 and TE-3 cell lines. The present findings suggest that PPAR-gamma could be a therapeutic target for treating squamous cell carcinoma of the esophagus, possibly through the induction of apoptosis. PMID- 15756431 TI - Changes of histological and biological features by serial passages in a human adenoid cystic carcinoma line transplantable in nude mice. AB - The basic histologic patterns of adenoid cystic carcinoma (ACC) are classified into three types (tubular, cribriform and solid), but clinical significance of the histological type is unclear. We have successfully established a human tumor line derived from ACC that is serially transplantable in nude mice. This tumor showed an increased growth rate as the passage levels proceeded, and the histological type was changed from a cribriform pattern in the initial stage to a solid one. In this study, we investigated the relationship between histological type and biological characteristics by analyzing the serially transplantable ACC tumor model. As a result, the tumor growth rate at the 15th passage level was increased approximately 5-fold compared with that at the initial passage level. In the histological type, approximately 30% of the cribriform pattern in the initial level was changed to a solid one at the 15th passage level, and the PCNA labeling index was elevated 4-fold. Concomitant with this, expression of Ki-67, p53 and bcl-2 proteins was increased, and apoptotic cells were decreased as demonstrated by the TUNEL method. From these findings, it was suggested that cell proliferation and histological change of this ACC tumor are related to the inhibition of apoptosis. This tumor line would provide a useful model for investigating the biological behavior of ACC. PMID- 15756432 TI - Expression of cell adhesion molecules and chemokine receptors: angioinvasiveness in nasal NK/T-cell lymphoma. AB - Sinonasal natural killer (NK)/T-cell lymphoma (NKTCL) is closely associated with Epstein-Barr virus (EBV) infection and expresses latent membrane protein (LMP)-1 and EB nuclear antigen (EBNA)-1, i.e., latency II of EBV infection. Angioinvasion by neoplastic cells is a characteristic feature of NKTCL, but its mechanism is unknown. To elucidate the molecular mechanism of angio-invasiveness in NKTCL, expression of cell adhesion molecules and chemokine receptors at mRNA and protein levels was examined using real-time PCR and immunohistochemistry in 17 NKTCL together with 10 diffuse large B-cell lymphoma (DLBL) and 9 non-neoplastic nasal mucosa as controls. EBV DNA was detected in 14 of 16 NKTCL examined, and 7 of these 14 expressed LMP-1. mRNA expression levels of integrin subunits alpha4, alpha L, alpha M, and beta2 were significantly higher in NKTCL than non neoplastic controls. Integrin subunits alpha2 and alpha M were expressed at a significantly higher level in NKTCL with angioinvasion than those without. Expression level of alpha M was significantly higher in 7 cases of NKTCL with LMP 1 expression than 7 without. Immunohistochemistry showed expression of these molecules in NKTCL cells. These findings suggest that EBV infection might be involved in the pathogenesis of angioinvasion of NKTCL through up-regulation of alpha M by LMP-1. PMID- 15756433 TI - Thymidine phosphorylase expression and efficacy of adjuvant doxifluridine in advanced colorectal cancer patients. AB - To clarify the correlation between the expression level of thymidine phosphorylase (TP) and efficacy of doxifluridine (5'-DFUR) and 5-fluorouracil (5 FU), samples from 177 colorectal cancer patients who underwent curative resection were evaluated by immunohistochemical staining using a newly developed monoclonal antibody 1C6-203. Patients were randomly given either oral 5'-DFUR or 5-FU as postoperative adjuvant chemotherapy. In Dukes' C staged colon cancer patients treated with 5'-DFUR, better survival was observed in the high TP patients than the low TP patients (P=0.025 by the log-rank test). The observed 5-year survival rates were 91.2 and 74.8%, respectively. No correlation between TP expression and patient prognosis was detected in the 5-FU group. In Dukes' C stage colon patients with high TP expression, the 5'-DFUR group had slightly better survival than the 5-FU group. These findings suggest that TP may be a chemosensitive marker for 5'-DFUR as postoperative adjuvant chemotherapy for advanced colon cancer patients. PMID- 15756435 TI - Detection of Her2/neu, c-MYC and ZNF217 gene amplification during breast cancer progression using fluorescence in situ hybridization. AB - The deletion of tumor suppressor genes and amplification of activating oncogenes appear to be critical events in tumorigenesis. We carried out fluorescence in situ hybridization (FISH) analysis of the breast cancer cell line MCF7 and clinically obtained cancer tissue sections on the basis of which we suggest a breast cancer development model. We selected 28 genes for FISH probes from breast cancer patients. Of the 28 genes studied, 14 were shown to be consistently amplified in the breast cancer cell line MCF7. We selected three genes from clinical tumor samples on the basis of results from MCF7 analysis. The amplification of Her2/neu or ZNF217 is closely associated with the stages of breast cancer. The frequency of amplification of Her2/neu increased notably in patients at stages later than IIB based on TNM staging, whereas the amplification of ZNF217 correlated with T2N1M0 at stage IIB and later stages. c-MYC amplification was not related to the stage. Her2/neu, ZNF217 and c-MYC were found to have a significantly increased copy number in breast cancer cells. In breast cancer progression, c-MYC amplification is an early event, while ZNF217 and Her2/neu amplification may play a role in the later stage of tumor development. PMID- 15756434 TI - Venous thromboembolism in colorectal cancer patients with central venous catheters for 5-FU infusion-based pharmacokinetic modulating chemotherapy. AB - Colorectal cancer patients with central venous catheters (CVC) for pharmacokinetic modulating chemotherapy (PMC) have a substantial risk of venous thromboembolism (VTE). PMC, designed as a hybrid of lower metronomic and higher shorter plasma 5-FU concentrations, has been clinically successful. To determine the effectiveness and safety of D-dimer tests and multidetector-row CT (MDCT) for diagnosis in cancer patients with suspected VTE, we carried out a clinical outcome study on PMC outpatients. Patients received a D-dimer test before and after commencing the PMC regimen. MDCT was performed additionally if the D-dimer test appeared positive or showed signs of VTE. When CT results were positive for thromboembolism, anticoagulation was started. The overall prevalence of VTE in PMC patients was 2.0% (7 of 350 patients). In this study, 34 out of 102 colorectal cancer patients gave a positive D-dimer test (33.3%). CT identified venous thrombi in 2 of the 102 patients (2.0%), mural thrombosis on catheterized veins in another 3 patients (2.9%), and endothelial hyperplasia on catheterized veins in 8 patients (7.8%). The catheters of these patients did not show any significant abnormalities. Patients with negative D-dimer tests showed no signs or symptoms of VTE. In colorectal cancer patients receiving continuous 5-FU infusion via CVC, a D-dimer test can be safely used as the primary diagnostic test for ruling out VTE. We suggest 7.0 microg/ml as the D-dimer cut-off value. Thromboprophylaxis should be considered in the patients showing values >7.0 microg/ml. PMID- 15756436 TI - Expression of c-kit (CD117) in neuroendocrine tumours--a target for therapy? AB - C-kit is a tyrosine kinase receptor which is expressed in a wide variety of tumour cells such as gastrointestinal stromal tumours (GISTs), germ cell tumours, malignant trans-formation of mast cells, breast adenocarcinomas, malignant melanomas and small cell lung cancers. Imatinib mesylate is a tyrosine kinase inhibitor initially developed against the bcr-abl fusion protein of CML, but also shows therapeutic inhibitory activity against c-Kit expressed in GISTs. Treatment of patients with neuroendocrine tumours (NETs) at present is limited. Our aim was to test NETs for c-Kit expression and hence identify patients for the consideration of therapy with imatinib mesylate. NET patient specimens (n=85) were assessed for expression of c-KIT proto-oncogene (CD117) by immunohistochemistry using two antibodies, a polyclonal antibody and a monoclonal. Of the samples 24% stained positive with the polyclonal antibody and 64% with the monoclonal antibody. This study highlights problems related to screening using c-kit antibodies for immunocytochemistry. It is possible that the polyclonal antibody is less specific. Studies need to be performed to determine if c-kit expression by NETs can be translated into therapeutic benefit by agents such as imatinib mesylate. PMID- 15756437 TI - Atypical lipomatous tumor with rare structural rearrangements involving chromosomes 8 and 12. AB - Atypical lipomatous tumor (ALT), an intermediate malignant neoplasm of soft tissues, is characterized by the presence of supernumerary ring and giant marker chromosomes. These supernumerary chromosomes consistently contain amplified 12q material in association with amplified segments from a variety of other chromosomes. However, a few cases of ALT with other types of chromosomal rearrangements have been reported earlier. We report on new types of structural aberrations in a case of ALT. In a pseudodiploid karyotype, there were two aberrant chromosomes, both consisting of alternating chromosome 8 and 12 sequences as shown by multicolor fluorescence in situ hybridization (FISH). The complex rearrangement was not only the result of multiple breaks and reunions of these chromosomes, but was also associated with a gain of chromosome 12 sequences. FISH analyses revealed that the number of MDM2 signals was slightly elevated (median, 5). There were three intact copies of HMGA2 and one additional copy of the 5' part of the gene. These findings are consistent with previous reports that the ALT phenotype may be associated with a low or moderate level of gene amplification, whereas truncation of HMGA2 has been observed in both ALTs and benign lipomas. The aberrations in the present case were stable, although rare cells with higher MDM2 copy numbers were detected. Whether ALTs with these types of aberrations have a lower risk of tumor progression than ALTs with the notoriously mitotically unstable ring and giant marker chromosomes remains to be investigated. PMID- 15756438 TI - Antiproliferation effects of ponicidin on human myeloid leukemia cells in vitro. AB - Ponicidin, an extract from the Chinese herb Rabdosia rubescens, is currently one of the most important traditional Chinese herbal medicines. Ponicidin has been reported to have anti-tumor effects on a large variety of malignant diseases. In this study, we investigated the anti-proliferation effects of ponicidin on human myeloid K562 and HL-60 cells. Cell viability was measured by MTT assay; cell apoptosis was assessed by flow cytometry, DNA fragmentation analysis and Hoechst 33258 staining. Caspase-3 and poly(ADP-ribose) polymerase (PARP) activation and Bax and Bcl-2 expression were detected by Western blot analysis. The results revealed that ponicidin could significantly inhibit the growth of K562 and HL-60 cells by induction of apoptosis. The suppression was both time- and dose dependent. Cell apoptosis was observed clearly after the cells were treated with ponicidin for 48-72 h. Western blotting showed cleavage of the caspase-3 zymogen protein (32 kDa) with the appearance of its 17 kDa subunit, together with a cleaved 89-kDa fragment of 116 kDa PARP when apoptosis occurred. Bcl-2 expression was down-regulated while Bax expression up-regulated concurrently when the cells were treated with ponicidin for 24-48 h. Therefore, we conclude that ponicidin has significant anti-proliferation effects by induction of apoptosis on myeloid leukemia cells in vitro, down-regulation of Bcl-2, and up-regulation of Bax, and that activation of caspase-3 and PARP may be an important apoptosis-inducing mechanism. The results suggest that ponicidin may serve as a potential therapeutic agent for leukemia. PMID- 15756439 TI - Inhibition of cell cycle and induction of apoptosis by sulforaphane in cell lines carrying various inherited BRCA1 mutations. AB - The important aspect of sulforaphane (SFN) chemopreventive activity is its ability to induce cell growth inhibition and apoptosis. In this study, the effect of SFN on lymphoblastoid cells derived from people carrying four different germ line mutations in BRCA1 gene was tested and compared to the effect of SFN on wild type cells. The mutations studied were C61G, 3819del5, 4153delA and 5382INSC. Changes in cell viability and density after SFN treatment were evaluated, as well as cell cycle progression, changes in mitochondrial membrane potential, and phosphatidylserine externalization. SFN was shown to reduce cell viability and density in all cell lines tested. Cell cycle block in S-phase and the occurence of simultaneous apoptosis were observed. The concentration of SFN needed to elicit a comparable effect on each cell line was varied. We found that the effect of SFN on cells carrying different inherited mutations depended on mutation type. PMID- 15756440 TI - Long-term administration of low-dose cisplatin plus 5-fluorouracil prolongs the postoperative survival of patients with esophageal cancer. AB - To evaluate the efficacy of long-term postoperative adjuvant chemotherapy with low-dose cisplatin (CDDP) plus 5-fluorouracil (5-FU) (CDDP/5-FU), we retrospectively examined 167 patients with squamous cell carcinoma of the esophagus who received the treatment after curative surgery (R0 resection). We classified the patients into the following three groups according to their postoperative therapies and analyzed their outcomes: a) low-dose CDDP (10 mg body(-1) day(-1) x 5 days) plus 5-FU (250-500 mg body(-1) day(-1) x 5 days) repeated every 6 months for 3 years, with an oral fluoropyrimidine (5-FU 150-200 mg body(-1) day(-1) or UFT 300-400 mg body(-1) day(-1)) administered between each treatment cycle (low-dose CDDP/5-FU group, 98 patients); b) high-dose CDDP (80 mg body(-1) day(-1) x 1 day) plus 5-FU (750-1,000 mg body(-1) day(-1) x 5 days) administered once only, followed by treatment with an oral fluoropyrimidine (5-FU 150-200 mg body(-1) day(-1) or UFT 300-400 mg body(-1) day(-1)) for 3 years (high dose CDDP/5-FU group, 17 patients); or c) surgery alone (surgery alone group, 52 patients). The 3-year survival rates were 83.7% in the low-dose CDDP/5-FU group, 61.4% in the high-dose CDDP/5-FU group, and 62.2% in the surgery alone group; the difference between the low-dose CDDP/5-FU group and surgery alone group was significant (log-rank, p<0.05). A significantly better outcome in the low-dose CDDP/5-FU group than in the surgery alone group was associated with pStage III disease (p<0.001), pN1 lymph node metastasis (p<0.001), and lymphatic invasion (p<0.01). We conclude that long-term postoperative treatment with low-dose CDDP/5 FU is therapeutically beneficial and prolongs survival in patients with esophageal cancer who have regional lymph node metastasis or lymphatic invasion. PMID- 15756441 TI - Aberrant expression of HOX genes in human invasive breast carcinoma. AB - HOX genes are known not only as master genes that control the morphogenesis, but also as regulator genes that maintain tissue or organ specificity in the adult body. We hypothesized that dysregulated expression of HOX genes was associated with tumor development and malignant progression such as invasion and metastasis. In this study, we analyzed the expression patterns of 39 HOX genes in human invasive ductal breast cancer tissues and normal tissues by the real-time RT-PCR method. We found 11 HOX genes (HOXA1, A2, A3, A5, A9, C11, D3, D4, D8, D9 and D10) expression levels of which were significantly different between cancerous and normal tissues. All 10 genes except HOXC11 were expressed at lower levels in cancerous tissues than normal tissues. Comparing expression levels of each HOX gene among the different types of cancer tissues, the expression level of HOXB7 was lower in lymph node metastasis-positive cancer tissues than negative cancer tissues; those of HOXD12 and D13 were higher in progesterone receptor-positive cancer tissues than negative cancer tissues; and the expression level of HOXC5 was lower in cancerous tissues with mutated-type p53 than in normal and cancerous tissues with wild-type p53. These results suggest that the aberrant expression of HOX genes is related to the development of breast cancer and malignant behavior of cancer cells. PMID- 15756442 TI - Irinotecan combined or alternated with bolus 5-fluorouracil and folinic acid versus the Mayo Clinic regimen in the first-line therapy of advanced colorectal cancer. AB - The objective of this study was to assess the efficacy and safety of two regimens of irinotecan, combined or alternated with bolus 5-fluorouracil (5-FU) and folinic acid (FA), and the Mayo Clinic regimen as first-line therapy for colorectal cancer (CRC). A total of 152 patients with advanced CRC were randomised, and 149 patients were treated intravenously by irinotecan 125 mg/m2, FA 20 mg/m2 followed by 5-FU 500 mg/m2 bolus, weekly for 4 weeks (arm A, Saltz regimen; n=46), or irinotecan 350 mg/m2 alternating with FA 20 mg/m2/day followed by 5-FU bolus 425 mg/m2/day for 5 days (arm B; n=53), or FA 20 mg/m2/ day followed by 5-FU bolus 425 mg/m2/day over 5 days every 4 weeks (arm C, Mayo Clinic regimen; n=50). Patients were analyzed for tumor response, time to progression, overall survival, safety and quality of life. The overall response rate for evaluable patients in arm A was 33% [95% confidence interval (CI), 17 49%], in arm B was 32% (95% CI, 16-49%) and in arm C was 26% (95% CI, 12-40%). Median times to progression were 7.9, 7.0 and 6.9 months and median survival times were 22.2, 17.0 and 18.2 months for arms A, B and C, respectively, in the intention-to-treat population. The main grade 3-4 adverse events were neutropenia (7%, 39% and 12%) and diarrhea (6%, 21% and 18%). In conclusion, both regimens containing irinotecan were active and well tolerated in patients with advanced CRC. PMID- 15756443 TI - Pro-neoangiogenic cytokines (VEGF and bFGF) and anemia in solid tumor patients. AB - Using a commercial ELISA assay, we evaluated circulating VEGF and bFGF levels in 203 consecutive patients with solid tumors, and sought a correlation between them and with the grade of anemia. Serum VEGF values were within the normal range in 128 patients (63.05%), with a mean value of 675.04 pg/ml (median, 571.00; range, 0-2796.54). The analysis of VEGF values per tumor group did not provide any statistically significant difference. Regarding bFGF, 143 patients (70.44%) had measurable, and thus abnormal, bFGF values. Overall, mean bFGF serum value was 57.14 pg/ml (median, 8.30; range, 0-4334.71), with the highest bFGF levels found in breast carcinoma patients. As expected, a large number of our patients was fairly anemic, mean hemoglobin level being 11.47 g/dl (median, 11.30; range, 7.1 19.20), the lowest titers being observed in prostate carcinoma patients. No statistically significant correlation was found between serum VEGF and hemoglobin values (r=0.004) but a significant negative correlation was seen between serum bFGF and hemoglobin (r=-0.22, p<0.05). Considering the different tumor groups, a statistically significant negative correlation between bFGF and hemoglobin becomes even more apparent in the subgroup of renal carcinoma patients (r=-0.55, p<0.05). In conclusion, our results demonstrate that there is a statistically significant correlation between systemic hypoxia (evaluated in terms of hemoglobin levels) and circulating bFGF values, but not VEGF; this correlation may lead to therapeutic interventions. PMID- 15756444 TI - Predictive value of COX-2 for the effect of chemoradiotherapy on esophageal squamous cell carcinoma. AB - Cyclooxygenase-2 (COX-2) displays anti-apoptotic functions related to angiogenesis or blocking of bcl-2 functions. COX-2 overexpression has been found in various human malignancies, including esophageal squamous cell carcinoma (ESCC). The present study examined correlations between expression of COX-2 mRNA and ESCC responses to chemoradiotherapy (CRT). Expression of COX-2 mRNA obtained from 29 biopsy specimens before CRT was quantified using reverse transcriptase polymerase chain reaction (RT-PCR) using Sybr Green I on the Roche LightCycler system. CRT comprised 5-fluorouracil plus cisplatin and 40 Gy of radiation. Mean COX-2 mRNA score was significantly higher in tumors (1222) than in normal epithelium (50; p=0.05). Expression of COX-2 mRNA was high in 14 patients, low in 8 and absent in 7. The effective response to CRT was achieved in 18 patients. Mean COX-2 mRNA score was significantly higher in ineffective cases (2910) than in effective cases (190; p<0.05). CRT was more effective in patients with low COX 2 mRNA expression than in those with high expression. COX-2 mRNA expression in biopsy specimens was closely related to CRT effectiveness. Examination of COX-2 mRNA expression is useful for predicting the effect of CRT in patients with ESCC. PMID- 15756445 TI - Infrequent mutation of the human envoplakin gene is closely linked to the tylosis oesophageal cancer locus in sporadic oesophageal squamous cell carcinomas. AB - Envoplakin (EVPL) is a member of the desmosomal plaque proteins attached to desmosomal cadherin and keratin filaments. The EVPL gene has been mapped to the tylosis oesophageal cancer (TOC) locus on chromosome 17q25, where it has been demonstrated to be frequently deleted in both familial and sporadic forms of oesophageal squamous cell carcinoma (OSC). In this study, we examined EVPL gene mutations in 10 OSC cell lines and 20 sporadic OSCs using reverse transcription polymerase chain reaction single-strand conformational analysis (RT-PCR SSCP) followed by direct sequencing. We observed one somatic mutation (GCG to ACG at codon 1104, Ala to Thr: 1/20, 5%) in the central rod domain and 5 intragenic polymorphic sites, where frequent loss of heterozygosity (LOH) (63%) was detected. No mutations were detected in the OSC cell lines. The rate of EVPL gene mutation was quite low in contrast to the frequency of LOH on the TOC locus in sporadic OSCs, and the high incidence of oesophageal cancer development in tylosis families. Our results suggest that EVPL might not be the target gene responsible for OSC, despite its strong candidacy in terms of character and localization. PMID- 15756446 TI - Establishment and gene analysis of a cisplatin-resistant cell line, Sa-3R, derived from oral squamous cell carcinoma. AB - Cisplatin (CDDP) is widely used for chemotherapy of many malignancies, especially of oral squamous cell carcinoma (SCC). However, because the mechanism of resistance to CDDP is unclear, we established a CDDP-resistant cell line, Sa-3R, from a CDDP-sensitive cell line, Sa-3, which was derived from moderately differentiated SCC of the lower gingiva. The 3-(3,4-dimethyl-thiazol-2-yl) 2,5 diphenyltetrazolium bromide assay indicated that Sa-3R has 7.5-fold greater resistance to CDDP than Sa-3. Comparing gene expression levels in the cell lines using an in-house cDNA microarray, which represented 2,201 oral disease origin genes, many differentially expressed genes were identified. The ATP-binding cassette transporter genes (MDR-1, MRP-1, and MRP-2), and FANCONI, GRP58, FLJ12089, and SPINT-2 were up-regulated, whereas FOSL1, MRPS27, and PGK-1 were down-regulated. These results were confirmed by semiquantitative reverse transcriptase-polymerase chain reaction. The Sa-3/Sa-3R cell lines could be useful to identify the candidates responsible for the mechanism of CDDP resistance and the up- or down-regulated genes identified by the gene expression profiles in the Sa-3R cell line may be, in part, associated with the mechanism. PMID- 15756447 TI - E1AF expression is closely correlated with malignant phenotype of tongue squamous cell carcinoma through activation of MT1-MMP gene promoters. AB - E1AF is an ets-oncogene family transcription factor that has been shown to up regulate multiple MMPs whereas MMP-2, a potent extracellular matrix degrading enzyme, is not up-regulated. We investigated the activation mechanism of MMP-2 in oral squamous cell carcinoma. Chloramphenicol acetyltransferase (CAT) assay was utilized to investigate whether E1AF is able to up-regulate membrane type-1 matrix metalloproteinase (MT1-MMP), which is known to activate MMP-2. Expression of the CAT reporter gene under the control of the MT1-MMP promoter was increased approximately 40-fold by co-transfection with the E1AF expression vector. Real time RT-PCR study was carried out in 25 patients with tongue squamous cell carcinoma, and the mRNA expression level of E1AF and MT1-MMP was synergistically increased. These results indicate that E1AF positively regulates transcription from MT1-MMP genes, which plays an important role in invasion and metastasis of squamous cell carcinoma of the tongue by converting pro-MMP-2 into active-MMP-2. PMID- 15756448 TI - Regulation of tumor invasion by HOXB13 gene overexpressed in human endometrial cancer. AB - During the last two decades, a group of homeobox-containing genes, the HOX gene family, has been studied both in the context of embryonic development and neoplasia. In particular, there is accumulating evidence of the involvement of HOX abnormalities in a variety of malignancies, including breast cancer. However, little is known about the association of HOX genes with endometrial cancer, which is the most common malignancy of the female genital tract and is thought to be dependent on estrogen, like breast cancer. In this study, we detected overexpression of the HOXB13 gene in endometrial cancer cells and tissues from patients by quantitative real-time RT-PCR. To investigate whether overexpression of HOXB13 is involved in invasion or metastasis of endometrial cancer, we transfected antisense HOXB13/pcDNA3.1+ plasmid vector into endometrial cancer AN3CA cells by electroporation and performed in vitro chemoinvasion assay. We revealed that the invasive ability of antisense-transfectants showed a 90% reduction compared with parental cells and control transfectants (p<0.01). In addition, administration of 17beta-estradiol induced time- and dose-dependent responses of the HOXB13 expression in endometrial cancer AN3CA cells. These results suggest that overexpression of HOXB13 in endometrial cancer may be associated with the invasive ability of cancer cells with regulation by estrogen. PMID- 15756449 TI - The prognostic value of overexpression of Skp2 mRNA in non-small cell lung cancer. AB - Skp2 putatively plays a critical role in regulating cell cycle progression. A high level of Skp2 expression is observed in a variety of cancer. No previous report describes the prognostic value of Skp2 mRNA in non-small cell lung cancer (NSCLC). Therefore, the purpose of this study was to examine overexpression of Skp2 mRNA in NSCLC and identify the association of Skp2 expression level with patient survival. We investigated Skp2 expression in 79 patients with NSCLC by means of reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The Skp2 expression was significantly associated with tumor status, lymph node metastasis, stage, vascular invasion, histology and Skp2 protein. With regard to prognosis, the overall and stage I survival rates for patients in the high Skp2 mRNA group were significantly poorer when compared with the low Skp2 mRNA group. Furthermore, expression of Skp2 mRNA was an independent predictor of prognosis by multivariate analysis. Skp2 expression might play an important role in the development and progression in NSCLC and Skp2 gene expression can serve as a useful prognostic marker in NSCLC patients. PMID- 15756450 TI - High lymphatic vessel density correlates with overexpression of VEGF-C in gastric cancer. AB - Lymph node metastasis is one of the most important prognostic factors in malignant tumors. In this study, we investigated vascular endothelial growth factor (VEGF)-C expression in human gastric cancer using immunohistochemical techniques and determined the number of microvessels in peritumoral tissue. VEGF C expression was positive in 22 of 79 cases (27.8%), and correlated with the presence of lymphatic invasion and lymph node metastasis. We confirmed by reverse transcription-polymerase chain reaction (RT-PCR) that VEGF-C mRNA expression is observed more commonly in cancer tissues than normal tissues. For 59 gastric tumors, we examined lymphatic vessel density (LVD) using the specific lymphatic vessel endothelial hyaluronan receptor (LYVE) -1 antibody. VEGF-C expression was observed in 10 of 25 cases (40%) that exhibited a high LVD. Furthermore, high LVD exhibited a significant correlation with VEGF-C expression. Our findings suggest that VEGF-C plays a pivotal role for lymphangiogenesis and tumor growth in gastric cancer. PMID- 15756451 TI - Loss of heterozygosity in ductal lavage for breast tumor and the contralateral breast. AB - Detection of breast cancer at an early stage is the key to successful treatment and outcome. In tumors confined to the breast at diagnosis, stage is the most important prognostic factor. If detected early, breast cancer is eminently curable. We measured loss of heterozygosity (LOH) using 9 microsatellite markers in epithelial cells obtained by ductal lavage (DL) from 59 women before breast surgery (excisional biopsies, duct resections, and mastectomies). In addition, we analyzed LOH obtained by DL in the contralateral breast of breast cancer patients. As a result, histological examination revealed 46 malignancies and 13 benign lesions. Sensitivity, specificity, and diagnostic accuracy were 100.0, 15.4, and 82.3% for LOH. LOH frequency in DL of breast cancer was significantly greater than that in DL of benign tumors. In terms of the contralateral breast of breast cancer patients, DL showed that one subject (3.4%) was malignant, and LOH frequency was similar to breast carcinoma. In conclusion, breast tissues that appear non-neoplastic pathologically often harbor genetic changes that can be important in understanding the breast environment within which the cancer develops. PMID- 15756452 TI - Role of extracellular matrix in regulation of staurosporine-induced apoptosis in breast cancer cells. AB - Autocrine and paracrine mechanisms modulate the synthesis and secretion of extracellular matrix (ECM); moreover, each component of the ECM is capable of modulating the synthesis and release of other ECM molecules. Therefore, the synthesis of ECM glycoprotein fibronectin and laminin was studied in the human breast cancer cell lines MCF7 and MDA MB 23, plated on different ECM. Our results showed that the cells plated on a fibronectin substrate increased laminin synthesis: this event correlated with an increase in alpha2 and alpha3 integrin subunits. Staurosporine-induced apoptosis was then analyzed in the cell lines plated on different ECM. Staurosporine treatment determined the apoptosis of 35 and 33% respectively of MDA MB 231 and MCF7; these values increased to 60 and 64% in cells plated on laminin, to 48 and 63% in cells plated on fibronectin and to 64 and 69% in cells plated on matrigel. Moreover, staurosporine treatment decreased bcl-2 expression in the cells plated on fibronectin and laminin. Yet, staurosporine treatment determined PARP cleavage and PARP partial disappearance when the cells were plated on matrigel. Finally, a partial loss of function mutant Ras protein that activated only Raf pathway, was expressed in MCF7, in order to identify whether the increase of apoptosis induced by extracellular matrix involved the Raf/MAP kinase pathway. The increase of apoptosis of the cells plated on matrigel suggested that the activation of the Raf pathway is probably involved in the decrease of survival on matrigel. These data demonstrate that the modification of ECM modulates the apoptotic process of breast cancer cells and suggest that it is worthwhile to dissect the role of ECM in the control of apoptotic process. PMID- 15756453 TI - Immunotherapy of renal cell carcinoma with granulocyte macrophage colony stimulating factor and very low dose interleukin-2. AB - We have performed a translational phase II trial testing an original immunotherapy schedule based on the monthly subcutaneous (s.c.) administration of hrGM-CSF (days 1 through 5) and very low dose hrIL-2 (days 6 through 15) in 19 patients with metastatic renal cell carcinoma. Bone pain, first dose reaction to GM-CSF, asthenia and fever were the most common side effects. A partial response, and a disease stabilization were respectively observed in 4 and 11 cases, with a rate of objective response and a disease control rate respectively of 21% and 79%. We recorded a time to progression of 9 months and a 2- and 3-year survival respectively of 42% (8/19 patients) and 26% (5/19 patients). Our results suggest that this GM-CSF/hrIL-2 combination is active and well tolerated in patients with renal cell carcinoma and deserves to be investigated in larger comparative trials. PMID- 15756454 TI - Selection of tumor-targeting agents on freshly excised human breast tumors using a phage display library. AB - The selective delivery of therapeutic agents to tumor site without harming rest of the body is a major challenge in clinical oncology today. Phage display approach has been used for searching ligands for cell-surface macromolecules on cancer cells so that they can be employed as drug targeting agents. Although basic protocols for biopanning cells are available, they are not as such suitable for screening highly complex and diverse target as whole tumor. Present study is an attempt to select peptide ligands specific to whole tumors. The cells from freshly collected human breast tumors were biopanned with phage displayed disulfide-constrained random heptapeptide library, following subtraction on normal human breast cells. Comparative analysis of amino acid frequencies in tumor-selected peptides and in random peptides from unselected library showed that selection was not random. The binding assessment of tumor-selected clones, using highly sensitive chemiluminescence ELISA, demonstrated that 47-75% of selected clones, depending on a tumor, bound to tumor cells they were panned on. Furthermore, several clones bound exclusively or preferentially to tumor cells in comparison to normal breast cells. It was interesting to note that insert sequences of tumor-binding clones from different tumors shared significant motifs. It shows the possibility of identifying ligands that may bind to tumor specific targets common in certain tumors. The results of this investigation on seven human breast tumors demonstrated that, using procedures developed in the present study, whole tumors can be panned successfully with phage displayed library and tumor-binding ligands can be identified rapidly in high throughput manner. This is an important enabling step in identifying lead molecules for developing novel, specific, and effective agents that can be used for the diagnosis and treatment of cancer. PMID- 15756455 TI - Human antibody SC-1 reduces disseminated tumor cells in nude mice with human gastric cancer. AB - Advanced gastric cancer is a systemic disease that requires adjuvant therapy targeted at eliminating disseminated tumor cells (DTCs). We investigated whether the apoptosis-inducing human monoclonal IgM antibody SC-1 was able to reduce the number of disseminated gastric cancer cells in blood and bone marrow. Human gastric tumor specimens with positive expression of the SC-1 receptor were transplanted in nude mice with metastasizing gastric cancer. After tumor growth (4-6 weeks) animals were randomly allocated to intraperitoneal 100 microg SC-1 (n=23) or 100 microg human IgM (n=23). One week later, animals were sacrificed and blood and bone marrow specimens were obtained. A nested RT-PCR for cytokeratin 20 (CK-20) from blood and bone marrow of mice was performed for detection of disseminated tumor cells. Animals receiving SC-1 had significantly fewer DTCs than did control animals (p=0.0011). None of the SC-1 mice had DTCs simultaneously in both blood and bone marrow versus four of the control animals (p=0.0363). The reduction of DTCs in SC-1 animals was due to reduction in bone marrow (p=0.032 compared to controls), but not in blood (p=0.1158). Treatment with SC-1 significantly reduced the number of DTCs in bone marrow in this animal model. PMID- 15756456 TI - Comparative genomics on Wnt16 orthologs. AB - The WNT16 gene at human chromosome 7q31.31 encodes two isoforms due to alternative splicing of an alternative promoter type. Here, we identified and characterized the rat Wnt16 and zebrafish wnt16 genes using bioinformatics. The rat Wnt16 and zebrafish wnt16 genes, consisting of four exons, were located within the AC117014.4 and CR925699.1 genome sequences, respectively. Exons 1b, 2, 3 and 4 of the human WNT16 gene were conserved in the rodent Wnt16 and zebrafish wnt16 genes; however, exon 1a of the human WNT16 gene was not conserved. Human WNT16 isoform 1 (NM_057168.1), consisting of exons 1b, 2, 3 and 4, was identified as the evolutionarily conserved representative isoform. Because zebrafish NM_207096.1 was an aberrant wnt16 cDNA with a retained 3'-part intron 1, complete CDS of representative zebrafish wnt16 was determined by deleting nucleotide position 183-239 from NM_207096.1 aberrant cDNA. Rat Wnt16 (364 aa) with an N terminal signal peptide, 24 Cys residues and 3 Asn-linked glycosylation sites showed 97.3, 90.4 and 65.9% total-amino-acid identity with mouse Wnt16, human WNT16, and zebrafish wnt16, respectively. Phylogenetic analyses on WNT family members revealed that WNT16 was most closely related to WNT7A and WNT7B paralogs. Promoters of human WNT16, rat Wnt16 and mouse Wnt16 genes were well conserved. Double CCAAT motifs were conserved among mammalian Wnt16 promoters. This is the first report on the rat Wnt16 and zebrafish wnt16 genes, as well as the double CCAAT motifs within the core promoter regions of mammalian Wnt16 orthologs. PMID- 15756458 TI - (13)C-(13)C NOESY: a constructive use of (13)C-(13)C spin-diffusion. AB - (13)C-(13)C NOESY experiments were performed under long mixing time conditions on reduced human superoxide dismutase (32 kDa, (15)N, (13C) and 70% (2)H labeled). (13)C-(13)C couplings were successfully eliminated through post-processing of in phase-anti-phase (IPAP) data. It appears that at mixing time tau(m) of 3.0 s the spin diffusion mechanism allows the detection of 96% of the two-bond correlations involving C' and C(beta). The interpretation was confirmed by simulations. This approach broadens the range of applicability of (13)C-(13)C NOESY spectroscopy. PMID- 15756457 TI - Comparative genomics on Wnt7a orthologs. AB - WNT and Hedgehog signaling pathways network together during carcinogenesis and embryogenesis. WNT7A mRNA is expressed in human gastric and pancreatic cancer cells. WNT7A mRNA expression is relatively high in the temporal, occipital and parietal lobes, paracentral gyrus of the cerebral cortex, caudate nucleus, hippocampus, medulla oblongata and putamen within the adult human brain. Here, we identified and characterized the rat Wnt7a gene using bioinformatics. The rat Wnt7a gene, consisting of four exons, was located within the AC106100.7 genome sequence. Rat Wnt7a (349 aa) with 24 Cys residues and 3 Asn-linked glycosylation sites showed 99.7, 99.1 and 93.7% total-amino- acid identity with mouse Wnt7a, human WNT7A and chicken wnt7a, respectively. The N-terminal signal peptide was not identified within the Wnt7a orthologs. Exonic regions, except the 3'-UTR, were conserved between the rat Wnt7a and human WNT7A genes; however, the 5' flanking region was significantly divergent between the rat Wnt7a and human WNT7A genes. Although the COMP1- and ELK1-binding sites within exon 1 were conserved among mammalian Wnt7a promoters, transcriptional regulation of the rodent Wnt7a orthologs was predicted to be distinct from that of human WNT7A based on the divergence within the 5'-flanking promoter region. Therefore, biological functions of rodent Wnt7a are not always consistent with those of human WNT7A due to promoter evolution. This is the first report on the rat Wnt7a gene and comparative genomics for Wnt7a orthologs. PMID- 15756459 TI - A straight-forward method of optimising protein solubility for NMR. AB - Maximising solubility is a key step in applying solution-state NMR to proteins. The 'microbatch' crystallisation screening method can be adapted to screen for protein solubility. In this approach, drops of test solutions are placed under paraffin oil in 96-well screening plates. This requires very small amounts of protein, is easy to set up and is readily automatable. PMID- 15756460 TI - Resolution-optimized NMR measurement of (1)D(CH), (1)D(CC) and (2)D(CH) residual dipolar couplings in nucleic acid bases. AB - New methods are described for accurate measurement of multiple residual dipolar couplings in nucleic acid bases. The methods use TROSY-type pulse sequences for optimizing resolution and sensitivity, and rely on the E.COSY principle to measure the relatively small two-bond (2)D(CH) couplings at high precision. Measurements are demonstrated for a 24-nt stem-loop RNA sequence, uniformly enriched in (13)C, and aligned in Pf1. The recently described pseudo-3D method is used to provide homonuclear (1)H-(1)H decoupling, which minimizes cross correlation effects and optimizes resolution. Up to seven (1)H-(13)C and (13)C (13)C couplings are measured for pyrimidines (U and C), including (1)D(C5H5), (1)D(C6H6), (2)D(C5H6), (2)D(C6H5), (1)D(C5C4), (1)D(C5C6), and (2)D(C4H5). For adenine, four base couplings ((1)D(C2H2), (1)D(C8H8), (1)D(C4C5), and (1)D(C5C6)) are readily measured whereas for guanine only three couplings are accessible at high relative accuracy ((1)D(C8H8), (1)D(C4C5), and (1)D(C5C6)). Only three dipolar couplings are linearly independent in planar structures such as nucleic acid bases, permitting cross validation of the data and evaluation of their accuracies. For the vast majority of dipolar couplings, the error is found to be less than +/-3% of their possible range, indicating that the measurement accuracy is not limiting when using these couplings as restraints in structure calculations. Reported isotropic values of the one- and two-bond J couplings cluster very tightly for each type of nucleotide. PMID- 15756461 TI - Addressing the overlap problem in the quantitative analysis of two dimensional NMR spectra: application to (15)N relaxation measurements. AB - A quantitative analysis of 2D (1)H-(15)N spectra is often complicated by resonance overlap. Here a simple method is presented for resolving overlapped correlations by recording 2D projection planes from HNCO data sets. Applications are presented involving the measurement of (15)N T(1rho) relaxation rates in a high molecular weight protein, malate synthase G, and in a system that exchanges between folded and unfolded states, the drkN SH3 domain. By supplementing relaxation data recorded in the conventional way as a series of 2D (1)H-(15)N data sets with a series of a pair of projection planes the number of dynamics probes is increased significantly for both systems studied. PMID- 15756462 TI - (1)H, (13)C and (15)N backbone assignment of a 32 kDa hypothetical protein from Arabidopsis thaliana, At3g16450.1. PMID- 15756463 TI - (1)H, (13)C and (15)N chemical shift assignments of an enolase-phosphatase, E1, from Klebsiella oxytoca. PMID- 15756464 TI - Backbone assignment of the dimerization and DNA-binding domain of the oncogenic transcription factor v-Myc in complex with its authentic binding partner Max. PMID- 15756465 TI - (1)H, (13)C and (15)N resonance assignment of the reduced form of methionine sulfoxide reductase A from Escherichia coli. PMID- 15756466 TI - (1)H, (13)C and (15)N resonance assignments of coactosin, a cytoskeletal regulatory protein. PMID- 15756467 TI - Resonance assignments for the endosomal adaptor protein p14. PMID- 15756468 TI - Assignment of the (1)H, (13)C and (15)N resonances of the class II E2 conjugating enzyme, Ubc1. PMID- 15756469 TI - (1)H, (15)N and (13)C chemical shift assignments of RNA repeats binding protein - CUGBP1ab. PMID- 15756470 TI - (1)H, (13)C and (15)N resonance assignments of Ca(2+)-free DdCAD-1: a Ca(2+) dependent cell-cell adhesion molecule. PMID- 15756471 TI - (1)H, (13)C, and (15)N chemical shift assignment of the C-terminal 15 kDa domain of a novel galactose-binding protein from the earthworm Lumbricus terrestris. PMID- 15756472 TI - Kinetics of phase separation in ramified polymer blends of arbitrary topology. AB - We present here a theoretical study of kinetics of phase separation within a mixture made of two chemically incompatible ramified polymers. For simplicity, we assume that they have the same topology. We are interested in the variation of the relaxation rate, tau(q), versus the wave number q, in the vicinity of the spinodal temperature. The kinetics is governed by local (Rouse) and reptation motions (faster and slower modes). For qRG >> 1 (RG being the gyration radius), kinetics is entirely controlled by local motions where each chain moves inside its own tube, and we show that the corresponding characteristic frequency, tau(q)(-1), scales as tau(q)(-1) approximately kappaGq6, where kappaG is a known topological factor. For qR(G) << 1, however, kinetics is rather dominated by long wavelength (reptation) motions where unlike ramified polymers creep inside a long tube. For this case, we find that tau(q)(-1) approximately delta (0) q2 (chi(c) - chi), where delta (0) is another known topological factor that represents the total mobility of free monomers belonging to connected chains and reticulation points, and chi(c) accounts for the critical value of the segregation parameter. Finally, the derived relaxation rate must be compared to that relative to a linear polymer mixture. PMID- 15756473 TI - Kinetics of microphase separation in interpenetrated polymer networks in solution. AB - We present here a theoretical study of the early kinetics of the microphase separation in crosslinked polymer blends, made of two incompatible polymers A and B, dissolved in a common good solvent. Use is made of an extended blob model used previously for the investigation of the static properties of such a transition. We are interested in the variation of the relaxation rate, tau(q), versus the wave number q, in the vicinity of the spinodal temperature. We first show that kinetics is entirely dominated by local motions, which are of Rouse type. Slow motions are absent, because of the permanent presence of crosslinks. Second, we find that the characteristic frequency, omega (q) = tau(q)(-1), increases with increasing wave number q according to a sixth power law, that is omega (q) approximately q6 phi(-9/4), where phi is the overall monomer volume fraction. Therefore, the swelling of strands due to the excluded-volume forces leads to a renormalization of the characteristic frequency by a multiplicative factor scaling as phi(-9/4). The main conclusion is that the presence of a good solvent necessitates relaxation rates less important than those relative to crosslinked mixtures in the molten state. PMID- 15756474 TI - Specific detection of Pacific oyster (Crassostrea gigas) larvae in plankton samples using nested polymerase chain reaction. AB - Management of sustainable Pacific oyster fisheries would be assisted by an early, rapid, and accurate means of detecting their planktonic larvae. Reported here is an approach, based on polymerase chain reaction (PCR), for the detection of Pacific oyster larvae in plankton samples. Species-specific primers were designed by comparing partial mitochondrial cytochrome oxidase subunit I (COI) sequences from Crassostrea gigas, with other members of the family Ostreidae including those of Crassostrea angulata. Assay specificity was empirically validated through screening DNA samples obtained from several species of oysters. The assay was specific as only C. gigas samples returned PCR-positive results. A nested PCR approach could consistently detect 5 or more D-hinge-stage larvae spiked into a background of about 146 mg of plankton. The assay does not require prior sorting of larvae. We conclude that the assay could be used to screen environmental and ballast water samples, although further specificity testing against local bivalve species is recommended in new locations. PMID- 15756475 TI - Gene expression in tilapia following oral delivery of chitosan-encapsulated plasmid DNA incorporated into fish feeds. AB - DNA delivery into fish is important for transient gene expression, (e.g., DNA vaccination). Previous studies have generally focused on intramuscular injection of DNA vaccines into fish. However, this method is obviously impractical and laborious for injecting large numbers of fishes. This study reports oral delivery of a construct expressing the beta-galactosidase reporter gene into fish by encapsulating the DNA in chitosan and incorporating it into fish feeds. We found that beta-galactosidase expression could be observed in the stomachs, spleens, and gills of fishes fed with flakes containing the chitosan-DNA complex. These results suggest that DNA vaccines and other constructs can be easily and cheaply delivered into fishes orally by use of carriers and incorporation into fish feeds. PMID- 15756476 TI - Variability of ribosomal DNA ITS-2 and its utility in detecting genetic relatedness of pearl oyster. AB - The objective of this study was to detect interspecific and intraspecific genetic variations of the second internal transcribed spacer of ribosomal DNA (ITS-2), and explore the feasibility of using it as a molecular marker phylogenetic analyses and species identification among pearl oysters. ITS-2 sequences of 6 pearl oysters were amplified via polymerase chain reaction. The amplified DNA fragments were about 500 bp, spanning the partial sequences of 5.8S and 28S rRNA genes. The GC contents of all species used in this study were higher than the AT contents. The variations of sequences involved substitutions as well as insertions/deletions and were mainly concentrated in spacer regions. Sequences of about 30-bp in spacer regions showed no variations among 5 Pincatda species. Intraindividual and intraspecific polymorphisms of ITS-2 sequences were detected in some species; the interspecific variability was significantly larger than the variability within species, and the variability at the genus level was higher than that at the species level. Both neighbor-joining and parsimony analyses of ITS-2 sequences revealed the distinguishable species boundary of 6 pearl oysters, and indicated that P. chemnitzi and P. nigra were the closely related species, as were P. maxima and P. margaritifera. The findings revealed that ITS-2 sequences could be an appropriate tool for phylogenetic study of pearl oysters. PMID- 15756477 TI - Cloning and characterization of microsatellite loci in a gorgonian coral, Junceella juncea (Anthozoa; Octocorallia; Ellisellidae) and its application in clonal genotyping. AB - We attempted to isolate microsatellites from a Symbiodinium-free gorgonian octocoral, Junceella juncea, using two methods, partial genomic library screening and enrichment. Among the 3856 clones screened by the partial library method, 10 possibly positive signals were found, and 3 of them could be used to design primers and amplified consistently. In contrast, only one locus isolated by the enrichment method gave reliable amplification and was useful. The results indicate that microsatellites are rare in Junceella juncea, as reported for other cnidarians. Overall, we obtained 4 polymorphic loci to test the feasibility in investigating clonal structure of J. juncea. A total of 40 multilocus genotypes were found among 152 colonies, and the number of genotypes (clones) identified at 7 reefs ranged from 2 to 16. The results of a nonmetric multidimensional scaling analysis indicated the recruitment of J. juncea populations mainly comes from self-retention. These novel microsatellite loci will provide a useful tool to study clonal structure and population genetics for J. juncea in the future. PMID- 15756480 TI - [Aspects of violence in the domestic environment and in institutions]. PMID- 15756478 TI - Recombinant expression and characterization of five-domain Kazal-type serine proteinase inhibitor of black tiger shrimp (Penaeus monodon). AB - Four full-length complementary DNAs of Kazal-Type serine proteinase inhibitors (SPIs) were identified from hemocyte cDNA libraries of the black tiger shrimp Penaeus monodon and recognized as having 4 (SPIPm1) and 5 (SPIPm2, SPIPm3, and SPIPm4) Kazal domains. SPIPm2 encoding a complete 5-Kazal-domain inhibitor was expressed in the Escherichia coli system. Inhibitory activity of crude proteins against various serine proteases was tested using SPI activity gelatin with sodium dodecylsulfate polyacrylamide gel electrophoresis and inhibitory spectrum assays. A 32-kDa recombinant protein (rSPIPm2) showed inhibitory activity against trypsin, chymotrypsin, and subtilisin, but not elastase. Concordantly, inhibitory spectrum assays showed that crude rSPIPm2 strongly inhibited trypsin (89%) and chymotrypsin (70%), but less effectively inhibited subtilisin (8%), and did not inhibit elastase activity. Northern blot analysis of hemocyte total RNA showed 2 SPI transcripts of 1.6 and 1.7 kb in size. Tissue-specific expression using reverse transcriptase polymerase chain reaction suggests that SPIPm2 is exclusively expressed in hemocytes of P. monodon. PMID- 15756481 TI - [Domestic elder abuse and neglect--conclusions from the evaluation of a model project]. AB - The main task of a federally funded model project in the German city of Hannover was to develop approaches for prevention and intervention in the field of domestic elder abuse. Over a three year period (1998-2001), different approaches- like a telephone helpline for senior citizens, and social workers operating as counsellors for elderly people and their relatives--were tested at a local level. The paper presents results from the evaluation of the project and draws conclusions for future prevention and intervention in the field. The authors argue that the explicit use of the conceptual framework of "violence"/"abuse" creates potentials for scandalizing the issue and is therefore supportive for media appearance, whereas it can impede the approach to the main target groups (elderly people and their relatives) and reduce accessibility of counselling services for potential clients. In the light of evaluation results the focus of the project ("domestic elder abuse" or "violence against elderly people in close relationships") was too narrow for a local project. Counselling services were used in a relatively small number of cases; analyses of cases show that incidents of domestic elder abuse are often embedded in complex problem constellations. Cases brought to the attention of the model project were multifaceted and not limited to incidents of neglect and abuse of elderly care recipients caused by caregiver overload. Cases of intimate violence in partnerships and of intergenerational violence without any of the participants being dependent on care show the need to develop a broader concept of domestic elder abuse. Integration of the concepts of domestic violence, violence against women, elder abuse/neglect and abuse/neglect in caregiving relationships is necessary on a conceptual level as well as on the level of interagency cooperation of institutions dealing with cases of "elder abuse". PMID- 15756482 TI - Factors relating to the use of physical restraints in psychogeriatric care: a paradigm for elder abuse. AB - The purpose of this study was to address one component of the complex topic "elder abuse". A prospective observational study in the psychogeriatric unit of an acute psychiatric hospital demonstrated that 30% (n=37) of all included patients (n=122) were physically restrained. The highest incidence (48%) was found in elderly patients with severe cognitive impairments (diagnosis of dementia and/or delirium) (n=60). The most commonly used devices of physical restraints were bed rails (100%), belts (trunk 93%, limbs 40%) and chair-tables ("gerichair") (41%). Most restraints occurred at the beginning of hospitalization (83%). Physical restraints were continued for many days and on average of many hours a day. Patients with low cognitive status and serious mobility impairments showed a very high risk of being restrained (p=0.015; OR 32.0 [95% CI:2.0 515.1]). Inability to perform ADL activities increased the frequency of restraint use (p=0.035; OR27.7 [95%CI: 1.3-604.1]). As possible co-factors repetitive disruptive behaviors were found. There was no significant difference between the frequency of falls in restrained or unrestrained patients during the observational period, but fall-related fractures (n=2) only occurred in restrained patients. It is possible that restraints increase the use of benzodiazepines and classical neuroleptics. These results confirm that physical restraints remain a common practice in psychogeriatric care. No evidence-based data support the value of restraints in regard to fall prevention and control of behavioral disturbances in elderly people with serious mental illness. In contrast, these devices can have serious adverse effects and mean one of the most severe interventions in fundamental human rights. PMID- 15756483 TI - Why do we use physical restraints in the elderly? AB - The use of physical restraints in the elderly is a common practice in many countries. This paper summarizes the current knowledge on the use of restraints in home care, hospitals and nursing homes. Between 1999-2004 the reported prevalence numbers range from 41-64% in nursing homes and 33-68% in hospitals; numbers of restraint use in home care are unknown. Bed rails and belts have been reported as the most frequently used restraints in bed; chairs with a table and belts are the most frequently reported restraints in a chair. It is evident that physical restraints in most cases are used as safety measures; the main reason is the prevention of falls. In the hospital setting, the safe use of medical devices is also an important reason for restraint use. Predictors for the use of physical restraints are poor mobility, impaired cognitive status and high dependency of the elderly patient and the risk of falls in the nurses' opinion. Furthermore, there are indications that restraint use is related to organizational characteristics. Finally, many adverse effects of restraint use have been reported in the literature, like falls, pressure sores, depression, aggression, and death. Because of the adverse effects of restraints and the growing evidence that physical restraints are no adequate measure for the prevention of falls, measures for the reduction of physical restraints are discussed and recommendations are made for future research. PMID- 15756484 TI - [Causes of death in nursing homes--a retrospective autopsy study]. AB - Autopsies from 34 fatalities in nursing homes covering a 10-year period (1991 2000) were retrospectively analyzed with regard to criminal investigative issues, causes of death and relevance of the forensic medical findings for the outcome of the criminal investigations. The material consisted of 22 females and 12 males, with the ages ranging from 62 to 102. The allegations to be investigated comprised wrongful death, improper nursing, medical maltreatment, suspicion of poisoning and the causal relationship between falls and the occurrence of death. The manner of death, respectively cause of death encountered included 21 deaths from natural cause (coronary insufficiency, pneumonia), 9 accidental deaths (hypothermia, choking, scalding burns and injuries due to fall) and 4 cases which remained undetermined. In 30 out of 34 cases, the cause of death was unequivocally established. In every case, the medicolegal investigation provided adequate evidence for the legal assessment. Of particular significance was the exclusion of a causal relationship between falls and the occurrence of death. Without exception, all investigations failed to uncover criminal causes for death in this sample. PMID- 15756485 TI - [Effectiveness of a multifactorial intervention to reduce physical restraints in nursing home residents with dementia]. AB - At present, observational studies and expert opinion are the best evidence for the use of physical restraints. Large regional and national disparities are described in acute and long-term care. Epidemiological data demonstrate a prevalence of 3-5% body-fixed or near body restraint devices. The hip fracture rate in Germany are approximately 50 per 1000 resident years. Between 40-50% of the residents in nursing homes are treated with psycho-tropic medication potentially limiting their physical mobility. The presented study protocol was designed to test the effectiveness of a multifactorial intervention to reduce physical restraints in long-term care (LTC) residents particularly with cognitive impairment. The intervention consists of an educational and an organizational part to empower staff members to improve their skills and practice in using restraints. Technical devices to reduce fall related injuries are additionally offered to the LTC facilities. The study population includes 200 LTC residents in 54 facilities in three states in Germany. The sample size calculation was based upon a 5% prevalence rate in the control group and an expected reduction of 50% in the intervention group. The protocol is a waiting-list control design. All waiting facilities will be offered to participate after their waiting period. Primary endpoints are the number of restrained residents and resident time (hours) of being restrained. The use of psychotropics, falls, fall-related injuries and the incidence of residents newly being restrained is being monitored. The study starts with a baseline documentation of all facilities followed by randomization and a three month intervention. Change agents will be responsible for the intervention. Technical devices will include a newly developed soft hip protector and sensor mats which notice the intent of leaving the bed. The aim of the study is to develop an evidence-based model for a knowledge transfer project to implement minimum restraint environments in LTC. PMID- 15756486 TI - [Training of communication skills in stationary long care homes--the evaluation of a model project to develop communication skills and transfer it into practice]. AB - The training of communication skills of professional caregivers in six homes for elderly people has been developed and evaluated in a model project. The purpose of the project was to strengthen the staff's orientation towards the residents, their needs, handicaps and abilities. Therefore, a series of 8 in-house training courses as well as procedures to establish the contents of the program into daily care-giving (transfer) have been developed and implemented with six teams during one year. The evaluation included interviews, questionnaires and observations and was realized with participants and non-participants of the program once before the implementation of the training program and once afterwards. We found evidence for positive effects of the training: although the staff's positive self perception of the climate of communication remained stable and mainly not affected by the training, this was contradictory to the observations. The way of giving information to the residents was improved by the training program as well as the quality of relations between staff and residents. Again, sending messages about oneself which are not care-oriented had not been affected by the training as to the observations of care giving situations. Although the staff's self perception about the change of sending these messages was highly positive.Thus, the further development of the training program has to consider these effects.To establish the transfer of the training program into daily care giving, it proved to be helpful to specify exercises after each session which had to be carried out and discussed by the participants until the next training session. PMID- 15756487 TI - [Substance abuse in middle and old age-everyday drug alcohol and nicotine: use and abuse]. AB - Alcohol as a semi-luxury item is socially accepted in our society and its usage constitutes a normative behaviour. The transition from consumption to abuse is often rapid. Results supply evidence for widely spread alcohol addictions in Germany. The following paper examines the consumption and the abuse of alcohol using the data from the Interdisciplinary Longitudinal Study of Adulthood (ILSE). The findings of a questionnaire investigating diet and medical examination data [interview data referring to alcohol consumption and laboratory findings concerning Gamma-Glutamyl-Transferase levels (Gamma-GT-levels)] based on two cohorts (years of birth 1930-1932 and 1950-1952) from centres of investigation in Heidelberg and Leipzig are adopted in this study. The centre comparison indicates higher alcohol consumption in the Leipziger study participants. The findings register less alcohol usage at the second measurement point, where as the Gamma GT-levels rise and a greater endangering of health can, therefore, be noted. Besides alcohol, cigarettes are a widely accepted everyday drug in Germany. The health risks are enormous (e. g. high blood pressure, bronchitis and lung cancer). The smoking habits of the two cohorts will be described at two measurement points concerning participants from Heidelberg and Leipzig. The cohort comparison demonstrates a significant disadvantage of the younger women, although, as for as, the men are concerned no clear difference can be found. PMID- 15756488 TI - Geriatric care at the EU level? How to join? PMID- 15756492 TI - [The journal: on one's own account]. PMID- 15756493 TI - [Interdisciplinary rheumatology in internal medicine]. PMID- 15756494 TI - [Rheumatic and hemato-/oncological disorders]. AB - The association of rheumatic diseases with common and some of the less common hematological features such as different forms of anemia, leukopenia and thrombopenia are described in this article. In addition, the occurrence of malignancy in rheumatoid arthritis, systemic lupus erythematosus, myositis and scleroderma and possible causes are discussed. On the other hand, this review also focuses on various rheumatological manifestations of hematological diseases such as leukemia and lymphoma. The aim of the article is to give an overview of the various associations between rheumatological and hemato-/oncological diseases that have to be considered in clinical practice. PMID- 15756495 TI - [Cardiovascular manifestations in inflammatory rheumatic diseases]. AB - First descriptions about cardiovascular manifestations (mostly myocarditis and endocarditis) in inflammatory rheumatic diseases were dated at the end of the 19(th) century. Inflammatory rheumatic diseases show an increased cardiovascular manifestation and mortality, but the reasons for this are unknown. This overview presents the current knowledge about cardiovascular manifestations in different inflammatory rheumatic diseases for an efficient differential diagnostic and clarification in the daily practice. PMID- 15756497 TI - [Liver and rheumatism]. AB - There are multiple relations between rheumatic diseases and the liver, nevertheless the liver is extremely rare involved in rheumatic diseases. "Elevated liver enzymes" are quite often found in patients who are under the medication with immunosuppressive drugs or/and non-steroidal antirheumatics. The most frequent cause for "elevated liver enzymes" are toxic and allergic side effects of drugs; however, in rare cases it might be extremely helpful to examine, whether an independent liver disease exists. Underlying liver diseases which might be associated with the rheumatic disorder or exist accidentally may change the therapeutic management of the patient. If the liver disease present can cause the rheumatic disorder (e. g. virus-induced vasculitis, hemochromatosis), a specific hepatological therapy should precede the immunosuppression. PMID- 15756496 TI - [Renal manifestations of rheumatic diseases]. AB - Inflammatory rheumatic diseases are frequently complicated by subclinical or overt renal manifestations. This is well known for the connective tissue diseases and vasculitides in which renal disease can be of significant prognostic value and therapeutic implication. However, rheumatoid arthritis and the spondyloarthritides can also be associated with direct renal manifestation or with secondary renal AA-amyloidosis. The clinician should be aware of the different glomerular (i. e. nephritic or nephritic syndrome, rapidly progressive renal disease) and tubulo-interstitial syndromes. In any case of renal dysfunction in a rheumatic patient, the differential diagnosis should include renal disease independent from the rheumatic condition, infection, and drug induced renal toxicity. PMID- 15756499 TI - [Selective inhibition of T-cell activation with fusion protein CTLA4lg as a new treatment option for rheumatoid arthritis]. PMID- 15756498 TI - [Large vessel vasculitis as cause of fever of unknown origin (FUO) or systemic inflammation. Diagnosis using 18-F-fluor-2-deoxy-D-glucose positron emission tomography ((18)F-FDG-PET)]. AB - BACKGROUND: Diagnosis and treatment of FUO or systemic inflammation with unknown reason are still a great challenge for the treating physician. We used (18)F-FDG PET for further diagnostic work in patients in whom a diagnosis could not be established despite intensive diagnostic efforts. METHODS/RESULTS: We studied nine patients with (18)F-FDG-PET. Two female patients with known Takayasu's arteritis but undefined disease activity, and seven patients with the clinical suspicion of an underlying large vessel vasculitis. The diagnosis of active vasculitis could be confirmed by the PET-results in eight patients. Active vasculitis could be nearly ruled out in one. The diagnoses could be confirmed by follow-up visits. CONCLUSION: (18)F-FDG-PET is a useful diagnostic tool in patients with unclear systemic inflammation and FUO. Especially when large vessel vasculitis is suspected, further diagnostic work by PET seems to be of benefit. Furthermore, it offers the opportunity to evaluate disease activity and to check which vessels are involved. PMID- 15756500 TI - [What's new in infantile and adolescent rheumatology? Report of the 13th Anniversary of the Working Group on Infantile and Adolescent Rheumatology in Halle from 14 to 16 November 2003]. PMID- 15756501 TI - [Rheumatism and heart: autoimmune mechanisms link rheumatic to cardiovascular diseases]. PMID- 15756502 TI - [The magician from Paris (Jean Martin Charcot 1825-1893). II. Artist and man with a contradiction]. PMID- 15756503 TI - [Changes for rheumatology in the G-DRG system 2005]. AB - The German prospective payment system G-DRG has been recently adapted and recalculated. Apart from the adjustments of the G-DRG classification system itself changes in the legal framework like the extension of the "convergence period" or the limitation of budget loss due to DRG introduction have to be considered. Especially the introduction of new procedure codes (OPS) describing the specialized and complex rheumatologic treatment of inpatients might be of significant importance. Even though these procedures will not yet develop influence on the grouping process in 2005, it will enable a more accurate description of the efforts of acute-rheumatologic treatment which can be used for further adaptations of the DRG algorithm. Numerous newly introduced additive payment components (ZE) result in a more adequate description of the "DRG products". Although not increasing the individual hospital budget, these additive payments contribute to more transparency of high cost services and can be addressed separately from the DRG-budget. Furthermore a lot of other relevant changes to the G-DRG catalogue, the classification systems ICD-10-GM and OPS-301 and the German Coding Standards (DKR) are presented. PMID- 15756505 TI - [Professor Dr. of Medicine Erika Gromnica-Ihle on her 65th birthday]. PMID- 15756511 TI - Reactive and proactive aggression in children--a review of theory, findings and the relevance for child and adolescent psychiatry. AB - The clinical population of aggressive children diagnosed as having an oppositional defiant disorder (ODD) or a conduct disorder (CD) is heterogeneous, both with respect to behaviour and aetiology. Recently, the following distinction has been proposed that might further clarify this heterogeneity: reactive aggression is an aggressive response to a perceived threat or provocation, whereas proactive aggression is defined as behaviour that anticipates a reward. In this article we examine various aspects of this distinction. We will [1] examine the evidence that reactive and proactive aggression are distinct phenomena by discussing the theories underlying the distinction between the subtypes in humans and we briefly review evidence for a similar distinction in animals; [2] we critically review the literature on the measurement in children via questionnaires and behavioural observations; we then point out that the correlation observed between the subtypes is due to the fact that many children show both types of aggression; [3] we review the literature on specific characteristics of the subtypes giving attention to social information processing, peer status, biological correlates and developmental history, and demonstrate that there is some evidence to suggest that reactive and proactive aggression are distinct dimensions; [4] we discuss the relevance of the distinction between reactive and proactive aggression for child and adolescent psychiatry. PMID- 15756510 TI - The scientific foundation for understanding attention-deficit/hyperactivity disorder as a valid psychiatric disorder. AB - Continued questioning of the validity of a diagnosis of attention deficit/hyperactivity disorder (ADHD) has created uncertainties about its management in the minds of many clinicians and the public. Inaccurate beliefs about the validity of ADHD hinder the clinical care of many ADHD patients and lead to confusion about the need to seek out or accept treatment. Critics describe ADHD as a diagnosis used to label difficult children who are not ill but whose behavior is at the extreme end of normal. They further contend that, far from having a biological basis, ADHD results from poor parenting and ineffective teaching practices. Such attitudes do much to further stigmatize patients and their families and increase the burden of this debilitating condition. This review attempts to address these challenges by presenting evidence to show that ADHD meets the criteria for a valid psychiatric diagnosis. Not only does it cause specific disabling symptoms that frequently persist into adulthood, but many studies show it has a biological basis and a characteristic response to treatment. Such data support the idea that ADHD is a valid diagnostic category. PMID- 15756512 TI - Correspondence of parent and teacher reports in medication trials. AB - Although much has been written about the agreement of parents and teachers regarding the diagnosis of attention-deficit/hyperactivity disorder (ADHD), some uncertainty remains about their agreement when assessing change during drug treatment. To address this issue, we analyzed data from a placebo-controlled short-term study and an open-label long-term study of ADHD children treated with OROS methylphenidate (MPH). Both reporters agreed that OROS MPH was efficacious for symptoms of inattention/overactivity and oppositionality/defiance and that the effect was greater for inattention/overactivity than oppositionality/defiance. Thus, in clinical trials, having two reports may not be absolutely essential for assessing the efficacy of ADHD medications. We also computed diagnostic efficiency statistics and found a high probability that parents would confirm teacher reports of therapeutic improvement and a somewhat lower probability that teachers would confirm parent reports of therapeutic improvement. In contrast, neither reporter was likely to confirm the other reporter's report of no improvement or worsening. Thus, when given a report of no improvement from one setting, clinicians cannot be certain about clinical status in the other setting. For symptoms of oppositionality and defiance, we also saw that parent- teacher agreement about improvement was better than agreement about lack of improvement. However, for these symptoms, teacher reports were somewhat better predictors of parent reports than vice-versa. PMID- 15756513 TI - Russian child mental health--a cross-sectional study of prevalence and risk factors. AB - BACKGROUND: The fall of communism and subsequent economic crises have been followed by major social and health problems. High rates of child mental health problems are frequently cited by the Russian media, though there is little relevant evidence. AIMS: The aim of this study was to investigate the prevalence and associations of child mental health problems in Russia using internationally recognised measures and diagnostic systems. METHOD: A two-stage, two-phase cross sectional survey of the mental health of 7- to 14-year-olds involved random sampling of schools, followed by random sampling of pupils from school lists. A sample of 448 children was obtained, representing an 83% participation rate. In the first phase, screening measures of psychopathology and risk were administered to parents, teachers and 11- to 14-year-olds. In the second phase, more detailed psychiatric assessments were carried out for subgroups of screen-positive and screen-negative children (N=172). RESULTS: The prevalence of psychiatric disorder was about 70% higher than that recently found in Britain with comparable measures, but there were few differences between Britain and Russia in type of disorder or key risk factors. CONCLUSION: There is a pressing need for evidence based mental health treatments to be made widely available to Russian children and adolescents. PMID- 15756514 TI - The use of clonidine for severe and intractable sleep problems in children with neurodevelopmental disorders--a case series. AB - This paper reports on the use of clonidine for the treatment of severe sleep problems associated with behavioural difficulties in children with neurodevelopmental disabilities. Data were obtained from reviewing the case notes of a series of six children with neurodevelopmental disorders of different nature and severity, presenting with problematic sleep. All children in this group showed maintained improvements in their sleep pattern following the use of clonidine with only mild side-effects reported. PMID- 15756516 TI - Transient eating disorder in early childhood--a case report. AB - Although children in infancy present eating problems of multifarious nature, the diagnostic classification of infant eating disorders remains markedly deficient. The authors present a case exhibiting transient eating disorder in early childhood, alongside discussion of some considerations relevant to this age group. The subject was a boy aged 5 years 6 months at first presentation. Starting with an inability to swallow "sushi" at dinner, he was brought to the clinic for inability to ingest food or liquids. Treatment was planned as play therapy for the patient, and psychological interviews for the mother. The child's symptoms were improved in five sessions, but interviews of the mother revealed eating disorders such as bulimia and vomiting, and sexual problems in the marital relationship, necessitating long-term therapy. PMID- 15756515 TI - The effects of prenatal stress on temperament and problem behavior of 27-month old toddlers. AB - AIM: To examine, in a prospective study, the influence of prenatal stress on infant temperament and problem behavior. METHOD: Self-report data on stress and anxiety, and levels of cortisol in saliva were collected from nulli-parous women during pregnancy. Temperament of the child was measured at 27 months by parent report on the Infant Characteristics Questionnaire. Behavior of the child was assessed by direct observation during the administration of the Bayley Scales of Development 2-30, and by parent report on the Child Behavior Checklist 2-3. RESULTS: Complete data were available for 103 healthy toddlers. Logistic regression analyses were performed and results were adjusted for possible prenatal, perinatal and postnatal confounders. Perceived stress during pregnancy was a predictor of lower levels of restless/disruptive temperament (OR=0.77), more total behavioral problems (OR=1.17), and more externalizing behavioral problems (OR=1.12) in 2-year-olds. Fear of bearing a handicapped child was a predictor of higher levels of restless/disruptive temperament (OR=1.39) and more attention regulation problems in toddlers (OR=1.46). CONCLUSIONS: Increased levels of maternal prenatal stress appear to be associated with temperamental and behavioral problems in toddlers. PMID- 15756518 TI - Cognitive effects of chemotherapy and/or cranial irradiation in adults. AB - BACKGROUND: Cognitive effects after cranial radiotherapy are widely discussed, but there is growing evidence that chemotherapy may also induce changes in neuropsychological functioning. This review summarizes the published literature regarding cognitive functioning after cancer therapy in adult patients. MATERIAL AND METHODS: 63 reports from January 1980 to July 2003 assessing objective cognitive effects of irradiation and/or chemotherapy by neuropsychologic evaluation were analyzed. 57 studies with 3,424 patients were included for evaluation. RESULTS: The results of this review confirm that both chemotherapy and irradiation can result in cognitive deficits. No clinically relevant differences are found for cognitive deficits, cognitive impairment rate, and single cognitive domains, when chemotherapy, cranial irradiation and combined radio- and chemotherapy were compared. Only 28 trials with 1,000 patients report quantitative data on patients with cognitive deficits after therapy. There are 44.1% (range 18-75%) of 451 patients in the chemotherapy group, 44.0% (range 29 83%) of 320 patients in the radiotherapy group, and 64.5% (range 30-100%) of 229 patients in the combined irradiation and chemotherapy group with cognitive deficits. Furthermore, cognitive functioning below average before chemo- or radiotherapy is found in subgroups of cancer patients. CONCLUSION: There is evidence of cognitive impairment in adult tumor patients after chemotherapy similar to effects after cranial irradiation. Cognitive functioning below average before therapy may be due to paraneoplastic effects. More prospective studies with a long-term follow-up using standardized neuropsychometric testing, assessment of premorbid intelligence, and suited control groups are needed. PMID- 15756519 TI - Efficacy and toxicity of postoperative temozolomide radiochemotherapy in malignant glioma. AB - PURPOSE: To evaluate the feasibility, safety and efficacy of daily temozolomide concurrent with postoperative radiotherapy in malignant glioma. PATIENTS AND METHODS: From 11/1999 to 03/2003, n = 81 patients aged 15-72 years (median 52 years, Karnofsky score 80-100% in 83%) suffering from primary glioblastoma (n = 47), anaplastic astrocytoma (n = 6), anaplastic oligodendroglioma (n = 16), and recurrent glioma (n = 12) were treated. Patients with primary gliomas received a combination of postoperative radiotherapy (60 Gy/1.8- to 2.0-Gy fractions) and daily oral temozolomide (75 mg/m(2)) at all irradiation days (30-33 doses), while recurrent tumors were treated with 45-60 Gy and temozolomide. Initially, 6/81 patients had daily temozolomide doses of 50 mg/m(2). RESULTS: In total, 70/81 patients (86%) completed both radio- and chemotherapy. Grade 1 nausea/vomiting was seen in 28%, grade 2 in 11%, grade 3 in 1%. Antiemetics were applied in 41%. Hematologic toxicities were observed as follows: leukopenia grade 3/4 1%, lymphopenia grade 3/4 46%, thrombopenia grade 3/4 1%. Two patients under dexamethasone suffered herpes encephalitis after one and 16 doses of temozolomide (75 mg/m(2)). Median survival was 15 months for glioblastoma. In oligodendroglioma patients, a 4-year survival rate of 78% was observed. CONCLUSION: Postoperative radiochemotherapy with 30-33 daily doses of temozolomide (75 mg/m(2)) is safe in patients with malignant glioma. The combined schedule is effective in oligodendroglioma patients and may prolong survival in glioblastoma. Effort should be taken to minimize corticosteroid doses, since both steroids and temozolomide lead to immunosuppression. PMID- 15756520 TI - Expression of p53 and p21 in primary glioblastomas. AB - BACKGROUND AND PURPOSE: Primary glioblastomas (GBMs) are highly radioresistant, and in contrast to secondary GBMs, they bear wild-type (wt) p53 protein, which is stabilized in a proportion of these tumors. Therefore, it was investigated in vivo whether p53 expression has prognostic value in patients undergoing radiochemotherapy. Additionally, the authors tried to identify, in vitro, subgroups of primary GBM with different susceptibilities to irradiation, on the basis of their p53 and p21 responses to ionizing radiation. MATERIAL AND METHODS: Tumor tissue samples from 31 patients suffering from primary GBM undergoing a combined radiochemotherapy with topotecan were investigated. The percentage of cells expressing p53 protein was determined immunohistochemically. Additionally, primary cultures from eleven primary GBMs were established and investigated. p53 and p21 expressions were evaluated before irradiation with 10 Gy and at 2 and 8 h after irradiation. p53 protein expression was measured by Western analysis and p21 mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The percentage of p53-positive cells within the tumor specimens obtained from the 31 patients ranged from 0% to 28%, the median value being 4.3%. No significant correlation with disease-free survival or overall survival was found. In vitro, p53 protein was detected in seven of eleven cultures from primary GBM. After irradiation a decrease in p53 protein expression was seen in six of the seven p53-positive cultures. Half of the cultures (two of four) without basal p53 expression showed an increase in p53 expression after irradiation. Basal overexpression of p21 was detected in six of the eleven cultures; in four out of six irradiation led to a decrease in p21 expression. In all cell lines (five of eleven) initially showing absent p21 expression, irradiation induced p21 expression. Despite these responses, G1 arrest was not detectable in any of the GBM cultures. CONCLUSION: p53 protein expression in vivo does not correlate with the outcome of patients with primary GBM. Therefore, p53 protein content per se does not appear to be a helpful prognostic factor for prognosis-adapted therapy in primary GBM. By contrast, primary GBM cells in vitro show different and independent responses in their p53 and p21 pathways to ionizing radiation. The failure of G1 arrest seems to be due to a functional defect in the p53 pathway, either because p21 was not induced or because of an unidentified defect downstream from p21. PMID- 15756521 TI - Dose to bone marrow using IMRT techniques in prostate cancer patients. AB - PURPOSE: To investigate the dose distribution in active bone marrow of patients undergoing intensity-modulated radiotherapy (IMRT) for prostate cancer and compare it to the distribution in the same patients, if they had been treated using conformal plans, in order to develop criteria for optimization to minimize the estimated risk of secondary leukemia. PATIENTS AND METHODS: Mean bone marrow doses were calculated for ten patients with localized prostate cancer who underwent whole-pelvis IMRT and compared to three-dimensional conformal (3-D CRT) plans prepared for the same patients. Also for comparison, the IMRT and 3-D CRT plans were produced to simulate the treatment of the prostate gland only. To measure the dose to extrapelvic bone marrow, three thermoluminescent diode (TLD) chips were placed in the middle of the sternum region inside the Rando phantom. RESULTS: For both the pelvic and prostate-only volumes, the IMRT plans were superior to 3-D CRT plans in reducing the high dose volume to the rectum, the bladder and the small bowel while maintaining acceptable coverage of the planning target volume (PTV). For the pelvic treatment group the IMRT plans, compared to 3 D CRT, reduced the high dose volume (> 20 Gy) to os coxae, which is the main contributor of dose to pelvic bone marrow, but increased the middle dose volume (10-20 Gy). No statistically significant differences were observed for lower dose volumes (< 5 Gy). For the prostate-only treatment the IMRT plan increased the high dose volume and slightly decreased the low dose volume of pelvic bone marrow. However, for both treatments the leakage dose to extrapelvic sites was higher by a factor of 2 in IMRT plans. CONCLUSION: There are significant differences in the dose-volume histograms of bone marrow doses from 3-D CRT and from IMRT. Pronounced dose inhomogeneity reduces the risk of leukemia compared to homogeneous radiation exposure of the bone marrow. The mean bone marrow dose is therefore not a useful criterion to judge plan quality, since scattered low doses to distant sites may be more critical than the high dose volumes receiving > 10 Gy. The number of monitor units needed to deliver an IMRT plan affects leakage dose and their incorporation into planning constraints should be considered. PMID- 15756522 TI - Testicular dose in prostate cancer radiotherapy: impact on impairment of fertility and hormonal function. AB - PURPOSE: To determine the dose received by the unshielded testicles during a course of 20-MV conventional external-beam radiotherapy for patients with localized prostate cancer. Critical evaluation of the potential impact on fertility and hormonal impairment in these patients according to the literature. PATIENTS AND METHODS: The absolute dose received by the testicles of 20 randomly selected patients undergoing radiotherapy of prostate cancer was measured by on line thermoluminescence dosimetry. Patients were treated in supine position with an immobilization cushion under their knees. A flexible tube, containing three calibrated thermoluminescence dosimeters (TLDs) was placed on top or underneath the testicle closest to the perineal region with a day-to-day alternation. The single dose to the planning target volume was 1.8 Gy. Ten subsequent testicle measurements were performed on each patient. The individual TLDs were then read out and the total absorbed dose was calculated. RESULTS: The mean total dose (+/- standard deviation) measured in a series of 10 subsequent treatment days in all patients was 49 cGy (+/- 36 cGy). The calculated projected doses made on a standard series of 40 fractions of external-beam radiotherapy were 196 cGy (+/- 145 cGy). The results of this study are appraised with the available data in the literature. CONCLUSION: The dose received by the unshielded testes can be assessed as a risk for permanent infertility and impairment of hormonal function in prostate cancer patients treated with external-beam radiotherapy. PMID- 15756523 TI - Can intensity-modulated radiation therapy of the paraaortic region overcome the problems of critical organ tolerance? AB - BACKGROUND AND PURPOSE: The recent RTOG guidelines for future clinical developments in gynecologic malignancies included the investigation of dose escalation in the paraaortic (PO) region which is, however, very difficult to target due to the presence of critical organs such as kidneys, liver, spinal cord, and digestive structures. The aim of this study was to investigate intensity-modulated radiotherapy's (IMRT) possibilites of either increasing, in a safe way, the dose to 50-60 Gy in case of macroscopic disease or decreasing the dose to organs at risk (OR) when treatment is given in an adjuvant setting. MATERIAL AND METHODS: The dosimetric charts of 14 patients irradiated to the PO region at the Department of Radiation Oncology, University Hospital of Liege, Belgium, in 2000 were analyzed in order to compare six-field conformal external beam radiotherapy (CEBR) and five-beam IMRT approaches. Both CEBR and IMRT investigations were planned to theoretically deliver 60 Gy to the PO region in the safest way possible. Dose-volume histograms (DVHs) were calculated for clinical target volume (CTV), planning target volume (PTV), and OR. Student's t test was used to compare the paired DVH data issued from CEBR and IMRT planning. RESULTS: The IMRT approach allowed to cover the PTV at a higher level as compared to CEBR. Using IMRT, the maximal dose to the spinal cord was reduced from 42.5 Gy to 26.2 Gy in comparison with CEBR (p < 0.00001). Doses to the kidneys were significantly reduced, with < 20% receiving >or= 20 Gy in the IMRT approach (p < 0.00001). Irradiation of digestive structures was not different, with < 25% receiving 35 Gy. Doses to the liver remained low regardless of the method used. CONCLUSION: At 60 Gy, IMRT is largely sparing the spinal cord and kidneys as compared to CEBR and represents an interesting approach not only for dose escalation up to 50-60 Gy (probably facilitating the radiochemotherapy approaches) but also in an adjuvant setting at lower doses. The dosimetric data of this study are in the same range as those published recently with a dynamic arc conformal approach. PMID- 15756524 TI - Selective radioprotection of normal tissues by Bowman-birk proteinase inhibitor (BBI) in mice. AB - BACKGROUND AND PURPOSE: The efficacy of radiotherapy is limited by the response of normal tissues within the radiation field. The application of normal-tissue specific radioprotectors may improve the therapeutic benefit of radiotherapy. The purpose of the present study was to explore the in vivo normal-tissue radioprotective potential of Bowman-Birk proteinase inhibitor (BBI), which acts as a normal-cell-specific radioprotector in vitro. MATERIAL AND METHODS: The leg contracture assay in mice, a model system assessing radiation-induced fibrotic processes, was used. To determine whether BBI acts also as a radioprotector of tumors (i. e., FSA and FSAII), the tumor growth delay assay was used. RESULTS: Radiation induced leg contracture in mice with a maximum of about 8 mm at day 150 after irradiation. Treatment of mice with 100 mg/kg BBI before irradiation reduced leg contracture by > 4 mm, by about 50% (p < 0.05, t-test). Doses < 100 mg/kg were ineffective, and doses > 100 mg/kg did not further increase the degree of radioprotection. By contrast, BBI did not induce radioprotection of either TP53-mutated FSA or TP53-normal FSAII tumor xenografts in mice, which argues for normal-tissue-specific effect. CONCLUSION: BBI acts as a potent selective normal tissue radioprotector in vitro and in vivo, apparently without protecting tumors, and thus has the potential to improve clinical radiotherapy. PMID- 15756525 TI - Radiosensitivity of tumor cell lines after pretreatment with the EGFR tyrosine kinase inhibitor ZD1839 (Iressa). AB - BACKGROUND AND PURPOSE: The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor ZD1839 (Iressa) reduces survival and augments radiation response of certain tumor cells. The aim of this study was to identify cellular events that are associated with the modulation of radiosensitivity by ZD1839. MATERIAL AND METHODS: Three tumor cell lines (A549, H596, FaDu) were exposed to ionizing radiation, treatment with ZD1839, and combined treatment. Clonogenic cell survival was determined by colony assays, EGFR and transforming growth factor (TGF-)alpha expression by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), cell cycle distribution and apoptosis by flow cytometry. RESULTS: In A549 and H596 cells ZD1839 had little effect on clonogenic growth, but survival curves revealed a radiosensitizing effect of 5 microM ZD1839 on A549 cells. Both cell lines expressed moderate amounts of EGFR mRNA and very low levels of TGF-alpha mRNA. FaDu cells expressed relatively high amounts of EGFR and TGF-alpha transcripts and showed marked inhibition of clonogenic growth, reduction of S-phase cells, and induction of apoptosis after treatment with 1 microM ZD1839 and combined treatment. Surprisingly, the subpopulation of FaDu cells surviving ZD1839 pretreatment was more radioresistant. Exposure to ZD1839 caused a decrease in EGFR mRNA expression in A549 cells, no change in H596, and even an increase in FaDu cells. CONCLUSION: The sensitivity to ZD1839 correlated with the EGFR expression level, an inhibition of cell proliferation, and induction of apoptosis in the cell lines analyzed. A radiosensitizing effect of ZD1839 was associated with downregulation of EGFR mRNA expression. PMID- 15756527 TI - [Solitary osteochondroma of the scapula. A rare differential diagnosis with unspecific shoulder pain]. AB - Solitary osteochondroma of the scapula is rare. Because of the atypical location with unspecific shoulder pain, the diagnosis is often made late. We present a 35 year old patient with a solitary ostochondroma of the scapula with unspecific pain of the shoulder-neck region over about 4 years. In a thorax x-ray 2 years ago, a tumor like lesion was found. CT and MRI scan led to suspicion of a solitary osteochondroma of the scapula. Because of progressive thoracic pressure pain, a wide excision was made. The problems of diagnosis and therapy are presented and the literature is discussed. PMID- 15756526 TI - [A bilaterally dislocated intraarticular calcaneus fracture in a 13-year old boy]. AB - Bilaterally dislocated intraarticular calcaneus fractures in childhood are rare. This case report shows that adequate preoperative tissue management, open anatomical reduction, and lateral plate osteosynthesis by an experienced surgeon according to the principles of adult surgery achieve a good functional result. PMID- 15756529 TI - [Obsessive compulsive disorder and schizophrenia. An attempt at psychopathologic differentiation by focusing on the single phenomenon of obsession]. AB - This paper follows the hypothesis that differentiation between obsessive compulsive disorder and schizophrenia is possible only by focusing on the single phenomenon of obsession. The declaration of a nosological specificity of obsession is set against the current view of ICD-10 and DSM-IV, of obsession as a ubiquitous nonspecific phenomenon appearing with comorbidities. The historic development of these two most different views of obsession and their combination is explained. The distinction between obsessive compulsive disorder and schizophrenia can be made at the psychopathologic dividing line between obsession and delusional thoughts. Examination of the literature on the transition from obsessive compulsive disorder into schizophrenia shows that there is no clear link between obsession and schizophrenia. PMID- 15756528 TI - [Complex pelvic trauma caused by an accidental side split]. AB - Complex pelvic ring fractures are defined as injuries of the pelvic ring in association with lesions of the pelvic organs or the pelvic soft tissues. These injuries are typically caused by high-energy accidents. In contrast to the typical mechanism of injury a case is described in which a low energy trauma led to a complex pelvic ring trauma. An obese woman suffered an open-book injury of the pelvis with severe open urogenital soft-tissue damage by accidentally doing forced splits. Primary stabilization of the pelvic ring with external fixation and secondary internal fixation with a double-plate osteosynthesis of the symphysis led to a good clinical outcome concerning the osseous lesion. The urogenital injuries with rupture of the bladder, the urethra and the vagina led despite immediate urological management to an incontinence, which finally required definitive urine drainage via an ileum conduit. PMID- 15756530 TI - [Long-acting methylphenidate. An alternative medical therapy for adult patients with attention deficit hyperactivity disorder]. AB - We report a 44-year-old female patient with attention deficit hyperactivity disorder (ADHD), combined subtype (DSM-IV: 314.01), who was treated with 0.5 mg of short-acting immediate-release methylphenidate/kg body weight given t.i.d. (total daily MPH IR dosage 45 mg). Under this medication, the patient reported significant reduction of symptoms. However, several times a day she experienced severe rebound phenomena with pronounced concentration disturbances, unrest, and dysphoric mood. After changing the medication to long-acting methylphenidate once daily (total daily OROS MPH dosage 54 mg), the rebound phenomena stopped, with equivalent beneficial clinical effects. PMID- 15756531 TI - [From echo-sounding to color doppler sonography. The history of diagnostic ultrasonic diagnosis]. AB - This is a report concerning with the development of ultrasound imaging from the very early beginning in the late 18. century by the brothers Curie until to sophisticated imaging units, which are today used for the color coded doppler sonography. PMID- 15756532 TI - [Occupational exposure to radiation]. AB - Since the discovery of X-rays , overexposures and radiation injuries have been reported in both patients and persons exposed in the course of their professional duties. Each year, more than 2,500 million diagnostic radiological examinations, 32 million nuclear medicine examinations or treatment procedures and 5.5 million radiotherapy sessions are performed worldwide. Despite all precautions, avoidable incidents and accidents occur throughout the world every year , albeit with low frequency. Whereas diagnostic radiology is generally safe for patients and staff, interventional procedures (e.g. coronary artery dilatations) involve the risks of occupational overexposure and of skin injuries to patients. In nuclear medicine, radiation protection is focused on the introduction of new procedures with beta emitters, for example. The increasing frequency of positron emission tomography (PET) requires a special focus on shielding measures. In radiotherapy, occupational overexposure caused by malfunctions and accidents is relatively rare. PMID- 15756533 TI - [Therapy for recurrent testicular cancer]. AB - In the case of an insufficient response to primary treatment or a tumor relapse, regardless of an initially complete remission, conventional as well as high dose chemotherapy regimens are available as salvage therapy for metastatic germ cell tumors. A multimodal approach should include the radiation of simultaneously occurring brain metastases as well as the surgical resection of residual tumour masses still detectable after completion of chemotherapy. Nowadays, an attempt is made to adjust the salvage modality selected to the individual situation according to a risk stratification of patients. However, a recurrence-free survival of 50% is worse when compared with that observed after primary chemotherapy. Salvage therapy should be reserved for specialized centres due to the increased complexity of a salvage approach and a significantly increased therapy-induced morbidity. PMID- 15756534 TI - Recurrent selection with reduced herbicide rates results in the rapid evolution of herbicide resistance in Lolium rigidum. AB - There has been much debate regarding the potential for reduced rates of herbicide application to accelerate evolution of herbicide resistance. We report a series of experiments that demonstrate the potential for reduced rates of the acetyl-co enzyme A carboxylase (ACCase)-inhibiting herbicide diclofop-methyl to rapidly select for resistance in a susceptible biotype of Lolium rigidum. Thirty-six percent of individuals from the original VLR1 population survived application of 37.5 g diclofop-methyl ha(-1) (10% of the recommended field application rate). These individuals were grown to maturity and bulk-crossed to produce the VLR1 low dose-selected line VLR1 (0.1). Subsequent comparisons of the dose-response characteristics of the original and low dose-selected VLR1 lines demonstrated increased tolerance of diclofop-methyl in the selected line. Two further rounds of selection produced VLR1 lines that were resistant to field-applied rates of diclofop-methyl. The LD50 (diclofop-methyl dose required to cause 50% mortality) of the most resistant line was 56-fold greater than that of the original unselected VLR1 population, indicating very large increases in mean population survival after three cycles of selection. In vitro ACCase inhibition by diclofop acid confirmed that resistance was not due to an insensitive herbicide target site. Cross-resistance studies showed increases in resistance to four herbicides: fluazifop-P-butyl, haloxyfop-R-methyl, clethodim and imazethapyr. The potential genetic basis of the observed response and implications of reduced herbicide application rates for management of herbicide resistance are discussed. PMID- 15756535 TI - Molecular cytogenetic analysis of Brassica rapa-Brassica oleracea var. alboglabra monosomic addition lines. AB - Interspecific alien chromosome addition lines can be very useful for gene mapping and studying chromosome homoeology between closely related species. In this study we demonstrate a simple but robust manner of identifying individual C-genome chromosomes (C5, C8 and C9) in the A-genome background through the simultaneous use of 5S and 25S ribosomal probes on mitotic and meiotic chromosomes of three different Brassica rapa-B. oleracea var. alboglabra monosomic addition lines. Sequential silver staining and fluorescence in situ hybridisation indicated that 18S-5.8S-25S rRNA genes on the additional chromosome C9 are expressed in the A genome background. Meiotic behaviour of the additional chromosomes was studied in pollen mother cells at diakinesis and metaphase I. In all of the addition lines the alien chromosome was most frequently observed as a univalent. The alien chromosome C5, which carries an intercalary 5S rDNA locus, occasionally formed trivalents that involved either rDNA- or non rDNA-carrying chromosomes from the A genome. In the case of chromosomes C8 and C9, the most frequently observed intergenomic associations involved the regions occupied by 18S-5.8S-25S ribosomal RNA genes. It is possible that not all such associations represent true pairing but are remnants of nucleolar associations from the preceding interphase. Variations in the numbers and distribution of 5S and 25S rDNA sites between cultivars of B. oleracea, B. oleracea var. alboglabra and B. rapa are discussed. PMID- 15756536 TI - Combined positivity for HLA DQ2/DQ8 and IA-2 antibodies defines population at high risk of developing type 1 diabetes. AB - AIMS/HYPOTHESIS: Prevention trials in first-degree relatives of type 1 diabetic patients are hampered by large interindividual differences in progression rate to diabetes. We investigated whether specific combinations of immune and genetic markers can identify subgroups with more homogeneous progression to clinical onset. METHODS: Antibodies against islet cell cytoplasm (ICA), insulin (IAA), glutamate decarboxylase (GADA) and IA-2 protein (IA-2A) were measured in 790 non diabetic control subjects and 4,589 first-degree relatives under age 40. RESULTS: On first sampling, 11.1% of the siblings presented at least one antibody type (p<0.001 vs other relatives). During follow-up (median 52 months) 43 subjects developed type 1 diabetes (31 siblings, ten offspring of a diabetic father, two offspring of a diabetic mother). Using Kaplan-Meier survival analysis and Cox regression, IA-2A conferred the highest 5-year diabetes risk (>50%) irrespective of the number of antibodies present. In initially IA-2A-positive relatives (n=58) progression to hyperglycaemia depended more on HLA DQ status than on type of kinship (84% progression in the presence of DQ2/DQ8 vs 32% in its absence; p<0.003). In IA-2A-negative relatives (n=4,531) 5-year progression to diabetes increased with the number of other antibodies (ICA, GADA and/or IAA) (p<0.001) but overall did not exceed 10% even for two or more antibodies. Among relatives initially positive for one or more antibody type other than IA-2A (n=315), there was significantly more progression to diabetes (overall still <10%) in carriers of DQ2 (p<0.001 vs no DQ2), regardless of DQ8 status. CONCLUSIONS/INTERPRETATION: These observations suggest that the HLA-DQ-inferred risk of diabetes can proceed through two distinct pathways distinguished by IA-2A status. Combined positivity for DQ2/DQ8 and IA-2A defines a more homogeneous high-risk population for prevention trials than those used so far. PMID- 15756538 TI - Hypothyroidism in rats decreases peripheral glucose utilisation, a defect partially corrected by central leptin infusion. AB - AIMS/HYPOTHESIS: The aims of this work were to determine the effect of hypothyroidism on insulin-stimulated glucose turnover and to unravel the potential mechanisms involved in such an effect. METHODS: Hypothyroidism was induced by administration of propylthiouracil, with partial T4 substitution. Euglycaemic-hyperinsulinaemic clamps, associated with the labelled 2-deoxy-D glucose technique for measuring tissue-specific glucose utilisation, were used. To assess a possible involvement of leptin in the modulation of glucose metabolism by hypothyroidism, leptin was infused intracerebroventricularly for 6 days. A group of leptin-infused rats was treated with rT3 to determine a potential role of T3 in mediating the leptin effects. RESULTS: Compared with euthyroid rats, hypothyroid animals exhibited decreased overall glucose turnover and decreased glucose utilisation indices in skeletal muscle and adipose tissue. Leptinaemia in hypothyroid rats was lower while resistin mRNA expression in adipose tissue was higher than in euthyroid animals. Intracerebroventricular leptin infusion in hypothyroid rats partially restored overall, muscle and adipose tissue insulin-stimulated glucose utilisation and improved the reduced glycaemic response observed during insulin tolerance tests. The leptin effects were due neither to the observed increase in plasma T3 levels nor to changes in the high adipose tissue resistin expression of hypothyroid rats. The administration of leptin to hypothyroid animals was accompanied by increased expression of muscle and adipose tissue carnitine palmitoyl transferases, decreased plasma NEFA levels and reduced muscle triglyceride content. CONCLUSIONS/INTERPRETATION: Hypothyroidism is characterised by decreased insulin responsiveness, partly mediated by an exaggerated glucose-fatty acid cycle that is partly alleviated by intracerebroventricular leptin administration. PMID- 15756537 TI - Activation of imidazoline receptors in adrenal gland to lower plasma glucose in streptozotocin-induced diabetic rats. AB - AIMS/HYPOTHESIS: The present study investigated the effect of agmatine, an endogenous ligand of imidazoline receptors, on plasma glucose in streptozotocin induced diabetic rats (STZ-diabetic rats). METHODS: Plasma glucose was assessed by the glucose oxidase method. Plasma insulin and beta-endorphin-like immunoreactivity in plasma or adrenal medulla were measured by enzyme-linked immunosorbent assay. Systolic blood pressure was determined by the tail-cuff method. The mRNA levels of glucose transporter subtype 4 (GLUT4) in soleus muscle and phosphoenolpyruvate carboxykinase (PEPCK) in liver were detected by northern blotting. Protein levels of GLUT4 in soleus muscle and hepatic PEPCK were estimated using western blotting analysis. RESULTS: After intravenous injection into fasting STZ-diabetic rats for 30 min, agmatine decreased plasma glucose in a dose-dependent manner without changing systolic blood pressure. At the same time, plasma beta-endorphin-like immunoreactivity also increased in STZ-diabetic rats receiving the same treatment. Plasma glucose was significantly elevated in STZ diabetic rats by an intravenous injection of clonidine at a dose sufficient to decrease systolic blood pressure. Involvement of I(1)-imidazoline receptors and/or alpha2-adrenoceptors in this effect of agmatine was thus unlikely. The lowering of plasma glucose and increase of plasma beta-endorphin-like immunoreactivity by agmatine were abolished by pretreating the rats with BU-224 at a dose sufficient to block I(2)-imidazoline receptors. Both effects of agmatine were also abolished in adrenalectomised STZ-diabetic rats. Moreover, agmatine enhanced beta-endorphin-like immunoreactivity release from the isolated adrenal medulla of STZ-diabetic rats, an effect also blocked by BU-224. Release of beta-endorphin from the adrenal glands by I(2)-imidazoline receptor activation seems responsible for the plasma glucose-lowering action of agmatine. This was supported by the fact that intravenous injection of naloxone or naloxonazine at doses sufficient to block opioid mu-receptors inhibited the action of agmatine. In addition to lowering plasma glucose, repeated intravenous injection of agmatine into STZ-diabetic rats for 4 days also increased mRNA and protein levels of GLUT4 in soleus muscle. The same treatment also reversed the higher mRNA and protein levels of PEPCK in liver of STZ-diabetic rats. CONCLUSIONS/INTERPRETATION: Our results suggest that agmatine may activate I(2) imidazoline receptors in the adrenal gland. This enhances secretion of beta endorphin, which can activate opioid mu-receptors to increase GLUT4 gene expression and/or suppress hepatic PEPCK gene expression, resulting in a lowering of plasma glucose in diabetic rats lacking insulin. The results provide a potential new target for intervention in type 1 diabetes. PMID- 15756541 TI - The role of tumor lysis in reversible posterior leukoencephalopathy syndrome. AB - We report on a child with B-cell lymphoma who developed hypertension and reversible posterior leukoencephalopathy syndrome (RPLS) after chemotherapy conducted during recovery from tumor lysis syndrome. After recovery from RPLS, the patient received further combination chemotherapy without recurrence of the neurological signs or symptoms suggestive of RPLS. Many etiological factors have been reported in the development of RPLS; however, little attention has been paid to tumor lysis syndrome as a contributory factor for RPLS. Tumor lysis syndrome can precipitate the development of RPLS in patients with hematological malignancies who are undergoing chemotherapy. Knowledge and awareness would help facilitate immediate management such as normalization of blood pressure and temporary cessation of chemotherapy, helping to avoid irreversible brain damage. PMID- 15756540 TI - Vascular damage induced by type 2 diabetes mellitus as a risk factor for benign prostatic hyperplasia. AB - AIMS/HYPOTHESIS: The aim of this study was to evaluate the relationship between benign prostatic hyperplasia (BPH) and arteriosclerosis shown in a model of type 2 diabetes in a trans-sectional population study using contrast-enhanced colour Doppler ultrasound for exact assessment of prostatic blood flow. METHODS: Contrast-enhanced transrectal colour Doppler ultrasound was performed using a microbubble-based ultrasound enhancer SonoVue for evaluating prostate vascularity (transitional zone [TZ] and peripheral zone [PZ]) in diabetic BPH patients, non diabetic BPH patients and healthy subjects. Computer-assisted quantification of colour pixel intensity (CPI) was used to objectively evaluate the prostate vascularity. Resistive index measurements were obtained in the TZ and the PZ. Findings were compared between these three groups. RESULTS: TZ-CPI was significantly lower in diabetic patients than in non-diabetic BPH men (p=0.001), whereas the CPI of the PZ showed no difference between these two groups (p=0.978). TZ-CPI of patients with diabetic and non-diabetic BPH were significantly lower than in controls (p<0.001), but no difference was found between diabetic and healthy patients in the PZ (p=0.022) and borderline significance was seen when comparing patients of the BPH group with the control patients (p=0.019). Resistive index values of the TZ in diabetic patients showed significantly higher values (p<0.001) than the BPH and control groups. CONCLUSIONS/INTERPRETATION: The significantly lower CPI and higher resistive index values of the TZ in diabetic patients compared with patients with non diabetic BPH and healthy subjects indicate considerable vascular damage in the TZ of these patients. Diabetic vascular damage may cause hypoxia and may contribute to the pathogenesis of BPH. PMID- 15756539 TI - Suppression of Pdx-1 perturbs proinsulin processing, insulin secretion and GLP-1 signalling in INS-1 cells. AB - AIMS/HYPOTHESIS: Mutations in genes encoding HNF-4alpha, HNF-1alpha and IPF-1/Pdx 1 are associated with, respectively, MODY subtypes-1, -3 and -4. Impaired glucose stimulated insulin secretion is the common primary defect of these monogenic forms of diabetes. A regulatory circuit between these three transcription factors has also been suggested. We aimed to explore how Pdx-1 regulates beta cell function and gene expression patterns. METHODS: We studied two previously established INS-1 stable cell lines permitting inducible expression of, respectively, Pdx-1 and its dominant-negative mutant. We used HPLC for insulin processing, adenovirally encoded aequorin for cytosolic [Ca2+], and transient transfection of human growth hormone or patch-clamp capacitance recordings to monitor exocytosis. RESULTS: Induction of DN-Pdx-1 resulted in defective glucose stimulated and K+-depolarisation-induced insulin secretion in INS-1 cells, while overexpression of Pdx-1 had no effect. We found that DN-Pdx-1 caused down regulation of fibroblast growth factor receptor 1 (FGFR1), and consequently prohormone convertases (PC-1/3 and -2). As a result, DN-Pdx-1 severely impaired proinsulin processing. In addition, induction of Pdx-1 suppressed the expression of glucagon-like peptide 1 receptor (GLP-1R), which resulted in marked reduction of both basal and GLP-1 agonist exendin-4-stimulated cellular cAMP levels. Induction of DN-Pdx-1 did not affect glucokinase activity, glycolysis, mitochondrial metabolism or ATP generation. The K+-induced cytosolic [Ca2+] rise and Ca2+-evoked exocytosis (membrane capacitance) were not abrogated. CONCLUSIONS/INTERPRETATION: The severely impaired proinsulin processing combined with decreased GLP-1R expression and cellular cAMP content, rather than metabolic defects or altered exocytosis, may contribute to the beta cell dysfunction induced by Pdx-1 deficiency. PMID- 15756543 TI - Cervical ankylosis following Grisel's syndrome in a 14-year-old boy with infectious mononucleosis. AB - Non-traumatic atlanto-axial subluxation (Grisel's syndrome) is an uncommon complication of neck space infection or otolaryngologic procedures. It most frequently affects children, although it may occur in adults. We present a 14 year-old boy with Grisel's syndrome as a complication of infectious mononucleosis proceeding to cervical ankylosis. PMID- 15756542 TI - Seeding of osteosarcoma in the biopsy tract of a patient with multifocal osteosarcoma. AB - We report a case of multifocal osteosarcoma in a 7-year-old boy who developed iatrogenic seeding of tumor along the biopsy tract. The results of the plain radiograph, CT, and histopathological correlation are presented. PMID- 15756544 TI - Spurious leptomeningeal enhancement on immediate post-operative MRI for paediatric brain tumours. AB - Immediate post-operative MRI has been recommended as an accurate and robust method to assess residual brain tumour. Early enhancement at the resection margin and in the dura is well recognized, but we describe two cases of enhancement in the basal cisterns on immediate post-operative MRI that resolved on follow-up. The underlying cause of the enhancement remains to be elucidated, but it should be recognized that leptomeningeal enhancement may occur after surgery and that this does not necessarily imply tumour spread. PMID- 15756545 TI - Evolution of transcriptional enhancers in the immunoglobulin heavy-chain gene: functional characteristics of the zebrafish Emu3' enhancer. AB - Past studies of the channel catfish immunoglobulin heavy-chain (IgH) locus indicates that it lacks an Emu enhancer in the J(H)-Cmu1 intron but does have an enhancer, termed Emu3', in the mu-delta intergenic region. The positioning of the catfish enhancer downstream of the mu-chain exons is predicted to be unfavorable for antibody-affinity maturation in catfish, and would also have been an impediment to the evolution of class switch recombination, had it existed in early tetrapods. To determine if this downstream enhancer is a general feature of teleost fish, we have identified the location of the transcriptional enhancer in the zebrafish IgH locus. We find that zebrafish, like catfish, only have an Emu3' like enhancer that has cross-species activity, but which is B-cell-specific in its activity. A 300-bp region of the zebrafish enhancer shares sequence homology with the core of the catfish Emu3', although there has been loss and gain of specific octamer enhancer motifs. Mutagenesis studies demonstrate that the zebrafish IgH enhancer depends on a pair of E-box motifs that are found in the enhancer core. Similarly spaced E-box motifs appear to exist in the Emu3' enhancer regions of other teleost fish, suggesting this is a common feature among fish IgH enhancers. We discuss how this distal positioning of the enhancer may influence affinity maturation in extant teleosts as well as the evolution of this process in the early tetrapods. PMID- 15756546 TI - Recombination and the origin of sequence diversity in the DRB MHC class II locus in chamois (Rupicapra spp.). AB - We examined the evolutionary processes contributing to genetic diversity at the major histocompatibility complex (MHC) class II DRB locus in chamois (Rupicapra spp., subfamily Caprinae). We characterised the pattern of intragenic recombination (or homologous gene conversion) and quantified the amount of recombination in the genealogical history of the two chamois species, Pyrenean chamois (Rupicapra pyrenaica) and Alpine chamois (Rupicapra rupicapra). We found evidence for intragenic recombination, and the estimated amount of population recombination suggests that recombination has been a significant process in generating DRB allelic diversity in the genealogical history of the genus Rupicapra. Moreover, positive selection appears to act on the same peptide binding residues in both analysed chamois species, but not in identical intensity. Recombination coupled with positive selection drives the rapid evolution at the peptide-binding sites in the MHC class II DRB gene. Many chamois MHC class II DRB alleles are thus much younger than previously assumed. PMID- 15756549 TI - Characterization and expression analysis of CD3varepsilon and CD3gamma/delta in fugu, Takifugu rubripes. AB - CD3 is an essential component of the CD3-TCR complex. In this report, we describe the cloning, characterization, and expression analysis of the CD3varepsilon and CD3gamma/delta chain genes from fugu, Takifugu rubripes. Two distinct CD3varepsilon homologue cDNAs, designated as CD3varepsilon-1 and CD3varepsilon-2, and a CD3gamma/delta homologue cDNA were isolated from the fugu thymus. The deduced amino acid sequences of these cDNAs exhibit conserved essential CD3 chain motifs and overall structures. RT-PCR analysis demonstrated that the CD3varepsilon and CD3gamma/delta genes were expressed in lymphoid organs (e.g. thymus, head kidney, trunk kidney and spleen), mucosal tissues (gill, skin, and intestine), and peripheral blood leucocytes (PBL). The CD3 and TCRalpha genes were expressed only in the surface IgM- population, which were separated from PBL using an anti-fugu IgM monoclonal antibody. In addition, in situ hybridization confirmed that CD3-expressing cells were distributed randomly in the head kidney, trunk kidney, and spleen, but in the thymus were restricted to the lymphoid outer zone and epithelioid inner zone only. Collectively, these results suggest that CD3 molecules are useful markers for the identification of T cells in teleost fish. The present study thus provides a critical step in identifying T cells in this model organism. PMID- 15756550 TI - [Regression and therapy-resistance of primary liver tumors and liver metastases after regional chemotherapy and local tumor ablation]. AB - High dosage regional chemotherapy, chemoembolization and other methods of regional treatment are commonly used to treat unresectable primary liver malignancies and liver metastases. In liver malignancies of childhood neoadjuvant chemotherapy is successfully combined with surgical treatment. Chemotherapy and local tumor ablation lead to characteristic histomorphologic changes: Complete destruction of the tumor tissue and its vascular bed is followed by encapsulated necroses. After selective eradication of the tumor cells under preservation of the fibrovasular bed the tumor is replaced by hypocellular edematous and fibrotic tissue. If completely damaged tumor tissue is absorbed quickly, the tumor area is replaced by regenerating liver tissue. Obliterating fibrohyalinosis of tumor vessels, and perivascular edema or necrosis indicate tissue damage along the vascular bed. Degenerative pleomorphism of tumor cells, steatosis, hydropic swelling and Malloryhyalin in HCC can represent cytologic findings of cytotoxic cellular damage. Macroscopic type of HCC influences significantly the response to treatment. Multinodular HCC often contain viable tumor nodules close to destroyed nodules after treatment. Encapsulated uninodular tumors undergo complete necrosis much easier. Large size and a tumor capsule limitate the effect of percutaneous injection of ethanol into HCC. In carcinomas with an infiltrating border, especially in metastases of adenocarcinomas and hepatic cholangiocarcinoma cytostatic treatment damages the tumor tissue mainly in the periphery. Nevertheless the infiltrating rim, portal veins, lymphatic spaces and bile ducts as well as the angle between liver capsule, tumor nodule and bordering parenchyma are the main refugees of viable tumor tissue even after high dosage regional chemotherapy. This local resistance is caused by special local conditions of vascularization and perfusion. These residues are the source of local tumor progression and distant metastases. Besides intrinsic cellular mechanisms architectural, and microenvironmental factors relevantly limitate the effect of intensive locoregional therapy. PMID- 15756551 TI - Congenital anomalies of the coronary arteries: imaging with contrast-enhanced, multidetector computed tomography. AB - The objective of this study is to evaluate multidetector CT (MDCT) in detecting and characterizing anomalous coronary arteries. Forty-four patients with anomalies of the coronaries were selected from a total of 1758 individuals examined with ECG-gated 4- and 16-row MDCT including thinMIP, MPR and VRT post processing. Twenty-eight patients showed origin and course anomalies of the central coronary segments, and in this subgroup 13 were judged as "malignant" because of interarterial courses between the aortic root and the pulmonary trunk, either of the right coronary artery (n=11) or the left coronary artery (n=2). Twelve non-hemodynamic anomalies were found, affecting the coronary origins only (n=10) or the peripheral vessels courses (n=2). Four arteriovenous fistulas were present, all of them with complex arterial feeders. Regardless of vessel anatomy, coronary opacification was always possible by means of the systemic contrast agent, and the aberrant coronary arteries were visualized synoptically in direct relation to the great mediastinal vessels. In contrast to MDCT, selective cannulation and final diagnosis was possible in only 11 of the 20 catheter angiograms performed (accuracy of 55.0%). In conclusion, its non-invasiveness and precise visualization makes MDCT the standard of reference for evaluating anomalous coronary arteries. PMID- 15756552 TI - Variation of the size of pulmonary venous ostia during the cardiac cycle: optimal reconstruction window at ECG-gated multi-detector row CT. AB - The aim of this study was to investigate the variation of the size of pulmonary vein ostia during cardiac cycle using ECG-gated multi-detector row CT (MDCT). Nineteen patients were included in this study. Transaxial images at the level of right inferior pulmonary vein (RIPV) were reconstructed in increments of 5%. The ostial diameter of RIPV was measured, the reconstruction windows showing maximal and minimal diameters were selected. The ostial areas of four pulmonary veins were measured at axial image sets of two selected reconstruction windows. The measurement of RIPV revealed that the maximal diameter (1.50+/-0.32 cm) was generally 35% and the minimal diameter (1.28+/-0.28 cm) was usually at 85%. The measurement of ostial areas showed that the ostia enlarged at the end of ventricular systole when compared with those at the end of ventricular diastole, by the factors of 1.44+/-0.55 for the right superior, 1.25+/-0.23 for the right inferior, 1.45+/-0.81 for the left superior, and 1.31+/-0.26 for the left inferior pulmonary vein (P<0.05). The size of the pulmonary vein ostia is variable during the cardiac cycle and the measurement of the pulmonary veins should always be in the same phase of the cardiac cycle during the follow-up of patients. PMID- 15756553 TI - Percutaneous radiofrequency ablation of renal cell carcinoma. AB - Conventional curative therapy for renal cell carcinoma has been open nephrectomy. However, several less invasive and/or nephron-sparing procedures have been developed as alternatives in selected patients. The newest of these therapeutic modalities involves percutaneous image-guided ablation with straight or expandable needle applicators that deposit energy. Radiofrequency ablation is the modality for which there is the largest reported experience with percutaneous application, and involves the use of electrical current to generate frictional heating of tissue. Animal studies confirm the ability of radiofrequency ablation to cause regions of necrosis within normal kidney and in VX2 tumors. Clinically, radiofrequency ablation of small renal cell carcinoma is increasingly being performed in selected patients who are not ideal surgical candidates. Results are excellent for small exophytic tumors, but successful treatment is less likely as tumor size increases or the location becomes more central. Complete treatment of most tumors requires one or more overlapping ablations with the needle electrodes positioned so as to cause necrosis in the entire volume of tumor. The number of overlapping ablations and the position of the needle electrodes vary based on tumor size and geometry. For very large tumors, pre-ablation catheter embolization may enhance the results of ablation by decreasing blood flow and perfusion mediated cooling. Following ablation, imaging with CT or MR is performed to assess the result and to diagnose any residual tumor so that it can be treated. Because the tumor remains in situ, imaging follow-up continues indefinitely. The complication rate of radiofrequency ablation is favorable when compared to surgical techniques. Long-term survival data are not yet available. PMID- 15756554 TI - Sonographic imaging of the paediatric female pelvis. AB - Several conditions necessitate a pelvic ultrasound in their management. In this paper, we will first review the embryology of the female genital system and the normal morphology of the uterus and ovaries from birth to puberty. Thereafter, this paper aims to categorise pelvic pathologies based on their clinical presentations. We will consider successively ambiguous genitalia, precocious puberty, delayed puberty and amenorrhea, gynaecological masses and, finally, pelvic pain of gynaecological origin. Ultrasound is the key screening tool and often the only examination indicated. The appropriateness of other modalities, such as computed tomography and magnetic resonance imaging, will be discussed. The aim of this article is to help radiologists form a sensible imaging plan when presented with a diagnostic dilemma involving the pelvis in female children. PMID- 15756555 TI - Radiofrequency thermoablation of primary non-spinal osteoid osteoma: optimization of the procedure. AB - Osteoid osteoma is a small benign tumor that requires treatment due to the intense pain it causes. Surgical therapy has been the ablative technique of choice after a failure of medical therapy. Recently, numerous less invasive, alternative procedures have been proposed: drill trepanation with or without ethanol injections, cryoablation, and thermoablation with laser or radiofrequency. The aim of this review is to retrospectively assess the effect of radiofrequency (RF) thermoablation in the treatment of primary non-spinal osteoid osteoma. From June 2001 to July 2003, we treated 106 patients affected by osteoid osteoma with RF thermoablation. Five patients with spinal osteoid osteoma and four with a previously treated osteoma were excluded from the study. In this paper, we assess the results obtained in a selected group of 97 primary non spinal osteoid osteoma. The lesions were predominantly in the metaphysics of the femur. Central nidus calcifications were frequent and there was no prevalence for which side they occurred. Primary success was achieved in 82 patients (85%), while we obtained secondary success in 15 patients (15%). In two patients (2%), pain persisted between the two treatments and failed to be resolved, even after the second treatment; therefore, surgical excision was performed and complete resolution was obtained. No complications were reported. In conclusion, our results confirm that the treatment of choice for non-spinal osteoid osteoma is RF thermoablation, offering several advantages over ablative techniques. PMID- 15756556 TI - [Application of EMLA creme before upper lid blepharoplasty]. AB - BACKGROUND: The injection of local anesthesia before upper lid blepharoplasty is perceived as unpleasant and painful by some patients. METHODS: In an intraindividual randomized prospective study, 14 patients undergoing upper lid blepharoplasty on both sides were examined. Before the operation the EMLA creme was put on one side. The pain sensibility was checked before and after using EMLA creme and during the infiltration of anesthesia and the operation. RESULTS: The sensitivity before putting on EMLA creme was equal in all patients on both sides. After application there was a distinct difference between the sides. During the infiltration of anesthesia 12 patients reported low and 2 patients medium pain on the treated side, while on the non-treated side 5 patients reported low, 4 patients medium, and 5 patients severe pain. CONCLUSIONS: Applying EMLA creme before an operation reduces the pain sensitivity during infiltration of anesthesia and during the operation. PMID- 15756557 TI - [Diagnostic value of nerve fibre layer photography in glaucoma]. AB - PURPOSE: Primary open-angle glaucoma is characterized by a loss of retinal nerve fibres. The aim of our present study was to evaluate the diagnostic value of retinal nerve fibre layer (RNFL) photography as a routine examination for the early diagnosis of glaucomatous damage. METHODS: Between 1991 and 2000 we monitored 96 patients with ocular hypertension in a prospective study. RNFL photography was performed in 91 subjects. These photos were evaluated by two independent observers. RESULTS: The mean image quality of the photos was mediocre. The correlation coefficient for the intraindividual reproducibility (image quality) was 0.713-0.863 (Kendall's coefficient). For the interindividual reproducibility the correlation coefficient was 0.541-0.581 (Somers' coefficient). Regarding the evaluation "RNFL defect yes/no", the correlations were better but there was a discrepancy between the observers. CONCLUSION: Quality, reproducibility and prognostic value of RNFL photography were disappointing. Although RNFL photography may be convincing and informative in several situations, it is not efficient as a routine technique for the early diagnosis of RNFL damage in glaucoma patients. PMID- 15756558 TI - The role of laparoscopy in the diagnosis of intestinal vascular anomalies affecting two small infants. AB - Two infants affected by intestinal vascular anomalies causing lower gastrointestinal hemorrhage are reported. Upper and lower gastrointestinal tract endoscopy and radionuclide scan were negative. The authors found laparoscopic exploration very useful in detecting the cause and the site of bleeding. Because the usual investigations for gastrointestinal bleeding have been proven to be time-consuming and often unreliable in small infants, the authors suggest considering laparoscopic exploration as a first-line diagnostic tool in this subset of patients. A new diagnostic algorithm for gastrointestinal bleeding in infants is proposed. PMID- 15756560 TI - Acute appendicitis presenting as thigh abscess in a child: a case report. AB - A case of retrocecal appendicitis is described in a 6-year-old male child who presented with thigh abscess. The presence of a positive psoas stretch test, feculent discharge, an enteric growth on bacteriological examination, and intraabdominal fluid collection on abdominal ultrasound provided clues to the presence of an intraabdominal source of sepsis. Laparotomy revealed a perforated retrocecal appendix with surrounding collection communicating into the thigh. Appendectomy with drainage of retroperitoneal and thigh collections under adequate antibiotic coverage resulted in a satisfactory recovery. We describe our experience with the present case and discuss the pertinent literature. PMID- 15756559 TI - Is the testis intraperitoneal? AB - The relationship of the testis to the peritoneal cavity, and hence its position as an intraperitoneal or extraperitoneal organ, remains controversial. Adult anatomy texts favour an extraperitoneal position during and after testicular descent, whereas journal articles favour an intraperitoneal position. Interestingly, there is no similar debate around the position of the ovary despite the common origin of each as indifferent gonads. Through direct observation and the literature review, we aimed to determine whether the testis should be considered an intraperitoneal or an extraperitoneal organ. The anatomical and embryological literature relevant to human and animal models was reviewed. Direct dissections were made in rats (n=8) during foetal development, postnatally, and in mature animals, allowing comparison of foetus with adult and male with female. The position of the human testis was also recorded in various pathological states. Direct dissection in rats reveals an intraperitoneal testis on a mesorchium during both foetal and postnatal life. Intraperitoneal testes are demonstrated in humans in cases of gastroschisis (where the testis may protrude through the periumbilical defect with the bowel), testicular torsion (where the testis is mobile within the peritoneum), and bell clapper testis (where the testes are identifiable as intraperitoneal). We conclude that the foetal testis is an intraperitoneal organ. In the adult rat the testis remains intraperitoneal. The postnatal human testis is intraperitoneal. The adult human testis is intraperitoneal but may appear extraperitoneal. The apparent discrepancy between the adult testis being intraperitoneal or extraperitoneal is likely to result from differences in the relative size of the tunica vaginalis between infant boys and elderly men. PMID- 15756562 TI - Effect of subcutaneous insulin on intestinal adaptation in a rat model of short bowel syndrome. AB - Insulin has been shown to influence intestinal structure and absorptive function. The purpose of the present study was to evaluate the effects of parenteral insulin on structural intestinal adaptation, cell proliferation, and apoptosis in a rat model of short bowel syndrome (SBS). Male Sprague-Dawley rats were divided into three experimental groups: sham rats underwent bowel transection and reanastomosis, SBS rats underwent a 75% small bowel resection, and SBS-INS rats underwent a 75% small bowel resection and were treated with insulin given subcutaneously at a dose of 1 U/kg, twice daily, from day 3 through day 14. Parameters of intestinal adaptation, enterocyte proliferation, and enterocyte apoptosis were determined on day 15 following operation. SBS rats demonstrated a significant increase in jejunal and ileal bowel and mucosal weight, villus height and crypt depth, and cell proliferation index compared with the sham group. SBS INS animals demonstrated higher jejunal and ileal bowel and mucosal weights, jejunal and ileal mucosal DNA and protein, and jejunal and ileal crypt depth compared with SBS animals. SBS-INS rats also had a greater cell proliferation index in both jejunum and ileum and a trend toward a decrease in enterocyte apoptotic index in jejunum and ileum compared with the SBS untreated group. In conclusion, parenteral insulin stimulates structural intestinal adaptation in a rat model of SBS. Increased cell proliferation is the main mechanism responsible for increased cell mass. PMID- 15756563 TI - Expression of vasoactive mediators during mechanical ventilation in nitrofen induced diaphragmatic hernia in rats. AB - The high mortality in patients with congenital diaphragmatic hernia (CDH) has been attributed to pulmonary hypoplasia and persistent pulmonary hypertension (PPH). Endothelin-1 (ET-1), nitric oxide (NO), and calcitonin gene-related peptide (CGRP) have been reported to be important vasoactive mediators in the perinatal pulmonary circulation. The exact mechanism by which these vasoactive mediators interact to regulate the perinatal pulmonary vascular tone in CDH with PPH is not fully understood. We hypothesized that the altered pulmonary vascular reactivity in CDH is due to imbalance in vasoactive mediators. This study was designed to investigate mRNA expression of ET-1, eNOS, and CGRP in CDH lung in the perinatal period. A CDH model was induced in pregnant rats following administration of nitrofen. In control animals, the same dose of olive oil was given without nitrofen. Cesarean section was performed on day 21 of gestation. The newborn rats were intubated and ventilated, and ventilation was continued for 1-6 h. Left lungs were collected from both groups at 0, 1, and 6 h after ventilation (n=8 in each group). Reverse transcriptase-polymerase chain reaction on lung tissue was performed to evaluate the relative level of ET-1, eNOS, and CGRP mRNA expression. The results showed a significant increase in ET-1 mRNA in CDH lung at 1 and 6 h after ventilation compared with controls. In CDH lung, eNOS mRNA and CGRP mRNA levels were significantly increased at 1 h but were similar to control values at 6 h after ventilation. The increased expression of vasoconstrictor ET-1 mRNA and vasodilators eNOS mRNA and CGRP mRNA in the CDH lung at 1 h after ventilation suggests that pulmonary vascular tone is rapidly changing after birth. An imbalance in the production of vasoconstrictors and vasodilators by the CDH lung may contribute to high pulmonary vascular resistance. PMID- 15756564 TI - The effect of vagal neural crest ablation on the chick embryo cloaca. AB - The cloaca, the caudal limit of the avian gastrointestinal tract, acts as a collecting chamber into which the gastrointestinal, urinary, and genital tracts discharge. It is intrinsically innervated by the enteric nervous system, which is derived from neural crest emigres that migrate from the vagal and sacral regions of the neural tube. Abnormal cloacal development can cause a number of anorectal anomalies, including persistent cloaca. Ablation of the vagal neural crest has previously been shown to result in an aganglionic hindgut to the extent of the colorectum. The aim of our study was to investigate the effect of vagal neural crest ablation on the cloaca, the limit of the hindgut in the developing chick embryo. Chick embryos were incubated until the 10-12 somite stage. The vagal neural tube corresponding to the level of somites 3-6 was then ablated, and eggs were incubated until harvested on embryonic day 11 (E11). Whole chick embryos were fixed, embedded in paraffin, and sectioned. Immunohistochemistry was then carried out using the HNK-1 monoclonal antibody to label neural crest cells, and results were assessed by light microscopy. Vagal neural crest ablation resulted in a dramatic decrease in the number of neural crest cells colonizing the chick embryo cloaca compared with control embryos. Ablated embryos contained only a small number of HNK-1-positive neural crest cells, which were scattered within the myenteric plexus in a disorganised pattern. Hypoganglionosis was also evident in other regions of the hindgut in ablated embryos. Ablation of the vagal neural crest results in a hypoganglionic cloaca in addition to hypoganglionosis of the hindgut. These results suggest that the cloaca is largely innervated by vagal neural crest emigres. Further studies involving quail-chick chimeras to investigate the exact contribution provided by both vagal and sacral neural crest cells to the cloaca should increase our understanding of the pathophysiology of conditions like persistent cloaca. PMID- 15756566 TI - Critical presentation of pleuropulmonary blastoma. AB - We report two cases of critical presentation of a quite rare lung neoplasm of childhood. Presentation findings were at the extremes of the clinical pattern of this polymorphous neoplasm, ranging from an enormous solid mass causing airway compression and dislocation to an apparently benign cystic lesion discovered because of a tension pneumothorax. Both children were discharged from the pediatric intensive care unit and underwent appropriate surgical removal and oncological management. Congenital lung cysts, even asymptomatic, should not be underestimated and need elective surgical excision and histologic examination. PMID- 15756565 TI - Bacterial translocation in acute rejection after small bowel transplantation in rats. AB - Acute rejection after small bowel transplantation (SBTx) may facilitate bacterial translocation (BT) and subsequent changes in the liver, spleen, and lungs. This study investigated whether BT occurs after acute rejection and whether this is followed by changes in the structure of the intestine and the phagocytic organs interposed between the gut and the general circulation. Orthotopic SBTx was performed in allogeneic (ALLO) rat-strain combinations (BN-Wistar, n=5). For comparison we used syngeneic SBTx (SYN) (BN-BN, n=6) controls. Animals were sacrificed on postoperative day 7. Mesenteric lymph nodes and portal and caval blood were cultured for aerobes and anaerobes. Escherichia coli beta galactosidase DNA was assessed by polymerase chain reaction in the blood samples. Intestine, liver, spleen, and lung protein and DNA contents were measured. Histologic changes were graded according to standard criteria of acute rejection. For comparisons we used chi(2) and nonparametric Mann-Whitney test with a threshold of significance of p<0.05. ALLO rats lost more weight after SBTx than SYN rats (-13.02+/-4.39% vs. -8.04+/-5.08% of preoperative weight), although the difference was not significant (ns). A variable degree of graft rejection was histologically demonstrated in all ALLO rats, and DNA/protein content in the graft was significantly higher in this group (0.245+/-0.85 vs. 0.134+/-0.21, p<0.05). Gram-negative enteric bacteria were found in 4/5 ALLO and 4/6 SYN rats (ns), and aerobic Gram-positive bacteria in 2/5 and 3/6 (ns), respectively. Anaerobic growth occurred in mesenteric lymph nodes in one ALLO rat and in the bloodstream in another one. E. coli DNA was isolated in none of the ALLO but in two SYN rats (ns). BT was frequent after SBTx in both syngeneic and allogeneic strain combinations. Contrary to our expectations, BT after SBTx was not higher in ALLO group rats. However, anaerobic germs were isolated only in this group. PMID- 15756567 TI - Morphology and glycoconjugate histochemistry of the eccrine glands in the snout skin of the North American raccoon (Procyon lotor). AB - The eccrine nasolabial glands were found in the hypodermis of the nasal plane in the North American raccoon (Procyon lotor). In addition to light and electron microscopic observations, the distribution and selectivity of complex glycoconjugates in the eccrine tubular glands of the raccoon snout skin were studied using various histochemical methods, particularly lectin staining. The secretory epithelium and the luminal secretions exhibited high amounts of glycoconjugates with various saccharide residues (alpha-D: -mannose, alpha-L: fucose, beta-D: -galactose, beta-N-acetyl-D: -glucosamine, sialic acid). The excretory duct cells also showed positive reactions with most of the histochemical methods applied. The results are discussed with regard to possible functions of the glandular secretions. The complex glycoconjugates that are produced by the eccrine nasolabial glands may be related to moistening of the skin surface as well as protecting the epidermis against physical damage or microbial contamination. This is the first report on the glands in the snout skin of carnivores. PMID- 15756569 TI - Deep neck infection due to Lactococcus lactis cremoris: a case report. AB - This report describes the first case of deep neck infection resulting from Lactococcus lactis subsp. cremoris. The case is associated with the consumption of unpasteurized milk and occurred in a patient with a buccal mucosa tumor. Anti infective therapy with ceftriaxone and metronidazole resulted in complete resolution. PMID- 15756568 TI - Cochlear implantation and management of chronic suppurative otitis media: single stage procedure? AB - In a series of 360 patients who underwent cochlear implantation at our center, four patients (five procedures) had cochlear implantation with obliteration of the mastoid cavity and management of cholesteatoma as a single-staged procedure. Three patients were bilaterally deaf secondary to CSOM and had bilateral mastoid cavities, and in one patient congenital cholesteatoma was identified during cochlear implantation. A mastoidectomy or revision mastoidectomy with obliteration of the mastoid cavity and cochlear implantation was performed as a single stage procedure. Cholesteatoma reoccurred in one patient 9 years after cochlear implantation. Surgical procedures, complications, follow-up and outcomes are discussed. PMID- 15756572 TI - Dynamic contour tonometry versus Goldmann applanation tonometry: a comparative study. AB - BACKGROUND: Various sources of error, including central corneal thickness (CCT) and structural corneal rigidity, have been proposed for Goldmann applanation tonometry (GAT). The Pascal dynamic contour tonometer (DCT) is a novel device designed for intraocular pressure (IOP) measurements assumed to be largely independent of CCT and corneal curvature. We compared DCT with GAT in eyes with normal corneas of various thickness. METHODS: We prospectively measured IOP using DCT and GAT in random order in 100 eyes of 100 subjects (M:F=46:54; mean age 42+/ 19, range 23-88 years). RESULTS: Mean DCT values were about 1 mm Hg higher than GAT readings (16+/-3 vs 15+/-3 mmHg, p=0.001). Bland-Altman analysis of individual pairs of DCT and GAT measurements revealed a bias of -1.0 mmHg [95% confidence interval (CI): +/-1.2]. Neither GAT nor DCT showed a significant correlation with CCT (533+/-48, range 399-641 microm). CONCLUSIONS: In eyes with normal corneas, DCT allows suitable and reliable IOP measurements which are in good concordance with GAT. Comparison of DCT with intracameral manometry is desirable in the future. PMID- 15756571 TI - Comparison of prednisolone 1%, rimexolone 1% and ketorolac tromethamine 0.5% after cataract extraction: a prospective, randomized, double-masked study. AB - PURPOSE: To compare the efficacy, safety and patient comfort of two topical steroids (prednisolone 1% and rimexolone 1%) and a topical non-steroidal anti inflammatory agent (ketorolac tromethamine 0.5%) after extracapsular cataract extraction. METHODS: Forty-five patients were enrolled in this prospective, randomized, double-blind study. They were assigned to receive topical treatment with either prednisolone, rimexolone or ketorolac tromethamine ophthalmic solution after phacoemulsification for cataract extraction. On postoperative days 1, 3, 5, 14 and 28 best-corrected visual acuity, intraocular pressure (IOP), slit lamp examination of the anterior segment and report of the patients' comfort were assessed and compared by Friedman rank time analysis. RESULTS: Regarding the primary outcome efficacy of inflammation control the assessment of cells did not differ (p=0.165), while flare in the anterior chamber was lowest (p=0.008) in the non-steroidal anti-inflammatory drug (NSAID) group. Surface inflammation was lowest with prednisolone (p=0.002). Regarding safety, visual acuity did not differ among the groups. In the prednisolone group one patient, however, responded to steroid treatment with elevated IOP and had to be excluded. In the remaining patients IOP was even lower in the two steroidal treatment groups than with ketorolac (p=0.030). One patient receiving ketorolac had to be excluded because a corneal erosion developed. Patient comfort was highest with prednisolone (p=0.041). CONCLUSIONS: Ketorolac tromethamine provides good control of intraocular inflammation after cataract extraction without the risk of a steroidal IOP increase, which was also not observed under rimexolone therapy. The best surface inflammation control and patient comfort was observed with prednisolone, which remains a good choice. PMID- 15756573 TI - Cavernous sinus thrombosis elicited by a central retinal vein venous stasis retinopathy. AB - BACKGROUND: Central retinal vein occlusion is a relatively common retinal disorder in the elderly, and those with cardiovascular or thrombophilic risk factors are at increased risk. Although still unsatisfying, some treatments for the acute and chronic phases have been established based on randomized studies. However, for rare conditions mimicking central retinal vein occlusion, treatment of the acute phase should be targeted at etiology. A rare condition mimicking central retinal vein occlusion in a 70-year-old man is presented and discussed. METHODS: A 70-year-old man was admitted to the hospital for isovolemic hemodilution related to a central retinal vein occlusion of the elderly, after a sudden decrease in visual acuity to 0.1. RESULTS: Clinical and laboratory work-up demonstrated a venous stasis retinopathy, related to an atypical cavernous sinus thrombosis of undetermined origin. The patient had experienced in the past 6 months intermittent diplopia and an irreducible conjunctival hyperemia. Hemodilution was dismissed. Soon after initiation of anticoagulation therapy, the patient's clinical signs and symptoms improved. Final visual acuity was 0.8. CONCLUSION: Venous stasis retinopathy secondary to cavernous sinus thrombosis is rare. However, careful clinical examination and extensive laboratory work-up is needed to exclude central retinal vein occlusion not associated with common vascular pathologies of the elderly. PMID- 15756574 TI - Scanning laser polarimetry with variable corneal compensation and detection of glaucomatous optic neuropathy. AB - BACKGROUND: The aim of this study was to evaluate the ability of scanning laser polarimetry (SLP) parameters provided by commercially available GDx with variable corneal compensator (VCC) to discriminate between healthy and glaucomatous eyes. METHODS: Sixty-five healthy and 59 glaucomatous age-matched patients underwent a complete ophthalmological evaluation, an achromatic automated perimetry (AAP), and SLP with GDx-VCC. One randomly selected eye from each subject was considered. All glaucomatous eyes had reproducible visual field defects. Mean values (+/- SD) of all SLP-VCC parameters measured in the two groups were compared. Area under receiver operating characteristics (AUROC) curve and sensitivities at predetermined specificities of >or=80% and >or=95% for each single parameter were calculated. Moreover, the nerve fiber indicator (NFI) diagnostic accuracy was evaluated calculating positive, negative, and interval likelihood ratios (LRs) at different cutoff values. RESULTS: All SLP parameters were significantly different between the two groups (p<0.001). The NFI showed the best AUROC curve (0.938, SE 0.02) whereas temporal, superior, nasal, inferior, temporal (TSNIT) average was second best (0.897, SE 0.03), and normalized superior area was third (0.879, SE 0.04). At fixed specificity >or=95%, sensitivities ranged from 22% to 79.7% whereas for values >or=80%, sensitivities were in the 44.1-89.8% range. At a cutoff NFI value of 30, positive LR was 17.6 (95% CI: 5.8-53.6) and negative LR was 0.19 (95% CI: 0.11-0.33). Interval LRs for NFI showed that values 40 were associated with large effects on posttest probability. CONCLUSIONS: SLP VCC allows good discrimination between healthy and glaucomatous eyes. New software-provided parameters NFI, TSNIT average, and normalized superior and inferior areas appear to be reliable in the evaluation of glaucomatous disease. In particular, after evaluation on interval LRs, the NFI showed a high diagnostic accuracy for values 40. PMID- 15756575 TI - The course of epiphora after failure of silicone intubation for congenital nasolacrimal duct obstruction. AB - BACKGROUND: One of the indications for dacryocystorhinostomy (DCR) in children with congenital nasolacrimal duct obstruction (CNLDO) is failure of silicone intubation. We evaluated the course of epiphora after failure of silicone intubation for CNLDO when DCR was not performed. METHODS: In a comparative cohort study carried out at a tertiary referral center, ten eyes of seven consecutive children who had failure of silicone intubation manifested as persistent epiphora over 2 months and whose parents refused DCR were followed up for an average of 50.4 months (range 33-70 months). Three lacrimal drainage systems of three other children who had failure of silicone intubation underwent uneventful DCR. RESULTS: In eight (80%) of the ten consecutive eyes with congenital nasolacrimal duct obstruction (six of the seven children, 86%), there was spontaneous complete resolution of the epiphora and normal dye disappearance test (DDT) at the end of the follow-up period. One child with Down's syndrome, allergic rhinitis, asthma and multiple site obstructions had improvement of symptoms but abnormal DDT. The epiphora in all three children who underwent DCR had disappeared by 6 months after surgery when the silicone tube was removed. No complications were noted during the follow-up. CONCLUSIONS: Epiphora can spontaneously resolve after failure of silicone intubation in CNLDO, and DCR should no longer considered be compulsory in such cases unless complications evolve. PMID- 15756577 TI - Diurnal IOP fluctuation: not an independent risk factor for glaucomatous visual field loss in high-risk ocular hypertension. AB - PURPOSE: To establish whether intraocular pressure (IOP) fluctuations contribute to the risk of developing glaucoma in patients with high-risk ocular hypertension. METHODS: Ninety patients included in the Malmo Ocular Hypertension Study were examined every 3 months with office-hours diurnal tension curves and computerised perimetry. Patients were followed up prospectively for 10 years or until glaucomatous visual field loss could be demonstrated. Poststudy data were included in the analyses, extending maximum follow-up to 17 years. RESULTS: After 17 years, 37 patients had developed glaucomatous visual field defects. When applying univariate Cox regression analyses, mean IOP of all measurements during the prospective part of the study was a significant risk factor for developing glaucoma (95% confidence interval [CI] 1.08-1.39), while IOP fluctuations were almost significant (95% CI 0.98-1.93). When separating effects of mean IOP level and mean IOP fluctuation using Cox multiple regression analysis, only IOP level came out as significant (95% CI 1.09-1.38), and IOP fluctuations did not contribute to the risk (95% CI 0.80-1.60). IOP fluctuation depended linearly on IOP level (p<0.0001), i.e. IOP fluctuation was larger in eyes with higher IOP levels. CONCLUSION: IOP fluctuations were not an independent risk factor for the incidence of glaucomatous visual field loss in subjects with ocular hypertension. PMID- 15756576 TI - An optimised protocol for the production of autologous serum eyedrops. AB - BACKGROUND: Serum eyedrops have been successfully used in the treatment of severe dry eye, persistent epithelial defects and other severe ocular surface disorders. A number of clinical studies showed a variable efficacy of this approach, but the parameters for the production of this blood product varied significantly. In order to establish an optimised protocol for the production of serum eyedrops, we examined the effect of various clotting times, centrifugation forces, types of diluent and dilutions on the concentration of growth factors, fibronectin, and vitamins in serum and tested the epitheliotrophic capacity of these serum modifications in a cell culture model of human SV-40-immortalised corneal epithelial cells (HCE-T). METHODS: Serum samples were prepared with a clotting time of 20, 60 or 120 min, a centrifugation force of 500 xg or 3,000 xg, and diluted with BSS or isotonic saline. The concentrations of EGF, TGF-beta1, PDGF AB, FGF, HGF, fibronectin, vitamin A and vitamin E in these samples were evaluated with ELISA and HPLC. HCE-T cells were incubated for 24, 48, 72, 96 and 144 h with 100, 50, 25, 12.5, 6.25 and 3.125% serum in diluent, and cell proliferation, migration and differentiation were evaluated by means of a luminescence-based ATP assay, a colony-dispersion assay and scanning electron microscopy. RESULTS: Using a longer clotting time resulted in an increased concentration of all the epitheliotrophic factors examined in serum; the difference was statistically significant for EGF, TGF-beta1 and HGF. Increasing the g force of centrifugation from 500 xg to 3,000 xg resulted in significantly less TGF-beta1, but more EGF and vitamin A. Cell proliferation was better supported by serum prepared with 3,000 xg and diluted with BSS. Serum prepared with a longer clotting time yielded better cell migration and differentiation. CONCLUSION: Clotting time, centrifugation and diluents have a significant impact on the composition and epitheliotrophic effects of serum. A long clotting time (>or=120 min), a sharp centrifugation (3,000 xg for 15 min) and dilution with BSS improve the ability of serum eyedrops to support proliferation, migration and differentiation of corneal epithelial cells. PMID- 15756578 TI - Viagra-associated serous macular detachment. AB - PURPOSE: To describe the OCT findings in two cases of idiopathic serous macular detachments associated with Sildenafil (Viagra) use. METHODS: Two patients were identified with idiopathic serous macular detachments associated with Viagra use. Each patient underwent rigorous historical questioning to rule out known causes of serous macular detachments and to determine contributing factors. A comprehensive ocular examination was performed including dilated biomicroscopic examination of the macula and indirect ophthalmoscopic evaluation of the retinal periphery. Additional evaluation included color fundus photography with FA and ICG, OCT analysis of the macula and B-scan ultrasonography. Systemic workup included MRI of the brain and orbits, CT of the brain, orbits, chest and abdomen, blood chemistries and electrophoresis, lumbar puncture with CSF analysis and chest radiographs. RESULTS: Both patients demonstrated idiopathic serous macular detachment associated with peri-macular vitelliform deposits. Although notable peripapillary atrophy and RPE mottling were present, there was no evidence of heme, exudate, or fibrosis to suggest a CNVM. FA showed no focal areas of leakage. Early and late frames of the ICG demonstrated engorged choroidal vessels without frank leakage. Extensive medical evaluation failed to reveal a systemic etiology. OCT findings demonstrated large serous macular detachments that corresponded to the use or discontinuation of Viagra. Discontinuation of Viagra coincided with complete resolution of the serous macular detachment in one patient and improvement of the serous macular detachment in the other. When one patient resumed Viagra use, he experienced a recurrent serous macular detachment. CONCLUSIONS: We report the OCT findings of two cases of idiopathic serous macular detachment associated with use of the potent vasodilator Sildenafil (Viagra). PMID- 15756579 TI - Relationship of eye burns with calcifications of the cornea? AB - INTRODUCTION: The clinical pattern of corneal calcification has been considered to be a part of chronic eye diseases, such as uveitis, severe glaucoma, keratitis and eye burns. Since, in some cases of eye burns, we have found corneal calcifications to be related to the initial calciferous caustic agents and to the use of phosphate-buffer-containing fluids, we have reviewed our files of clinical eye burns for the incidence and appearance of corneal calcification. METHODS: A total of 176 burnt eyes of 98 patients suffering from severe eye burns were retrospectively reviewed. The following data set was acquired: the type of caustic agent, the time of the burns, the delay and type of immediate treatment and the time and type of first aid, the subsequent medication, the clinical grading and the later results. RESULTS: Calciferous burning agents and corneal calcification are correlated (P<0.01). Initial single rinsing with phosphate does not produce corneal calcification (P=0.134). There is evidence that corneal calcifications are correlated with chronically administered phosphate-buffered eye drops (P<0.005). DISCUSSION: Eye burns followed by calcifications follow two different major patterns: the corrosive substance contained calcium or the continued therapy was applied with phosphate-buffered eye drops. We present case reports of three different types of eye burns and later therapy resulting in corneal calcification. Corneal calcifications are presumably related to longer lasting phosphate application. One suspicious mechanism is the low content of calcium ion stabilizing proteins such as hyaluronate or fetuin in treatments for severe eye burns. The exceeding of the solubility product of Ca(2+) and PO(4) (-) results in the precipitation of calcium phosphates. In cases of chronic corneal disturbance, we recommend the elimination of phosphate-buffered medications to prevent corneal calcification. PMID- 15756580 TI - Intravitreal triamcinolone acetonide in sympathetic ophthalmia. AB - PURPOSE: To report the result of intravitreal triamcinolone acetonide in the treatment of sympathetic ophthalmia. METHODS: A 29-year-old woman who suffered from sympathetic ophthalmia and who was being treated with systemic corticosteroid therapy received an intravitreal injection of 4 mg of triamcinolone acetonide. RESULTS: By the 15th day after injection visual acuity had improved from 20/200 to 20/40 and serous retinal detachment had almost completely resorbed. Systemic corticosteroid therapy was reduced sequentially. By the third month after injection, the patient was in clinical remission. Her visual acuity was 20/20 and no serous detachment was observed. CONCLUSIONS: In this study, short-term improvement in the clinical picture of a patient with sympathetic ophthalmia after intravitreal triamcinolone acetonide injection was described. The results suggest that intravitreal triamcinolone acetonide injection may be an additional tool in the treatment of sympathetic ophthalmia. PMID- 15756581 TI - [Duration of migraine disease correlates with amplitude and habituation of event related potentials]. AB - BACKGROUND: Duration of a migraine disease and hypervigilance are factors which possibly enable the transformation from episodic into chronic migraine. To elucidate this assumption, attentional parameters were measured by recording contingent negative variation (CNV) and correlated with the individual duration of migraine disease. PATIENTS AND METHODS: A total of 28 patients (episodic migraine with or without aura) were compared with 16 healthy controls. CNV analysis included amplitude and habituation calculation. Data were correlated with the individual duration of the migraine disease. The migraine group was divided into two groups based on a median split (short vs long lasting) which were compared by t-tests. RESULTS: Migraine patients produce higher CNV amplitudes than controls. Moreover, migraineurs showed dishabituation while habituation was seen in controls. There was a Pearson correlation coefficient of r = -0.767 between duration of disease and early component of CNV. Patients with long-lasting disease showed lower dishabituation by a higher intercept than patients with short-lasting disease. CONCLUSION: The correlation between duration of disease and attentional parameters and the changing dishabituation can be interpreted as an enhancement in preactivation level in patients with long lasting migraine. Maybe this change is a prerequisite for transformation into chronic migraine. PMID- 15756582 TI - Spatial variability of leaf wetness duration in different crop canopies. AB - The spatial variability of leaf wetness duration (LWD) was evaluated in four different height-structure crop canopies: apple, coffee, maize, and grape. LWD measurements were made using painted flat plate, printed-circuit wetness sensors deployed in different positions above and inside the crops, with inclination angles ranging from 30 to 45 degrees. For apple trees, the sensors were installed in 12 east-west positions: 4 at each of the top (3.3 m), middle (2.1 m), and bottom (1.1 m) levels. For young coffee plants (80 cm tall), four sensors were installed close to the leaves at heights of 20, 40, 60, and 80 cm. For the maize and grape crops, LWD sensors were installed in two positions, one just below the canopy top and another inside the canopy. Adjacent to each experiment, LWD was measured above nearby mowed turfgrass with the same kind of flat plate sensor, deployed at 30 cm and between 30 and 45 degrees. We found average LWD varied by canopy position for apple and maize (P<0.05). In these cases, LWD was longer at the top, particularly when dew was the source of wetness. For grapes, cultivated in a hedgerow system and for young coffee plants, average LWD did not differ between the top and inside the canopy. The comparison by geometric mean regression analysis between crop and turfgrass LWD measurements showed that sensors at 30 cm over turfgrass provided quite accurate estimates of LWD at the top of the crops, despite large differences in crop height and structure, but poorer estimates for wetness within leaf canopies. PMID- 15756583 TI - Relative importance of fertiliser addition to plants and exclusion of predators for aphid growth in the field. AB - Herbivore dynamics and community structure are influenced both by plant quality and the actions of natural enemies. A factorial experiment manipulating both higher and lower trophic levels was designed to explore the determinants of colony growth of the aphid Aphis jacobaeae, a specialist herbivore on ragwort Senecio jacobaea. Potential plant quality was manipulated by regular addition of NPK-fertiliser and predator pressure was reduced by interception traps; the experiment was carried out at two sites. The size and persistence of aphid colonies were measured. Fertiliser addition affected plant growth in only one site, but never had a measurable effect on aphid colony growth. In both habitats the action of insect predators dominated, imposing strong and negative effects on aphid colony performance. Ants were left unmanipulated in both sites and their performance on the aphid colonies did not significantly differ between sites or between treatments. Our results suggest that, at least for aphid herbivores on S. jacobaea, the action of generalist insect predators appears to be the dominant factor affecting colony performance and can under certain conditions even improve plant productivity. PMID- 15756584 TI - Effects of hydration with salt repletion on renal toxicity of conventional amphotericin B empirical therapy: a prospective study in patients with hematological malignancies. AB - OBJECTIVE: Several studies have suggested that hydration and sodium load might reduce nephrotoxicity related to amphotericin B-deoxycholate (AmB-d). However, a schedule of these nephroprotective measures has not been standardized until now. A protocol of hydration and electrolyte supplementation was used prospectively in patients with hematological malignancies receiving empirical AmB-d treatment to evaluate its effect on AmB-d-related renal toxicity. PATIENTS AND METHODS: A total of 77 consecutive patients received AmB-d (1 mg/kg per day) in association with an initial intravenous hydration of at least 1 l/m2 body surface, containing at least 1 l of 0.9% saline daily. Hydration was increased when serum creatinine levels showed a 20% increase from baseline. Serum electrolytes were replaced when indicated. RESULTS: The median duration of AmB-d therapy was 14 days. The mean intravenous hydration and the mean diuresis were 1530 and 1970 ml/m2 of body surface per day, respectively. Overall, 55 patients (71.4%) received a mean of 18.5 days of therapy without dose-limiting adverse events. Despite significant increases in mean creatinine serum levels and decreases in mean creatinine clearance observed early in the whole population, in only six patients (7.8%) was therapy discontinued due to renal failure, which always recovered after treatment discontinuation. In eight patients (10.4%) therapy was stopped due to infusion related side effects. Seven patients died while under antifungal therapy without relevant signs of AmB-d-associated toxicity. CONCLUSIONS: Our prospective experience confirms that adequate hydration (about 1500 ml/m2 of body surface) and careful electrolyte supplementation are simple measures able to contain nephrotoxicity and to permit adequate antifungal therapy at least in the empirical setting. PMID- 15756585 TI - The association between self-reported physical activity and prevalence of depression and anxiety disorder in long-term survivors of testicular cancer and men in a general population sample. AB - GOALS OF WORK: Physical activity is claimed to have a beneficial influence on a person's mental health, though its impact may be different in cancer patients and individuals in the general population. The objective of this cross-sectional study was (1) to estimate the associations between the level of self-reported physical activity as a lifestyle factor (LPA) and the prevalence of depression and anxiety disorder in testicular cancer survivors (TCSs) and in men of similar age from the general population (GenPop), and (2) to examine whether these associations differ in the two groups. PATIENTS AND METHOD: A total of 1260 TCSs and 20,207 men from the GenPop completed a questionnaire that assessed LPA, and depression and anxiety disorder on the Hospital Anxiety and Depression Scale (HADS). MAIN RESULTS: The prevalence of HADS-defined depression was lower among those who were physically active than in those who were physically inactive (TCSs 9% vs 17%, P<0.001; GenPop 8% vs 15%, P<0.001). Among physically inactive TCSs there was a trend towards higher a prevalence of HADS-defined anxiety disorder compared to physically active TCSs (P=0.07). In the GenPop this difference was statistically significant (P<0.001). Multivariate analysis confirmed the association between LPA and HADS-defined depression in both the TCSs [adjusted odds ratio (aOR)=0.56; 95% confidence interval (CI) (0.31, 1.02)] and the GenPop [aOR=0.58; 95% CI (0.51, 0.65)], but not the association between LPA and HADS defined anxiety disorder. The associations between LPA and HADS-defined depression and HADS-defined anxiety disorder did not differ between the TCSs and the GenPop. CONCLUSION: In both the TCSs and the GenPop, the prevalence of HADS defined depression was higher among those who were physically inactive than among those who were physically active, with no intergroup difference. There was no association between LPA and prevalence of HADS-defined anxiety disorder in either of the groups after adjusting for background variables. Prospective studies are needed to allow causal inferences to be drawn. PMID- 15756586 TI - Formation of an alphaCP1-KH3 complex with UC-rich RNA. AB - The alphaCP family of proteins [also known as poly(C)-binding or heterogeneous nuclear ribonucleoprotein E proteins] are involved in the regulation of messenger RNA (mRNA) stability and translational efficiency. They bind via their triple heterologous nuclear ribonucleoprotein K homology (KH) domain structures to C rich mRNA, and are thought to interact with other mRNA-binding proteins as well as provide direct nuclease protection. In particular, alphaCP1 and alphaCP2 have been shown to bind to a specific region of androgen receptor (AR) mRNA, resulting in its increased stability. The roles of each of the KH motifs in the binding affinity and the specificity is not yet understood. We report the beginning of a systematic study of each of the alphaCP KH domains, with the cloning and expression of alphaCP1-KH2 and alphaCP1-KH3. We report the ability of alphaCP1 KH3, but not alphaCP1-KH2, to bind the target AR mRNA sequence using an RNA electrophoretic mobility gel shift assay. We also report the preparation of an alphaCP1-KH3/AR mRNA complex for structural studies. (1)H-(15)N heteronuclear single quantum correlation NMR spectra of (15)N-labelled alphaCP1-KH3 verified the integrity and good solution behaviour of the purified domain. The titration of the 11-nucleotide RNA target sequence from AR mRNA resulted in a rearrangement of the (1)H-(15)N correlations, demonstrating the complete binding of the protein to form a homogeneous protein/RNA complex suitable for future structural studies. PMID- 15756587 TI - Apparent charge of binding site in ion-translocating enzymes: kinetic impact. AB - Recently, we presented a general scope for the nonlinear electrical properties of enzymes E which catalyze translocation of a substrate S with charge number z(S) through lipid membranes (Boyd et al. J. Membr. Biol. 195:1-12, 2003). In this study, the voltage sensitivity of the enzymatic reaction cycle has been assigned to one predominant reversible reaction step, i.e. the reorientation of either E or ES in the electric field, leaving the reorientation of the alternate state (ES or E) electroneutral, respectively. With this simplification, the steady-state current-voltage relationships (IV) assumed saturation kinetics like in Michaelis Menten systems. Here, we introduce an apparent charge number z(E) of the unoccupied binding site of the enzyme, which accounts for the impact of all charged residues in the vicinity of the physical binding site. With this more realistic concept, the occupied binding site assumes an apparent charge of z(ES) = z(E) + z(S), and IV does not saturate any more in general, but exponentially approaches infinite or zero current for large voltage displacements from equilibrium. These nonlinear characteristics are presented here explicitly. They are qualitatively explained in a mechanistic way, and are illustrated by simple examples. We also demonstrate that the correct determination of the model parameters from experimental data is still possible after incorporating z(E) and its corollaries into the previous model of enzyme-mediated ion translocation. PMID- 15756588 TI - Computing numerically the access resistance of a pore. AB - The access resistance (AR) of a channel is an important component of the conductance of ion channels, particularly in wide and short channels, where it accounts for a substantial fraction of the total resistance to the movement of ions. The AR is usually calculated by using a classical and simple expression derived by Hall from electrostatics (J.E. Hall 1975 J. Gen. Phys. 66:531-532), though other expressions, both analytical and numerical, have been proposed. Here we report some numerical results for the AR of a channel obtained by solving the Poisson-Nernst-Planck equations at the entrance of a circular pore. Agreement is found between numerical calculations and analytical results from Hall's equation for uncharged pores in neutral membranes. However, for channels embedded in charged membranes, Hall's expression overestimates the AR, which is much lower and can even be neglected in some cases. The weak dependence of AR on the pore radius for charged membranes at low salt concentration can be exploited to separate the channel and the access contributions to the measured conductance. PMID- 15756589 TI - Amplification of NOTCH1 and ABL1 gene loci is a frequent aberration in enteropathy-type T-cell lymphoma. AB - We have shown previously that amplification of chromosomal region 9q34 is the most frequent aberration in enteropathy-type T-cell lymphoma (ETL). To determine the minimum amplified 9q34 region and identify possible candidate gene(s), we performed a detailed microsatellite screening and quantitative real-time PCR (QPCR) on 26 ETL cases. Microsatellite analysis revealed allelic imbalance in both ABL1 and NOTCH1 gene loci (microsatellites D9S290-D9S1847 and D9S158 flanking the former and latter genes, respectively) localized in the band 9q34. The results were confirmed by TaqMan-based QPCR showing amplification of ABL1 and NOTCH1 exons in 50% and 65% of cases, respectively. Amplifications of the NOTCH1 gene were more frequent than of the ABL1 gene; moreover, the analyzed NOTCH1 exon consistently displayed higher levels of amplification than ABL1 coding sequences. From 9q34 known genes, NOTCH1 could thus be the primary target of genomic DNA amplification in ETL. PMID- 15756590 TI - A case of mucoepidermoid carcinoma with melanin pigmentation manifested in the palate. AB - This paper describes the first documented case of mucoepidermoid carcinoma (MEC) with melanin pigmentation manifested in the palate. Histopathological sections showed a neoplasm composed of epidermoid, mucous-producing and intermediate cells. Numerous large cells contained dark pigmented materials. Fontana Masson staining revealed dendritic melanocytes and melanin granules. HMB-45, Melan A and S-100 protein were all positive for melanocytes. Histopathological examination was not typical for malignant melanoma; the lesion was diagnosed as a low-grade MEC with melanin pigmentation. PMID- 15756591 TI - Expression of glycodelin A in decidual tissue of preeclamptic, HELLP and intrauterine growth-restricted pregnancies. AB - During pregnancy, the placenta produces a variety of proteins that are responsible for the establishment of the foeto-maternal tolerance and circulation. The aim of this study was to investigate the expression of glycodelin A (formerly named PP14) in decidual tissue of placentas with intrauterine growth restriction (IUGR), preeclamptic patients, hemolysis, elevated liver, low-platelet (HELLP) patients and normal decidual tissue. Slides of paraffin-embedded decidual tissue of patients with IUGR, preeclamptic patients, HELLP patients and normal-term placentas were incubated with either polyclonal or monoclonal antibodies against glycodelin A. Staining reaction was performed with the ABC reagent. Intensity of immunohistochemical reaction on the slides was analysed using a semi-quantitative score. In addition, expression of glycodelin mRNA was analysed by in situ hybridisation. Expression of glycodelin A was significantly reduced in decidual cells of placentas with IUGR and HELLP, as investigated with both monoclonal and polyclonal antibodies and in situ hybridisation. However, preeclamptic decidual tissue showed no significantly different expression of intensity of glycodelin mRNA compared with normal placental tissue controls. A reduced expression of glycodelin A by decidual cells seems to be related to IUGR and HELLP. Therefore, glycodelin A might play an important role in the pathogeneses of these diseases. PMID- 15756592 TI - Gastrointestinal stromal tumor of the pancreas: case report with documentation of KIT gene mutation. PMID- 15756593 TI - Immunohistochemical localization of subtilisin/kexin-like proprotein convertases in human atherosclerosis. AB - Integrins are heterodimeric alpha/beta receptors that link the cytoskeleton with the extracellular matrix, thereby regulating several cell functions important in atherosclerosis. In vitro, the subtilisin/kexin-like proprotein convertases (PCs), namely PC5 and furin, have been shown to be responsible for the endoproteolytic activation of the alpha(v) integrin subunit. Based on their cleavage activity, these PCs are potential targets in atherosclerosis. In the present study, we investigated the localization of furin and PC5 in different stages of human atherosclerosis. Immunohistochemical analysis of furin and PC5 revealed their presence in vascular smooth-muscle cells and endothelial cells in atherosclerotic and non-atherosclerotic lesions. However, in the more advanced lesions, furin and PC5 staining was significantly expressed in macrophages/foam cells. In vitro, THP-1 derived macrophages contained furin and PC5, and maturation of monocytes to macrophages was accompanied by enhanced alpha(v)beta3 cell-surface expression. Inhibition of furin/PC5 with the specific pharmacological furin-like PC-inhibitor dec-CMK inhibited alpha(v) endoproteolytic activation but did not abolish alpha(v)beta3 cell-surface expression. This indicates that furin/PC5 is required for alpha(v) endoproteolytic activation but not for alpha(v) routing and sorting to the cell surface. In conclusion, our study demonstrates that furin and PC5 are significantly expressed in mononuclear cells in advanced human atherosclerotic lesions, where they regulate alpha(v) endoproteolytic activation. PMID- 15756594 TI - Expression of ADAM15 in lung carcinomas. AB - ADAM15, a member of the ADAM (a disintegrin and metalloprotease) family, is a membrane protein containing both protease and adhesion domains and may, thus, be involved in tumor invasion and metastasis. The aim of this study was to analyze the expression of ADAM15 and its potential ligand, integrin alpha(v)beta3 (CD51/CD61), in lung carcinoma cell lines and tissues. Most small cell lung carcinomas (SCLCs) and non-SCLC cell lines were ADAM15, alpha(v) and beta3 integrin mRNA positive. Half of the cell lines expressed ADAM15, and three expressed the alpha(v)beta3 heterodimer at the cell surface as shown using flow cytometry. Paraffin sections of pulmonary epithelial tumors, including SCLCs (n=26), squamous cell cancer (SCCs, n=27) and adenocarcinomas (ACs, n=17) were stained with antibodies to the ectosolic and cytosolic domain of ADAM15 and alpha(v)beta3 integrin complex. The results were scored (0-12, according to Remmele's score). Normal epithelial cells of the lung were negative or slightly positive for ADAM15 (score<2). The score was always significantly higher for tumor cells. ACs showed the strongest staining (tumor center; ADAM15ecto; mean+/ SEM; 5.47+/-1.04), whereas SCLCs only showed weak ADAM15 expression (2.67+/-0.42; SCCs: 3.62+/-0.62). Frequently, significantly stronger ADAM15 expression has been shown in tumor cells located at the front of invasion compared with those within solid formations. Overall analysis of all tumor specimens and each tumor type revealed no significant correlation between tumor stage or degree of differentiation and ADAM15 ectosolic or cytosolic domain expression in tumor cells. Both molecules are often co-localized in the same tumor cells in ADAM15- and alpha(v)beta3 integrin-positive carcinomas. In summary, lung carcinoma cell lines and tissues were frequently ADAM15 positive. PMID- 15756595 TI - Chromophobe renal cell carcinoma with microcystic and adenomatous arrangement and pigmentation--a diagnostic pitfall. Morphological, immunohistochemical, ultrastructural and molecular genetic report of 20 cases. AB - We present clinical, morphological, immunohistochemical, ultrastructural and molecular genetic features of 20 cases of a peculiar form of chromophobe renal cell carcinoma (CRCC) with morphology differing from that of conventional CRCC. Microscopically, the typical features of the tumors were microcystic arrangement and formation of adenomatous structures. Microcystic areas were composed of smaller eosinophilic and bigger pale cells having cytological appearance typical of conventional CRCC. Cytological features of the adenomatous structures were mostly different from those of conventional CRCC. They had a typical columnar arrangement with nuclei positioned at the base of the glandular structures and a small amount of a deeply eosinophilic cytoplasm often endowed with brush border facing the lumen of the glands. In addition, all the tumors showed a brown pigmentation. The pigmentation was located mostly extracellularly, where it formed pools of heavy deposits. Microscopic calcifications present in all cases formed psammoma bodies or else the calcifications were more extensive and amorphous in shape. Ultrastructurally, the cells showed features characteristic of CRCC: typical cytoplasmic vesicles were 100-700 nm in size and mitochondria had tubulovesicular, lamellar or circular cristae. Some tumor cells contained dark, variously sized electron-dense pigment granules. Neither melanosomes nor membrane-bound neurosecretory granules were seen. Using fluorescence in-situ hybridization probes for chromosomes 1, 2, 6, 10, 13, 17 and 21, the tumors revealed massive loss of tested chromosomes typical for conventional CRCC. Monosomy of chromosomes 1, 2, 6, 10, 13 and 21 was found in 100, 36, 91, 82, 82, 82 and 64% of cases, respectively. None of the cases showed mutation of exons 9, 11, 13 and 17 of the c-kit gene. The important feature of pigmented microcystic chromophobe renal cell carcinoma is a relatively benign biological behavior and the absence of distant metastases and sarcomatoid transformation. PMID- 15756597 TI - TRP channels as new pharmacological targets. PMID- 15756598 TI - A kinetic study of trace element leachability from abandoned-mine-polluted soil treated with SS-MSW compost and red mud. Comparison with results from sequential extraction. AB - The effect of adding treated red mud, a by-product of alumina production, to soil polluted by an abandoned mine and characterised by high concentrations of heavy metals, relatively low reaction grade, and low organic carbon content, was investigated. Also studied was addition of both red mud and compost (produced from source-separated municipal solid waste)--the synergistic action of red mud and compost could be exploited to achieve both metal trapping and an increase in organic carbon content. Leaching batch tests were performed on four different systems: soil, soil and treated red mud, soil and compost, soil and compost plus treated red mud. Dilute sulfuric acid and EDTA solution (liquid/solid ratio 10:1) were used in the tests--sulfuric acid to "mimic" acid rain and EDTA in accordance with general methods for estimating "plant-available" metals. Sequential extraction was also applied to the same samples. The use of relatively non specific extractant reagents in the leaching tests led to a kinetic approach (already proposed in literature), because measurements of trace elements extracted at equilibrium cannot be related to their speciation. Comparison of information obtainable by the kinetic approach to evaluation of data from leaching tests with results from sequential extraction enabled evaluation whether the "kinetic fractionation method", a relatively rapid and simple procedure, furnishes adequate information about the mobility and bioavailability of trace elements. Especially interesting results were obtained for Mn, Zn, and Ni, present in large amounts in the soil studied--their leachability was significantly reduced by addition of red mud and compost, suggesting interesting perspectives in soil-remediation activity. PMID- 15756596 TI - Impact of interleukin-12, oxidative burst, and iNOS on the survival of murine fecal peritonitis. AB - Abdominal sepsis due to secondary fecal peritonitis following anastomosis insufficiency is a rare but life threatening complication of colorectal surgery. The induction of IFN-gamma by IL-12 is believed to play a key role in sepsis as it promotes antibacterial effector mechanisms such as oxidative burst or nitric oxide induction. The impact of gene deficiency for IL-12 (IL-12p40 KO), oxidative burst (p47(phox) KO), or NO induction (iNOS KO) on the outcome of fecal peritonitis was characterized using the murine Colon Ascendens Stent Peritonitis model (CASP). In the IL-12p40 KO model, 3 and 12 h after surgery, serum cytokine levels of IL-1beta, TNF, IL-18, and IL-10 were analyzed. Expression of IL-1beta, IL-10, IP-10, and MIP-1alpha was measured in lung and liver by RNAse Protection Assay. IL-12p40 and iNOS-deficient mice exhibited a significantly higher susceptibility to CASP as compared to the controls, whereas no significant difference was observed in p47(phox) KO mice. Absence of IL-12 resulted in delayed expression of proinflammatory cytokines and chemokines in both the liver and the lung, and was associated with significant reduction of IL-1beta levels in the serum 12 h after CASP. IL-12 and iNOS possess protective functions in fecal murine peritonitis. Surprisingly, no significant contribution of oxidative burst to the immune response was observed. Overall, these findings suggest that IL-12 deficiency causes a profound delay of the immune response after polymicrobial challenge resulting in significantly increased susceptibility in the CASP model. PMID- 15756600 TI - Quantitative analysis of hydrogen peroxide by 1H NMR spectroscopy. AB - A technique utilizing 1H NMR spectroscopy has been developed to measure the concentration of hydrogen peroxide from 10(-3) to 10 M. Hydrogen peroxide produces a peak at around 10-11 ppm, depending upon the interaction between solvent molecules and hydrogen peroxide molecules. The intensity of this peak can be monitored once every 30 s, enabling the measurement of changes in hydrogen peroxide concentration as a function of time. 1H NMR has several advantages over other techniques: (1) applicability to a broad range of solvents, (2) ability to quantify hydrogen peroxide rapidly, and (3) ability to follow reactions forming and/or consuming hydrogen peroxide as a function of time. As an example, this analytical technique has been used to measure the concentration of hydrogen peroxide as a function of time in a study of hydrogen peroxide decomposition catalyzed by iron(III) tetrakispentafluorophenyl porphyrin. PMID- 15756599 TI - Detection of N-monomethyl-lysine generated by metabolic transmethylation. AB - Administration of radiolabelled deprenyl to rats resulted in the urinary elimination of a (14)C-labelled N(epsilon)-monomethyl-lysine. An increased level of N(epsilon)-monomethyl-lysine was found following an oral dose of another drug, also containing an N-methyl group. The urine sample was treated with 9 fluorenylmethoxycarbonyl chloride and then subjected to high-performance liquid chromatography (HPLC); the radioactive fraction was identified as N(epsilon) monomethyl-lysine by using HPLC-MS in electrospray mode. Identification of N(epsilon)-monomethyl-lysine in the radioactive fraction gives experimental proof of transmethylation from a well-known drug to an endogenous compound. PMID- 15756601 TI - Clean up of phenylurea herbicides in plant sample extracts using molecularly imprinted polymers. AB - Two different molecularly imprinted polymers (MIPs) were prepared by precipitation polymerization using linuron or isoproturon (phenylurea herbicides) as templates and trifluormethacrylic acid as functional monomer. These materials were used as selective sorbents in the development of molecularly imprinted solid phase extraction (MISPE) procedures for the determination of several phenylurea herbicides (fenuron, metoxuron, chlortoluron, isoproturon, metobromuron, and linuron) in plant samples extracts. The MISPE procedures were fully optimized and applied to the clean up of selected phenylurea herbicides in carrot, potato, corn, and pea sample extracts and finally determined by HPLC-UV at 244 nm. Although a high degree of clean up was obtained, a decrease of the MIP recognition capabilities was observed in subsequent runs. Thus, a previous clean up protocol based on the use of a non-imprinted polymer was used to prevent the loss of MIP performance and to ease the removal of interferences. Following this procedure, namely two-step MISPE, matrix compounds were almost completely removed by the non-imprinted polymer retaining the ability of MIPs to selectively rebind target analytes unaltered. The developed MISPE procedures allowed the screening of phenylurea herbicides in plant samples at concentration levels required by established European maximum residue limits. PMID- 15756602 TI - Sandwich immunoassay for determination of vitellogenin in golden grey mullet (Liza aurata) serum as a field exposure biomarker. AB - Vitellogenin (VTG) is a protein produced in the liver of oviparous animals in response to oestrogens. Abnormal production of VTG by males, therefore, is used as a biological indicator of exposure to xeno-oestrogens. In this study, a sandwich-ELISA for measuring VTG in Liza aurata (golden grey mullet) was developed and validated. Plasma VTG was purified from 17beta-oestradiol-injected immature individuals of mullet, by size-exclusion and ion-exchange chromatography. Polyclonal antibodies against VTG were raised in rabbits. A sensitive immunoassay was developed for measurement of vitellogenin in L. aurata serum, reaching a quantification limit of 0.01 microg mL(-1) and a dynamic range from 0.02 to 2 microg mL(-1). The assay is specific, because high levels (>100 microg mL(-1)) of carp (Cyprinus carpio), goldfish (Carassius auratus), tilapia (Oreochromis niloticus), tench (Tinca tinca), rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and frog (Rana perezi) purified VTG, give negligible responses. The assay was used to analyse plasma samples from wild mullet. PMID- 15756603 TI - Using enzymes isolated from diverse sources to determine metal ion cofactors. AB - Oxidoreductases and hydrolases isolated from different sources (horseradish and peanut peroxidases, alcohol dehydrogenases from baker's yeast and horse liver, and alkaline phosphatases from Escherichia coli, chicken and seal intestine) were used to determine their metal ion cofactors: Fe(III), Zn(II) and Mg(II), respectively. Studying the effects of the metal ion cofactors on the catalytic activity of the enzymes of different origin showed that the extent of their inhibition, activation, or reactivation of their apoenzymes depended on the structure and accessibility of the enzyme active site, which varies among the biocatalysts isolated from different sources. The developed procedures are based on the inhibiting (Zn(II)) or activating (Mg(II)) effects of the metal ions on the catalytic activity of the enzymes, or on reactivating effects (Fe(III) and Zn(II)) on the apoenzymes. The procedures are characterized by high sensitivity and selectivity; the detection limits of Fe(III) using horseradish peroxidase, Zn(II) using alcohol dehydrogenase from baker's yeast, alkaline phosphatase from seal intestine and its apoenzyme, and Mg(II) using alkaline phosphatase from chicken intestine equal 10 ng L(-1), 20 ng L(-1), 3 microg L(-1), 8 microg L(-1) and 0.2 microg L(-1), respectively. PMID- 15756604 TI - Heat shock protein 70/MAGE-3 fusion protein vaccine can enhance cellular and humoral immune responses to MAGE-3 in vivo. AB - MAGE-3, a member of melanoma antigen (MAGE) gene family, is recognized as an ideal candidate for tumor vaccine because it is expressed in a significant proportion of tumors of various histological types and can induce antigen specific immune response in vivo. There is now substantial evidence that heat shock proteins HSPs isolated from cancer cells and virus-infected cells can be used as vaccines to produce cancer-specific or virus-specific immunity. In this research, we investigated whether M. tuberculosis HSP70 can be used as vehicle to elicit immune response to its accompanying MAGE-3 protein. A recombinant protein expression vector was constructed that permitted the production of fusion protein linking amino acids 195-314 of MAGE-3 to the C terminus of HSP70. We found that HSP70-MAGE-3 fusion protein can elicit stronger cellular and humoral immune responses against MAGE-3 expressing murine tumor than those elicited by MAGE-3 protein in vivo, which resulted in potent antitumor immunity against MAGE-3 expressing tumors. Covalent linkage of HSP70 to MAGE-3 was necessary to elicit immune response to MAGE-3. These results indicate that linkage of HSP70 to MAGE-3 enhanced immune responses to MAGE-3 in vivo and HSP70 can be exploited to enhance the cellular and humoral immune responses against any attached tumor-specific antigens. PMID- 15756607 TI - Aneurysmal subarachnoid haemorrhage in the first year of life. PMID- 15756605 TI - Complex tibial plateau fractures treated with Ilizarov external fixator with or without minimal internal fixation. AB - We treated 30 tibial plateau fractures (Schatzker Type VI) in 29 patients, with a mean age of 41.4 (20-76) years, with the Ilizarov fixator. In 18 fractures, we combined the treatment with minimal internal fixation. All fractures were the result of high-energy trauma, and 20 patients had associated injuries. Twenty eight fractures were available for follow-up after 27 (16-36) months. Using The Knee Society clinical rating system, 18 knees were rated as excellent, seven as good, one as fair, and two as poor. There was a direct correlation between the presence of associated injuries and the final outcome. The most significant concomitant injuries were distal femoral fractures and extensive soft-tissue injury. This study emphasizes the clinical success and low morbidity associated with the use of external fixation and minimal internal fixation. PMID- 15756608 TI - Prevalence and clinical significance of superficial abdominal reflex abnormalities in idiopathic scoliosis. AB - To determine prevalence and significance of abnormal superficial abdominal reflexes (SARs) in idiopathic scoliosis. Study of 73 patients with presumed idiopathic scoliosis referred for magnetic resonance imaging (MRI), either as a routine pre-operative assessment (n=42) or because of abnormal symptoms or neurological signs (n=31). All patients were examined prior to magnetic resonance imaging (MRI), and the presence of abnormal SARs was noted. All patients then underwent MRI of the whole spine from the foramen magnum to the sacrum. The presence of Chiari 1 malformation and syrinx was recorded. The study group consisted of 11 males and 62 females with a mean age at time of MRI of 18 years (range 5-51 years) and a mean Cobb angle of 48 degrees (range 10-104 degrees). Abnormality of the SARs was recorded in eight cases (prevalence 11%). An abnormal MRI study was recorded in nine cases (12.3%), all patients having a syrinx and four having in addition, a Chiari 1 malformation. Of the patients with abnormal SARs, only 2 (25%) had an abnormal MRI study; 1 had unilateral absence of the reflexes whereas the other had complete absence of SARs. Of patients referred for MRI as a routine pre-operative assessment, 5 (11.6%) had an abnormal MRI study. In patients with idiopathic scoliosis, abnormality of the SARs was recorded in 11% of cases. Unilateral absence was present in one case only and was associated with the presence of syrinx. Other patterns of abnormality were not a useful indicator of underlying cord abnormality. PMID- 15756610 TI - New ideas about chronic tendinosis and the use of strain-gauges inside the shoulder. PMID- 15756609 TI - Internal derangements of the knee associated with patellofemoral joint degeneration. AB - The objective of the present study was to evaluate whether internal derangements of the knee joint are associated with patellofemoral joint (PFJ) degeneration. Data were collected prospectively from 1,000 consecutive knee arthroscopies. Chondral lesions were observed in 854 patients, and these patients were included in this study. Patients' details (age, sex, duration of symptoms, injuries, and possible mechanism of injury), operative details (types and number of portals, equipment used), intra-articular findings (articular, meniscal and synovial lesions, and stability characteristics) and procedures performed were recorded. Articular lesions were noted on anatomic articular maps of the different functional zones, using a system which presaged the current ICRS system. Using this zoning system, it was easy for the authors to separate the patients with isolated patellofemoral degeneration and arthritis and to do the statistical analysis. Increased incidence of isolated patellofemoral degeneration was noted in patients with synovial shelves, in comparison with patients without shelves (24.7% vs. 15.5%, respectively; P=0.001). Patients with severe patellofemoral maltracking were found to have increased incidence of isolated patellofemoral degeneration in comparison with patients without patella maltracking (64.7% vs. 18% respectively; P<0.001). In conclusion, patellofemoral maltracking and synovial shelves of the knee are highly associated with PFJ degeneration. On the contrary, meniscal tears and ligamentous injuries are associated mainly with degeneration of other compartments of the knee joint and not the PFJ. PMID- 15756611 TI - Neovascularisation in chronic painful patellar tendinosis--promising results after sclerosing neovessels outside the tendon challenge the need for surgery. AB - Sclerosing injections targeting neovascularisation have been demonstrated to give promising clinical results in patients with chronic painful Achilles tendinosis. In this study, fifteen elite or recreational athletes (12 men and three women) with the diagnosis patellar tendinosis/Jumper's knee in 15 patellar tendons were included. All the patients had a long duration of pain symptoms (mean = 23 months) from the patellar tendon, and ultra-sonography + colour doppler examination showed structural tendon changes with hypo-echoic areas and a neovascularisation, corresponding to the painful area. The patients were treated with ultrasound and colour doppler-guided injections of the sclerosing substance Polidocanol, targeting the area with neovascularisation. At follow-up (mean = 6 months) after a mean amount of three treatments, there was a good clinical result in 12/15 tendons. The patients were back to their previous (before injury) sport activity level, and the amount of pain recorded on a VAS-scale had decreased significantly (VAS from 81 to 10). Our findings indicate that treatment with sclerosing injections, targeting the area with neovessels in patellar tendinosis, has the potential to cure the pain in the tendons and also allow the patients to go back to full patellar-tendon loading activity. PMID- 15756613 TI - The effect of femoral attachment location on anterior cruciate ligament reconstruction: graft tension patterns and restoration of normal anterior posterior laxity patterns. AB - The issue of the best place to attach an anterior cruciate ligament graft to the femur is controversial, and different anatomic or isometric points have been recommended. It was hypothesised that one attachment site could be identified that would be best for restoring normal anterior-posterior laxity throughout the range of knee flexion. It was also hypothesised that these different attachment sites would cause different graft tension patterns during knee flexion. Using six cadaver knees, an isometric point was found 3 mm distal to the posterior edge of Blumensaat's line, at the 10:30-11:00 o'clock position in right knees, at the antero-proximal edge of the anatomic ACL attachment. Anterior-posterior laxity was measured at +/-150 N draw force at 20-120 degrees flexion with the knee intact and after anterior cruciate ligament transection. The graft was placed at the isometric point, and AP laxity was restored to normal at 20 degrees flexion, then measured at other angles. Graft tension was measured throughout, and also during passive flexion-extension. This was repeated for four other graft positions around the isometric point in every knee. Laxity was restored best by grafts tensioned to a mean of 9 +/- 14 N, positioned isometrically and 3 mm posterior to the isometric point. Their tension remained low until terminal extension. Grafts 3 mm anterior to the isometric point caused significant overconstraint, and had higher tension beyond 80 degrees knee flexion. Small changes in attachment site had large effects on laxity and tension patterns. These results support an isometric/posterior anatomic femoral graft attachment, which restored knee laxity to normal from 20 to 120 degrees flexion and did not induce high graft tension as the knee flexed. Grafts attached to the roof of the intercondylar notch caused overconstraint and higher tension in the flexed knee. PMID- 15756612 TI - Anterior cruciate ligament reconstruction with double-looped semitendinosus and gracilis tendon graft directly fixed to cortical bone: 5-year results. AB - Forty-three patients who had undergone an anterior cruciate ligament (ACL) reconstruction using a doubled semitendinosus and gracilis graft were prospectively reviewed at 5-year follow-up. All had suffered subacute or chronic tears of the ligament. At surgery, the femoral tunnel was drilled first through the antero-medial portal. The correct position of the femoral and tibial guide wire was checked fluoroscopically. A cortical fixation to the bone was achieved in the femur with a Mitek anchor, directly passing the two tendons in the slot of the anchor, and in the tibia with an RCI screw, supplemented with a spiked washer and bicortical screw. Rehabilitation was aggressive, controlled and without braces. The International Knee Documentation Committee (IKDC) form, KT-1000 arthrometer, and Cybex dynamometer were employed for clinical evaluation. A radiographic study was also performed. At the 5-year follow-up all the patients had recovered full range of motion and 2% of them complained of pain during light sports activities. Four patients (9.5%) reported giving-way symptoms. The KT-1000 side-to-side difference was on average 2.1 mm at 30 lb, and 68% of the knees were within 2 mm. The final IKDC score showed 90% satisfactory results. There was no difference between the 2-year and 5-year evaluations in terms of stability. Extensor and flexor muscle strength recovery was almost complete (maximum deficit 5%). Radiographic study showed a tunnel widening in 32% of the femurs and 40% of the tibias. A correlation was found between the incidence of tibial tunnel widening and the distance of the RCI screw from the joint (the closer the screw to the joint, the lower the incidence of widening). In conclusion, we can state that, using a four-strand hamstring graft and a cortical fixation at both ends, we were able to achieve satisfactory 5-year results in 90% of the patients. PMID- 15756615 TI - The effect of anterior cruciate ligament reconstruction on lower extremity relative phase dynamics during walking and running. AB - The purpose of this investigation was to use relative phase dynamics to evaluate gait in individuals with a reconstructed anterior cruciate ligament (ACL) during walking and running. Relative phase dynamics can describe the coordination strategies between the interacting segments at the lower extremity. Ten subjects who had undergone ACL reconstruction using the central third of their patellar tendon and ten healthy controls walked and ran on a treadmill at a self-selected pace. Relative phase dynamics were calculated for the foot-shank and shank-thigh coordinative relationships. Statistical differences between the groups were noted for the foot-shank relationship (p < 0.05) during both walking and running and for the shank-thigh relationship (p < 0.05) during walking. Our results indicate that current ACL reconstructive techniques may result in altered relative phase dynamics. These changes in relative phase dynamics could be related to a loss of sensory information about joint position and velocity that is typically provided by the intact ACL. Additionally, relative phase adaptations could be a learned response from the early stages of postsurgical rehabilitation. Relative phase dynamics provide quantitative information about the dynamic status of the ACL reconstructed knee that cannot be gained from the conventional time-series evaluation of gait analysis data. Relative phase dynamics measures should supplement the conventional gait analysis measures that are used today for the clinical evaluation of the functional dynamic stability of the reconstructed knee. The examination of relative phase dynamics could be clinically important for the quantification of new ACL surgical interventions and of patient performance at various stages of rehabilitation. Further research should incorporate relative phase dynamics to understand the influence of ACL reconstruction on coordination and functional patient outcomes. PMID- 15756614 TI - The load of an implanted graft during and after fixation in anterior cruciate ligament reconstruction. AB - Although the importance of initial graft load has been discussed in the literature, it has not been confirmed whether a surgeon can provide the desired load to an implanted graft in anterior cruciate ligament reconstruction. The purpose of this study was to compare the set force (initial load given to the graft before fixation) and residual load in the implanted graft using three different fixation techniques. A total of ninety porcine knees were tested using bone-patellar tendon-bone autograft. Each bone-tendon-bone autograft was fixed to the tibia with either the interference-fit screw, fixation post, or button technique. Graft load was monitored during fixation procedures, and for 10 min after fixation. Residual graft load with each fixation technique exhibited unique features. Highest graft load was obtained by the interference-screw fixation technique; however, the graft was usually over-loaded beyond the intended set force with this technique. In the fixation-post technique, the load of the graft increased while the screw was retightened. The graft load in the button-fixation technique was low, probably due to slight slippage of the button. When the graft was fixed under maximum manual tension, the graft loads at completion of fixation for the interference fit screw, fixation post, and button techniques were 116.3, 54.2, and 25.9 N respectively. PMID- 15756616 TI - Clinical outcome of arthroscopic reduction and suture for displaced acute and chronic tibial spine fractures. AB - This paper reports the clinical outcome of the arthroscopic reduction and pull out suture technique in acute and chronic displaced tibial spine anterior cruciate ligament (ACL) avulsion fractures. Between April 1997 and December 2000, 14 patients received an arthroscopic reduction and pull-out suturing of displaced tibial spine fractures (ACL avulsion fractures of tibia). Of 14 cases, ten were acute fractures and four were chronic nonunion fractures, in which all patients showed extension limitation. The mean follow-up period was 51 months (ranging from 30 to 80 months). At final follow-up, review of range of motion, Lachman test, anterior drawer test, KT-2000 arthrometer, Lysholm knee score, and Hospital for Special Surgery (HSS) score were evaluated. Compared to conventional pull-out suturing, several key modifications to surgical techniques were used. In all 14 patients, radiological bony union was detected at mean 12.3 weeks (range, 8-16 weeks) after surgery. All patients were able to return to their preinjury activity and sports level. At final follow-up, full range of motion was achieved in all patients. Anterior draw test, Lachman test, and KT-2000 (less than 3 mm side-to-side) were all negative in 13 patients. One female patient, who was 6 years old at the time of surgery, complained of no subjective instability, but showed Lachman grade I, and 5 mm side-to-side difference in KT-2000. She also revealed 10 degrees difference of genu recurvatum deformity. Two children (including the previously-mentioned 6-year-old female patient) showed leg-length discrepancy of 1 cm-the affected legs being longer-at final follow-up. The mean Lysholm knee scores were 95.6 (range, 92-100) and HSS knee scores were 96.4 (range, 91-100). Arthroscopic reduction with modified pull-out suturing technique in displaced tibial spine ACL avulsion fractures showed excellent union rate for both acute and chronic cases, without instability or extension limitations at minimum two-year follow-up. PMID- 15756617 TI - Primary cartilage lesions and outcome among subjects with patellofemoral pain syndrome. AB - We investigated the association between cartilage lesion and future symptoms in two groups of subjects with patellofemoral pain syndrome (PFPS). Group A consisted of 31 men (mean age 20.9 years, range 19-32) and group B of 28 consecutive patients (12 men and 16 women; mean age 27.8 years, range 15-50) operated on for PFPS. The mean follow-up time was 3.4 (range 1-7) years in group A and 4.9 (range 1-6) years in group B. At follow-up the subjects in group A with severe cartilage lesion (n = 10) reported more knee symptoms as measured by their Kujala score (mean 75.1, SE 3.6 vs. 88.5, SE 2.4) than those with minor cartilage lesion (n = 21) (age and follow-up time-adjusted P < 0.01). In group B the subjects with severe cartilage lesion tended to report more knee symptoms according to their Kujala score (mean 71.0, SE 7.0 vs. 86.1, SE 5.9; age, sex and follow-up time-adjusted P = 0.15) and VAS score (mean 51.8, SE 11.8 vs. 12.7, SE 10.4; adjusted P = 0.04) than the subjects without cartilage lesion. Our study shows that among the patients with PFPS, the subjects with severe cartilage lesion of the patella or femoral trochlea reported more subjective symptoms and functional limitations at follow-up than those without or with small cartilage lesion of the patella or femoral trochlea. PMID- 15756618 TI - Infrapatellar fat pad pressure and volume changes of the anterior compartment during knee motion: possible clinical consequences to the anterior knee pain syndrome. AB - This biomechanical study was performed to measure tissue pressure in the infrapatellar fat pad and the volume changes of the anterior knee compartment during knee flexion-extension motion. Knee motion from 120 degrees of flexion to full extension was simulated on ten fresh frozen human knee specimens (six from males, four from females, average age 44 years) using a hydraulic kinematic simulator (30, 40, and 50 Nm extension moment). Infrapatellar tissue pressure was measured using a closed cell sensor. Infrapatellar volume change in the anterior knee compartment was evaluated subsequent to removal of the fat pad using a water filled bladder. We found a significant increase of the infrapatellar tissue pressure during knee flexion, at flexion angles of <20 degrees and >100 degrees . The average tissue pressure ranged from 343 (+/-223) mbar at 0 degrees to 60 (+/ 64) mbar at 60 degrees of flexion. The smallest volume in the anterior knee compartment was measured at full extension and 120 degrees of flexion, whereas the maximum volume was observed at 50 degrees of flexion. In conclusion, the data suggest a biomechanical function of the infrapatellar fat pad at flexion angles of <20 degrees and >100 degrees , which suggests a role of the infrapatellar fat pad in stabilizing the patella in the extremes of knee motion. PMID- 15756619 TI - A novel technique to measure active tendon forces: application to the subscapularis tendon. AB - This paper describes the design and use of a novel system for quantification of active tendon forces. An arthroscopically implantable force probe (AIFP) was inserted arthroscopically into the subscapularis tendon of the shoulder in vivo. The output response of this device was calibrated in situ with known forces applied to the tendon using an arthroscopic technique. On recovery of motor control following interscalene regional anaesthesia, the force within the tendon during active muscle contraction was measured. The AIFP was then retrieved through an arthroscopic portal. The technique showed that the subscapularis may produce forces of 250 N during maximum internal rotation of the shoulder. PMID- 15756620 TI - Surgical management of chronic, unreduced posterior dislocation of the shoulder. AB - Four patients with chronic posterior dislocation of the shoulder underwent surgical treatment after an average delay of 10 months from injury. They were examined clinically and radiographically at an average follow-up of 32 months. Treatment consisted of subscapularis tendon transfer (McLaughlin procedure) into the humeral defect in one case, transfer of the lesser tuberosity (McLaughlin modified procedure) in two cases, and in the fourth case-a patient with a 19 month missed dislocation and 50% humeral head lesion-a transposition of the coracoid process and conjoined tendon was performed. X-rays and CT scan excluded avascular necrosis or severe post-traumatic arthrosis. All patients achieved complete pain relief without limitation of daily or work activities. A slight limitation of anterior elevation and external rotation was present in all patients. Our results confirm that McLaughlin's original or modified procedure is suggested in cases of chronic, unreduced posterior dislocation of the shoulder (type I according to Randelli). The transposition of the coracoid process is a valid alternative to prosthesis and to autologous or homologous bone graft implants. PMID- 15756627 TI - [Pneumologie 2005 -- future by transition]. PMID- 15756621 TI - Salt-dependent expression of ammonium assimilation genes in the halotolerant yeast, Debaryomyces hansenii. AB - Debaryomyces hansenii is adapted to grow in saline environments, accumulating high intracellular Na(+) concentrations. Determination of the DhGDH1-encoded NADP glutamate dehydrogenase enzymatic activity showed that it increased in a saline environment. Thus, it was proposed that, in order to overcome Na(+) inhibition of enzyme activity, this organism possessed salt-dependent mechanisms which resulted in increased activity of enzymes pertaining to the central metabolic pathways. However, the nature of the mechanisms involved in augmented enzyme activity were not analyzed. To address this matter, we studied the expression of DhGDH1 and DhGLN1 encoding glutamine synthetase, which constitute the central metabolic circuit involved in ammonium assimilation. It was found that: (1) expression of DhGDH1 is increased when D. hansenii is grown in the presence of high NaCl concentrations, while that of DhGLN1 is reduced, (2) DhGDH1 expression in Saccharomyces cerevisiae takes place in a GLN3- and HAP2,3-dependent manner and (3) salt-dependent DhGDH1 and DhGLN1 expression involves mechanisms which are limited to D. hansenii and are not present in S. cerevisiae. Thus, salt-dependent regulation of the genes involved in central metabolic pathways could form part of a strategy leading to the ability to grow under hypersaline conditions. PMID- 15756628 TI - [Lung cancer screening revisited -- are we merely combing a giraffe?]. PMID- 15756630 TI - [Aspiration of dental implant -- reasons, management and prevention]. AB - Aspiration of foreign bodies is extremely rare in adults, but it may occur during the course of dental treatment and become a serious problem for the patient. A case of a 77-year old man with chronic cough after dental treatment and unrecognised aspiration of an implant is presented. The implant remained unidentified for three years as an endobronchial foreign body of the lower lobe bronchus. A bronchoscopic removal failed and surgery was indicated for lobectomy of the right lower lobe because of chronic pneumonia. A review of prevention, signs and symptoms, management and documentation of the complications is presented. PMID- 15756629 TI - [Asthma control with the salmeterol-fluticasone-combination disc compared to standard treatment]. AB - BACKGROUND: The present study aimed to investigate whether a fixed combination of salmeterol and fluticasone (SFC) from a single inhaler provides sufficient asthma control comparable to that achieved with standard treatment (inhaled steroid in a dose of 1,000 mcg BDP- (beclomethasone dipropionate) equivalent plus a LABA and/or theophylline and/or montelukast). PATIENTS AND METHODS: In a prospective, randomised study patients with moderate or severe asthma were either switched to a twice daily inhalation of 50 mcg salmeterol plus 500 mcg fluticasone from the Viani forte 50/500 mcg Diskus (n = 142 patients), or they were maintained on standard treatment (n = 89 patients). If adequate asthma control was achieved after 8 weeks, the dose of the inhaled steroid was reduced by 50 % during weeks 9 to 16. RESULTS: After 8 weeks, 81 % of the patients who had been switched to SFC and 80 % of patients on standard treatment achieved sufficient asthma control. After reducing the ICS dose by 50 %, asthma control remained appropriate in 90 % of SFC-patients, but only in 75 % of patients receiving standard treatment (p = 0.031). In addition, asthma symptoms and use of rescue medication were significantly more stable in SFC patients (p < 0.05). CONCLUSIONS: With the salmeterol fluticasone combination product, patients with moderate asthma can achieve a control of their asthma, which is as good as that after standard treatment. In most SFC patients the fluticasone dosage can be reduced by 50 % without losing asthma control. PMID- 15756631 TI - [Lung cancer screening -- state of the art]. AB - Besides smoking as the main risk factor for lung cancer several occupational risk factors like exposure to asbestos, polycyclic aromatic hydrocarbons, uranium and dust containing nickel or silica have to be considered. Due to lack of effort in smoking prevention and cessation, lung cancer screening is an important issue. A number of pitfalls has to be considered when evaluating the efficacy of screening procedures. In this paper, we summarize the results of the major studies including chest X-ray, sputum cytology and low dose computed tomography. Randomized controlled studies involving low dose CT in about 100 000 subjects are on the way. Around the year 2010 we will be able to define whether or not lung cancer screening including new techniques and standardized algorithms yields a decrease in mortality. If diagnostic algorithms are used which have been applied in published feasibility studies, the mean percentage of invasive diagnostic measures revealing benign lesions is about 34 % and thus below those obtained in, e. g., breast cancer screening trials. PMID- 15756632 TI - [A connection between allergic rhinitis and allergic asthma? The "one-airway-one disease"-hypothesis]. AB - As outlined in part 1, numerous data from immunologic, genetic and epidemiologic studies point to a systemic link between allergic asthma and rhinitis which can be seen as manifestations of a common atopic syndrome. In part 2 clinical manifestations, diagnostics and most importantly therapeutic options effecting on both nasal and bronchial symptoms will be discussed. Allergen avoidance is the first step in therapeutic management of allergic diseases. Specific immunotherapy (SIT), leukotriene modifying compounds (in Germany exclusively Montelukast), and corticosteroids inhibit inflammation in the epithelium of the upper and the lower airways. Although SIT has a widely accepted indication in the treatment of allergic rhinitis, it is just provisionally recommended for the treatment of asthmatic patients. Most recently Montelukast, a potent leukotriene receptor inhibitor, has been approved for the treatment of asthma as well as for allergic rhinitis. Local administration of corticosteroids requires that they be given both nasally and bronchially. Just on the i. v. or oral route corticosteroids may inhibit the allergic inflammation in both compartments. Newly developed IgE inhibitor Omalizumab, which has no approval in Germany yet, has been reported to have similar effects. Thus, various therapeutic options are available to treat asthma and rhinitis at the same time. Furthermore, multilateral clinical efficacy of antiinflammatory drugs support the "One-Airway-One-Disease" hypothesis. PMID- 15756633 TI - [Integrated medical care]. PMID- 15756634 TI - [New developments in the health care system and their impact on pulmonology]. PMID- 15756635 TI - [Athanasius Kirchers' plague treatise of 1658 and his views of the pulmonary plague]. PMID- 15756636 TI - [Prof. Dr. med. Heinrich Matthys on the occasion of his 70th birthday]. PMID- 15756637 TI - Mutations in ABCA12 underlie the severe congenital skin disease harlequin ichthyosis. AB - Harlequin ichthyosis (HI) is the most severe and frequently lethal form of recessive congenital ichthyosis. Although defects in lipid transport, protein phosphatase activity, and differentiation have been described, the genetic basis underlying the clinical and cellular phenotypes of HI has yet to be determined. By use of single-nucleotide-polymorphism chip technology and homozygosity mapping, a common region of homozygosity was observed in five patients with HI in the chromosomal region 2q35. Sequencing of the ABCA12 gene, which maps within the minimal region defined by homozygosity mapping, revealed disease-associated mutations, including large intragenic deletions and frameshift deletions in 11 of the 12 screened individuals with HI. Since HI epidermis displays abnormal lamellar granule formation, ABCA12 may play a critical role in the formation of lamellar granules and the discharge of lipids into the intercellular spaces, which would explain the epidermal barrier defect seen in this disorder. This finding paves the way for early prenatal diagnosis. In addition, functional studies of ABCA12 will lead to a better understanding of epidermal differentiation and barrier formation. PMID- 15756638 TI - Exon array CGH: detection of copy-number changes at the resolution of individual exons in the human genome. AB - The development of high-throughput screening methods such as array-based comparative genome hybridization (array CGH) allows screening of the human genome for copy-number changes. Current array CGH strategies have limits of resolution that make detection of small (less than a few tens of kilobases) gains or losses of genomic DNA difficult to identify. We report here a significant improvement in the resolution of array CGH, with the development of an array platform that utilizes single-stranded DNA array elements to accurately measure copy-number changes of individual exons in the human genome. Using this technology, we screened 31 patient samples across an array containing a total of 162 exons for five disease genes and detected copy-number changes, ranging from whole-gene deletions and duplications to single-exon deletions and duplications, in 100% of the cases. Our data demonstrate that it is possible to screen the human genome for copy-number changes with array CGH at a resolution that is 2 orders of magnitude higher than that previously reported. PMID- 15756639 TI - Impaired Th1 responses in mice deficient in Epstein-Barr virus-induced gene 3 and challenged with physiological doses of Leishmania major. AB - Protection against Leishmania major is dependent on IL-12 release from L. major infected dendritic cells (DC) that induce IFN-gamma-producing Th1/Tc1 cells. IL 27, a novel member of the IL-12 family, is a heterodimer composed of p28 and IL 12p40-related Epstein-Barr virus-induced gene 3 (EBI3), and was shown to be produced by DC. In this study, we utilized EBI3-deficient mice to investigate the role of IL-27 in leishmaniasis using physiological low-dose infections that mimic natural transmissions. Lesions in EBI3(-/-) mice were significantly larger between weeks 3 and 10 post infection, reaching up to approximately threefold increased lesion volumes compared to wild types. In parallel, dermal lesions of EBI3(-/-) mice contained greater parasite numbers, reaching a peak load that was 2-log higher than in C57BL/6 mice. However, lesions in EBI3(-/-) and wild-type mice resolved after 12 weeks. At early time points, the antigen-specific cytokine response in EBI3(-/-) lymph nodes showed increased levels of IL-4, IL-10 and IL 13 and decreased IFN-gamma production. IL-27 production was restricted to the DC population, since the majority of EBI3 expression in lymph nodes of infected mice was found in CD11c(+) cells. In conclusion, our data show that DC-derived IL-27 is critical for the timely initiation of efficient anti-parasite Th1 immunity early in infections. PMID- 15756640 TI - Investigation of potential RNA bulge stabilizing elements. AB - As a part of our interest in recognition and cleavage of RNA we carried out thermal melting studies with the aim of screening a number of simple oligonucleotide modifications for their potential as modifying elements for RNA bulge stabilizing oligonucleotides. A specific model system from our studies on oligonucleotide-based artificial nuclease (OBAN) systems was chosen and the bulge size was varied from one to five nucleotides. Introduction of single 2'-modified nucleoside moieties (2'-O-methyl, 2'-deoxy and 2'-deoxy-2'-amino) with different conformational preferences adjacent to the bulge revealed that a higher preference for the north conformers gave more stable bulges across the whole range of bulge sizes. Changing a bulge closing a G-U wobble base pair to a G-C pair resulted in the interesting observation that, although the fully complementary complex and small bulges were highly stabilized, there was little difference in the stability of the larger bulges. The wobble base pair even gave a slight stabilization of the 5 nt bulge system. Introduction of a uridine C-5 linker with a single ammonium group was clearly bulge stabilizing (DeltaT(m) + 4.6 to + 5.4 degrees C for the three most stabilized bulges), although with limited selectivity for different bulge sizes since the fully complementary duplex was also stabilized. Introduction of a naphthoyl group on a 2'-aminolinker mostly gave a destabilizing effect, while introduction of a 5-aminoneocuproine moiety on the same linker resulted in stabilization of all bulges, in particular those with two or four unpaired nucleotides (DeltaT(m) + 3.6 and + 2.9 degrees C respectively). The aromatic groups destabilize the fully complementary duplex, resulting in higher selectivity towards stabilization of bulges. A combination of the studied partial element should be suitable for future designs of modified oligonucleotides that, apart from standard base pairing, can also provide additional non-Watson-Crick recognition of RNA. PMID- 15756641 TI - beta2-Adrenoceptor-mediated suppression of human intestinal mast cell functions is caused by disruption of filamentous actin dynamics. AB - Previous studies indicated potent inhibitory effects of beta2-adrenoceptor (beta2AR) activation on the immunological mediator release of mast cells (MC). Here, we studied effects of beta2AR agonists on human MC mediator release, and in particular on MC proliferation, adhesion, and migration. MC were isolated from human intestinal mucosa, purified, and cultured in the presence of stem cell factor (SCF). beta2AR activation by epinephrine, norepinephrine, and salbutamol suppressed the IgE receptor-dependent release of histamine, lipid mediators, and TNF-alpha, and inhibited SCF-dependent MC proliferation and migration. Moreover, beta2-adrenergic stimulation interfered with MC adhesion to fibronectin and human endothelial cells. Using fluorescent phallacidin, we found that beta2AR activation reduced the amount of filamentous actin (F-actin) within minutes, whereas MC stimulation by either IgE receptor cross-linking or SCF caused F-actin accumulation. Interestingly, this activation-induced F-actin increase was abolished by previous beta2-adrenergic stimulation. Finally, we demonstrated that disruption of the F-actin cytoskeleton by latrunculin B mimicked the effects of beta2AR agonists on MC adhesion and migration. Our results argue for an important role of F-actin interference in beta2AR-mediated MC inhibition. Furthermore, the data support the concept of neuroimmune interactions regulating intestinal MC distribution, density, and functionality in vivo. PMID- 15756642 TI - Kinetics of costimulatory molecule expression by T cells and dendritic cells during the induction of tolerance versus immunity in vivo. AB - Steady-state dendritic cells (DC) present peptide-MHC complexes to T cells in a tolerogenic manner, presumably because of deficient costimulation. However, it is clear that the path to tolerance involves initial T cell activation, suggesting that the deficit may lie in late-acting costimulatory molecules. With this in mind we have investigated the kinetics of expression of several costimulatory pairs on DC and OVA-reactive T cells after i.v. injection of mice with peptide and LPS (immunity), or peptide alone (tolerance). We find that T cells up regulate CD154, OX40, RANKL and PD-1 whether they are destined for tolerance or immunity, although there are some differences in the levels and length of expression. In contrast, when analyzing DC, we found that up-regulation of CD80, CD86, CD40, RANK and PDL-1 occurred only when peptide was co-administered with LPS. These data give a picture of the T cell looking for costimulatory cues that are not forthcoming when pMHC is presented by steady-state DC, leading to tolerance. However, we did see a strong and rapid up-regulation of RANKL on T cells that occurred specifically when peptide was given in the absence of LPS, suggesting a possible positive signal influencing the decision between tolerance and immunity. PMID- 15756643 TI - A polyclonal anti-vaccine CD4 T cell response detected with HLA-DP4 multimers in a melanoma patient vaccinated with MAGE-3.DP4-peptide-pulsed dendritic cells. AB - During the last few years, HLA class I tetramers have been successfully used to demonstrate anti-vaccine CD8 CTL proliferation in cancer patients vaccinated with tumor antigens. Frequencies of CTL as low as 10(-6) among CD8 cells were observed even in patients showing tumor regression. Little is known about the role of tumor-antigen-specific CD4 T cells in the context of these anti-vaccine responses. Therefore, we developed a very sensitive approach using fluorescent class-II-peptide multimers to detect antigen-specific CD4 T cells in vaccinated cancer patients. We produced HLA-DP4 multimers loaded with the MAGE-3(243-258) peptide and used them to stain ex vivo PBL from melanoma patients injected with dendritic cells pulsed with several class I and class II tumor antigenic peptides, including the MAGE-3(243-258) peptide. The multimer(+) CD4 T cells were sorted and amplified in clonal conditions; specificity was assessed by their ability to secrete IFN-gamma upon contact with the MAGE-3 antigen. We detected frequencies of about 1x10(-6) anti-MAGE-3.DP4 cells among CD4 cells. A detailed analysis of one patient showed an anti-MAGE-3.DP4 CD4 T cell amplification of at least 3000-fold upon immunization. TCR analysis of the clones from this patient demonstrated a polyclonal response against the MAGE-3 peptide. PMID- 15756644 TI - The advantage of mucosal immunization for polysaccharide-specific memory responses in early life. AB - The aim of vaccination is to rapidly elicit protective immunity and generate memory for sustained protection. We studied the induction and persistence of polysaccharide (PS)-specific memory in neonatal and infant mice primed with pneumococcal conjugate (Pnc1-TT) by assessing the response to native pneumococcal PS (PPS-1), the kinetics of the PPS-1-specific IgG response to a second Pnc1-TT dose and affinity maturation. A subcutaneous (s.c.) Pnc1-TT booster induced a rapid increase in PPS-1-specific IgG, indicating efficient priming for memory by a single dose of Pnc1-TT already at 1 week of age. High levels were maintained for >12 weeks. However, a PPS-1 booster induced no response in neonatal or infant mice. The adjuvant LT-K63 significantly enhanced the IgG response and affinity to Pnc1-TT by both the s.c. and the intranasal (i.n.) route in all age groups. In neonatal and infant mice, PPS-1 and LT-K63 induced a booster response only when given i.n. following either s.c. or i.n. priming with Pnc1-TT and LT-K63. In contrast, PPS-1 with or without LT-K63 administered s.c. compromised the ongoing PPS-1-specific response elicited in neonatal mice by either s.c. or i.n. priming with Pnc1-TT and LT-K63. These results demonstrate the advantage of the mucosal route for elicitation of PS-specific memory responses in early life. PMID- 15756645 TI - Evolution of diverse antiviral CD8+ T cell populations after murine cytomegalovirus infection. AB - Cytomegalovirus (CMV) is a major human pathogen normally controlled by cellular immune responses. The infection can be modeled in the mouse using murine CMV (MCMV). During the latent phase of infection, two different patterns of CD8(+) T cell responses have been observed: some specificities show increasing frequencies over time ("memory inflation"), while others, which are present acutely, are barely detectable at later time points. This distinction is independent of initial immunodominance. We analyzed the extent to which such responses differ functionally and tracked both their population distribution and their evolution over time. We observed two clear patterns of memory development that diverged early after infection. Acutely, CD8(+) T cells directed against all epitopes showed similar activation, phenotype and distribution. Thereafter, one set of responses ("inflationary") increased in frequency over time, was found in high numbers in non-lymphoid organs and was associated with an activated (CD28(low) CD27(low)CD122(low)) phenotype. In contrast, CD8(+) T cells responses specific for other MCMV epitopes ("non-inflationary") showed a slow reversion to a classical "central" memory phenotype without enrichment in non-lymphoid organs. A simple model to describe the equilibrium state in MCMV is presented, which may point to previously unexplored antiviral populations present after human CMV infection. PMID- 15756646 TI - B cells can prime naive CD4+ T cells in vivo in the absence of other professional antigen-presenting cells in a CD154-CD40-dependent manner. AB - The role of B cells as APC is well established. However, their ability to prime naive T cells in vivo has been difficult to examine because of the presence of dendritic cells. The current studies were undertaken to examine this issue in a model of adoptive transfer of antigen-specific B cells and T cells into histoincompatible Rag2(-/-) mice. By means of this system, we were able to demonstrate that antigen-specific B cells are competent APC for naive CD4(+) T cells specific for the same antigen. In vivo antigen presentation resulted in expansion of both CD4(+) T cells and B cells. The antigen-presenting function of the transferred B cells was dependent on the CD154-CD40 interaction, as transfer of CD154-deficient antigen-specific CD4(+) T cells or CD40-deficient B cells failed to induce T and B cell expansion in response to immunization. These results indicate that antigen-specific B cells have the capacity to induce primary T cell responses in the absence of other competent APC. PMID- 15756648 TI - The rat orthologue to the inhibitory receptor gp49B is expressed by neutrophils and monocytes, but not by NK cells or mast cells. AB - Mouse gp49B is a member of the leukocyte immunoglobulin-like receptor family. It is constitutively expressed by mast cells and certain myeloid cells, and expression can be induced on natural killer (NK) cells and T cells. We have cloned several rat cDNA, 78% identical to mouse gp49B at the amino acid level, that represent the rat orthologue to mouse gp49B. A mouse monoclonal antibody (WEN29) against rat gp49B was generated. By flow cytometry and Northern blot analysis, gp49B was found to be expressed by neutrophils and monocytes, but not NK cells (primary or IL-2-activated), T cells (resting or concanavalin A stimulated) or peritoneal mast cells. Following pervanadate treatment, the tyrosine phosphatase SHP-1 was co-immunoprecipitated with gp49B in the macrophage cell line R2. In glutathione S-transferase pull-down experiments, the cytoplasmic tail of rat gp49B associated with the SH2 domains of both SHP-1 and SHP-2, dependent on intact and phosphorylated immunoreceptor tyrosine-based inhibition motifs (ITIM). Compared to mouse, the cytoplasmic domain of rat gp49B contains a third ITIM-like sequence (YLYASV) that was phosphorylated by several Src family tyrosine kinases, enhanced the phosphorylation of other ITIM, and bound to the SH2 domains of SHP-2, suggesting a role in the recruitment of downstream phosphatases. PMID- 15756649 TI - Stable inhibition of hepatitis B virus proteins by small interfering RNA expressed from viral vectors. AB - BACKGROUND: There has been much research into the use of RNA interference (RNAi) for the treatment of human diseases. Many viruses, including hepatitis B virus (HBV), are susceptible to inhibition by this mechanism. However, for RNAi to be effective therapeutically, a suitable delivery system is required. METHODS: Here we identify an RNAi sequence active against the HBV surface antigen (HBsAg), and demonstrate its expression from a polymerase III expression cassette. The expression cassette was inserted into two different vector systems, based on either prototype foamy virus (PFV) or adeno-associated virus (AAV), both of which are non-pathogenic and capable of integration into cellular DNA. The vectors containing the HBV-targeted RNAi molecule were introduced into 293T.HBs cells, a cell line stably expressing HBsAg. The vectors were also assessed in HepG2.2.15 cells, which secrete infectious HBV virions. RESULTS: Seven days post transduction, a knockdown of HBsAg by approximately 90%, compared with controls, was detected in 293T.HBs cells transduced by shRNA encoding PFV and AAV vectors. This reduction has been observed up to 5 months post-transduction in single cell clones. Both vectors successfully inhibited HBsAg expression from HepG2.2.15 cells even in the presence of HBV replication. RT-PCR of RNA extracted from these cells showed a reduction in the level of HBV pre-genomic RNA, an essential replication intermediate and messenger RNA for HBV core and polymerase proteins, as well as the HBsAg messenger RNA. CONCLUSIONS: This work is the first to demonstrate that delivery of RNAi by viral vectors has therapeutic potential for chronic HBV infection and establishes the ground work for the use of such vectors in vivo. PMID- 15756650 TI - Improved neuronal transgene expression from an AAV-2 vector with a hybrid CMV enhancer/PDGF-beta promoter. AB - BACKGROUND: Adeno-associated virus type 2 (AAV-2) vectors are highly promising tools for gene therapy of neurological disorders. After accommodating a cellular promoter, AAV-2 vectors are able to drive sustained expression of transgene in the brain. This study aimed to develop AAV-2 vectors that also facilitate a high level of neuronal expression by enhancing the strength of a neuron-specific promoter, the human platelet-derived growth factor beta-chain (PDGF) promoter. METHODS AND RESULTS: A hybrid promoter approach was adopted to fuse the enhancer of human cytomegalovirus immediately early (CMV) promoter to the PDGF promoter. In cultured cortex neurons, AAV-2 vectors containing the hybrid promoter augmented transgene expression up to 20-fold over that mediated by titer-matched AAV-2 vectors with the PDGF promoter alone and 4-fold over the CMV enhancer/promoter. Injection of AAV-2 vectors with the hybrid promoter into the rat striatum resulted in neuron-specific transgene expression, the level of which was about 10-fold higher than those provided by the two control AAV-2 expression cassettes at 4 weeks post-injection and maintained for at least 12 weeks. Gene expression in the substantia nigra through possible retrograde transport of the AAV-2 vectors injected into the striatum was not obvious. After direct injection of AAV-2 vectors into the substantia nigra, transgene expression driven by the hybrid promoter was observed specifically in dopaminergic neurons and its level was about 3 and 17 times higher than that provided by the PDGF promoter alone and the CMV enhancer/promoter, respectively. CONCLUSIONS: Enhanced transgene capacity plus neuron-specificity of the AAV-2 vectors developed in this study might prove valuable for gene therapy of Parkinson's disease. PMID- 15756651 TI - Shakespeare on parkinsonism. PMID- 15756652 TI - Levetiracetam for the treatment of essential tremor. PMID- 15756653 TI - Differences in tumor core distribution between palpable and nonpalpable prostate tumors in patients diagnosed using extensive transperineal ultrasound-guided template prostate biopsy. AB - BACKGROUND: The authors performed extensive transperineal ultrasound-guided template prostate biopsies to investigate carcinoma core distribution. METHODS: Between August 2000 and May 2004, 371 men underwent template biopsies. Three hundred twelve patients had not undergone a previous biopsy (first group) and 59 had undergone previous transrectal sextant biopsies (repeat group). Of the 312 patients in the first group, 236 had normal digital rectal examination (DRE) findings (DRE- first group) and 76 patients had an abnormal DRE (DRE+ first group). A mean of 20.1 biopsy cores (range, 9-38 cores) was taken from the entire prostate. The region > 2.0 cm from the rectal face of the prostate was defined as the anterior region and the remaining area was defined as the posterior region. RESULTS: In the DRE- first group, the carcinoma core rate (number of tumor cores/number of biopsy cores) in the anterior region (7.2%) did not differ from that of the posterior region (7.3%) (P = 0.9635). However, in the DRE+ first group, the carcinoma core rate in the posterior region (22.0%) was found to be higher than in the anterior region (13.2%) (P < 0.0001). In the repeat group, the carcinoma core rate in the posterior region (3.1%) was significantly (P = 0.0008) lower than that exhibited in the anterior region (7.2%). CONCLUSIONS: The results of the current study suggest that nonpalpable prostate carcinoma is distributed equally within the entire prostate, although palpable carcinoma is distributed mainly in the posterior region and many of the tumor foci in the anterior region may be missed by a transrectal sextant biopsy. The examination of radical prostatectomy specimens is required to prove these results. PMID- 15756654 TI - Intrathecal baclofen in the treatment of painful, disabling spasms in Friedreich's ataxia. AB - We report on the case of a patient suffering from Friedreich's ataxia (FA) with very painful and disabling spasms that were improved markedly by intrathecal baclofen infusion. To our knowledge, this is the first report of an intrathecal baclofen-delivering pump implantation in an FA patient. PMID- 15756655 TI - A longitudinal study of palliative care: patient-evaluated outcome and impact of attrition. AB - BACKGROUND: The current article evaluated the course of patient-assessed symptomatology in specialized palliative care and tested for bias due to patient attrition in measures of initial symptomatology and treatment outcome. METHODS: Over 2 years, 267 consecutive, eligible patients were referred to a department of palliative care. Upon arrival, 201 patients consented to participate in a questionnaire-based evaluation of quality of life (QOL). Of these, 175 patients participated, and 142, 119, and 95 participated in the study at 1, 2, and 3 weeks, respectively. Weekly, participants completed the self-assessment questionnaires European Organization for Research and Treatment of Cancer QLQ C30, Edmonton Symptom Assessment System, Hospital Anxiety and Depression Scale, and Multidimensional Fatigue Inventory. Physicians used the Mini Mental State Examination to evaluate cognitive function. Changes from the initial symptom scores for each week were calculated. Initial scoring and change after 1 week were tested for association with completion level, i.e., whether the patient completed questions at 1, 2, 3, or 4 time points. RESULTS: High initial symptom intensity and significant improvements over time were observed for pain, lack of appetite, nausea/vomiting, sleeplessness, constipation, and overall QOL/well being. For some symptoms, initial scores were significantly higher in patients who dropped out, but the changes over the first week were not significantly different between completion levels for any symptom. CONCLUSIONS: Improvement in symptom intensity was identified. Dropout was associated with higher initial symptomatology but not with poorer outcome of palliative treatment. PMID- 15756656 TI - Postprostatectomy adjuvant versus salvage radiotherapy: a complication-adjusted number-needed-to-treat analysis. AB - BACKGROUND: Radiotherapy (RT) has been used with success after radical retropubic prostatectomy (RRP), both in the adjuvant and salvage settings. The purpose of the current investigation was to systematically compare adjuvant versus salvage RT in a manner that incorporates both treatment efficacy and complications. METHODS: A literature review was performed of reports of post-RRP salvage and adjuvant RT, and 12 trials comprising 1060 patients met the appropriate inclusion criteria. The biochemical failure-free survival in each study/arm was tabulated, and these values were entered into a model to compute an unadjusted number-needed to treat (NNT). RT complications were then considered, accounting for differences in toxicity incidences in the salvage versus adjuvant setting, to compute complication-adjusted NNTs. In all the trials, the signs and magnitudes of the NNTs obtained were used to compare adjuvant with salvage RT. RESULTS: The absolute NNT analysis showed an advantage of adjuvant compared with salvage RT. After adjustment for RT complications, however, the advantage shifted to salvage RT. This transition point from superiority of adjuvant RT to superiority of salvage RT was sensitive to the estimated incidence and severity of RT side effects. CONCLUSIONS: Adjuvant post-RRP RT was advantageous in comparison to salvage RT if the side effects of RT were estimated to be negligible. However, with moderate incidence/severity of RT side effects, salvage RT was advantageous. The findings herein must be tested in a prospective study in which both health related quality of life and cancer control are documented in patients receiving adjuvant versus salvage post-RRP RT. Further work is needed to better estimate parameters entered into the model to determine the precise transition point between adjuvant and salvage RT with modern RT techniques. PMID- 15756657 TI - Biostability of polyether-urethane scaffolds: a comparison of two novel processing methods and the effect of higher gamma-irradiation dose. AB - This article deals with enzyme-induced biodegradation behavior of thermoplastic polyether-urethane (TPU). Porous scaffolds were processed by a new foaming method applied in hot pressing and injection molding. The scaffolds were subsequently gamma sterilized. The samples were incubated with cholesterol esterase (CE) for 28 days to simulate an enzymatic degradation order to assess polymer biostability. The main focus of degradation products was the most toxic one: methylene dianiline (MDA). LC/MS was used to separate the breakdown products and to identify possible MDA amounts. The results showed that (a) the hot-pressed sample released an MDA amount almost twice as large (0.26 ng +/- 0.008) as that of the injection-molded samples (0.15 ng +/- 0.003) after incubation with enzyme activity in the physiological range, and (b) a tenfold increase in CE activity revealed considerably higher MDA amounts (7540.0 ng +/- 0.004). This enzyme concentration is physiologically unlikely, however, but may occur for extreme high inflammation behavior. Even for extremely high levels of CE enzyme, the scaffold will not discharge MDA above toxic levels. The injection-molded samples sterilized at 25 kGy seem to represent the most promising processing method. Therefore, the new injection-molding foaming process of polyether-urethane can be considered appropriate for use as a biomaterial. PMID- 15756658 TI - Cost utility in the United States of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone for the treatment of elderly patients with diffuse large B-cell lymphoma. AB - BACKGROUND: Findings from the Groupe d'Etude des Lymphomes Adultes LNH 98-5 study showed that rituximab added to combined cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) prolonged progression-free survival and overall survival in adults age >/= 60 years with diffuse large B-cell non-Hodgkin lymphoma (DLBCL). The current study was conducted to investigate the incremental cost utility of the addition of rituximab to CHOP (R-CHOP) compared with CHOP alone. METHODS: Clinical prognosis of the time to disease progression and death was estimated using published evidence from the LNH 98-5 study (n = 399 patients) that was linked mathematically to published long-term outcome data on patients with DLBCL. Drug-acquisition costs were based on published data from formulary pricing sources, and the costs of cancer surveillance and end-of-life care were based on published literature sources. The authors assessed cost utility as the difference in costs between R-CHOP and CHOP divided by the increase in expected overall survival adjusted for quality of life. RESULTS: Over 5 years, it was projected that R-CHOP would prolong overall survival by 1.04 years. The mean cumulative cost of CHOP was 3358 dollars, and the mean cost of R-CHOP was 17,225 dollars, resulting in a cumulative net increase of 13,867 dollars. The posttreatment cancer surveillance cost for CHOP was 3950 dollars, compared with 5202 dollars for R-CHOP. It was estimated that R-CHOP would have a cost-utility ratio of 19,297 dollars per year of life gained compared with CHOP when adjusted for quality of life. R-CHOP remained cost effective over wide ranges of variables in sensitivity analyses. CONCLUSIONS: Compared with CHOP alone, it was predicted that R-CHOP would be cost effective in elderly patients with DLBCL. PMID- 15756659 TI - Sonographic features of Ebstein anomaly associated with hydrops fetalis: a report of two cases. AB - Ebstein anomaly, an abnormally low insertion of the tricuspid valve, occurs in 0.5% of patients with congenital heart disease. In rare cases, this disorder may be complicated by congestive heart failure in utero and hydrops fetalis. This article reports the prenatal sonographic features of 2 cases of Ebstein anomaly associated with hydrops fetalis. In both cases, fetal echocardiography was performed at 34 weeks of gestation. The 4-chamber view showed fetal cardiomegaly and pericardial effusion. In both cases, the annular attachment of the tricuspid valve leaflets was difficult to demonstrate and so distal that it could easily be confused with papillary muscle in the right ventricular wall. Poor fetal cardiac hemodynamics with severe tricuspid regurgitation was demonstrated by pulsed Doppler imaging. Pulmonary regurgitation was also clearly demonstrated in both cases. Preterm delivery with stillbirth occurred in both cases. Autopsies confirmed the very distal displacement of the tricuspid valve insertion, close to the apex, and enlargement of the right atrium. PMID- 15756660 TI - Extranodal primary B-cell non-Hodgkin lymphoma of the breast mimicking acute mastitis. AB - We report a case of primary, high-grade non-Hodgkin B-cell lymphoma in the breast of a young woman. The clinical and sonographic presentation was not of a mass but of an infiltrating anechoic process mimicking mastitis. Primary breast lymphoma is a rare entity, especially in young females. Early detection and treatment are crucial because the results of salvage treatment are generally poor. In previous imaging reports of breast lymphoma, it has always been considered as a mass, though the presence of markedly hypoechoic regions that look like fluid collections is a well known sonographic characteristic of lymphoma. PMID- 15756661 TI - Doppler sonography of the normal lacrimal gland. AB - PURPOSE: We used Doppler sonography to determine the resistance index (RI) and pulsatility index (PI) of the normal lacrimal artery (LA) in both females and males. We also compared the values obtained at various periods of reproductive life. METHODS: The study was performed in 25 prepubertal girls, 28 females of reproductive age, and 27 postmenopausal women, 23 pregnant women, and 104 healthy males. Doppler sonography was used to determine the RI and PI of the LA. RESULTS: The mean PI for the entire patient population was 1.48 +/- 0.60 and the RI was 0.72 +/- 0.09. The RI and PI values of the LA did not differ between males and females. Similarly, the mean RI and PI values of the LA did not change significantly between the various reproductive phases. CONCLUSION: The PI and RI of the lacrimal gland are similar in both sexes. Moreover, they are not altered by changes in levels of sex steroids. PMID- 15756662 TI - Pericentric inversion of chromosome 2 and echogenic vasculature in the basal ganglia: a new finding? AB - Linear branching echogenicities in the thalamus or basal ganglia have been reported in infants with several genetic and nongenetic disorders. In this article, we report 2 cases of newborns with a neurosonographic diagnosis of thalamic/basal ganglia vasculopathy and karyotype analysis showing pericentric inversion of chromosome 2. To our knowledge, there has been no previous mention of an association between these entities. PMID- 15756664 TI - Correlation between fetal gastric size and amniotic fluid volume. AB - PURPOSE: Since abnormal conditions of the fetal digestive tract may alter both amniotic fluid volume and fetal gastric volume, we sought to determine whether amniotic fluid volume is correlated with fetal gastric volume in normal pregnancy. METHODS: A total of 280 fetal gastric size measurements were made prospectively from routine sonographic examinations of women with normal singleton pregnancies between 16 and 42 weeks of gestation. The fetal stomach was defined as the largest area including the pyloric site on transverse or oblique real-time sonographic scans. Gastric volume was calculated according to the formula for a prolate ellipsoid. The amniotic fluid index (AFI) was used for the evaluation of amniotic fluid volume. RESULTS: Both fetal gastric volume and AFI were significantly correlated with gestational age (R2= 0.422 and R2= 0.128, respectively). Only a weak correlation was found between gastric volume and AFI (R2= 0.036, p <0.001). On multivariate linear regression analysis adjusting for gestational age and fetal biometric measurements, gastric volume was not an independent and significant predictor of AFI. CONCLUSIONS: Although sonographically determined fetal gastric volume measurements appear to be useful in the assessment of fetal digestive tract anomalies, fetal gastric volume has no clinically significant effect on the amniotic fluid volume in normal pregnancy. PMID- 15756663 TI - Three-dimensional reconstruction of endosonographic images of gastric lesions: preliminary experience. AB - PURPOSE: This study was conducted to assess the ability of the endoluminal sonographic probes to image upper gastrointestinal lesions detected with conventional endoscopy and to reconstruct 3-dimensional (3D) images. METHODS: Sonographic images were obtained from 25 gastrointestinal lesions in 22 patients using a sonographic miniprobe manually drawn at a set speed across lesions detected during conventional endoscopy. 3D images were then reconstructed using an in-house software program. RESULTS: In 16 lesions (64%), the 3D sectional images were evaluated as good, and all 25 lesions (100%) could be visualized both transversely and longitudinally. In 12 lesions (48%), the 3D display of the endosonographic surface image corresponded to the endoscopic appearance. In all 11 cases of gastric cancer, the depth of tumor invasion measured using the 3D image was consistent with the histologic diagnosis (100% accuracy). When videotaped sonographic images were stored on a hard disk, it took an average of 3.5 minutes to obtain and display 3D images of the targeted area. CONCLUSIONS: 3D sonographic reconstructions of digestive lesions were useful in diagnosis, and we believe that further improvements should enhance the clinical usefulness of 3D endosonography. PMID- 15756665 TI - Ultrastructural biologic effects of sonography with pulse inversion and microbubble contrast in rabbit liver. AB - PURPOSE: This prospective study was conducted to evaluate the biologic effects of microbubble destruction with pulse-inversion harmonic imaging on rabbit liver parenchyma. METHODS: The livers of 6 albino rabbits were examined sonographically by a single investigator. Three rabbits underwent contrast-enhanced sonography, with scanning starting 5 seconds after injection by using pulse-inversion harmonic imaging with a mechanical index of 1.2. Four time-triggered images were recorded at a rate of 1 frame every 2 seconds. For comparison, 3 control rabbits had pulse-inversion harmonic imaging with a mechanical index of 1.2 only, without contrast medium. Immediately after sonography, the animals were killed and uninterrupted, thin serial sections of the liver from both groups were analyzed by energy-filtered transmission electron microscopy. RESULTS: The hepatic parenchyma of rabbits exposed to contrast agents had ultrastructural damage: mitochondria with fragmented crests; interrupted rough endoplasmic reticulum; enlarged intercellular spaces; highly vacuolized cytoplasmic areas; dilated sinusoids, sometimes with an irregular and interrupted endothelial wall; fragmented hepatocyte microvilli in dilated spaces of Disse; fragmented or missing microvilli in bile canaliculi; vacuolated and lysosome-deprived hepatic cytoplasm around the bile canaliculi; markedly injured or fragmented endothelium in larger vessels; and damaged basal membrane. Control-group results indicated that exposure to ultrasound alone did not cause ultrastructural damage to hepatic cells. CONCLUSIONS: Simultaneous exposure to contrast administration and pulse inversion harmonic imaging with a high mechanical index causes ultrastructural damage in the rabbit liver. PMID- 15756666 TI - Vascular and nonvascular complications of renal transplants: sonographic evaluation and correlation with other imaging modalities, surgery, and pathology. AB - Cadaveric or living donor renal transplantation is commonly performed in individuals with end-stage renal disease. In recent years, gray-scale sonography, coupled with color Doppler sonography (CDUS), power Doppler sonography (PDUS), or spectral Doppler sonography, has become the primary imaging modality for these patients. Postoperative serial sonography is performed to detect complications and aid in posttransplant management. In addition, sonography is used to guide percutaneous aspiration of fluid or biopsy to diagnose rejection or renal and perirenal masses. In this article we discuss the spectrum of sonographic findings, both vascular and nonvascular, of renal transplant complications, including but not limited to renal arterial and venous stenosis and thrombosis, peritransplant collections (lymphoceles, hematomas, urinomas, and seromas), posttransplant lymphoproliferative disorder, and postbiopsy complications (hematomas, pseudoaneurysms, and arteriovenous fistulas). We correlate sonographic findings with those from other imaging modalities (such as angiography, CT, and MRI) and findings at surgery and pathology when possible. PMID- 15756667 TI - Effect of diffuse fatty infiltration of the liver on hepatic artery resistance index. AB - PURPOSE: This study was conducted to evaluate the effect of various degrees of diffuse fatty infiltration of the liver on the hepatic artery resistance index. METHODS: One-hundred forty subjects were examined using standard color and spectral Doppler sonography protocols. Fatty infiltration of the liver was identified and graded sonographically. The patients were grouped (n = 35 in each of 4 groups) according to the degree of diffuse fatty infiltration of the liver as follows: normal (group 1), mild (group 2), moderate (group 3), and severe (group 4). The resistance index calculated for each patient was the mean of 3 measurements. Mean resistance index of the hepatic artery was then calculated for each group. RESULTS: The mean resistance index was 0.81 +/- 0.04 for group 1, 0.79 +/- 0.06 for group 2, 0.75 +/- 0.05 for group 3, and 0.73 +/- 0.05 for group 4. We found a statistically significant (p < 0.05) decrease in resistance index when comparing groups 3 and 4 with groups 1 and 2 separately. CONCLUSIONS: Hepatic artery resistance index decreases as the severity of diffuse fatty infiltration increases. PMID- 15756668 TI - Differential diagnosis of hepatic cystic lesions with gray-scale and color Doppler sonography. AB - PURPOSE: This study was conducted to evaluate the effectiveness of sonography, especially color Doppler sonography, in the differential diagnosis of cystic hepatic lesions. METHODS: Ninety-two pathologically or clinically proven hepatic cystic lesions (20 cystic malignancies, 24 abscesses, and 48 simple cysts) were evaluated with gray-scale and color Doppler sonography. The sonographic features were analyzed retrospectively. The percentage, sensitivity, specificity, and positive and negative predictive values of the sonographic features of each disease category were calculated. RESULTS: On gray-scale sonography, the simple cysts were easily distinguished from cystic malignancies and abscesses. While no significant differences were found between hepatic cystic malignancies and hepatic abscesses with respect to the number, shape, margin status, and presence of thick wall of the lesion, the presence of septation and mural nodules was significantly higher in the cystic malignancies than in abscesses. The sensitivity and specificity of color Doppler sonography in differentiating cystic malignancies from abscesses and simple cysts were 85% and 96%, respectively. CONCLUSIONS: Color Doppler sonography provides information about blood flow that supplements that gained on gray-scale sonography, and the presence of color signals in the solid portion of the cystic lesions carries a high diagnostic value in differentiating hepatic cystic malignancies from abscesses and simple cysts. PMID- 15756670 TI - Rare right ventricular myxoma detected incidentally on a FAST examination. AB - A young man underwent Focused Assessment with Sonography in Trauma (FAST) scan for blunt abdominal trauma. An incidental finding on the pericardial view was the presence of a tumor in the right ventricle. Detailed cardiac imaging and subsequent excision revealed the presence of a myxoma. The patient had no obvious cardiac symptoms, but the awareness and experience of the examiner during the FAST examination detected this rare heart tumor at a latent stage, preventing serious, irreversible complications. This case demonstrates the potential importance of looking above the diaphragm during routine abdominal sonography, especially in patients who are critically ill. PMID- 15756672 TI - Ancient DNA analysis of human neolithic remains found in northeastern Siberia. AB - We successfully extracted DNA from a bone sample of a Neolithic skeleton (dated 3,600 +/- 60 years BP) excavated in northeastern Yakutia (east Siberia). Ancient DNA was analyzed by autosomal STRs (short tandem repeats) and by sequencing of the hypervariable region I (HV1) of the mitochondrial DNA (mtDNA) control region. The STR profile, the mitochondrial haplotype, and the haplogroup determined were compared with those of modern Eurasian and Native American populations. The results showed the affinity of this ancient skeleton with both east Siberian/Asian and Native American populations. PMID- 15756673 TI - Chlamydia trachomatis infection and persistence of human papillomavirus. AB - Human papillomavirus (HPV) persistence is the major cause of cervical cancer, but most HPV infections will not persist and risk factors for HPV persistence are not well known. Chlamydia (C.) trachomatis infection seems to also be associated with cervical cancer. We investigated whether C. trachomatis infection is a risk factor for HPV persistence. In a cohort of 12,527 women participating in a population-based HPV screening trial in Sweden, 6,418 women completed testing for HPV DNA by general primer PCR and typing by reverse dot blot hybridization. On average 19 months later, 303 women that had been HPV-positive and had normal cytology at enrollment completed a new HPV test. Environmental exposures were assessed by an 87-item questionnaire. Previous sexually transmitted infections were also investigated by serology. At follow-up, 44% of the women were positive for the same type of HPV DNA as at enrollment. Persistence correlated with length of follow-up (p < 0.01) and condom use seemed to protect against HPV persistence (p < 0.05). The most significant risk factor for persistent presence of HPV DNA was self-reported history of previous C. trachomatis infection (relative risk in multivariate model = 2.09; 95% confidence interval = 1.05-4.18). We conclude that persistence of oncogenic HPV infections is more likely among women with a previous C. trachomatis infection. PMID- 15756674 TI - Peripheral blood IFN-gamma-secreting Valpha24+Vbeta11+ NKT cell numbers are decreased in cancer patients independent of tumor type or tumor load. AB - Natural killer T (NKT) cells are CD1d-restricted lymphoid cells and are characterized by an invariant T-cell receptor, which in humans consists of a Valpha24 chain paired with a Vbeta11 chain. These cells are known for their rapid production of large amounts of cytokines (e.g., IFN-gamma and IL-4), thereby modulating other cells of the immune system such as T cells, NK cells and dendritic cells. NKT cells have been reported to play important regulatory roles in many immune responses, including antitumor immune responses. Here, we demonstrate an age-dependent decrease in circulating Valpha24(+)Vbeta11(+) NKT cell numbers in both healthy controls and cancer patients and demonstrate that in both groups females have higher NKT cell levels compared to males. In a large group of 120 cancer patients, we show that circulating Valpha24(+)Vbeta11(+) NKT cell numbers are about 50% lower than in age- and gender-matched healthy controls and that this decrease is independent of tumor type or tumor load. This decrease was not restored upon tumor removal by means of surgery or radiotherapy. Even though the percentage of NKT cells that secrete IFN-gamma, as detected by ELISPOT, is normal in cancer patients, the absolute number of circulating IFN gamma-secreting NKT cells is reduced. Together, our results suggest that the reduced circulating Valpha24(+)Vbeta11(+) NKT cell numbers in cancer patients are not affected by tumor load, but might actually reflect a risk factor for tumor development, e.g., by hampering efficient tumor immunosurveillance. PMID- 15756676 TI - Tumor suppressor gene Runx3 sensitizes gastric cancer cells to chemotherapeutic drugs by downregulating Bcl-2, MDR-1 and MRP-1. AB - The Runx3 gene is a member of the runt domain family transcription factors, key regulators of development and differentiation in metazoan. Recently, Runx3 was identified as a tumor suppressor gene. Loss of Runx3 was found to be associated with genesis and progression of gastric cancer. In this study, we transfected the gastric cancer cell line SGC7901 with eukaryotic expression vector of Runx3. In vitro drug sensitivity assay suggested that SGC7901/Runx3 cells were more sensitive to various chemotherapeutic drugs. Blocking Runx3 expression in immortalized stomach mucosal cells (GES-1) or gastric cancer cells (SGC7901) by Runx3-specific small interfering RNA conferred the cells resistance to chemotherapeutic drugs. Flow cytometry examination suggested that expression of Runx3 in gastric cancer cells increased the intracellular accumulation and retention of adriamycin. Semiquantitative RT-PCR and Western blot suggested that Runx3 downregulated expression of Bcl-2, MDR-1 (P-gp) and MRP-1. Binding of Runx3 to promoter sequences of Bcl-2, MDR-1 and MRP-1 gene was detected by eletrophoretic mobility shift assay (EMSA) and supershift EMSA. We cloned the MDR 1 and MRP-1 gene promoters containing Runx binding sites and constructed the luciferase reporter vectors of these 2 promoters. Luciferase reporter assay suggested that Runx3 inhibited the promoter activity of the MDR-1 and MRP-1 promoter in SGC7901 cells. Taken together, our findings suggested that overexpression of Runx3 could sensitize gastric cancer cells to chemotherapeutic drugs by downregulating the Bcl-2, MDR-1 and MRP-1. PMID- 15756675 TI - Necrotic death but not irradiation abolishes costimulation of T-cell effector functions and survival by CD80-expressing tumor cells. AB - Tumor vaccination by the use of gene-modified cancer cells that provide costimulatory signals has been successfully applied in preclinical animal models and is currently evaluated in a variety of clinical settings. In previous work, we demonstrated the efficacy of B7.1/CD80 to promote tumor immunity in syngeneic murine models and to prevent deletion of activated T cells by activation-induced cell death (AICD). In clinical trials, tumor cell vaccines are generally inactivated to avoid transfer of live tumor cells, i.e., additional tumor burden. Previous data indicated, however, that inactivation of tumor cells by lethal ionizing irradiation abrogates tumor vaccination by CD80-expressing cells. Here, we compare living and irradiated allogeneic tumor cells regarding their capacity to induce T-cell effector functions and their propensity to interfere with T-cell deletion by apoptosis. Both lethally irradiated and nonirradiated tumor cells facilitated T-cell proliferation, tumor cell lysis, and interfered with T-cell AICD to a similar extent. In contrast, necrotic tumor cells failed to costimulate T-cell effector functions. Thus, irradiation does not seem to hamper tumor cell mediated costimulation of T-cell effector functions. In contrast, necrosis of gene-modified tumor cells abrogates costimulation of T cells by CD80-expressing cells. PMID- 15756677 TI - Cumulative 5-year diagnoses of CIN2, CIN3 or cervical cancer after concurrent high-risk HPV and cytology testing in a primary screening setting. AB - The aim of our study was to assess the cumulative 5-year diagnoses of CIN2, CIN3 or invasive cervical cancer (CIN2+) after concurrent screening by high-risk HPV test and Pap smear in a primary screening setting. Four thousand thirty-four women from Eastern Thuringia/Germany were recruited from 1996 to 1998 for baseline screening that included routine cytology, high-risk HPV testing by consensus primer PCR GP5+/6+ and routine colposcopy. After a median of 59 months 3,153 women participated in final screening using identical methods. Women with abnormal cytology or colposcopy or a positive high-risk HPV test at any time during the study period were recalled for expert colposcopy and histologic verification. CIN2+ was detected in 160 women resulting in a cumulative 5-year proportion of 4.4% (95% CI: 3.7-5.0%). Of 3,702 women who were high-risk HPV negative at baseline, 34 (1.1-95% CI: 0.7-1.4%) had either prevalent CIN2+ or developed CIN2+ within the observation period. HPV/cytology double negatives at baseline were at lowest risk for CIN2+ (1.0-95% CI: 0.7-1.4%) compared to screening positives (16.8-100% depending on combined test results). The 5-year negative predictive value in HPV-/Cyto- women was 99.0% (95% CI: 98.6-99.3%). This suggests that a prolongation of the screening intervals in this group is feasible. However, it should be noted that 1 woman developed a microinvasive carcinoma within the observation period. Moreover, 2 women with prevalent cancer were missed by both tests. The prognostic relevance of concurrent high-risk HPV/cytology screening needs to be verified further by randomized trials. PMID- 15756678 TI - Tobacco-specific nitrosamines in smokeless tobacco products marketed in India. AB - Smokeless tobacco products are a known cause of oral cancer in India. Carcinogenic tobacco-specific nitrosamines in these products are believed to be at least partially responsible for cancer induction, but there have been no recent analyses of their amounts. We quantified levels of 4 tobacco-specific nitrosamines, N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT), N' nitrosoanabasine (NAB) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), in 32 products marketed currently in India. Levels of nitrate, nitrite and nicotine were also determined. The highest levels of tobacco-specific nitrosamines were found in certain brands of khaini, zarda and other smokeless tobacco products. Concentrations of NNN and NNK in these products ranged from 1.74-76.9 and 0.08-28.4 microg/g, respectively. Levels of tobacco-specific nitrosamines in gutka were generally somewhat lower than in these products, but still considerably higher than nitrosamine levels in food. Tobacco-specific nitrosamines were rarely detected in supari, which does not contain tobacco, or in tooth powders. The results of our study demonstrate that exposure to substantial amounts of carcinogenic tobacco-specific nitrosamines through use of smokeless tobacco products remains a major problem in India. PMID- 15756679 TI - Dietary patterns and the risk of postmenopausal breast cancer. AB - The association between individual foods and breast cancer has been inconsistent. Therefore, we examined the association between diet and risk of postmenopausal breast cancer by the alternative approach of dietary patterns. Dietary patterns were identified with factor analysis from food consumption data collected from a food frequency questionnaire in 1984. Relative risks were computed using proportional hazard models and adjusted for known risk factors for breast cancer. Between 1984 and 2000, we ascertained 3,026 incident cases of postmenopausal breast cancer. We identified 2 major dietary patterns. The prudent pattern is characterized by higher intake of fruits, vegetables, whole grains, low-fat dairy products, fish and poultry, while the Western pattern is characterized by higher intake of red and processed meats, refined grains, sweets and desserts and high fat dairy products. Neither of the patterns was associated with overall risk of postmenopausal breast cancer. However, a positive association between the Western pattern score was observed among smokers at baseline (relative risk = 1.44, comparing top to bottom quintiles; 95% CI = 1.02-2.03; p for trend = 0.03). An inverse association was observed between the prudent pattern and estrogen receptor-negative cancer (relative risk = 0.62; 95% CI = 0.45-0.88; p for trend = 0.006). Among the major food groups, higher consumptions of fruits (relative risk for 1 serving/day increase = 0.88; 95% CI = 0.80-0.97; p = 0.009) and vegetables (relative risk = 0.94; 95% CI = 0.88-0.99; p = 0.03) were significantly associated with decreased risk for ER(-) breast cancer. In conclusion, we did not observe an overall association between the prudent or Western pattern and overall breast cancer risk. However, a Western-type diet may elevate risk of breast cancer among smokers, and a prudent diet may protect against estrogen receptive negative tumors. PMID- 15756680 TI - The role of vegetable and fruit consumption in the aetiology of squamous cell carcinoma of the oesophagus: a case-control study in Uruguay. AB - There is strong evidence of a protection of high consumption of plant foods against squamous cell carcinoma of the oesophagus, but the respective role of fruits and vegetables is not well elucidated. In the time period 1996-2003, a case-control study designed in order to address this question was conducted in Uruguay. The study included 200 cases and 400 age- and sex-matched controls. High total vegetable and total fruit consumption was the strongest protective food group for this malignancy (OR 0.31, 95% CI 0.18-0.51). When this food group was dissected in total vegetables and total fruits, the latter item was more protective than the former one (OR for total fruits 0.48, 95% CI 0.35-0.66) (p value for difference between both variables 0.04). Finally, when total fruits were dissected in citrus fruits and other fruits, citrus fruits were strongly associated with a reduction in risk (OR 0.47, 95% CI 0.34-0.67). The results of our study point towards an important role of fruits in the prevention of squamous cell carcinoma of the oesophagus. PMID- 15756681 TI - Clinicopathologic and prognostic values of apolipoprotein D alterations in hepatocellular carcinoma. AB - We previously identified apolipoprotein D (Apo D) as a novel tumor suppressor gene based on the pharmacological unmasking of epigenetic silencing. We analyzed Apo D expression using real-time reverse transcription-PCR and evaluated its expression status based on the clinicopathological parameters of 70 patients with hepatocellular carcinoma (HCC). Immunohistochemical staining was also performed. We determined the methylation status of Apo D gene promoter by methylation specific PCR (MSP). The Apo D gene-expression in tumor tissue was significantly lower than that in nontumor tissue (p = 0.011). A low Apo D expression significantly correlated with less-differentiated HCC (p = 0.019). Immunohistochemical studies confirmed a decreased Apo D expression in poorly differentiated tumors. The prognosis of patients with a lower Apo D gene expression was significantly worse than that in those with a higher expression (p = 0.028). The Apo D gene-expression was an independent prognostic factor (relative risk: 2.36, p = 0.018). An MSP assay showed a low-level of methylation in well differentiated HCC and a high-level of methylation in less differentiated tumors. Apo D may be a novel tumor suppressor gene of HCC, and its expression status may be a useful biomarker for predicting the patient outcome. PMID- 15756682 TI - Occupational risk factors for low grade and high grade glioma. PMID- 15756683 TI - Bystander signaling between glioma cells and fibroblasts targeted with counted particles. AB - Radiation-induced bystander effects may play an important role in cancer risks associated with environmental, occupational and medical exposures and they may also present a therapeutic opportunity to modulate the efficacy of radiotherapy. However, the mechanisms underpinning these responses between tumor and normal cells are poorly understood. Using a microbeam, we investigated interactions between T98G malignant glioma cells and AG01522 normal fibroblasts by targeting cells through their nuclei in one population, then detecting cellular responses in the other co-cultured non-irradiated population. It was found that when a fraction of cells was individually irradiated with exactly 1 or 5 helium particles ((3)He(2+)), the yield of micronuclei (MN) in the non-irradiated population was significantly increased. This increase was not related to the fraction of cells targeted or the number of particles delivered to those cells. Even when one cell was targeted with a single (3)He(2+), the induction of MN in the bystander non-irradiated population could be increased by 79% for AG01522 and 28% for T98G. Furthermore, studies showed that nitric oxide (NO) and reactive oxygen species (ROS) were involved in these bystander responses. Following nuclear irradiation in only 1% of cells, the NO level in the T98G population was increased by 31% and the ROS level in the AG0 population was increased by 18%. Treatment of cultures with 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1 oxyl-3-oxide (c-PTIO), an NO scavenger, abolished the bystander MN induction in non-irradiated AG01522 cells but only partially in non-irradiated T98G cells, and this could be eliminated by treatment with either DMSO or antioxidants. Our findings indicate that differential mechanisms involving NO and ROS signaling factors play a role in bystander responses generated from targeted T98G glioma and AG0 fibroblasts, respectively. These bystander interactions suggest that a mechanistic control of the bystander effect could be of benefit to radiotherapy. PMID- 15756684 TI - Cytotoxic effects of gammadelta T cells expanded ex vivo by a third generation bisphosphonate for cancer immunotherapy. AB - Nitrogen containing-bisphosphonates (N-BPs), widely used to treat bone diseases, have direct antitumor effects via the inactivation of Ras proteins. In addition to the direct antitumor activities, N-BPs expand gammadeltaT cells, which exhibit major histocompatibility complex-unrestricted lytic activity. BPs accumulate intermediate metabolites which may be tumor antigens in target cells. The purpose of our study was to clarify the cytotoxicity of gammadelta T cells expanded ex vivo by the most potent N-BP, zoledronate (ZOL). Especially, we focused on the importance of pretreatment against target cells also with ZOL; 1 microM ZOL plus IL-2 increased the absolute number of gammadeltaT cells 298-768 fold for 14 days incubation. The small cell lung cancer and fibrosarcoma cell lines pretreated with 5 microM ZOL showed a marked increase in sensitivity to lysis by gammadeltaT cells. While, untreated cell lines were much less sensitive to lysis by gdT cells. Video microscopy clearly demonstrated that gammadeltaT cells killed target cells pre-treated with ZOL within 3 hr. Pretreatment with 80 microg/kg ZOL also significantly enhanced the antitumor activity of gammadeltaT cells in mice xenografted with SBC-5 cells. These findings show that ZOL significantly stimulated the proliferation of gammadeltaT cells and that gammadeltaT cells required pre-treatment with ZOL for cytotoxic activity against target cells. PMID- 15756685 TI - Association of common ATM polymorphism with bilateral breast cancer. AB - The ATM kinase has an essential role in maintaining genomic integrity. Loss of both ATM alleles results in ataxia-telangiectasia (A-T), a rare autosomal recessive neuroimmunologic disorder associated with cancer susceptibility. Individuals heterozygous for germline ATM mutations have been reported to have an increased risk for malignancy, in particular, female breast cancer. In the current study, a full mutation analysis of the ATM gene was carried out in patients from 121 breast or breast-ovarian cancer families. We discovered that the combination of 5557G-->A in cis position with IVS38-8 T-->C was associated with bilateral breast cancer (OR = 10.2; 95% CI = 3.1-33.8; p = 0.001). As the 5557G-->A change has been reported to affect an exonic splicing enhancer, we hypothesized that the observed composite allele could have some effect on the correct splicing of exon 39. However, no aberrant transcripts were detected, but ATM expression levels of lymphoblast cell lines from heterozygous carriers of this combination allele were lower than from noncarriers (p = 0.09). Lowered gene expression levels may have direct influence on the activities in DNA damage recognition and response pathways, as well as other genome integrity maintenance functions. Based on the results, we propose a cancer risk-modifying effect for the ATM 5557G-->A, IVS38-8T-->C composite allele. PMID- 15756686 TI - Low level, long-term inorganic arsenite exposure causes generalized resistance to apoptosis in cultured human keratinocytes: potential role in skin co carcinogenesis. AB - Inorganic arsenic is a human carcinogen that targets the skin. Carcinogenesis is a multistep process in which acquired apoptotic resistance is a common event and prior work in non-skin cells shows acquired resistance to apoptosis occurs with chronic arsenite exposure. In the present study, when HaCaT cells, an immortalized, non-tumorigenic human keratinocyte cell line, were continuously exposed to low-level inorganic arsenite (as sodium arsenite; 100 nM) for 28 weeks, the cells acquired a generalized resistance to apoptosis. This included resistance to apoptosis induced by acute high concentrations of arsenite, ultraviolet A (UVA) irradiation, and several chemotherapeutic compounds (cisplatin, etoposide and doxorubicin). These arsenite-tolerant (As-TL) cells showed similar levels of UVA-induced reactive oxygen species (ROS) and oxidative DNA damage when compared to passage match control cells. Because cellular apoptosis is dependent on the balance between proapoptotic and survival pathways, the roles of protein kinase B (PKB), a key antiapoptotic molecule, in this acquired apoptotic resistance were investigated. Stimulation of apoptosis markedly decreased nuclear phosphorylated PKB (P-PKB) levels in control cells, but As-TL cells showed greatly increased stability of nuclear P-PKB. Pretreatment of the As-TL cells with LY294002 or Wortmannin, which specifically inhibit PKB phosphorylation, completely blocked apoptotic resistance in As-TL cells, indicating acquired apoptotic resistance is associated with increased stability of nuclear P-PKB. Because arsenic and UV irradiation are co-carcinogenic in mouse skin, resistance to UV-induced apoptosis in As-TL cells may allow UV-damaged cells to escape normal cell population controls and initiate the carcinogenic cascade. The observation that As-TL cells show no lessening of UV-induced genotoxicity supports this possibility. PMID- 15756687 TI - Hepatitis C infection is not associated with systemic HIV-associated non Hodgkin's lymphoma: a cohort study. AB - Immunosuppression induced by the human immunodeficiency virus (HIV) increases the risk of developing non-Hodgkin's lymphoma (NHL). As the hepatitis C virus (HCV) has been implicated in the development of B cell lymphomas, we compared the incidence of systemic NHL during HIV infection compared to HIV and HCV co infection. Of 5,832 individuals studied during the era of highly active anti retroviral therapy (HAART), 102 patients were diagnosed with systemic NHL. The incidence of systemic NHL was 6.9 of 10(4) patient years during HIV infection compared to 7.1 of 10(4) patient years during HIV alone (p = 0.9). In this immunocompromised patient population, there was no association between HCV infection and an increased risk of lymphoma. PMID- 15756689 TI - Coffee consumption and the risk of primary liver cancer: pooled analysis of two prospective studies in Japan. AB - Although case-control studies suggested that coffee consumption is associated with a decreased risk of liver cancer, no prospective cohort study has been carried out. To examine the association between coffee consumption and the risk of liver cancer, we conducted a pooled analysis of data available from 2 cohort studies in Japan. A self-administered questionnaire about the frequency of coffee consumption and other health habits was distributed to 22,404 subjects (10,588 men and 11,816 women) in Cohort 1 and 38,703 subjects (18,869 men and 19,834 women) in Cohort 2, aged 40 years or more, with no previous history of cancer. We identified 70 and 47 cases of liver cancer among the subjects in Cohort 1 (9 years of follow-up with 170,640 person-years) and Cohort 2 (7 years of follow-up with 284,948 person-years), respectively. We used Cox proportional hazards regression analysis to estimate the relative risk (RR) and 95% confidence interval (CI) of liver cancer incidence. After adjustment for potential confounders, the pooled RR (95% CI) of drinking coffee never, occasionally and 1 or more cups/day were 1.00 (Reference), 0.71 (0.46-1.09) and 0.58 (0.36-0.96), respectively (p for trend = 0.024). In the subgroup of subjects with a history of liver disease, we found a significant inverse association between coffee consumption and the risk of liver cancer. Our findings support the hypothesis that coffee consumption decreases the risk of liver cancer. Further studies to investigate the role of coffee in prevention of liver cancer among the high-risk population are needed. PMID- 15756690 TI - Increased plasma MMP9 in integrin alpha1-null mice enhances lung metastasis of colon carcinoma cells. AB - Inhibitors of matrix metalloproteinases (MMPs) were developed as anticancer agents based on the observation that MMPs facilitate local tumor spread and metastasis by promoting matrix degradation and cell migration. Unfortunately, these inhibitors were unsuccessful in the clinical treatment of several cancers, including lung cancer. A possible reason contributing to their failure is that MMP activity is critical for the generation of inhibitors of tumor angiogenesis, including angiostatin. Thus, MMPs might play opposing roles in tumor vascularization and invasion. To determine which effect of elevated MMP levels dominates in the progression of metastatic cancer, experimental lung metastasis assays were performed in integrin alpha1-null mice, a genetic model for increased plasma levels of MMP9 and MMP9-generated angiostatin (Pozzi et al., Proc. Natl. Acad. Sci. USA 2000;97:2202-7). We show that while the number of lung colonies in integrin alpha1-null mice was significantly increased compared to their wild-type counterparts, tumor volume was markedly reduced. In vivo treatment with the MMP inhibitor doxycycline resulted in a significant decrease in the number of lung colonies in both genotypes, but the tumors that formed were bigger and more vascularized. Increased tumor vascularization paralleled decreased plasma levels of MMP9 and consequent decreased angiostatin synthesis. These results demonstrate that while inhibition of MMPs prevents and/or reduces tumor invasion and lung metastasis, it has the paradoxical effect of increasing the size and vascularization of metastatic tumors due to decreased generation of inhibitors of endothelial cell proliferation. The continued growth of these large well vascularized tumors may explain the poor efficacy of MMP inhibitors in lung cancer clinical trials. PMID- 15756691 TI - Adenoviral vector-mediated gene transfer of IL-13Ralpha2 chain followed by IL-13 cytotoxin treatment offers potent targeted therapy for cytotoxin-resistant cancers. AB - Previous studies demonstrated that IL-13Ralpha2 chain-overexpressing cancer cells were highly sensitive to IL-13 cytotoxin (IL13-PE38QQR) and could be targeted by cytotoxin treatment. However, the majority of human tumors do not express high levels of IL-13Ralpha2 chain. To expand the IL-13 cytotoxin-mediated cancer targeting therapy, we combined cytotoxin treatment with gene transfer of IL 13Ralpha2 chain. We constructed a recombinant adenoviral vector carrying the human IL-13Ralpha2 gene (Ad-IL-13Ralpha2), which expresses high levels of IL 13Ralpha2 chain on infected cells. Human cancer cell lines A549 and HOS, which originally show no IL-13Ralpha2 expression and little sensitivity to IL-13 cytotoxin, were effectively converted to become sensitive to this cytotoxin after Ad-IL-13Ralpha2 infection. The CC(50) of IL-13 cytotoxin for Ad-IL-13Ralpha2 infected A549 cells was <10 ng/ml, whereas the CC(50) for uninfected or control vector-infected cells was >500 ng/ml. We also examined the antitumor activity of IL-13 cytotoxin in an established xenograft model of cytotoxin-resistant human lung tumor. Only a single i.t. injection of Ad-IL-13Ralpha2 markedly enhanced the sensitivity of established tumors to IL-13 cytotoxin treatment; furthermore, this antitumor effect was significantly sustained for more than 1 month after the last treatment with IL-13 cytotoxin. Taken together, these results suggest the combination of adenoviral vector-mediated IL-13Ralpha2 gene transfer and IL-13 cytotoxin administration can be an effective targeting approach for several types of IL-13 cytotoxin-resistant cancers which show no or little expression of IL 13Ralpha2 chain. PMID- 15756692 TI - A role for CXCR4 signaling in survival and migration of neural and oligodendrocyte precursors. AB - Oligodendrocyte development is controlled by a number of survival and migratory factors. The present study shows that signaling of CXCR4 receptor by the chemokine CXCL12 regulates survival and migration of neural precursors (NP) as well as oligodendrocyte progenitors (OP). CXCR4 is expressed by E14 striatal NP and OP generated by neurospheres. In CXCR4-defective mice, the number of NP in neurosphere outgrowth was twofold less than in wild-type (WT) mice; NP radial cell migration was also decreased. In contrast, the addition of CXCL12 to WT NP increased radial migration from the sphere in a dose-dependent manner with a maximal response at 200 nM. When oligodendrocytes differentiated in neurosphere outgrowth, CXCR4 was downregulated. OP isolated from newborn brain coexpressed CXCR4 with platelet-derived growth factor receptor-alpha (PDGFR alpha) or chondroitin sulfate proteoglycan; receptor expression also decreased during differentiation in vitro. Neonatal OP showed a peak migratory response to 20 nM of CXCL12 in chemotactic chambers, a migration inhibited by a CXCR4 antagonist and anti-CXCL12 antibody. In the embryonic spinal cord, the number of OP expressing PDGFR alpha was reduced more than twofold in CXCR4-defective mice compared with WT and the ratio of ventral to dorsal OP was significantly increased. This indicates a defect in OP survival and their dorsal migration from the ventral cord region, probably because CXCR4(-/-) OP are unable to respond to CXCL12 made by vascular endothelia and the pia mater. We propose that CXCR4 signaling regulate survival and outward chemotactic migration of OP during embryonic and postnatal CNS development. PMID- 15756693 TI - Ordered-subset analysis of savant skills in autism for 15q11-q13. AB - Autism is a complex disorder characterized by genetic and phenotypic heterogeneity. Analysis of phenotypically homogeneous subtypes has been used to both confirm and narrow potential autism linkage regions such as the chromosomal region 15q11-q13. Increased evidence for linkage in this region had been found in a subgroup of 21 autism families (total families = 94) stratified based on a savant skill factor (SSF) from the Autism Diagnostic Interview, Revised (ADI-R). We examined the savant phenotypic finding in our sample of 91 multiplex autism families. Using two-point parametric analysis in stratification with a cutoff point of a savant skill score of 0.16, our families failed to demonstrate linkage to 15q11-q13. In addition, ordered subset analysis (OSA) using SSF as a covariate also failed to show evidence for linkage. Our findings do not support savant skills as an informative phenotypic subset for linkage in our sample. PMID- 15756694 TI - Synthesis, in vitro-antimycobacterial activity and cytotoxicity of some alkyl alpha-(5-aryl-1, 3, 4-thiadiazole-2-ylthio)acetates. AB - A new series of alkyl alpha-[5-(5-nitro-2-furyl)-1, 3, 4- thiadiazole-2-ylthio] and alpha-[5-(1-methyl-5-nitro-2-imidazolyl)-1, 3, 4-thiadiazole-2 ylthio]acetates (6a-e, 6f-j) were synthesized and evaluated against Mycobacterium tuberculosis as part of the TAACF (Tuberculosis Antimicrobial Acquisition and Coordinating Facility) TB screening program. Primary screening was conducted at the single concentration of 6.25 microg/mL against M. tuberculosis H(37)Rv in BACTEC 12B medium using a broth microdilution assay, the Microplate Alamar Blue Assay (MABA). The minimum inhibitory concentration (MIC) was determined for compounds demonstrating >90 % growth inhibition in the primary screening. Seven compounds were efficient antimycobacterial agents showing MIC values ranging from 0.78 to 6.25 microg/mL. Among nitrofuran derivatives, methyl (6a), ethyl (6b), and benzyl (6e) esters displayed a good antituberculosis activity (MIC=0.78-3.13 microg/mL) and the others were inactive. In the nitro imidazole series, methyl (6f), ethyl (6g), propyl (6h) and butyl (6i) esters showed significant activity against M. tuberculosis while benzyl (6j) ester was inactive. Also, active compounds were screened by serial dilution to assess toxicity to a VERO cell line. A varying degree of toxicity was observed in nitrofuran and nitroimidazole derivatives (IC(50) = 2.3 - >10 microg/mL). PMID- 15756695 TI - Synthesis, analgesic and antiinflammatory properties of certain 5-/6-acyl-3-(4 substituted-1-piperazinylmethyl)-2-benzoxazolinones derivatives. AB - The synthesis of a novel series of Mannich bases of 5-/6-acyl-5-methyl-2 benzoxazolinones has been described. The structures attributed to compounds 2a, 3a, 4a, 4b, 9a, 9b, 5a-5g, 6a-6g, 10a, 10g, 11a, 11g have been elucidated using IR and (1)H NMR spectroscopic techniques besides elemental analysis. The compounds have been evaluated for their in vivo analgesic and antiinflammatory activities using the p-benzoquinone-induced writhing test and the carrageenan hind paw oedema test in mice, respectively. In addition, the ulcerogenic effects of the compounds were determined. Among the tested derivatives most promising results were obtained for the compounds bearing a 6-(4-chlorobenzoyl) at C-6 position and 2-/4-fluorophenyl at C-3 position of 2-benzoxazolinone ring (11c, 11d). PMID- 15756696 TI - Smart polyion complex micelles for targeted intracellular delivery of PEGylated antisense oligonucleotides containing acid-labile linkages. AB - A novel pH-sensitive and targetable antisense ODN delivery system based on multimolecular assembly into polyion complex (PIC) micelles of poly(L-lysine) (PLL) and a lactosylated poly(ethylene glycol)-antisense ODN conjugate (Lac-PEG ODN) containing an acid-labile linkage (beta-propionate) between the PEG and ODN segments has been developed. The PIC micelles thus prepared had clustered lactose moieties on their peripheries and achieved a significant antisense effect against luciferase gene expression in HuH-7 cells (hepatoma cells), far more efficiently than that produced by the nonmicelle systems (ODN and Lac-PEG-ODN) alone, as well as by the lactose-free PIC micelle. In line with this pronounced antisense effect, the lactosylated PIC micelles showed better uptake than the lactose-free PIC micelles into HuH-7 cells; this suggested the involvement of an asialoglycoprotein (ASGP) receptor-mediated endocytosis process. Furthermore, a significant decrease in the antisense effect (27 % inhibition) was observed for a lactosylated PIC micelle without an acid-labile linkage (thiomaleimide linkage); this suggested the release of the active (free) antisense ODN molecules into the cellular interior in response to the pH decrease in the endosomal compartment is a key process in the antisense effect. Use of branched poly(ethylenimine) (B-PEI) instead of the PLL for PIC micellization led to a substantial decrease in the antisense effect, probably due to the buffer effect of the B-PEI in the endosome compartment, preventing the cleavage of the acid-labile linkage in the conjugate. The approach reported here is expected to be useful for the construction of smart intracellular delivery systems for antisense ODNs with therapeutic value. PMID- 15756697 TI - "Turn ON/OFF your LOV light": boron-dipyrromethene-flavin dyads as biomimetic switches derived from the LOV domain. PMID- 15756698 TI - Resistance of insect pests to neonicotinoid insecticides: current status and future prospects. AB - The first neonicotinoid insecticide introduced to the market was imidacloprid in 1991 followed by several others belonging to the same chemical class and with the same mode of action. The development of neonicotinoid insecticides has provided growers with invaluable new tools for managing some of the world's most destructive crop pests, primarily those of the order Hemiptera (aphids, whiteflies, and planthoppers) and Coleoptera (beetles), including species with a long history of resistance to earlier-used products. To date, neonicotinoids have proved relatively resilient to the development of resistance, especially when considering aphids such as Myzus persicae and Phorodon humuli. Although the susceptibility of M. persicae may vary up to 20-fold between populations, this does not appear to compromise the field performance of neonicotinoids. Stronger resistance has been confirmed in some populations of the whitefly, Bemisia tabaci, and the Colorado potato beetle, Leptinotarsa decemlineata. Resistance in B- and Q-type B. tabaci appears to be linked to enhanced oxidative detoxification of neonicotinoids due to overexpression of monooxygenases. No evidence for target site resistance has been found in whiteflies, whereas the possibility of target site resistance in L. decemlineata is being investigated further. Strategies to combat neonicotinoid resistance must take account of the cross-resistance characteristics of these mechanisms, the ecology of target pests on different host plants, and the implications of increasing diversification of the neonicotinoid market due to a continuing introduction of new molecules. PMID- 15756699 TI - Pyridalyl, a novel insecticide: potency and insecticidal selectivity. AB - Pyridalyl is an insecticide of a novel chemical class (unclassified insecticides). Toxicity of pyridalyl to two insect pest species, Spodoptera litura and Frankliniella occidentalis, an insect predator, Orius stringicollis, and a pollinator, Bombus terrestris, was evaluated in the laboratory. The insecticidal activity of pyridalyl against S. litura was evaluated using the leaf dipping method. The potency of pyridalyl was highly effective against all development stages (2nd to 6th instar larvae) of S. litura. This compound was also evaluated against F. occidentalis using the direct spray method. For F. occidentalis, toxicity of pyridalyl was almost similar to that of acrinathrin, but greater than acrinathrin for adults. Then the toxicity of this product to the natural enemies, Orius stringicollis and the pollinating insect Bombus terrestris, was evaluated using the body-dipping method or direct spray method. No acute toxicity of this product was observed on these non-target insects. Moreover, the influence of pyridalyl to the nest of Bombus terrestris was evaluated using the direct spray to the inside of the nest. No apparent influence of this compound was observed by 21 days after treatment. The cytotoxicity of pyridalyl to the Sf9 insect cell line and the CHO-K1 mammalian cell line was evaluated using the trypan-blue exclusion method. High toxicity to the insect cell line, but almost no toxicity to the mammalian cell line, was observed. Thus, pyridalyl exhibited high selectivity in cytotoxicity between the insect and mammalian cell line as well as in insecticidal activity among insect species. We infer pyridalyl may be useful for IPM programs of greenhouse cultivation system. PMID- 15756700 TI - Biochemical mode of action and differential activity of new ecdysone agonists against mosquitoes and moths. AB - THQ (1-aroyl-4-(arylamino)-1,2,3,4-tetrahydroquinoline) compounds were identified by FMC Corporation in cell-based assays that used ecdysone receptors from Drosophila melanogaster, Heliothis virescens, or Plodia interpunctata. THQ compounds showed weak insecticidal activity against H. virescens and, therefore, were not developed further. Several ecdysone agonists based on THQ chemotype have been synthesized and tested for their activity against a number of EcRs in transactivation assays. The THQ compound, RG-120768, activated AaEcR (EcR from A. aegypti) but did not activate EcRs cloned from other insects. In transactivation assays, all six THQ ligands tested functioned through AaEcR but not through CfEcR (EcR from Choristoneura fumiferana). Three THQ compounds that showed higher activity in transactivation assays were tested in tobacco bud moth, H. virescens, and yellow fever mosquito, A. aegypti. These compounds showed higher activity in A. aegypti when compared to their activity in H. virescens. These data show that the THQ ligands are a new class of non-steroidal ecdysone agonists with preferential activity against mosquitoes. PMID- 15756702 TI - Biorational insecticides: mechanism and cross-resistance. AB - Potency and cross-resistance of various biorational insecticides, exemplified by the whitefly Bemisia tabaci, have been studied. Bemisia tabaci were exposed to the juvenile hormone mimic pyriproxyfen for the past 12 years resulting in an over 2,000-fold resistance, but there was no appreciable cross-resistance with the benzoylphenyl urea novaluron. Similarly, no cross-resistance was found between pyriproxyfen and the two neonicotinoids, acetamiprid and imidacloprid. On the other hand, a slight cross-resistance of 5-13-fold was observed with another neonicotinoid thiamethoxam. Among the neonicotinoids, a resistant strain of B. tabaci to thiamethoxam (approximately 100-fold) showed no appreciable cross resistance to either acetamiprid or imidacloprid, while another strain 500-fold resistant to thiamethoxam resulted in a mild of 4-6-fold resistance to acetamiprid and imidacloprid. In other assays, B. tabaci strain resistant to thiamethoxam (approximately 100-fold) had no cross-resistance to pyriproxyfen. Our findings indicate that no appreciable cross-resistance was observed between the benzoylphenyl urea novaluron, the juvenile hormone mimic pyriproxyfen, and the neonicotinoids acetamiprid and imidacloprid. Hence, these compounds could be used as components in insecticide resistance management programs. PMID- 15756703 TI - Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance. AB - Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied. PMID- 15756704 TI - Relationship between renal anemia and prognostic stages of IgA nephropathy. AB - In 2002, the Joint Committee of the Special Study Group on Progressive Glomerular Diseases, Ministry of Health, Labor and Welfare of Japan newly revised the clinical guidelines for IgA nephropathy (Sakai et al.: Jpn J Nephrol 37:417-421, 1995; Tomino and Sakai: Clin Exp Nephrol, 7, 93-97, 2003). The prognostic stages were classified into four groups: the good prognosis group (Group I), relatively good prognosis group (Group II), relatively poor prognosis group (Group III), and poor prognosis group (Group IV). The relationship between the levels of Hb, Ht, and RBC in peripheral blood and the renal prognostic stages was determined in 62 patients with IgA nephropathy in the present study. The mean levels of Hb, Ht, and RBC were significantly lower in Group IV than in Group I (P<0.05). However, there were no significant changes in the levels of serum creatinine (s-Cr) or creatinine clearance (CCr) among these four groups. It appears that the levels of Hb, Ht, and RBC in peripheral blood may be important clinical parameters for the evaluation of prognostic stages in patients with IgA nephropathy. PMID- 15756705 TI - AT1 receptor A1166C and AT2 receptor -1332A/G gene polymorphisms: efficient genotyping by single-tube PCR. AB - Angiotensin II type 1 receptor (AT1) and angiotensin II type 2 receptor (AT2) genes have been investigated in recent years as potential etiologic candidates for cardiovascular and renal diseases. The pathogenic implications of AT1 A1166C and AT2 A-1332G gene polymorphisms have been shown. Here we describe a rapid and reliable method for detecting both AT1 and AT2 gene polymorphisms by a single tube PCR, to reduce analysis time and simplify the genotyping procedure. In contrast to previously described methods, our method does not require hybridization, primer extension, or nested PCR for genotyping. In most previous studies concerning gene polymorphisms of RAS, both AT1 and AT2 receptor gene polymorphisms were investigated. The advantage of our method is that it makes it possible to detect both of these polymorphisms in a duplex PCR. The procedure described is convenient for routine laboratory use with manual sample processing, and offers the potential for further automation as well. Its simplicity makes it practical for large-scale screening of individuals and families at risk for cardiovascular or renal diseases. PMID- 15756706 TI - Analytical performance evaluation of a new turbidimetric immunoassay for valproic acid on the ADVIA 1650 analyzer: effect of gross hemolysis and high bilirubin. AB - Valproic acid is an anticonvulsant that requires careful therapeutic drug monitoring. Valproic acid is also used in psychiatric patients. Bayer Diagnostics (Tarrytown, NY) recently marketed a turbidimetric immunoassay for monitoring valproic acid concentrations in serum or plasma using the ADVIA 1650 analyzer. We evaluated the performance of this new assay by comparing it with a widely used fluorescence polarization immunoassay (FPIA) on the AxSYM analyzer (Abbott Laboratories, Abbott Park, IL). The total coefficient of variation (CV) for the low control of this new assay was 6.8% (mean = 30.7, SD = 2.1 microg/mL, n = 44) while the corresponding CVs for the medium and high controls were 3.3% (mean = 81.0, SD = 2.7 microg/mL, n = 44) and 5.9% (mean = 142.9, SD = 8.4 microg/mL, n = 44), respectively. The assay is linear up to a serum valproic acid concentration of 170 microg/mL, and the detection limit is 4.4 microg/mL. We observed an excellent correlation between the FPIA of valproic acid and the turbidimetric assay using specimens from 52 different patients who were receiving valproic acid. Using the valproic acid concentrations obtained by the FPIA as the x-axis, and the corresponding valproic acid concentrations obtained by the turbidimetric assay as the y-axis, we developed the following regression equation: y = 1.03 x+1.55 (r = 0.98). With this new assay, high concentrations of bilirubin (unconjugated 30 mg/dL and conjugated 30 mg/dL) and gross hemolysis (4+, hemoglobin: 1,500 mg/dL) have no effect on measurements of valproic acid concentration. We conclude that the new turbidimetric assay for valproic acid can be used for routine therapeutic drug monitoring of valproic acid in clinical laboratories. PMID- 15756707 TI - Pleural fluid interferon-gamma and adenosine deaminase levels in tuberculosis pleural effusion: a cost-effectiveness analysis. AB - Pleural fluid levels of interferon-gamma (IFN-gamma) and adenosine deaminase (ADA) have been found to be high in patients with tuberculosis (TB). The present study was carried out to compare the diagnostic utility of these two markers and to carry out a cost-effectiveness analysis of performing IFN-gamma estimation in comparison to ADA. A total of 52 patients with pleural effusion, 35 of which were found to have TB etiology, were prospectively included for estimation of ADA and IFN-gamma levels. The difference in the cost of performing the two diagnostic tests was compared with the cost of the treatment for a patient with TB. Pleural fluid IFN-gamma (median [range]: 2,100 [70-14,000] vs. 3 [0-160]; P<0.001) as well as ADA levels (mean [SD]: 93.1 [62.3] vs 15.4 [8.7]; P<0.001) were significantly higher in patients with TB effusion. Even though IFN-gamma estimation was more sensitive (97.1 vs. 91.4%), the extra cost of IFN-gamma estimation for detecting one patient with TB was found to be equivalent to the cost of a complete course of antituberculosis treatment for six patients. In developing countries, where TB is rampant and cost is a major concern, pleural fluid IFN-gamma estimation does not seem to be a cost-effective investigation method for differentiating TB from non-TB pleural effusion. PMID- 15756708 TI - Hematologic aspects of myeloablative therapy and bone marrow transplantation. AB - The transplantation of bone marrow cells or isolated hematopoietic stem cells from the bone marrow or peripheral blood is a widely utilized form of therapy for patients with incurable diseases of the hematopoietic and immune systems. Successful engraftment of the transplanted stem cells in an adequately prepared recipient normally leads to bone marrow reconstitution over a period of several weeks, accompanied by more gradual reconstitution of the immune system. Since the recipient is profoundly ill during the initial treatment period, laboratory data is critical for monitoring engraftment, detecting residual/recurrent disease, and identifying problems that may delay bone marrow reconstitution or lead to other medical complications. Accurate blood cell counts are imperative, and most bone marrow transplantation patients undergo periodic monitoring with bone marrow aspirates and biopsies with cytogenetic, molecular, and multiparametric flow cytometric studies. The potential complications of bone marrow transplantation include engraftment failure and delayed engraftment, infection, residual bone marrow disease, acute and chronic graft versus host disease, myelofibrosis, therapy-related acute leukemia, post-transplant lympho-proliferative disorders, and toxic myelopathy. PMID- 15756709 TI - Distribution of oxidation enzyme eNOS and myeloperoxidase in primary open angle glaucoma. AB - Genetic factors and the influence of superoxide are known to play roles in the etiology of glaucoma. We evaluated the association between primary open angle glaucoma (POAG) and two polymorphisms in the epithelial nitric oxide synthase (eNOS) gene, and one polymorphism in the myeloperoxidase (MPO) gene. We enrolled 66 patients with POAG and 100 healthy volunteers in this study. The polymorphisms in the eNOS and the polymorphism MPO -463 G-to-A in the MPO gene were resolved by polymorphism polymerase chain reaction (PCR). There were no significant differences in the distribution of the eNOS intron -4 (P=0.481), eNOS promotor 786 (P=0.555), and MPO -463 (P=0.292) gene polymorphisms between the POAG patients and the volunteers (P-values=0.481, 0.555, and 0.292, respectively). None of the alleles from either gene differed between the groups (P-values=0.483, 0.554, and 0.183, respectively). Superoxide is closely related to glaucoma, and eNOS and MPO are two important enzymes in the free radical pathway. However, polymorphisms of the eNOS intron-4, eNOS promotor -786, and MPO -463 gene polymorphisms did not reveal significant differences between POAG patients and controls in our study. The use of these agents and other superoxide-related genes for clinical applications requires further investigation. PMID- 15756710 TI - Serum levels of neopterin and interleukin-2 receptor in women with severe preeclampsia. AB - Preeclampsia continues to be a major cause of maternal and perinatal mortality and morbidity worldwide. The etiopathogenesis of preeclampsia is not fully understood. Neopterin and interleukin-2 (IL-2) production reflects cellular immunity. Our purpose was to determine the levels of neopterin and interleukin-2 receptor (IL-2R) in pregnant women with severe preeclampsia, and assess the implications of these findings in the pathophysiology of preeclampsia. Fourteen women with preeclampsia were compared with 14 healthy pregnant women. Serum levels of neopterin were measured by an enzyme-linked immunoassay (ELISA), and IL 2R levels were determined by an immunoassay method with an Immulite analyzer. The levels of neopterin and IL-2R were significantly higher in the preeclamptic subjects than in normotensive women (P<0.05). There was a significant correlation between neopterin and IL-2R. We found that serum neopterin and IL-2R levels are increased in women with severe preeclampsia. The results of this study suggest that a T-helper 1 (Th1) type immune mechanism is involved in the pathogenesis of preeclampsia. PMID- 15756711 TI - Replication-dependent transgene expression from a conditionally replicating adenovirus via alternative splicing to a heterologous splice-acceptor site. AB - BACKGROUND: Oncolytic viruses are promising anticancer agents because they selectively kill cancer cells and multiply within a tumor. Their oncolytic potency might be improved by expressing a therapeutic gene from the virus genome. In this regard, proper kinetics and level of transgene expression are important. In addition, expression of cytotoxic transgene products should be confined to cancer cells. Here, we developed oncolytic adenoviruses that provide transgene expression dependent on viral replication. METHODS: We constructed an oncolytic adenovirus that expresses luciferase under regulation of the endogenous major late promoter (MLP) via alternative splicing to an inserted splice-acceptor site analogous to that of the adenovirus serotype 40 long fiber gene. Splicing of the luciferase transcript was studied by RT-PCR analysis. Expression was measured in the presence and absence of the flavonoid apigenin, an inhibitor of viral replication. RESULTS: The inserted splice-acceptor site was properly recognized by the adenoviral splicing machinery. Luciferase expression levels were markedly higher than levels obtained with the cytomegalovirus (CMV) promoter, especially at late stages of infection. Inhibiting adenovirus replication reduced luciferase expression levels dramatically by 4 to 5 logs, whereas expression levels with the CMV-luciferase adenovirus were only moderately affected (2 logs). CONCLUSIONS: Transgene delivery using the endogenous late gene expression machinery resulted in an expression pattern distinct from expression driven by the conventional CMV promoter. The high expression levels and strict coupling of expression to viral replication should be useful for adequate monitoring of replication and might provide a platform for the design of armed conditionally replicating adenoviruses (CRAds) with enhanced oncolytic potency. PMID- 15756712 TI - The use of absorption enhancers to enhance the dispersibility of spray-dried powders for pulmonary gene therapy. AB - BACKGROUND: Pulmonary gene therapy requires aerosolisation of the gene vectors to the target region of the lower respiratory tract. Pulmonary absorption enhancers have been shown to improve the penetration of pharmaceutically active ingredients in the airway. In this study, we investigate whether certain absorption enhancers may also enhance the aerosolisation properties of spray-dried powders containing non-viral gene vectors. METHODS: Spray-drying was used to prepare potentially respirable trehalose-based dry powders containing lipid-polycation-pDNA (LPD) vectors and absorption enhancers. Powder morphology and particle size were characterised using scanning electron microscopy and laser diffraction, respectively, with gel electrophoresis used to assess the structural integrity of the pDNA. The biological functionality of the powders was quantified using in vitro cell (A549) transfection. Aerosolisation from a Spinhaler dry powder inhaler into a multistage liquid impinger (MSLI) was used to assess the in vitro dispersibility and deposition of the powders. RESULTS: Spray-dried powder containing dimethyl-beta-cyclodextrin (DMC) demonstrated substantially altered particle morphology and an optimal particle size distribution for pulmonary delivery. The inclusion of DMC did not adversely affect the structural integrity of the LPD complex and the powder displayed significantly greater transfection efficiency as compared to unmodified powder. All absorption enhancers proffered enhanced powder deposition characteristics, with the DMC-modified powder facilitating high deposition in the lower stages of the MSLI. CONCLUSIONS: Incorporation of absorption enhancers into non-viral gene therapy formulations prior to spray-drying can significantly enhance the aerosolisation properties of the resultant powder and increase biological functionality at the site of deposition in an in vitro model. PMID- 15756713 TI - Rad51 siRNA delivered by HVJ envelope vector enhances the anti-cancer effect of cisplatin. AB - BACKGROUND: Every cancer therapy appears to be transiently effective for cancer regression, but cancers gradually transform to be resistant to the therapy. Cancers also develop machineries to resist chemotherapy. Short interfering RNA (siRNA) has been evaluated as an attractive and effective tool for suppressing a target protein by specifically digesting its mRNA. Suppression of the machineries using siRNA may enhance the sensitivity to chemotherapy in cancers when combined with an effective delivery system. METHODS: To enhance the anti-cancer effect of chemotherapy, we transferred siRNA against Rad51 into various human cancer cells using the HVJ (hemagglutinating virus of Japan, Sendai virus) envelope vector in the presence or absence of cis-diamminedichloroplatinum(II) (CDDP, cisplatin). The inhibition of cell growth was assessed by a modified MTT assay, counting cell number, or fluorescence-activated cell sorting (FACS) analysis after Annexin V labeling. The synthetic Rad51 siRNA was also introduced into subcutaneous tumor masses of HeLa cells in SCID mice with or without intraperitoneal injection of CDDP, and tumor growth was monitored. RESULTS: When synthetic Rad51 siRNA was delivered into HeLa cells using the HVJ envelope vector, no Rad51 transcripts were detected on day 2, and Rad51 protein completely disappeared for 4 days after siRNA transfer. When HeLa cells were incubated with 0.02 microg/ml CDDP for 3 h after siRNA transfer, the number of colonies decreased to approximately 10% of that with scrambled siRNA. The sensitivity to CDDP was enhanced in various human cancer cells, but not in normal human fibroblasts. When Rad51 siRNA was delivered into tumors using the HVJ envelope vector, the Rad51 transcript level was reduced to approximately 25%. Rad51 siRNA combined with CDDP significantly inhibited tumor growth when compared to siRNA or CDDP alone. CONCLUSIONS: Rad51 siRNA could enhance the sensitivity to CDDP in cancer cells both in vitro and in vivo. Our results suggest that the combination of CDDP and Rad51 siRNA will be an effective anti-cancer protocol. PMID- 15756714 TI - A QSAR-modeling perspective on cationic transfection lipids. 1. Predicting efficiency and understanding mechanisms. AB - BACKGROUND: As gene therapy using viral vectors involves clinical risks, limited DNA-carrying capacity, and manufacturing problems, non-viral vectors, including cationic lipids, have been investigated. Unfortunately, these agents have significantly lower transfectional ability and, due to the complexity of the transfectional pathway, no general schemes exist for correlating cationic lipid chemistry with transfectional efficacy. METHODS: Quantitative structure-activity relationship (QSAR) analyses were carried out on sets of routinely used, experimental, and unsuccessful cationic lipid vectors taken from the literature. This approach described the amphipathic character, basicity, headgroup size, lipophilicity and shape of cationic lipids using numerical parameters. Compounds were plotted onto various parameter diagrams, and correlations were sought between numerical parameters and transfectional efficiency. RESULTS: Transfectionally effective cationic lipids fell into restricted zones in various parameter spaces, indicating that amphipathic character, lipid shape and lipophilicity were generally significant factors, whilst basicity and headgroup size were only important for certain compounds. The data supported the general significance of membrane mixing followed by induction of membrane curvature, and the more limited role of osmotic shock, as mechanisms of membrane disruption. QSAR descriptions of effective lipids permitted detailed chemical guidelines for optimizing cationic lipid structure to be given. Limitations of the approach and models are discussed. CONCLUSIONS: QSAR modeling indicated that induction of membrane curvature and osmotic shock are important mechanisms for membrane disruption by cationic lipids. The models also allowed specification of chemically detailed guidelines for selection or design of optimal cationic lipids. PMID- 15756716 TI - Effective repetitive dystrophin gene transfer into skeletal muscle of adult mdx mice using a helper-dependent adenovirus vector expressing the coxsackievirus and adenovirus receptor (CAR) and dystrophin. AB - BACKGROUND: The helper-dependent adenovirus (HDAd) vector is less immunogenic and has a larger cloning capacity of up to 37 kb enough to carry the full-length dystrophin cDNA. However, high and long-term expression of dystrophin transduced to mature muscle still remains difficult. One of the main reasons for this is that the expression of the coxsackievirus and adenovirus receptor (CAR) is very low in mature muscle. METHODS: We have constructed two different HDAd vectors. One contains the LacZ and the murine full-length dystrophin expression cassette (HDAdLacZ-dys), and the other is a new, improved vector containing the CAR and the dystrophin expression cassette (HDAdCAR-dys). RESULTS: We initially demonstrated high dystrophin expression and prevention of the dystrophic pathology in mdx muscle injected during the neonatal phase with HDAdLacZ-dys. Furthermore, we demonstrated that repeated injections of HDAdCAR-dys into mature muscle led to approximately nine times greater dystrophin-positive fibers in number than a single injection, thereby recovering the expression of dystrophin associated proteins. This data has also shown that HDAdCAR-dys enabled administration of adenovirus (Ad) vector to the host with pre-existing immunity to the same serotype of Ad. CONCLUSIONS: Repetitive injections of the HDAd vector containing the CAR and the dystrophin expression cassette could improve the efficiency of subsequent dystrophin gene transfer to mature mdx muscle. This result suggests that our new HDAd vector will provide a novel gene therapy strategy for Duchenne muscular dystrophy, raising the prospects for gene therapy of other hereditary myopathies. PMID- 15756717 TI - Nursing. HIV pain management: understanding the essentials of chronic pain assessment. PMID- 15756718 TI - The merits of blastocyst versus cleavage stage embryo transfer: a Cochrane review. PMID- 15756719 TI - Financial management--part II. PMID- 15756722 TI - Mental health care for India's tsunami survivors. PMID- 15756720 TI - Cystic lymphangioma of the upper extremity: US and MRI correlation (2004:11b). AB - The authors described a very rare case of the cystic lymphangioma (CL) of the left upper extremity in a 12-year-old patient. The diagnosis was made by US and MRI and confirmed by histological examination of the resected specimen. PMID- 15756723 TI - Cleaning up after the tsunamis. PMID- 15756724 TI - A crucial time for Afghanistan's fledgling health system. PMID- 15756725 TI - Avian influenza: perfect storm now gathering? PMID- 15756726 TI - [Neuromuscular diseases (NME). I. Spinal muscular atrophy, peripheral nerve diseases, congenital myasthenia gravis]. AB - Among the group of clinically and genetically heterogeneous spinal muscular atrophies(SMA), the autosomal recessive proximal types I-III are the most frequent. They are caused by mutations of the telomeric copy of the survival motor neuron gene (SMN1) on chromosome sq while loss of the centromeric copy (SMN2) does not lead to SMA. The conservation of exon 7 in the SMN2 copy seems to be crucial for possible causal therapy options. The genetic defect is also known for spinal muscular atrophy with respiratory distress. Hereditary polyneuropathies are caused by mutations in several genes on different chromosomes. Genetic and phenotypic heterogeneity must be considered. Data on clinical symptoms, inheritance, and neurophysiology are obligatory for the most effective molecular analysis. Hereditary congenital myasthenic syndromes are clinically, genetically, and pathogenetically heterogeneous. The diagnostic significance of molecular genetic analyses is still increasing. Therapeutic options include oral medication and assisted ventilation as needed. PMID- 15756727 TI - Peyton Rous: father of the tumor virus. AB - In 1910, Peyton Rous identified a transmissible avian tumor virus, a discovery that began the journey from tumor virus biology to tumor biology itself. PMID- 15756728 TI - [Increased carcinoembryonic antigen level: which diagnosis?]. PMID- 15756729 TI - [Kawasaki syndrome: a coronavirus infection?]. PMID- 15756730 TI - Tool for pain assessment. PMID- 15756732 TI - Teaching clinical ethics using a case study family presence during cardiopulmonary resuscitation. PMID- 15756731 TI - Risk evaluation in action for cardiovascular health. PMID- 15756733 TI - Capgras syndrome in the modern era: self misidentification on an ID picture. PMID- 15756734 TI - Family-oriented rehabilitation for unexplained chronic pain. PMID- 15756735 TI - Quetiapine therapy for corticosteroid-induced mania. PMID- 15756736 TI - Reader shares Web resources about unusual cravings. PMID- 15756737 TI - [Thanks to the team from Umea!]. PMID- 15756738 TI - [The need for persistent help]. PMID- 15756739 TI - [American military physicians participate in torture-like deeds]. PMID- 15756740 TI - EMR vs. DIMS. Nay: DIMS effective, less costly than EMR and easier to use. PMID- 15756741 TI - Stent competition helps prices. PMID- 15756742 TI - NY: Psychiatric patient 'refuses' to take lithium: nurse practitioner testifies re need for court order. PMID- 15756744 TI - In that case: two-year-old Sarah is admitted to the paediatric ward with acute myeloblastic leukaemia. Commentary. PMID- 15756745 TI - In that case: two-year-old Sarah is admitted to the paediatric ward with acute Myeloblastic leukaemia. Commentary. PMID- 15756747 TI - WHI memory study fails to find oestrogen benefit in dementia. PMID- 15756746 TI - Relationship between abdominal visceral fat and lacunar infarcts in Japanese men. PMID- 15756749 TI - [History of Czech physiology]. PMID- 15756748 TI - Physics applications in nuclear medicine: progress on many fronts. PMID- 15756750 TI - [Management of severe phenol burn combined with poisoning: report of one case]. PMID- 15756751 TI - A look on the bright side. PMID- 15756752 TI - 2004 approvals: the demise of the blockbuster? PMID- 15756753 TI - Iressa failure raises fears about accelerated approvals. PMID- 15756754 TI - Patent infringement. PMID- 15756755 TI - An audience with...Sir John Sulston. PMID- 15756756 TI - Female sexual dysfunction. PMID- 15756757 TI - Natalizumab. PMID- 15756758 TI - Cortical function: a view from the thalamus. AB - Neuroscientists from across the country gathered at the University of Wisconsin, Madison in September to honor Ray Guillery and his seminal work on the thalamus. The meeting focused on three timely research topics, each of which inspired new thinking about thalamic function. Presentations on the organization and dynamic nature of thalamocortical pathways, the role of the thalamus in communication between cortical areas, and the relationship between sensory and motor pathways of the brain, including cognitive aspects of thalamocortical processing, made for lively discussions. The meeting revealed that communication between thalamus and cortex is so rich that we should no longer consider the operations of either structure separately from the other. Proceedings of the meeting will be published in Progress in Brain Research in 2005. In this report, we provide a general overview of the main themes of the meeting. PMID- 15756763 TI - Death query. PMID- 15756764 TI - More cleaners are needed. PMID- 15756765 TI - New departments address disability issues. PMID- 15756766 TI - Dyslexia. PMID- 15756767 TI - Stem cell research. PMID- 15756768 TI - [Clinical application of Astragali radix]. PMID- 15756769 TI - Health savings accounts. PMID- 15756770 TI - A center of a different stripe. PMID- 15756771 TI - Lung cancer in females: an epidemic foretold. PMID- 15756772 TI - Interview on functional appliances. PMID- 15756773 TI - Screening for ovarian cancer: recommendation statement. U.S. Preventive Services Task Force. PMID- 15756774 TI - Physicians face sharp Medicare payment cuts beginning January 1, 2006. PMID- 15756775 TI - Dialogue. PMID- 15756776 TI - Monitoring for metabolic complications associated with atypical antipsychotics. PMID- 15756777 TI - State of South Dakota's Child 2004 continued: out of home care for infants. AB - South Dakota leads the nation in its percent of women in the workforce who have children under the age of six. Nationally, 64% of women with children this young are employed and this is the case for 78% of women in the state. Further, 60% of South Dakota's infants live in homes where either their single mother is employed or both their parents are employed outside of their home. Nearly half of all the state's infants and children under six receive nonparental care during their parents' working hours. The significance of this reality is profound as young children's experiences, during early formative periods of life, affect their current and future development. The dimensions of quality care for infants are described as well as the economic and policy dynamics that affect its delivery in South Dakota. PMID- 15756778 TI - Update on colorectal cancer screening in South Dakota. PMID- 15756779 TI - [Dilemmas and controversies related to cancers of the anterior laryngeal commissure]. AB - From diagnostic and therapeutical aspect, the cancers of the anterior laryngeal commissure are the separate category in glottic cancers. But, they have not been individually classified in the majority of statistical reports, and, therefore, the incidence of the anterior commissural cancer should be taken with precaution. The issue of therapeutical strategy is controversial, considering the options and limitations of resections of the anterior commissural tumors, within the conception of oncological radicalism. Dilemmas are being especially faced with in radiotherapy, given the failures and unsatisfactory radiotherapeutical results. The prospective clinical study included the analysis of the incidence of primary and secondary cancers of the anterior commissure of the larynx. The follow-up of five-year survivals allowed for the establishment of efficiency of the applied therapeutical methods. The results of five-year survival in patients treated by primary surgery were highly more significant in relation to results obtained by radiotherapy of patients. PMID- 15756780 TI - [Onco-surgical significance of anatomo-morphological specificity of glottic region of the larynx]. AB - Glottis is a medium level of the larynx, involving the vocal cords, vocal process of arytenoid cartilage, and the anterior and posterior commissures. This region of heterogeneous histomorphological structure has specific characteristics: the internal striated muscles, the outer and inner perichondrium, atypical intra cartilaginous areas of ossification, and unusual muscular insertion to cartilage. Microtomy of 3 thick successive sections was performed at the level of the upper surface of vocal cords. Standard histological staining methods were used. Microphotographs were taken by light microscope under different magnification along with histomorphometric measurements. Cancer spread is partially restricted by anatomic barriers: vocal ligament and tendon of the anterior commissure, and subsequently by the elastic cone. Easier tumor extension may be due to absence of the internal perichondrium and ossification of thyroid cartilage. Microvascularization of the anterior commissure is significant for tumor spread in glottis and paraglottic space. The role of commissural lymph network in local spread of the cancer is completely vague. It is certain that there are causes, still unrecognized, which have an effect on the pathways and direction of malignant tumor spread. PMID- 15756781 TI - [Extended supra glottic laryngectomies]. AB - In the period 1976-1998, 408 patients with supraglottic laryngeal cancer were primarily treated by surgery using the method of supraglottic laryngectomy. Classical supraglottic laryngectomy was performed in 355 patients, while 53 underwent extended supraglottic laryngectomy. The objective of the study was to analyze the success of the extended supraglottic laryngectomy in relation to classical supraglottic laryngectomy in indicated cases. T1 tumor was found in 171 (42%) patients, T2 in 212 (52%), while T3 was found in 25 (6%) cases. Local recurrence developed in 3 out of 53 patients operated by the extended supraglottic laryngectomy technique, and in 17 out of 355 operated by classical supraglottic laryngectomy (chi2=0.075, DF=1, p=0.784; Yates=0.00, p=1.00). Five year survival of patients operated by the extended supraglottic laryngectomy was reported in 40 out of 53 patients, while survival of patients operated by classical supraglottic laryngectomy was noted in 270 out of 355 cases (chi2=0.004, DF=1, p=0.926; Yates=0.00, p=1.00). There was no significant difference of local recurrence and five-year survival between patients treated by classical and extended supraglottic laryngectomy. PMID- 15756782 TI - [Surgery treatment of laryngeal carcinoma T1]. AB - The early stage cancer of the glottis, including Tis, T1a and T1b stages, are the most common forms with the incidence rates ranging from 25% to 85%. The therapy of early glottic cancer is usually successful for two reasons. First, true glottic cancer produces early symptoms and it is relatively easy to remove. Second, glottis is rather poor with lymph pathways so the regional metastases are rare, less than 1%. Due to role of the larynx in phonation, respiration and swallowing, the cancer of this region and its treatment has a great impact to the quality of life. Retrospective study involved ten-year period, from January 1990 to January 2004. At the Institute for Otorhinolaryngology and Maxillofacial Surgery, Clinical Center of Serbia, 858 surgical and 54 endoscopic CO2 laser assisted resections were performed for glottic cancers of larynx of Tis-T2 stages. Glottic tumors were treated by Types III, IV and Va chordectomies according to classificaiton of endoscopic chordectomy defined by the European Laryngological Society. Analyzing the operated patients, as well as the type of the applied surgery, that is, endoscopic-laser and classic surgery, the authors attempted to clarify the dilemmas relating to the indications for one or another type of surgical intervention. The patients who had undergone primary radiotherapy were excluded from the analysis. PMID- 15756783 TI - [Results of hemilaryngectomy in primary surgical treatment of laryngeal carcinoma]. AB - Hemilaryngectomy is the resection of a true anatomic half of the larynx with preservation of the cricoid cartilage. We present a retrospective study of 438 patients with glottic carcinoma, treated with hemilaryngectomy, at the Institute of Otorhinolaryngology and Maxillofacial Surgery, Clinical Center of Serbia between 1988 and 1997. The patients with positive margins (19.4% of all) were postoperatively irradiated. Local recurrences of carcinoma were found in 17.3% of subjects, and regional recurrences in 16.4% of subjects. Those patients were treated with total laryngectomy or radical neck dissection, and with radiotherapy. 5-years survival rate in our patients was 79%. Hemilaryngectomy provided acceptable percent of local and regional recurrences, and good functional results: respiration, swallowing and voice quality. Therefore it could be the first choice surgery technique in treatment of T2 laryngeal carcinoma. PMID- 15756784 TI - [Radical surgery of the malignant laryngeal tumors]. AB - Modern therapeutical protocols for treatment of T3 and T4 malignomas of the larynx are not adjusted, because there are attempts to treat these diseases with non-operative methods (such as chemo- and radiotherapy) in order to preserve the organ. The aim of the study was to establish today's results of the surgical treatement of patients with T3 and T4 laryngeal malignoma. We studied the group of patients with laryngeal carcinoma, who had undergone total laryngectomy, during the period of eight years (1990-1997). The patients' data was submitted from medical documentation, it was filled in specially designed questionnaries and was statistically reviewed. During this eight-year-period, 1054 total laryngectomies were done. The five-years survival rate, established in the group of patients who had undergone total laryngectomiy is 308/794 (39%). In the patient group where total laryngectomy was salvage surgery after radiotherapy, the five-years survival rate is 47/172 (27%). In the patient group where total laryngectomy was salvage surgery after conservative or reconstructive surgery, the five-years survival rate is 28/84 (33%). Despite diagnostical and therapeutical achievements, prognosis for T3 and T4 malygnoma of the larynx was not significantly approved in the last few decades. PMID- 15756786 TI - [The analysis of the quality of life in laryngectomized patients]. AB - Total laryngectomy, as extremely mutilating surgical intervention, results in drastic changes of the style and quality of life. The trauma of laryngectomy is huge, both to patients and their environment. The most pronounced changes of the quality of life of laryngectomized patients are seen at marital, familial, professional, business and communication aspects. Malignant disease and total laryngectomy significantly reduce working capacity, producing, besides professional, the economical difficulties. Psychological implications associated with total laryngectomy are the most severe, comprehensive ones and require multidisciplinary approach. The study analyzes the frequency and distribution of factors affecting the quality of life of laryngectomized patients (sex, age, psychic status, probable history of chronic diseases, impairment of hearing and social-family environment of patients). Significant improvement of the quality of life of laryngectomized patients is achieved by systemic, planned and multidisciplinary rehabilitation of patients as well as their immediate environment. The need for the association of laryngectomized patients is stressed. PMID- 15756787 TI - [Strategy in the treatment of hypopharyngeal and cervical esophageal carcinoma]. AB - In the period between January 1st, 1978 and January 1st, 2003, 82 unselected patients with hypopharyngeal and cervical esophageal squamocellular carcinoma were treated at the Department of Esophagogastric Surgery, First University Surgical Hospital, Clinical Center of Serbia. In 43 (52.4%) patients operated with curative intent, radical surgical en-block resection and functional neck dissection has been performed. In 26 (60.5%) patient reconstruction was performed with stomach, in 11 (25.6%) left colon, and 6 (14%) free jejunal transfer. The overall 2-year and 5-year survival rate were 55.88% (19 patients) and 26.47% (9 patients), respectively. No patient undergoing nutritive gastrostomy and radiotherapy was alive after two years. At present surgery looks like the treatment of choice for hypopharyngeal and cervical esophageal carcinoma, providing a definitive palliation of dysphagia and better long-term survival. Free jejunal transfer has become the standard technique for reconstruction of the pharynx and hypopharynx especially with proximal lesions, whereas, gastric tube interposition is the technique of choice for reconstruction of the hypopharynx and cervical esophagus when the resection extends below the thoracic inlet or when there is a presence of synchronous carcinoma of theesophageal. PMID- 15756785 TI - [Effect of the extents of surgical resection in total laryngectomy on esophageal voice and speech rehabilitation]. AB - The study of functional results of total laryngectomy cannot be separated from articulation of esophageal voice and speech. To achieve success of phoniatric rehabilitation in laringectomized patients, it is necessary to be well aware of pathological sequelae of the very operation, long-term adverse effect of oncological risk factors as well as postoperative anatomo-physiological changes. Anatomic results of total laryngectomy depend on the type of surgical intervention and probable radiotherapy. The extent of surgical procedure, primary dictated by oncological indications, has the essential effect to articulation of esophageal voice and speech. If possible for oncological reasons, hyoid bone should be particularly left intact. Extensive surgical interventions of hypopharynx and the base of the tongue during laryngectomy, neck dissection- especially the radical one, and subsequent radiotherapy, all of them significantly reduce the possibility to produce esophageal voice and speech. Total laryngectomy eliminates the creator of voice and vibrations, which are acoustically perceived as esophageal voice, proceeding at the level of pharyngoesophageal junction. For this reason, the quality of newly created vibrating narrowing is especially important. PMID- 15756788 TI - [Carcinoma of the skin of the external auditory meatus]. AB - Carcinoma of the external meatus is a rare disease and a challenge to treat. Demographic and clinical characteristics of this disease are discussed for 14 patients treated in the Institute of Otorhinolaryngonlogy and Maxillofacial Surgery during the period 1993-2003. using four different clasification of the disease. All patients were treated surgically with postoperative radiotherapy. Five years survival is 100% for the II and II stage of the disease for all clasifications, and 75% for the III stage according to Pitsburg clasiffication. Reccurence of the disease in the first year after treatment is 66.6% and, 84% in the second year. The diagnostic protocol is clinical examination, computed tomography of the temporal bone, nuclear magnetic resonance of the temporal bone and endocranium, ultrasound examination of the neck and parotid region and tumor biopsy. The surgical approach could be less radical, through mastoid, with postoperative PMID- 15756789 TI - [Clinical study of undifferentiated carcinoma of the nasopharynx (UNCT) in patients with cervical node type of disease]. AB - Indifferentiated carcinoma of the nasopharyngs is clinicaly-histological imunologic entity which is often diagnosed in our country. There are three clinical types, but nodal cervical type of disease is the most interesting type for surgeons while the combined type is more interesting for otolaiyngologist. Among seventy-seven patients diagnosed with undifferentiated carcinoma of the nasopharyngs with nodal cervical type of disease, on the Institute of Otolaryngology and Maxillofacial Surgery Clinical Centre of Serbia during the period between 1993-1997 there were N0-21%, N1-49%, N2-18% i N3-12%, no mater of the T category. The disease more often occurs between male population (2:1), mostly between age 41-60. The rate for five year period of surviving for two different chemioterapeutical protocols is as follows: for categories N0 and N1 20% for mono Zorubicin and 61% for the same category for Z-CDDP. The same rate for categories N2 and N3 is 11% for mono Zorubicin and 33% for the same category for Z-CDDP. Much better rate of survival in comparison with previous decades is achieved due to better diagnosing on time in which are sistematicaly ineluded epypharyngoscopy in general anestesia with biopsy, CT and NMR and EBV serology. PMID- 15756790 TI - [Modern diagnostic procedures of nasopharyngeal carcinoma]. AB - The aim of this study is a research of single photon emission tomography (SPET) values with talium 201 and tehnetium 99m MIBI, comparing it with other methods CT, MRI and ultrasonography. In our investigation there were two groups of patients 24 treated with T1-201 and 17 with Tc-99m-MIBI. Before the therapy 7 patients had been tested with T1 201 and was 100% proved the sensibility, specificity, sensitivity and also 6 patients as really positive TP and 1 patient really negative TN. The same result was gotten in the group where TC 99m was used, where 3 of them were really positive and 1 negative. All discoveries were confirmed pathohistologically and leaning on this we determined the status of really positive and really negative patients. After the therapy there were 7 positive and 7 negative patients, and also one positive, and 2 negatives that were false, who were treated by thalium. On the basis of formula of sensibility these findings show that 77,8%, if it observed only a group of patients who were being tested after the therapy, while all sensibility of methode is 86.7%, where in the analise was included a group of patients at whom was registred the primary tumor. The specificity of the method for this group of patients after the therapy is 87.5%, and the whole 88.9% where the accuarsy is 82.4% in the group after therapy and the whole is 87.5%. PMID- 15756791 TI - [Occult metastases of oral cavity cancers]. AB - Despite new approaches to treatment and lower mortality, malignant tumors of the head and neck, including the malignant tumors of the oral cavity, still represent significant oncological problem because long-term survival has not been significantly prolonged. The growth of tumors of this localization is fast and infiltrative, while early metastases of regional lymph nodes are rather frequent. Malignant tumors of the oral cavity account for 1.1% of population in our community (Dimitrijvic, 2001). The objective of the study was to analyze regional metastases of the cancers of the tongue and the floor of mouth in 101 patients with planocellular cancers treated in the period 1991 to 1995. Clinically positive regional lymph nodes were found in 67.3% of patients, while the most commonly involved regions were submandibular (47.4%) and upper jugular region (46.1%). They were more frequent in localization of the floor of mouth than in case of tongue cancer. Three types of neck dissections were used for surgical treatment of patients. In the group of patients with clinically negative results of the neck (N0) who underwent neck dissection, occult metastases of regional lymph nodes were verified pathohistologically in 19.2% of the time. Malignant tumors of the oral cavity are always the indication for neck dissection, even in N0 category, on account of high proportion of occult metastases. PMID- 15756792 TI - [Distribution of cervical metastases from carcinoma of the cervicofacial region]. AB - The primary goal in the therapy of patients with cervico-facial cancers has been always the control of loco-regional disease. It is more difficult to control metastasis than primary tumor. According to numerous authors, metastases to cervical lymphonodus reduce the survival of patients with planocellular cancer of the upper aero-digestive pathways for about 50%. Precise classification of primary tumor and regional lymphonodus is highly significant for adequate and timely treatment of patients with cancers of cervico-facial region. The objective of our study was to make clinical classification of cervico-facial tumors and to establish the distribution of nodes according to node groups and cervical levels. In our series of 319 subjects, T2 category of primary tumors was most prevalent accounting for 40.44%. Clinically palpable lymphonodes were found in 87.15%, with most prevalent N1 category accounting for 42.95%. The incidence of clinically negative cervical nodus (N0) was reported in 12.85%. The nodes of the upper, medium and lower jugular group were most frequent in cancers of the larynx and pharynx. In cancer of the oral cavity, submental and submandibular nodes were the most commonly involved. In distribution of nodes based on oncological cervical levels, 45.86% of nodes at level II of the neck were found in laryngeal cancer, while 40% the neck level I was involved in the cancer of the oral cavity. In epipharyngeal cancer, 3.15% of cervical metastases were detected in the posterior triangle of the neck. The incidence of cervical metastases in specific primary localizations has a significant role for indications of one of the dissections of the neck. PMID- 15756793 TI - [Classification of chronic inflammation of the upper respiratory tract based on allergy status]. AB - Almost one third to one half of all patients in otorhinolaryngologic practice experience some kind of inflammation of the upper respiratory tract out of which allergic mechanisms, either as primary factors or secondary ones, appear in 30 40% of adults and 60-80% of children and adolescents. The objective of this study was to analyse inflammatory conditions of the upper airways on the basis of allergic state of the patient and to establish the classification that will respect the actual immunological alteration level (subclinical allergy, clinical allergy) and spreading (localized allergy, generalized allergy). Inclusion criteria for all sixty nine patients were the diagnosis of chronic upper airway inflammation and their exposition just to ubiquitous allergens. Diagnostic procedure included anamnesis, physical examination and allergic in vivo testing of the skin and nasal mucosa to inhalant allergens. The certain categories of results were established for the skin prick-test (positive, negative, indefinite), specific nasal provocation test (positive, negative, hyperreactive) and nasal symptoms (present, absent). By using a strictly determined combination of results, we were able to define the six groups in our classification: nasal clinical allergy (30% of patients), non-nasal clinical allergy (19% of patients), localized nasal allergy (11% of patients), latent allergy (3% of patient), nonspecific nasal hyperreactivity (12% of patient) and non-allergic inflammation (25% of patients). Our classification takes into consideration the modern knowledge in the field of allergology and may bring an additional quality in respect to selection of therapy options, long-term follow-up of allergy status evolution in the individual person as well as intragroup and intergroup analysis of parameters important to evaluate the effects of antiallergic prevention or therapy. PMID- 15756794 TI - [Iatrogenic perforations of the esophagus and hypopharynx--5 year experience at the Center for Esophageal Surgery]. AB - Iatrogenic perforations of the esophagus and hypopharynx are important problem, due to diagnostic difficulties, controversies about adequate treatment, and high morbidity and mortality rate. Incidence of iatrogenic perforations is from 50 to 75% of all perforations. In the period from April 1999. to April 2004, 15 patients with iatrogenic perforation of the esophagus and hypopharynx were treated at the Department of esophageal surgery, First University Surgical Hospital in Belgrade. In majority of patients iatrogenic perforation occured during endoscopic interventional procedure (endoscopic removal of ingested foreign body--10 pts, endotracheal intubation--2 pts, intraoperative iatrogenic perforation--2 pts, pneumatic dilatation--1 pt). Surgical treatment was performed in 12 (80%) pts and 3 (20%) pts were treated conservatively. Surgical approach was cervicoabdominal, thoracoabdominal and cervicothoracoabdominal in 9.1 and 2 pts, respectively. Among 12 operated pts, primary repair of the esophagus was performed in 5 pts, and esophageal resection or exclusion in 7 pts. Overall mortality rate was 13.3% (2 pts), in surgical group 8.3% (1 pt) and in conservatively treated group 33.3% (1 pt). Iatrogenic perforations of the esophagus and hypopharynx are diagnostic and therapeutic problem. Awareness of the possibility of esophageal perforation during instrumental manipulations and early diagnosis is essential for successful, individually adapted, and in most cases surgical, treatment. PMID- 15756795 TI - [Surgery of cholesteatoma: surgical technique and cholesteatoma recurrences]. AB - Past experience with open and closed techniques of tympanoplasty in surgery of cholesteatoma has shown that recurring illness is one of the major causes of surgical failure. The literature has reported varying trend of surgical treatment of cholesteatoma. The objective of the study was to analyze the significance of surgical technique in relation to the incidence and most frequent localization of recurrent cholesteatoma. Our study analyzed 120 patients operated on for cholesteatoma. The patients were divided into two groups, group I (45) with recurring disease and group II (75) without any recurring condition, which were followed up three years. Statistical analysis was carried out by modified t-test. The largest number of patients was re-operated in the first two years from the initial surgery (50%), In the majority of patients (50%), recurrent cholesteatoma was most commonly localized (stage I) in attic (20%) and much rarely in mesotympanum (11.9%). Stage III recurrent cholesteatoma was verified in 35% of patients, most frequently diffuse form (13.4%). The involvement of attic by all three stages of disease accounted for over 60%. The analysis of the used techniques of surgical treatment in both groups revealed significant difference. Open techniques of tympanoplasty were used in 60% of patients with no recurrence. Closed techniques were used more frequently in patients with recurring disease, i.e. in over 90% of cases. Recurerrent cholesteatoma develops, in the majority of cases, during the first two years after the surgical intervention. Attic is the most common localization of cholesteatoma. More frequent utilization of open technique of tympanoplasty for surgery of cholesteatoma significantly reduces the incidence of recurring condition. The indications for CWD technique are the initial spread of cholesteatoma, possibility of complete removal of cholesteatoma and postoperative follow-up of patients. PMID- 15756796 TI - [Mastoiditis in adults: diagnostic and therapeutic aspects]. AB - The purpose of the present study is to report our experience in the diagnosis and treatment of mastoiditis in adults. Five patients with an active chronic otitis and mastoiditis were presented. All of them had a history of chronic ear discharge for long period of time and have been diagnozed and treated sufficiently. All relevant data were analyzed from the medical records. The most common symptoms of the disease were otalgia, otorrhea and hearing loss, but the physical signs of mastoiditis (swelling, erithema and tenderness of the retroauricular region) were presented in all cases. Localization and enlargement of the pathological process within the middle ear spaces was determined by CT. All patients were treated surgically and with intravenous antibiotics. In one patient the other treatment was applied due to a specific (TBC) process in the ear. Early diagnosis and adequate treatment (surgery combined with an effective antibiotics therapy) is most important to prevent a serious complications of mastoiditis (extracranial and/or intracranial). PMID- 15756798 TI - [Radiologic evaluation of malignant diseases of the larynx]. PMID- 15756797 TI - [Clinical study of routes of onset of inflammatory diseases in the maxillary sinuses]. AB - Pathways of the maxillary sinus diseases is an interesting issue investigated by many authors during the past decades. The goal of this research was to study the pathways of infection spreading into the maxillary sinuses and to compare them in relation to their frequency, underlying causes and general epidemiologic characteristics of the involved population. A total number of 150 adult patients of both gender suffering different diseases of maxillary sinuses were included into the study. Pathways rising maxillary sinuses diseases were diagnosed on the basis of standard clinical procedures including CT scean and MRI of the region. We found inflammatory processes to dominate the tumorous ones (107:43 patients). Rhinogenic type of sinusitis was the most frequent disease (72 patients) while odontogenic sinusitis (35 patients) was significantly less frequent. No case of traumatic or hematogenic maxillary sinusitis was found. Rhinogenic maxillary sinusitis is characterised by spontaneous onset while odontogenic one is mostly of arteficial origin after surgical procedures in the oral cavity (88% of patients). In contrast to rhinogenic type, odontogenic maxillary sinusitis is far more frequent in younger patients. PMID- 15756799 TI - [Gaucher disease: diagnosis and treatment]. AB - Gaucher's disease is the most common lysosomal storage disorder. It was identified in 1882 by Phillipe Gaucher, a French dermatologist. However, it was not until 1965 that Gaucher disease was found to be due to a deficiency in the enzyme glucocerebrosidase (EC 3.2.1.45) which breaks down glucocerebroside, a cell membrane component. The deficiency in this enzyme leads to an accumulation of glucocerebroside within the lysosomes of macrophages throughout the body. Gaucher's disease is classified into three types: type 1 (non-neuronopathic), type 2 (acute neuronopathic), and type 3 (subacute neuronopathic). Of the three, type 1 is the most common, affecting one in 40,000-200,000 people and having a high prevalence among Ashkenazi Jews, affecting one in 450-1500. The signs and symptoms of type 1 disease demonstrate marked heterogeneity, from asymptomatic or mildly symptomatic, to severe disability with disfigurement and even death. Hepatosplenomegaly and thrombocytopenia are well documented. Less well-recognized are often insidious skeletal complications which affect the majority of type 1 patients and which are its most debilitating feature. In addition to clinical suspicion, some morphologic, hematologic and biochemical indicators can help establish the diagnosis. However, definitive diagnosis is only made by determining the catalytic activity of the lysosomal enzyme glucocerebrosidase. Confirmation of heterozygosity requires the use of molecular biotechnology methods. About 150 mutations of the glucocerebrosidase gene have been identified in patients with Gaucher's disease, some of which are predictive of phenotype. The history of treatment of Gaucher disease started with splenectomy and continued with bone marrow transplantation, before the recent introduction of safe and effective enzyme replacement therapy. In Croatia, nine patients with type 1 Gaucher's disease have been identified so far. Seven patients are on enzyme replacement therapy, and past results demonstrated significant improvement in all clinical symptoms, without development of any side effects. However, new treatments, such as substrate balance therapy and gene therapy, may become available within the next few years. The place, if any, that such therapies will have in the treatment of patients with Gaucher's disease will be dependent on the results of clinical studies currently in progress. PMID- 15756800 TI - [Reference values of ventilatory capacity in persons in the third stage of life]. AB - Ventilatory capacity was measured in a group of 622 subjects aged 60 years and older (475 women and 147 men). METHODS: Maximum expiratory flow-volume curve was recorded, on which forced vital capacity (FVC), one second forced expiratory volume (FEV1) and maximum expiratory flow rates at 50% and the last 25% of vital capacity (FEF50, FEF25) were read. Reference values of ventilatory capacity tests for women and men were calculated by linear multiple regression with age and height as predictors, separately for smokers and nonsmokers. RESULTS AND CONCLUSION: Results of ventilatory capacity tests in our population showed better compliance with the reference values proposed by Mustajbegovic et al. compared to those by Quanjer et al. According to our results the ventilatory capacity values decreased by the age of 70-79 years, followed unexpectedly by an increase relative to the expected values. Since ventilatory capacity tests do not follow a linear decrease, in the evaluation of lung function in the elderly it is necessary to use reference values specific for this population. PMID- 15756801 TI - [Glycemic control and the risk of preeclampsia in women with gestational diabetes mellitus]. AB - INTRODUCTION: Gestational diabetes mellitus (GDM) is associated with an increased risk of preeclampsia and other complications throughout pregnancy. AIM: The aim of the study was to analyze the incidence of preeclampsia and other risk factors in GDM pregnant women. STUDY DESIGN AND METHODS: During 1992-2001 period, 472 consecutive unselected pregnancies were followed up in women with GDM. Glycemic control was assessed by HbA1c at the time of diagnosis. Preeclampsia was defined as blood pressure 140/90 mm Hg combined with albuminuria of >0.3 g/L. The occurrence of preeclampsia was also assessed in a control group comprising 324 unselected pregnant women. RESULTS: Preeclampsia developed in 14.4% of GDM women and 4.3% of controls. After adjustment by logistic regression, both fasting blood glucose (FBG) and profile blood glucose (PBG) and their changes during pregnancy remained significant predictors for preeclampsia. The odds for preeclampsia increased by a factor 1.2 for each 1 mmol/L increment in initial FBG level and PBG level, and decreased by factor 0.8 for each 1 mmol/L decrease of FBG or PBG level achieved during pregnancy. CONCLUSION: Improvement of the glycemic control during pregnancy reduces the risk of preeclampsia. PMID- 15756802 TI - [Antioxidative activity of propolis from Dalmatia (Croatia)]. AB - AIM: The aim of this study was to determine the antioxidative activity of propolis from ecologically clean parts of Dalmatia. METHODS: Phenol concentration in ethanolic propolis extracts was determined by Folin-Ciocalteu reagent using gallic acid as the standard. Flavonoid phenolic compounds were analyzed after precipitation with formaldehyde. The residual non-flavonoid phenolics were also determined by Folin-Ciocalteu method. By determining the change of peroxide number (PN), of tiobarbiture acid reactive species (TBARS), and of DPPH-radical activity, antioxidative efficiency of propolis was tested and compared with well known and widely used synthetic antioxidants. Values of PN and TBARS were determined at 60 degrees C in samples of trigyceride substrate (lard) without and with the addition of antioxidants. Compared was the efficiency of three antioxidants: propolis (alcoholic extract), vitamin E, and (+)-catechin in a concentration of 1%. PN was monitored during 50 days. By the method of Sedlacek, TBARS were measured during 30 days. Antioxidative activity of propolis extract was also measured in terms of hydrogen donating ability using stable radical alpha,alpha-diphenyl-beta-picril hidrazyl (DPPH*) and compared with commercial synthetic antioxidants of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and (+)-cathecin. Inhibition degree of DPPH* was calculated by the formula of Yen and Duh. RESULTS: Total phenol content, expressed as gallic acid, in propolis extracts varied from 75.2 to 90.2 g/kg propolis. The proportion of flavonoids in total phenols ranged from 62% to 65%. Values of TBARS were not increased only in samples with added propolis. The inhibition of DPPH-radical by propolis extracts ranged from 93% to 96%, by catechin 95%, by BHT 49%, and by BHA 64%. Compared to BHT and BHA, propolis extracts showed greater reducing activity against DPPH-radical. DISCUSSION: The chemical composition of propolis, and thus its biological activity depend on the plant from which it has been collected, and on the macro- and microclimatic conditions. Many compounds in propolis exert antioxidative activity. A belief was expressed that the biological activity of propolis is very probably based mostly on its antioxidative efficiency. Dalmatian propolis showed high efficiency in the prevention of oxidative processes. This could be explained by the high proportion of polyphenol constituents, especially flavonoids. A very low and equal degree of increase of PN, as a measure of oxidative processes, was noticed in the samples of triglyceride substrate with the addition of propolis and (+)-catechin. The greatest rise of TBARS was measured in the samples of pure lard. There was no increase of TBARS only in the samples with added propolis. Propolis and (+)-catechin showed great efficiency in the inhibition of DPPH-radical, greater than BHT and BHA, which are widely used in food industry. CONCLUSION: The results indicate that Dalmatian propolis could be an efficient protective agent against oxidative processes in food. The high antioxidative activity of propolis, its natural origin, and present knowledge about its biological properties, make it a very promising nutritional additive for human diet. PMID- 15756803 TI - [Clinical characteristics of pruritus in hemodialysis patients]. AB - AIM: Aim of the research was to analyze clinical characteristics and most important risk factors of uremic pruritus. PATIENTS AND METHODS: A total of 151 patients on chronic hemodialysis (CHD) during at least 12 months were analyzed. Thorough history was taken for pruritus, its presence and localization, sleep disorder and neuropathic symptoms. On physical examination, attention was focused on the skin. Laboratory tests includes blood cells count, serum urea, creatinine, electrolytes, aminotransferases, alkaline phosphatase and proteins. The dose of dialysis was followed by Kt/V. On statistical analysis, t-test and chi2 test were used. RESULTS: Pruritus was present in 85 (56%) patients, 41 women and 44 men, mean age 53.56+/-13.36 (26-81) years, mean time on CHD 78.36+/-55.02 (12-268) months. There were 66 (44%) patients without pruritus, 32 women and 34 men, mean age 50.35+/-13.76 (22-73) years, on CHD for 58.64+/-50.40 (12-187) months. Although the patients with pruritus were somewhat older and longer on CHD, there was no significant difference either in sex structure or distribution according to primary renal disease. In the group with pruritus there were significantly more anuric patients (43 vs. 22) (p<0.01). The patients with pruritus had a higher rate of sleep disorder (NS), calcium deposits in soft tissues and blood vessels (NS) and clinical neuropathy (p<0.01). Skin changes were found in almost all patients with pruritus (93%), which differed significantly from the patients without pruritus (48%) (p<0.005). The mean value of Kt/V was 1.23+/-0.35 in patients with pruritus, and 1.34+/-0.41 in those without pruritus (NS). Kt/V higher than 1.4 was significantly less frequently recorded in patients with pruritus than in those without pruritus (55%) (p<0.005). The red cell and white cells count, serum hemoglobin, calcium, phosphorus and their products, aminotransferases, bilirubin, alkaline phosphatase and proteins were approximately the same in both groups of patients. CONCLUSION: The loss of residual renal diuresis, Kt/V below 1.4, presence of calcium deposits and neuropathy were the most common risk factors for the extent of uremic pruritus in our CHD patients. PMID- 15756804 TI - [New biochemical markers in the assessment of minor myocardial damage in critically ill patients]. AB - The diagnosis of acute myocardial infarction (AMI) and unstable angina is based on typical clinical signs, electrocardiogram (ECG) changes, and serial measurements of characteristic serum enzymes, especially troponins. Today, newer biochemical markers are used in assessing these conditions, as well as minor myocardial damage (MMD). AIM: The aim of this study was to determine the existence of MMD by the analysis of new biochemical markers including cardiac troponin T (c-TnT), troponin I (c-TnI), CK-MB activity (CK-MBact), CK-MB mass (CK MBmass), a conventional marker of total creatine kinase (CK), and 12-lead ECG monitoring in two groups of critically ill patients admitted to the intensive care unit (ICU). Group 1 (n=52) consisted of the patients with heart failure, unexplainable hypotension, chronic obstructive pulmonary disease in acute exacerbation, acute severe pancreatitis, sepsis, pulmonary embolism, diabetes with complications, liver cirrhosis, and tachycardia >120/min who suffered chest discomfort without evident ECG signs of AMI. Group 2 consisted of patients (n=73) with acute gastrointestinal bleeding, poisoning, and miscellaneous conditions without chest discomfort or ECG signs of AMI. RESULTS: Of the 52 critically ill patients, 21.2% were positive for troponin T and 11.5% for troponin I. Group 1 patients showed a prevalence of elevated total CK (28.8%), CK-MBact (7.7%), and CK-MBmass (32.7%). In group 2, positive troponin I was found in 19.7% of patients, troponin T was negative (0.00%), with elevated total CK (23.9%), CKMBact (7.0%), and CK-MBmass (2.3%). The mean concentrations of c-TnT, c-TnI, total CK, CK-MBact, CK-MBmass were higher in group 1 than in group 2, however, without statistical significance. The best positive correlation was recorded between CK-MBact and CK-MBmass (r=0.63). The c-TnI showed best discrimination and accuracy in the assessment of MMD under the ROC curves surface (0.84), while the accuracy was low for all other markers analyzed. Discrimination and accuracy were moderate for all markers analyzed. DISCUSSION AND CONCLUSION: CK-MBact (92.3%) showed the highest specificity, followed by c-TnI (88.5%) and c-TnT (75.6%). The sensitivity was low for all markers analyzed. Concerning specificity, CK-MBact proved to be the best biologic marker for the assessment of MMD, followed by c TnI and c-TnT. Correct clinical assessment according to marker accuracy and discrimination can be achieved by use of c-TnI, however, with a moderate degree of accuracy and discrimination for the detection of MMD in critically ill patients admitted to ICU. PMID- 15756805 TI - [Coagulation tests in septic surgical patients]. AB - AIM: To determine whether sequential change in coagulation parameters such as activated partial thromboplastin time (aPTT), prothrombin time (PT), platelets count and fibrinogen level may predict the outcome of patients in sepsis. STUDY DESIGN: Cohort longitudinal study. PATIENTS AND METHODS: Patients with positive two or more clinical criteria for sepsis were eligible for the study. Thirty patients were included, 24 male and 6 female. Eight patients survived, while 22 deceased. Median age of survivors was 66 years (range 23-77), and in non survivors it was 69 years (range 48-79), p=0.37. In 9 patients malignancy was an underlying disease. APACHE II score was calculated at admittance, median value for survivors was 10 (range 7-15) and for non-survivors it was 26 (range 6-35), p=0.001. Calculated MODS score at the time blood cultures was 2 (range 0-9) for survivor and 6.5 (range 2-13) for non-survivors, p=0.007. Blood cultures were taken at the onset of sepsis, and in 29 patients they were positive. Coagulation parameters were measured at admittance, at the onset of sepsis and 48 hours after the introduction of the specific antimicrobial therapy. RESULTS: Analysis of variance for repeated measurements between survivors and non-survivors has shown that there were no differences in values of coagulation parameters. The only significant difference between these groups of patients was APACHE II and MODS score. In 7 patients with severe thrombocytopenia (<33,000 x 10(9)/L) as a result of irreversible septic shock a clinically visible bleeding was present in only one patient. DISCUSSION: Disseminated intravascular coagulation (DIC) is a clinical-pathological syndrome in which wide-spread intravascular coagulation is induced by procoagulants that are introduced or produced in the blood circulation and overcome the natural anticoagulant mechanisms. DIC causes tissue ischemia from occlusive microthrombi as well as bleeding from both the consumption of platelets and coagulation factors and the anticoagulant effect of products of secondary fibrinolysis. In sepsis, tissue factor which is the most common trigger of DIC can be generated and expressed on membranes of monocytes and endothelial cells during the systemic inflammatory response syndrome (SIRS). The wide-spread generation of thrombi in sepsis induces deposition of fibrin which leads to vessels obstruction and consumption of substantial amounts of haemostatic factors i.e. platelets, fibrinogen, factors V, VIII and others, protein C and antithrombin III (AT III). Intravascular thrombi trigger secretion of tissue plasmin activator (tPA) from endothelial cells which sets of compensatory thrombolysis which may reopen the occluded blood vessels. But byproducts of thrombolysis such as fibrin/fibrinogen degradation products may enhance bleeding by interfering with platelet aggregation, fibrin polymerization and thrombin activity. The typical feature of sepsis is depression of three powerful anticoagulant systems: protein C pathway, AT III pathway and tissue pathway factor inhibitor (TPFI). This sequence of events led us to hypothesize that alterations in coagulation parameters such as PT, aPTT, fibrinogen, platelets count may predict the outcome of disease, as it is well documented that the development of DIC confers prognosis of sepsis. The failure to distinguish survivors from non-survivors by the alteration in the coagulation parameters in this study may be due to a relatively low sample size or to the clinical necessity of an attending physician to substitute the deficient blood or coagulation product. CONCLUSION: The coagulation parameters PT, aPTT, platelet count and fibrinogen level can not serve as predictors of outcome in patients with sepsis. Further studies including more discerning coagulation parameters: AT III, D-dimer, soluble fibrin monomer, thrombin/antithrombin complex, plasmin/antiplasmin complex, fibrinopeptid A, fibrinopeptid B are necessary to evaluate whether these procoagulant and anticoagulant factors may help in predicting outcome and severity of sepsis. PMID- 15756807 TI - [Problems of erectile dysfunction prevalence]. AB - According to non-clinical studies, erectile dysfunction (ED) seems to be the fastest growing among male sexual problems. Its prevalence range widened from 4% 9% in 1990 to 10%-52% in 2000. The paper is an attempt at understanding this new and rampant "epidemic". Instead of trying to pinpoint its "natural" causes, the author aims to decipher the socially constructed dynamics of the disorder. The approach is based on a qualitative investigation of the scientific conceptualization and measuring of ED. The analysis points out serious shortcomings, which contradict the much-celebrated usefulness of the 1992 NIH Panel's consensus on impotence. More importantly, the observed measurement differences call into question the usefulness of the current research on the prevalence of ED. Using data from the first Croatian study on male sexual dysfunction, the author demonstrates the seriousness of problems stemming from the use of non-standardized measures. In conclusion, the steps for reaching the necessary standardization are outlined. PMID- 15756806 TI - [Interleukin-2 receptor blockers in renal transplantation]. AB - The last two decades have witnessed significant advances in the renal transplantation immunosuppressive protocols. The introduction of mycophenolate mofetil, tacrolimus, sirolimus and polyclonal antibodies has significantly improved graft survival. However, intensification of immunosuppression results in complications such as malignant diseases, opportunistic infections and metabolic disturbances with consequential increase in cardiovascular mortality. Advances in molecular engineering have made possible the development of monoclonal humanized or chimeric antibodies, which will not induce the host immune response with production of neutralizing antibodies or serum sickness. Antibodies directed against the alpha chain of human IL-2 receptor have recently been introduced into immunosuppressive protocols. Daclizumab is a humanized antibody, and basiliximab is a chimeric antibody, engineered by cloning segments of the murine immunoglobulin sequence into the human-immunoglobulin gene. It decreases immunogenicity while maintaining high specificity for IL-2R alpha chain. The efficacy and safety of both preparations have been reported in large randomized studies. Their use in induction resulted in a significant decrease in acute graft rejections after renal transplantation. The possibility of decreasing the dose or complete withdrawal of certain immunosuppressive agents with the use of IL-2R blockers seems promising for further improvement in the longterm graft survival. Longterm follow-up is necessary to determine their role in solid organ transplantation. PMID- 15756808 TI - [Lid retraction etiology]. AB - The upper lid position is abnormal if it exposes a white band of sclera between the lid margin and the upper corneal limbus while the retracted lower lid lies below the inferior corneal margin and is tethered to the orbital margin. Lid retraction is a sign of many congenital and acquired diseases and is characterised by multifactorial etiology. The aim of this study was to discuss the etiology of lid retraction divided into four categories: neurogenic, myogenic, mechanical and miscellaneous, what suggests a successful differential diagnostic and therapeutic approach. PMID- 15756809 TI - [Familial adenomatous polyposis with extracolonic manifestations--case report]. AB - The familial adenomatous polyposis syndrome is an autosomal dominant inherited disease characterized by progressive development of multiple adenomatous polyps throughout the colon and rectum. Due to the malignant potential of adenomatous polyps, colorectal cancer develops in 100% of cases, approximately 10-15 years after the onset of symptoms. Extracolonic manifestations of the disease including adenomatous polyps of the stomach, duodenum, small intestine and periampullatory region are rare. The etiology of the disease is germline mutation at the site of tumor suppressor gene located on chromosomes 5q21-22. A case is described of a 48 year-old man hospitalized at the Department of Abdominal Surgery, Sveti Duh General Hospital in Zagreb for the treatment of familial adenomatous polyposis syndrome. For some time the patient reported occasional abdominal pain, frequent stools and diarrhea with blood, anemia and body weight loss. Laboratory, radiology and endoscopy examinations verified multiple adenomatous polyps of the colon and rectum, also with polyps of the stomach, duodenum and jejunum. Histopathology confirmed the polyps to show moderately poorly differentiated cylindric epithelium and moderate to severe dysplasia. Radical surgery was required, so proctocolectomy with Brook ileostomy was performed. The postoperative recovery and wound healing were normal. The patient was discharged twelve days of the surgery for home care. Oncologic treatment was suggested. Verified extracolonic manifestations of the disease require periodical endoscopic follow up and possible treatment. PMID- 15756810 TI - [Acute renal failure in patients undergoing cardiac surgery]. AB - Acute renal failure (ARF) is an unusual and severe complication which may occur in patients following cardiac surgery. The incidence of ARF is from 1% to 15% (according to some authors up to 40%). The ARF, occurring in the postoperative period and requiring dialysis is an important risk factor for early mortality, while the overall mortality due to this complication is as high as 40% (40% to 90%). AIM: The aim of this study was to assess the incidence of ARF in patients undergoing cardiac surgery at our hospital from January 1, 2001 to June 1, 2002 and to compare it with the data obtained at the same institution and published 17 years ago. METHODS: A total of 290 patients undergoing cardiac surgery were analyzed, 71 (24.5%) female and 219 (75.5%) male, mean age 61.1 (range 17-81) years. Exclusion criteria were death within a few hours of surgery and need of chronic hemodialysis prior to surgery. ARF was defined as doubling of serum creatinine (sCr) concentration with preoperative sCr concentration below 130 micromol/L, or sCr increase by 100 or more micromol/L after cardiac surgery. Age, sex, type of surgery, preoperative sCr and preoperative risk factors (hypertension, diabetes mellitus, hyperproteinemia, pulmonary disease, peripheral vascular disease, central vascular disease) as well as complications occurring during the operation and their influence on ARF were analyzed. The incidence of ARF, therapy and mortality were also analyzed. RESULTS: Ischemic cardiac disease was present in 236 (81%) and valvular disease in 41 (14%), ventricular or atrial septal defect in 6 (2%), thoracic aortic aneurysm in 3 (1%), patent ductus arteriosus in 2 (0.7%) patients, and pericardial tumor and penetrant pericardial injury in 1 (0.36%) patient each. Arterial hypertension was present in 199 (68.6%), hyperlipoproteinemia in 194 (66.8%), diabetes mellitus in 76 (26.2%), cardiac arrhythmias in 39 (13.45%), cerebrovascular diseases in 32 (11.0%) previous, renal diseases in 25 (8.6%), chronic obstructive lung disease in 23 (7.9%) patients, peripheral vascular disease by 19 (6.6%) patients, thyroid disease by 8 (3.1%), and malignant disease in 5 (7.1%) patients. Renal function according to sCr was as follows: <79 micromol/L in 90 (31.0%), 80 to 99 micromol/L in 124 (42.7%), and 100-129 micromol/L in 58 (20%), 130-159 in 10 (3.4%), and >160 micromol/L in 4 (1.4%) patients. ARF developed in 8 (2.1%) patients who had undergone cardiac surgery. Among them, only one (0.3%) patient needed dialysis treatment (hemodialysis and continuous venovenous hemofiltration). There were no differences in sex distribution between the patients who developed ARF and those who did not. The patients who developed ARF were older, mean age 65.7 years. Most of the patients with ARF suffered from hypertension, diabetes mellitus and hyperlipoproteinemia. Seventy-five percent of patients who developed ARF had some kind of "surgical" complications: postoperative bleeding with developing hemorrhagic shock, myocardial infarction during the operation, or acute abdomen after the operation. CONCLUSION: The incidence of ARF in patients undergoing cardiac surgery was low (2.1%). The incidence of severe ARF (which must be treated with dialysis) was 0.3%. We compared the data obtained at our hospital with those obtained 17 years ago and found a reduction in the incidence of severe ARF after cardiac surgery (0.3% vs. 4%). PMID- 15756811 TI - [Some epidemiologic characteristics of foodborne intoxications in Croatia during the 1992-2001 period]. AB - Epidemiologic characteristics of foodborne intoxications registered in Croatia during the 1992-2001 period are described. Salmonellosis, as well as staphylococcal food intoxication, foodborne botulism, Clostridium perfringens food intoxication, histamine poisoning and food intoxication caused by other pathogens, especially conditionally pathogenic bacteria, were analyzed. During the study period, 87,782 cases were reported, 45,721 (52.1%) of them related to salmonellosis and the rest of 42,061 (47.9%) cases to foodborne intoxications of various etiology. The diseases occurred throughout the year, with a higher proportion in summer than at any other time of year. During the study period, 480 outbreaks of foodborne intoxications with 10,567 cases were reported, 405 (84.4%) of them with 8476 cases related to salmonellosis. The most frequently isolated pathogen in salmonellosis outbreaks was Salmonella enteritidis with 345 (85.2%) outbreaks. The outbreaks of salmonellosis were primarily associated with the intake of pastries (40.1%). Ice-cream was associated with a high risk of staphylococcal food intoxication, and bean salad of Clostridium perfringens food intoxication. The food vehicle of transmission most often associated with foodborne botulism outbreaks was smoked ham. Histamine poisoning, recorded in four outbreaks, occurred after consumption of fish and fish products. Salmonellosis outbreaks most frequently occurred at home (n=149; 36.8%), followed by pastry shops with 47 (11.6%) outbreaks, the latter also ranking first in the outbreaks of staphylococcal food intoxication (40%). PMID- 15756812 TI - Involvement of porin N,N-dicyclohexylcarbodiimide-reactive domain in hexokinase binding to the outer mitochondrial membrane. AB - The proportion of hexokinase that is bound to the outer mitochondrial membrane is tissue specific and metabolically regulated. This study examined the role of the N,N-dicyclohexylcarbodiimide-binding domain of mitochondrial porin in binding to hexokinase 1. Selective proteolytic cleavage of porin protein was performed and peptides were assayed for their, effect on hexokinase I binding to isolated mitochondria. Specificity of DCCD-reactive domain binding to hexokinase I was demonstrated by competition of the peptides for porin binding sites on hexokinase as well as by blockage hexokinase binding by N,N-dicyclohexylcarbodiimide. One of the peptides, designated as 5 kDa (the smallest of the porin peptides, which contains a DCCD-reactive site), totally blocked binding of the enzyme to the mitochondrial membrane, and significantly enhanced the release of the mitochondrially bound enzyme. These experiments demonstrate that there exists a direct and specific interaction between the DCCD-reactive domain of VDAC and hexokinase I. The peptides were further characterized with respect to their effects on certain functional properties of hexokinase I. None had any detectable effect on catalytic properties, including inhibition by glucose 6-phosphate. To evaluate further the outer mitochondrial membrane's role in the hexokinase binding, insertion of VDAC was examined using isolated rat mitochondria. Preincubation of mitochondria with purified porin strongly increases hexokinase I binding to rat liver mitochondria. Collectively, the results imply that the high hexokinase-binding capability of porin-enriched mitochondria was due to a quantitative difference in binding sites. PMID- 15756814 TI - Cloning and characterization of the xyn11A gene from Lentinula edodes. AB - Hemicellulose represents a rich source of biomass that can be converted into useful chemical feedstocks. One of the main components of hemicellulose is xylan, a polymer of xylose residues. Xylanase enzymes that hydrolyze xylan are therefore of great commercial interest. We have cloned a gene (xyn11A) that encodes a 283 amino acid xylanase enzyme from the fungus Lentinula edodes. The enzyme has a pI of 4.6 and belongs to the highly conserved glycosyl hydrolase family 11. The xylanase gene was cloned into a Pichia pastoris expression vector that secretes active enzyme into both solid and liquid media. The optimal reaction conditions were at pH 4.5 and 50 degrees C. The enzyme had a Km of 1.5 mg/ml and a Vmax of 2.1 mmol/min/mg. Xyn11A produced primarily xylobiose, xylotriose, and xylotetraose from a birchwood xylan substrate. This is the first report on the cloning of a hemicellulase gene from L. edodes. PMID- 15756813 TI - Structure-function relationship of new crotamine isoform from the Crotalus durissus cascavella. AB - In this work we isolated a novel crotamine like protein from the Crotalus durissus cascavella venom by combination of molecular exclusion and analytical reverse phase HPLC. Its primary structure was:YKRCHKKGGHCFPKEKICLPPSSDLGKMDCRWKRK CCKKGS GK. This protein showed a molecular mass of 4892.89 Da that was determined by Matrix Assisted Laser Desorption Ionization Time-of-flight (MALDI-TOF) mass spectrometry. The approximately pI value of this protein was determined in 9.9 by two-dimensional electrophoresis. This crotamine-like protein isolated here and that named as Cro 2 produced skeletal muscle spasm and spastic paralysis in mice similarly to other crotamines like proteins. Cro 2 did not modify the insulin secretion at low glucose concentration (2.8 and 5.6 mM), but at high glucose concentration (16.7 mM) we observed an insulin secretion increasing of 2.7-3.0 fold than to control. The Na+ channel antagonist tetrodoxin (6 mM) decreased glucose and Cro 2-induced insulin secretion. These results suggested that Na+ channel are involved in the insulin secretion. In this article, we also purified some peptide fragment from the treatment of reduced and carboxymethylated Cro 2 (RC-Cro 2) with cyanogen bromide and protease V8 from Staphylococcus aureus. The isolated pancreatic beta-cells were then treated with peptides only at high glucose concentration (16.7 mM), in this condition only two peptides induced insulin secretion. The amino acid sequence homology analysis of the whole crotamine as well as the biologically-active peptide allowed determining the consensus region of the biologically-active crotamine responsible for insulin secretion was KGGHCFPKE and DCRWKWKCCKKGSG. PMID- 15756815 TI - Structural changes of beta-lactoglobulin during thermal unfolding and refolding- an FT-IR and circular dichroism study. AB - We have quantitatively characterized by FT-IR spectroscopy the contents of secondary structure of beta-lactoglobulin during thermal unfolding and subsequent refolding. Our data clearly indicate that considerable amount of secondary structure, particularly beta-sheet, still remained intact even at 90 degrees C. Noticeable changes in secondary structure of beta-lactoglobulin were observed only above 70 degrees C. The refolded protein regained, within limits of experimental error, all of the secondary structure lost during thermal unfolding. The data also indicate that the refolding mechanism operating at pH 7.0 and 2.0 are the same. Identical secondary structure of native and refolded beta lactoglobulin was also indicated by far-UV circular dichroic spectra of the two forms of protein. Near UV circular dichroic spectra of the same two forms showed considerable differences indicating less tertiary structure of refolded beta lactoglobulin. The combined CD and FT-IR data indicated that refolded form of beta-lactoglobulin could be characterized as a molten globule state as it had native-like secondary structure and compromised tertiary structure. PMID- 15756816 TI - Oxygen-binding heme complexes of peptides designed to mimic the heme environment of myoglobin and hemoglobin. AB - Development of effective resuscitation agents for blood-loss replacement in trauma or surgery is extremely important despite substantial improvements in screening methods of blood from human donors. This paper reports the design and synthesis of peptides that mimic the natural environment of the heme group in myoglobin (Mb) and in the alpha- and beta-subunits of human adult hemoglobin (Hb). The designs were based on the fact that the heme group in the aforementioned proteins is sandwiched between helices E and F. Fifteen test peptides and six control peptides were synthesized, and their ability to form stable complexes with heme was investigated. It was found that none of the control peptides or proteins was able to bind heme. However, each of the peptides that were designed to mimic the E-F helices, and even shorter designs, which removed from this region residues that do not contribute to contacts with the heme group, were each able to bind one mole of heme per mole of peptide forming peptide heme complexes that were stable to manipulation and behaved as single molecular species. Oxygen binding measurements on the reduced peptide-heme complexes showed that these compounds bind oxygen and give visible spectra that were typical of oxygenated heme-proteins. In oxygen binding measurements done under different partial pressures of oxygen, the heme peptide complexes gave hyperbolic oxygen-saturation curves, but showed slight differences in their P50 values. The P50 values ranged from 3.8 mmHg for the heme peptide B7 complex to 13.7 mmHg for the heme peptide D13 complex (under the same conditions, P50 values for Hb and Mb were 34.0 and 5.5 mmHg, respectively). It is concluded that peptide constructs designed to mimic the heme-binding regions of Mb or the Hb subunits were able to form coordinate 1:1 complexes with heme, and these complexes bind oxygen in a manner expected for single subunit heme proteins. PMID- 15756817 TI - Immunogenicity of heme complexes of peptides designed to mimic the heme environment of myoglobin and hemoglobin. AB - In the preceding paper (Protein J. 25, pages 37-49, 2005), we reported the preparation and oxygen-binding properties of peptides that form stable complexes with heme mimic. The design of the peptides was based on the natural environment of the heme group in myoglobin (Mb) and in the alpha- and beta-subunits of human adult hemoglobin (Hb). In the present work, the heme-peptides were each administered into mice, either as emulsions in adjuvant (both for injections and boosters) or intravenously as solutions in phosphate-buffered saline. Antibody (Ab) responses, monitored up to 14 weeks after the first administration, showed that when the heme-peptides were injected with adjuvant they stimulated Ab responses against the immunizing peptide, which in most cases bound to the correlate protein (Mb or Hb). However these heme-peptides were non-immunogenic when administered in PBS intravenously. It is concluded that heme-peptides:(a) would not trigger an adverse immune response if used for transfusion purposes. PMID- 15756818 TI - Characterization of various recombinant antigens from Echinococcus multilocularis for use in the immunodiagnosis. AB - Using four clones isolated from Echinococcus multilocularis cDNA library with alveolar echinococcosis (AE) patient sera, various antigens were expressed as ThioHis tag-fused protein. Recombinant EmII/3 antigen was produced as the five fragments divided into the N-terminal (#5 and #5s), the central (#6 and #6s) and the C-terminal domain (#7). Immunoblot analysis revealed that the #7 showed significant reactivity whereas those of #5 and #5s were relatively low. The #6 and #6s also showed lower reactivity than that of #7, although the two minor bands of #6 reacted with every serum. These results suggested that an immunodominant region of EmII/3 locate within the C-terminal one third. The #8s recombinant antigen, Ser23-Glu176 of actin filament fragmenting protein (AFFP), apparently reacted with the AE patient sera, while the #1 antigen synthesized as a full-length antigen BI did not show such high reactivity. Thus, #7 and #8s antigens showed significant potential for use in immunodetection of AE. In addition, the specific antibodies against #7 and #8s reacted with specific antigens in crude extract of E. multilocularis cyst, indicating that these antigens retained antigenicity common to native EmII/3 and AFFP, respectively. PMID- 15756819 TI - New Fusarium wilts on vegetable crops in Italy. AB - Three Fusarium wilts recently observed in Italy on lettuce (Lactuca sativa), wild (Diplo taxis spp.) and cultivated (Eruca sativa) rocket and lamb's lettuce (Valerianella olitoria) emerged as major production problems in Lumbardy (north western Italy). Aspects of biology and epidemiology of the three diseases and some possibilities of disease management are discussed. PMID- 15756821 TI - Diversity of the Phytophthora infestans population in Flanders, Belgium. AB - A total of 94 isolates of Phytophthora infestans were collected from disease outbreaks in commercial potato crops and private gardens in 2002 and 2003. The isolates were recovered successfully from single lesions of diseased potato foliage. Not from all isolates pure cultures were obtained due to contaminations with Fusarium species and bacteria. The structure of the population was analysed phenotypically. Characteristics of the isolates included in vitro growth rate, mating type, in vitro sensitivity to the phenylamide fungicide metalaxyl-M and allozyme genotype at glucose-6-phosphate isomerase (Gpi) and peptidase (Pep) loci. Significant differences in in vitro growth rate were observed among the 52 isolates by comparing the main radial growth of the isolates after 7 days. Forty seven from the isolates tested were the Al mating type. Only one isolate was characterized as A2 mating type. Isolates with sensitive, intermediate and resistant responses to metalaxyl-M were detected in the populations. Forty isolates had a growth of less then 40 % at 5 ppm metalaxyl-M. Three isolates had a growth of less then 40 % at 100 ppm metalaxyl-M. Eight isolates had a growth of more then 40 % at 5 and 100 ppm metalaxyl-M. Cellulose acetate electrophoresis was used to examine Gpi and Pep banding pattern of the population of P. infestans attacking potato in Flanders. All the isolates tested produced the 100/100 Gpi isozyme electromorph. Five different allozyme genotypes of the Pep loci were identified: 92/92, 96/96, 100/100, 92/100, 83/100. PMID- 15756820 TI - Dead flower buds of pear: effect of tree growth control, and Alternaria alternata as causal agent. AB - Dead (dormant) flower buds of pear is an important phenomenon in pear production in the Netherlands. Vigourous or unbalanced tree growth and Pseudomonas syringae pv. syringae are mentioned as likely causes of dead flower buds. Several tree growth control treatments including ethephon. Regalis (Prohexadione-Ca) and root pruning were evaluated. Regalis increased disease incidence. The plant stimulant (foliar fertilizer) Resistim (potassium phosphonate) reduced disease incidence. Pseudomonas syringae pv. syringae was occasionally isolated from diseased flower buds. However, Alternaria alternata was nearly always isolated from diseased buds. Indicating a strong relation between this fungus and dead flower buds of pear. PMID- 15756822 TI - Development of molecular tests for the detection of ILAR and latent viruses in fruit trees. AB - The detection throughout the year of latent and ILAR viruses in fruit tress by classical serological tests appear to be unreliable. We have developed RT-PCR tests for a reliable detection of latent and ILAR viruses in fruit trees. These assays were then simplified to allow the direct use of crude plant extracts instead of total RNA preparations, and the analyses of pooled samples. In this way, such RT-PCR protocols are suitable for a routine diagnosis of latent and ILAR viruses in fruit tree certification. PMID- 15756823 TI - Olive verticillium wilt or dieback of olive in Iran. AB - During 2000--03, different areas in Zanjan, Golestan and Khorasan provinces were surveyed for the presence of olive dieback. Olive branches, leaves and roots showing typical symptoms and soil around the roots were collected for further study. Samples were surface-sterilized with sodium hypochlorite or ethanol and then cultured on PDA and Czapek media. Soil samples were diluted in ethanol-agar for fungal isolation and purification. Morphological characteristics of the fungal mycelium particularly phialide and spores identified the causal agent to be the soil-borne pathogen, Verticillium dahliae. The disease was present in all olive growing regions but it was severe in temperate and relatively humid regions such as Gorgan. Infection index of the disease varied between 5 to 30% with an average of 11.89+/-1.12 among various orchards in this area. The newly established orchards showed more infection than the older ones. A significant difference in disease incidence and severity were observed among olive cultivars of Michen, Roughani, Zard and Koronakei. The latter cultivar had the least amount of infection. Strains of V. dahliae isolated from olive trees had different morphological and pathogenicity characteristics. These strains had different growth rates in response to the optimum temperature of 20 or 25 degrees C. The number of fungal propagules per gram of air-dried soil ranged from 2 to 32 with an average number of 13.42+/-0.50. Regarding the number of propagules of V. dahliae in the soil and susceptibility of cultivars in the newly established orchards, it seems necessary to take serious control measures to prevent disease spread. PMID- 15756824 TI - Characterization of Pseudomonas pathovars isolated from rosaceous fruit trees in East Algeria. AB - A survey of bacterial diseases due to Pseudomonas on rosaceous fruit trees was conducted. In forty two orchards located in the Constantine region ( East Algeria). Pseudomonas isolates were identified on the bases of their cultural and biochemical characteristics . A total of fifty nine phytopathogenic bacteria were isolated from diseased pome and stone fruit trees. Thirty one strains comparable to Pseudomonas syringae pv. syringae were isolated from cherry (Prunus avium L.), plum (P. domestica L.), apricot (P. armeniaca L.), almond (P. dulcis L.) and pear trees (Pirus communis L.); sixteen strains comparable to Pseudomonas syringae pv. morsprunorum were obtained from samples of cherry and plum. Twelve strains of Pseudomonas viridiflava were isolated from cherry, apricot and peach (Prunus persica L.). PMID- 15756825 TI - Differences in susceptibility of winter wheat varieties for Fusarium species under Belgian growing conditions. AB - Fusarium head blight is an important disease of cereal crops caused by Fusarium species. It causes not only a reduction in yield, but most Fusarium species (F. graminearum. F. culmorum, F. avenaceum. F. poae) produce also a range of toxic metabolites such as deoxynivalenol (DON) and zearalenone (ZEA). The evaluation of Fusarium species was followed up under natural infection conditions during the growing seasons 2001--2002 and 2002--2003 in two varietal winter wheat experiments on the experimental farm of the Hogeschool Gent at Bottelare. Disease pressure, DON and ZEA content, different Fusarium species as well as growth and yield parameters were determined. In both years there were significant differences between the varieties concerning the susceptibility to Fusarium and the DON content. ZEA was not found in the kernels. The mean deoxynivalenol (DON) content was in 2002 (1,126 mg/kg) higher than in 2003 (0.879 mg/kg) although the mean disease severity was bigger in 2003 than in 2002 what means that the DON content was not always correlated with the disease severity. The Fusarium species most frequently identified in our two field trials (Bottelare) were F. graminearum and F. culmorum Varietal differences in susceptibility to Fusarium species and DON contamination could be detected. PMID- 15756826 TI - New, rare or remarkable microfungi in the Italian Alps (Carnic Alps)--part I- ascomycotina. AB - During our observations in the SE part of the Carnic Alps in the year 2003 we were able to collect and identify 35 ascomycetes on trees and dead wood. Among these one can find numerous ascomycetes of different orders e.g. Pyrenomycetes, Loculoascomycetes and Discomycetes. Some species like Botryosphaeria ribis GROSENLUCHER & DUGGAR on Ribes alpinum L., Dothiora pyrenophora (FR.) FR. on Sorbus aucuparia L., Gemmamyces piceae (BORTH.) CASAGO. on Picea excelsa (LAM.) LINK, Glomerella montana (SACC.) v. ARX & E. MULLER on Sesleria caerulea (L.) ARD, Hymenoscyphus immutabilis (Fuck.) Dennis on Alnus incana (L.) Moench, Hysterographium fraxini (PERS. Ex. FR.) de Not. on Fraxinus ornus L., Lachnellula willkommii (Hartig) DENNIS [= Trichascyphella willkommii (Hartig) NANNF.] on Larix decidua MILL.,Leptosphaeria lycopodina (Mont.) SACC. on Lycopodium annotinum L., Mollisia adenostylidis REHM. on Adenostyles glabra (MILL.) DC., Pezicula cinnamomea (DC.)SACC. [ana: Cryptosporiopsis quercina PETRAK] on Quercus robur L., Pyrenopeziza petiolaris (A. & S. Ex FR.) NANNF. on Acer pseudoplatanus L., Tapesia rosae (PERS.) FUCKEL on Rosa canina L., are new for this area. All specimen are deposited in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha. PMID- 15756827 TI - New, rare or remarkable microfungi in the Italian Alps (Carnic Alps)--part II- other microfungi. AB - In addition to the collection of Ascomycotina in the Carnic Alps (see New, rare or remarkable microfungi in the Italian Alps (Carnic Alps) part I ) we were able to treasure about 300 species of parasitic and saprophytic microfungi. Among them Basidiomycetes, Ascomycetes and Deuteromycets like Bostrichonema polygoni (UNGER) SCHROT. on Polygonum viviparum L., Chrysomyxa rhododendri DE BY on Picea abies (L.) KARSTEN, Coleosporium tussilaginis (PERS.) BERK. I=C. cacaliae OTTH.] on Adenostyles glabra (MILL.) DC., Dasyscyphus barbatus (KUNZE) MASSEE on Lonicera nigra L., Leptosphaeria coniothyrium (FUCKEL) SACC. on Rosa canina L., Leptotrochila brunellae (LIND) DENNIS on Prunella grandiflora (L.) SCHOLLER., Marssonina kriegeriana (BES.) MAGNUS on Salix reticulata L., Puccinia alpina FUCKEL on Viola biflora L., Puccinia maculosa (STRAUSS.) ROHLING and Erysiphe cichoracearum DC. On Prenanthes purpurea L., Septoria microsora SPENG. on Gentianella germanica (WILLD.) BORNER, Urocystis orobranches (FR.) FISCH. V. WALDH. on Orobranche gracilis SM., Urocystis violae (J. SOWERBY) A. FISCHER VON WALDHEIN on Viola biflora L. and Uromyces phyteumatum (DC.) UNG. on Phyteuma spicatum L. were dominant. All samples are located in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha. PMID- 15756828 TI - Mycodiversity on a dead stem of the giant hogweed--Heracleum mantegazzianum Sommer et Levier. AB - During our investigations on microfungi in the Ruhr area we were able to find the remarkable mycodiversity of different genera and species on dead stems of Heracleum mantegazzianum, a Neophyt for the European flora. Altemaria alternata (FR.FR.) KEISSL., Camarosporium spec., Cladosporium herbarum (PERS.) LINK, Colletotrichum dematium (PERS. Ex FR.) GOVE, Dactylella armandii YADV., Dasyscyphus mollissimus (LASCH.) DENNIS, Dendryphion comosum WALLR., Endophragmia hyalosperma (CORDA) MORGAN-JONES & COLE, Epicoccum purpurescens EHRENB., Gliomastix luzulae (FUCKEL) MASON, Hymenoscyphus herbarum (PERS.) DENNIS, Lasiosphaeria caudata (FUCKEL) MASON, Lophiostoma caulium (FR.) CES. & de NOT, Periconia byssoides PERS., Phoma complanata (TODE EX FR.) DESM., Phoma longissima (PERS.) WESLEND., Pirottaea cf. nigrostriata GRADDON, Pleospora herbarum (PERS.) RABENH. EX. CES. & de NOT, Pleurophragmium parvisporum (PREUSS) HOLUBOVA-JECHOVA, Pyrenopeziza chailletii FUCKEL, Rhizopus stolonifer (EHRENB. FR.) VUILL, Sclerotina sclerotiorum (LIB.) DE BY., Septofusidium herbarum (BROWN & SMITH) SAMSON, Torula herbarum (PERS.) LINK, Trichoderma koningii OUDEM, Volutella melaloma BERK. & BR. The sample is located in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha. PMID- 15756829 TI - New and remarkable microfungi in North Rhine Westphalia, Germany. AB - During our investigation on microfungi in North Rhine Westphalia in the years 2002 and 2003 we were able to collect and identify some new and rare species of microfungi as parasites and saprophytes on wild and ornamental plants. Some of these like Erysiphe elevata (BURILL.) U. BRAUN & S. TAKAMATSU COMB. NOV. [=Microsphaera elevata BURILL.] on Catalpa bignonioides WALT., Erysiphe syringae japonicae (U. BRAUN) U. BRAUN & S. TAKAMATSU [= Microsphaera syringae-japonicae U. BRAUN, M. aceris BUNKINA. KOMAROVSKIE CHTENIYA, Erysiphe acerina U. BRAUN & S. TAKAMATSU] on Acer campestre L. and Acer barinerve L., Mycosphaerella iridis (DESM.) SCHROET., Ectostroma iridis FR. and Volutella melaloma BERK. & BR on Iris pseudacorus L., Puccinia doronicella P. SYD. & SYD. on Doronicum columnae TEN., Ascochyta lamiorum SACC. S.L. I=A. phlomidis BUB. & WROB.) on Phlomis tuberosa L., Colletotrichum gloeosporides (PENZ.) SACC. on Passiflora coerulea L., Oidium hortensiae JOERST on Hydrangea macrophylla (THUNB.) SER., Puccinia horiana P. HENN. on Chrysanthemum vulgare (L.) BERNH., Lophodermium pinastri (SCHRAD.) CHEV., Leptostroma pinorum SACC., Sclerophoma pythiophila (CDA) HOHN., Lichenoconium boreale (KARST.) PETRAK. & SYD., Anthostomella formosa KIRSCHST. and Sphaeropsis sapinae (FR.) DYKO & SUTTON on Pinus nigra L. are new for Germany. All samples are located in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha. PMID- 15756830 TI - Reactions of some cucurbitaceous species Tozucchini yellow mosaic virus (ZYMV). AB - Zucchini yellow mosaic virus (ZYMV) is a widespread serious pathogen of cucurbitaceous plants. ZYMV was first detected in Hungary in 1995. Since then it has become one of the most dangerous viruses of the Cucurbitaceae family causing serious epidemics. The virus has many hosts, which - particularly perennial ones may play important role as virus reservoirs and infection sources in virus epidemiology. On the other hand wild weed species maybe sources of resistance to viruses. Our research was carried out on a total of 15 wild species from 8 genera (Cucumis, Cucurbita, Cyclanthera, Ecballium Momordica, Lagenaria, Zehneria, Bryonia). Test plants were mechanically inoculated with ZYMV. Local and systemic symptoms were determined and 5 weeks after inoculation DAS-ELISA tests were also carried out. Symptomless plants were reinoculated to Cucumis sativus cv. Accordia test plants. On the basis of the results we determined the percentages of infections and so we classified the test-plants into sensitive and resistance categories. On the basis of the results new host plants of ZYMV are the followings: Bryonia dioica, Cyclanthera pedata, Ecballium elaterium, Momordica balsamina, Momordica rostrata, and Zehneria scabra. Among them Momordica balsamina and Ecballium elaterium showed latent to ZYMV. Bryonia alba and Zehneria indica are especially remarkable, because they proved resistant to ZYMV on the basis of symptomatology and serology. Our results might have significant role in the field of research of host range, virus resistance and virus differentiation. PMID- 15756831 TI - Occurrence, distribution and relative incidence of mosaic viruses infecting field -grown squash in Tehran province, Iran. AB - Squash (Cucurbita pepo) belongs to Cucurbitaceae family. Every year Cucurbitaceae are planted world wide. They are one of the most important economic crops. Cucurbitaceae are threatened by viruses. Many viruses damage the plants of this family. Since nine viruses have been reported on squash from Iran. In this survey, during 2002--2003, to determine the distribution of Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV) and Watermelon mosaic virus (WMV), 466 samples were collected from squash field in Tehran province. Infected plants showing symptoms such as: mosaic, yellowing, deformation, shoestring of leaves and fruit deformation and yield reduction. Distribution of CMV, ZYMV and WMV were determined by DAS-ELISA. Thepercentage of ZYMV, WMV and CMV were 35.6, 26.1 and 25.1% respectively. Triple infection (CMV+ZYMV+WMV) were found in 6.4% of samples. ZYMV were found the most frequently the viruses. This is the first report of WMV on squash in Tehran province. PMID- 15756832 TI - Occurrence and distribution of lettuce mosaic disease in Tehran province from Iran. AB - Lettuce mosaic virus (LMV) Cucumber mosaic virus (CMV) and Tomato spotted wilt virus (TSWV) were identified in fields of Tehran province. In this study 452 leaf samples were collected from the fields throughout the Tehran province during 2002 and 2003. Distribution of Lettuce mosaic virus (LMV), Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV)and Arabis mosaic virus (ArMV) was determined with DAS-ELISA. Percentage of single Infection to LMV. CMV or TSWV was 20.58, 15.93 and 9.96% respectively. Also 15.28% of samples were co- infected with LMV+CMV, 8.19% with LMV+TSMV and 7.74% with CMV+TSWV. 4.65% of samples were Infected to all of these three viruses. LMV was found in 48.69%, CMV in 43.59% and TSWV in 30.54% of samples totally. Therefore LMV is major dominant agent of lettuce mosaic disease in Tehran province. This is the first report of occurrence of TSWV on lettuce in Iran and first report of CMV and LMV in Tehran province. PMID- 15756833 TI - Biological and molecular characterization of lettuce mosaic virus from Tehran province in Iran. AB - In this study, lettuce samples having LMV infection symptoms were collected from Tehran fields during 2003. Samples tested for LMV infection by immuno printing. Three positive samples in immuno printing collected and their characteristics were determined. In mechanical inoculation, these Isolates produced symptoms on Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Nicotiana benthamiana, Lactuca sativa cv. Mantilia and cv. Terocadero (which contains the mol1 resistance gene and susceptible respectively), but not cv. Salinas 88 (which contains the mol2 resistance gene). LMV was purified and LMV polyclonal antiserum was produced in rabbit by a series of intravenous and intramuscular injections, the precipitin titre of this antiserum was 1:1024. Gel double diffusion test (GDDT) was performed, and precipitin bands appeared. SDS-PAGE and western blotting showed the presence of coat protein 29 kDa. In IC-RT-PCR with on LMV specific primer pair, an approximately 1300 bp fragment was amplified. PMID- 15756834 TI - The occurrence of root rot and crown rot of rice in Gilan and Zanjan provinces, Iran. AB - Root rot and crown rot of rice is one of the important fungal diseases of rice in Gilan and Zanjan provinces, Iran. During 1999--2002, samples of plant and soil around the roots of infected rice plants were collected and used to identify the causal agent. Root and crown parts were surface sterilized with sodium hypochlorite and then cultured on PDA (potato dextrose agar), PPA (pepton pentacholoritobenzene agar) and CLA (carnation leaf agar) media. Soil samples prepared in water agar were used to isolate the pathogen. The causal agent was identified as Fusarium moniliforme. Colonies were initially white but turned violet to grey late. Microconidia were arranged in chain and macroconidia were cylindrical and long with 3-5 septa. The disease was severe in Zanjan province particularly along Ghezel Ozan river where the infection ranged from 70-80%. Root and crown rot was more prevalent in areas where Champa and Gerdeh were being cultivated continuously. On the other hand, Sadri cultivars had relatively less infection. Persistent cultivation of rice and seed sowing method intensified disease development and caused significant economic losses. PMID- 15756835 TI - Influence of inoculum density of Verticillium dahliae, temperature and relative humidity on epidemics of verticillium wilt of cotton in northern Iran. AB - During 1992--2003, frequency of Verticillium dahliae propagules, disease incidence and severity of verticillium wilt of cotton were determined in several cotton growing fields in Golestan province, northeastern Iran. Inoculum density varied among fields and different years ranging between 2-47 propagules/g of air dried soil with an average of 18.96+/-0.73. In addition, the pattern of diseased plants varied with type of field and year. Simple regression analysis showed a linear relationship between inoculum density of V. dahliae at planting time on one hand, disease incidence and severity for all years on the other. The straight line model described the increase in disease intensity index over the accumulated physiological time from sowing. The number of days above 28 degrees C (T) and the area under relative humidity (RH) had significant effects on inoculum density in soil (MS) and final disease development (Y) and fitted the Y = 65.840 - 0.0034 RH + 0.57 MS - 1.7T model with R2 = 0.859 and significant F-function (p<0.0001). PMID- 15756836 TI - Fertility status and distribution of mating type alleles of the rice blast fungus, Magnaporthe grisea in northern Iran. AB - This study was carried out using 155 monoconidial isolates collected from different areas of two major rice growing provinces in northern Iran, including 94 isolates from Guilan and 59 isolates from Mazandaran. Among 94 isolates from Guilan, 92 and two isolates recovered from rice and crabgrass (Digitaria sp.), respectively. All 61 rested isolates from Mazandaran were recovered from rice. All isolates were evaluated for in vitro sexual fertility and mating type status by pairing with Mat 1-1 and Mat 1-2 fertile standard hermaphrodite isolates including Br48 and Th12 (Mat 1-1) and KA9 and TH16 (Mat 1-2). Of 155 isolates, 98 (63.2%) were fertile and 57 (36.8%) were infertile and produced no perithecium when mated with standard isolates. Among 98 fertile isolates, 96 isolates were identified as Mat 1-1 and two isolates as Mat 1-2. All Mat 1-1 isolates were obtained from rice and two Mat 1-2 isolates obtained from crab grass. No Mat 1-2 isolate was identified from rice in this study. Both mating types were found in Guilan but all isolates recovered from Mazandaran were identified as Mat 1-1. Male fertility predominated in fertile Mat 1-1 and Mat 1-2 isolates from all sampling sites in northern Iran, and no female fertility was detected. This is the first report of existence of Mat 1-2 allele in Magnaporthe grisea population in Iran. PMID- 15756837 TI - Detection of fungal infectous agent of wheat grains in store-pits of Markazi province, Iran. AB - Wheat is an economic and important crop that provides approximately 20% of food calorie in the world. It is first crop in Iran and cultivated in the most areas of this country. Store-pit fungi make undesirable changes in quality and appearance of wheat grains. Even, some fungi produce different mycotoxins which are toxic to human and livestock's that use wheat grains as source of food. In this study, several samples were randomly collected from each of five store-pits located in different areas of Markazi Province including: Arak, Mahallat, Khomein, Saveh and Sarband. Grains were treated on PDA, and blotter, agar and washing test also used for isolating and detection of fungi. At least 100 grains per each sample were randomly used for each test and treatment. The fungi that determined in this study were Cochliobolus australiensis, Cladosporium herbarum, Epicoccum sp., Tilletia leavis, Aspergillus flavus, A. niger, A. fumigatus, Alternaria alternata, Alternaria sp., Penicillium italicum, P. digitatum, Fusarium sp., Rhizopus sp., Ustilago tritici, Scytalidium sp. Among these fungi the most isolates were belonged to Cladosporium, Alternaria, Rhizopus and Fusarium. Cladosporium herbarum was the most common in different sampling areas. Tilletia laevis and Ustilago tritici were just recovered in washing test. This study revealed that different fungi are associated with wheat grains in store pits in Markazi Province. Some of them like Aspergillus flavus normally produce aflatoxin, a very toxic and carcinogenic mycotoxin that is harmful for human. PMID- 15756838 TI - Changes in specific activities of peroxidase, chitinase and phenylalanine ammonia lyase and phenolic content in cucumber leaves inoculated with Podosphaera fusca, the causal agent of powdery mildew. AB - In this investigation, the effects of powdery mildew disease [caused by Podosphaera fusca (syn. Sphaerotheca fuliginea)] on specific activities of several defense-related enzymes and phenolic content were studied in cucumber leaves. Spore suspension of the fungus was sprayed on cucumber (cv. Super Dominus) plants in greenhouse and leaves from both inoculated and non-inoculated control plants were sampled at 0, 24, 48, 72 and 144 hours after inoculation (HAI). Spore germination and tissue colonization of P. fusca were microscopically studied on the inoculated surface of leaf samples. Further, Phenolic content (PHE) and specific activities of peroxidase (POX), chitinase (CHI) and phenylalanine ammonia-lyase (PAL) were spectrophotometrically measured in leaf extracts. Time-course of disease progress on the leaf surface showed that maximum spore germination occurred within 24 HAI and host penetration and disease development process began during 24-48 HAI. Evaluation of enzyme activities showed that POX specific activity in inoculated plants significantly increased at 72 HAI onwards and reached 2.5 times of that of control at 144 HAI. CHI specific activity showed a transient reduction in inoculated plants between 48-72 HAI and thereafter increased significantly in relation to control. PAL specific activity in inoculated plants was not significantly different from that of control. PHE in inoculated plants showed a significant increase compared to control at 48 HAI and thereafter. Comparison of time-course of disease progress with changes in enzyme activities indicated that POX activity had an increasing trend during disease progress whereas CHI activity showed a transient decrease at the early stages and then increased during the later stages of infection: PAL activity did not show any changes during the infection and PHE increased at the early stages of infection process and remained constant at rest of the time. PMID- 15756839 TI - Effects of Acibenzolar-S-methyl on the specific activities of peroxidase, chitinase and phenylalanine ammonia-lyase and phenolic content of host leaves in cucumber-powdery mildew interaction. AB - One of the properties of systemic acquired resistance in plants is its concomitance with the biochemical changes including enhancement of activities of defense-related enzymes. In this study, the effects of Acibenzolar-S-methyl (ACI) on the some of defense responses of the cucumber plants (healthy or inoculated with spore suspension of Podosphaerafusca, the causal agent of cucumber powdery mildew) were surveyed via in vivo tests. Changes of defense responses in ACI treated cucumber plants, inoculated with pathogen or not, were studied and compared with those of non-treated control plants. Results indicated that specific activity of peroxidase increased significantly in treated plants. Increase in enzyme activity was higher in pathogen-inoculated than non-inoculated plants, thus pathogen attack stress to plant plays a role in enhancement of enzyme activity. Specific activity of phenylalanine ammonialyase showed no changes in ACI-treated non-inoculated plants, but in inoculated plants it Increased due to interaction between ACI treatment and pathogen attack stress. Specific activity of chitinase increased in both inoculated and non-inoculated ACI-treated plants at 24 hours after treatment onwards, and pathogen attack stress did not affect it. Phenolic content of ACI-treated plant tissues, despite of small fluctuations, did not show any definite pattern of changes. PMID- 15756840 TI - Model for total evaluation of a phytosanitary intervention in the context of new methodology of <>. AB - The model of the "GREAT FILIERA" is proposed for as it concerns the total evaluation of a phytosanitary intervention in the vine-growing and wine-producing sector. This model is essentially based on the application of a new methodology used for the evaluation of a control system (Carbonneau & Cargnello, 2003) already tested in applicatory phase. The total evaluation is determined by an appropriate panel of judges. They have considered and assigned a "weigh" in % in one scale of priority (number that follows the variable) and beyond fifty descriptors of the productive system, constituting the total system. PMID- 15756841 TI - Total, technical, economic and social evaluation of a plan for defense of the vineyard according to the methodology of the "GREAT CHAIN". AB - In another work, proposed in this congress, we have presented the basic methodology of the "GREAT CHAIN" in which a plan of fight to vine diseases is evaluated in the globality using 54 variables, to which a weight (in %) in a scale of priority at technical, economic, social and ethical level has been given. This basic methodology has been favourably accepted at the XIII GESCO Congress (Groupe International Systemes De Conduite de la Vigne)--February 2003 in Montevideo (Uruguay), and at the congress on "Paysages de Vignes et de Vins" in Fonteyraud (Val de Loire) (Carbonneau, Cargnello, 2003). In this work the results of researches on the total, technical, economic, social and ethical evaluation of a conventional phytopathologic defence plan led in a vineyard of the Latium according to the method of the "GREAT CHAIN" are exposed. These researches have shown that the model of conventional phytopathologic struggle, applied in the Latium, against pests of vine positively answers for how much concerns the plants protection: the answer is good at enterprise level, but negative for all that is defence and safeguard for man and environment, in the widest sense of the term. The researches have gone on the phytopathologic defence of biological and eco-compatible vineyards with very encouraging results. PMID- 15756842 TI - Severe outbreaks of tomato yellow leaf curl Sardinia virus in Calabria, Southern Italy. AB - During the winter 2003--2004 a serious disease was observed in protected tomato crops in Castrovillari, Reggio Calabria province, Southern Italy. Symptoms consisted in marginal leaf yellowing, leaf curling, plant stunting, flower abortion. The disease was detected in a group of greenhouses (about 10ha) where several tomato cultivars were grown hydroponically. The highest incidence of infection (60-100%) was observed in tomatoes grafted on Beaufort DRS tomato rootstock. Since the symptoms were similar to those described for Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV), detection assays for these viruses were used. In DAS-ELISA positive results were obtained with a abroad-spectrums reagent combination (distributed by Bioreba AG) detecting TYLCV, TYLCSV, and other begomoviruses. When DNA probes were used in tissue print assays, positive reactions were obtained for TYLCSV, but not for TYLCV. The two probes consisted of digoxigenin-labelled DNAs representing the coat protein gene of either TYLCSV or TYLCV. Attempts to isolate the viral agent by mechanical inoculation failed, except in few cases where Potato virus Y and Tobacco mosaic virus were identified following transmission from symptomatic plants to herbaceous indicatorpplants. By contrast, grafting onto tomato seedlings always successfully transmitted the disease. In the Castrovillari area TYLCSV was not reported before. The rootstocks that nurseries used for grafting were obtained from Sicily, where the disease is endemic and both TYLCSV and TYLCV are widespread. Probably the grafted plantlets represented the primary source of infection from which subsequent diffusion by way of the vector Bemisia tabaci followed. In fact the vector had previously been detected in both the glasshouse grown and open field tomato crops in Calabria region. TYLCV was previously reported in a different area of Calabria in 1991, but apparently it was an occasional outbreak, and B. tabaci was not detected. Since in the Castrovillari area surveyed in the present study tomato is grown throughtout the year in protected crops, the whitefly vector of the virus is present, and some natural hosts of the virus are found, it is feared that TYLCSV may become endemic, as already happened in Sicily, Sardinia, and Spain several years ago. In Spain and Sicily TYLCV, together with TYLCSV, was reported as the causal agent of very severe tomato crop losses. Therefore the danger exists that also TYLCV will reach this area, furthermore complicating the management of tomato crops. PMID- 15756843 TI - Evaluation of oat cultivars and lines under infection with barley yellow dwarf virus. AB - Thirteen domestic and foreign oat cultivars and eight breeding lines bred from the University of Illinois were evaluated for resistance to barley yellow dwarf (BYD) using artificial inoculation with Rhopalosiphum padi viruliferous for an isolate of Barley yellow dwarf virus-PAV endemic to Moscow region origin. Cultivar Blaze and six Illinois lines showed the best grain yields under disease pressure that resembled a BYD epidemic. PMID- 15756844 TI - Resistance to air-born diseases and pests induced by compost in substrate cultivation of strawberry. PMID- 15756845 TI - In situ development and application of cDNA-AFLP to isolate genes of Candida oleophila (strain O) potentially involved in antagonistic properties against Botrytis cinerea. AB - The yeast Candida oleophila (strain O) presents a high level of protective activity against Botrytis cinerea (gray mold) on postharvest apples. The cDNA AFLP technique allows the comparison of mRNA populations extracted from cells grown in different conditions. In order to isolate yeast genes potentially involved in biological control properties, that technique was applied on strain O cells growing on apple wounds. The biological control properties of 8 C. oleophila strains and strain O were assessed in order to compare the gene expression of a non antagonistic strain against gene expression of strain O. In the absence of a non-antagonistic strain, an other comparison model was designed. It was based on the growth of strain O in different in situ conditions: strain O applied on apple wounds (O), strain O applied on apple wounds in presence of B. cinerea (B) and B. cinerea alone on apple wounds (F). A recovering technique, based on the washing of cells in the wound and a RNA extraction method followed by a DNase treatment were optimised before cDNA-AFLP application. Thirteen primer pairs were used. Their application resulted in an average of 54 and 55 bands for O and B respectively whereas no bands were observed for F. Among these bands, 8 were expressed more intensely in presence of the pathogen (1.1% of the fragments). PMID- 15756846 TI - Efficacy assessment of Candida oleophila (strain O) and Pichia anomala (strain K) against major postharvest diseases of citrus fruits in Morocco. AB - Two yeasts, Candida oleophila (strain O) and Pichia anomala (strain K), were previously selected for their antagonistic activity against postharvest diseases on apples and pears. The objective of the study was to determine the efficacy of both antagonistic yeast's against wound postharvest pathogens of citrus fruits. The efficacy of both strains (applied at 10(5), 10(6) and 10(8) CFU/ml) was assessed against Penicillium digitatum and P. italicum inoculated after one hour (at a concentration of 10(5), 10(6) and 10(7) spores/ml) on citrus varieties 'clementine' and 'valencia-late'. Fruits were incubated for one week at 24 degrees C before measurement of lesion diameter. The protective levels were positively correlated with high concentration of antagonist and low concentration of pathogen. Highest protective levels (from 73 to 100%) were detected with the application of strain O or strain K at 10(8) CFU/ml whatever the pathogen (applied at 10(5) spores/ml) and the citrus variety. The antagonistic activity of both strains was also dependent on the incubation period before pathogen Inoculation. The protective level increased with time between application of the antagonist and inoculation of fungal spores. Whatever the yeast strain (10(8) CFU/ml). the protective level exceed 70% when wounded oranges were inoculated with P. digitatum or P. italicum (both at 10(6) spores/ml) 12 hours after yeast treatment. These protective levels reached 100% when the incubation period separating the antagonist application and the pathogenic inoculation was 24 hours. On the other hand, high protective levels (< 80%) were also observed against the sour rot decay on citrus variety 'clementine' caused by Geotrichum candidum inoculated at concentration of 10(6) spores/ml when strain O or strain K were applied at 10(8) CFU/ml 24 hours before pathogen. All these results support the potential practical application of both strains against major postharvest pathogens on citrus. PMID- 15756847 TI - Results of screening greenhouse and int vitro behaviour to evaluate some durum wiieat varieties and their F5 descendents selected in algeria for resistance to Septoria tritici. AB - 15 genotypes of durum wheat, composed of 5 parental varities and 10 F5 lines selected in Algeria, were studied for evaluating thein behaviour to Septoria tritici Two trials were realised: one under green house conditions, where seedlings were inoculated artificially with Septoria tritici suspension, and another under in vitro conditions where detached leaves where also inoculated. Among this range of varieties, some genotypes were tolerant, others susceptible and others showed rather reactions between tolerant and susceptible. Tolerant genotypes presented a low number of necrosis in the green house trial and a reduced lesion size on the in vitro infected tissu. On the contrary. sucseptible genotypes showed a high necrosis number and an important lesion size on their detached leaves. Some observations, during the experimentation, permitted to underline the important limiting effect that could impose some biotic and abiotic factors (temperature, relative humidity and age of plant tissue) to the pathogen development, reported by other studies before, In comparision with other investigations, varieties which were reported to be resistant or susceptible had confirmed their reactions in our study, it will be interesting to include them into a primary range of varieties for use to study the fungus Septoria tritici. PMID- 15756848 TI - Study of the effect of Ammoides pusilla (Brot.) Breist, essential oil against Pseudomonas sp. AB - The economic losses caused by plant pathogens, the development of genetic resistance to applied pesticides by major pathogenic bacteria and public concerns about the use of synthetic chemicals on food and environment, increased interest in the search of alternative safe methods to control diseases spreading. Biological control with plant extracts has emerged as a promising option. The present study was carried out to determine the effect of Ammoides pusilla essential oil on the growth of two pathogenic bacteria, Pseudomonas syringae pv. syringae and Pseudomonas syringae pv. mosprunorum Three essential oil treatments, 1/2 v/v, 1/5 v/v and 1/10 v/v concentrations, were tested onto these bacteria, using the agar diffusion test. The results showed that 1/2 v/v concentration treatment completely inhibited Pseudomonas syringae pv. mosprunorum growth and produced a 35 mm diameter inhibition zone for Pseudomonas syringae pu. syringae. 1/5 v/v essential oil concentration produced inhibition zones of 19 and 17 mm diameter for Pseudomonas syringae pv. syringae and Pseudormonas syringae pv. mosprunorum respetively, while 1/10 v/v essential oil concentration gave inhibition plates of 11 and 12 mm diameter for both stains in the order cited. In in vitro culture essential oil gave promising results suggesting that future studies should be carried out and focus on determining appropriate formulation of this bio-molecule to be use as a bio-control agent. PMID- 15756849 TI - The effect of solanapyrone a produced by Ascochyta rabiei on seed germination and the elongation of radicles and hypocotyls of chickpea (Cicer areitinum L.). AB - Three Algerian isolates of A. rabiei (72, Mat 1.2 and 9216) were grown on Czapek Dox medium supplemented with cations and incubated for 14 days. After incubation, the mycelium of the fungus was removed by filtration through four layers of muslin cloth and spores were removed from the filtrate by centrifugation at 10,000 g for 20 min. Solanapyrone A was partially purified by liquid phase extraction into ethyl acetate and, after removal of the ethyl acetate, the toxin samples were dissolved in methanol and quantified by analytical High Performance Liquid Chromatography (HPLC). Solanapyrone A was identified by superimposition of its UV spectrum, obtained from the diode array detector of the HPLC, on the spectrum of an authentic sample. The action of solanapyrone A solution on seed germination and elongation of radicles and hypocotyls was tested using a concentration of 18.2 microg/ml and a two-fold dilution series of this solution in distilled water. The three Isolates, 72, Mat1.2 and 9216 produced solanapyrone A at concentrations of 37.2, 14.2 and 11.09 microg/ml, respectively. When probit % inhibition of seed germination was plotted against log2 of solanapyrone A concentration, there was a linear relationship and the EC50 concentration was determined as 7.2 microg/ml. Similarly, when radicle and hypocotyl elongation was plotted against log2 of solanapyrone A concentration, both gave linear relationships and the EC50 concentrations were determined as 5.37 and 6.02 microg/ml, respectively. It was concluded that solanapyrone A has a considerable inhibition of chickpea. However radicles and hypocotyls were susceptible than seed germination. PMID- 15756850 TI - Biocontrol agents of Botrytis cinerea tested in climate chambers by making artificial infection on tomato leafs. AB - To reduce the use of chemical agents, that are causing damage to the environment, in the fight against Botrytis cinerea, different BCA's were tested for their possibility to control Botrytis cinerea in a biological way. In order to investigate the effectiveness of the different micro organisms and Elot-Vis, experiments were carried out in climate chambers with 5 weeks old tomato plants. Leafs on the plant were inoculated with drops of a suspension that contained spores of Botrytis cinerea. The possible antagonists that were tested in these experiments were Trichoderma harzianum (Trichodex), T. asperellum (Biofungus), T. hamatum (T382), Bacillus subtilis (Serenade and Phytovit) and Pseudomonas aeruginosa (7NSK2 and KMPCH). For all these different micro organisms the direct and the indirect influence on Botrytis cinerea was investigated. In tests where the direct influence of the antagonists was examined, the spores of the moulds or the bacteria were suspended together with spores of Botrytis cinerea and subsequently drops of this suspension were pippeted on the leafs. After a while by ideal circumstances for Botrytis cinerea the infections on the inoculated leafs were counted. For the indirect influence of the antagonists, also leafs of 5 weeks old tomato plants were inoculated with a suspension of Botrytis cinerea spores. The roots of the tomato plants that were used for testing the indirect influence were treated during there growth with a suspension of the antagonist to see if induced systemic resistance pathway (ISR) was activated. For testing the effectiveness of Elot-Vis, tomato plants were sprayed a few times with a solution of this product during their growth. Results of the climate chamber test of the plants that were treated with Elot-Vis, showed a reduction of Botrytis cinerea infections on the inoculated leafs. Biological control agents seem to be not always very effective against Botrytis cinerea. The biological control agents that are containing micro organisms are also depending on the circumstances of the environment for an optimal development. These conditions are not always that optimal to compete with Botrytis cinerea or other micro organisms that are present on the plant. PMID- 15756851 TI - Different products for biological control of Botrytis cinerea examined on wounded stem tissue of tomato plants. AB - For the moment the agents that are used against Botrytis cinerea, in glasshouses were tomatoes are cultivated, are from chemical origin. For reducing the use of chemical agents in the future it is important to search for effective biological control agents against the fight of Botrytis cinerae. The following biological products Vital pasta, Vital gel and Elot-Vis were examined in there possibility to control Botrytis cinerea. Elot Vis was tested out in experiments that were carried out in climate chambers were leafs of 3 week old tomato plants were artificially infected with Botrytis cinerea spores. Also the biological products of Vital were first investigated in experiments that were carried out in climate chambers. In stead off leafs of tomato plants it were stem wounds of tomato plants who were treated with the pasta or the gel that was spread over the wounded surface after this has been inoculated with a suspension of conidia of Botrytis cinerae. The results of these first tests that were executed in the climate chambers were the circumstances for Botrytis cinerea were ideal seemed promising. In the next step these products were tested out on large scale in glasshouses. For each plant 5 wounds were created by removing the leafs, these wounds were or first treated with the Biological product and thereafter artificially infected with Botrytis cinerea spores to check out if these products can be used as a preventive agent or the wounds were first inoculated with a suspension of Botrytis cinerea spores and thereafter treated with the product. For the product Elot-Vis a few plants were totally sprayed with an Elot-Vis suspension before leafs were removed and the wounds were inoculated with conidia of Botrytis cinerea to check out if this product was able to activated the induced systemic resistance pathway. The experiments that were executed in glasshouses showed different percentages of succeeded Botrytis cinerea infections. This is probable due to the different weather conditions during both days that the experiments were executed. For the wounds that were treated with pasta it was difficult to distinguish wounds were Botrytis cinerea succeeded to infect the plant, because these wounds frequently didn't show any sign of infection on the surface but when the wounds with the pasta were cut open it was possible to see Botrytis cinerea infections inside the stem. PMID- 15756852 TI - Biological control of chickpea Fusarium wilt by antagonistic bacteria under greenhouse condition. AB - In this survey, Fusarium oxysporum was isolated from roots infected plants and was shown to be pathogenic. Experiment were carried out with seven antagonistic bacteria. Based on biochemical, Physiological and morphological tests, isolates B 120, B-32, B-28 and B-22 were identified as Bacillus subtilis and isolates Pf 100, Pf-10 and CHAO as Pseudomonas fluorescens. In greenhouse studies, only isolate B-120 (Less than benomyl) reduced Fusarium wilt of chickpea in both seed and soil treatments. The application of antagonistic bacteria had no different effects on plant growth factors. Soil treatment of bacteria had a better effects on plant growth than that of bacterial seed treatment. The use of antagonists (B 120, B-28, B-120 and CHAO) in combination had no significant effect on plant growth factors and reduction wilt disease than that each isolate was applied individually. PMID- 15756853 TI - Effects of inoculum density and substrate type on saprophytic survival of Phytophthora drechsleri, the causal agent of gummosis (crown and root rot) on pistachio in Rafsanjan, Iran. AB - In this study, sampling was carrid out on soils around pistachio trees in various regions of Rafsanjan, Iran. Following isolation and identification of Phytophthora isolates, the predominant species was found to be P. drechsleri and used for further investigation. For studying saprophytic survival of the fungus, soils collected from different areas were combined and autoclaved. Sterile soil was divided into 10 parts and mixed with fungal inoculum at various concentrations of 0-9% (w/w) separately. Each soil part (100g) was placed in 15cm diameter plastic pot. Some soils in pots were supplemented with sterile wheat straw whereas others were mixed with pistachio leaves surface sterilized with 5% (v/v) sodium hypochlorite. After 3, 6 and 9 weeks of incubation, five leaves or straws samples were taken from each replicate and cultured on CMA-PARPH medium and the fungal colony formation was monitored. The experiment was performed using completely randomized design with factorial experiments including three factors (substrate type, inoculum density and time), 10 treatments (0-9 g inoculum levels) and nine replicates. The results showed that the type of substrate (wheat straw and pistachio leaf) was very important for the fungal saprophytic survival in that this was significantly greater for the pistachio leaves. Time was also considered another critical factor for the fungal survival. With passing incubation time, saprophytic survival of the fungus declined. Further, it was demonstrated that increasing inoculum density would result in longer survivability of P. drechsleri and maximum fungal survival on substrate was obtained when inoculum density was at 9% (w/w). PMID- 15756854 TI - Antagonistic effects of several bacteria on Fusarium oxysporum, the causal agent of root and crown rot of onion under field conditions. AB - Onion (Allium cepa) is one of the most important vegetable crop which is commonly used as a food supplement. This plant is found to be vulnerable to various pathogenic infections during its growth development. Among different onion diseases, root and crown rot,caused by Fusarium oxysporum f.sp. cepa, s considered an importantfungal disease. In this study, the inhibitory effect of Bacillus cereus (isolates 22 and 52), B. subtilis (isolate 126), Pseudomonas fluorescens (isolates 48 and CHAO), benomyl fungicide and a combination of isolates CHAO and 22 and isolate 52 and benomyl were investigated on disease development under the field condition. This experiment was carried out in a randomize complete blocks with 10 treatments and three repetitions. Grouping of treatments was done at 5% level using Duncan multiple comparison test. It was also demonstrated that isolate 126 was the most effective antagonist with regard to crop yield but other treatments despite showing significant on plant growth factors were less effective in increasing crop yield. PMID- 15756855 TI - Evaluation of chickpea resistance against some isolates of Ascochyta rabiei, the causal agent of ascochyta blight. AB - Ascochyta blight, incited by Ascochyta rabiei (Pass.) Lab., is one of the most important fungal diseases that has been shown to cause significant yield losses on chickpeas in Kermanshah province, Iran. One of the most effective and reliable methods of controlling the disease has been the use of resistant cultivars worldwide. This requires the identification of the physiological races of the fungus in the region in advance. In this regard, pathogenicity of 30 isolates of A. rabiei was determined using 10 differential chickpea varieties (mean of 1-9 degree of disease in three replications) under greenhouse conditions. In this respect, isolates fell into four groups, whereas based on infection type three groups were identified using cluster analysis. Disease resistance of four chickpea cultivars, Hashem, Jam, ILC-482 and Bivanij, were evaluated against isolates from each of three groups separately and in combination in the greenhouse and field conditions (with 4 replications). The experiment was repeated twice with three replications in greenhouse. Chickpea cultivars were sprayed with fungal spore suspension (2x10(5) spores/ml). Results showed that Jam and Bivanij cultivars were susceptible under greenhouse as well as field conditions. Hashem was tolerant to group I and susceptible to groups II and III and in the field. ILC-482 was tolerant to groups II and I and susceptible to group III and the combination of groups under greenhouse condition. PMID- 15756856 TI - The assessment of the rice cultivars/lines resistance to blast disease in Mazandaran province, Iran. AB - Blast, caused by Magnaporthe grisea, is one of the most important diseases in rice production regions of the world including Iran. To determine progress of rice blast disease on the selective cultivars and lines also to assay some components of partial resistance, a set of Iranian rice cultivars (Local and breeding) along with near-isogenic lines (NILs) and breeding lines from International Rice Research Institute (IRRI) were tested with some field races of the fungus in blast nursery and five of selective races in greenhouse. These experiments were conducted in a Randomized complete Block Design (RCBD) with three replications (except greenhouse experiment on the leaves). Traits in this study consisted of Infection Neck Number (INN), Neck Lesion Size (NLS), Infection Type (IT), percent Diseased Leaf Area (DLA) and Area Under Disease Progress Curve (AUDPC); also IT, Sporulation Lesion Number (SLN), Sporulating Region Diameter (SRD) and percent DLA were measured in leaf blast in greenhouse (one replication). The Iranian local cultivars and NILs i.e. Co-39 and C104-PKT located as susceptible group for AUDPC, IT, INN and NLS. Iranian breeding cultivars, breeding lines from IRRI and NILs (except Co-39 and C104-PKT) were resistant or indicated hypersensivity reaction (HR). Some cultivars (Fujiminori, Onda, and Hassan Saraii) were semi susceptible to leaf blast in nursery. The main point is correlation in 1% (a = 0.0001) between the traits in greenhouse and blast nursery. Neck node infection of Haraz cultivar in greenhouse experiment to IA-89 race is very important, because Haraz is a resistant cultivar to blast disease in Iran. PMID- 15756857 TI - Activity of natural products against courgette powdery mildew. AB - The effectiveness of several natural products in field trials were tested against powdery mildew on courgette. Sulphur, sodium bicarbonate (alone or in mixture with pinolene, mineral oil or sulphur), sodium silicate in mixture with mineral oil, potassium permanganate, lecithin, Equisetum arvense in mixture with Timus vulgaris, Ampelomyces quisqualis and Reynoutria sachalinensis were evaluated. Generally, in our working conditions (very strong pathogen infection), only some products were effective. Good results were obtained using sulphur and potassium permanganate in the first year and very interesting results about disease control were obtained, in the second year of the trial, using sulphur alone or in mixture with sodium bicarbonate. The products tested did not show any phytotoxic symptoms. PMID- 15756858 TI - Repercussion of the phytosanitary treatments on various models of foliage management in a Cabernet Sauvignon Vineyard (I note). AB - This study is the first communication about the repercussion of several phytosanitary treatments in three different training systems, differently managed in foilage, in relation to the usual phytopatologies: botrytis, acid rot, grape mildew and oldium. Particular attention to residuals of the products supplied on the foilage and present in ground and wine is paid. The considered foilage management systems are among those more spread and effective ones for a good quality wine production for Cabernet Sauvignon variety (clone R5). 11. Cordon Spur of Conegliano (C.S.C); 12. Simple Free Espalier Curtain (S.C.S.L); 13. Lyra (Lyra). The phytosanitary interventions for the foilage management systems have been compared between the company's ordinary plan and one with various and numerous active principles. The CG-ECD with Multiresidual analysis has determined the active principles residuals in the ground and in the wines. The characteristics of the wines have been determined by physical- chemical analysis; their organoleptic quality has been valued by panels of producers, oenologists, consumers and restaurant staff. The results point out a difference among the trials for the different presence of residuals. The foilage management justifies the diversity of the analytical composition of the wines. There is a substantial uniformity of judgements for the organoleptic quality. In conclusion, in this prove the judgement on the economic quality highlights the uselessness of the use of more incisive and radical active principles. PMID- 15756859 TI - Biological control of Phytophthora ramorumon rhododendron. AB - Phytophthora ramorum was found in Poland in 2000 as the causal agent of rhododendron blight. Besides eradication of diseased plants and rhododendron growing around, chemical and biological control of the pathogen is necessary. In this study in vitro activity of grapefruit extract and chitosan in the inhibition of P. ramorum growth and sporulation and their efficacy in the control of leaf and stem rot development was evaluated. Amendment of V8 juice agar and soil leachate with grapefruit extract resulted in the inhibition of colony growth and sporulation of P. ramorum. Zoosporangia were more susceptible to the extract than pathogen hyphae and chlamydospores. Chitosan only slightly inhibited the colony growth and zoosporngia production. Spraying of rhododendron inoculated with P. ramorum with grapefruit extract at conc. 165 microg/cm3 inhibited 2-3 times the spread of necrosis on stems and leaves. Pre- and postinoculation spraying of rhododendrons with chitosan at conc. of 1000 microg/cm3 suppressed the disease spread about 40%. PMID- 15756860 TI - Effectiveness of some means using against root rot on parsley seedling root. AB - The results of a two-year study concern the effectiveness of 4 substances- biopreparate Chitosol (beta-1,4 D-glucosamine polymer) and fungicides: Rovral Flo 255 SC (BAS iprodione 255 g/dm3), Sportak Alpha 380 EC (BAS prochloraz 0,300 dm3/dm3 and carbendazim 80 g/dm3) and Zaprawa Funaben T (carbendazim 20% + tiuram 45%)--used against root rot and plant rot was described. These experiments were conducted in glasshouse and field conditions. Root of parsley cultivar Berlinska were treated one of tested substances autumn before storage period in clamp or spring immediately before planting seedling roots. The results showed that autumn dressing was ineffective, indices of root infestation were similar in each combination including control in both years of observation. Spring dressing was better, but not only in controls were many parsley plants decaying in the fields. Among substances used for spring dressing of seedling roots, the best efficacy exhibited Zaprawa Funaben T and Sportak Alpha 380 EC. PMID- 15756861 TI - Oils activity against Melampsora epitea on willow. AB - Willow trees cv. Iwa growing in open field conditions in the sandy soil and naturally infected with Melampsora epitea were used. Effectiveness of paraffin oil (Atpolan 80 EC) and plant oils (Olejan 85 EC, rape and sunflower) in the control of pathogen was evaluated. When first uredinia were seen on the lower leaf surface, plants were sprayed 2-times at 7-day intervals with tested compounds at concentration from 0.25 to 2%. Also Surfactant Tergitol (TM) 15-S-9 at conc. 0.3% was added to plant oil suspension. After 2 sprayings a number of uredinia per leaf and number of browned and destroyed uredinia were counted. In the first trial Atpolan 80 EC suppressed uredinia formation about 2-4-times whereas rape or sunflower oil about 1.8 to 3-times. Additionally about 11 to 30% of uredinia were dried. In the next trials effectiveness of tested oils was higher than in previous experiment. After 2-weeks of willow trees protection with Atpolan 80 EC formation of uredinia was inhibited from 7 to 17-times, whereas Olejan 85 EC suppressed their production about 4 to 14-times. Additionally about 9-62% (Atpolan 80 EC) or 10-25% (Olejan 85 EC) of uredinia were browned and dried depend on used concentration. In the third experiment after 2-week-protection of plants with Atpolan 80 EC inhibition of uredinia formation varied from 6 to 10 times. In the case of Olejan 85 EC about 2 to 10-times less uredinia were observed on protected plants than on control, untreated leaf blades. About 48-72% (Atpolan 80 EC) or 17-61% (Olejan 85 EC) of uredinia were dried. PMID- 15756862 TI - Chitosan (biochikol 020 PC) in the control of some ornamental foliage diseases. AB - Chitosan, (Biochikol 020 PC) a potential elicitor of plant defence and also an active inhibitor of fungal growth was used in experiments. The compound was used at concentrations 0.01 to 0.2% as a plant spray for rose protection against Sphaerotheca pannosa var. rosae, Peronospora sparsa and Diplocarpon rosae, Oidium chrysanthemi and Puccinia horiana on chrysanthemum, against Melampsora epitea on willow. Myrothecium roridum on dieffenbachia and against Lophodermium spp on Scots pine. Effectiveness of this product, applied curatively 2-times at 7-day intervals after appearance of rose powdery mildew symptoms on most of plant parts, was about 32%. When compound was applied just after the first disease symptoms appearance, after 2-week-protection its effectiveness was about 43-60%. After 4 weeks, effectiveness of chitosan (Biochikol 020 PC) increased and ranged from 67 to 77%. In the control of P. sparsa on rose shrubs, growing in plastic tunnel, chitosan at conc. 0.025% was applied 4 times at weekly intervals. Effectiveness of chitosan against this pathogen was over 72%, similar as standard fungicide. Increase of chitosan (Biochikol 020 PC) concentration resulted in decrease of its effectiveness. In case of D. rosae control, chitosan used 9 times at weekly intervals was applied after first disease symptoms. After 3-week protection effectiveness of the compound varied from 18 to 60% according to used concentration. After 9 weeks of protection effectiveness of tested product ranged from 16 to 23% and was connected with used concentration. Increase of chitosan concentration resulted in better protection of rose shrubs. Effectiveness of chitosan (Biochikol 020 PC) used 4-times at conc. 0.01 to 0.05% as chrysanthemum spray in the control of Oidium chrysanthemi ranged from 69 to 79%, whereas against Puccinia horiana from 54-97%. Two-time-spraying of willow with rust symptoms (Melampsora epitea) with chitosan at conc. 01-0.04% caused decrease of disease severity from 26 to 47%. Additionally about 10 to 25% of uredinia were dried up. Chitosan (Biochikol 020 PC) applied for leaf spraying inhibited the development of Myrothecium leaf spot on dieffenbachia more than 85%. Curative application of the compound controlled Myrotecium leaf spot at about 40%. Chitosan efficacy was also tested against one-year-old Pinus sylvestris seedlings growing in field conditions. Plants were sprayed fortnightly 14-times (long programme) or 5-times (short programme) against Lophodermium spp. Observations made during next spring showed that effectiveness of chitosan (Biochikol 020 PC) was more than 50% (long programme) and below 50% (short programme). PMID- 15756863 TI - Limitation of point source pesticide pollution: results of bioremediation system. AB - Groundwater and surface water is at risk of contamination from the use of some agricultural pesticides. In many circumstances pesticide contamination of water resources is more likely to result from point sources than from diffuse sources following approved application to crops in the field. Such point sources include areas on farms where pesticides are handled, filled into sprayers or where sprayers are washed down. To overcome this way of contamination different kind of bio-remediation systems are nowadays in development. In Flanders, Belgium two pilot plants of bioremediation systems for the in situ retention and/or degradation of pesticides were installed. Both systems were based on the Phytobac concept, a watertight excavation filled with straw, peat, compost and soil. The channel was made in the bottom from plastic foil. All kinds of spray rests were captured by the phytobacs. This study focuses on what level pesticides leach, bio degrade or are retained by the filling of the phytobac. The soil-properties of the filling were investigated. Pesticide tracers were added for monitoring to both phytobacs. Soil and water samples were taken during one year. Pesticides are retained at least for one month by the filling of the phytobac. Almost no pesticide leached out. In winter hardly any pesticide degradation was observed in the filling of the phytobac. In summer no detectable pesticides were still left in the phytobacs. PMID- 15756864 TI - Pesticides re-entry dermal exposure of workers in greenhouses. AB - This research has the aim to evaluate the risk of pesticide dermal exposure for workers in greenhouses. We considered the following crops: tomato, cucumber and strawberry, largely spread in Bracciano lake district. The pesticides monitored were: tetradifon on strawberry: metalaxyl, azoxystrobin and fenarimol on cucumber; acrinathrin, azoxystrobin and chlorpyrifos ethyl on tomato. The dermal exposure was evaluated by Dislodgeable Foliar Residue (DFR) measurements employing transfer coefficients got from literature. For risk evaluation, we have compared the dermal exposures with Acceptable Operator Exposure Levels (AOEL). The re-entry time were obtained intercepting the dose decay curves with AOEL values. The re-entry times result higher than two days in the cases of chlorpyrifos on tomato (re-entry time: 3 days), azoxystrobin on tomato (4 days), and tetradifon on strawberry (8 days). The need of measuring specific transfer coefficients is pointed out. PMID- 15756865 TI - Signum, a new fungicide with interesting properties in resistance management of fungal diseases in strawberries. AB - Signum, a new fungicide developed by BASF, was applied during 6 successive years against fungal diseases in strawberries. The product is formulated as a water dispersible granule, containing 6.7 % pyraclostrobin and 26.7 % boscalid. Pyraclostrobin is similar in chemistry to other strobilurin fungicides like kresoxim-methyl and trifloxystrobin, registered for fruit disease control. Boscalid belongs to the class of carboxyanilides. Both components in the premix formulation combine two different biochemical modes of action in the fungal cell respiration. Therefore, this co-formulation gives a broad-spectrum activity and also a reduced resistance risk for different target pathogens. Botrytis cinerea is the most important disease on strawberry-fruits and thus the control of fruit rot is mainly focused on this fungus. In average over 6 years, Signum has not only given a very good control against Botrytis fruit rot, but it has also shown a high performance in the control of Colletotrichum. Besides, Signum provides good control of powdery mildew (Podosphaera aphanis) and limits the shift to other fruit rots like leather rot (Phytophthora cactorum and leak (Rhizopus, Mucor). The availability of several categories of fungicide families with a different mode of action gives opportunities in alternating different fungicides and is the best guarantee for a sustainable control of fruit rot in all kinds of strawberry production methods. Signum should be integrated in an overall disease management program. Trials, in which the applications of Signum were timed on disease forecasting, based on environmental factors favorable for Botrytis development, were very promising. This tool can also help in establishing the IPM concept in the production of strawberries. PMID- 15756866 TI - Effect of different chemical and mechanical defolation methods on the skin quality of potatoes. AB - Without foliage destruction an efficient harvest is impossible. Potatoes for the fresh market are often harvested when the foliage is still heavy green due to tuber size and starch content that must be limited. Tubers from immature vines are typically very susceptible to skinning and mechanical injury during harvest. Young tubers from immature vines need more time after foliage destruction to set periderm than tubers from senescent vines where the formation of periderm is already started. Spray schemes based on metoxuron, carfentrazone-ethyl and diquat at a dose of 300 g/ha caused slower leaf and stem desiccation. Over the 3 growing seasons it could be concluded that mechanical foliage destruction in combination with carfentrazone-ethyl + mineral oil promoted periderm formation better than the other desiccation schemes tested. A split treatment with diquat at 300 g/ha or carfentrazone-ehtyl + mineral oil followed by a second application of diquat or carfentrazone-ethyl can led to a slower periderm formation and even give secondary growth. A double treatment of diquat (300 g/ha) or carfentrazone-ethyl + mineral oil followed by diquat (600 g/ha) after 3 days gave satisfactory results. Rhizoctonia tuber infection increased with a longer field period after treatment. In general the increase was more pronounced for the spray schemes where skin set of the tubers was less fast. PMID- 15756867 TI - Pesticides in the Cagayan valley (Philippines): usage, drift patterns and the exposure of farmers differing in income and market access. AB - Over the past ten years, the amount and number of different types of pesticides have increased significantly, which led to a growing concern about the possible adverse effects on human health and the environment. This is particularly true for countries where regulations are not strictly implemented and farmers' knowledge of safe handling is often inadequate. This paper discusses the results of a series of spray experiments to determine drift patterns along field boundaries and the exposure of farmers during their usual spraying exercises. Moreover, farmers' pesticide usage and methods of application will be described, and the effects of income and market accessibility on pesticide use patterns will be investigated. It is based on a study conducted in four villages located at increasing distance from the national highway leading to regional markets and connecting the Cagayan Valley in Northeast Luzon with Manila. The 20 pesticides encountered in this study cover 18 different active ingredients, 9 of which are classified by the WHO as 'highly hazardous' or 'moderately hazardous'. The EPA has classified at least 6 of the encountered pesticide formulations as Restricted Use Pesticides. Nevertheless, all pesticides are freely sold in stores or on markets and applied by farmers without personal protection in an unsafe manner. The farmers living nearest to the highway have the highest income and largest farms. Yet they are most at risk, having easiest access to pesticides and spraying the largest quantities of pesticides per hectare, compared to the farmers living at greater distance from the highway. It is recommended to review the list of pesticides approved for use in the Philippines and discern between Restricted and General Use Pesticides. Several recommendations for improving the implementation of pesticide policies and the IPM program are given. PMID- 15756868 TI - Identification of two fungicide degrading Pseudomonas species by gas chromatography of cellular fatty acids. AB - Microorganisms can be identified in many ways. Conventional methods rely on the expression of certain properties that are usually mediated directly by enzyme activity. Extension of this approach to include numerical identification or automated systems to analyze results often strengthens conclusions. Identification of bacterial species using morphological characters and biochemical tests is often difficult and time consuming. Immunodiagnostic and nucleotide hybridization techniques have improved sensitivity, specify, precision, and ease of testing. Chemotaxonomy is also precise and can result in the definition of highly discriminatory properties. Cellular Fatty Acids (CFA) analyses fall into this category. One of the most convenient methods for the identification of fatty acids in bacterial cells is by gas-liquid chromatography of their methyl esters prepared from phospholipids, total lipids, or other lipid fractions. Two bacterial strains from Bahar Yossof. Al-Fayiurn governorates, Egypt tentatively identified as a species of pseudomonas by virtue of its physiological and biochemical characteristics. Confirmation of this identification was carried out using fatty acids profile analysis. PMID- 15756869 TI - Fungicidal activity of new N-alkyl and N-aryl Chitosan derivatives. PMID- 15756870 TI - Insecticidal and fungicidal activity of new N,O-acyl Chitosan derivatives. PMID- 15756871 TI - Residues of profenofos in spring onion. AB - Profenofos is one of the commonly used insecticides in the control of Trips tabaci on spring onion in Iran. Residues of profenofos in spring onion were determined in two different fields under the same conditions. In the first field, onion plants were sprayed with profenofos (40EC) at the rate of 1000 g/ha. Spraying was repeated 2 weeks later. In the second field one spraying was preformed at the same rate. Spring onionwere sampled at different time intervals and analyzed for profenofos residues using a GC equipped with NPD detector. In the first field's samples, the residues were 0.097 and 0.025 mg/kg at 2 and 6 days after spraying, respectively. The residues declined to 0.002 mg/kg on the day twelve. Two days after the second spraying the residues was 0.27 mg/kg, which reduced to 0.032 on the day sixth. However the residues were not detectable 32 days after the second spraying. In the second field, residue levels were 0.193 and 0.043 mg/kg at 2 and 6 days after spraying. Residues, which found after 32 days, were less than 0.001 mg/kg. The rate of residue decay in the first field was higher than the second field. PMID- 15756872 TI - Teratogenicity study of some pesticides in chicken embryos. AB - The use of pesticides involves the risk of poisoning on wild animals. Teratological tests carried out on avian embryos provide useful data for environmental protection and facilitate the development of environment-friendly chemical plant protection techniques. A 30% dimethoate containing insecticide formulation (BI 58 EC) and a 20% benfluralin containing herbicide formulation (Flubalex) and a 960 g/l S-metolachlor containing herbicide formulation (Dual Gold 960 EC) were studied in chicken embryos after single administration by immersion and injection technique. Treatment was done on day 0 of incubation. Applied concentration of pesticides were 0.1% (dimethoate) and 2.05% (S metolachlor) and 0.375% (benfluralin) corresponding to that used in plant protection practice. Test materials were injected into the air chamber in a volume of 0.1 ml/egg, or eggs were treated by the immersion technique for 30 min. at 37 degrees C. Evaluation was done on day 19 of incubation. Injection treatment: the administration of S-metolachlor and benfluralin did not result a significant decrease in the average body weight of embryos. At the same time the body weight of embryos significantly decreased because of single administration of dimethoate. The embryomortality increased markedly after the administration of test materials (S-metolachlor, benfluralin, dimethoate). Immersion treatment: the administration of S-metolachlor and benfluralin and dimethoate did not result a significant decrease in the average body weight of embryos. The rate of embryomortality was low after the administration of S-metolachlor, benfluralin and dimethoate. After the immersion and the injection treatment the incidences of developmental anomalies were sporadic. In summary it can be established that the injection treatment was more toxic than immersion technique of the test materials in our study. PMID- 15756873 TI - Irritative effects of some pesticides and a technical component on tissue structure of the chorioallantoic membrane. AB - The chorioallantoic membrane (CAM) is a complete tissue that responds to injury with a complete inflammatory reaction, this process similar to that induced by chemicals in the conjunctival tissue of the rabbit eye. During the study chemicals are placed directly onto the chorioallantoic membrane and the occurrence of vascular injury or coagulation in response to a compound is as an indication of the potential of a chemical to damage mucous membranes. In our study irritant pesticides (Fusilade S, Karathane LC) and a technical pesticide component (Trend) were tested and their effects on the tissue structures of CAM were examined. After treatment with the test materials, first lysis and then haemorrhage were observed macroscopically on CAM. In histological pictures stained with H-E the rupture of the blood vessel wall was seen and blood was observed around the blood vessels in the middle layer. The histological findings correlated well with the macroscopic appearance in this study. In general a good correlation was found between the HET-CAM results and reported data from Draize test. The subjective nature of the evaluation is reduced through the histological examination of treated CAM. The HET-CAM test can be a useful component of a battery of tests needed for replacing the Draize rabbit eye irritation test. PMID- 15756874 TI - Toxicity of a dichlorvos containing insecticide formulation and an atrazine containing herbicide formulation in chicken embryos after individual administration. AB - A 50% dichlorvos containing insecticide formulation (Unifosz 50 EC) and a 50% atrazine containing herbicide formulation (Hungazin PK 50 WP) were studied in chicken embryos after administration as single compounds. Applied concentrations of dichlorvos were 0.1% (corresponding to the plant protection practice), 0.05%, 0.02%, 0.01%. Applied concentrations of atrazine were 0.66% (corresponding to the plant protection practice), 0.33%, 0.132%, 0.066%. The test materials were injected directly into the air-chamber of eggs on day 0 of the hatching period and evaulation was carried out on day 19 of incubation. The chicken embryos were examined for the following: rate of embryo mortality, body mass, type of developmental anomalies. After the single administrations of dichlorvos containing insecticide formulation and atrazine containing herbicide formulation on day 0 of incubation, the average body weight of chicken embryos significantly did not decrease as compared to the control. After the individual administrations of pesticides the incidence of developmental anomalies was sporadic. The embryonic mortality markedly increased at the highest concentrations of pesticides. The rate of embrio mortality were 61% (dichlorvos insecticide containing formulation) and 52% (atrazine containing herbicide formulation). In summary, the 50% dichlorvos containing insecide formulation (Unifosz 50 EC) and the 50% atrazine containing herbicide formulation (Hungazin PK 50 WP) were toxic to the developing chicken embryos at the highest concentration in our study. The toxic effect was expressed in the high rate of embrio mortality. PMID- 15756875 TI - The correlation of the spraying volume with herbicide adherence and herbicide penetration in glyphosate treatments. AB - The relationship between the spraying volume and glyphosate effectiveness has been widely studied over the years. Likewise, the effect of different adjuvants on glyphosate leaf adherence/penetration is currently well understood. However, not much is known about the way spraying volumes may influence these two herbicide properties or, in other words, which factor (adherence or penetration) is the limiting factor when spraying at low or high volumes of treatment. Using grass-weed Lolium rigidum and broad-leaved weed Portulaca oleracea as experimental systems, the effect of low (200 L ha(-1)) and high (800 L ha(-1)) spraying volumes and six commercial adjuvants (poly-l-p-menthene, a mixture of methyl oleate and palmitate, alkylglycol ester, dodecylbenzene ammonium sulphonate, and two paraffinic oils) plus ammonium sulfate on the effectiveness of five commercial glyphosate formulations has been studied under laboratory controlled conditions. Dose-response assays showed significant differences between low and high spraying volume treatments as expected, the former being much more effective than the latter. However, and in all commercial formulations and weed species tested, the best results at low spraying volumes were always obtained when formulating glyphosate with oils or their derivatives, with a minimal improvement observed in glyphosate/surfactant mixtures. On the contrary, both the use of ionic or non-ionic surfactants as glyphosate adjuvants gave the best results at high spraying volumes, with only a marginal success observed on paraffinic oils and fatty acidesters. Data point out glyphosate absorption as being a two-step process made up of adherence first and then penetration, both events being essential for glyphosate effectiveness. Under these terms, high performance glyphosate-rich low spraying volumes succeed by themselves in soaking leaf surfaces, penetration therefore being the main and only limiting factor, while low performance glyphosate-poor high spraying volumes fail to either soak or penetrate leaf cuticles. The use of penetrative agents such as oils is therefore advised as a single method to increase glyphosate effectiveness at low spraying volumes, whereas glyphosate treatments at high spraying volumes are better improved by using surfactant agents or surfactant + oil mixtures. PMID- 15756876 TI - Biochemical indicators of hepatotoxic effects of pesticides. AB - Pesticides can cause damage to man and beneficial organisms. Some sub-lethal effects of pesticides were studied in birds with a view to identifying characteristic biochemical responses that may be useful for the monitoring of exposure to sub-lethal levels in the field. Pesticides were used: demeton-S methyl, (DSM), chlorpyriphos, chlorfenviphos, triazophos, pirimicarb, methiocarb and permethrin. Blood was collected before dosing, and 2, 6, 24, 48 and 72 hours after the treatment from the brachial vein of birds. Enzyme activities were assayed in the plasma or serum samples obtained. The assays used were GOT, MDH, GDH, SDH, GAMMA GT and ChE. The results showed an increase in plasma and serum GOT and gamma-GT levels were found in all animals treated with the previous pesticides. The level of ChE increased in birds after treatment with permethrin. It was concluded that the pesticides cause structural and functional changes in the liver and also, the measurement of the previous parameter activities may be useful for assessing exposure and sub-lethal effects of pesticides on the wildlife. PMID- 15756877 TI - Wind tunnel evaluation of several tracer and collection techniques for the measurement of spray drift. AB - In the history of pesticide drift measuring techniques, different tracers and a lot of different collection techniques have been used. At the start of a new Flemish project 'Protecting the Flemish environment against drift - The importance of drift-reducing techniques', wind tunnel tests have been executed to select the most efficient tracer and collection technique. As tracer types a fluorescent tracer Renaissance W15, 2 different chelates, a NaCl-solution and a fungicide Tolylfluanide were used. 2 different collection techniques were tested: drains incorporated in the wind tunnel floor filled with filter paper and filled with cloths. The recovery of the different tracers combined with the 2 collection materials was calculated. The advantages and disadvantages of the tracers and collection materials are enumerated in this article. PMID- 15756878 TI - Risk of transmission of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) from patients colonized with MRSA. PMID- 15756879 TI - Society for Healthcare Epidemiology of America guideline approach works to control a methicillin-resistant Staphylococcus aureus outbreak. PMID- 15756880 TI - High prevalence of carriage of methicillin-resistant Staphylococcus aureus at hospital admission in elderly patients: implications for infection control strategies. AB - BACKGROUND: Despite contact isolation precautions for patients with methicillin resistant Staphylococcus aureus (MRSA), MRSA infections are increasing in many countries. OBJECTIVE: To evaluate the role of a potential unrecognized reservoir of MRSA carried by patients in acute care wards, we determined the prevalence of MRSA at hospital admission, with special emphasis on screening-specimen yields. SETTING: A 1100-bed teaching hospital in Paris, France. METHODS: Nasal screening cultures were performed at admission to a tertiary-care teaching hospital for patients older than 75 years. RESULTS: MRSA was isolated from 63 (7.9%) of 797 patients. On the multivariate analysis, variables significantly associated with MRSA carriage were presence of chronic skin lesions (adjusted odds ratio [AOR], 5.10; 95% confidence interval [CI95], 2.52-10.33); transfer from a nursing home, rehabilitation unit, or long-term-care unit (AOR, 4.52; CI95, 2.23-9.18); and poor chronic health status (AOR, 1.80; CI95, 1.02-3.18). Without admission screening, 84.1% of MRSA carriers would have been missed at hospital admission and 76.2% during their hospital stay. Furthermore, 81.1% of days at risk for MRSA dissemination would have been spent without contact isolation precautions had admission screening not been performed. CONCLUSIONS: MRSA carriage at hospital admission is far more prevalent than MRSA-positive clinical specimens. This may contribute to failure of contact isolation programs. Screening cultures at admission help to identify the reservoir of unknown MRSA patients. PMID- 15756881 TI - Trends in methicillin-resistant Staphylococcus aureus (MRSA) bloodstream infections: effect of the MRSA "search and isolate" strategy in a hospital in Italy with hyperendemic MRSA. AB - OBJECTIVE: To evaluate the secular trends in MRSA BSIs after the introduction of a nosocomial MRSA control intervention. DESIGN: Before-after study. SETTING: An 850-bed community hospital with an ICU and vascular surgery, neurosurgery, bone marrow transplantation, and AIDS units. MRSA is endemic at this hospital; the prevalence of methicillin resistance among patients with S. aureus infection is greater than 50%. PATIENTS: Among all inpatients, MRSA BSI was identified, its origin defined, and incidence rates calculated by ward and origin. INTERVENTION: A MRSA control program was implemented based on active surveillance cultures to identify MRSA-colonized patients, followed by isolation using contact precautions. Incidence rates of MRSA BSI during the intervention (i.e., July 1, 1997, to December 31, 2001) and preintervention (i.e., January 1, 1996, to June 30, 1997) periods were compared. RESULTS: Sixty-nine MRSA BSIs were identified. When compared with the preintervention period, the incidence rate of MRSA BSI was reduced from 0.64 to 0.30 per 1000 admissions (RR, 0.46; CI95, 0.25-0.87; P = .02) during the intervention period. The impact was greater in the ICU, with an 89% reduction (RR, 0.11; CI95, 0.01-0.98; P = .03), and for CVC-associated MRSA BSIs, with an 82% decrease (RR, 0.17; CI95, 0.05-0.55; P = .002). Methicillin resistance among S. aureus blood isolates decreased from 46% to 17% (RR, 0.36; CI95, 0.22-0.62; P = .0002). CONCLUSION: A reduction in MRSA bacteremia is achievable through use of the MRSA "search and isolate" intervention even in a hospital with high rates of endemic MRSA. PMID- 15756882 TI - Nosocomial acquisition of methicillin-resistant Staphylococcus aureus during an outbreak of severe acute respiratory syndrome. AB - OBJECTIVE: The four hospitals assessed in this study use active surveillance cultures for methicillin-resistant Staphylococcus aureus (MRSA) and contact precautions for MRSA-positive patients as part of routine infection control practices. The objective of this study was to determine whether nosocomial acquisition of MRSA decreased in these hospitals during an outbreak of severe acute respiratory syndrome (SARS) when barrier precautions were routinely used for all patients. DESIGN: Retrospective cohort study. SETTING: Three tertiary care hospitals (a 1100-bed hospital; a 500-bed hospital; and an 823-bed hospital) and a 430-bed community hospital, each located in Toronto, Ontario, Canada. PATIENTS: All admitted patients were included. RESULTS: The nosocomial rate of MRSA in all four hospitals combined during the SARS outbreak (3.7 per 10,000 patient-days) was not significantly different from that before (4.7 per 10,000 patient-days) or after (3.4 per 10,000 patient-days) the outbreak (P = .30 and P = .76, respectively). The nosocomial rate of MRSA after the outbreak was significantly lower than that before the outbreak (P = .003). Inappropriate reuse of gloves and gowns and failure to wash hands between patients on non-SARS wards were observed during the outbreak. Increased attention was paid to infection control education following the outbreak. CONCLUSIONS: Inappropriate reuse of gloves and gowns and failure to wash hands between patients may have contributed to transmission of MRSA during the SARS outbreak. Attention should be paid to training healthcare workers regarding the appropriate use of precautions as a means to protect themselves and patients. PMID- 15756884 TI - Spread of a methicillin-resistant Staphylococcus aureus ST80-IV clone in a Danish community. AB - OBJECTIVE: We report a community cluster of methicillin-resistant Staphylococcus aureus (MRSA) in Denmark with emphasis on routes of transmission and infection control measures. The objective is to extend knowledge of MRSA in a community setting where a nosocomial link could effectively be ruled out. DESIGN: Population-based observational study from November 1997 until June 2003. SETTING: North Jutland County, with approximately 495,000 inhabitants. SUBJECTS: The cluster encompassed 46 individuals and 26 households. INTERVENTIONS: Infection control measures included repeated visits to affected households by an infection control nurse who undertook screening for carriage among all household members and provided a program for decolonization. RESULTS: The causal strain was identical to a newly described international clone, ST80; SSCmec type IV; and Panton-Valentine leukocidin positive. Plausible routes of transmission included household contact and contact at work, kindergarten, and school. We did not detect a nosocomial source or any secondary cases in hospitals. Transmission by healthcare contact outside the hospital was plausible for three cases. We found evidence that the clone was introduced on more than one occasion to immigrant families from the Middle East. A 5-day decolonization program was successful at first attempt in 15 of 16 households that could be evaluated. CONCLUSIONS: Despite the described infection control measures, we continued to see new cases, underlining a need for a national policy to contain MRSA in the community. PMID- 15756883 TI - Management of inpatients colonized or infected with antimicrobial-resistant bacteria in hospitals in the United States. AB - BACKGROUND: Although guidelines for multidrug-resistant organisms generally include recommendations for contact precautions and surveillance cultures, it is not known how frequently U.S. hospitals implement these measures on a routine basis and whether infectious diseases consultants endorse their use. METHODS: The Emerging Infections Network surveyed its members, infectious diseases consultants, to assess their use of and support for contact precautions and surveillance cultures for routine management of multidrug-resistant organisms in their principal inpatient workplace. Specifically, members were asked about use of these strategies for methicillin-resistant Staphylococcus aureus, vancomycin resistant enterococci, and multidrug-resistant, gram-negative bacilli on general wards, ICUs, and transplant units. RESULTS: Overall, 400 (86%) of 463 respondents supported the routine use of contact precautions to control one or more multidrug resistant organisms in at least one unit, and 89% worked in hospitals that use them. In contrast, 50% of respondents favored routine use of surveillance cultures to manage at least one multidrug-resistant organism in any unit, and 30% of respondents worked in hospitals that use them routinely in any unit. Members favored routine use of surveillance cultures significantly more in ICUs and transplant units than in general wards for each multidrug-resistant organism (P < .001). CONCLUSIONS: Most of the infectious diseases consultants endorsed the use of contact precautions for routine management of patients colonized or infected with multidrug-resistant organisms and work in hospitals that have implemented them. In contrast, infectious diseases consultants are divided about the role of routine surveillance cultures in multidrug-resistant organism management, and few work in hospitals that use them. PMID- 15756885 TI - Epidemiology and clonality of methicillin-resistant and methicillin-susceptible Staphylococcus aureus causing bacteremia in a tertiary-care hospital in Spain. AB - OBJECTIVES: To describe the relative proportions of nosocomial and community onset Staphylococcus aureus bacteremia at our institution and the epidemiologic characteristics and clonal diversity of S. aureus isolates, as determined by pulsed-field gel electrophoresis (PFGE) and antimicrobial resistance patterns. DESIGN: Retrospective cohort study of all cases of S. aureus bacteremia between October 2001 and October 2002. SETTING: A 1300-bed, tertiary-care hospital. RESULTS: One hundred sixty-two unique episodes of S. aureus bacteremia were identified. Forty-three cases (26.5%) were caused by methicillin-resistant S. aureus (MRSA). Most cases of S. aureus bacteremia, whether MRSA or methicillin susceptible (MSSA), were nosocomial in origin (77.2%) or were otherwise associated with the healthcare system (16%). Only 11 (6.8%) of the cases (all MSSA) were strictly community acquired. Thirty-five unique macrorestriction patterns were identified among the 154 isolates that were typed by PFGE. Four major genotypes were defined among the isolates of MRSA, with 36 (85.7%) represented by a single PFGE type. Of the isolates within this major clone, all (100%) were ciprofloxacin resistant and 77.8% were erythromycin resistant. In contrast, the 112 isolates of MSSA comprised 31 different PFGE types, 3 of which represented 42.9% of all MSSA isolates and were associated with both nosocomial and community-onset bacteremia. CONCLUSIONS: Most cases of S. aureus bacteremia in our healthcare region are nosocomial in origin or are acquired through contact with the healthcare system and are thus potentially preventable. To preclude dissemination of pathogenic clones, it is therefore necessary to redouble preventive measures in both the hospital and the community. PMID- 15756886 TI - Molecular surveillance of clinical methicillin-resistant Staphylococcus aureus isolates in neonatal intensive care units. AB - BACKGROUND AND OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) has become an important nosocomial pathogen in our neonatal intensive care units (NICUs) and accounts for almost all S. aureus clinical isolates. The objective of this study was to assess the relatedness of these MRSA strains. DESIGN: MRSA clinical isolates were collected from infants hospitalized in our NICUs. Pulsed field gel electrophoresis with SmaI digestion was used to fingerprint these isolates. SETTING: Level-III NICUs in a university-affiliated children's hospital. RESULTS: Between 1998 and 2000, a total of 122 MRSA clinical isolates were collected from 104 infants hospitalized in our NICUs. Fifteen infants had multiple isolates (range, 2 to 4 isolates). The sources of specimens included blood (72), pus (23), sputum (15), body fluids (3), and catheter tips (9). A total of 4 genotypes with 20 subtypes were identified. There were 2 genotypes in 1998, 2 genotypes in 1999, and 4 genotypes in 2000. All but 2 isolates belonged to either genotype A (63.1%; 7 subtypes) or genotype C (35.2%; 11 subtypes). Among the 15 infants with multiple isolates, the genotypes of the isolates from a single episode of MRSA infection were different in 2 of 12 cases, and reinfection with a new strain was noted in 3 of 5 cases with recurrent infections. CONCLUSIONS: Two predominant MRSA clones prevailed in our NICUs between 1998 and 2000. Polyclonal bacteremia and reinfection with a new strain were noted PMID- 15756887 TI - Detection and treatment of antibiotic-resistant bacterial carriage in a surgical intensive care unit: a 6-year prospective survey. AB - OBJECTIVE: To describe, during a 6-year period, multidrug-resistant bacterial carriage in an intensive care unit (ICU). DESIGN: Prospective survey of 2235 ICU patients with methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E). SETTING: A surgical ICU in a tertiary-care teaching hospital. PATIENTS: All admitted patients. INTERVENTIONS: Nasal and rectal swabs were performed at admission and weekly thereafter. There was nasal application of mupirocin for MRSA carriers and selective digestive decontamination with local antibiotics for ESBL-E carriers. RESULTS: The swab compliance rate was 82% at admission and 51% during ICU stay. The rates of MRSA carriage or infection were 4.2 new cases per 100 admissions and 7.9 cases per 1000 patient-days during ICU stay. The rates of ESBL-E carriage or infection were 0.4 new case per 100 admissions and 3.9 cases per 1000 patient days during ICU stay. Importation of MRSA increased significantly over time from 3.2 new cases per 100 admissions during the first 3 years to 5.5 during the last 3 years. The rate of ICU-acquired ESBLE decreased from 5.5 cases per 1000 patient days during the first 3 years to 1.9 cases during the last 3 years. Nasal and digestive decontamination had low efficacy in eradicating carriage. CONCLUSIONS: MRSA remained poorly controlled throughout the hospital and was not just a problem in the ICU. MRSA thus requires more effective measures throughout the hospital. ESBL-E was mainly an ICU pathogen and our approach resulted in a clear decrease in the rate of acquisition in the ICU over time. PMID- 15756888 TI - The impact of methicillin resistance in Staphylococcus aureus bacteremia on patient outcomes: mortality, length of stay, and hospital charges. AB - OBJECTIVE: To evaluate the impact of methicillin resistance in Staphylococcus aureus on mortality, length of hospitalization, and hospital charges. DESIGN: A cohort study of patients admitted to the hospital between July 1, 1997, and June 1, 2000, who had clinically significant S. aureus bloodstream infections. SETTING: A 630-bed, urban, tertiary-care teaching hospital in Boston, Massachusetts. PATIENTS: Three hundred forty-eight patients with S. aureus bacteremia were studied; 96 patients had methicillin-resistant S. aureus (MRSA). Patients with methicillin-susceptible S. aureus (MSSA) and MRSA were similar regarding gender, percentage of nosocomial acquisition, length of hospitalization, ICU admission, and surgery before S. aureus bacteremia. They differed regarding age, comorbidities, and illness severity score. RESULTS: Similar numbers of MRSA and MSSA patients died (22.9% vs 19.8%; P = .53). Both the median length of hospitalization after S. aureus bacteremia for patients who survived and the median hospital charges after S. aureus bacteremia were significantly increased in MRSA patients (7 vs 9 days, P = .045; 19,212 dollars vs 26,424 dollars, P = .008). After multivariable analysis, compared with MSSA bacteremia, MRSA bacteremia remained associated with increased length of hospitalization (1.29 fold; P = .016) and hospital charges (1.36 fold; P = .017). MRSA bacteremia had a median attributable length of stay of 2 days and a median attributable hospital charge of 6916 dollars. CONCLUSION: Methicillin resistance in S. aureus bacteremia is associated with significant increases in length of hospitalization and hospital charges. PMID- 15756889 TI - Costs and outcomes among hemodialysis-dependent patients with methicillin resistant or methicillin-susceptible Staphylococcus aureus bacteremia. AB - OBJECTIVE: Comorbid conditions have complicated previous analyses of the consequences of methicillin resistance for costs and outcomes of Staphylococcus aureus bacteremia. We compared costs and outcomes of methicillin resistance in patients with S. aureus bacteremia and a single chronic condition. DESIGN, SETTING, AND PATIENTS: We conducted a prospective cohort study of hemodialysis dependent patients with end-stage renal disease and S. aureus bacteremia hospitalized between July 1996 and August 2001. We used propensity scores to reduce bias when comparing patients with methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) S. aureus bacteremia. Outcome measures were resource use, direct medical costs, and clinical outcomes at 12 weeks after initial hospitalization. RESULTS: Fifty-four patients (37.8%) had MRSA and 89 patients (62.2%) had MSSA. Compared with patients with MSSA bacteremia, patients with MRSA bacteremia were more likely to have acquired the infection while hospitalized for another condition (27.8% vs 12.4%; P = .02). To attribute all inpatient costs to S. aureus bacteremia, we limited the analysis to 105 patients admitted for suspected S. aureus bacteremia from a community setting. Adjusted costs were higher for MRSA bacteremia for the initial hospitalization (21,251 dollars vs 13,978 dollars; P = .012) and after 12 weeks (25,518 dollars vs 17,354 dollars; P = .015). At 12 weeks, patients with MRSA bacteremia were more likely to die (adjusted odds ratio, 5.4; 95% confidence interval, 1.5 to 18.7) than were patients with MSSA bacteremia. CONCLUSIONS: Community-dwelling, hemodialysis dependent patients hospitalized with MRSA bacteremia face a higher mortality risk, longer hospital stays, and higher inpatient costs than do patients with MSSA bacteremia. PMID- 15756890 TI - Prevalence and nosocomial spread of methicillin-resistant Staphylococcus aureus in a long-term-care facility in Slovenia. AB - OBJECTIVES: To determine the prevalence and incidence of methicillin-resistant Staphylococcus aureus (MRSA) colonization among residents and healthcare workers (HCWs) of a long-term-care facility (LTCF), to assess possible routes of nosocomial spread, and to determine genetic relatedness of the isolates. SETTING: A 351-bed community LTCF for the elderly. DESIGN AND PARTICIPANTS: Study investigators made two visits, approximately 3 months apart, to the facility. Samples for cultures were obtained from 107 residents during the first visit, 91 residents during the second visit, and 38 HCWs. RESULTS: The prevalence of MRSA colonization among residents was 9.3% during the first visit and 8.8% during the second visit. During the first visit, two HCWs were colonized. During the second visit, no HCWs were colonized. The colonization of HCWs suggested a potential role in the transmission of MRSA. Molecular typing showed that two of three roommates in one room had the same strain, whereas two in another room differed from one another. All isolates, except one, belonged to two related clonal groups. It seems that the clonal group to which most isolates belonged had the greatest potential for spreading among both residents and HCWs. CONCLUSIONS: Similar prevalence rates of MRSA colonization have been found in other European countries, but such studies have usually involved residents with better functional status than that of the participants in this study. Nosocomial spread of MRSA occurred in the facility examined, but not frequently. More attention should be focused on the hand hygiene of HCWs. PMID- 15756891 TI - Risk factors for colonization with methicillin-resistant Staphylococcus aureus in a long-term-care facility in Slovenia. AB - OBJECTIVE: To evaluate risk factors associated with methicillin-resistant Staphylococcus aureus (MRSA) colonization in a long-term-care facility (LTCF) for the elderly in Slovenia. SETTING: A 351-bed community LTCF for the elderly. DESIGN AND PARTICIPANTS: This was a case-control study. MRSA carriage was identified in 102 of 127 residents of the facility's nursing unit. Two swabs were taken: one from the anterior nares and one from the largest skin lesion. If no skin lesions were present, the axillae and the groin area were swabbed. Data were collected regarding gender, age, length of stay in the facility, underlying conditions, functional status, presence of wounds or pressure sores, presence of catheters, antibiotic treatments, and hospital admissions. RESULTS: We detected MRSA in 12 participants. Risk factors independently and significantly associated with MRSA colonization on the multivariate analysis were antibiotic treatments within 1 month before the investigation (odds ratio, 5.087; 95% confidence interval, 1.02 to 25.48; P = .048) and multiple hospital admissions in the 3 months before the investigation (odds ratio, 6.277; 95% confidence interval, 1.31 to 30.05; P = .022). CONCLUSIONS: This is the first assessment of risk factors for colonization with MRSA in an LTCF in Slovenia. MRSA poses a problem in this LTCE Our observations may be valuable in implementing active surveillance cultures in infection control programs in Slovenian LTCFs. PMID- 15756892 TI - Dispersal of Staphylococcus aureus into the air associated with a rhinovirus infection. AB - OBJECTIVE: To determine whether healthy adult nasal carriers of Staphylococcus aureus can disperse S. aureus into the air after rhinovirus infection. DESIGN: We investigated the "cloud" phenomenon among adult nasal carriers of S. aureus experimentally infected with a rhinovirus. Eleven volunteers were studied for 16 days in an airtight chamber wearing street clothes, sterile garb, or sterile garb plus surgical mask; rhinovirus inoculation occurred on day 2. Daily quantitative air, nasal, and skin cultures for S. aureus; cold symptom assessment; and nasal rhinovirus cultures were performed. SETTING: Wake Forest University School of Medicine, Winston-Salem, North Carolina. PARTICIPANTS: Wake Forest University undergraduate or graduate students who had persistent nasal carriage of S. aureus for 4 or 8 weeks. RESULTS: After rhinovirus inoculation, dispersal of S. aureus into the air increased 2-fold with peak increases up to 34-fold. Independent predictors of S. aureus dispersal included the time period after rhinovirus infection and wearing street clothes (P < .05). Wearing barrier garb but not a mask decreased dispersal of S. aureus into the air (P < .05). CONCLUSION: Virus induced dispersal of S. aureus into the air may have an important role in the transmission of S. aureus and other bacteria. PMID- 15756893 TI - Healthcare-associated Staphylococcus aureus bacteremia and the risk for methicillin resistance: is the Centers for Disease Control and Prevention definition for community-acquired bacteremia still appropriate? AB - OBJECTIVE: To evaluate a new classification for bloodstream infections that differentiates hospital acquired, healthcare associated, and community acquired in patients with blood cultures positive for Staphylococcus aureus. DESIGN: Prospective, observational study. SETTING: Three tertiary-care, university affiliated hospitals in Dublin, Ireland, and Strasbourg, France. PATIENTS: Two hundred thirty consecutive patients older than 18 years with blood cultures positive for S. aureus. METHODS: S. aureus bacteremia (SAB) was defined as hospital acquired if the first positive blood culture was performed more than 48 hours after admission. Other SABs were classified as healthcare associated or community acquired according to the definition proposed by Friedman et al. When available, strains of methicillin-resistant Staphylococcus aureus (MRSA) were analyzed by pulsed-field gel electrophoresis (PFGE). RESULTS: Eighty-two patients were considered as having community-acquired bacteremia according to the Centers for Disease Control and Prevention (CDC) classification. Of these 82 patients, 56% (46) had healthcare-associated SAB. MRSA prevalence was similar in patients with hospital-acquired and healthcare-associated SAB (41% vs 33%; P > .05), but significantly lower in the group with community-acquired SAB (11%; P < .03). PFGE of MRSA strains showed that most community-acquired and healthcare-associated MRSA strains were similar to hospital-acquired MRSA strains. On multivariate analysis, Friedman's classification was more effective than the CDC classification for predicting MRSA. CONCLUSION: These results support the call for a new classification for community-acquired bacteremia that would account for healthcare received outside the hospital by patients with SAB. PMID- 15756894 TI - Public disclosure of healthcare-associated infections: the role of the Society for Healthcare Epidemiology of America. PMID- 15756895 TI - An outbreak of methicillin-resistant Staphylococcus aureus surgical-site infections initiated by a healthcare worker with chronic sinusitis. AB - An investigation was performed following two methicillin-resistant Staphylococcus aureus surgical-site infections in a 946-bed French general hospital. The investigation revealed that the outbreak involved 7 patients in 2 surgical wards and that infections were probably contracted in the operating theater from a healthcare worker suffering from chronic sinusitis. PMID- 15756896 TI - Clustering of multiple endemic strains of methicillin-resistant Staphylococcus aureus in a nursing home: an 8-year study. AB - PFGE was performed on residents' first clinical MRSA isolate (n=94) during 8 years. Sixty-one percent of the isolates were clustered in time (P < .05) and space (P < .05) (i.e., 2 separate statistically significant tests). Isolates from individual units were genetically related, with only the occasional unrelated isolate. PMID- 15756897 TI - Dreaming a different epidemiological future. PMID- 15756898 TI - Some challenges for medical statistics. AB - A broad review is given of some of the issues facing medical statistics organized under the headings of measurement, design, analysis and interpretation. PMID- 15756899 TI - Commentaries on 'epidemiology: quo vadis?'. PMID- 15756900 TI - Validity of parental-reported questionnaire data on Danish children's use of asthma-drugs: a comparison with a population-based prescription database. AB - The aim of this study was to examine the validity--estimated as the sensitivity, specificity, and predictive values--of questionnaire-based parental reporting on Danish children's use of asthma drugs. Within a population-based cross-sectional study on the association between asthma and dental anxiety, we compared parental answers on their children's use of asthma drugs during the past 12 months with National Health Service (NHS) data from a prescription database on prescriptions of inhaled beta 2-agonists, inhaled corticosteroids, or both for a period of 14 months preceding the date of posting the parental questionnaires. In four municipalities in North Jutland County, Denmark, in 2001, parents of 1273 children between the ages of 6 and 8 years completed the questionnaire. For frequency of use, the sensitivity increased from 57.9% (95% confidence interval [CI]: 47.8-67.3%) for the answer 'Every day' to 91.6% (95% CI: 84.3-95.7%) when the answers 'Every day', 'Often', and 'Sometimes' were combined. The specificity exceeded 95.9% (95% CI: 94.6-96.9%) for all answers. The positive predictive value (PPV) increased from 64.4% (95% CI: 56.1-72.0%) when the answers 'Every day', 'Often', and 'Sometimes' were combined to 94.8% (95% CI: 85.9-98.2%) for the answer 'Every day'. The negative predictive value (NPV) both exceeded 96.7% (95% CI: 95.5-97.6%) for all answers. Questionnaire-based parental reporting on children's one-year asthma-drug use appears to be valid in epidemiological research depending on the study question and design. PMID- 15756901 TI - Bayes' theorem: a negative example of a RCT on grommets in children with glue ear. AB - Bayesian inference presupposes that practitioners' belief in the effectiveness of medical intervention is the product of prior belief and recent evidence from studies. Although increasingly used, up to now the posterior belief calculated according to the theorem has not been compared with an empirically measured posterior belief. We conducted a RCT, which was preceded by elicitation of prior beliefs among ENT-surgeons, and which was followed by elicitation of posterior beliefs among ENT-surgeons, 1 year after completion of the trial. We compared the posterior beliefs of ENT-surgeons about the effect of grommets in children with glue ears, as predicted by Bayes' theorem with actual measured posterior beliefs. The distribution of the measured posterior beliefs was not in line with the calculated posterior, but almost identical to the distribution of the measured prior beliefs. The results showed that our trial had little or no impact on the beliefs of the ENT-surgeons, i.e. they did not adjust their belief to the extent that was expected according to Bayes' theorem. PMID- 15756902 TI - Generalized additive models applied to analysis of the relation between amount and type of alcohol and all-cause mortality. AB - The J-shaped relation between alcohol intake and mortality is well established, whereas the nadir of the curve is not determined. Due to non-linearity of the relation, categorical alcohol variables have been used to model the relation. In Generalized Additive Models (GAM) non-linear relations can be modelled without the disadvantages of categorization and without assumptions regarding the functional form. The aim of this study was to use GAM to evaluate the relation between alcohol intake, amount and type, and mortality. The relation was investigated using data from the Copenhagen City Heart Study (11,920 participants of whom 5552 died during 20 years follow-up). Using GAM, a smooth J-shaped relation between alcohol and mortality was found. However, if non-drinkers were categorized separately there was a positive association between alcohol and mortality even for low alcohol intake. For equal total alcohol intake, men and women drinking wine or spirits had lower mortality than beer drinkers. The nadir of the relation between alcohol and mortality was sensitive to the handling of non-drinkers. When non-drinkers were categorized separately we found no indication of a beneficial influence of low alcohol intake on mortality. PMID- 15756903 TI - Association of alcohol consumption to mortality and person-years of life lost in Switzerland--measuring the impact of some methodological options. AB - OBJECTIVES: The present study is designed to estimate the health impact of alcohol consumption in terms of the number of deaths and person-years of life lost (PYLL) in Switzerland. To that end, three hypothetical situations have been compared to the current situation used as a reference. The underlying question was: how many deaths and person-years of life lost would be avoided if all individuals were: (1) non-drinkers, (2) low drinkers only or (3) both (either abstainers or low level drinkers)? METHODS: Etiological fractions of alcohol related health consequences by age and gender were applied to the officially reported numbers of deaths to estimate the total number of deaths and PYLL. RESULTS: Depending upon the hypothetical situation, the estimated number of alcohol-related deaths varied between an overall net protective and an overall net causal effect: in situation (1), there would occur 730 additional deaths in reference to the current situation (+1.2% of the total alcohol-related mortality in Switzerland), whereas in situation (2) there would be 3460 deaths less ( 5.5%). Thus, the current situation in Switzerland would be preferable compared to a society, in which no one drinks alcohol. A society, however, in which every individual drinks at low levels, would be preferable to the current situation. The corresponding figures for PYLL are 23,596 vs. 29,229 (6.4% vs. 7.9% of total PYLL). CONCLUSIONS: The hypothetical situation decisively impacts on the number of deaths attributable to alcohol. In contrast, it has much less influence on the number of alcohol-related PYLL. Thus, mortality is at least a questionable indicator of alcohol-related health outcomes. The study also indicated that more lives and PYLL were saved in the low-risk situation than in the abstinence situation. Public health policies should not concentrate on the promotion of abstinence. PMID- 15756904 TI - Contributions of mortality changes by age group and selected causes of death to the increase in Japanese life expectancy at birth from 1950 to 2000. AB - The purpose of this study is to analyze contributions of mortality change by age group and selected causes of death to the increase in life expectancy at birth from 1950 to 2000 in Japan, which has the longest longevity in the world. Using mortality data from Japanese vital statistics from 1950 to 2000, we analyzed contributions of mortality change by age group and selected causes of death to the increase in life expectancy at birth by the method of decomposition of changes and calculated age-adjusted death rates for selected causes of death. Gastroenteritis, tuberculosis and pneumonia largely contributed to an increase in life expectancy in childhood and in the young in the 1950s and 1960s. The largest contributing disease changed from tuberculosis and pneumonia in earlier decades to cerebrovascular diseases in the 1970s. The largest contributing age group also shifted to older age groups. Age-adjusted death rate for cerebrovascular diseases in 2000 was one fifth of the 1965 level. Cerebrovascular diseases contributed to an increase in life expectancy at birth of 2.9 years in males and 3.1 years in females from 1970 to 2000. In the 1990s, the largest contributing age group, both among males and among females, was the 75-84 age group. Of the selected causes of death, heart diseases other than ischemic heart disease became the largest contributor to the increase in life expectancy at birth. Unlike cerebrovascular diseases, cancer and ischemic heart disease contributed little to change in life expectancy at birth over the past 50 years. In conclusion, although mortality from ischemic heart disease has not increased since 1970 and remained low compared with levels in western countries, mortality from cerebrovascular diseases has dramatically decreased since the mid-1960s in Japan. This gave Japan the longest life expectancy at birth in the world. It is necessary to study future trends in life expectancy at birth in Japan. PMID- 15756905 TI - Chlamydia pneumoniae seropositivity and risk of ischemic stroke: a nested case control study. AB - Chlamydia pneumoniae has been linked with increased risk of cardiovascular disease, but data on stroke are sparse. We examined whether seropositivity to Chlamydia pneumoniae was associated with the risk of ischemic stroke in a nested case-control study. Data on Chlamydia pneumoniae serology, lifestyle factors, and medical history were obtained at baseline. Verified cases (n = 254) were compared with gender- and age-matched controls (n = 254). Positive IgA (> or = 1:16) or IgG (> or = 1:64) titers were associated with an increased risk of acute ischemic stroke, i.e. adjusted odds ratios (ORs) were 1.54 (95% confidence interval, CI: 0.96-2.47) and 1.28 (95% CI: 0.83-1.95). The adjusted OR was 1.77 (95% CI: 1.04 3.00) when both titers were elevated. The highest point estimates were seen for ischemic stroke due to large-artery atherosclerosis, adjusted OR: 6.32 (95% CI: 0.76-52.61) (IgG (> or = 1:64)). No clear associations were found for other types of ischemic stroke. The strength of the association varied depending on gender and the chosen cut-off values for the antibody titers. These results partly support the hypothesis that serologic evidence of Chlamydia pneumoniae infection may be associated with an increased risk of ischemic stroke. However, the risk may differ according to gender, subtype of ischemic stroke, and cut-off value of antibody titers. PMID- 15756906 TI - Effect modification by vitamin C on the relation between gastric cancer and Helicobacter pylori. AB - A hospital-based case-control study of 295 cases with histologically confirmed gastric cancer and age and sex-matched controls was conducted to evaluate the effect of dietary vitamin C intake upon the relation between Helicobacter pylori infection and gastric cancer in Korea in 1997-1998. Anti-H. pylori IgG was detected by ELISA. A food frequency questionnaire, and a questionnaire on demographic factors, including past medical history, smoking, alcohol consumption, and life style was also administered. The prevalences of H. pylori IgG in cases and controls were 80.7% and 71.2%, respectively, and the odds ratio (OR) of H. pylori for gastric cancer was 1.68 (95% confidence interval (CI): 1.14, 2.44), after adjusting for age, sex, educational level, and a past medical history of gastritis or gastric ulcer. In a stratified analysis, H. pylori seropositivity was found to be a significant risk factor for gastric cancer in the low vitamin C intake group (OR = 4.68; 95% CI: 1.97, 11.1), but not in the high vitamin C intake group (OR = 0.72; 95% CI: 0.32, 1.65). Vitamin C intake was found to modify the relation between H. pylori and gastric cancer. PMID- 15756907 TI - Confounding by dietary factors in case-control studies on the efficacy of cancer screening in Japan. AB - Evaluation of cancer screening using case-control studies is less valid in comparison to randomized controlled trails, due to the intrusion of a possible self-selection bias in the former. The randomized controlled trial approach, however, may be difficult in developed countries where mass cancer screening programs are already being performed nationally. Accordingly, case-control studies are often performed instead of randomized controlled trials. In case control studies, no reports could be found in the literature using dietary habit, an important influencing factor in carcinogenesis, as an adjusting item. We surveyed nutrition and food intake status through a nutrition survey using the weighing method, and smoking prevalence and alcohol consumption with questionnaires in the general population in northern Japan, in subjects over 30 years of age. We then compared these results among non-participants and participants in cancer screening programs, and evaluated how any differences between the two groups might affect the results of case-control studies. Non participants had a significantly lower intake of vegetables, carotene, vitamin C, and dietary fiber, which are thought to be beneficial factors in the prevention of carcinogenesis. Non-participants had a significantly higher cigarette smoking prevalence than participants. In relation to the intake of vegetables, a difference between participants and non-participants was evident, as proper adjustment for vegetable intake led to an odds ratio closer to the null value. The value of the odds ratio will probably not decrease, but it might increase when other dietary factors or smoking are taken into consideration. PMID- 15756908 TI - Relationship among metabolizing genes, smoking and alcohol used as modifier factors on prostate cancer risk: exploring some gene-gene and gene-environment interactions. AB - BACKGROUND: Prostate cancer (PCa) is one of the most common male cancers, but the burden of this disease shows remarkable worldwide variation. The role of susceptibility low penetrance genes and environmental factors in the etiology of (PCa) is unclear, but may involve, in some cases, multiple alleles at multiple loci and environmental factors. STUDY OBJECTIVES: To assess whether CYP1A1, GSTM1, GSTT1 susceptibility genotypes, smoking status and alcohol consumption factors contribute to PCa risk, gene-gene and gene-environment interactions were analyzed. DESIGN AND PARTICIPANTS: We explored interactions on a multiplicative scale conducting a population-based case-control and a case-only study on 103 incident PCa patients and 132 unrelated controls. MAIN RESULTS: The interaction odds ratios (IOR) for PCa risk were increased in men who had both susceptibility genotypes GST (M1; T1) null and CYP1A1-M1* in a case-control and case-only design (IOR(cc): 1.11; 95% CI: 0.12-10.02; IOR(cc): 6.23; 95%, CI: 0.51-75.89; IOR(co): 2.80; 95% CI: 0.44-17.45 and IORco: 2.65; 95%, CI: 0.30-25.40). No clear evidence for interaction on a multiplicative scale between smoking status, alcohol consumption and genetic polymorphisms in PCa risk was observed. CONCLUSIONS: Our findings suggest that the interaction between genetic polymorphisms in GST (T1; M1) and CYP1A1-M1* would play a significant role as a modifying factor on PCa risk in Chilean people. However, these preliminary exploratory results should be confirmed in a larger study. PMID- 15756909 TI - Validation of a decision model for preventive pharmacological strategies in postmenopausal women. AB - BACKGROUND: Benefits and risks of a combined hormone replacement therapy (HRT) based on randomized clinical trial emerged on various disease endpoints in 2002. The Women's Health Initiative (WHI) provides an important health answer for healthy postmenopausal women, such as do not use combined HRT to prevent chronic disease, because of the elevated risk of coronary artery disease (CHD), stroke and venous thromboembolism. In March 2004, the NIH stopped the drugs in the estrogen-alone trial after finding an increase risk of stroke and no effect, neither an increase or a decrease, on risk of CHD after an average of 7 years in the trial. On the other hand, raloxifene, which does not seem to significantly increase the risk of cardiovascular events and could retain skeletal benefits without stimulating endometrial and breast tissue, requires decision-makers since no current data on these disease clinical endpoints have been published. OBJECTIVE: To construct a multi-disease model based on patient-specific risk factor profiles, and to validate the multi-disease model with several tools of internal and external validities. METHODS: A Markov state model was developed. The risks of these various diseases (including coronary artery disease, stroke, hip fracture and breast cancer) are derived from published hazards proportional models which take into account significant risk factors. Canadian-specific rates and data sources for these transition probabilities are derived from published studies and Canadian Health Statistics. The validation of our model were based on several tools of internal and external validities, such as Canadian life expectancy, population-based incidence rate of diseases, clinical trials and other published life expectancy models. RESULTS: First, presumably, small changes in the lifetime probability of dying support the hypothesis that the disease states operate in a largely independent fashion. For instance, the difference in the probability of dying from a particular disease by the complete elimination of a selected disease, such as CHD, stroke or breast cancer, ranged from 0.2 to 2.2% of difference in the lifetime probability of dying of these diseases. Second, we demonstrated that the model adequately predicted the Canadian population lifetable and disease-incidence rates from population-based data among women from 45 to 75 years old. The predictions of the model were cross-checked from non source data, such as predicted outcomes versus observed outcomes from results of clinical trials. Predicted relative risks of CHD event, breast cancer and hip fracture fell in the reported 95% confidence interval of clinical trials. Finally, predicted treatment benefits are comparable with those of published life expectancy models. CONCLUSIONS: The results of the study demonstrated that this multi-disease model, including coronary artery disease, stroke, hip fracture and breast cancer, is a valid model to predict the impact on life expectancy or number of events prevented for preventive pharmacological interventions. PMID- 15756911 TI - The problem of attrition in a Finnish longitudinal survey on depression. AB - A cohort of all school children aged 16 years in 1983 (n = 2194, 96.7%) in Tampere, Finland were studied at 16, 22 and 32 years of age by self-reported questionnaires. The non-response pattern was considered by modelling the individual response probability by panel year and gender. Gender and school performance at age 16 years were the most important predictors of non-response. They explained away the effect of all other variables at 16 and 22 years, except for earlier non-response at age 22. However, the ability of the models to predict non-respondents was very poor. The effect of attrition for the estimation of depression prevalence was evaluated first by longitudinal weighting methods used commonly in survey studies and then by Markov chain Monte Carlo (MCMC) simulation of the missing depression status. Under the missing-at-random assumption (MAR), both applied correction methods gave estimates of roughly the same size and did not significantly differ from the observed prevalence of depression. No indication of informative missingness was found. We therefore conclude that attrition does not seriously bias the estimation of depression prevalence in the data. In general, non-response models, which are needed to correct for informative missingness, are likely to have poor ability to predict non-response. Therefore, the plausibility of the MAR assumption is important in the presence of attrition. PMID- 15756910 TI - Lifetime prevalence estimates of major depression: an indirect estimation method and a quantification of recall bias. AB - The measurement of lifetime prevalence of depression in cross-sectional surveys is biased by recall problems. We estimated it indirectly for two countries using modelling, and quantified the underestimation in the empirical estimate for one. A microsimulation model was used to generate population-based epidemiological measures of depression. We fitted the model to 1-and 12-month prevalence data from The Netherlands Mental Health Survey and Incidence Study (NEMESIS) and the Australian Adult Mental Health and Wellbeing Survey. The lowest proportion of cases ever having an episode in their life is 30% of men and 40% of women, for both countries. This corresponds to a lifetime prevalence of 20 and 30%, respectively, in a cross-sectional setting (aged 15-65). The NEMESIS data were 38% lower than these estimates. We conclude that modelling enabled us to estimate lifetime prevalence of depression indirectly. This method is useful in the absence of direct measurement, but also showed that direct estimates are underestimated by recall bias and by the cross-sectional setting. PMID- 15756912 TI - Re: "Development of a semi-quantitative Food Frequency Questionnaire (FFQ) to assess dietary intake of multiethnic populations" by Danit Shahar, Iris Shai, Hillel Vardi, Ayelet Brener-Azrad & Drora Fraser. PMID- 15756913 TI - [Laparoscopic cholecystectomy as trendsetter in minimal-invasive surgery]. AB - Only 10 years after the introduction of laparoscopic cholecystectomy in 1986 this method has become the "gold-standard" in the treatment of uncomplicated and complicated gallbladder disease. Laparoscopic cholecystectomy was so successfull, that it became the trendsetter in minimal-invasive surgery leading to revolutionary changes in all fields of surgery. Although nowadays self-evident, minimal-invasive surgery passed through a protracted and hindrance history of development. In this article we will review the history of laparoscopy and laparoscopic cholecystectomy and discuss its influence in the development of minimal-invasive surgery. PMID- 15756914 TI - [Learning curve--calculation and value in laparoscopic surgery]. AB - The learning curve shows the progress in mastering a new method. It is completed when the monitored parameters reach a steady state and when the final results can be compared with literature. The earlier used analysis of the performance improvement with its "on the spots" appraisals at certain time-intervals is replaced by a continuous assessment. The multimode learning curve is particularly useful for it, because not only one parameter (f.e. operation-time), but also several important factors can be put together into one single graphic. For the operation-time, the Moving Average Method is useful. For incidents, which may happen or not like a conversion from laparoscopy to laparotomy as well as complications, the Cusum-method is of practical use. The learning curves of the technique of laparoscopic cholecystectomy, colo-rectal surgery, fundoplicatio and hernia surgery have been completed. Also, the learning curve of the industry is well advanced. Reliable data for the learning curves of individual surgeons for certain operations cannot be given, as, only now, young doctors are being trained on a large scale in laparoscopic technique as used to be the case in the open abdominal surgery. This will influence greatly the learning curves and will shorten the time till their completion. Different bias concerning the individual surgeons and their clinics prohibit the production of comparable curves. Several factors like the patient respectively his abdomen are complicating all this. That's why the learning curves cannot be used as benchmarks to compare different surgeons or clinics, as long as no valid scoring system concerning the complexity of a surgical intervention exists. Learning curves which become quality curves after reaching a steady state, can be used for the individual monitoring of a surgeon's performance and serve as a quality measurement of a clinic. The learning curves of the laparoscopic cholecystectomy, fundoplicatio, colo-rectal surgery and hernia surgery are discussed in particular The mandatory number of operations needed to learn a new method cannot yet be established today, even if all the existing data are consulted. Therefore, the learning curve is a useful instrument to monitor the individual progress and the results of a clinic in the meaning of an individual quality-management. After completion of the learning curve, a quality curve using the same parameters will be given, which shows the deviations of its own standard. PMID- 15756915 TI - [The role of video-thoracoscopy in thoracic surgery]. AB - The widespread acceptance of minimal invasive techniques has revolutionized the practice of surgery including, thoracic surgery. Within a short period of time, video-assisted thoracic surgery (VATS) has become an acceptable approach to a wide range of thoracic procedures. The use of VATS as a diagnostic modality is now well established. For therapeutic procedures, VATS has also been generally accepted as the treatment for spontaneous pneumothorax, thoracic sympathectomy, treatment of loculated effusions and resection of simple mediastinal cysts and benign tumors. Its role in major procedures, e.g. anatomic lung resections and thymectomy, however, remain poorly defined at present although some of the existing intermediate results are encouraging. The technique continues to evolve, with further miniaturization to reduce access-induced trauma. No matter how attractive the new techniques may appear, carefully conducted clinical trials should precede the general acceptance and widespread use. PMID- 15756916 TI - [Endoscopic repair of groin hernia--what ist evidence-based]. AB - The endoscopic techniques of groin hernia repair have developed tremendously over the past 10 years. There are numerous prospective randomized clinical trials comparing conventional techniques of groin hernia repair with endoscopic methods. However, it is difficult to establish a proper meta-analysis of the currently available data due to widely differing study designs. We assessed the currently available prospective randomized clinical trials comparing conventional versus laparoscopic hernia repair. We conclude that based on present data there is a clear advantage for the endoscopic techniques regarding length of hospital stay, postoperative recovery and return to normal activity. The conventional techniques are superior regarding duration of operation and direct cost. Rate of recurrence are comparable in both, endoscopic and conventional approach using mesh prosthesis. Endoscopic techniques are far superior to conventional operation without the use of mesh prosthesis. In patients with recurrent hernia endoscopy offers a major advantage regarding postoperative pain, lenght of hospital stay, return to normal activity and rate of recurrence. We demonstrate these suggestions discussing our own results. Therefore we recomend endoscopic hernia repair in patients with bilateral or recurrent hernia. PMID- 15756917 TI - [Current concepts in minimal invasive endocrine surgery]. AB - During the past ten years, endoscopic adrenalectomy has emerged as the standard of treatment of benign diseases of the adrenal gland. Although no randomised trial exists comparing open with endoscopic adrenalectomy, the endoscopic approach has proven to be safe with lower morbidity and shorter hospital stay compared with open surgery. For parathyroidectomy several different minimal invasive techniques have been described (i.e. fully endoscopic, videoscopic assisted, and focused using mini-incision). Prerequisits for all of these approaches are a positive preoperative localisation of the adenoma, an intraoperative parathormone testing, and the respective surgical experience in the minimal invasive technique. Proponents of the videoendoscopic assisted thyroidectomy mention the enhanced exposure of the surgical field to be beneficial. However, drawbacks of this technique are the fact that a minority of patients qualify for this approach and that the learning curve is quite long for the surgeon. The experience with the laparoscopic approach to endocrine pancreatic tumours is still limited. Good indications are insulinomas that are located anteriorly or within the tail of the pancreatic gland. Nowadays, laparoscopic enucleation of such tumours and tail resections become feasable and safe. PMID- 15756918 TI - [Laparoscopic fundoplication for gastroesophageal reflux disease (GERD)]. AB - Laparoscopic fundoplication became the gold standard in the surgical therapy of GERD. In comparison with open procedures, laparoscopic antireflux surgery has a lower morbidity rate, a better early and late postoperative outcome and is more cost-effective. Antireflux surgery can be performed after a critical evalutation of the patient including gastroscopy, biopsy, 24h-pH-manometry and after a long lasting conservative medical treatment. Indications for antireflux surgery are given by a failed medical treatment, an insufficient compliance, complications of GERD, i.e. stenosis, Barrett-esophagus and atypical reflux symptoms like chronic cough, hoarseness or thoracic pain with presence of a pathological pH-monitoring. Laparoscopic 360 degrees Nissen-fundoplication with crurorrhaphy is our standard procedure, whereas the 270 degrees Toupet technique in our tailored approach is the technique of choice for esophageal motility disorders. Results of antireflux surgery published in literature are discussed and compared with our own ten years experience with 124 cases of laparoscopic fundoplication. PMID- 15756919 TI - [Minimally invasive surgery for morbid obesity--evidence or fashion?]. AB - In the last decades surgical treatment for morbid obesity has emerged from being the interest of only few surgeons to a well recognized surgical specialty. This development was promoted by the dramatic increase of prevalence of obesity and the development of laparoscopic surgical techniques. In 1996 the NIH consensus conference stated that bariatric surgery is the most effective therapy to treat obesity and type 2 diabetes, whereas conservative treatment strategies failed in the long-term. Only few studies have compared the different operative strategies. It has been shown that the bypass procedure is better than gastric banding with respect to weight loss and the decrease of comorbidities, but suffers from more short-term morbidity. In recent years, randomized trials have been published comparing laparoscopic with open procedures. These studies show an advantage for the laparoscopic techniques. This paper gives a critical overview on bariatric surgery and summarizes the current literature in this speciality. PMID- 15756920 TI - [Minimally-invasive management of common bile duct stones]. AB - Common bile duct stones may present a health hazard for our patients. Nevertheless, since the implementation of laparoscopic cholecystectomy optimal diagnostic and therapeutic algorithm are not yet defined. Symptomatic calculi can be assumed on the basis of pathological laboratory values or diagnosed by means of ultrasound, Intraoperative Cholangiography (IOC) or Magnetic-Resonance Cholangio-Tomography (MRCT). For therapy of common bile duct stones endoscopic and laparoscopic minimally-invasive strategies are available. As any type of management may show some benefit, it is not yet evident which policy we should prefer. Specialists do not agree on the necessity of therapy in asymptomatic patients with common bile duct calculi at all. This article shows a current state of the opinion and art and tends to highlight trends and future perspectives. PMID- 15756921 TI - [Is open appendectomy still up-to-date?]. AB - The acceptance of the laparoscopic technique for appendectomy can't be compared with the success in laparoscopic cholecystectomy. The discussion is still controversial. A lower rate of wound infection in comparison to open appendectomy has been proven statistically in many studies. There is no evidence of a higher rate of complications. Postoperative pain may be reduced and return to work is often earlier. Cosmesis shows better results. Operation time and hospital stay are comparable to open appendectomy. The total of direct and indirect costs is not higher with laparoscopic than with open appendectomy. Excellent indications for laparoscopic appendectomy are atypical pain in the right lower abdomen, uncertain diagnosis in fertile and elderly women, obesity and complicated appendicitis. Controlled double-blinded randomized studies are necessary in the future to define the advantages and further indications. PMID- 15756922 TI - [Indications for laparoscopic colorectal resections--also for cancers?]. AB - In the last decade, laparoscopy has dramatically changed colonic surgery. Laparoscopic procedures are applied to the treatment of almost all colonic diseases, including both benign and malignant lesions. Significant benefits can be expected with a laparoscopic approach relative to decreased pain, ileus, length of hospital stay, disability, and possibly, adhesion formation and subsequent bowel obstruction, and improved cosmesis. However, all those benefits are secondary in the treatment of cancer; tumor-free survival must be the primary goal. Early comparative studies have shown that laparoscopic colectomy for cancer is safe, feasible, and an oncologic adequate resection can be performed with acceptable operative times and conversion rates. The most recent results of prospective randomized trials are now showing similar survival and cure rates after laparoscopic colectomy. There is evidence that laparoscopic colectomy for curable cancer results in equivalent cancer related survival compared to open colectomy when performed by experienced surgeons. The surgical techniques are demanding and require a level of standardization to achieve success. Laparoscopic colorectal surgery for cancer will have a definite role in the future. PMID- 15756923 TI - [Minimal invasive surgery in gynaecology]. AB - In Gynaecology there have been many innovative developments towards minimal invasive surgery. Today patients can profit of these new techniques in all fields of gynaecological surgery, some of which are described: The treatment of benign adnexal pathology including ectopic pregnancy is a domain of laparoscopy. Also symptomatic myomas can be enucleated by laparoscopy with favourable pregnancy outcome. The role of endoscopy in malignant disease has still to be defined though pelvic and paraaortal lymphonodectomy and even radical hysterectomy have been shown to be feasible. Pathology of the uterine cavity (myomas, polyps, septae) is routinely treated by hysteroscopy. The concept of operative treatment of stress urinary incontinence has changed since the minimal invasive TVT procedure has shown to be as successful as the classic colposuspension. For patients with breast cancer there has been a change from (ultra-) radical surgery to breast conserving treatment. Furthermore with the concept of the sentinel lymphnode a reduction of the morbidity of the classical lymphonodectomy is achieved without loss of information about the axillary lymphnode status. PMID- 15756924 TI - [Arthroscopic surgery]. AB - Although arthroscopy of the knee joint had already been reported during the 1930's, the general dissemination of this method first began in the 1970's. The main reason for the rapid dissemination of this method was especially the fact that in addition to diagnostics, therapeutic possibilities were recognized and immediately implemented. This meant that arthroscopy had great potential and was made well known since the surgery was minimally invasive. Today we can assume that the technological side of the arthroscopic method is very widely developed and new innovations only arise slowly. Innovations are mostly connected with new innovative operating techniques. Surgery of the knee joint was the dominant application of arthroscopy in the beginning. The method was quickly applied to other joints. Today there is practically no joint which is inaccessible to arthroscopy. From surgical and therapeutic perspectives, arthroscopy is most frequently used today for the knee joint, followed by the shoulder joint, ankle joint, elbow joint, hip joint as well as wrist joint. Arthroscopic surgery within the field of joint surgery is regarded as indispensable. This specific surgery needs corresponding ability and skill, which must be individually acquired. Arthroscopic surgery evidently depends on technology and accordingly requires a corresponding fully operational medical infrastructure and knowledge. The big advantage of arthroscopic surgery lies in the minimally invasive technique, which has reduced the primary postoperative mortality significantly. Therefore, with good indicators the patient has decisive advantages as well as good cost to benefit ratios. PMID- 15756925 TI - [Actual relevance of minimal invasive surgery in fracture treatment]. AB - Minimal invasive osteosynthesis (MIO) should belong nowadays in the armentarium of each trauma surgeon. The tendency to minimize the invasivity of every operation is a logical development considering the goal of each surgeon to reduce the iatrogenic damage caused by the operation. The term 'MIO" stands for this criteria: Fracture zone not opened, reduction by indirect manoeuvres or percutaneously by joy-sticks, small approaches for the application of the implants, intraoperative assessment of the reduction by imaging (fluoroscopy, arthroscopy, endoscopy, etc.). Long time before the definition of "minimal invasive surgery" some technologies in trauma surgery as external fixator or intramedullary nailing already fulfilled these criteria. In the nineties of the 20th century first cases of percutaneously inserted plate osteosynthesis have been reported. This technique got a widespread acceptance under the term of MIPO (minimal invasive plate osteosynthesis) during the last five years, especially pushed with the new angular stable screw-plate systems (LISS, LCP). The main problem of the MIPO-technique is and remains the reduction (no direct manipulation possible) and their intraoperative assessment (no direct visualisation). The balance between the degree of invasivity and the achieved quality of reduction and stability is often difficult to define and must be related to several factors (localisation and type of fracture, local soft tissue conditions, quality of the bone, age and wishes of the patient, available implants, experience of the surgeon, etc.). New technologies as improved imaging, intraoperative navigation and percutaneous reduction tools will help to further reduce the invasivity of fracture surgery in the future. PMID- 15756926 TI - Repeated restraint stress reduces opioid receptor binding in different rat CNS structures. AB - Different effects of exposure to acute or to repeated stress have been observed upon the nociceptive response in rats. In the present study, we repeatedly submitted Wistar rats to restraint for 40 days, a treatment known to induce an increase in the nociceptive response in the tail-flick test. Afterwards, the effect of repeated restraint stress on the density of opioid receptors in rat spinal cord, frontal cortex, and hippocampus was investigated. Results showed that repeatedly stressed rats displayed a significant decrease in opioid receptors density in all structures studied; cortex (141.3 +/- 5.7 for control and 103.3 +/- 15.9 for stressed rats), hippocampus (92.4 +/- 7.2 for control and 64.8 +/- 7.7 for stressed rats), and spinal cord (122.2 +/- 12.8 for control and 79.7 +/- 9.7 for stressed rats). These findings suggest opioid mediation of the altered responses observed in these repeatedly-stressed animals, although the participation of non-opioid mechanisms in this phenomenon cannot be ruled out. PMID- 15756927 TI - Expression of N-myristoyltransferase in human brain tumors. AB - N-myristoylation is a process of covalent irreversible protein modification that promotes association of proteins with membranes. Based on our previous findings of elevated N-myristoyltransferase (NMT) activity in colonic epithelial neoplasms that appears at an early stage in colonic carcinogenesis, together with elevated NMT expression in human colorectal and gallbladder carcinomas, we investigated NMT activity and protein expression of NMT1 and NMT2 in human brain tumors and documented elevated NMT activity and higher protein expressions. For the first time, we have demonstrated that NMT has the potential to be used as a marker of human brain tumors. However, further studies with larger number of patients are required to establish its role as a complementary diagnostic tool. This finding has significant implications for further understanding of biological mechanisms involved in tumorigenesis, as well as for diagnosis and therapy of human brain tumors. PMID- 15756928 TI - Cytidine 5'-diphosphocholine (CDP-choline) in stroke and other CNS disorders. AB - Brain phosphatidylcholine (PC) levels are regulated by a balance between synthesis and hydrolysis. Pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1alpha/beta) activate phospholipase A(2) (PLA(2)) and PC-phospholipase C (PC-PLC) to hydrolyze PC. PC hydrolysis by PLA(2) releases free fatty acids including arachidonic acid, and lyso-PC, an inhibitor of CTP-phosphocholine cytidylyltransferase (CCT). Arachidonic acid metabolism by cyclooxygenases/lipoxygenases is a significant source of reactive oxygen species. CDP-choline might increase the PC levels by attenuating PLA(2) stimulation and loss of CCT activity. TNF-alpha also stimulates proteolysis of CCT. TNF-alpha and IL-1beta are induced in brain ischemia and may disrupt PC homeostasis by increasing its hydrolysis (increase PLA(2) and PC-PLC activities) and inhibiting its synthesis (decrease CCT activity). The beneficial effects of CDP-choline may result by counteracting TNF alpha and/or IL-1 mediated events, integrating cytokine biology and lipid metabolism. Re-evaluation of CDP-choline phase III stroke clinical trial data is encouraging and future trails are warranted. CDP-choline is non-xenobiotic, safe, well tolerated, and can be considered as one of the agents in multi-drug treatment of stroke. PMID- 15756929 TI - Decreased 5-HT1A receptor gene expression and 5-HT1A receptor protein in the cerebral cortex and brain stem during pancreatic regeneration in rats. AB - The present study was to investigate the role of central 5-HT and 5-HT(1A) receptor binding and gene expression in a rat model of pancreatic regeneration using 60% pancreatectomy. The pancreatic regeneration was evaluated by 5-HT content and 5-HT(1A) receptor gene expression in the cerebral cortex (CC) and brain stem (BS) of sham operated, 72 h and 7 days pancreatectomised rats. 5-HT content significantly increased in the CC (P < 0.01) and BS (P < 0.05) of 72 h pancreatectomised rats. Sympathetic activity was decreased as indicated by the significantly decreased norepinephrine (NE) and epinephrine (EPI) level (P < 0.001 and P < 0.05) in the plasma of 72 h pancreatectomised rats. 5-HT(1A) receptor density and affinity was decreased in the CC (P < 0.01) and BS (P < 0.01). These changes correlated with a diminished 5-HT(1A) receptor mRNA expression in the brain regions studied. Our results suggest that the brain 5-HT through 5-HT(1A) receptor has a functional role in the pancreatic regeneration through the sympathetic regulation. PMID- 15756930 TI - The oxidative and morphological effects of high concentration chronic toluene exposure on rat sciatic nerves. AB - This study was designed to investigate the effects of chronic toluene inhalation in high concentration on lipid peroxidation, antioxidant enzyme activities and ultrastructural changes in the sciatic nerves of rats. Male Wistar albino rats (150-250 g) were divided in two experimental groups: the control and the toluene treated group (n = 10 for each). Toluene treatment was performed by inhalation of 3000 ppm toluene, in a 8 h/day and 6 day/week order for 16 weeks. Blood and tissue samples were obtained for biochemical and histopathological investigation. The blood and sciatic nerves were assayed for toluene by gas chromatography. Toluene significantly increased blood and tissue malondialdehyde (MDA), and decreased tissue superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), but not tissue catalase (CAT) levels when compared with controls. Electron micrographs of sciatic nerve in the toluene group shows myelin destructions with onion-bulb and bubble form protrusion on the myelin sheath and axolemma border of myelinated axons. The area of injury on the myelin sheath were measured by Image Pro Plus. Mean of the injury area were estimated 34% each myelin. These findings indicate that chronic toluene inhalation might be involved with free radical processes. PMID- 15756931 TI - Tenoxicam exerts a neuroprotective action after cerebral ischemia in rats. AB - In this study we investigated the effects of Tenoxicam, a type 2 cyclooxygenase (COX-2) inhibitor, on brain damage induced by ischemia-reperfusion. Male Wistar rats (18-month old average) were anesthetized and submitted to ischemia occlusion of both common carotid arteries (BCAO) for 45 min. After 24 h of reperfusion, rats were decapitated and hippocampi removed for further assays. Animals were divided into sham-operated, ischemia, ischemia + Tenoxicam 2.5 mg/kg, and ischemia + Tenoxicam 10 mg/kg groups. Tenoxicam was administered intraperitoneally immediately after BCAO. Histological analyses show that ischemia produced significant striatal as well as hippocampal lesions which were reversed by the Tenoxicam treatment. Tenoxicam also significantly reduced, to control levels, the increased myeloperoxidase activity in hippocampus homogenates observed after ischemia. However, nitrite concentrations showed only a tendency to decrease in the ischemia + Tenoxicam groups, as compared to that of ischemia alone. On the other hand, hippocampal glutamate and aspartate levels were not altered by Tenoxicam. In conclusion, we showed that ischemia is certainly related to inflammation and to increased free radical production, and selective COX-2 inhibitors might be neuroprotective agents of potential benefit in the treatment of cerebral brain ischemia. PMID- 15756932 TI - Administration of myo-inositol plus ethanolamine elevates phosphatidylethanolamine plasmalogen in the rat cerebellum. AB - Plasmalogens are ether-linked phospholipids highly abundant in nervous tissue. Previously we demonstrated that acute administration of myo-inositol (myo-Ins) + [2-(13)C] ethanolamine ([2-(13)C]Etn) significantly elevated phosphatidylethanolamine plasmalogen (PlsEtn) in rat whole brain. Current experiments investigated the effects of acute myo-Ins+[2-(13)C]Etn administration on [PlsEtn] and the biosynthesis of new Etn lipids using NMR spectroscopy in rat cerebral cortex, hippocampus, brainstem, midbrain and cerebellum. Treated rats received a single dose of myo-Ins + [2-(13)C]Etn and controls received saline rather than myoIns. Data reveal that the cerebellum is the brain region most affected by treatment, which resulted in a 22% increase in [PlsEtn] and 89% increase in newly synthesized Etn lipids relative to controls (P < 0.05). Furthermore, the cerebellar PlsEtn/phosphatidylethanolamine ratio and molar percentage of PlsEtn were significantly elevated by 12% and 8%, respectively (P < 0.05). These data suggest that myo-Ins influences Etn lipid metabolism in brain, particularly in the cerebellum where there is a stimulation in the biosynthesis of new Etn lipids with a preference towards PlsEtn. PMID- 15756933 TI - Pretraining but not preexposure to the task apparatus prevents the memory impairment induced by blockade of protein synthesis, PKA or MAP kinase in rats. AB - Adult male Wistar rats were trained and tested in a step-down inhibitory avoidance task (0.4 mA footshock, 24 h training-test interval). Fifteen minutes before or 0, 1.5 or 3 hours after training, animals received a 0.8 microl intrahippocampal infusion of the protein synthesis inhibitor anisomycin (80 microg), the PKA inhibitor Rp-cAMP (0.05 microg), the MAPK kinase inhibitor PD 098059 (50 microM solution) or vehicle (phosphate buffer in saline, pH 7.4). Anisomycin, Rp-cAMP and PD 098059 impaired retention test performance in animals injected at different times, prior and after training. Pretraining with a low footshock intensity (0.2 mA) 24 h before training prevented the amnestic effect of all drugs studied. However, simple preexposure to the inhibitory avoidance apparatus did not alter the amnestic effects of all drugs. The results suggest that memory processing requires hippocampal mechanisms dependent on protein synthesis, PKA and MAPK kinase at different times after training. These findings suggest that weak training must be sufficient to produce some lasting cellular expression of the experience so that the enhancement of consolidation of a previously acquired memory is not dependent on protein synthesis, PKA or MAPK. PMID- 15756934 TI - Intracerebroventricular guanine-based purines protect against seizures induced by quinolinic acid in mice. AB - Acute and chronic administration of the nucleoside guanosine have been shown to prevent quinolinic acid (QA) and alpha-dendrotoxin-induced seizures, as well as to impair memory and anxiety in rats and mice. In this study, we investigated the effect of i.c.v. administration of guanine-based purines (GTP, GDP, GMP, and guanosine) against seizures induced by the NMDA agonist and glutamate releaser quinolinic acid in mice. We also aimed to study the effects of the poorly hydrolysable analogs of GTP (GppNHp and GTPgammaS) and GDP (GDPbetaS) in this seizure model. QA produced seizures in 100% of mice, an effect partially prevented by guanine-based purines. In contrast to GTP (480 nmol), GDP (320-640 nmol), GMP (320-480 nmol) and guanosine (300-400 nmol), the poorly hydrolysable analogs of GTP and GDP did not affect QA-induced seizures. Thus, the protective effects of guanine nucleotides seem to be due to their conversion to guanosine. Altogether, these findings suggest a potential role of guanine-based purines for treating diseases involving glutamatergic excitotoxicity. PMID- 15756935 TI - Age-related decline of brain monoamines in mice is reversed to young level by Japanese herbal medicine. AB - Young (3-month-old) and aged mice (18-month-old) were fed a diet containing Japanese herbal medicine (TJ-41 or TJ-48) for 5 months, and the effect of the herbal medicines were examined in terms of levels of monoamines and their metabolites in the brain of young and aged mice. In the aged mice, the levels of norepinepherine, serotonin and their metabolites in the brain were decreased in the cortex, hippocampus and hypothalamus. Feeding of diet containing TJ-48, but not TJ-41, enhanced the levels of some monoamines and their metabolites in the brains of aged mice, comparable to those of young mice. The results indicated that the improvement of levels of monoamines by Japanese herbal medicine was observed only in the aged mice, not in the young mice. The data have suggested the importance of the aged animals to see the effect of medicine on the functions of organs or systems. PMID- 15756936 TI - Neurotoxicity induced by glutamate in glucose-deprived rat hippocampal slices is prevented by GMP. AB - Guanosine-5'-monophosphate (GMP) was evaluated as a neuroprotective agent against the damage induced by glutamate in rat hippocampal slices submitted to glucose deprivation. In slices maintained under physiological conditions, glutamate (0.01 to 10 mM), Kainate, alpha-amino-3-hydroxi-5-methylisoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), 1S,3R-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), or L-2-amino-4-phosphonobutanoic acid (L-AP4) (100 microM) did not alter cell membrane permeability, as evaluated by lactate dehydrogenase (LDH) release assay. In slices submitted to glucose deprivation, GMP (from 0.5 mM) prevented LDH leakage and the loss of cell viability induced by 10 mM glutamate. LDH leakage induced by Kainate, AMPA, NMDA or 1S,3R-ACPD was fully prevented by 1 mM GMP. However, glutamate uptake was not altered in slices submitted to glucose deprivation and glutamate analogues. Glucose deprivation induced a significant decrease in ATP levels which was unchanged by addition of glutamate or GMP. Our results show that glucose deprivation decreases the energetic charge of cells, making hippocampal slices more susceptible to excitotoxicity and point to GMP as a neuroprotective agent acting as a glutamatergic antagonist. PMID- 15756937 TI - Regional heterogeneity of nicotine effects on neurotransmitters in rat brains in vivo at low doses. AB - In our recent studies on nicotine-induced changes in neurotransmitters in brain areas associated with cognitive function using a nicotine dose of 0.5 mg/kg administered subcutaneously to conscious freely moving rats, we found changes in dopamine, norepinephrine, and serotonin, and their metabolites, in the areas examined. For the present report we examined changes in these neurotransmitters following administration of lower nicotine doses, to test regional differences in nicotine response and possible threshold levels for some effects of nicotine. The doses used were 0.15 mg/kg and 0.03 mg/kg nicotine administered subcutaneously. Nicotine levels in the brain reached peak values in less than 10 min and decreased with a half-life of about 60 min (0.15 mg/kg) or 30 min (0.03 mg/kg) to values below detection limits (1 ng/g), by the later time points of the 0.03 mg/kg experiments. Nicotine-induced dopamine (DA) increase (and increase in DA metabolites) and decrease in 5-HT levels at 0.15 mg/kg were significant in the cortex, less so in the hippocampus. Norepinephrine (NE) increase at 0.15 mg/ kg was much less significant than found previously at 0.5 mg/kg. At a low nicotine dose (0.03 mg/kg), the significant changes observed were a decrease in 5-HT in the hippocampus and small increases of DA and NE in the prefrontal cortex and of NE in the medial temporal cortex. In the nucleus accumbens DA, NE, and 5-HT and their metabolites in the ventral tegmental area, mostly DA and metabolites were increased. We conclude that in areas of cognitive function nicotine-induced DA changes are more concentration dependent than changes in NE or 5-HT, and that there are regional differences in neurotransmitter changes induced by nicotine, with NE changes detectable only in the cortex and 5-HT changes only in the hippocampus at a low nicotine dose, indicating significant regional variation in sensitivity to nicotine-induced neurotransmitter changes in brain areas associated with cognitive function. The decrease in 5-HT shows that nicotine also has indirect effects caused by neurotransmitters released by nicotine. The effects of a low nicotine dose are more significant in areas of reward function, indicating differences in sensitivity between cognitive and reward functions. PMID- 15756938 TI - Repeated restraint stress induces oxidative damage in rat hippocampus. AB - It has been shown that emotional stress may induce oxidative damage, and considerably change the balance between pro-oxidant and antioxidant factors in the brain. The aim of this study was to verify the effect of repeated restraint stress (RRS; 1 h/day during 40 days) on several parameters of oxidative stress in the hippocampus of adult Wistar rats. We evaluated the lipid peroxide levels (assessed by TBARS levels), the production of free radicals (evaluated by the DCF test), the total radical-trapping potential (TRAP) and the total antioxidant reactivity (TAR) levels, and antioxidant enzyme activities (SOD, GPx and CAT) in hippocampus of rats. The results showed that RRS induced an increase in TBARS levels and in GPx activity, while TAR was reduced. We concluded that RRS induces oxidative stress in the rat hippocampus, and that these alterations may contribute to the deleterious effects observed after prolonged stress. PMID- 15756940 TI - Cerebrospinal fluid taurine after traumatic brain injury. AB - In the experimental setting, taurine is known to be released from swollen cells to reestablish their normal volume. However, its clinical relevance has not been fully understood. This study was undertaken to reveal changes in cerebrospinal fluid (CSF) amino acids concentration in patients with severe traumatic brain injury (TBI). The study included eight patients, in whom a ventricular catheter was inserted to measure intracranial pressure and obtain CSF samples for 5 days. CSF obtained from patients with normal pressure hydrocephalus served as a control. CSF taurine concentration increased 1.8 times control (P < 0.05) after TBI and returned to control value approximately 67 h after injury. Taurine decreased further and remained lower than control thereafter. Phosphoethanolamine showed similar increase, whereas glutamine decreased transiently and arginine remained close to control value. The present data support the period of astrocytic swelling observed after TBI in other morphological studies. The mechanism and consequences of CSF taurine decrease in the subacute stage of TBI need to be elucidated. PMID- 15756939 TI - Quantitative proteomic analysis of mitochondria from primary neuron cultures treated with amyloid beta peptide. AB - Increasing evidence supports a role for altered mitochondrial function in the pathogenesis of neuron degeneration in Alzheimer's disease (AD). Although several studies have examined the effect of amyloid beta peptide (Abeta), on activities of individual proteins in primary neuron cultures, there have been no studies of the effects of Abeta on the mitochondrial proteome. Here, we quantitatively measured changes in mitochondrial proteins of primary rat cortical neuron cultures exposed to 25 microM Abeta(25-35) for 16 h using isotope coded affinity tag (ICAT) labeling and 2-dimensional liquid chromatography/tandem mass spectrometry (2D-LC/MS/MS) which allows simultaneous identification and quantification of cysteine-containing proteins. The analysis of enriched mitochondrial fractions identified 10 proteins including sodium/potassium transporting ATPase, cofilin, dihydropyrimidinase, pyruvate kinase and voltage dependent anion channel 1 that were statistically significantly (P < 0.05) altered in Abeta-treated cultures. Elevations of proteins associated with energy production suggest that cells undergoing Abeta-mediated apoptosis increase synthesis of proteins essential for ATP production and efflux in an attempt to maintain metabolic function. PMID- 15756941 TI - Dual effect of isoprostanes on the release of [3H]D-aspartate from isolated bovine retinae: role of arachidonic acid metabolites. AB - The effect of 8-isoprostanes on potassium (K+)-depolarization-evoked release of [3H]D-aspartate from bovine isolated retinae was investigated. Isolated bovine retinae were prepared for studies of K(+)-evoked release of [3H]D-aspartate using the Superfusion Method. Low concentrations of 8-isoPGF(2alpha) (1-100 nM) inhibited whereas higher concentrations of this 8-isoprostane (100 nM-30 microM) enhanced K(+)-induced [3H]D-aspartate overflow. The excitatory effect of 8 isoPGF(2alpha) was mimicked by thromboxane receptor agonist, U-46619 and blocked by thromboxane receptor antagonist, SQ 29,548 (10 microM). Pretreatment of tissues with the cyclooxygenase (COX) inhibitor, flurbiprofen unmasked an inhibitory effect of high concentrations of 8-isoPGF(2alpha) (1-30 microM) on [3H]D-aspartate release that was attenuated by AH 6809 (10 microM). In conclusion, 8-isoPGF(2alpha) exhibits a dual regulatory effect on K(+)-induced [3H]D-aspartate release in isolated bovine retinae. The inhibitory action caused by 8-isoPGF(2alpha) is due to the activation of EP1/EP2 receptors while the excitatory effects are due to the activation of thromboxane receptors. PMID- 15756942 TI - Evidence of nitrosative damage in the brain white matter of patients with multiple sclerosis. AB - Nitric oxide (NO) has been implicated in the pathophysiology of both experimental autoimmune encephalomyelitis and multiple sclerosis (MS). NO-mediated protein damage in MS appears to be confined to large plaques where 3-nitrotyrosine has been detected. To determine whether nitrosative damage takes place beyond visible MS plaques, the occurrence of various NO-triggered protein modifications in normal-appearing white matter (NAWM) of eight MS brains was assessed and compared to that in white matter (WM) of four control brains. As determined by amino acid analysis and western blotting, no evidence of tyrosine nitration was found in the MS samples studied, suggesting that they did not contain appreciable amounts of plaque-derived material. The amino acid composition of total myelin proteins and proteolipid protein (PLP) was also unaltered in the diseased tissue, as was the fatty acid composition of PLP. In addition, we detected no changes in the number of protein free thiols suggesting that oxidation do not occur to any appreciable extent. However, the levels of nitrite in MS-NAWM were higher than those in control WM, while in the MS-gray matter (GM) the concentration of this ion was unaltered. Furthermore, five of the MS samples analyzed, and the same as those with high levels of glial fibrilary acidic protein, showed increased amounts of protein nitrosothiols as determined by the "biotin switch" method. S-nitrosation of GM proteins was again normal. There was no indication of N-nitrosation of tryptophan and N-terminal amino groups in both control and MS tissue. Overall, the data suggests that WM, but not GM, from MS brains is subjected to considerable nitrosative stress. This is the first report to present direct evidence of increased protein S-nitrosation and nitrite content in the brain parenchyma of MS patients. PMID- 15756944 TI - Something of a paradox. PMID- 15756943 TI - Charge heterogeneity of bovine brain macrophage migration inhibitory factor. AB - Macrophage migration inhibitory factor (MIF) is known as a ubiquitous pluripotent cytokine originally identified for its capacity to inhibit the random migration of macrophages in vitro. It is recognized as an important regulator of the immunological, neuroendocrine and enzymatic processes. MIF is widely expressed in brain, but its role in the nervous system is not yet understood. In the course of the study of the primary structure of bovine brain MIF we have previously identified a number of MIF-related proteins having identical N-terminal sequences. In this paper we report the results of isoelectric focusing of MIF isolated to a homogeneous state from bovine brain that revealed MIF charge heterogeneity. We have detected isoelectric forms of MIF with pI values of 6.9, 7.0, 7.3, and 7.8. The diverse actions of MIF within the immuno-neuroendocrine system is suggested to be a result of its occurrence in different isoforms and oligomerization states. PMID- 15756945 TI - Generation of functional Cerec 3D occlusal surfaces: a comparison of two production methods relevant in practice. AB - The introduction of the Cerec 3D software (R 1500) promised the generation of functional occlusion points and proximal contacts on the basis of a 3-dimensional virtual model on the screen. The objective of this in vitro study was to check this claim in the simulated chairside production of a single tooth restoration. Furthermore, the differences both in conventional finishing and in ceramic glazing were to be determined. For the operator, the latter should simplify or shorten the intraoral finishing and polishing of the occlusal surface, or make it superfluous. To check the suitability of the two methods, these were compared with regard to their quality and the time required to perform them. Ten cavities on articulated models, representing real clinical cases, were each provided with two restorations. One was produced with the glazing method (group A), the other conventionally (group B). The Cerec 3D software, version R 1500, was used for designing the restoration. The restorations made from Vita Mark II ceramic blocks were finished with fine diamond instruments and polished with Sof-Lex disks when using the conventional method. In the alternative method, finishing was practically replaced by glazing the surface. The restorations were evaluated in the articulator, considering both the vertical relation at the incisal pin as well as the accurate position of the contact points. The study showed that functional rehabilitation was possible and no significant qualitative differences occurred in the methods. In this in vitro study, it was found that the glazing method reduced the time required by the operator by around 20% and indicates an improvement in quality. The results of this study promise simplification when the glazing method is used clinically. PMID- 15756946 TI - The learning unit "Orthodontic set-up" as a new-media module in teaching. AB - The present study examines the extent to which computer-assisted learning units provided independently of place and time are used in self-study as a supplement to the classical classroom instruction of dental students. Indications as to whether such teaching modules improve training in orthodontics should be obtained from this. Attention was focussed on the implementation and evaluation of the "Orthodontic set-up" teaching module, which can be accessed in the Internet and Intranet of the university. The didactic arrangement offered classical university courses in parallel (four lectures on the subjects of occlusion, function, diagnostics, and therapy) in addition to the electronically communicated teaching contents. In addition, intensive supervision during the production of the set-up was guaranteed. The use of this multimedia learning concept was in general assessed positively by 63 surveyed students in the 2002/03 winter semester. The results revealed on the one hand the intensity of use and features of the acquisition of knowledge (use types), and on the other hand, in terms of professional relevance, the contents were found to be well explained, didactically attractive, and understandably presented. However, numerous drawbacks were also mentioned (technical and time problems; qualification deficits). The experience gained in this project should encourage more future investment in the development of alternative university didactic models. PMID- 15756948 TI - Anterior rehabilitation using Cerec 3 veneer software--a case report. PMID- 15756949 TI - Chairside correlation in Cerec 3D software. PMID- 15756947 TI - The concept of a clinical round as a virtual, interactive web-based, e-learning model for interdisciplinary teaching. AB - The demonstration of patient case reports in the course of a clinical round is an essential part of teaching medicine and dentistry. However, suitable live patients with particular problems are not always available at a time when teaching is taking place. This project therefore had the objective of establishing a web-based, virtual e-learning concept for demonstrating case reports independent of time and place, with the possibility of an interactive examination, diagnosis, and interdisciplinary therapy decision making for medical and dental students. Anonymized case reports of diseases in the oral and maxillofacial region and the interdisciplinary treatment were digitized and prepared in a web-based format. The technical aspect was based on connecting flash modules with videos and animation, and monitoring through HTML and Javascript. Due to the modular concept and the programming used, the learning environment was independent of platform and open. Independent formats (.swf, .avi, .mpeg, etc.) were integrated into the individual modules. According to a hierarchic decision system, the user was guided interactively to the diagnosis through a differential diagnostic exclusion process. Sound was digitized and integrated in mp3 compressed form in the 3D models for lip-synchronous speech output. The speech output was connected with a virtual 3D tutor that acted in an advisory capacity in reaching a diagnosis and determining therapy. Further sources of information and literature with abstracts or pdf files of the subject related publications were inserted to ensure that the teaching was objective. To conclude the virtual clinical round, a check on learning success was conducted in the form of a multimedia multiple choice test. PMID- 15756950 TI - The innovative DeguDent all-ceramic system: benchmark for zirconia processing. PMID- 15756951 TI - Shading Vita YZ substructures: influence on value and chroma, part I. AB - All-ceramic restorations should reproduce as well as possible the color of the natural teeth to create a restoration in accordance with the esthetic wishes and ideas of the patient. The basic color of zirconium oxide is white to ivory. The color can be partially adapted by veneering it with ceramic materials. However, it would be better if the substructure could already be adapted to the basic color shade of the neighboring teeth. In this study, the influence of differently shaded frameworks made of Y-TZP by Vita and 3M ESPE and by Enrico Steger (without, with 0.5 mm, or with 1.0 mm ceramic veneer with Base Dentin) on the brightness, saturation, and color shade (= value, chroma, and hue) was measured and assessed with the aid of a spectral photometer (EasyShade, Vita) in a clinical case in the mouth and on the model. By adaptation to the basic shade, the shading of substructures made of Y-TZP can help to reduce the necessary layer thickness of the veneer ceramic to achieve the desired color and should be performed in the future as a matter of routine. In this way, more substance can be conserved when restoring the teeth without having to accept an impairment of the esthetic result. PMID- 15756952 TI - Science in search for visible in the realm of indiscernable. Dedicated to Gerald Stranzinger. PMID- 15756953 TI - [Surgical treatment of hepatic metastases of colorectal neoplasms]. PMID- 15756954 TI - [Surgical treatment of gasto-esophageal junction adenocarcinoma: long-term results of a single Italian centre]. AB - OBJECTIVE: The present study analysed the experience of a single Italian institution in the treatment of gastro-esophageal junction (GEJ) adenocarcinoma with the aim of assessing the long-term outcome after surgical resection. METHODS: The results of 132 patients who underwent resection with curative intent for GEJ adenocarcinoma at the First Division of General Surgery, University of Verona, from January 1988 to February 2004, were analysed statistically with special reference to Siewert type. The median follow-up period for the surviving patients was 37 months. RESULTS: Long-term survival was limited to patients who underwent RO resections (88.6%) with a 5-year survival rate of 28%. Univariate analysis showed Rp, T and pN categories to be significant prognostic factors (P<0.001), with chance of cure limited to patients with less than 6 involved lymph nodes. At multivariate analysis, R category and lymph node involvement were the most important prognostic factors while pT category lost the significance shown at univariate analysis (P=0.082). Siewert classification did not show any prognostic significance (P=0.969), but the mode of recurrence differed for the three Siewert types: in type I tumors, the majority of relapses were haematogenous (67%), while they were prevalently intra-abdominal in type III (65%) with a high rate of peritoneal carcinosis (26%). CONCLUSIONS: The long-term prognosis for GEJ cancer remains poor, independently from Siewert type, with cure limited to patients with less than 6 involved lymph nodes. PMID- 15756956 TI - [Day surgery in breast reconstructive surgery: our experience]. AB - Breast cancer is the most common tumour in Italy in the female population, counting for about 40000 new cases every year. The psychological aspects of breast mutilation and the social and economic implications are receiving increasing attention. Despite of the diffusion of screening programs to detect pre-clinical breast cancers, 30% of patients still undergo radical interventions. Therefore, many women present serious limitations of their social-life that can lead to severe depression since, in occidental countries, the biological function of the breast is less considered than its primary role of femininity and sexuality. The gold-standard is to conceal oncological radicality and aesthetic preservation. The Authors present their experience analysing the techniques employed. PMID- 15756955 TI - [Abdominal compartment syndrome: patophysiologic and clinic remarks]. AB - The abdominal compartment syndrome is a high grade abdominal hypertension with clinical evidence of multiorgan failure (MOF). It is more and more frequently observed in intensive-care units as a complication in critical patients, but especially in traumatology and surgery. The incidence is highly variable according to the different trials but the severity of scores is the common factor. All the possible mechanical, haemorrhagical, inflammatory and traumatological causes act but do not enable the stability of the abdominal content, abdominal compliance and parietal tension. The initial triad of effects consists in diaphragm elevation and visceral and vascular compression and therefore triggers a physio-pathological way that leads to a respiratory, renal and cardiovascular dysfunction and to parietal, hepatic and intestinal ischaemia and consequent bacterial translocation: sepsis and MOF. Burch's classification (1996) reports four levels of gravity from low (<15 mmHg) to severe (>35 mmHg): both of the first grades should be managed in intensive-care units with conservative pharmacological procedures, while for the two others a surgical approach of laparotomy with drainage and temporaneous closure of the abdominal wall should be considered. As mortality is still very high (29-62%), especially when multiorgan failure is already set; bladder pressure of all critical patients should be monitorized to treat immediately any potential abdominal hypertension. PMID- 15756957 TI - [Recent progress in the treatment of lower limb defects]. AB - Technological and reconstructive surgical procedures improvments are limiting demolitive interventions on the lower limb for common pathologies and traumatology. From 1999 to 2002, 32 men and 14 women, from 18 to 54 years-old, were admitted at the Operative Unit of Plastic Surgery of ASL RMH. They were grouped according to cause and site of extremity defects and different surgical techniques. Seven cross-leg flaps, 31 proximity flaps, and 18 skin grafts were performed. The results were satisfactory except in two cases. The Authors are oriented towards the cross-leg flap for foot repair. For other anatomical districts, they prefer muscular flaps covered by skin grafts. Finally, for neoplastic lesions they choose proximity flaps. PMID- 15756958 TI - [Pancreatic metastasis of renal cell carcinoma: a case report and review of the literature]. AB - The pancreas is an uncommon site of metastases from renal cell carcinoma; however in the literature late pancreatic metastases are described. In this report a 74 years-old asymptomatic man was referred for evaluation of a mass in the distal portion of the pancreas, found on CT 4 years after right nephrectomy for a renal cell carcinoma. A distal pancreatectomy and splenectomy were performed and histopathological analysis revealed to be a metastases from renal cell carcinoma. A postoperative pancreatic fistula was treated in a conservative way. The patient is alive and doing well 3 years after pancreatic surgery. This clinical report suggests that late pancreatic metastases are rare but not impossible and should be taken into consideration during a careful long-term follow-up for renal carcinoma. In addition, as it is also desumed by the Literature on survival, pancreatic metastases should be treated when possible with radical resection. PMID- 15756959 TI - [Pelvic abscesses. Treatment modalities and personal experience]. AB - The pelvic abscesses as complication of surgical operations or various pathologies is a delicate clinical situation because of the possible and unexpected evolution into a settic shock with a high risk of death. The authors report their personal experience of 16 cases undergone surgical treatment. They underline the importance of an early diagnosis, of a control of the patient general conditions with a specific antibiotic therapy, of the possibility to treat this pathology in a percutaneous way with the help of radiologic techniques or eventually by surgery. PMID- 15756960 TI - [Anaesthesiologic problems about hyperthermic intraoperative intraperitoneal chemotherapy]. AB - The literature considers hyperthermic intraoperative intraperitoneal chemotherapy a safe and effective procedure for peritoneal carcinomatosis, but a technical improvement is necessary. Regional chemotherapy anticipates the "downfall" of tumoral cells in the peritoneum. The Authors considered 5 patients--female, age 27-45 years, ASA 2--operated of peritonectomy in ovaric neoplasia with peritoneal metastasis. The hyperthermic intraoperative intraperitoneal chemotherapy has been made at the end of the surgery with a hot solution (43 degrees C): 3000 ml of dextrose 1.5% with mytomicina C 25 mg e cysplatino 75 mg/m2. We considered variation of emodinamic parametres (blood pressure, central venous pressure, stroke volume, etc.) and biochemical parametres (Na, K, CI-, CO2, etc.). These parametres have been correlated with some complications: fistula, anastomotic leakage, pancreatitis and postoperative bleeding. PMID- 15756961 TI - [Rule of early diagnosis for sigmoid volvulus. Case report]. AB - The sigmoid colon is the most frequent site for a volvulus due to anatomical and pathological reasons. When the rotation of the loop occurs slowly, the sigma can greatly blow up and stretch. We report the clinical case of an 80-year-old woman admitted to our Department for an enormous volvulus of sigmoid colon. Physical examination, abdominal X-ray and CT-scan were performed to diagnose the disease. At the surgical procedure the sigma was about 50 centimetres in length and 15 centimetres in diameter, with a gangrenous necrosis of the loop; the treatment was a sigmoidectomy by Hartmann procedure. Surgical resection is the only therapeutical option for volvulus in advanced stage due to a late diagnosis. PMID- 15756962 TI - Effect of Mycobacterium sp. strain CH1 and mycobacterium sp. strain CH2 on the degradation of four-ring creosote compounds. AB - The influence of nutrients, Mycobacterium sp. strain CH1 and Mycobacterium sp. strain CH2 on the degradation of aged creosote hydrocarbon contaminants was investigated. The Mycobacterium sp. strain CH2 showed the highest positive influence on the degradation of three- and four-ring PAH compounds. The addition of Mycobacterium sp. strain CH1 had the second highest measured positive influence on the degradation of four-ring compounds. Soluble nitrogen and phosphorus also increased the degradation of aged creosote compounds in the contaminated soil. The addition of bacteria decreased the number of measured bacterial species, indicating competition for limited nutrients in the soil. PMID- 15756963 TI - Mass transfer of polyaromatic hydrocarbons (PAHs) in a two-liquid-phase system. AB - Studies were conducted to observe the solubility of phenanthrene, anthracene, and pyrene in four water-immiscible liquids (WILs) and the mass transfer of PAH in a two-liquid-phase (TLP) system. The order of solubility in WILs for three different PAHs was phenanthrene > pyrene >> anthracene. Among the WILs tested in a TLP system, paraffine oil did not affect the mass transfer of three PAHs, while silicone oil reduced the mass transfer of anthracene significantly because paraffine oil had a higher solubilizing capacity for anthracene than silicone oil. When initial PAHs concentration increased up to 2500 mg PAHs/kg soil, the mass transfer of phenanthrene and pyrene were hardly influenced, while that of anthracene was reduced to 40%. As the amount of light paraffine oil decreased, the mass transfer of anthracene decreased significantly compared to that of phenanthrene and pyrene. Based on the present results, it was confirmed that the mass transfer of PAHs from soil to WIL was governed mainly by the solubilizing capacity of WIL for PAHs. This is the first report on the mass transfer of PAHs from soil to WIL, in which the mechanism of TLP process can be understood more clearly. PMID- 15756964 TI - Effects of alum coagulation on speciation and distribution of trihalomethanes (THMs) and haloacetic acids (HAAs). AB - The impacts of alum coagulation on the distribution of disinfection by-products (DBPs), trihalomethanes (THMs), and haloacetic acids (HAAs) were evaluated under controlled chlorination conditions using four surface waters. Among the nine HAAs found in waters, dihaloacetic acids (X2AAs) have been found to be the dominant species in all of the raw and alum treated waters. Alum coagulation tends to remove more monohaloacetic acids (XAAs) and trihaloacetic acids (X3AAs) precursors than that of dihaloacetic acids (X2AAs). Alum coagulation treated water had a lower HAA9/TTHM ratio compared with that of the raw water. The increase of THM bromine incorporation factors (BIFalpha) value of alum treated water was statistically significant in comparison with the raw water. On average, BIFalpha increased by 54% after the alum coagulation process in these four different waters. This indicated that THM speciations shifted in favor of the more brominated compounds. However, alum coagulation treatment process had less effect on HAA bromi ne incorporation factors (BIFbeta)than it did on BIFalpha. Bromine incorporation factor (BIF) values decreased with time in the THM and HAA formation processes, especially within the first 10 h of the reaction time. This suggested that brominated THMs or HAAs formed faster than the chlorinated species in the initial period. PMID- 15756965 TI - Evidence for tin inhibition of enhanced biological phosphorus removal at a municipal wastewater treatment plant. AB - Concentrations of 34 metals were determined in the concentrated waste activated sludge from a dissolved air flotation unit at an advanced municipal wastewater treatment plant (WWTP) with biological nutrient removal. Reduction in enhanced biological phosphorus removal (EBPR) efficiency was observed at total tin concentrations greater than 4 microg L(-1) in the solids fraction of the mixed liquor suspended solids. No influence on carbon or nitrogen removal efficiency was found by elevated tin concentrations. Other process control variables and metal concentrations were not correlated with reduced EBPR efficiency on dates with elevated tin levels. The known high contributions of organotin species toward total tin in activated sludges from other municipal WWTPs, and the high toxicity of these compounds, suggests elevated organotin levels may be responsible for the observed reduction in EBPR efficiency. PMID- 15756966 TI - Desulfurization of Mexican heavy oil by sulfate-reducing bacteria. AB - Twenty-five mixed cultures of sulfate reducing bacteria (SRB) were isolated from sediment and anaerobic digestors samples, collected at southeast Gulf of Mexico, Pacific Ocean, and wastewater treatment plant, Mexico. The isolated SRB mixed cultures were tested for desulfurization of Mexican heavy oil. Desulfurization activity of SRB was not affected by high level of vanadium in heavy oil. Sediment samples gave better sulfur removal performance than anaerobic digestors samples. The difference in removal efficiency of the two samples is possibly related to the different quantity of SRB strains causing degradation of organic sulfur in heavy oil. PMID- 15756967 TI - Monitoring of air pollution in the atmosphere around Oman Liquid Natural Gas (OLNG) plant. AB - This study was basically designed to assess the potential environmental air quality impacts arising from the existing two operational trains at the Oman Liquid Natural Gas (OLNG) plant. The results of the paper contain a baseline survey of the existing environment. The pollutants studied included methane (CH4), non-methane hydrocarbons (NMHC), carbon monoxide (CO), nitrogen oxides (NOx), and suspended particulate matters (dust PM 10). Meteorological parameters monitored simultaneously include wind speed and direction, air temperature, and relative humidity. The air quality data were used to determine the diurnal and monthly variations in the pollutants. Description levels of the pollutants with respect to meteorological data were also used in analysis. Moreover, a statistical analysis of the collected data was presented. Generally, the results indicated that the mean concentrations of pollutants were low to cause any significant impact in air quality. The area had no problem in meeting the air quality standards for CO and NO2. It was also found that there was a random relationship between CO and NMHC, and between NO and NOx (no apparent correlation). The diurnal peaks of NOx, NO2, THC, and NMHC over a 24-h period were observed at around 9:00-10:00 AM (morning peak). For NO, NO2, and NOx, another peak was seen at around 5:00 PM (evening peak). Furthermore, the measured concentrations for NO2, NOx, and CO were found higher in winter than in summer. The study would help to gain a better understanding of local background levels of air pollutants at the area prior to the construction of new industrial projects, and to prepare action plans for controlling pollution in the area. PMID- 15756968 TI - Reconstruction of air contaminant concentration distribution in a two-dimensional plane by computed tomography and remote sensing FTIR spectroscopy. AB - This research combined open path FTIR (OP-FTIR) technique and computed tomography (CT) to reconstruct air contaminant concentration distribution in a two dimensional plane. Remote sensing FTIR instrument was used to scan radial beam geometry and obtain path integrated concentration (PIC) data of acetone gas in the measuring plane. Smooth basis function minimization (SBFM) algorithm was adopted to reconstruct gaseous concentration distribution. For the purpose of finding out the preferable number of Gaussians used in SBFM algorithm, single Gaussian, double-Gaussian, and three-Gaussian models were used respectively. Experimental results showed that the reconstruction result of acetone concentration distribution by SBFM algorithm with double-Gaussian model agreed with real distribution more qualitatively and quantitatively than single-Gaussian and three-Gaussian. Also, it has been proved that simulated annealing algorithm used in the optimization process of SBFM reconstruction was feasible and effective. Although computed tomography and remote sensing FTIR technique (CT-RS FTIR) is still at the laboratory study stage, with further improvement of SBFM algorithm and beam geometry, it promises to be used in air pollution monitoring widely. PMID- 15756969 TI - Sorption of 2,4,6-trinitrotoluene to natural soils before and after hydrogen peroxide application. AB - Laboratory batch sorption experiments were conducted to investigate the impact of hydrogen peroxide (H2O2) pre-application on post-sorptive behavior of 2,4,6 trinitrotoluene (TNT) in different natural soils (average soil, high Fe soil, and high pH soil). After H2O2 application, the values of Freundlich coefficient Kf were increased by approximately 160% for the average and high pH soils and by approximately 120% for the high Fe soil, showing that the soils became more favorable for TNT sorption after H202 application. Nonlinearity in terms of the Freundlich exponent n was increased by approximately 40% for the average and high pH soils and by approximately 30% for the high Fe soil, showing greater sorption affinity of TNT for the oxidized soils at lower TNT concentrations and also implying greater TNT availability for transport at high concentrations. The increase in sorption extent for the H2O2-oxidized soils was presumably attributed to the oxygen-induced enhancement in the sorption capacity of the soils and the more dominant contribution of clay minerals to sorption. Therefore, enhanced sorption following H2O2 application may inhibit the subsequent formation of a TNT plume after either source zone remediation or plume remediation using H2O2 such as Fenton oxidation. PMID- 15756970 TI - Distribution of cadmium and lead in liver and kidney of some wild animals in Slovakia. AB - The content of cadmium and lead, as risk factors of environment, in liver and kidneys of wild animals as brown hare (Lepus europaeus), yellow-necked mouse (Apodemus flavicollis), wood mouse (Cleithrionomys glareolus), and red deer (Cervus elaphus) were studied. Samples were analyzed by the atomic absorption spectrophotometry method (AAS). The highest levels of cadmium were found in kidneys (0.213-2.387 mg/kg) of all animal species. The concentration of cadmium in liver was 0.032-0.258 mg/kg. The analysis of lead showed that the concentration of this element was higher in kidneys of yellow-necked mouse and wood mouse (0.503-0.780 mg/kg) than in liver (0.177-0.268 mg/kg). In brown hare and red deer a higher accumulation of lead in liver (0.221-1.904 mg/kg) in comparison with kidneys (0.115-0.561 mg/kg) is reported. PMID- 15756971 TI - Extraction of selected heavy metals using modified clays. AB - In the present study, attapulgite, kaolinite, and montmorillonite KSF were modified using azeotropic distillation to condense 2-mercaptoethanol with the clay material. The resulting product was used as a coordinating agent to remove selected metal ions, e.g., copper(II), cadmium(II), silver(I), nickel(II), and lead(II) ions from standard aqueous solutions. Batch systems were used, and samples were shaken for two hours, and following filtration, metal content of the filtrate was measured by atomic absorption spectrometry. Without adjusting the pH, better than 90% of the metal ions could be removed. PMID- 15756972 TI - Rapid determination of total Kjeldahl nitrogen using microwave digestion. AB - A closed-vessel microwave digestion process for the determination of total Kjeldahl nitrogen (TKN) has been developed for sewage and wastewater. TKN values obtained from the microwave digestion method were in excellent agreement with those of the thermal digestion method. The accuracy of both analytical methods is comparable. In comparison to the conventional thermal digestion, the microwave method shortened the time required for complete digestion from 4 h to 25 min, and also decreased the maximum digestion temperature from 380 degrees C to 200 degrees C. This developed method may contribute to a significant reduction in sample digestion time, resulting in an increase in analytical throughput. The microwave digestion method developed in this study could be a rapid and efficient means for TKN determination for sewage wastewater and sludge. PMID- 15756973 TI - Study on the recovery of phosphorus from waste-activated sludge incinerator ash. AB - Several batch studies that were made up of the acid extraction and the solvent extraction were performed to recover phosphorus from the waste-activated sludge (WAS) incinerator ash. In the acid extraction, the extraction efficiency of phosphorus relied on the acid type, liquid(acid)-to-solid (L(acid)S) ratio, and acid concentration. Phosphorus in the WAS incinerator ash was completely extracted by 1 M HCl at the L(acid)S ratio of 6.4:1. Subsequently, the solvent extraction was conducted to separate and concentrate phosphorus further from the acid extract. The efficiency of solvent extraction was affected mainly by the solvent type, liquid (solvent)-to-liquid (the acid extract) (L(solv)L(acid ext)) ratio, and hydrogen ion concentration. Under the appropriate condition, 76% of phosphorus in the acid extract was extracted to 1-butanol phase, which corresponded to 80.1% as the mass fraction of phosphorus to total elements. Prior to the solvent extraction, the addition of bis (2-ethylhexyl) phosphoric acid (D2EHPA), which was available for removing aluminum from the acid extract, led to an additional increase in the term of the mass fraction of phosphorus to total elements. Overall results indicated that phosphorus in the WAS incinerator ash could be efficiently recovered and be a potential renewable resource. PMID- 15756974 TI - Inhibition of volatile fatty acid production in granular sludge from a UASB reactor. AB - Inhibition of volatile fatty acids (VFA), namely acetate, butyrate, and propionate, on the activity of acetoclastic methanogens within a full-scale upflow anaerobic sludge blanket (UASB) reactor was investigated using specific methanogenic activity (SMA) test. SMA tests were carried out at acetate concentrations in a range of 1000-25,000 mg l(-1), butyrate concentrations in a range of 3000-25,000 mg l(-1) and propionate concentrations between 500-10,000 mg l(-1). Maximum potential methane production (PMP) rates were obtained as 389 ml CH4 gTVS(-1) x d(-1) at 3000 mg l(-1) acetate concentration, 432 ml CH4 gTVS(-1) x d(-1) at butyrate concentration of 5000 mg l(-1), and 162 mlCH4 gTVS(1) x d(-1) at 1000 mg l(-1) propionate concentration. App. 50% and 100% inhibition occurred at acetate concentrations of 13,000 mg l(-1) and 25,000 mg l(-1), butyrate concentrations of 15,000 mg l(-1) and 25,000 mg l(-1), and propionate concentrations of 3500 mg l-1) and 5000 mg l(-1), respectively. PMID- 15756975 TI - Studies on the reuse of waste printed circuit board as an additive for cement mortar. AB - The recent development in electronic industries has generated a drastic increase in production of printed circuit boards (PCB). Accordingly, the amount of waste PCB from electronic productions and waste electronics and its environmental impact such as soil and groundwater contamination have become a great concern. This study aims to propose a method for reuse of waste PCB as an additive for cement mortar. Although the expansibility of waste PCB powder finer than 0.08 mm in water was observed to be greater than 2.0%, the maximum expansion rates in water for 0.08 to approximately 0.15 and 0.15 to approximately 0.30 mm sized PCB powders were less than 2.0%, which satisfied the necessary condition as an alternative additive for cement mortar in place of sand. The difference in the compressive strength of standard mortar and waste PCB added mortar was observed to be less than 10% and their difference was expected to be smaller after prolonged aging. The durability of waste PCB added cement mortar was also examined through dry/wet conditioning cyclic tests and acidic/alkaline conditioning tests. From the tests, both weight and compressive strength of cement mortar were observed to be recovered with aging. The leaching test for heavy metals from waste PCB added mortar showed that no heavy metal ions such as copper, lead, or cadmium were detected in the leachate, which resulted from fixation effect of the cement hydrates. PMID- 15756976 TI - Solubility of sulfur dioxide in chlorine-treated wastewater. AB - The solubility of sulfur dioxide in chlorine-treated wastewater was investigated. An experimental study was carried out to measure the solubility of sulfur dioxide in chlorine-treated wastewater over the temperature range of 10-40 degrees C. The highest solubility of sulfur dioxide was found to be equal to 1.098 x 10(-3) mole/L at 10 degrees C, whereas the lowest value of 6.17 x 10(-4) mole/L was obtained at 40 degrees C. The solubility of sulfur dioxide was correlated as a second-order dependence on temperature. The measured solubility data of sulfur dioxide in chlorine-treated wastewater were found in good agreement with that obtained using the solubility correlation. PMID- 15756977 TI - Is biodegradation of bitumen a source of recalcitrant naphthenic acid mixtures in oil sands tailing pond waters? AB - Carboxylic acids are transient metabolites during the mineralization of petroleum hydrocarbons. Crude oils, however, vary in their proportion of the hydrocarbon components. Depending on structure, some carboxylic acid metabolites resist further biodegradation and persist in aquatic systems. During the extraction of oil sands bitumen, recalcitrant carboxylic acid mixtures, collectively referred to as naphthenic acids (NAs), are released into the wastewaters. These waters also contain unrecovered bitumen from the oil sands. The unextracted bitumen is often overlooked as a possible source of the petroleum acids. The present article discusses the literature data on the biotransformation of hydrocarbons in bitumen from oil sands to the corresponding petroleum carboxylic acids. Some insight is given on the mechanism of the biodegradation process. The susceptibility to biodegradation is affected by differences in alicyclic carboxylic acids such as carbon chain length, chain branching, and the oddness or evenness of carbon chain containing the carboxylic group, positions where alkyl groups are substituted on the cyclic ring, geometrical isomerism, and number of cyclic rings. PMID- 15756978 TI - In situ bioremediation of naphthenic acids contaminated tailing pond waters in the athabasca oil sands region--demonstrated field studies and plausible options: a review. AB - Currently, there are three industrial plants that recover oil from the lower Athabasca oil sands area, and there are plans in the future for several additional mines. The extraction procedures produce large volumes of slurry wastes contaminated with naphthenic acids (NAs). Because of a "zero discharge" policy the oil sands companies do not release any extraction wastes from their leases. The process-affected waters and fluid tailings contaminated with NAs are contained on-site primarily in large settling ponds. These fluid wastes from the tailing ponds can be acutely and chronically toxic to aquatic organisms, and NAs have been associated with this toxicity. The huge tailings containment area must ultimately be reclaimed, and this is of major concern to the oil sands industry. Some reclamation options have been investigated by both pioneering industries (Syncrude Energy Inc. and Suncor Inc.) with mixed results. The bioremediation techniques have limited success to date in biodegrading NAs to levels below 19 mg/L. Some tailing pond waters have been stored for more than 10 years, and it appears that the remaining high molecular weight NAs are refractory to the natural biodegradation process in the ponds. Some plausible options to further degrade the NAs in the tailings pond water include: bioaugmentation with bacteria selected to degrade the more refractory classes of NAs; the use of attachment materials such as clays to concentrate both the NA and the NA-degrading bacteria in their surfaces and/or pores; synergistic association between algae and bacteria consortia to promote efficient aerobic degradation; and biostimulation with nutrients to promote the growth and activity of the microorganisms. PMID- 15756979 TI - Gerontological nursing and the aging work force. PMID- 15756980 TI - Evidence-based administrative guideline: quality improvement in nursing homes. AB - Those staff members involved in a quality improvement program for a nursing home play a significant role in ensuring quality care for older adults. Use of the evidence-based administrative guideline "Quality Improvement in Nursing Homes" (Dyck, 2003) can provide a template for the implementation of a research-based quality program. PMID- 15756981 TI - The meaning of mealtimes: connection to the social world of the nursing home. AB - Food that reflects our family backgrounds is a source of comfort that can play an important part in recovery from illness or adaptation to the nursing home, especially for older individuals. However, no studies could be found that explored residents' perspectives on how their food and food service preferences are, or are not, met in nursing homes. This exploratory qualitative study examined dietary preferences acquired during the course of a lifetime, and the meaning of mealtimes to 20 nursing home residents, and attempted to connect that meaning with their social world. Exploring the meaning of food and food service to nursing home residents could furnish insights for improving nutritional status, adaptation to the nursing home, and quality of life through promotion of individualized nutritional care. PMID- 15756982 TI - Nurse staffing in nursing homes in the United States. AB - This study described the nurse staffing standards in nursing homes in all states and the District of Columbia in 2000 to 2001. An Internet survey of state statutes and regulations and a structured telephone survey of state officials were used to collect data for this descriptive study. Almost half of the states had higher licensed nurse (registered nurse and licensed vocational nurse) requirements than the federal government and 33 states had established minimum state requirements for direct care workers (including nursing assistants). Thirteen states had increased their state standards in the 1999 to 2001 period. Federal standards for nursing homes lag behind those in many states. PMID- 15756983 TI - Fictive kin: friends as family supporting older adults as they age. AB - Assessing social support or social capital is receiving renewed attention in health care. A search of the nursing literature revealed limited attention to a potential resource in the care of older adults--fictive kin. Transdisciplinary searches indicated that "fictive kin" or "chosen kin" are increasingly important as family size shrinks and older adults live longer lives. Fictive kin are common across cultural groups and some have designated special names for these quasi family members. The purpose of this article is to advance the concept of fictive kin as a resource for older adults and to provide strategies to promote engagement of fictive kin in health care planning by nurses. PMID- 15756984 TI - Ethical issues related to health care: the older adult's perspective. AB - Although ethical issues in health care receive much publicity, attention is rarely given to the non-dramatic, everyday ethics of health care. This American Nurses' Foundation (ANF) funded pilot study was significant because it was the first to investigate everyday ethical issues from the perspective of older adult health care consumers. The overall goals of this descriptive study were to pilot interview questions and study design, as well as collect preliminary data about the ethical issues older adults in two settings (a residential facility and a non residential community center) encounter related to health care. A sample of 10 participants answered several open-ended questions in an audiotaped interview; the tape was subsequently transcribed verbatim and analyzed using content analysis methods. Although two open-ended questions were effective in eliciting ethically related health care issues, the addition of specific examples to those questions was suggested. Health care related ethical issues identified by the older adults were categorized as Attentiveness, Respect, Care, Finances, and Health Care Systems. Although the residential participants did not experience financial issues as did the community participants, the highest priority issues for both residential and community participants were in the Attentiveness and Respect categories. The clinical implication of these findings is that nurses best address older adult health care consumers' most important ethical issues by providing attentive, respectful care in all they do. PMID- 15756985 TI - The context for nursing home resident care: the role of leaders in developing strategies. AB - Data obtained from three Midwestern nursing homes were used to identify and describe contextual factors that influenced resident care. A qualitative, emergent, case study design guided the study. In addition to 17 residents and 16 family members or friends of the respective residents, participants included 66 staff members and 9 managers involved in the planning and delivery of care to the participating residents. Data were collected via observation, semi-structured interview, and resident record audit. Contextual factors seemed to integrate or fragment care planning and delivery. Important integrators were shared values, the role of the Minimum Data Set coordinator, the role of other leaders, and family influence. Two fragmenting factors, competing demands for the staff members' time and a task orientation to care, appeared to be ubiquitous. External accountability also fragmented care to a discernible degree. PMID- 15756986 TI - Risk factor for accidental injuries within senior citizens' homes: analysis of the Canadian Survey on Ageing and Independence. AB - Using data from the Survey on Ageing and Independence (SAI), risk factors for unintentional injuries occurring within the homes of individuals older than 65 are identified. For the SAI, conducted by Statistics Canada in 1991, data were collected on a representative sample of approximately 20,000 individuals between ages 45 and 102. For each household contacted, one individual older than 45 was interviewed via the telephone. For the present analysis, only individuals older than 65 (n = 10,059) were used. Approximately 5% of senior citizens experienced an injury that limited their activity for at least 1 day. Using logistic regression, the following risk factors for injury were identified: education, alcohol consumption, smoking, rest and sleep patterns, support, and interactions between age and gender, activity limitations and age, and home maintenance and gender. The present findings are important to the body of research concerning injuries among older adults. The results expand current univariate analysis of data identifying risk factors for injuries within the literature and provide comprehensive information pertaining to risk factors for accidental injuries at the multivariate level. Identification of risk factors provides health care professionals, particularly front line nurses, with insight into factors that, if modified, have the potential to decrease accidental injuries and improve or maintain quality of life. PMID- 15756987 TI - Old Willie. PMID- 15756988 TI - Clinical information systems. Putting patients first. PMID- 15756989 TI - Clinical information systems HotList. PMID- 15756990 TI - Internet applications/ASPS. Nothing but 'net. PMID- 15756991 TI - Decision support. More than a pocketful of knowledge. PMID- 15756993 TI - Health plans & technology. Processing for profitability. PMID- 15756992 TI - Physician practice management. The path to the future. PMID- 15756994 TI - Speech recognition/transcription. Is speech recognition the Holy Grail? PMID- 15756995 TI - Business process management. Waves of efficiency. PMID- 15756996 TI - Supply chain management. Eliminating sticker shock. PMID- 15756997 TI - Clinical information systems. Against the grain. PMID- 15756998 TI - The future of CIOs. PMID- 15756999 TI - The CMS update cycle: short, short-sighted and short-changing the industry. PMID- 15757000 TI - Time-series analysis of air pollution and mortality: a statistical review. AB - The Walter A Rosenblith New Investigator Award provided funding to explore new statistical approaches for air pollution research. This report reviews and summarizes the methodologic and substantive contributions to time-series analyses of air pollution and mortality that this award made possible. The review is organized according to the following general topics: (1) semiparametric methods for time-series analyses of air pollution and mortality; (2) explorations into the sensitivity of generalized additive models (GAMs*) applied to time-series data; (3) combining information in multisite time-series studies; (4) effects of misclassification of exposure; (5) mortality displacement; (6) shape of the concentration-response curve; and (7) ongoing projects and future directions. Appendix A includes abstracts of papers published as reports and in peer-reviewed journals. PMID- 15757001 TI - Comparing personality characteristics of juvenile sex offenders and non-sex offending delinquent peers: a preliminary investigation. AB - There exists a paucity of research on psychopathology among Juvenile Sex Offenders (JSOs) as measured by standardized instruments, particularly as compared to nonclinical and other clinical populations. The current investigation involved a comparison of scores on the Minnesota Multiphasic Personality Inventory-Adolescent Version (MMPI-A) between a group of JSOs and non-sex offending delinquent peers. Thirty-six youth from a staff-secure residential facilitate participated, 18 JSOs and 18 non-sex offending delinquent peers. Results revealed that the mean score on clinical Scale 4 (Psychopathic Deviant) was in the clinically significant range for non-sex offending peers and not for JSOs, although the difference between the mean scores was not statistically significant. Chi-square analysis indicated that a significantly greater number of non-sex offending delinquents had scores in the clinical range on this scale as compared to JSOs. Results are discussed in the context of current research on psychopathology among JSOs. PMID- 15757002 TI - Factor structure and validation of the juvenile sex offender assessment protocol (J-SOAP). AB - The Juvenile Sex Offender Assessment Protocol (J-SOAP) was first developed in 1994 in response to the need for a structured method of assessing risk of recidivism among juvenile sexual offenders (R. A. Prentky, B. Harris, K. Frizzell, & S. Righthand, 2000). Over the ensuing years the scale has been subjected to a series of studies that have sought to examine its psychometric properties and its validity. The current paper reports on results from three of these studies, one of which looks at the factor structure of the items. The other two studies examine concurrent validity by looking at J-SOAP Scales 1 and 2 in relation to the Level of Service Inventory-Youth Version with: Youth Level of Service/Case Management Inventory (YLSICMI) (R. D. Hoge & D. A. Andrews, 1996) and other selected variables and discriminant validity by looking at the ability of J-SOAP scales to differentiate between juvenile sex offenders in the community and juvenile sex offenders in residential placement. The revision of the scale is discussed based on the research that has developed the scale and the research that has employed it. PMID- 15757003 TI - Recovering memories of the offense in "amnesic" sexual offenders. AB - This paper describes a technique designed to assist sexual offenders to recover memories of their offense. We have consistently observed that some sexual offenders present as having no recall of their offense although they are able to remember other events of the day of the offense. This failure to recall offense details prevents the offenders from making an appropriate disclosure which, in turn, blocks attempts to identify their offense pathways and develop relapse prevention plans. The memory recovery technique we describe is based on experimental literature on memory and we outline its use with 22 clients, 20 of whom showed satisfactory recovery of their memories. PMID- 15757004 TI - Biological fathers and stepfathers who molest their daughters: psychological, phallometric, and criminal features. AB - A sample (N=143) of men who sexually abused their biological daughters or their step/adopted daughters were examined on a comprehensive array of demographic and historical information, offense characteristics, psychological and phallometric measures, and recidivism. Biological fathers were significantly less sexually aroused by children than were the stepfathers. On all the remaining variables, however, no statistically significant differences were found. Overall, biological and stepfathers in the present study appear to be much more similar than different from one another. PMID- 15757005 TI - Constructions of therapist-client sex: a comparative analysis of retrospective victim reports. AB - Former patients' (n=24) accounts of their experience during a sexual liaison with their psychotherapist (Therapist-Client Sex, TCS) could be classified as either romantic (TCS-Romance) or as an abusive encounter (TCS-Abuse). During TCS, individuals in the TCS-Romance group reported having experienced overall better emotional states and more favorable perceptions of both the perpetrating therapists and the treatments they provided. Pre-TCS assessments showed that initially TCS-Romance subjects may have had a relatively higher regard for their perpetrators and the quality of their treatments than TCS-Abuse subjects, but these between-group differences disappeared when the same variables were assessed for two post-TCS periods. These periods were marked in both groups by deteriorated indices of psychological well-being. These findings suggest that a romantic narrative of TCS could have a shielding, albeit temporary, impact on the subjective experience of what is otherwise considered an abusive relationship. PMID- 15757006 TI - Developing empathy in sexual offenders: the value of offence re-enactments. AB - This paper describes an evaluation of different uses of role-play to enhance victim-specific empathy in sexual offenders. Thirty-three men participated in a treatment program involving offence re-enactment as described by Pithers (1994) and Mann, Daniels, and Marshall (2002). A matched group of 33 men participated in a treatment program that was identical in all respects except that they did not complete offence re-enactments. Instead, they completed extra role-plays designed to enhance empathy for the short and long-term consequences for their victim(s). Results indicated that completing an offence re-enactment led to slightly better ability to identify some types of negative consequences for abuse victims, and identify cognitive distortions about their offending and women per se. Rapists in particular seemed more likely to benefit from offence re-enactment. The non reenactment group showed better understanding of lifestyle disruption effects for sexual abuse victims. The differences between the groups were not very marked, and the study only involved measures of cognitive empathy. Given the concerns about offence re-enactment expressed by Pithers (1997), this procedure should be used with caution and future investigations should test specifically for possible signs of damage caused by the procedure. PMID- 15757008 TI - Arne Tiselius--father of electrophoresis. PMID- 15757007 TI - Effects of a relapse prevention program on sexual recidivism: final results from California's sex offender treatment and evaluation project (SOTEP). AB - Final results from a longitudinal investigation of the effectiveness of cognitive behavioral treatment with sexual offenders are presented. The study was a randomized clinical trial that compared the reoffense rates of offenders treated in an inpatient relapse prevention (RP) program with the rates of offenders in two (untreated) prison control groups. No significant differences were found among the three groups in their rates of sexual or violent reoffending over an 8 year follow-up period. This null result was found for both rapists and child molesters, and was confirmed in analyses using time to reoffense as the outcome and those controlling for static risk differences across the groups. Closer examination of the RP group's performance revealed that individuals who met the program's treatment goals had lower reoffense rates than those who did not. Although our results do not generally support the efficacy of the RP model, they do suggest a number of ways in which this kind of treatment program can be improved. This study also emphasizes the importance of including appropriate control groups in treatment outcome research. Additional controlled investigations are needed to address the many questions that remain about when and how treatment works for sexual offenders. PMID- 15757009 TI - Dietary supplements and stroke. PMID- 15757010 TI - Stress imaging in chest pain units: is less more? PMID- 15757011 TI - The problems of organ donor shortage. PMID- 15757012 TI - Effect of exercise treadmill testing and stress imaging on the triage of patients with chest pain: CHEER substudy. AB - OBJECTIVE: To determine whether stress imaging for patients who are unsuitable for exercise treadmill testing (ETT) as part of a chest pain unit (CPU) triage strategy resulted in incremental benefit in clinical outcomes and relative costs compared with patients randomized to routine hospital admission. PATIENTS AND METHODS: Clinical outcomes and medical resource utilization were examined at the Mayo Clinic in Rochester, Minn, for 212 intermediate-risk patients with unstable angina randomized to a CPU and compared with 212 patients randomized to routine admission from November 21, 1995, to March 18, 1997. Patients in stable condition in the CPU underwent ETT; if patients were unsuitable for ETT, stress imaging was performed. Costs for CPU evaluation and outcomes were assessed during a 6-month follow-up. RESULTS: During the observation period, 60 patients (28%) were admitted to the hospital. Of the 152 remaining patients, 125 (82%) underwent ETT (91 had normal results), and 27 (18%) underwent stress imaging (3 had normal results). Patients with normal ETT or stress imaging results had no primary events at 6-month follow-up. Patients admitted to the hospital who underwent stress imaging had an insignificantly higher 6-month event rate compared with patients who underwent ETT (16.7% vs 8.1%; P=.38). The standardized resource based relative-value units (RBRVUs) for patients who underwent ETT and stress imaging during follow-up were 19.4 and 56.4 RBRVUs, respectively, compared with 51.4 (ETT) and 52.1 (stress imaging) RBRVUs for similar numbers of patients randomized to routine admission. CONCLUSIONS: Exercise treadmill testing safely stratified most intermediate-risk patients with unstable angina and was less costly than routine admission. Patients not suitable for ETT are likely to have abnormal stress imaging results. They represent a higher-risk cohort that could be routinely admitted to the hospital without reducing the effectiveness of the CPU strategy. PMID- 15757013 TI - Evaluation of orthostatic hypotension: relationship of a new self-report instrument to laboratory-based measures. AB - OBJECTIVE: To compare measured autonomic deficits (composite autonomic severity score [CASS]) with a brief self-report scale we developed to measure severity of symptoms of orthostatic hypotension. PATIENTS AND METHODS: Patients were recruited in 2 phases: from August to October 2002 and in April 2004. All patients underwent full evaluation in the autonomic laboratory, from which a CASS of autonomic deficits was derived. Patients also completed the 5-item self-report Orthostatic Grading Scale, which inquires about symptoms of orthostatic intolerance due to orthostatic hypotension (eg, severity, frequency, and interference with daily activities). RESULTS: Of 145 patients, 97 (67%) had orthostatic hypotension. The 5-item scale demonstrated strong internal consistency (coefficient alpha=.91). Patients with orthostatic hypotension had significantly higher scores on each questionnaire item and CASS subscores than those without orthostatic hypotension. The scale items correlated significantly with each of the CASS subscores, maximally with the CASS adrenergic subscore. CONCLUSIONS: Orthostatic hypotension is not the only cause of reduced orthostatic tolerance, and some patients may have orthostatic hypotension but be asymptomatic. Results of this study indicate that this 5-item questionnaire is a reliable and valid measure of the severity of symptoms of orthostatic hypotension and that it can supplement laboratory-based measures to provide a rapid, more complete clinical assessment. This questionnaire would also be useful as a brief screening device for orthostatic intolerance to aid physicians in identifying patients who may have orthostatic hypotension. PMID- 15757014 TI - Prevalence of hostility in young coronary artery disease patients and effects of cardiac rehabilitation and exercise training. AB - OBJECTIVE: To determine the prevalence of hostility symptoms in young patients with coronary artery disease (CAD), the associated risk factor profile in these patients, and the effects of a formal phase 2 cardiac rehabilitation and exercise training program. PATIENTS AND METHODS: Our study included consecutive CAD patients referred for cardiac rehabilitation from May 1999 through December 2000. At baseline and after rehabilitation, behavioral factors and quality of life were assessed by validated questionnaires, and standard CAD risk factors were measured, including exercise capacity. We specifically evaluated detailed data for young patients. RESULTS: A total of 500 patients were included in the study. Hostility scores were 2.5 times higher (P<.001) in the 81 young patients (< 50 years; mean +/- SD age, 45 +/- 5 years) than in the 268 elderly patients (> or = 65 years; mean +/- SD age, 70 +/- 4 years), and the prevalence of hostility symptoms was 3.5 times higher in young patients (28% vs 8%; P<.001). Young patients with hostility symptoms also had more adverse CAD risk profiles, including higher total cholesterol levels, triglyceride levels, total cholesterol/high-density lipoprotein cholesterol ratios, fasting glucose levels, and glycosylated hemoglobin levels and lower quality-of-life scores compared with young patients with low hostility scores. After cardiac rehabilitation, young patients with hostility symptoms had marked improvements in CAD risk factors, behavioral characteristics (including hostility), and quality of life, and a nearly 50% (P=.005) reduction in the prevalence of hostility symptoms occurred. CONCLUSIONS: Young CAD patients have a high prevalence of hostility symptoms and adverse CAD risk profiles. Reducing hostility symptoms and other parameters of psychological distress in young CAD patients should be emphasized, and the potential benefits of cardiac rehabilitation programs in the secondary prevention of CAD should be highlighted. PMID- 15757015 TI - Effect of alendronate on vertebral fracture risk in women with bone mineral density T scores of-1.6 to -2.5 at the femoral neck: the Fracture Intervention Trial. AB - OBJECTIVES: To determine the efficacy of alendronate treatment on risk of vertebral fracture in a subgroup of women from the Fracture Intervention Trial who had bone mineral density T scores between -1.6 and -2.5 at the femoral neck and to describe how soon after initiation of therapy alendronate becomes effective and whether it is consistent in women with and without existing radiographic vertebral fracture. PATIENTS AND METHODS: From May 1992 to March 1997, postmenopausal women aged 55 to 80 years were randomized to receive alendronate at 5 mg/d for 2 years and 10 mg/d thereafter or placebo for up to 4.5 years (mean, 3.8 years) in a controlled, double-blind, multicenter study. RESULTS: A total of 3737 postmenopausal women were included in the study, 1878 in the alendronate group and 1859 in the placebo group. Risk of vertebral fracture was significantly reduced by alendronate compared with placebo for clinical (relative risk [RR], 0.40; 95% confidence interval [CI], 0.19-0.76; P=.005) and radiographic (RR, 0.57; 95% CI, 0.41-0.81; P=-.002) fracture. The reductions in vertebral fracture risk were consistent in women with and without an existing radiographic vertebral fracture for clinical (RR, 0.34; 95% CI, 0.12-0.84; and RR, 0.46; 95% CI, 0.16-1.17; respectively) and radiographic (RR, 0.53; 95% CI, 0.34-0.82; and RR, 0.64; 95% CI, 0.38-1.10; respectively) fractures. In both groups, the effect of alendronate on clinical vertebral fracture was noted soon after therapy was initiated. The absolute risk of vertebral fracture was low in women without a baseline radiographic fracture. CONCLUSIONS: In women with low bone mass who do not meet the bone mineral density criterion for osteoporosis, alendronate is effective in reducing the risk of vertebral fractures. The absolute benefit of this therapy in women with a T score between -1.6 and -2.5 is greater in women with an existing vertebral fracture and/or with other risk factors. The effect of alendronate occurs early. PMID- 15757016 TI - Postoperative stupor and coma. AB - OBJECTIVES: To identify predictive factors for postoperative coma or stupor and to examine the value of neuroimaging techniques in elucidating structural brain damage. PATIENTS AND METHODS: We performed a case-control study of surgical patients admitted to a Mayo Clinic-affiliated hospital. We studied preoperative comorbidity, intraoperative hypotension, and postoperative data in patients with postoperative stupor or coma and compared the characteristics with control patients (surgical intensive care unit patients with neurologic consultations for other reasons). RESULTS: A total of 35 patients with stupor or coma after surgery and 31 control patients participated in this study. Comatose patients were older (P=.004) and had significantly more presurgical comorbidity (P<.001), cardiovascular surgical procedures (P<.001), and intraoperative hypotension (P=.03). Adjusted for age and comorbidity, intraoperative hypotension remained statistically significant but not after adjusting for cardiovascular surgery. Of the 34 computed tomograms obtained, 41% showed abnormal results; of the 12 magnetic resonance images obtained, 58% showed abnormal results. Both showed primarily infarctions. In 4 patients with normal computed tomographic results, magnetic resonance imaging showed multiple territorial infarctions. CONCLUSION: Prior comorbidity, older age, intraoperative hypotension, and cardiovascular surgery may predispose patients to postoperative coma. Widespread structural ischemic brain damage was often documented by neuroimaging. Metabolic causes for coma were uncommon. PMID- 15757017 TI - Severe falciparum malaria simulating fulminant hepatic failure. AB - OBJECTIVE: To identify clinical and laboratory features of patients with malarial hepatitis simulating fulminant hepatic failure (MHsFHF) and distinguish it from viral FHF. PATIENTS AND METHODS: At a tertiary care unit in Bangalore, India, we compared clinical and laboratory characteristics of 25 patients with MHsFHF with those of 25 patients with viral FHF from November 1996 to January 2000. RESULTS: No statistically significant differences were seen in duration of jaundice, altered consciousness, and the interval between onset of jaundice and altered consciousness between the 2 groups. Hepatomegaly and splenomegaly were present in 72% and 48% of patients with MHsFHF and in 12% and 0% of patients with viral FHF (P<.001), respectively. The MHsFHF group had a significantly lower hemoglobin level (9.3 g/dL vs 12.9 g/dL), total leukocyte count (9.1 x 10(9)/L vs 18 x 10(9)/L), platelet count (44.8 x 10(9)/L vs 218.6 x 10(9)/L), and transaminases (alanine aminotransferase, 86.2 U/L vs 1230.0 U/L; aspartate aminotransferase, 131.9 U/L vs 720.0 U/L) (P<.001). Thrombocytopenia and elevated serum urea nitrogen were universal in patients with MHsFHF. Prothrombin time was abnormal in all patients with viral FHF and in only 1 patient with MHsFHF. Of patients with MHsFHF, 24% died; of patients with viral FHF, 76% died (P=.02). CONCLUSIONS: In endemic areas, severe malaria should be considered in the differential diagnosis of FHF. Hepatomegaly and normal prothrombin time in the setting of FHF are suggestive of malaria, and a peripheral blood smear should be obtained for diagnostic confirmation. PMID- 15757018 TI - Left heart lesions in patients with Ebstein anomaly. AB - OBJECTIVE: To identify the incidence of left heart abnormalities in patients with Ebstein anomaly, recognizing that left-sided lesions in this patient group have been overlooked. PATIENTS AND METHODS: According to the echocardiography database at the Mayo Clinic in Rochester, Minn, 106 consecutive patients with Ebstein anomaly underwent echocardiography between July 1, 2001, and February 28, 2003. Clinical data as well as electrocardiographic and echocardiographic reports and images were reviewed. RESULTS: Ebstein anomaly was severe in 76 patients (72%). Previous tricuspid valve surgery was reported in 46 patients (43%), and previous closure of an atrial septal defect or patent foramen ovale was reported in 34 patients (32%). Left ventricular (LV) myocardial changes resembling noncompaction occurred in 19 patients (17.9%), LV systolic dysfunction in 7 patients (7%), LV diastolic dysfunction in 34 (36%) of 95 patients, and LV dilatation in 4 patients (4%). Additional left-sided cardiac lesions included mitral valve prolapse in 16 patients (15%), bicuspid aortic valve in 8 (8%), mitral valve dysplasia in 4 (4%), and ventricular septal defect in 8 (8%). Wolff-Parkinson-White syndrome occurred in 22 patients (21%). The QRS axis tended to be different in LV noncompaction with a mean +/- SD axis of 12 degrees +/- 74 degrees vs 36 degrees +/- 66 degrees overall (P=.08). Otherwise, there were no differences in clinical or surgical data between the groups with normal and abnormal LV myocardium. CONCLUSIONS: In patients with Ebstein anomaly, left heart abnormalities involving the myocardium or valves were observed in 39% of patients. Ebstein anomaly should not be regarded as a disease confined to the right side of the heart. PMID- 15757019 TI - New technologies for organ replacement and augmentation. AB - The most common causes of disability and death are diseases of the heart, lungs, liver, kidneys, and pancreas, many of which are potentially treated by organ transplantation. The effect of organ dysfunction and failure will likely grow over time, and patients will increasingly expect "safer" transplants, in particular in cases of "preemptive transplantation." New technologies are being developed in part because of the limited availability of organs, and include transplantation with stem cells, tissue engineering, cloning, and xenotransplantation, which some researchers believe promise ready solutions. Although exciting, none of these approaches alone is likely to address the need for organ replacement. We propose that a melding of these new technologies adapted to the distinct challenges and imperatives of the various organs may address this daunting challenge. PMID- 15757020 TI - The roles of bone mineral density, bone turnover, and other properties in reducing fracture risk during antiresorptive therapy. AB - Osteoporosis is a skeletal disorder characterized by compromised bone strength and increased risk of fracture. Properties related to bone strength include rate of bone turnover, bone mineral density, geometry, microarchitecture, and mean degree of mineralization. These properties (with or without bone density) are sometimes collectively referred to as bone quality. Antiresorptive agents may reduce fracture risk by several separate but interrelated effects on these individual properties. For example, antiresorptive agents have been reported to reduce bone turnover, stabilize or increase bone density, preserve or improve microarchitecture, reduce the number or size of resorption sites, and improve mineralization. Although changes in bone architecture and mineralization are not currently measurable in clinical practice, bone turnover is assessed easily in vivo and affects the other bone properties. Moreover, antiresorptive therapies that produce larger decreases in bone turnover markers together with larger increases in bone mineral density are associated with greater reductions in fracture risk, especially at sites primarily composed of cortical bone such as the hip. Reductions in fracture risk are the most convincing evidence of good bone quality. Data from well-designed randomized clinical trials with up to 10 years of continuous antiresorptive therapy have shown that certain antiresorptive agents effectively reduce fracture risk and (together with extensive preclinical data) suggest no deleterious effects on bone quality. PMID- 15757021 TI - Chronic myeloid leukemia: current application of cytogenetics and molecular testing for diagnosis and treatment. AB - Chronic myeloid leukemia provides an illustrative disease model for both molecular pathogenesis of cancer and rational drug therapy. Chronic myeloid leukemia is a clonal stem cell disease caused by an acquired somatic mutation that fuses, through chromosomal translocation, the abl and bcr genes on chromosomes 9 and 22, respectively. The bcr/abl gene product is an oncogenic protein that localizes to the cytoskeleton and displays an up-regulated tyrosine kinase activity that leads to the recruitment of downstream effectors of cell proliferation and cell survival and consequently cell transformation. Such molecular information on pathogenesis has facilitated accurate diagnosis, the development of pathogenesis-targeted drug therapy, and most recently the application of molecular techniques for monitoring minimal residual disease after successful therapy. These issues are discussed within the context of clinical practice. PMID- 15757022 TI - Diagnosis and management of penicillin allergy. AB - Among patients with a reported history of penicillin allergy, 80% to 90% have no evidence of IgE antibodies to penicillin on skin testing and thus avoid penicillin unnecessarily. Moreover, 97% to 99% of such patients with a penicillin skin test negative to the major and minor determinants can tolerate penicillin without risk of an immediate-type hypersensitivity reaction. A penicillin skin test is valuable for evaluating penicillin allergy in patients who need penicillin or cephalosporin. Assessment of sensitivities to penicillin is important to reduce the unnecessary use of antimicrobial agents such as vancomycin. We review the role of penicillin skin testing for evaluating penicillin allergy and the use of cephalosporin in patients with a history of penicillin allergy. PMID- 15757023 TI - 47-year-old man with fever and headache. PMID- 15757024 TI - Axillary vein thrombosis as a manifestation of rapidly progressive neuropathic arthropathy of the shoulder associated with syringomyelia. AB - Neuropathic arthropathy is a destructive joint disease associated frequently with loss of proprioception. Syringomyelia (with or without Arnold-Chiari malformation) is characterized by slow progression over many years; the joints involved most frequently are the shoulders and elbows. Neuropathic arthropathy of the hand is rare, and clinical features are vasomotor problems, digital ulcers, thickening of the fingers, and painless subluxation of the finger joints. We report an unusual case of neuropathic arthropathy of the shoulder and hands, associated with syringomyelia without Arnold-Chiari malformation. Neuropathic arthropathy of the shoulder was rapidly progressive, and axillary vein thrombosis was the first manifestation; however, neuropathic arthropathy of the hands was unrecognized for 7 years, and painless ulceration of the fingers with spontaneous amputation of the terminal phalanx and Raynaud phenomenon were the initial manifestations. To our knowledge, this is the first report of a patient with this constellation of findings. PMID- 15757025 TI - Emerging medical and surgical management strategies in the evaluation and treatment of intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH) accounts for approximately 10% of all strokes and causes high morbidity and mortality. Rupture of the small perforating vessels of the cerebral arteries is caused by chronic hypertension, which induces pathologic changes in the small vessels and accounts for most cases of ICH; however, amyloid angiopathy and other secondary causes are being seen more frequently with the increasing age of the population. Recent computed tomographic studies have revealed that ICH is a dynamic process with up to one third of initial hemorrhages expanding within the first several hours of ictus. Secondary injury is believed to result from the development of cerebral edema and the release of specific neurotoxins associated with the breakdown products of hemoglobin. Treatment is primarily supportive. Surgical evacuation is the treatment of choice for patients with neurologic deterioration from infratentorial hematomas. Randomized trials comparing surgical evacuation to medical management have shown no benefit of surgical removal of supratentorial hemorrhages. New strategies focusing on early hemostasis, improved critical care management, and less invasive surgical techniques for clot evacuation are promising to decrease secondary neurologic injury. We review the pathophysiology of ICH, its medical management, and new treatment strategies for improving patient outcome. PMID- 15757026 TI - Family-based therapy for adolescents with anorexia nervosa. PMID- 15757027 TI - Family-based therapy for adolescents with anorexia nervosa. PMID- 15757028 TI - Quality care is job one. PMID- 15757029 TI - Appropriate preoperative investigations in the management of rectal cancer. PMID- 15757030 TI - Audit to improve. PMID- 15757031 TI - Single versus sequential fine-needle aspiration biopsy in the management of thyroid nodular disease. AB - Fine-needle aspiration biopsy (FNAB) is considered a safe, reliable and cost effective means of selecting thyroid nodules with risk for malignancy. However, there are limitations of this method including false positive/negative and "nondiagnostic" results that may be reduced by repeating FNAB. OBJECTIVE: To evaluate accuracy, sensitivity, specificity and costs of sequential FNAB in the management of thyroid nodular disease. METHODS: Charts of all patients who underwent thyroidectomy at a university teaching hospital in Toronto from 1998 to 2000 were reviewed. FNAB reports of "suspicious for malignancy," "follicular lesion" and "cellular atypia" were considered to be positive. Data were analyzed with chi2 and z tests. RESULTS: There were 268 patients (225 women and 43 men; age range 18-89 yr; mean age 47 yr) who underwent a total of 449 FNABs (mean 1.7 FNABs/patient) within a year before thyroidectomy. Accuracy (63.8%), sensitivity (73.8%) and specificity (69%) were determined for single FNABs. Sequential FNAB increased the accuracy of method by 22.6%, sensitivity by 13.8% and specificity by 6.2%, with reduction of false positive/negative results by 14.2% and "nondiagnostic" results by 100%. However, the costs of sequential cytology per patient were 70% higher than single FNAB. CONCLUSIONS: Multiple FNABs are unpleasant for patients, but useful in the selection for treatment of patients with thyroid nodular diseases. Although sequential FNAB increases the costs of method, the improvement of precision of FNAB may imply a reduction in overall health-care costs. PMID- 15757032 TI - The investigation of primary rectal cancer by surgeons: current pattern of practice. AB - OBJECTIVE: Selection of the optimal treatment strategy for patients with rectal cancer requires appropriate investigation, but published guidelines provide no clear consensus. We examined the current practice pattern for the investigation of primary rectal cancer by general surgeons in the province of Ontario, Canada. METHODS: A telephone interview was completed by 124 surgeons in Ontario who manage patients with rectal cancer, who indicated the investigations they routinely perform in assessment before treatment. An exploratory cluster analysis was used to identify surgeon-related variables that predicted the pattern of investigation; these were tested in univariate and multivariate analyses. RESULTS: Cluster analysis identified 3 distinct groups of surgeons based on patterns of test usage. Univariate analysis showed that the use of chest radiography, computed tomography of the abdomen and pelvis, and ultrasound of the abdomen varied significantly with the surgeon's subspecialty training, practice location and years in practice. Regression analysis confirmed that each of these 3 variables independently predicted the pattern of preoperative investigation. There were no significant predictors of the use of colonoscopy, sigmoidoscopy or carcinoembryonic antigen level. Over half of surgeons reported that they would have ordered additional imaging tests but did not because of lack of availability. The perceived desirability of additional tests, endorectal ultrasound in particular, varied with training. CONCLUSIONS: The current practice pattern for the preoperative investigation of primary rectal cancer by general surgeons varies significantly with specific surgeon-related variables, with potential impact on the management and outcome of patients with rectal cancer. PMID- 15757033 TI - Personal finances of residents at three Canadian universities. AB - OBJECTIVES: To address 3 research questions (What financial choices do residents make? Are the financial choices of residents similar to those of the general public? Are the financial choices of surgical residents reasonable?), we examined financial data from Canadian residents. METHODS: A written survey was administered to 338 residents (103 of them surgical residents) at 3 Canadian training institutions (University of Toronto, Queen's University and University of Manitoba). Resident household cash flows, assets and liabilities were characterized. Finances for residents were compared with those of the general public, by means of the Survey of Household Spending and Survey of Financial Security. RESULTS: Median resident income was 45,000 dollars annually (Can dollars throughout). With a working spouse, median household income was 87,500 dollars. Among residents, 62% had educational debt (median 37,500 dollars), 39% maintained unpaid credit-card balances (median 1750 dollars), 36% did not budget expenses, 25% maintained cash reserves <275 dollars, and 22% contributed neither to retirement nor nonretirement investments. Residents spent more on vehicles compared with members of the general public (median 17,500 dollars v. 10,720 dollars, p = 0.002) and on monthly housing (median 875 dollars v. 729 dollars, p < 0.001), respectively. Residents were more likely to carry student loans than people in the general population (61% v. 21%), more likely to carry vehicle loans (74% v. 29%) and less likely to carry credit-card debts (39% v. 50%, respectively). Surgical residents had income expectations after graduation higher than current billings justified. Fewer surgical (69%) than anesthesiology residents (88%, p < 0.05) contributed to Registered Retirement Savings Plans. CONCLUSIONS: From this limited sample, residents spend more than age- and income matched members of the general public. Many residents save too little, fail to budget, and carry high educational and credit-card debts. Surgical residents' expectations of future income may be unrealistic. Further study is warranted. PMID- 15757034 TI - A comparison of plate versus staple-and-cast fixation in maintaining femoral tibial alignment after valgus tibial osteotomy. AB - PURPOSE: To compare 2 methods of fixation for maintenance of alignment during healing of valgus tibial osteotomies. METHODS: We performed a retrospective chart and radiographic review of valgus tibial osteotomy cases of staple fixation supplemented by a postoperative cast and of blade plate fixation for maintenance of femoral-tibial alignment during healing of the osteotomy. RESULTS: Both groups (staple-and-cast, n = 16; plate, n = 28) were similar in terms of age, preoperative alignment, extent of osteoarthritis and degree of intra-operative correction. Between-group differences in the maintenance of femoral-tibial alignment during healing were not significant. In both groups there was a strong correlation between degree of bone contact at the osteotomy site and maintenance of alignment (p < 0.005). In cases done with the plate, 90% of osteotomies with good or excellent bone contact maintained alignment during healing; with poor or fair bone contact, 75% had loss of alignment > 5 degrees during healing. There was a trend toward a greater incidence of delayed or non-union with plate fixation compared with staple fixation that did not reach statistical significance. All of these cases of delayed/non-union had loss of femoral-tibial alignment > 5 degrees during healing. CONCLUSION: As a result of this study, we have modified our surgical technique. We now use intra-operative fluoroscopy for optimization of bone contact, and we have reverted to the less invasive staple method for fixation of tibial osteotomies. PMID- 15757036 TI - Surgical education and self-assessment program (SESAP). Category 1, item 10. PMID- 15757035 TI - Defining medical error. AB - Medical errors represent a serious public health problem and pose a threat to patient safety. As health care institutions establish "error" as a clinical and research priority, the answer to perhaps the most fundamental question remains elusive: What is a medical error? To reduce medical error, accurate measurements of its incidence, based on clear and consistent definitions, are essential prerequisites for effective action. Despite a growing body of literature and research on error in medicine, few studies have defined or measured "medical error" directly. Instead, researchers have adopted surrogate measures of error that largely depend on adverse patient outcomes or injury (i.e., are outcome dependent). A lack of standardized nomenclature and the use of multiple and overlapping definitions of medical error have hindered data synthesis, analysis, collaborative work and evaluation of the impact of changes in health care delivery. The primary objective of this review is to highlight the need for a clear, comprehensive and universally accepted definition of medical error that explicitly includes the key domains of error causation and captures the faulty processes that cause errors, irrespective of outcome. PMID- 15757037 TI - Surgical images: soft tissue. Incarcerated gallbladder in a parastomal hernia. PMID- 15757038 TI - Radiology for the surgeon. Musculoskeletal case 34. Osteonecrosis and myopathy of skeletal muscle. PMID- 15757040 TI - Users' guide to the surgical literature. Self-audit and practice appraisal for surgeons. PMID- 15757041 TI - Canadian Association of General Surgeons evidence based reviews in surgery. 12. Primary repair for penetrating colon injuries. PMID- 15757039 TI - Emergent management of pelvic ring injuries: an update. PMID- 15757042 TI - Dysplasia epiphysealis hemimelica of the ankle in children. PMID- 15757043 TI - Open tracheostomy in a suspect severe acute respiratory syndrome (SARS) patient: brief technical communication. PMID- 15757044 TI - Elective and emergency surgery in patients with severe acute respiratory syndrome (SARS). PMID- 15757045 TI - An unusual presentation of a secondary extramedullary plasmacytoma in a patient with multiple myeloma. PMID- 15757046 TI - Laparoscopic management of incarcerated obturator hernia. PMID- 15757047 TI - Retroperitoneal hemorrhage secondary to a ruptured cavernous hemangioma. PMID- 15757048 TI - Management of perforated duodenal diverticula. PMID- 15757049 TI - Efficacy of laparoscopic appendectomy in appendicitis with peritonitis. AB - Laparoscopic appendectomy (LA) is safe and effective in cases of peritonitis, perforation, and abscess. We investigated our conversion rate and clinical outcomes in this patient population, as well as preoperative factors that predict operative conversion. A retrospective nonrandomized cohort of 92 patients underwent LA for acute appendicitis with peritonitis, perforation, or abscess at our institution between 1997 and 2002. Thirty-six of the 92 were converted to open appendectomy (OA), yielding a conversion rate of 39 per cent. The presence of phlegmon (42%), nonvisualized appendix (44%), technical failures (8%), and bleeding (6%) were reasons for conversion. Preoperative data had no predictive value for conversion. CT scan findings of free fluid, phlegmon, and abscess did not correlate with findings at the time of surgery. Total complication rates were 8.9 per cent in the LA group as compared to 50 per cent in the converted cohort. Postoperative data showed LA patients stayed 3.2 days versus 6.9 days for converted patients (P = 0.01). LA patients had less pneumonia (P = 0.02), intra abdominal abscess (P = 0.01), ileus (P = 0.01), and readmissions (P = 0.01). LA is safe and effective in patients with appendicitis with peritonitis, perforation, and abscess, resulting in shorter hospital stays and less complication. PMID- 15757050 TI - Laparoscopic revision of bariatric procedures: is it feasible? AB - Reoperative bariatric surgery is required in 10 per cent to 20 per cent of patients secondary to weight regain or complications of the previous procedure. This study evaluates the feasibility of performing the revision procedure laparoscopically. A retrospective review of all patients undergoing revision of a previous weight loss procedure between October 1998 and November 2003 was conducted. Demographics, indications for surgery, operative findings, and complications were reviewed. Thirty-nine revisions were performed in 37 patients. Indications for revision were failure to lose weight (22), gastric outlet stricture (10), refractory gastroesophageal reflux (GERD) (6), and blind loop syndrome (1). All 39 procedures were revised to Roux-en-Y gastric bypass (RYGBP), with 18 open revisions (OR) and 21 laparoscopic revisions (LR). Ten of the 21 LR (48%) were converted to an open procedure due to adhesions or unclear anatomy. Early complications requiring operation were noted in five procedures (two OR, three LR). Nine patients (seven OR, two LR) required surgery at least 3 months following their revision. One patient died (LR). The difference in body mass index (kg/m2) (BMI) pre- and post-op was 43.3+/-9.9 versus 37.4+/-9.2, P = 0.01 (follow-up 5 months), but no significant BMI differences between LR and OR patients were seen. Revisional bariatric surgery is associated with more complications requiring surgery early in the laparoscopic population versus more late complications in those approached open. Revisional bariatric surgery can be approached laparoscopically and with acceptable morbidity comparable to patients whose revision is approached open. PMID- 15757051 TI - Correlation between perceptual, visuo-spatial, and psychomotor aptitude to duration of training required to reach performance goals on the MIST-VR surgical simulator. AB - Given the dynamic nature of modern surgical education, determining factors that may improve the efficiency of laparoscopic training is warranted. The objective of this study was to analyze whether perceptual, visuo-spatial, or psychomotor aptitude are related to the amount of training required to reach specific performance-based goals on a virtual reality surgical simulator. Sixteen MS4 medical students participated in an elective skills course intended to train laparoscopic skills. All were tested for perceptual, visuo-spatial, and psychomotor aptitude using previously validated psychological tests. Training involved as many instructor-guided 1-hour sessions as needed to reach performance goals on a custom designed MIST-VR manipulation-diathermy task (Mentice AB, Gothenberg, Sweden). Thirteen subjects reached performance goals by the end of the course. Two were excluded from analysis due to previous experience with the MIST-VR (total n = 11). Perceptual ability (r = -0.76, P = 0.007) and psychomotor skills (r = 0.62, P = 0.04) significantly correlated with the number of trials required. Visuo-spatial ability did not significantly correlate with training duration. The number of trials required to train subjects to performance goals on the MIST-VR manipulation diathermy task is significantly related to perceptual and psychomotor aptitude. PMID- 15757052 TI - Accuracy of intraoperative gross examination of surgical margin status in women undergoing partial mastectomy for breast malignancy. AB - Margin status is an important prognostic factor for local recurrence after partial mastectomy for breast malignancy. Options for intraoperative evaluation of margin status include gross examination of the specimen, frozen section, and "touch preparation" cytology. This study evaluates the accuracy of gross examination without other intraoperative pathological analysis as a method of determining margin status. Records of 254 consecutive patients undergoing partial mastectomy for 255 breast malignancies (199 invasive, 56 DCIS) over 6 years were analyzed retrospectively. All women underwent en bloc excision of the primary lesion with gross examination of margin status by the surgeon and pathologist. All suspicious areas were reexcised, and the specimen was inked, serially sectioned at 2-3 mm intervals and examined with hematoxylin and eosin (H&E) stains. Specimens with tumor <2 mm from a margin were considered margin-positive and those with all tumor > or =2 mm from the margin were designated margin negative. One hundred fourteen (45%) of the 255 segmental resections were considered to have grossly tumor-free margins, and intraoperative reexcision was not performed. Ninety-six (84%) of these specimens had histologically negative margins. Gross examination prompted intraoperative reexcision in 141 (55%) cases. Ninety-five (67%) of these 141 resections had tumor-free margins on histopathology. Overall, the final margin was involved in 64 of the 255 partial mastectomies. Seventeen (27%) women with initially margin-positive resections underwent mastectomy, while 46 (72%) underwent reexcision, which was margin negative in 41 (89%). After a median follow-up of 42 months, there have been eight (3.5%) local recurrences. The initial margin-positive rate was similar in ductal carcinoma in situ (DCIS) (30%) and invasive carcinoma (24%). Margin status was correlated with nodal status; there was no correlation with age, tumor size, grade hormone receptor status, or type of diagnostic biopsy. Gross examination of the resection specimen does not reflect margin status in at least 25 per cent of women undergoing partial mastectomy for breast malignancy. Other techniques for evaluation of margin status should be considered to reduce the need for reexcision of involved margins. We are currently designing a prospective clinical trial to examine the efficacy of new techniques for intraoperative evaluation of margin status. PMID- 15757053 TI - Determining standards for laparoscopic proficiency using virtual reality. AB - Laparoscopic training using virtual reality has proven effective, but rates of skill acquisition vary widely. We hypothesize that training to predetermined expert levels may more efficiently establish proficiency. Our purpose was to determine expert levels for performance-based training. Four surgeons established as laparoscopic experts performed 11 repetitions of 12 tasks. One surgeon (EXP-1) had extensive Minimally Invasive Surgical Trainer-Virtual Reality (MIST VR) exposure and formal laparoscopic fellowship training. Trimmed mean scores for each were determined as expert levels. A composite score (EXP-C) was defined as the average of all four expert levels. Thirty-seven surgery residents without prior MIST VR exposure and two research residents with extensive MIST VR exposure completed three repetitions of each task to determine baseline performance. Scores for EXP-1 and EXP-C were plotted against the best score of each participant. On average, the EXP-C level was reached or exceeded by 7 of the 37 (19%) residents. In contrast, the EXP-1 level was reached or exceeded by 1 of 37 (3%) residents and both research residents on all tasks. These data suggest the EXP-C level may be too lenient, whereas the EXP-1 level is more challenging and should result in adequate skill acquisition. Such standards should be further developed and integrated into surgical education. PMID- 15757054 TI - A comparison of continuous renal replacement therapy to intermittent dialysis in the management of renal insufficiency in the acutely III surgical patient. AB - Acute renal failure (ARF) occurs in 10 per cent to 23 per cent of intensive care unit patients with mortality ranging from 50 per cent to 90 per cent. ARF is characterized by an acute decline in renal function as measured by urine output (UOP), serum creatinine, and blood urea nitrogen (BUN). Causes may be prerenal, intrarenal, or postrenal. Treatment consists of renal replacement therapy (RRT), either intermittent (ID) or continuous (CRRT). Indications for initiation of dialysis include oliguria, acidemia, azotemia, hyperkalemia, uremic complications, or significant edema. Overall, the literature comparing CRRT to ID is poor. No studies of only surgical/trauma patients have been published. We hypothesize that renal function and hemodynamic stability in trauma/ surgical critical care patients are better preserved by CRRT than by ID. We performed a retrospective review of trauma/surgical critical care patients requiring renal supportive therapy. Thirty patients received CRRT and 27 patients received ID. The study was controlled for severity of illness and demographics. Outcomes assessed were survival, renal function, acid-base balance, hemodynamic stability, and oxygenation/ventilation parameters. Populations were similar across demographics and severity of illness. Renal function, measured by creatinine clearance, was statistically greater with CRRT (P = 0.035). There was better control of azotemia with CRRT: BUN was lower (P = 0.000) and creatinine was lower (P = 0.000). Mean arterial blood pressure was greater (P = 0.021) with CRRT. No difference in oxygenation/ventilation parameters or pH was found between groups. CRRT results in an enhancement of renal function with improved creatinine clearance at the time of dialysis discontinuation. CRRT provides better control of azotemia while preserving hemodynamic stability in patients undergoing renal replacement therapy. Prospective randomized controlled studies and larger sample sizes are needed to further evaluate these modalities. PMID- 15757055 TI - Evaluation of the clinical pathway for laparoscopic cholecystectomy. AB - Clinical pathways are comprehensive systematized patient care plans for specific procedures. The clinical pathway for laparoscopic cholecystectomy was implemented in our department in March 2002. The aim of this study is to evaluate the clinical pathway for this procedure 1 year after implementation. A study was conducted on all the patients included in the clinical pathway since its implementation. The assessment criteria include degree of compliance, indicators of clinical care effectiveness, financial impact, and survey-based indicators of satisfaction. The results are compared to a series of patients undergoing surgery the year prior to implementation of the clinical pathway. As our hospital has a system of cost management, we analyzed the mean cost per procedure before and after clinical pathway implementation. Evaluation was made of a series of 160 consecutive patients who underwent surgery during the period 1 year prior to development of the clinical pathway and met the accepted inclusion criteria. The mean length of hospital stay was 3.27 days, and the mean cost per procedure before pathway implementation was 2149 (+/-768) euros. One year after implementation of the pathway, 140 patients were included (i.e., an inclusion rate of 100%). The mean length of hospital stay of the patients included in the clinical pathway was 2.2 days. The degree of compliance with stays was 66.7 per cent. The most frequent reasons for noncompliance were staff-dependent, followed by patient-dependent causes (oral intolerance, pain, etc.). The mean cost in the series of patients included in the clinical pathway was 1845 (+/-618) euros. Laparoscopic cholecystectomy is an ideal procedure for commencing the systemization of clinical pathways. Results show that it has significantly reduced the length of hospital stay and mean cost per procedure with no increased morbidity and with a high degree of patient satisfaction. PMID- 15757056 TI - Selective use of cardiopulmonary bypass in trauma patients. AB - The need for cardiopulmonary bypass (CPB) in the treatment of trauma patients is controversial, and not all level I trauma centers have CPB readily available. Our purpose was to review the selective use of CPB in the management of trauma victims at a level I trauma center in Los Angeles County. We reviewed the records of all patients for whom the CPB team was called in from 1994 to 2002. Perfusionists were present for the initial operative management of 24 patients, 22 (92%) of which were male. Twelve patients had penetrating and nine had blunt injuries, two were severely hypothermic, and the last suffered embolization of a bullet to the pulmonary artery. Overall survival was 75 per cent. Sixteen (67%) patients required CPB due to the life-threatening nature of their injuries and/or hemodynamic instability; 11 (69%) survived. The remaining 8 patients were operated on with the CPB team present but on standby; 7 (88%) survived. Cardiopulmonary bypass could be life-saving in select trauma patients with major chest injuries. Hypothermia, acidemia, and shock can be reversed earlier while allowing increased time to gain adequate exposure and perform quality repairs. Level I trauma centers should have CPB capabilities immediately available. PMID- 15757057 TI - Routine chest X-ray after percutaneous tracheostomy is unnecessary. AB - Percutaneous tracheostomy (PT) is an increasingly common procedure in the management of critically ill patients. Current practice for both open and percutaneous tracheostomies is a post-procedure chest X-ray to rule out potentially life-threatening complications such as a pneumothorax or tube malposition. Our study evaluated the utility of chest X-ray after PT. A retrospective chart review was conducted for patients undergoing PT at Kern Medical Center between January 1999 and December 2003. Charts were reviewed for age, sex, and clinical outcome as well as the radiologist's interpretation of the postprocedure chest X-ray. A total of 73 procedures were completed in 47 men and 26 women. The majority of the tracheostomies were in trauma patients who needed prolonged ventilatory support. There were no complications identified on postprocedure chest X-ray. A single patient was converted to an open procedure secondary to bleeding. We conclude that routine chest X-ray after PT is unnecessary. PMID- 15757058 TI - Association of scapular fractures and blunt thoracic aortic injury: fact or fiction? AB - It is classically taught that scapular fractures (SF) are commonly associated with blunt thoracic aortic injury (BTAI). The purpose of this study was to determine the association between SF and BTAI. A 10-year retrospective review of blunt trauma admissions from two level I trauma centers located in different geographic regions, Washington Hospital Center (WHC) and Los Angeles County Medical Center and the University of Southern California (LAC/USC), was performed. Patients with SF and BTAI were identified, and records were reviewed to determine associated injuries. We identified 35,541 blunt trauma admissions (WHC: 12,971, LAC/USC: 22,570). SF and BTAI occurred in 1.1 per cent and 0.6 per cent of patients, respectively. Most of the patients with SF had associated injuries (99%). Only four patients with SF had BTAI (4/392; 1.0%). The most common injuries associated with SF were rib (43%), lower extremity (36%), and upper extremity (33%) fractures. SF is uncommon after blunt trauma. Patients with SF almost always have significant associated injuries. Although SF indicates a high amount of energy transmitted to the upper thorax, these patients rarely have BTAI. SF should not be used as an indicator of possible BTAI. PMID- 15757059 TI - Umbilical hernia repair in cirrhotic patients: utility of temporary peritoneal dialysis catheter. AB - Umbilical herniorrhaphy in cirrhotic patients with ascites is associated with a significant morbidity, recurrence rate, and mortality and therefore is often managed expectantly. Operative repair is indicated if an ascites leak or infection develops. Surgeons must consider the management of postoperative ascites to reduce recurrence rates and complications. We present a unique method using temporary peritoneal dialysis catheter placement (PD). Eight patients with moderate to massive ascites underwent umbilical herniorrhaphy with concomitant peritoneal dialysis placement. Patients have been followed for 8 to 30 months. All patients had successful repair of their hernia with 1 recurrence at 6 months and 1 late death (14 months). Patients were able to effectively control ascites using the PD catheter at home. There were no postoperative infections. The placement of a temporary PD catheter during umbilical herniorrhaphy provides a method for effective control of ascites in patients with cirrhosis. The technique has several advantages including outpatient management during the postoperative period and for easy removal of the catheter when no longer needed. PMID- 15757060 TI - Embolization of an acute renal arteriovenous fistula following a stab wound: case report and review of the literature. AB - Surgery has traditionally been the definitive form of invasive management for renal vascular injuries. There is a growing trend in the use of endovascular techniques as an alternative to surgery in the trauma setting. We present the case of a 24-year-old woman with an acute renal arteriovenous fistula caused by a stab wound in the left flank, which was successfully managed with selective arterial embolization. This represents only the second reported case of such an approach in the acute setting. PMID- 15757061 TI - Evidence-based medicine applied to sentinel lymph node biopsy in patients with breast cancer. AB - Sentinel lymph node biopsy (SLNB) has not been examined using the principles of evidence-based medicine (EBM). Specifically, likelihood ratios have not been used to assess the validity of SLNB. The Surveillance, Epidemiology, and End Results (SEER) public database of the National Cancer Institute was used to establish the baseline or pretest probability of finding a positive lymph axillary node for each stage of breast cancer. Rates of false negative results of SLNB for all breast cancer stages were determined from the surgical literature. Positive and negative likelihood ratios (LR) were calculated. For each stage of breast cancer, the Bayesian nomogram was used to find the post-test probability of missing a metastatic axillary node when the SLN was negative. The SEER database of 213,292 female patients with breast cancer yielded the following rates of positivity of axillary lymph nodes for each breast tumor size: T1a, 7.8 per cent; T1b, 13.3 per cent; T1c, 28.5 per cent; T2, 50.2 per cent; T3, 70.1 per cent. The combined data from 13 published studies of SLNB (6444 successful SLNBs) demonstrated a false negative rate of 8.5 per cent. The LR of a negative test is 0.086. According to the nomogram, the chances of missing a positive node for stage of cancer are as follows: T1a, 0.7 per cent; T1b, 1.5 per cent; T1c, 3.0 per cent; T2, 7 per cent; T3, 18 per cent. The risk of missing a positive axillary node can accurately be estimated for each stage of breast cancer using the LR, which is much more useful than the simple false negative rate. Surgeons should use this information when deciding whether to perform SLNB and in their informed consent discussions. PMID- 15757062 TI - Herbal product use by hispanic surgical patients. AB - We hypothesized that the use of herbal products is highly prevalent in Hispanic surgical patients on both sides of the U.S./Mexico border. One hundred fifteen patients were interviewed in El Paso and 112 in Ciudad Juarez from December 2001 to June 2002. This prospective, randomized study evaluated patients aged 18 years or older that were scheduled for surgery. Sixty-two per cent of our sample in the United States and 81 per cent of patients in Mexico reported using herbal products in the past year. About 58 per cent of patients in the United States and 49 per cent in Mexico rated the products as "excellent" in treating their conditions. Ninety-two percent of U.S, and 93 per cent of Mexican patients did not inform their physician of their herbal use. Preoperative assessment of patients did not include inquiries about herbal products in either hospital. With the growing use of herbal products and because most users do not inform their physician, there is increasing concern related to the possible occurrence of herbal-related adverse events during and after surgery. The fact that health care personnel did not inquire about herbal product use before surgery is an added concern. PMID- 15757063 TI - Areolar-cutaneous junction injection boosts activity in sentinel node by more than 50 times compared to perilesional injection: implications for morbidity reduction. AB - The combination of concurrently administered perilesional radiotracer injections and areolar-cutaneous junction radiotracer injections during one imaging session in the detection of sentinel nodes (SN) in breast cancer is new. A case is presented where the perilesional injections of radiotracer produced a faint node. Subsequently, 45 minutes later, the same patient received an injection of radiotracer at the areolar-cutaneous junction, which "boosted" the activity in the original SN by more than 50 times. This case illustrates the marked improvement in SN counts that can occur with the addition of areolar-cutaneous junction injections to perilesional injections. The perilesional component of this hybrid injection technique maintains the ability to visualize internal mammary and extra-axillary SN. Even more importantly, these "hotter" nodes have significant implications for morbidity reduction, the main goal of sentinel lymph node biopsy in itself. PMID- 15757064 TI - TFC (thoracic fluid content): a new parameter for assessment of changes in chest fluid volume. AB - Clinical assessment of cardiac status can be difficult and incomplete without an assessment of fluid volume status, especially of the lungs. Now, a new parameter is available, thoracic fluid content (TFC). It is an indicator of total fluid volume, both intracellular and extracellular. Because it is measured noninvasively using impedance cardiography (ICG), it could be a welcome addition to the physician's assessment. An evaluation of TFC was performed beginning with 1) an examination of chest impedance (Z) as an accurate means of following fluid changes, 2) the relationship of TFC to Z using both loop and spot electrodes, and 3) clinical applications of TFC. In 1) 12 dogs, Z was superior (r = 0.935, P < 0.006) to 10 traditional hemodynamic and gas transfer parameters in trending a lactated Ringer's infusion; 2) a plastic model, changes in TFC values derived from Z measurements using both loop and spot electrodes were virtually identical and paralleled infused saline (r = 0.999, P < 0.001); 3) the clinical setting, TFC trended fluid changes well. From these results, we conclude that TFC is a reliable measurement of chest fluid status and of changes in that fluid. Along with cardiac index (CI), also provided by the ICG monitor, TFC can be very helpful to the clinician. PMID- 15757065 TI - Gallbladder volvulus: report of two original cases and review of the literature. AB - Gallbladder volvulus is defined as the rotation of the gallbladder on its mesentery along the axis of the cystic duct and cystic artery. It is an unusual and rare occurrence with a predilection for women in their seventh or eight decades of life. Only about 300 cases have been reported in the literature ranging from ages 2 to 100 years old. The etiology of gallbladder volvulus remains speculative; however, the presence of a redundant mesentery is a prerequisite for torsion. Gall-bladder volvulus leads to occlusive obstruction of biliary drainage and blood flow. With early diagnosis and surgical intervention, the disease maintains a low mortality of approximately 5 per cent. We present two original cases of acute gallbladder volvulus successfully treated with cholecystectomy, as well as a review of the literature. PMID- 15757066 TI - Adrenocortical carcinoma: a single institution experience. AB - Adrenocortical carcinoma (ADCC) ranks among the least common malignant endocrine tumors. Surgical resection is considered the most important treatment for this neoplasm. Medical records of patients with the diagnosis of ADCC between 1990 and 2000 were reviewed. Patient and pathologic factors were analyzed with overall survival as the primary endpoint. Statistical analysis was performed by the method of Kaplan-Meier. There were a total of 17 patients, with a mean age of 56 years. Twelve per cent presented as an asymptomatic mass, 41 per cent as a functional tumor, and 47 per cent as a nonfunctioning tumor. Primary treatment was surgical resection in 71 per cent. There was no operative mortality and one complication. Seven patients presented with stage II, five with stage III, four with stage IV, and in one could not be determined. Median follow-up was 12.8 months, median survival 67, 13, and 3 months for stages II, III, and IV, respectively. Older age, distant metastasis, nonoperative management, positive margins, advanced tumor stage, and venous invasion were significantly associated with worse overall actuarial survival. Survival for ADCC is poor. Factors associated with a worse prognosis were stage of disease, nonoperative management, positive surgical margins, vascular invasion, and older age. PMID- 15757067 TI - CMS issues pressure ulcer guidelines. PMID- 15757068 TI - Emergency preparedness keeps danger at bay. PMID- 15757069 TI - Establishing corporate objectives. PMID- 15757070 TI - The importance of oral health. PMID- 15757071 TI - Medicare to cover brain scans. PMID- 15757072 TI - Data's role in quality improvement. PMID- 15757073 TI - Another pill for weight reduction. PMID- 15757074 TI - Congenital coronary artery anomalies in the adult: a new practical viewpoint. AB - There is a certain confusion as to the universal classification of coronary artery anomalies (CAAs), since the existing classifications are often fragmented and difficult to apply as they combine many anatomical, angiographic, and clinical elements. A practical global classification of CAAs for diagnostic and clinical purposes has not been proposed previously. The authors propose to describe CAAs on the basis of seven angiographic patterns (1: hypoplasia/atresia, 2: hyperdominance, 3: fistula, 4: originating from other arteries, 5: originating from the wrong sinus, 6: splitting, and 7: tunneling) and four clinical significance classes (benign--class A, relevant--class B, severe--class C, and critical--class D). This global classification can be applicable to almost all significantCAAs, and in the authors' view it might help both angiographers and clinical cardiologists to familiarize themselves rapidly with a comprehensive description of most CAAs by simplifying the transmission of the angiographic data. PMID- 15757076 TI - A case of tamponade complicating pregnancy. PMID- 15757075 TI - The effects of statin therapy on plasma markers of inflammation in patients without vascular disease. AB - Statins may reduce cardiovascular risk via mechanisms other than cholesterol reduction alone. This decrease in risk may in part be secondary to pleiotropic effects such as decreased inflammation and improved endothelial function. Statin therapy has been shown to affect the levels of certain plasma markers of inflammation, markers which are predictive of cardiovascular risk and may also play a role in the disease process. This article will review the relationship between inflammatory markers and cardiovascular disease risk and the effects of statins on these markers in subjects without a known history of coronary disease. PMID- 15757077 TI - Effect of preexisting statin use on expression of C-reactive protein, adhesion molecules, interleukin-6, and antioxidized low-density lipoprotein antibody in patients with unstable angina undergoing coronary stenting. AB - BACKGROUND: Statins are believed to reduce coronary heart disease by mechanisms in addition to their well-known cholesterol lowering effect. HYPOTHESIS: We studied the effect of statins on expression of C-reactive protein (CRP), interleukin-6 (IL-6), adhesion molecules, and antioxidized low-density lipoprotein antibody (anti-oxLDL Ab) in patients with unstable angina (Braunwald class IIb or IIIb) undergoing coronary stenting. METHODS: Consecutive 50 patients with unstable angina were included in the study. We classified the study subjects as patients using statins (Group A, n=20, men 10, mean age 62 years) and patients not using statins (Group B, n=30, men 15, mean age 60 years). RESULTS: Baseline levels of inflammatory markers were similar in the two groups. However, 24 h after coronary stenting, serum levels of CRP (2.00 vs. 4.63 mg/l, p < 0.05), intercellular adhesion molecule-1 (ICAM-1) (217 vs. 261 ng/ml, p < 0.01), and anti-oxLDL Ab (8.97 vs. 12.96 U/ml, p < 0.05) were significantly higher in Group B than in Group A. Furthermore, 72 h after coronary stenting, serum levels of CRP (3.00 vs. 5.50 mg/l, p < 0.01) and ICAM-1 (222 vs. 277 ng/ml, p < 0.05) were significantly higher in Group B than in Group A. CONCLUSIONS: Preexisting statin therapy plays a role in reducing the serum levels of CRP, ICAM-1, and anti-oxLDL Ab after coronary stenting in patients with unstable angina. These data support an anti-inflammatory or plaque-stabilizing effect of statin therapy. PMID- 15757078 TI - The new definition of myocardial infarction--can we use it? AB - BACKGROUND: A joint committee of the European Society of Cardiology and the American College of Cardiology (ESC/ACC) recently redefined myocardial infarction. HYPOTHESIS: The objective of this study was to examine the outcome of diagnoses from more than 500 patients admitted to a university hospital coronary care unit (CCU), when the ESC/ACC committee cut-off levels were compared with the Swedish diagnostic criteria for acute myocardial infarction (MI), comparable with everyday practice in most countries. METHODS: Creatine kinase-MB, troponin I, and troponin T were measured in 525 patients admitted consecutively to the CCU, Huddinge University Hospital, with possible myocardial ischemia lasting <12 h before arrival. RESULTS: The ESC/ACC definition of MI increased the number of MIs by 3-32% compared with the number achieved when Swedish diagnostic criteria for acute MI were used. A significant number of patients with elevated cardiac enzymes presented with acute heart failure, tachycardia, pulmonary embolism, and sepsis as initial symptom. CONCLUSIONS: In this study of more than 500 patients with possible myocardial ischemia admitted consecutively to the CCU at a university hospital, the ESC/ACC definition of MI increased the number of MIs by 3-32% compared with the number achieved when Swedish diagnostic criteria for acute MI were used. A majority of the patients identified with ESC/ACC cut-off levels presented with myocardial ischemia as the primary symptom, whereas many of the other patients had acute heart failure and tachycardia as initial symptom. It is unclear whether patients in this latter group should be labelled as having MI; there are no clinical studies providing guidance in this situation. PMID- 15757079 TI - Coronary artery calcium by digital cinefluoroscopy in patients with pain suggestive of an acute coronary syndrome. AB - BACKGROUND: Detection of coronary calcium may be a useful noninvasive approach for detecting coronary artery disease (CAD) in subjects presenting to the emergency department with chest pain. HYPOTHESIS: We tried to assess the diagnostic value of coronary artery calcium (CAC) detection by digital cinefluoroscopy in patients with new-onset chest pain suggestive of an acute coronary syndrome. METHODS: In 97 consecutive patients (70 men, 27 women, mean standard deviation [SD] age 55 (11) and 60 (8) years, respectively), with new onset chest pain suggestive of an acute coronary syndrome, nondiagnostic electrocardiogram, and normal initial creatine kinase (CK)-MB, digital cinefluoroscopy was performed for CAC detection. RESULTS: All patients underwent routine clinical evaluation with treadmill exercise test, thallium scintigraphy, dobutamine stress echocardiography, and coronary angiography, as needed clinically and blinded to the cinefluoroscopy results. Coronary artery calcium was present in 27 of 33 (81.8%) of patients with and in 10 of 64 (15.6%) of patients without CAD, p < 0.0001. The presence of CAC had 82% sensitivity, 84% specificity, 73% positive predictive value, and 90% negative predictive value for CAD diagnosis (odds ratio = 24.3, 95% confidence interval 7.98-73.94). CONCLUSIONS: In patients with acute chest pain, nondiagnostic electrocardiogram and normal initial enzyme evaluation, CAC detection by digital cinefluoroscopy appears to have high sensitivity, specificity, and negative predictive value for CAD diagnosis. PMID- 15757081 TI - Abnormal myocardial perfusion and contractile recruitment during exercise in type 1 diabetic patients. AB - BACKGROUND: No data are available on the relationship between myocardial perfusion and left ventricular (LV) function in type 1 diabetes mellitus (T1DM), which may constitute a factor explaining the progressive contractile dysfunction to the overt phase of diabetic cardiomyopathy. HYPOTHESIS: This study was undertaken to test whether myocardial perfusion abnormalities are present at rest and during exercise and whether they are related to contractile dysfunction in T1DM. METHODS: Twenty-two patients with T1DM, aged 32 +/- 8.3 years, without macro- or microvascular complications, and 10 controls, aged 31 +/- 3 years, were studied. Left ventricular function and myocardial perfusion were assessed by two dimensional and myocardial contrast echocardiography at rest and during handgrip (HG). RESULTS: Fourteen patients with T1DM showed a decline in LV ejection fraction (LVEF) during HG (Group 1) while 8 had a normal response (Group 2). Both basal myocardial blood volume (MBV) and velocity (beta) were normal in T1DM. During exercise, MBV and beta increased and were associated with an increase in myocardial blood flow (MBF) in controls. In T1DM, beta did not change and MBV increased only in Group 2, while this increase was not observed in Group 1 (controls: 14.9 +/- 2.3 vs. Group 1: 7.6 +/- 1.6, p < 0.001; and vs. Group 2: 10.2 +/- 2.8, p < 0.001), beta (0.86 +/- 0.12 vs. 0.68 +/- 0.14, p < 0.001; and vs. 0.67 +/- 0.15, p < 0.001). A correlation between the ratio exercise MBF/resting MBF and LVEF at peak exercise in T1DM was observed (r = 0.805, p < 0.001). CONCLUSIONS: A large proportion of patients with T1DM exhibit abnormalities in myocardial adaptable capacity to match an acute overload, which are related to a defective increase in myocardial perfusion. PMID- 15757080 TI - Cardiovascular anomalies in children and young adults with Ullrich-Turner syndrome the Erlangen experience. AB - BACKGROUND: Females with Ullrich-Turner syndrome (UTS) have typical clinical features such as short stature, ovarian failure, visible dysmorphic stigmata, and abnormalities in different organs such as kidney or heart. HYPOTHESIS: The aim of the present study was to analyze the distribution, prevalence, and relative risk of cardiovascular anomalies (CVA) in females with Ullrich-Turner syndrome (UTS) seen at one single center compared with that of the regional Bavarian population. METHODS: The associations between CVA and karyotype were determined. In all, 117 girls and women with UTS, aged between 3 and 43 years (median 17.4 years) were studied retrospectively. The detailed cardiologic status including echocardiography was available in all patients. The prevalences of each cardiovascular anomaly were determined. On the basis of published epidemiologic data of CVA in Bavarian children, we assessed the relative risks of each CVA. RESULTS: Thirty-five (29.9%) girls with UTS had at least one CVA. In all of these CVAs, coarctation of the aorta and bicuspid aortic valve occurred most often (18.5% each). The aortic malformations represented over two-thirds of all CVA (72.8%), whereas anomalies of the septum (8.6%), mitral valve (6.2%), pulmonary veins (4.9%), and other locations together accounted for the other third. Bicuspid aortic valve and partial anomalous pulmonary venous drainage were associated with the highest relative risk (RR) (3603 and 1293, respectively) compared with the Bavarian population. The overall RR of CVA was 48.7. Of the 117 girls and women examined, 64 (54.7%) had complete monosomy 45 X. CONCLUSIONS: Our data demonstrate that about every third female with UTS is affected with at least one CVA, mainly left sided and associated with aortic structures. Our results underline the necessity of thorough cardiologic evaluation. PMID- 15757082 TI - Three-dimensional computed tomography imaging of left atrial anatomy for atrial fibrillation ablation. PMID- 15757083 TI - Michael B. Gravanis. PMID- 15757084 TI - [Software and hardware for computer equipment designed for psychophysiological examination]. AB - Principles of designing software and hardware for new computer equipment for psychophysiological examination were elaborated on the basis of theoretical prerequisites and of analysis of the use of equipment manufactured serially for quantitation of indices of the human higher nervous activity (HNA). The hierarchic structuring of software and hardware enabled, through unification of solutions, the development of new modifications of two equipment sets: computer based unit "Psihomat" KPFK-99 for psychophysiological examination and computer based unit "STABILOTEST" ST-01 for the evaluation of the central nervous system by the stability parameters in maintaining the vertical posture. The units have new improved functional features and provide the users with new computer technologies for dealing with research and practical tasks related with examination of human HNA parameters in health and pathology. PMID- 15757085 TI - [Test computer systems in the diagnosis of cognitive disorders as observed in the syndrome of attention deficiency with hyperactivity in children]. AB - The instrumental methods of evaluation of the cognitive ability of child by means of test computer systems open up real opportunities for the objective and thorough examination of main processes of the cognitive ability. An objective quantitation of the cognitive sphere makes it possible to prognosticate the intellectual development of child and to use such methods not only in the diagnosis and prevention but also in the correction of cognitive disorders. PMID- 15757086 TI - [Scientific, organizational and methodological fundamentals for developing health saving technologies at high school and their medical-and-technical basis]. AB - The key issues related with maintenance and introduction of health-saving technologies into the educational process of a high school are demonstrated in the paper by the example of the "Integrated Program" implemented at Bauman Moscow State Technical University. Methods of preclinical screening and monitoring of students' health including organizational principles, information technologies and psychophysical examination are described. PMID- 15757087 TI - [Specificity of high psychic functions in children with impaired intellectual and speech development]. AB - Babies and preschoolers with impaired psychic and speech development underwent psychological and psychophysiological examinations. The purpose was to detect comparative peculiarities of psychic processes of the higher nervous activity and to evaluate objectively the cognitive and thought processes in children with different levels of intellectual development. The results can be used in the differential diagnosis of impairments of the psychic and intellectual development as well as in the detection of individual specific features of a child while defining a scope of medical, psychological and pedagogical aid. PMID- 15757088 TI - [Game biofeedback: networking]. AB - Analyzed in the paper are stages of networking of game biofeedback. Three main stages are defined on the basis of practical experience, they are: unification of local databases, use of game biofeedback in local network and training in game biofeedback via Internet. Possible solutions are suggested; technical and organizational issues are discussed on working out and using a set of therapeutic and health-improving training equipment based on the principles of LAN game biofeedback. PMID- 15757089 TI - [Instrumental implementation of monitoring methods applicable to generation, tracing, reproduction and prognostication in deterministic and stochastic media]. AB - The case study is based on the theory of functional systems described by Academician P.K. Anokhin and on an experimental model of space-time prognostication enabling the development and classification of test-procedures modality of an activity (generation, tracing, reproduction and prognostication) and by modality (dimension) of space (zero-, 1- and 2D etc.). Test procedures were suggested and investigated for evaluating the parameters of the central nervous system (CNS). A set of experimental test models can be presented as a combination of described space-time modes. The system of indices is based on an evaluation of the time structure of a behavioral model. The activity rate parameter and its variability (mean value and root-mean-square deviation of the trend) were made use of. The rate of erroneous responses in different pattern's points was used as an evaluation tool for the spacious structure of the behavior. PMID- 15757091 TI - [Home computer stabilography: technical level, functional potentialities and spheres of application]. AB - Described, compared and analyzed in the paper are data about sabilographic computer equipment manufactured serially by the leading foreign and Russian companies. Potential spheres of application of stabilographic equipment are discussed. PMID- 15757090 TI - [Mechanisms of psychophysiological choice in schoolchildren and humanitarian students]. AB - Described in the paper is a case study targeted at detecting the age-related dynamics of some indices and psychophysiological mechanisms of decision-making in senior schoolchildren and humanitarian students; the study was based on computer aided test systems. The research methods are presented and its results are analyzed. PMID- 15757093 TI - [Potentialities of the dynamic tracing of the integrated activity of the brain]. AB - The paper deals with two aspects of psychophysiological examination made within the psychophysiological evaluation of the condition of healthy persons as well as within the psychophisiological express-diagnosis of latent and initial stress reactions. The level of psychophysiological indices ensuring the integrated functioning of the CNS was evaluated by the KPFK-99 "Psyhomat" automated set designed for psychophysiological diagnosis of man. Besides, its regulatory components were evaluated by means of the "VNS-spectrum" equipment: VNS parameters were analyzed (rhythmological evaluation of the vegetative tonus, vegetative reactivity and vegetative support). The psychological status was examined by a set of psychodiagnostic and psychosocial tests. PMID- 15757092 TI - [Examination of the equilibrium function in children aged 7 to 10 by means of "stabilotest" equipment]. AB - The paper contains the results of a study of the equilibrium function examined in 120 schoolchildren (aged 7 to 8 and 9 to 10) residing in the city of Arkhangelsk by means of a computer-based "Stabilotest" set manufactured by "VNIIMP-VITA", Moscow. Quantitative parameters of the functions were obtained for healthy schoolchildren of the above age groups on the basis of a variety of tests, which exposed a number of specific features related with maintaining the vertical postures in different positions. The results can be used as a basis in evaluating the functional status of the equilibrium system during diagnostic and correction and-rehabilitation procedures. PMID- 15757094 TI - [The psychoemotional condition of children in endoscopic examination]. AB - Described in the paper is the instrumental examination of the psychoemotional condition of schoolchildren with esogastroenterologic pathology, who underwent, for the diagnostic purpose, esogastroduodenoscopy. It is demonstrated that the determination of the psychoemotional status in the course of dynamic follow-up optimizes the preparation of a child for endoscopy and smooths the anxiety during the procedure. PMID- 15757095 TI - [Examination of the energetic condition of the brain in children and teenagers by means of "Neuroergometer-03"]. AB - Presented are the results of examination of the functional condition of the brain in 347 boys aged 5 to 16 residing permanently in the city of Arkhangelsk. High energy usage by the brain was registered in northerner children and teenagers versus their coevals residing in Russia's central regions. The principle of "cupola-shaping" in the distribution of the brain's permanent potential was found to be impaired. Besides, the cerebral hemisphere asymmetry was also affected in the northerners. The mentioned lesions denote the functional CNS tension in children and teenagers residing permanently in the European North. The method of registration of the level of the brain's constant potential enables an objective evaluation of the body's functional reserve. PMID- 15757096 TI - [Diagnosis of athletic abilities in students by psychophysiological equipment]. AB - Represented are issues related with individual prognostication of an athletic specialization made on the basis of diagnosing the athletic abilities of a person by means of "Psihomat-99" automated set designed for psychophysiological monitoring. Examination methods as well as results and conclusions are described. PMID- 15757097 TI - Residency program director and medical center liable for withdrawal of recommendation. Driscoll v. Stucker. PMID- 15757098 TI - Eighth Circuit rules on punitive damages award. Stogsdill v. Healthmark Partners. PMID- 15757099 TI - Shortness of patience: does serving always mean transporting? PMID- 15757101 TI - Highway safety focus of vehicle safety initiative. PMID- 15757100 TI - Easy as P.I.E. PMID- 15757102 TI - Can we leave the scene? PMID- 15757103 TI - A deadly season: hurricanes tax EMS providers in Florida. PMID- 15757104 TI - Car bomb response. PMID- 15757105 TI - Lessons from Chernobyl. PMID- 15757106 TI - Recognizing imminent danger: characteristics of a suicide bomber. AB - By all reports, the public is at increasing risk of attack by terrorist organizations. Citizens have been asked to increase their vigilance and report suspicious individuals to reduce the chance of a suicide or homicide bombing. Terrorist groups often target their actions to inflict the most casualties and may choose mass gatherings to detonate an explosive device. Since EMS providers are frequently assigned to work special details where large numbers of people may gather, EMTs and paramedics can assist law enforcement in identifying individuals who may pose a threat to the people gathered at the event. The characteristics of suicide bombers discussed here are meant to increase your awareness level to recognize a potential problem. The astute EMS provider can then notify appropriate law enforcement personnel for further investigation and action. PMID- 15757108 TI - Performance-enhancing drugs and supplements. PMID- 15757107 TI - Are we ready for suicide bombings? PMID- 15757109 TI - Heart Healthy Firefighter Program. PMID- 15757110 TI - EMS for the vertically challenged. PMID- 15757111 TI - Do you have the courage to change? PMID- 15757112 TI - Fast and good: trimming fast food to suit your size. PMID- 15757113 TI - Gym on a shoestring. PMID- 15757114 TI - Influence of normovolemic anemia on Doppler characteristics of the abdominal aorta and splanchnic vessels in Beagles. AB - OBJECTIVE: To ultrasonographically evaluate hemodynamics in the abdominal aorta (AAo) and splanchnic vessels in dogs with experimentally induced normovolemic anemia. ANIMALS: 11 healthy Beagles. PROCEDURE: The AAo, cranial mesenteric artery (CMA), celiac artery (CA), hilar splenic artery (HSA), and main portal vein (MPV) were evaluated in conscious dogs immediately before and after experimental induction of severe normovolemic anemia (Hct, 16%) and during recovery from moderate and mild anemia (Hct, 26% and 34%, respectively). Peak systolic velocity (PSV) or peak velocity (PV), time-averaged mean velocity (TAVmean), pulsatility index (PI), resistive index (RI), blood flow, congestion index (CI), and heart rate (HR) were recorded. Results were compared for anemic and control states. RESULTS: Severe anemia caused significant increases in HR (25% to 70%), PSV (AAo, 45.8%; CMA, 56.1%; and CA, 41.9%), PV (MPV, 84.2%), and TAVmean, (AAo, 69.4%, CMA, 64.3%; CA, 29.7%; and MPV, 76.9%) and significant decreases in PI (AAo, 26.1%; HSA, 19.3%) and Cl (MPV, 45.2%). There was no significant change in PI of the CMA or CA, portal blood flow, or RI of any artery. Significantly higherTAVmean persisted in all vessels during moderate anemia, but higher PSV persisted only in the CMA; PI (CMA and CA) and RI (CA) decreased significantly, but portal blood flow increased significantly. Significant increase in TAVmean (AAo and CMA) persisted during mild anemia, and PI (AAo, CMA, and HSA) and RI (CMA) were significantly lower. CONCLUSIONS AND CLINICAL RELEVANCE: Doppler ultrasonography revealed hyperdynamic circulation in the AAo and splanchnic vessels in dogs with experimentally induced normovolemic anemia. PMID- 15757116 TI - Culture of feline corneal epithelial cells and infection with feline herpesvirus 1 as an investigative tool. AB - OBJECTIVE: To isolate and characterize pure cultures of feline corneal epithelial cells and to assess the extent and nature of feline herpesvirus (FHV)-1 infection in these cells. SAMPLE POPULATION: Healthy eyes from 23 recently euthanatized cats. PROCEDURE: Stroma and epithelium of the rostral portion of the cornea were surgically isolated, and epithelial cells were detached from the stroma by enzymatic incubation. Epithelial cells were cultured in hormone-supplemented media. Cells were passaged, and cytokeratin expression was assessed. Cells were then infected with FHV-1, and cytopathic effects were determined. RESULTS: Cell cultures were readily established from samples obtained from each eye and could be maintained through 6 passages. Cultured cells expressed cytokeratins 3 and 12 but not other cytokeratins. Infection with FHV-1 was rapid and caused widespread cytopathic effects. CONCLUSIONS AND CLINICAL RELEVANCE: Feline corneal cells cultured in vitro during multiple passages maintain consistent morphologic characteristics and intermediate filament expression. They are susceptible to infection with FHV-1 and may provide a useful in vitro model for investigation of ocular drugs. PMID- 15757115 TI - Effects of dietary antioxidant supplementation before and after oral acetaminophen challenge in cats. AB - OBJECTIVE: To determine effects of lipoic acid, vitamin E, and cysteine before and after oxidant challenge in cats. ANIMALS: 24 sexually intact adult cats. PROCEDURE: Cats were allocated into 4 equal groups. For 25 weeks, group A was fed a control dry diet and groups B, C, and D received this diet supplemented with vitamin E (2200 U/kg [dry matter basis {DMB}]) plus cysteine (9.5 g/kg [DMB]), lipoate (150 mg/kg [DMB]), or all 3 antioxidants together, respectively. Weights were measured every 3 days and venous blood obtained every 5 weeks for CBC; serum biochemical analyses; lymphocyte blastogenesis; thiobarbituric acid reactive substances concentration; and concentrations of plasma protein carbonyl, 8-OH d guanosine, blood glutathione, plasma amino acid, lipoate, and dihydrolipoate. At 15 weeks, all cats received acetaminophen (9 mg/kg, PO, once), clinical effects were observed, and methemoglobin concentrations were measured. RESULTS: Lymphocyte blastogenesis increased transiently in group C and D cats. After acetaminophen administration, all groups had transient increases in methemoglobin within 4 hours and mild, brief facial edema; group C had decreased glutathione concentration and increased 8-OH d-guanosine concentration versus controls; and protein carbonyl concentration increased least for group B. Plasma lipoate and dihydrolipoate concentrations peaked by week 10 for groups C and D. Conclusions and Clinical Relevance-Lipoate, vitamin E, and cysteine did not have synergistic effects. Lipoate supplementation (150 mg/kg [DMB]) did not act as an antioxidant but appeared to enhance oxidant effects of acetaminophen. Vitamin E plus cysteine had protective effects. PMID- 15757117 TI - Effects of interferon-alpha on cytopathic changes and titers for feline herpesvirus-1 in primary cultures of feline corneal epithelial cells. AB - OBJECTIVE: To assess the effect of interferon (IFN)-alpha on viability of feline corneal epithelial cells, replication of feline herpesvirus (FHV)-1, and virus induced cytopathic changes. SAMPLE POPULATION: Healthy eyes from 10 recently euthanatized cats. PROCEDURE: 4 replicate primary cultures of feline corneal epithelial cells were grown after the addition of 10(2) to 10(6) IU of IFN alpha/mL. Cultures were examined every 24 hours for evidence of cytotoxic changes. Viable cell counts and percentage of viable cells were determined 48 hours after initiation of culture. In a separate experiment, cultures of corneal cells were inoculated with FHV-1 and cultured for 72 hours with or without 10(5) IU of IFN-alpha/mL. The FHV-1-infected cultures were evaluated for viral-induced cytopathic effects, and viral titers were determined in samples of culture supernatant. RESULTS: Interferon-alpha did not have cytotoxic effects on corneal epithelial cells at concentrations ranging from 10(2) to 10(6) IU of IFN alpha/mL. Interferon-alpha at a concentration of 10(5) IU/mL significantly reduced the cytopathic changes and FHV-1 titers. CONCLUSIONS AND CLINICAL RELEVANCE: Lack of in vitro cytotoxic effects and efficacy against FHV-1 infection in primary cultures of feline corneal cells suggests that the in vivo therapeutic effect of IFN-a should be assessed in controlled clinical trials. PMID- 15757118 TI - Effects of cidofovir on cell death and replication of feline herpesvirus-1 in cultured feline corneal epithelial cells. AB - OBJECTIVE: To assess the effect of cidofovir on viability of feline corneal epithelial (FCE) cells, replication of feline herpesvirus (FHV)-1, and virus induced cytopathic changes. SAMPLE POPULATION: Healthy eyes from 14 recently euthanatized cats. PROCEDURE: Cidofovir at concentrations ranging from 0.05 to 0.000005 mg/mL was added to primary cultures of FCE cells, and cytopathic changes and effects on cell proliferation and cell viability were determined during the subsequent 48 hours. Efficacy of cidofovir (0.02 and 0.05 mg/mL) to prevent in vitro infection of FCE cells with FHV-1 was determined during 72 hours of culture by assessing viral cytopathic effects and viral titers. RESULTS: Cidofovir at concentrations of 0.05, 0.005, and 0.0005 mg/mL significantly reduced mean viable cell counts, and cidofovir at a concentration of 0.05 mg/mL significantly reduced the percentage viability of cultured FCE cells. Minimal cytopathic changes were observed at concentrations of 0.02 and 0.05 mg of cidofovir/mL. Cidofovir at concentrations of 0.05 and 0.02 mg/mL abrogated the cytopathic effects attributable to FHV-1 infection and reduced viral titers from > or =10(14) TCID(50)/mL to < or =10(3.5) TCID50/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Cidofovir in vitro was highly efficacious against FHV-1 infection of a primary culture of FCE cells but had cytostatic effects on cultured cells. PMID- 15757119 TI - Characterization of equine intestinal fatty acid binding protein and its use in managing horses with colic. AB - OBJECTIVE: To determine the nucleotide sequence of the equine intestinal fatty acid binding protein (I-FABP) gene, its expression in various regions of the gastrointestinal tract, and the use of measuring I-FABP in horses with colic. Animals-86 horses with colic. PROCEDURE: The mRNA sequence for the I-FABP gene was obtained by use of a rapid amplification of complementary DNA ends technique. Comparative I-FABP gene expression was quantitated by use of a real-time reverse transcription-polymerase chain reaction assay. Amounts of I-FABP in abdominal fluid and plasma were measured by use of an ELISA kit. Association between I-FABP concentrations and clinical variables was performed by nonparametric analysis, and associations of these variables with intestinal ischemia were determined by the Spearman correlation test. RESULTS: The nucleotide sequence had 87% identity with human I-FABP The I-FABP gene was highly expressed in the small intestinal mucosa but had low expression in the colon. High concentrations of I-FABP in abdominal fluid correlated with an increase in protein concentrations in peritoneal fluid and nonsurvival, whereas plasma I-FABP concentrations correlated with the necessity for abdominal surgery. Clinical variables associated with intestinal ischemia included the color and protein content of abdominal fluid and serum creatine kinase activity. CONCLUSIONS AND CLINICAL RELEVANCE: Determination of I-FABP concentrations in abdominal fluid and plasma may be useful for predicting survival and the need for abdominal surgical intervention in horses with colic. Furthermore, serum creatine kinase activity and color and protein concentrations of abdominal fluid may be useful in the diagnosis of intestinal ischemia. PMID- 15757120 TI - Evaluation of six-lead electrocardiograms obtained from dogs in a sitting position or sternal recumbency. AB - OBJECTIVE: To compare 6-lead ECG traces in clinically normal conscious dogs in a sitting position and sternal recumbency to that of right lateral recumbency. ANIMALS: 31 healthy dogs with no history of cardiac disease. PROCEDURE: Six-lead ECGs were recorded for dogs in right lateral recumbency, a sitting position, and sternal recumbency. Q-, R-, and S-wave amplitudes as well as QRS-complex duration were measured in all leads. Additionally, P-wave amplitude and duration, PR interval, ST-segment elevation or depression, and OT interval were measured in lead II. RESULTS: Compared with measurements in right lateral recumbency, the sitting position resulted in increased Q-wave amplitude (lead III), increased R wave amplitude (leads I and aVL), decreased R-wave amplitude (leads III and aVF), increased S-wave amplitude (lead aVR), decreased S-wave amplitude (lead aVL), increased P-wave amplitude (lead II), and a leftward shift in the mean electrical axis. Compared with measurements in right lateral recumbency, sternal recumbency resulted in decreased Q-wave amplitude (leads I, II, and aVF), increased R-wave amplitude (leads 11, III, and aVF), decreased R-wave amplitude (lead aVR), increased S-wave amplitude (lead aVR), increased P-wave amplitude (lead II), and decreased ST-segment depression (lead II). Compared with right lateral recumbency, the sitting position or sternal recumbency did not result in significant differences in PR interval, QT interval, or QRS-complex duration. CONCLUSIONS AND CLINICAL RELEVANCE: Significant changes are found in ECG measurements in the sitting position and sternal recumbency, compared with right lateral recumbency. In dogs, many ECG reference range values for right lateral recumbency are not valid for ECGs obtained in the sitting position or sternal recumbency. PMID- 15757121 TI - Plication of the free wall of the left ventricle in dogs with doxorubicin-induced cardiomyopathy. AB - OBJECTIVE: To evaluate plication of the free wall of the left ventricle, which reduces the left ventricular area and volume, as a method to improve the left ventricular systolic function without cardiopulmonary bypass. ANIMALS: 8 mixed breed adult dogs. PROCEDURE: Dilated cardiomyopathy (DCM) was induced in each dog by administration of doxorubicin (30 mg/m2, i.v., q 21 d for 168 days). Two dogs died during induction of cardiomyopathy. Plication surgery was performed in 4 dogs. Two dogs did not ondergo to surgery (control group). Values for cardiac output (CO), 2-dimensional and M-mode echocardiography, arterial blood pressure, electrocardiography, blood cell counts, and serum biochemical analyses were recorded after induction of DCM (baseline) and 1, 2, 7, 15, 21, 30, 60, 90, 120, 150, and 180 days after plication surgery. Ambulatory ECG (Holter) recordings were conducted for 24 hours on the day of surgery. RESULTS: 1 dog died after plication surgery. The remaining dogs undergoing ventricular plication had a significant improvement in CO, ejection fraction, and fractional shortening and reductions of left ventricular area and volume after surgery. Electrocardiographic and Holter recordings revealed premature ventricular complexes, which resolved without treatment during the first week after surgery. Clinical condition of the control dogs declined, and these 2 dogs died approximately 40 days after induction of cardiomyopathy. CONCLUSIONS AND CLINICAL RELEVANCE: Plication of the free wall of the left ventricle improved left ventricular systolic function in dogs with doxorubicin-induced cardiomyopathy. Additional studies are needed to evaluate its application in dogs with naturally developing DCM. PMID- 15757122 TI - Glucose, lactate, and pyruvate concentrations in dogs with babesiosis. AB - OBJECTIVE: To document changes in glucose, lactate, and pyruvate concentrations in dogs with severe or complicated babesiosis; assess relationships among glucose, lactate, and pyruvate concentrations in those dogs; and compare clinical and laboratory variables in dogs with and without hypoglycemia and hyperlactatemia. ANIMALS: 20 dogs with naturally developing severe or complicated babesiosis. PROCEDURE: Samples and measurements were obtained before treatment was initiated. Babesiosis was diagnosed by examination of blood smears. Arterial blood pressure measurement, parasite quantification, CBC count, serum biochemical analysis, urinalysis, venous blood gas analysis, and acid-base determination were performed. Glucose, lactate, and pyruvate concentrations were measured in samples of venous blood. RESULTS: We detected a significant negative correlation between glucose and lactate concentrations. Glucose, lactate, and pyruvate concentrations all differed significantly between dogs that died and those that survived. Three of 5 dogs that died had concurrent hypoglycemia, hyperlactatemia, and hyperpyruvatemia. Hypoglycemic dogs differed significantly from normoglycemic dogs with regard to lactate, urea, and bicarbonate concentrations; lactate-to pyruvate ratio; percentage parasitemia; and PCO2. Dogs with hyperlactatemia differed significantly from normolactatemic dogs with regard to clinical collapse; alanine transaminase activity; concentrations of bilirubin, urea, creatinine, and bicarbonate; percentage parasitemia; and PCO2. CONCLUSIONS AND CLINICAL RELEVANCE: Abnormal carbohydrate metabolism is commonly evident in dogs with severe or complicated babesiosis and is often associated with changes in other clinical and laboratory variables. Significant differences were found between survivors and nonsurvivors. Hypoglycemia should be assessed and aggressively treated in dogs with babesiosis. Lactate concentration can be used as an indicator of disease severity. PMID- 15757124 TI - Effects of sulfamethoxazole-trimethoprim on thyroid function in dogs. AB - OBJECTIVE: To evaluate effects of trimethoprim-sulfamethoxazole (T/SMX) on thyroid function in dogs. ANIMALS: 6 healthy euthyroid dogs. PROCEDURE: Dogs were administered T/SMX (14.1 to 16 mg/kg, PO, q 12 h) for 3 weeks. Blood was collected weekly for 6 weeks for determination of total thyroxine (TT4), free thyroxine (fT4), and canine thyroid-stimulating hormone (cTSH) concentrations. Schirmer tear tests were performed weekly. Blood was collected for CBC prior to antimicrobial treatment and at 3 and 6 weeks. RESULTS: 5 dogs had serum TT4 concentrations equal to or less than the lower reference limit, and 4 dogs had serum fT4 less than the lower reference limit after 3 weeks of T/SMX administration; cTSH concentrations were greater than the upper reference limit in 4 dogs. All dogs had TT4 and fT4 concentrations greater than the lower reference limit after T/SMX administration was discontinued for 1 week, and cTSH concentrations were less than reference range after T/SMX administration was discontinued for 2 weeks. Two dogs developed decreased tear production, which returned to normal after discontinuing administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of T/SMX at a dosage of 14.1 to 16 mg/kg, PO, every 12 hours for 3 weeks caused decreased TT4 and fT4 concentrations and increased cTSH concentration, conditions that would be compatible with a diagnosis of hypothyroidism. Therefore, dogs should not have thyroid function evaluated while receiving this dosage of T/SMX for >2 weeks. These results are in contrast to those of a previous study of trimethoprim-sulfadiazine. PMID- 15757123 TI - Morphologic and physiologic changes induced by Clostridium perfringens type A alpha toxin in the intestine of sheep. AB - OBJECTIVE: To evaluate the morphologic and physiologic changes induced by Clostridium perfringens type A (alpha toxin in the ileum and colon of sheep. SAMPLE POPULATION: 16 ligated intestinal loops in 4 Merino lambs and 18 explants of ileum and colon from slaughtered lambs. PROCEDURE: alpha Toxin-induced fluid accumulation was evaluated in ligated ileal and colonic loops of sheep. Tissues were evaluated morphologically by use of gross and histologic examination. Effects of toxin on in vitro intestinal net water transport were tested in modified Ussing chambers. RESULTS: Ovine ileal and colonic loops incubated with C perfringens type A alpha toxin retained more fluid than control loops. Histologically, in the ileum of lambs inoculated with 300 LD50 of alpha toxin/mL, there was a mild to moderate multifocal infiltration of neutrophils in the lamina propria and submucosa. The colonic loops of lambs inoculated with 30 or 300 LD50 of alpha toxin/mL had excessive mucus in the lumen, a moderate amount of neutrophils mixed with mucus in the intestinal lumen, and moderate multifocal infiltration of the lamina propria and submucosa with neutrophils; the blood vessels of these layers were engorged with neutrophils. In vitro measurements of water transport also revealed inhibition of net epithelial water absorption in ileum and colon incubated with alpha toxin on the mucosal side. CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that alpha toxin induces alterations in sheep intestine. Clostridium perfringens type A organisms that produce alpha toxin could be responsible for diseases of intestinal origin in some ruminants. PMID- 15757125 TI - Effects of exercise-induced stress and dexamethasone on plasma hormone and glucose concentrations and sedation in dogs treated with dexmedetomidine. AB - OBJECTIVE: To compare the effects of pretreatment with dexamethasone, physical stress (exercise), or both on sedation and plasma hormone and glucose concentrations in dogs treated with dexmedetomidine (DEX). ANIMALS: 6 healthy purpose-bred Beagles. PROCEDURE: Dogs received 4 treatments each in a randomized order prior to i.v. administration of DEX (5 fLg/kg). Pretreatments were as follows: (1) i.v. administration of saline (0.9% NaCI) solution and no exercise (control group); (2) IV administration of dexamethasone (0.05 mg/kg) and no exercise (DM group); (3) i.v. administration of saline solution and exercise (EX group; 15 minutes of trotting on a treadmill at a speed of 2 m/s); and (4) i.v. administration of dexamethasone and exercise (DM+EX group). RESULTS: Following DEX administration, all dogs had similar times to recumbency and sedation index values, irrespective of pretreatment with values, irrespective of pretreatment with dexam-d ethasone or exercise. Plasma catecholamine concentrations decreased after DEX administration. Compared with control group dogs, plasma cortisol concentrations were higher in EX-group dogs prior to DEX administration and lower in DM- and DM+EX-group dogs following DEX administration. Administration of DEX decreased plasma cortisol concentration in EX-group dogs only. Plasma glucose concentration was not influenced by exercise or dexamethasone administration was lower than baseline concentrations at 30 minutes after DEX administration and returned to baseline values by 90 minutes. Heart and respiratory rates and rectal temperature increased during exercise. After DEX administration, these values decreased below baseline values. The decrease in heart rate was of shorter duration in dogs that underwent pretreatment with dexamethasone, exercise, or both than in control group dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Pretreatment with dexamethasone, moderate physical stress (exercise), or both did not influence sedation or cause adverse effects in healthy dogs treated with DEX. PMID- 15757126 TI - Cloning and expression of type III collagen in normal and injured tendons of horses. AB - OBJECTIVE: To clone the 5' end of type III collagen and describe its pattern of mRNA and protein expression in normal and healing tendons in horses. ANIMALS: 14 healthy adult horses. PROCEDURE: The tensile region of collagenase-injured superficial digital flexor tendons was harvested at intervals from 1 to 24 weeks after injury. Total RNA was reverse-transcribed into cDNA for cloning and sequencing of type III collagen. Equine-specific nucleic acid probes were developed and used for northern blot analysis and in situ hybridization. Type III collagen protein and cyanogen bromide-cleaved collagen peptides were assessedby gel electrophresis. RESULTS: Type III collagen mRNA expression and protein content increased immediately after injury and remained increased. Type III collagen was localized to the endotenon in normal tendon and in injured tendon at 1 week. At 8 and 24 weeks, expression became more widely distributed throughout the tendon parenchyma. Injured tendon contained 6 times more type I than type III collagen mRNA. Quantities of type III collagen protein were maximal in the first 4 weeks after injury (approx 33%) and then began to decrease. CONCLUSIONS AND CLINICAL RELEVANCE: Type III collagen expression is increased initially in endotenon and subsequently in parenchyma of healing tendon; however, type III remains the minor collagen throughout the healing process. The role of type III collagen in tendon healing is not fully elucidated. PMID- 15757127 TI - Evaluation of glucose metabolism in three horses with lower motor neuron degeneration. AB - OBJECTIVES: To determine whether increased glucose metabolism is the potential cause of the decreased plasma glucose curve determined after oral glucose tolerance testing in horses with lower motor neuron degeneration. ANIMALS: 3 horses with signs suggestive of lower motor neuron degeneration, 1 horse with malignant melanoma with multiple metastases, and an obese but otherwise healthy horse. Procedures-Glucose metabolism was assessed by use of the hyperglycemic clamp and euglycemic hyperinsulinemic clamp techniques. RESULTS: Mean rate of glucose metabolism of horses with lower motor neuron degeneration was significantly greater (mean, 3.7 times greater than control horses; range, 2.1 to 4.8 times greater) than that reported in 5 healthy control horses (41 +/- 13 micromol/kg/min vs 11 +/- 4.5 micromol/kg/min, respectively). In addition, one of the affected horses, an 8-year-old warmblood gelding, had a 5.6-times increased sensitivity to exogenously administered insulin, compared with that reported in 5 healthy control horses. Pancreatic insulin secretion was not insufficient in horses with lower motor neuron degeneration. Findings in the 2 diseased control horses were unremarkable. CONCLUSIONS AND CLINICAL RELEVANCE: Increased glucose metabolism in horses with lower motor neuron degeneration may be the cause of the decreased plasma glucose curve detected after oral glucose tolerance testing. This finding could aid in developing supportive treatments with respect to adequate glucose and vitamin E supplementation. PMID- 15757128 TI - Evaluation of architectural changes along the proximal to distal regions of the dorsal laminar interface in the equine hoof. AB - OBJECTIVE: To describe architectural changes along the dorsal laminar interface of the equine foot. SAMPLE POPULATION: 6 macroscopically normal forefeet obtained from 6 equine cadavers. PROCEDURE: Histologic sections of 8 evenly spaced, proximal to distal, samples of the dorsal laminar interface were photographed, digitized, and examined for differences in architecture. Laminar depth; secondary laminar density; number and consistency of bifurcations occurring within the secondary laminae, and areas composed of primary dermal lamina, primary epidermal lamina, and secondary laminar interface were recorded. Data were examined to test for differences in architecture associated with the proximal to distal positioning of the sample. RESULTS: With exception of the areas of the primary epidermal and primary dermal lamina, all measured variables were significantly different between the proximal and distal regions of the dorsal laminar interface. Changes included increases in laminar depth and the secondary laminar density. Bifurcation of secondary laminae principally occurred proximally and had an increased depth of bifurcation distally. The secondary laminar dermal epidermal interface had a 109% increase in area between the most proximal and distal sections. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicate that the interface normally contributes a substantial volume of dermal components to the internal surface of the wall. These data also indicate that 2 distinct mechanisms (i.e., bifurcation of secondary laminae and an increase in the length of secondary laminae) contribute to changes in the architecture of the laminar interface, which allows for the hypothesis that the normal laminar interface is capable of responding to mechanical load. PMID- 15757129 TI - Use of force plate analysis to compare the analgesic effects of intravenous administration of phenylbutazone and flunixin meglumine in horses with navicular syndrome. AB - OBJECTIVE: To use force plate analysis to evaluate the analgesic efficacies of flunixin meglumine and phenylbutazone administered i.v. at typical clinical doses in horses with navicular syndrome. ANIMALS: 12 horses with navicular syndrome that were otherwise clinically normal. PROCEDURE: Horses received flunixin (1.1 mg/kg), phenylbutazone (4.4 mg/kg), or physiologic saline (0.9% NaCI; 1 mL/45 kg) solution administered IV once daily for 4 days with a 14-day washout period between treatments (3 treatments/horse). Before beginning treatment (baseline) and 6, 12, 24, and 30 hours after the fourth dose of each treatment, horses were evaluated by use of the American Association of Equine Practitioners lameness scoring system (half scores permitted) and peak vertical force of the forelimbs was measured via a force plate. RESULTS: At 6, 12, and 24 hours after the fourth treatment, subjective lameness evaluations and force plate data indicated significant improvement in lameness from baseline values in horses treated with flunixin or phenylbutazone, compared with control horses; at those time points, the assessed variables in flunixin- or phenylbutazone-treated horses were not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: In horses with navicular syndrome treated once daily for 4 days, typical clinical doses of flunixin and phenylbutazone resulted in similar significant improvement in lameness at 6, 12, and 24 hours after the final dose, compared with findings in horses treated with saline solution. The effect of flunixin or phenylbutazone was maintained for at least 24 hours. Flunixin meglumine and phenylbutazone appear to have similar analgesic effects in horses with navicular syndrome. PMID- 15757130 TI - Evaluation of cardiac lesions and risk factors associated with myocarditis and dilated cardiomyopathy in southern sea otters (Enhydra lutris nereis). AB - OBJECTIVE: To describe cardiac lesions and identify risk factors associated with myocarditis and dilated cardiomyopathy (DCM) in beach-cast southern sea otters. ANIMALS: Free-ranging southern sea otters. PROCEDURE: Sea otters were necropsied at the Marine Wildlife Veterinary Care and Research Center from 1998 through 2001. Microscopic and gross necropsy findings were used to classify sea otters as myocarditis or DCM case otters or control otters. Univariate, multivariate, and spatial analytical techniques were used to evaluate associations among myocarditis; DCM; common sea otter pathogens; and potential infectious, toxic, and nutritional causes. RESULTS: Clusters of sea otters with myocarditis and DCM were identified in the southern aspect of the sea otter range from May to November 2000. Risk factors for myocarditis included age, good body condition, and exposure to domoic acid and Sarcocystis neurona. Myocarditis associated with domoic acid occurred predominantly in the southern part of the range, whereas myocarditis associated with S. neurona occurred in the northern part of the range. Age and suspected previous exposure to domoic acid were identified as major risk factors for DCM. A sample of otters with DCM had significantly lower concentrations of myocardial L-carnitine than control and myocarditis case otters. CONCLUSIONS AND CLINICAL RELEVANCE: Cardiac disease is an important cause of death in southern sea otters. Domoic acid toxicosis and infection with S. neurona are likely to be 2 important causes of myocarditis in sea otters. Domoic acid-induced myocarditis appears to progress to DCM, and depletion of myocardial L-carnitine may play a key role in this pathogenesis. PMID- 15757131 TI - Cloning and expression of equine insulin-like growth factor binding proteins in normal equine tendon. AB - OBJECTIVES: To define a portion of the nucleotide sequences of each of the 6 insulin-like growth factor (IGF) binding proteins (IGFBPs) in horses and describe patterns of messenger RNA (mRNA) and protein expression for IGFBPs in normal equine tendons. ANIMALS: 7 horses. PROCEDURE: Total RNA was extracted from the tensile region of normal superficial digital flexor tendons and reverse transcribed into complimentary DNA (cDNA). The cDNA was amplified via PCR, and products representing portions of each IGFBP were cloned and sequenced. Nucleotide sequences were used to deduce the amino acid sequences, and both nucleotide and predicted amino acid sequences were compared with those published for bovine, human, mouse, and ovine IGFBPs. Gene expression was quantitated by real-time PCR assay, and protein expression was evaluated by western ligand blot (WLB). RESULTS: Clones ranged in size from 262 to 522 bp and had high degrees of sequence homology with other mammalian species. Sequence homology was highest between bovine and equine IGFBPs (86% to 95%) and amongst the IGFBP-5 sequences from the various species (92% to 95%). Message for IGFBP-2 to -6, but not IGFBP 1, was expressed in normal tendon. Protein expression for IGFBP-2, -3, and -4 was detected byWLB in normal tendon and markedly increased in damaged tendons. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide basic information and tools needed for further characterization of the role of the IGF system in tendon healing and may lead to the ability to potentiate the response of healing tendon to exogenous IGF-I via concurrent manipulation of IGFBPs. PMID- 15757132 TI - Incidence, risk factors, and heritability estimates of hind limb lameness caused by hip dysplasia in a birth cohort of boxers. AB - OBJECTIVE: To determine incidence, risk factors, and heritability estimates of hind limb lameness caused by hip dysplasia in a birth cohort of Boxers. ANIMALS: 1733 Boxers from 325 litters. PROCEDURE: Status of Boxers with respect to clinical signs of canine hip dysplasia (cCHD) was registered during an 8-year period. Survival analysis accounted for dogs lost to follow-up. Effective heritability for developing cCHD was estimated by use of a proportional hazard model on the basis of the Weibull distribution. Parametric survival models were developed to identify the influence of potential risk factors. RESULTS: Cumulative hazard rate for cCHD from 7 weeks to 8 years of age was 8.5%. Dogs that were kept on a floor covered with a slippery material were 1.6 times as likely to develop cCHD, compared with dogs kept on a nonslippery floor. Risk of cCHD doubled in dogs from litters with a high preweaning mortality rate. Dogs that were neutered at 6 months prior to a diagnosis of CHD were 1.5 times as likely to develop cCHD, compared with sexually intact dogs. Dogs >5 years of age were 1.8 times as likely to develop cCHD, compared with younger dogs. Estimated effective heritability of cCHD was 0.11. In terms of the risk of cCHD in progeny, mean estimated breeding value (EBV) of the 10 best and 10 worst sires was -0.32 and 0.42, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Registration of Boxers that develop cCHD may provide a strategy for disease prevention. In addition to diagnostic evaluation of radiographs, sire EBVs provide useful information for breeding selection decisions. PMID- 15757133 TI - Laboratory measures of hemostasis and fibrinolysis after intravenous administration of epsilon-aminocaproic acid in clinically normal horses and ponies. AB - OBJECTIVE: To determine whether epsilon-aminocaproic acid (EACA) administered IV affects hemostasis and fibrinolysis in clinically normal horses and ponies. ANIMALS: 20 clinically normal adult horses and ponies. PROCEDURES: Blood samples were collected 24 hours before (baseline) and 1 and 5 hours after i.v. administration of a low dose (30 mg/kg) or high dose (100 mg/kg) of EACA. Platelet count, fibrinogen concentration, prothrombin time, partial thromboplastin time (PTT), D-dimer concentration, alpha2-antiplasmin activity, and thrombin-antithrombin complex concentration were measured. Values at 1 and 5 hours were compared with baseline values. RESULTS: hour after administration of a low dose of EACA, mean fibrinogen concentration was significantly lower than baseline concentration. Mean PTT was significantly shorter than the baseline value 5 hours after administration of a low dose of EACA. One hour after administration of 100 mg of EACA/kg, mean alpha2-antiplasmin activity was significantly higher than baseline activity. Mean fibrinogen concentration was significantly lower than baseline concentration 1 and 5 hours after administration of a high dose of EACA. Mean PTT was significantly shorter than the baseline value 5 hours after administration of a high dose of EACA. CONCLUSIONS AND CLINICAL RELEVANCE: i.v. administration of 30 and 100mg of EACA/kg to clinically normal horses significantly modified some laboratory measures of hemostasis, consistent with its known antifibrinolytic effects. Although enhanced clot maintenance and diminished bleeding were not directly assessed, the clinical use of EACA may benefit some patients. PMID- 15757134 TI - Effects of dietary supplementation with sodium chloride on urinary relative supersaturation with calcium oxalate in healthy dogs. AB - OBJECTIVE: To evaluate the effect of dietary supplementation with sodium chloride (NaCl) on urinary calcium excretion, urine calcium concentration, and urinary relative supersaturation (RSS) with calcium oxalate (CaOx). ANIMALS: 6 adult female healthy Beagles. PROCEDURE: By use of a crossover study design, a canned diet designed to decrease CaOx urolith recurrence with and without supplemental NaCl (i.e., 1.2% and 0.24% sodium on a dry-matter basis, respectively) was fed to dogs for 6 weeks. Every 14 days, 24-hour urine samples were collected. Concentrations of lithogenic substances and urine pH were used to calculate values of urinary RSS with CaOx. RESULTS: When dogs consumed a diet supplemented with NaCl, 24-hour urine volume and 24-hour urine calcium excretion increased. Dietary supplementation with NaCl was not associated with a change in urine calcium concentration. However, urine oxalate acid concentrations and values of urinary RSS with CaOx were significantly lower after feeding the NaCI supplemented diet for 28 days. CONCLUSIONS AND CLINICAL RELEVANCE: Dietary supplementation with NaCl in a urolith-prevention diet decreased the propensity for CaOx crystallization in the urine of healthy adult Beagles. However, until long-term studies evaluating the efficacy and safety of dietary supplementation with NaCl in dogs with CaOx urolithiasis are preformed, we suggest that dietary supplementation with NaCl be used cautiously. PMID- 15757135 TI - Evaluation of flow cytometric and automated methods for detection of activated platelets in dogs with inflammatory disease. AB - OBJECTIVE: To evaluate platelet surface-associated P-selectin, mean platelet component concentration (MPC), mean platelet component distribution width (MPCDW), mean platelet volume (MPV), and platelet distribution width (PDW) for detection of activated platelets in dogs with septic and nonseptic inflammatory disease. ANIMALS: 20 healthy dogs and 20 dogs with septic and nonseptic inflammatory disease. Procedures-Platelet surface-associated P-selectin (expressed as the median fluorescence intensity [MFI] of the platelet population), MPC, MPCDW, MPV, and PDW were determined in 20 healthy adult dogs, and reference ranges were calculated. These parameters were also determined in 11 dogs with nonseptic and 9 dogs with septic inflammatory disease and evaluated to determine which parameters were useful for detection of activated platelets. Results-12 dogs with inflammatory disease had P-selectin greater than the upper limit of the reference range, whereas 16 dogs with inflammatory disease had MPC lower than the lower limit of the reference range. All dogs in which P-selectin was greater than the upper limit of the reference range had MPC lower than the lower limit of the reference range. The correlation coefficient for P-selectin and MPC was 0.62. Differences in the MPCDW, MPV, and PDW in most dogs with inflammatory disease (compared with healthy dogs) were found; however, the correlation coefficients for P-selectin and MPCDW, MPV, and PDW were low. CONCLUSIONS AND CLINICAL RELEVANCE: Platelet surface-associated P-selectin and MPC appeared to be useful to detect activated platelets in most dogs with septic and nonseptic inflammatory disease. PMID- 15757136 TI - Evaluation of the influence of S-adenosylmethionine on systemic and hepatic effects of prednisolone in dogs. AB - OBJECTIVE: To evaluate the influence of a 1,4-butanedisulfonate stable salt of S adenosylmethionine (SAMe) administered orally on clinicopathologic and hepatic effects induced by long-term administration of prednisolone in dogs. ANIMALS: 12 healthy dogs. PROCEDURE: Following a pilot study (4 dogs), 2 groups of 4 dogs received prednisolone (2.2 mg/kg) orally once daily (84-day trial). One group received SAMe (20 mg/kg/d divided in 2 doses) for 42 days and then a placebo for 42 days; the other group received treatments in the reverse order. Before and during the trial, numerous variables were monitored, including serum total alkaline phosphatase (ALP) and glucocorticoid-induced ALP (G-ALP) activities, serum haptoglobin concentration, and total and oxidized glutathione (TGSH and GSSG) and thiobarbiturate-reacting substances (TBARS) concentrations in erythrocytes and liver tissue (days 0, 42, and 84). Hepatic specimens also were examined microscopically. RESULTS: The stable salt of SAMe was biologically available; plasma concentrations of SAMe or prednisolone were not affected by coadministration. Compared with baseline values, serum ALP and G-ALP activities and haptoglobin concentrations increased and erythrocyte GSSG and TBARS concentrations decreased with both treatments. Erythrocyte TGSH concentration decreased with the prednisolone-placebo treatment. Administration of SAMe appeared to conserve erythrocyte TGSH values and did not inhibit hepatocyte glycogen vacuolation but increased hepatic TGSH concentration and improved the hepatic tissue GSSG:TGSH ratio. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, administration of 20 mg of SAMe/kg/d may mitigate the apparent pro-oxidant influences of prednisolone but did not block development of classic clinicopathologic or histologic features of vacuolar hepatopathy. PMID- 15757138 TI - Women in the military. PMID- 15757139 TI - [Minimally invasive surgery in gastroenterology]. PMID- 15757137 TI - Comparison of pharmacokinetics of danofloxacin and enrofloxacin in calves challenged with Mannheimia haemolytica. AB - OBJECTIVE: To compare concentrations of danofloxacin, enrofloxacin, and ciprofloxacin in plasma and respiratory tissues of calves treated after challenge with Mannheimia haemolytica. ANIMALS: 75 calves. PROCEDURE: 24 hours after challenge with M. haemolytica, 72 calves with clinical signs of respiratory tract disease were randomly assigned to 1 of 12 equal treatment groups. Three nonchallenged, nontreated calves formed a control group. Challenged calves were treated with danofloxacin (6 and 8 mg/kg, SC) and enrofloxacin (8 mg/kg, SC) once. At 1, 2, 6, and 12 hours after treatment, 6 calves from each treatment group were euthanatized. Antimicrobial drug concentrations were assayed in various specimens. Peak plasma concentration (Cmax)-to-minimum inhibitory concentration (MIC; Cmax-to-MIC) ratios and the area under the concentration versus time curve over a 12-hour period-to-MIC ratios (AUC(12h)-to-MIC) were calculat-ed. RESULTS: Danofloxacin and enrofloxacin had MICs of 0.03 microg/mL for the M. haemolytica challenge isolate. Danofloxacin administered at doses of 6 and 8 mg/kg resulted in numerically higher geometric mean concentrations of danofloxacin in plasma and all respiratory tissues than geometric mean concentrations of enrofloxacin after treatment with enrofloxacin. Geometric mean concentrations of enrofloxacin were numerically higher than geometric mean concentrations of ciprofloxacin metabolite in plasma and almost all respiratory tissues. Danofloxacin and enrofloxacin achieved Cmax-to-MIC ratios >10 and AUC(12h)-to-MIC ratios >125 hours. CONCLUSIONS AND CLINICAL RELEVANCE: When used to treat pneumonic pasteurellosis in calves, danofloxacin and enrofloxacin can be expected to deliver concentration-dependent bactericidal activity against M. haemolytica, the bacteria most commonly associated with bovine respiratory tract disease. PMID- 15757140 TI - Laparoscopic management of idiopathic esophageal achalasia. AB - INTRODUCTION: Idiopathic esophageal achalasia was described in 1682 as a motor disorder of smooth muscle. In 1913 transabdominal cardiomyotomy was introduced and in 1958 this technique was popularized via a left thoracotomy, in 1991 laparoscopic cardiomyotomy was introduced since then many surgeons have adopted this technique as the primary treatment of esophageal achalasia. OBJECTIVE: Evaluate usefulness and recent advances in endoscopic surgery for the management of esophageal achalasia, as well as general issues in clinical presentation and diagnosis. METHODS: We did a review of the medical literature and of our series of patients treated between 1996 and 2004. RESULTS: Between 1996 and 2004, 84 patients underwent laparoscopic Heller myotomy and partial funduplication (78 Toupet and 6 Dor) for the treatment of achalasia. Operative time ranged from 2 to 3 hours with a mean of 2.5 hours. Mucosal perforation (esophageal/gastric) occurred in 3 patients and was repaired with primary closure. During follow-up 3 patients required postoperative endoscopic dilation and one required reoperation due to adhesions causing angulation of the g-e-junction. CONCLUSIONS: Laparoscopic Heller myotomy provided marked reduction in dysphagia, with minimal complaints of regurgitation, heartburn and chest pain at a mean follow-up of 2 years. PMID- 15757141 TI - [Cardias mucosa plication with endoluminal sutures by endoscopy in gastroesophageal reflux disease treatment]. AB - INTRODUCTION: Gastro-esophageal reflux disease is very common among general population, today's standard treatments are medical with proton pump inhibitors and laparoscopic antireflux surgery. Endoscopic gastric plication has been described recently for the treatment of this disease. OBJECTIVE: Review recent advances of the endoscopic gastric plication procedures for the management of gastro-esophageal reflux disease and report our experience. METHODS: Patients with symptomatic gastro-esophageal reflux disease are selected, with esophagitis A and B according to Los Angeles classification and with a De Meester score higher than 26 in 24 hour pH determination. Patients younger than 18 or older than 60 years old are excluded as well as patients with a hiatal hernia greater than 3 cm, previous esophageal or gastric surgery, associated esophageal pathology or a C or D esophagitis. RESULTS: After this procedure the lower esophageal sphincter increases its length, nevertheless there are minimal changes in its pressure, it also diminishes esophageal acid exposure time. Most studies show improvement in symptoms and less drug requirements. In studies with greater follow up period only a minority of patients persists with a good complete response with this treatment. CONCLUSIONS: Prospective randomized trials are needed to determine the real utility of this procedure, aside from these protocols there are no true indications to do this procedure in the general population. PMID- 15757142 TI - [Gastroesophageal reflux disease: Candidates and indications to surgical treatment]. AB - INTRODUCTION: Always, when the gastric juice gets into the esophagus, it produces changes in it. What brings the attention in this fact is that there can be no changes in the mucosa in spite of the reflux. The lost of co-relation with the clinic symptoms have created confusion in the patients evaluations. The gastroesophageal reflux disease is the result of a multifactor process in which esophagus and stomach are involved. OBJECTIVE: To know the indications for the surgical treatment. BACKGROUND: What has been the evolution on the management of the gastroesophageal reflux disease? In the first half of the twentieth century, the treatment was medical. In the fifty's, R. Nissen designed the anti-reflux surgery. The introduction of the proton pump inhibitors substantially improved the illness control with medications. The laparoscopic surgery appears, and the short time of recovery increased the interest on this therapeutic way. The availability of the ambulatory pH monitoring improved the possibility to recognize gastroesophageal reflux disease. The medical treatment has the advantage of been conservative and it can be lead to modify the cause and the effect. The surgical treatment can correct the anatomic defects, but does not resolve the acid secretion in most of the surgical techniques. In conclusion, there is no evidence to advocate medical or surgical therapy as the best therapy for gastroesophageal disease. As everything in medicine, the final decision on the gastroesophageal reflux disease must be individualized. PMID- 15757143 TI - [Current status and long-term results of laparoscopic anti-reflux surgery]. AB - BACKGROUND: Laparoscopic surgery has a great application for the treatment of different pathologies of the gastrointestinal tract. In the management of gastroesophageal reflux disease it is not a exception, since its introduction in 1991 this technique has evolved constantly. OBJECTIVE: Analyze and evaluate indications, surgical technique and long term results of laparoscopic surgery as a treatment for gastro-esophageal reflux disease. PATIENTS AND METHODS: In this review paper we analyze literature to evaluate the current status of laparoscopic surgery and its application to the treatment of gastro-esophageal reflux disease. Besides we briefly report long term results from authors experience. RESULTS: Indications for laparoscopic treatment of patients with gastroesophageal reflux disease include patients with moderate to severe erosive esophagitis, patients with non-typical symptoms in which a 24 hours pH measurement shows these symptoms are correlated with gastroesophageal reflux and those who do not tolerate medical treatment, also Barrett's disease is considered a relative indication. Laparoscopic surgery has increased the number of patients treated surgically because it is well tolerated. Long term results in terms of controlling gastroesophageal reflux must be superior to 90% with a low complication rate. CONCLUSIONS: Laparoscopic surgery for the treatment of gastroesophageal reflux disease has shown better recovery, hospital stay, return to daily activities and global morbidity, which our group has been able to reproduce. Selection criteria must be strict with an adequate long term follow up. PMID- 15757144 TI - [Actual status of laparoscopic cholecystectomy]. AB - INTRODUCTION: Since the first laparoscopic cholecystectomy in 1988, the management of gall-bladder disease has changed importantly. This technique was rapidly popularized in the U.S. as well as in Europe. Multiple studies have proved its feasibility, safeness and great advantages. OBJECTIVE: Analyze usefulness and recent advances of endoscopic surgery in the management of gallbladder disease. METHODS: We did a review of the recent medical literature to determine the actual status of laparoscopic cholecystectomy. RESULTS: Laparoscopic cholecystectomy is the most common surgical procedure performed in the digestive tract. During the year 2001, 1,100,000 cholecystectomies were done in the U.S., 85% were done laparoscopically. In Mexico cholecystectomy in government hospitals is done laparoscopically in 50% of the cases, while in private hospitals it reaches 90%. There are multiple prospective controlled studies showing superiority of laparoscopic cholecystectomy in times of recovery, costs, return to normal activity, pain, morbidity, esthetics among other advantages. CONCLUSIONS: Laparoscopic cholecystectomy is the gold standard for the treatment of the great majority of cases of gallbladder disease, nevertheless in developing countries open cholecystectomy is still done frequently. PMID- 15757145 TI - [Bile duct laparoscopic exploration]. AB - The use of ERCP with endoscopic esphincterotomy (ES) for the management of choledocholithiasis has replaced almost completely common bile duct exploration in the era of laparoscopic cholecystectomy. Once the procedure is completed it is followed by laparoscopic cholecystectomy in the same hospitalization. During the last few years, the development of new technology and equipment with possibilities of minimal invasive procedures and diagnosis, as well as the ongoing surgical skills has allowed surgeons to solve the problem of choledocholithiasis in one minimal invasive procedure, decreasing effectively morbidity with a high success rate, changing again ERCP with ES as a complement and not as a substitute for surgery. Nevertheless the initial step in the use of these techniques require of a special technical support and what is most important capacitation to achieve the desire objectives. Transcystic technique seems to be the most promising and choledochotomy with primary closure in cases that cannot be solved with the transcystic approach. Regardless of the approach the surgeon must be familiar with both and increasingly with the use of the endoscope in order to provide patients with advantages of minimal invasive surgery and all its advantages. We also discuss our series of 81 patients operated on with laparoscopic surgery with a high rate of success, low morbidity and mortality. PMID- 15757146 TI - [Endoscopic surgery in neoplasms of the upper digestive tract]. AB - INTRODUCTION: Endoscopic surgery has been performed since 1987 when in France the first laparoscopic cholecystectomy was done. Since then it has evolved and found a place in surgical practice. In the field of surgical oncology it has allowed the diagnosis, staging, resection and palliation of a great variety of neoplasms. OBJECTIVE: evaluate usefulness and recent advances of endoscopic surgery in neoplasms of the upper digestive tract, applied specifically to the esophagus and stomach. METHODS: We did a review of the medical literature using Medline data base (Pubmed). All series of cases, cohort and controlled studies published between January 1995 and April 2003 were included. RESULTS: For esophageal carcinoma as well as for gastric carcinoma, endoscopic surgery is used as a complement for computed tomography and endoscopic ultrasound to define clinical stage, increasing sensibility to detect metastatic disease. Techniques of endoscopic or video assisted esophageal transhiatal and thoracoscopic resection have been described, as well as Japanese studies including radical gastric resections in early gastric cancer with multiple advantages. There are also several developing possibilities of palliative endoscopic or video assisted treatment which are discussed in this article. CONCLUSIONS: Usefulness of endoscopic surgery in staging gastric and esophageal carcinoma is proven. Nevertheless oncologic endoscopic resections are still experimental and the advantages between these and open surgery are still to be shown with prospective randomized trials. PMID- 15757147 TI - [Minimally invasive surgery of small intestine]. AB - Since the first laparoscopic surgery was performed by Erick Muhe in 1986, multiple minimally invasive techniques have been developed. The small intestine presents several technical difficulties for the application of these techniques, namely: its size, mobilization inside the abdominal cavity, difficulty defining the site under manipulation and the distension association by an ileo or an occlusion. The development of new technology with better cameras and coagulation equipment such as the Harmonic Scalpel and the Ligasure, cutting linear staplers, have permitted the evolution of surgical techniques for the small intestine. Indications for the use of minimally invasive interventions in the treatment of the small intestine are classified as urgent and elective. Lesions secondary to an abdominal cavity puncture were the first to be treated in the small intestine. Abdominal trauma due to a sharp instrument can also be treated through this method. Intestinal occlusion treated without a great dilation can be solved in 55 to 65% of cases. Crohn's disease various surgical treatments are performed by laparoscopy. Obesity surgical treatments, particularly derivative gastroplasty have been carried out successfully by minimally invasive surgery. Gastrojejunoanastomosis or cystojejunoanastomosis are specialized techniques which are performed with minimal invasion; any kind of digestive or urinary stoma can be performed with these techniques. As the surgeon's surgical skills evolve, surgical minimally invasive indications for the small intestine become wider. PMID- 15757148 TI - [Appendectomy by minimally invasive surgery]. AB - OBJECTIVE: To analyze the results obtained with the laparoscopic appendectomy in a trial of 106 cases from 1997 to December of 2002 in a surgeons group. Comparing the results with the papers of the literature to qualified the safety and security of the method and to establish the advantages upon the open appendectomy. BACKGROUND DATA: The laparoscopic appendectomy is a technique that in the past ten years has proof its safety and efficacy to resolve the problems of vermiform appendix. METHODS: Prospective trial of 106 patients with probability or diagnosis of acute appendicitis approach to laparoscopic appendectomy. With statistical analysis of the data. RESULTS: 106 patients, 42 male and 62 female, average age 30.8, diagnosis of acute appendicitis before surgery of 70%, and with abdominal pain in 30%. Of the data obtained only the total count of leucocytes have the statistical value in the diagnosis of acute appendicitis. The average of surgical time was of 70.7 minutes, the average of length of hospital stay was of 74 hours. Diagnosis of acute appendicitis in 83% of the cases, normal appendix 10.3%, another pathology 6.7%. Conversion rate of 5.5% (6 patients). Abscess 3, peritonitis 1, hemorrhage 1, appendix rupture 1. Morbidity of 3.7% wound infection. Mortality 0%. CONCLUSIONS: The surgical indications for the laparoscopic appendectomy are the same that for open surgery. The technique is safe and effective that can be carried out in any patient, still in cases or perforated appendicitis or gangrenous. The success is depend of the experience of the surgical team. The wound infection, recovery time, postoperative pain are less in the laparoscopic appendectomy. PMID- 15757149 TI - Laparoscopic right hemicolectomy for cancer: 11-year experience. AB - INTRODUCTION: Laparoscopic surgery has emerged as the gold standard for many intra-abdominal procedures. Laparoscopic colon surgery is now entering its second decade of practice, and although there are many papers focusing on surgery of the distal colon, only a few have been published regarding right sided lesion approached totally laparoscopically. OBJECTIVE: Present data collected-in a prospective manner from a single institute over an eleven year period, focusing on laparoscopic right hemicolectomy for malignancy. METHODS: Patients elected for laparoscopic right hemicolectomy for colon cancer were analyzed prospectively. From May 1991 to May 2002, 98 patients underwent attempted laparoscopic right hemicolectomy for cancer, 44 male and 54 female, with a mean age of 70.6 years, emergent and non emergent cases were included Patients who underwent a diagnostic laparoscopy and those converted immediately to open procedure were excluded from this study. RESULTS: Ninety-two patients were included in the study, eighty-two of these had a totally intracorporeal anastomosis created, and ten had an extracorporeal anastomosis performed. The mean operative time for the intracorporeal group was 136 minutes, and for the extracorporeal group was 159 minutes. The average number of lymph nodes harvested was 10.8 and the final pathologic analysis showed 26 tumors stage I, 24 stage II, 31 stage III and 17 stage IV. CONCLUSIONS: In experienced hands, laparoscopic colectomy can be performed safely and effectively for the treatment of both benign and malignant diseases of the right colon. This study reaffirms the contention that laparoscopic approach to colon cancer offers equivalent, or in some instances, greater oncologic safety when compared to the open technique. PMID- 15757150 TI - [Total mesorectal excision by laparoscopy for rectal cancer]. AB - The treatment of rectal cancer has improved mostly because of the emergence of total mesorectal excision (TME), first described by Heald, and the adjunct of adjuvant therapy in the form of radiotherapy and chemotherapy. Laparoscopy has now been performed for more than a decade with results showing faster return to activity and decreased postoperative pain for intra-abdominal surgery. Laparoscopy is now widely used and is well accepted for the treatment of most intra-abdominal benign pathologies. Furthermore, several authors now report excellent short and medium-term results for the treatment of colorectal cancer. These study tends to show that laparoscopy can be safely applied to digestive malignancies, when oncological principles specific to laparoscopy are respected. The laparoscopic approach is in our opinion a perfect approach for the performance of TME, because of the magnification of structures that it provides, and allows the surgeon to see important structures that need to be identified for the performance of the best oncological and functional procedure possible. This paper describes in a through manner our standardized technique. We also review our own experience with laparoscopic TME, the multimodal treatment of rectal cancer, and provide an analysis of the literature about TME when performed by laparotomy and laparoscopy. PMID- 15757151 TI - [Laparoscopic surgery of pancreas]. AB - In pancreatic diseases, laparoscopic surgery is used to drain fluid collections in infected pancreatic necrosis, abscesses and pseudocysts, for cancer staging, to resect both benign and malignant tumors and for palliative procedures as well. In patients with intractable pain, percutaneous or surgical celiac plexus block can be performed, celiac ganglion alcoholization and torachoscopic splacnicectomy can be done. The laparoscopic approach for staging, allows us to identify very small peritoneal or hepatic metastasis, local invasion or deep hepatic metastasis. The lack of tactile sensation is a disadvantage in the laparoscopic procedures for both staging and resection; which can be compensated utilizing laparoscopic ultrasound. The laparoscopic pancreatic approach for resection is useful to treat neuroendocrine tumors, chronic pancreatitis and cystic neoplasm allowing preservation of the splenic vessels and the spleen. The insulinoma is a treatable laparoscopic tumor given its unique characteristics. The preoperative diagnosis is made by image studies like ultrasound, CAT scans, angiography studies, MRI, endoscopic ultrasound and calcium stimulation test. There are three surgical options: enucleation, distal pancreatectomy with or without preserving the spleen. The most important complication is the pancreatic fistula. CONCLUSIONS: The laparoscopic approach in pancreatic surgery is a feasible and safe one, although, its superiority over the classic open approach has not been demonstrated yet. PMID- 15757152 TI - [Laparoscopic liver surgery]. AB - INTRODUCTION: Laparoscopic surgery has been widely used for digestive tract diseases, specially for gallbladder pathology. Laparoscopic liver surgery pretends to accomplish the same objectives of open surgery but with the advantages of minimally invasive surgery. AIMS: To evaluate the indications and recent results of laparoscopic liver surgery, as well as the technique. MATERIAL AND METHODS: A review of the world literature was performed on the roll of laparoscopic surgery for hepatic diseases. The results of the authors are also reported. RESULTS: Nowadays hepatic laparoscopic surgery is reserved for anterior non-parasitic liver cysts and type I polycystic liver disease. The treatment of choice is wide deroofing. For hydatic liver disease pericystectomy or wide deroofing plus omentoplasty are also described. For benign solid liver disease, hepatic resection can be performed with the same outcome that the one observed with open surgery when the lesions are symptomatic, small, anterior or peripherally located. Laparoscopic liver surgery is recommended for the treatment of malignant hepatic lesions only in very specific cases. Nonetheless there are no prospective randomized studies that show benefits of laparoscopic over open surgery. Several local ablative therapies like radiofrequency, ethanol injection, cryotherapy or laser can be also performed through the laparoscope. The results reported so far have been encouraging. Intra-arterial chemotherapy is an option for the treatment of liver malignancies via laparoscopy, considering that the arterial catheter can be placed under this approach. CONCLUSIONS: For years, the liver has been considered a challenging organ for its approach, even with open surgery. Laparoscopic liver surgery should be performed by an experienced surgeon with the adequate technology. To our knowledge there are no reported results that decisively point toward the benefits of laparoscopic surgery over open surgery for the management of liver disease. Nonetheless more experience in laparoscopic liver surgery is needed. PMID- 15757153 TI - [Successful esophageal pH monitoring with Bravo capsule in patients with gastroesophageal reflux disease]. AB - INTRODUCTION: Esophageal 24-h pH monitoring (24-pH) is the most useful test to diagnose and treat patients with gastroesophageal reflux disease (GERD). The traditional system for 24-pH requires transnasal introduction of a catheter with pH sensors. This technique produces discomfort, inconvenience and interference with daily activity. Recently, the Bravo pH system has been proposed as an alternative and promising method for 24-pH. In this study, the initial experience in Mexico with this system is reported. OBJECTIVE: To evaluate safety, tolerability and performance of the pH Bravo capsule in patients with GERD. MATERIALS AND METHODS: Patients with GERD symptoms at least twice a week during the last three months, with indication for 24-pH were evaluated. pH Bravo capsule was placed 6 cm above squamocolumnar junction (SCJ). Symptoms, quality and duration of pH tracings, capsule detachment and patient global satisfaction were evaluated. RESULTS: Eleven patients (nine female, two male) mean age 42 years (range 26-62 years), two with erosive and nine with non-erosive GERD were studied. pH capsule was correctly positioned at 6 cm above SCJ in all patients. Nine patients noted a mild foreign body sensation (especially while eating) and four had mild chest pain; two patients had no discomfort. Capsule detachment occurred spontaneously in all patients on day 10. pH record for > 43 h was obtained in the 11 patients. There were no differences in pH parameters between days 1 and 2. Two patients with normal acid exposure on day 1 had abnormal pH parameters on day 2. CONCLUSIONS: Esophageal pH monitoring with Bravo capsule is a safe, reliable and tolerable method in patients with GERD. Extended pH recordings increases abnormal esophageal acid exposure detection in patients with this disease. PMID- 15757154 TI - [Efficacy, diagnostic utility and tolerance of intraesophageal pH ambulatory determination with wireless pH-testing monitoring system]. AB - BACKGROUND: Conventional catheter-based esophageal pH monitoring system produce discomfort and interference with normal activities and diet. An alternative is the wireless pH system which is more comfortable and offers extended recording periods. OBJECTIVE: To know the performance, the clinical utility and tolerability of wireless pH 48 h-monitoring system. PATIENTS AND METHODS: Forty one consecutive patients were randomized for conventional or wireless pH-testing. Successful placement of monitoring devices and data collection were evaluated. Frequency of pathologic gastroesophageal reflux was registred with both methods. A questionnaire was applied in all the cases to know the frequency of discomfort, interference with normal activities and diet related to the study. RESULTS: Conventional catheter-based pH-testing was performed in 21 patients and wireless system in 20. Successful placement of pH capsule was achieved in 20 of 22 patients and 48-h pH studies were completed in all of them. Pathologic gastroesophageal reflux was more frequently detected in the wireless system group (15 vs. 9 patients, p = NS). Ambulatory esophageal pH testing was well tolerated with both techniques. Chest discomfort was more frequently observed in the wireless group (11 vs. 2 patients, p = 0.005, RM 11.6, IC 95% 2.1-63.7). CONCLUSIONS: Ambulatory pH-monitoring wireless system is an efficient and well tolerated method which could detect a high proportion of subjects with pathologic reflux in comparison with conventional system. PMID- 15757155 TI - [Efficacy of two Helicobacter pylori eradication treatments in children with recurrent abdominal pain]. AB - OBJECTIVE: To compare clinical and bacteriologic efficacy of two therapeutic trials to eradicate Helicobacter pylori (H. pylori) in two series of pediatric patients with recurrent abdominal pain (RAP). MATERIALS AND METHODS: n = 36 children with RAP-associated H. pylori infection. Age 9.8 +/- 3.1 years, 19 boys and 17 girls. Clinical and bacteriologic efficacy of two therapeutic trials was compared: Group A (1996-1997), n = 9, amoxicillin, bismuth subsalicilate, and metronidazol, and group B (1991-1993), n = 27, omeprazol, amoxicillin, and clarithromycin. Initially and post-treatment, H. pylori evaluation was carried out with upper endoscopy and gastric biopsies. For statistics, we used Student t test, chi2, Fisher test, and Kruskal-Wallis analysis of variance (alpha = 0.05). RESULTS: We found that 33/36 cases had gastritis at endoscopy, two with duodenal ulcer; nodular gastritis was observed in more than one half of total cases. All cases fulfilled histologic criteria of gastritis according to Sydney Score. In group A eradication was achieved in 28.6%, while in group B eradication rose to 77.8% (p < 0.05). In group A, 8/9 and in group B 15/27 persisted with RAP (p = 0.113). CONCLUSIONS: High frequency of abnormal and histologic findings was observed in the series presented on children with RAP and H. pylori. Eradication efficacy in the omeprazol/amoxicillin/clarithromycin group was higher when compared with bismuth subsalicilate/amoxicillin/metronidazol trial. This efficacy is comparable to pediatric series treated with the same therapeutic trial. PMID- 15757156 TI - [Hemorrhoidal desarterialization guided by Doppler. A surgical alternative in hemorrhoidal disease management]. AB - BACKGROUND: Hemorrhoidal disease is a common condition that includes 30% of patients seen for the first time at the Colon and Rectal Unit of the Gastroenterology Service, Hospital General in Mexico City. This study shows experience with hemorrhoidal desarterialization guided by Doppler in patients with internal hemorrhoidal disease stages II or III in the Coloproctology Units of the Hospital General in Mexico City and at the North Medical Unit in Monterrey, Nuevo Leon, Mexico. MATERIALS AND METHODS: A prospective, longitudinal, and descriptive study was performed in 56 patients with diagnosis of internal hemorrhoids stages II or III, not complicated, and treated by hemorrhoidal artery ligation guided by Doppler between July 1, 2002 and April 31, 2003. RESULTS: 49 patients were included, 32 male (66%) and 17 female (34%), aged between 21 and 75 years with mean age of 43.5 years; 40 patients had internal hemorrhoids stage II (82%) and nine, stage III (18%). With a follow-up of 4 months or longer, all patients were symptomless: bleeding and hemorrhoidal prolapse disappeared. CONCLUSIONS: Doppler-Guided hemorrhoidal desarterialization is indicated in patients with internal hemorrhoids stages II or III; it is a simple technique that requires a short learning curve, minimal anesthetic and surgical materials, and pain after the procedure is mild and short-lived in the majority of patients. PMID- 15757157 TI - [Management of the abdominal wall in sepsis. Comparison of two techniques]. AB - OBJECTIVE: To compare two techniques of open abdomen management in patients with abdominal sepsis. INTRODUCTION: Some patients with abdominal sepsis will require open abdomen management to avoid abdomen compartment syndrome. We compare use of the Bogota bag with a technique developed at our Institution with a subcutaneous polyethylene bag. METHODS: Thirty nine consecutive patients with abdominal sepsis who were managed with open abdomen. RESULTS: Twenty one patients with Bogota bag and 18 patients with subcutaneous polyethylene bag. Patients with Bogota bag had more report surgery intrabdominal complications compared with patients with subcutaneous polyethylene bag (p = 0.04, p = 0.037, respectively). Mortality was the same. CONCLUSIONS: Use of subcutaneous polyethylene bag in patient with abdominal sepsis and open abdomen appears safer when compared with Bogota bag. PMID- 15757158 TI - [Cavernous hemangioma of the colon. Case report and review of the literature]. AB - Colon and rectum hemangioma is a rare disease, and even more so when it presents as cavernous subtype; few of these hemangioma have been confirmed by pathology reports. The main presentation of this disease is observed by transrectal painless bleeding; other manifestations are less frequent. Diagnosis implies high index of suspicion and proper diagnostic workup; this entity has been frequently misdiagnosed as other more common entities. We present the case of a 15-year-old female patient with a cavernous hemangioma of the sigmoid colon; her principal complaint was abdominal pain. Diagnosis was suspected due to the finding of phleboliths in plain abdominal film and diagnostic findings in colonoscopy. The patient was treated by means of segmental resection of the colon and experienced a satisfactory postoperatory recovery. According to the literature, these hemangiomas originate from embryologic sequestrations of mesodermal tissue, while clinically they usually present between the ages of 5 and 25 years, most commonly with painless transrectal bleeding and less frequently with intestinal obstruction, tenesmus, urgency, abdominal pain or mass, and anemia. The majority of these hemangiomas are localized in rectosigmoid region of the colon and may infiltrate surrounding tissues; 26-50% show phleboliths in plain abdominal film. Colonoscopy is considered by many authors the best diagnostic tool; angiography, computed tomography (CT) scan and scintigraphy may be also useful. The only effective treatment of this disease is complete surgical resection, for which there have been many proposals regarding best method and approach. PMID- 15757159 TI - [Osseous metaplasia in colonic adenocarcinoma]. AB - Osseous metaplasia (OM) in colonic adenocarcinoma is a very rare occurrence. This report describes the case of a 59-year-old man with OM occurring with moderately differentiated adenocarcinoma of colon, and is followed by a brief review of the literature. PMID- 15757160 TI - [Portal hypertension. Evidence-based guide]. AB - Treatment of portal hypertension has evolved widely during the last decades. Advances in physiopathology have allowed better application of therapeutic options and also have permitted to know the natural history of varices and variceal bleeding, predicting which patients have a higher risk of bleeding. It also permits probability of designing patient treatment. According to liver function and subadjacent liver disease, it is possible to offer different alternatives within the three possible scenarios (primary prophylaxis, acute bleeding episode, and secondary prophylaxis). For primary prophylaxis, pharmacotherapy offers the best choice. Endoscopic banding is also growing in these scenarios and probably will be accepted in the near future. For the acute bleeding episode, endoscopic therapy (sclerosis and/or bands) and/or pharmacologic therapy (octreotide, terlipresin) represent best choice, considering TIPS as a rescue option. Surgery is not used routinely in this scenario in most centers. For secondary prophylaxis, pharmaco- and endoscopic therapy are first-line treatments, while TIPS and surgery as second-line treatments. TIPS is mainly used in patients on a waiting list for liver transplantation. Surgery offers good results for low-risk patients, with good liver function and with portal blood-flow preserving procedures (selective shunts, extensive devascularizations). Liver transplantation is recommended for patients with poor liver function because together with portal hypertension, it treats subadjacent liver disease. PMID- 15757161 TI - [Polycystic liver disease]. PMID- 15757162 TI - [Diffuse esophageal spasm with atypical radiography]. PMID- 15757163 TI - [Ambulatory esophageal pH-monitoring with wireless system]. PMID- 15757164 TI - Trench foot, jungle rot, and now, Baghdad boil. PMID- 15757165 TI - Could cardiac CT revolutionize the practice of cardiology? PMID- 15757166 TI - Nonhealing skin lesions in a sailor and a journalist returning from Iraq. AB - US health care providers who are not familiar with cutaneous leishmaniasis may now begin to encounter more patients with this challenging entity as military personnel return from rotations in Iraq or Afghanistan. Diagnosis requires a skin scraping, aspiration, or biopsy, followed by examination by an experienced microscopist or pathologist. Demonstration of the parasite DNA by PCR or culture in special media can also be used to confirm the diagnosis. Sodium stibogluconate is the mainstay of therapy, but other options for selected cases include topical thermal or cryotherapy treatment and oral triazole compounds. Assistance is available through the CDC and, for Department of Defense beneficiaries, certain military facilities. PMID- 15757167 TI - A clinician's guide to managing Helicobacter pylori infection. AB - Helicobacter pylori infection is chronic and very common. Its clinical consequences vary widely, ranging from being asymptomatic and clinically insignificant in many cases to causing dyspepsia, peptic ulcer disease, and gastric malignancy in others. Care must be used in deciding whom to test for H pylori infection, as a positive result mandates treatment, making broad-based screening impractical. PMID- 15757168 TI - CT of the heart: principles, advances, clinical uses. AB - Computed tomography (CT) has become a standard test for many cardiovascular conditions (eg, aortic dissection and pulmonary embolism), and it has great potential in assessing other common diseases, including coronary artery disease. We review the principles of CT and its uses in cardiovascular medicine. PMID- 15757169 TI - New cervical cancer screening strategy: combined Pap and HPV testing. AB - Our strategy for cervical cancer screeing is being revolutionized by our new understanding of how human papillomavirus (HPV) contributes to carcinogenesis and the natural history of cervical cancer. The American Cancer Society and the American College of Obstetricians and Gynecologists now recommend combined HPV and Papanicolaou (Pap) testing for cervical cancer screening in women age 30 or older. However, although incorporation of HPV DNA testing into primary screening provides clear benefits, it also raises new questions. PMID- 15757170 TI - Overactive bladder: evaluation and management in primary care. AB - Primary care physicians must initiate a discussion of overactive bladder and urinary incontinence with their patients who are at risk. A stepwise approach to evaluation and diagnosis and the use of systematic evaluation and treatment algorithms suitable to the primary care setting will improve identification and effective management of the incontinent patient. PMID- 15757171 TI - Issues in anticoagulant therapy: recent trials start to answer the tough questions. AB - Recent trials provide evidence to help guide the management of patients with recurrent episodes of venous thromboembolism, cancer patients with venous thromboembolism, and patients receiving anticoagulant therapy who must undergo surgery. PMID- 15757172 TI - Fungal genetic resource centres and the genomic challenge. AB - Fungal research and education has for many years been supported by public service genetic resource centres, whose roles have been to maintain, preserve and supply living cultures to the research community. In the genomic era, genetic resource centres are perhaps more important than ever before. The cultures held, many of which are described and validated by expert biosystematists, are valuable resources for the future. There is a need to supply genomic and proteomic research programmes with fully characterised organisms, as usage of organisms from unreliable sources can prove disastrous, not least in economical terms. However, mycologists often require more than just the organisms, for example, their associated information is vital for bioinformatic applications and some researchers may only require genomic DNA from the organism rather than the organism per se. Genetic resource centres are continually adapting to meet the needs of their users and the wider mycological research community, this associated with OECD international initiatives should ensure they exist to support research for many years to come. This review considers the impact of such initiatives, the current roles of fungal genetic resource centres, the mechanisms used to preserve organisms in a stable manner and the range of resources that are offered for genomic research. PMID- 15757173 TI - Molecular phylogeny and taxonomy of the genus Pythium. AB - The phylogeny of 116 species and varieties of Pythium was studied using parsimony and phenetic analysis of the ITS region of the nuclear ribosomal DNA. The D1, D2 and D3 regions of the adjacent large subunit nuclear ribosomal DNA of half the Pythium strains were also sequenced and gave a phylogeny congruent with the ITS data. All the 40 presently available ex-type strains were included in this study, as well as 20 sequences of recently described species from GenBank. Species for which no ex-type strains were available were represented by either authentic strains (6), strains used in the 1981 monograph of the genus by van der Plaats Niterink (33), or strains selected on morphological criteria (17). Parsimony analysis generated two major clades representing the Pythium species with filamentous or globose sporangia. A small clade of species with contiguous sporangia was found in between the two main clades. A total number of 11 smaller clades was recognized, which often correlated with host-type or substrate and in several cases with a subset of morphological characters. Many characters used in species descriptions, such as antheridium position, did not correlate with phylogeny. A comparison of the ex-type and representative strains with all ITS sequences of Pythium in GenBank revealed limited infraspecific variation with the exception of P. rostratum, P. irregulare, P. heterothallicum, and P. ultimum. The total number of species examined was 116 (including 60 ex-type strains). Twenty six species had ITS sequences identical or nearly identical to formerly described species, suggesting possible conspecificity. The importance of comparing ITS sequences of putative new species to the now available ITS database in order to avoid unwarranted new species names being introduced. PMID- 15757174 TI - Molecular systematics of the Coronophorales and new species of Bertia, Lasiobertia and Nitschkia. AB - The Nitschkiaceae has been placed in the Coronophorales or the Sordariales in recent years. Most recently it was accepted in the Coronophorales and placed in the Hypocreomycetidae based on sequence data from large subunit nrDNA. To confirm and corroborate the taxonomic placement and monophyly of the Coronophorales, additional taxa representing the diversity of the group were targeted for phylogenetic analysis using partial sequences of the large subunit nrDNA (LSU). Based on molecular data, the Coronophorales is found to be monophyletic and its placement in the Hypocreomycetidae is maintained. The order is a coherent group with morphologies that include superficial, often turbinate, often collabent ascomata that may or may not contain a quellkorper and asci that are often stipitate and at times polysporous. Three species with accepted Nitschkia names, together with Fracchiaea broomeiana and Acanthonitschkea argentinensis, comprise the paraphyletic nitschkiaceous complex. Two new families, Chaetosphaerellaceae and Scortechiniaceae fams nov., are described for the clades containing Chaetosphaerella and Crassochaeta and the taxa having a quellkorper (Euacanthe, Neofracchiaea and Scortechinia) respectively. The Bertiaceae is accepted for the clade containing Bertia species. Three new species are described: Bertia tropicalis, Lasiobertia portoricensis, and Nitschkia meniscoidea spp. nov. PMID- 15757175 TI - Geographic isolation of Diplodia scrobiculata and its association with native Pinus radiata. AB - Diplodia pinea (syn. Sphaeropsis sapinea) is a well-known latent pathogen of Pinus spp. with a worldwide distribution. As such, this fungus is native where pines are endemic in the northern hemisphere and it has been introduced into all countries of the Southern Hemisphere where pines are exotic. The newly described D. scrobiculata (formerly known as the B morphotype of D. pinea) is thought to have a much more limited distribution. D. scrobiculata was first reported as an endophyte and weak pathogen of P. banksiana, where it was found to coexist with D. pinea. Diplodia scrobiculata is now known to have a much broader distribution in Northern America and Europe. In this study, seven Simple Sequence Repeat (SSR) markers were used to evaluate genetic diversity and gene flow between populations of D. scrobiculata. Results indicate a strong geographic isolation between populations of D. scrobiculata from different regions in North America, with unique alleles fixed in the different populations. The data fits the isolation by distance model indicating limited dispersal. Geographic isolation in combination with isolation by distance suggests prolonged reproductive isolation. Intensive collections of endophytes from native P. radiata in California have yielded only D. scrobiculata and not the significantly more pathogenic D. pinea. SSR analysis of three populations of D. scrobiculata from native P. radiata identified many shared alleles among the populations and moderate to high gene flow between them. The three Californian populations are distant and distinct from populations of D. scrobiculata from elsewhere. Under stress conditions, P. radiata is known to be very susceptible to D. pinea in plantations in the Southern Hemisphere. Native P. radiata is currently experiencing severe stress due to pitch canker caused by Fusarium circinatum. Such stress would provide ideal conditions for an associated outbreak of D. pinea. Thus, it is critical to prevent the movement of D. pinea into the last remaining native stands of P. radiata. PMID- 15757177 TI - The kinetics of 75[Se]-selenite uptake by Saccharomyces cerevisiae and the vacuolization response to high concentrations. AB - Uptake of 75[Se]-selenite by Saccharomyces cerevisiae has been characterized. At a 0.5 mM selenite, approximately 0.14 nmol Se (10(6) cells)-1 was rapidly accumulated by the cells at a rate of approximately 56 pmol x min-1 (10(6) cells) 1 which was independent of temperature and glucose. This rapid phase was followed by a slower uptake phase which was sensitive to glucose, temperature and metabolic inhibitors [2,4-dinitrophenol (DNP), carbonyl cyanide m-chlorophenyl hydrazone (CCCP), potassium cyanide (KCN) and sodium azide (NaN3)] and therefore presumed to be metabolism-dependent. Two transport systems appeared to be involved in selenite uptake. At the low range of selenite concentrations used (0.025-0.1 mM), a high affinity transport system occurred with apparent Km and Vmax values of 54.0 microM and 3.14 pmol x min(-1) (10(6) cells)(-1) respectively. A low affinity system was present at higher concentrations (0.1-1.0 mM) with apparent kinetic parameters of Km = 435 microM and Vmax = 11.6 pmol x min(-1) (10(6) cells)(-1). Elevated sulphate concentrations (up to 2.5 mM) did not affect the accumulation of selenite. However, the transport rate from 0.5 mM selenite was stimulated by sulphite, with the maximum effect occurring at 0.5 mM sulphite. Methionine had a detectable inhibitory action on selenite uptake whereas cystine and cysteine completely inhibited active transport of selenite. Transmission electron microscopy of 5 mM selenite-grown cells revealed the presence of abundant small cytoplasmic vesicles containing electron-dense granules which could represent an intracellular selenium-detoxification mechanism. PMID- 15757176 TI - Cloning and characterization of a gene rpg1 encoding polygalacturonase of Rhizopus oryzae. AB - The polygalacturonase (PG)-encoding gene (rpg1) of Rhizopus oryzae, the causal pathogen of rhizopus rot of mulberry, was cloned and sequenced. PGs were partially purified from incubation mixture of 2% pectin medium and their N terminal amino acid sequences were determined by a gas-phase protein sequencer. RT-PCR was performed using degenerate primers designed from the amino acid sequences, which resulted in part of a PG-encoding gene being obtained. By 3' RACE and TAIL-PCR analyses, the entire region of the PG-encoding gene was cloned and sequenced. The structural gene comprised 1199 bp coding for 383 amino acids with a putative signal peptide of 26 amino acids, and the open reading frame was interrupted by single intron of 47 bp. Phylogenetic analysis using the deduced amino acid sequence revealed that R. oryzae RPG1 belonged to a clade consisting of exo-PGs of ascomycete fungi. PMID- 15757178 TI - The assessment of the viability of the endospores of Rhinosporidium seeberi with MTT (3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide). AB - This report describes tests with Evan's Blue and MTT (3-[4, 5-dimethyl-2 thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide) for the assessment of the viability of rhinosporidial endospores. MTT stained a proportion of the spherical bodies that we regard as the Electron Dense Bodies (EDBs), and the cytoplasm of freshly prepared endospores or ones that were stored at 4 degrees. Slow-freezing at -20 degrees C, exposure to 10% formalin, or 0.1% sodium azide of the endospores abolished MTT-staining in both sites. Evan's Blue stained the EDBs and cytoplasm of fresh endospores or those stored at 4 degrees, and of sodium azide treated or frozen (-20 degrees)-thawed endospores. TMRE (tetramethyl-rhodamine ethyl ester) specifically labelled the spherical bodies, supporting the conclusion that these spherical bodies have a mitochondrial-like structure. TMRE staining was however retained in endospores after their treatment with formalin, sodium azide and slow-freezing while MTT-staining was abolished in all these treated endospores. These results indicate that EvB and TMRE were capable of revealing the morphological integrity of endospores but failed to identify the metabolic inactivation of the endospores after treatment with formalin, sodium azide or slow-freezing. Only MTT was capable of identifying metabolically active endospores and hence their viability and could prove to be of value in standardizing models of infection. PMID- 15757179 TI - The isoepoxydon dehydrogenase gene of patulin biosynthesis in cultures and secondary metabolites as candidate PCR inhibitors. AB - The European Union (EU) has introduced statutory limits for patulin in fruit products. Species definitions need to be unambiguous for this capability, especially to determine 'weak spots' in food, drink and feed production. Fungi were analysed for the isoepoxydon dehydrogenase (IDH) gene of the patulin metabolic pathway to indicate potential patulin production. In several cases inhibition of PCR product formation was indicated. Dilution as a means of overcoming PCR inhibition and to determine optimal concentrations was assessed and the ramifications for nucleic acid analysis in general are highlighted. The inhibitors may be secondary metabolites. However, inhibition was not involved obviously for Penicillium brevicompactum. The gene was detected frequently from Aspergillus section Clavati and Penicillium subgenus Penicillium species. Some strains within certain species were negative for the gene. Detection of the gene product in Byssochlamys is reported. PMID- 15757180 TI - Fire-related changes in biomass of hypogeous sporocarps at foraging points used by a tropical mycophagous marsupial. AB - Changes in pre- and post-fire biomass of hypogeous fungal sporocarps remaining at locations where an endangered mycophagous marsupial, the northern bettong (Bettongia tropica), had foraged, were assessed in fire-prone sclerophyll forest in northeastern Australia. Low to medium intensity experimental fires were set during the late dry season in 1995 and 1996 and post-foraging biomass of sporocarps (expressed as biomass of sporocarps remaining at recent B. tropica diggings) was measured at unburnt and burnt sites at approximately six-week intervals for a period of 14 months. Post-foraging biomass was significantly higher at burnt sites immediately following fire compared with control sites, solely due to increased biomass of hypogeous species belonging to the family Mesophelliaceae. Several months after fire, post-foraging biomass was significantly higher on unburnt sites compared with very low biomass on burnt sites. Twelve months after fire, the biomass on burnt and unburnt sites was not significantly different, having returned to biomass observed pre-fire. All evidence points toward mesophellioid fungi being greatly more available to bettongs on recently burnt ground, but fire may make several other sporocarp taxa considerably less available several months following fire. PMID- 15757181 TI - Interspecific interactions between the rare tooth fungi Creolophus cirrhatus, Hericium erinaceus and H. coralloides and other wood decay species in agar and wood. AB - Creolophus cirrhatus, Hericium erinaceus and H. coralloides were paired against over 20 other wood decay fungi from beech (Fagus sylvatica) covering a range of ecological strategies, on 2% malt agar (MA), 0.5% MA, 0.5% MA adjusted to -1.25 MPa by addition of KCl, 0.5% MA adjusted to pH 4 with KOH/H3PO4 and 0.5% MA under 5% O2/30% CO2 all at 20 degrees C. Creolophus cirrhatus and H. coralloides were also paired against 17 other species in wood, incubated at 20 degrees for 36-38 wk. They were average to good combatants, deadlocking with or replacing over half of the antagonists, H. erinaceus being slightly more combative than the other two species. Outcomes in wood were representative of those in agar. Abiotic conditions altered the outcomes slightly, reduced water potential favouring C. cirrhatus and H. erinaceus. Overall extension rate of C. cirrhatus and H. erinaceus often increased in the presence of antagonists. There was prolific production of fruit bodies in controls and interactions, though notably not under altered gaseous regime. Results are discussed in an ecological context. PMID- 15757182 TI - Termitomyces bulborhizus sp. nov. from China, with a key to allied species. AB - The new species, Termitomyces bulborhizus, is described from south-west China. The stipe of the basidiome is swollen into a prominent bulbous base at ground level, and this, together with the floccules on the stipe surface, distinguishes this species within the genus. T. bulborhizus is compared with closely related species, including three originally described from China, and a key to T. bulborhizus and related species is provided. PMID- 15757183 TI - Red-capped species of Russula sect. Xerampelinae associated with dwarf scrub. AB - The microscopic structure of herbarium material of alpine and arctic species of Russula sect. Xerampelinae from Europe and Greenland was studied. Based on our observations two taxa with prevailingly red pilei are accepted from the area: R. subrubens and R. pascua. Other related species occurring in arctic areas are R. cicatricata with a coppery coloured pileus and R. clavipes with a prevailingly greenish olivaceous pileus. As a consequence of the study of type material, R. chamiteae var. chamiteae and R. chamiteae var. microsperma are synonymized with R. subrubens. No European collections could be referred to R. oreina, and accordingly we suggest that this species is absent from Europe. PMID- 15757184 TI - A specific primer PCR and RFLP assay for the rapid detection and differentiation in planta of some Mycosphaerella species associated with foliar diseases of Eucalyptus globulus. AB - It is difficult to accurately identify Mycosphaerella species associated with leaf diseases of Eucalyptus based on morphological characters, as there is considerable overlap between very similar species and subspecies, and isolation from the host is not easy. Thus, a PCR and RFLP assay based on the ITS region of nr DNA was developed for the rapid detection and differentiation of M. nubilosa, M. cryptica and two non-sporing unidentified Mycosphaerella species isolated from the foliage of trees in resistant and susceptible families of E. globulus in a seed orchard at Kinglake West, Victoria, Australia. The M. nubilosa primer pair MNF/MNR was highly specific. A PCR-RFLP system based on the primer pair MCF/MCR, coupled with two restriction enzymes (DdeI and Tru1 I), differentiated M. cryptica, M. nubilosa, M. tasmaniensis and M. aff. vespa. One of the unidentified field-isolated Mycosphaerella species was identified as M. grandis on the basis of ITS sequence data while the other species remains unidentified. A PCR-RFLP system based on the primer pair U1F/U1R, coupled with the restriction enzyme StyI, differentiated between the two unidentified species. Unexpectedly, unlike isolation and culture studies, these assays detected M. nubilosa, M. cryptica and M. grandis in all single lesions examined on both juvenile and adult leaves, and on both highly resistant and highly susceptible E. globulus trees at this site. PMID- 15757185 TI - Sclerosing epithelioid fibrosarcoma: a case report. AB - Sclerosing epithelioid fibrosarcoma (SEF) is a rare variant of fibrosarcoma. It is a mesenchymal neoplasm composed of round to oval cells dispersed in a nestlike or cordlike distribution on a highly collagenous tissue background and is now recognized as a distinct clinical entity. In this report we discuss the presentation, diagnosis, and treatment of SEF. We focus on the unique histologic and immunohistochemical properties of this lesion. It is essential for both the surgeon and the pathologist to be aware of SEF and include it in the differential diagnosis for atypical head and neck neoplasms. Sclerosing epithelioid fibrosarcoma is a rare neoplasm that is now listed among the low-grade neoplasms that may occur in the head and neck. However, its behavior tends to be typical of more aggressive lesions. An awareness of this characteristic of SEF is essential for clinicians and pathologists alike. PMID- 15757186 TI - Embolization of an extracranial internal carotid artery pseudoaneurysm. AB - Deep neck space infections are rarely complicated by pseudoaneurysms of the extracranial internal carotid artery. This condition has a high mortality rate with conservative management and significant morbidity with open surgical techniques. Recent advances in endovascular therapy have allowed embolization of the diseased vessel to be performed with a more acceptable safety profile. We report a case of an extracranial internal carotid artery pseudoaneurysm complicating a deep neck space infection in an adolescent that was treated with selective endovascular embolization. We present the findings of computed tomography, magnetic resonance angiography, and conventional angiography. PMID- 15757187 TI - Supracricoid hemilaryngopharyngectomy in patients with invasive squamous cell carcinoma of the pyriform sinus. Part II: Incidence and consequences of local recurrence. AB - Supracricoid hemilaryngopharyngectomy (SCHLP) was performed in 147 patients over a 19-year period for previously untreated invasive squamous cell carcinoma of the pyriform sinus. With a minimum of 3 years' follow-up, the current retrospective series was designed to document the incidence, risk factors, and consequences of local recurrence following SCHLP. Before operation, 97.4% of patients had an induction chemotherapy regimen. A complete clinical response and a complete histologic regression were noted in 21.7% and 16.8% of patients, respectively. A significant statistical relationship (p = .0001) was noted between complete clinical response and complete histologic regression. Postoperative radiotherapy was used in 49.8% of patients. The overall local recurrence rate was 8.2%. The 5 year actuarial (Kaplan-Meier life-table method) local control estimate was 90.4%. As a function of T stage, the 5-year actuarial local control estimates were 96.2%, 91.1%, 92.9%, and 62.6% in patients with tumors classified as T1, T2, T3, and T4a, respectively. On univariate analysis, the overall local recurrence rate varied significantly, from 5.3% to 55.6% if the apex of the pyriform sinus was invaded (p = .02), 6.9% to 18.7% if the posterior pharyngeal wall was invaded (p = .03), and 6.3% to 60% if the margins of resection were positive (p = .02). In a stepwise regression model, positive margins of resection (odds ratio, 8.4; 95% confidence interval, 2.2 to 32.2; p = .002) and invasion of the apex of the pyriform sinus (odds ratio, 6.1; 95% confidence interval, 1.1 to 33.3; p = .04) were the variables statistically associated with an increased risk of local recurrence. Local recurrence resulted in a statistically significant increased risk of nodal recurrence (p = .005) and death (p < .004). The overall laryngeal preservation rate was 91.2%. From an oncological perspective, these results suggest that SCHLP should become a major tool in the armamentarium of the head and neck surgeon and should be integrated into future trials aimed at organ preservation in patients with invasive squamous cell carcinoma of the pyriform sinus. PMID- 15757188 TI - Combined horizontal and posterior canal benign paroxysmal positional vertigo in three patients with head trauma. AB - We report 3 patients who complained of positional vertigo shortly after head trauma. Positional maneuvers performed in the plane of the posterior canal (PC; Dix-Hallpike maneuver) and the horizontal canal (HC; patients were rolled to either side in a supine position with the head raised 30 degrees) revealed a complex positional nystagmus that could only be interpreted as the result of combined PC and HC benign paroxysmal positional vertigo (BPPV). Two patients had a right PC BPPV and an ageotropic HC BPPV, and 1 patient had a bilateral PC BPPV and a left geotropic HC BPPV. All 3 patients were rapidly free of vertigo after the PC BPPV was cured by the Epley maneuver and the geotropic HC BPPV was cured by the Vannucchi method. The ageotropic HC BPPV resolved spontaneously. Neuroimaging (brain computed tomography and/or magnetic resonance imaging scans) findings were normal in all 3 patients. From a physiopathological viewpoint, it is easy to conceive that head trauma could throw otoconial debris into different canals of each labyrinth and be responsible for these combined forms of BPPV. Consequently, in trauma patients with vertigo, it is mandatory to perform the Dix Hallpike maneuver, as well as supine lateral head turns, in order to diagnose PC BPPV, HC BPPV, or the association of both. Early diagnosis and treatment of BPPV may help to reduce the postconcussion syndrome. PMID- 15757189 TI - State-dependent laryngomalacia in sleeping children. AB - Laryngomalacia is a common congenital laryngeal abnormality. Despite its being widely discussed in the literature, the pathophysiology is not clearly understood. Both anatomic and neuromuscular theories have been suggested to explain laryngomalacia. We report 4 cases of laryngomalacia in which the presenting signs occurred during sleep. Awake flexible nasopharyngolaryngoscopy failed to demonstrate supraglottic structure collapse. Only while the patients were breathing spontaneously under general anesthesia was laryngomalacia noted. A proposed algorithm for diagnosis and treatment is included. These 4 cases of state-dependent laryngomalacia support a neuromuscular cause for laryngomalacia. PMID- 15757190 TI - Subtotal carbon dioxide laser arytenoidectomy for the treatment of bilateral vocal fold immobility: long-term results. AB - Sixty-nine patients underwent subtotal carbon dioxide laser arytenoidectomy for treatment of bilateral vocal fold immobility between 1985 and 2000. The population included 69 patients whose mean age was 56 years (SD, 16 years; range, 11 to 82 years). The mean follow-up was 50 months (SD, 44 months; range, 1 to 181 months). The overall postoperative peak expiratory/peak inspiratory flow ratio (normal value, 1) significantly improved (closer to 1; p = .0036). Voice analyses were also undertaken for 27 patients, almost exclusively after operation, given the context of initial emergency. The maximum phonation time averaged 6.57 seconds (median, 6 seconds). The phonation quotient remained high, with a mean of 503 (median, 440), and the mean conversational voice intensity remained around 59 dB. The median frequency analysis type was 3. The advantage of subtotal arytenoidectomy lies in the fact that it maintains a certain degree of rigidity along the posterior limit of the arytenoid frame, preventing inward collapse of the mucosa and thus lowering the risk of aspiration. PMID- 15757191 TI - Roles of free radicals, nitric oxide, and scavenging enzymes in nasal polyp development. AB - The aim of this study was to investigate the role of nitric oxide (NO), free oxygen radicals, and scavenging enzymes in the development of nasal polyp (NP) disease. This study included 41 patients who underwent endoscopic surgery because of NPs. Control specimens were taken from the inferior turbinate of 32 patients who underwent septoplasty. The levels of malondialdehyde (MDA), NO, and superoxide dismutase (SOD) were measured in intraoperative specimens of polyp tissue and turbinate mucosa. The levels of tissue NO were 191.06 +/- 26.62 micromol/mg of protein in patients with NPs and 145.30 +/- 19.19 micromol/mg of protein (p < .001) in the control group. The levels of MDA in the study and control groups were 12.47 +/- 2.12 nmol/mg and 8.83 +/- 1.08 nmol/mg (p < .01), respectively. The levels of SOD in the study and control groups were 50.77 +/- 14.74 U/mg and 77.93 +/- 15.31 U/mg (p < .001), respectively. It was determined that the levels of MDA in plasma and erythrocytes were higher in the patients with NPs than in the control group (p < .05). The levels of NO in plasma and erythrocytes in both groups were similar. The levels of SOD in plasma and erythrocytes were lower in patients with NPs than in the control group (p > .05). Increases in the levels of tissue MDA and NO and decreases in scavenging enzymes in patients with NPs as compared to control groups may indicate the presence of free radical damage in patients with nasal NPs. New studies are needed to clarify the efficacy of using antioxidants in the treatment of NPs. PMID- 15757192 TI - Loss of FHIT expression in squamous cell carcinoma and premalignant lesions of the larynx. AB - The tumor suppressor gene FHIT (fragile histidine triad) at chromosomal position 3p14.2 is altered by deletions in human tumors. The frequency and specificity of its inactivation vary among carcinomas, but few articles have referred to premalignant lesions such as dysplasia. We studied the expression of FHIT in a series of squamous cell carcinomas and premalignant lesions of the larynx. We observed 36 laryngeal carcinoma biopsy specimens and 70 dysplasia biopsy specimens. We studied FHIT expression in carcinoma and dysplasia with the immunohistochemical ABC (avidin-biotinylated peroxidase complex) method. Loss of FHIT protein was observed in 42% of the squamous cell carcinomas and 23% of the premalignant lesions. There was no significant difference among the three grades of dysplasia in FHIT expression. These findings of loss of FHIT protein expression, not only in squamous cell carcinoma, but also in premalignant lesions, indicate that FHIT alterations play an important role in the early events of carcinogenesis. PMID- 15757193 TI - Traction of lateral cricoarytenoid muscle for unilateral vocal fold paralysis: comparison with Isshiki' s original technique of arytenoid adduction. AB - Between 1995 and 1997, we performed Isshiki's original method of arytenoid adduction alone or as an adjunct to type I thyroplasty for the treatment of unilateral vocal fold paralysis. From 1997 onward, we performed arytenoid adduction by traction of the lateral cricoarytenoid muscle (Iwamura's method), because it reduces discomfort to the patient and avoids rotation of the thyroid cartilage. Preliminary experiments and surgical procedures involving traction of the lateral cricoarytenoid muscle are described. Of 21 patients with a maximum phonation time of less than 9 seconds, 14 underwent type I thyroplasty as an adjunct to our method of arytenoid adduction and 7 underwent arytenoid adduction alone. Sixteen patients (76%) were able after surgery to extend their maximum phonation time beyond 10 seconds; this result compares favorably with the results of Isshiki's original adduction technique. We describe useful anatomic landmarks for approaching the lateral cricoarytenoid muscle in the hope that more voice surgeons will adopt this approach in the treatment of unilateral vocal fold paralysis. PMID- 15757194 TI - Immunohistochemical study on distribution of mast cell phenotypes in human laryngeal mucosa: evidence for laryngeal type I allergy. AB - It is controversial whether or not type I allergic reactions can occur in the human laryngeal mucosa. To clarify this issue, we studied the distribution of mast cell phenotypes in the human laryngeal mucosa using the immunohistochemical staining method with antihuman tryptase and chymase antibodies. A large number of mast cells are present in the superficial layer of subepithelial connective tissue (SECT) of the epiglottis, arytenoid, and subglottis. Although mast cells containing both tryptase and chymase are predominant in the deep layer of the SECT, the majority of mast cells containing tryptase alone are located in both the epithelial layer and the superficial layer of the SECT. We conclude that the human laryngeal mucosa has the potential to induce type I allergic reaction. PMID- 15757195 TI - The normal-weight snorer: polysomnographic study and correlation with upper airway morphological alterations. AB - Obesity is recognized as playing an important role in causing snoring and in turning simple snoring into obstructive sleep apnea syndrome (OSAS). From our series of patients with sleep disturbances, we studied a group of 43 normal weight snorers in whom we detected a significant number of OSAS episodes. An articulated diagnostic protocol was adopted, and Muller's maneuver was extensively applied. The resulting data were compared to data from a group of 43 obese patients from the same series. The major risk factor for developing OSAS in normal-weight snorers appears to be anatomic abnormalities, in particular, septal deviation and base of tongue hypertrophy. Soft palate hypertrophy alone is not enough, although in obese snorers it can produce a sleep disorder. The normal weight snorer needs to be thoroughly investigated because of the significant risk of developing OSAS and for the detection of multiple concomitant sites of obstruction. PMID- 15757196 TI - Complications of mastoiditis in children at the onset of a new millennium. AB - The aim of the present study was to review our recent experience in the diagnosis and treatment of acute mastoiditis and its complications in a single tertiary care, university-affiliated pediatric center. Ninety-eight children with 101 episodes of acute mastoiditis were included in the study. The mean interval from onset of illness to mastoiditis was 4.5 days. Ear cultures most often grew Streptococcus pneumoniae and Pseudomonas aeruginosa (23.7% each). Complications occurred in 15.8% of episodes. The only factor differentiating children with and without complications was white blood cell count. These findings indicate that acute mastoiditis not only is a complication of prolonged infection of the middle ear, but may also present as an acute infection of the mastoid bone that can progress within 48 hours. The complication rate remains high, and antibiotic treatment at the onset of symptoms does not prevent complications. A high white blood cell count on admission may serve as a predictive factor of complicated cases. PMID- 15757197 TI - Audiological features and mitochondrial DNA sequence in a large family carrying mitochondrial A1555G mutation without use of aminoglycoside. AB - To elucidate the pathophysiological and genetic mechanisms of hearing loss associated with the homoplasmic mitochondrial A1555G mutation in the absence of aminoglycoside exposure, we conducted audiological and genetic analyses on 67 maternally related members of a large Japanese family carrying this mutation. A consistent pattern was evident in the audiograms, with features of sensory presbycusis, cochlear origin at all levels of hearing loss, and a high degree of vulnerability of outer hair cells. That the degree of hearing loss was similar in affected subjects within the same sibling group but differed between sibling groups suggests the involvement of nuclear modifier genes. Total mitochondrial DNA sequences were completely identical among subjects with various levels of hearing loss, and lacked additional pathogenic mutations. For the diagnosis of sensorineural hearing loss, the mitochondrial A1555G mutation should be considered when these features are present even in the absence of aminoglycoside exposure. PMID- 15757198 TI - Fine structure histopathology of labyrinthitis ossificans in the gerbil model. AB - Labyrinthitis ossificans (LO) is the pathological deposition of new bone within the lumen of the cochlea and labyrinth. This process occurs most commonly as a result of infection or inflammation affecting the otic capsule. Trauma and vascular compromise can also lead to neo-ossification within the otic capsule. The mechanism that regulates this process remains unestablished. This study details the end-stage histopathology in high-resolution plastic thin sections. Twenty Mongolian gerbils were infected by intrathecal injection of Streptococcus pneumoniae type 3 followed by subcutaneous penicillin G procaine (8 days) and were painlessly sacrificed 3 months later. The cochleas were serially divided and sectioned for light and electron microscopy. Sixteen of 20 animals (27 of 40 cochleas) demonstrated LO. Cochlear damage was most extensive in the vestibule and basal turn and decreased toward the apex, which often appeared normal. The histopathologic findings consisted of 1) new bone, calcospherites, osteoid, and fibrosis without dense connective tissue or osteoblasts extending from the endosteal wall into the lumen of the vestibule and scala tympani; 2) areas of dense connective tissue and osteoid enclosed by epithelial cells conjoined with the organ of Corti, stria vascularis, spiral ligament, and vestibular (Reissner's) membrane; and 3) partial to complete loss of the organ of Corti, spiral ligament cell bodies, stria vascularis, and spiral ganglion cells. Osteoblastic activity was not demonstrated in end-stage ossification in LO in the gerbil model. Neo-ossification appears to occur by calcospherite deposition along collagen-like fibrils within osteoid. The destruction of the organ of Corti, spiral ganglion cells, stria vascularis, and cells of Reissner's membrane and the spiral ligament occurs even in the absence of ossification of the cochlear duct. PMID- 15757199 TI - Experimental model of fungal sinusitis: a pilot study in rabbits. AB - We have established an experimental model of fungal sinusitis in rabbits to analyze the chronology and the pathogenesis of the development of noninvasive fungal sinusitis due to Aspergillus fumigatus. Thirty-four Pasteurella-free New Zealand white rabbits divided into three groups were included in this study. In the first group (10 rabbits), A fumigatus was inoculated into the maxillary sinus. In the second group (10 rabbits), A fumigatus was inoculated into the maxillary sinus in the presence of a wound in the mucosa. In the third group (14 rabbits), A fumigatus was inoculated into the maxillary sinus in the presence of a blocked ostium. On days 15 and 30, endoscopic, histopathologic, bacterial, and mycological examinations of both maxillary cavities and mucous membrane were performed. The rabbits were painlessly sacrificed 30 days after inoculation; mucosal and bone biopsies of the maxillary sinus cavities were performed for histopathologic studies. We found that noninvasive fungal sinusitis had been induced in 2 rabbits of the second group and 8 rabbits of the third group. We conclude that introduction of fungi into a sinus with a blocked ostium induces fungal sinusitis. The present model of experimental fungal sinusitis seems to be reproducible and suitable for further studies of the development of fungal sinusitis. PMID- 15757201 TI - Analysis of P161NK4A protein expression in follicular thyroid tumors. AB - PI6INK4A (P16) protein expression was analyzed immunohistochemically in archival material derived from 12 cases of follicular thyroid carcinoma, 57 cases of follicular adenoma and 17 cases of nodular goiter. Among follicular carcinomas, 11 out of 12 examined cases (91.7%) were positive for P161NK4A protein. Among follicular adenomas the percentage of immunopositivity was 76.5% (45/57) and among nodular goiter cases it was 19.3% (13/17). Overexpression of P16INK4A protein was found in 66.7% (8/12) of follicular carcinomas and in 19.3% (11/57) of follicular adenomas; the values of this parameter were statistically significantly higher in the follicular carcinoma group (p < 0.005). No P16INK4A protein overexpression was noted in nodular goiter cells. High immunohistochemically-detected expression of P16INK4A protein in follicular thyroid carcinoma cells suggests that the altered expression pattern of P16INK4A protein may disturb the regulatory mechanisms of thyreocyte cell cycle and plays a significant role in the formation of benign neoplasms and their malignant counterparts derived from follicular thyroid cell. PMID- 15757200 TI - Analysis of P53 and P21WAF1 proteins expression in follicular thyroid tumors. AB - The expression of P53 and P21WAF1 proteins was analyzed immunohistochemically in archival material derived from 12 cases of follicular thyroid carcinoma, 57 cases of follicular adenoma and 17 cases of nodular goiter. In the follicular carcinoma group 6 out of 12 cases (50%) were positive for P53 protein and 4 out of 12 cases (33.3%) were positive for P21WAF1 protein. In the follicular adenoma group, 18 out of 57 cases (31.6%) were positive for P53 and 16 out of 57 cases (28.1%) were positive for P21WAF1 protein. No positive cases of P53 or P21WAF1 proteins presence were found in the nodular goiter group. Positive correlation between the expression of P53 and P21WAF1 proteins was found for follicular carcinoma and adenoma groups (p = 0.034 and p = 0.002, respectively). The obtained results demonstrate that simultaneous immunohistochemical detection of P53 and P21WAF1 proteins expression may be useful in determining functional status of P53 protein, helping to interpret expression of P53 protein in thyroid follicular carcinoma cells. PMID- 15757202 TI - Analysis of nm23-H1 protein immunoreactivity in follicular thyroid tumors. AB - Immunohistochemical analysis employing a monoclonal antibody nm23-H1 (the antibody against nm-23 protein) was performed on archival material, consisting of 12 cases of follicular thyroid carcinoma (FTC), 57 cases of follicular thyroid adenoma (FTA) and 17 cases of nodular goiter (NG). Both cytoplasmic and nuclear immunoreactions for nm-23H1 were observed in cells of FTCs, FTAs and NGs. In oxyphilic adenomas cytoplasmic staining was observed. Eleven (91.7%) cases of FTC, 55 (98.2%) cases of FTA and 14 (82.4%) cases of NG were found to be positive for nm23-H1 protein. There were no statistically significant differences in the mean percentage values of immunopositive cells between carcinomas and adenomas. A significant increase in the number of cases with high percentage (more than 50) of positive cells was found in both carcinomas (FTCs) and adenomas (FTAs)--mainly microfollicular ones, in comparison with nodular goiter. It can be concluded that highly positive immunoreaction for the nm23-H1 protein in the cells of carcinomas (FTCs) and microfollicular adenomas indicates for a high proliferation rate of these tumors. PMID- 15757203 TI - Chlamydia pneumoniae infection in abdominal aortic aneurysm (AAA) patients and its clinical impact. AB - The aim of our study was to assess the presence of Chlmydia pneumoniae infection in AAA patients and to evaluate its association with clinical symptoms and histological signs of inflammation in the aortal wall. Fifty-two AAA patients participated in the research. Thirty healthy controls took part in serological examination. C. pneumoniae was detected by PCR and immunofluorescence in situ reaction in aorta samples of 84.6% and 86.54% of the patients, respectively. Serological markers of chronic C. pneumoniae infection were detected in 86.5% of AAA patients and in 33.3% of healthy controls. High titers of specific IgG and IgA were found in 37.8% of AAA patients with serologically defined chronic infection. All patients in "high serology" group had symptomatic aneurysm and inflammatory infiltrations in their aortal wall samples. AAA patients infected with C. pneumoniae are not a homogenous group. "High serology" group is much more prone to have symptomatic aneurysm than the remaining of AAA patients. Serology can be very useful in predicting the risk of AAA rupture. Inter-laboratory standardization of direct and indirect detection methods of C. pneumoniae infection is required to elucidate the role of these bacteria in AAA development. PMID- 15757204 TI - HER2 status in breast cancer determined by IHC and FISH: comparison of the results. AB - HER2 (human epidermal growth factor receptor 2) status became an important prognostic and predictive factor in breast carcinoma clinical management. There are two main techniques of evaluation of HER2 status: immunohistochemistry (IHC) for the protein expression and fluorescence in situ hybridization (FISH) for amplification of HER2 gene. The aim of the study was to compare the results obtained by IHC and FISH methods in determination of HER2 status in breast cancer. Three hundred and sixty breast cancer specimens were examined. Patients were operated in the Oncology Centre in Warsaw. IHC and FISH were performed in every case. IHC was performed with DAKO HercepTest and FISH with Oncor-QBiogene reagents. IHC results were classed into 4 groups, accordingly to the four-tier DAKO criteria system (0, 1+, 2+, 3+). FISH results were divided into three main categories: NA--no amplification, LA--low amplification and HA--high amplification. The number of copies of chromosome 17 was also assessed. Over 90% of cases described by IHC as 3+ exhibited amplification of HER2/neu gene. Remaining cases were positive with IHC, but presented no gene amplification. This might be due to the subjective assessment of the membrane staining. Another possibility is that overexpression of the protein was caused by mRNA stability or disorders in receptor degradation. The majority of cases classed by IHC as 2+ were also negative by FISH (80%). One fifth of IHC 2+ tumours were found to exhibit gene amplification. Remaining cases showed no amplification of HER2/neu gene, combined with aneuploidy of chromosome 17. All cases described by IHC as 0/1+ were also HER2-negative by FISH. IHC is well-established method of assessing HER2 status in breast cancer. Nonetheless, a group of cases described as 2+ should be additionally examined using FISH. The results obtained by the latter method are more reliable. In order to improve accuracy and gain the highest quality of HER2 status evaluation, in 2+ cases both methods should be applied. PMID- 15757205 TI - Angiodysplasia as a cause of recurrent bleeding from the small bowel in patients with von Willebrand disease. Report of 4 patients. AB - Angiodysplasia, characterized by the presence of malformed vessels in the submucosa of the gastrointestinal tract, may be a cause of recurrent bleeding. Bleeding angiodysplasia can be associated with von Willebrand disease (vWD) and this coincidence is probably the consequence of the lack of high molecular weight molecules of von Willebrand factor in the plasma. We report four patients with unexplained repeated massive intestinal bleeding, recurrent melena and iron deficiency anemia, which required numerous blood transfusions. All patients were adults (average age 68 years). Three patients have congenital von Willebrand disease (type 1, 2A and 3) and one idiopathic acquired von Willenbrand syndrome. Correct diagnosis was made 2-5 years after the onset of the symptoms and was confirmed by histopathological examination of surgically resected small bowel, where vascular lesions were located. Elderly patients with recurrent gastrointestinal bleeding and unexplained iron deficiency anemia should be diagnosed for angiodysplasia and vWD. PMID- 15757207 TI - Fundic gland polyps: sporadic or not sporadic, that is the question. PMID- 15757206 TI - Pneumatosis cystoides intestinalis--a report of two cases. AB - The authors present two cases of an extremely rare pneumatosis cystoides intestinalis of large and small intestine in a 48-year-old male and in a 77-year old female surgical patients. PMID- 15757209 TI - Current hazards in the healthcare workplace. PMID- 15757208 TI - Is it time for Kansas to pass new quarantine laws? PMID- 15757210 TI - Controlled substance prescriptions: the ARNP perspective. PMID- 15757211 TI - [Injury pattern on the hand after slipping onto the blade during a knife attack- a contribution to the differential diagnostic classification of sharp force injuries of the upper extremities]. AB - Differential diagnosis of injuries caused by accidental slipping of the hand onto the blade of a knife includes self-inflicted injuries (e.g. to simulate an offense) as well as active and passive defense injuries indicative of the involvement of another party. The injury pattern on the hands of five perpetrators who suffered accidental cuts during knife attacks, as seen by the authors during the clinical forensic examination, is presented. The injuries were localized on the ulnar aspect of the thumb facing the index finger, the radial aspect of the index finger facing the thumb and variable sites on the flexor or extensor side of the middle and ring fingers. Especially in those cases in which the injuries of the victim (e.g. involvement of bony structures) and the dominant hand of the perpetrator are known, additional information for the reconstruction of the course of events may be derived from the cut wounds occurring during the knife attack as the hand slipped onto the blade. PMID- 15757212 TI - [Assignment of a bite-mark on the victim's skin to the dentition of the perpetrator by means of anomalies in the number and position of the frontal teeth]. AB - This report is about a bite mark on the breast of a female corpse. The woman had been killed by ligature strangulation at the age of 76 years. The investigation of the dental casts of a suspected man showed some anomalies in the dentition and orthodontic variations. A comparison of the bite mark with these anomalies demonstrated the correspondence of several details. Consequently, the probability that the suspect was responsible for the bite mark was regarded as being very high. During the further investigations the suspect admitted the offence including the bite injury to the breast. The authors emphasize the importance of an exact photographic documentation as an indispensable prerequisite for an odontological comparison. PMID- 15757213 TI - [Visualization of bridging veins by means of postmortem computed tomography]. AB - At autopsy, visualization of lesions of the bridging veins, a frequent source of subdural bleeding, is difficult due to their anatomical localization. On the other hand their demonstration is of great importance for the assignment to a chronologically defined trauma. For this reason a postmortem method using computed tomography was developed to visualize the intracranial venous system by means of X-ray contrast media. In subdural bleedings, in which the skull had not been opened up, ruptured vessels could be accurately localized with this method, so that targeted dissection was possible during the subsequent autopsy. PMID- 15757214 TI - [Mandibular fracture following stab injury with a knife]. AB - Stab injuries of the face accompanied by fractures of the jaws are rare. The report deals with the case of a young man, who suffered a penetrating stab injury to the cheek and fracture of the mandible in an assault. The forensic questions to be answered were if the findings were compatible with a stab and subsequent fracture, if one of the confiscated instruments could have been the causative weapon and if the injury had to be assessed as life-threatening. Both the soft and the hard tissue injury confirmed the assumption that they were caused by a stab. It was not possible, however, to assign the injury to a specific knife from the submitted exhibits on the basis of the clinical findings. In the discussion the serious nature of stab injuries in the facial region is emphasized. PMID- 15757215 TI - [Initially suspected killing with a political background identified as accidental carbon monoxide poisoning]. AB - During a parish fair the police was called to a conflict between two rival right wing youth groups in the early morning hours. Shortly afterwards a burning car was discovered near the marquee with a charred male body on the driver's seat. It was assumed that there was a connection between the two events. At autopsy no injuries were found that were not attributable to postmortem consumption by the fire. The BAC value was 1.76 per thousand, the CO-Hb concentration 78%. The body could be identified by comparing pre- and postmortem X-rays of the molars and premolars of the right upper jaw. Reconstruction of the course of events and classification as an accident was possible only after calling in technical experts: The victim had gone back to his car to take a nap and kept the engine running because of the cold temperature. The car had been fitted improperly with a sport exhaust pipe, which caused both the carbon monoxide poisoning and the fire. PMID- 15757216 TI - [Investigations on the cause of death in a polytoxicomanic patient treated with wires and intermaxillary fixation for a chronic, infected mandibular fracture]. AB - The case of a 36-year-old male and former patient is presented who died for initially unknown reasons, with intermaxillary wire fixation for the treatment of an infected mandibular fracture still in place. The combined investigation of forensic experts and maxillofacial surgeons excluded aspiration and asphyxia as cause of death. Evidence was found for lethal methadone concentrations after intravenous drug abuse and a broncho pneumonia. Despite the clear evidence of a fatal intoxication in this case, caution must be exercised when dealing with individuals carrying intermaxillary fixation. PMID- 15757217 TI - [Progress in esophageal, stomach and large intestine cancers. Early forms can be endoscopically healed!]. PMID- 15757218 TI - [High-tech endoscopy and its limits. Utilizing progress but protecting patients from endoscopic undertreatment (interview by Dr. Peter Stiefelhagen)]. PMID- 15757219 TI - [Recurrent fecal blood. Sigmoidoscopy with erroneous conclusions]. PMID- 15757220 TI - [Patient is "intolerant" of certain foods. Validating an allergy diagnosis]. PMID- 15757221 TI - [Mercury as the suspected agent. Alzheimer disease due amalgam dental fillings? (interview by Dr. Judith Neumaier)]. PMID- 15757222 TI - [Varieties, pathological forms and unplumbed depths of an emotion. When loves makes you sick]. PMID- 15757223 TI - [Delusional jealousy and obsessive love--causes and forms]. AB - Obsessive love and delusional jealousy as persistent mental disorders, are well known but rare psychiatric conditions, the prevalence of which is estimated to be less than 0.1%. Delusional jealousy should not be confused with exaggerated "pathological" jealousy, which is characterized by excessive suspicion and possessiveness on the part of individuals who, as a rule, suffer from considerably disordered self-esteem. As a paranoid development, delusional jealousy occurs in particular among male chronic alcoholics. As in all delusional disorders, trivial events are evaluated with unshakeable certainty as proof of the correctness of the delusive notion. Obsessive love is seen predominantly in women. In the differential diagnosis, other mental disorders such as schizophrenia, manic or organic brain syndrome must be excluded. PMID- 15757224 TI - [Stalking: unwelcome company. The psychopathology of a modern phenomenon]. AB - The term stalking describes a pattern of behavior in which the victim is pursued, pestered and threatened. In many cases, the stalker resorts to physical violence, and may even commit murder. In the German-speaking areas, the phenomenon is to date not much discussed in the psychiatric and psychological literature, despite the fact that it is a widespread occurrence. While stalkers are diagnosed as suffering from schizophrenia and/or narcissistic as well as borderline disorders, victims have no typical "victim personality". They do, however, respond to stalking with anxiety, depression and post-traumatic stress disorders, which need to be recognized and treated. PMID- 15757225 TI - [Abnormal responses to the loss of a loved one]. AB - The loss of one's partner is always a stressful event. If this loss cannot be adequately coped with, depression, psychosomatic disorders or addictive disorders may result. In the worst case scenario, the victim of such a loss may resort to suicide or even kill the partner insisting on a separation. An active life, new personal contacts, or the support of a self-help group may help suffers to cope with the crisis more readily. Severe disorders--in part determined by the personality structure of the grieving partner--may require treatment by pharmaceuticals or cognitive therapy. PMID- 15757226 TI - [Soft-tissue tumors: primary excision or wait and see?]. AB - Soft tissue tumors are relatively commonly seen lesions in the doctor's office. An initial differentiation between malignant and benign tumors is usually possible on the basis of the case history and a careful physical examination. In adults, primary excision under local anesthesia is the treatment of choice in many cases, while in children and in the case of large tumors suspected of being malignant, a histological diagnosis must always be obtained. A number of procedures are available for biopsy taking. If the definitive histological work up confirms malignancy, or if the findings are uncertain, the diagnostic investigation should be extended to include such imaging procedures as CT scanning or MRI. PMID- 15757227 TI - [A leading symptom ignored. Many stroke patients experience pain]. PMID- 15757228 TI - [Even a physician is not safe from accident, stroke, etc.--caution: without power of attorney the general practice crumbles]. PMID- 15757229 TI - [Stress incontinence. How does the bladder become leakproof again?]. PMID- 15757230 TI - [Diagnostic quiz: Acute epigastric pain. The skin crackles the next day]. PMID- 15757231 TI - [10 minute consultation:36-year-old patient with chronic hepatitis C. Is unprotected sex with the spouse still possible?]. PMID- 15757232 TI - [Diagnosis at a glance. Completely without sports: upper arms like a bodybuilder]. PMID- 15757233 TI - Psychotherapy in a day clinic: results of a 1.5 year follow-up. AB - OBJECTIVE: In a prospective, naturalistic 1.5 year follow-up study of N= 114 consecutively admitted day clinic patients efficacy of the program and predictors of outcome are evaluated. METHODS: Patients had severe neurotic disturbances and personality disorders. Interviews and questionnaires (SCL-90-R, EDI) were used for evaluation at admission, discharge and follow-up. RESULTS: From 79.8% of the patients information could be obtained. In the main diagnostic categories between 30 to 50% of the patients showed complete remissions at follow-up. Patients at least kept improvements at follow-up. The rating of the "transition phase" after discharge was predictive for outcome. The group of patients that rated this phase as difficult showed higher levels of psychopathology at admission. CONCLUSIONS: It is possible to treat severely disturbed patients in a psychotherapeutic day clinic with good and lasting effects. A more disturbed group of patients needs special help to cope with the transition into the outpatient situation. PMID- 15757234 TI - Different traumatic experiences are associated with different pathologies. AB - We tested the hypothesis that different traumatic experiences will contribute in variable degree to different mental pathologies. A total of 223 young adult non patients were assessed with the help of self-reports. The role of six different trauma experiences (broken home, dysfunctional family, family violence, child sexual abuse, child severe sexual abuse and adult sexual abuse) in six different conditions/pathologies (alexithymia, depression, somatization, borderline, overall physical health and overall mental health) was tested in a series of multivariate analyses of variance and of Roy-Bargmann stepdown analyses. The hypothesis was confirmed: Individual traumatic experiences were indeed associated with different pathologies. Specifically, sexual abuse predicted borderline pathology, severe child sexual abuse somatization, and dysfunctional or broken family depression. Family violence was associated with worse overall mental health and alexithymia, whereas no trauma variable could be identified to be associated with overall physical health. Most of these individual relationships were reported in the literature, based on results obtained in different clinical samples. Our results were won in a sample of young non-patients controlling for overlap between pathologies. PMID- 15757235 TI - Ratings of early major depressive disorder symptom change during a brief psychiatric hospitalization. AB - Ratings of change in MDD severity during a brief psychiatric hospitalization were examined across informant sources to determine the extent of change from admission to discharge and if specific symptoms are especially likely to change. Study participants were 137 inpatients with a primary diagnosis of MDD. Symptom data were collected at admission and discharge from attending psychiatrists, nurses, and patients. Global ratings of MDD severity and specific MDD symptoms significantly decreased during the course of hospitalization. This effect held across informant sources. All symptoms were equally likely to change. Females were rated as more depressed at admission and discharge by psychiatrists, but no gender differences were seen in self-report or nurse ratings. Shorter length of stay and involuntary admission status were associated with greater reduction in MDD severity. The temporal course and magnitude of the symptom reduction may result in part from unique aspects of an inpatient setting or from an underreporting of symptoms. The association between a shorter length of stay and greater symptom reduction may reflect a distinction between treatment responders and nonresponders. PMID- 15757236 TI - Reducing restraints: alternatives to restraints on an inpatient psychiatric service--utilizing safe and effective methods to evaluate and treat the violent patient. AB - This paper describes the violence safety program instituted at Elmhurst Hospital Center in Queens, New York City in 2001, which significantly reduced the use of restraints and seclusion department wide, while providing a safe and therapeutic environment for patient recovery. The hospital service and program instituted is described, followed by restraint and seclusion data since 1998, and the program's results through 2003. Concurrent data in areas that could be affected by a reduction in restraint and seclusion such as self-injurious behaviors and altercations; use of emergency medication; use of special observation and length of stay data are also presented. In addition, types and frequency of alternative methods utilized to avoid restraints and seclusion are described. PMID- 15757237 TI - The safe treatment of the suicidal patient in an adult inpatient setting: a proactive preventive approach. AB - This paper focuses on the work of an inpatient service in an acute care safety net hospital in the prevention of suicide on its inpatient service and during the high-risk period post discharge. The strategy utilized a comprehensive proactive systems approach to the suicidal patient including: a formalized suicide assessment jointly done by medical and nursing staff, accurate diagnosis and best practice treatment, a flexible nursing observation policy, groups on inpatient units focused on suicide and key risk factors and triggers, and a communication tool to the next level of care that describes the strategies learned by the patient to deal with his or her suicidality. Incidents of self-injurious behaviors and special observation hours were monitored. PMID- 15757238 TI - Engaging patients and family members in patient safety--the experience of the New York City Health and Hospitals Corporation. AB - As one of its strategies to improve care, the New York City Health and Hospitals Corporation (HHC) has launched a number of initiatives to increase patient and family member involvement in the delivery of mental health services, including the hiring of peer counselors and the use of parent advocates. The effort to orient mental health services toward a more rehabilitative and recovery model has resulted in a significant change in the clinical culture. The article outlines patient and family member initiatives undertaken by the Corporation and offers some preliminary results that show the strong bond between increased patient and family member involvement and patient safety. PMID- 15757239 TI - Principal component for metabolic syndrome risk maps to chromosome 4p in Mexican Americans: the San Antonio Family Heart Study. AB - Metabolic syndrome refers to the clustering of disease conditions such as insulin resistance, hyperinsulinemia, dyslipidemia, hypertension, and obesity. To explore the genetic predispositions of this complex syndrome, we conducted a principal components analysis using data on 14 phenotypes related to the risk of developing metabolic syndrome. The subjects were 566 nondiabetic Mexican Americans, distributed in 41 extended families from the San Antonio Family Heart Study. The factor scores obtained from these 14 phenotypes were used in multipoint linkage analysis using SOLAR. Factors were identified that accounted for 73% of the total variance of the original variables: body size-adiposity, insulin-glucose, blood pressure, and lipid levels. Each factor exhibited evidence for either significant or suggestive linkage involving four factor-specific chromosomal regions relating to chromosomes 1, 3, 4, and 6. Significant evidence for linkage of the lipid factor was found on chromosome 4 near marker D4S403 (LOD = 3.52), where the cholecystokinin A receptor (CCKAR) and ADP-ribosyl cyclase 1 (CD38) genes are located. Suggestive evidence for linkage of the body size-adiposity factor to chromosome 1 near marker D1S1597 (LOD = 2.53) in the region containing the nuclear receptor subfamily 0, group B, member 2 gene (NROB2) also was observed. The insulin-glucose and blood pressure factors were linked suggestively to regions on chromosome 3 near marker D3S1595 (LOD = 2.20) and on chromosome 6 near marker D6S 1031 (LOD = 2.08), respectively. In summary, our findings suggest that the factor structures for the risk of metabolic syndrome are influenced by multiple distinct genes across the genome. PMID- 15757240 TI - Reproductive measures, fitness, and migrating Mennonites: an evolutionary analysis. AB - Given the same reproductive span, more children with shorter interbirth intervals and less parental attention per child should not do as well. There should be intermediate optima in family sizes, but only two studies have demonstrated optima. The goal here is to determine whether the relationship between fitness and fertility is linear and whether this relationship masks underlying variation in reproductive behaviors in a Mennonite congregation that lived in two disease settings, Prussia/Russia vs. Kansas. The relationships between children born and fitness were determined by calculating linear and quadratic regressions for total, Prussia/Russia vs. Kansas, and families with deaths vs. families with no deaths for total, Prussia/Russia, and Kansas. Variation was examined in terms of measures of reproductive success and reproductive span. Comparisons were made by t tests with Bonferroni correction. Regressions demonstrate equally well that the more children women bear, the higher the reproductive success, whether in the harsher disease ecology of rural Prussia/Russia or in less challenging rural Kansas and whether the women experience deaths or not. Prussian/Russian mothers bore significantly more children (6.5+/-0.3) than Kansan mothers (5.6+/-0.2) over longer reproductive spans but did not significantly increase the number of surviving children (4.9+/-0.2 vs. 4.7+/-0.2, respectively). Families experiencing deaths vs. no deaths exhibit significantly longer reproductive spans, reflecting a significantly earlier start at childbearing and a later finish, and produce significantly more children (5.4+/-0.2 vs. 4.2+/-0.2). Cox regressions were run, and the most significant covariates to negatively affect survivorship to 15 years were death in the family and length of the previous interbirth intervals. There was variation in families, but perhaps most had adequate nutrition, which may explain the lack of optima in fitness. PMID- 15757241 TI - Local extinction and recolonization, species effective population size, and modern human origins. AB - A primary objection from a population genetics perspective to a multiregional model of modern human origins is that the model posits a large census size, whereas genetic data suggest a small effective population size. The relationship between census size and effective size is complex, but arguments based on an island model of migration show that if the effective population size reflects the number of breeding individuals and the effects of population subdivision, then an effective population size of 10,000 is inconsistent with the census size of 500,000 to 1,000,000 that has been suggested by archeological evidence. However, these models have ignored the effects of population extinction and recolonization, which increase the expected variance among demes and reduce the inbreeding effective population size. Using models developed for population extinction and recolonization, we show that a large census size consistent with the multiregional model can be reconciled with an effective population size of 10,000, but genetic variation among demes must be high, reflecting low interdeme migration rates and a colonization process that involves a small number of colonists or kin-structured colonization. Ethnographic and archeological evidence is insufficient to determine whether such demographic conditions existed among Pleistocene human populations, and further work needs to be done. More realistic models that incorporate isolation by distance and heterogeneity in extinction rates and effective deme sizes also need to be developed. However, if true, a process of population extinction and recolonization has interesting implications for human demographic history. PMID- 15757242 TI - Ineffective contraceptive use and its causes in a natural fertility population in southern Jordan. AB - Based on our interview survey of 574 randomly selected married women from a rural population in the South Ghor district, Jordan, where traditional Arab customs have been persistently maintained, in this paper we analyze the age-specific marital fertility rates (ASMFRs) and contraceptive practices, especially the prevalence and duration of contraceptive use. The ASMFRs fitted the natural fertility pattern proposed by Coale and Trussell's model, and the total fertility rate was estimated to be 7.2. Even though the prevalence rate of modern contraceptive methods has reached 14.3%, because of the recent increase among young women in particular, the users had larger numbers of children than the nonusers and the duration of contraceptive use was short, especially at young ages (e.g., about 90% 24-month discontinuation probabilities in 15-19- and 20-24 year-olds). These ineffective contraceptive uses were related to traditional Arab norms, represented by the pooled proportion of "as many as possible" and "up to God" answers to the ideal number of children (70% of men and 30% of women). PMID- 15757243 TI - Genetic profile of the Kichwas (Quichuas) from Ecuador by analysis of STR loci. AB - Allele frequency data for the STR systems CSFaPO, TPOX, TH01, D13S317, D16S539, D5S818, and D7S820 were determined in a population sample of unrelated, healthy Amerindian Kichwa individuals. All loci met Hardy-Weinberg expectations, and there was no evidence for association of alleles among the seven loci. PMID- 15757244 TI - A common haplotype associated with the Basque 2362AG --> TCATCT mutation in the muscular calpain-3 gene. AB - Limb-girdle muscular dystrophy type 2A (LGMD2A) is caused by any of over 150 mutations in the calpain-3 (CAPN3) gene. Of those, 2362AG --> TCATCT is particularly prevalent in Basque patients, and this mutation was hypothesized to have arisen in the Basque Country. To explore the natural history of this mutation, we genotyped 65 Basque and non-Basque patients with LGMD2A who carry the 2362AG --> TCATCT mutation for four microsatellites within or flanking the gene. A particular haplotype was found in three-fourths of the patients and was assumed to be ancestral. From the average number of recombinations and mutations accumulated from this ancestral haplotype, the age of the 2362AG ----> TCATCT mutation was estimated to be 50 generations (i.e., 1,250 years), which is more recent than the Paleolithic Basque heritage. The subsequent spread of the 2362AG -> TCATCT mutation can be related to gene flow out of the Basque Country, even across a cultural border. PMID- 15757245 TI - Minimal sharing of Y-chromosome STR haplotypes among five endogamous population groups from western and southwestern India. AB - We attempt to address the issue of genetic variation and the pattern of male gene flow among and between five Indian population groups of two different geographic and linguistic affiliations using Y-chromosome markers. We studied 221 males at three Y-chromosome biallelic loci and 184 males for the five Y-chromosome STRs. We observed 111 Y-chromosome STR haplotypes. An analysis of molecular variance (AMOVA) based on Y-chromosome STRs showed that the variation observed between the population groups belonging to two major regions (western and southwestern India) was 0.17%, which was significantly lower than the level of genetic variance among the five populations (0.59%) considered as a single group. Combined haplotype analysis of the five STRs and the biallelic locus 92R7 revealed minimal sharing of haplotypes among these five ethnic groups, irrespective of the similar origin of the linguistic and geographic affiliations; this minimal sharing indicates restricted male gene flow. As a consequence, most of the haplotypes were population specific. Network analysis showed that the haplotypes, which were shared between the populations, seem to have originated from different mutational pathways at different loci. Biallelic markers showed that all five ethnic groups have a similar ancestral origin despite their geographic and linguistic diversity. PMID- 15757246 TI - Characteristic beta-globin gene cluster haplotypes of Evenkis and Oroqens in north China. AB - Haplotype frequencies of the beta-globin gene cluster were estimated for 114 Evenkis and 81 Oroqens from northeast China, and their characteristics were compared with those in Japanese, Koreans, and three Colombian Amerindian groups of South America (Wayuu, Kamsa, and Inga tribes). A major 5' subhaplotype (5' to the delta-globin gene) was + - - - - in Evenkis, whereas + - - - -, - + + - +, and - + - + + were the major subhaplotypes in Oroqens. One possible candidate for an ancestral 5' subhaplotype, - - - - -, was found in one Evenki (0.5%) and three Oroqen chromosomes (2.0%). They were observed as heterozygous forms for + ---- and -----. Major haplotypes were +-----+, + -----+-, and + - - - - + + in Evenkis, whereas they were +-----+,-++-+-+, +----+-, and -+-++-+ in Oroqens. The lowest Nei's genetic distance values of Evenkis or Oroqens based on the 5' subhaplotype frequency distributions were observed in relation to the Wayuu or Koreans, respectively, but those of Evenkis and Oroqens based on the haplotype frequency distributions were found in relation to Koreans. PMID- 15757247 TI - Hemoglobinopathies, G6PD deficiency, and hereditary elliptocytosis in Bahrain. AB - The native population of Bahrain has a high prevalence of hemoglobinopathies and G6PD deficiency, probably as a result of past malarial endemism. We used the Biorad-Variant hemoglobin testing system for primary screening of hemoglobinopathies in 20,000 individuals. Hemoglobin abnormalities were detected in 7,206 (36.3%) cases. PMID- 15757248 TI - D17S30 and TP53 VNTR loci polymorphisms in a Turkish population. AB - VNTR loci are useful as genetic markers in population genetics, forensics, and chimerism detection because of their high level of variability. In this study we present frequency information for two polymorphic VNTR loci, namely, D17S30 and TP53, which we examined in 100 healthy individuals from a Turkish population. PMID- 15757249 TI - A novel tetrameric short tandem repeat located in the 3' flanking region of the human ABO-secretor gene (FUT2) and association between FUT2 and FUT2/01 loci. AB - We found a novel polymorphic short tandem repeat (FUT2/01), 3.8 kb downstream of the coding region of FUT2. Seventeen length and 33 sequence variants were identified in 300 individuals representing three major human populations. Africans (Xhosa) and Europeans were characterized by high microvariation, and Japanese were characterized by a simple repeat structure. All exhibited high haplotype diversity. PMID- 15757251 TI - [Contemporary interventional pulmonology]. PMID- 15757250 TI - The need for the consideration of an Order of Pastoral Care: a proposal with responses. AB - The author notes that historically the pastoral care community was seen as a product of the pastoral care organizations. Those pastoral care organizations were actually dynamic products of the emerging pastoral community. Unfortunately, this community was not nurtured beyond the gathering of certified supervisors, counselors, and chaplains. The author contends that the establishment of the College of Pastoral Supervision and Psychotherapy (CPSP) was an attempt to correct this oversight. Thus, he suggests, it is compelling to consider the Pastoral Care Chapter as a microcosom of a possible Order of Pastoral Care. Responses to this suggestion by pastoral care colleagues accompanies the article. PMID- 15757252 TI - [Transbronchial lung biopsy as a diagnostic method of diffuse pulmonary diseases]. AB - The aim of the study was to assess the diagnostic yield of transbronchial lung biopsy (TBLB) in some of diffuse pulmonary diseases, like: sarcoidosis, organizing pneumonia--(OP), tuberculosis and neoplasmatic infiltrates of the lung. Transbronchial lung biopsy was performed in 123 patients, preceded by high resolution computed tomography (HRCT). The HRCT guidance helped to select the area of lung to be biopsied. In 80 patients (65%) TBLB enabled to diagnose 40 cases of sarcoidosis, 15 cases of OP, 13 cases of neoplasmatic infiltrates of the lung and 5 cases of tuberculosis. There were 43 cases undiagnosed by means of TBLB. In 19 patients the diagnosis was established by means of other methods like videothoracoscopy (8 cases), bronchoscopy with bronchial mucose biopsy (7 cases of sarcoidosis), mediastinoscopy (2 cases of Hodgkin's disease), transthoracic needle biopsy (2 cases of adenocarcinoma). Moreover sarcoidosis was diagnosed in 15 cases by means of clinical, radiological examination and BAL. 9 patients didn't agree for further invasive diagnostics. Transbronchial lung biopsy was shown to be efficient diagnostic method especially in sarcoidosis, OP and neoplasmatic infiltrates of the lung. However, in approximately 35% of cases of diffuse pulmonary diseases this technique doesn't allow to establish a diagnosis. This in turn implicates the necessity for further diagnostic procedures including videothoracoscopic or open lung biopsy. PMID- 15757253 TI - [Risk factors for asthma in children in Lodz region]. AB - The incidence of asthma and other allergic diseases continues to increase. In addition to genetic factors, environmental influences are thought to play an important role. The aim of this study was to identify factors that influence and drive the atopic march from atopic sensitization to asthma in children from Lodz. METHODS: 800 atopic children, aged 5-18 years, were included to our study. Parents filled in questionnaires and gave interviews about their children's diseases. 405 (43%) children have diagnosis of asthma. RESULTS: A significant association was observed between asthma and male sex, parents' history of atopy, parental highest school grade, maternal smoking during pregnancy, maternal chronic disease (especially chronic renal diseases), maternal allergen sensitizing diet during breast-feeding, increased exposure to indoor humidity and moulds. Similar effect was seen for episodes of wheeze occurring in the first 3 years of life as followed: wheezing during an airway infection, wheezing not connected with respiratory tract infection, wheezing not related with physical exercise. Child's daycare attendance (nursery school) was associated with decreased risk of asthma. PMID- 15757254 TI - [Fine-needle aspiration biopsy under CT-guidance--usefulness in diagnosis of malignant lung tumors]. AB - Aim of the study was to evaluate the usefulness of fine-needle aspiration biopsy (FNAB) in diagnosis of malignant lung tumors. MATERIAL AND METHODS: The results of 438 CT guided FNAB of lung lesions performed between Sep. 1st 1999 and Feb. 20th 2004 were evaluated. RESULTS: In 295 cases (67.35%) the cytological diagnosis of lung tumours were established by FNAB. The most common type of the tumours were: non-small cell lung cancer: 222 (50.69%) and small cell lung cancer: 49 (11.19%). 104 of these results were histologically verified and two false positive results were detected. In 143 cases (32.65%) however, the results established by FNAB were non-diagnostic. 63 of the lesions were verified and in 36 cases (57.1%) the malignant tumours were proved. The specificity of the method regarding malignant lesions reached 93.10%, and its sensitivity for patients with malignancy was 73.91%. The average diameter of the malignant tumour was 5.4+/-1.3 cm whereas in case of non-diagnostic results the diameter of the tumour was 4.1+/ 1.6 cm. However, there was no significant difference between the two groups. The most common complication of FNAB was pneumothorax, which occurred in 22 patients (5.02%) and in 13 cases (2.97%) required drainage of the chest cavity. CONCLUSION: FNAB is a safe diagnostic method with high specificity and moderate sensitivity in detecting of malignant lung lesions. PMID- 15757255 TI - [The effect of asthma and COPD exacerbation on exhaled nitric oxide (FE(NO))]. AB - Exhaled nitric oxide is a marker of airway inflammation and it is significantly decreased by glucocorticosteroid therapy, especially in patients with asthma. AIM OF THE STUDY: Evaluation of changes in FE(NO) in asthma and COPD exacerbation. MATERIALS AND METHODS: 17 patients with acute asthma and 19 patients with an exacerbation of COPD were enrolled to the study. FE(NO) (chemiluminescence, on line, restricted breath technique measurement in accordance with the ATS recommendations) was performed for five consecutive days following admission to hospital. Results of the following additional blood investigations: peripheral white blood cell count, ESR, C-reactive protein level, arterial blood gases, spirometry or peak expiratory flow were also analyzed. RESULTS: The average value of FE(NO) on admission was 41.5+/-10.7 ppb (95% CI: 18.8-64.2 ppb) asthma patients and 28.6+/-5.4 ppb (95% CI: 17.4-40.0 ppb) in COPD patients. In asthma patients a significant decrease of FE(NO) on the third day of therapy was observed (41.5 vs 26.1 ppb, p < 0.05). We found a positive correlation between FE(NO) on admission and the peripheral blood eosinophil count. In COPD patients a significant decrease of FE(NO) on the 4th day was noted (28.6 vs 17.5 ppb, p < 0.05). FE(NO) in both groups was higher than that of 19 healthy volunteers previously studied in our laboratory (14.1+/-4.7 ppb; 95% CI: 11.8+/-16.4 ppb). CONCLUSIONS: Exacerbations of asthma and COPD are associated with an increased FE(NO). FE(NO) measurement is a useful tool in the assessment of treatment efficacy. Exhaled nitric oxide may indicate the intensity of allergic inflammation in patients with asthma. PMID- 15757256 TI - [Granular cell tumor--a rare, benign respiratory tract neoplasm in the material of the Institute of Tuberculosis and Lung Diseases]. AB - The granular cell tumor (GCT) is a nodule that arises most commonly in the skin, the breast or the tongue. The vast majority are benign. Approximately 6-10% of granular cell tumors have been reported in the lower respiratory tract. The clinical, pathological and immunohistochemical findings of eleven cases are described in our material consisted of 6 males and 5 females aged from 35 to 58 years (median, 46 years). The GCT were solitary lesions in all our patients. The tumors were located in trachea (6 cases) and in bronchus (5 cases). They were found during bronchoscopy performed because of symptoms of pneumonia, lung cancer and hemoptysis or dyspnea alone. Diameter of the tumors ranged from 0.2-2.5 cm (median 1.2 cm). Six tumors were surgically excised and 5 were endoscopically removed. Pulmonary GCT behave in a benign fashion. It was observed that tumors of less than 8 mm were more amenable to endoscopic removal and larger tumors were more likely to infiltrate through the bronchial wall. Histologically, the GCT showed submucosal infiltrates of round or oval cells with abundant granular cytoplasm. The tumors cells were positive for S-100 protein, neuron specific enolase, CD68 and vimentin. Our immunohistochemical results are consistent with this concept. PMID- 15757257 TI - [Retrospective analysis of the results of Allergic Diseases Prevention Program in children in the Lodz area]. AB - Growing prevalence of allergic diseases requires concentrating on patients' education, prevention, suitable organization of medical care as well as the early diagnosis. It the presented retrospective study of the results of Prevention Allergic Diseases Programme in children in Lodz region in years 2000-2003 was analysed. Analysis of medical records of patients undergone preventive program showed: solid growth of number of patients with allergic disease in frames of Programme, the solid growth of percentage of new diagnosed allergic diseases, essential changes in structure of diagnosed allergic diseases during Programme, particularly visible in the youngest age group up to 5 years of age as well as the systematic fall in number of hospitalization from asthma and atopic dermatitis. With attention on growing number of referred patients Program should be continued and with the reason of still missing good medical care in children of Lodz region. PMID- 15757258 TI - [Effect of some demographic and social factors on the degree of nicotine addiction and motivation to quit smoking in healthy people]. AB - AIM: Estimation of the influence of some demographical and social factors on degree of nicotine addiction and motivation to quit smoking. MATERIAL AND METHOD: 208 healthy, smoking people entered the study. The Fagerstrom test was used for analyzing degree of nicotine addiction (heavy addiction > 7 points). Motivation for smoking cessation was estimated by Schneider test (high motivation > 5 positive answers). Tests results were analyzed in connection with age, gender, education and intensity of nicotine addiction defined as pack-years. RESULTS: In 37% of patients (77/208) heavy nicotine addiction was diagnosed. High motivation for quitting smoking was recognized in 74% people (150/208). Subjects with wild nicotine addiction had significantly higher motivation for smoking cessation (chi2, p=0.002). There was not influence of gender, age, education and addiction intensity (defined as pack-years) on the grade of nicotine addiction. We didn't notice relation between gender and motivation to quit. Young, well educated patients are significantly stronger motivated for smoking cessation. CONCLUSIONS: There was not direct dependence between the number of smoked cigarettes and degree of nicotine addiction. In the group of healthy subjects, young, well educated people have a better chance for smoking cessation. PMID- 15757259 TI - [Level of mannose-binding lectin (MBL) in patients with tuberculosis]. AB - Mannose-binding lectin (MBL) constitutes an important element in innate immune response against invading microorganisms. On the other hand, it has also been suggested that MBL, due their opsonic activity, might facilitate the uptake of certain intracellular pathogens. The aim of this study was to investigate whether MBL plays a role in tuberculosis. MBL serum concentration was determined in 68 patients with tuberculosis (TB), 30 patients with non-TB lung diseases and in 64 healthy controls. Serum MBL level was tested by an ELISA test with S. cerevisiae mannan-coated microtiter plates. Generally, TB patients had significantly higher MBL levels than both control groups, and in TB group percentage of individuals with highest MBL level was significantly higher (39.7%). These results indicate that high levels of MBL may be involved in the pathogenesis of tuberculosis and to be a relative disadvantage in mycobacterial infection. PMID- 15757260 TI - [IgG anti-multiantigen sonicate of Mycobacterium tuberculosis (Mts) measured by ELISA-Mts]. AB - Laboratory diagnosis of tuberculosis (TB) is often difficult. In recent years the detection of TB by serologic means has been the subject of a number of investigations. We studied the diagnostic utility of enzyme-linked immunosorbent assay in pulmonary tuberculosis. Two serological tests, commercial Pathozyme-Myco IgG (Omega Diagn.), and self-prepared ELISA-Mts using mycobacteria sonicate were evaluated simultaneously with 132 serum samples. Frequency and level of anti mycobacterial IgG were compared in sera from 68 patients with TB and in sera from 64 healthy volunteers. The sensitivities of the tests with sera from TB patients were, respectively 55.9% and 70.6%. The specificities of these tests were respectively 98.5% and 84.4%. By combining two tests, a maximum sensitivity of 76.5% was obtained with reciprocal drops in specificity to 82.8%. Despite that, it has been suggested that combinations of different antigens were attempted in order to improve the diagnostic yield. PMID- 15757261 TI - [Cystic fibrosis diagnosed in 36-year old man]. AB - Cystic fibrosis (CF) is the most common autosomal recessive disorder in Caucasians. There is considerable variability in the clinical presentation and course of cystic fibrosis. Adult patients with mild symptoms are increasingly being diagnosed with CF. We present a case of a man diagnosed with CF at age 36 years. The diagnosis was suspected because of recurrent nasal polyposis, bronchiectases, male infertility, and a positive sweat tests. One CFTR mutation was identified. CONCLUSIONS: 1. Cystic fibrosis should be included into the differential diagnosis of chronic respiratory symptoms in adults. 2. CF patients diagnosed in adulthood usually have milder clinical course of the disease. PMID- 15757262 TI - [Exogenous lipoid pneumonia--case report]. AB - A 72-year-old patient with a rare disease-exogenous lipoid pneumonia (ELP) was described. One year ago he underwent laryngectomy because of larynx carcinoma and now he was admitted to hospital because of suspicion of cancer progression in the lungs. Chest HRCT and microscopic picture of BALF suggested diagnosis of lipoid pneumonia. His history supported it because he used oil drops to hist tracheostomy outline. PMID- 15757263 TI - [Alveolar proteinosis after professional exposure to cotton and linen dust, successfully treated with whole lung lavage--a case report]. AB - Alveolar proteinosis can be primary or secondary. The secondary alveolar proteinosis was observed in infections, in malignancies and after exposition to various dusts. We present a 55 year old woman with severe pulmonary alveolar proteinosis, who was exposed to cotton and linen dust in work for 20 years. The main features were slowly increasing dyspnoea and cough. Chest X-ray revealed diffuse bilateral acinar infiltrates. We observed decreased diffusing capacity, and severe hypoxaemia. The diagnosis was confirmed by transbronchial lung biopsy. Histopathology examination revealed characteristic alveolar filling by PAS positive material. The patient underwent the whole lung lavage (WLL) with very good therapeutic effect. WLL was performed under general anaesthesia with a double-lumen endotracheal tube. We used 20 L 0.9% saline to lavage one lung. Clinical and functional improvement has persisted for over 1 year. PMID- 15757264 TI - [Diagnostic difficulties in primary mesothelioma]. AB - A 54-year-old woman with a history of fatigue and shortness of breath was found to have a pericardial effusion and mild mediastinal lymphadenopathy. Video assisted pericardioscopy revealed thickened pericardium studded with multiple nodules. Histologically the tumor was diagnosed as papillary adenocarcinoma. The site of the primary tumor could not be identified. As lung cancer is one of the most frequent causes of pericardial metastases the patient was treated with cisplatin and vinblastin. Following 5 courses of chemotherapy--given over a 4 month period--the amount of pericardial effusion and pericardial thickness did not change. The material from pericardial biopsy was reexamined and positive immunostaining for calretinine was found. The final diagnosis was primary pericardial mesothelioma of epithelioid type. Palliative radiotherapy of mediastinum was planned but the patient deteriorated and died due to disease progression with venous thrombosis and superior vena cava syndrome. The case illustrates the difficulties in establishing diagnosis of primary pericardial mesothelioma which is a rare tumor with poor prognosis. PMID- 15757265 TI - [Inhaled glucocorticosteroids in the treatment of asthma and chronic obstructive pulmonary disease]. AB - Molecular mechanisms of actions, pharmacokinetics and anti-inflammatory potency of inhaled glucocorticosteroids (ICS) are described. Differences in clinical effectiveness of ICS in asthma and chronic obstructive pulmonary disease (COPD) therapy and up-to-date recommendations for treatment with ICS in asthma and COPD patients are discussed. Finally there are presented data on safety of long-term treatment with ICS, specially in asthmatic children. PMID- 15757266 TI - [Non-specific humoral immunity in pathogenesis of tuberculosis]. PMID- 15757267 TI - [Hypersensitivity pneumonitis recognised by open lung biopsy in patients at the Institute of Tuberculosis and Lung Diseases]. AB - Diagnosis of hypersensitivity pneumonitis (HP) is made on the basis of a combination of clinical features connected with the exposition to organic dusts, radiographic abnormalities, lung function tests and immunological tests. Open lung biopsy (OLB) and histological examination is not necessary and is indicated mainly in chronic and subacute form of HP. In the chronic form of HP symptoms often do not show a temporal relationship with antigen exposure and errors occur especially if specific questions are not asked about exposure to antigen if HP. The aim of this paper was to analyse whether OLB was necessary in all patients in whom HP was recognised just after this procedure. Material included 21 patients in whom microscopic examination suggested HP among 258 pts in whom OLB was performed in the period of 1998-2003. There were 13 men and 8 women at the age of 18 to 65 years, mediana 42 years. RESULTS: We found 3 groups of pts. Group "+" included 9 exposed pts. They were asked and confirmed contact with birds, hay and 7 of them observed correlation between exposition and symptoms. The second Group "-" included 7 pts who were asked about exposition and who answered in the negative. In all of them HRCT scans suggested HP. Restriction was below 70% of VC in 3 pts. The third Group "0" included 5 pts who were not asked about exposition and correlation with symptoms. Two from them lived in the rural area. CONCLUSIONS: Open lung biopsy was not necessary for 6 patients in Group "+", because then had enough symptoms to recognise HP. In 3 patients of Group "+" and in 5 of Group "0" OLB could be not necessary because specific anamnesis and additional examinations were not collected. OLB was necessary method for 7 patients in Group "-". PMID- 15757268 TI - [Effect of montelukast on lung function and clinical symptoms in patients with cystic fibrosis]. AB - Inflammatory process contributes to progressive lung tissue damage in cystic fibrosis (CF). Cysteinyl leukotrienes have been found in the sputum of CF patients at concentrations sufficient to cause potent biological effect. This study was designed to assess the effect of anti-inflammatory treatment with montelukast sodium in CF patients. Twelve patients, aged 6-29 were recruited. It was 20 week, placebo-controlled, and randomized, double blind, crossover trial. At first and last week of each treatment course spirometry and whole body plethysmography parameters (FEV1, PEF, FEF25/75%, VC, TGV, Raw and RV) and clinical wheezing and cough scale were measured. In montelukast group significant improvement in FEV1 (mean +/- SD, 54.6 +/- 22.6 before and 62 +/- 19.0 after treatment, p=0.0112) and FEF25/75% (28.9 +/- 23.0 before and 37.5 +/- 25.5 after treatment, p=0.0053) were observed. Compared with placebo montelukast significantly improved FEV1 (p=0.0032), PEF (p=0.0298) and FEF25/75% (p=0.0091). There was no significant difference in VC, TGV, Raw and RV. Montelukast compared with placebo significantly decreased cough (p<0.0001) and wheezing (p=0.0002) score. In summary, therapy with montelukast may provide clinical benefit to patients with CF. PMID- 15757269 TI - [The prevalence of tobacco smoking among Polish Pallotins priests and seminarians of Seminary in Oltarzew]. AB - There is a lack of studies evaluating the prevalence of smoking habit among Polish priests. Due to difficulties in receiving permission from catholic authorities to perform questionnaire study we attempted to assess the prevalence of cigarette smoking among priests from one religious congregation. The aim of the study was also to analyze factors influencing smoking habits in this group. The anonymous questionnaire was sent to all Polish Pallotins and students of Pallotin Seminary in Oltarzew. Only one fourth of them--138 subjects in the age from 20 to 92 yrs (mean age 37.7 yrs)--answered the questionnaire. Among them were 72 priests aged 27-92 yrs. (mean age 50.7 yrs) and 66 students in the age 20 41 yrs (mean age 23.7 yrs.). The rate of current smokers among priests was 13.9%, including 11.1% of daily smokers and 2.8% occasional smokers. The rates of daily and occasional smokers among Seminary students were 4.5% and 6.1%, respectively. Cigarette smoking in the past was reported by 26,4% of priests--8.3% of them had smoked daily, remaining subjects had smoked occasionally. Most of responders considered the fifth commandment of the Decalogue as sufficient reason for abstaining from cigarette smoking. The majority of current smokers attempted to stop smoking, mainly because of wholesome reasons. Near a half of smoking priests and over three fourth of smoking students felt embarrassed to smoke in company of laymen. Every tenth priest and every third student hide his smoking habit from other priests. The study showed low prevalence of cigarette smoking in priests, what resulted mainly from religious reasons. PMID- 15757270 TI - [Treatment of nicotine dependence based on bupropion SR and educational support- one year observation and analysis of the reasons of relapses]. AB - Tobacco dependence is a chronic, relapsing disease causing an enormous burden of deaths. The aim of the study was a one year evaluation of the efficacy of bupropion SR supported by an educational program in the treatment of nicotine dependence and analysis of the reasons of relapses. 54 smokers were enrolled. The 12-month continuous abstinence rate was 28.3%. The most common cause of relapses in the treatment period was craving for cigarettes, in the post-treatment period stress. PMID- 15757271 TI - [Organizing pneumonia--analysis of 18 own cases]. AB - Organizing pneumonia (OP) is a rarely diagnosed disease, however the incidence ratio was estimated as 6-7/100000. Disease can occur in cryptogenic form or as a secondary reaction to various noxious agents, drugs, and ionising radiation, as a concomitant disease to infections, lympho- and myeloproliferative disorders, and connective tissue diseases. Symptoms of OP are non-specific therefore lung biopsy and histological examination are necessary for diagnosis. Eighteen cases of OP, 15 women and 3 men, aged 40 to 76 years, are presented with analysis of clinicopathological characteristic and therapeutic problems. In all cases diagnosis was confirmed by open lung biopsy. In one case radiotherapy and in one trastuzumab treatment was the cause of OP. In further 3 women antibodies against Chlamydia pneumoniae and in one--against Mycoplasma pneumoniae were found in serum. Probably Hashimoto disease was the cause of one case. In 12 patients the OP was idiopathic. Majority of patients were treated by prednisone (0.5 mg/kg). In one patient regression without any treatment was noticed and in other one- after cessation of trastuzumab. Five women were treated by clarithromycin. In 3 of them regression was observed but in other 2 corticotherapy was necessary. The observation period ranged from 1 month to 9 years, mean 34 months. PMID- 15757272 TI - [Antimycobacterial antibody level in pleural, pericardial and cerebrospinal fluid of patients with tuberculosis]. AB - The goal of the study was to evaluate IgG, IgA and IgM mediated humoral immune response against 38kDa and 16 kDa or 38kDa and LAM mycobacterial antigens in pleural, pericardial or cerebrospinal fluid from patients with tuberculosis (TB) and to compare to non-tuberculous controls (NTB). 30 cerebrospinal fluids (CSF) (16 TB pts and 14 NTB pts), 17 pericardial fluids (6 TB and 11 NTB) and 20 pleural fluids (7 TB and 13 NTB) were examined. Commercially available ELISA based assays (Pathozyme Tb complex plus, Myco G, A and M--Omega Diagnostic) were used. Tests were performed and cut off established according to manufacturer instruction. Mean IgG level against 38 + 16kDa was significantly higher in neurotuberculosis group compared to control (p<0.05). Sensitivity of the test in detecting neurotuberculosis was of 42% and specificity of 96%. Mean IgG, IgA and IgM against 38kDa + LAM level was higher in TB group compared to NTB in CSF. No difference was observed between TB and NTB group in pleural effusion. Antimycobacterial antibody levels were non-significantly increased in pericardial fluid in TB. The findings of the study indicate that TB is associated with the presence of detectable levels of antibodies in the CSF and pericardial effusion. Anti 38kDa + 16kDa IgG test can be used in combination with other diagnostic methods to increase diagnostic accuracy of neurotuberculosis. PMID- 15757273 TI - [Pulmonary fibrosis in young patients with hypersensitivity pneumonitis]. AB - Hypersensitivity pneumonitis (HP) is more common in middle-aged individuals but has been also diagnosed in patients of all ages including infants and children. Host risk factors are poorly characterized. The aim of this paper is presentation of 6 young patients in whom HP was diagnosed at the stage of lung fibrosis. There were 5 females at the age of 16-35 years and 1 male at the age of 28 years. All of them were exposed to organic dust for many years. Three of them were asthenic with scoliosis, 5 had clubbing. All patients were released from sport exercises in school due to fatigue but the diagnostic procedures were started at that time only in 2 patients. One woman had been ill from early childhood (recurrent pneumonia was recognised). In the another girl (16 years old) the spontaneous pneumothorax was the cause of the first chest X-ray examination. In the next woman (21 years old) marked dyspnea was connected with pregnancy and massive fibrotic lesions were recognised after delivery of her child. The only man was diagnosed before change of his job; he had no complaints. Two oldest women (34 and 35 years old) were diagnosed in childhood but avoidance of antigen exposure and corticotherapy were not effective. In all patients precipitins against farmer's lung and pigeon fancier's lung antigens were found. In all patients chest X-ray showed features of advanced lung fibrosis. All patients were treated with steroids without improvement. Two patients died during corticotherapy. CONCLUSIONS: 1) HP could be taken into consideration during differential diagnosis of interstitial lung diseases, 2) Cough and dyspnea on exertion could be the first symptoms of chronic HP in children also. PMID- 15757274 TI - [Lymphangioleiomyomatosis in a menopausal woman on estrogen replacement therapy]. AB - The case of lymphangioleiomyomatosis (LAM) was presented. Woman (55 years) receiving for several years estrogens with the symptoms of disease (dyspnea, nonproductive cough, chylous pleural fluid) appeared after menopause. Suggestive clinical symptoms and chest HRCT scan confirmed clinical diagnosis of LAM. Two years after discontinuation of estrogen medication her clinical condition and radiological picture of lungs improved. PMID- 15757275 TI - [Pulmonary actinomycosis]. AB - A case report of pulmonary actinomycosis is presented. The 33-years old man complained of long term pain in the thorax, coughing and subfebrile body temperature. Pleural empyema and pulmonary neoplasm were suspected. The final diagnosis was established by open thoracotomy and definitive histological examination. Partial lung resection and post operative penicillin G therapy were performed. PMID- 15757276 TI - [Exogenous lipoid pneumonia]. AB - Two cases of exogenous lipoid pneumonia are presented. The 22 years old woman was admitted to hospital because of recurrent dyspnea, cough, fever and chest pain, are changes on chest x-ray. Bronchoscopy, chest CT scan, respiratory functional test and laboratory tests were performed but the diagnosis was not established. Just before thoracoscopy patient revealed frequent intentionally choking with oil. The second patient 39 years old man was admitted because of dyspnea, cough, fever and chest pain, which appeared after accidentally choking with oil. In both cases the diagnosis was confirmed with special staining of bronchial discharge specimen. PMID- 15757277 TI - [Quality of life in chronic obstructive pulmonary disease]. PMID- 15757278 TI - [Medical aspects of staying at high altitude]. PMID- 15757279 TI - [Bupropion SR for the therapy of tobacco dependence]. PMID- 15757280 TI - [Latex allergy--Part I]. AB - Natural rubber latex (NRL), is a resin sap produced in the cells of caoutchouc plants. It is a water dispersion of cis-1,4-polisopren (caoutchouc)--35%, stabilized with little amounts of proteins, sugar, alcohol, fatty acids and salts. The concentration of all solid substances is about 40%, the rest is water. Immunogenicity of latex depends on the proteins it contains. For many years we read in medical papers about the cases of contact urticaria, asthma, rhinitis, and anaphylaxis after contacting with latex products. It turns out that medical staff is the group of high occupational risk, because of exposure to gloves and other latex products. It is connected with the fact of high gloves usage caused by the danger of virus infections: HIV, HBV, HCV. Latex allergy is one of the reasons of dramatic complications after surgical operations. People who are allergic to latex may have cross reactions to allergens not connected with occupational environment. These are: food and houseplants (Ficus benjamina). The frequency of latex allergy is about 0.1% of the population. In the groups of high risk the frequency rises sharply. It is 17% among medical staff and it reaches 60% among children with spina bifida. PMID- 15757281 TI - [Latex allergy--Part II]. AB - Diagnosis of latex allergy consists of four methods: anamnesis, skin tests, laboratory examinations and specific allergen challenge. Anamnesis is the source of information on the symptoms and reactions of IgE-dependent allergy, different accompanying factors and atopy diseases. The recognition of latex allergy is possible with positive SPT and/or sIgE (latex). Specific allergen challenge is performed in situation with positive anamnesis, negative SPT and negative sIgE (latex). Latex allergy treatment is not different than other allergic diseases. Specific immunotherapy is new methods in these cases. Because of absence of standardized latex allergens, this method is still on experimental issue. The inconsistent results do not allow the recommendation of immunotherapy in everyday practice. The most important prophylactic activity in case of latex allergy is to reduce the exposure to latex among the medical staff and the patients. PMID- 15757282 TI - [Activity of Jakub hr. Potocki Foundation]. PMID- 15757283 TI - [Functional nasal surgery in the treatment of obstructive sleep apnea]. AB - The study included 22 males with significant decrease in nasal patency, at age of 44+/-7 yrs with body mass index 28.9+/-3.8 kg/m2, diagnosed with obstructive sleep apnea syndrome (OSAS) by polysomnography. All patients underwent functional, corrective nasal surgery. In one patient an infection in the wound occurred. Postoperatively 19 (86%) patients reported significant subjective improvement. With regard to polysomnography, one patient was cured and in another one a decrease of AHI to more than 50% of baseline was found. In 6 (27.3%) patients AHI rose from 33.2+/-13 to 53.6+/-21.2. CONCLUSION: Nasal surgery in OSAS shows limited effectiveness. Because of multilevel decrease in airway patency, some of the patients may need a step-wise approach to surgical treatment. PMID- 15757284 TI - [Serum level of leptin and neuropeptide Y in children with mild asthma]. AB - Leptin (LEP) and neuropeptide Y (NPY) are involved in the maintenance of energy balance and create regulatory loops on central and peripheral stage between neuropeptides and hormones, additionally regulated by other physiological stimuli. According to data confirming tendency to overweight and obesity in asthmatic children we have examined the influence of mild asthma on neurohormonal balance. 43 children, aged 7-17 years, including 27 steroid naive mild asthmatic children aged (mean+/-SD) 12.3+/-2.6 years and 16 age matched healthy children participated in the study. Serum LEP and NPY levels were measured radioimmunologically (RIA). Serum leptin level in asthmatic children was 2.84+/ 2.1 ng/ml and did not differ to that of healthy children -3.49+/-1.65 ng/ml, both in boys (p=0.85) and girls (0.49). Similarly, we did not observe any differences between NPY levels in asthmatic (113.5+/-31.1 micromol/ml) and healthy (98.5+/ 21.9 micromol/ml (p=0.17)), irrespective to sex. Significant correlations between leptin levels and body mass index (BMI) in asthmatics (r=0.62, p=0.01) and healthy children (r=0.56, p=0.02) were observed. We conclude, that mild asthma in children seems not to affect neurohormonal regulation of energy balance. PMID- 15757285 TI - [Antieosinophil action of IL-12 in human polyp culture]. AB - RATIONALE: Tissue eosinophilia is an important pathophysiological issue in bronchial asthma and nasal polyps. Its magnitude is regulated by several mechanisms, including selective migration of eosinophils to peripheral tissues and prolongation of survival. A key role in this processes is assigned to Th2 originating cytokines: IL-3, IL-5, GM-CSF. It has been previously demonstrated that IL-12 diminishes tissue eosinophilia in an animal model, and Hofstra showed that IL-12 together with IL-18 prevents allergen-induced increase bronchial hyperresponsiveness, BAL eosinophilia and the development of allergen-specific Th2 cells. METHODS: Nasal polyps were obtained during routine surgery and were cultured in fragments of approximately 30 mg for 2, 6, and 15 days in RPMI 1640 in the absence or presence of IL-12. Afterwards a dose-dependency was tested at day 2 of culture. Polyp tissue from cultures was than processed to slides, stained with Giemsa and cells were counted in light microscopy (400x). RESULTS: Eosinophils represented 62.8+/-21.3% of residing cells in nasal polyps at the day 0. IL-12 (1 microg/ml) caused a significant time-dependent decrease in the percentage of Eos after 2 and 6 days. The effect of IL-12 at day 2 was concentration-dependent: control, 28.2+/-2.9; at 10 ng/ml, 13.9+/-6.4 (n=4, p<0.05); at 100 ng/ml, 11.6+/-2.1 (p<0.01); at 1 microg/ml, 7.5+/-1.5 (p<0.005). CONCLUSION: IL-12 acts as potent topical antieosinophilic agent. Its action can be seen in a cultured polyp environment. It is visible already after two days and is concentration-dependent. Further study is needed to elucidate tissue mechanisms of this action. PMID- 15757286 TI - [Autoimmune thyroid diseases in patients with primary pulmonary hypertension]. AB - Abnormal metabolism due to thyroid disorders could cause the additional significant right ventricular (RV) overload in patients with primary pulmonary hypertension (PPH). The purpose of this study was the prospective determination of autoimmune thyroid status in PPH patients. MATERIAL: 45 patients (33 women and 12 men, mean age 38 years+/-13) with moderate and severe PPH (mean pulmonary artery pressure 61 mmHg+/-18 mmHg). PPH was diagnosed when other reason of pulmonary hypertension had been excluded especially thromboembolic, venous and hypoxemic. METHODS: Titer of antibodies to thyroglobulin and thyroid peroxidase were measured. Thyroid gland dysfunction was determined by biochemical measurements of thyrotropin and free thyroxine; thyroid ultrasonographic examination was also performed. The prevalence of thyroid autoantibodies is estimated at 13% in general population. RESULTS. In 14 patients (31%) increased titer of thyroid antibodies was found in our group. Hyperthyroidism was recognized in 2 patients and in 3 cases we diagnosed hypothyroidism with high antibodies level requiring treatment. Ultrasonographic abnormalities were revealed in 21 patients (nodular changes--11 patients, fibrosis--3 patients, chronic inflammation symptoms--7). Ultrasonographic abnormalities were present in all patients with detectable antithyroid antibodies. We haven't confirmed more frequent prevalence of hormonal disorders in patient treated with prostacyclin's analogues. CONCLUSION: The prevalence of autoimmune thyroid diseases is about 3 times higher in PPH patients than in general population. The treatment of patients with clinically evident hyper- or hypothyroidism is necessary. The monitoring of clinically silent disorders seems to be recommended especially prior to lung transplantation qualification. PMID- 15757287 TI - [Adenocarcinoma of the lung with metastases to the pituitary and both adrenal glands]. AB - A case of lung adenocarcinoma with metastases in both adrenals and pituitary gland in 52-year old patient is presented. Adrenal glands metastases were diagnosed on a base of CT scans. Pituitary metastases caused gradual loss of vision. MR imaging showed a lesion in the sella and suprasellar areas and suggested meningioma. Right frontal craniotomy with the partial removal of the tumor was performed. Three days later the patient died. Histopathological results of the pituitary masses revealed adenocarcinoma. PMID- 15757288 TI - [Tuberculosis of the tongue in a patient with disseminated pulmonary tuberculosis]. AB - A 51-year-old man, heavy cigarette smoker, a homeless alcoholic, with disseminated lesions in lungs and with ulceration and infiltration of the tongue is presented. Treatment with antibiotics was ineffective. He was admitted to the otolaryngological department because of suspicion of the tongue cancer. The histological examination of the tongue biopsy revealed tuberculous granuloma. In the pulmonological department, on admission the patient was cachectic, with massive oedema and ulceration of the tongue, and enlargement of the cervical lymph nodes. He was fed through the gastric tube. He had severe pain of the tongue demanding treatment with opiates analgesics. Chest x-ray revealed disseminated lesions in lungs. Antituberculous therapy was administered because of suspicion of tuberculosis of the tongue and lungs. During the treatment clinical improvement was observed. Tubercule bacilli were grown in the sputum culture after 6 weeks of observation. After 8 weeks of antituberculous therapy regression of lung lesions and healing of the tongue were observed. The patient continued treatment for 6 months in the Lung Disease Outpatient Clinic. PMID- 15757289 TI - [Solitary fibrous tumor of the pleura in 75-year old woman]. AB - A 75-year-old asymptomatic woman with stable coronary heart disease presented tumor in lower left lobe on routine chest radiograph. A CT scan showed a large sharply delineated mass at this site (84 x 52 x 90 mm). There were no signs of infiltration, no abnormalities were seen in mediastinal structures and on the right side. The pedunculated tumor was resected during left thoracotomy (posterolateral incision). Histological examination revealed spindle-like cells and rich collagen net. Mitoses and necrosis were absent. Final diagnosis was: solitary fibrous tumor of the visceral pleura. During 7-year follow-up recurrence was not observed. PMID- 15757290 TI - [Role of immunoglobulin E in bronchial hyperreactivity development]. PMID- 15757291 TI - [Pulmonary-renal syndrome]. PMID- 15757292 TI - [The role of leukotrienes in inflammatory process in cystic fibrosis]. PMID- 15757293 TI - [Quality of life in children with asthma]. PMID- 15757294 TI - [Chronic cor pulmonale--40 years later]. PMID- 15757295 TI - [Does apoptosis play a role in the pathogenesis of chronic obstructive pulmonary disease?]. PMID- 15757296 TI - [Therapeutic possibilities and clinical effects of IgE level modulation in allergic diseases]. PMID- 15757297 TI - [Timofei Petrovich Krasnobaev (1865-1952)]. PMID- 15757298 TI - [The endoscopic and morphological diagnostics of gastroesophageal reflux]. AB - The experiences with endoscopic examinations of 140 patients with gastroesophageal reflux followed by the development of reflux-esophagitis (RE) of different degree are presented. The authors describe the complicated and uncomplicated forms of RE. The uncomplicated forms include RE with unimpaired integrity of the epithelium, erosive RE, erosive-ulcerous RE. The complicated forms include peptic ulcer of the esophagus, peptic stricture and Barrett esophagus. The degree of RE was found to depend on the character and degree of impairment of the obturating function of the cardia. The morphological examination of the esophagus mucosa was performed in 35 patients. Morphological heterogeneity of the so called "catarrhal RE" was shown that makes the expedience of using this term in clinical practice doubtful. Exact endoscopic and morphological criteria of the differential diagnostics of erosive and erosive ulcerous RE, erosive-ulcerous RE and peptic ulcer of the esophagus are described. PMID- 15757299 TI - [Surgical aspects of periodic disease abdominal paroxysms]. AB - Thorough investigation of abdomenalgia symptoms in 220 patients allowed to reveal a number of peculiarities of the periodic disease (PD) promoting the correct diagnosis. The case history data, pain localization, fever and great potentialities of diagnostic laparoscopy carried on during the acute paroxysm of the abdominal form of PD are of great significance. The differential diagnosis of PD acute paroxysm and urgent pathology of the organs of the abdominal cavity are considered by the author to be possible when using diagnostic laparoscopy in difficult situations. PMID- 15757300 TI - [The peculiarities of lymph outflow from the stomach and characteristics of groups of regional lymph nodes]. PMID- 15757301 TI - [Means of reducing blood loss and volume of the used transfused media in operative treatment of burn lesions of the liver]. AB - An experience with surgical treatment of 77 patients with focal lesions of the liver is described. The patients were divided into two groups. In the main group (42 patients) the treatment-and-prophylactic method was used including acute isovolemic and hypervolemic hemodilution, preliminary preparation of autoblood, isolation and ligation of the vascular-secretory elements, the application for local hemostasis with Takhokomb of "Tissucol", gelatinous sponge with gentamycin. In the group of comparison the compression of the hepatoduodenal ligament, isolation of the vascular-secretory elements by digitoclasia method, suturing the liver stump with polysorb were used in resection of the liver. The strategy used in the main group allowed to reduce the volume of blood loss, to lessen the number of doses of the transfused donor blood, to diminish the number of postoperative complications by 30.5%. The used complex is effective, simple and is not expensive. PMID- 15757302 TI - [The sphincter-related biliodigestive anastomoses in anomalies of bile ducts]. AB - The authors made an analysis of anomalies of the bile ducts with special reference to anatomical peculiarities and iatrogenic lesions of the bile ducts. The anomalies of bile ducts were found in 2.07% of 676 patients operated on for complicated cholelithiasis. Primary operative interventions and reoperations in the patients were completed with the formation of the constriction-biliodigestive anastomoses of different levels having antireflux functions. The authors describe the operation technique and results of the treatment of bile duct obstruction. PMID- 15757303 TI - [Ileoascendoanastomosis]. AB - A complex anatomical investigation of the extra- and intra-organic vessels of the ascending colon has shown that the formation of ileoascendoanastomosis is possible and expedient without a preliminary intraoperative assessment of the vascular bed. The indications to such operation are developed. The extirpation of the blind gut and resection of the ascending colon with applying the anastomosis in question was performed on 6 patients with carcinoma of the blind gut and ascending colon. The results of operations were good. PMID- 15757304 TI - [Decision for the method of surgical treatment of acute destructive pancreatitis]. AB - The work presents results of treatment of 375 patients with acute pancreatitis, 93 patients with destructive pancreatitis were operated upon. Minimally invasive and traditional technologies were used. Postoperative lethality was 6.5%. The method of lumboscopic extraperitoneal drainage of the destruction focus under the control of laparoscopy and USI is described. The percutaneous paracenthetic draining interventions under the control of USI and endovideosurgical technologies were shown to be highly effective in treatment of patients with acute destructive pancreatitis and its complications. PMID- 15757305 TI - [Potentialities of hemodilution and photocorrection of blood in treatment of patients with obliterating atherosclerosis of lower extremities]. AB - The state of microcirculation in the ischemized extremity fingers was studied in 89 patients with obliterating atherosclerosis of the lower extremity vessels treated by conservative rheological therapy with rheopolyglucin and intravascular laser irradiation of blood in order to define their application alone, or in combination or in succession. The work has shown evident effectiveness of their using alone and in succession, while a combined use of the methods in question was found to be not expedient. The successive use of these methods can be considered expedient when monotherapy had proved to be helpless. PMID- 15757306 TI - [The experimental grounds and clinical application of domestic endoprosthesis "Esfil" for plasty of the abdominal wall]. AB - An investigation of physico-mechanical properties and a histological examination of meshed endoprostheses "Esfil" ("Lintex" St.Petersburg) and nets "Prolene" ("Ethicon") implanted into the abdominal wall of 66 Chinchilla rabbits was carried out under experimental conditions. The comparative experimental investigation has shown that both materials possess sufficient physico-mechanical properties to resist intra-abdominal pressure and when implanted they cause an inconsiderable inflammatory reaction followed by the formation of a solid connective tissue capsule. The endoprosthesis "Esfil" was used for plasty of the abdominal wall in 51 patients with postoperative ventral hernias. The endoprosthesis "Esfil" was found to meet all the requirements made to endoprostheses for plasty of the abdominal wall. Its use in clinical practice allows the restoration of quality of life of such patients. PMID- 15757307 TI - [Pathogenetical features of the acute period of traumatic disease. Traumatic shock as a special manifestation of the acute period]. AB - Clinico-pathogenetical features of the acute period of trauma disease due to a severe combined injury were studied in 644 patients. An analysis of clinical indices was made in 428 of them, clinico-laboratory indices--in 216 patients. The results have shown that the main form of the acute period of the trauma disease are traumatic shock (62.8%), traumatic coma (18.3%), acute respiratory insufficiency (5.7%). Dramatically decreased delivery and consumption of oxygen lower than the metabolic requirements of the tissues accompany the traumatic shock which results in the suppression of tissue metabolism. Pronounced endotoxicosis developing along with the traumatic shock results not from the degree of injured tissues only, but from the centralization of blood circulation. It is also supported by the insufficient detoxicating function of the kidneys. A severe degree of the traumatic shock causes disseminated intravascular coagulation (DIC) of the II-III degree. After helping out from shock the patient has normocoagulation or the I degree DIC. Traumatic shock with marked hypotension and metabolic acidosis is a significant factor of high risk of the development of acute lesion of the lungs. PMID- 15757309 TI - [Combined mini-invasive operations in gastroduodenal ulcers]. AB - The authors' experiences show that at the present-day stage of the development of mini-invasive surgery the combined method of operating consisting of laparoscopic and mini-laparotomic steps is a perspective alternative. In addition, the authors note high efficiency of combined operations having all advantages of mini invasive surgical technologies which was confirmed by investigations of the effects of access on microcirculation of the peritoneum in different surgical accesses used in their clinic. PMID- 15757308 TI - [Oxygen transport and hemodynamic reactions in ischemic heart disease patients when reserving and reinfusing massive volumes of autoblood]. AB - The prospective randomized investigation was carried out in 61 patients with ischemic heart disease who were subjected to planned operations of coronary shunting under conditions of extracorporeal circulation. In the first group of patients autoblood as much as 24% of the calculated circulating blood volume (CBV) was reserved at the stage of isolation of the mammary artery, before systemic heparinization. The volume of the reserved blood of the patients of the second group was about 12% of CBV. It was shown that the reserving and reinfusion of massive (about 24% of CBV) volumes of autoblood in surgical treatment of ischemic heart disease under conditions of extracorporeal circulation failed to cause a substantial change of indices of central hemodynamics as compared to the reserving and reinfusion of autoblood in volume of about 12% of CBV. PMID- 15757310 TI - [The clinical-morphological estimation of the results of laser osteoperforation in treatment of chronic osteomyelitis]. AB - The method of laser osteoperforation was approved in 135 patients with chronic osteomyelitis. The diode laser with wave length 970 mm and peak power up to 30 Wt was used. Energy was delivered by the percutaneous contact method with a quartz light guide of 0.4 mm diameter. All the patients had a distinct positive reaction in the nearest days after operation. The fistulas were closed within 4-10 days. Stable remission was noted during 2-5 years by objective indices (X-ray, fluometry, densitometry, morphological data). Our experiences with clinical use of laser osteoperforation show its high efficiency in treatment of chronic osteomyelitis. The method is minimally invasive and allows to considerably shorten the terms of treatment and rehabilitation. PMID- 15757311 TI - [An experience with stenting the esophagus in patients with inoperable cancer]. AB - An experience with treatment of 21 patients with inoperable cancer of the esophagus allowed determination of indications and contraindications, development of the method and technique of endoscopic implantation of a domestic meshed self expanding stent. The complications resulting from the process of mounting and the stent itself are analyzed. The methods of eliminating the complications are shown. The immediate and nearest results of endoprosthesing are assessed. Recommendations are given concerned with the cases of ingrowth of tumor into the stent. PMID- 15757312 TI - [Effectiveness of collagen plasty in complex treatment of localized pyopneumothorax]. AB - The work is based on an analysis of results of treatment of 25 patients with localized pleuro-pulmonary cavities by the methods of minor surgery. During abscessoscopy collagen was placed on the wound surface, the surface was filled with fibronectin and followed by active aspiration within 12-24 hours. In 20 (80%) patients the lung was expanded earlier. Five patients had no effect. PMID- 15757313 TI - [Laparoscopic splenectomy through a lateral access]. AB - The investigation included 35 patients with diseases of blood and spleen pathology (30) and wounds and traumas of the abdomen with injuries of that organ (5), treated by laparoscopic splenectomy through a lateral access. The technique and stages of the operation and advantages of the lateral access are described. The laparoscopic splenectomy was shown to be possible in 90% of cases if patients were selected with special reference to the main disease, size and degree of pathological alterations in the spleen. An adequate approach to selections of patients with injuries of the organ allowed to make operations in more than a third of the patients and casualties. When using the lateral access laparoscopic splenectomy is thought to be more convenient, gives better visualization of the anatomical structures and reduces risk of injury of adjacent organs, thus resulting in less frequency of complications and conversions. PMID- 15757314 TI - [Strategy of treatment for bleeding of portal genesis]. AB - Based on a comparative analysis of results of the surgical treatment of bleedings from varicose veins of the liver and stomach, the authors recommend differential decision for the variants of surgical interventions at different periods from the moment of the appearance of bleeding and depending on the severity of impairments of the liver functions. PMID- 15757315 TI - [Can we agree? (Apropos of the article by A. D. Aslanov et al. "Comparative assessment of different methods of treatment of critical ischemia of lower extremity in elderly and senile patients".--Vest. Chir.--2003, No 4.-- P. 83)]. PMID- 15757316 TI - [Specialized medical service for polytrauma in a large city]. AB - On the basis of studying of trauma epidemiolgy in a large city, principles of optimal organization of specialized medical service for patients with polytrauma have been developed. Urgent specialized service and further treatment of patients with this pathology should be fulfilled in a special department organized on the basis of a multi-profile hospital. The center of this department represents a specialized unit for patients with multiple and combined traumas supplied with all necessary specialists and means required for the management of this difficult group of patients. PMID- 15757318 TI - Coastal waters need action. PMID- 15757317 TI - [Operative accesses in breast tumors with Pancoast's syndrome]. PMID- 15757319 TI - Canada bans fluoropolymer stain repellents. PMID- 15757320 TI - Funding woes eroding stream gage network. PMID- 15757321 TI - Pollutants persist in drinking water. PMID- 15757322 TI - Cleaning up school bus emissions. PMID- 15757323 TI - Mine tailings soak up greenhouse gas. PMID- 15757324 TI - Taking the mass out of transit. PMID- 15757325 TI - Recovery of acidified European surface waters. PMID- 15757326 TI - Framing the elusive concept of sustainability: a sustainability hierarchy. AB - Usage of the word "sustainability" is widespread and incorporates a plethora of meanings. After reviewing four extant sustainability frameworks, we propose a Sustainability Hierarchy to structure a broad array of issues that have been associated with sustainability. These issues vary widely in their urgency, severity and uncertainty of consequences, and temporal and spatial dimensions. It categorizes actions some view as unsustainable based on their direct or indirect potential to (i) endanger the survival of humans; (ii) impair human health, (iii) cause species extinction or violate human rights; or (iv) reduce quality of life or have consequences that are inconsistent with other values, beliefs, or aesthetic preferences. Effects considered include impediments to the ecosystem functions that support human life, human health, and species viability. This paper argues that for sustainability to become a more meaningful concept, the many worthy issues in the fourth category (values, beliefs, and aesthetic preferences) should not be considered sustainability concerns. Implications for companies, policy makers, and scientists are discussed. PMID- 15757327 TI - Choosing chemicals for precautionary regulation: a filter series approach. AB - The present case study develops and applies a systematic approach to the precautionary pre-screening of xenobiotic organic chemicals with respectto large scale environmental threats. It starts from scenarios for uncontrollable harm and identifies conditions for their occurrence that then are related to a set of amplifying factors, such as characteristic isotropic spatial range p. The amplifying factors related to a particular scenario are combined in a pre screening filter. It is the amplifying factors that can transform a potential local damage into a large-scale threat. Controlling the amplifying factors means controlling the scope and range of the potential for damage. The threshold levels for the amplifying factors of each filter are fixed through recourse to historical and present-day reference chemicals so as to filter out as many as possible of the currently regulated environmental chemicals and to allow the economically important compounds that pose no large-scale environmental concern. The totality of filters, with each filter corresponding to a particular threat scenario, provides the filter series to be used in precautionary regulation. As a demonstration, the filter series is then applied to a group of nonreferential chemicals. The case study suggests that the filter series approach may serve as a starting point for precautionary assessment as a scientific method of its own. PMID- 15757328 TI - Persistent organic pollutants in two dolphin species with focus on toxaphene and polybrominated diphenyl ethers. AB - Assessing trends of persistent organic pollutants (POPs) in marine mammals is difficult due to age, gender, and metabolism influences on accumulation. To help elucidate these effects in dolphins, POP concentrations were determined in the Atlantic white-sided dolphin, Lagenorhynchus acutus, a pelagic delphinid inhabiting North Atlantic waters, and in the rough-toothed dolphin, Steno bredanensis, a pelagic delphinid inhabiting tropical and subtropical waters. The specific objectives of this study were to determine baseline POP concentrations in L. acutus and S. bredanensis blubber samples and to examine the effects of age, gender, and metabolism on POP concentrations in dolphin blubber. Focus was aimed at contaminants of emerging concern, specifically, toxaphene and polybrominated diphenyl ethers (PBDEs). Samples collected from L. acutus (n = 47) stranding events in Massachusetts (1993--2000) and S. bredanensis samples (n = 15) were analyzed for PCBs, toxaphene, and other organic pesticides by gas chromatography/mass spectrometry (GC/MS). Age and gender influences were similar between the two species, with adult females having significantly lower POP concentrations as compared to adult males and juveniles. Mean sigma toxaphene concentrations were highest in juvenile L. acutus, 13.0 (6.7) microg/g wet mass (1 SD), and lowest in adult female S. bredanensis, 1.49 (1.4) microg/g wet mass. sigma PBDE (sum of congeners 47, 99, 100, 153, and 154) concentrations were highest in juvenile L. acutus, 2.41 (1.2) microg/g wet mass, and lowest in adult female S. bredanensis, 0.51 (0.6) microg/g wet mass. POP concentrations did not significantly differ between adult males and juveniles, suggesting metabolism of congeners and/or dilution with growth. PBDE concentrations in juvenile white sided dolphins were not significantly related to collection year, suggesting that there may be a lag period for higher concentrations to be detected in pelagic marine mammals such as L. acutus or that concentrations have already peaked in this species prior to the first collection in 1993. PMID- 15757329 TI - Size and composition of airborne particles from pavement wear, tires, and traction sanding. AB - Mineral matter is an important component of airborne particles in urban areas. In northern cities of the world, mineral matter dominates PM10 during spring because of enhanced road abrasion caused by the use of antiskid methods, including studded tires and traction sanding. In this study, factors that affect formation of abrasion components of springtime road dust were assessed. Effects of traction sanding and tires on concentrations, mass size distribution, and composition of the particles were studied in a test facility. Lowest particle concentrations were observed in tests without traction sanding. The concentrations increased when traction sand was introduced and continued to increase as a function of the amount of aggregate dispersed. Emissions were additionally affected by type of tire, properties of traction sand aggregate, and driving speed. Aggregates with high fragmentation resistance and coarse grain size distribution had the lowest emissions. Over 90% of PM10 was mineral particles. Mineralogy of the dust and source apportionment showed that they originated from both traction sand and pavement aggregates. The remaining portion was mostly carbonaceous and originated from tires and road bitumen. Mass size distributions were dominated by coarse particles. Contribution of fine and submicron size ranges were approximately 15 and 10% in PM10, respectively. PMID- 15757330 TI - Characterization of organic compounds collected during southeastern aerosol and visibility study: water-soluble organic species. AB - As part of the Southeastern Aerosol and Visibility Study (SEAVS), water-soluble organic species (WSOS) in fine aerosols collected from July 15 to August 25, 1995, at the Great Smoky Mountain National Park, Tennessee (USA), were chemically classified into seven groups, with concentrations ranging from around 1 to >200 ng/m3. Dicarboxylic acids represented the dominant identified compound class, and succinic acid was the most abundant dicarboxylic acid. The trends in data suggest that most WSOS collected in the SEAVS samples were mainly generated from secondary photochemical reactions, especially during the first (cleaner) half of the sampling campaign. High relative humidity at the sampling site resulted in substantial water uptake by the aerosols, which may have enhanced the levels of succinic acid by reducing its rate of photooxidation. Concurrent trends in malic and malonic acid concentrations suggest these were generated from the oxidation of succinic acid. Consistent with the conversion of 3-hydroxypropanoic acid to malonic acid, it appears that 4-hydroxybutanoic acid served as a major precursor contributing to high levels of succinic acid in the daytime. Nocturnal WSOS generally followed the trend of diurnal WSOS, but they exhibited different chemical compositions and lower concentrations, unlike what has been reported for an urban site. A nocturnal-to-diurnal ratio of succinic acid larger than 0.25 may indicate an atmosphere dominated by photochemical reactions, rather than by primary emissions. PMID- 15757331 TI - Declining threshold for hypoxia in the Gulf of Mexico. AB - The northwestern Gulf of Mexico shelf has been nicknamed "The Dead Zone" due to annual summertime (May-September) bottom-water hypoxia (dissolved oxygen < or =2 mg L(-1)) that can be extensive (>20 000 km2) and last for several months. Hypoxia has been attributed to eutrophication caused by increasing nitrogen loads, although directly linking hypoxia to nitrogen is difficult. While the areal extent of hypoxia has been shown to increase with Mississippi River flow, it is unclear whether this increase results from enhanced vertical water-column stratification or from eutrophication caused by river-borne nutrients. Disentangling the relative contributions of eutrophication versus stratification has important management consequences. Our analysis indicates that the top:bottom salinity difference is an important predictor of hypoxia, exhibiting a threshold, where the probability of hypoxia increases rapidly, at approximately 4.1 ppt. Using a Bayesian change-point model, we show that this stratification threshold decreased from 1982 to 2002, indicating the degree of stratification needed to induce hypoxia has gone down. Although this declining threshold does not link hypoxia and nitrogen, it does implicate a long-term factor transcending yearly flow-induced stratification differences. Concurrently, we show that surface temperature increased, while surface dissolved oxygen decreased, suggesting that factors in addition to nitrogen may be influencing the incidence of hypoxia in the bottom water. PMID- 15757332 TI - Hexachlorocyclohexanes and endosulfans in urban, rural, and high altitude air samples in the Fraser Valley, British Columbia: evidence for trans-Pacific transport. AB - High-volume air samples collected over the period Aug. 14-30, 2001, in the Lower Fraser Valley, BC, Canada, were used to assess urban/rural differences of organochlorine pesticides (OCPs) for ground level samples and to attempt to directly measure events of trans-Pacific inputs through the mid-troposphere. Hexachlorocyclohexanes (alpha- and gamma-isomers; 2-25 pg m(-3)) and endosulfan1 and -2 (5-150 pg m(-3)) were detected in all ground level samples. Seven air samples were collected during mid-troposphere flights (approximately 4400 m altitude) over the Lower Fraser Valley. These flights occurred concurrently with ground level sampling. Trajectory analysis identified three events of substantial mid-troposphere, trans-Pacific flow where 10-day back trajectories stemmed from potential source regions in Asia. These events were also characterized by higher air concentrations of alpha-HCH at 4400 m as compared to the ground level stations. This represents the first event-based, aircraft measurement of advection inputs of OCPs in the mid-troposphere of the west coast of North America. PMID- 15757333 TI - Biodegradability, DBP formation, and membrane fouling potential of natural organic matter: characterization and controllability. AB - Various natural organic matter (NOM) constituents were evaluated in terms of their biodegradability, disinfection byproduct (DBP) formation potentials, and membrane fouling. The biodegradability of NOM was evaluated with respect to biodegradable dissolved organic carbon (BDOC) and its inhibition control. NOM was divided into (i) colloidal and noncolloidal NOM, using a dialysis membrane with a molecular weight cutoff of 3500 Da and (ii) hydrophobic, transphilic, and hydrophilic NOM constituents, using XAD-8/4 resins. The colloidal, and noncolloidal hydrophilic, NOM were identified as being more problematic than the other components, exhibiting relatively higher biodegradability and reactivity toward DBP formation potential. A higher biodegradability especially can provide a high risk of membrane biofouling, if a membrane is fouled by highly biodegradable NOM. Colloidal, and noncolloidal hydrophilic, NOM constituents were also shown as major foulants of negatively charged membranes due to their high neutral fractions. Filter adsorber (F/A) types of activated carbons were evaluated in terms of removals of NOM, DBP formation potential, and BDOC and were compared to conventional processes and a nanofiltration membrane. The F/A process exhibited a comparatively good efficiency, especially in DBP and BDOC control, but was not so good at removing NOM. This suggests that F/A could potentially be combined with a membrane process to minimize the DBP formation potential and bio /organic-fouling (i.e., F/A process as a pretreatment for a membrane process). PMID- 15757334 TI - Temperature dependence of atmospheric PCB concentrations. AB - An analysis of data from the U.S. Integrated Atmospheric Deposition Network (IADN) sites near the Great Lakes and a review of the literature shows that the temperature dependence of atmospheric PCB concentrations cannot be used to distinguish sites dominated by long-distance transport from those with local sources. We observe that calculations based on data sets with only approximately 25 measurements over a period of 1 year are unreliable indicators of the long term temperature dependence at a given location, that temperature independence occurs at temperatures below freezing, and that low PCB concentrations can bias analyses toward a weaker temperature dependence. After accounting for these factors, a similar temperature dependence is observed at all IADN sites, and this temperature dependence increases for PCBs with increasing chlorination. Although previously described models do fit the data, the resulting calculated constants are not meaningful. Instead, a simple new model in which temperatures < 273 K are set equal to 273 K, and the Clausius-Clapeyron equation is used, can account for observed temperature dependence phenomena. PMID- 15757336 TI - Influence of amine buffers on carbon tetrachloride reductive dechlorination by the iron oxide magnetite. AB - The influence of amine buffers on carbon tetrachloride (CCl4) reductive dechlorination by the iron oxide magnetite (FeIIFeIII2O4) was examined in batch reactors. A baseline was provided by monitoring the reaction in a magnetite suspension containing NaCl as a background electrolyte at pH 8.9. The baseline reaction rate constant was measured at 7.1 x 10(-5)+/-6.3 x 10(-6) L m(-2) h(-1). Carbon monoxide (CO) was the dominant reaction product at 82% followed by chloroform (CHCl3) at 5.2%. In the presence of 0.01 M tris (deuteroxymethyl)aminomethane (TRISd), the reaction rate constant nearly tripled to 2.1 x 10(-4)+/-6.5 x 10(-6) L m(-2) h(-1) but only increased the CHCl3 yield to 11% and did not cause any statistically significant changes to the CO yield. Reactions in the presence of triethylammonium (TEAd) (0.01 M) increased the rate constant by 17% to 8.6 x 10(-5)+/-8.1 x 10(-6) L m(-2) h(-1) but only increased the CHCl3 yield to 8.8% while leaving the CO yield unchanged. The same concentration of N,N,N',N'-tetraethylethylenediamine (TEEN) increased the reaction rate constant by 18% to 8.7 x 10(-5)+/-4.8 x 10(-6) L m(-2) h(-1) but enhanced the CHCl3 yield to 34% at the expense of the CO yield that dropped to 35%. Previous work has shown that CHCl3 can be generated either through hydrogen abstraction by a trichloromethyl radical (radical CCl3), or through proton abstraction by the trichlorocarbanion (-:CCl3). These two possible hydrogenolysis pathways were examined in the presence of deuterated buffers. Deuterium tracking experiments revealed that proton abstraction by the trichlorocarbanion was the dominant hydrogenolysis mechanism in the magnetite-buffered TRISd and TEAd systems. The only buffer that had minimal influence on both the reaction rate and product distribution was TEAd. These results indicate that buffers should be prescreened and demonstrated to have minimal impact on reaction rates and product distributions prior to use. Alternatively, it may be preferable, to utilize the buffer capacity of the solids to avoid organic buffer interactions entirely. PMID- 15757335 TI - Biodegradation of 2,4,6-tribromophenol during transport in fractured chalk. AB - The effect of physicochemical conditions (residence time, oxygen concentrations, and chalk characteristics) on the biodegradation of 2,4,6-tribromophenol (TBP) during transport was investigated in low-permeability fractured-chalk cores. Long term (approximately 600 d) biodegradation experiments were conducted in two cores (approximately 21 cm diameter, 31 and 44 cm long, respectively), intersected by a natural fracture. TBP was used as a model contaminant and as the sole carbon source for aerobic microbial activity. Bacterial isolates were recovered and identified by both Biolog identification kit and 16S rDNA sequences from batch enrichment cultures. One of the strains, with 98% similarity (based on the 16S rDNA data) to Achromobacter xylosoxidans, was shown to have the ability to degrade TBP in the presence of chalk. The decrease in TBP concentration along the fracture due to biodegradation was not affected by reducing the residence time from 49 to 8 min. In contrast, adding oxygen to the water at the inlet and increasing the flow rates improved TBP removal. Although the matrix pore-size distribution limits microbial activity to the fracture void, the chalk appears to provide an excellent environment for biodegradation activity. Approximately 90% of TBP removal occurred within 10 cm of the TBP source, indicating that in-situ bioremediation can be used to remove organic contaminants in low-permeability fractured rocks if nutrient-delivery pathways within the aquifer are secured. PMID- 15757337 TI - Importance of unburned coal carbon, black carbon, and amorphous organic carbon to phenanthrene sorption in sediments. AB - The aim of this paper was to estimate the contribution to total phenanthrene sorption from unburned coal and black carbon (BC; soot and charcoal) in sediment. We determined sorption isotherms for five Argonne Premium Coal standards over a wide concentration interval (0.01-10 000 ng/L). The coals showed strong and nonlinear sorption (carbon-normalized K(F) = 5.41-5.96; nF = 0.68-0.82). Coal sorption appeared to become more nonlinear with increasing coal maturity. The coal's specific surface area appeared to influence K(F). On the basis of the current coal sorption observations combined with earlier petrographic analyses and BC sorption experiments, we calculated for one particular sediment that coal, BC, and "other" OC were all important to PHE sorption in the environmentally relevant nanogram per liter range. This indicates that it is important to consider strong sorption to coal in the risk assessment of coal-impacted geosorbents (e.g., river beds) where coal is mined/shipped and manufactured gas plant sites. PMID- 15757338 TI - Quantification of abiotic reaction rates in mine tailings: evaluation of treatment methods for eliminating iron- and sulfur-oxidizing bacteria. AB - Effective treatment techniques for eliminating iron-oxidizing (IOB) and sulfur oxidizing bacteria (SOB) are required for the comparison of abiotic and microbial sulfide oxidation rates and mechanisms in mine tailings. This study evaluates the effect of autoclaving, repeated heating, ethanol treatment, antibiotic treatment, gamma-radiation, and washing with deionized water on tailings characteristics and concentrations of IOB and SOB. Most probable number enumeration indicates that IOB and SOB were present at very low concentrations or below detection limits following treatment with all methods except rinsing and antibiotics treatment, where higher concentrations of IOB and SOB were present. The physical, chemical, and mineralogical characterization of the tailings indicated no changes in bulk mineralogy or bulk chemical composition as a result of treatment. However, an increase in oxidized sulfur species at the tailings surface, as determined by X ray photoelectron spectroscopy, was observed for the heating, autoclaving, and antibiotics treatments. Batch weathering experiments, used to evaluate the effect of treatment on element release rates, indicated that the final element release rates (after >30 d) were similar between treated and untreated control samples. On the basis of the results of this study, experiments over relatively long periods (>30 d) are to be recommended forthe establishment of microbial and abiotic weathering rates in mill tailings samples. For the determination of abiotic reaction rates, treatment by gamma-radiation is suggested to be the most appropriate method for sulfide-rich tailings. PMID- 15757339 TI - Complexation of heavy metals by phytochelatins: voltammetric study of the binding of Cd2+ and Zn2+ ions by the phytochelatin (gamma-Glu-Cys)3Gly assisted by multivariate curve resolution. AB - The complexation of Cd2+, Zn2+, and both together with the phytochelatin (gamma Glu-Cys)3Gly is studied by differential pulse polarography, and data are analyzed by multivariate curve resolution by alternating least squares (MCR-ALS). MCR-ALS yields the respective unitary voltammograms and concentration profiles of the resolved components, which contain information on the relative stabilities and stoichiometries of the formed complexes. The analysis of these results shows, for the Cd2+/(gamma-Glu-Cys)3Gly system, the presence of different kinds of bound Cd2+. For the Zn2+/ (gamma-Glu-Cys)3Gly system, the poor definition of the reduction signals of the complexes prevents a clear discrimination among differently bound Zn2+ ions. Atentative complexation/ electrochemical model is proposed for when both metal ions, Cd2+ and Zn2+, compete toward complexation, and some of the corresponding equilibrium constants are estimated. PMID- 15757340 TI - Regional assessment of the response of the acid-base status of lake watersheds in the Adirondack region of New York to changes in atmospheric deposition using PnET BGC. AB - Understanding the response of soil and surface waters to changes in atmospheric deposition is critical for guiding future legislation on air pollution. The Adirondack region of New York experiences among the most severe ecological impacts from acidic deposition. The region is characterized by considerable variability in atmospheric deposition, surficial and bedrock geology, hydrologic flow paths, and vegetation resulting in variability in effects of acidic deposition. In this study, an integrated biogeochemical model (PnET-BGC) was applied to 37 forest lake watersheds to assess the response of soil and surface waters of the Adirondacks to changes in atmospheric deposition at a regional scale. Model-simulated surface water chemistry was validated against data from two synoptic surveys conducted in 1984 and 2001. Results indicate that the model is able to capture the observed changes in surface water chemistry during this period. The model was further used to forecast the response of soil and surface waters to three future emission control scenarios. Results indicate that under the Clean Air Act, surface water SO4(2-) concentrations will continue to decrease at a median rate of -0.38 microeq/L-yr, and surface water ANC is predicted to increase at a median rate of 0.11 microeq/L-yr. More aggressive emission reductions will accelerate the rate of recovery. Under an aggressive control scenario, which represents an additional 75% reduction in SO2 emissions beyond the implementation of the Clean Air Act, surface water SO4(2-) concentrations are predicted to decrease at a median rate of -0.88 microeq/L-yr, and surface water ANC is predicted to increase at a median rate of 0.43 microeq/L-yr. Model predictions of several biologically relevant chemical indicators are also reported. PMID- 15757341 TI - Effect of nitrogen-containing compounds on polychlorinated dibenzo-p dioxin/dibenzofuran formation through de novo synthesis. AB - An experimental study was conducted to clarify the suppression effect of nitrogen containing compounds, that is, ammonia and urea, on the formation of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) through the de novo synthesis reaction. In the experiment, graphite and copper chloride contained in a mixture were used as sources of carbon and chlorine, respectively. The granulated sample mixture was charged as a packed-bed in the glass tube and heated at 300 degrees C in the flow of Ar-O2 gas mixture. In some cases, urea was added as aqueous solution to the sample, while ammonia was added to the gas flowed through the sample bed. The amount of PCDD/Fs formed decreases significantly by the addition of both ammonia and urea. Particularly, the addition of urea reduces the amount of PCDD/Fs discharged in the outlet gas by approximately 90%. The oxidation rate of carbon in the early stage of the experiment, that is, the heating period, is promoted bythe addition of nitrogen containing compounds. However, soon afterthe temperature reaches 300 degrees C, the formation rate becomes lower than that of the case without the addition of nitrogen-containing compounds. On the other hand, organic compounds containing amino (-NH2) or cyanide (-CN) groups and those containing nitrogen within the carbon ring frame were detected in the outlet gas in the case of urea addition. Typically observed aromatic compounds are chlorobenzonitriles, chlorobenzeneamines, and chloropyridines. This suggests a possibility that hydrogen and/or chlorine combined with PCDD/Fs are also substituted by such nitrogen-containing groups, and this decreases the formation rate of their frame of carbon rings. This phenomenon was also consistent with the fact that a significant reduction was observed in the amount of PCDD/Fs released to the outlet gas when urea was added. PMID- 15757342 TI - Glass transitions in peat: their relevance and the impact of water. AB - This contribution aims to expand the macromolecular view of fractionated natural organic matter (NOM)to organic matter in whole soils. It focuses on glass transition behavior of whole soil organic matter (SOM) and its interrelation with water through use of differential scanning calorimetry (DSC) and thermomechanical analysis (TMA). Three processes of structural relaxation related to macromolecular mobility were distinguished. Process I occurs in thermally pretreated and very low water-content samples and corresponds to classic glass transition behavior. Process II occurs in water-containing samples, where water is believed to act as an antiplasticizing agent in the peat at water contents below 12%, causing decreased macromolecular mobility and increased glass transition temperature. We suggestthe formation of hydrogen bond-based cross links being responsible for this antiplasticizing effect. Process III represents a slow swelling process induced by water uptake with a time constant of swelling in the order of days, with water acting as a plasticizing agent. Results from this work are of particular importance for environmental systems as changes in environmental conditions (e.g., water content, temperature) may induce slow structural relaxation processes in NOM over periods of time ranging from days to weeks. These influences on NOM macromolecular mobility lead to continuous changes in physicochemical properties that may greatly influence sorbate-sorbent interactions in surface and subsurface environments. PMID- 15757343 TI - Aggregation kinetics of kaolinite-fulvic acid colloids as affected by the sorption of Cu and Pb. AB - Aggregation kinetics of kaolinite-fulvic acid colloids, as influenced by two strongly sorbing trace metal cations, Cu(II) and Pb(II), was investigated by time resolved dynamic laser light scattering experiments. The effects of Cu and Pb on the aggregation rate and electrophoretic mobilitywere compared with that of Ca, another major divalent metal cation which is less strongly adsorbed. Kaolinite fulvic acid suspensions (in 0.01 M NaNO3 at pH 4 and pH 6) were spiked with solutions containing Cu, Pb, or Ca to give total divalent cation concentrations between 10(-5) and 8 x 10(-3) M. The concentration of kaolinite was varied between 25 and 200 mg L(-1), while the concentration of fulvic acid ranged from 0.15 to 1.2 mg L(-1). The mass ratio of kaolinite to fulvic acid was kept constant at 500:3 in all experiments. Relative aggregation rates, expressed as attachment efficiency alpha, were determined from linear increases in average hydrodynamic radius with time during the first 5-8 min of the aggregation experiments, always starting with a well-dispersed suspension at time zero. The corresponding slope for fast aggregation (alpha = 1) was measured for pure kaolinite suspended in 0.01 M NaNO3 at pH 4. Addition of fulvic acid to the suspensions completely inhibited kaolinite aggregation at pH 4 and pH 6. Additions of Cu, Pb, and Ca resulted in strongly increased aggregation rates of the kaolinite-fulvic acid particles. The potential of the three cations to enhance aggregation of the kaolinite-fulvic acid colloids increased in the order Ca < Cu < or = Pb. At pH 4, the relationship between particle electrophoretic mobility and aggregation rate was the same for all three divalent metal cations. In the presence of Ca, an increase in pH from 4 to 6 resulted in decreased aggregation rates. However, in the presence of Cu or Pb, the opposite trend was observed and the relationship between electrophoretic mobility and aggregation rate was different than at pH 4. The effects of Cu, Pb, and Ca on the aggregation rates of kaolinite-fulvic acid colloids are explained by the different sorption behavior of the three divalent metal cations. PMID- 15757344 TI - Tropospheric reaction of OH with selected linear ketones: kinetic studies between 228 and 405 K. AB - The absolute rate coefficients for the tropospheric reactions of hydroxyl radical (OH) with a series of linear aliphatic ketones (2-butanone (k1), 2-pentanone (k2), 2-hexanone (k3), and 2-heptanone (k4)) were measured as a function of temperature (228-405 K) and pressure (45-600 Torr of He) by the pulsed laser photolysis/laser induced fluorescence technique. These studies are essential to model the atmospheric chemistry of these ketones and their impact in the air quality. No pressure dependence of the rate coefficients was observed in the range studied. Thus, k1(298 K) (x10(-12) cm3 molecule(-1) s(-1)) were averaged over the pressure range studied yielding the following: (1.04+/-0.74), (3.14+/ 0.40), (6.37+/-1.40), and (8.22+/-1.10), for 2-butanone (k1), 2-pentanone (k2), 2 hexanone (k3), and 2-heptanone (k4), respectively. k1 exhibits a slightly positive temperature dependence over the temperature range studied. A conventional Arrhenius expression describes the observed behavior. In contrast, the temperature dependence of k2-k4 shows a distinct deviation from the Arrhenius behavior. The best fit to our data was found to be described by the three parameter expression: k(T) = A + B exp(-C/T) in cm3 molecule(-1) s(-1). This work constitutes the first determination of the temperature dependence of k2-k4. Our results are compared with previous studies, when possible, and are discussed in terms of the H-abstraction by OH radicals. The atmospheric implications of these reactions are also discussed. PMID- 15757345 TI - Oxidation of ferrocyanide by birnessite. AB - The Fe-CN complexes ferrocyanide, [FeII(CN)6]4-, and ferricyanide, [FeIII(CN)6]3 , which are contaminants in soil and groundwater, form a redox couple, [FeII(CN)6]4- <==> [FeIII(CN)6]3- + e-, E(H) = 356 mV. We studied the oxidation of [FeII(CN)6]4- by birnessite, delta-MnIVO2, in batch experiments as influenced by [FeII(CN)6]4- concentration, pH, and reaction time. Additionally, stopped-flow experiments were carried out at five temperatures (10-30 degrees C) and four pH values (pH 4.1-5.3). In the batch experiments, [FeII(CN)6]4- was completely oxidized to [FeIII(CN)6]3-, and oxidation did neither depend on time for t > 2 min, nor on concentration (0.12-0.47 mM), nor on pH (pH 3.3-9.9). Lasting adsorption of Fe-CN complexes on the birnessite surface or precipitation of manganese ferricyanide were not detected. Manganous ions resulting from the reductive dissolution of birnessite did not precipitate as manganese oxide because an identical decrease of Mn solution concentrations was observed under air and under a N2 atmosphere. Two processes were detected by the stopped-flow experiments. The first rapid one with an activation energy of approximately 60 kJ mol(-1) was attributed to short-term adsorption and simultaneous oxidation of [FeII(CN)6]4- on the birnessite surface. The second slower process with an activation energy of approximately 20 kJ mol(-1) was attributed most probably to diffusion of the reaction product Mn2+ into the interior of the birnessite, which creates fresh reaction sites at the outer surface. PMID- 15757346 TI - Emissions of metals associated with motor vehicle roadways. AB - Emissions of metals and other particle-phase species from on-road motor vehicles were measured in two tunnels in Milwaukee, WI during the summer of 2000 and winter of 2001. Emission factors were calculated from measurements of fine (PM2.5) and coarse (PM10) particulate matter at tunnel entrances and exits, and effects of fleet composition and season were investigated. Cascade impactors (MOUDI) were used to obtain size-resolved metal emission rates. Metals were quantified with inductively-coupled plasma mass spectrometry (ICP-MS) and X-ray fluorescence (XRF). PM10 emission rates ranged from 38.7 to 201 mg km(-1) and were composed mainly of organic carbon (OC, 30%), inorganic ions (sulfate, chloride, nitrate, ammonium, 20%), metals (19%), and elemental carbon (EC, 9.3%). PM10 metal emissions were dominated by crustal elements Si, Fe, Ca, Na, Mg, Al, and K, and elements associated with tailpipe emissions and brake and tire wear, including Cu, Zn, Sb, Ba, Pb, and S. Metals emitted in PM2.5 were lower (11.6% of mass). Resuspension of roadway dust was dependent on weather and road surface conditions, and increased emissions were related to higher traffic volumes and fractions of heavy trucks. Emission of noble metals from catalytic converters appeared to be impacted by the presence of older vehicles. Elements related to brake wear were impacted by enriched road dust resuspension, but correlations between these elements in PM2.5 indicate that direct brake wear emissions are also important. A submicrometer particle mode was observed in the emissions of Pb, Ca, Fe, and Cu. PMID- 15757347 TI - Exposure-pathway models explain causality in whole sediment toxicity tests. AB - Measurements of lethal effect concentrations (LC50) and bioaccumulation following water-only and whole-sediment exposures of the amphipod, Melita plumulosa, and the bivalve, Tellina deltoidalis, to copper, were combined with bioenergetic based kinetic models of exposure pathways to explain causality in whole-sediment toxicity tests. For both organisms, lethal body concentrations (LBCs) were greater for water-only exposures than for sediment exposures and indicated that the rate of copper accumulation and/or the mode of toxicity of copper assimilated were different for dissolved and particulate phases. The net assimilation of copper, expressed as a lethal exposure concentration (LEC) that was independent of the postexposure copper efflux, was shown to better explain the observed toxicity. The LEC of copper was the same for both water-only and whole-sediment toxicity tests. It is predicted that, for each species, a large range of effect concentrations may be measured for sediments having the same total copper concentration. These are conditional effect concentrations, as their value will be determined by total copper concentrations, partitioning (Kd) relationships (sediment properties), organism physiology (uptake rates from waters, assimilation efficiencies from solids), and organism feeding behavior (feeding selectivity). The importance of these factors to the development of sediment quality guidelines for metals based on species sensitivity distributions is discussed. PMID- 15757348 TI - New model calculations of pH-depending tributyltin adsorption onto montmorillonite surface and montmorillonite-rich sediment. AB - Interactions of the pollutant tributyltin (TBT) with mineral surfaces affects its distribution and transport in aqueous systems. In the present work, model calculations are reported that quantify TBT adsorption onto pure-phase montmorillonite (sample SWy) under various pH and salinity conditions that are important from an environmental perspective. The pH level in the system is of substantial interest because it affects the speciation of TBT in solution as well as the surface properties of the solid phase, which are both important for adsorption reactions. The model is based on the generalized diffused layer model that includes >X- sites in order to account for the cation exchange effects of TBT attraction. The presence of >AIOH and >SiOH sites atthe mineral surface was not considered separately during calculation. Instead, nonselective sites (>SOH) versus selective sites (>SsOH) were distinguished with respect to the sorptive sites on montmorillonite. The latter are characterized by a high affinity of TBT bonding. Both sorptive sites exhibit the same protolysis constants but different TBT binding constants [logK(TBT/2) = -1.18 for (>SOH), logK(TBT/1) = 3.98 for (>SsOH)]. LogK(X/TBT) for the cation exchange reaction was determined as between 3.05 and 4.14. The results indicate that the inclusion of selective sites during calculations is essential for quantifying pH-dependent TBT adsorption successfully. The parameters determined for the TBT adsorption onto pure-phase montmorillonite were subsequently used to calculate pH-dependent TBT adsorption onto a natural montmorillonite-rich sediment. PMID- 15757349 TI - Numerical model for biological fluidized-bed reactor treatment of perchlorate contaminated groundwater. AB - Biological fluidized-bed reactor (BFBR) treatment with 1.3 mm granular activated carbon as support medium is being used for removal of 2.6 mg/L perchlorate from contaminated groundwater in California. The California drinking-water action level of 4 microg/L for perchlorate requires 99.9% perchlorate removal. Sufficient ethanol, the electron donor, is added to remove oxygen and nitrate as well as perchlorate, as all three serve as electron acceptors, but with biological preference for oxygen and nitrate. A numerical BFBR model based upon basic physical, chemical, and biological processes including reaction stoichiometry, biofilm kinetics, and sequential electron acceptor usage was developed and evaluated with the full-scale treatment results. A key fitting parameter was bacterial detachment rate, which impacts reaction stoichiometry. For best model fit this was found to vary between 0.062 and 0.31 d(-1), with an average of 0.22 d(-1). The model indicates that GAC particle size, reactor diameter, and perchlorate concentration affect BFBR performance. While empty-bed detention time might be decreased somewhat below 10 min by an increase in either GAC particle size or reactor diameter, the current design provides a good factor of safety in operation. With a 10 min detention time, the effluent goal of 4 microg/L should be achievable even with influent perchlorate concentration as high as 10 mg/L. PMID- 15757350 TI - Risk assessment of short-chain chlorinated paraffins in Japan based on the first market basket study and species sensitivity distributions. AB - The chemical risks of short-chain chlorinated paraffins (SCCPs) to human health and ecological species in Japan were assessed in this study. SCCPs are used as extreme pressure additives in metal-working fluids and flame retardant agents in plastic materials. The first market basket study with a high resolution gas chromatography and mass spectrometry with electron capture negative ionization is reported. Total daily food intakes for Japanese residents of different ages were estimated using Latin Hypercube simulations, and the highest 95th percentile intake is 6.8 x 10(2) ng/kg/day for a 1-year-old child. Based on the reported no observed-adverse-effect-level (NOAEL) and the estimated exposure mainly from food, it was concluded thatSCCPs are not posing risks to humans in Japan. Ecological risks to aquatic and sediment organisms were estimated using species sensitivity distributions. The 95th percentiles of the distributions obtained by fitting several environmental SCCP concentration data of river water and sediment were 41.8 ng/L and 558 ng/g wet weight as the possible highest concentrations, respectively. No-observed-effect-concentrations (NOEC) were determined by calculating the fifth percentiles of a species sensitivity distribution, which were 2.2 microg/L for aquatic organisms and 1.7 to 13.5 mg/ kg wet weight for sediment organisms. While it is likely that there is no imminent environmental chemical risk for aquatic organisms at a regional level in Japan, this study concluded that a more detailed risk assessment is necessary for sediment organisms. PMID- 15757351 TI - Sediment depth attenuation of biogenic phosphorus compounds measured by 31P NMR. AB - Being a major cause of eutrophication and subsequent loss of water quality, the turnover of phosphorus (P) in lake sediments is in need of deeper understanding. A major part of the flux of P to eutrophic lake sediments is organically bound or of biogenic origin. This P is incorporated in a poorly described mixture of autochthonous and allochthonous sediment and forms the primary storage of P available for recycling to the water column, thus regulating lake trophic status. To identify and quantify biogenic sediment P and assess its lability, we analyzed sediment cores from Lake Erken, Sweden, using traditional P fractionation, and in parallel, NaOH extracts were analyzed using 31P NMR. The surface sediments contain orthophosphates (ortho-P) and pyrophosphates (pyro-P), as well as phosphate mono- and diesters. The first group of compounds to disappear with increased sediment depth is pyrophosphate, followed by a steady decline of the different ester compounds. Estimated half-life times of these compound groups are about 10 yr for pyrophosphate and 2 decades for mono- and diesters. Probably, these compounds will be mineralized to ortho-P and is thus potentially available for recycling to the water column, supporting further growth of phytoplankton. In conclusion, 31P NMR is a useful tool to asses the bioavailability of certain P compound groups, and the combination with traditional fractionation techniques makes quantification possible. PMID- 15757352 TI - Feasibility of reflectance spectroscopy for the assessment of soil mercury contamination. AB - Conventional methods for investigating soil Hg contamination based on raster sampling and chemical analysis are time-consuming and relatively expensive. The objective of this study was to develop a rapid method for investigating Hg concentration in suburban agricultural soils of the Nanjing region using reflectance spectra within the visible-near-infrared (VNIR) region. Several spectral pretreatments (absorbance, Kubelka-Munk transformations and their derivatives) were applied to the reflectance spectra to optimize the accuracy of prediction. The prediction of Hg concentration was achieved by univariate regression and principal component regression (PCR) approaches. The optimal model (R= 0.69, RMSEP = 0.15) for predicting Hg was achieved using the PCR method with the Kubelka-Munktransformation asthe spectral predictor. Comparison of three wavelength ranges (0.38-1.1, 1.0-2.5, and 0.38-2.5 microm) on the effect of prediction accuracy showed that the best results were acquired using the 1.0-2.5 microm spectral region. Correlation analysis revealed that Hg concentration was negatively correlated with soil reflectance while positively correlated with the absorption depths of goethite at 0.496 microm and clay minerals at 2.21 microm, suggesting that Hg-sorption by clay-size mineral assemblages in soils was the mechanism by which to predict spectrally featureless Hg. These results indicate that it is feasible to predict Hg levels in agricultural soils using the rapid and cost-effective reflectance spectroscopy. Future study with operational remote sensing techniques and field measurements is strongly recommended. PMID- 15757353 TI - Dissolved organic nitrogen measurement using dialysis pretreatment. AB - Dissolved organic nitrogen (DON) is important for ecological and engineering researches. Quantification of low DON concentrations in waters with elevated dissolved inorganic nitrogen (DIN) using existing methods is inaccurate. In this study, a dialysis-based pretreatment technique was optimized and adopted to reduce the interference from DIN to the quantification of DON in natural water. A cellulose ester dialysis tube (nominal molecular weight cutoff = 100 Da) was used in batch and continuous-flow dialysis steps with model compounds, natural organic matter isolates, and bulk waters to develop a dialysis pretreatment approach that selectively reduces DIN from solutions containing DON. By reducing DIN concentrations, propagation of analytical variance in total dissolved nitrogen (TDN) and DIN species concentrations allows more accurate determination of DON (DON = TDN - NO3 - NO2- - NH3/NH4+). Dialysis for 24 h against continuously flowing distilled water reduced DIN species by 70%. With dialysis pretreatment, DON recoveries of more than 95% were obtained for surface water and finished drinking water, but wastewater experienced a slight loss (approximately 10%) of DON possibly due to the adsorption of organics onto the dialysis membrane, permeation of low molecular weight fractions, or biodegradation. Dialysis experiments using surface water spiked with different DIN/TDN ratios concluded that dialysis pretreatment leads to more accurate DON determination than no dialysis when DIN/TDN ratios exceed 0.6 mg of N/mg of N. PMID- 15757354 TI - Adsorption onto fluidized powdered activated carbon flocs-pACF. AB - This work presents a new adsorption technique where the adsorbent (powdered activated carbon-PAC) is in the form of suspended flocs formed with water-soluble polymer flocculants. Thus, the adsorption of a typical dye, methylene blue (MB), was studied onto polyacrylamide flocs of PAC (PACF) in a fluidized bed reactor. The technique is based on the fact that the adsorption capacity of PAC does not decrease after flocculation because the adsorbed polymer occupies only a few surface sites, in the form of trains, loops, and tails. Moreover, the adsorption was found to proceed through a rapid mass transfer of MB to the adsorbing PAC flocs, in the same extent as onto PAC. Because of the rapid settling characteristics of the aggregates formed, the two phase separations, loaded PAC and solution, become easier. Thus, the technique offers the advantages of conducting simultaneously both adsorption and solid/liquid separation all in one single stage. Results obtained showed that high MB removal values can be attained in a fluidized bed reactor (>90%) and that PACF presents a much higher adsorption capacity (breakthrough points) than granulated activated carbon (GAC) in the same adsorbing bed. It is believed that this technique highly broadens the potential of the use of powdered activated carbon or other similar ultrafine adsorbents. PMID- 15757355 TI - Hydraulic shear stress calculation in a sequencing Batch biofilm reactor with granular biomass. AB - This paper reports the results of an experimental study specifically aimed at developing a simple methodology for calculating hydrodynamic shear forces in a sequencing batch biofilm reactor (SBBR) system with granular biomass. Using such a methodology, the hydrodynamic shear forces are simply calculated by measuring bed porosity and pressure losses. In addition, by applying this methodology an explanation for the biomass evolution from biofilm to granules under aerobic conditions has been provided and the following mechanism has been proposed: (i) formation of a thin biofilm that fully covers the carrier; (ii) increase of biofilm thickness; (iii) break-up of the attached biofilm with release of biofilm particles; (iv) rearrangement of biofilm particles in smooth granules. The hydrodynamic shear forces trend during the start-up period provides an explanatory key for the generation process of granular biomass. In fact, during the first two steps, the SBBR is characterized by rather weak shear forces values (lower than 1 dyn/cm2). Under these weak shear forces, the biofilm grows by increasing its thickness through a porous structure and weak adhesion strengths. Such a continuous increase of biofilm thickness produces an increase of the shear forces with negative effect on biomass stability, causing the detachment of biofilm particles. In turn, such detachment causes a further sharp increase of shear forces (more than 10 times) that promotes the rearrangement of the detached biofilm particles in smooth granules. A correlation between biomass density and hydrodynamic shear forces was observed. In particular, the biomass density linearly increases with the increase of shear stress. PMID- 15757356 TI - Development of a biosorbent for arsenite: structural modeling based on X-ray spectroscopy. AB - This work describes a biological route for direct sorption of aqueous As(III) species, which are the most toxic and mobile arsenic species found in soils. Based upon the biochemical mechanisms that explain arsenic toxicity, we propose that a waste biomass with a high fibrous protein content obtained from chicken feathers can be used for selective As(III) adsorption. Prior to adsorption, the disulfide bridges present in the biomass are reduced by thioglycolate. Our investigations demonstrated that As(III) is specifically adsorbed on the biomass and, contrary to the behavior observed with inorganic sorbents, the lower is the pH the more effective is the removal. Arsenic uptake reaches values of up to 270 micromol As(III)/g of biomass. Analyses by synchrotron light techniques, such as XANES, demonstrated that arsenic is adsorbed in its trivalent state, an advantage over conventional techniques for As uptake, which usually require a previous oxidation stage. EXAFS analyses showed that each As atom is directly bound to three S atoms with an estimated distance of 2.26 A. The uptake mechanism is explained in terms of the structural similarities between the As(III)-biomass complex structure and that of arsenite ions and Ars-Operon system encoded proteins and phytochelatins. The biological route presented here offers the perspective of a direct removal of arsenic in its reduced form. PMID- 15757357 TI - Sorption and desorption of perchlorate and U(VI) by strong-base anion-exchange resins. AB - This study investigated the sorption affinity and capacity of six strong-base anion-exchange (SBA) resins for both uranium [U(VI)] and perchlorate (ClO4-) in simulated groundwater containing varying concentrations of sulfate (SO4(2-)). Additionally, desorption of U(VI) from spent resins was studied to separate U(VI) from resins with sorbed ClO4- for waste segregation and minimization. Results indicate that all SBA resins investigated in this study strongly sorb U(VI). The gel-type polyacrylic resin (Purolite A850) showed the highest sorption affinity and capacityfor U(VI) butwasthe least effective in sorbing ClO4-. The presence of SO4(2-) had little impact on the sorption of U(VI) but significantly affected the sorption of ClO4-, particularly on monofunctional SBA resins. A dilute acid wash was found to be effective in desorbing U(VI) but ineffective in desorbing ClO4- from bifunctional resins (Purolite A530E and WBR109). A single wash removed approximately 75% of sorbed U(VI) but only approximately 0.1% of sorbed ClO4- from the bifunctional resins. On the other hand, only 21.4% of sorbed U(VI) but approximately 34% of sorbed ClO4- was desorbed from the Purolite A850 resin. This study concludes that bifunctional resins could be used effectively to treatwater contaminated with ClO4- and traces of U(VI), and dilute acid washes could minimize hazardous wastes by separating sorbed U(VI) from ClO4- prior to the regeneration of the spent resin loaded with ClO4-. PMID- 15757358 TI - Comment on "Atmospheric mercury accumulation rates between 5900 and 800 calibrated years BP in the high arctic of canada recorded by peat hummocks". PMID- 15757359 TI - Creativity: self-perceptions over time. AB - This study explores creativity and motivation in the second half of life. While many quantitative studies on creativity show decline with advancing age, there is some evidence that creativity may change qualitatively over time. The purpose of this study is to elucidate self-perceived changes in creativity over the life span. The participants included visual artists, both working and retired. A total of 21 adults, ages 53 to 75, participated in the study. Ten of the artists were interviewed in depth regarding the perception of their own creativity over time. Another 11 artists participated in two separate focus groups. All of the participants represented their perceived creativity using an adaptation of the Creative Life-Line Curve (CLLC; Romaniuk, Romaniuk, Sprecher, & Cones, 1983). Findings indicated that participants do not perceive a decline in creativity with age. In-depth analysis of the interviews included qualitative coding to establish a number of themes that permeated the self-perceptions regarding changes in creativity over time. These themes were found to exemplify many of the concepts inherent in the life-span developmental perspective. PMID- 15757360 TI - Determinant factors contributing to variations in memory performance in centenarians. AB - The role of environmental markers in memory performance among centenarians was examined in a study comparing non-cognitive factors to various aspects of memory performance. The centenarian group participating in the study consisted of 100 individuals (82 females and 18 males). Selected as non-cognitive factors were Quality of Life, Life-Habits, Health, Personality, and Autonomy. These factors were later compared to memory performance in short-term memory, semantic memory, and episodic memory. Using a structural equation model, results showed that performance in short-term memory was best predicted by life-habits and personality, while performance in semantic and episodic memory were best predicted by autonomy and life-habits. Implementation of the result from a life span perspective includes a closer monitoring of environmental markers in the future. In effect, this could preserve memory function and optimum health through old age, thus making interventions easier to realize. PMID- 15757361 TI - Age differences in perceptions of control in social relationships. AB - To date, few studies have examined perceptions of control in interpersonal relationships, and whether such perceptions reflect individual characteristics (e.g., age) and features of relationships (e.g., quality). One hundred and eighty seven individuals aged 13 to 99 (M = 45.62, SD = 25.69) provided information about their close and problematic relationships and their perceptions of control over those relationships. Multilevel modeling revealed that both relationship features and individual characteristics were associated with perceptions of control in relationships. Specifically, individuals felt more control in their marital relationships, and in relationships characterized by higher quality and fewer worries. Age differences in individuals' perceptions of control in relationships appear to reflect age-related shifts in perceptions of control in the parent-child tie. PMID- 15757362 TI - Spiritual influences on ability to engage in self-care activities among older African Americans. AB - The influence of spiritual factors on the ability of African-American elders to carry out instrumental activities of daily living (IADL) independent of age, gender, education, and self-rated health is explored using the religion-health explanatory model in a cross-sectional sample of 96 African-American community dwelling adults 62 to 93 years of age. The Reed spiritual perspective (SPS) and self-transcendence (STS) scales are used to study spiritual factors (Reed, 1991). The typical respondent was 75 years of age, female, widowed with 10.4 years of education. Self-rated health and age are strongly related to IADL in models that include the other variables (R2 = 0.41, p < 0.01). Those who are younger and those who self-report better health have higher IADL scores than those without these characteristics. Spiritual factors are significantly related to IADL in a model that includes an interaction of STS with education (R2= 0.50). Among the least educated, STS is associated with higher scores of IADL. Addition of the interaction to the model resulted in a significant positive association of both STS and education with IADL. SPS was unrelated to IADL. Caregivers concerned with functional ability may want to consider interventions based on increasing a sense of self-transcendence, and to consider educational level as a potential moderator of this relationship. Criticism of the religion-health literature has suggested that putative health effects of religion may be exaggerated, because of failure to take confounding variables into account (Sloan, Bagiella, & Powell, 1999). However, this study and a recent survey by Musick, House, and Williams (2004) are evidence that it is just as likely that health benefits of religion would be hidden by confounders as that they would be exaggerated by them. PMID- 15757363 TI - Gray wolves as climate change buffers in Yellowstone. AB - Understanding the mechanisms by which climate and predation patterns by top predators co-vary to affect community structure accrues added importance as humans exert growing influence over both climate and regional predator assemblages. In Yellowstone National Park, winter conditions and reintroduced gray wolves (Canis lupus) together determine the availability of winter carrion on which numerous scavenger species depend for survival and reproduction. As climate changes in Yellowstone, therefore, scavenger species may experience a dramatic reshuffling of food resources. As such, we analyzed 55 y of weather data from Yellowstone in order to determine trends in winter conditions. We found that winters are getting shorter, as measured by the number of days with snow on the ground, due to decreased snowfall and increased number of days with temperatures above freezing. To investigate synergistic effects of human and climatic alterations of species interactions, we used an empirically derived model to show that in the absence of wolves, early snow thaw leads to a substantial reduction in late-winter carrion, causing potential food bottlenecks for scavengers. In addition, by narrowing the window of time over which carrion is available and thereby creating a resource pulse, climate change likely favors scavengers that can quickly track food sources over great distances. Wolves, however, largely mitigate late-winter reduction in carrion due to earlier snow thaws. By buffering the effects of climate change on carrion availability, wolves allow scavengers to adapt to a changing environment over a longer time scale more commensurate with natural processes. This study illustrates the importance of restoring and maintaining intact food chains in the face of large-scale environmental perturbations such as climate change. PMID- 15757364 TI - Functional evolution of a cis-regulatory module. AB - Lack of knowledge about how regulatory regions evolve in relation to their structure-function may limit the utility of comparative sequence analysis in deciphering cis-regulatory sequences. To address this we applied reverse genetics to carry out a functional genetic complementation analysis of a eukaryotic cis regulatory module-the even-skipped stripe 2 enhancer-from four Drosophila species. The evolution of this enhancer is non-clock-like, with important functional differences between closely related species and functional convergence between distantly related species. Functional divergence is attributable to differences in activation levels rather than spatiotemporal control of gene expression. Our findings have implications for understanding enhancer structure function, mechanisms of speciation and computational identification of regulatory modules. PMID- 15757366 TI - Porphyrin-fullerene C60 dyads with high ability to form photoinduced charge separated state as novel sensitizers for photodynamic therapy. AB - The photodynamic activities of a porphyrin-C60 dyad (P-C60) and its metal complex with Zn(II) (ZnP-C60) were compared with 5-(4-acetamidophenyl)-10,15,20-tris(4 methoxyphenyl)porphyrin (P), both in homogeneous medium-bearing photooxidizable substrates and in vitro on the Hep-2-human-larynx-carcinoma cell line. This study represents the first evaluation of dyads, with a high capacity to form a photoinduced charge-separated state, to act as agents to inactivate cells by photodynamic therapy (PDT). Absorption and fluorescence spectroscopic studies were performed in toluene and N,N-dimethylformamide (DMF). The emission of the porphyrin moiety in the dyads is strongly quenched by the attached fullerene C60 moiety. The singlet molecular oxygen, O2(1delta(g)), productions (phi(delta)) were determined using 9,10-dimethylanthracene (DMA). The values of phi(delta) were strongly dependent on the solvent's polarity. Comparable phi(delta) values were found for dyads and P in toluene, while O2(1delta(g)) production was significantly diminished for the dyads in DMF. In more polar solvent, the stabilization of charge-transfer state takes place, decreasing the efficiency of porphyrin triplet-state formation. Also, both dyads photosensitize the decomposition of L-tryptophan in DMF. In biological medium, no dark cytotoxicity was observed using sensitizer concentrations < or = 1 microM and 24 h of incubation. The uptake of sensitizers into Hep-2 was studied using 1 microM of sensitizer and different times of incubation. Under these conditions, a value of approximately 1.5 nmol/10(6) cells was found between 4 and 24 h of incubation. The cell survival after irradiation of the cells with visible light was dependent upon light-exposure level. A higher photocytotoxic effect was observed for P-C60, which inactivates 80% of cells after 15 min of irradiation. Moreover, both dyads keep a high photoactivity even under argon atmosphere. Thus, depending on the microenvironment where the sensitizer is localized, these compounds could produce biological photodamage through either an O2(1delta(g))-mediated photoreaction process or a free-radicals mechanism under low oxygen concentration. These results show that molecular dyads, which can form a photoinduced charge-separated state, are a promising model for phototherapeutic agents, with potential applications in cell inactivation by PDT. PMID- 15757365 TI - Adult murine skeletal muscle contains cells that can differentiate into beating cardiomyocytes in vitro. AB - It has long been held as scientific fact that soon after birth, cardiomyocytes cease dividing, thus explaining the limited restoration of cardiac function after a heart attack. Recent demonstrations of cardiac myocyte differentiation observed in vitro or after in vivo transplantation of adult stem cells from blood, fat, skeletal muscle, or heart have challenged this view. Analysis of these studies has been complicated by the large disparity in the magnitude of effects seen by different groups and obscured by the recently appreciated process of in vivo stem cell fusion. We now show a novel population of nonsatellite cells in adult murine skeletal muscle that progress under standard primary cell-culture conditions to autonomously beating cardiomyocytes. Their differentiation into beating cardiomyocytes is characterized here by video microscopy, confocal-detected calcium transients, electron microscopy, immunofluorescent cardiac-specific markers, and single-cell patch recordings of cardiac action potentials. Within 2 d after tail-vein injection of these marked cells into a mouse model of acute infarction, the marked cells are visible in the heart. By 6 d they begin to differentiate without fusing to recipient cardiac cells. Three months later, the tagged cells are visible as striated heart muscle restricted to the region of the cardiac infarct. PMID- 15757367 TI - Rejecting the label: a social constructionist analysis. AB - Research has shown that people often do not claim labels associated with mental retardation or learning difficulties. We discussed the interpretation that this rejection is an example of a denial process, the purpose of which is to protect self-esteem. Alternative explanations for this lack of identification were offered, based on an understanding of the socially constructed nature of diagnostic labels and on the distinction between diagnostic labels and social categories. Some of the problems in using the label as a descriptive or explanatory resource are illustrated using quotes from a study in which people who have been labeled discussed the label. PMID- 15757368 TI - Parents' attributions of their child's jigsaw-puzzle performance: comparing two genetic syndromes. AB - Parents' attributions of the jigsaw-puzzle performance of their child with Prader Willi syndrome (n = 20) or Williams syndrome (n = 21) were examined. Parents in both groups placed more importance on internal versus external attributions. Parents of children with Prader-Willi syndrome exhibited a hedonic bias by attributing their child's success to internal factors (particularly when the child was good at puzzles), downplayed the role of chance when their child had higher puzzle skills and higher cognitive functioning, and differentiated between internal and external attributions. In contrast, parents of children with Williams syndrome showed an inconsistent attributional pattern. These findings help identify how parents understand the intellectual performance of their child with disabilities and how interventions might be beneficial. PMID- 15757369 TI - Simple decency. PMID- 15757370 TI - "New" state information systems; same "old" mistakes. PMID- 15757371 TI - Fiscal year 2004 institution populations, movement, and expenditures by state with national comparisons to earlier years. PMID- 15757374 TI - Factors influencing direct-care paraprofessionals' decisions to initiate mental health referrals for adults with mental retardation. AB - Direct-care paraprofessionals' recognition of psychopathology of varying severity in persons with mental retardation was evaluated. Factors that may influence paraprofessionals' decisions to initiate referrals for mental health services on behalf of individuals with mental retardation were also evaluated. Results suggest that staff members recognized and differentiated psychopathology of varying levels of clinical severity. Results also suggest that paraprofessionals are more likely to initiate making a referral when professionals are perceived as being competent in treating individuals with mental retardation, and when providers' interventions are consistent with the referring agency's philosophy. PMID- 15757375 TI - Importance of experimental as well as empirical qualitative studies in special education. AB - In the past few decades qualitative research has increasingly appeared in special education journals. However, much of this work falls within the parameters of producing useful technical information that can be applied to the contexts where children and adults with disabilities learn, work, and live. Experimental qualitative studies that rely on postmodern or poststructural analyses, critical theory, and narrative research with subjective personal stories seem to be considered too radical, ideological, and theoretical to make it into many special education scholarly outlets. We argue that experimental qualitative designs have much to contribute to the fields of special education and disability studies and, hence, should reach those who receive or provide services to people with disabilities. PMID- 15757376 TI - Medicinally useful proteins--enhancing the probability of technical success in the clinic. AB - Proteins that are normally present in organisms function to maintain the overall homeostatic balance of the living system. Although an understanding of the biology of these proteins is essential to determining their utility in the treatment of disease, the administration of wild-type forms of proteins in a therapeutic manner (often systemically and at concentrations much higher than those found in the healthy organism) can have the effect of inappropriate activation of cell types and compartments that can lead to an undesirable response. Modification of the sequence of the parent protein to yield selected biological properties can generate derivative versions (muteins) having an optimal therapeutic profile. In addition to consideration of the biological properties, protein therapeutics are typically not active when delivered orally. This can limit the frequency with which the therapeutic agent can be administered to the patient, exemplifying the need to understand the pharmacokinetic and pharmacodynamic relationships of the protein. The biophysical properties of the mutein (e.g., solubility and stability) must also be considered in this context. An additional consideration with therapeutic proteins, engineered or not, is the potential immune response resulting from sequence variation and/or exogenous presentation to the immune system. The composite application of these principles to therapeutic protein discovery and development drives the ultimate success of any given biopharmaceutical agent. PMID- 15757377 TI - Ethical and regulatory aspects of embryonic stem cell research. AB - Ethical issues concerning human embryonic stem cell (hESC) research are reviewed three years after the controversy became a public and political issue in the US and two years after the last article on this topic, which also covered the religious aspects. There are still no consistent regulations worldwide on the regulation of research on hESCs, and debate is still ongoing in some countries. New developments require a reassessment of the situation. In the US, the debate is politically driven. Regulations still vary from one country to another and there are no universally uniform ethical and regulatory standards for hESC research. At present, the most liberal and favourable environments for hESC research are in the UK, Singapore, Sweden, India, Israel and China. In the US, there is no change in the policy of the current government, but developments in the industrial sector and academic institutions, which are not dependent on government funding, are encouraging for future progress. Considerable advances have taken place in hESC research during the past few years to justify a relaxation of restrictions, and suggestions are offered to improve the ethical and regulatory environments for hESC research. PMID- 15757378 TI - Natural killer cells: can they be useful as adoptive immunotherapy for cancer? AB - As part of the innate immune system, natural killer (NK) cells form the first line of defence against pathogens or transformed/cancerous host cells. Recent experimental and clinical data show the possibility of exploiting NK activity as a cell-based immunotherapy to treat cancer. This review discusses the recent knowledge on NK cell biology that has impacted on its development as a treatment for cancer. PMID- 15757379 TI - RNA transfer and its use in dendritic cell-based immunotherapy. AB - RNA is a key macromolecule for the mobilisation and interpretation of genetic information. Research has sought to exploit the inherent properties of RNA, such as the direct production of proteins in the cytoplasm without the need for nuclear translocation. This property makes the delivery of genes into postmitotic cells especially attractive. Recently, RNA transfer into postmitotic dendritic cells (DCs) has emerged as a potential new therapeutic agent in the area of immunotherapy. DCs are the most important regulators of the immune system. Thus, transfecting DCs with RNA allows the specific manipulation of immune responses and, thereby, the treatment of a variety of diseases, such as cancer. Preclinical studies have demonstrated that RNA-transduced DCs efficiently stimulate antigen specific T cell responses in vitro and in animal tumour models. In addition, the clinical data from Phase I and II trials of tumour patients indicate that RNA transduced DCs represent a promising approach for the development of future vaccination strategies. The use of RNA molecules as therapeutic agents is a relatively new approach in the treatment of diseases, such as cancer, but has received increasing attention during the past decade. Especially in the field of immunotherapy, the inherent properties of RNA molecules in combination with immunostimulating dendritic cells (DCs) are being investigated at present for their beneficial therapeutic effect. Immunotherapy is based on the stimulation of the patient's immune system to recognise and eliminate infected cells or tumour cells in an antigen-specific manner. Current approaches focus on the stimulation of CD8(+) cytotoxic T lymphocyte responses, as well as on the induction of CD4(+) T helper cell responses, in order to obtain optimal and sustained immune responses capable of eliminating altered cells. This review mainly focuses on the potential use of RNA-transduced DCs as a therapeutic strategy in the treatment of cancer, as current studies on the treatment of infectious diseases are just beginning. PMID- 15757380 TI - Gene therapy for the treatment of mitochondrial DNA disorders. AB - Despite recent epidemiological studies confirming that mitochondrial respiratory chain disorders due to mutations in either the mitochondrial or nuclear genome are amongst the most common inherited human diseases, realistic therapeutic strategies for these patients remain limited. The disappointing response to various vitamins, cofactors and electron acceptors that have been administered to patients in an attempt to bypass the underlying respiratory chain defect, coupled with the complexities of human mitochondrial genetics, means that novel and innovative means are required to offer realistic treatments. Several 'gene therapy' strategies have therefore been proposed to treat patients with pathogenic mitochondrial DNA mutations, and although these are not without their own inherent problems, several exciting approaches promise much in the near future. This review will provide a basic background to mitochondrial genetics and mitochondrial DNA disorders before introducing the various strategies being tested in vitro at present, in cell culture and animal models, and, in the example of therapeutic exercise, in patients themselves. PMID- 15757381 TI - Development of mammalian artificial chromosomes for the treatment of genetic diseases: Sandhoff and Krabbe diseases. AB - Mammalian artificial chromosomes (MACs) are being developed as alternatives to viral vectors for gene therapy applications, as they allow for the introduction of large payloads of genetic information in a non-integrating, autonomously replicating format. One class of MACs, the satellite DNA-based artificial chromosome expression vehicle (ACE), is uniquely suited for gene therapy applications, in that it can be generated denovo in cells, along with being easily purified and readily transferred into a variety of recipient cell lines and primary cells. To facilitate the rapid engineering of ACEs, the ACE System was developed, permitting the efficient and reproducible loading of pre-existing ACEs with DNA sequences and/or target gene(s). As a result, the ACE System and ACEs are unique and versatile platforms for ex vivo gene therapy strategies that circumvent and alleviate existing safety and delivery limitations surrounding conventional gene therapy vectors. This review will focus on the status of MAC technologies and, in particular, the application of the ACE System towards an ex vivo gene therapy treatment of lysosomal storage diseases, specifically Sandhoff (MIM #268800) and Krabbe (MIM #245200) diseases. PMID- 15757382 TI - Antisense approaches for inhibiting respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) continues as an emerging infectious disease not only among infants and children, but also for the immune-suppressed, hospitalised and the elderly. To date, ribavirin (Virazole, ICN Pharmaceuticals, Inc.) remains the only therapeutic agent approved for the treatment of RSV. However, its clinical benefits are small and occur only in a fraction of RSV-infected patients. The prophylactic administration of palivizumab (Synagis, MedImmune, Inc.) is problematic and costly and, therefore, only recommended for use in high risk infants. Clearly, the need for an effective and safe drug remains high. This review discusses several different antisense approaches and compares them with traditional strategies, such as RSV-targeting antibodies and antivirals, as well as developments in vaccine research. PMID- 15757383 TI - siRNA as a route to new cancer therapies. AB - The RNA interference (RNAi) gene-silencing mechanism is induced by double stranded RNA (dsRNA) and is highly sequence-specific. It is an extremely powerful tool for silencing gene expression in vitro, and might be used as therapy in human pathologies such as cancer, viral infections and genetic disorders. RNAi was initially discovered in plants, but it has become clear that it is also conserved in animal species. Triggering of RNAi by the introduction of small dsRNA (or small interfering RNA) into living cells as a tool to inhibit the expression of specific genes holds the promise to selectively extinguish the expression of disease-associated genes in humans. On the other hand, RNAi technology will serve to elucidate the functions and interactions of the thousands of human genes in high-throughput systems and help in target validation technology. PMID- 15757384 TI - Recent developments in the production of human therapeutic proteins in eukaryotic algae. AB - Antibody-based therapeutics have had great success over the last few years, and continue to be one of the fastest growing sectors of drug development. The efficacy and specificity of antibody-based drugs makes them ideal candidates for new drug development, but the specificity of these drugs comes from their complexity, and this complexity makes antibodies very expensive to produce. To address this problem, the authors have developed a system for the expression of recombinant proteins using the unicellular eukaryotic green algae, Chlamydomonas reinhardtii. As proof of concept, the authors have engineered microalgae to produce several forms of a human IgA antibody directed against herpes simplex virus. The expression of human monoclonal antibodies in C. reinhardtii offers an attractive alternative to traditional mammalian-based expression systems, as both the plastid and nuclear genomes are easily and quickly transformed, and the production of proteins in algae has an inherently low cost of capitalisation and production. PMID- 15757385 TI - TransMabs: cell-penetrating antibodies, the next generation. AB - Intracellular proteins are becoming attractive targets in diagnosis and for therapy such as in signal pathways, on enzymes, transcription factors and structural proteins. Antibodies have been used therapeutically for extracellular pathogens and for targeting cell-surface antigens. Antibodies normally do not pass easily through intact cellular or subcellular membranes in living cells. Methods to shuttle antibodies into living cells are either labour-intensive and/or compromise the structural and functional integrity of the cell or require the integration of genes for heavy and light chain production through gene therapy approaches. A new technology platform, 'SuperAntibody Technology', enables antibodies to be shuttled into living cells without harming them. Such cell-penetrating antibodies open new diagnostic and therapeutic windows. The term 'TransMabs' has been coined for these antibodies. Proof of principle has been achieved with a 17-amino acid peptide with membrane translocating properties, conjugated with anti-caspase-3 antibody. Such a TransMab inhibits significantly in vitro apoptosis-related events, such as caspase-3 activity, DNA fragmentation and spectrin cleavage. Anti-caspase-3 TransMab, therefore, could be utilised to inhibit apoptosis in a variety of diseases, such as Alzheimer's, Huntington's and Parkinson's. Unlike peptide inhibitors available at present, this TransMab is not expected to have invivo toxic side effects and can only target activated forms of the enzyme. This paper discusses the advantages and limitations of cell penetrating antibodies (TransMabs) compared with existing small molecule drug development approaches. PMID- 15757386 TI - Role of anti-C1q autoantibodies in the pathogenesis of lupus nephritis. AB - Anti-C1q autoantibodies can be found in the sera of patients with several autoimmune diseases, but also in healthy individuals. Although these anti-C1q autoantibodies were already identified several decades ago, they still puzzle both immunologists and nephrologists. The main reason for this puzzling effect are observations that seemed to indicate quite clearly that anti-C1q should be pathogenic to the kidney and the observation on the other hand that anti-C1q autoantibodies can be found in several disease conditions, as well as in healthy individuals, and are then unrelated to overt renal inflammation. This puzzle is the focus of the current review, which will provide an overview of the historical data, define the clinical interests and, importantly, will try to put several aspects in perspective based on recent observations in patients and in murine models. In addition, the paper will discuss therapeutic intervention possibilities regarding anti-C1q-mediated damage in systemic lupus erythematosus, as well as the therapeutic potential of anti-C1q antibodies in other conditions. PMID- 15757387 TI - From mice to humans - murine intelligence for human CD8+ T cell vaccine design. AB - There has been considerable progress in the design of vaccines capable of safely and effectively inducing CD8(+) T cells for prophylaxis and treatment of chronic infectious diseases and cancer. Much of what is known about CD8(+) T cell mediated immunity has come from pioneering work in mice; this broad overview discusses recent work in mouse systems where lessons may be drawn for human vaccine development. The areas highlighted include antivector immunity, immunodominance, dendritic cell biology and targeting, the role of Toll-like receptors and their exploitation by novel adjuvants, the role of CD4(+) T cell help, regulatory T cells and, finally, some comments on the different requirements of prophylactic versus therapeutic vaccines. PMID- 15757388 TI - Recent developments in Neisseria meningitidis group A conjugate vaccines. AB - Meningococcal disease, both endemic and epidemic, remains a major cause of meningitis in many countries. Protective immunity is mediated primarily by bacteriocidal antibodies against the capsular polysaccharides for serogroups other than B, and against non-capsular surface components for group B. This article focuses on the development of conjugate vaccines for serogroup A, with special emphasis on the needs of Africa. The first licensed (1999) meningococcal conjugate was against group C in the UK and was > 90% effective in infants, children and young adults. The problem now is to develop a highly immunogenic group A meningococcal conjugate vaccine for use in developing countries as an alternative to the presently licensed group AC polysaccharide vaccine. Immunogenicity studies on the group A polysaccharide show the polysaccharide itself to be uniquely immunogenic in young children compared with other polysaccharides, making comparative studies with a highly immunogenic conjugate of considerable importance. PMID- 15757389 TI - Innovative antibacterial drugs: nothing ventured, nothing gained. AB - The current paradox of the widespread clinical demand for novel anti-bacterial drugs and lack of commensurate industrial research activity is unprecedented in modern drug discovery. Although the root causes of the pharmaceutical industry's disengagement from antibacterial drug discovery have been discussed at length and attributed largely to a lack of productivity coupled with pharmacoeconomic and regulatory considerations, the challenges faced by privately held biotech companies in addressing this therapeutic need originate from the conflict between the capital intensive nature of early-stage drug discovery and the current venture capital funding paradigm. Continued innovation of antibacterial drugs will require a fundamental shift in how discovery through preclinical research is supported. PMID- 15757390 TI - Making dollars and sense of antisense. PMID- 15757391 TI - New advances in antibiotic development and discovery. AB - The development of new antibiotics is crucial to controlling current and future infectious diseases caused by antibiotic-resistant bacteria. Increased development costs, the difficulty in identifying new drug classes, unanticipated drug toxicities, the ease by which bacteria develop resistance to new antibiotics and the failure of many agents to address antibiotic resistance specifically, however, have all led to an overall decline in the number of antibiotics that are being introduced into clinical practice. Although there are few, if any, advances likely in the immediate future, there are agents in both clinical and preclinical development that can address some of the concerns of the infectious disease community. Many of these antibiotics will be tailored to specific infections caused by a relatively modest number of susceptible and resistant organisms. PMID- 15757392 TI - Agents and strategies in development for improved management of herpes simplex virus infection and disease. AB - The quiet pandemic of herpes simplex virus (HSV) infections has plagued humanity since ancient times, causing mucocutaneous infection such as herpes labialis and herpes genitalis. Disease symptoms often interfere with every-day activities and occasionally HSV infections are the cause of life-threatening or sight-impairing disease, especially in neonates and the immuno-compromised patient population. After infection the virus persists for life in neurons of the host in a latent form, periodically reactivating and often resulting in significant psychosocial distress for the patient. Currently no cure is available. So far, vaccines, ILs, IFNs, therapeutic proteins, antibodies, immunomodulators and small-molecule drugs with specific or non-specific modes of action lacked either efficacy or the required safety profile to replace the nucleosidic drugs acyclovir, valacyclovir, penciclovir and famciclovir as the first choice of treatment. The recently discovered inhibitors of the HSV helicase-primase are the most potent development candidates today. These antiviral agents act by a novel mechanism of action and display low resistance rates in vitro and superior efficacy in animal models. This review summarises the current therapeutic options, discusses the potential of preclinical or investigational drugs and provides an up-to-date interpretation of the challenge to establish novel treatments for herpes simplex disease. PMID- 15757393 TI - Investigational medications in the treatment of alopecia. AB - Androgenetic alopecia (hereditary thinning) is the most common cause of hair loss in both men and women. Chemotherapy-induced alopecia is another distressing cause of hair loss. With a better understanding of follicular biology and the signals responsible for hair growth and regression, targeted therapies for hair loss are being investigated. This review summarises investigational medications for androgenetic and chemotherapy-induced alopecia that are in preclinical stages or later. PMID- 15757394 TI - Cilansetron: a new serotonergic agent for the irritable bowel syndrome with diarrhoea. AB - Cilansetron is a novel serotonin type-3 (5-hydroxytryptamine; 5-HT) receptor subtype 3 (5-HT(3)) receptor antagonist currently being evaluated for the treatment of female and male patients with irritable bowel syndrome with diarrhoea predominance (IBS-D). 5-HT(3) receptor antagonists such as cilansetron have been shown to affect gastrointestinal motility. Whether cilansetron affects visceral sensation independent of effects on visceral compliance remains controversial. Results from two large, randomised, double-blind, placebo controlled, parallel-group Phase III clinical trials of cilansetron in patients with IBS-D have recently been presented in abstract form. These studies found that cilansetron was more effective than placebo at improving overall, as well as individual symptoms, including abdominal pain and diarrhoea in female and male IBS-D patients. The most commonly reported side effect with cilansetron has been constipation and, in general, the drug has been well tolerated in clinical trials. Although rare, the most concerning side effect observed with cilansetron has been suspected ischaemic colitis. The event rate for suspected ischaemic colitis associated with cilansetron from clinical trials is 3.77 per 1000 person years of exposure. This rate appears to be greater than that expected in the IBS population and similar to that observed with alosetron, another 5--HT(3) receptor antagonist. All of the cases of suspected ischaemic colitis reported with cilansetron have resolved without serious sequelae. How issues surrounding the safety of cilansetron will affect the approval process in various countries remains to be determined. However, the risk-benefit of cilansetron is likely to be most favourable in patients with IBS-D who have failed to respond to conventional medical therapies. A detailed risk management plan and post marketing surveillance programme will be required should this drug become available for the treatment of patients with IBS-D. PMID- 15757395 TI - Filling the hole: evidence of a small molecule binding to the fusion core pocket in human respiratory syncytial virus. PMID- 15757396 TI - A small-molecule lead compound for the treatment of Alzheimer's disease. PMID- 15757397 TI - 77th Scientific Sessions of the American Heart Association. AB - Nearly 4000 abstracts were selected for presentation at the 77th Scientific Sessions of the American Heart Association, held in New Orleans, Louisiana, USA. The sessions were divided into basic, clinical and population science. The abstracts have been published in a supplement to Circulation (2004) 110(7). PMID- 15757398 TI - Cambridge Healthtech Institute Signal Transduction Conference: targets for effective therapeutics. AB - The Cambridge Healthtech Institute Signal Transduction Conference covered three major topics over 2 days: the discovery of new signalling targets, improved technology to dissect out signal transduction pathways and the effects of small molecules on those pathways, and progress in the discovery and development of signalling modulators. There was a particular emphasis placed on the biology of protein kinases and industry efforts to develop efficacious and safe inhibitors for this target class. Of note, kinase inhibitors for therapeutic indications other than oncology, including those directed against inflammation, allergy and metabolic disorders, have reached or have nearly completed clinical testing. Other signalling targets presented included tyrosine phosphatases, intracellular and membrane-bound channels, and G-protein-coupled receptors. This article will briefly summarise the newer technologies and signalling targets but will primarily focus on the presentations covering drug discovery and development. PMID- 15757399 TI - Future of allergen-specific immunotherapy. AB - Allergen-specific immunotherapy (SIT) has been used for almost a century as a desensitising therapy for allergic diseases. Administration of appropriate concentrations of allergen extracts has been shown to be reproducibly effective when patients are carefully selected. The disadvantage with allergen extracts is that they consist of nonallergenic or even toxic proteins, and can induce severe side effects including anaphylaxis and death. Several strategies have been developed to tackle this issue. With the introduction of recombinant DNA technology and peptide chemistry, it became possible to produce SIT vaccines with reduced allergenic activity. In addition, current understanding of immunological mechanisms of SIT, particularly the role of regulatory T cells in allergen specific peripheral tolerance, may enable novel treatment strategies. PMID- 15757400 TI - Emerging human papillomavirus vaccines. AB - Human papillomavirus (HPV) infections are a leading cause of virus-associated cancers of the anogenital, oropharyneal and cutaneous epithelium. The most prevalent of these is cervical cancer, which is responsible for approximately 500,000 deaths annually worldwide. A group of about 15 serologically unrelated 'high-risk' HPV types are responsible for almost all HPV-associated cancers. Prevention of papillomavirus infection can be achieved by induction of capsid specific neutralising antibodies in preclinical animal papillomavirus models and in recent human clinical trials. High titres of conformationally-dependent, type specific HPV-neutralising antibodies are triggered by HPV virus-like particle (VLP) vaccines. Overcoming the problems of type-specificity of the responses to these VLP vaccines is a potentially important area of current HPV vaccine research, with an emphasis on induction of more broadly cross-protective neutralising responses. Viral oncogenes E6 and E7 are continuously present in HPV associated cancers and are prime targets for HPV therapeutic vaccines. A variety of approaches are being tested in therapeutic vaccine clinical trials and in various preclinical animal papillomavirus models for efficacy. Approaches include genetic vaccines, recombinant virus vaccines, dendritic cell-based strategies, immunomodulatory strategies and various combination strategies to maximise cell mediated immunity to papillomavirus proteins present in HPV infections and cancers. The success of preventive HPV VLP vaccines in clinical trials is clear. However, current therapeutic vaccine trials are less effective with respect to disease clearance. Nevertheless, a series of combination approaches have shown significant therapeutic enhancement in preclinical papillomavirus models and await testing in patient populations to determine the most effective strategy. There is much encouragement that HPV vaccines will be the most effective approach to prevention and cure of infections caused by this group of viruses, which re present a significant human pathogen. PMID- 15757401 TI - Emerging azole antifungals. AB - Systemic and superficial fungal infections have progressively emerged over the past few decades as an increasing cause of human disease, especially in the immunocompromised host. Control of fungal disease has proved difficult because few risk factors are potentially preventable; the population at highest risk for fungal disease, the immunocompromised patient, has been steadily increasing. There is a clear need for additional safe and effective therapeutic agents for the treatment of systemic fungal disease. A new generation of triazoles that includes voriconazole, posaconazole, ravuconazole and albaconazole has emerged and are presently in different phases of clinical investigation. These new triazoles have demonstrated a broad spectrum of activity, in particular against fungal pathogens previously resistant to previously available antifungals. This review highlights the emerging azole antifungals, both those available and in clinical development, and discusses their prospects for the future. PMID- 15757402 TI - Emerging drugs for moderate-to-severe psoriasis. AB - Psoriasis is a chronic, incurable, disabling skin disease characterised by red, scaly plaques. Approximately 23% of psoriasis patients also have an accompanying arthritis that can become debilitating. Psoriasis has a stigmatising effect on its victims, who often feel socially isolated. Although the exact aetiology of psoriasis is still unknown, it is clearly an immune-mediated disease. Traditional therapies for psoriasis include topical drugs, such as corticosteroids, retinoids and vitamin D3 analogues; systemic drugs, such as methotrexate, ciclosporin and retinoids; and phototherapy. These mainstays of treatment are efficacious for the treatment of severe disease; however, most are associated with toxicities or are inconvenient. Recent advances in biotechnology have produced new pharmaceuticals that interfere with immune responses thought to be involved in the pathogenesis of psoriasis and other inflammatory diseases. The immunobiologicals, one new family of drugs, consist of monoclonal antibodies and fusion proteins. Many have demonstrated efficacy in treating psoriasis. Some appear to offer safety benefits over traditional therapies; further monitoring and surveillance of these agents is required to adequately establish safety profiles. This article discusses existing and emerging treatments for moderate-to-severe psoriasis. PMID- 15757403 TI - Emergence of targeted immune therapies for systemic lupus. AB - Systemic lupus erythematosus (SLE) is a prototypical autoimmune disease, characterised by flares of rampant inflammation that can threaten, in an unpredictable manner, almost any organ in the body. Current standard of care is largely empiric, involving the use of corticosteroids and toxic immune suppressive agents that are widely acknowledged to have unacceptable side effects for long-term use. Recently, there have been significant advances in understanding the nature of some fundamental immune imbalances underlying the complicated clinical manifestations of SLE. Nevertheless attempts to develop and test more targeted, and potentially safer immune-modulating drugs for lupus have encountered significant obstacles, due to the lack of validated biological markers for disease flare and remission, and difficulties in the clinical assessment of the heterogeneous patients. In support of renewed interest in drug development for lupus, large collaborative groups have formed, and efforts are underway to develop objective biomarkers for SLE as well as to improve the standardisation and reproducibility of clinical outcome measures in multi-centre trials. PMID- 15757404 TI - The drug-based pipeline against restenosis. AB - More than 1 million percutaneous coronary interventions (PCIs) are performed yearly worldwide. Restenosis is the recurrent narrowing that can occur within 6 months following an initially successful PCI. Although drug-eluting stents have accomplished remarkable success, restenosis has not been eliminated and optimisation of both the polymers and drugs associated with them is desirable. This article reviews the presently available and potential preventive approaches against restenosis, including the sirolimus and paclitaxel drug-eluting stents. PMID- 15757405 TI - Emerging strategies for treatment of venous thromboembolism. AB - Although considerable progress has been made in the treatment of venous thromboembolism (VTE), many unanswered questions remain, which are awaiting proper solution. Furthermore, new opportunities are emerging, which have the potential to rapidly change the therapeutic scenario. Selected patients with deep vein thrombosis can be effectively and safely treated at home with fixed-dose low molecular-weight heparins. The long-term use of low-molecular-weight heparins is likely to be more effective than and as safe as oral anticoagulants for the secondary prevention of VTE in cancer patients with venous thrombosis. Recent publications have unexpectedly raised a renewed interest on the use of thrombolytic drugs in patients with pulmonary embolism, at least in those who present with heart ventricular dysfunction. The optimal long-term treatment of VTE is still undefined. Finally, new categories of drugs are emerging, which have the potential to replace conventional anticoagulants in the near future. They include anti-Xa inhibitors, such as pentasaccharide, and antithrombin inhibitors, such as ximelagatran. PMID- 15757406 TI - Emerging therapies for neuropathic pain. AB - Neuropathic pain develops as a result of damage to either the peripheral or central nervous system. It is characterised by spontaneous burning pain and/or ongoing pain with accompanying hyperalgesia and allodynia. Neuropathic pain is difficult to treat as it is often refractory to conventional analgesic treatments, with most patients obtaining only partial relief. At present, there are four major medication categories that are considered first-line treatment for neuropathic pain: antidepressants, anticonvulsants, local anaesthetic/topical agents and opioids. The efficacy of these treatments in neuropathic pain, excepting opioids, has been discovered serendipitously. However, responder rates and overall efficacy is poor with these agents and tolerability or side effects are often limiting. This update will review existing treatment options for neuropathic pain, and highlight more recent advances in the development of novel analgesics to treat this chronic disorder. PMID- 15757407 TI - Current and emerging medical therapies for glaucoma. AB - Glaucoma is a multifactorial optic neuropathy in which there is a characteristic acquired loss of retinal ganglion cells, at levels beyond normal age-related baseline loss, and corresponding atrophy of the optic nerve. Although asymptomatic in its earlier stages, the disease is nevertheless one of the leading global causes of irreversible blindness. Although elevated intraocular pressure (IOP) is one of the most important risk factors and lowering of IOP is the only proven treatment so far, the definition of glaucoma has evolved from a disease caused by increased IOP to one characterised by an IOP-sensitive, progressive optic neuropathy. In recent years, safer and better tolerated topical medications have been developed to control IOP more effectively, thereby limiting the need for surgery. New research has also noted the importance of diurnal IOP variation as a critical risk factor for progression of glaucomatous optic neuropathy (GON) and subsequent visual field loss. Moreover, new discoveries have further elucidated the basic pathophysiological and genetic mechanisms underlying the elevated levels of IOP, as well as the cellular mechanisms of GON. As our understanding of these complex pathways continues to improve, development opportunities for new therapeutic modalities will be enhanced. PMID- 15757408 TI - Future pharmacological treatment options for nonexudative and exudative age related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss in the industrialised world. Within the past decade, researchers have introduced many promising prevention and treatment options in an attempt to minimise the central vision loss imparted from AMD. Based on large-scale, randomised, prospective, placebo-controlled trials, a specially formulated combination of the antioxidants vitamin C, vitamin E, beta-carotene, copper and zinc is the only proven means of AMD prophylaxis. Thermal laser photocoagulation and photodynamic therapy with verteporfin are the only standard treatment options. However, efficacy is limited and treatment is only applicable to a minority of AMD patients. Thus, alternative pharmacological interventions are in all phases of clinical development. Researchers are guardedly optimistic that these advances may change the entire approach to AMD management in the near future. This review article will detail the currently accepted treatment options, as well as describe several of the more promising investigational pharmacological approaches to AMD. PMID- 15757409 TI - Heat-shock protein 90 inhibitors as novel cancer chemotherapeutics - an update. AB - Heat-shock protein 90 (Hsp90) is a molecular chaperone whose association is required for stability and function of a growing number of signalling proteins that have been implicated in cancer cell survival, including several mutated proteins that are only found in specific cancers. Furthermore, a growing body of evidence suggests that cancer cells are particularly dependent on Hsp90 for their growth and survival, and, therefore, are more sensitive to the effects of its inhibition than are non-transformed cells and tissues. Several chemically distinct Hsp90 inhibitors have shown encouraging antitumour activity in multiple preclinical model systems, and one Hsp90 inhibitor, the benzoquinone ansamycin 17 allylamino, 17-demethoxygeldanamycin, has completed five Phase I clinical trials, with a number of Phase II trials soon to be underway or in progress. Other Hsp90 inhibitors are either in Phase I clinical trial or under development. This update will focus on how the latest developments in Hsp90 biology may better inform the clinical development of Hsp90 inhibitors. PMID- 15757410 TI - Emerging analgesics in cancer pain management. AB - Cancer pain is one of the most frequent symptoms in malignant disease, severely impairing the patients' quality of life. The recommendations of the World Health Organization will provide adequate pain relief for the vast majority of cancer patients. However, some patients will suffer from inadequate analgesia or intolerable side effects. Cyclooxygenase-2 (COX-2)-selective non-steroidal anti inflammatory drugs (NSAIDs), new anticonvulsants, cannabinoids and NMDA receptor antagonists are being developed for these patients. NSAIDs with nitric oxide releasing moieties are an interesting addition, as this new class of analgesics combines improved analgesic efficacy with higher tolerability. Conotoxins and other drugs such as nicotinic acetylcholinergic receptor agonists will be advantageous only for a few patients in the near future, as side-effect profile and risk of complications, as well as the burden on the patient, often are not worth the additional analgesic benefit. PMID- 15757411 TI - Therapy for haemophilia: recent advances and goals for the future. AB - In the 20th century, haemophilia evolved from a life-threatening, crippling disease to one for which the prognosis is excellent and many patients lead normal, productive lives. Although dramatic achievements in the treatment of haemophilia have occurred, the current therapies have significant drawbacks. Among these is the relatively high incidence of inhibitor development, the requirement for frequent intravenous infusions to prevent bleeding complications, the lack of effective treatment for established joint disease, and the high cost of treatment. The future goal of haemophilia treatment first and foremost is curing this genetic condition via gene therapy. As this goal is likely many years away, improvements in the current factor products in order to reduce the development of inhibitors and to reduce the frequency of therapy are more immediately achievable goals. Finally, improving the treatment of bleeding complications, particularly in inhibitor patients, and developing novel adjunctive therapies for the management of joint disease are also important goals for the near future. This review will discuss in detail the cu-rrent and future goals of haemophilia therapy. PMID- 15757412 TI - Transgenic plants in the biopharmaceutical market. AB - Many of our 'small-molecule-drugs' are natural products from plants, or are synthetic compounds based on molecules found naturally in plants. However, the vast majority of the protein therapeutics (or biopharmaceuticals) we use are from animal or human sources, and are produced commercially in microbial or mammalian bioreactor systems. Over the last few years, it has become clear that plants have great potential for the production of human proteins and other protein-based therapeutic entities. Plants offer the prospect of inexpensive biopharmaceutical production without sacrificing product quality or safety, and following the success of several plant-derived technical proteins, the first therapeutic products are now approaching the market. In this review, the different plant based production systems are discussed and the merits of transgenic plants are evaluated compared with other platforms. A detailed discussion is provided of the development issues that remain to be addressed before plants become an acceptable mainstream production technology. The many different proteins that have already been produced using plants are described, and a sketch of the current market and the activities of the key players is provided. Despite the currently unclear regulatory framework and general industry inertia, the benefits of plant-derived pharmaceuticals are now bringing the prospect of inexpensive veterinary and human medicines closer than ever before. PMID- 15757413 TI - Inflammation Research Association: 12th international conference. AB - The Inflammation Research Association held its 12th international meeting at the Sagamore at Bolton Landing in New York State (3 - 7 October 2004). These meetings were originally intended for scientists from the pharmaceutical industry to get together and discuss the latest developments in inflammation drug discovery, and it remains an industry-dominated affair. The conference covered some highly topical issues such as cyclooxygenase-2 inhibitors (rofecoxib/Vioxx [Merck & Co., Inc.] was withdrawn from the market only a few days before the conference), along with areas of ongoing interest to the pharmaceutical and biotechnology industry, including p38 MAPK inhibitors, nuclear hormone receptor modulators and prostaglandin receptor antagonists. This review will cover the main themes that emerged during the meeting. PMID- 15757414 TI - Hepatitis A vaccines: the growing case for universal immunisation of children. AB - Hepatitis A virus (HAV) infection results in a debilitating, multi-week illness in older children and adults that can be avoided with pre-exposure immunisation. Inactivated HAV vaccines are safe, highly effective and induce long-lasting protection. Immunisation has typically been recommended only for groups at highest risk of infection or complications, with variable success. This paper reviews the outcomes of several new programmes of universal HAV immunisation of children and/or adolescents. These programmes were associated with substantial disease rate reductions, extending beyond the immunised individuals and age groups (herd immunity), and promise additional benefits as immunised cohorts enter adulthood risk groups. Universal programmes are cost-effective, especially when a combined hepatitis A and B vaccine can be used to minimise incremental costs, and deserve wider consideration in developed countries. PMID- 15757415 TI - Pityriasis versicolor: a review of pharmacological treatment options. AB - Pityriasis versicolor is a common disorder of the skin, which is characterised by scaly hypo- or hyperpigmented lesions on the body. The lipophilic yeast, Malassezia, is considered to be the aetiological agent of this disease. A number of treatment options, both topical and systemic, have been shown to be effective. A critical evaluation of treatment options is presented. PMID- 15757416 TI - Treatment strategies underpinning the global programme to eliminate lymphatic filariasis. AB - Lymphatic filariasis (LF) is a disease targeted for elimination. The global strategy is a once-yearly, single-dose, two-drug regimen utilised by communities at risk for LF, with the goal of reaching 80% population coverage yearly, for at least 5 years, in order to interrupt transmission of LF. Where onchocerciasis is co-endemic, the regimen is ivermectin 200 - 400 microg/kg plus albendazole 400mg; elsewhere, the regimen should be diethylcarbamazine 6 mg/kg plus albendazole 400mg. This paper reviews in detail the evidence for the efficacy and safety of these two-drug regimens underpinning the global strategy and makes recommendations for future developments in chemotherapy for LF, focusing on unresolved issues. These include optimal frequency, duration and end point of treatment, tools for monitoring successful therapy and means for detecting the potential development of resistance to any of the three antifilarial drugs on which the Global Programme to Eliminate LF depends. PMID- 15757417 TI - Effective treatments in the management of brucellosis. AB - Treatment of uncomplicated brucellosis in humans utilises a variety of anti biotic combinations, applied to a series of important pathogenetic and clinical parameters. The currently recommended treatment regimens have not been surpassed by newer compounds, and various therapeutic strategies utilising these compounds cannot be adequately evaluated due to the absence of large, multi-centre, multinational trials. The review focuses on the basic principles of brucellosis treatment, the properties of the various regimens used, the results of trials involving them, and the questions raised about the efficacy of these regimens in certain clinical situations. PMID- 15757418 TI - Biliary tract cancers: current concepts and controversies. AB - Biliary tract cancer, which consists of gall bladder cancer and cholangio carcinoma, presents many challenges to practising physicians. It is a relatively rare cancer that often causes a diagnostic dilemma, as its presentation may be similar to that of non-malignant conditions. In many cases, histological or cytological confirmation of a cancer diagnosis is not possible preoperatively. The management of this disease is also complex due to a morbid patient population and limited data on the optimal therapeutic approach. Surgery remains the mainstay of treatment, although the extent of resection required is still debated. The role of adjuvant therapy is also controversial, but a combined modality approach appears to be beneficial in patients with a high risk of recurrence, such as those with node positive tumors or positive resection margins. When surgery is not possible, the prognosis of patients with biliary tract cancer is very poor. In unresectable patients, the combination of chemotherapy and radiotherapy can result in a prolonged survival for some patients. In the palliative setting, biliary stenting and other supportive measures can alleviate symptoms and improve survival. Gemcitabine-based combination chemotherapy may also provide successful palliation and has achieved response rates of approximately 30% and a median survival of > 15 months in one study. Ultimately, treatment decisions should be individualised and participation in clinical trials is encouraged. Further progress in the management of biliary tract cancer is anticipated using biological therapies and continued research is essential to discover the optimal treatment for this challenging disease. PMID- 15757419 TI - Available options in chemotherapy for advanced gastric cancer: the current developments in Japan. AB - Although techniques for early diagnosis and treatment of gastric cancer have been established, standard treatment for advanced gastric cancer has not. 5 Flourouracil (5-FU) plus cisplatin containing regimens, including 5-FU plus cisplatin, epirubicin plus cisplatin and 5-FU, and docetaxel plus cisplatin and 5 FU, are considered to be the most effective regimens for advanced gastric cancer in US, Europe and Korea. In Japan, oral fluoropyrimidine S-1 is currently considered to be the first-line treatment for advanced gastric cancer. S-1-based combination therapies with other promising drugs, including cis-platin, irinotecan and taxanes, are expected to yield good results. Above all, S-1 plus cisplatin therapy showed high efficacy and expected to be a standard therapy for advanced gastric cancer. Further evaluation by well-controlled clinical trials is still required. PMID- 15757420 TI - Drug interactions in the management of HIV infection. AB - The availability of antiretroviral therapy has significantly reduced the morbidity and mortality of HIV infection. In addition, improved treatment of opportunistic infections and comorbidities common to patients with HIV is further prolonging the lives of patients. Improvement in the treatment of HIV has led to a significant increase in the number of medications which caregivers are able to utilise to manage HIV/AIDS. Antiretroviral medications, as well as many of the drugs used in the management of opportunistic infections and primary care (e.g., macrolide antibiotics, azole antifungals, cholesterol-lowering medications), are particularly prone to drug interactions. The interpretation of clinically significant interactions is complicated by the rate at which new information on drug metabolism and transport is becoming available. Management of drug interactions in HIV is further confounded by conflicting study results and differences between documented and theoretical inter-actions. The mechanisms and significance of interactions involving antiretrovirals, drugs used for opportunistic infections, and other medications commonly used in HIV patients will be reviewed. PMID- 15757421 TI - Current pharmacological options in the treatment of croup. AB - Croup is one of the most common respiratory illnesses seen in the acute paediatric setting. It can be a cause of acute stridor and/or respiratory distress in young children. Research has shown that therapy aimed at reducing symptoms and inflammation can reduce complications such as the need for intubation, hospitalisation and improve quality of life for parents and children. Corticosteroids are the primary treatment option that will accomplish both goals and can be used in out-patient and in-patient settings. Corticosteroids may be given orally, parenterally or in wet nebulised form; however, oral administration is the preferred route. Wet nebulised adrenaline (racaemic or l-adrenaline) is also an effective treatment for more severe cases of croup. Recent studies have shown that mist/humidified air provides no additional symptom improvement, nor does it alter the overall cause of the disease process. Currently, there is insufficient randomised controlled trial evidence to support the role of heliox in the short-term treatment of croup. PMID- 15757422 TI - Antimicrobial interventions for the management of diabetic foot infections. AB - Foot infections are the most common cause of hospitalisations and amputations in diabetic patients. They occur after skin ulcers or trauma in patients with peripheral neuropathy, sometimes together with vascular disease. Narrow-spectrum antibiotic agents should be prescribed for minor recent infections, and broader spectrum agents for severe or chronic infections. When indicated, antibiotic therapy should be started early and be tailored to the individual patient. Diabetic foot osteomyelitis is a particularly controversial condition, especially regarding the need for reliable cultures, the type and duration of treatment, and the role of surgery. Recent data indicates that a medical approach might be effective and could reduce foot amputations among diabetic patients. Interdisciplinary cooperation with infectious disease specialists and orthopaedic surgeons should be considered in such situations. PMID- 15757423 TI - The Avoiding Cardiovascular events through COMbination therapy in Patients Living with Systolic Hypertension (ACCOMPLISH) trial: a comparison of first-line combination therapies. AB - Although multidrug therapy is required in order to achieve good blood pressure control in many hypertensives, there are no studies directly comparing fixed-dose combinations as initial therapy. The Avoiding Cardiovascular events through COMbination therapy in Patients Living with Systolic Hypertension (ACCOMPLISH) trial compares regimens of benazepril plus amlodipine versus benazepril plus hydrochlorothiazide, force-titrated to 40/10 and 40/25mg, respectively. A total of 12,600 high-risk hypertensives have been randomised and will be followed for 3 - 5years, during which cardiovascular events will be monitored. The investigators hypothesise that the benazepril plus amlodipine regimen will decrease cardiovascular events by 15% compared with benazepril plus hydrochlorothiazide. Recruitment began in 2003, and the trial is expected to end in 2008. The ACCOMPLISH trial shares important limitations with many other recent trials that will make it difficult to apply the results in clinical practice. These include the focus on high-risk hypertensive patients, in whom significant reductions in relative risk will translate into meaningful reductions in absolute risk: in lower-risk hypertensives with a low absolute risk, similar relative risk reductions may not be of great impact on the population disease burden. In ACCOMPLISH, as in most industry-sponsored clinical trials, the main goal appears to be market-driven: doses of drugs tested are not those available for clinical practice. The question asked, whether the combination of benazepril with either diuretic or dihydropyridine calcium channel blocker is more efficacious, is not a clinically compelling one. Finally, the univariate subgroup analyses proposed are unlikely to lead to an understanding of whether either combination has specific advantages for patients encountered clinically, most of whom have multiple risk factors. Thus, it appears that ACCOMPLISH, as with many recent pharmacological trials, will not greatly impact the treatment of hypertension. PMID- 15757424 TI - Moxifloxacin in respiratory tract infections. AB - Moxifloxacin is a fourth-generation fluoroquinolone that has been shown to be effective against respiratory pathogens, including Gram-positive (Streptococcus pneumoniae), Gram-negative (Haemophilus influenzae, Moraxella catarrhalis), and atypical strains (Chlamydia pneumoniae, Mycoplasma pneumoniae), as well as multi drug resistant S. pneumoniae, including strains resistant to penicillin, macrolides, tetracyclines, trimethoprim/sulfamethoxazole and some fluoroquinolones. Moxifloxacin is highly concentrated in lung tissue, and has demonstrated rapid eradication rates. The bioavailability and half-life of moxifloxacin provides potent bactericidal effects at a dose of 400mg/day. The ratio of the area under the concentration-time curve to MIC of moxifloxacin is the highest among the fluoroquinolones against S. pneumoniae. The clinical efficacy of moxifloxacin has been shown in controlled studies of community acquired pneumonia (CAP), exacerbations of chronic bronchitis (CB) and acute bacterial rhinosinusitis. Moxifloxacin has demonstrated a faster resolution of symptoms in CAP and exacerbations of CB patients compared with first-line therapy. It has also demonstrated better eradication in exacerbations of CB compared with standard therapy, in particular the macrolides. Treatment guidelines should take into account the results of clinical trials with moxifloxacin in order to establish the role of this antimicrobial in the therapeutic arsenal against respiratory tract infections. PMID- 15757425 TI - Epoetin alfa for the treatment of cancer- and chemotherapy-related anaemia: product review and update. AB - Anaemia, often associated with chemotherapy, is a common and debilitating disorder in cancer patients. Recombinant human erythropoietin (epoetin alfa) was introduced in the 1990s for the treatment of chemotherapy-related anaemia. Data from randomised, double-blind, placebo-controlled studies and large, non randomised, community-based studies have demonstrated that either of the FDA approved dosing schedules of epoetin alfa 150 - 300 U/kg three times weekly or 40,000-60,000 U/week s.c., significantly increases haemoglobin levels, reduces transfusion requirements, and improves quality of life in anaemic cancer patients undergoing chemotherapy or chemoradiation therapy. Guidelines for the effective and safe use of epoetin alfa have been published by major oncology/haematology organisations and are reviewed in this article. Areas of recent and ongoing investigation with epoetin alfa are also covered in this review. PMID- 15757426 TI - Dutasteride: a novel dual inhibitor of 5alpha-reductase for benign prostatic hyperplasia. AB - Dutasteride is a new dual 5alpha-reductase inhibitor for the treatment of benign prostatic hyperplasia. It differs from finasteride as it inhibits both isoenzymes of 5alpha-reductase and results in near-complete suppression of serum dihydro testosterone. Similar to finasteride, it reduces serum prostatic specific antigen by approximately 50% at 6months and total prostate volume by 25% in 2years. Randomised, placebo-controlled trials conducted over 2years have shown the efficacy of dutasteride in symptomatic relief, improvements in quality of life and peak urinary flow rate, and reduction of acute urinary retention events and the need for surgery. The main side effects are erectile dysfunction, decreased libido, gynaecomastia and ejaculation disorders. However, long-term usage for > 4years did not reveal increased new onset of sexual side effects. In addition, the combination of dutasteride and tamsulosin is well-tolerated and has the added advantage of rapid symptomatic relief. Finally, dutasteride has been shown to possess tumour regression properties invitro and its role in chemoprevention of prostate cancer will be confirmed in the ongoing Reduction by Dutasteride of Prostate Cancer Events (REDUCE) trial. PMID- 15757427 TI - Lanthanum carbonate for the treatment of hyperphosphataemia in renal failure and dialysis patients. AB - Hyperphosphataemia is a usual accompaniment of end stage renal disease and dialysis, in the absence of dietary phosphate restriction or supplemental phosphate binders. It is associated with renal osteodystrophy, metastatic calcification and increased mortality and morbidity. Despite dietary restriction and dialysis, most patients will require a phosphate-binding agent to treat this condition. However, phosphate control has not significantly improved over the last two decades, mainly because of the lack of an ideal oral phosphate-binding agent. Aluminium- and calcium-based agents are associated with major side effects, despite their undoubted efficacy. Although sevel-amer hydrochloride represents a step forward in the management of hyperphosphataemia, it is not an ideal phosphate binder due to its cost and tablet burden. Lanthanum carbonate is the most recent non-calcium, non-aluminium, phosphate-binding agent. It is effective and well-tolerated, and no negative effects on bone histology have been observed. PMID- 15757428 TI - Sodium oxybate for the treatment of narcolepsy. AB - Narcolepsy, a lifelong disease with diverse symptoms, poses a therapeutic challenge to physicians. Psychomotor stimulants are used to provide some relief from excessive sleepiness, whereas a variety of other medications have traditionally been used to treat the other symptoms of this disorder. Cataplexy, consisting of sudden episodes of bilateral skeletal muscle weakness, has long been treated with tricyclic antidepressants or selective serotonin re-uptake inhibitors. Although these drugs have brought relief to some patients, they cause intolerable adverse effects in others, whereas still others become tolerant to their beneficial effects. In July of 2002, sodium oxybate was approved by the US FDA for the treatment of cataplexy, representing a significant advance in the treatment of this unusual disease. The following drug evaluation summarises the role of this novel medication in the practice of sleep medicine. PMID- 15757429 TI - Ziprasidone: efficacy, tolerability, and emerging data on wide-ranging effectiveness. AB - Ziprasidone is the fifth atypical antipsychotic to become available in the US market. This compound is a serotonin-dopamine-receptor antagonist, with greater affinity for the 5-HT(2A) receptor than the dopamine D2 receptor. Similar to aripiprazole, this compound also has agonist activity at the serotonin 5-HT(1A) receptor. This affinity has the potential to have particularly beneficial effects on cognitive and affective abnormalities in schizophrenia. Oral and short-acting intramuscular formulations are available. This compound has recently been approved for the treatment of acute mania as well as for schizophrenia. Ziprasidone is associated with low weight gain and a low potential to change lipid and glucose levels. Long-term tolerability data have indicated that initial concerns regarding QTc (corrected cardiac output) prolongation are not a major issue. Recent data have indicated that this compound has short-term antipsychotic efficacy that is equivalent to other members of the class, also with evidence of cognitive enhancement. Finally, longer term data has indicated that ziprasidone is successful in the prevention of relapse and has sustained cognitive benefits. Several issues remain to be addressed, including the efficacy of once-daily dosing, the need to take the medication with food, and the correct dosing strategy. PMID- 15757430 TI - The safety of lumiracoxib when used in the treatment of arthritis. PMID- 15757431 TI - The ODAC Chronicles: Part 3. The FDA's philosophy and process for cancer drug evaluation and approval. PMID- 15757432 TI - Robotic surgery: the coming of a new era in surgical innovation. PMID- 15757434 TI - Docetaxel in advanced non-small cell lung cancer. AB - Based on the survival benefit demonstrated in large randomized clinical trials, docetaxel is approved for the treatment of advanced non-small cell lung cancer (NSCLC) in both the first- and second-line settings. The efficacy of docetaxel in combination with cisplatin is equivalent to some, and superior to other, platinum based doublets for first-line management of NSCLC, and has a manageable toxicity profile. Carboplatin-based regimens and nonplatinum combinations with docetaxel also have proven efficacy in first-line therapy of patients with advanced NSCLC. Combinations of docetaxel with various novel targeted agents have produced encouraging data in Phase II studies. This article reviews recent studies of docetaxel as a single agent and in combination regimens with cytotoxic and more recent targeted agents in the management of advanced NSCLC. PMID- 15757435 TI - Thalidomide: present and future in multiple myeloma. AB - Multiple myeloma continues to be an incurable disease. The understanding of the disease's pathophysiology has significantly improved over the past few years, partly due to the discovery of the role of immunomodulatory agents and the study of their mechanism of action. Thalidomide, the first of the immunomodulatory family to be used in the management of multiple myeloma, proved not only to be effective in the treatment of multiple myeloma, but also instigated a wide range of in vitro and in vivo studies to define the pathophysiology of the plasma cell dyscrasia. The attention thalidomide has received in the past and recent history has not been without a price. The drug has a side-effect profile that, if managed appropriately, provides the most unique active molecule in the management of the disease, where it maintains the same response rate in newly diagnosed patients as in advanced relapsed/refractory multiple myeloma patients. PMID- 15757436 TI - Denileukin diftitox: a concise clinical review. AB - Denileukin diftitox (DAB389IL-2; Ontak) is a novel recombinant fusion protein approved by the US Food and Drug Administration for the treatment of relapsed or refractory cutaneous T-cell lymphoma. It consists of fragments of diphtheria toxin linked to human interleukin-2 and works by targeting the high-affinity interleukin-2 receptor expressed on malignant cells. This article will review the clinical trials leading to the approval of denileukin diftitox for cutaneous T cell lymphoma, and discuss the potential future role of this novel drug in patients with both malignant and nonmalignant diseases, including non-Hodgkin's lymphoma, chronic lymphocytic leukemia, solid tumors, psoriasis and graft-versus host disease. PMID- 15757437 TI - Alemtuzumab. AB - Alemtuzumab is a humanized monoclonal antibody against CD52, a small glycosylphosphatidylinositol-anchored glycoprotein that is highly expressed on normal T- and B-lymphocytes, and on a large proportion of malignant lymphoid cells, but not on hematopoietic progenitor cells. Over the past several years, a number of clinical trials have demonstrated the clinical activity of alemtuzumab in treating patients with chronic lymphocytic leukemia, T-cell malignancies such as T-prolymphocytic leukemia and cutaneous T-cell lymphoma, as well as in the prevention and therapy of graft-versus-host disease in the setting of allogeneic stem cell transplantation. Its application in a number of autoimmune disorders is currently under investigation. The most significant side effect of alemtuzumab is predisposition to infections related to the associated profound lymphopenia. Despite this, and with appropriate and more effective antibiotic prophylaxis, it is likely that we will witness an expansion of the role of alemtuzumab in the future. PMID- 15757438 TI - New standards in the chemotherapy of metastatic hormone-refractory prostate cancer. AB - Hormone-refractory prostate cancer (HRPC) is a major issue in Western countries and the second leading cause of cancer death in North American men. In the prostate-specific antigen era, most HRPCs are currently diagnosed in asymptomatic patients based on biochemical failure, with increasing demand for active treatment. Until recently, chemotherapy for HRPC patients was not considered a standard of care due to the absence of clear data evidencing an overall survival benefit. In fact, few Phase III studies conducted in the 1980s and early 1990s had documented a superiority over corticosteroids alone in terms of biochemical response (declines in serum prostate-specific antigen levels) and quality of life, but not survival. Due to their impact on pain control, mitoxantrone and prednisone were long considered the best regimen for symptomatic HRPC patients. In recent years, more chemotherapeutic agents have been tested, among which the microtubule inhibitors (vinca alkaloids and taxanes) have obtained the most promising results in Phase II trials and have entered Phase III testing. Two well designed randomized trials have changed this scenario. Both compared docetaxel (with or without estramustine) against mitoxantrone and prednisone, and demonstrated a significant advantage not only in terms of response, pain control and quality of life, but also in terms of overall survival. Which patients need to be treated, the regimen of choice and duration of chemotherapy will be the next questions to be answered in the coming years in the field of HRPC, along with the role of new signal transduction inhibitors and other targeted therapies. PMID- 15757439 TI - Regulation of androgen receptor signaling in prostate cancer. AB - Prostate cancer is a significant cause of morbidity and mortality worldwide. Normal prostate tissue is regulated by androgens, which activate the androgen receptor, a nuclear receptor transcription factor. Most prostate tumors retain androgen dependence, therefore, current therapies for advanced prostate cancer either reduce androgen levels or prevent binding to the androgen receptor. Despite this regimen, prostate cancer invariably progresses to a fatal, androgen refractory state. Although these relapsed tumors are androgen independent, they are still dependent on the androgen receptor for their growth and survival. The focus of this review will be to highlight our current understanding of the mechanisms of androgen receptor activation in androgen-refractory prostate cancer. How these mechanisms of androgen receptor activation could be targeted in this advanced stage of the disease is also discussed. PMID- 15757440 TI - Retroperitoneal lymph node dissection in the treatment of low-stage nonseminomatous germ cell tumors of the testicle. AB - Nonseminomatous germ cell tumors of the testicle are highly treatable and curable. The evolution of cancer control for this disease has demonstrated an effective integration of medical and surgical approaches over the last 30 years. Current emphasis in the therapy of nonseminomatous germ cell tumors focuses on minimizing treatment-related morbidity while maintaining consistently high cure rates. Retroperitoneal lymph node dissection in experienced hands is a critical component of the treatment armamentarium in this disease. Retroperitoneal lymph node dissection is an accurate staging tool that provides important information in determining the need for chemotherapy. When performed properly, retroperitoneal lymph node dissection eliminates the retroperitoneum as a site for relapse, which in turn provides emotional and psychological relief to the patient and simplifies the follow-up protocol. Retroperitoneal lymph node dissection alone can also provide high cure rates in patients with clinical low stage disease and high-risk factors such as lymphovascular invasion or predominance of embryonal histology in the primary tumor. Teratoma is known to be chemoresistant and, when present in the primary tumor of low-stage patients, may be best treated with primary retroperitoneal lymph node dissection. Primary chemotherapy in the treatment of low-stage nonseminomatous germ cell tumors deserves continual investigation as long-term toxicities become more apparent. Observation is an option for the highly motivated patient, but requires a rigorous follow-up schedule to avoid relapse. Laparoscopic retroperitoneal lymph node dissection is a viable staging tool, however, oncologic control of the retroperitoneum has not been reliably determined. PMID- 15757441 TI - Monoclonal antibody therapy of ovarian cancer. AB - Despite advances in understanding and treatment, ovarian cancer remains a major cause of cancer mortality worldwide. Debulking surgery and paclitaxel/carboplatin chemotherapy induce good initial responses in most patients, although most cases of advanced disease are not controlled. Monoclonal antibodies hold promise as a potential incremental advance for the treatment of the disease. Antibodies can be used to stimulate the immune response, target tumor-specific receptors to induce antibody-dependent cellular cytotoxicity or interfere with biologic pathways. They can also be used to deliver therapeutic radioisotopes to malignant cells. Oregovomab is in Phase III clinical trials as a consolidation treatment post front-line therapy to trigger tumor-specific cellular immunity. Bevacizumab, which blocks vascular endothelial growth factor, will be entering Phase III as an adjuvant to front-line chemotherapy with a direct effect on angiogenesis. Additional immunostimulating, immune counter-regulatory and receptor-targeting approaches are also reviewed. The family of epidermal growth factor receptors including epidermal growth factor receptor 1 (HER-1) and 2 (HER-2) are both expressed in ovarian cancer and are the subject of ongoing research and development. The recent disappointing results with 90-yttrium-labeled anti-HMFG by single intraperitoneal administration have left the radiopharmaceutical field without a Phase III candidate. Identification of novel targets may advance this therapeutic area in the future. The rapid advances in the fields of immunoregulation and tumor biology should permit an accelerated introduction of antibodies for the treatment of ovarian cancer. These antibodies could complement novel small molecules that are also in development. PMID- 15757442 TI - Vaccination strategies for the prevention of cervical cancer. AB - Infection with high-risk human papillomaviruses (HPVs) is an essential step in the multistep process leading to cervical cancer. There are approximately 120 different types of HPV identified: of these, 18 are high-risk types associated with cervical cancer, with HPV-16 being the dominant type in most parts of the world. The major capsid protein of papillomavirus, produced in a number of expression systems, self assembles to form virus-like particles. Virus-like particles are the basis of the first generation of HPV vaccines presently being tested in clinical trials. Virus-like particles are highly immunogenic and afford protection from infection both in animal models and in Phase IIb clinical trials. A number of Phase III trials are in progress to determine if the vaccine will protect against cervical disease and, in some cases, genital warts. However, it is predicted that these vaccines will be too expensive for the developing world, where they are desperately needed. Another problem is that they will be type specific. Novel approaches to the production of virus-like particles in plants, second-generation vaccine approaches including viral and bacterial vaccine vectors and DNA vaccines, as well as different routes of immunization, are also reviewed. PMID- 15757443 TI - Therapy of testicular cancer: a surgeon's view. AB - Treatment for metastatic testicular cancer has generally met greatest success when it has involved platinum-based chemotherapy and this is widely used for metastatic disease in most centers. However, surgical techniques should not be excluded. Retroperitoneal lymph node dissection has enabled a high cure rate to be achieved when used in conjunction with chemotherapy in patients with more advanced stage cancers. PMID- 15757444 TI - Update on surgical treatment for endometrial cancer. AB - In the USA, carcinoma of the endometrium consistently ranks as the most common malignancy of the female genital tract. Since the majority of cases present with abnormal clinical symptoms, these patients are typically identified at an early, curable stage when the neoplasm is still confined to the uterus. Surgical removal of the organ involved remains the cornerstone of treatment for this disease, and in light of this, surgery has replaced clinical examination as the staging modality of this malignancy. Surgery also appears to have a role in the management of advanced and recurrent disease. The surgical treatment of this disease is the focus of this review. PMID- 15757445 TI - Recent advances in the treatment of testicular cancer. AB - Testicular cancer is remarkable because it is curable by combination cytotoxic chemotherapy even when widely disseminated. Treatment is defined by widely accepted staging and prognostic factors. Three cycles of bleomycin, etoposide and cisplatin has been defined as the current optimum treatment in good prognosis metastatic disease, curing 90-95% of patients. Outcomes are less impressive for patients in intermediate and poor prognostic categories. A number of different approaches, including introduction of new agents and dose intensification, are being investigated to improve outcomes in these patients. Data developed over the last few years have identified increased risks of second malignancy and cardiovascular disease in long-term survivors. This has led to re-evaluation of strategies to manage Stage I patients. In particular, the use of radiotherapy in Stage I seminoma and the need for adjuvant therapy in Stage I nonseminoma are being re-examined. It is anticipated that advances in imaging and prognostic factors will facilitate this process. PMID- 15757446 TI - Role of chemotherapy in the management of epithelial ovarian cancer. AB - The management of ovarian cancer continues to provide major challenges and debates about optimal treatment. For first-line therapy there remain discussions about optimal chemotherapy for early disease, the use of taxanes as standard for advanced newly diagnosed patients, whether there is a definite role for neoadjuvant chemotherapy and the question of maintenance treatment. For relapsed disease, the management hinges around the distinction between platinum-sensitive and -resistant cancer, and the recent AGO-2.5 and ICON-4 studies suggest that treating with carboplatin and paclitaxel or carboplatin and gemcitabine is recommended. Intraperitoneal chemotherapy remains an enigma with at least three studies showing survival advantage; however, there has been no move to incorporate it into standard management of those patients who achieve complete remission after first-line chemotherapy. Finally, neoadjuvant chemotherapy prior to debulking surgery is the subject of several ongoing clinical trials and may turn out to be one of the most important developments since the concept of interval debulking surgery was established and proven in Europe. PMID- 15757447 TI - Matrix metalloproteinases in cancer: comparison of known and novel aspects of their inhibition as a therapeutic approach. AB - Matrix dissolution is a crucial step during tumor progression that converts a premalignant cell to an overtly malignant one. Main players in this step are the various matrix metalloproteinases (MMPs), which differ in substrate specificity and tissue distribution, and thereby also differ in presence and function during various stages of initial and systemic tumor spread. Accordingly, the inhibition of MMP synthesis and/or activity represents novel potential therapeutic strategies for the treatment of cancer patients. Considerable work has already been carried out on synthetic inhibitors of MMP activity, but with little or even adverse effects in recent clinical studies. The reasons may be inappropriate patient populations in too advanced tumor stages, or inappropriate enzymes as targets for inhibition. Upregulation of endogenous tissue inhibitors of MMP (TIMPs) also provided ambiguous results, since TIMPs possess biologic functions in addition to MMP inhibition, for example, TIMP-2 is a main player in the MMP-2 activation cascade. This may explain, at least in part, the adverse effects of TIMP application/upregulation. Other strategies have been sought in order to overcome these problems. These include the downregulation of MMP transcription by cytokines. However, the effects of cytokines (other than MMP inhibition) may also limit the use of this approach. Finally, empiric evidence for control and modulation of MMP transcription and/or activation by several naturally occurring substances, such as flavonoids, green tea polyphenols and curcumin, represent novel options for the control of MMP activity even in early tumor stages. Additionally, these substances have little or no toxic side effects and good bioavailability, and therefore their continuing analysis provides intriguing insight into tumor pathophysiology and possibly new therapeutic options. PMID- 15757448 TI - The future of adjuvant chemotherapy for resected non-small cell lung cancer. AB - Non-small cell lung cancer is a frequent type of cancer, with approximately 1.2 million cases per year expected worldwide. A total of 20-30% of patients with early stage non-small cell lung cancer are amenable to radical surgery, although only 40-50% of these patients are cured. An improvement in survival has never been demonstrated for postoperative radiotherapy. However, a major step forward is several recent large randomized studies that have demonstrated improved survival with postoperative chemotherapy. This review covers the historic data on adjuvant chemotherapy for non-small cell lung cancer, meta-analyses, modern studies with cisplatin-based or other chemotherapy, implications for current clinical practice and guidelines, some practical recommendations and, finally, the questions for future studies. PMID- 15757450 TI - Would you volunteer to be quarantined and infected with influenza virus? PMID- 15757449 TI - Molecular neuro-oncology and the development of targeted therapeutic strategies for brain tumors. Part 4: p53 signaling pathway. AB - Brain tumors are a diverse group of malignancies that remain refractory to conventional treatment approaches. Molecular neuro-oncology has now begun to clarify the transformed phenotype of brain tumors and identify oncogenic pathways that might be amenable to targeted therapy. Loss of the tumor suppressor gene p53 and its encoded protein are the most common genetic events in human cancer and are a frequent occurrence in brain tumors. p53 functions as a transcription factor and is responsible for the transactivation and repression of key genes involved in cell growth, apoptosis and the cell cycle. Mutation of the p53 gene or dysfunction of its signaling pathway are early events in the transformation process of astrocytic gliomas. The majority of mutations are missense and occur in the conserved regions of the gene, within exons 5 through 8. Molecular therapeutic strategies to normalize p53 signaling in cells with mutant p53 include pharmacologic rescue of mutant protein, gene therapy approaches, small molecule agonists of downstream inhibitory genes, antisense approaches and oncolytic viruses. Other strategies include activation of normal p53 activity, inhibition of mdm2-mediated degradation of p53 and blockade of p53 nuclear export. Further development of targeted therapies designed to restore or enhance p53 function, and evaluation of these new agents in clinical trials, will be needed to improve survival and quality of life for patients with brain tumors. PMID- 15757451 TI - HIV susceptibility affected by genes. PMID- 15757454 TI - Tipranavir: a novel second-generation nonpeptidic protease inhibitor. AB - Tipranavir is a new nonpeptidic protease inhibitor and belongs to the class of 4 hydroxy-5, 6-dihydro-2-pyrones. Chemically, tipranavir is based on coumarin and sulfonamide compounds, amongst others. It exhibits potent and specific activity against both HIV-1 and -2. Tipranavir 500 mg in combination with ritonavir 200 mg twice daily results in optimum viral load reduction and suppresses both wild-type and protease inhibitor-resistant virus. It is metabolized by the cytochrome P4503A4 enzyme and its pharmacokinetic parameters are enhanced when combined with ritonavir. Tipranavir is excreted primarily in the feces, with minimal excretion in urine. In early trials, tipranavir/ritonavir was demonstrated to be safe and well tolerated, with mild gastrointestinal side effects. Preliminary data indicate pharmacokinetic interaction with nucleotide reverse transcriptase inhibitors; however, no dose adjustments are recommended at this time. Virologic response is not adequate when combined with other ritonavir-boosted protease inhibitors, and is currently not recommended. As with other protease inhibitors, tipranavir interacts with fluconazole, atorvastatin, clarithromycin and rifabutin and absorption is reduced when taken with antacids and didanosine (enteric coated formulation). Phase III trials are underway to compare the efficacy of tipranavir/ritonavir with other antiretroviral agents. PMID- 15757455 TI - Ertapenem: review of a new carbapenem. AB - The carbapenems are beta-lactam-type antibiotics with an exceptionally broad spectrum of activity. Ertapenem is a new carbapenem developed to address the pharmacokinetic shortcomings (short half-life) of imipenem and meropenem. Ertapenem shares similar structural features with meropenem, including its stability to dehydropeptidase-1, allowing it to be administered without a dehydropeptidase-1 inhibitor. Ertapenem, like imipenem and meropenem, demonstrates broad-spectrum antimicrobial activity against many Gram-positive and -negative aerobes and anaerobes and is resistant to nearly all beta-lactamases, including extended-spectrum beta-lactamases and AmpCs. However, it differs from both imipenem and meropenem in demonstrating limited activity against Enterococcusspp., Pseudomonasaeruginosa and other nonfermentative Gram-negative bacteria commonly associated with nosocomial infections. The extensive protein binding of ertapenem extends the half-life and allows for once-daily dosing. Prospective, multicenter, randomized, double-blind, comparative clinical studies demonstrate similar clinical efficacy of ertapenem compared with other agents. Clinical trials of complicated intra-abdominal infection, acute pelvic infection, complicated skin and soft-structure infection, community-acquired pneumonia and complicated urinary tract infections demonstrated that ertapenem has equivalent efficacy and safety compared with ceftriaxone and piperacillin/tazobactam. Ertapenem is a promising new carbapenem with excellent efficacy and safety for the treatment of a variety of community-acquired infections. It also appears to be of great value as an outpatient parenteral antimicrobial therapy. PMID- 15757457 TI - Interrupting highly active antiretroviral therapy in patients with HIV. AB - Scheduled treatment interruptions are preplanned interruptions of antiretroviral treatment, which may be directed by time (e.g., cycles of 8 weeks on treatment and 8 weeks off treatment); the concentration of CD4+ lymphocytes (the CD4 count); HIV-1 RNA concentration (viral load); or other factors. This review covers the rationale of scheduled treatment interruptions and the different strategies that have been explored. It examines the issue of autovaccination, resistance and other risks and benefits. Scheduled-treatment-interruption studies in three populations are discussed: patients who initiated highly active antiretroviral therapy during acute HIV infection; patients with successfully treated chronic HIV infection; and patients with highly active antiretroviral therapy failure. PMID- 15757456 TI - Nuclear translocation as a novel target for anti-HIV drugs. AB - During recent years, remarkable progress has been achieved in the treatment of patients infected with HIV. This progress involves not only the improvement of previously known drugs but also the introduction of new classes of anti-HIV agents. Currently, drugs targeting virus entry, reverse transcription, integration and maturation are either in clinical use or in the late stages of clinical development. Nonetheless, the high mutation rate of the virus and toxicity of the drugs, which become problematic during prolonged treatment regimens characteristic of anti-HIV therapy, drive the necessity to produce new drugs that will allow physicians to keep the virus at bay in patients on lifelong anti-HIV therapy. Ideally, such drugs would target a new step in the HIV life cycle, thus avoiding crossresistance with older compounds. One such new target for anti-HIV therapy is nuclear translocation--a process critical for HIV replication. In this article, the authors will review recent literature on the mechanisms of HIV nuclear import and will describe compounds that inhibit this step of HIV replication. PMID- 15757458 TI - The ubiquitin-proteasome system in HIV replication: potential targets for antiretroviral therapy. AB - Since the discovery of HIV approximately 20 years ago, more than 60 million individuals have been infected, and AIDS still remains one of the most devastating diseases humankind has ever faced. Unfortunately, there is little hope that an effective vaccine will be developed in the near future. Current antiretroviral treatment is based on drugs that either target the viral enzymes (protease and reverse transcriptase) or the attachment and entry of the virus. Although the introduction of highly active antiretroviral therapy in the mid 1990s has led to a profound reduction in HIV-related morbidity and mortality, the complete eradication of the virus from infected individuals has never been achieved. In addition, these antiviral drugs can induce serious adverse effects, particularly when administered in combination over prolonged treatment periods. A further drawback to these treatments is that with the high mutation rate of HIV, drug-resistant mutants are evolving, particularly when antiretroviral treatment only suppresses virus replication to marginal levels in latently infected cells making up the virus reservoirs in vivo. Cellular genes have much lower mutation rates, and drug-mediated modulation of specific cellular pathways represents an attractive antiviral strategy. Recent findings showing that proteasome inhibitors interfere with budding, maturation and infectivity of HIV have triggered intensive investigation of the hitherto unappreciated function of the ubiquitin proteasome system in HIV replication. It was also observed that, like several other retroviruses, HIV-1 virions contain a small amount of mono-ubiquitinylated Gag proteins. Currently, two E3-type ubiquitin ligases, in addition to one E3 like protein, have been identified as regulators of HIV budding. These ligases might represent interesting targets for therapeutic intervention. PMID- 15757460 TI - Anti-inflammatory activity of ansamycins. AB - Inflammation represents a complex biologic and biochemical process involving cells of the immune system and a plethora of biologic mediators in response to mechanical, chemical or infectious injuries. When mobilization of effector cells and molecules becomes excessive, the beneficial aspect of this response--to limit damage and promote healing, can be overriden, resulting in host-cell and tissue dysfunction. Based on the hypothesis that chronic infections underly some inflammatory diseases, antibacterial therapy has long been assessed in various inflammatory settings. Recently, the anti-inflammatory activity of some antibacterial agents has also been suspected. Of these duel-action drugs, ansamycins represent an interesting family. Although their therapeutic use is restricted to potentially infectious inflammatory diseases, many experimental data suggest that these drugs also possess direct inhibitory activity on some crucial proinflammatory effectors. To date, the potent antimycobacterial activity of the therapeutically useful ansamycins precludes their widespread use in inflammatory diseases. However, biosynthetic manipulation remains an attractive route for the generation of pharmacologically useful analogs. PMID- 15757459 TI - Therapies for HIV and viral hepatitis coinfection. AB - The natural history of chronic viral hepatitis is altered by HIV coinfection. Liver fibrosis rates and clinical features of liver disease develop more rapidly. Although HIV-hepatitis C virus coinfected subjects may progress more rapidly to AIDS, this is probably explained by comorbid illness, substance abuse and socioeconomic circumstances. Safe and virologically active treatment of HIV hepatitis B virus coinfection can be concurrently achieved by the use of highly active antiretroviral therapy regimens containing lamivudine and/or tenofovir. In most cases, highly active antiretroviral therapy represents the most beneficial initial pharmaceutical intervention for HIV-hepatitisC virus coinfection. HepatitisC virus antiviral therapy should, in most cases, be reserved for those achieving HIV RNA suppression and immune restoration from highly active antiretroviral therapy or with nadir CD4 T-lymphocytes above 350 cells/microl. PMID- 15757461 TI - Varicella-zoster vaccine in the USA: success for control of disease severity, but what next? AB - In the period from 1990 to 1994, before the introduction of a varicella vaccine to the USA, approximately 100 deaths in otherwise healthy individuals, children and adolescents under 20 years of age, were attributable to varicella complications. The administration of a single-dose vaccine has now been widespread in the USA for nearly 10 years; however, since the effectiveness of a single dose in children under 13 years of age in an outbreak situation is approximately 80%, consideration of a second booster dose is in progress although not yet recommended. Licensure of a measles-mumps-rubella-varicella vaccine may hasten the recommendation. PMID- 15757462 TI - Otitis media with effusion: an effort to understand and clarify the uncertainties. AB - Otitis media with effusion--defined as the accumulation of middle-ear effusion behind an intact tympanic membrane without signs or symptoms of acute infection- is one of the most common causes of hearing loss in children in developed countries, potentially leading to language deficits. Although treatment of chronic or relapsing otitis media with effusion is considered imperative, none of the preventative or nonsurgical management measures currently available have proven effective. Tympanostomy tube placement remains the recommended treatment option for high-risk children or for cases of unresponsive otitis media with effusion. This can be attributed to the uncertainties surrounding its pathogenesis. Multiple factors and several possible pathogenetic models have been proposed to explain the production and persistence of middle-ear effusion; only a few of them are supported by sufficient evidence. In this review, the authors will present current knowledge on the pathogenesis, consequences, diagnosis and management of otitis media with effusion. An effort will be made to clarify those aspects sufficiently supported by evidence-based studies, and to underline those that remain unfounded. PMID- 15757463 TI - Ocular bacterial infections: current and future treatment options. AB - In this article, common ocular bacterial infections are reviewed, examining bacterial pathogens, antibiotics and antibacterial resistant trends in light of current and future treatment options. Ophthalmologists are fortunate to be able to choose between an array of old and new antibiotics in order to treat bacterial ocular infections. Ophthalmic infections are primarily treated with topical antibiotics applied directly to the eye. Since there are no in vitro susceptibility standards for interpreting ocular bacterial susceptibility, systemic standards are used. There is no immediate need for new ophthalmic antibiotics, but increasing resistance is being seen with the widely used fluoroquinolone antibiotics. PMID- 15757464 TI - Clinical trials: consent in children. PMID- 15757465 TI - Bacteriophages as tools for vaccine and drug development. PMID- 15757468 TI - Adjuvants enhancing an integral immune response to antigens. 15-17 September, 2004, Modern Vaccine/Adjuvant Formulation: impact on future development (MVAF 2004), Prague. PMID- 15757469 TI - The economics of vaccination from a global perspective: present and future. 2-3 December, 2004, Vaccines: all things considered, San Francisco, CA, USA. PMID- 15757470 TI - What can be done to make vaccines more trendy? AB - Incredibly, vaccines currently capture only about 2% of the world's pharmaceutical market, despite the fact that they can be credited with saving more lives and preventing more suffering and disability than any other form of medical activity, with the possible exception of the provision of clean water. PMID- 15757471 TI - Vaccination considerations in the asplenic patient. AB - Overwhelming postsplenectomy sepsis is a rare but devastating and often lethal disease. Although vaccines are available, their proper use may be questioned. Standardization of protocols for the immunization of asplenic patients should be universal, thus, likely improving on their use. This article reviews the vaccines to be administered to the asplenic patient. PMID- 15757473 TI - DNA vaccines for poultry: the jump from theory to practice. AB - DNA vaccines could offer a solution to a number of problems faced by the poultry industry; they are relatively easy to manufacture, stable, potentially easy to administer, can overcome neonatal tolerance and the deleterious effects of maternal antibody, and do not cause disease pathology. Combined with this, in ovo vaccination offers the advantage of reduced labor costs, mass administration and the induction of an earlier immune response. Together, this list of advantages is impressive. However, this combined technology is still in its infancy and requires many improvements. The potential of CpG motifs, DNA vaccines and in ovo vaccination, however, can be observed by the increasing number of recent reports investigating their application in challenge experiments. CpG motifs have been demonstrated to be stimulatory both in vitro and in vivo. In addition, DNA vaccines have been successfully delivered via the in ovo route, albeit not yet through the amniotic fluid. Lastly, a recent report has demonstrated that a DNA vaccine against infectious bronchitis virus administered via in ovo vaccination, followed by live virus boost, can slightly improve on the protective effect induced by the live virus alone. Therefore, DNA vaccination via the in ovo route is promising and offers potential as a poultry vaccine, however, efficacy needs to be improved and the costs of production reduced before it is likely to be beneficial to the poultry industry in the long term. PMID- 15757474 TI - Fowlpox virus as a recombinant vaccine vector for use in mammals and poultry. AB - Live vaccines against fowlpox virus, which causes moderate pathology in poultry and is the type species of the Avipoxvirus genus, were developed in the 1920s. Development of recombinant fowlpox virus vector vaccines began in the 1980s, for use not only in poultry, but also in mammals including humans. In common with other avipoxviruses, such as canarypox virus, fowlpox virus enters mammalian cells and expresses proteins, but replicates abortively. The use of fowlpox virus as a safe vehicle for expression of foreign antigens and host immunomodulators, is being evaluated in numerous clinical trials of vaccines against cancer, malaria, tuberculosis and AIDS, notably in heterologous prime-boost regimens. In this article, technical approaches to, and issues surrounding, the use of fowlpox virus as a recombinant vaccine vector in poultry and mammals are reviewed. PMID- 15757475 TI - Use of Marek's disease vaccines: could they be driving the virus to increasing virulence? AB - Marek's disease (MD) is an economically important neoplastic disease of poultry. MD almost devastated the poultry industry in the 1960s but the disease was brought under control after Marek's disease herpesvirus (MDV) was identified and vaccines were developed. This is the first effective use of an antiviral vaccination to prevent a naturally occurring cancer in any species. MDV infection has many effects. Initially causing a cytolytic infection in B-lymphocytes, MDV infects activated T-lymphocytes where it becomes latent. In susceptible chicken genotypes MDV transforms CD4+ lymphocytes, causing visceral lymphomas and/or neural lesions and paralysis. Fully productive infection and shedding of infectious virus only occurs in the feather-follicle epithelium. Vaccination of newly-hatched chicks with live vaccines has been widely used to successfully control MD since the early 1970s. However, vaccinated chickens still become infected and shed MDV. Vaccine breaks have occurred with regularity and there is evidence that the use of MD vaccines could be driving MDV to greater virulence. MD continues to be a threat and a number of strategies have been adopted such as the use of more potent vaccines and vaccination of the embryonic stage to provide earlier protection. Recombinant MD vaccines are useful vectors and are being exploited to carry both viral and host genes to enhance protective immune responses. The future aim must be to develop a sustainable vaccine strategy that does not drive MDV to increased virulence. PMID- 15757476 TI - Vaccines for fish in aquaculture. AB - Vaccination plays an important role in large-scale commercial fish farming and has been a key reason for the success of salmon cultivation. In addition to salmon and trout, commercial vaccines are available for channel catfish, European seabass and seabream, Japanese amberjack and yellowtail, tilapia and Atlantic cod. In general, empirically developed vaccines based on inactivated bacterial pathogens have proven to be very efficacious in fish. Fewer commercially available viral vaccines and no parasite vaccines exist. Substantial efficacy data are available for new fish vaccines and advanced technology has been implemented. However, before such vaccines can be successfully commercialized, several hurdles have to be overcome regarding the production of cheap but effective antigens and adjuvants, while bearing in mind environmental and associated regulatory concerns (e.g., those that limit the use of live vaccines). Pharmaceutical companies have performed a considerable amount of research on fish vaccines, however, limited information is available in scientific publications. In addition, salmonids dominate both the literature and commercial focus, despite their relatively small contribution to the total volume of farmed fish in the world. This review provides an overview of the fish vaccines that are currently commercially available and some viewpoints on how the field is likely to evolve in the near future. PMID- 15757477 TI - Hydatid disease: vaccinology and development of the EG95 recombinant vaccine. AB - Hydatid disease is a zoonotic parasitic disease that is distributed widely around the world and causes substantial human morbidity and mortality, particularly in developing countries. Reduction of human hydatid disease using anthelmintics, together with changes in human lifestyle and animal management practices, have been unsuccessful in some developing countries where the disease still persists. Substantial progress has been made towards developing a practical, recombinant vaccine in sheep, to interrupt the lifecycle of Echinococcus granulosus and to prevent subsequent transmission from dogs to humans. This review focuses on the scientific advances in the development of a recombinant vaccine for hydatid disease and the remaining challenges facing the widespread use of the vaccine for control of hydatid disease in endemic areas. PMID- 15757478 TI - Recombinant DNA technology for producing new rinderpest virus vaccines. AB - With few exceptions, vaccination aims to control rather than eliminate or eradicate disease. The eradication of smallpox in the 1970s led to two other human diseases, polio and measles, being targeted for eradication by the World Health Organization. In general, animal diseases are ignored by the public, however, recent targeting of the rinderpest virus, the agent of cattle plague, has put this virus on the verge of global extinction. For centuries, this virus was responsible for major cattle plagues in Europe, Asia and Africa. The success of the Global Rinderpest Eradication Program is an illustration of the power of vaccines to alter people's lives economically and socially when used in an internationally coordinated way. In this review, the history of the development of rinderpest vaccines and the new research being undertaken to produce marker vaccines, using recombinant DNA technology and reverse genetics, are described. In addition, the valuable contribution that marker vaccines can make in the final stages of the rinderpest eradication program is outlined. PMID- 15757479 TI - Novel strategies for prevention and treatment of influenza. AB - Influenza viruses continue to be a major health challenge due to antigenic variation in envelope proteins and animal reservoirs for the viruses. Of particular concern is an anticipated influenza pandemic in the near future. Vaccination is currently the most effective means of reducing morbidity and mortality during influenza epidemics. In addition, neuraminidase inhibitors have substantially improved antiviral therapy for influenza. However, influenza infection in children and the elderly remain problematic. Furthermore, major innovations in prevention and therapy will be needed to deal with an influenza pandemic. This review assesses available and investigational antivirals and vaccines for influenza, emphasising novel approaches that may improve ability to cope with infection in children and the elderly or during a pandemic. Some adverse sequelae of influenza appear to relate to impairment or pathogenic activation of immune responses. Exciting recent findings in this area, with relevance to influenza treatment, are reviewed. PMID- 15757481 TI - The silence of the genes: epigenetic disturbances in haematopoietic malignancies. AB - Cancer-associated disturbances of regulated DNA methylation include both global hypomethylation and gene-specific (often even cancer-specific) hypermethylation. Both coexist and have become the subject of intense investigation. In haematological neoplasias, distinct sets of genes, including the p15/INK4b cell cycle inhibitor (mostly in myeloid malignancies) as well as p16/INK4a (only very infrequently in myeloid neoplasia), have been well characterised as to incidence of hypermethylation, concurrent gene inactivation and their re-expression following treatment with DNA methylation inhibitors. Several genes frequently methylated in haematological neoplasias have been studied with respect to their prognostic value. With the advance of low-dose schedules of demethylating agents (explored particularly in the elderly patient population) the rationale for reverting the 'hyper-methylator phenotype' has also prompted in vivo studies of gene reactivation following this type of treatment. However, ubiquitous surrogate markers for the efficacy of this type of treatment need to be developed. These may include reactivated haemoglobin F (HbF), as demethylating agents can result in clinically meaningful induction of HbF in patients with haemoglobinopathies. Because 'cancer testis antigens', which provide powerful signals for T cell cytotoxic activity on solid tumour cells, are usually silenced in leukaemia but can be reactivated in vitro and in vivo, they provide a rationale for an immuno modulatory effect of demethylating therapy. PMID- 15757480 TI - The plastid-derived organelle of protozoan human parasites as a target of established and emerging drugs. AB - Human diseases like malaria, toxoplasmosis or cryptosporidiosis are caused by intracellular protozoan parasites of the phylum Apicomplexa and are still a major health problem worldwide. In the case of Plasmodium falciparum, the causative agent of tropical malaria, resistance against previously highly effective drugs is widespread and requires the continued development of new and affordable drugs. Most apicomplexan parasites possess a single plastid-derived organelle called apicoplast, which offers the great opportunity to tailor highly specific inhibitors against vital metabolic pathways resident in this compartment. This is due to the fact that several of these pathways, being of bacterial or algal origin, are absent in the mammalian host. In fact, the targets of several antibiotics already in use for years against some of these diseases can now be traced to the apicoplast and by knowing the molecular entities which are affected by these substances, improved drugs or drug combinations can be envisaged to emerge from this knowledge. Likewise, apicoplast-resident pathways like fatty acid or isoprenoid biosynthesis have already been proven to be the likely targets of the next drug generation. In this review the current knowledge on the different targets and available inhibitors (both established and experimental) will be summarised and an overview of the clinical efficacy of drugs that inhibit functions in the apicoplast and which have been tested in humans so far will be given. PMID- 15757482 TI - Antineovascular therapy, a novel antiangiogenic approach. AB - Angiogenesis is a crucial event in tumour growth, since the growth of tumour cells depends on the supply of essentials such as oxygen and nutrients. Therefore, suppression of angiogenesis is expected to show potent therapeutic effects on various cancers. Additionally, this 'antiangiogenic therapy' is thought not only to eradicate primary tumour cells, but also suppress tumour metastases through disruption of haematogenous metastatic pathways. Tumour dormancy therapy does not aim to disrupt newly formed angiogenic vessels but aims to inhibit further formation of neovessels through inhibiting certain processes of angiogenesis. This raises a question of whether or not these antiangiogenic agents bring complete cure of tumours as complete cut-off of oxygen and nutrients is not expected by the treatment with these agents. This paper will review a novel antiangiogenic therapy, antineovascular therapy (ANET). ANET is categorised in antiangiogenic therapy but is different from tumour dormancy therapy using conventional angiogenic inhibitors: ANET aims to disrupt neovessels rather than to inhibit neovessel formation. ANET is based on the fact that angiogenic endothelial cells are growing cells and would be effectively damaged by cytotoxic agents when the agents are effectively delivered to the neovessels. The complete eradication of angiogenic endothelial cells may cause complete cut-off of essential supplies to the tumour cells and lead to indirect but strong cytotoxicity instead of cytostasis caused by the inhibition of angiogenesis. For the purpose of ANET, an angiogenic vasculature-targeting probe has been developed, by which cytotoxic anticancer agents are actively delivered to the angiogenic endothelial cells by using drug delivery system (DDS) technology. Another way to damage newly formed vessels by cytotoxic agents is achieved by metronomic-dosing chemotherapy. This chemotherapy shifts the target of chemotherapeutic agents from tumour cells to angiogenic endothelial cells by selective dosing schedule. Similarly, the shift of target from tumour cells to angiogenic endothelial cells enhanced therapeutic efficacy of cancer photo dynamic therapy (PDT): in this antiangiogenic PDT, photosensitizers are delivered more to neovessel endothelial cells than to tumour cells. These therapeutic strategies would be clinically applied in the future. PMID- 15757483 TI - Targets for antiageing drugs. AB - The process of ageing denotes a postmaturational deterioration of cells and organisms with the passage of time, an increased vulnerability to challenges and a decreased ability to survive. Causes may be found in oxidative damage and incomplete cell maintenance, and both of these factors may be considered the primary targets of any antiageing therapy. Levels of lipophylic anti-oxidant defences may be the limiting factor of longevity in ad-libitum fed animals. An appropriate administration of lipophylic antioxidants (e.g., polyunsaturated fatty acids, vitamin E) and the stimulation of cytomembrane and organelle renewal, by calorie restriction and pharmacological intensification of autophagy, are currently available interventions that may extend longevity and retard the onset, as well as aid the treatment, of most age-associated diseases. PMID- 15757484 TI - Modulation of apoptosis as a target for liver disease. AB - Apoptosis mediated via extrinsic or intrinsic pathways is essential for maintaining cellular homeostasis in the liver. The extrinsic pathway is triggered from the cell surface by engagement of death receptors as CD95, TRAIL (TNF related apoptosis inducing ligand) and TNF (tumour necrosis factor) or TGF-beta (transforming growth factor beta) receptors. The intrinsic pathway is initiated from the mitochondria and can be influenced by Bcl-2 family members. Both pathways are intertwined and play a physiological role in the liver. Dysregulation of apoptosis pathways contributes to diseases as hepatocellular carcinoma, viral hepatitis, autoimmune hepatitis, ischaemia-reperfusion injury, iron or copper deposition disorders, toxic liver damage and acute liver failure. The apoptosis defects are often central pathogenetic events; hence molecular mechanisms of apoptosis give not only insight into disease mechanisms but also provide potential corresponding therapeutic candidates in liver disease. The focus of this review is the identification of apoptotic signalling components in the liver as therapeutic targets. PMID- 15757485 TI - The vascular endothelium in diabetes: a practical target for drug treatment? AB - Vascular disease remains a major cause of morbidity and mortality in diabetes mellitus, in spite of recent improvements in outcome, some of which may be modulated by improved endothelial function. Therapeutic strategies aimed directly at preventing, or minimising the extent of, these sequelae are required as an adjunct to treatments directed at normalising the metabolic milieu. The microvasculature, and the endothelium in particular, are early contributors to vascular dysfunction, thus raising the question as to how best to specifically target the endothelium. However, the expansive nature of the microvasculature, the varying demands that tissues have in terms of blood flow, and the heterogeneity that exists amongst cell types in different sites raises potential problems as to the practicality of such an approach. Further-more, temporal and genetic factors in the genesis of diabetic microvascular dysfunction may impact on therapeutic strategies. It is suggested that a systematic approach is required to understand the heterogeneity of the microvasculature, with particular emphasis on relating differences in gene and protein expression with functional properties. Such an approach may then provide the necessary information to allow exploitation of endothelial cell heterogeneity for unique targeted interventions, as well as providing the necessary rationale for pharmacological interventions (both prophylactic and corrective) aimed at the endothelium as a whole. PMID- 15757486 TI - Considering Fas ligand as a target for therapy. AB - About a decade ago, the death factor Fas ligand (FasL) was identified as the natural trigger of Fas/CD95-dependent apoptosis and as an inducer of Fas dependent activation-induced cell death. Meanwhile, it is known that this molecule not only contributes to target cell lysis in the immune system but also to the establishment of immune privilege and tumour survival. Because delivering a specific antiproliferative signal to T lymphocytes is of major biomedical interest, the FasL/Fas system has gained much attention over the last few years. However, only recently it became evident that the biology of FasL is more complex than initially anticipated. FasL displays a complex pattern of inducible and constitutive expression associated with a number of different functions as a death factor or a co-stimulatory/accessory molecule in lymphocyte activation. Thus, side effects are likely to occur following systemic administration of, for example, anti-FasL medication, not only because of the constitutive FasL expression on cells within immune privileged tissues and vascular endothelium. In addition, FasL comes in different forms: as a surface molecule, as a protease shed soluble variant or secreted in vesicles. Because increased levels of soluble FasL (sFasL) have been determined in various immunological and non-immunological diseases, it has been suggested that sFasL might serve as a prognostic or diagnostic marker even though the pathophysiological cause for its enhanced production is hardly known in most cases. This review summarises the current facts and ideas about the clinical and pharmacological potential of FasL and sFasL as targets for therapeutic interventions. PMID- 15757487 TI - New targets in the treatment of anorexia nervosa. AB - The pathophysiology of anorexia nervosa (AN) is complex and involves alterations of serotonin, dopamine and histamine neurotransmitters. In addition, receptor activity is disturbed, presumably in response to the neurotransmitter changes. These alterations are reviewed in relation to symptomatology and outcome of AN. Neuropeptide and peripheral orexigenic and satiety peptide research is in its infancy but holds much promise to shed light on the pathophysiological mechanisms involved in this illness. Current drug therapies have not demonstrated the efficacy desired in the treatment of AN. Current therapies are reviewed and new drug targets are explored. Compounds that interact with serotonin, histamine and dopamine receptors may offer unique treatment opportunities. In the future, the manipulation of peptides may add to the therapeutic potential of pharmacotherapy. PMID- 15757488 TI - GABA and glutamate systems as therapeutic targets in depression and mood disorders. AB - Advances made in diverse areas of neuroscience suggest that neurotransmitter systems, additional to the monoaminergic, contribute to the pathophysiology of mood disorders. This ever accruing body of preclinical and clinical research is providing increased recognition of the contribution made by amino acid neurotransmitters to the neurobiology of mood disorders. This review examines evidence supporting the role of GABA and glutamate in these processes and explores the potential to target these systems in the development of novel compounds; the viability of these agents for treatment-related co-morbidities will also be considered. PMID- 15757489 TI - Fc receptor targeting in the treatment of allergy, autoimmune diseases and cancer. AB - Immune activation and inhibitory receptors play an important role in the maintenance of an adequate activation threshold of various cells in our immune system. Analyses of murine models show that the inhibitory Fcreceptor, FcgammaRIIB plays an indispensable role in the suppression of anti-body-mediated allergy and autoimmunity. In contrast, the activating-type Fcreceptors (FcRs) are essential for the development of these diseases, suggesting that regulation of inhibitory or activating FcR is an ideal target as a therapeutic agent. In addition, recent crystal structural analyses of FcR-Ig-Fc fragment complexes provide an effective approach for developing FcR-targeting drugs. This review summarises recent advances of FcR, which were mainly obtained by murine studies, and highlights novel antibodies as possible FcR-targeting therapies for allergy, autoimmune diseases and cancer. PMID- 15757490 TI - NF-kappa B--a target in the inflammation of bone destruction. PMID- 15757491 TI - B cells and autoimmunity 2004: new concepts and therapeutic perspectives. AB - Recently, studies of B-cell physiology have continued to provide new and surprising insights into the nature of autoimmunity, highlighting novel potential immunointervention strategies. The meeting on 'B cells and autoimmunity: new concepts and therapeutic perspectives' brought together basic scientists and clinicians with research interests in a range of autoimmune diseases. Recent advances in different facets of B-cell biology were discussed in the prospect of understanding autoimmunity, and significant advances in our understanding of the mechanisms that regulate the autoimmune response at the B cell-level were described. Even though no single message emerged, it is clear that the B lymphocyte is truly destined to become a therapeutic target for the treatment of autoimmune disease. PMID- 15757492 TI - Home office project licence information goes online: increased transparency about animal experiments or a mere attempt to silence critics? PMID- 15757493 TI - Animals and alternatives: societal expectations and scientific need. AB - As Russell and Burch suggested more than 40 years ago, the most humane science is the best science. The path ahead is clear: pain and distress must be eliminated in animal experiments or reduced to an absolute minimum, and, as scientists, we must use the most humane approaches in our research. To accomplish the best science, we must train those who come after us in the principles and practice of humane science. PMID- 15757494 TI - Growth and characterisation of primary bovine colon epithelial cells in vitro. AB - Epithelial crypts from the bovine colon were obtained by using a combined mechanical and enzymatic isolation method, followed by differential D-sorbitol gradient centrifugation. By using this isolation technique, a pure fraction of epithelial crypts with minimal mesenchymal contamination was obtained. The crypts were seeded in collagen-coated plastic flasks. The attached epithelial cells proliferated and formed a confluent monolayer after 6 days in culture. Under low serum culture conditions (1% fetal calf serum), the cells had a population doubling time of 21-22 hours. During the culture period, the colonocytes were characterised morphologically and enzymatically. The morphology of the cultured cells was confirmed by scanning electron microscopy and transmission electron microscopy. The presence of microvilli, tight junctions and desmosomes demonstrated the ability of the cultured cells to restore an epithelial-like cell monolayer. The epithelial origin of the cells was demonstrated by labelling the cells with antibodies against epithelial-specific cytokeratins 7 and 13. The functional integrity of the cells was evaluated by measuring various marker enzymes (gamma-glutamyltranspeptidase, acid phosphatase, alkaline phosphatase, NADH-dehydrogenase) and membrane-associated Na+-K+-ATPase activity. Membrane integrity was determined by measuring the leakage of lactate dehydrogenase into the culture medium. This new culture system for bovine colon epithelial cells could be used as an in vitro model of the colon epithelium in physiological and toxicological studies. PMID- 15757495 TI - The mandatory CAM testing of cells and scaffolds for tissue engineering: benefits for the three Rs of cooperation with the vaccine industry. AB - In cooperation with BAXTER Vaccine AG, which supplies incubated special pathogen free chicken eggs (including a full veterinary record), a permanent hen's egg chorio-allantoic membrane test (HET-CAM) unit has been established, where angiogenesis testing, cell culture, and digital and histological analyses are performed. At the Core Unit for Biomedical Research, the location of the animal testing facility of the Medical University Vienna, cell-scaffold constructs must be evaluated in vitro and in ovo prior to eventual in vivo tissue engineering experiments. The animal testing advisory committee requires that new test proposals are first evaluated by using cell culture and HET-CAM models. Approvals for in vivo experiments are postponed and not issued prior to in vitro/in ovo evaluation. Examples are presented of protocols planned for in vivo studies on cell seeded scaffolds, which were refined after in vitro/in ovo evaluations. PMID- 15757497 TI - Toward a more humane intracerebral pathogenicity index that does not compromise virus pathogenicity ratings. AB - Determination of the intracerebral pathogenicity index (ICPI) of a virus requires tracking sick animals until death or until the eighth day of the standard observation period. A more humane procedure would be to painlessly kill sick animals, but such intervention in the standard protocol could compromise the rating of the virus. In this paper, a modified scoring system is proposed, whereby humanely killed animals are given a score that, on average, does not alter the ICPI. The variance of an animal's contribution to the optimum modified ICPI is never greater than the variance of its contribution to the standard ICPI. PMID- 15757496 TI - Development of a size-dependent aerosol deposition model utilising human airway epithelial cells for evaluating aerosol drug delivery. AB - Aerosol delivery to the airways of the human respiratory tract, followed by absorption, constitutes an alternative route of administration for compounds unsuitable for delivery by conventional oral and parenteral routes. The target for aerosol drug delivery is the airways epithelium, i.e. tracheal, bronchial, bronchiolar and alveolar cells, which become the site of drug deposition. These epithelial layers also serve as a barrier to the penetration of inhaled material. An in vitro model for aerosol deposition and transport across epithelia in the human airways may be a good predictor of in vivo disposition. The present preliminary studies begin an investigation that blends the dynamics of aerosol delivery and the basis of an in vitro simulated lung model to evaluate the transport properties of a series of molecular weight marker compounds across human-derived bronchiolar epithelial cell monolayers. An Andersen viable cascade impactor was used as a delivery apparatus for the deposition of size-segregated particles onto monolayers of small airway epithelial cells and Calu-3 cells. It was shown that these cell layers can withstand placement in the impactor, and that permeability can be tested subsequent to removal from the impactor. PMID- 15757498 TI - Cytotoxic and apoptotic effects on hepatocytes of secondary metabolites obtained from lichens. AB - There are a large number of species of Antarctic lichens, and several studies describing the secondary metabolites present in these lichens, as well as the advances in understanding the chemistry of these metabolites, have been reported. In addition, some derivatives displaying interesting antibacterial effects have been described. The cytotoxic and apoptotic effects of 15 secondary metabolites (depsides, depsidones and usnic acid) obtained from Continental (Chilean) and Antarctic lichens were evaluated in primary cultures of rat hepatocytes. Intracellular lactate dehydrogenase release, caspase 3 activation and DNA fragmentation were measured. In this study, we have evaluated a set of markers associated with pivotal steps in the execution phase of apoptosis, in order to detect compounds with apoptotic effects on hepatocytes before significant necrosis takes place. Flow cytometric analysis of DNA fragmentation revealed an increase in apoptotic nuclei with sub-diploid DNA content after the exposure of hepatocytes to sub-cytotoxic concentrations of the compounds. Among these, salazinic acid, stictic acid and psoromic acid displayed significant apoptotic activities. PMID- 15757499 TI - The feasibility of replacing animal testing for assessing consumer safety: a suggested future direction. AB - At present, we are unable to use much of the data derived from alternative (non animal) tests for human health risk assessment. This brief Comment outlines why it is plausible that new paradigms could be developed to enable risk assessment to support consumer safety decisions, without the need to generate data in animal tests. The availability of technologies that did not exist 10 years ago makes this new approach possible. The approach is based on the concept that data and information derived from applying existing and new technologies to non-animal models can be interpreted in terms of harm and disease in man. A prerequisite is that similar data and information generated in a clinical setting are available to permit this "translation". The incorporation of this additional translation step should make it possible to use data and information generated in non-animal models as inputs to risk assessment. The new technologies include genomics, transcriptomics, proteomics and metabonomics. Their application to in vitro and human "models" enables large amounts of data to be generated very quickly. The processing, interpretation and translation of these data need to be supported by powerful informatics capabilities and statistical tools. The use of integrated "systems biology" approaches will further support the interpretation by providing better understanding of the underlying biological complexity and mechanisms of toxicity. Clinical medicine is using the opportunities offered by the new "omics" technologies to advance the understanding of disease. The application of these technologies in clinical medicine will generate massive amounts of data that will need processing and interpretation to allow clinicians to better diagnose disease and understand the patients' responses to therapeutic interventions. Support from clinical epidemiology will be essential. If these data and information can be made generally accessible in an ethical and legal way, they should also permit the "translation" of experimental non-animal data, so that they can then be used in risk assessment. PMID- 15757500 TI - INVITOX 2004: 13th Workshop of the European Society of Toxicology In Vitro, organised in cooperation with the Scandinavian Society for Cell Toxicology. AB - INVITOX 2004, the 13th Workshop of the European Society of Toxicology In Vitro, was held on 8-11 September 2004, in Zegrze, Poland, in cooperation with the Scandinavian Society for Cell Toxicology. The workshop was attended by 112 participants from 19 countries. The programme included 11 main sessions and two round table discussions. The lectures of the invited speakers, together with short oral presentations and posters, covered the most important aspects of current research in the field of in vitro toxicology. ECVAM's strategies for the replacement of animal testing for toxic chemicals by alternative test systems were discussed. PMID- 15757501 TI - Up-regulation of phospholipid hydroperoxide glutathione peroxidase in rat casein induced polymorphonuclear neutrophils. AB - Antioxidant enzymes play key roles in the protection of cells from oxidative damage. Little is known, however, about the expression of antioxidants and/or their roles in PMNs (polymorphonuclear leucocytes), which are thought to suffer from oxidative stress in an inflammation site. In the present paper, we report on the regulation of expression of PHGPx (phospholipid hydroperoxide glutathione peroxidase) and cGPx (cytosolic glutathione peroxidase) in rat PMNs in the inflammation site. PHGPx mRNA levels were much lower in casein-induced peritoneal and carrageenan-induced pleural PMNs just after their collection than in peripheral PMNs. cGPx mRNA was also reduced in the casein-induced PMNs, but not in carrageenan-induced PMNs. Both enzymes with decreased levels in the casein induced PMNs were up-regulated during further 24 h cultivation in vitro and in vivo, with elevation of their protein levels and activities, and reduction of intracellular peroxides. Up-regulation of PHGPx mRNA was attenuated by cycloheximide, a protein synthesis inhibitor, and this effect was cancelled by culturing the cells in the conditioned medium of the cultured casein-induced PMNs. This latter effect was attenuated by pre-treatment with anti-GRO (growth regulated oncogene) antibody. Recombinant rat GRO could also induce the up regulation in the presence of cycloheximide, demonstrating that GRO may play an important role in the PHGPx up-regulation of casein-induced PMNs. Production of the lipid mediators leukotriene B4 and 5-HETE (5-hydroxyeicosatetraenoic acid) was decreased in the cultured casein-induced PMNs exhibiting PHGPx up-regulation. The evidence obtained indicates that PHGPx activity in the activated PMNs would be related to the appearance of the intrinsic function of PMNs in the inflammatory site. PMID- 15757502 TI - Activation of protein synthesis in cardiomyocytes by the hypertrophic agent phenylephrine requires the activation of ERK and involves phosphorylation of tuberous sclerosis complex 2 (TSC2). AB - The hypertrophic Gq-protein-coupled receptor agonist PE (phenylephrine) activates protein synthesis. We showed previously that activation of protein synthesis by PE requires MEK [MAPK (mitogen-activated protein kinase)/ERK (extracellular signal-regulated kinase) kinase] and mTOR (mammalian target of rapamycin). However, it remained unclear whether ERK activation was required and which downstream components were involved in activating mTOR and protein synthesis. Using an adenovirus encoding the MKP3 (MAPK phosphatase 3) to inhibit ERK activity, we demonstrate that ERK is essential for the activation of protein synthesis by PE. Activation and phosphorylation of S6K1 (ribosomal protein S6 kinase 1) and phosphorylation of eIF4E (eukaryotic initiation factor 4E)-binding protein (both are mTOR targets) were also inhibited by MKP3, suggesting that ERK is also required for the activation of mTOR signalling. PE stimulation of cardiomyocytes induced the phosphorylation of TSC2 (tuberous sclerosis complex 2), a negative regulator of mTOR activity. TSC2 was phosphorylated only weakly at Thr1462, but phosphorylated at additional sites within the sequence RXRXX(S/T). This differs from the phosphorylation induced by insulin, indicating that MEK/ERK signalling targets distinct sites in TSC2. This phosphorylation may be mediated by p90RSK (90 kDa ribosomal protein S6K), which is activated by ERK, and appears to involve phosphorylation at Ser1798. Activation of protein synthesis by PE is partially insensitive to the mTOR inhibitor rapamycin. Inhibition of the MAPK interacting kinases by CGP57380 decreases the phosphorylation of eIF4E and PE induced protein synthesis. Moreover, CGP57380+rapamycin inhibited protein synthesis to the same extent as blocking ERK activation, suggesting that MAPK interacting kinases and regulation of mTOR each contribute to the activation of protein synthesis by PE in cardiomyocytes. PMID- 15757504 TI - Cardiotoxicity of digitalis glycosides: roles of autonomic pathways, autacoids and ion channels. AB - 1 Cardiac glycosides have been used for centuries as therapeutic agents for the treatment of heart diseases. In patients with heart failure, digoxin and the other glycosides exert their positive inotropic effect by inhibiting Na(+)-K(+) ATPase, thereby increasing intracellular sodium, which, in turn, inhibits the Na(+)/Ca(2+) exchanger and increases intracellular calcium levels. As the therapeutic index of digitalis is narrow, arrhythmias are common problems in clinical practice. The mechanisms and mediators of these arrhythmias, however, are not completely understood. 2 The involvement of the sympathetic and parasympathetic nervous system in digitalis cardiac toxicity is reviewed. 3 Receptors, channels, exchange systems or other cellular components involved in digitalis-induced cardiotoxicity are also reviewed. 4 Possible mediators of digitalis-induced cardiac toxicity are discussed. 5 Management of digitalis toxicity in patients is summarized. 6 The determination of the possible mediators of digitalis-induced cardiac toxicity will enhance our knowledge and lead to the development of new therapeutic strategies to treat these lethal arrhythmias. PMID- 15757503 TI - Immunodetection of the expression of microsomal proteins encoded by the glucose 6 phosphate transporter gene. AB - Glucose 6-phosphate transport has been well characterized in liver microsomes. The transport is required for the functioning of the glucose-6-phosphatase enzyme that is situated in the lumen of the hepatic endoplasmic reticulum. The genetic deficiency of the glucose 6-phosphate transport activity causes a severe metabolic disease termed type 1b glycogen storage disease. The cDNA encoding a liver transporter for glucose 6-phosphate was cloned and was found to be mutated in patients suffering from glycogen storage disease 1b. While related mRNAs have been described in liver and other tissues, the encoded protein(s) has not been immunologically characterized yet. In the present study, we report (using antibodies against three different peptides of the predicted amino acid sequence) that a major protein encoded by the glucose 6-phosphate transporter gene is expressed in the endoplasmic reticulum membranes of rat and human liver. The protein has an apparent molecular mass of approx. 33 kDa using SDS/PAGE, but several lines of evidence indicate that its real molecular mass is 46 kDa, as expected. The glucose 6-phosphate transporter protein was also immunodetected in kidney microsomes, but not in microsomes derived from human fibrocytes, rat spleen and lung, and a variety of cell lines. Moreover, little or no expression of the glucose 6-phosphate transporter protein was found in liver microsomes obtained from three glycogen storage disease 1b patients, even bearing mutations that do not directly interfere with protein translation, which can be explained by a (proteasome-mediated) degradation of the mutated transporter. PMID- 15757505 TI - Peptide YY administration into the posterior hypothalamic nucleus of the rat evokes cardiovascular changes by non-adrenergic, non-cholinergic mechanisms. AB - 1 Microinjection of peptide YY (PYY) (0.23-2.3 nmol) into the posterior hypothalamic nucleus (PHN) of conscious rats evokes a dose-dependent pressor response and a bradycardia. 2 The increase in mean arterial pressure evoked by 2.3 nmol of PYY was not blocked by intravenous pretreatment with: (i) the nicotinic ganglionic receptor antagonist pentolinium (PENT, 10 mg kg(-1)) alone, or in combination with the muscarinic receptor antagonist methylatropine (MeATR, 1 mg kg(-1)); (ii) the alpha(1)-adrenoceptor antagonist prazosin (PRAZ, 0.2 mg kg(-1)); (iii) the V(1)-vasopressin receptor antagonist [d(CH(2))(5)Tyr(Me)]AVP (AVPX, 20 microg kg(-1)); (iv) the combination of AVPX, PENT and MeATR; (v) the combination of PRAZ, AVPX, PENT, MeATR, and the alpha(2)-adrenoceptor antagonist yohimbine (0.3 mg kg(-1)); or (vi) the angiotensin II type 1 receptor antagonist ZD 7155 (1 mg kg(-1)). 3 Adrenal demedullation inhibited the PYY-evoked responses of drug-naive rats, and rats pretreated with the combination of PENT, MeATR and AVPX. 4 Transection of the splanchnic nerve innervating the adrenal medullae attenuated the bradycardia, as did ZD 7155, but not the PYY-evoked pressor response. 5 Systemic pretreatment of rats with the neuropeptide Y(1) receptor antagonist BIBP 3226 (1 mg kg(-1)) blocked the PYY-evoked cardiovascular changes, but not those evoked by microinjection of carbachol (5.5 nmol) into the PHN. 6 These results suggest that the cardiovascular changes evoked from the PHN by PYY requires the presence of the adrenal medullae, which are stimulated by: (i) a hormone to release an NPY-like substance that evokes the pressor response, and (ii) the splanchnic nerve to evoke the release of a substance that results in the bradycardia. PMID- 15757506 TI - The role of tyrosine kinase-mediated pathways in diabetes-induced alterations in responsiveness of rat carotid artery. AB - 1 G-protein-coupled receptor signalling, including transactivation of receptor tyrosine kinases (RTKs), has been implicated in vascular pathology. However, the role of specific RTKs in the development of diabetes-induced cardiovascular complications is not known. 2 We investigated the ability of a chronic administration of genistein, a broad-spectrum inhibitor of tyrosine kinases (TKs), AG1478, a specific inhibitor of epidermal growth factor receptor (EGFR) TK activity, and AG825, a specific inhibitor of Erb2, to modulate the altered vasoreactivity of isolated carotid artery ring segments to common vasoconstrictors and vasodilators in streptozotocin (STZ)-induced diabetes. 3 In diabetic carotid artery, the vasoconstrictor responses induced by noradrenaline (NE), endothelin-1 (ET-1), and angiotensin II (Ang II), were significantly increased whereas vasodilator responses to carbachol and histamine were significantly reduced. Inhibition of TKs, EGFR or Erb2 pathway did not affect the body weight or agonist-induced vasoconstrictor and vasodilator responses in the non-diabetic control animals. However, inhibition of TKs by genistein, EGFR TK by AG1478 or Erb2 by AG825 treatment produced a significant normalization of the altered agonist-induced vasoconstrictor responses without affecting blood glucose levels. Treatment with diadzein, an inactive analogue of genistein, did not affect the vasoconstrictor and vasodilator responses in the diabetic animals. 4 Treatment with genistein, AG1478 or AG825 resulted in a significant improvement in diabetes-induced impairment in endothelium-dependent relaxation to carbachol and histamine. 5 These data suggest that activation of TK-mediated pathways, including EGFR TK signalling and Erb2 pathway, are involved in the development of diabetic vascular dysfunction in the carotid artery. PMID- 15757507 TI - Doxapram inhibits carotid sinus baroreceptors in rabbits. AB - 1 To clarify the effects of doxapram on the baroreflex, we recorded carotid sinus nerve (CSN) activity in isolated and perfused carotid artery bifurcations of rabbits. 2 The CSN activity due to chemoreceptor stimulation was blocked by resection of the nerve branches from the carotid body. After the resection, the CSN activity was correlated to increase of carotid sinus (CS) pressure. 3 Administration of doxapram reduced the CSN activity originating from baroreceptors. The effect of doxapram on baroreceptors was dose dependent and reversible. 4 It is unlikely that doxapram altered CS wall mechanics because CS pressure did not change in the presence of the drug. 5 We conclude that doxapram acts on the cardiovascular system in part by inhibiting the negative feedback loop that originates in CS baroreceptors. PMID- 15757508 TI - JAtlasView: a Java atlas-viewer for browsing biomedical 3D images and atlases. AB - BACKGROUND: Many three-dimensional (3D) images are routinely collected in biomedical research and a number of digital atlases with associated anatomical and other information have been published. A number of tools are available for viewing this data ranging from commercial visualization packages to freely available, typically system architecture dependent, solutions. Here we discuss an atlas viewer implemented to run on any workstation using the architecture neutral Java programming language. RESULTS: We report the development of a freely available Java based viewer for 3D image data, descibe the structure and functionality of the viewer and how automated tools can be developed to manage the Java Native Interface code. The viewer allows arbitrary re-sectioning of the data and interactive browsing through the volume. With appropriately formatted data, for example as provided for the Electronic Atlas of the Developing Human Brain, a 3D surface view and anatomical browsing is available. The interface is developed in Java with Java3D providing the 3D rendering. For efficiency the image data is manipulated using the Woolz image-processing library provided as a dynamically linked module for each machine architecture. CONCLUSION: We conclude that Java provides an appropriate environment for efficient development of these tools and techniques exist to allow computationally efficient image-processing libraries to be integrated relatively easily. PMID- 15757509 TI - Gene fusions and gene duplications: relevance to genomic annotation and functional analysis. AB - BACKGROUND: Escherichia coli a model organism provides information for annotation of other genomes. Our analysis of its genome has shown that proteins encoded by fused genes need special attention. Such composite (multimodular) proteins consist of two or more components (modules) encoding distinct functions. Multimodular proteins have been found to complicate both annotation and generation of sequence similar groups. Previous work overstated the number of multimodular proteins in E. coli. This work corrects the identification of modules by including sequence information from proteins in 50 sequenced microbial genomes. RESULTS: Multimodular E. coli K-12 proteins were identified from sequence similarities between their component modules and non-fused proteins in 50 genomes and from the literature. We found 109 multimodular proteins in E. coli containing either two or three modules. Most modules had standalone sequence relatives in other genomes. The separated modules together with all the single (un-fused) proteins constitute the sum of all unimodular proteins of E. coli. Pairwise sequence relationships among all E. coli unimodular proteins generated 490 sequence similar, paralogous groups. Groups ranged in size from 92 to 2 members and had varying degrees of relatedness among their members. Some E. coli enzyme groups were compared to homologs in other bacterial genomes. CONCLUSION: The deleterious effects of multimodular proteins on annotation and on the formation of groups of paralogs are emphasized. To improve annotation results, all multimodular proteins in an organism should be detected and when known each function should be connected with its location in the sequence of the protein. When transferring functions by sequence similarity, alignment locations must be noted, particularly when alignments cover only part of the sequences, in order to enable transfer of the correct function. Separating multimodular proteins into module units makes it possible to generate protein groups related by both sequence and function, avoiding mixing of unrelated sequences. Organisms differ in sizes of groups of sequence-related proteins. A sample comparison of orthologs to selected E. coli paralogous groups correlates with known physiological and taxonomic relationships between the organisms. PMID- 15757510 TI - Bacterial vaginosis, vaginal flora patterns and vaginal hygiene practices in patients presenting with vaginal discharge syndrome in The Gambia, West Africa. AB - BACKGROUND: Bacterial vaginosis (BV) - a syndrome characterised by a shift in vaginal flora - appears to be particularly common in sub-Saharan Africa, but little is known of the pattern of vaginal flora associated with BV in Africa. We conducted a study aimed at determining the prevalence of BV and patterns of BV associated vaginal micro-flora among women with vaginal discharge syndrome (VDS) in The Gambia, West Africa. METHODS: We enrolled 227 women with VDS from a large genito-urinary medicine clinic in Fajara, The Gambia. BV was diagnosed by the Nugent's score and Amsel's clinical criteria. Vaginal swabs were collected for T vaginalis and vaginal flora microscopy, and for Lactobacillus spp, aerobic organisms, Candida spp and BV-associated bacteria (Gardnerella vaginalis, anaerobic bacteria, and Mycoplasma spp) cultures; and cervical swabs were collected for N gonorrhoeae culture and C trachomatis PCR. Sera were tested for HIV-1 and HIV-2 antibodies. Sexual health history including details on sexual hygiene were obtained by standardised questionnaire. RESULTS: BV prevalence was 47.6% by Nugent's score and 30.8% by Amsel's clinical criteria. Lactobacillus spp were isolated in 37.8% of women, and 70% of the isolates were hydrogen-peroxide (H202)-producing strains. Prevalence of BV-associated bacteria were: G vaginalis 44.4%; Bacteroides 16.7%; Prevotella 15.2%; Peptostretococcus 1.5%; Mobiluncus 0%; other anaerobes 3.1%; and Mycoplasma hominis 21.4%. BV was positively associated with isolation of G vaginalis (odds-ratio [OR] 19.42, 95%CI 7.91 - 47.6) and anaerobes (P = 0.001 [OR] could not be calculated), but not with M hominis. BV was negatively associated with presence of Lactobacillus (OR 0.07, 95%CI 0.03 - 0.15), and H2O2-producing lactobacilli (OR 0.12, 95% CI 0.05 - 0.28). Presence of H2O2-producing lactobacilli was associated with significantly lower prevalence of G vaginalis, anaerobes and C trachomatis. HIV prevalence was 12.8%. Overall, there was no association between BV and HIV, and among micro organisms associated with BV, only Bacteroides spp. and Prevotella spp. were associated with HIV. BV or vaginal flora patterns were not associated with any of the factors relating to sexual hygiene practices (vaginal douching, menstrual hygiene, female genital cutting). CONCLUSION: In this population, BV prevalence was higher than in corresponding populations in industrialised countries, but the pattern of vaginal micro-flora associated with BV was similar. BV or vaginal flora patterns were not associated with HIV nor with any of the vaginal hygiene characteristics. PMID- 15757511 TI - Application of population pharmacokinetics to cladribine. AB - BACKGROUND: The nucleoside analog cladribine is used for the treatment of a variety of indolent B- and T-cell lymphoid malignancies. The primary aim of the study was to evaluate the population distribution of pharmacokinetic parameters in patients undergoing treatment with cladribine and to detect the influence of different covariates on the pharmacokinetic parameters. METHODS: This pharmacokinetic study presents the results of a retrospective population pharmacokinetic analysis based on pooled data from 161 patients, who were given cladribine in different administration routes in various dosing regimens. The plasma concentrations of cladribine were determined by reversed-phase high performance liquid chromatography using a solid phase extraction with a limit of quantitation of 1 nM using 1 mL of plasma. RESULTS: A three compartment structural model best described the disposition of cladribine. Clearance was found to be 39.3 L/hour, with a large interindividual variability. The half-life for the terminal phase was 16 hours. Bioavailability was 100% and 35% for subcutaneous and oral administration, respectively, with low interindividual variability. None of the investigated covariates were found to be correlated with the pharmacokinetic parameters. CONCLUSION: As interindividual variability in apparent clearance after oral administration was not significantly higher compared to that following infusion, cladribine could be administered orally instead of intravenously if compensated for its lower bioavailability. Individualized dosing on basis of body surface area or weight does not represent an improvement in this study as compared to administering a fixed dose to all patients. PMID- 15757512 TI - BioAfrica's HIV-1 proteomics resource: combining protein data with bioinformatics tools. AB - Most Internet online resources for investigating HIV biology contain either bioinformatics tools, protein information or sequence data. The objective of this study was to develop a comprehensive online proteomics resource that integrates bioinformatics with the latest information on HIV-1 protein structure, gene expression, post-transcriptional/post-translational modification, functional activity, and protein-macromolecule interactions. The BioAfrica HIV-1 Proteomics Resource http://bioafrica.mrc.ac.za/proteomics/index.html is a website that contains detailed information about the HIV-1 proteome and protease cleavage sites, as well as data-mining tools that can be used to manipulate and query protein sequence data, a BLAST tool for initiating structural analyses of HIV-1 proteins, and a proteomics tools directory. The Proteome section contains extensive data on each of 19 HIV-1 proteins, including their functional properties, a sample analysis of HIV-1HXB2, structural models and links to other online resources. The HIV-1 Protease Cleavage Sites section provides information on the position, subtype variation and genetic evolution of Gag, Gag-Pol and Nef cleavage sites. The HIV-1 Protein Data-mining Tool includes a set of 27 group M (subtypes A through K) reference sequences that can be used to assess the influence of genetic variation on immunological and functional domains of the protein. The BLAST Structure Tool identifies proteins with similar, experimentally determined topologies, and the Tools Directory provides a categorized list of websites and relevant software programs. This combined database and software repository is designed to facilitate the capture, retrieval and analysis of HIV-1 protein data, and to convert it into clinically useful information relating to the pathogenesis, transmission and therapeutic response of different HIV-1 variants. The HIV-1 Proteomics Resource is readily accessible through the BioAfrica website at: http://bioafrica.mrc.ac.za/proteomics/index.html. PMID- 15757513 TI - The chalcone butein from Rhus verniciflua Stokes inhibits clonogenic growth of human breast cancer cells co-cultured with fibroblasts. AB - BACKGROUND: Butein (3,4,2',4'-tetrahydroxychalone), a plant polyphenol, is a major biologically active component of the stems of Rhus verniciflua Stokes. It has long been used as a food additive in Korea and as an herbal medicine throughout Asia. Recently, butein has been shown to suppress the functions of fibroblasts. Because fibroblasts are believed to play an important role in promoting the growth of breast cancer cells, we investigated the ability of butein to inhibit the clonogenic growth of small numbers of breast cancer cells co-cultured with fibroblasts in vitro. METHODS: We first measured the clonogenic growth of small numbers of the UACC-812 human breast cancer cell line co-cultured on monolayers of serum-activated, human fibroblasts in the presence of butein (2 microg/mL) or various other modulators of fibroblast function (troglitazone-1 microg/mL; GW9662-1 microM; meloxican-1 microM; and 3,4 dehydroproline-10 microg/mL). In a subsequent experiment, we measured the dose-response effect on the clonogenic growth of UACC-812 breast cancer cells by pre-incubating the fibroblasts with varying concentrations of butein (10 microg/ml-1.25 microg/mL). Finally, we measured the clonogenic growth of primary breast cancer cells obtained from 5 clinical specimens with normal fibroblasts and with fibroblasts that had been pre-treated with a fixed dose of butein (2.5 microg/mL). RESULTS: Of the five modulators of fibroblast function that we tested, butein was by far the most potent inhibitor of clonogenic growth of UACC-812 breast cancer cells co cultured with fibroblasts. Pre-treatment of fibroblasts with concentrations of butein as low as 2.5 microg/mL nearly abolished subsequent clonogenic growth of UACC-812 breast cancer cells co-cultured with the fibroblasts. A similar dose of butein had no effect on the clonogenic growth of breast cancer cells cultured in the absence of fibroblasts. Significantly, clonogenic growth of the primary breast cancer cells was also significantly reduced or abolished when the tumor cells were co-cultured with fibroblasts that had been pre-treated with a fixed dose of butein. CONCLUSION: We conclude that fibroblasts pre-treated with non toxic doses of butein (a natural herbal compound) no longer support the clonogenic growth of small numbers of primary breast cancer cells seeded into co cultures. These results suggest that interference with the interaction between fibroblasts and breast cancer cells by the natural herbal compound, butein, should be further investigated as a novel experimental approach for possibly suppressing the growth of micrometastases of breast cancer. PMID- 15757515 TI - Molecular mechanisms in uterine epithelium during trophoblast binding: the role of small GTPase RhoA in human uterine Ishikawa cells. AB - BACKGROUND: Embryo implantation requires that uterine epithelium develops competence to bind trophoblast to its apical (free) poles. This essential element of uterine receptivity seems to depend on a destabilisation of the apico-basal polarity of endometrial epithelium. Accordingly, a reorganisation of the actin cytoskeleton regulated by the small GTPase RhoA plays an important role in human uterine epithelial RL95-2 cells for binding of human trophoblastoid JAR cells. We now obtained new insight into trophoblast binding using human uterine epithelial Ishikawa cells. METHODS: Polarity of Ishikawa cells was investigated by electron microscopy, apical adhesiveness was tested by adhesion assay. Analyses of subcellular distribution of filamentous actin (F-actin) and RhoA in apical and basal cell poles were performed by confocal laser scanning microscopy (CLSM) with and without binding of JAR spheroids as well as with and without inhibition of small Rho GTPases by Clostridium difficile toxin A (toxin A). In the latter case, subcellular distribution of RhoA was additionally investigated by Western blotting. RESULTS: Ishikawa cells express apical adhesiveness for JAR spheroids and moderate apico-basal polarity. Without contact to JAR spheroids, significantly higher signalling intensities of F-actin and RhoA were found at the basal as compared to the apical poles in Ishikawa cells. RhoA was equally distributed between the membrane fraction and the cytosol fraction. Levels of F actin and RhoA signals became equalised in the apical and basal regions upon contact to JAR spheroids. After inhibition of Rho GTPases, Ishikawa cells remained adhesive for JAR spheroids, the gradient of fluorescence signals of F actin and RhoA was maintained while the amount of RhoA was reduced in the cytosolic fraction with a comparable increase in the membrane fraction. CONCLUSION: Ishikawa cells respond to JAR contact as well as to treatment with toxin A with rearrangement of F-actin and small GTPase RhoA but seem to be able to modify signalling pathways in a way not elucidated so far in endometrial cells. This ability may be linked to the degree of polar organisation observed in Ishikawa cells indicating an essential role of cell phenotype modification in apical adhesiveness of uterine epithelium for trophoblast in vivo. PMID- 15757517 TI - A Taxonomic Search Engine: federating taxonomic databases using web services. AB - BACKGROUND: The taxonomic name of an organism is a key link between different databases that store information on that organism. However, in the absence of a single, comprehensive database of organism names, individual databases lack an easy means of checking the correctness of a name. Furthermore, the same organism may have more than one name, and the same name may apply to more than one organism. RESULTS: The Taxonomic Search Engine (TSE) is a web application written in PHP that queries multiple taxonomic databases (ITIS, Index Fungorum, IPNI, NCBI, and uBIO) and summarises the results in a consistent format. It supports "drill-down" queries to retrieve a specific record. The TSE can optionally suggest alternative spellings the user can try. It also acts as a Life Science Identifier (LSID) authority for the source taxonomic databases, providing globally unique identifiers (and associated metadata) for each name. CONCLUSION: The Taxonomic Search Engine is available at http://darwin.zoology.gla.ac.uk/~rpage/portal/ and provides a simple demonstration of the potential of the federated approach to providing access to taxonomic names. PMID- 15757514 TI - Bioinformatics analysis of the locus for enterocyte effacement provides novel insights into type-III secretion. AB - BACKGROUND: Like many other pathogens, enterohaemorrhagic and enteropathogenic strains of Escherichia coli employ a type-III secretion system to translocate bacterial effector proteins into host cells, where they then disrupt a range of cellular functions. This system is encoded by the locus for enterocyte effacement. Many of the genes within this locus have been assigned names and functions through homology with the better characterised Ysc-Yop system from Yersinia spp. However, the functions and homologies of many LEE genes remain obscure. RESULTS: We have performed a fresh bioinformatics analysis of the LEE. Using PSI-BLAST we have been able to identify several novel homologies between LEE-encoded and Ysc-Yop-associated proteins: Orf2/YscE, Orf5/YscL, rORF8/EscI, SepQ/YscQ, SepL/YopN-TyeA, CesD2/LcrR. In addition, we highlight homology between EspA and flagellin, and report many new homologues of the chaperone CesT. CONCLUSION: We conclude that the vast majority of LEE-encoded proteins do indeed possess homologues and that homology data can be used in combination with experimental data to make fresh functional predictions. PMID- 15757516 TI - Mice have a transcribed L-threonine aldolase/GLY1 gene, but the human GLY1 gene is a non-processed pseudogene. AB - BACKGROUND: There are three pathways of L-threonine catabolism. The enzyme L threonine aldolase (TA) has been shown to catalyse the conversion of L-threonine to yield glycine and acetaldehyde in bacteria, fungi and plants. Low levels of TA enzymatic activity have been found in vertebrates. It has been suggested that any detectable activity is due to serine hydroxymethyltransferase and that mammals lack a genuine threonine aldolase. RESULTS: The 7-exon murine L-threonine aldolase gene (GLY1) is located on chromosome 11, spanning 5.6 kb. The cDNA encodes a 400-residue protein. The protein has 81% similarity with the bacterium Thermotoga maritima TA. Almost all known functional residues are conserved between the two proteins including Lys242 that forms a Schiff-base with the cofactor, pyridoxal-5'-phosphate. The human TA gene is located at 17q25. It contains two single nucleotide deletions, in exons 4 and 7, which cause frame shifts and a premature in-frame stop codon towards the carboxy-terminal. Expression of human TA mRNA was undetectable by RT-PCR. In mice, TA mRNA was found at low levels in a range of adult tissues, being highest in prostate, heart and liver. In contrast, serine/threonine dehydratase, another enzyme that catabolises L-threonine, is expressed very highly only in the liver. Serine dehydratase-like 1, also was most abundant in the liver. In whole mouse embryos TA mRNA expression was low prior to E-15 increasing more than four-fold by E-17. CONCLUSION: Mice, the western-clawed frog and the zebrafish have transcribed threonine aldolase/GLY1 genes, but the human homolog is a non-transcribed pseudogene. Serine dehydratase-like 1 is a putative L-threonine catabolising enzyme. PMID- 15757518 TI - Complex pediatric elbow injury: an uncommon case. AB - BACKGROUND: There is paucity of literature describing complex elbow trauma in the pediatric population. We described a case of an uncommon pediatric elbow injury comprised of lateral condyle fracture associated with posterolateral dislocation of elbow. CASE PRESENTATION: A 12-year-old boy sustained a direct elbow trauma and presented with Milch type II lateral condyle fracture associated with posterolateral dislocation of elbow. Elbow dislocation was managed by closed reduction. The elbow stability was assessed under general anaesthesia, followed by open K-wiring for the lateral condylar fracture fixation. The patient had an uneventful recovery with an excellent outcome at 39 months follow-up. CONCLUSION: Complex pediatric elbow injuries are quite unusual to encounter, the management of such fractures can be technically demanding. Concomitant elbow dislocation should be managed by closed reduction followed by open reduction and internal fixation (K-wires or cannulated screws) of the lateral condyle fracture. PMID- 15757519 TI - Evaluation of SLC11A1 as an inflammatory bowel disease candidate gene. AB - BACKGROUND: Significant evidence suggests that a promoter polymorphism within the gene SLC11A1 is involved in susceptibility to both autoimmune and infectious disorders. The aim of this study was to evaluate whether SLC11A1 has a role in the susceptibility to inflammatory bowel disease (IBD) by characterizing a promoter polymorphism within the gene and two short tandem repeat (STR) markers in genetic proximity to SLC11A1. METHODS: The studied population consisted of 484 Caucasians with IBD, 144 population controls, and 348 non-IBD-affected first degree relatives of IBD patients. IBD subjects were re-categorized at the sub disease phenotypic level to characterize possible SLC11A1 genotype-phenotype correlations. Polymorphic markers were amplified from germline DNA and typed using gel electrophoresis. Genotype-phenotype correlations were defined using case-control, haplotype, and family-based association studies. RESULTS: This study did not provide compelling evidence for SLC11A1 disease association; most significantly, there was no apparent evidence of SLC11A1 promoter allele association in the studied Crohn's disease population. CONCLUSION: Our results therefore refute previous studies that have shown SLC11A1 promoter polymorphisms are involved in susceptibility to this form of IBD. PMID- 15757520 TI - A qualitative study of the perceptions and experiences of Pre-Registration House Officers on teamwork and support. AB - BACKGROUND: Following the implementation of a new final Year 5 curriculum in one medical school we carried out a study to explore the experience of the transition from final student year to Pre-Registration House Officer (PRHO). This study looks at the experiences of two successive cohorts of PRHOs in relation to team work, support and shared responsibility in their transition from final year students to qualified doctors. The involvement of PRHOs in teams is likely to change in the development of Foundation programmes. METHODS: A qualitative study with semi-structured interviews with 33 PRHOs, stratified by gender, ethnicity and maturity, from two study cohorts, qualifying in 2001 and 2002, from one medical school in the UK, in their first three months following medical graduation. RESULTS: Most PRHOs reported positive experiences for their inclusion as a full member of their first ward teams. This contributed to their increasing confidence and competence in this early period of career transition. However, a number of organisational barriers were identified, e.g. incomplete teams, shift work, which produced problems in their integration for one third of newly qualified doctors. CONCLUSION: Recently introduced policies, intended to improve the working lives of newly qualified doctors have produced both benefits and unintended adverse impacts on PRHOs. The changes of the new PRHO Foundation programme will have further impact. Foundation doctors may need to relate to wider teams with more interaction and less protection. Such changes will need to be managed carefully to protect the PRHO at a vulnerable time. PMID- 15757523 TI - [Inhibitory effect of hTERT dsRNA on telomerase activity in lung carcinoma cell line A549]. AB - BACKGROUND & OBJECTIVE: RNA interference (RNAi) is a new technology in gene study. The mechanism of RNAi is that double-stranded RNA (dsRNA) can band target mRNA and decompose it. This study was to assess possibility and specificity of dsRNA on suppressing human telomerase reverse transcriptase (hTERT) in lung carcinoma cells, investigate its effect on cell proliferation to confirm whether it has unspecific killing activity on mammalian cells, and explore its application in lung cancer research and treatment. METHODS: Sequences of 2 exons and 1 intron of hTERT gene were amplified by reverse transcription-polymerase chain reaction (RT-PCR) or PCR. The sense and antisense cDNA sequences were connected in a tandem manner, and the whole fragment was inserted into pCI-neo mammalian expression vector to construct the dsRNA expression vector, and then transfected into lung carcinoma cell line A549. The expression of hTERT was detected by RT-PCR and Western blot. Telomerase activity was measured by telomerase repeat amplification protocol (TRAP). Cell morphology was observed, and cell proliferation was assessed under invert microscope. RESULTS: After transfection of 2 exon fragments of hTERT dsRNA, mRNA and protein expression of hTERT and telomerase activity in A549 cells were suppressed, cell proliferation was markedly inhibited. Meanwhile, dsRNA didn't show unspecific toxic activity on A549 cells. CONCLUSIONS: hTERT dsRNA can specifically silent hTERT gene, inhibit telomerase activity and proliferation of A549 cells. hTERT dsRNA might be a potential method of gene therapy for lung cancer. PMID- 15757521 TI - A configuration space of homologous proteins conserving mutual information and allowing a phylogeny inference based on pair-wise Z-score probabilities. AB - BACKGROUND: Popular methods to reconstruct molecular phylogenies are based on multiple sequence alignments, in which addition or removal of data may change the resulting tree topology. We have sought a representation of homologous proteins that would conserve the information of pair-wise sequence alignments, respect probabilistic properties of Z-scores (Monte Carlo methods applied to pair-wise comparisons) and be the basis for a novel method of consistent and stable phylogenetic reconstruction. RESULTS: We have built up a spatial representation of protein sequences using concepts from particle physics (configuration space) and respecting a frame of constraints deduced from pair-wise alignment score properties in information theory. The obtained configuration space of homologous proteins (CSHP) allows the representation of real and shuffled sequences, and thereupon an expression of the TULIP theorem for Z-score probabilities. Based on the CSHP, we propose a phylogeny reconstruction using Z-scores. Deduced trees, called TULIP trees, are consistent with multiple-alignment based trees. Furthermore, the TULIP tree reconstruction method provides a solution for some previously reported incongruent results, such as the apicomplexan enolase phylogeny. CONCLUSION: The CSHP is a unified model that conserves mutual information between proteins in the way physical models conserve energy. Applications include the reconstruction of evolutionary consistent and robust trees, the topology of which is based on a spatial representation that is not reordered after addition or removal of sequences. The CSHP and its assigned phylogenetic topology, provide a powerful and easily updated representation for massive pair-wise genome comparisons based on Z-score computations. PMID- 15757524 TI - [Effects of CD151 on migration of human tongue squamous carcinoma cell line Tca8113]. AB - BACKGROUND & OBJECTIVE: Over-expression of CD151 gene in tumor tissues may be associated with metastasis and poor prognosis of cancer, but the mechanism is unknown. This study was designed to determine the effects of CD151 gene on migration of human tongue squamous carcinoma cell line Tca8113. METHODS: Functional fragment of CD151 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), and inserted into plasmid adeno-associated virus (pAAV) vector in sense direction and antisense direction to construct rAAV CD151 and rAAV-antiCD151, the virus titers were determined by dot blot. Two weeks after transfection, protein level of CD151 in Tca8113 cells was detected by Western blot, the effect of CD151 on migration of Tca8113 cells was detected by Transwell chambers. RESULTS: Titer of rAAV-CD151 was 2x10(11) pfu/ml, titer of rAAV-antiCD151 was 1x10(11) pfu/ml. Transfection of rAAV-CD151 increased expression of CD151 in Tca8113 cells by 108% of control cells, while transfection of rAAV-antiCD151 decreased expression of CD151 in Tca8113 cells by 79% of control cells. Migrated cells count was significantly higher in rAAV-CD151 transfected cells (93.6+/-11.6), and lower in rAAV-antiCD151-transfected cells (24.0+/-4.4) than in untransfected and rAAV-GFP-transfected cells (53.0+/-6.6 and 46.0+/-7.0, P<0.05). CONCLUSIONS: It is an important molecular mechanism of tumor metastasis that over-expression of CD151 promotes migration of tumor cells. rAAV antiCD151, a novel therapeutic tool, can specifically reduce the expression of CD151, and inhibit migration of tumor cells. PMID- 15757525 TI - Knockdown of survivin expression by small interfering RNA induces apoptosis in human breast carcinoma cell line MCF-7. AB - BACKGROUND & OBJECTIVE: survivin, a member of inhibitor of apoptosis protein (IAP) gene family, expresses in various human cancer tissues, and may facilitate tumor cell evasion from apoptosis, and promote aberrant mitotic progression. This study was to investigate cell proliferation and apoptosis status of human breast carcinoma cell line MCF-7 after knockdown of survivin. METHODS: Small interfering RNA was transfected into MCF-7 cells to inhibit expression of survivin. mRNA and protein levels of survivin were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Proliferation inhibition rate of MCF-7 cells was analyzed by MTT assay. Cell apoptosis was assayed by flow cytometry (FCM). RESULTS: The expression of survivin in siRNA-transfected group decreased by 64% in comparison to untransfected group. After treatment of different concentrations of siRNA, proliferation inhibition rate and apoptosis rate of MCF 7 cells were increased. The highest proliferation inhibition rate was 60.9%, and the highest apoptosis rate was 29.0% after treatment of 200 nmol/L of siRNA. CONCLUSION: survivin siRNA might be a useful therapeutic agent for the treatment of breast carcinoma. PMID- 15757526 TI - [Methylation of hMLH1 gene promoter in gastric carcinoma with microsatellite instability]. AB - BACKGROUND & OBJECTIVE: Mismatch repair (MMR) deficiency, characterized by microsatellite instability (MSI), is a major cause of hereditary nonpolyposis colorectal carcinoma (HNPCC). Our previous study showed that loss of MMR protein expression can also be seen in gastric carcinoma, but MMR gene mutation seldom occur. Promoter methylation, a main cause of suppressor gene inactivation, might also lead to defect of MMR gene function. This study was designed to investigate the relationship of hMLH1 gene promoter methylation status to MSI in gastric carcinoma, and analyze its action as a risk factor of carcinogenesis. METHODS: DNA was extracted from 52 samples of gastric carcinoma and their adjacent non cancerous gastric mucosa. Polymerase chain reaction (PCR) was used to amplify microsatellite loci BAT-26, D17S261, D3S1283, D2S123, and D3S1611. MSI was studied by capillary electrophoresis. Methylation of hMLH1 gene promoter was detected by restrictive endonuclease digestion, expression of hMLH1 protein was detected by immunohistochemistry. RESULTS: Of the 52 specimens of gastric carcinoma, 13 with high MSI (MSI-H), 2 with low MSI (MSI-l), and 37 with microsatellite stability (MSS). Frequency of hMLH1 promoter methylation was significantly higher in the 13 specimens of gastric carcinoma with MSI-H than in the 39 specimens of gastric carcinoma with MSI-L or MSS (100% vs. 2.6%, P<0.01). Furthermore, frequency of hMLH1 promoter methylation was significantly higher in the 13 specimens of adjacent non-cancerous gastric mucosa of gastric carcinoma with MSI-H than in the 39 specimens of adjacent non-cancerous gastric mucosa of gastric carcinoma with MSI-L or MSS (46.2% vs. 2.6%, P<0.01). The methylation status was accordant with loss of hMLH1 protein expression. Methylation of hMLH1 gene promoter had no relation with differentiation and clinical stage of gastric carcinoma. CONCLUSION: Methylation of hMLH1 gene promoter exists in gastric carcinoma tissues with MSI-H, and their adjacent non-cancerous gastric mucosa, it may be a risk factor of carcinogenesis of gastric carcinoma. PMID- 15757527 TI - [Molecular mechanism of reversing metastatic phenotype in human high-metastatic large cell lung cancer cell line L9981 by nm23-H1]. AB - BACKGROUND & OBJECTIVE: nm23-H1, a tumor metastasis suppressive gene, can reverse tumor metastasis phenotype. But the molecular mechanism of nm23-H1 in inhibiting or reversing metastasis of lung cancer is unclear. This study was to explore the molecular mechanism of nm23-H1 in reversing metastasis phenotype of lung cancer. METHODS: nm23-H1 gene and pLXSN were separately transfected into human lung cancer cell line L9981. Proliferation of L9981, L9981-pLXSN, and L9981-nm23-H1 cells was detected by MTT assay, cell invasive ability was detected by modified Boyden chamber. Tumorigenesis and experimental lung metastasis were determined in vivo. mRNA and protein levels of beta-catenin, E-Cadherin, CD44S, CD44V6, matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-1 (TIMP-1), and vascular endothelial growth factor (VEGF) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: (1)Cell proliferation, clone formation, and invasive ability were significantly lower in L9981-nm23-H1 cells than in L9981 cells [(19.5+/-2.9)% vs. 100%, 10.3+/ 0.7 vs. 21.7+/-1.3, 31.0+/-3.0 vs. 151.0+/-6.3, P<0.01]. (2) The inhibitory rate of tumorigenesis of nude mice was significantly higher in L8891-nm23-H1 group than in L9981 group (85.6% vs. 0%, P<0.001)u the lung metastatic rate was significantly lower in L9981-nm23-H1 group than in L9981 group (0% vs. 100%, P<0.001). (3)nm23-H1 up-regulated mRNA and protein levels of beta-catenin, E Cadherin, and TIMP-1, and down-regulated levels of MMP-2, CD44V6, and VEGF (P<0.01). (4) nm23-H1 up-regulated mRNA level of CD44s, protein level of CD44s didn't change (P>0.05). CONCLUSION: nm23-H1 gene can reverse malignant and metastatic phenotype of L9981 cells through regulating the expressions of lung cancer metastasis-related genes. PMID- 15757528 TI - [Inducing specific antitumor effects on osteosarcoma by fused vaccine of rat dendritic cells]. AB - BACKGROUND & OBJECTIVE: Dendritic cells (DCs) are the strongest antigen presenting cells (APCs). But immune therapy based on DCs for osteosarcoma has seldom been reported. This study was designed to investigate the immunologic potency and antitumor effects of fused vaccine of rat DCs on osteosarcoma. METHODS: Mononuclear cells in rat bone marrow were propagated in vitro under the condition of recombinant granulocytemacrophage colony-stimulating factor (rGM CSF), recombinant interleukin-4 (rIL-4), and recombinant tumor necrosing factor alpha (rTNF-alpha) to obtain DCs. DCs were purified by monoclonal antibody OX62 and magnetic beads, and confirmed by morphologic observation, phenotype determine, and functional experiments. Osteosarcoma cell line UMR-106 was fused with allogeneic or syngeneic DCs by electrofusion to produce the tumor vaccine. Each kind of tumor vaccine was injected subcutaneously into 10 rats to observe its specific antitumor effects. RESULTS: Survival rate of rats after injection of 1x10(7) UMR-106 cells was 70% in allogeneic tumor vaccine group, and 50% in syngeneic tumor vaccine group. The survival rats stood up to the second challenge of 1x10(7) UMR-106 cells 7 weeks after injection. After immunization of 2x10(6) UMR106-DCs, tumors in 60% of the rats bearing pre-established tumor cells diminished or vanished. CONCLUSION: Fused tumor vaccine of UMR-106 cells and allogeneic DCs might induce specific anti-tumor effects on osteosarcoma. PMID- 15757529 TI - [Construction of antisense c-myc recombinant adenovirus and its anti-tumor effects on osteosarcoma cell lines MG-63 and U2OS]. AB - BACKGROUND & OBJECTIVE: c-myc, an oncogene, plays an important role in regulation of cell proliferation, and has been found to be amplified and overexpressed in osteosarcoma. Moreover, it can promote cell transformation, and induce metastasis. This study was to construct recombinant adenovirus encoding antisense c-myc, and to investigate its effects on osteosarcoma cell lines MG-63 (p53 deficient) and U2OS (with wild type p53). METHODS: Recombinant adenovirus Ad-As-c myc encoding antisense c-myc was constructed by gene reconstruction technique, defined by polymerase chain reaction (PCR), and transfected into human osteosarcoma cell lines MG-63 and U2OS. Western blot, acridine orange staining, reverse transcription-PCR (RT-PCR), and flow cytometry (FCM) were used to detect expression of c-myc, proliferation, apoptosis, and cell cycle of MG-63 and U2OS cells. RESULTS: Ad-As-c-myc encoding antisense c-myc was obtained with the titer of 2x10(9) pfu/ml. Ad-As-c-myc significantly inhibited proliferation of both cell lines, while U2OS with wild type p53 was more susceptible to Ad-As-c-myc. Expression of c-myc mRNA was down-regulated in 2 cell lines 48 h after transfection of Ad-As-c-myc. Acridine orange staining and FCM analysis showed that Ad-As-c-myc induced apoptosis of both cell lines, cell cycle analysis showed obvious G(2)/M phase arrest in MG-63 cells,and G1 phase arrest in U2OS cells after transfection of Ad-As-c-myc. CONCLUSION: Ad-As-c-myc could induce apoptosis through both P53-dependent and P53-independent pathways, and inhibit proliferation of osteosarcoma cells. PMID- 15757530 TI - [Selective inducement effect of bacterial redox protein azurin on apoptosis of human osteosarcoma cell line U2OS]. AB - BACKGROUND & OBJECTIVE: Bacterial redox protein azurin can selectively inhibit proliferation, and induce apoptosis in various human cancer cells. These in vitro findings have been confirmed in vivo in nude mice bearing tumor xenograft. Furthermore, azurin could diminish tumor volume in nude mice bearing tumor xenograft with no evidence of toxicity. The mechanism of its action is unknown. This study was designed to explore the effects of azurin on apoptosis of osteosarcoma cell line U2OS, and its molecular mechanism. METHODS: U2OS cells were cultured with different concentrations of azurin. Cell viability was detected by MTT assay. The 50% inhibitory concentration (IC(50)) of azurin was calculated by Bliss method. Apoptotic morphology and apoptotic body of U2OS cells were observed under fluorescent microscope, and transmission electron microscope. DNA ladder was observed through agarose gel electrophoresis. Cell apoptosis was determined by flow cytometry (FCM). Protein levels of Bax, Bcl-2, and Caspase-3 were quantified by Western blot. RESULTS: Azurin inhibited growth and induced apoptosis of U2OS cells in a dose-dependent manner 48 h after treatment, with the IC(50) of(114.54+/-7.65) mg/L. Moreover, its inhibitory effect was significantly higher on U2OS cells than on MG63 cells or L-02 cells under the same concentrations (P<0.05). When treated with 100 or 200 mg/L of azurin for 24 h, U2OS cells showed typical apoptotic morphology, typical DNA ladder bands were observed. No apoptotic feature was observed in control cells. Sub-G1 peak and apoptotic peak of U2OS cells were observed when exposed to 200 mg/L of azurin for 48 h with the apoptosis index of 35.8%, cell cycle was arrested in G1 phase. Bcl 2 was down-regulated when U2OS cells were treated with azurin for 24 h, while Bax and caspase-3 were up-regulated. CONCLUSIONS: Azurin could selectively induce apoptosis of human osteosarcoma U2OS cells. The induction of apoptosis by azurin may be closely associated with down-regulation of Bcl-2, up-regulation of Bax, and activation of Caspase-3. PMID- 15757531 TI - [Correlation of matrix metalloproteinase-3 polymorphism to genetic susceptibility and lymph node metastasis of non-small cell lung cancer]. AB - BACKGROUND & OBJECTIVE: Matrix metalloproteinases (MMPs) might be involved in invasion and metastasis of tumors by degrading extracellular matrix (ECM) and basement membrane (BM). The 5A or 6A single nucleotide polymorphism (SNP) at the 1 171 bp site of promoter region of MMP-3 may modify the transcription and local expression of MMP-3. This study was to investigate correlation of the MMP-3 SNP with genetic susceptibility and lymph node metastasis of non-small cell lung cancer (NSCLC). METHODS: Genotypes of the MMP-3 SNP of 173 NSCLC patients and 350 healthy controls were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Distributions of the MMP-3 genotypes in both NSCLC patients and healthy controls were accorded with Hardy-Weinberg equilibrium (P>0.05). Frequencies of the 6A/6A, 5A/6A, and 5A/5A genotypes were 65.3%, 30.6%, and 4.1% in NSCLC patients, and 67.7%, 30.0%, and 2.3% in healthy controls, whereas frequencies of the 6A and 5A alleles were 79.3% and 20.7% in NSCLC patients, and 82.7% and 17.3% in healthy controls. The overall genotype and allelotype distributions among NSCLC patients and healthy controls were similar (P>0.05). However, stratified analysis found that frequency of the 5A allele was significantly higher in smoking patients than in healthy smokers (21.0% vs. 12.9%, P=0.03). Therefore, smokers with the 5A/6A or 5A/5A genotype have higher risk of developing NSCLC [age and sex adjusted odds ratio (OR) =2.07, 95% confidence interval (CI) =1.13-3.78]. When stratified by pathologic types, no significant difference in MMP-3 genotype or allelotype distribution between adenocarcinoma patients, squamous carcinoma patients and healthy controls was shown. When further stratified by lymphatic metastasis status, frequencies of the 5A allele and the 5A/5A genotype were significantly higher in NSCLC patients with lymphatic metastasis than in NSCLC patients without lymphatic metastasis (22.8% vs. 11.8%, P=0.02u 8.6% vs. 0%, P=0.02). Thus, NSCLC patients with the 5A/5A genotype have higher risk of lymphatic metastasis than NSCLC patients with the 6A/6A genotype (OR=12.38, 95% CI=0.76-202.13). CONCLUSIONS: The 5A allele of MMP 3 might be associated with the increased susceptibility to NSCLC among smokers. The 5A homozygote might increase the risk of lymphatic metastasis in NSCLC patients. PMID- 15757532 TI - [Down-regulation of caveolin-1 in gastric carcinoma and its clinical biological significance]. AB - BACKGROUND & OBJECTIVE: Caveolin-1, a candidate tumor suppressor gene, aberrantly expressed in many kinds of carcinomas. This research was designed to check the expression of Caveolin-1 in non-cancerous gastric mucosa, intestinal metaplasia, atypical hyperplasia, gastric cancer tissues, and gastric cancer cell lines MGC803 and BGC823, and to analyze its clinical biological significance in stepwise gastric carcinogenesis. METHODS: Frozen gastric tissue array-based immunohistochemistry (IHC) was used to examine the expression of Caveolin-1 in 56 specimens of gastric cancer, 29 specimens of non-cancerous mucosa, 11 specimens of intestinal metaplasia, and 7 specimens of atypical hyperplasia. Correlations of Caveolin-1 expression to clinical stage, lymph node metastasis, Lauren's type, and histological type were analyzed. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein levels of Caveolin-1 in 56 specimens of gastric cancer and relevant adjuvant non-cancerous tissue, and cell lines MGC803 and BGC823. RESULTS: Positive rate of Caveolin-1 was significantly lower in gastric cancer than in non-cancerous mucosa, intestinal metaplasia, and atypical hyperplasia (17.9% vs. 84.8%, 81.8%, and 57.1%, P<0.05). Positive rate of Caveolin-1 was lower in advanced gastric cancers than in gastric cancer of early stage (16.0% vs. 33.3%), but the difference was not significant (P>0.05). Positive rate of Caveolin-1 was significantly lower in diffuse gastric cancer than in gastric cancer of intestinal type (7.0% vs. 26.9%, P<0.05), significantly lower in the cases with lymph node metastases than in the cases without lymph node metastases (9.7% vs. 31.8%, P<0.05). Protein and mRNA levels of Caveolin-1 in gastric cancer and relevant adjuvant non-cancerous tissues have no significant difference, but its expression was low in MGC803 and BGC823 cells. CONCLUSION: Progressive down-regulation of Caveolin-1 in gastric epithelial cells is correlated to gastric carcinogenesis. PMID- 15757533 TI - [Effect of angiogenin on angiogenesis in gastric carcinoma]. AB - BACKGROUND & OBJECTIVE: Angiogenin (ANG), an efficient angiogenesis factor, is up regulated in human colorectal carcinoma, pancreatic carcinoma, and other tumors. But the function of ANG in tumor angiogenesis is still unknown. This study was to explore the effect of ANG on angiogenesis of gastric carcinoma. METHODS: The expressions of ANG, CD34, and Ki-67 in 68 specimens of gastric carcinoma were detected by immunohistochemistry. Microvessel density (MVD) was figured out according to the expression of CD34. The proliferation index of carcinoma cells in gastric carcinoma tissue was figured out according to the expression of Ki-67. RESULTS: Of the 68 specimens of gastric carcinoma, 19 showed strongly positive expression of ANG (group A), 3 showed negative expression of ANG (group B). MVD of group A was significantly higher than that of group B (308.4+/-25.6 vs. 196.0+/-31.3, P<0.01). Proliferation index of group A was significantly higher than that of group B (579.6+/-31.4 vs. 341.3+/-84.0, P<0.01). CONCLUSION: Protein level of ANG positively correlates with MVD of gastric carcinoma tissue, and proliferation index of carcinoma cells. PMID- 15757534 TI - [Expression and mutation of Fas-associated death domain (FADD) gene in non-small cell lung cancer]. AB - BACKGROUND & OBJECTIVE: Abnormal expression of Fas-associated death domain (FADD) protein, an important adapter in cell apoptosis signal conduction, may closely relate with tumorigenesis. This study was to detect expression and mutation of FADD gene in non-small cell lung cancer (NSCLC), evaluate its effect on development of NSCLC, and explore the mechanism. METHODS: Polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) was used to detect FADD gene mutation in 62 specimens of NSCLC tissues and 13 specimens of adjacent non-cancerous lung tissues. Immunohistochemistry was used to detect its protein expression. In situ hybridization (ISH) was used to detect FADD mRNA expression in 30 of the 62 specimens of NSCLC tissues. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick labeling (TUNEL) was used to detect apoptotic cells in NSCLC tissues. RESULTS: Of the 62 specimens of NSCLC tissues, 4 cases of stage N2 showed FADD gene mutation. Positive rate of FADD protein in NSCLC tissues was 80.6% (50/62), its protein level positively correlated with differentiation of NSCLC (rs=0.411, P<0.01). Protein level of FADD in NSCLC tissue was significantly higher than that in non-cancerous tissue (P<0.05). Positive rate of FADD mRNA in NSCLC tissue was 80.0%, its concordant rate with positive rate of FADD protein was 88.6% (P>0.05). Apoptotic cells were observed in all specimens of NSCLC, apoptosis indexes of the 4 cases with FADD gene mutation were lower than the mean level, although they showed positive expression of FADD protein. Protein level of FADD was positively related with cell apoptosis of NSCLC (rs=0.599, P<0.001). CONCLUSIONS: FADD gene mutation exists in NSCLC, its mutation and abnormal expression might play a crucial role in carcinogenesis of NSCLC. Protein level of FADD closely correlates with cell apoptosis of NSCLC. PMID- 15757535 TI - [Clinical analysis of resectable breast cancer: a report of 6 263 cases]. AB - BACKGROUND & OBJECTIVE: In Europe and America, breast cancer commonly occurs in women of middle and old age, the median age of patients is about 57 years old. Modified radical mastectomy has taken the place of traditional radical mastectomy, and is called standard radical mastectomy. Patients with breast cancer of early stage commonly receive breast conservative therapy. TNM stage (especially the lymph node status) affects the prognosis, and adjuvant therapy can improve the survival. In China, just a few clinical researches of large amounts of breast cancer patients have been reported. This study was to analyze clinical feature, surgical patterns, treatment outcome of resectable breast cancer, and explore prognostic factors and the effect of adjuvant therapy, for the sake of improving the levels of diagnosis and treatment. METHODS: Records of 6 263 patients with resectable breast cancer, admitted in our hospital from Jun. 1963 to Jun. 2003, were analyzed retrospectively. RESULTS: Of the 6 263 patients, 98.8% were women. Breast cancer occurred most frequently in patients of 40-49 years old (41.0%), especially in patients of 45-49 years old (25.2%). Breast lump was the main clinical manifestation, and occurred in 96.2% of the patients. The 5 , and 10-year survival rates of all patients were 75.2%, and 40.4%. Of the patients in TNM stages 0-I,II, and III, the 5-year survival rates were 96.8%, 73.7%, and 46.4%, respectively the 10-year survival rates were 78.7%, 64.6%, and 33.5%, respectively. The 5-, and 10-year survival rates were higher in lymph node negative group than in lymph node positive group (80.3% vs. 55.6%, and 59.2% vs. 31.9%, P<0.01). There was no significant difference in survival rates of patients received radical mastectomy and modified radical mastectomy since 1980's (P>0.05). Of the 73 patients received breast conservative therapy, no local recurrence or metastasis occurred, with the maximal follow-up of 17 years. Of the patients in stage T2 -T4, the 5-, and 10-year survival rates were significantly higher in adjuvant chemotherapy group than in non-chemotherapy group (78.2% vs. 60.1%, and 48.9% vs. 30.7%, P<0.01). CONCLUSIONS: According to our data, breast cancer most frequently occurred in patients of 45-49 years old. TNM stage (especially the axillary lymph node status)relates to prognosis of breast cancer. The prognosis was worse in the patients with positive lymph node than in the patients with negative Lymph node. The efficacy of modified radical mastectomy equals to that of radical mastectomy, breast conservative therapy can be applied to patients in early stage. Adjuvant chemotherapy and endocrine therapy can improve the survival of resectable breast cancer patients. PMID- 15757536 TI - [Protein expressions of Fas and FasL in B-cell lymphoma and their significances]. AB - BACKGROUND & OBJECTIVE: Fas and FasL have been proved to be the inductional genes of cell apoptosis. Genesis of many tumors relates with functional disorder and abnormal expressions of Fas and FasL. This study was designed to detect protein expressions of Fas and FasL in B-cell non-Hodgkin's lymphoma (B-NHL) and benign lymphoid tissue, and to provide new markers for diagnosis of lymphoma. METHODS: Immunohistochemistry was used to detect protein expressions of Fas and FasL in 92 specimens of B-NHL, and 20 specimens of benign lymphoid tissue. RESULTS: Fas mostly expressed on membrane. FasL mostly expressed in cytoplasm, and partially expressed in nuclei. Positive rate of Fas in B-NHL was 66.3% (61/92), and that of FasL in B-NHL was 67.4% (62/92). Positive rates of both Fas and FasL in benign lymphoid tissue were 60.0% (12/20). There was no significant difference in expressions of Fas and FasL between B-NHL group and benign group (P>0.05), but positive locations of Fas and FasL in these 2 groups are different. Positive rates of Fas and FasL were higher in diffuse large B-cell lymphoma (DLBL) than in follicular lymphoma (FL), and small cell lymphoma (SLL) (87.2% vs. 64.5%, and 31.8%, P<0.05u 89.7% vs. 67.7%, and 27.3%, P<0.05). Positive rates of Fas and FasL in FL were higher than those in SLL. No correlation was found between Fas/FasL expression and patients' gender, age, and tumor location. CONCLUSIONS: The expressions of Fas and FasL are not useful for distinguishing benign lymphoid tissue from lymphoma tissue, while their locational characteristics are valuable for differential diagnosis. The expressions of Fas and FasL are considered valuable in evaluating the malignant grade of B-NHL. PMID- 15757537 TI - [Estimation of hepatic resection volume in hepatocellular carcinoma by ICG(R15) and its relation with postoperative liver failure]. AB - BACKGROUND & OBJECTIVE: It is important to accurately estimate liver reserve function before hepatectomy for hepatocellular carcinoma (HCC), and to determine proper hepatic resection volume during operation, thus, to avoid postoperative liver failure. Recently, indocyanine green retention rate at 15 min (ICG(R15)) has been considered as a sensitive marker for liver reserve function. However, how to evaluate the maximal hepatic resection volume according to liver reserve function, especially ICG(R15), remains controversial. This study was to evaluate the influence of ICG(R15) and hepatic resection volume on postoperative liver failure. METHODS: Records of 225 HCC patients who underwent hepatectomy were retrospectively reviewed. Preoperative ICG(R15) value, intraoperative hepatic resection volume, and postoperative liver function were analyzed. RESULTS: Of the 225 patients, 113 received resection of no more than 1 segment (group A), 76 received resection of 1-2 segments (group B), 36 received resection of more than 2 segments (group C). When ICG(R15) was below 10%, the incidences of postoperative ascites in groups A, B, and C were 21.2% (14/66), 14.3% (8/56), and 15.4% (4/26), respectively (P>0.05); the incidences of jaundice were 3.0% (2/66), 7.1% (4/56), and 15.4% (4/26), respectively (P>0.05). When ICG(R15) was 10%-20%, the incidences of ascites were 26.8% (11/41), 38.9%(7/18), and 50.0% (5/10), respectively (P>0.05); the incidence of jaundice was lower in group A than in group B, and group C [0 (0/41) vs. 27.8% (5/18), and 20.0% (2/10), P<0.01]; 2 patients, who had 2 or 3 segments resected, died of postoperative encephalopathy. When ICG(R15) was more than 20%, ascites occurred in 2 of 6 patients (33.3%) in group Au both of the 2 cases in group B developed postoperative ascites and jaundice, and 1 died. CONCLUSIONS: When ICG(R15) is below 10%, 2 or more segments can be resected safely when ICG(R15) is 10%-20%, 1 segment can be resected safely, 2 or more segments may be resected with great care when ICG(R15) is more than 20%, the patient's general condition and other liver function tests should be taken into consideration if 1 segment is resected, resection of 2 or more segments will cause high surgical risk. PMID- 15757538 TI - [Predictive value of detection of sentinel lymph nodes on lymphatic metastasis of non-small cell lung cancer]. AB - BACKGROUND & OBJECTIVE: The metastasis status of regional lymph node is an important prognostic factor of non-small cell lung cancer (NSCLC). Sentinel lymph node (SLN) mapping and biopsy is a quick and high efficient technique to intraoperatively detect occult micrometastatic disease, however, its application in NSCLC is immature. This study was designed to investigate the feasibility of detecting SLN in patients with NSCLC during radical surgery, and to evaluate its accuracy of predicting metastasis status of regional lymph node. METHODS: Fifty patients with NSCLC underwent SLN detection. During radical operation, 4 ml of 1% isosulfan blue was injected into the lung tissue around the tumor at 3, 6, 9, and 12 o'clock sites. Location and number of blue dyed SLNs were recorded, and compared with pathologic results to calculate the accuracy and false negative rate of SLN detection. RESULTS: Blue dyed SLNs were seen in 33 patients with a detection rate of 66.0%. SLNs located in N1 lymph node of 24 patients (72.7%), in N2 lymph node of 6 patients (18.2%), in both N1 and N2 lymph nodes of 3 patients (9.1%). Approved by pathology, the accuracy of SLN detection was 87.9% (29/33), the sensibility was 73.3% (11/15), the false negative rate was 26.7% (4/15). CONCLUSION: SLN detection is valuable for predicting hilar and mediastinal lymph nodes metastases in NSCLC. PMID- 15757540 TI - [Clinical value of cervical mediastinoscopy in non-small cell lung cancer of stage I]. AB - BACKGROUND & OBJECTIVE: Whether to apply routine mediastinoscopy to non-small cell lung cancer (NSCLC) of stage I remains controversial. This study was to evaluate application of cervical mediastinoscopy in detecting mediastinal lymph nodes status of NSCLC of stage I. METHODS: The cervical mediastinoscopy was performed in 31 patients with NSCLC of stage I prior to thoracotomy from Oct. 2000 to Feb. 2004. Mediastinoscopy-positive (stage N2) patients received neoadjuvant chemotherapy. RESULTS: Of the 31 patients, 2 had disease of stage N2, and 1 had disease of stage N3 (right scalenus lymph node metastasis); positive rate of mediastinosopy was 10% (3/31), sensitivity, specificity, and accuracy of mediastinoscopy were 75%, 100%, and 97%, respectively. CONCLUSION: Mediastinoscopy might be a routine examination for patients with NSCLC of stage I. PMID- 15757539 TI - [Expressions of 8-OH-dG, k-ras, and p53 genes in pleural effusion cells and their clinical significances]. AB - BACKGROUND & OBJECTIVE: Oxidative DNA damage plays an important role in carcinogens-induced carcinogenesis. 8-hydoxy-2deoxy-guanosine (8-OH-dG), a biomarker of oxidative DNA damage, plays important roles in initiation, progression, and prognosis of lung cancer, and closely relates with mutations of k-ras and p53 genes in carcinogenesis of lung tissue. This study was to detect protein expressions of 8-OH-dG, k-ras, and p53 genes in lung cancer tissues, and to analyze their values in distinguished diagnosis of lung cancer. METHODS: Protein levels of 8-OH-dG, k-ras, and p53 in pleural effusion cells from 53 patients with lung cancer, and 53 patients with other benign lung diseases were detected by immunocytochemistry. RESULTS: Positive rates of 8-OH-dG, k-ras, and p53 protein in cancer group were significantly higher than those in benign disease group [75.5% (40/53) vs. 15.1% (8/53), P < 0.01; 64.2% (34/53) vs. 3.8% (2/53), P < 0.01; and 69.8% (37/53) vs. 18.9% (10/53), P < 0.01; respectively]. Protein levels of 8-OH-dG, k-ras, and p53 protein in cancer group were 1.68+/ 1.21, 1.32+/-1.06, and 1.57+/-1.15,respectively. Rank correlation analysis showed that protein expression of 8-OH-dG positively correlated with those of k-ras (RS=0.643, P < 0.01), and p53 (RS=0.827, P < 0.01)u protein expression of k-ras positively correlated with that of p53 (RS=0.897, P < 0.01). CONCLUSIONS: Protein expressions of 8-OH-dG, k-ras, and p53 are up-regulated in pleural effusion cells of lung cancer, and have mutual relations. They may be used as reference markers in diagnosing and screening for lung cancer. PMID- 15757541 TI - [Postoperative complications in lung cancer patients with moderate pulmonary hypofunction]. AB - BACKGROUND & OBJECTIVE: With the development of surgery treatment for lung cancer, the importance of preoperative pulmonary function assessment evoked more and more attention. This research was set to analyze postoperative complications of lung cancer patients with moderate pulmonary hypofunction, and explore associated factors. METHODS: Statistic t test and Chi(2) test were used to compare postoperative complications of 31 patients with moderate pulmonary hypofunction and 62 patients with normal pulmonary function (control group), logistic regression was performed to find associated factors. RESULTS: In the 31 patients with moderate pulmonary hypofunction, hypoxemia (41.9%), arrhythmia or cardiac dysfunction (25.8%), pulmonary inflammation (25.8%) were common postoperative complications; incidence of severe complications, such as respiratory failure, and cardiac failure, was 9.2%; in-hospital mortality was 3.2%. In control group, the occurrence rates were 16.1%, 8.1%, 9.6%, and 3.2%, accordingly, no in-hospital death occurred. Regression analysis showed that age and resection range associated with the occurrence of postoperative complications. CONCLUSIONS: compared with control group, incidences of common postoperative complications in patients with moderate pulmonary hypofunction are remarkably increased, but severe complications and in-hospital mortality are not significantly high. Patients with older age and larger resection range have a high risk of develop complications. PMID- 15757542 TI - [Magnetic resonance imaging manifestations of radiation injury in brain stem and cervical spinal cord of nasopharyngeal carcinoma patients after radiotherapy]. AB - BACKGROUND & OBJECTIVE: It is very important to diagnose the radiation injury in brain stem and cervical spinal cord of patients with nasopharyngeal carcinoma (NPC) after radiotherapy. Magnetic resonance imaging (MRI) manifestations of radiation encephalopathy have been widely reported, while those of radiation injury in brain stem and cervical spinal cord have been seldom reported. This study was to analyze the MRI characteristics of radiation injury in brain stem and cervical spinal cord of patients with NPC after radiotherapy. METHODS: MRI was performed in 60 NPC patients 6 months to 5 years after radiotherapy. The imaging sequences included T(1)-weighted image (T(1)WI), T2-weighted image (T(2)WI), fluid attenuated inversion recovery (FLAIR). All patients received T(1)WI contrast-enhanced scanning. RESULTS: Of the 60 patients, 6 had lesions in cervical spinal cord, 54 had lesions in brain stem. Of the 54 cases of radiation injury in brain stem, most lesions located in pons (20 cases), basis pons and medulla oblongata (26 cases), others located in mesencephalon (3 cases), medulla oblongata (5 cases). All lesions showed hypo- or iso-intense signal on T(1)WI, and hyper-intense signal on T(2)WI. After contrast-enhanced scanning, 11 cases (18.3%) had no enhancement; 49 (81.7%) markedly enhanced with 21 cases (42.9%) of homogenous patchy enhancement, and 28 cases (57.1%) of hetergenerous ringed and patchy enhancement. CONCLUSION: MRI shows clearly the radiation injury in brain stem and cervical spinal cord of NPC patients after radiotherapy. PMID- 15757543 TI - [Mucoepidermoid carcinoma of nasopharynx: a report of twelve cases]. AB - BACKGROUND & OBJECTIVE: Mucoepidermoid carcinoma of nasopharynx, a kind of primary adenocarcinoma of nasopharynx, is rare, and has seldom been reported. This article was to summarize the pathogenesis, clinical features, and treatment outcomes of this disease according to our experiences. METHODS: From Jan. 1975 to Dec. 2003, 12 patients with pathologically confirmed primary mucoepidermoid carcinoma of nasopharynx were treated in our hospital. We analyzed retrospectively their clinical data with literature review. RESULTS: The 12 patients with mucoepidermoid carcinoma of nasopharynx accounted for only 0.026% of all nasopharyngeal cancer patients diagnosed simultaneously in our hospital. Age of getting disease was 20 to 60 years old with male to female ratio of 2:1. Positive rates of Epstein-Barr virus serological tests (VCA-IgA, EA-IgA, and DNA enzyme ratio) were very low [50.0% (6/12), 11.1% (1/9), and 20.0% (1/5)]. Of the 11 patients with follow-up data, 4 received chemotherapy, 4 received surgery plus radiotherapy, and 3 received radiochemotherapy. The 5-, and 10-year survival rates of the 12 patients were 27.3%, and 9.0%. CONCLUSIONS: Mucoepidermoid carcinoma of nasopharynx is a special type of nasopharyngeal carcinoma with specific pathogenesis features. Surgery-predominant of complete lumpectomy combined treatment strategy is possibly accommodating. PMID- 15757545 TI - [Surgical management for occult cervical metastasis of oral tongue squamous cell carcinoma]. AB - BACKGROUND & OBJECTIVE: In oral tongue squamous cell carcinoma (SCC), the management for clinically negative (cN0) neck remains controversial. This study was to explore rational neck management for cN0 cases. METHODS: Clinical data of 187 patients with oral tongue SCC of cN0, who received surgery alone, were retrospectively reviewed. Rules of occult nodal metastasis, factors that impact cervical node metastasis, and prognostic factors were analyzed. Neck recurrences of different treatment groups were compared. RESULT: Incidence of occult nodal metastasis in all patients was 23.0% (43/187). The most common site of occult nodal metastases was ipsilateral level II. About 83.0% of occult nodal metastases were found in ipsilateral levels I, II, and III. Pathologic grade was an independent influencing factor for occult neck metastases. Occult neck metastasis was an independent prognostic factor for oral tongue SCC. Supraomohyoid neck dissection (SOHND) was an effective therapeutic method for oral tongue SCC of cN0; the neck recurrence rate after SOHND was only 6.7%. CONCLUSIONS: SOHND is the effective and safe treatment for oral tongue SCC. PMID- 15757544 TI - [Clinicopathologic features and treatment outcomes of primary hepatic lymphoma: a report of four cases]. AB - BACKGROUND & OBJECTIVE: Primary hepatic lymphoma (PHL), a rare malignancy, has seldom been reported. This article was to investigate clinicopathologic features and treatment of PHL. METHODS: Data of 4 patients with PHL, treated in Cancer Center, Sun Yat-sen University from Nov. 1994 to May 2004, were retrospectively analyzed. RESULTS: Of the 4 patients, 3 were men, and 1 was woman, with a median age of 53 years old; 3 had single focus, and 1 had multi-foci. All patents were positive for HBV antigen, 3 were misdiagnosed preoperatively, and 1 had no clear diagnosis before operation. Pathologic examination revealed that 3 were B-cell non-Hodgkin's lymphoma, and 1 was Hodgkin's lymphoma. Two patients received resection, and 2 received biopsy; all patients received adjuvant chemotherapy postoperatively. CONCLUSIONS: HBV infection might relate with the development of PHL. PHL should be considered for the patients presented liver mass and B symptomatolgy. Resection followed by adjuvant chemotherapy with CHOP regiment seems to be the best option for PHL. PMID- 15757546 TI - [Detection of RNA interference in nasopharyngeal carcinoma cell lines using reporter genes]. AB - BACKGROUND & OBJECTIVE: RNA interference (RNAi) technique is now widely used in studies of gene function, signal transduction pathway, and gene therapy because it can effectively and specifically inhibit gene expression. This study was designed to synthesize small interfering RNA (siRNA) by in vitro transcription, and construct retrovirus vectors to express small hairpin RNA (shRNA), detect RNAi in nasopharyngeal carcinoma cell lines, and to develop a RNAi technique platform. METHODS: siRNAs targeting green fluorescent protein (GFP) and luciferase (Luc) were synthesized by in vitro transcription, while shRNAs targeting GFP and Luc were constructed from pSUPER.retro. Cervical cancer cell line HeLa, nasopharyngeal carcinoma cell lines CNE1, CNE2, and 5-8F were co transfected with siRNAs or shRNAs and reporter gene pEGFP-N1 or pGL3. The expression of GFP was detected by fluorescent microscopy and Western blot. The activity of luciferase was measured by Luciferase Enzyme Assay System. RESULTS: siRNA duplexes with 3' UU overhangs and shRNA specifically silenced GFP expression, while antisense RNA and siRNA without 3' UU overhangs did not trigger RNA interference of GFP. Quantitative luciferase activity analysis showed that siRNA inhibited Luc expression in HeLa, CNE1, CNE2, and 5-8F cell lines with inhibition rates of 91.43%, 78.01%, 90.30%, and 62.85%, respectively. Similarly, the inhibition rate was 78.22% when shRNA targeting Luc was co-transfected into HeLa cell line. CONCLUSIONS: Both siRNAs and shRNAs can induce RNAi. 3' UU overhangs of siRNA may play a role in RNAi. RNAi can be triggered in both nasopharyngeal carcinoma cell lines and HeLa cell line. PMID- 15757547 TI - [Research advancement of endogenous angiogenesis inhibitors]. AB - Angiogenesis is required for invasive tumor growth and metastasis. Inhibition of angiogenesis is considered to be a promising approach of antitumor therapy. Recently, many endogenous angiogenesis inhibitors have been discovered, some of them are currently in various stages of clinical trials. This review focused on the structure, function, and mechanism of endogenous angiogenesis inhibitors, and their potential in treating tumor. PMID- 15757548 TI - Fly transmission of Campylobacter. AB - An annual increase in Campylobacter infection in England and Wales begins in May and reaches a maximum in early June. This increase occurs in all age groups and is seen in all geographic areas. Examination of risk factors that might explain this seasonal increase identifies flies as a potential source of infection. The observed pattern of infection is hypothesized to reflect an annual epidemic caused by direct or indirect contamination of people by small quantities of infected material carried by flies that have been in contact with feces. The local pattern of human illness appears random, while having a defined geographic and temporal distribution that is a function of the growth kinetics of one or more fly species. The hypothesis provides an explanation for the seasonal distribution of Campylobacter infections seen around the world. PMID- 15757549 TI - Disease risks from foods, England and Wales, 1996-2000. AB - Data from population-based studies and national surveillance systems were collated and analyzed to estimate the impact of disease and risks associated with eating different foods in England and Wales. From 1996 to 2000, an estimated 1,724,315 cases of indigenous foodborne disease per year resulted in 21,997 hospitalizations and 687 deaths. The greatest impact on the healthcare sector arose from foodborne Campylobacter infection (160,788 primary care visits and 15,918 hospitalizations), while salmonellosis caused the most deaths (209). The most important cause of indigenous foodborne disease was contaminated chicken (398,420 cases, risk [cases/million servings] = 111; case-fatality rate [deaths/100,000 cases] = 35, deaths = 141). Red meat (beef, lamb, and pork) contributed heavily to deaths, despite lower levels of risk (287,485 cases, risk = 24, case-fatality rate = 57, deaths = 164). Reducing the impact of indigenous foodborne disease is mainly dependent on controlling the contamination of chicken. PMID- 15757550 TI - Rapid identification of emerging pathogens: coronavirus. AB - We describe a new approach for infectious disease surveillance that facilitates rapid identification of known and emerging pathogens. The process uses broad range polymerase chain reaction (PCR) to amplify nucleic acid targets from large groupings of organisms, electrospray ionization mass spectrometry for accurate mass measurements of PCR products, and base composition signature analysis to identify organisms in a sample. We demonstrate this principle by using 14 isolates of 9 diverse Coronavirus spp., including the severe acute respiratory syndrome-associated coronavirus (SARS-CoV). We show that this method could identify and distinguish between SARS and other known CoV, including the human CoV 229E and OC43, individually and in a mixture of all 3 human viruses. The sensitivity of detection, measured by using titered SARS-CoV spiked into human serum, was approximate, equals1 PFU/mL. This approach, applicable to the surveillance of bacterial, viral, fungal, or protozoal pathogens, is capable of automated analysis of >900 PCR reactions per day. PMID- 15757551 TI - Antimicrobial drug prescribing for pneumonia in ambulatory care. AB - To determine patterns and predictors of antimicrobial drug use for outpatients with community-acquired pneumonia, we examined office visit and pharmacy claims data of 4 large third-party payer organizations from 2000 to 2002. After patients with coexisting conditions were excluded, 4,538 patients were studied. Despite lack of coexisting conditions, fluoroquinolone use was commonly observed and increased significantly (p < 0.001) from 2000 to 2002 (24%-39%), while macrolide use decreased (55%-44%). Increased age correlated with increased fluoroquinolone use: 18-44 years (22%), 45-64 years (33%), and > or =65 years (40%) (p < 0.001). Increased use of fluoroquinolones occurred in healthy young and old patients alike, which suggests a lack of selectivity in reserving fluoroquinolones for higher risk patients. Clear and consistent guidelines are needed to address the role of fluoroquinolones in treatment of outpatient community-acquired pneumonia. PMID- 15757552 TI - Ebola virus antibody prevalence in dogs and human risk. AB - During the 2001-2002 outbreak in Gabon, we observed that several dogs were highly exposed to Ebola virus by eating infected dead animals. To examine whether these animals became infected with Ebola virus, we sampled 439 dogs and screened them by Ebola virus-specific immunoglobulin (Ig) G assay, antigen detection, and viral polymerase chain reaction amplification. Seven (8.9%) of 79 samples from the 2 main towns, 15 (15.2%) of 99 samples from Mekambo, and 40 (25.2%) of 159 samples from villages in the Ebola virus-epidemic area had detectable Ebola virus-IgG, compared to only 2 (2%) of 102 samples from France. Among dogs from villages with both infected animal carcasses and human cases, seroprevalence was 31.8%. A significant positive direct association existed between seroprevalence and the distances to the Ebola virus-epidemic area. This study suggests that dogs can be infected by Ebola virus and that the putative infection is asymptomatic. PMID- 15757553 TI - Probable psittacosis outbreak linked to wild birds. AB - In autumn 2002, an outbreak of probable psittacosis occurred among residents of the Blue Mountains district, Australia. We conducted a case-control study to determine independent risk factors for psittacosis by comparing exposures between hospitalized patients and other residents selected randomly from the telephone directory. Of the 59 case-patients with laboratory results supportive of psittacosis, 48 participated in a case-control study with 310 controls. Independent risk factors were residence in the upper Blue Mountains (odds ratio [OR] 15.2, 95% confidence interval [CI] 5.6-41.7), age of 50-64 years (OR 3.9, 95% CI 1.5-10.5), direct contact with wild birds (OR 7.4, 95% CI 2.5-22), and mowing lawns without a grass catcher (OR 3.2, 95% CI 1.3-8.0). Protective equipment is recommended for residents in areas frequented by free-ranging birds if contact with birds and their droppings is likely when performing outdoor activities such as lawn mowing. PMID- 15757554 TI - Effect of regulation and education on reptile-associated salmonellosis. AB - Reptiles have become increasingly common as domestic pets, and with them reptile associated Salmonella infections in humans. From 1990 to 2000, a total of 339 reptile-associated Salmonella cases were reported in Sweden. In 1996, as part of its efforts to adapt its import regulations to those of the European Union, Sweden no longer required certificates stating that imported animals were free of Salmonella. A subsequent increase was noted in the incidence of reptile associated cases from 0.15/100,000 in the period 1990-1994 to 0.79/100,000 in 1996 and 1997. After a public education campaign was begun through the news media, the incidence dropped to 0.46/100,000. Children were the most affected age group among patients (incidence 1.3/100,000). Salmonella enterica serotype Enteritidis was the most frequent serotype (24% of isolates), followed by S. Typhimurium (9% of isolates). Import restrictions and public information campaigns are effective public health measures against reptile-associated salmonellosis. PMID- 15757555 TI - SARS risk perceptions in healthcare workers, Japan. AB - In coping with severe acute respiratory syndrome (SARS), infection control measures are a key aspect of protecting healthcare workers. We conducted a survey concerning perception of risk and countermeasures for SARS in 7 tertiary hospitals in Japan from July through September 2003, immediately after the SARS epidemic in neighboring countries. Based on 7,282 respondents out of 9,978 questionnaires administered, we found the perception of risk to be relatively high and the perception of countermeasures at the institutional level to be relatively low. Knowledge of preventive measures, concept of (opinions regarding) institutional measures, and perception of risk differed substantially among the 3 job categories, notably between physicians and nurses. The concept of institutional measures was the most important predictor of individual perception of risk. In view of the potential for future epidemics, planning and implementing institutional measures should be given a high priority. PMID- 15757556 TI - Longitudinally profiling neutralizing antibody response to SARS coronavirus with pseudotypes. AB - The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) is a major target for neutralizing antibodies. Retroviral SARS-CoV S pseudotypes have been constructed and used to develop an in vitro microneutralization assay that is both sensitive and specific for SARS-CoV neutralizing antibodies. Neutralization titers measured by this assay are highly correlated to those measured by an assay using replication-competent SARS-CoV. No cross-neutralization occurred with human sera known to contain antibodies to coronavirus strains OC43 and 229E. The pseudotype assay was used to profile neutralizing antibody responses against SARS-CoV S in sequential serum samples taken from 41 confirmed SARS patients during the 2003 outbreak in Hong Kong and shows long-lasting immunity in most recovered patients. The pseudotype assay does not require handling live SARS virus; it is a useful tool to determine neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines. PMID- 15757557 TI - SARS-related perceptions in Hong Kong. AB - To understand different aspects of community responses related to severe acute respiratory syndrome (SARS), 2 population-based, random telephone surveys were conducted in June 2003 and January 2004 in Hong Kong. More than 70% of respondents would avoid visiting hospitals or mainland China to avoid contracting SARS. Most respondents believed that SARS could be transmitted through droplets, fomites, sewage, and animals. More than 90% believed that public health measures were efficacious means of prevention; 40.4% believed that SARS would resurge in Hong Kong; and approximately equals 70% would then wear masks in public places. High percentages of respondents felt helpless, horrified, and apprehensive because of SARS. Approximately 16% showed signs of posttraumatic symptoms, and approximately equals 40% perceived increased stress in family or work settings. The general public in Hong Kong has been very vigilant about SARS but needs to be more psychologically prepared to face a resurgence of the epidemic. PMID- 15757558 TI - West Nile virus risk assessment and the bridge vector paradigm. AB - In the northeast United States, control of West Nile virus (WNV) vectors has been unfocused because of a lack of accurate knowledge about the roles different mosquitoes play in WNV transmission. We analyzed the risk posed by 10 species of mosquitoes for transmitting WNV to humans by using a novel risk-assessment measure that combines information on the abundance, infection prevalence, vector competence, and biting behavior of vectors. This analysis suggests that 2 species (Culex pipiens L. and Cx. restuans Theobald [Diptera: Cilicidae]) not previously considered important in transmitting WNV to humans may be responsible for up to 80% of human WNV infections in this region. This finding suggests that control efforts should be focused on these species which may reduce effects on nontarget wetland organisms. Our risk measure has broad applicability to other regions and diseases and can be adapted for use as a predictive tool of future human WNV infections. PMID- 15757559 TI - Methicillin-resistant Staphylococcus aureus in horses and horse personnel, 2000 2002. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infection was identified in 2 horses treated at a veterinary hospital in 2000, prompting a study of colonization rates of horses and associated persons. Seventy-nine horses and 27 persons colonized or infected with MRSA were identified from October 2000 to November 2002; most isolations occurred in a 3-month period in 2002. Twenty-seven (34%) of the equine isolates were from the veterinary hospital, while 41 (51%) were from 1 thoroughbred farm in Ontario. Seventeen (63%) of 27 human isolates were from the veterinary hospital, and 8 (30%) were from the thoroughbred farm. Thirteen (16%) horses and 1 (4%) person were clinically infected. Ninety-six percent of equine and 93% of human isolates were subtypes of Canadian epidemic MRSA-5, spa type 7 and possessed SCCmecIV. All tested isolates from clinical infections were negative for the Panton-Valentine leukocidin genes. Equine MRSA infection may be an important emerging zoonotic and veterinary disease. PMID- 15757560 TI - Malaria risk in travelers. AB - Imported malaria has been an increasing problem in several Western countries in the last 2 decades. To calculate the risk factors of age, sex, and travel destination in Swedish travelers, we used data from the routine reporting system for malaria (mixture of patients with and without adequate prophylaxis), a database on travel patterns, and in-flight or visa data on Swedish travelers of 1997 to 2003. The crude risk for travelers varied from 1 per 100,000 travelers to Central America and the Caribbean to 357 per 100,000 in central Africa. Travelers to East Africa had the highest adjusted odds ratio (OR = 341, 95% confidence intervals [CI] 134-886) for being reported with malaria, closely followed by travelers to central Africa and West Africa. Male travelers as well as children <1-6 years of age had a higher risk of being reported with malaria (OR = 1.7, 95% CI 1.3-2.3 and OR = 4.8, 95% CI 1.5-14.8) than women and other age groups. PMID- 15757561 TI - Notifiable disease surveillance and practicing physicians. AB - Primary care physicians in Germany are essential participants in infectious disease surveillance through mandatory reporting. Feedback on such surveillance should reflect the needs and attitudes of these physicians. These issues were investigated in a questionnaire survey among 8,550 randomly sampled physicians in Germany in 2001. Of the 1,320 respondents, 59.3% claimed not to have received any feedback on infectious disease surveillance, and 3.7% perceived feedback as not important. Logistic regression analysis showed that physicians in the former East Germany were 2.2 times more likely to have received feedback than those in the former West Germany. Physicians preferred to receive occasional reports (e.g., in case of outbreaks, 31.6%) as opposed to actively having to search for constantly updated information on the Internet (7.8%). The preferred formats were fax (31.7%), mail (30.9%), and the official organ of the German Medical Association (Deutsches Arzteblatt) (30.5%). Feedback of surveillance data to physicians should be delivered through occasional nonelectronic reports on current issues of local public health importance. PMID- 15757562 TI - SARS-associated coronavirus transmitted from human to pig. AB - Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) was isolated from a pig during a survey for possible routes of viral transmission after a SARS epidemic. Sequence and epidemiology analyses suggested that the pig was infected by a SARS-CoV of human origin. PMID- 15757563 TI - Mimivirus in pneumonia patients. AB - Mimivirus, the largest virus known to date, is an amebal pathogen-like Legionella sp. When Mimivirus was used as an antigen in a microimmunofluorescense assay, seroconversion was found in patients with both community- and hospital-acquired pneumonia. Mimivirus DNA was found in respiratory samples of a patient with hospital-acquired pneumonia. PMID- 15757564 TI - Methicillin-resistant Staphylococcus aureus in neonatal intensive care unit. AB - A neonatal intensive care unit outbreak was caused by a strain of methicillin resistant Staphylococcus aureus previously found in the community (ST45-MRSA-IV). Fifteen infected neonates were identified, 2 of whom died. This outbreak illustrates how a rare community pathogen can rapidly spread through nosocomial transmission. PMID- 15757565 TI - Inquilinus limosus in patients with cystic fibrosis, Germany. AB - We identified Inquilinus limosus, a recently described ?-proteobacterium, in sputum of 2 patients with cystic fibrosis whose respiratory tracts were persistently colonized for >9 months. We present data on the epidemiology, antimicrobial susceptibility, and molecular characteristics of I. limosus. PMID- 15757566 TI - Fever screening at airports and imported dengue. AB - Airport fever screening in Taiwan, July 2003-June 2004, identified 40 confirmed dengue cases. Results obtained by capture immunoglobulin (Ig) M and IgG enzyme linked immunoassay, real time 1-step polymerase chain reaction, and virus isolation showed that 33 (82.5%) of 40 patients were viremic. Airport fever screening can thus quickly identify imported dengue cases. PMID- 15757567 TI - Rumor surveillance and avian influenza H5N1. AB - We describe the enhanced rumor surveillance during the avian influenza H5N1 outbreak in 2004. The World Health Organization's Western Pacific Regional Office identified 40 rumors; 9 were verified to be true. Rumor surveillance informed immediate public health action and prevented unnecessary and costly responses. PMID- 15757569 TI - Japanese encephalitis virus in meningitis patients, Japan. AB - Cerebrospinal fluid specimens from 57 patients diagnosed with meningitis were tested for Japanese encephalitis virus. Total RNA was extracted from the specimens and amplified. Two products had highest homology with Nakayama strain and 2 with Ishikawa strain. Results suggest that Japanese encephalitis virus causes some aseptic meningitis in Japan. PMID- 15757568 TI - Human metapneumovirus RNA in encephalitis patient. AB - We describe a fatal case of encephalitis that might be correlated with primary human metapneumovirus (HMPV) encephalitis. Postmortem HMPV RNA was detected in brain and lung tissue samples from the patient. Furthermore, HMPV RNA was found in culture fluids from cells coincubated with lung tissue. PMID- 15757570 TI - Identifying relapsing fever Borrelia, Senegal. AB - We describe a nested polymerase chain reaction for the identification of Borrelia species from serum of patients with unidentified fevers. This technique, based on single nucleotide polymorphisms of the 16S ribosomal RNA gene, was used to test blood samples from 7,750 patients, 33 of whom were diagnosed with spirochete infections. Borrelia crocidurae was the only species identified. PMID- 15757572 TI - Pythiosis in Africa. AB - We report the first case of pythiosis from Africa in an 8-month-old dog with a chronic and ulcerative cutaneous lesion. The etiologic agent belonged to the genus Pythium. Phylogenetic analysis placed the isolate in a sister group to the other P. insidiosum strains. However, the isolate may belong to a new Pythium species. PMID- 15757571 TI - Babesia microti, upstate New York. AB - Five cases of human babesiosis were reported in the Lower Hudson Valley Region of New York State in 2001. An investigation to determine if Babesia microti was present in local Ixodes scapularis ticks yielded 5 positive pools in 123 pools tested, the first detection of B. microti from field-collected I. scapularis in upstate New York. PMID- 15757573 TI - Hepatitis E infections, Victoria, Australia. AB - In the first half of 2004, acute hepatitis E virus infections diagnosed in Victoria, Australia, increased 7-fold. Of the interviewed patients with highly reactive serologic results, 90% reported recent clinically compatible illness and overseas travel. The increase is compared with a background of exposure in countries in which hepatitis E is endemic. PMID- 15757574 TI - Concomitant tickborne encephalitis and human granulocytic ehrlichiosis. AB - We report a patient with febrile illness and epidemiologic and clinical findings consistent with human granulocytic ehrlichiosis and tickborne encephalitis, in whom infection with Anaplasma phagocytophilum was demonstrated by polymerase chain reaction and seroconversion. Tickborne encephalitis virus infection was established by serum immunoglobulin (Ig) M and IgG antibodies. PMID- 15757575 TI - Noroviruses in archival samples. AB - Application of recent techniques to detect current pathogens in archival effluent samples collected and concentrated in 1987 lead to the characterization of norovirus GGII.6 Seacroft, unrecognized until 1990 in a clinical sample. Retrospective studies will likely increase our knowledge about waterborne transmission of emerging pathogens. PMID- 15757577 TI - Everybody else is: Networks, power laws and peer contagion in the aggressive recess behavior of elementary school boys. AB - This paper develops a simple random network model of peer contagion in aggressive behavior among inner-city elementary school boys during recess periods. The model predicts a distribution of aggressive behaviors per recess period with a power law tail beginning at two aggressive behaviors and having a slope of approximately -1.5. Comparison of these values with values derived from observations of aggressive behaviors during recess at an inner-city elementary school provides empirical support for the model. These results suggest that fluctuations in aggressive behaviors during recess arise from the interactions between students, rather than from variations in the behavior of individual students. The results therefore support those interventions that aim to change the pattern of interaction between students. PMID- 15757578 TI - Origins of group coordination: nonlinear dynamics and the role of verbalization. AB - Coordination occurs when two or more people do the same or complimentary tasks simultaneously; its explanation game theory, nonlinear dynamics, and implicit learning theory. In the experiment, 12 four-person groups were allowed to discuss the coordination (card game) task while performing it; 12 other groups worked nonverbally. One to three group members were replaced during the game. Split-plot ANOVA showed that verbalizing groups performed better than nonverbalizing groups overall and showed more acute coordination learning curves, but verbalization did not compensate for the replacement of personnel. Groups that changed one or two players showed positive coordination transfer, but groups that changed three players did not. Nonlinear regression for temporal dynamics within verbalizing and nonverbalizing groups showed asymptotic stability for initial coordination learning and transfer to a difficult rule, a chaotic function when replacements were introduced, and asymptotic stability again when the team with replacements switched to the difficult rule. PMID- 15757580 TI - [Oral microbiology in the era of genomics and proteomics]. PMID- 15757581 TI - [Pharmacokinetic/pharmacodynamic analysis of antibiotic therapy in dentistry and stomatology]. AB - INTRODUCTION: This study evaluates the efficacy of various antimicrobial treatments for orofacial infections on the basis of pharmacokinetic/pharmacodynamic (PK/PD) criteria. METHODS: A complete a literature search was undertaken to establish the MIC90 values of the five microorganisms most frequently isolated in odontogenic infections and the pharmacokinetic parameters of 13 antibiotics used in these infections. Pharmacokinetic simulations were then carried out with mean population parameters and efficacy indexes were calculated for the 47 treatment regimens analyzed. For drugs showing time-dependent antibacterial killing, the time above MIC (t > MIC) was calculated. For drugs with concentration-dependent bactericidal activity, the AUC/MIC was calculated. RESULTS: Amoxicillin-clavulanic (500 mg/8 h or 1000 mg/12 h) and clindamycin (300 mg/6 h) in the time-dependent killing group and moxifloxacin (400 mg/24 h) in the concentration-dependent group showed adequate efficacy indexes against the five pathogens considered to be the most commonly implicated in odontogenic infections. The spiramycin plus metronidazole combination, present in the commercial formulation Rhodogyl, did not reach satisfactory PK/PD indexes. CONCLUSION: PK/PD indexes, which are useful predictors of the potential efficacy of antibacterial therapy, were used with ontogenic infections in the present study. The PK/PD simulations showed that amoxicillin-clavulanic, clindamycin and moxifloxacin were the most suitable antibiotics for this kind of infection. Clinical trials are required to confirm that this methodology is useful in these pathologic processes. PMID- 15757582 TI - [Enzymatic resistance to beta lactam antibiotics within the genus Proteus and evaluation of Proteus mirabilis phenotypes and genotypes for resistance to third- and fourth-generation cephalosporins]. AB - INTRODUCTION: The aim of this study was to evaluate betalactam resistance within the genus Proteus and characterize the betalactamases responsible for this resistance. METHODS: We analyzed 99 strains (87, P. mirabilis; 10 P. vulgaris, and 2, P. penneri) isolated from patients at one University Hospital. Antibiotic susceptibility tests were performed according to NCCLS recommendations. Presence of extended spectrum betalactamases (ESBL) was inferred by both double disk diffusion tests and minimum inhibitory concentration (MIC) of third and fourth generation cephalosporins alone and in the presence of clavulanic acid. Isoelectric points (pI) of the enzymes were estimated by isoelectrofocusing and the presence of the encoding genes was confirmed by polymerase chain reaction (PCR). RESULTS: A broad spectrum betalactamase could be detected in those isolates (28%) resistant to penicillin and first generation cephalosporins while CTX-M-2 enzyme could be detected in P. mirabilis isolates resistant to third and fourth generation cephalosporins (18%). One of the P. vulgaris displayed reduced susceptibility to cefotaxime due to an enzyme of pI 7.4, while resistance to cefotaxime in one P. penneri was related to an enzyme of pI 6.8. Both enzymes were active on cefotaxime (1,000 mg/l) in the iodometric assay. CONCLUSION: The broad extended spectrum betalactamase within genus Proteus was TEM-1, while CTX-M 2 was the ESBL responsible for the third and fourth generation cephalosporins in P. mirabilis. In P. vulgaris and P. penneri this resistance was associated with the hyperproduction of the chromosomal encoded betalactamase. PMID- 15757583 TI - [Serotype and phage type distribution of human Salmonella strains isolated in Spain, 1997-2001]. AB - INTRODUCTION: Salmonellosis is one of the most frequent causes of gastroenteritis in Spain. Serotyping is the gold standard epidemiological marker for subdividing Salmonella spp. strains. A small number of serotypes are very frequently isolated, reducing the discriminatory power of serotyping. Thus, to increase our knowledge of Salmonella spp. epidemiology, additional epidemiological markers, such as phage typing, should be used for this purpose. METHODS: Salmonella spp. strains of human origin sent to the Laboratorio Nacional de Referencia de Salmonella y Shigella (LNRSSE, Spanish Reference Laboratory for Salmonella and Shigella) between 1997 and 2001 were serotyped using conventional agglutination methods, and Enteritidis, Typhimurium, Hadar, Virchow and Typhi serotypes were additionally phage typed according to internationally-developed schemes. RESULTS: A total of 30,856 Salmonella spp. strains, isolated in the majority of Spanish Autonomous Communities, were analyzed. Enteritidis (51%) and Typhimurium (24%) were the most frequently isolated serotypes. The following were the most frequent serotype/phage type combinations: Enteritidis/PT1 (18%), Enteritidis/PT4 (15%), Enteritidis/PT6a (5%), Typhimurium/DT104 (5%) and Enteritidis/PT6 (3%). The serotype Enteritidis/PT1 showed the greatest increase over the period studied, from 11.61% in 1997 to 24.74% in 2001. CONCLUSIONS: A hierarchical typing approach for Salmonella spp., using serotyping coupled with phage typing allowed a higher level of discrimination among Salmonella serotypes. Application of this approach in epidemiological studies could be highly useful for early characterization of related strains. PMID- 15757585 TI - [Consolidation of a Staphylococcus aureus multiresistant clone not related to the Iberian in a hospital from Mallorca]. AB - INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) was seldom isolated in our hospital until the first outbreak in 1999. A recently documented increase in antibiotic multiresistance in MRSA strains in our setting prompted the design of this molecular epidemiology study to investigate the basis for this tendency. METHODS: All MRSA isolates from clinical samples of patients admitted from July 2002 to June 2003 were studied. Susceptibility testing was performed by disk diffusion. Clonal relatedness of MRSA isolates was determined by pulsed field gel electrophoresis (PFGE). Results were compared with data from MRSA isolates from patients admitted to the hospital in 1999-2000. RESULTS: MRSA was isolated in 110 patients (30% of patients with S. aureus-positive cultures). PFGE detected three major clones (in 93% of patients), all of which had been present in 1999-2000, although in different proportions. Whereas the predominant clone in 1999 was clone A (clone A 63%, clone B 20%), clone B was now found to predominate (clone B 58%, clone A 19%). None of these major clones were related to the five pandemic clones, including the Iberian clone, but two of them seemed to be related to the two most prevalent clones in Spain at this time. The new predominant clone was more resistant than the others, and showed uniform resistance to ciprofloxacin, erythromycin, clindamycin, and gentamicin. CONCLUSION: In recent years, a formerly predominant MRSA clone has been replaced by a multiresistant S. aureus clone that is unrelated to the Iberian clone. PMID- 15757584 TI - [Blood cultures in the emergency department]. AB - INTRODUCTION: Culture of emergency room blood samples is common practice, but open to controversy. As compared to other emergency tests, blood collection requires twice as much time and needs a refined technique to avoid contamination, and the study has no immediate diagnostic utility. METHODS: This prospective study includes consecutive adult patients with positive emergency room blood cultures. We analyzed the diagnostic sensitivity and contamination rate of the cultures and the etiology, clinical features and prognosis of the bacteremias encountered. RESULTS: During the study period, 5.2 blood cultures were indicated per 1000 patients attended in the emergency room. The diagnostic yield (positive blood cultures/total cultures) was 20% and the contamination rate (contaminated blood cultures/total cultures) was 1%. The incidence of bacteriemia was 0.99 episodes per 1000 patients attended in the emergency room and 10.3 episode per 1000 hospitalized patients. Gram-negative bacteria predominated (57%). Sepsis was the most frequent clinical manifestation (50%), followed by severe sepsis (40%) and septic shock (10%). Mortality was 22%. Diabetes mellitus and severe sepsis/septic shock were independent factors associated with mortality. CONCLUSIONS: Diagnostic performance and quality of emergency room blood cultures was high. The predominant etiology was gram-negative bacteria. Patients had a severe clinical presentation. Diabetes mellitus and severe sepsis and/or septic shock were independent prognostic factors of mortality. PMID- 15757586 TI - [Clinical and epidemiological study of a series of HIV-infected patients over 50 years old]. AB - INTRODUCTION: The epidemiology of human immunodeficiency virus (HIV) infection has changed in recent years. Cases in persons over the age of 50 have increased, and the most frequent mode of transmission is sexual contact. The objective of this study is to analyse the epidemiological, clinical and evolution characteristics of a clinical series of HIV-infected patients over 50 years old at the time of diagnosis. METHODS: 165 HIV-infected patients over the age of 50, attended at Hospital Clinic (Barcelona) and Corporacio Parc Tauli (Sabadell) during the period of 1985 to 2001, were studied. RESULTS: Among the total, 81% of the patients were men, mean age at the time of diagnosis was 58.5 years, and 81.8% had acquired the disease by sexual contact. The median initial CD4 T cell count was 216 cells/microl. Initial viral load was 1,000-100,000 copies/ml in 45.2% of the patients, whereas 52.3% had > 100,000 copies/ml. At the time of diagnosis, 30.9% had an AIDS-defining disease. The main opportunistic diseases were pulmonary tuberculosis, Kaposi's sarcoma, P. jiroveci (before carinii) pneumonia and non-Hodgkin lymphoma. Mortality due to AIDS was 32.7%. CONCLUSIONS: Subjects over 50 years old diagnosed with HIV-infection were predominantly men, who acquired the infection by sexual contact. A high percentage of patients were diagnosed with the development of an opportunistic disease. PMID- 15757587 TI - [Study of resistance using the TRUGENE HIV-1 genotyping system and analysis of agreement between rule-based algorithms and virtual phenotyping]. AB - INTRODUCTION: This study attempts to describe the results obtained in the HIV resistance study performed with clinical samples obtained from patients receiving "HAART" therapy and to compare the results using different interpretation algorithms. METHODS: 397 samples have been analysed (TRUGENE HIV-1 GENOTYPING kit). The results were interpreted with the algorithms Visible Genetics and Retrogram. The concordance interalgorithm was done for 105 of these samples, including the virtual phenotype interpretation. RESULTS: The samples corresponded to multi regimen failure (39%), first and second failure (30.7% and 27.1% respectively). A 27.6% of the samples were wild type. The more frequent mutations to the ITIAN were T215Y/F (37.2%) and M184V (32.9%) following by other NAMS. The 69 insertion and Q151M complex had low representativity. For the ITINN K103N (25.8%), Y181C (11.2%) and G190A (10.9%). For the IP: key mutations L90M (26.1%), M46I (18.1%) and V82AFTS (12.9%); and accessory mutations L63P (50.5%), A71V (27.2%), L10I (25.2%) and M36I (19.2%). Low correlation was observed between interpretation systems, mainly for ITIAN and IP, being the virtual phenotype more flexible in the assignation of resistance. CONCLUSIONS: The requests for HIV resistance testing were similar for the three groups of patients. Many of the failures were the consequence of a poor adherence to the therapy. The mutation pattern found corresponded with the "TARGA" therapy. The low correlation found between interpretation systems, makes necessary the elaboration of a consensus algorithm. PMID- 15757588 TI - [Regulation of transcription in different HIV-1 subtypes]. AB - Transcriptional activation of HIV-1 gene expression is controlled in part by the interaction of viral and cellular transcription factors with the HIV-1 long terminal repeat (LTR) sequences. LTR variability among different HIV-1 subtypes could affect LTR binding of either cellular or viral elements, influencing the transcription level. This effect, in turn, may have consequences on the biology of the different HIV-1 clades and on disease progression. In some circumstances, a relationship between replication capacity in vitro and changes in binding sequences for transcription factors located at the LTR has been proven. PMID- 15757589 TI - [Rickettsioses]. AB - Species of the genus Rickettsia are small, obligate intracelular, gramnegative bacteria, many of which are considered nowadays a paradigm of emergent pathogens. With the exception of R. prowazekii, they are maintained in the natural environment through a cycle involving different hosts (mainly mammals), and arthropod vectors (in general ticks, and fleas); humans are affected only by incidental transmission due to arthropod bites. The common pathogenesis of these diseases lie on the predominantly infection of endotelial cells, that determines the development of multisistemic small vessel vasculitis, which may affect lungs (interstitial pneumonitis), heart (miopericarditis), skin (rash), central nervous system (meningoencephalitis), as well as liver, and kidneys. They are classified in two groups: spotted-fever group, and typhus group rickettsia. In Spain the most prevalent rickettsioses of both groups are mediterranean spotted fever (caused by R. conorii), and murine typhus (caused by R. typhi), respectively. This review focuses mainly in these two diseases, and also in other rickettsioses of interest due to their recently emergence or reemergence (R. slovaca, R. africae, R. prowazekii, R. felis), or to their high incidence in other areas (R. rickettsii, Orientia tsutsugamushi). PMID- 15757590 TI - [Sudden neurological deterioration in a patient addicted to intravenous drugs]. PMID- 15757591 TI - [Secondary syphilis in an HIV patient]. PMID- 15757592 TI - [Ulcerated suppurating skin lesion of the neck with slow progression in an 83 year-old woman]. PMID- 15757593 TI - [Liver abscess due to a native amoeba: a clinical case study and literature review]. PMID- 15757594 TI - [Urinary tract infection caused by bacteria of the genus Aeromonas]. PMID- 15757595 TI - Impact of obesity on revascularization and restenosis rates after bare-metal and drug-eluting stent implantation (from the TAXUS-IV trial). AB - The effect of obesity on repeat coronary revascularization and restenosis in patients who undergo stent implantation has not been reported. We therefore examined the database from the multicenter randomized TAXUS-IV trial to determine the effect of body mass index (BMI) on outcomes after bare-metal and drug-eluting stent implantation. In TAXUS-IV, patients were randomized to receive a slow release, polymer-based, paclitaxel-eluting stent or a bare-metal stent. Outcomes were stratified by baseline BMI. Of the 1,307 randomized patients who had documented BMI, 233 (17.8%) had normal weight (BMI <25 kg/m2), 531 (40.6%) were overweight (BMI < or =25 to 30 kg/m2), and 543 (41.5%) were obese (BMI > or =30 kg/m2). Patients who had been assigned to receive bare-metal stents and were overweight and obese compared with those who had normal weight had higher rates of 9-month binary restenosis (29.2% and 30.5% vs 9.3%, respectively; p = 0.01) and 1-year major adverse cardiac events (20.8% and 23.2% vs 11.1%, respectively; p = 0.02), whereas rates of these events did not differ significantly among those who received a paclitaxel-eluting stent (7.6% and 9.3% vs 4.9%, respectively for binary restenosis; p = 0.65; 11.3% and 10.4% vs 10.1%, respectively; p = 0.82 for major adverse cardiac events). By multivariate analysis, BMI > or =30.0 kg/m2 independently predicted binary restenosis (hazard ratio 4.26, p = 0.005), 1-year target vessel revascularization (hazard ratio 1.95, p = 0.04), and major adverse cardiac events (hazard ratio 1.95, p = 0.004) in patients who received bare-metal stents but not paclitaxel-eluting stents. In conclusion, obesity is an important risk factor for clinical and angiographic restenosis and for composite major adverse cardiac events in patients who receive bare-metal stents. Paclitaxel eluting stents attenuate the increased risk associated with obesity, such that the intermediate-term prognosis after percutaneous coronary intervention is independent of weight. PMID- 15757596 TI - Effectiveness and safety of bivalirudin during percutaneous coronary intervention in a single medical center. AB - A recent large-scale, randomized trial demonstrated the noninferiority of a strategy of bivalirudin with provisional glycoprotein (GP) IIb/IIIa inhibition compared with routine GP IIb/IIIa inhibition. There is a paucity of outcome data with bivalirudin use in the setting of real-world experience. We evaluated 6,996 patients who underwent percutaneous coronary intervention between January 2001 and December 2004 to compare early and late outcomes with a bivalirudin-based antithrombotic regimen with those with a heparin-based regimen. Propensity adjustment was performed to correct for baseline differences in patient characteristics. Bivalirudin-based therapy was used in 1,070 patients, heparin only in 801 patients, and heparin plus GP IIb/IIIa inhibitors in 5,125 patients. Compared with patients who received heparin or those who received heparin plus GP IIb/IIIa inhibitors, patients who received bivalirudin had lower incidences of bleeding (blood transfusion rate 1.7% vs 4.0%, p <0.001) and periprocedural myonecrosis (creatine kinase-MB >5 times the upper limit of normal 2.7% vs 4.3%, p = 0.016). Differences in bleeding end points remained significant after adjusting for the propensity to receive bivalirudin, but there was no difference in ischemic events. There was no difference in unadjusted long-term survival rate (log-rank test p = 0.46, total number of deaths 412, mean follow-up 17 months) or in propensity-adjusted long-term survival rate (hazard ratio 1.37, 95% confidence interval 0.90 to 2.08, p = 0.14). Compared with heparin with or without GP IIb/IIIa inhibition, the use of bivalirudin in a large consecutive patient registry at a tertiary care center was associated with fewer bleeding events and no evident increase in the incidence of ischemic complications. PMID- 15757597 TI - Characterization of hibernating myocardium with NOGA electroanatomic endocardial mapping. AB - Because the terms "hibernation" and "viability" are not interchangeable, the recognition of hibernating myocardium within viable segments remained elusive for NOGA electroanatomic endocardial mapping. The aim of the present study was to determine the characteristics of hibernating myocardium in NOGA mapping. Baseline and follow-up endocardial mapping, thallium-201 myocardial perfusion scintigraphy at rest, and contrast ventriculography were performed in 28 patients who had proved viable myocardium before and 7.3 +/- 2.5 months after percutaneous coronary intervention. Significantly improved regional wall motion in the revascularized territory (region of interest) was confirmed in 9 patients (group 1) at follow-up (from -2.11 +/- 0.87 to -1.48 +/- 0.43 SD/chord, p <0.05), whereas no change in regional wall motion was observed in 19 patients (group 2; from -2.56 +/- 0.88 to -2.79 +/- 0.91 SD/chord). Average normalized thallium uptake at rest increased significantly in groups 1 and 2 after revascularization. A trend toward increased unipolar voltages in the region of interest was observed in group 1 at follow-up (from 10.6 +/- 3.5 to 11.7 +/- 4.0 mV, p = 0.073), whereas no change was observed in group 2 (from 8.7 +/- 4.4 to 8.9 +/- 3.8 mV). A significant increase in local linear shortening was measured only in group 1 (from 7.5 +/- 5.2% to 10.3 +/- 3.9%, p <0.05). Hibernating myocardial segments exhibited significantly higher unipolar voltages and late thallium uptake at rest at baseline. Receiver-operator characteristic analysis showed a mean unipolar voltage of 9.0 mV (predictive accuracy 0.708, common sensitivity and specificity 72%) in the region of interest for prediction of functional recovery. In conclusion, for characterizing the hibernating myocardium within viable segments, NOGA endocardial mapping offers on-line guidance for percutaneous coronary and noncoronary myocardial revascularization. PMID- 15757598 TI - Plasma asymmetric dimethylarginine concentrations in newly diagnosed patients with acute myocardial infarction or unstable angina pectoris during two weeks of medical treatment. AB - A high concentration of plasma asymmetric dimethylarginine (ADMA) has been associated with several risk factors for atherosclerosis, and this may increase the risk for acute coronary syndromes (ACSs). We measured plasma ADMA concentrations in patients who had newly diagnosed ACS (n = 48), and we followed the changes in ADMA concentrations during these patients' short-term medical therapy, which included various combination of drugs with or without percutaneous coronary interventions according to the needs of each patient. Concentrations of plasma ADMA were found to be high in patients who had ACS compared with 48 age matched healthy control subjects (3.13 +/- 0.85 vs 1.57 +/- 0.85 mumol/L, p <0.0001). Follow-up measurements of ADMA showed dramatic decreases in plasma ADMA concentrations over 2 weeks of medical therapy for ACS (from 3.27 +/- 0.87 to 1.52 +/- 0.47 mumol/L, p <0.0001). Plasma ADMA at baseline showed a significant positive correlation with serum C-reactive protein and plasma insulin and a significant negative correlation with serum levels of high-density lipoprotein and plasma alpha-tocopherol. During therapy, changes in plasma ADMA concentrations were significantly correlated with changes in the ratio of total cholesterol to high-density lipoprotein cholesterol and in serum C-reactive protein concentrations but not with changes in insulin levels. This study provides the first evidence that plasma ADMA concentrations are significantly high in patients who have ACS and that ADMA concentrations rapidly decrease after short-term medical therapy. PMID- 15757599 TI - Validation of peak exercise oxygen consumption and the Heart Failure Survival Score for serial risk stratification in advanced heart failure. AB - The Heart Failure Survival Score (HFSS) and peak exercise oxygen consumption (VO2) accurately assess mortality in ambulatory patients who have advanced heart failure and are referred for initial cardiac transplant evaluation. We investigated the prognostic value of the HFSS and peak VO2 when applied serially to these patients. This study included 227 adults (mean age +/- SD 52 +/- 10 years old) who presented for reevaluation >60 days after initial evaluation (352 +/- 238 days). The HFSS was determined from mean arterial blood pressure, heart rate, left ventricular ejection fraction, serum sodium, peak VO2, heart failure etiology, and width of QRS complex. Survival without reevaluation, United Network of Organ Sharing 1 transplant, or left ventricular assist device was determined by the Kaplan-Meier method with censoring at United Network of Organ Sharing 2 transplant. Survival differed by HFSS stratum (p <0.001) and by peak VO2 stratum (p <0.001). Patients whose HFSS or peak VO2 deteriorated from low risk to medium or high risk had lower survival rates than did patients whose values remained at low risk (p <0.01 and p <0.001, respectively). Patients who started at medium or high risk and improved to low risk tended to have higher survival rates than those who remained medium or high risk (p = 0.06 and p <0.16, respectively). Patients who improved to low risk had a 1-year survival rate of 72% for HFSS and peak VO2. However, patients who improved to low risk and were treated with beta blockers had a 1-year survival rate (89% for HFSS and 83% for peak VO2) comparable to that after transplant (84%). Peak VO2 and the HFSS can be successfully used for serial evaluation of mortality risk in ambulatory patients who have advanced heart failure. PMID- 15757600 TI - Prevalence of valve calcium and association of valve calcium with coronary artery disease, atherosclerotic vascular disease, and all-cause mortality in 137 patients undergoing hemodialysis for chronic renal failure. AB - Of 137 patients (mean age 63 years) who underwent hemodialysis for chronic renal failure, 65 (47%) had mitral valve calcium, mitral annular calcium, or aortic valve calcium. Thirty-eight of 65 patients (59%) who had valve calcium died at 3.5-year follow-up versus 21 of 72 patients (29%) who did not have valve calcium and who died at 4.3-year follow-up (p = 0.0005). PMID- 15757601 TI - Incidence of antiplatelet factor 4/heparin antibody induction in patients undergoing percutaneous coronary revascularization. AB - The incidence of antiplatelet factor-4/heparin antibody formation in patients who receive contemporary doses of unfractionated heparin in the setting of percutaneous coronary revascularization is unknown. Also unknown is the ability of these antibodies to activate platelets or adversely affect clinical outcome in the absence of clinically recognized heparin-induced thrombocytopenia. To address these questions, we serially measured antiplatelet factor-4/heparin antibody levels and performed serotonin release assays in patients who underwent percutaneous coronary intervention. Correlations were then made across antibody induction, heparin exposure, and clinical outcome at 6 months. PMID- 15757602 TI - Comparison of C-reactive protein levels after coronary stenting with bare metal versus sirolimus-eluting stents. AB - We evaluated C-reactive protein increases after implantation of bare metal stents in 200 patients and sirolimus-eluting stents in 100 patients. The magnitude of change in C-reactive protein was comparable between groups. Clinical follow-up showed a relation between the postprocedural C-reactive protein increase and outcome that was significant in the bare metal stent group, which accounted for the most of events, but not in the sirolimus-eluting stent group. PMID- 15757604 TI - Risks of noncardiac surgery after coronary stenting. AB - An increased risk of major complications for noncardiac surgery after coronary stenting has been suggested. We retrospectively reviewed all cases of coronary stents from 1999 to 2003 with subsequent surgery to assess major adverse cardiovascular events (MACEs), including myocardial infarction, stent thrombosis, major bleeding, and death. Among the 56 patients identified, 8 developed MACEs; 38% underwent surgery < or =14 days after stenting, and 62% underwent surgery 15 to 42 days after stenting. No patient developed MACEs if surgery occurred >42 days after stenting. Among patients who developed MACEs, 77% of surgeries were elective, 19% were urgent, and only 4% were emergency. Noncardiac surgery 6 weeks after coronary stenting is associated with a high risk of MACEs. PMID- 15757603 TI - A novel mechanism explaining early lumen loss following balloon angioplasty for the treatment of in-stent restenosis. AB - We performed serial intravascular ultrasound analysis in patients who underwent balloon dilatation for in-stent restenosis. Early lumen loss was detected by intravascular ultrasound and was associated with minimal changes at the edges and at the external elastic membrane. These results on intravascular ultrasound suggest compression and decompression as the main mechanisms for early lumen loss after dilatation of in-stent restenotic lesions. PMID- 15757605 TI - In-hospital and nine-month outcome of treatment of coronary bifurcational lesions with sirolimus-eluting stent. AB - Between April 2002 and May 2004, 174 consecutive patients who underwent percutaneous coronary intervention of bifurcational lesions with sirolimus eluting stents were identified. Two strategies were used: stenting only 1 branch (group 1S, n = 57) or stenting both branches (group 2S, n = 117). The incidence of major adverse cardiac events was evaluated in the hospital and at 9-month follow-up. There were no statistically significant differences between the 2 groups with regard to the incidence of target lesion revascularization (5.4% vs 8.9%, p = 0.76), target vessel revascularization (5.4% vs 11.1%, p = 0.51), and cumulative major adverse cardiac events (18.9% vs 23.3%, p = 0.76) at 9 months. PMID- 15757606 TI - Quality of life in octagenarians after coronary artery bypass grafting. AB - Efficacy of coronary artery bypass grafting is well accepted, but its role in octogenarians remains unclear. Therefore, quality of life was evaluated in 73 consecutive octogenarians who underwent coronary artery bypass grafting (1996 to 2001) with data collected in terms of age, gender, New York Heart Association status, co-morbidities, and intra- and postoperative complications. Quality of life was evaluated by a standardized questionnaire to assess postoperative outcomes in terms of functional status (Karnofsky scale), morbidity, and mortality. PMID- 15757607 TI - Relation of elevated C-reactive protein and interleukin-6 levels to left atrial size and duration of episodes in patients with atrial fibrillation. AB - Previous studies have demonstrated inflammation to be a risk factor in patients with atrial fibrillation (AF). In this prospective study of 90 patients with persistent and permanent AF and 46 controls, we found increased C-reactive protein (CRP) and interleukin-6 levels in patients with AF compared with controls (p <0.001). Multivariate analysis revealed CRP to be an independent predictor of AF (p = 0.01). Left atrial diameter was positively related to CRP and interleukin 6 (p <0.001, R = 0.37; p <0.001, R = 0.46, respectively) and negatively related to left ventricular function. Interleukin-6 levels were positively related to AF duration before cardioversion (p = 0.02). Elevation of CRP and interleukin-6 suggest a role of inflammation in AF, and the relation of CRP and interleukin-6 to left atrial size and AF duration before cardioversion indicates that inflammation may participate in the process of atrial remodeling. PMID- 15757608 TI - Efficacy and safety of intermittent, long-term, concomitant dobutamine and levosimendan infusions in severe heart failure refractory to dobutamine alone. AB - Thirty-six consecutive patients in New York Heart Association functional class IV, who were resistant to 24-hour continuous dobutamine infusion, were treated with continuous infusions of dobutamine 10 microg/kg/min for > or =48 hours (group I, n = 18), followed by weekly intermittent 8-hour infusions or more often if needed. In group II (n = 18), after the initial 24-hour infusion of dobutamine, a 24-hour levosimendan infusion was added followed by biweekly 24 hour infusions. The addition of intermittent levosimendan infusions prolonged the survival of patients with advanced heart failure refractory to intermittent dobutamine infusions (45-day survival rates were 6% and 61% in groups I and II, respectively; p = 0.0002, log-rank test). PMID- 15757609 TI - Effect of chronic right ventricular apical pacing on left ventricular function. AB - The determinants of change in left ventricular (LV) ejection fraction (EF) over time in patients with impaired LV function at baseline have not been clearly established. Using a nuclear database to assess changes in LV function over time, we included patients with a baseline LVEF of 25% to 40% on a gated single-photon emission computed tomographic study at rest and only if second-gated photon emission computed tomography performed approximately 18 months after the initial study showed an improvement in LVEF at rest of > or =10 points or a decrease in LVEF at rest of > or =7 points. In all, 148 patients qualified for the EF increase group and 59 patients for the EF decrease group. LVEF on average increased from 33 +/- 4% to 51 +/- 8% in the EF increase group and decreased from 35 +/- 4% to 25 +/- 5% in the EF decrease group. The strongest multivariable predictor of improvement of LVEF was beta-blocker therapy (odds ratio 3.9, p = 0.002). The strongest independent predictor of LVEF decrease was the presence of a permanent right ventricular apical pacemaker (odds ratio 6.6, p = 0.002). Thus, this study identified beta-blocker therapy as the major independent predictor for improvement in LVEF of > or =10 points, whereas a permanent pacemaker (right ventricular apical pacing) was the strongest predictor of a LVEF decrease of > or =7 points. PMID- 15757610 TI - Physical determinants of systolic murmur intensity in aortic stenosis. AB - We investigated which physical parameter has the greatest impact on the perceived loudness of a systolic murmur in aortic stenosis. Loudness of murmur in aortic stenosis correlated best with peak momentum transfer--and thus with body size--so that loud murmurs predict severe disease less reliably in larger patients. PMID- 15757611 TI - Outcome of operated and unoperated adults with congenital cardiac disease lost to follow-up for more than five years. AB - Many patients with congenital cardiac disease need a regular cardiologic follow up (FU) even after successful primary treatment. Nevertheless, many of them are lost to FU. The present study verifies for the first time the outcome of adults with congenital cardiac disease lost to FU of a specialized institution for >5 years. PMID- 15757612 TI - Optimal timing for pulmonary valve replacement in adults after tetralogy of Fallot repair. AB - The timing of pulmonary valve replacement in adult patients with repaired tetralogy of Fallot remains controversial. A magnetic resonance imaging study in 17 adult patients with repaired tetralogy of Fallot reveals a statistically significant decrease in right ventricular (RV) volume (RV end-diastolic volume 163 +/- 34 to 107 +/- 26 ml/m2, p <0.001; RV end-systolic volume 109 +/- 27 to 69 +/- 22 ml/m2, p <0.001) at a mean follow-up of 21 months after pulmonary valve replacement; whereas RV systolic function remained unchanged (mean RV ejection fraction 32 +/- 7% to 34 +/- 10%, p = 0.12). In no patients with a RV end diastolic volume >170 ml/m2 or a RV end-systolic volume >85 ml/m2 before pulmonary valve replacement were RV volumes "normalized" after surgery. PMID- 15757613 TI - Catheter management of occluded superior baffle after atrial switch procedures for transposition of great vessels. AB - We present successful catheter intervention in 4 patients with total occlusion of the superior baffle after an atrial switch procedure. A transseptal needle (in addition to the transhepatic route in 1 patient with known occluded femoral veins) was used to open the occluded segments and place stents. Additionally, we report the regression of multiple systemic-to-pulmonary venous fistulas that developed as a result of the occluded baffle. PMID- 15757614 TI - Effect of triiodothyronine on gene transcription during cardiopulmonary bypass in infants with ventricular septal defect. AB - We tested the hypothesis that triiodothyronine (T3) supplementation alters gene transcription in the left ventricular myocardium of infants undergoing cardiopulmonary bypass for ventricular septal defect repair. To our knowledge, a novel heteronuclear assay demonstrated for the first time in human heart that rapid change in T3 levels altered the adenine nucleotide translocase-1 transcription rate during cardiopulmonary bypass. PMID- 15757615 TI - Effect of serum albumin on QRS wave amplitude in patients free of heart disease. AB - We studied 193 patients free of heart disease to determine the relation between QRS amplitude and serum albumin. Although there were no significant differences in echocardiographic indexes between the 2 groups, albumin (35.1 +/- 4.3 vs 40.1 +/- 3.2 g/L) and colloid osmotic pressure (21 +/- 4 vs 24 +/- 3 mm Hg) were significantly lower in patients with low voltages compared with those without. Moreover, there was a good relation (r = 0.78) between change in QRS amplitude and change in albumin concentration. PMID- 15757616 TI - Comparison of the safety and efficacy of emboli prevention devices versus platelet glycoprotein IIb/IIIa inhibition during carotid stenting. AB - Distal embolization is the main potential risk of carotid stenting, and techniques to minimize this risk are evolving. Between July 1998 and March 2002, 305 consecutive patients who underwent elective or urgent percutaneous carotid intervention at The Cleveland Clinic were prospectively followed. During this period, the clinical practice of carotid stenting evolved from the routine use of glycoprotein IIb/IIIa inhibitors (GPIs) to routine emboli-prevention device (EPD) placement. A total of 199 patients received adjunctive GPIs (91% abciximab), and 106 patients underwent the procedure with an EPD (85% filter design, 15% occlusive balloon). At 30 days, the composite end point of neurologic death, nonfatal stroke, and major bleeding, including intracranial hemorrhage, was significantly lower among patients treated with EPDs compared with those treated with GPIs (0% vs 5.1%, p = 0.02). EPDs may provide an overall safer and more effective means of neuroprotection during carotid stenting than GPIs. PMID- 15757617 TI - Evaluation of left ventricular dimensions and function in Marfan's syndrome without significant valvular regurgitation. AB - Left ventricular dimensions and systolic function were studied using echocardiography in 234 patients with Marfan's syndrome without significant valvular regurgitation. Left ventricular dimensions and systolic function were found to be normal in most patients with Marfan's syndrome. Some involvement of the left ventricle may have been present in a small group of these patients. No patients, however, fulfilled the criteria for dilated cardiomyopathy. PMID- 15757618 TI - Left ventricular function in patients with type 2 diabetes mellitus. AB - This study showed that the mean left ventricular ejection fraction, end-diastolic volume, end-systolic volume, and muscle mass are comparable in patients with type 2 diabetes mellitus to gender-matched patients who do not have diabetes mellitus, but abnormal ejection fraction is more common in men, although not in women, with diabetes mellitus than without. The ejection fraction was higher and the volumes and muscle mass were lower in women than men in the presence or absence of diabetes mellitus. PMID- 15757619 TI - Left ventricular function in children with sleep-disordered breathing. AB - Severe obstructive sleep apnea in children leads to congestive heart failure. We studied the early changes in left ventricular function across a range of severity of the disorder. A dose-dependent decrease in diastolic function with increased severity of obstructive apnea was demonstrated. PMID- 15757620 TI - Aspirin resistance and a single gene. AB - This study examined the association of single nucleotide polymorphisms in 4 candidate genes in a cohort of subjects with aspirin resistance. Aspirin resistance was significantly associated with genetic variation in the platelet surface adenosine 5-diphosphate receptor gene P2Y1. PMID- 15757621 TI - Comparison of left ventricular volumes and ejection fractions measured by three dimensional echocardiography versus by two-dimensional echocardiography and cardiac magnetic resonance in patients with various cardiomyopathies. AB - End-diastolic volume and end-systolic volume were measured in 35 consecutive patients with cardiomyopathy using 2-dimensional (2-D) and 3-dimensional (3-D) echocardiography (2, 4, and 8 planes) and cardiac magnetic resonance imaging. Three-dimensional echocardiography correlates better with magnetic resonance imaging than does 2-D echocardiography. Its accuracy improves with the increase in the number of planes used. Two-dimensional echocardiography underestimates volumes, mainly in the subgroup with an ejection fraction of <50%, whereas 3-D echocardiography does not, if enough planes are used. However, in patients with an end-diastolic volume > or =150 ml, the underestimation of 3-D echocardiography is statistically significant. Increasing the number of planes to 8 reduces this bias. Conversely, patients with an end-diastolic volume <150 ml are accurately studied with just 4 planes. PMID- 15757623 TI - Primary ventricular fibrillation cannot be buried; it never died. PMID- 15757625 TI - Atrial fibrillation, the prothrombotic state, inflammation, and gender. PMID- 15757626 TI - The impact of ethnicity on the lifetime risk of the metabolic syndrome and diabetes mellitus. PMID- 15757627 TI - Does aortic root dilatation with bicuspid aortic valves occur as a primary tissue abnormality or as a relatively benign poststenotic phenomenon? PMID- 15757628 TI - Mode of action of taurine as a neuroprotector. AB - Previously, it has been shown that taurine exerts its protective function against glutamate-induced neuronal excitotoxicity through its action in reducing glutamate-induced elevation of intracellular free calcium, [Ca2+]i. Here, we report the mechanism underlying the effect of taurine in reducing [Ca2+]i. We found that taurine inhibited glutamate-induced calcium influx through L-, P/Q-, N type voltage-gated calcium channels (VGCCs) and NMDA receptor calcium channel. Surprisingly, taurine had no effect on calcium influx through NMDA receptor calcium channel when cultured neurons were treated with NMDA in Mg2+-free medium. Since taurine was found to prevent glutamate-induced membrane depolarization, we propose that taurine protects neurons against glutamate excitotoxicity by preventing glutamate-induced membrane depolarization, probably through its effect in opening of chloride channels and, therefore, preventing the glutamate-induced increase in calcium influx and other downstream events. PMID- 15757629 TI - Calbindin D28k-containing neurons in the paratrigeminal nucleus receive convergent nociceptive information and project to nucleus of the solitary tract in rat. AB - The paratrigeminal nucleus (PTN) receives orofacial somatic and visceral afferent fibers and contains many calbindin-D28k neurons (CB-containing neurons) that project to nucleus of the solitary tract (NTS). In the present study, retrograde and transganglionic tracing methods combined with immunofluorescence histochemistry and confocal laser scanning microscopy were used. After Fluoro gold (FG) injection into the unilateral NTS, 74.4% FG-labeled neurons of ipsilateral PTN were double-labeled with CB. Furthermore, 41.0% and 32.5% FG/CB double-labeled neurons co-existed with Fos induced by nociceptive stimulation of the lips and the upper alimentary tract, respectively. In the PTN unilateral to FG injection site, 26.6% CB-LI neurons were double-labeled with PAG, 61.5% and 79.0% CB/PAG double-labeled neurons were triple-labeled with FG and Fos, and 22.9% FG/CB double-labeled neurons were triple-labeled with PAG, 84.3% FG/PAG double-labeled neurons expressed Fos induced by the upper alimentary tract stimulation. In the intact animals, 62.8% CB-LI neurons and 88.3% PAG-LI neurons co-existed with GABA(B)R, respectively. In addition, some terminals from the inferior alveolar nerve (IAN) were closely apposed to CB/Fos double-labeled or CB single-labeled neurons. These results suggested that CB-containing neurons in the PTN receive the nociceptive information converge from the orofacial area and visceral organs, and comprising the glutamatergic excitatory transmission pathway from the PTN to the NTS. This pathway might be modulated by GABA via the GABA(B) receptor. PMID- 15757630 TI - Expression of functional receptors and transmitter enzymes in cultured Muller cells. AB - Glia represents the most numerous group of nervous system cells and CNS development and function depend on glial cells. We developed a purified Muller glia culture to investigate the expression of several neurotransmitter markers on these cells, such as dopaminergic, cholinergic, GABAergic and peptidergic receptors or enzymes, based on functional assays measuring second messenger levels or Western blot for specific proteins. Purified Muller cell culture was obtained from 8-day-old (E8) embryonic chick. Glial cells cultured for 15 days (E8C15) expressed D1A and D1B receptors mRNAs, but not D1D, as detected by RT PCR. The binding of [3H]-SCH 23390 revealed an amount of expressed receptors around 40 fmol/mg protein. Dopamine (100 microM), PACAP (50 nM) and forskolin (10 microM) induced a 50-, 30- and 40-fold cAMP accumulation on glial cells, respectively, but not ip3 production. The dopamine-promoted cAMP accumulation was blocked by 2 microM SCH 23390. Carbachol stimulated a 3-fold ip3 accumulation. Western blot analysis also revealed the expression of tyrosine hydroxylase, L dopa decarboxylase, PAC1 receptor, GAD67 and beta2-nicotinic receptor subunit by these cells. These results indicate that several components of neurotransmitter signaling and metabolism are found in cultured Muller cells. PMID- 15757631 TI - The distribution of neural nitric oxide synthase-positive cerebrospinal fluid contacting neurons in the third ventricular wall of male rats and coexistence with vasopressin or oxytocin. AB - The detailed distribution of neural nitric oxide synthase (nNOS)-positive cerebrospinal fluid-contacting neurons (CSF-CN) was studied in the wall of the third ventricle of rats by anti-nNOS immunohistochemistry. The coexistence of nNOS and 8-arginine vasopressin (AVP) or oxytocin (OT) was also investigated in the CSF-CN using double labeling immunohistochemistry. The results demonstrated a widespread occurrence of nNOS-CSF-CN throughout the wall of the hypothalamic third ventricle. The vast majority of nNOS-CSF-CN cell bodies were of magnocellular type, commonly classified as oval, fusiform, multipolar, and inverted pear shape. These cell bodies were located in the ependyma, the subependyma, or the parenchyma, and their processes inserted in the ependymal layer or directly contacted with the CSF space. Electron microscopy demonstrated many nNOS-immunoreactive somas, dendrites, and/or axons that were situated at the subependyma, the ependyma, or the supraependyma. Generally, the distribution of OT-CSF-CN in the third ventricular wall was similar to the nNOS-CSF-CN and the ratio of NOS/OT co-expression was approximately 88%. In comparison, the distribution of AVP-CSF-CN was mainly restricted to the rostral part of the third ventricle and the ratio of nNOS/AVP co-expression was only about 6%. The widespread presence of nNOS-CSF-CN-expressing OT in the third ventricular region suggests that NO is an important messenger in the CSF-hypothalamo-hypophyseal neuroendocrine regulation that may in part act in concert with OT. PMID- 15757632 TI - Asymmetrical changes of dopamine receptors in the striatum after unilateral dopamine depletion. AB - Dopamine plays an important role in modulating synaptic transmission in the striatum and has great influence on the function of the basal ganglia. Degeneration of dopamine neurons in the substantia nigra (SN) is the major cause of many neurological disorders, and the reduction of dopamine innervation results in alterations of dopamine receptors in the striatum. It has been shown that the nigrostriatal dopamine system has functional and neurochemical asymmetry. To investigate the lateralization of dopamine receptors in the striatum after dopamine denervation, the present study used quantitative autoradiography to compare the changes in dopamine receptor binding in the left and right striatum in rats after unilateral dopamine depletion. In comparison to control levels, dopamine D1)-like receptor binding, labeled with [3H]-SCH23390, in the dorsal striatum was reduced 2 weeks after unilateral lesions of the SN with 6 hydroxydopamine. D1-like receptor binding was decreased in the ipsilateral striatum following unilateral lesions of either the left or right SN. The left and right striatum responded similarly to unilateral SN lesions, as there were no significant differences in the percent decrease in D1-like binding in the two striata. In contrast, D2-like receptor binding, labeled with [3H]-spiroperidol, was significantly increased in the dorsal striatum following an ipsilateral SN lesion. Furthermore, the up-regulation of D2-like receptors in the right striatum was significantly greater than that in the left striatum after an ipsilateral lesion. The asymmetrical up-regulation of striatal D2 receptors after extensive dopamine depletion might contribute to the lateralization of the nigrostriatal system observed in some pathological conditions. PMID- 15757633 TI - Central administration of transforming growth factor-alpha and neuregulin-1 suppress active behaviors and cause weight loss in hamsters. AB - Transforming growth factor-alpha (TGF-alpha) is a candidate output signal of the hypothalamic circadian pacemaker. TGF-alpha is expressed in the suprachiasmatic nucleus (SCN) of rats, hamsters, and rhesus macaques [A. Kramer, F.C. Yang, P. Snodgrass, X. Li, T.E. Scammell, F.C. Davis and C.J. Weitz, Regulation of daily locomotor activity and sleep by hypothalamic EGF receptor signaling, Science, 294 (2001) 2511-5., X. Li, N. Sankrithi and F.C. Davis, Transforming growth factor alpha is expressed in astrocytes of the suprachiasmatic nucleus in hamster: role of glial cells in circadian clocks, Neuroreport, 13 (2002) 2143-7., Y.J. Ma, M.E. Costa and S.R. Ojeda, Developmental expression of the genes encoding transforming growth factor alpha and its receptor in the hypothalamus of female rhesus macaques, Neuroendocrinology, 60 (1994) 346-59., Y.J. Ma, M.P. Junier, M.E. Costa and S.R. Ojeda, Transforming growth factor-alpha gene expression in the hypothalamus is developmentally regulated and linked to sexual maturation, Neuron, 9 (1992) 657-70.]. TGF-alpha reversibly inhibits wheel-running activity during long-term infusions into the third ventricle of hamsters (2 weeks, intracerebroventricular or ICV) [A. Kramer, F.C. Yang, P. Snodgrass, X. Li, T.E. Scammell, F.C. Davis and C.J. Weitz, Regulation of daily locomotor activity and sleep by hypothalamic EGF receptor signaling, Science, 294 (2001) 2511-5.], and this effect appears to be mediated by the epidermal growth factor receptor (EGFR or ErbB-1) [A. Kramer, F.C. Yang, P. Snodgrass, X. Li, T.E. Scammell, F.C. Davis and C.J. Weitz, Regulation of daily locomotor activity and sleep by hypothalamic EGF receptor signaling, Science, 294 (2001) 2511-5.]. Here, we demonstrate that this inhibitory effect is not restricted to wheel-running behavior or to mediation by the EGFR. Using direct observation, we found the effects of long term TGF-alpha infusion (ICV, 12 microl/day, 3.3 microM) to be more general than previously reported. Other active behaviors such as grooming and feeding were reversibly inhibited and hamsters showed dramatic weight loss as a result of reduced feeding (34% of body weight over 19 days). TGF-alpha did not disrupt a non-behavioral rhythm, the rhythm in pineal melatonin. Wheel-running activity was also inhibited by another epidermal growth factor-like (EGF-like) peptide, neuregulin (NRG-1), that binds to different ErbB receptors. Like TGF-alpha, NRG-1 caused a significant weight loss. We also show that an acute injection of TGF alpha inhibits activity (ICV, 5 microl, 3.3 microM over 2 min), with inhibition and recovery occurring over a few hours. Although the results are consistent with the proposed [A. Kramer, F.C. Yang, P. Snodgrass, X. Li, T.E. Scammell, F.C. Davis and C.J. Weitz, Regulation of daily locomotor activity and sleep by hypothalamic EGF receptor signaling, Science, 294 (2001) 2511-5.] role for EGF like peptides in the daily regulation of activity, the actions of these peptides might also contribute to the behavioral etiology of diseases in which EGF-like peptides are expressed. PMID- 15757634 TI - VOR and Fos response during acute vestibular compensation in the Mongolian gerbil in darkness and in light. AB - We measured binocular horizontal eye movements in the gerbil following unilateral labyrinthectomy during the acute phase (1-24 h) of vestibular compensation. Regardless of whether the animals compensated in the light or the dark, VOR gain progressively reduced following the lesion, and normal oculomotor symmetry was disrupted. Initially, the VOR was comparable at 1 h post-lesion for both visual conditions. However, by 3 h post-lesion the VOR response for head turns away from the lesion continued to drop in animals compensating in the dark. By 24 h, both groups displayed reduced VOR gains, but animals compensating in the light had improved frequency response characteristics. Optokinetic responses became unstable but were generally elevated compared to pre-lesion levels. Animals with vision had reduced optokinetic gains by 24 h, while the OKR response for animals in the dark remained elevated. Brainstem Fos labeling generally increased from 1 to 3 h, then decreased by 24 h. However, at 1 h, Fos labeling in the inferior olivary dorsal cap and prepositus contralateral to the lesion was significantly increased in animals compensating in the light. In both visual conditions, flocculus and paraflocculus Purkinje cell labeling was also observed, and some of the Fos-labeled cells in the medial vestibular nucleus were commissural. Fos in the dorsal cap and prepositus could be attributed to the presence of visual input. While the visually related prepositus Fos labeling preceded improved VOR performance, the dorsal cap appeared to be involved in resolving visual and motor deficits from spontaneous nystagmus. PMID- 15757635 TI - Acute gastric changes after intracerebral hemorrhage in rats. AB - Severe intracerebral hemorrhage (ICH) produces gastric pathology in about 30% of the patient population, even after the standard treatment of H2 receptor blockers or proton pump inhibitors. This study was undertaken to establish a rat model of ICH-induced gastric ulcer. Adult male Sprague-Dawley rats (300-350 g) were divided into two hemorrhage groups and a sham control group. ICH was produced either by injection of 100 microl of autologous arterial blood or by injection of 4 microl saline containing 0.6 unit of bacterial collagenase VII into the right basal ganglia. Rats were sacrificed at 24, 48, 72 h, and 7 days after ICH to harvest brains and stomachs. Greater degrees of hemorrhage and brain edema were observed in collagenase-induced ICH. Motor behavior decreased significantly after 24 h in both models. The incidence of acute ulceration with destruction of the forestomach epithelium was extremely low at 8.7% in the collagenase injection model and 4.8% in the blood injection rats. Small, pinpoint hemorrhages (petechiae) were noticed in 38% of rats after blood injection and 22% after collagenase injection, in the glandular portion of the gastric mucosa with penetration of red blood cells and inflammatory cells into the gastric mucosa. Enhanced tumor necrosis factor alpha (TNFalpha) and cyclooxygenase 2 (COX-2) expressions were observed in gastric tissues after ICH with more intense staining occurring at 24 and 48 h. Due to the low incidence of ulceration, ICH-induced gastric ulceration in rodents may not appropriate for evaluating the potential human risk of gastric ulceration after ICH. PMID- 15757636 TI - Brain pericytes contribute to the induction and up-regulation of blood-brain barrier functions through transforming growth factor-beta production. AB - The blood-brain barrier (BBB) is a highly organized multicellular complex consisting of an endothelium, brain pericytes and astrocytes. The present study was aimed at evaluating the role of brain pericytes in the induction and maintenance of BBB functions and involvement of transforming growth factor-beta (TGF-beta) in the functional properties of pericytes. We used an in vitro BBB model established by coculturing immortalized mouse brain capillary endothelial (MBEC4) cells with a primary culture of rat brain pericytes. The coculture with rat pericytes significantly decreased the permeability to sodium fluorescein and the accumulation of rhodamine 123 in MBEC4 cells, suggesting that brain pericytes induce and up-regulate the BBB functions. Rat brain pericytes expressed TGF-beta1 mRNA. The pericyte-induced enhancement of BBB functions was significantly inhibited when cells were treated with anti-TGF-beta1 antibody (10 microg/ml) or a TGF-beta type I receptor antagonist (SB431542) (10 microM) for 12 h. In MBEC4 monolayers, a 12 h exposure to TGF-beta1 (1 ng/ml) significantly facilitated the BBB functions, this facilitation being blocked by SB431542. These findings suggest that brain pericytes contribute to the up-regulation of BBB functions through continuous TGF-beta production. PMID- 15757637 TI - Neuroprotective effects of an estratriene analog are estrogen receptor independent in vitro and in vivo. AB - Estrogens are potent neuroprotectants both in vitro and in vivo. In the present study, we compared the potency and efficacy of a non-feminizing estrogen, 2-(1 adamantyl)-4-methylestrone (ZYC-26), with its parent estrogen, estrone, and an expected non-neuroprotective 3-O-methyl analog of (17beta)-2-(1 adamantyl)estradiol (ZYC-23). These estratriene derivatives were tested for their ability to protect in an in vitro lipid peroxidation model, to neuroprotect against oxidative stress in cell culture models, to bind the estrogen receptors (ERalpha and ERbeta), to elicit uterotrophic effects, and to affect brain damage from transient middle cerebral artery occlusion. We observed that in contrast to estrone, neither ZYC-26 nor ZYC-23 bound to either estrogen receptors (ER) and both failed to elicit a uterotrophic response. In vitro, the active estrogen analogue ZYC-26 was more potent that estrogen in its ability to inhibit lipid peroxidation and to protect HT-22 cells from either glutamate or iodoacetic acid (IAA) toxicity. Further, ZYC-26 was as active in preventing brain damage from transient middle cerebral artery occlusion (MCAO) as was estrone. Collectively, these studies suggest that the antioxidant activity, rather than ER binding of non-feminizing estrogens such as ZYC-26, mediates their potent neuroprotective activity. Further, in view of the now known toxicities of chronic feminizing estrogen use in older women, non-feminizing estrogens may be a useful alternative for estrogen-induced brain protection. PMID- 15757638 TI - ERK1/2 are involved in low potassium-induced apoptotic signaling downstream of ASK1-p38 MAPK pathway in cultured cerebellar granule neurons. AB - We have recently reported that the ASK1-p38 MAPK pathway has an important role in the low potassium (LK)-induced apoptosis of cultured cerebellar granule neurons. In the present study, we observed that ERK1/2 were significantly activated 6 h after a change of medium from HK (high potassium) to LK. In addition, U0126, a specific inhibitor of MEKs, remarkably prevented the apoptosis of cultured cerebellar granule neurons. Then, we examined the mechanism underlying the activation of ERK1/2 in the LK-induced apoptotic pathway. The addition of SB203580, an inhibitor of p38 MAPK, suppressed the increase in the phosphorylation of ERK1/2 after the change to LK medium. Furthermore, we found that the expression of a constitutively active mutant of ASK1, an upstream kinase of p38 MAPK, enhanced the phosphorylation of ERK1/2. These results suggest that ERK1/2 play a crucial role in LK-induced apoptosis of cultured cerebellar granule neurons and that the LK-stimulated activation of ERK1/2 is regulated by the ASK1 p38 MAPK pathway. PMID- 15757639 TI - Cyclooxygenase inhibition attenuates endotoxin-induced spatial learning deficits, but not an endotoxin-induced blockade of long-term potentiation. AB - Peripheral administration of lipopolysaccharide (LPS), a potent bacterial endotoxin, can cause a variety of central effects, including production of cytokines and cyclooxygenases in the brain, as well as peripheral increases in corticosterone. These, in turn, may contribute to neuroimmune-induced neurocognitive deficits. We show here LPS causes deficits in hippocampal dependent spatial learning in the water maze but that treatment with ibuprofen, a broad-spectrum cyclooxygenase inhibitor, reverses the deficits induced in spatial learning by LPS. We also show that LPS causes an impairment in the induction of long-term potentiation in the dentate gyrus in vivo, a major contemporary model of learning and memory. No differences were found in corticosterone levels in trunk blood but we find a decrease in brain-derived neurotrophic factor (BDNF) expression in LPS group compared to saline controls. Paradoxically compared to the behavioral findings treatment with ibuprofen does not attenuate the LPS induced impairment in LTP or BDNF concentration in tetanized tissue. PMID- 15757640 TI - Spinal preprodynorphin mRNA expression in neonatal rats following peripheral inflammation. AB - Spinal nociceptive neural circuits undergo considerable changes during the postnatal period. This study showed that neonatal rats exhibited earlier upregulation and faster recovery of spinal preprodynorphin (PPD) mRNA than did the adults during complete Freund's adjuvant (CFA)-induced peripheral inflammation. These data suggest that the central nervous systems of neonates and adults respond differently to peripheral noxious inputs, a fact that should be considered when selecting pain treatment strategies for neonate populations. PMID- 15757641 TI - Single neuron activity changes to interleukin-1beta in the orbitofrontal cortex of the rat. AB - The orbitofrontal cortex (OBF) is known to play important roles in various regulatory processes. Our preliminary behavioral studies showed homeostatic alterations after orbitofrontal cortical microinjection of interleukin-1beta (IL 1beta) in the rat. To elucidate whether the above alterations were due to direct neuronal action of the cytokine, extracellular single neuron activity was recorded in the OBF of anesthetized rats by means of tungsten fiber multibarreled glass microelectrodes during microelectrophoretic administration of IL-1beta. More than half (56%) of all cells tested changed in firing rate in response to IL 1beta. Approximately 90% of these cytokine-modulated neurons were also sensitive to microelectrophoretically applied d-glucose, that is, proved to be the elements of the central glucose-monitoring neural network. The present findings demonstrate that IL-1beta can exert direct modulatory role on neurons in the OBF. PMID- 15757642 TI - Enhanced plasminogen activation by staphylokinase in the presence of streptokinase beta/betagamma domains: plasminogen kringles play a role. AB - Presence of isolated beta or betagamma domains of streptokinase (SK) increased the catalytic activity of staphylokinase (SAK)-plasmin (Pm) complex up to 60%. In contrast, fusion of SK beta or betagamma domains with the C-terminal end of SAK drastically reduced the catalytic activity of the activator complex. The enhancement effect mediated by beta or betagamma domain on Pg activator activity of SAK-Pm complex was reduced greatly (45%) in the presence of isolated kringles of Pg, whereas, kringles did not change cofactor activity of SAK fusion proteins (carrying beta or betagamma domains) significantly. When catalytic activity of SAK-microPm (catalytic domain of Pm lacking kringle domains) complex was examined in the presence of isolated beta and betagamma domains, no enhancement effect on Pg activation was observed, whereas, enzyme complex formed between microplasmin and SAK fusion proteins (SAKbeta and SAKbetagamma) displayed 50-70% reduction in their catalytic activity. The present study, thus, suggests that the exogenously present beta and betagamma interact with Pg/Pm via kringle domains and elevate catalytic activity of SAK-Pm activator complex resulting in enhanced substrate Pg activation. Fusion of beta or betagamma domains with SAK might alter these intermolecular interactions resulting in attenuated functional activity of SAK. PMID- 15757643 TI - Identification of EhICP1, a chagasin-like cysteine protease inhibitor of Entamoeba histolytica. AB - Based on the Entamoeba histolytica genome project (www.sanger.ac.uk/Project/E_histolytical/) we have identified a cysteine protease inhibitor, EhICP1 (amoebiasin 1), with significant homology to chagasin. Recombinant EhICP1 inhibited the protease activity of papain and that of a trophozoite lysate with Ki's in the picomolar range. By immunocytology, we localized the endogenous approximately 13 kDa EhICP1 in a finely dotted subcellular distribution discrete from the vesicles containing the amoebic cysteine protease, EhCP1 (amoebapain). In an overlay assay, we observed binding of recombinant EhICP1 to EhCP1. As a heptapeptide (GNPTTGF) corresponding to the second conserved chagasin motif inhibited the protease activity of both papain (K) 1.5 microM) and trophozoite extract (Ki in sub-mM range), it may be a candidate for the rational development of anti-amoebiasis drugs. PMID- 15757644 TI - The novel angiotensin II type 1 receptor (AT1R)-associated protein ATRAP downregulates AT1R and ameliorates cardiomyocyte hypertrophy. AB - Activation of angiotensin II (Ang II) type 1 receptor (AT1R) signaling is reported to play an important role in cardiac hypertrophy. We previously cloned a novel molecule interacting with the AT1R, which we named ATRAP (for Ang II type 1 receptor-associated protein). Here, we report that overexpression of ATRAP significantly decreases the number of AT1R on the surface of cardiomyocytes, and also decreases the degree of p38 mitogen-activated protein kinase phosphorylation, the activity of the c-fos promoter and protein synthesis upon Ang II treatment. These results indicate that ATRAP significantly promotes downregulation of the AT1R and further attenuates certain Ang II-mediated hypertrophic responses in cardiomyocytes. PMID- 15757645 TI - Downregulated mRNA expression of ASPP and the hypermethylation of the 5' untranslated region in cancer cell lines retaining wild-type p53. AB - The p53 protein is one of the best-known tumour suppressors. Recently discovered ASPP1 and ASPP2 are specific activators of p53. To understand, if apoptosis stimulating protein of p53 (ASPP) inactivation offers a selective advantage to tumors that have wild-type p53, we measured the mRNA expression of ASPP1 and ASPP2 in tumor cell lines retaining wide-type p53. In addition, the CpG island methylation status of ASPP1 gene and ASPP2 gene in the 5'-untranslated region was also investigated in order to understand the possible cause of abnormal expression of ASPP1 and ASPP2 in the tumor cell lines retaining wide-type p53. The data showed that mRNA expression of ASPP1 and ASPP2 is downregulated and CpG island tested is hypermethylated. These results indicated that ASPP CpG island aberrant methylation could be one molecular and genetic alteration in wild-type p53 tumours. PMID- 15757646 TI - A reassessment of the inhibitory capacity of human FKBP38 on calcineurin. AB - The microbial peptidomacrolide FK506 affects many eukaryotic developmental and cell signaling programs via calcineurin inhibition. Prior formation of a complex between FK506 and intracellular FK506-binding proteins (FKBPs) is the precondition for the interaction with calcineurin. A puzzling difference has emerged between the mammalian multidomain protein hFKBP38 and other FKBPs. It was shown that hFKBP38 not only binds to calcineurin but also inhibits the protein phosphatase activity of calcineurin on its own [Shirane, M. and Nakayama, K.I. (2003) Nature Cell Biol. 5, 28-37]. Inherent calcineurin inhibition by hFKBP38 would completely eliminate the need for FK506 in controlling many signal transduction pathways. To address this issue, we have characterized the functional and physical interactions between calcineurin and hFKBP38. A recombinant hFKBP38 variant and endogenous hFKBP38 were tested both in vitro and in vivo. The proteins neither directly inhibited calcineurin activity nor affected NFAT reporter gene activity in SH-SY5Y and Jurkat cells. In addition, a direct physical interaction between calcineurin and hFKBP38 was not detected in co-immunoprecipitation experiments. However, hFKBP38 indirectly affected the subcellular distribution of calcineurin by interaction with typical calcineurin ligands, as exemplified by the anti-apoptotic protein Bcl-2. Our data suggest that hFKBP38 cannot substitute for the FKBP/FK506 complex in signaling pathways controlled by the protein phosphatase activity of calcineurin. PMID- 15757647 TI - Inhibition of the angiogenesis by the MCP-1 (monocyte chemoattractant protein-1) binding peptide. AB - The CC chemokine, monocyte chemoattractant protein-1 (MCP-1), plays a crucial role in the initiation of atherosclerosis and has direct effects that promote angiogenesis. To develop a specific inhibitor for MCP-1-induced angiogenesis, we performed in vitro selection employing phage display random peptide libraries. Most of the selected peptides were found to be homologous to the second extracellular loops of CCR2 and CCR3. We synthesized the peptide encoding the homologous sequences of the receptors and tested its effect on the MCP-1 induced angiogenesis. Surface plasmon resonance measurements demonstrated specific binding of the peptide to MCP-1 but not to the other homologous protein, MCP-3. Flow cytometry revealed that the peptide inhibited the MCP-1 binding to THP-1 monocytes. Moreover, CAM and rat aortic ring assays showed that the peptide inhibited MCP-1 induced angiogenesis. Our observations indicate that the MCP-1 binding peptide exerts its anti-angiogenic effect by interfering with the interaction between MCP-1 and its receptor. PMID- 15757649 TI - The effect of calcium ions on subcellular localization of aldolase-FBPase complex in skeletal muscle. AB - In skeletal muscles, FBPase-aldolase complex is located on alpha-actinin of the Z line. In the present paper, we show evidence that stability of the complex is regulated by calcium ions. Real time interaction analysis, confocal microscopy and the protein exchange method have revealed that elevated calcium concentration decreases association constant of FBPase-aldolase and FBPase-alpha-actinin complex, causes fast dissociation of FBPase from the Z-line and slow accumulation of aldolase within the I-band and M-line. Therefore, the release of Ca2+ during muscle contraction might result, simultaneously, in the inhibition of glyconeogenesis and in the acceleration of glycolysis. PMID- 15757648 TI - Docking and fusion of insulin secretory granules in SUR1 knock out mouse beta cells observed by total internal reflection fluorescence microscopy. AB - To explore how the sulfonylurea receptor (SUR1) is involved in docking and fusion of insulin granules, dynamic motion of single insulin secretory granules near the plasma membrane was examined in SUR1 knock-out (Sur1KO) beta-cells by total internal reflection fluorescence microscopy. Sur1KO beta-cells exhibited a marked reduction in the number of fusion events from previously docked granules. However, the number of docked granules declined during stimulation as a consequence of the release of docked granules into the cytoplasm vs. fusion with the plasma membrane. Thus, the impaired docking and fusion results in decreased insulin exocytosis from Sur1KO beta-cells. PMID- 15757650 TI - Red and far-red light alter the transcript profile in the cyanobacterium Synechocystis sp. PCC 6803: impact of cyanobacterial phytochromes. AB - Cyanobacteria possess genes encoding phytochrome-related proteins. We used a DNA microarray approach to evaluate the impact of the phytochromes Cph1 and Cph2 on red light (R)- and far-red light (FR)-dependent gene expression in the unicellular cyanobacterium Synechocystis sp. PCC 6803. In cells of wild-type and phytochrome mutants, one-fourth of all 3165 annotated putative protein encoding genes was light-responsive. R predominantly enhanced the expression of genes involved in transcription, translation, and photosynthesis, whereas FR upregulated the transcript level of genes known to be inducible by stress. The absence of Cph1 and/or Cph2 altered the light-dependent expression of about 20 genes. Hence, receptor(s) different from the two phytochromes are supposed to trigger the global R/FR alterations of the expression profile. PMID- 15757651 TI - Comparative single-turnover kinetic analyses of trans-cleaving hammerhead ribozymes with naturally derived non-conserved sequence motifs. AB - trans-Cleaving hammerhead ribozyme variants were generated with mimicked non conserved internal loop motifs derived from five structurally diverse natural cis cleaving ribozymes. Most modified trans-cleaving variants showed enhanced single turnover cleavage rates relative to minimal counterparts that lack tertiary interactions between internal loop motifs I and II, and relative to controls with sequence changes in loop I. The trans-cleaving ribozyme derived from the positive strand of peach latent mosaic viroid had the highest observed cleavage rate, suggesting a structurally optimized motif that facilitates rapid formation of the ribozyme catalytic center in a trans-reaction. PMID- 15757652 TI - Matrix Mg2+ regulates mitochondrial ATP-dependent potassium channel from heart. AB - Mitochondrial ATP-regulated potassium (mitoKATP) channels play an important role in cardioprotection. Single channel activity was measured after reconstitution of inner mitochondrial membranes from bovine myocardium into a planar lipid bilayer. After incorporation, the potassium channel was recorded with a mean conductance of 103+/-9 pS. The channel activity was inhibited by ATP/Mg and activated by GDP. Magnesium ions alone affected, in a dose dependent manner, both the channel conductance and the open probability. Magnesium ions regulated the mitoKATP channel only when added to the trans compartment. We conclude that Mg2+ regulates the cardiac mitoKATP channel from the matrix site by affecting both the channel conductance and gating. PMID- 15757653 TI - Influence of hydrophobic matching on association of model transmembrane fragments containing a minimised glycophorin A dimerisation motif. AB - The principles that govern the folding and packing of membrane proteins are still not completely understood. In the present work, we have revisited the glycophorin A (GpA) dimerisation motif that mediates transmembrane (TM) helix association, one of the best-suited models of membrane protein oligomerisation. By using artificial polyleucine TM segments we have demonstrated in this study that a pattern of only five amino acids (GVxxGVxxT) promotes specific dimerisation. Further, we have used this minimised GpA motif to assess the influence of hydrophobic matching on the TM helix packing process in detergent micelles and found that this factor modulates helix-helix association and/or dissociation between TM fragments. PMID- 15757654 TI - Thyroid-hormone effects on putative biochemical pathways involved in UCP3 activation in rat skeletal muscle mitochondria. AB - In vitro, uncoupling protein 3 (UCP3)-mediated uncoupling requires cofactors [e.g., superoxides, coenzyme Q (CoQ) and fatty acids (FA)] or their derivatives, but it is not yet clear whether or how such activators interact with each other under given physiological or pathophysiological conditions. Since triiodothyronine (T3) stimulates lipid metabolism, UCP3 expression and mitochondrial uncoupling, we examined its effects on some biochemical pathways that may underlie UCP3-mediated uncoupling. T3-treated rats (Hyper) showed increased mitochondrial lipid-oxidation rates, increased expression and activity of enzymes involved in lipid handling and increased mitochondrial superoxide production and CoQ levels. Despite the higher mitochondrial superoxide production in Hyper, euthyroid and hyperthyroid mitochondria showed no differences in proton conductance when FA were chelated by bovine serum albumin. However, mitochondria from Hyper showed a palmitoyl-carnitine-induced and GDP-inhibited increased proton-conductance in the presence of carboxyatractylate. We suggest that T3 stimulates the UCP3 activity in vivo by affecting the complex network of biochemical pathways underlying the UCP3 activation. PMID- 15757655 TI - Both oxidative and nitrosative stress are associated with muscle wasting in tumour-bearing rats. AB - Reactive oxygen and nitrogen species (ROS and RNS) have been proposed as mechanisms of cancer-induced cachexia. In this study, we assessed using Western blot analysis the levels of total protein carbonylation (2,4 dinitrophenylhydrazine assay), both malondialdehyde- (MDA-) and 2-hydroxy-4 nonenal- (HNE-) protein adducts, Mn-superoxide dismutase (Mn-SOD), catalase, heme oxygenase-1 (HO-1) and 3-nitrotyrosine formation in gastrocnemius muscles of rats bearing the Yoshida AH-130 hepatoma. In the muscles of the tumour-bearing animals, protein carbonylation as measured by total levels of carbonyl group formation and both HNE and MDA-protein adducts, and protein tyrosine nitration were significantly greater than in control muscles. Protein levels of the antioxidant enzymes Mn-SOD, catalase, and HO-1 were not significantly modified in the rat cachectic muscles compared to controls. The inefficiency of the antioxidant enzymes in neutralizing excessive ROS production may account for elevated markers of protein oxidation and be responsible for the development of both oxidative and nitrosative stress in cancer-induced cachexia. PMID- 15757656 TI - Dietary polyphenols decrease glucose uptake by human intestinal Caco-2 cells. AB - The effect of different classes of dietary polyphenols on intestinal glucose uptake was investigated using polarised Caco-2 intestinal cells. Glucose uptake into cells under sodium-dependent conditions was inhibited by flavonoid glycosides and non-glycosylated polyphenols whereas aglycones and phenolic acids were without effect. Under sodium-free conditions, aglycones and non-glycosylated polyphenols inhibited glucose uptake whereas glycosides and phenolic acids were ineffective. These data suggest that aglycones inhibit facilitated glucose uptake whereas glycosides inhibit the active transport of glucose. The non-glycosylated dietary polyphenols appear to exert their effects via steric hindrance, and (-) epigallochatechingallate, (-)-epichatechingallate and (-)-epigallochatechin are effective against both transporters. PMID- 15757658 TI - Tangeretin inhibits extracellular-signal-regulated kinase (ERK) phosphorylation. AB - Tangeretin is a methoxyflavone from citrus fruits, which inhibits growth of human mammary cancer cells and cytolysis by natural killer cells. Attempting to unravel the flavonoid's action mechanism, we found that it inhibited extracellular-signal regulated kinases 1/2 (ERK1/2) phosphorylation in a dose- and time-dependent way. In human T47D mammary cancer cells this inhibition was optimally observed after priming with estradiol. The spectrum of the intracellular signalling kinase inhibition was narrow and comparison of structural congeners showed that inhibition of ERK phosphorylation was not unique for tangeretin. Our data add tangeretin to the list of small kinase inhibitors with a restricted intracellular inhibition profile. PMID- 15757657 TI - Cross-presentation of a CMV pp65 epitope by human dendritic cells using bee venom PLA2 as a membrane-binding vector. AB - We have used bee venom phospholipase A2 as a vector to load human dendritic cells ex vivo with a major histocompatibility complex (MHC) class I-restricted epitope fused to its C-terminus. The fusion protein bound to human monocyte-derived dendritic cells and was internalized into early endosomes. In vitro immunization experiments showed that these dendritic cells were able to generate specific CD8 T cell lines against the epitope carried by the fusion protein. Cross presentation did not require proteasome, transporter associated with antigen processing, or endosome proteases, but required newly synthesized MHC molecules. Comparison of the antigen presentation pathway observed in this study to that followed by other toxins used as vectors is discussed. PMID- 15757659 TI - Lithium inhibits brown adipocyte differentiation. AB - Lithium impairs the appearance of the characteristic morphology of brown adipocytes and downregulates the expression of marker genes of brown adipocyte differentiation. These effects are dose-dependent and are more pronounced when exposure of preadipocytes to lithium is initiated at early stages of differentiation. Although lithium reduces the expression of genes common to both white and brown adipocytes [fatty acid binding protein aP2 (aP2/FABP) or peroxisome proliferating activated receptor gamma], genes expressed differentially in brown adipocytes, i.e., uncoupling protein 1, PPAR gamma coactivator-1alpha, and peroxisome proliferating activated receptor alpha, are particularly sensitive to lithium treatment-dependent downregulation. Brown adipocytes appear as preferential targets of the inhibitory action of lithium on adipocyte differentiation. PMID- 15757661 TI - Identification of non-specific lipid transfer protein-1 as a calmodulin-binding protein in Arabidopsis. AB - Although non-specific lipid transfer proteins (nsLTPs) are widely present in plants, their functions and regulations have not been fully understood. In this report, Arabidopsis nsLTP1 was cloned and expressed to investigate its binding to calmodulin (CaM). Gel overlay assays revealed that recombinant nsLTP1 bound to CaM in a calcium-independent manner. The association of nsLTP1 and CaM was corroborated using CaM-Sepharose beads to specifically isolate recombinant nsLTP1 from crude bacterial lysate. The CaM-binding site was mapped in nsLTP1 to the region of 69-80 amino acids. This region is highly conserved among plant nsLTPs, implicating that nsLTPs are a new family of CaM-binding proteins whose functions may be mediated by CaM signaling. PMID- 15757660 TI - Ca2+-dependent K+ channels from rat olfactory cilia characterized in planar lipid bilayers. AB - Olfactory cilia contain cyclic nucleotide-gated and Ca2+-dependent Cl- conductances that underlie excitatory chemotransduction, and a Ca2+-dependent K+ (KCa) conductance, apparently involved in inhibitory transduction. Previous single-channel patch-clamp studies on olfactory cilia revealed four different KCas, with different conductances and kinetics. Here, we further characterized these channels in planar bilayers, where blockers could be properly tested. All four ciliary KCas were observed: The 16 pS channel, K0.5,Ca=40 microM and apamin sensitive; the 30 and 50 pS channel, K0.5,Ca=59 microM, clotrimazole-sensitive and charybdotoxin-insensitive; the 60 pS channel, clotrimazole-sensitive and charybdotoxin-insensitive; and the 210 pS channel, K0.5,Ca=63 microM, blocked by charybdotoxin and iberiotoxin. The presence of the 16 and 210 pS channels was confirmed by immunoblotting. PMID- 15757662 TI - Amino acid residues 62 and 193 play the key role in regulating the synergism of substrate binding in oyster arginine kinase. AB - The purpose of this study is to clarify the amino acid residues responsible for the synergism in substrate binding of arginine kinase (AK), a key enzyme in invertebrate energy metabolism. AKs contain a pair of highly conserved amino acids (D62 and R193) that form an ion pair, and replacement of these residues can cause a pronounced loss of activity. Interestingly, in the oyster Crassostrea AK, these residues are replaced by an N and a K, respectively. Despite this replacement, the enzyme retains high activity and moderate synergism in substrate binding (Kd/Km=2.3). We replaced the N62 by G or D and the K193 by G or R in Crassostrea AK, and also constructed the double mutants of N62G/K193G and N62D/K193R. All of the mutants retained 50-90% of the wild-type activity. In N62G and N62D mutants, the Kmarg for arginine binding was comparable to that of wild type enzyme, but the Kdarg was increased 2-5-fold, resulting in a strong synergism (Kd/Km=4.9-11.3). On the other hand, in K193G and K193R mutants, the Kmarg was increased 4-fold, and synergism was lost almost completely (Kd/Km=1.0 1.4). The N62G/K193G double mutant showed similar characteristics to the K193G and K193R mutants. Another double mutant, N62D/K193R, similar to the amino acid pair in the wild-type enzyme, had characteristics similar to those of the wild type enzyme. These results indicate that the amino acid residues 62 and 193 play the key role in mediating the synergism in substrate binding of oyster arginine kinase. PMID- 15757663 TI - Binding of JAB1/CSN5 to MIF is mediated by the MPN domain but is independent of the JAMM motif. AB - Macrophage migration inhibitory factor (MIF) binds to c-Jun activation domain binding protein-1 (JAB1)/subunit 5 of COP9 signalosome (CSN5) and modulates cell signaling and the cell cycle through JAB1. The binding domain of JAB1 responsible for binding to MIF is unknown. We hypothesized that the conserved Mpr1p Pad1p N terminal (MPN) domain of JAB1 may mediate binding to MIF. In fact, yeast two hybrid (YTH) and in vitro translation/coimmunoprecipitation (CoIP) analysis showed that a core MPN domain, which did not cover the functional JAB1/MPN/Mov34 metalloenzyme (JAMM) deneddylase sequence, binds to MIF comparable to full-length JAB1. YTH and pull-down analysis in conjunction with nanobead affinity matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry demonstrated that MIF(50-65) and MPN are sufficient to mediate MIF-JAB1 interaction, respectively. Finally, endogenous CoIP of MIF-CSN6 complexes from mammalian cells demonstrated that MPN is responsible for MIF-JAB1 binding in vivo, and, as CSN6 does not contain a functional JAMM motif, confirmed that the interaction does not require JAMM. PMID- 15757664 TI - Ultra-high resolution imaging of DNA and nucleosomes using non-contact atomic force microscopy. AB - Visualisation of nano-scale biomolecules aids understanding and development in molecular biology and nanotechnology. Detailed structure of nucleosomes adsorbed to mica has been captured in the absence of chemical-anchoring techniques, demonstrating the usefulness of non-contact atomic force microscopy (NC-AFM) for ultra-high resolution biomolecular imaging. NC-AFM offers significant advantages in terms of resolution, speed and ease of sample preparation when compared to techniques such as cryo-electron microscopy and X-ray crystallography. In the absence of chemical modification, detailed structure of DNA deposited on a gold substrate was observed for the first time using NC-AFM, opening up possibilities for investigating the electrical properties of unmodified DNA. PMID- 15757665 TI - Involvement of CD14 in the inhibitory effects of dimethyl-alpha-cyclodextrin on lipopolysaccharide signaling in macrophages. AB - The potential use of alpha-cyclodextrin and its hydrophilic alpha-cyclodextrin derivatives (alpha-CyDs) as antagonists against lipopolysaccharide (LPS), which stimulates the nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production as well as nuclear factor-kappaB (NF-kappaB) activation in macrophages was examined. Of three alpha-CyDs used in the present study, 2,6-di-O-methyl alpha-CyD (DM-alpha-CyD) had greater inhibitory activity than did the other CyDs against NO and TNF-alpha production through an impairment of gene expression in macrophage cell lines and primary macrophages stimulated with LPS and lipid A in a concentration-dependent manner. Concomitantly, DM-alpha-CyD inhibited NF-kappaB translocation into nucleus. These inhibitory effects of DM-alpha-CyD could be attributed to the release of CD14 from lipid rafts caused by an efflux of phospholipids, but not cholesterol. These results suggest that DM-alpha-CyD may have promise as a potent and unique antagonist for excess activation of macrophages stimulated with LPS. PMID- 15757666 TI - Genomic organization, promoter characterization and roles of Sp1 and AP-2 in the basal transcription of mouse PDIP1 gene. AB - The mouse polymerase delta-interacting protein 1 gene, PDIP1, is mapped to chromosome 7F3 region, spans approximately 16.7kb, and is organized into six exons. The transcription start site (TSS) was determined to be G, corresponding to position of 162-bp upstream of the translation start codon. The promoter region was found to lack TATA box or CCAAT box, instead, a CpG island was detected surrounding TSS. The region from -162 to +114 is required for basal transcriptional regulation of mouse PDIP1 gene, contains two AP-2 and two Sp1 binding sites. The Sp1 site upstream of TSS activates, while the other Sp1 site and two AP-2 sites suppress the transcription activity of mouse PDIP1 gene. PMID- 15757667 TI - Analysis of the mKir2.1 channel activity in potassium influx defective Saccharomyces cerevisiae strains determined as changes in growth characteristics. AB - Potassium uptake defective Saccharomyces cerevisiae strains (Deltatrk1,2 and Deltatrk1,2 Deltatok1) were used for the phenotypic analysis of the mouse inward rectifying Kir2.1 channel by growth analysis. Functional expression of both, multi-copy plasmid and chromosomally expressed GFP-mKir2.1 fusion constructs complemented the potassium uptake deficient phenotype in a pHout dependent manner. Upon application of Hygromycin B to chromosomally mKir2.1 expressing cells, significantly lower toxin sensitivity (EC50 15.4 microM) compared to Deltatrk1,2 Deltatok1 cells (EC50 2.6 microM) was observed. Growth determination of mKir2.1 expressing strains upon application of Ag+, Cs+ and Ba2+ as known blockers of mKir2.1 channels revealed significantly decreased channel function. Cells with mKir2.1 were about double sensitive to AgNO3, 350-fold more sensitive to CsCl and 1500-fold more sensitive to BaCl2 in comparison to the respective controls indicating functional expression and correct pharmacology. PMID- 15757668 TI - Expression profiling of the estrogen responsive genes in response to phytoestrogens using a customized DNA microarray. AB - Here, we examined phytoestrogens, isoflavones (genistein, daidzein, glycitein, biochanin A and ipriflavone), flavones (chrysin, luteolin and apigenin), flavonols (kaempferol and quercetin), and a coumestan, a flavanone and a chalcone (coumestrol, naringenin and phloretin, respectively) by means of a DNA microarray assay. A total of 172 estrogen responsive genes were monitored with a customized DNA microarray and their expression profiles for the above phytoestrogens were compared with that for 17beta-estradiol (E2) using correlation coefficients, or R values, after a correlation analysis by linear regression. While R values indicate the similarity of the response by the genes, we also examined the genes by cluster analysis and by their specificity to phytoestrogens (specific to genistein, daidzein or glycitein) or gene functions. Several genes were selected from p53-related genes (CDKN1A, TP53I11 and CDC14), Akt2-related genes (PRKCD, BRCA1, TRIB3 and APPL), mitogen-activated protein kinase-related genes (RSK and SH3BP5), Ras superfamily genes (RAP1GA1, RHOC and ARHGDIA) and AP-1 family and related genes (RIP140, FOS, ATF3, JUN and FRA2). We further examined the extracts from two local crops of soy beans (Kuro-daizu or Mochi-daizu) by comparing the gene expression profiles with those of E2 or phytoestrogens as a first step in utilizing the expression profiles for various applications. PMID- 15757669 TI - Direct interaction of coenzyme M with the active-site Fe-S cluster of heterodisulfide reductase. AB - Heterodisulfide reductase (HDR) catalyzes the formation of coenzyme M (CoM-SH) and coenzyme B (CoB-SH) by the reversible reduction of the heterodisulfide, CoM-S S-CoB. This reaction recycles the two thiol coenzymes involved in the final step of microbial methanogenesis. Electron paramagnetic resonance (EPR) and variable temperature magnetic circular dichroism spectroscopic experiments on oxidized HDR incubated with CoM-SH revealed a S=1/2 [4Fe-4S]3) cluster, the EPR spectrum of which is broadened in the presence of CoM-33SH [Duin, E.C., Madadi-Kahkesh, S., Hedderich, R., Clay, M.D. and Johnson, M.K. (2002) Heterodisulfide reductase from Methanothermobacter marburgensis contains an active-site [4Fe-4S] cluster that is directly involved in mediating heterodisulfide reduction. FEBS Lett. 512, 263 268; Duin, E.C., Bauer, C., Jaun, B. and Hedderich, R. (2003) Coenzyme M binds to a [4Fe-4S] cluster in the active site of heterodisulfide reductase as deduced from EPR studies with the [33S]coenzyme M-treated enzyme. FEBS Lett. 538, 81-84]. These results provide indirect evidence that the disulfide binds to the iron sulfur cluster during reduction. We report here direct structural evidence for this interaction from Se X-ray absorption spectroscopic investigation of HDR treated with the selenium analog of coenzyme M (CoM-SeH). Se K edge extended X ray absorption fine structure confirms a direct interaction of the Se in CoM-SeH treated HDR with an Fe atom of the Fe-S cluster at an Fe-Se distance of 2.4A. PMID- 15757671 TI - JM4 is a four-transmembrane protein binding to the CCR5 receptor. AB - The CC chemokine receptor 5 (CCR5) is a major co-receptor for human immunodeficiency virus (HIV) and CCR5 mutants lacking the carboxy (C)-terminus interfere with HIV infection. Therefore, we analysed the C-terminus of CCR5 and here describe Jena-Muenchen 4 (JM4), a novel CCR5-interacting protein. JM4 is membrane-associated, co-precipitates with CCR5, and is ubiquitously expressed. It shares about 62% sequence similarity with JWA and glutamate transporter associated protein 3-18 (GTRAP3-18), a regulator of an amino acid transporter. JWA, like JM4, is a four-transmembrane protein, which binds to the CCR5 receptor. Furthermore, JM4, JWA, and GTRAP3-18 co-localise and heterodimerise indicating a functional relationship. JM4 co-localises with calnexin in the endoplasmic reticulum and with the mannose 6-phosphate receptor in the Golgi. JM4 and GTRAP3 18 harbor a Rab-acceptor motif, indicating a function in vesicle formation at the Golgi complex. In conclusion, we describe a CCR5-interacting protein, which is suggested to function in trafficking and membrane localisation of the receptor, possibly also other receptors or amino acid transporters. PMID- 15757670 TI - Dictyostelium discoideum requires an Alix/AIP1 homolog, DdAlix, for morphogenesis in alkaline environments. AB - Alix and its homologs are involved in various phenomena such as endosomal protein sorting and adaptation to stress conditions. In this study, we found that development of Dictyostelium discoideum Alix (DdAlix) deletion mutant (alx-) cells was impaired in alkaline pH environments. The fruiting body formation efficiency of alx- cells at pH 9.0 was significantly lower than that of wild-type cells (6.8+/-4.2% vs 93+/-6.3%). The alkaline-sensitive phenotype of alx- cells was rescued by addition of salt. The phenotype was rescued by exogenous expression of human Alix as well as DdAlix but not by that of either Saccharomyces cerevisiae Alix homolog Rim20 or Bro1. DdAlix may be, structurally and functionally, more related to human Alix than to yeast Rim20 and Bro1. PMID- 15757672 TI - Formation of ceramide-enriched domains in lipid particles enhances the binding of apolipoprotein E. AB - We investigated the interaction between apolipoprotein E (apoE) and ceramide (CER)-enriched domains on the particles, by using lipid emulsions containing sphingomyelin (SM) or CER as model particles of lipoproteins. The sphingomyelinase (SMase)-induced aggregation of emulsion particles was prevented by apoE. CER increased the amount of apoE bound to emulsion particles. The confocal images of CER-containing large emulsions with two fluorescent probes showed three-dimensional microdomains enriched in CER. SMase also induced the formation of CER-enriched domains. We propose apoE prefers to bind on CER enriched domains exposed on particle surface, and thus inhibits the aggregation or fusion of the particles. PMID- 15757673 TI - Inhibition of beta-oxidative respiration is a therapeutic window associated with the cancer chemo-preventive activity of PPARgamma agonists. AB - We demonstrate expression and coordinate induction of PPARgamma and lipogenic enzymes (HMG-CoA synthase, HMG-CoA reductase and fatty acid synthase) in a murine lung alveolar carcinoma cell line (Line 1) treated with the PPARgamma agonist troglitazone (TRO) [0-100 microM]. We postulate that TRO induces a shift in cellular energy metabolism towards fatty acid oxidation (beta-oxidative respiration). Accordingly, co-treatment with TRO [30 microM] and increasing concentrations of trimetazidine (TMZ) [0.1-3 mM], an inhibitor of beta-oxidation, results in a dose dependent decrease cellular ATP levels and a dose dependent induction of apoptosis. These findings, suggest that inhibition of beta-oxidative respiration is a therapeutic window associated with the cancer chemo-preventive activity of PPARgamma agonists. PMID- 15757674 TI - Accumulation of deleterious mutations in hybridogenetic organisms. PMID- 15757675 TI - Stochastic modeling of T cell receptor gamma gene rearrangement. AB - The mechanisms controlling the recombination process of the gamma genes that encode the gamma chain of the antigen receptor of the gammadelta T lymphocytes are unclear. Based on experimental data on the recombination status of the two major TCR gamma genes expressed in V(gamma)4+ and V(gamma)1+ thymocytes, we tested the plausibility of three possible rearrangement mechanisms: (1) a time window mechanism according to which the two chromosomes are accessible to the recombination machinery during a defined period of time; (2) a feedback mechanism in which recombination stops shortly after the first in-frame rearrangement event anywhere in both chromosomes; and (3) a feedback mechanism with asynchronous chromosome accessibility, in which there is a first period when only one chromosome is accessible for recombination, followed by a second period when both chromosomes are accessible; shortly after the first in-frame rearrangement event, during any of these two periods, recombination will definitely stop. We model the time window mechanism using a pure probabilistic approach and the two feedback mechanisms using a continuous-time Markov chain formalism. We used maximum likelihood methodology to infer the goodness-of-fit of the models showing evidence for the last model, which best fits the data. Further analysis of this model suggests an evolutionary tradeoff between allelic and isotypic exclusion and the probability that a precursor differentiates into a mature gammadelta T lymphocyte. PMID- 15757676 TI - Daisyworld inhabited with daisies incorporating a seed size/number trade-off: the mechanism of negative feedback on selection from a standpoint of the competition theory. AB - We reexamined a Daisyworld model from the traditional view of competition theory. Unlike the original model, white and black daisies in our model incorporate a seeding/germination trade-off against bare ground area without assuming the local temperature reward. As a result, the planetary temperature is automatically regulated by two species if the following conditions are met: (i) the species react equally to an environmental condition, but one can alter the environmental condition in the opposite direction to the other. (ii) that one of the two cannot have both a higher maximal growth rate (mu(max)) and lower half-saturation constant (K) than those of the other. In other words, a pair of phenotypes incorporates a trade-off between quality and number of seeds. We found that the homeostatic regulation can also be reconciled with the adaptive evolution of optimal temperature. The results of simulation imply that biotic environmental feedback can also be maintained when the emergence of polymorphisms (black and white daisies) is closely linked to such a trade-off. PMID- 15757677 TI - Dynamical systems basis of metamorphosis: diversity and plasticity of cellular states in reaction diffusion network. AB - Dynamics maintaining diversity of cell types in a multi-cellular system are studied in relation to the plasticity of cellular states. By adopting a simple theoretical framework for intra-cellular chemical reaction dynamics and considering the division and death of cells, the development of cells is studied. Cell differentiation process is found to occur through instability in transient dynamics and cell-cell interaction. In long-term behavior, extinction of multiple cells is repeated, which leads to itinerancy over successive quasi-stable multi cellular states consisting of different types of cells. By defining the plasticity of a cellular state, it is shown that the plasticity of cells decreases before the extinction of most cell types, from which diversity and plasticity are later recovered. In the following, a decrease of plasticity occurs again, leading to the next extinction. This cycle of diversification and extinction is repeated. Relevance of our results to development and evolution is briefly discussed. PMID- 15757678 TI - Directed molecular evolution by machine learning and the influence of nonlinear interactions. AB - Alternative search strategies for the directed evolution of proteins are presented and compared with each other. In particular, two different machine learning strategies based on partial least-squares regression are developed: the first contains only linear terms that represent a given residue's independent contribution to fitness, the second contains additional nonlinear terms to account for potential epistatic coupling between residues. The nonlinear modeling strategy is further divided into two types, one that contains all possible nonlinear terms and another that makes use of a genetic algorithm to select a subset of important interaction terms. The performance of each modeling type as a function of training set size is analysed. Simulated molecular evolution on a synthetic protein landscape shows the use of machine learning techniques to guide library design can be a powerful addition to library generation methods such as DNA shuffling. PMID- 15757679 TI - Finding optimal vaccination strategies for pandemic influenza using genetic algorithms. AB - In the event of pandemic influenza, only limited supplies of vaccine may be available. We use stochastic epidemic simulations, genetic algorithms (GA), and random mutation hill climbing (RMHC) to find optimal vaccine distributions to minimize the number of illnesses or deaths in the population, given limited quantities of vaccine. Due to the non-linearity, complexity and stochasticity of the epidemic process, it is not possible to solve for optimal vaccine distributions mathematically. However, we use GA and RMHC to find near optimal vaccine distributions. We model an influenza pandemic that has age-specific illness attack rates similar to the Asian pandemic in 1957-1958 caused by influenza A(H2N2), as well as a distribution similar to the Hong Kong pandemic in 1968-1969 caused by influenza A(H3N2). We find the optimal vaccine distributions given that the number of doses is limited over the range of 10-90% of the population. While GA and RMHC work well in finding optimal vaccine distributions, GA is significantly more efficient than RMHC. We show that the optimal vaccine distribution found by GA and RMHC is up to 84% more effective than random mass vaccination in the mid range of vaccine availability. GA is generalizable to the optimization of stochastic model parameters for other infectious diseases and population structures. PMID- 15757680 TI - Simulated evolution of selfish herd behavior. AB - Single species aggregations are a commonly observed phenomenon. One potential explanation for these aggregations is provided by the selfish herd hypothesis, which states that aggregations result from individual efforts to reduce personnel predation risk at the expense of group-mates. Not all movement rules based on the selfish herd hypothesis are consistent with observed animal behavior. Previous work has shown that herd-like aggregations are not generated by movement rules limited to local interactions between nearest neighbors. Instead, rules generating realistic herds appear to require delocalized interactions. To date, it has been an open question whether or not the necessary delocalization can emerge from local interactions under natural selection. To address this question, we study an individual-based model with a single quantitative genetic trait that controls the influence of neighbors as a function of distance. The results indicate that predation-based selection can increase the influence of distant neighbors relative to near neighbors. Our results lend support for the idea that selfish herd behavior can arise from localized movement rules under natural selection. PMID- 15757681 TI - Diversity in times of adversity: probabilistic strategies in microbial survival games. AB - Population diversification strategies are ubiquitous among microbes, encompassing random phase-variation (RPV) of pathogenic bacteria, viral latency as observed in some bacteriophage and HIV, and the non-genetic diversity of bacterial stress responses. Precise conditions under which these diversification strategies confer an advantage have not been well defined. We develop a model of population growth conditioned on dynamical environmental and cellular states. Transitions among cellular states, in turn, may be biased by possibly noisy readings of the environment from cellular sensors. For various types of environmental dynamics and cellular sensor capability, we apply game-theoretic analysis to derive the evolutionarily stable strategy (ESS) for an organism and determine when that strategy is diversification. We find that: (1) RPV, effecting a sort of Parrondo paradox wherein random alternations between losing strategies produce a winning strategy, is selected when transitions between different selective environments cannot be sensed, (2) optimal RPV cell switching rates are a function of environmental lifecycle asymmetries and environmental autocorrelation, (3) probabilistic diversification upon entering a new environment is selected when sensors can detect environmental transitions but have poor precision in identifying new environments, and (4) in the presence of excess additive noise, low-pass filtering is required for evolutionary stability. We show that even when RPV is not the ESS, it may minimize growth rate variance and the risk of extinction due to 'unlucky' environmental dynamics. PMID- 15757682 TI - A microbial modified prisoner's dilemma game: how frequency-dependent selection can lead to random phase variation. AB - Random phase variation (RPV) is a control strategy in which the expression of a cell state or phenotype randomly alternates between discrete 'on' and 'off' states. Though this mode of control is common for bacterial virulence factors like pili and toxins, precise conditions under which RPV confers an advantage have not been well defined. In Part I of this study, we predicted that fluctuating environments select for RPV if transitions between different selective environments cannot be reliably sensed (J. Theor. Biol. (2005)). However, selective forces both inside and outside of human hosts are also likely to be frequency dependent in the sense that the fitnesses of some bacterial states are greatest when rare. Here we show that RPV at slow rates can provide a survival advantage in such a frequency-dependent environment by generating population heterogeneity, essentially mimicking a polymorphism. More surprisingly, RPV at a faster 'optimal' rate can shift the population composition toward an optimal growth rate that exceeds that possible for polymorphic populations, but this optimal strategy is not evolutionarily stable. The population would be most fit if all cells randomly phase varied at the optimal rate, but individual cells have a growth-rate incentive to defect (mutate) to other switching rates or non-phase variable phenotype expression, leading to an overall loss of fitness of the individual and the population. This scenario describes a modified Prisoner's Dilemma game (Evolution and the Theory of Games, Cambridge University Press, Cambridge, New York, 1982, viii, 224pp.; Nature 398 (6726) (1999) 367), with random phase variation at optimal switching rates serving as the cooperation strategy. PMID- 15757683 TI - A model for length control of flagellar hooks of Salmonella typhimurium. AB - We present a mathematical model for the growth and length regulation of the hook component of the flagellar motor of Salmonella typhimurium. Under the assumption that the molecular constituents are translocated into the nascent filament by an ATP-ase and then move by molecular diffusion to the growing end, where they polymerize into the growing tube, we find that there is a detectable transition from secretion limited growth to diffusion limited growth. We propose that this transition can be detected by the secretant FliK, allowing FliK to interact with FlhB thereby changing the secretion target of the type III secretion machinery and terminating the growth of the hook. PMID- 15757684 TI - On the importance of fatty acid composition of membranes for aging. AB - The membrane pacemaker theory of aging is an extension of the oxidative stress theory of aging. It emphasises variation in the fatty acid composition of membranes as an important influence on lipid peroxidation and consequently on the rate of aging and determination of lifespan. The products of lipid peroxidation are reactive molecules and thus potent damagers of other cellular molecules. It is suggested that the feedback effects of these peroxidation products on the oxidative stress experienced by cells is an important part of the aging process. The large variation in the chemical susceptibility of individual fatty acids to peroxidation coupled with the known differences in membrane composition between species can explain the different lifespans of species, especially the difference between mammals and birds as well as the body-size-related variation in lifespan within mammals and birds. Lifespan extension by calorie-restriction can also be explained by changes in membrane fatty acid composition which result in membranes more resistant to peroxidation. It is suggested that lifespan extension by reduced insulin/IGF signalling may also be mediated by changes in membrane fatty acid composition. PMID- 15757685 TI - Mathematical modelling of prolactin-receptor interaction and the corollary for prolactin receptor gene expression in skin. AB - A mathematical model of prolactin regulating its own receptors was developed, and compared with experimental data on a qualitative level. The model incorporates the kinetics of prolactin-receptor interactions and subsequent signalling by prolactin-receptor dimers to regulate the production of receptor mRNA and hence the receptor population. The model relates changes in plasma prolactin concentration to prolactin receptor (PRLR) gene expression, and can be used for predictive purposes. The cell signalling that leads to the activation of target genes, and the mechanisms for regulation of transcription, were treated empirically in the model. The model's parameters were adjusted so that model simulations agreed with experimentally observed responses to administration of prolactin in sheep. In particular, the model correctly predicts insensitivity of receptor mRNA regulation to a series of subcutaneous injections of prolactin, versus sensitivity to prolonged infusion of prolactin. In the latter case, response was an acute down-regulation followed by a prolonged up-regulation of mRNA, with the magnitude of the up-regulation increasing with the duration of infusion period. The model demonstrates the feasibility of predicting the in vivo response of prolactin target genes to external manipulation of plasma prolactin, and could provide a useful tool for identifying optimal prolactin treatments for desirable outcomes. PMID- 15757686 TI - Lines in the Western Australian sand. PMID- 15757687 TI - Biodiversity reduction on contaminated sediments in the sea: contamination or sediment mobility? AB - Where contaminants in sediments correlate with biodiversity reductions, the contamination may not be the cause. Other physical factors may be responsible, and should be investigated so that any remedial action is directed properly. Two examples are given where unusual sediment mobility caused biodiversity reductions in sediments with elevated trace contaminants. PMID- 15757688 TI - Comparison of environmental fate and transport process descriptors of explosives in saline and freshwater systems. AB - Environmental process descriptors are necessary to evaluate the fate and transport of munitions constituents that have been introduced into the environment. An extensive database exists for freshwater environments; however, explosives fate and transport parameters such as dissolution rates, transformation rates, and adsorption of explosives have not been evaluated under both freshwater and saline conditions to determine the applicability of the freshwater data to saline environments. The objective of this study was to determine if freshwater fate and transport processes were similar to those determined under saline water conditions. We evaluated TNT, RDX, and HMX dissolution rates, transformation rates, and adsorption under freshwater and saline conditions in batch tests. Results showed a generally close agreement. Therefore, the existing freshwater database for explosives fate and transport process descriptors can be used in marine environments. PMID- 15757689 TI - Identification and quantification of pesticides, industrial chemicals, and organobromine compounds of medium to high polarity in the North Sea. AB - Solid-phase extraction of 20 L seawater samples enabled the enrichment and determination of a wide array of organic substances, including compounds of medium to high polarity, in the pg/L-range. A number of contaminants was detected and quantified throughout the North Sea, among them the pesticides dichlobenil (2,6-dichlorobenzonitrile), metolachlor and terbuthylazine as well as the industrial chemicals dichloropyridines (DCPy, 4 isomers) and nitrobenzene. Concentrations attained values up to 1.4 ng/L for dichlobenil, 0.83 ng/L for terbuthylazine, 0.61 ng/L for metolachlor, 0.13 ng/L for 2,6-DCPy, 4.37 ng/L for nitrobenzene and 1-8 ng/L for tris(chloropropyl)phosphates (TCPP). A number of North Sea water samples was screened for non-target compounds, revealing the presence of further contaminants, e.g., lindane and TCPP, as well as several biogenic and/or anthropogenic organobromine substances, among which bromoindols, phenols and -anisoles were identified. PMID- 15757690 TI - Benthic diatom community response to environmental variables and metal concentrations in a contaminated bay adjacent to Casey Station, Antarctica. AB - This study examined the effects of anthropogenic contaminants and environmental variables on the composition of benthic diatom communities within a contaminated bay adjacent to an abandoned waste disposal site in Antarctica. The combination of geographical, environmental and chemical data included in the study explained all of the variation observed within the diatom communities. The chemical data, particularly metal concentrations, explained 45.9% of variation in the diatom communities, once the effects of grain-size and spatial structure had been excluded. Of the metals, tin explained the greatest proportion of variation in the diatom communities (28%). Tin was very highly correlated (R2>0.95) with several other variables (copper, iron, lead, and sum of metals), all of which explained similarly high proportions of total variation. Grain-size data explained 23% of variation once the effects of spatial structure and the chemical data had been excluded. The pure spatial component explained only 1.8% of the total variance. The study demonstrates that much of the compositional variability observed in the bay can be explained by concentrations of metal contaminants. PMID- 15757691 TI - Constraints on spatial variability in soft-sediment communities affected by contamination from an Antarctic waste disposal site. AB - A small-scale (<500 m length) transect-based survey was conducted in December 1998 to examine the spatial distribution of soft-sediment communities and of concentrations of heavy metals and hydrocarbons in sediments in Brown Bay, adjacent to an abandoned waste dump, at Casey Station, Antarctica. Samples were taken along three transects at increasing distances (nine stations) from the shore-line waste dump. A gradient of contamination was detected, but concentrations of contaminants were very variable with "hotspots" or high levels of contaminants at some stations. Multivariate analysis revealed that the distribution of soft-sediment communities was distinctly different between the inner, middle and outer stations. Abundances of most taxa were very variable with few patterns apparent, but some fauna displayed an abundance gradient from the inner to the outer part of the bay. Many taxa had maximum abundances at outer stations and minimum at inner stations. Multivariate correlations between environmental variables and soft-sediment communities indicated that combinations of some metals (Cd, Cu, Sn, Pb) and grain size (mainly finer fractions, fine sands and coarse silts) were the variables that best "matched" the community patterns within Brown Bay. This study indicated that there were significant correlations between the presence of contaminants and the distribution and composition of soft-sediment communities over very small spatial scales. PMID- 15757692 TI - Mercury concentrations in red drum, Sciaenops ocellatus, from estuarine and offshore waters of Florida. AB - Dorsal muscle tissue from 712 red drum, Sciaenops ocellatus, from Florida waters were analyzed for total mercury content. Mercury levels detected in these red drum varied but in most study areas were usually lower than regulatory threshold guidelines. Total mercury levels in individual fish from all study areas ranged from 0.020 to 3.6 ppm (wet weight). Total mercury levels detected in red drum from the Florida Keys-Florida Bay area were often higher than those in fish from all other estuarine study areas. Positive relationships between total mercury levels and fish size (length and weight) and fish age were observed in most Florida study areas, indicating that mercury levels tend to increase over time as red drum grow. The majority of large, mature red drum examined contained mercury levels greater than the 0.5-ppm threshold level set by the Florida Department of Health (DOH). Approximately 94% of all adult red drum from offshore waters adjacent to Tampa Bay contained mercury levels greater than or equal to the 0.5 ppm threshold level, and 64% contained levels greater than or equal to the DOH 1.5-ppm "no consumption" level. All fish from this area with mercury levels greater than 1.5 ppm were large individuals (670 mm SL). Eight percent of legal size red drum from Florida waters contained total mercury levels greater than or equal to the 0.5-ppm threshold level. The majority (52%) of these legal-size fish greater than or equal to 0.5 ppm were from the Florida Keys-Florida Bay area. In the Florida recreational fishery, the current maximum size limit for this species is an effective filter that prevents humans from consuming those red drum with the greatest likelihood of containing high mercury levels. PMID- 15757693 TI - Effects of 12 years' operation of a sewage treatment plant on trace metal occurrence within a Mediterranean commercial sponge (Spongia officinalis, Demospongiae). AB - The present field study uses Spongia officinalis for assessing trace metals occurrence in time and space within Mediterranean rocky communities. Nine sites were selected in the Marseille area for studying spatial trends in 12 metal concentrations. Long term changes in 8 metal concentrations were assessed at sites that had been sampled before and 12 years after the opening of a treatment plant. Spongia officinalis highly concentrated all the trace metal surveyed excepted Hg and Cd. The overall contamination level registered provided a classification of the study sites which is congruent with that given by other studies on pollutant accumulation in neighbouring sandy-bottoms or benthic assemblages. Among the metals studied, Fe, Pb, Cr are those that best highlighted a pollution gradient. In the present study, only Cd concentration did not vary in space. Except for Ni, all pollutant concentrations clearly decreased between 1984 and 1999. This very impressive decrease in heavy metal concentrations within the Marseille area represents an indisputable evidence of the improvement of the seawater quality resulting from 12 years' operation of the Marseille sewage plant. Moreover, the significant decrease also recorded in the reference population at Port-Cros might reflect an overall improvement in the seawater quality of the NW Mediterranean. PMID- 15757694 TI - The use of steroid markers to assess sewage contamination of the Black Sea. AB - Analyses of faecal steroids in coastal sediments taken from throughout the Black Sea indicate chronic sewage contamination at some locations. These include Sochi, where concentrations of coprostanol up to 5400 ng g(-1) (dry wt) were recorded, and in the coastal areas adjacent to the Danube delta (2600 ng g(-1) dry wt). Comparatively high values of 5beta/(5beta+5alpha) isomeric ratios of cholestan-3 ol and cholestan-3-one are reported at these locations and are characteristic of sediments contaminated with sewage. Lower concentrations of coprostanol are reported for Bosphorus sediments (12-440 ng g(-1) dry wt) and in the region of Odessa (130-290 ng g(-1) dry wt). Isomeric ratios at these locations also confirm sewage as a significant contributor to steroids in the Bosphorus, but lower values of the ratios indicate only minor sewage inputs in Odessa. In contrast, steroid concentrations and compositions from most Ukrainian sites are characteristic of uncontaminated environments. To place these results into perspective, the range in levels of coprostanol in the Black Sea sediments compares to the lower to mid-range of concentrations reported for coastal sediments on a world-wide basis, with Black Sea sedimentary levels substantially below those previously reported for heavily impacted sites. PMID- 15757695 TI - Spatio-temporal distribution and characteristics of PAHs in sediments from Masan Bay, Korea. AB - The distribution of polycyclic aromatic hydrocarbons (PAHs) has been investigated in the surface and core sediments from Masan Bay, Korea. Total PAHs in the surface sediments ranged from 207 to 2670 ng/g dry weight with a mean value of 680 ng/g. Qualitative similarity and quantitative difference between inner and outer bay indicate that the main sources of PAHs are located in the inner bay and outer bay is also affected by the same sources. Vertical distribution of PAHs revealed that three distinctive stages could be differentiated with the help of PCA analysis. The highest concentration (industrialization stage) appeared between late 1950s and 1980, which was 10 years later than other developed countries. A strong pyrolytic source fingerprint has been detected with slight influence of petrogenic sources, and diagenetic PAH, perylene also contributed. Total organic carbon normalized PAHs (sum of 13 PAHs, 8.85-88.0 microg/g OC) were under the threshold effects concentration (TEC, 290 microg/g OC). PMID- 15757696 TI - Phytoplankton distributions and their relationship with the environment in the Changjiang Estuary, China. AB - This study was carried out in the Changjiang Estuary from 19 to 26 May 2003. Based on the data collected from 29 stations, including two anchor stations, phytoplankton taxonomic composition, abundance, diurnal variability and spatial distribution were examined. Eighty-seven species, including 54 species of diatoms and 16 red tide causative species, were identified. Average diversity index (H') and evenness (J) values were 1.04 and 0.40, respectively. A bloom in abundance of certain phytoplankton species, especially Prorocentrum dentatum and Skeletonema costatum, was thought to be the cause of the low diversity index and evenness values. Total phytoplankton abundance averaged 6.75 x 10(5)cells l(-1), and was much higher than previous investigation carried out in the same month in 1986. Abundance increased seaward showing a distinct spatial difference, and the dominant species varied with salinity. Correlation between phosphorus and abundance further supported the former conclusion that phosphorus is the controlling factor in phytoplankton growth in the Changjiang Estuary where light is not limiting. Based on the relationship between DO, pH and abundance, it is likely that the bloom was caused by rapid in situ growth of phytoplankton with high nutrients and sufficient light. The data also indicated that the duration of the bloom was not long and 2 microm) of the inshore central GBR shelf in the wet season when A. planci larvae develop, is double that of other places and times. Larval development, growth and survival increase almost ten-fold with doubled concentrations of large phyto-plankton. This and other lines of evidence suggest that frequent A. planci outbreaks on the GBR may indeed be a result of increased nutrient delivery from the land. PMID- 15757728 TI - Water quality in the Great Barrier Reef region: responses of mangrove, seagrass and macroalgal communities. AB - Marine plants colonise several interconnected ecosystems in the Great Barrier Reef region including tidal wetlands, seagrass meadows and coral reefs. Water quality in some coastal areas is declining from human activities. Losses of mangrove and other tidal wetland communities are mostly the result of reclamation for coastal development of estuaries, e.g. for residential use, port infrastructure or marina development, and result in river bank destabilisation, deterioration of water clarity and loss of key coastal marine habitat. Coastal seagrass meadows are characterized by small ephemeral species. They are disturbed by increased turbidity after extreme flood events, but generally recover. There is no evidence of an overall seagrass decline or expansion. High nutrient and substrate availability and low grazing pressure on nearshore reefs have lead to changed benthic communities with high macroalgal abundance. Conservation and management of GBR macrophytes and their ecosystems is hampered by scarce ecological knowledge across macrophyte community types. PMID- 15757729 TI - Effects of Photosystem II inhibiting herbicides on mangroves--preliminary toxicology trials. AB - Mangroves are sensitive to the root application of Photosystem II inhibiting herbicides and Avicennia marina is more sensitive than other mangroves tested. Seedlings of four mangrove species, including two salt-excreting species (A. marina and Aegiceras corniculatum) and two salt-excluding species (Rhizophora stylosa and Ceriops australis) were treated with a range of concentrations of the herbicides diuron, ametryn and atrazine. Assessment of responses required the separation of seedlings into two groups: those that had only their roots exposed to the herbicides through the water (A. marina and R. stylosa) and those that had both roots and leaves exposed to herbicides through the water (A. corniculatum and C. australis). Salt-excreting species in each group were more susceptible to all herbicide treatments than salt-excluding species, indicating that root physiology was a major factor in the uptake of toxic pollutants in mangroves. Submergence of leaves appeared to facilitate herbicide uptake, having serious implications for seedling recruitment in the field. Each herbicide was ranked by its toxicity to mangrove seedlings from most damaging to least effective, with diuron>ametryn>atrazine. The relative sensitivity of A. marina found in these pot trials was consistent with the observed sensitivity of this species in the field, notably where severe dieback had specifically affected A. marina in the Mackay region, north eastern Australia. PMID- 15757730 TI - Herbicides implicated as the cause of severe mangrove dieback in the Mackay region, NE Australia: consequences for marine plant habitats of the GBR World Heritage Area. AB - Herbicides, particularly diuron, were correlated with severe and widespread dieback of the dominant mangrove, Avicennia marina (Forsk.) Vierh. var. eucalyptifolia (Val.) N.C. Duke (Avicenniaceae), its reduced canopy condition, and declines in seedling health within three neighbouring estuaries in the Mackay region of NE Australia. This unusual species-specific dieback, first observed in the early 1990s, had gotten notably worse by 2002 to affect >30 km(2) of mangroves in at least five adjacent estuaries in the region. Over the past century, agricultural production has responded well to the demands of increasing population with improvements in farm efficiency assisted by significant increases in the use of agricultural chemicals. However, with regular and episodic river flow events, these chemicals have sometimes found their way into estuarine and nearshore water and sediments where their effects on marine habitats have been largely unquantified. Investigations over the last three years in the Mackay region provide compelling evidence of diuron, and possibly other agricultural herbicides, as the most likely cause of the severe and widespread mangrove dieback. The likely consequences of such dieback included declines in coastal water quality with increased turbidity, nutrients and sediment deposition, as well as further dispersal of the toxic chemicals. The implications of such findings are immense since they describe not only the serious deterioration of protected and beneficial mangrove habitat but also the potential for significant direct and indirect effects on other highly-valued estuarine and marine habitats in the region, including seagrass beds and coral reefs of the Great Barrier Reef lagoon. This article reviews all key findings and observations to date and describes the essential correlative and causative evidence. PMID- 15757731 TI - Herbicide contamination and the potential impact to seagrass meadows in Hervey Bay, Queensland, Australia. AB - Low concentrations of herbicides (up to 70 ng l(-1)), chiefly diuron (up to 50 ng l(-1)) were detected in surface waters associated with inter-tidal seagrass meadows of Zostera muelleri in Hervey Bay, south-east Queensland, Australia. Diuron and atrazine (up to 1.1 ng g(-1) dry weight of sediment) were detected in the sediments of these seagrass meadows. Concentration of the herbicides diuron, simazine and atrazine increased in surface waters associated with seagrass meadows during moderate river flow events indicating herbicides were washed from the catchment to the marine environment. Maximum herbicide concentration (sum of eight herbicides) in the Mary River during a moderate river flow event was 4260 ng l(-1). No photosynthetic stress was detected in seagrass in this study during low river flow. However, with moderate river flow events, nearshore seagrasses are at risk of being exposed to concentrations of herbicides that are known to inhibit photosynthesis. PMID- 15757732 TI - Variation in biogeochemical parameters across intertidal seagrass meadows in the central Great Barrier Reef region. AB - This survey provides baseline information on sediment characteristics, porewater, adsorbed and plant tissue nutrients from intertidal coastal seagrass meadows in the central region of the Great Barrier Reef World Heritage Area. Data collected from 11 locations, representative of intertidal coastal seagrass beds across the region, indicated that the chemical environment was typical of other tropical intertidal areas. Results using two different extraction methods highlight the need for caution when choosing an adsorbed phosphate extraction technique, as sediment type affects the analytical outcome. Comparison with published values indicates that the range of nutrient parameters measured is equivalent to those measured across tropical systems globally. However, the nutrient values in seagrass leaves and their molar ratios for Halophila ovalis and Halodule uninervis were much higher than the values from the literature from this and other regions, obtained using the same techniques, suggesting that these species act as nutrient sponges, in contrast with Zostera capricorni. The limited historical data from this region suggest that the nitrogen and phosphorus content of seagrass leaves has increased since the 1970s concomitant with changing land use practice. PMID- 15757733 TI - Seagrass population dynamics and water quality in the Great Barrier Reef region: a review and future research directions. AB - Seagrasses in the Great Barrier Reef region, particularly in coastal habitats, act as a buffer between catchment inputs and reef communities and are important habitat for fisheries and a food source for dugong and green turtle. Within the Great Barrier Reef region there are four different seagrass habitat types now recognised. The spatial and temporal dynamics of the different types of seagrass habitat is poorly understood. In general seagrass growth is limited by light, disturbance and nutrient supply, and changes to any or all of these limiting factors may cause seagrass decline. The capacity of seagrasses to recover requires either recruitment via seeds or through vegetative growth. The ability of seagrass meadows to recover from large scale loss of seagrass cover observed during major events such as cyclones or due to anthropogenic disturbances such as dredging will usually require regeneration from seed bank. Limited research into the role of pollutants on seagrass survival suggests there may be ongoing impacts due to herbicides, pesticides and other chemical contaminants. Further research and monitoring of seagrass meadow dynamics and the influence of changing water quality on these is needed to enhance our ability to manage seagrasses on the Great Barrier Reef. PMID- 15757734 TI - Application of a novel phytotoxicity assay for the detection of herbicides in Hervey Bay and the Great Sandy Straits. AB - A novel phytotoxicity assay was incorporated into an environmental assessment of Hervey Bay and the Great Sandy Straits, to investigate the role of run-off associated herbicides in the deteriorated health of intertidal seagrass meadows. Dose response curves of common herbicides were performed and their toxicity equivalents elucidated to assist in analysis. The results of the assay were reproducible and corresponded strongly with results of chemical analyses. The incorporation of the assay into the assessment of surface waters added an important aspect to the study by allowing investigation of the toxicity of cumulative herbicide concentrations and yielding biologically relevant data. The highest herbicide concentration detected during the study was equivalent to 0.23 microg l(-1) diuron; a concentration known to inhibit photosynthetic efficiency of the assay biomaterial by approximately 3%. PMID- 15757735 TI - Organochlorine and heavy metal concentrations in blubber and liver tissue collected from Queensland (Australia) dugong (Dugong dugon). AB - Tissue samples of liver and blubber were salvaged from fifty-three dugong (Dugong dugon) carcasses stranded along the Queensland coast between 1996 and 2000. Liver tissue was analysed for a range of heavy metals and blubber samples were analysed for organochlorine compounds. Metal concentrations were similar in male and female animals and were generally highest in mature animals. Liver concentrations of arsenic, chromium, iron, lead, manganese, mercury and nickel in a number of individual animals were elevated in comparison to concentrations previously reported in Australian dugong. Dieldrin, DDT (and its breakdown products) and/or heptachlor epoxide were detected in 59% of dugong blubber samples. In general, concentrations of organochlorines were similar to those reported in dugong 20 years earlier, and were low in comparison to concentrations recorded from marine mammal tissue collected elsewhere in the world. With the exception of lead, the extent of carcass decomposition, the presence of disease or evidence of animal starvation prior to death did not significantly affect dugong tissue concentrations of metals or organochlorines. The results of the study suggest that bioaccumulation of metals and organochlorine compounds (other than dioxins) does not represent a significant risk to Great Barrier Reef dugong populations, particularly in the context of other pressures associated with coastal development and other anthropogenic activities. PMID- 15757736 TI - Effects of the herbicide diuron on the early life history stages of coral. AB - The effects of the herbicide diuron on the early life history stages of broadcast spawning and brooding corals were examined in laboratory experiments. Fertilisation of Acropora millepora and Montipora aequituberculata oocytes were not inhibited at diuron concentrations of up to 1000 microg l(-1). Metamorphosis of symbiont-free A. millepora larvae was only significantly inhibited at 300 microg l(-1) diuron. Pocillopora damicornis larvae, which contain symbiotic dinoflagellates, were able to undergo metamorphosis after 24 h exposure to diuron at 1000 microg l(-1). Two-week old P. damicornis recruits on the other hand were as susceptible to diuron as adult colonies, with expulsion of symbiotic dinoflagellates (bleaching) evident at 10 microg l(-1) diuron after 96 h exposure. Reversible metamorphosis was observed at high diuron concentrations, with fully bleached polyps escaping from their skeletons. Pulse amplitude modulation (PAM) chlorophyll fluorescence techniques demonstrated a reduction in photosynthetic efficiency (Delta F/F(m)') in illuminated P. damicornis recruits after a 2 h exposure to 1 microg l(-1) diuron. The dark-adapted quantum yields (F(v)/F(m)) also declined, indicating chronic photoinhibition and damage to photosystem II. PMID- 15757737 TI - Changes in algal, coral and fish assemblages along water quality gradients on the inshore Great Barrier Reef. AB - Macroalgae, hard corals, octocorals, and fish were surveyed on 10 to 13 inshore coral reefs of the Great Barrier Reef, along a water quality gradient in two regions with contrasting agricultural land use. A water quality index was calculated for each reef based on available data of particulate and dissolved nutrients, chlorophyll and suspended solids. Strong gradients in ecological attributes occurred along the water quality gradient. Macroalgae of the divisions Rhodophyta and Chlorophyta increased with increasing nutrients, while Phaeophyta remained similar. Octocoral richness and abundances of many hard coral and octocoral taxa decreased, and none of the hundreds of species increased. At reefs in higher nutrient environments, hard coral and octocoral assemblages were composed of subsets of the many species found in lower nutrient environments, whereas fish and macroalgal assemblages consisted of contrasting suites of species. The study identifies species groups that are likely to increase or decrease in abundance with changing water quality. PMID- 15757738 TI - A demographic approach to monitoring the health of coral reefs. AB - Inshore coral reefs adjacent to the wet tropics in North Queensland, Australia, are regularly exposed to flood plumes from coastal river systems. Changes in the nature of these plumes have been linked to the declining health of coral reefs in the region. The effect of flood plumes on the health of inshore corals was investigated by quantifying aspects of the demography of populations of corymbose and digitate Acropora at three groups of Island reefs along a gradient of exposure and decreasing water quality (High Island >Frankland's >Fitzroy). The size-structures of colonies, the rates of sexual recruitment, and the growth and survival of juveniles, all varied among the Island reefs. Juvenile and adult sized colonies were far more abundant at the Fitzroy Island reefs, than at the High or Frankland Island reefs that were more exposed to flood plumes. Additionally, there were up to eight times as many sexual recruits at the Fitzroy Island reefs, compared with the High Island reefs. However, the rates of growth and survival of the juvenile sized corals at the Fitzroy Island reefs were lower than at the more exposed reefs. The comparatively low abundance of adult corals at the exposed reefs is most likely due to their histories of disturbance from crown-of-thorns and coral bleaching, but the lack of subsequent recovery due to their low levels of larval recruitment. If a stock-recruitment relationship is typical for these groups of reefs, then the low rates of recruitment may be linked to the low density of adult colonies. Alternately, direct or indirect effects of chronic exposure to poor water quality may have resulted in less suitable substrata for larval settlement. We discuss these results and provide examples of how information about population structure and dynamics can be used in simple matrix models to quantify the current and future health of populations of corals under various scenarios. PMID- 15757739 TI - Effects of algal turfs and sediment on coral settlement. AB - Successful settlement and recruitment of corals is critical to the resilience of coral reefs. Given that many degraded reefs are dominated by benthic algae, recovery of coral populations after bleaching and other disturbances requires successful settlement amidst benthic algae. Algal turfs often accumulate sediments, sediments are known to inhibit coral settlement, and reefs with high inputs of terrestrial sediments are often dominated by turfs. We investigated the impacts of two algal turf assemblages, and of sediment deposits, on settlement of the coral Acropora millepora (Ehrenberg). Adding sediment reduced coral settlement, but the effects of different algal turfs varied. In one case, algal turfs inhibited coral settlement, whereas the other turf only inhibited settlement when combined with sediments. These results provide the first direct, experimental evidence of effects of filamentous algal turfs on coral settlement, the variability in those effects, and the potential combined effects of algal turfs and trapped sediments. PMID- 15757740 TI - Synergistic effects of diuron and sedimentation on photosynthesis and survival of crustose coralline algae. AB - Effects of short-term exposure to sedimentation and diuron, separately and in combination, on the photophysiology and survival of crustose coralline algae (CCA) were examined in controlled time-course experiments, using pulse-amplitude modulation (PAM) chlorophyll fluorometry. These experiments indicated that the effects of sediments and diuron, when applied in isolation, were often reversible, with recovery time dependent upon sediment type and diuron concentration. Exposure to fine (<63 microm grain size), nutrient-rich estuarine sediments reduced effective quantum yields (Delta F/F(m')) of photosystem II in CCA species more than exposure to the same amount of fine (<63 microm grain size) calcareous sediments. Significant inhibition of photosynthesis (Delta F/F(m')) was also observed at diuron concentrations > or =2.9 microg L(-1). Fine estuarine sediments in combination with 0.79 microg L(-1) dissolved diuron, caused yields (Delta F/F(m')) to drop by 60% compared with controls after 24 h. The combined exposure to sediments and diuron also retarded recovery, thus Delta F/F(m') values were still only 60% of the controls after 9 days recovery in clean seawater. Mortality of CCA was observed in some fragments treated with combinations of sediment and diuron. Our results suggest that sediment deposition and exposure to diuron can negatively affect the photosynthetic activity of CCA, with sedimentation stress being significantly enhanced by the presence of trace concentrations of diuron. PMID- 15757741 TI - Blooms of the cyanobacterium Lyngbya majuscula in coastal Queensland, Australia: disparate sites, common factors. AB - During the last decade there has been a significant rise in observations of blooms of the toxic cyanobacterium Lyngbya majuscula along the east coast of Queensland, Australia. Whether the increase in cyanobacterial abundance is a biological indicator of widespread water quality degradation or also a function of other environmental change is unknown. A bioassay approach was used to assesses the potential for runoff from various land uses to stimulate productivity of L. majuscula. In Moreton Bay, L. majuscula productivity was significantly (p<0.05) stimulated by soil extracts, which were high in phosphorus, iron and organic carbon. Productivity of L. majuscula from the Great Barrier Reef was also significantly (p<0.05) elevated by iron and phosphorus rich extracts, in this case seabird guano adjacent to the bloom site. Hence, it is possible that other L. majuscula blooms are a result of similar stimulating factors (iron, phosphorus and organic carbon), delivered through different mechanisms. PMID- 15757742 TI - Influence of land runoff on rates and agents of bioerosion of coral substrates. AB - Annually large volumes of fresh water laden with sediment are washed down the Daintree River in North Queensland into the Great Barrier Reef lagoon. To investigate the effects of land runoff on bioerosion, samples of recently killed colonies of Porites were laid at 6 sites on a cross shelf transect from Snapper Island at the entrance to the river to Osprey Reef, approximately 328 km from the river mouth out in the Coral Sea. Rates and agents of bioerosion were determined over 4 years and inshore sites exhibited significantly lower rates of total bioerosion than the other sites. Offshore sites experienced high rates of bioerosion primarily due to grazing and internal bioerosion by macroborers such as sponges and bivalves was also important at some of these sites. Inshore sites were covered in heavy layers of silt which inhibited colonization and growth of microborers, primarily algae. This resulted in lower levels of grazing than at offshore sites. However the activity of macroborers (primarily sponges and bivalves) was often high at these sites. The macroboring communities differed between sites and over time and it is hypothesised that these site differences were due to different levels of terrestrial runoff. These results are compared with those from French Polynesia where contaminated terrestrial runoff greatly influenced rates and agents of bioerosion. However other factors such as overfishing may also play an important role in some locations. PMID- 15757743 TI - A risk assessment approach to contaminants in Port Curtis, Queensland, Australia. AB - Port Curtis is one of Australia's leading ports for which substantial industrial expansion is proposed over the next decade. However, there has been little attempt to date to assess the extent of contamination in waters, sediments and biota or to characterize the potential impacts of contaminants on aquatic biota. Contaminants of potential concern to biota and human health were investigated in the Port Curtis estuary using a screening-level risk assessment approach. Dissolved metal concentrations in waters were below [ANZECC/ARMCANZ, 2000. Australian and New Zealand Guidelines for Fresh and Marine Water Quality, Vol. 1. The Guidelines, Australian and New Zealand Environment and Conservation Council and Agriculture and Resource Management Council of Australia and New Zealand] trigger values, suggesting low risk of these contaminants. In sediments, arsenic, nickel and chromium concentrations exceeded interim sediment quality guidelines low (ISQG-low), but were also high in the reference zone suggesting a natural origin. Historical data on naphthalene in Harbour sediments showed that it was also of potential concern. Bioaccumulation of contaminants in a range of biota was also used as an indicator of contaminant exposure. Biota were generally enriched in metals and tributyltin, which was also elevated in water and sediments. Although not unique to Port Curtis, mercury in barramundi was identified as a potential risk to human health. PMID- 15757744 TI - Mapping water quality and substrate cover in optically complex coastal and reef waters: an integrated approach. AB - Sustainable management of coastal and coral reef environments requires regular collection of accurate information on recognized ecosystem health indicators. Satellite image data and derived maps of water column and substrate biophysical properties provide an opportunity to develop baseline mapping and monitoring programs for coastal and coral reef ecosystem health indicators. A significant challenge for satellite image data in coastal and coral reef water bodies is the mixture of both clear and turbid waters. A new approach is presented in this paper to enable production of water quality and substrate cover type maps, linked to a field based coastal ecosystem health indicator monitoring program, for use in turbid to clear coastal and coral reef waters. An optimized optical domain method was applied to map selected water quality (Secchi depth, Kd PAR, tripton, CDOM) and substrate cover type (seagrass, algae, sand) parameters. The approach is demonstrated using commercially available Landsat 7 Enhanced Thematic Mapper image data over a coastal embayment exhibiting the range of substrate cover types and water quality conditions commonly found in sub-tropical and tropical coastal environments. Spatially extensive and quantitative maps of selected water quality and substrate cover parameters were produced for the study site. These map products were refined by interactions with management agencies to suit the information requirements of their monitoring and management programs. PMID- 15757745 TI - Field assessment of innovative sensor for monitoring of sediment accumulation at inshore coral reefs. AB - Sediment accumulation rate is a frequently required parameter in environmental and management studies, in particular near coral reefs where sediment accumulation can potentially cause severe impact. However, opportunities to obtain accurate sediment accumulation measurements are often limited by a lack of adequate instrumentation, in particular for high temporal resolution monitoring. For instance the traditional use of sediment traps, as the most widespread technique, offers poor temporal resolution (commonly of weeks) besides having significant hydrodynamic shortcomings. Therefore, a new optical backscatter sediment accumulation sensor (SAS) was developed to continuously measure in situ short-term sediment accumulation in sensitive riverine and coastal environments, enabling high temporal and vertical resolution (order of 1 h and with a deposited thickness resolution in the order of 20 microm respectively). This allows investigations of various parameters that influence accumulation: tides, current, waves, rain, or anthropogenic activity such as sediment dumping. This paper briefly describes the SAS and presents three field applications on nearshore coral reefs at Ishigaki Island (Japan), Lihir Island (Papua New Guinea), and Magnetic Island (Australia). PMID- 15757746 TI - The development of a physiologically-based pharmacokinetic model using the distribution of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the tissues of the eastern oyster (Crassostrea virginica). AB - A physiologically-based pharmacokinetic model (PBPK) was developed to describe the kinetics of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the eastern oyster (Crassostrea virginica). The estimated t(1/2) of elimination for a bolus dose of TCDD in C. virginica is approximately 14-24 days based on both the experimental data and the PBPK model. The highest dioxin concentration reached during 28-days was in the digestive gland followed by the mantle, gonad, hemolymph, gill, adductor muscle, and the kidney/heart. A binding protein for 2,3,7,8-TCDD had been reported in the literature for both the digestive gland and gonad. Incorporating a binding component in the model resulted in a better fit for the data. The PBPK model predicted the distribution and the elimination concentrations for 2,3,7,8-TCDD within each of the tissue compartments. This model will serve as a useful tool for predicting the kinetics of other persistent organic pollutants as well as, allow for a more refined ecological risk assessment by estimating dioxin concentrations in sensitive tissues such as the gonad. PMID- 15757747 TI - Assessment of cytochrome P450 1A in harbour seals (Phoca vitulina) using a minimally-invasive biopsy approach. AB - Biomarkers of organochlorine exposure, such as the induction of cytochrome P450 1A (CYP1A), can be used to assess the impact of environmental contaminants on the health of free-ranging marine mammal populations. The objective of the present study was to measure CYP1A in skin and liver biopsies obtained from live harbour seals (Phoca vitulina). Twelve harbour seal pups, aged three to five weeks, were captured from the Fraser River estuary, British Columbia, Canada, and temporarily held in captivity. Skin ( approximately 60 mg) and liver ( approximately 40 mg) biopsies, obtained while seals were under general anaesthesia, yielded sufficient tissue for the measurement of CYP1A by immunoblot analysis and ethoxyresorufin O deethylase activity. A short-term exposure experiment, in which harbour seals (n=3) were treated orally with beta-naphthoflavone (BNF), resulted in increased hepatic and cutaneous CYP1A protein levels, consistent with observations in other mammals. This study is the first to measure CYP1A in skin and liver biopsies from live harbour seals and to report in vivo BNF-associated CYP1A induction in a marine mammal. The results demonstrate that microsamples collected using minimally-invasive techniques can provide toxicologically-relevant information form marine mammals. PMID- 15757748 TI - Three-dimensional hydrodynamic-eutrophication model (HEM-3D): application to Kwang-Yang Bay, Korea. AB - The purpose of this paper is twofold: to describe the water quality model of Three-Dimensional Hydrodynamic-Eutrophication Model (HEM-3D) and to present an application of HEM-3D to a coastal system in Korea. HEM-3D, listed as a tool for the development of Total Maximum Daily Load by US Environmental Protection Agency, is a general-purpose modeling package for simulation of the flow field, transport, and eutrophication processes throughout the water column and of diagenetic processes in the benthic sediment. This paper describes the water quality model of HEM-3D with emphasis on its unique features. Excessive loadings of organic wastes have significantly deteriorated water quality conditions of Korean coastal waters. This paper presents an application of HEM-3D to Kwang-Yang Bay, a coastal system in Korea, which is one of the first water quality modeling efforts for Korean coastal waters accompanied by a relatively comprehensive field program. The current status of data availability for water quality modeling in Korea is discussed. PMID- 15757749 TI - Extending the generality of ecological models to artificial floating habitats. AB - Marine assemblages on natural hard substrata are generally different from those on artificial habitats. There is, however, the potential for certain ecological processes to operate on both types of structures. On the sides of floating pontoons in Sydney Harbour, there were strong patterns of vertical distribution of sessile epibiotic organisms and molluscan grazers across relatively small spatial scales (in three defined zones, namely splash, shallow and deep). Patterns of vertical distribution of the tubeworms Hydroides spp. were reversed depending on the cover of mussels. A manipulative experiment was done to test if patterns of vertical distribution of Hydroides spp. were due to (1) the functioning of mussels or (2) the structure provided by mussels. Neither the functioning nor structure of mussels accounted for the patterns of distribution of Hydroides spp. Mussels increased recruitment of Hydroides spp., in the shallow and deep zones, and this was not due to increased surface area of the mussel shells. Manipulation of numbers of grazers and covers of sessile epibiota showed that the observed negative relationship between grazers and epibiota was due to grazers reducing recruitment of epibiota and epibiota decreasing survival of grazers. Most importantly, processes that accounted for patterns of distribution of mobile and sessile organisms on artificial floating structures were similar to those repeatedly shown to create such patterns on natural rocky shores. PMID- 15757750 TI - Environmental consequence analyses of fish farm emissions related to different scales and exemplified by data from the Baltic--a review. AB - The aim of this work is to review studies to evaluate how emissions from fish cage farms cause eutrophication effects in marine environments. The focus is on four different scales: (i) the conditions at the site of the farm, (ii) the local scale related to the coastal area where the farm is situated, (iii) the regional scale encompassing many coastal areas and (iv) the international scale including several regional coastal areas. The aim is to evaluate the role of nutrient emissions from fish farms in a general way, but all selected examples come from the Baltic Sea. An important part of this evaluation concerns the method to define the boundaries of a given coastal area. If this is done arbitrarily, one would obtain arbitrary results in the environmental consequence analysis. In this work, the boundary lines between the coast and the sea are drawn using GIS methods (geographical information systems) according to the topographical bottleneck method, which opens a way to determine many fundamental characteristics in the context of mass balance calculations. In mass balance modelling, the fluxes from the fish farm should be compared to other fluxes to, within and from coastal areas. Results collected in this study show that: (1) at the smallest scale (<1 ha), the "footprint" expressing the impact areas of fish cage farm often corresponds to the size of a "football field" (50-100 m) if the annual fish production is about 50 ton, (2) at the local scale (1 ha to 100 km2), there exists a simple load diagram (effect-load-sensitivity) to relate the environmental response and effects from a specific load from a fish cage farm. This makes it possible to obtain a first estimate of the maximum allowable fish production in a specific coastal area, (3) at the regional scale (100-10,000 km2), it is possible to create negative nutrient fluxes, i.e., use fish farming as a method to reduce the nutrient loading to the sea. The breaking point is to use more than about 1.1 g wet weight regionally caught wild fish per gram feed for the cultivated fish, and (4) at the international scale (>10,000 km2) related to the Baltic Proper, the contribution from fish farms to the overall nutrient fluxes are very small. We have also given two case-studies at the local scale where the impact of the fish farm emissions are greatest and the idea is to identify coastal areas unsuitable and suitable for fish cage farms and the reasons why. It should also be stressed that the results presented here are exemplified using emissions from fish farms, but that the underlying principles to evaluate the ecosystem effects of nutrient discharges from point source emissions are valid in a wider and more general perspective. PMID- 15757751 TI - Meiofauna as descriptor of tourism-induced changes at sandy beaches. AB - Tourism has long been considered as a 'clean industry' with almost no negative effects on the environment. This study demonstrated, in two different coastal systems (Mediterranean and Baltic), that tourism related activities are particularly affecting the sandy beach meio- and nematofauna in the upper beach zone, the specific ecotone in which many meiofauna species from both the marine and the terrestrial environment congregate. Tourist upper beaches are characterized by a lower % total organic matter (%TOM), lower densities, lower diversities (absence of Insecta, Harpacticoida, Oligochaeta, terrestrial nematodes and marine Ironidae nematodes) and higher community stress compared to nearby non-tourist locations. The %TOM was found to be the single most important factor for the observed differences in meiofauna assemblage structure at tourist versus non-tourist beaches in both the Mediterranean and the Baltic region. The free-living nematode assemblages from tourist upper zones depart significantly from expectations based on random selections from the regional nematode species pool. Furthermore upper zone assemblages are characterised by a low species diversity consisting of taxonomically closely related nematode species with r strategist features. Generally, faunal differences between tourist and non tourist beaches are decreasing towards the lower beach zones. PMID- 15757753 TI - Gene medicines: the end of the beginning? AB - First-generation gene medicines and genetic vaccines represent a promising new class of therapeutics that have the potential to prevent, correct, or modulate genetic or acquired diseases. The rational design of synthetic gene delivery and expression systems continues to be essential to enable the precise temporal and spatial control of transgene expression in vivo. With the tantalizing efficacy results and outstanding safety profile observed with nonviral, plasmid-based product candidates in early clinical trials, a multidisciplinary approach remains critical to further improve the effectiveness, reduce the manufacturing costs, and maintain the safety of gene therapeutics and vaccines for their successful development. This commentary provides an historical perspective on somatic gene therapy and briefly addresses the rate-limiting steps in effective gene transfer and expression. The importance of understanding plasmid pharmacokinetics after administration by conventional routes in animal models and in humans is emphasized. Pharmaceutical scientists have a pivotal role to play in deciphering the key biological parameters to effective gene transfer and designing gene delivery systems that will enable plasmid-based products to become an integral part of the future medical armamentarium. PMID- 15757754 TI - Theoretical considerations involving the pharmacokinetics of plasmid DNA. AB - Success of in vivo gene therapy relies on the development of gene delivery technologies, by which a well-controlled transgene expression is achieved as far as the spatial and temporal profile of the expression is concerned. Because transgene expression only occurs in cells that are transduced with the gene administered, the tissue distribution of genes is an important factor determining the efficacy of in vivo gene transfer. Plasmid DNA is the simplest vector and its administration in naked or complexed form results in significant transgene expression in various organs. The route of administration, the use of cationic vectors and the administration technique greatly affects the tissue distribution of plasmid DNA and the subsequent transgene expression. Therefore, a clear understanding of the tissue distribution of naked and complexed plasmid DNA is a prerequisite for strategies for developing effective in vivo gene transfer methods. Pharmacokinetics translates the tissue distribution properties of plasmid DNA into quantitative parameters, which can be compared with parameters obtained under different conditions, or with physiological parameters such as blood flow rate. Here we discuss the pharmacokinetic evaluation of the tissue distribution characteristics of plasmid DNA, in the free and complexed forms. PMID- 15757755 TI - Implications of pharmacokinetic behavior of lipoplex for its inflammatory toxicity. AB - Inflammatory toxicity represents a typical toxicity associated with systemic administration of cationic liposome/DNA complex (lipoplex). Collected information indicates that the lipoplex gene delivery system mediates an uptake of plasmid DNA by the liver, mainly by Kupffer cells, in which a large amount of cytokine is produced. Therefore, many efforts have been made to overcome this problem. Previous reports by our laboratory demonstrated that sequential injection of cationic liposome and DNA could dramatically decrease the toxicity. In comparison with lipoplex injection, this method significantly suppresses the uptake of DNA by the liver. Opsonization effect in the stimulation of Kupffer cell uptake is proposed as an explanation for the differences in the pharmacokinetic properties of plasmid DNA after lipoplex injection and sequential injection. In this review, we cover the current understanding of the mechanisms underlying inflammatory toxicity and the several attempts to overcome this toxicity. The mechanism related to the pharmacokinetic properties of the lipoplex is focused on here for discussion. PMID- 15757756 TI - Water insoluble and soluble lipids for gene delivery. AB - Among various synthetic gene carriers currently in use, liposomes composed of cationic lipids and co-lipids remain the most efficient transfection reagents. Physicochemical properties of lipid/plasmid complexes, such as cationic lipid structure, cationic lipid to co-lipid ratio, charge ratio, particle size and zeta potential have significant influence on gene expression and biodistribution. However, most cationic lipids are toxic and cationic liposomes/plasmid complexes do not disperse well inside the target tissues because of their large particle size. To overcome the problems associated with cationic lipids, we designed water soluble lipopolymers for gene delivery to various cells and tissues. This review provides a critical discussion on how the components of water insoluble and soluble lipids affect their transfection efficiency and biodistribution of lipid/plasmid complexes. PMID- 15757757 TI - The hydrodynamics-based procedure for controlling the pharmacokinetics of gene medicines at whole body, organ and cellular levels. AB - Hydrodynamics-based gene delivery, involving a large-volume and high-speed intravenous injection of naked plasmid DNA (pDNA), gives a significantly high level of transgene expression in vivo. This has attracted a lot of attention and has been used very frequently as an efficient, simple and convenient transfection method for laboratory animals. Until recently, however, little information has been published on the pharmacokinetics of the injected DNA molecules and of the detailed mechanisms underlying the efficient gene transfer. We and other groups have very recently demonstrated that the mechanism for the hydrodynamics-based gene transfer would involve, in part, the direct cytosolic delivery of pDNA through the cell membrane due to transiently enhanced permeability. Along with the findings in our series of studies, this article reviews the cumulative reports and other intriguing information on the controlled pharmacokinetics of naked pDNA in the hydrodynamics-based gene delivery. In addition, we describe various applications reported so far, as well as the current attempts and proposals to develop novel gene medicines for future gene therapy using the concept of the hydrodynamics-based procedure. Furthermore, the issues associated with the clinical feasibility of its seemingly invasive nature, which is probably the most common concern about this hydrodynamics-based procedure, are discussed along with its future prospects and challenges. PMID- 15757758 TI - Physical methods for gene transfer: improving the kinetics of gene delivery into cells. AB - One factor critical to successful gene therapy is the development of efficient delivery systems. Although advances in gene transfer technology, including viral and non-viral vectors, have been made, an ideal vector system has not yet been constructed. This review describes the basic principles behind various physical methods for gene transfer and assesses the advantages and performance of such approaches, compared to other transfection systems. In particular, the kinetics and efficiency of gene delivery, the toxicity, in vivo feasibility, and targeting ability of different physical methodologies are discussed and evaluated. PMID- 15757759 TI - Intracellular routing of plasmid DNA during non-viral gene transfer. AB - Gene transfer using non-viral vectors is a promising approach for the safe delivery of therapeutic DNA in genetic and acquired human diseases. Whereas the lack of specific immune response favors the use of plasmid-cationic polymer complexes, the limited efficacy and short duration of transgene expression impose major hurdles in the application of non-viral gene delivery techniques. Here, we review the major cellular, metabolic and physico-chemical impediments that non viral vectors encounter before plasmid DNA enters the nucleus. Following endocytosis of DNA-polycation complexes, a large fraction of the DNA is targeted to the lysosomes. Since the cytosolic release of heterologous DNA is a prerequisite for nuclear translocation, entrapment and degradation of plasmid DNA in endo-lysosomes constitute one of the major impediments to efficient gene transfer. Plasmid DNA that escapes the endo-lysosomal compartment encounters the diffusional and metabolic barriers of the cytoplasm, reducing greatly the number of intact plasmids that reach the nucleosol. Nuclear translocation of DNA requires either the disassembly of the nuclear envelope or active nuclear transport via the nuclear pore complex. A better understanding of the cellular and molecular basis of non-viral vector trafficking from the extracellular compartment into the nucleus may provide strategies to overcome those obstacles that limit the efficiency of gene delivery. PMID- 15757760 TI - Controlling the kinetics of transgene expression by plasmid design. AB - While a vast array of liposomes, peptides, and molecular conjugates have been evaluated for nonviral gene transfer, the entity containing the actual gene itself is almost always a plasmid. The layout of most plasmid DNA (pDNA) vectors is usually quite simple, consisting of a promoter, transgene, polyadenylation signal, and a backbone that permits propagation of the plasmid in bacteria. Additional sequence elements and modifications can be incorporated to influence the stability of gene expression and retention of the pDNA molecule in a given tissue. This review describes the different choices that can be made when designing a pDNA vector for transient, sustained, or regulated expression. The choice of promoter is a major determinant governing the kinetics of expression, but other factors, such as CpG content and the topological form of the pDNA are also influential. Vectors can also be designed to respond to the local environment of a given cell or tissue, or engineered to respond to a small molecule drug. PMID- 15757761 TI - Approaches to improving the kinetics of adenovirus-delivered genes and gene products. AB - Adenovirus (Ad) vectors have been expected to play a great role in gene therapy because of their extremely high transduction efficiency and wide tropism. However, due to the intrinsic deficiency of their immunogenic toxicities, Ad vectors are rapidly cleared from the host, transgene expression is transient, and readministration of the same serotype Ad vectors is problematic. As a result, Ad vectors are continually undergoing refinement to realize their potential for gene therapy application. Even after 1999, when a patient fatally succumbed to the toxicity associated with Ad vector administration at a University of Pennsylvania (U.S.) experimental clinic, enthusiasm of gene therapists for Ad vectors has not waned. With great efforts from various research groups, significant advances have been achieved through comprehensive approaches to improving the kinetics of Ad vector-delivered genes and gene products. PMID- 15757762 TI - Dorsal fracture-dislocation of the proximal interphalangeal joint: a comparative study of percutaneous Kirschner wire fixation versus open reduction and internal fixation. AB - Nineteen patients with a dorsal fracture-dislocation of the proximal interphalangeal joint of a finger were treated with either closed reduction and transarticular Kirschner wire fixation (eight cases) or open reduction and internal fixation, using either one or two lag screws (six cases) or a cerclage wire (five cases). At a mean follow-up of 7 (range 6-9) years, most patients reported satisfactory finger function, even though some of the injuries healed with proximal interphalangeal joint incongruency (seven cases) or subluxation (four cases). Those treated by open reduction complained of more "loss of feeling" in the affected finger and those specifically treated by cerclage wire fixation reported more cold intolerance and had a significantly larger fixed flexion deformity (median, 30 degrees : range 18-38 degrees ) and a smaller arc of motion (median, 48 degrees : range 45-60 degrees ) at the proximal interphalangeal joint, despite having the best radiological outcomes. Closed reduction and transarticular Kirschner wire fixation produced satisfactory results, with none of the eight patients experiencing significant persistent symptoms despite a reduced arc of proximal interphalangeal joint flexion (median=75 degrees ; range 60-108 degrees ). The results of this relatively simple treatment appear at least as satisfactory as those obtained by the two techniques of open reduction and internal fixation, both of which were technically demanding. PMID- 15757763 TI - Results of ulnar shortening osteotomy for ulnocarpal impaction syndrome in wrists with neutral or negative ulnar variance. AB - Ulnocarpal impaction syndrome was diagnosed in six wrists of five patients with neutral or negative ulnar variance. All underwent ulnar shortening with satisfactory results. The average grip strength increased from 53% to 78% and the range of flexion-extension increased from 82% to 93%, the mean Cooney's score improved from 25 to 83. These cases show that ulnocarpal impaction syndrome can occur in wrists with zero or negative ulnar variance, and that ulnar shortening is an effective treatment for such wrists. PMID- 15757764 TI - Bilateral Kienbock's disease. AB - We conducted a retrospective review of 11 patients with bilateral Kienbock's disease from our series of 251 patients with Kienbock's disease. There were no significant differences in radiographic parameters, including ulnar variance and carpal bone angle, between those with unilateral and those with bilateral Kienbock's disease. None of the patients with bilateral disease had been treated with corticosteroids or had a systemic disease that predisposed to osteonecrosis. Thus, this study failed to demonstrate any risk factor for bilateral, as opposed to unilateral Kienbock's disease. PMID- 15757765 TI - Scapholunate ligament injury occurring with scaphoid fracture--a rare coincidence? AB - In a retrospective study the results of a series 34 wrist arthroscopies in 189 acute scaphoid fractures were analysed. Scapholunate ligament tears were found in 13 cases. In 10 cases there was complete disruption of scapholunate interosseous ligament causing dynamic instability. Partial ligament tears without instability were seen in three cases. Despite the limitations of this series we conclude that occurrence of scapholunate ligament injury with a scaphoid fracture may be more common than generally thought. PMID- 15757766 TI - Functional anatomy of the lateral collateral ligament complex of the elbow: morphology and strain. AB - The anatomy of the lateral ulnar collateral ligament (LUCL) of the elbow was investigated in 26 fresh frozen cadavers. Two types of insertion of the LUCL were originally described but we found another type which is characterized by a broad single expansion along with a thin membranous fibre. Strain on the LUCL was measured in situ during extension and flexion with the forearm in supination, pronation and neutral. Strain in the proximal fibres started to occur at around 32 degrees flexion and peaked at between 50 degrees and 60 degrees flexion. Strains measured in the distal fibres were smaller in magnitude. Forearm rotation had little effect on strain during extension to flexion. Based on these results, we conclude that the LUCL functions in unison with the annular ligament. PMID- 15757767 TI - Dupuytren's disease secondary to acute injury, infection or operation distal to the elbow in the ipsilateral upper limb--a historical review. AB - The aggregated total of 385 cases of Dupuytren's disease arising after acute or specific injury, operation or infection of the forearm, wrist or hand between 1614 and 2003 are documented, including a personal series of 52 cases. The history of this relationship is recorded and the medico legal implications of the association are discussed. PMID- 15757768 TI - The effect of forearm bracing on radioulnar impingement. AB - The purpose of this clinical study was to compare patients' symptoms and radioulnar convergence following distal radioulnar reconstruction with and without a functional brace on. Standardized radiographs were taken of the involved wrist at rest, while holding a weight, and during maximum power grip. All radiographs were then repeated while wearing a prefabricated functional brace (Maramed, Miami, FL). The functional brace did not decrease radioulnar convergence and, at rest, the brace actually increased radioulnar convergence. Despite this, all patients who experienced pain without the brace were pain free with the brace on. We conclude that forearm bracing may be a useful nonoperative treatment for residual pain following DRUJ salvage surgery, though its mechanism of action is unclear. PMID- 15757770 TI - Differential splintage for flexor tendon rehabilitation: an experimental study of its effect on finger flexion strength. AB - We conducted laboratory tests to investigate the possibility of partly de powering flexor digitorum profundus with a view of reducing flexion force during active flexor tendon rehabilitation. We constructed a splint and applied tapes to the proximal segments of fingers to test the hypothesis that holding three fingers more extended than the other finger would reduce the flexion strength of the more flexed finger. The splint allowed the metacarpophalangeal joint of the more flexed finger to be held in three positions of increasing flexion (15 degrees , 30 degrees , and 45 degrees ) compared to the remaining three fingers. We have called this 'differential splintage'. Healthy volunteers were tested for maximum active flexion strength at the different flexion angles. 'Differential splintage' of up to 45 degrees resulted in mean decreased flexion strength of 28% in the index finger and 35% to 38% in the middle, ring and little fingers. The results suggest that "differential splintage" of a finger after flexor tendon repair may be useful in reducing tension across the repair during a program of active tendon rehabilitation and we feel that it has potential to reduce the incidence of repair rupture before healing is complete. PMID- 15757769 TI - The role of antibiotic prophylaxis in clean incised hand injuries: a prospective randomized placebo controlled double blind trial. AB - A prospective, randomized, double blind, placebo controlled trial was designed to investigate the effect of prophylactic flucloxacillin on the infection rate in clean incised hand injuries, which included trauma to skin, tendon and nerve in adults. Using strict exclusion criteria, a total of 170 patients were recruited into one of three trial groups; Group A - intravenous flucloxacillin on induction followed by an oral placebo; Group B - intravenous flucloxacillin on induction followed by an oral flucloxacillin course or Group C - oral placebo. Thirteen of the patients were subsequently withdrawn, leaving 92% available to complete the trial. Infection was diagnosed using clinical criteria. The infection rates in the three groups were Group A - 13%, Group B - 4% and Group C - 15%. Strictly, the results demonstrate no statistically significant difference in the infection rates between the groups. PMID- 15757771 TI - Extensor tendon rehabilitation a prospective trial comparing three rehabilitation regimes. AB - Forty-two patients with 46 complete extensor tendon injuries were prospectively allocated to one of three rehabilitation regimes: static splintage; interphalangeal joint mobilization with metacarpophalangeal joint immobilization or; the "Norwich" regime. All 42 patients were operated on by one surgeon and assessed by one hand therapist. At 4 weeks the total active motion in the static splintage group was significantly reduced but by 12 weeks there was no difference between the regimes. There was no difference in total active motion between the repaired and uninjured hand at 12 weeks, with all patients achieving good or excellent results. However, grip strength at 12 weeks was significantly reduced compared to the uninjured hand after static splintage. There was no difference in hand therapy input between the regimes. PMID- 15757772 TI - Delayed single Kirschner wire compression technique for mallet fracture. AB - Twenty-four mallet fractures which involved at least one third% of the articular surface of the distal interphalangeal joint were treated by open reduction and internal fixation using a single double-ended Kirschner wire at a mean of 9 days after injury (range 4-15). At a mean follow-up of 43 (range 12-18) months the active range of motion, pain and the Warren and Norris criteria were evaluated. The mean active range of motion was from - 2 degrees extension (range 0-10 degrees ) to 72 degrees flexion (range 50-90 degrees ). Nineteen patients were pain free and five suffered from mild pain during strenuous work. The Warren and Norris results were successful in 22 and improved in two cases. Radiographs showed, that all the fractures united in a near-anatomic position but with joint narrowing in six digits. PMID- 15757773 TI - Anatomic reconstruction of the distal radioulnar ligaments: long-term results. AB - Chronic post-traumatic dynamic distal radioulnar joint instability following rupture of the distal radioulnar ligaments may cause disabling and progressive ulnar-sided wrist symptoms. We use a free tendon graft to anatomically reconstruct both dorsal and palmar distal radioulnar ligaments simultaneously. Nine patients who underwent this procedure between 1990 and 1997 were assessed using a modified Mayo Wrist Score. Following surgery, average wrist scores improved from 66 (95% CI, +/-3.0) to 92 (95% CI, 92+/-6.9) and were maintained in the long term (average score, 87 at 9 years). Joint stability was restored and maintained in seven of the nine patients, but two developed recurrent instability. Pain, grip strength and function were better following surgery in the short and long term. Range of supination and pronation decreased slightly from an average of 169 degrees pre-operatively to 155 degrees (90% of normal) in the long term. Long-term radiographs show no distal radioulnar or ulnocarpal joint osteoarthritis. PMID- 15757774 TI - Full thickness skin graft for nail unit reconstruction. AB - A retrospective study of 13 patients assessed the use of a full thickness skin graft for nail unit reconstruction after total nail unit removal for nail bed malignancies. No failures of the graft were observed and no patient had recurrence of the malignant tumour at 4 year follow-up. Full thickness skin grafting is a simple procedure which provides a good cosmetic outcome and does not produce significant donor site morbidity. PMID- 15757775 TI - Carpal tunnel syndrome in association with hand-arm vibration syndrome: a review of claimants seeking compensation in the Mining Industry. AB - Twenty six thousand eight hundred and forty-two miners seeking compensation were clinically assessed for vascular and neurosensory impairment arising from exposure to occupational hand-arm vibration (Hand-Arm Vibration Syndrome). They were also assessed clinically for Carpal Tunnel Syndrome which, if present, would result in additional compensation. Fifteen per cent were assessed as having both HAVS and CTS. Thirty-eight per cent of claimants had nocturnal wakening, 1.3% wasting of abductor pollicis brevis, 15% had a positive Tinel's test and 20% had a positive Phalen's test. The 15% prevalence reported is lower than the rates cited previously in several small population studies of workers exposed to vibration. This paper reports the results of the assessment process and discusses the difficulty of discriminating Carpal Tunnel Syndrome from diffuse neurosensory impairment arising from HAVS. PMID- 15757776 TI - Physical examination of the carpal bones by orthopaedic and accident and emergency surgeons. AB - Our aim was to test knowledge of carpal bone surface anatomy among orthopaedic and accident and emergency surgeons. A survey of 58 surgeons was conducted. Each was asked to palpate seven points on five commonly injured carpal bones. Eight surgeons declined to participate. Five of the remaining 50 correctly palpated all surface markings. Six failed to accurately palpate even a single point. The scaphoid waist and trapezial ridge were accurately palpated by 47 surgeons. The median score for all participants was 3 correctly identified points. The majority of orthopaedic and accident and emergency surgeons do not perform an accurate physical examination of the carpal bones. The ancient art of history taking, precise physical examination and elucidation of a confident diagnosis is at risk of being replaced with tentative diagnoses and imaging techniques that are expensive and time consuming. We believe that basic anatomical teaching is inadequate. PMID- 15757777 TI - Manual Ability Measure (MAM-16): a preliminary report on a new patient-centred and task-oriented outcome measure of hand function. AB - The purpose of this study was to develop an easy-to-use and psychometrically sound outcome instrument that is task-oriented and patient-centred. One hundred fifteen patients with a variety of hand impairments completed a rating scale of perceived manual ability (i.e., the Manual Ability Measure). The first 70 patients also completed two other questionnaires about physical health and psychological well-being. Rasch Analyses were conducted to transform the ordinal ratings into linear measures; Rasch statistics were used to evaluate its measurement properties at both scale and item levels. Eighty-three original items were reduced to 16 common tasks; Rasch reliabilities were good; the easy-to difficult item hierarchy makes sense clinically. Moderate correlations were found between manual ability, physical function and general sense of well-being. The results of this preliminary study suggest that the MAM is a promising outcome measure that has adequate psychometric properties and can be used to complement other objective clinical measurements. PMID- 15757778 TI - Total wrist fusion: a study of 115 patients. AB - This retrospective study evaluated the outcome of total wrist fusion, predominantly using the Mannerfelt technique, in patients with rheumatoid arthritis. One hundred and fifteen patients were operated on for painful wrist destruction. The mean pain scores were 3.6 (1-4) pre-operatively, 1.9 (1-4) after 6 weeks and 1.3 (1-4) at 1 year. Although the radiological fusion rate was good, only 40% of the patients were very satisfied with the result. Ability to perform activities of daily living was only very much improved in 30% of the patients. Grip strength significantly improved after surgery. PMID- 15757779 TI - Minimally invasive plate osteosynthesis for comminuted fractures of the metaphysis of the radius. AB - Five comminuted and displaced fractures of the distal metaphysis of the radius were treated by a technique of minimally invasive plate osteosynthesis (MIPO) with the aim of minimizing soft tissue damage and devascularization of the fracture fragments. This technique used the small AO T-shaped locking compression plate (AO LCP T-plate) and left the pronator quadratus intact. Radial inclination, palmar tilt, and ulnar variance were restored without loss of reduction in all five cases and the fractures healed at an average of 10 weeks, with good to excellent clinical outcomes. PMID- 15757780 TI - Long-term outcome of radial shortening with or without ulnar shortening for treatment of Kienbock's disease: a 13-25 year follow-up. AB - Fourteen of 35 patients who underwent radial shortening with or without ulnar shortening for the treatment of Kienbock's disease were followed up for a median of 19 (range, 13-25) years. Radial shortening was performed for patients with ulnar negative or neutral variance, and combined shortening of radius and ulna for those with ulnar positive variance. Overall the clinical situation was significantly improved at the final follow-up. There was no significant advanced collapse of the wrists and Lichtman's stage of disease increased in only three cases. Although osteoarthritic changes in the distal radio-ulnar joint progressed in five patients, this may have little influence on clinical outcome. Radial shortening osteotomy is a reliable method for treatment of Kienbock's disease. PMID- 15757781 TI - Histoplasmosis infection presenting as an isolated subcutaneous periarticular upper limb swelling in the immunosuppressed patient. AB - Atypical mycobacterial and fungal infections may occur in immunosuppressed patients. The impaired host response can make the clinical presentation atypical. Blood and tissue cultures may be negative in the acute phase of the illness, which can lead to a delay in diagnosis. In those patients with AIDS or other underlying immunosuppressive conditions, histoplasmosis cannot always be eradicated, but treatment that achieves chronic suppression may be adequate to maintain functional capacity. This report describes two immunosuppressed patients who presented with isolated subcutaneous histoplasmosis infection around the wrist. PMID- 15757782 TI - Ischaemic muscle paralyses and contractures. PMID- 15757784 TI - Pain scores for patients undergoing open carpal tunnel decompression under local anaesthetic with plain or buffered lidocaine. PMID- 15757786 TI - Overview of molecular, cellular, and genetic neurotoxicology. AB - It has become increasingly evident that the field of neurotoxicology is not only rapidly growing but also rapidly evolving, especially over the last 20 years. As the number of drugs and environmental and bacterial/viral agents with potential neurotoxic properties has grown, the need for additional testing has increased. Only recently has the technology advanced to a level that neurotoxicologic studies can be performed without operating in a "black box." Examination of the effects of agents that are suspected of being toxic can occur on the molecular (protein-protein), cellular (biomarkers, neuronal function), and genetic (polymorphisms) level. Together, these areas help to elucidate the potential toxic profiles of unknown (and in some cases, known) agents. The area of proteomics is one of the fastest growing areas in science and particularly applicable to neurotoxicology. Lubec et al, provide a review of the potential and limitations of proteomics. Proteomics focuses on a more comprehensive view of cellular proteins and provides considerably more information about the effects of toxins on the CNS. Proteomics can be classified into three different focuses: post-translational modification, protein-expression profiling, and protein network mapping. Together, these methods represent a more complete and powerful image of protein modifications following potential toxin exposure. Cellular neurotoxicology involves many cellular processes including alterations in cellular energy homeostasis, ion homeostasis, intracellular signaling function, and neurotransmitter release, uptake, and storage. The greatest hurdle in cellular neurotoxicology has been the discovery of appropriate biomarkers that are reliable, reproducible, and easy to obtain. There are biomarkers of exposure effect, and susceptibility. Finding the appropriate biomarker for a particular toxin is a daunting task. The appropriate biomarker for a particular toxin is a daunting task. The advantage to biomarker/toxin combinations is they can be detected and measured shortly following exposure and before overt neuroanatomic damage or lesions. Intervention at this point, shortly following exposure, may prevent or at least attenuate further damage to the individual. The use of peripheral biomarkers to assess toxin damage in the CNS has numerous advantages: time-course analysis may be performed, ethical concerns with the use of human subjects can partially be avoided, procedures to acquire samples are less invasive, and in general, peripheral studies are easier to perform. Genetic neurotoxicology comprises two focuses--toxin-induced alterations in genetic expression and genetic alterations that affect toxin metabolism, distribution, and clearance. These differences can be beneficial or toxic. Polymorphisms have been shown to result in altered metabolism of certain toxins (paraoxonase and paraoxon). Conversely, it is possible that some polymorphisms may be beneficial and help prevent the formation of a toxic by-product of an exogenous agent (resistance to ozone-induced lung inflammation). It has also become clear that interactions of potential toxins are not straightforward as interactions with DNA, causing mutations. There are numerous agents that cause epigenetic responses (cellular alterations that are not mutagenic or cytotoxic). This finding suggests that many agents that may originally have been thought of as nontoxic should be re-examined for potential "indirect" toxicity. With the advancement of the human genome project and the development of a human genome map, the effects of potential toxins on single or multiple genes can be identified. Although collectively, the field of neurotoxicology has recently come a long way, it still has a long way to go reach its full potential. As technology and methodology advances continue and cooperation with other disciplines such as neuroscience, biochemistry, neurophysiology, and molecular biology is improved, the mechanisms of toxin action will be further elucidated. With this increased understanding will come improved clinical interventions to prevent neuronal damage following exposure to a toxin. PMID- 15757787 TI - Neurodevelopmental toxicology. AB - The fields of neurotoxicology and developmental toxicology are exploding in research and interest. Much of the data currently known are from epidemiologic human studies or studies of animal models. Each of these modes is difficult to translate to individual clinical encounters. It is often difficult to state with certainty which of the numerous chemical or physical agents in our environment are neurotoxic. Basic scientists will help with advances in molecular biology and toxicology. Improved clinical understanding of these issues may help patients to understand the medical issues; allay feelings of anxiety, guilt, or fear; and avoid unnecessary testing. For exposures that manifest as threshold phenomena, such as lead, the risk to society is even greater than to an individual. Individual risk may be less of a concern than the population's risk because small elevations in the average BLL can cause profound shifts in the normative curve of intelligence, increasing the burden on our institutions and bankrupting the brain trust. Good scholarship and interpersonal judgement are vital when counseling patients on the potential consequences of chemical exposures and are no less important when making policy. The challenge for the clinician reading the research is to remain aware of the limitations and biases of our science. PMID- 15757788 TI - Historical neurotoxins: what we have learned from toxins of the past about diseases of the present. AB - Throughout history, humans have fallen victim to a variety of neurotoxins, with exposures coming in the form of tainted products, industrial pollution, drugs of abuse, and even the bread and water that sustain them. Despite this long and tumultuous history, neurotoxic outbreaks still occur with regular frequency. Although many difficulties currently exist in linking many of today's unexplained neurologic disorders to toxins, the past suggests a prominent role for neurotoxins in diseases (such as amyotrophic lateral sclerosis and PD), unexplained peripheral neuropathies, neurodevelopmental disorders, and many psychiatric disturbances. PMID- 15757789 TI - Toxic encephalopathies. AB - Effective brain function is dependent on precise and complex interactions among neurotransmitters, hormones, enzymes, and electrolytes. Many of the chemically complex substances with which we come into contact can disrupt this intricately balanced system. Toxic substances, whether ingested, inhaled, or absorbed through the skin, may cause an encephalopathic state directly by affecting the brain itself or indirectly by compromising the brain's supportive systems. The article focuses on neurotoxins (heavy metals, solvents and vapors, pesticides, and natural neurotoxins) that directly induce an encephalopathic state. PMID- 15757790 TI - Toxic neuropathies. AB - Many substances, organic and manufactured, may induce peripheral nerve damage when exposed to them. The expected clinical phenotype is of a distal, sensory or sensorimotor polyneuropathy, often painful, with axonal characteristics on electrodiagnostic and histopathologic analysis. Treatment is limited; often, the only effective management is supportive care and avoidance from or removal of the offending toxin. Fortunately, the majority of toxic neuropathies are self-limited and improves gradually after toxin elimination. PMID- 15757791 TI - Toxic myopathies. AB - Muscle tissue is highly sensitive to drugs and toxins because ot its high metabolic activity and potential sites for disruption of energy-producing pathways. Early recognition of toxic myopathies is important, as they potentially are reversible on removal of the offending toxin, with greater likelihood of complete resolution the sooner this is achieved. Clinical features range from mild muscle pain and cramps to severe weakness with rhabdomyolysis, renal failure, and even death. The pathogenic bases can be multifactorial. This article reviews drugs responsible for common types of toxic myopathy and their clinical and histopathologic features and illustrates possible underlying cellular mechanisms. PMID- 15757792 TI - Toxin-induced movement disorders. AB - Toxins can be cited as a cause of several movement disorders, but this association is rare and the resultant syndromes usually include additional signs that are not typical for the idiopathic movement disorders. Most instances of confirmed toxin-induced movement disorders show lesions on CT and MRI scans of cortical or subcortical structures. A common underlying element in these toxin induced syndromes is the development of lesions primarily in the pallidum and striatum. Because many toxins result in lesions affecting these structures, a selective vulnerability to hypoxic or metabolic insults has long been postulated. The susceptibility of these structures may relate to a number of factors, including the pattern of oxidative metabolism, heavy metal concentration, vascular perfusion, and neuronal innervation. Finally, in addition to causing disability, certain neurotoxins have led to a better understanding of human disease through the development of research models. As an example, the MPTP model has not only provided an animal model to study therapeutic strategies in PD but has also contributed important insights into the mechanism of neuronal degeneration. PMID- 15757793 TI - Amyotrophic lateral sclerosis: possible role of environmental influences. AB - This treatise briefly discusses the genetic features of ALS and reviews environmental exposures in sporadic ALS. At least 10 genetic foci are responsible for cases of familial motor neuron disease and more are yet to be discovered. Research into sporadic ALS suggests that abundant factors apparently participate in the disease process. A singular cause and unifying disease and nerve dysfunction in polyneuropathies, a multitude of genetic, toxic, autoimmune, infectious, and systematic processes seem to be at play. The ALS syndrome likely will not be dissimilar. PMID- 15757794 TI - Neurodegenerative memory disorders: a potential role of environmental toxins. AB - The hypothesis that neurotoxins may play a role in neurodegenerative disorders remains an elusive one, given that epidemiologic studies often provide conflicting results. Although these conflicting results may result from methodological differences within and between studies, the complexity of chemical disruption of the central nervous system cannot be ignored in attempts to evaluate this hypothesis in different neurodegenerative disorders. Spencer provides a detailed review of the complex processes involved in defining the neurotoxic potential of naturally occurring and synthetic agents. Even concepts such as exposure and dose, as often reported in studies attempting to evaluate the risk imparted by a potential compound, can be deceptive. For example, although dose reflects "that amount of chemical transferred to the exposed subject", factors such as time and concentration in the organism, the ability to access the central nervous system, and how a compound reaches the central nervous system (routes of administration) or secondarily affects other organ systems leading to central nervous system disruption are clearly important to the concept of neurotoxic risk in neurodegenerative disorders. These factors would appear to explain the observed disagreements between studies using animal or neuronal models of neurotoxicity and population-based studies in humans. The importance of these factors and how a potential neurotoxin is investigated are clearly seen in the data on AD and aluminum. In contrast, the impact of MTPT on the central nervous system is more direct and compelling. Added complexity in the study of neurotoxins in human neurodegeneration is derived from data showing that agents may have additive, potentiating, synergistic, or antagonistic effects. Therefore, data from studies evaluating EMF risks could be readily confounded by the presence or absence of heavy metals (eg, arc welding). Other factors that may conceal neurotoxic causes for a given disorder focus on additional features such as genetic predispositions, physiologic changes that occur in aging, and even nutritional status that can support or hinder the affect of a given agent on the central nervous system. Finally, many studies that investigate exposure risk do not readily incorporate the five criteria proposed by Schaumburg for establishing causation. For example, if we apply Schaumburg's first criterion, epidemiologic studies often determines the presence of an agent through history, yet they cannot readily confirm exposure based on environmental or clinical chemical analyses to fulfill this criterion for causation. Additional limitations in research design along with the populations and methods that are sued to study neurotoxins in human neurodegenerative disorders often fail to meet other criteria such as linking the severity and onset with duration and exposure level. Therefore, although studies of agents such as MTPT provide compelling models of neurotoxins and neurodegeneration in humans, disorders such as ALS, PD, and particularly AD will require additional effort if research is to determine the contribution (presence or absence) of neurotoxins to these neurologic disorders. PMID- 15757795 TI - Gulf war syndrome: a toxic exposure? A systematic review. AB - Using the strength-of-conclusion scheme enumerated in Box 2, based on two class II studies, there is probably a causal link between deployment to the Persian Gulf theater of operation and the development of the poorly defined multisymptom illness known as GWS (level B). Based on class IV studies, there is insufficient evidence to determine if exposure to toxins encountered during the Persian Gulf war caused GWS (level U). A major limitation of the literature regarding the GWS is the reliance on self-reporting to measure exposure to putative causal toxins. Although objective measures of toxin exposure in GWV generally is unavailable, modeling techniques to estimate exposure levels to low-level nerve agents and smoke from oil well fires have been developed. It would be useful to determine if exposure levels determined by these techniques are associated with GWS. The lack of a clear case definition GWS also hampers research. Some go even further, claiming that the absence of such a definition renders the condition illegitimate. Although an objective marker to GWS would be useful for studies, the absence of such a marker does not make the syndrome any less legitimate. in essence, GWS merely is a convenient descriptive term that describes a phenomenon: GWV reporting suffering from medically unexplained health-related symptoms. In this sense, it shares much with the other medically unexplained syndromes encountered in practice. The real debate surrounding medically unexplained conditions is not whether or not they exist, but defining their cause. In this regard, investigators fall into two camps. One camp insists that the conditions are caused by a yet-to-be-discovered medical problem, rejecting out of hand the possibility of a psychologic origin. The other camp insists the conditions are fundamentally psychogenic rejecting the possibility of an undiscovered medical condition. The evidence shows, however, that the conditions exists, the suffering is real, and the causes are unknown. PMID- 15757796 TI - The neurology of aviation, underwater, and space environments. AB - It can be seen that aspects of different environments can have adverse effects on normal, healthy nervous systems. Aerospace and underwater neurology consultants are often required to evaluate aviators, divers, or astronauts who have neurologic problem to determine whether they should be granted a waiver. In general, these considerations include course of the disorder (static, progressive, or paroxysmal), potential for sudden incapacitation that may compromise safety, predictability of course of the disorder, ability to monitor disease, and potential adverse effects of medications. A background in the toxic effects of an abnormal environment on the nervous system can be used to make better clinical judgements when considering effects of exposing someone with an abnormal nervous system to such environmental stressors. PMID- 15757797 TI - Wilderness and recreational neurology. AB - With increased access to once remote regions of the planet and renewed interest in exploring natural surroundings, previously geographically isolated and rare neurologic conditions can be encountered in any patient population. Rare envenomations and poisonings, once the purview of the tropical neurologist, now can be encountered by travelers to areas where creatures that have developed specialized defenses are endemic. Recognition, therefore, of the potentially neurotoxic fauna and flora in these areas holds value, even for the urban neurologist. PMID- 15757798 TI - Radiation as a nervous system toxin. AB - Neurotoxicity from radiation can range widely and produce effects that may include (1) small absolute increases in cancer risks, (2) subtle effects on higher level functioning in some individuals, (3) severe cognitive impairment in some individuals, (4) severe focal injury tat may include necrosis or irreversible loss of function, and (5) overwhelming and rapidly fatal diffuse injury associated with high-dose, whole-body exposures. An understanding of the implications of nervous system exposure to radiation can guide efforts in radiation protection and aid in the optimization of the medical uses of radiation. PMID- 15757799 TI - Neurobiological weapons. AB - Biological warfare is a potential threat on the battlefield and in daily life. It is vital for neurologists and other health care practitioners to be familiar with biological and toxic agents that target the nervous system. most illnesses caused by biological warfare agents are not commonly considered neurologic disease, however. Many of these agents (such as anthrax) may present with headache, meningitis, or mental status changes in addition to fever and other symptoms and signs (Tables 2 and 3). Thus, a neurologist may be consulted acutely to aid in diagnosis. Because of the incubation time of many biological agents and their protean manifestations, it is likely that health care workers will be on the front lines in the event of a bioterrorist attack. We must be prepared. PMID- 15757800 TI - Nerve agents. AB - Nerve agents cause a rapidly fatal cholinergic crisis, but rapid, appropriate antidotal treatment saves lives. Survivors of nerve-agent poisoning generally are healthy, unlike survivors of some other chemical agent attacks. Neurologists can assist first responders and mass casualty planners materially by serving as resources for information on nerve agents and the syndromes they cause. They also can help their communities by reinforcing that treatment for nerve-agent poisoning is effective. PMID- 15757802 TI - Epidemiology and natural history of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is a major contributor to cancer incidence and mortality. There is a wide variation, however, in the global distribution of HCC. Eighty percent of the burden is borne by countries in Asia and sub-Saharan Africa. In most high-risk countries, principal risk factors include infection with hepatitis B virus and dietary exposure to aflatoxin B(1). In contrast, hepatitis C virus and alcohol consumption are more important risk factors in low risk countries. In recent years, the incidence of HCC has decreased in some high risk countries and increased in some low-risk countries. Reasons for both trends are not completely understood, but are likely related to public health efforts in Asia and the increase in hepatitis C virus infection in low-risk countries. Vaccination programs against hepatitis B virus will likely decrease the HCC rate even further in decades to come. PMID- 15757803 TI - Molecular mechanisms in hepatocellular carcinoma development. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers in the world. It usually develops in patients with chronic viral hepatitis, aflatoxin exposure, or excessive alcohol use. Most patients with HCC present with advanced disease and have a poor prognosis. The implementation of antiviral drugs and the availability of a vaccine for hepatitis B should help reduce the incidence of HCC. Considerable effort has now focused on unraveling the molecular pathogenesis of HCC in order to design better treatments, or to prevent the disease altogether. However, so far, the pathogenesis of HCC appears to be quite heterogeneous among patients. In particular, several mechanisms of tumorigenesis seem to be involved, including loss of tumor suppressor gene function, oncogene activation, direct viral effects, DNA methylation, and angiogenesis. It is not clear which events are critical in tumor initiation versus tumor progression. RNA expression arrays and proteomics hold promise to provide further clues about this common and complex cancer. PMID- 15757804 TI - Histopathology of liver cancers. AB - Recently, new pathomorphologic information about early-stage small hepatocellular carcinoma (HCC) and the multi-step process of human hepatocarcinogenesis has been obtained, along with advances in the development of diagnostic modalities. The most valuable information is that in the majority of cases HCC arises as a very well differentiated cancer and proliferates with a stepwise process of dedifferentiation. In addition, it has been suggested that many HCCs seem to arise from dysplastic nodules (DNs) on the basis of the following evidence: the presence of DNs containing HCC foci, frequent association of DNs in the vicinity of HCC, and clinical progression from DN to HCC. However, as many HCCs are still detected at an advanced stage, it is also important to understand not only the classical pathologic features of HCC but also unusual features such as scirrhous change, sarcomatous change, fibrolamellar variant, and intra-bile duct or intra atrial tumor growth. PMID- 15757805 TI - Multidetector CT of hepatocellular carcinoma. AB - Hepatocellular carcinoma is one of the most common causes of cancer death worldwide. Numerous surgical and nonsurgical treatment options are available for the management of patients with HCC. Successful long-term outcome is dependent on early detection of HCC, as well as accurate delineation of the number and location of tumor nodules. We present the different manifestations of hepatocellular carcinoma as depicted by multidetector-row CT with advanced image processing. PMID- 15757806 TI - Serological markers of liver cancer. AB - Serological markers for hepatocellular carcinoma (HCC) are important for early diagnosis, as well as monitoring of tumour aggressiveness, treatment responsiveness, recurrence and survival. The three most common markers are total alpha-fetoprotein (AFP), Lens culinaris agglutinin-reactive AFP (AFP-L3) and protein induced by vitamin K absence or antagonist-II (PIVKA-II). Total AFP has the sensitivity of 60% and specificity of 90% for the detection of HCC. Increase in the percentage of AFP-L3 over the total AFP (>10%) is very specific for small HCC. PIVKA-II is also more specific than total AFP in detecting HCC. AFP-L3 and PIVKA-II levels correlate with tumour aggressiveness and prognosis. All three markers are useful for monitoring treatment responsiveness and tumour recurrence. Since the levels of the three markers are independent of each other, combination of measurement of two or three markers will increase the sensitivity and diagnostic accuracy. Some novel markers including glypican-3 are being extensively studied. PMID- 15757807 TI - Screening for hepatocellular carcinoma. AB - There is currently no evidence that screening patients at risk for hepatocellular carcinoma reduces mortality from the disease. Nonetheless, screening is widely practiced. Screening is a process that includes selecting patients, applying screening tests, deciding on recall policies, and subsequently proving or disproving the presence of cancer. The literature on screening for hepatocellular carcinoma is confusing at best, and does not adequately consider the many biases that result from uncontrolled and retrospective studies. Nonetheless, screening can be justified because it is likely that mortality is decreased by adequate treatment of small cancers, particularly in the era of liver transplantation. False-positive screening test results are common. Once an abnormal screening result is obtained there is little guidance from the literature as to how patients should be investigated further, nor about how to determine whether the screening test result was a false-positive. This should at minimum include short interval follow-up with CT scans and MRI's. PMID- 15757808 TI - Clinical trial design in hepatocellular carcinoma. AB - Researchers interested in developing treatments for hepatocellular carcinoma (HCC) face three sets of problems; one set common to all clinical trialists at the beginning of the 21st century, one set related to phase I/II studies ('early drug development') and one set related to the design of phase III (randomised, controlled trials). This review is concerned with the challenges faced in designing high quality clinical trials in patients with HCC. Specific published trials are not discussed other than by way of illustration of the problems faced. PMID- 15757809 TI - Treatment of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common malignant tumours worldwide. The major etiologies and risk factors for HCC development are well defined and some of the multiple steps involved in hepatocarcinogenesis have been elucidated in recent years. Despite these scientific advances and the implementation of measures for early HCC detection in patients at risk, patient survival has not improved during the last three decades. This is due in part to the advanced stage of the disease at the time of clinical presentation, in part due to the limited therapeutic options. These fall into four main categories: (1) surgical interventions, including tumour resection and liver transplantation, (2) percutaneous interventions, including ethanol injection and radiofrequency thermal ablation, (3) transarterial interventions, including embolisation and chemoembolisation and (4) drugs as well as gene and immune therapies. These therapeutic strategies have been evaluated in part in randomised controlled clinical trials that are the basis for therapeutic recommendations. While surgery and percutaneous as well as transarterial interventions are effective in patients with limited disease (1-3 lesions, < 5 cm in diameter) and compensated underlying liver disease (cirrhosis Child A), at the time of diagnosis more than 80% patients present with multicentric HCC and advanced liver disease or comorbidities that restrict the therapeutic measures to best supportive care. In order to reduce morbidity and mortality from HCC, therefore, early diagnosis and the development of novel systemic therapies for advanced disease, including drugs, gene and immune therapies as well as primary HCC prevention are of paramount importance. Further, secondary HCC prevention after successful therapeutic interventions needs to be improved in order to make an impact on the survival of patients with HCC. New technologies, including gene expression profiling and proteomic analyses, should further elucidate the molecular events underlying HCC development and identify novel diagnostic markers as well as therapeutic and preventive targets. PMID- 15757810 TI - Liver transplantation for hepatocellular carcinoma. AB - In patients with cirrhosis and hepatocellular carcinoma (HCC), orthotopic liver transplantation (OLT) offers hope for cure of both the complicating HCC and the underlying chronic liver disease. Excellent 5 year survival has been reported when the restrictive Milan criteria are used to select transplant candidates. Alternative recommendations have recently been proposed by groups at University of California San Francisco, University of Pittsburgh and Mount Sinai. We review current and evolving concepts regarding selection criteria for OLT in patients with HCC, along with strategies to reduce waiting times, such as the impact of the implementation of the model for end-stage liver disease (MELD) scoring system on organ distribution and the role of living donor OLT for this indication. The possible efficacy of adjuvant anti-tumour therapies in limiting HCC growth while waiting for OLT, along with factors influencing the risk of HCC recurrence post OLT, the major cause of death in this setting, are also discussed. PMID- 15757811 TI - General management. AB - Hepatocellular carcinoma is one of the most common malignancies worldwide. The general management of hepatocellular carcinoma begins with an accurate diagnosis. With advances in imaging studies, noninvasive diagnosis has become an accepted standard of care for hepatocellular carcinoma, though pathologic examination is still required in selected cases. Following diagnosis, accurate staging is the next most important step in selecting the most appropriate treatment modality. Patients with localised tumor and compensated liver disease should be considered for partial hepatectomy, and patients with poor hepatic function but early tumor stage are candidates for liver transplantation. Patients who do not qualify for either of these curative treatments may be evaluated for palliative therapy, of which transarterial chemoembolisation is most widely used. This review will discuss the role of biopsy, the pros and cons of noninvasive and pathologic tissue diagnosis as well as the general approach to choose the most appropriate treatment for patients with hepatocellular carcinoma. PMID- 15757812 TI - The skin immune system and the challenge of tumour immunosurveillance. AB - The Skin Immune System (SIS) is a relatively new concept central to the issue of cutaneous tumour surveillance. The Langerhans cell (LC) is a key component of SIS. Skin cancer causing agents such as ultraviolet B (UV-B) irradiation and chemical carcinogens like dimethylbenz(a)anthracene (DMBA) alter LC function, resulting in immunosuppression and the promotional phase of tumour development. Once tumours, such as melanoma, are established they may show evidence of tumour regression due to immune reaction but frequently escape immune attack and metastasise. This article explores our knowledge of LC and SIS in these responses. For tumour immunosurveillance to be an effective reality at the clinical level, experiments are required to provide a more precise base for immunotherapy. PMID- 15757813 TI - Familial occurrence of axillary papular mucinosis. AB - Primary cutaneous mucinosis encompasses a wide range of different skin conditions with circumscribed, follicular or diffuse manifestation. For the first time, to our knowledge, we report on the occurrence of so-called "atypical" localized papular mucinosis in the axillary pits of a mother and her (identical) twin daughters presenting as asymptomatic soft minuscule papules. This unique observation allows us to define an autosomal dominant inheritance pattern with a high level of penetrance for this rare skin disease. PMID- 15757814 TI - Congenital blaschkoid eccrine spiradenoma on the face. AB - Eccrine spiradenoma is an uncommon tumour of the sweat gland(s) and generally occurs as a solitary small painful nodule. Very rarely, it may present as multiple tumours arranged in a linear/blaschkoid pattern. An 8-year-old girl had many papulonodular lesions, arranged in a linear pattern on the left half of the face, involving the forehead and lateral side of the nose. The lesions were along Blaschko's lines. Lesions were reported to be present at birth; no family member had similar lesion(s), either in solitary or multiple form. Histopathologic examination of a biopsy specimen revealed characteristic features of eccrine spiradenoma. The case is reported because of the rare occurrence of this presentation. PMID- 15757815 TI - Bigenic connexin mutations in a patient with hidrotic ectodermal dysplasia. AB - Gap junctions are formed by a polygenic family of more than 20 different connexin proteins. They mediate intercellular communication via the direct exchange of ions, metabolites and secondary messengers, thus controlling and coordinating cellular activities. Mutations in five gap junction genes, including GJB2 (Cx26), GJB3 (Cx31), GJB4 (Cx30.3), GJB6 (Cx30) and GJA1 (Cx43) are known to cause inherited hearing loss and/or disorders of the skin and its appendages, often giving rise to overlapping phenotypes. In this study we present a patient with hidrotic ectodermal dysplasia, who had abortive features of oculo-dento-digital dysplasia, extensive hyperkeratosis of the skin. The patient harbored a novel sporadic mutation (V41L) in GJA1 (Cx43) as well as a heterozygous coding variant (R127H) of GJB2 (Cx26). Our findings suggest that GJA1 mutations can produce variable clinical phenotypes on the background of sequence variants in other connexins. PMID- 15757816 TI - Desmosome-binding antibody KM48 recognises an extracellular antigen different from desmosomal cadherins Dsg 1-3 and Dsc 1-3. AB - Desmosomes are the most prominent and mechanically important epidermal intercellular junctions. Transmembrane proteins of desmosomes, desmogleins and desmocollins, are responsible for extracellular binding and, thus, are important for interkeratinocyte cohesion. We show here, using three different approaches, that the extracellular "cores" of epidermal desmosomes contain a highly glycosylated antigen, different from desmosomal cadherins. This protein, recognised by KM48 monoclonal antibody, is likely to be involved in the processes of keratinocyte differentiation, desmosome turnover and epidermal cohesion. PMID- 15757817 TI - Comparison of topical paromomycin sulfate (twice/day) with intralesional meglumine antimoniate for the treatment of cutaneous leishmaniasis caused by L. major. AB - This is an open study to compare the cure rate of cutaneous leishmaniasis caused by L. major and treated with either paromomycin sulfate or intralesional injection of meglumine antimoniate. Sixty parasitologically proven cases with 1-3 lesions were included and divided randomly into two equal groups; one group received 1 ml of meglumine antimonate intradermally every other day for 20 days, the other group received the ointment containing 15% parmomycin sulfate in urea twice daily for 20 days. The patients were clinically evaluated at 1 and 6 weeks after treatment was completed. The results of clinical evaluation at 1 week after treatment completed showed a cure rate of 18 out of 27 (66%) in the meglumine antimonate injected group and 20 out of 29 (68%) in the paromomycin sulfate treated group. The chi square test was used to compare the cure rate between the two groups and showed no significant difference (p = 0.85). PMID- 15757819 TI - Treatment of congenital nevi with the Q-switched Alexandrite laser. AB - Q-switched mode lasers have been utilized for treatment of many pigmentary lesions. Because of their short pulse durations (1-100 ns), these lasers selectively destroy pigment laden cells while sparing the surrounding normal tissues. To determine if the Q-switched alexandrite laser (QSAL) is effective in the treatment of congenital melanocytic nevi, 53 patients with these lesions were treated with QSAL; of these, 16 also received CO(2) laser treatment between QSAL treatments. We observed an average improvement score in the 53 treated patients of 2.623 + 1.13, corresponding to a 72% improvement. Treatment with the combination of QSAL and a CO(2) laser resulted in a significant enhancement of improvement score (3.06 +/- 1.18) compared to patients treated with the QSAL alone (2.43 +/- 1.07; p = 0.0393). Complications were mild, with 35 nevi (67.3%) showing textural change of skin after treatment, 2 (3.8%) showing depressed scar formation, and 4 (7.5%) showing hypertrophic changes, while 12 nevi (23%) showed no changes. Hypopigmentation was observed in 16 patients (30%), and hyperpigmentation was observed in 15 patients (28%) 48 weeks after the final QSAL treatment. Repigmentation to a brown to black spot was observed in 44/53 (83%) patients within an average of 5.45 +/- 3.93 months. These results indicate that the QSAL was as effective as other Q switch mode lasers in the treatment of congenital melanocytic nevi, but repigmentation is a problem. PMID- 15757818 TI - Topical 0.05% clobetasol propionate versus 1% pimecrolimus ointment in vitiligo. AB - Vitiligo is a common skin condition resulting from loss of normal melanin pigments in the skin which produces white patches. Topical corticosteroids are indicated for the treatment of limited areas of vitiligo. Pimecrolimus, which inhibits calcineurin, has recently been shown to be effective for the treatment of vitiligo. We performed a prospective study to evaluate the efficacy of the 0.05% clobetasol propionate and 1% pimecrolimus in the treatment of vitiligo. Ten patients with virtually bilateral symmetrical lesions of vitiligo were included. 0.05% clobetasol propionate was applied twice daily over the lesion on right side of the body, and topical 1% pimecrolimus was applied twice daily over the lesion on left side of the body. It was determined that both treatment modalities resulted in a comparable rate of repigmentation. Response to treatment was varied according to the anatomical location of the lesions where better results were seen on the trunk and extremities. Results from this pilot study indicate that topical 1% pimecrolimus is as effective as clobetasol propionate in restoring skin disfiguring due to vitiligo. For a better conclusive statement further studies involving larger groups of patients should be performed. PMID- 15757820 TI - Maculo-papular rash induced by lopinavir/ritonavir. AB - Two patients with HIV disease developed a pruritic, maculo-papular rash shortly after introducing an association of 2 protease inhibitors (PI) lopinavir/ritonavir (Kaletra). The dermatitis started respectively 7 and 10 days after taking Kaletra, improved on withdrawal, and relapsed following its reintroduction. Although itchy, the dermatitis was not life-threatening, and the patients were treated "through the rash" without suspending the drug. Histopathologic examination of lesional skin showed a non-specific inflammatory infiltration consisting of neutrophils and lymphocytes, but there were no eosinophils, nor dilatation of capillaries in the papillary dermis. HIV positive patients are more prone to drug-related rashes than the general population. Awareness of the side-effects of PI plays an important role in keeping compliance with the treatment and helps patients reach their goal in controlling the development of HIV. The safety profile of Kaletra and its cutaneous side effects have yet to be completely elucidated. PMID- 15757821 TI - The nevoid pili multigemini over the back. AB - Pili multigemini is a rare developmental defect of hair follicles. It usually involves the beard of an adult and the scalp in children, but there have been no reported cases on the back. We report 3 cases of extensive pili multigemini over the entire back of healthy males. All cases were found incidentally. The direct microscopic examination of one hair follicle showed that it consisted of more than twenty shafts, which had rough surfaces and brush-like bulbs. The histological examination revealed that complicated follicular structures formed as many as 23 to even 36 hair shafts. Lots of hairs were surrounded by each inner root sheath and had one common outer root sheath. We suggest this nevoid is pili multigemini over the back. PMID- 15757822 TI - Regression of cutis calcinosis with diltiazem in adult dermatomyositis. AB - Calcinosis cutis is common in several connective tissue diseases such as dermatomyositis, scleroderma or lupus erythematous. In dermatomyositis, it is more likely to concern children than adults but it is not exceptional in adults. Several treatments have been used empirically with inconsistent success. We report a case of adult cutis calcinosis associated with dermatomyositis which responded dramatically to treatment with diltiazem. PMID- 15757823 TI - Folliculosebaceous cystic hamartoma with hair shaft fragments. AB - We describe herein a 57-year-old Japanese male with a folliculosebaceous cystic hamartoma (FCH) presenting several hair shaft fragments in the cyst. Clinically, a skin-colored, dome-shaped, hard nodule, 5 x 4 mm in diameter was observed on the antitragus of his left ear. Histological examination revealed a large cystic structure surrounded by multiple mature sebaceous lobules and rudimentary hair follicules in the middle of the dermis. The cystic structure contained laminated orthokeratotic keratin and several hair shaft fragments. The fibrous stroma invaded the sebaceous lobules and obvious clefts were seen around the folliculosebaceous units located in the dermis. These features are characteristic histological findings of FCH. To our knowledge, this case is unusual with the presence of hair shaft fragments within the cystic structure. In addition, we discuss the histopathological findings of FCH and also review other cases of FCH in the literature. PMID- 15757824 TI - Extensive psoriasis induced by pegylated interferon alpha-2b treatment for chronic hepatitis B. AB - A 32-year-old man with chronic hepatitis B who was treated with pegylated interferon alpha-2b once a week, developed widespread psoriasis 4 weeks after the beginning of the treatment. There was no history of psoriasis. The treatment was stopped and gradually the eruption vanished. Thereafter, the patient was treated with lamivudine successfully without dermatological or other sequelae. PMID- 15757826 TI - Melanoma of the glans penis. AB - The authors describe the case of a 64-year old man who presented with an asymptomatic brown macula on his glans penis that had appeared about 18 months earlier. Dermoscopy analysis demonstrated a prominent, wide and irregular pigment network, which stopped abruptly at the periphery of the lesion. A diagnostic biopsy showed the characteristics of a melanoma in situ . The patient was referred for partial surgical excision of the glans. No recurrence or metastasis occurred during the two years after the operation. Melanoma in situ of the penis is very rare in dermatologic literature. Early diagnosis is of paramount importance because its prognosis is very poor. Early systematic use of dermoscopy may be useful for the differential diagnosis of pigmented mucosal lesions, which include mucosal melanosis and other benign melanoses of genitalia. PMID- 15757825 TI - Necrolytic migratory erythema with myelodysplastic syndrome without glucagonoma. AB - Necrolytic migratory erythema is a cutaneous paraneoplastic manifestation, which is usually associated with a glucagon-secreting pancreatic tumor. However, it also may occur in other circumstances in which serum glucagon is elevated, as in hepatic cirrhosis. Rarely, necrolytic migratory erythema is reported in association with a jejunal and rectal adenocarcinoma or villous atrophy of the small intestine without any evidence for increased serum glucagon levels. In this context we report the case of an 85-year-old male with myelodysplastic syndrome who developed typical necrolytic migratory erythema without glucagonoma syndrome or evidence for other pancreatic or liver disease. We suggest that, in addition to the diseases listed, myelodysplastic syndrome might be able to cause necrolytic migratory erythema. PMID- 15757827 TI - The Southeast Asian 12.5 KB (delta-beta) degrees-deletion: a common beta thalassemia in Mon-Khmer groups (Lao Theung) of South Laos. AB - Seven patients with unexplained anemia and mild thalassemic features were ascertained during a survey of hemoglobinopathies in the Sekong Province in South Laos. These patients belong to the Austroasiatic (Mon-Khmer) population of South Laos (official designation Lao Theung). Hemoglobin electrophoresis on cellulose acetate showed absence of Hb A and two bands in the positions of Hb E and Hb F respectively. Sequencing of DNA isolated from venous blood revealed the codon 26 G-->A mutation characteristic of the HBB*E gene, but none of the common Southeast Asian beta-thalassemia mutations were found. Detailed studies in four of the seven subjects identified a 12.5 kb deletion encompassing part of the delta globin gene and the entire beta-globin gene. We conclude that this deletion is a common, and possibly the predominant beta-thalassemia mutation of the Austroasiatic Lao Theung population. Similar deletions reported in single individuals in Laos, Thailand and Vietnam are probably due to migrational spreading to areas adjacent to South Laos. PMID- 15757828 TI - Influence of body morphology on turning behavior in carnivores. AB - The present work illustrates how three carnivore species with different body morphologies differ in the performance of a basic motor component-turning. Marbled polecats, which have an elongated and slender trunk, turn while sharply bending the trunk laterally. Grey wolves possess elongated legs and turn by maneuvering with their legs while slightly bending the trunk laterally and lowering the head to contact the ground. Honey badgers feature a wide and massive bear-like shape and rarely bend the trunk, but rather turn either by maneuvering with their legs or while elevating parts of the trunk in the vertical domain. It is suggested that these strategies shorten the radius of turning and thus reduce the moment of inertia. A lower moment of inertia may optimize turning behavior in terms of speed, energy cost, and the smoothness of transition between turning and bouts of forward progression or arrest. PMID- 15757829 TI - The interaction of response complexity and instructional set. AB - This study investigated the interaction of experimental instructions and response complexity in an attempt to provide a possible explanation for some of the equivocalness in the programming time literature. Response complexity was manipulated by varying the accuracy required of subjects striking circular targets in a simple reaction time paradigm. Subjects were tested on two days, one week apart, during which experimental instructions emphasized either leaving the start position as soon as possible (initiation emphasis) or completing the response with a rapid but smooth movement (form emphasis). Subjects had shorter reaction times but longer movement times when they performed under initiation emphasis instructions rather than under form-emphasis instructions. Furthermore, in contrast to form-emphasis conditions, there was no effect of response complexity on reaction time in conditions of initiation emphasis. It appears that subtle changes in experimental instructions can lead to very different patterns of reaction time and movement time data. These findings highlight the need for caution in preparing subject instructions in response programming experiments. PMID- 15757830 TI - On the cognitive processes underlying contextual interference and observational learning. AB - The main goal of the present study was to determine whether observation of an unskilled model learning a timing task enables the observer to develop a cognitive representation of the task similar to the one acquired through physical practice (Adams, 1986; Bandura, 1977; Lee & White, 1990). To reach that goal, we tested whether a contextual interference effect would be obtained in a retention test of subjects who had observed an individual practicing three variations of a timing task under a random or a blocked schedule of practice. Similar patterns of results in an immediate retention test were found following observation and physical practice. This suggests that observation indeed engaged the observers in the same type of cognitive activities as did physical practice. Moreover, a schedule of practice made up of 100% physical practice led to improved learning compared with a schedule of practice made up of 50% observation followed by 50% physical practice. This suggests that learning is enhanced more by numerous implementations of a motor program than by its mere construction or retrieval. PMID- 15757831 TI - Resolving a conflict between sensory feedback and knowledge of results, while learning a motor skill. AB - In a recent study, Buekers, Magill, and Hall (1992) showed that even when verbal knowledge of results (KR) was redundant with sensory feedback, erroneous KR influenced the learning of motor skills. To determine why this occurred, we conducted two experiments. In Experiment 1, subjects performed 50 practice trials on a complex anticipation task and then performed three non-KR retention tests of 25 trials each. The results indicated that when correct KR and erroneous KR were provided alternately, subjects ignored the erroneous KR and performed according to the correct KR. Experiment 2 compared different ratios of no KR to erroneous KR. The results showed that, for low ratios (1:1 and 4:1), learning experience was similar to a condition in which erroneous KR was presented on all trials. For a higher ratio (9:1), however, learning performance was similar to performance when KR was correct on all trials or was not presented. These results are interpreted as support for the hypothesis that when two conflicting sources of information are available, the subject's degree of uncertainty about the valid source of information influences his selection of the information to guide performance. PMID- 15757832 TI - Resolving power of the perceptual- and sensorimotor systems in 6- to 10-year-old children. AB - Children aged 6 to 10 were tested on their ability to move accurately and to perceptually evaluate their motor response. Subjects performed a directional and an amplitude visuo-manual aiming task without vision of their moving limb. They were asked to correct their error, after completion of their movements, only if they felt they were not accurate. Terminal aiming errors and correction responses (adjustments) were analyzed, and threshold detection was determined relative to terminal aiming error. Action accuracy and evaluation of action accuracy are two abilities that do not develop synchronously. Moreover, the relationship between these abilities depends on whether accuracy and direction or amplitude are required. Amplitude undergoes more corrections than direction, suggesting that the spatial system of reference involved depends more upon the coding of the final position than on direction. Two spatial comparators, operating on the basis of two types of evaluation, seem to have a variably distinct contribution to movement and perception accuracy, according to age. PMID- 15757833 TI - The effects of temporal-precision and time-minimization constraints on the spatial and temporal accuracy of aimed hand movements. AB - Discrete aimed hand movements, made by subjects given temporal-accuracy and time minimization task instructions, were compared. Movements in the temporal-accuracy task were made to a point target with a goal movement time of 400 ms. A circular target then was manufactured that incorporated the measured spatial errors from the temporal-accuracy task, and subjects attempted to contact the target with a minimum movement time and without missing the circular target (time-minimization task instructions). This procedure resulted in equal movement amplitude and approximately equal spatial accuracy for the two task instructions. Movements under the time-minimization instructions were completed rapidly (M = 307 ms) without target misses, and tended to be made up of two submovements. In contrast, movements under temporal-accuracy instructions were made more slowly (M = 397 ms), matching the goal movement time, and were typically characterized by a single submovement. These data support the hypothesis that movement times, at a fixed movement amplitude versus target width ratio, decrease as the number of submovements increases, and that movements produced under temporal-accuracy and time-minimization have different control characteristics. These control differences are related to the linear and logarithmic speed-accuracy relations observed for temporal-accuracy and time-minimization tasks, respectively. PMID- 15757834 TI - A note on data smoothing for movement analysis: the relevance of a nonlinear method. AB - The goal of this experiment was to test a potentially useful nonlinear method for smoothing noisy position data, which often is encountered in the analysis of data. This algorithm (7RY) uses a nonlinear smoothing function and behaves like a low-pass filter, automatically removing aberrant points; it is used prior to differentiation of time series so that usable acceleration information can be obtained. The experimental procedure comprises position data collection along with direct accelerometric data recording. From the position-time data, (a) 7RY and (b) Butterworth algorithms have been used to compute twice-differentiated acceleration curves. The directly recorded acceleration measurements were then compared with the acceleration computed from the original position data. Although the results indicated an overall good fit between the recorded and the calculated acceleration curves, only the nonlinear method led to reliable acceleration curves when aberrant points were present in the position data. PMID- 15757835 TI - A new book by N. A. Bernstein: "On dexterity and its development". AB - A book, "On Dexterity and Its Development," written by N. A. Bernstein in the 1940s has recently been published in Moscow. We describe briefly the amazing fate of this miraculously saved book and of its famous author. The book is unique in its reader-friendly handling of the most complicated issues of motor control, motor development, motor skill acquisition, and more general aspects of brain activity. It presents the basis for what is now called motor control and for some of the more fundamental issues of theoretical neuroscience. Bernstein described his views on multilevel control of movements, the role of sensory feedback, preprogramming, feedback and feedforward components in motor control, the role of exercise, and other subjects. Despite the fact that the book was written nearly half a century ago, its deep philosophical content remains beyond any competition. PMID- 15757836 TI - The relative dominance of schemata in a manual tracking task: input patterns, system dynamics, and movement patterns. AB - As subjects repeatedly track the same spatiotemporal pattern, they learn to anticipate the pattern and thereby to reduce their effective time delay. The successive organization of perception hypothesis suggests that subjects progress from an error-nulling mode to a pursuit mode, in which they respond to the estimated instantaneous input pattern, and then to a precognitive mode, in which they rely on memory of the input pattern or movement pattern. We used a transfer of training experiment involving position and rate control systems to distinguish among these strategies by determining the relative dominance of schemata for the input pattern, system dynamics, and movement pattern. Results indicated that subjects relied on a memorized movement pattern, which is characteristic of the precognitive mode. Adaptations of this transfer methodology may be useful in testing other theories that postulate multiple schemata for the control of perceptual-motor behavior. PMID- 15757837 TI - Eye-hand coordination: spatial localization after saccadic and pursuit eye movements. AB - Subjects produced speeded and unspeeded hand movements to a target location after either saccadic or pursuit eye movements to the target. Hand movements began either aligned with the initial position of gaze or from some other location. Subjects generally underestimated the extent of the pursuit eye movements relative to estimates made after saccades. With speeded hand movements, however, the underestimation was reduced considerably if the hand movements began aligned with a location other than the initial position of gaze. The results reveal details of the mechanisms underlying eye-hand coordination and show that important differences exist in the information used for localization for slow and rapid limb movements. PMID- 15757838 TI - Effects of optic flow on the kinematics of human gait: a comparison of young and older adults. AB - This experiment studied the effect of imposed optic flow on human locomotion. Six young and 6 older adults were exposed to various patterns of optic flow while walking in a moving hallway. Results showed few cases of impaired postural control (staggers, parachute reactions). No falls were recorded. Kinematic patterns of gait were altered when vision was absent or inconsistent optic flow was presented: Ninety two percent of the subjects' mean step velocity differed from their step velocities under normal vision. Compared with imposed central flow, peripheral optic flow was not dominant in inducing kinematic changes. Characteristic gait profiles were obtained, depending on flow direction. Global backward flow tended to slow down step velocity, whereas subjects' step velocity increased during conditions of forward flow. The results suggest that subjects attempted to match their own walking speed to the velocity of the moving visual scenes. It is concluded that in an uncluttered environment, imposed optic flow has a modulating rather than a destabilizing effect on human locomotion. PMID- 15757839 TI - The effects of environmental changes on one-handed catching. AB - Ball catching involves predicting the time and place of arrival of a mobile object. Visual cues of various kinds may help a ball catcher to perform this task successfully. The aims of the present study were (a) to assess the role of the environment in the spatiotemporal planning of ball-catching movements and (b) to determine what specific cues are actually used for this purpose. In the first experiment described here, subjects' catching performances were compared under four different environmental conditions, namely, normal lighting, ultraviolet light with no background, ultraviolet light with a densely structured background, and ultraviolet light with a sparsely structured background. Our results showed that the sight of the mobile object alone does not provide enough information for a subject to achieve his maximum performance level. Accurately assessing the point of arrival of the ball requires the use of relative visual cues. The environment is also a source of visual cues used to assess the time of arrival of the ball. A second experiment was carried out with a view to determining the exact nature of the visual cues used. Here, the orientation (frontal vs. oblique plane) and the apparent visual angle (6 degrees vs. 42 degrees ) of the background were made to vary. The results of this experiment showed that the orientation of the background affected the percentage of spatial errors produced by the subjects, whereas the apparent visual angle affected the percentage of temporal errors. The relative velocity cue generated by the masking of successive structures in the environment by the oncoming ball seems to have been taking into account in estimating the time of arrival of the ball. This cue seems to be of crucial importance during the 200 ms prior to the time of contact between the ball and the subject's hand. This finding supports the idea that the method used to assess the time to contact may involve velocity information. PMID- 15757840 TI - Effect of a terminal constraint on control of balance during sit-to-stand. AB - The speed at which sit-to-stand (STS) motions are performed and the subsequent terminal constraint upon upright stance can present subjects with contradictory goals. Previous findings suggested that subjects might adopt a strategy of limiting the peak horizontal momentum of the center of mass (CM), and perhaps of body segments as well, regardless of the speed of ascent. The primary purpose of this study was to test the hypothesis that the limitation in CM momentum is related to the constraint on upright stance at the termination of the task. The secondary purpose was to describe the contribution of the shank, thigh, and upper body (head-arm-trunk) to the peak horizontal momentum of the CM under each test condition. Nine healthy adult males rose from a seated position under the following three conditions: (a) at natural speeds (natural STS); (b) as fast as possible (fast STS); and (c) as fast as possible, followed by falling forward while keeping the feet fixed and using the arms on a support bar to stop the fall (fast STS+fall). The results showed that the peak horizontal momentum of the CM did not change substantially from the natural to fast STS, but increased significantly from the fast STS to the fast STS+fall. These findings are consistent with the hypothesis that limiting peak horizontal momentum of the CM may reflect a movement control strategy related to maintaining equilibrium at the termination of the voluntary task of rising from a chair. The momentum profile of the upper body, but not of the thigh or shank, remained constant across all experimental conditions, suggesting that the motion of the upper body may be tightly controlled in STS, regardless of the altered constraint on balance. PMID- 15757841 TI - Effects of movement duration and visual feedback on visual and proprioceptive components of prism adaptation. AB - While looking through laterally displacing prisms, subjects pointed sagittally 80 times at an objectively straight-ahead target, completing a reciprocal out-and back pointing movement ever 1, 3, or 6 s. Visual feedback was available early in the pointing movement or only late at the end of the movement. Aftereffect measures of adaptive shift (obtained after every 10 pointing trials) showed adaptive change only in limb position sense (i.e., proprioceptive adaptation) when movement duration was 1 s, regardless of visual feedback condition; but as movement duration increased, adaptive change in the eye position sense (i.e., visual adaptation) increased while proprioceptive adaptation decreased, especially for the late visual feedback condition. Regardless of visual feedback condition, proprioceptive adaptation showed the maximal rate of growth with the 1 s movement duration, whereas visual adaptation showed maximal growth with the 6-s movement duration. These results provide additional support for a model of adaptive spatial mapping in which the direction of strategically flexible coordination (guidance) between eye and limb (and consequently the locus of adaptive spatial mapping) is jointly determined by movement duration and timing of visual feedback. An additional effect of movement duration is to determine the rate of discordant inputs. Maximal growth of adaptation occurs when the input rate matches the response time of the spatial mapping function. PMID- 15757842 TI - Combined effects of movement velocity and duration on programming time: Spijkers (1989) revisited. AB - The goal of the present study was to determine the combined effects of movement velocity and duration on motor programming. Subjects were submitted to a two choice reaction time task that could be completed by aiming movements differing in the mean velocity at which they were to be produced as well as by their movement time. The results of the present study indicate that, in each pair of responses used, the responses having the higher mean velocity were initiated faster that those having the lower mean velocity. Contrary to Spijkers' (1989) study, the different movement time pairings did not modify the effect of movement velocity on response programming time. Moreover, the same pattern of results was observed whether or not the subjects were permitted to visually guide their ongoing movement. Thus, Spijkers' proposition, that the type of control one may use to guide an aiming movement needs to be determined before movement initiation can take place, was not confirmed. PMID- 15757843 TI - Motor skill acquisition and retention as a function of average feedback, summary feedback, and performance variability. AB - Summary feedback involves withholding feedback from subjects until the last trial in a block is completed, and then presenting feedback about each trial. A variation of this method, called average feedback (Young & Schmidt, 1992), presents subjects with only the mean of the trial block. We investigated whether these methods have similar effects on acquisition and retention of a simple motor skill. Five groups of subjects (n = 16 per group) performed 60 acquisition trials of an aiming task involving both spatial and temporal accuracy. We presented average and summary feedback based on either 5-trial blocks or 15-trial blocks and compared these schedules with every-trial feedback. During acquisition, all groups improved with practice, with a slight tendency for the every-trial condition to have less absolute error than the longer summary and average conditions. Analysis of delayed no-feedback retention tests, however, revealed a strong advantage for the 5-trial summary and average conditions compared with the every-trial condition. In addition, we found that for long blocks of acquisition trials without augmented feedback, the performance variability of those trials was associated with retention performance. Results are discussed in terms of how these different manipulations may make feedback less useful during acquisition, but foster the use of certain information processing activities that enhance overall learning. PMID- 15757844 TI - Partial visual feedback and spatial end-point accuracy of discrete aiming movements. AB - Five experiments are reported in which the effect of partial visual feedback on the accuracy of discrete target aiming was investigated. Visual feedback was manipulated through a spectacle-mounted liquid-crystal tachistoscope. The length of the visual feedback interval was varied as a percentage of the instructed movement time. In Experiment 1, the length of the vision interval was manipulated symmetrically at the beginning- and end-phase of the movement, whereas in the remaining experiments, the vision time was varied with respect to the end-phase only. The variations at the end were examined for different distances (Experiment 2), different movement speeds at the same distance (Experiment 3), and in small interstep intervals (Experiment 4). A vision time of more than 150 ms at the end phase of the movement enhanced aiming performance in all experiments. Longer vision times monotonously improved aiming accuracy; the fifth experiment showed that a vision time of about 275 ms was sufficient for near-perfect aiming. Furthermore, the significance of vision during the first phase of a movement was demonstrated again. The results of the five experiments pointed to shorter visuomotor processing times. To explain the beneficial effects of short vision times for aiming accuracy, we propose a model of visuomotor processing that is based on the stochastic optimized submovement model of Meyer, Abrams, Kornblum, Wright, and Smith (1988). PMID- 15757845 TI - Idiosyncratic errors in visually directed reaching. AB - Subjects reaching with their eyes closed to visually presented targets missed their targets by an average of 4% of arm length; significant variation in the direction and magnitude of error arose from target location and idiosyncratic sources. The errors were largely unaffected by extreme variation in the arm's starting location. These data lend support to models of sensorimotor learning that focus on associations between postures and hand locations, but do not confirm a leading example of the class, exemplified by Kuperstein's (1988) circular reaction model. PMID- 15757846 TI - GH/IGF-I and bone resorption in vivo and in vitro. AB - IGF-I may act as one of several coupling agents by activating bone formation and bone resorption. In vivo studies in normal subjects, postmenopausal women and patients with excess or diminished GH production (acromegaly and GHD) indicate that both GH and IGF-I activate osteoclasts, but that GH has a more pronounced effect, independently of IGF-I. In vitro, GH and IGF receptors have been demonstrated on osteoclasts and both GH and IGF-I may directly modify osteoclast function and activity. In addition to direct effects on osteoclasts, GH and IGF-I may affect bone resorption indirectly by stimulating release of paracrine mediators that regulate osteoclastic resorption (cytokines). Critical for the bone resorptive process is the balance between OPG and RANKL, which is regulated by many systemic factors. In vivo and in vitro, GH/IGF-I may modulate this balance but these studies are difficult to interpret, reflecting the complexity of this system. Increased OPG expression may possibly protect against GH/IGF-I induced bone resorption and potentially be important for the long-term beneficial effects of GH replacement. Further studies investigating the OPG/RANKL ratio and system in experimental and transgenic GH/IGF models may clarify these issues. PMID- 15757847 TI - New insights into leptin resistance by modifying cytokine receptor signal transduction. PMID- 15757849 TI - Effect of ultrasound-guided interstitial laser photocoagulation on benign solitary solid cold thyroid nodules - a randomised study. AB - AIM: To evaluate the efficacy of ultrasound (US)-guided interstitial laser photocoagulation (ILP) on thyroid function, nodule size and patient satisfaction in benign solitary solid cold thyroid nodules by comparing one ILP session with no treatment in a prospective randomised study. MATERIALS AND METHODS: Thirty euthyroid outpatients with a benign solitary solid and a scintigraphically cold thyroid nodule causing local discomfort were assigned to one session of ILP (n = 15) or observation (n = 15) and followed for 6 months. Thyroid nodule volume and total thyroid volume were assessed by US and thyroid function was determined by routine assays before and during follow-up. Pressure and cosmetic complaints before and at 6 months were evaluated on a visual analogue scale. ILP was performed under US guidance and with an output power of 2.5-3.5 W. RESULTS: In the ILP group, the nodule volume decreased from 8.2 ml (6.1; 11.9) (median; quartiles) to 4.8 ml (3.0; 6.6) after 6 months (P = 0.001). The overall median reduction was 44% (37; 52), which correlated with a significant decrease in pressure symptoms as well as cosmetic complaints. In the control group, a non significant increase in median nodule volume of 7% (0; 34) after 6 months was seen. No major side-effects were seen in the ILP group. There was no correlation between thermal energy deposition and nodule volume reduction. Thyroid function was unaltered throughout. CONCLUSION: US-guided ILP, given as a single treatment, resulted in a satisfactory clinical response in the majority of patients with a benign solitary solid cold thyroid nodule, and may become a clinically relevant alternative to surgery in selected patients. PMID- 15757848 TI - Serum markers of GH and insulin action in 12-year-old children born small for gestational age. AB - OBJECTIVES: Our aim was to determine whether markers of growth hormone and insulin action differ between children born small for gestational age (SGA) and those born of an appropriate size for gestational age (AGA). DESIGN: Fifty-five SGA children and their 55 age- and sex-matched AGA control subjects were studied in a case-control setting at 12 years of age. METHODS: We examined serum concentrations of insulin-like growth factor (IGF)-I, IGF-II, IGF-binding protein (IGFBP)-1 and IGFBP-3, sex hormone binding globulin (SHBG), leptin, fasting insulin, and blood glucose. Insulin sensitivity was evaluated by the homeostasis model assessment for insulin resistance (HOMA-IR). RESULTS: The body mass index (BMI), sex, and puberty-adjusted mean serum IGF-I concentration was higher in the SGA than in the AGA children (303.4 vs 282.3 microg/l, P = 0.006). The mean serum concentrations of IGF-II, IGFBP-I, IGFBP-3, SHBG, fasting insulin, blood glucose and HOMA-IR did not differ between the SGA and the AGA group. The BMI, sex, and puberty-adjusted mean serum leptin concentration was lower in the SGA than in the AGA children (7.9 vs 10.1 microg/l, P = 0.037). In multiple logistic regression analysis, high HOMA-IR predicted high serum IGF-I levels in the SGA children (odds ratio 8.3; 95% confidence interval 1.7-41; P = 0.010), whereas in the AGA group HOMA-IR did not associate with the serum IGF-I level. CONCLUSIONS: The BMI, sex, and puberty-adjusted mean serum IGF-I concentration was significantly higher and the leptin concentration was lower in the SGA than in the AGA children. No differences were found in the indices of insulin action or sensitivity between the SGA and AGA children at 12 years of age. However, HOMA-IR strongly associated with serum IGF-I levels in the SGA children. PMID- 15757850 TI - Increased levels of serum osteoprotegerin in hypothyroid patients and its normalization with restoration of normal thyroid function. AB - Hypothyroidism is associated with increased morbidity from cardiovascular disease, and an increase in serum osteoprotegerin (OPG) has recently been reported to be associated with the severity of coronary heart disease and cardiovascular mortality. The present study was designed to examine whether hypothyroidism causes an increase in serum OPG, and to determine whether levothyroxine (L-T4) replacement therapy might suppress serum OPG levels in hypothyroid patients. Fifty-three hypothyroid patients with chronic thyroiditis and age- and sex-matched normal control subjects were examined for the levels of serum OPG and plasma von Willebrand factor (vWF), a vascular injury marker. Thirty-seven of the hypothyroid patients were further monitored for changes in these markers during 1 year in a euthyroid state induced by L-T4 replacement therapy. Baseline OPG was significantly higher in hypothyroid patients than in normal controls (4.51 +/- 0.50 vs 3.72 +/- 0.23 pmol/l (mean +/- S.E.); P = 0.0182). In multivariate analysis, baseline OPG was significantly associated with baseline levels of TSH (r = 0.280, P = 0.0162) and vWF (r = 0.626, P < 0.0001). During one year of L-T4 replacement therapy, hypothyroid patients showed a significant decrease in OPG levels from 4.35 +/- 0.51 to 3.48 +/- 0.26 pmol/l (P = 0.0166), a level comparable to normal controls. The change in serum OPG levels during L-T4 replacement therapy was significantly and independently associated in a negative fashion with baseline vWF (r = -0.503, P = 0.0014). This study suggested that the severity of hypothyroidism and vascular injury might have important independent roles in increasing the serum OPG level in hypothyroid patients. Furthermore, it was demonstrated that a sustained euthyroid state might have the potential to decrease the serum OPG level in hypothyroid patients and that the degree of vascular injury in the hypothyroid state is independently associated with a decrease in serum OPG during a 1-year normalization of thyroid function. PMID- 15757851 TI - Effects of levothyroxine treatment on biochemical and hemostasis parameters in patients with hypothyroidism. AB - OBJECTIVE: The aims of the study were to evaluate the disturbances in the coagulation system in patients with overt hypothyroidism (OH), to assess the effects of levothyroxine (LT4) on the coagulation parameters, and to determine whether subclinical hypothyroidism (SH) affects concentrations of coagulation markers and several biochemical parameters, thereby supporting early substitution. DESIGN: The study included 15 patients with SH (TSH levels 5-10 mU/l), 15 patients with OH and 15 euthyroid controls. METHODS: Blood urea nitrogen, creatinine, creatine phosphokinase, aspartate aminotransferase, lactate dehydrogenase, total-cholesterol, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol and triglyceride levels, and bleeding time, prothrombin time (PT), activated partial thromboplastin time (APTT), factor VIII activity, von Willebrand factor activity (vWF), platelet count and clotting time were evaluated just before and three months after the maintenance of euthyroidism with LT4 treatment. RESULTS: Factor VIII and vWF activities were lower in patients with SH than in controls (P < 0.01). Increased bleeding time, PT, APTT and clotting time and decreased factor VIII activity and vWF activity were observed in patients with OH when compared with controls. Bleeding time, PT, APTT and clotting time decreased and factor VIII activity, vWF and platelet count increased after LT4 in patients with OH. Increases in factor VIII activity and vWF (P < 0.01) were detected also in the SH group with treatment. CONCLUSIONS: OH is associated with significant abnormalities in clotting parameters which are reversed by LT4. In contrast, SH is associated with minor changes in factor VIII activity and vWF which are reversible by LT4. Serum lipids and other measured parameters are not improved by LT4 in patients with TSH < 10 mU/l and these data fail to demonstrate a need to treat such patients. PMID- 15757852 TI - FSH secretion predominates in vivo and in vitro in patients with non-functioning pituitary adenomas. AB - OBJECTIVE: Non-functioning pituitary adenomas (NFPAs) are characterised by the lack of symptoms of hormone hypersecretory syndromes but in vitro studies have demonstrated that tumour cells may stain for gonadotrophins and/or their alpha- or beta-subunits. In this study, we aimed to examine the pattern of secretion of LH and FSH from a series of pituitary adenomas cultured in vitro and where data were available to relate the results to pre-operative serum gonadotrophin levels. METHODS: The in vitro secretion of LH and FSH was measured from 46 cultured NFPAs and compared with pre-operative serum gonadotrophin levels in 38 patients. Peritumorous 'normal' pituitary cell cultures from 20 additional pituitary tumour patients were used for comparison with the NFPA group. RESULTS: A median pre operative LH:FSH ratio of 0.33:1 was found in 38 patients with NFPAs. Preferential secretion of FSH was also documented from media of 46 NFPAs cultured in vitro with a median LH:FSH ratio of 0.32:1. A significant correlation (r = 0.43, P < 0.01) was observed between serum and media levels of FSH but not LH. Peritumorous 'normal' pituitary cells released LH and FSH in a reversed ratio (median LH:FSH ratio = 3.6:1, P < 0.01 compared with NFPAs). CONCLUSIONS: This study has evaluated pre-operative serum gonadotrophin levels and in vitro release of hormones in cultures of surgically removed tissue from patients with NFPAs. The data suggest preferential secretion of FSH occurs both in vitro and in vivo. By demonstrating that NFPAs cultured in vitro reflect the in vivo situation of preferential secretion of FSH, it may be possible in future to perform functional studies using this system to elucidate the cellular and molecular mechanisms involved in the development of an imbalance in gonadotroph cells preferentially overproducing FSH in NFPAs. PMID- 15757853 TI - The natural history of post-traumatic neurohypophysial dysfunction. AB - BACKGROUND AND OBJECTIVES: Posterior pituitary function remains poorly investigated after traumatic brain injury (TBI). We report the results of a study designed to prospectively define the natural history of post-traumatic diabetes insipidus (DI) and syndrome of inappropriate antidiuretic hormone secretion (SIADH) using standard reliable methodology. DESIGN AND METHODS: 50 consecutive patients with severe or moderate TBI (initial Glasgow Coma Scale (GCS) score 3/15 13/15) were prospectively studied on three occasions: at the acute phase and at 6 months and at 12 months following TBI. In the acute phase, DI was diagnosed either by the presence of hypernatraemia in association with hypotonic polyuria or by the water-deprivation test (WDT) and, at 6 and 12 months by the WDT in all patients. Normative data on response to the WDT were obtained from healthy matched volunteers. Functional outcome was assessed using the Glasgow Outcome Scale (GOS). RESULTS: 13 patients (26%) had DI in the acute post-TBI phase, of whom nine patients recovered by 6 months and one additional patient recovered by 12 months. Of the remaining three patients with permanent DI, two had partial vasopressin deficiency. Acute-phase peak plasma osmolality correlated negatively with the initial GCS scores (r = -0.39, P = 0.005) and with the GOS scores (r = 0.45, P = 0.001). Seven patients had SIADH in the acute phase of TBI but none did at 6 or 12 months. No new cases of DI or SIADH were noted after the acute phase. CONCLUSION: This prospective study shows that posterior pituitary dysfunction is common following TBI. Most cases recover completely but there is an appreciable frequency of long-term DI which can be subtle and should be recognized and managed appropriately. PMID- 15757854 TI - The outcome of surgery in 668 patients with acromegaly using current criteria of biochemical 'cure'. AB - BACKGROUND AND AIM: The aim of this study was to illustrate the present role of transsphenoidal surgery as primary therapy in GH-secreting adenomas, and to compare the results concerning control of disease with previous series using older criteria of cure. METHOD: We report on a consecutive series of 688 acromegalic patients treated over a time period of 19 years. Biochemical cure was defined as normalisation of basal GH level, suppression of GH levels to below 1 ng/ml during an oral glucose load and normalisation of IGF-I levels. Of the 506 patients undergoing primary transsphenoidal surgery, a total of 57.3% postoperatively fulfilled the criteria used. RESULTS: The rate of biochemical 'cure' correlated with the magnitude of the initial GH levels, the tumour size and invasion. The overall complication rate was below 2%. Mortality in this series was 0.1% (1 of 688). During a follow-up period of 10.7 years only two recurrences (0.4%) occurred. However, in the patients treated by transcranial surgery and by repeat surgery the cure rate was found to be relatively low (5.2 and 21.3% respectively). CONCLUSIONS: These data suggest that surgery remains with very few exceptions the primary treatment of acromegaly for (i) a high cure rate, (ii) low morbidity, (iii) low recurrence rate and (iv) immediate decline of GH. Based on current criteria of cure, recurrences are uncommon. However, cure by surgery alone is improbable in patients harbouring extended, invasive tumours with high secretory activity, in whom further adjuvant treatment is mandatory. PMID- 15757855 TI - Evidence for altered adipocyte function in polycystic ovary syndrome. AB - BACKGROUND: Adipocytokines are produced by adipose tissue and have been thought to be related to insulin resistance and other health consequences. We measured leptin, adiponectin, and resistin simultaneously in women with polycystic ovary syndrome (PCOS) and age- and weight-matched controls. Our hypothesis was that these simultaneous measurements would help determine whether adipocytokine secretion is abnormal in PCOS independent of body mass and whether these levels are related to insulin resistance as well as other hormonal changes. METHODS: Fifty-two women with PCOS and 45 normal ovulatory women who were age- and weight matched were studied. Blood was obtained for adipocytokines (leptin, adiponectin, and resistin) as well as hormonal parameters and markers of insulin resistance as assessed by the quantitative insulin-sensitivity check index. Body mass index (BMI) was stratified into obese, overweight, and normal subgroups for comparisons between PCOS and controls. RESULTS: Adiponectin was lower (P < 0.05) and resistin was higher (P < 0.05) while leptin was similar to matched controls. Breakdown of the groups into subgroups showed a strong body mass relationship for leptin with no changes in resistin although adiponectin was lower in PCOS, even controlling for BMI. In controls, leptin and adiponectin and leptin and resistin correlated (P < 0.05) but not in PCOS. In controls, all adipocytokines correlated with markers of insulin resistance but not in PCOS. CONCLUSIONS: When matched for BMI status, decreased adiponectin in PCOS represent the most marked change. This alteration may be the result of altered adipose tissue distribution and function in PCOS but no correlation with insulin resistance was found. PMID- 15757856 TI - Relationship between inhibin A and B, estradiol and follicle growth dynamics during ovarian stimulation in normo-ovulatory women. AB - OBJECTIVE: To investigate the relationship between serum concentrations of inhibin A, inhibin B and estradiol (E(2)) and the number of developing follicles during the administration of exogenous follicle-stimulating hormone (FSH) in various regimens in normo-ovulatory volunteers and to evaluate if inhibins act as suitable markers for the number of developing follicles during ovarian stimulation. DESIGN AND METHODS: Serial hormone determinations and assessment of follicle numbers were carried out during unstimulated cycles and during various interventions with exogenous FSH. Subjects were randomized for FSH administration into the following groups: a single high dose (375 IU) during the early follicular phase (group A), 5 consecutive low doses (75 IU/day) starting in the mid follicular phase (group B) or daily low doses (75 IU/day) during the early to late follicular phase (starting on cycle days 3, 5 or 7; groups C, D and E respectively). RESULTS: Extending the FSH window increases the number of small antral follicles and hence inhibin B serum concentrations. If such an intervention results in multi-follicular growth, mid follicular phase inhibin B (P = 0.001) as well as late follicular phase inhibin B and inhibin A levels are significantly (P < 0.05 and P < 0.01 respectively) increased compared with mono follicular cycles or the natural cycle. Although mid follicular inhibin B levels correlated well with the number of small antral (P < 0.05) and pre-ovulatory (P < 0.001) follicles in the late follicular phase, mid follicular inhibin A and estradiol serum concentrations only correlated with the number of pre-ovulatory follicles (P < 0.001 and P < 0.01 respectively). CONCLUSIONS: The present data extend our understanding of the relationship between follicle dynamics, serum inhibins and FSH during ovarian hyperstimulation. However, although mid follicular inhibin B does correlate with the number of developing follicles, it does not facilitate the identification of women at risk for multiple follicle development. PMID- 15757857 TI - Changes in serum inhibin B during normal male puberty. AB - OBJECTIVE: In adult men, inhibin B (InhB) regulates FSH secretion by a negative feedback. The aims of this study were to evaluate the changes of InhB during puberty in the male and the relationship between InhB and FSH, LH, testosterone and testicular volume. DESIGN: Cross-sectional study. METHODS: InhB was measured using a two-site ELISA in 100 healthy boys subdivided by their pubertal development according to Tanner into five groups of 20. RESULTS: During puberty we observed an increase of InhB level (G1 = 84.3 pg/ml, G3 = 132.2 pg/ml, G5 = 206.1 pg/ml). In G1, InhB correlated positively with FSH (P = 0.0001), LH (P = 0.005), testosterone (P = 0.001) and testicular volume (P = 0.007); in G5, InhB correlated inversely with FSH (P = 0.001) and LH (P = 0.045) and directly with testicular volume (P = 0.013). The multivariate analysis demonstrated that: in G1, FSH is the most important, and testosterone the second most significant, stimulus for InhB increase; in G2 only FSH has a positive effect on InhB variation; in G3 only mean testicular volume fits the model (G1-G3: InhB dependent variable); considering the FSH dependent variable, in G4, InhB is the most important stimulus for FSH decrease and mean testicular volume is a secondary directly proportional variable; in G5, only InhB shows a significant inverse relationship with FSH. CONCLUSIONS: During puberty there is a regular increase of InhB. In the first phases of gonadal maturation, InhB and FSH correlate positively, while in mid-late stages the relationship is inverse. We found that in mid-puberty (G3-G4), the serum concentration of InhB increases, as its inverse relationship with FSH is being established and hence spermatogenesis. PMID- 15757858 TI - Late follicular phase administration of mifepristone suppresses circulating leptin and FSH - mechanism(s) of action in emergency contraception? AB - OBJECTIVE: Low dose mifepristone (RU486) is highly effective in emergency post coital contraception (EC), although the mechanism(s) of action remains unclear. We studied the endocrine actions of 10 mg mifepristone administered orally as a single dose to eight healthy volunteers (aged 20-45 years) during the late follicular phase. METHODS: Serum levels of LH, FSH, oestradiol, progesterone, leptin, mifepristone, cortisol, and gluco-corticoid bioactivity (GBA) were measured before and 1, 2, 4 and 8 h after ingestion of mifepristone on cycle day 10 or 11 (study day 1), and follow-up was continued for 10 days. Ovarian ultrasonography was performed on study days 1 and 7. Similar measurements were carried out during a control cycle. RESULTS: Mifepristone postponed ovulation, as evidenced by a 3.4+/-1.1 day (means+/-s.d.) delay (P < 0.005) in the LH surge and 3.6+/-4.0 day prolongation of the treatment cycle (P = 0.08). During the mifepristone cycle, an LH surge was displayed by five subjects when serum mifepristone levels had declined to 9.5+/-7.1 nmol/l. During the day of mifepristone administration, circulating GBA (P < 0.001) and leptin (P < 0.001) levels declined. On the day after mifepristone administration, mean serum FSH and leptin levels were lower than pretreatment values (3.8+/-1.8 IU/l vs 5.2+/-1.1 IU/l, n = 7, P < 0.05; 28.9+/-6.7 microg/l vs 33.2+/-9.0 microg/l, n = 7, P < 0.05 respectively), and the corresponding difference in the mean serum oestradiol concentration was borderline (452+/-252 pmol/l vs 647+/-406 pmol/l, n = 7, P = 0.056). In contrast to the control cycle, individual leptin levels declined during the follow-up after ingestion of mifepristone (n = 8, P < 0.01). CONCLUSIONS: These data showed that the commonly employed dose of mifepristone for EC delays ovulation and prolongs the menstrual cycle, when given during the late follicular phase. The mechanism of action of mifepristone may include a reduction of FSH secretion via a decrease in circulating leptin. PMID- 15757859 TI - Androgen receptor gene CAG and GGC repeat lengths in cryptorchidism. AB - OBJECTIVE: Cryptorchidism is the most common congenital birth defect in male children, and accumulating evidence suggests that genetic abnormalities may be associated with it. The androgen receptor has two polymorphic sites in exon 1, with different numbers of CAG and GGC repeats, resulting in variable lengths of polyglutamine and polyglycine stretches. Longer CAG repeats result in a reduced androgen receptor transcriptional activity, but the role of the GGC triplets is less clear. In this study we analysed CAG and GGC repeat lengths in men with a history of cryptorchidism, associated or not with impairment of sperm production, in comparison with normal fertile subjects. METHODS: We analysed CAG and GGC repeat lengths in a group of 105 ex-cryptorchid men in comparison with 115 fertile non-cryptorchid men. RESULTS: No difference was found between patients and controls in the mean and median values, and in distribution of CAG and GGC, when considered separately. However, the analysis of the joint distribution of CAG and GGC showed that some combinations are significantly more frequent in men with bilateral cryptorchidism (who frequently presented severe testiculopathies), in a manner similar to that found in idiopathic infertile subjects. CONCLUSIONS: Although further studies are needed to elucidate the possible role of specific CAG/GGC combinations as a causative factor, these data suggest a possible association between androgen receptor gene polymorphisms and cryptorchidism. PMID- 15757861 TI - Diabetes and premature menopause: is their co-existence detrimental to the skeleton? AB - OBJECTIVE: Premature menopause is a known risk factor for osteoporosis, whilst the influence of type 2 diabetes on bone mineral density (BMD) is still controversial. DESIGN AND METHODS: BMD values assessed by dual-energy X-ray absorptiometry (DXA) in L2-L4 vertebrae and the femoral neck (FN) of 40 diabetic women with premature menopause (D-EMP) were compared with those of 60 non diabetic, prematurely menopausal women (EMP) and 60 diabetic women with normal menopause (D-NMP) who had been matched by age and body mass index (BMI). In all women, the time elapsed since menopause ranged between 10 and 25 years and the duration of diabetes exceeded 75% of the postmenopausal time period. The age of D EMP women was 58.7+/-5 years (mean+/-1 s.d.), age at menopause 39.5+/-2.7, years since menopause 18.6+/-4.9, BMI 27.8+/-4.3 kg/m(2) and duration of diabetes 13.9+/-3.9 years. RESULTS: Vertebral BMD values of D-EMP women were significantly higher than those of EMP women (0.908+/-0.135 vs. 0.817+/-0.14 g/cm(2), P = 0.002), although there was no significant difference between D-EMP and D-NMP women (0.886+/-0.15 g/cm(2)). No significant differences were observed in FN BMD values between all groups. Age-adjusted BMD values (Z scores) of D-EMP women were higher than EMP women in both anatomic sites (P < 0.01), but did not differ from D-NMP women. In contrast to the other two groups, no statistically significant correlation was observed in D-EMP women between the BMD values of either anatomic area and the time elapsed since menopause. HbA(1c) values were positively correlated only to vertebral BMD values of the D-EMP group (P < 0.05). No correlation was observed between the BMD values and the duration of diabetes either in D-EMP or in D-NMP women. CONCLUSIONS: Type 2 diabetes seems to positively affect the mineral density of the trabecular bone in women with premature menopause. The duration of diabetes does not appear to influence bone mass. PMID- 15757860 TI - The effects of exercise and adipose tissue lipolysis on plasma adiponectin concentration and adiponectin receptor expression in human skeletal muscle. AB - OBJECTIVE: It has been suggested that adiponectin regulates plasma free fatty acid (FFA) clearance by stimulating FFA uptake and/or oxidation in muscle. We aimed to determine changes in plasma adiponectin concentration and adiponectin receptor 1 and 2 mRNA expression in skeletal muscle during and after prolonged exercise under normal, fasting conditions (high FFA trial; HFA) and following pharmacological inhibition of adipose tissue lipolysis (low FFA trial; LFA). Furthermore, we aimed to detect and locate adiponectin in skeletal muscle tissue. METHODS: Ten subjects performed two exercise trials (120 min at 50% VO(2max)). Indirect calorimetry was used to determine total fat oxidation rate. Plasma samples were collected at rest, during exercise and during post-exercise recovery to determine adiponectin, FFA and glycerol concentrations. Muscle biopsies were taken to determine adiponectin protein and adiponectin receptor 1 and 2 mRNA expression and to localise intramyocellular adiponectin. RESULTS: Basal plasma adiponectin concentrations averaged 6.57+/-0.7 and 6.63+/-0.8 mg/l in the HFA and LFA trials respectively, and did not change significantly during or after exercise. In the LFA trial, plasma FFA concentrations and total fat oxidation rates were substantially reduced. However, plasma adiponectin and muscle adiponectin receptor 1 and 2 mRNA expression did not differ between trials. Immunohistochemical staining of muscle cross-sections showed the presence of adiponectin in the sarcolemma of individual muscle fibres and within the interfibrillar arterioles. CONCLUSION: Plasma adiponectin concentrations and adiponectin receptor 1 and 2 mRNA expression in muscle are not acutely regulated by changes in adipose tissue lipolysis and/or plasma FFA concentrations. Adiponectin is abundantly expressed in muscle, and, for the first time, it has been shown to be present in/on the sarcolemma of individual muscle fibres. PMID- 15757862 TI - Plasma chromogranin A in patients with autoimmune chronic atrophic gastritis, enterochromaffin-like cell lesions and gastric carcinoids. AB - OBJECTIVE: In atrophic body gastritis (ABG) chronic hypergastrinaemia stimulates enterochromaffin-like (ECL) cell proliferation with development of cell hyperplasia, dysplasia and possibly type-1 gastric carcinoids. As circulating chromogranin A (CgA) levels are a marker of neuroendocrine tumours, we evaluated the clinical usefulness of CgA assay in ABG patients to detect those with carcinoids. DESIGN AND METHODS: Plasma CgA levels were measured using a commercial ELISA in 45 healthy volunteers, nine patients with type-1 gastric carcinoids and 43 consecutive ABG patients (21 without and 22 with ECL cell hyperplasia/dysplasia). RESULTS: CgA levels were significantly higher in ABG patients with and without gastric carcinoids than in healthy subjects (P < 0.001). The highest values occurred in patients with carcinoids (median (interquartile range): 58.1 (44.5-65.3) U/l) and with ECL cell hyperplasia/dysplasia (35.5 (31.8-48.65) U/l) but there were no significant differences in CgA among the various subgroups of ABG patients classified according to ECL cell status. Nevertheless, in ABG patients without carcinoids CgA values correlated with the presence and severity of ECL cell lesions (r(s) = 0.428, P < 0.01). The sensitivity and specificity of the CgA assay in identifying patients with carcinoids were 100 and 23% respectively. CONCLUSIONS: CgA plasma levels reflect the histological degree of ECL cell lesions in patients with ABG but the assay specificity is too low to detect among these patients those with gastric carcinoids. PMID- 15757863 TI - Aging affects the retinoic acid and the triiodothyronine nuclear receptor mRNA expression in human peripheral blood mononuclear cells. AB - BACKGROUND: Inadequate retinoid status has often been described as occurring with aging. Moreover, subclinical hypothyroid status has also been evoked in the elderly. Several studies performed in animals have described the crucial incidence of age-related hypo-functioning of retinoid and thyroid signalling pathways, particularly in the brain. OBJECTIVE: The aim of the present study was to clarify whether aging modifies retinoid and thyroid signalling in humans. METHODS: Using real-time RT-PCR the relative amount of mRNA of the retinoid (RARalpha, RARgamma and RXRalpha) and thyroid (TRalpha and TRbeta) nuclear receptors in peripheral blood mononuclear cells (PBMC) of young (24-57 years old, n = 22) compared with elderly (69-90 years old, n = 24) healthy subjects was quantitated. Classical plasma parameters used to characterize the retinoid and thyroid status - retinol (ROH), retinol-binding protein (RBP), free triiodothyronine (FT3) and thyroxine (FT4), thyroid-stimulating hormone (TSH) and transthyretin (TTR) - were also assessed. RESULTS: RARgamma expression was significantly decreased in elderly versus young subjects while no modification of the retinoid-related plasma parameters ROH and RBP were emphasized by aging. Concerning thyroid criteria, the elderly exhibited an increase in TSH concentration (+39%) without significant modifications of FT3 and FT4, which indicated an age-related sub-clinical hypothyroidism. Concurrently, the amount of TR mRNA (alpha as well as beta subtypes) was significantly decreased in the elderly. CONCLUSION: These data constitute the first evidence of an age-related hypo-activation of the retinoid and thyroid nuclear pathways in PBMC. Further study of the possible association between the expression of the retinoid and thyroid nuclear receptors and age-related cognitive alterations in humans would be interesting. PMID- 15757864 TI - Distinction in gene expression profiles demonstrated in parathyroid adenomas by high-density oligoarray technology. AB - OBJECTIVES: Somatic deletion of chromosome 11q13 is the most frequent genetic aberration in parathyroid adenoma. To gain further insight into the genetic etiology of parathyroid tumor development, we examined a comprehensive gene expression profile of parathyroid adenomas and normal parathyroid tissues. The results were then evaluated with respect to differences between adenomas and normal parathyroid tissue, and to the presence of loss of heterozygosity (LOH) in chromosomal region 11q13. DESIGN AND METHODS: Sporadic parathyroid adenomas and normal parathyroids were hybridized against HG-U95Av2 oligonucleotide arrays (Affymetrix) containing a total of 12,625 probe sets. Quantitative real-time PCR (QRT-PCR) was performed in a larger series of parathyroid adenomas, in order to con-firm the microarray results. RESULTS: Cyclin D1 and c-Jun showed increased expression in adenomas vs normal parathyroids by microarray analysis and QRT-PCR, suggesting an oncogenic role of these genes in parathyroid tumor development. At unsupervised hierarchical clustering, the adenomas fell into two groups: Group I adenomas were characterized by 11q13 LOH, while Group II adenomas lacked this abnormality. In addition, a t-test analysis identified largely non-overlapping genes with differential expression in the tumors subgroups; e.g. in Group I tumors the putative oncogene ENC 1 was found highly over-expressed vs Group II adenomas. CONCLUSIONS: The microarray analyses revealed partly distinctive and partly common expression profiles in parathyroid adenomas with and without 11q13 LOH. In addition, approximately half of the under-expressed genes were mapped to chromosome 11, in agreement with a dose effect following loss of this chromosome. PMID- 15757865 TI - Empirical estimation of free testosterone from testosterone and sex hormone binding globulin immunoassays. AB - BACKGROUND: The growing interest in measuring blood free testosterone (FT) is constrained by the unsuitability of the laborious reference methods for wider adoption in routine diagnostic laboratories. Various alternative derived testosterone measures have been proposed to estimate FT from either additional assay steps or calculations using total testosterone (TT) and sex hormone-binding globulin (SHBG) measured in the same sample. However, none have been critically validated in large numbers of blood samples. METHODS: We analyzed a large dataset comprising over 4000 consecutive blood samples in which FT as well as TT and SHBG were measured. Dividing the dataset into samples with blood TT above and below 5 nM, using a bootstrap regression modeling approach guided by Akaike Information Criterion for model selection to balance parsimony against reduction of residual error, empirical equations were developed for FT in terms of TT and SHBG. RESULTS: Comparison between the empirical FT equations with the laboratory FT measurements as well as three widely used calculated FT methods showed the empirical FT formulae had superior fidelity with laboratory measurements while previous FT formulae overestimated and deviated systematically from the laboratory FT values. CONCLUSION: We conclude that these simple, assumption-free empirical FT equations can estimate accurately blood FT from TT and SHBG measured in the same samples with the present assay methods and have suitable properties for wider application to evaluate the clinical utility of blood FT measurements. PMID- 15757866 TI - Acute actions of testosterone on contractile function of isolated rat ventricular myocytes. AB - Variation between the sexes in cardiac function have been established. The extent to which sex hormones are responsible for these differences is unclear. The current study was designed to determine whether testosterone acts acutely to enhance contractility of cultured rat ventricular myocytes. Following a 24-h treatment with testosterone (1 microM), isolated rat ventricular myocytes display a 21% increase (P < 0.01) in peak shortening and an 18% decrease (P < 0.02) in time to peak shortening. In accordance with this change, testosterone treatment produced an 18% decline (P < 0.002) in the time to relengthening when compared to vehicle-treated controls. These results provide the first evidence that short term androgen exposure acts directly to stimulate contractility of isolated rat ventricular myocytes and thus may play a role in regulating cardiac performance in males and thereby contribute to sex differences in cardiac function. PMID- 15757867 TI - Suppressor of cytokine signaling gene expression in human pancreatic islets: modulation by cytokines. AB - OBJECTIVE: Suppressor of cytokine signaling (SOCS) proteins negatively regulate signal transduction of several cytokines. Since cytokines participate in the pancreatic islet damage in type 1 diabetes, the aim of our study was to investigate the expression of SOCS-1, -2 and -3 in isolated human islets, in basal conditions and after exposure, in vitro, to a combination of interferon (IFN)-gamma, interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha cytokines and in control and in type 1 diabetic human pancreata, to establish (i) whether SOCS molecules are constitutively expressed in human pancreatic islets and (ii) whether their expression can be modulated in vitro by proinflammatory cytokines or ex vivo by an islet inflammatory process. METHODS: Gene expression of SOCS-1, -2 and -3 was evaluated by RT-PCR in untreated and cytokine-treated isolated human pancreatic islets and their protein expression by immunohistochemistry in control and in type 1 diabetic human pancreata paraffin embedded sections. RESULTS: We found that SOCS-1, -2 and -3 mRNA is constitutively, although weakly, expressed in human pancreatic islets, similar to the expression observed in control pancreata by immunohistochemistry. SOCS-1, -2 and -3 mRNA expression was strongly increased in human islets after exposure, in vitro, to IFN-gamma, IL-1beta and TNF-alpha. Accordingly, an intense and islet specific immunohistochemical staining for all three SOCS was detected in pancreata from type 1 diabetic patients. CONCLUSION: SOCS-1, -2 and -3 genes are constitutively expressed in human pancreatic islets; their expression increases after exposure to proinflammatory cytokines and during an autoimmune inflammatory process, raising the possibility that these molecules act as key regulators of cytokine signaling in pancreatic islets. PMID- 15757869 TI - Factors affecting the likelihood that people with intellectual disabilities will gain employment. AB - The study aimed to identify factors that may affect the likelihood that people with intellectual disabilities will find employment through a supported employment agency. A retrospective analysis of the files of the last 200 individuals to use such an agency was carried out. Variables were identified that may influence outcome, such as demographic factors, job preferences and identified support needs. A specific rating of staff assessed client motivation was developed from written comments made by staff. Staff assessed motivation after two weeks on the scheme was found to be the only significant predictor of employment outcome. Motivation levels decreased for all groups of participants while at the agency, irrespective of work outcome. This study highlights that a greater emphasis on motivation within employment agencies may increase the chances of people with intellectual disability finding work. PMID- 15757871 TI - Sibshops: an evaluation of an interagency model. AB - The study evaluates the effectiveness of a sibling support programme (Sibshops) involving three disability agencies in Cork, Ireland. Qualitative and quantitative data were obtained using semi-structured interviews and the Piers Harris Children's Self-Concept Scale with siblings, together with parent feedback. Results from the Piers-Harris showed that there was no significant increase in sibling self-esteem following attendance at the Sibshops. However, the interviews revealed that the majority of siblings enjoyed and benefited from the Sibshops. Parents reported satisfaction with the Sibshops and felt that their children had benefited. Reflections on the experience of working on an interagency basis are outlined. Recommendations are made regarding further research and development in the organization of Sibshops and in staffing and staff training. PMID- 15757870 TI - Patterns of leisure participation among adolescents with a mild intellectual disability. AB - The aim of this study was to investigate the leisure and recreational activities of a cohort of 34 students attending a full-time special school as reported by the students and their parents. Leisure activities which were mostly solitary and passive in nature were identified as those being most commonly engaged in. Barriers to leisure were also identified, with 'access to' and 'location of' the leisure facilities being barriers perceived by both students and parents. Data on aspects of leisure use, e.g. enjoyment, participation, assistance required, socialization and choice, were also collected. PMID- 15757872 TI - Self-reported smoking and alcohol use among adolescents with intellectual disabilities. AB - Self-reported smoking and alcohol use were examined among 95 adolescents (aged 11 15) with intellectual disabilities and 4069 adolescents who did not have intellectual disabilities. Results indicated that adolescents with intellectual disabilities reported increased rates of smoking and decreased rates of using alcohol at least once a month. There were no marked differences between the two groups on smoking seven or more cigarettes a day, having ever smoked at all and having ever used alcohol. Elevated levels of smoking appeared to be an artefact of increased rates of poverty among adolescents with intellectual disabilities. Distinct patterns of risk were associated with smoking and alcohol use among adolescents with intellectual disabilities. Smoking was closely associated with the experience of poverty and adolescent mental health. Alcohol use was associated with less punitive child management practices and carer mental health. Results are discussed in the context of health promotion. PMID- 15757873 TI - Suspects who have a learning disability: police perceptions toward the client group and their knowledge about learning disabilities. AB - More than a million people in the UK have a learning disability. A small but significant proportion of these people will come into contact with the criminal justice system because they have offended or have been accused of an offence. This article reports on the perceptions of police officers toward this potentially vulnerable group and their knowledge of issues related to learning disabilities. Interview data were collected from eight police sergeants employed by Humberside Police and analysed using qualitative techniques. The article concludes that further training in the subject area is required. This is likely to be successful in the force under study due to their promotion of positive perceptions toward people with learning disabilities. PMID- 15757874 TI - Project advances health system's clinical pharmacy services. PMID- 15757875 TI - Report urges scientific standards for alternative medicine. PMID- 15757876 TI - Conference focuses on health care reform, lack of technology. PMID- 15757877 TI - Dietary supplements for female sexual dysfunction. PMID- 15757878 TI - Collaborating for successful medication therapy management programs. PMID- 15757879 TI - Summary of the Executive Sessions on Medication Therapy Management Programs: Bethesda, Maryland, June 14 and August 18, 2004. PMID- 15757880 TI - Current and emerging treatment options for venous thrombosis: a case discussion. PMID- 15757881 TI - Nesiritide for secondary pulmonary hypertension in patients with end-stage heart failure. AB - PURPOSE: The impact of adding nesiritide to standard therapy and positive inotropic agents in patients with end-stage heart failure and secondary pulmonary hypertension (PH) was studied. METHODS: Patients included in this retrospective study were 18 years of age or older, admitted to the hospital with PH secondary to end-stage heart failure (New York Heart Association functional class IV), had received a pulmonary artery catheter, had been treated with nesiritide because of inadequate hemodynamic response to previous therapy (pulmonary capillary wedge pressure [PCWP], >18 mm Hg), and had shown minimal symptomatic benefit from standard heart-failure therapy, continuous infusions of loop diuretics, and positive inotropic agents (milrinone or dobutamine or both). The primary endpoint was change in PCWP. Secondary endpoints included change in mean pulmonary artery pressure (MPAP), change in cardiac index (CI), change in mean arterial pressure (MAP), change in serum creatinine (SCr) concentration, and occurrence of symptomatic hypotension. RESULTS: The study included 33 patients. Mean PCWP was reduced by 31.1% with the addition of nesiritide to previous therapy (p < 0.0001). Significant improvements in other hemodynamic variables, including MPAP (15.6% reduction) and CI (13.0% increase), were also observed. MAP was reduced significantly (by 15.2%), but SCr concentration did not change. There were five episodes of symptomatic hypotension. All patients exhibited relief of dyspnea symptoms. CONCLUSION: The addition of nesiritide to standard therapy and positive inotropic agents improved hemodynamic measures and clinical symptoms in patients with end-stage heart failure and secondary pulmonary hypertension. PMID- 15757882 TI - Health care costs for schizophrenia patients started on olanzapine versus risperidone. AB - PURPOSE: The change in direct medical costs for schizophrenia patients who were started on olanzapine or risperidone and who were privately insured was studied. METHODS: A retrospective analysis of 1996-1999 data from the databases representing the health care experiences of individuals employed by large organizations and their dependents was performed. The sample included all individuals with a drug claim for olanzapine or risperidone, a claim with a schizophrenia diagnosis within 90 days of the drug claim, no claim for the same drug in the prior six months, and continuous health-plan enrollment for 12 months before and after the prescription. RESULTS: The sample included 162 patients initiated on olanzapine and 119 patients initiated on risperidone. Demographic and clinical profiles were not significantly different between groups. Annual schizophrenia-related prescription and outpatient costs increased following initiation on olanzapine or risperidone compared with the pre-initiation period. This was partially offset by a decrease in inpatient expenditures. Olanzapine initiators had higher outpatient drug expenditures than risperidone initiators in the 12 months following initiation (adjusted means, $2105 versus $1934) (p < 0.05), but there was no significant difference between groups in total schizophrenia-related payments ($5251 versus $4950). CONCLUSION: The total health care expenditure related to treating schizophrenia was similar between privately insured patients who were initiated on olanzapine and patients who were started on risperidone. PMID- 15757883 TI - Outpatient treatment of venous thromboembolic disease based in an emergency department. AB - PURPOSE: The efficacy, safety, and patient satisfaction of an emergency department (ED)-based outpatient program providing treatment for venous thromboembolic disease (VTD) were studied. METHODS: A prospective cohort study of patients enrolled in a hospital outpatient VTD treatment program was conducted from June 1999 to September 2003. Endpoints included recurrent VTD at three and six months following discharge from the program, minor and major bleeding, and thrombocytopenia during the acute phase of therapy. Patient satisfaction was assessed with an 18-question survey that was mailed to all patients following discharge from the program. RESULTS: A total of 162 patients were included. Of the 142 evaluable patients, no patient experienced recurrent VTD at three months, while at six months, 2 patients (1.4%; 95% confidence interval [CI], 0.2-5.0%) had recurrence. No patient experienced major bleeding or thrombocytopenia, while 4 (2.5%; 95% CI, 0.8-6.2%) had minor bleeding. Ninety-seven percent of the patients were comfortable having their condition treated on an outpatient basis, and 85.1% felt it was more convenient to return to the hospital daily for medications and assessment than to be admitted to the hospital. Overall, 97.7% of the respondents were satisfied or very satisfied with the treatment received in the outpatient program, and 94.0% said they would enroll again if future treatment was indicated. CONCLUSION: An ED-based outpatient VTD treatment program was safe and effective and appeared to achieve a high level of patient satisfaction. PMID- 15757884 TI - Evaluating the educational content of direct-to-consumer fulfillment materials. AB - PURPOSE: The educational content of direct-to-consumer (DTC) fulfillment materials was evaluated. METHODS: A list of prescription drug products advertised to consumers via broadcast media from August 1997 through April 20, 2002, was obtained from the Food and Drug Administration (FDA). The prescription products were categorized by disease state on the basis of their FDA-approved indications. Eight disease states were selected for analysis purposes and included acne, allergic rhinitis, depression, diabetes, hypercholesterolemia, migraine, overactive bladder, and osteoporosis. A total of 31 products were advertised directly to consumers for the eight targeted disease states, 26 of which met the inclusion criteria. The educational content of the advertisements for these 26 products was assessed by analyzing the materials' consistency, instructiveness, and consumer orientation. RESULTS: Two of the 26 materials analyzed contained claims that potentially broadened a drug's indication from that listed in the FDA approved labeling. The majority of materials listed the condition name (92%), symptom information (77%), the drug's mechanism of action (65%), the drug's time to onset of action (54%), and supportive behaviors (62%). Twenty of 24 DTC fulfillment materials (83%) were not written at the reading level of eighth grade or lower. Fifteen of the 26 mailings contained educational diagrams, 52% of which met the criteria for necessity, and a greater percentage met the criteria for suitability (90%), familiarity (86%), overall layout (88%), single concept (86%), and lack of distracting elements (100%). CONCLUSION: DTC fulfillment materials appear to have more educational content than DTC print advertisements but are still overwhelmingly deficient in meeting the recommended sixth to eighth-grade reading level. PMID- 15757885 TI - Impact of an interdisciplinary team on drug therapy outcomes in a geriatric clinic. PMID- 15757887 TI - Albumin for fluid resuscitation: implications of the Saline versus Albumin Fluid Evaluation. PMID- 15757886 TI - Stability of ibuprofen in injection solutions. PMID- 15757888 TI - Status of and pharmacists' role in patient-delivered partner therapy for sexually transmitted diseases. PMID- 15757889 TI - c-Myc suppresses p21WAF1/CIP1 expression during estrogen signaling and antiestrogen resistance in human breast cancer cells. AB - Estrogen rapidly induces expression of the proto-oncogene c-myc. c-Myc is required for estrogen-stimulated proliferation of breast cancer cells, and deregulated c-Myc expression has been implicated in antiestrogen resistance. In this report, we investigate the mechanism(s) by which c-Myc mediates estrogen stimulated proliferation and contributes to cell cycle progression in the presence of antiestrogen. The MCF-7 cell line is a model of estrogen-dependent, antiestrogen-sensitive human breast cancer. Using stable MCF-7 derivatives with inducible c-Myc expression, we demonstrated that in antiestrogen-treated cells, the elevated mRNA and protein levels of p21(WAF1/CIP1), a cell cycle inhibitor, decreased upon either c-Myc induction or estrogen treatment. Expression of p21 blocked c-Myc-mediated cell cycle progression in the presence of antiestrogen, suggesting that the decrease in p21 is necessary for this process. Using RNA interference to suppress c-Myc expression, we further established that c-Myc is required for estrogen-mediated decreases in p21(WAF1/CIP1). Finally, we observed that neither c-Myc nor p21(WAF1/CIP1) is regulated by estrogen or antiestrogen in an antiestrogen-resistant MCF-7 derivative. The p21 levels in the antiestrogen resistant cells increased when c-Myc expression was suppressed, suggesting that loss of p21 regulation was a consequence of constitutive c-Myc expression. Together, these studies implicate p21(WAF1/CIP1) as an important target of c-Myc in breast cancer cells and provide a link between estrogen, c-Myc, and the cell cycle machinery. They further suggest that aberrant c-Myc expression, which is frequently observed in human breast cancers, can contribute to antiestrogen resistance by altering p21(WAF1/CIP1) regulation. PMID- 15757890 TI - Inactivation of Dnmt3b in mouse embryonic fibroblasts results in DNA hypomethylation, chromosomal instability, and spontaneous immortalization. AB - DNA hypomethylation is a hallmark of many types of solid tumors. However, it remains elusive how DNA hypomethylation may contribute to tumorigenesis. In this study, we have investigated how targeted disruption of the DNA methyltransferases Dnmt3a and Dnmt3b affects the growth of mouse embryonic fibroblasts (MEFs). Our studies led to the following observations. 1) Constitutive or conditional deletion of Dnmt3b, but not Dnmt3a, resulted in partial loss of DNA methylation throughout the genome, suggesting that Dnmt3b, in addition to the major maintenance methyltransferase Dnmt1, is required for maintaining DNA methylation in MEF cells. 2) Dnmt3b-deficient MEF cells showed aneuploidy and polyploidy, chromosomal breaks, and fusions. 3) Inactivation of Dnmt3b resulted in either premature senescence or spontaneous immortalization of MEF cells. 4) The G(1) to S-phase checkpoint was intact in primary and spontaneously immortalized Dnmt3b deficient MEFs because the p53 protein was inducible by DNA damage. Interestingly, protein levels of the cyclindependent kinase inhibitor p21 were increased in immortalized Dnmt3b-deficient MEFs even in the absence of p53 induction. These results suggest that DNA hypomethylation may induce genomic instability, which in turn leads to spontaneous immortalization or premature senescence of Dnmt3b-deficient MEFs via a p53-independent mechanism. PMID- 15757891 TI - Transformation by oncogenic RAS sensitizes human colon cells to TRAIL-induced apoptosis by up-regulating death receptor 4 and death receptor 5 through a MEK dependent pathway. AB - RAS oncogenes play a major role in cancer development by activating an array of signaling pathways, most notably mitogen-activated protein kinases, resulting in aberrant proliferation and inhibition of apoptotic signaling cascades, rendering transformed cells resistant to extrinsic death stimuli. However, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is able to kill specific tumor cells through the engagement of its receptors, death receptor 4 (DR4) and death receptor 5 (DR5), and the activation of apoptotic pathways, providing promising targets for anticancer therapies. In this study, we show that TRAIL induces cell death in human colon adenocarcinoma cells in a MEK-dependent manner. We also report a prolonged MEK-dependent activation of ERK1/2 and increased c-FOS expression induced by TRAIL in this system. Our study reveals that transformation of the colon cell line Caco-2 by Ki- and mainly by Ha-ras oncogenes sensitizes these cells to TRAIL-induced apoptosis by causing specific MEK-dependent up regulation of DR4 and DR5. These observations taken together reveal that RAS-MEK ERK1/2 signaling pathway can sensitize cells to TRAIL-induced apoptosis by up regulating DR4 and DR5 and overall imply that TRAIL-based therapeutic strategies using TRAIL agonists could be used in cases of human colon cancers bearing RAS mutations. PMID- 15757892 TI - The average conformation at micromolar [Ca2+] of Ca2+-atpase with bound nucleotide differs from that adopted with the transition state analog ADP.AlFx or with AMPPCP under crystallization conditions at millimolar [Ca2+]. AB - Crystalline forms of detergent-solubilized sarcoplasmic reticulum Ca2+-ATPase, obtained in the presence of either a substrate analog, AMPPCP, or a transition state complex, ADP.fluoroaluminate, were recently described to share the same general architecture despite the fact that, when studied in a test tube, these forms show different functional properties. Here, we show that the differences in the properties of the E1.AMPPCP and the E1.ADP.AlFx membraneous (or solubilized) forms are much less pronounced when these properties are examined in the presence of 10 mM Ca2+ (the concentration prevailing in the crystallization media) than when they are examined in the presence of the few micromolar of Ca2+ known to be sufficient to saturate the transport sites. This concerns various properties, including ATPase susceptibility to proteolytic cleavage by proteinase K, ATPase reactivity toward SH-directed Ellman's reagent, ATPase intrinsic fluorescence properties (here described for the E1.ADP.AlFx complex for the first time), and also the rates of 45Ca2+-40Ca2+ exchange at site "II." These results solve the above paradox at least partially and suggest that the presence of a previously unrecognized Ca2+ ion in the E1.AMPPCP crystals should be re-investigated. A contrario, they emphasize the fact that the average conformation of the E1.AMPPCP complex under usual conditions in the test tube differs from that found in the crystalline form. The extended conformation of nucleotide revealed by the E1.AMPPCP crystalline form might be only indicative of the requirements for further processing of the complex, toward the transition state leading to phosphorylation and Ca2+ occlusion. PMID- 15757893 TI - A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity. AB - Both the epidermal growth factor receptor (EGFR) and the insulin-like growth factor receptor (IGFR) have been implicated in the tumorigenesis of a variety of cancers. Here we propose that simultaneous targeting of both receptors with a bispecific antibody would lead to enhanced antitumor activity. To this end, we produced a recombinant human IgG-like bispecific antibody, a Di-diabody, using the variable regions from two antagonistic antibodies: IMC-11F8 to EGFR and IMC A12 to IGFR. The Di-diabody binds to both EGFR and IGFR and effectively blocked both EGF- and IGF-stimulated receptor activation and tumor cell proliferation. The Di-diabody also inherited the biological properties from both of its parent antibodies; it triggers rapid and significant IGFR internalization and degradation and mediates effective antibody-dependent cellular cytotoxicity in a variety of tumor cells. Finally, the Di-diabody strongly inhibited the growth of two different human tumor xenografts in vivo. Our results underscore the benefits of simultaneous targeting of two tumor targets with bispecific antibodies. PMID- 15757895 TI - Complex formation between glutamyl-tRNA reductase and glutamate-1-semialdehyde 2,1-aminomutase in Escherichia coli during the initial reactions of porphyrin biosynthesis. AB - In Escherichia coli the first common precursor of all tetrapyrroles, 5 aminolevulinic acid, is synthesized from glutamyl-tRNA (Glu-tRNA(Glu)) in a two step reaction catalyzed by glutamyl-tRNA reductase (GluTR) and glutamate-1 semialdehyde 2,1-aminomutase (GSA-AM). To protect the highly reactive reaction intermediate glutamate-1-semialdehyde (GSA), a tight complex between these two enzymes was proposed based on their solved crystal structures. The existence of this hypothetical complex was verified by two independent biochemical techniques. Co-immunoprecipitation experiments using antibodies directed against E. coli GluTR and GSA-AM demonstrated the physical interaction of both enzymes in E. coli cell-free extracts and between the recombinant purified enzymes. Additionally, the formation of a GluTR.GSA-AM complex was identified by gel permeation chromatography. Complex formation was found independent of Glu-tRNA(Glu) and cofactors. The analysis of a GluTR mutant truncated in the 80-amino acid C terminal dimerization domain (GluTR-A338Stop) revealed the importance of GluTR dimerization for complex formation. The in silico model of the E. coli GluTR.GSA AM complex suggested direct metabolic channeling between both enzymes to protect the reactive aldehyde species GSA. In accordance with this proposal, side product formation catalyzed by GluTR was observed via high performance liquid chromatography analysis in the absence of the GluTR.GSA-AM complex. PMID- 15757896 TI - The global transcriptional response of Escherichia coli to induced sigma 32 protein involves sigma 32 regulon activation followed by inactivation and degradation of sigma 32 in vivo. AB - sigma(32) is the first alternative sigma factor discovered in Escherichia coli and can direct transcription of many genes in response to heat shock stress. To define the physiological role of sigma(32), we have used transcription profiling experiments to identify, on a genome-wide basis, genes under the control of sigma(32) in E. coli by moderate induction of a plasmid-borne rpoH gene under defined, steady-state growth conditions. Together with a bioinformatics approach, we successfully confirmed genes known previously to be directly under the control of sigma(32) and also assigned many additional genes to the sigma(32) regulon. In addition, to understand better the functional relevance of the increased amount of sigma(32) to changes in the transcriptional level of sigma(32)-dependent genes, we measured the protein level of sigma(32) both before and after induction by a newly developed quantitative Western blot method. At a normal constant growth temperature (37 degrees C), we found that the sigma(32) protein level rapidly increased, plateaued, and then gradually decreased after induction, indicating sigma(32) can be regulated by genes in its regulon and that the mechanisms of sigma(32) synthesis, inactivation, and degradation are not strictly temperature-dependent. The decrease in the transcriptional level of sigma(32) dependent genes occurs earlier than the decrease in full-length sigma(32) in the wild type strain, and the decrease in the transcriptional level of sigma(32) dependent genes is greatly diminished in a DeltaDnaK strain, suggesting that DnaK can act as an anti-sigma factor to functionally inactivate sigma(32) and thus reduce sigma(32)-dependent transcription in vivo. PMID- 15757894 TI - Persistent nuclear factor-kappa B activation in Ucp2-/- mice leads to enhanced nitric oxide and inflammatory cytokine production. AB - One of the phenotypes of mice with targeted disruption of the uncoupling protein 2 gene (Ucp2-/-) is greater macrophage phagocytic activity and free radical production, resulting in a striking resistance to infectious microorganisms. In this study, the molecular mechanisms of this enhanced immune response were investigated. We found that levels of nitric oxide measured in either plasma or isolated macrophages from Ucp2-/- mice are significantly elevated in response to bacterial lipopolysaccharide challenge compared with similarly treated Ucp2+/+ mice. Likewise, expression of inducible nitric-oxide synthase and inflammatory cytokines is higher in Ucp2-/- mice in vivo and in vitro. Key steps in the activation cascade of nuclear factor (NF)-kappa B, including I kappa B kinase and nuclear translocation of NF-kappa B subunits, are all remarkably enhanced in Ucp2 /- mice, most notably even under basal conditions. The elevated basal activity of I kappa B kinase in macrophages from Ucp2-/- mice can be blocked by cell permeable inhibitors of superoxide and hydrogen peroxide generation, but not by a specific inhibitor for inducible nitric-oxide synthase. Isolated mitochondria from Ucp2-/- cells produced more superoxide/hydrogen peroxide. We conclude that mitochrondrially derived reactive oxygen from Ucp2-/- cells constitutively activates NF-kappa B, resulting in a "primed" state to both potentiate and amplify the inflammatory response upon subsequent stimulation. PMID- 15757897 TI - MxA, a member of the dynamin superfamily, interacts with the ankyrin-like repeat domain of TRPC. AB - Mammalian transient receptor potential canonical channels have been proposed as the molecular entities associated with calcium entry activity in nonexcitable cells. Amino acid sequence analyses of TRPCs revealed the presence of ankyrin like repeat domains, one of the most common protein-protein interaction motifs. Using a yeast two-hybrid interaction assay, we found that the second ankyrin-like repeat domain of TRPC6 interacted with MxA, a member of the dynamin superfamily. Using a GST pull-down and co-immunoprecipitation assay, we showed that MxA interacted with TRPC1, -3, -4, -5, -6, and -7. Overexpression of MxA in HEK293T cells slightly increased endogenous calcium entry subsequent to stimulation of G(q) protein-coupled receptors or store depletion by thapsigargin. Co-expression of MxA with TRPC6 enhanced agonist-induced or OAG-induced calcium entry activity. GTP binding-defective MxA mutants had only a minor potentiating effect on OAG induced TRPC6 activity. However, a MxA mutant that could bind GTP but that lacked GTPase activity produced the same effect as MxA on OAG-induced TRPC6 activity. These results indicated that MxA interacted specifically with the second ankyrin like repeat domain of TRPCs and suggested that monomeric MxA regulated the activity of TRPC6 by a mechanism requiring GTP binding. Additional results showed that an increase in the endogenous expression of MxA, induced by a treatment with interferon alpha, regulated the activity of TRPC6. The study clearly identified MxA as a new regulatory protein involved in Ca2+ signaling. PMID- 15757898 TI - The MRE11-RAD50-XRS2 complex, in addition to other non-homologous end-joining factors, is required for V(D)J joining in yeast. AB - Lymphoid cells of the vertebrate immune system rely on factors in the non homologous end-joining (NHEJ) DNA repair pathway to form signal joints during V(D)J recombination. Unlike other end-joining reactions, signal joint formation is a specialized case of NHEJ that also requires the lymphoid-specific RAG proteins. Whether V(D)J recombination requires the Mre11-Rad50-Nbs1 complex remains an open question, as null mutations in any member of the complex are lethal in mammals. However, Saccharomyces cerevisiae strains carrying null mutations in components of the homologous Mre11p-Rad50p-Xrs2p (MRX) complex are viable. We therefore took advantage of a recently developed V(D)J recombination assay in yeast to assess the role of MRX in V(D)J joining. Here we confirmed that signal joint formation in yeast is dependent on the same NHEJ factors known to be required in mammalian cells. In addition, we showed an absolute requirement for the MRX complex in signal joining, suggesting that the Mre11-Rad50-Nbs1 complex may be required for signal joint formation in mammalian cells as well. PMID- 15757899 TI - Protein kinase C-dependent regulation of NAG-1/placental bone morphogenic protein/MIC-1 expression in LNCaP prostate carcinoma cells. AB - NAG-1 (nonsteroidal anti-inflammatory drug-activated gene), a member of the transforming growth factor beta superfamily, is involved in cellular processes such as inflammation, apoptosis/survival, and tumorigenesis and is regulated by p53, Sp1, and Egr-1. In the current study, the regulation of NAG-1 expression in LNCaP human prostate carcinoma cells by 12-O-tetradecanoylphorbol-13-acetate (TPA) was examined. TPA treatment increased NAG-1 protein and mRNA levels in a time- and concentration-dependent manner as well as NF-kappa B binding/transcriptional activity in LNCaP cells. Pretreatment with protein kinase C (PKC) inhibitor blocked the TPA-induced increase in NAG-1 protein levels and NF kappa B binding/transcriptional activity, whereas an inhibition of p38, JNK, MEK activity had no effect on TPA-induced NAG-1 levels and NF-kappa B transcriptional activity. Expression of constitutively active PKCs induced an increase in NF kappa B transcriptional activity and NAG-1 protein levels in LNCaP cells. The expression of NF-kappa B p65 induced NAG-1 promoter activity, and chromatin immunoprecipitation assay for p65 showed that NF-kappa B binds the NAG-1 promoter in LNCaP cells. Inhibition of TPA-induced NAG-1 expression by NAG-1 short interfering RNA blocked TPA-induced apoptosis in LNCaP cells, suggesting induction of NAG-1 negatively affects LNCaP cell survival. These results demonstrate that NAG-1 expression is up-regulated by TPA in LNCaP cells through a PKC-dependent pathway involving the activation of NF-kappa B. PMID- 15757900 TI - JNK1 differentially regulates osteopontin-induced nuclear factor-inducing kinase/MEKK1-dependent activating protein-1-mediated promatrix metalloproteinase 9 activation. AB - We have recently demonstrated that nuclear factor-inducing kinase (NIK) plays a crucial role in osteopontin (OPN)-induced mitogen-activated protein kinase/I kappa B alpha kinase-dependent nuclear factor kappa B (NF kappa B)-mediated promatrix metalloproteinase-9 activation (Rangaswami, H., Bulbule, A., and Kundu, G. C. (2004) J. Biol. Chem. 279, 38921-38935). However, the molecular mechanism(s) by which OPN regulates NIK/MEKK1-dependent activating protein-1 (AP 1)-mediated promatrix metalloproteinase-9 activation and whether JNK1 plays any role in regulating both these pathways that control the cell motility are not well defined. Here we report that OPN induces alpha v beta3 integrin-mediated MEKK1 phosphorylation and MEKK1-dependent JNK1 phosphorylation and activation. Overexpression of NIK enhances OPN-induced c-Jun expression, whereas overexpressed NIK had no role in OPN-induced JNK1 phosphorylation and activation. Sustained activation of JNK1 by overexpression of wild type but not kinase negative MEKK1 resulted in suppression of ERK1/2 activation. But this did not affect the OPN-induced NIK-dependent ERK1/2 activation. OPN stimulated both NIK and MEKK1-dependent c-Jun expression, leading to AP-1 activation, whereas NIK dependent AP-1 activation is independent of JNK1. OPN also enhanced JNK1 dependent/independent AP-1-mediated urokinase type plasminogen activator (uPA) secretion, uPA-dependent promatrix metalloproteinase-9 (MMP-9) activation, cell motility, and invasion. OPN stimulates tumor growth, and the levels of c-Jun, AP 1, urokinase type plasminogen activator, and MMP-9 were higher in OPN-induced tumor compared with control. To our knowledge this is first report that OPN induces NIK/MEKK1-mediated JNK1-dependent/independent AP-1-mediated pro-MMP-9 activation and regulates the negative crosstalk between NIK/ERK1/2 and MEKK1/JNK1 pathways that ultimately controls the cell motility, invasiveness, and tumor growth. PMID- 15757901 TI - Thermal destabilization of non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Streptococcus mutans upon phosphate binding in the active site. AB - Catalysis by the NADP-dependent non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) from Streptococcus mutans, a member of the aldehyde dehydrogenase (ALDH) family, relies on a local conformational reorganization of the active site. This rearrangement is promoted by the binding of NADP and is strongly kinetically favored by the formation of the ternary complex enzyme.NADP.substrate. Adiabatic differential scanning calorimetry was used to investigate the effect of ligands on the irreversible thermal denaturation of GAPN. We showed that phosphate binds to GAPN, resulting in the formation of a GAPN.phosphate binary complex characterized by a strongly decreased thermal stability, with a difference of at least 15 degrees C between the maximum temperatures of the thermal transition peaks. The kinetics of phosphate association and dissociation are slow, allowing both free and GAPN.phosphate complexes to be observed by differential scanning calorimetry and to be separated by native polyacrylamide electrophoresis run in phosphate buffer. Analysis of a set of mutants of GAPN strongly suggests that phosphate is bound to the substrate C-3 subsite. In addition, the substrate analog glycerol-3-phosphate has similar effects as does phosphate on the thermal behavior of GAPN. Based on the current knowledge on the catalytic mechanism of GAPN and other ALDHs, we propose that ligand-induced thermal destabilization is a mechanism that provides to ALDHs the required flexibility for an efficient catalysis. PMID- 15757902 TI - A conserved N-capping motif contributes significantly to the stabilization and dynamics of the C-terminal region of class Alpha glutathione S-transferases. AB - Helix 9, the major structural element in the C-terminal region of class Alpha glutathione transferases, forms part of the active site of these enzymes where its dynamic properties modulate both catalytic and ligandin functions. A conserved aspartic acid N-capping motif for helix 9 was identified by sequence alignments of the C-terminal regions of class Alpha glutathione S-transferases (GSTs) and an analysis by the helix-coil algorithm AGADIR. The contribution of the N-capping motif to the stability and dynamics of the region was investigated by replacing the N-cap residue Asp-209 with a glycine in human glutathione S transferase A1-1 (hGST A1-1) and in a peptide corresponding to its C-terminal region. Far-UV circular dichroism and AGADIR analyses indicate that, in the absence of tertiary interactions, the wild-type peptide displays a low intrinsic tendency to form a helix and that this tendency is reduced significantly by the Asp-to-Gly mutation. Disruption of the N-capping motif of helix 9 in hGST A1-1 alters the conformational dynamics of the C-terminal region and, consequently, the features of the H-site to which hydrophobic substrates (e.g. 1-chloro-2,4 dinitrobenzene (CDNB)) and nonsubstrates (e.g. 8-anilino-1-naphthalene sulfonate (ANS)) bind. Isothermal calorimetric and fluorescence data for complex formation between ANS and protein suggest that the D209G-induced perturbation in the C terminal region prevents normal ligand-induced localization of the region at the active site, resulting in a less hydrophobic and more solvent-exposed H-site. Therefore, the catalytic efficiency of the enzyme with CDNB is diminished due to a lowered affinity for the electrophilic substrate and a lower stabilization of the transition state. PMID- 15757903 TI - Identification of transcriptional targets of HOXA5. AB - The homeobox gene HOXA5 encodes a transcription factor that has been shown to play important roles in embryogenesis, hematopoiesis, and tumorigenesis. In order to decipher downstream signaling pathways of HOXA5, we utilized oligonucleotide microarray analysis to identify genes that are differentially expressed in HOXA5 induced cells compared with uninduced cells. Comparative analysis of gene expression changes after 9 h of HOXA5 induction in Hs578T breast cancer cells identified 306 genes whose expression was modulated at least 2-fold. Ten of these 306 genes were also up-regulated by at least 2-fold at 6 h post-induction. The expression of all of these 10 genes was confirmed by semiquantitative reverse transcription-PCR. Among these 10 genes, which are most likely to be direct targets of HOXA5, we initiated an investigation into the pleiotrophin gene by first cloning its promoter. Transient transfection assays indicated that HOXA5 can specifically activate the pleiotrophin promoter. Promoter deletion, chromatin immunoprecipitation assay, and gel-shift assays were performed to show that HOXA5 can directly bind to one binding site on the pleiotrophin promoter. These data strongly suggest that microarray analysis can successfully identify many potential direct downstream genes of HOXA5. Further functional analysis of these targets will allow us to better understand the diverse functions of HOXA5 in embryonic development and tumorigenesis. PMID- 15757904 TI - Substrate binding stoichiometry and kinetics of the norepinephrine transporter. AB - The human norepinephrine (NE) transporter (hNET) attenuates neuronal signaling by rapid NE clearance from the synaptic cleft, and NET is a target for cocaine and amphetamines as well as therapeutics for depression, obsessive-compulsive disorder, and post-traumatic stress disorder. In spite of its central importance in the nervous system, little is known about how NET substrates, such as NE, 1 methyl-4-tetrahydropyridinium (MPP+), or amphetamine, interact with NET at the molecular level. Nor do we understand the mechanisms behind the transport rate. Previously we introduced a fluorescent substrate similar to MPP+, which allowed separate and simultaneous binding and transport measurement (Schwartz, J. W., Blakely, R. D., and DeFelice, L. J. (2003) J. Biol. Chem. 278, 9768-9777). Here we use this substrate, 4-(4-(dimethylamino)styrl)-N-methyl-pyridinium (ASP+), in combination with green fluorescent protein-tagged hNETs to measure substrate transporter stoichiometry and substrate binding kinetics. Calibrated confocal microscopy and fluorescence correlation spectroscopy reveal that hNETs, which are homomultimers, bind one substrate molecule per transporter subunit. Substrate residence at the transporter, obtained from rapid on-off kinetics revealed in fluorescence correlation spectroscopy, is 526 micros. Substrate residence obtained by infinite dilution is 1000 times slower. This novel examination of substrate-transporter kinetics indicates that a single ASP+ molecule binds and unbinds thousands of times before being transported or ultimately dissociated from hNET. Calibrated fluorescent images combined with mass spectroscopy give a transport rate of 0.06 ASP+/hNET-protein/s, thus 36,000 on-off binding events (and 36 actual departures) occur for one transport event. Therefore binding has a low probability of resulting in transport. We interpret these data to mean that inefficient binding could contribute to slow transport rates. PMID- 15757905 TI - Mechanisms involved in enhancement of osteoclast formation and function by low molecular weight hyaluronic acid. AB - Hyaluronic acid (HA) is a component of the extracellular matrix that has been shown to play an important role in bone formation, resorption, and mineralization both in vivo and in vitro. We examined the effects of HA at several molecular weights on osteoclast formation and function induced by RANKL (receptor activator of NF-kappa B ligand) in a mouse monocyte cell line (RAW 264.7). HA at M(r) < 8,000 (low molecular weight HA (LMW-HA)) enhanced tartrate-resistant acid phosphatase-positive multinucleated cell formation and tartrate-resistant acid phosphatase activity induced by RANKL in a dose-dependent manner, whereas HA at M(r) > 900,000 (high molecular weight HA (HMW-HA)) showed no effect on osteoclast differentiation. LMW-HA enhanced pit formation induced by RAW 264.7 cells, whereas HMW-HA did not, and LMW-HA stimulated the expression of RANK (receptor activator of NF-kappa B) protein in RAW 264.7 cells. In addition, we found that LMW-HA enhanced the levels of c-Src protein and phosphorylation of ERKs and p38 MAPK in RAW 264.7 cells stimulated with RANKL, whereas the p38 MAPK inhibitor SB203580 inhibited RANKL-induced osteoclast differentiation. This enhancement of c-Src and RANK proteins induced by LMW-HA was inhibited by CD44 function-blocking monoclonal antibody. These results indicate that LMW-HA plays an important role in osteoclast differentiation and function through the interaction of RANKL and RANK. PMID- 15757906 TI - Identification and functional characterization of a novel low affinity aromatic preferring amino acid transporter (arpAT). One of the few proteins silenced during primate evolution. AB - We have identified in silico arpAT, a gene encoding a new member of the LSHAT family, and cloned it from kidney. Co-expression of arpAT with the heavy subunits rBAT or 4F2hc elicited a sodium-independent alanine transport activity in HeLa cells. L-tyrosine, l-3,4-dihydroxyphenylalanine (L-DOPA), L-glutamine, L-serine, L-cystine, and L-arginine were also transported. Kinetic and cis-inhibition studies showed a K(m) = 1.59 +/- 0.24 mM for L-alanine or IC50 in the millimolar range for most amino acids, except L-proline, glycine, anionic and D-amino acids, which were not inhibitory. L-DOPA and L-tyrosine were the most effective competitive inhibitors of L-alanine transport, with IC50 values of 272.2 +/- 57.1 and 716.3 +/- 112.4 microM, respectively. In the small intestine, arpAT mRNA was located at the enterocytes, in a decreasing gradient from the crypts to the tip of the villi. It was also expressed in neurons from different brain areas. Finally, we show that while the arpAT gene is conserved in rat, dog, and chicken, it has become silenced in humans and chimpanzee. Actually, it has been recently reported that it is one of the 33 recently inactivated genes in the human lineage. The evolutionary implications of the silencing process and the roles of arpAT in transport of L-DOPA in the brain and in aromatic amino acid absorption are discussed. PMID- 15757907 TI - Fluorescence resonance energy transfer analysis of the antagonist- and partial agonist-occupied states of the cholecystokinin receptor. AB - Changes in receptor conformation are believed to be key for ligand-induced regulation of cellular signaling cascades. However, little information exists about specific conformations of a receptor. We recently applied fluorescence resonance energy transfer to determine distances from distinct points distributed over the surface and within the helical bundle of the cholecystokinin receptor to the amino terminus of a full agonist CCK analogue (Harikumar, K. G., Pinon, D. I., Wessels, W. S., Dawson, E. S., Lybrand, T. P., Prendergast, F. G., and Miller, L. J. (2004) Mol. Pharmacol. 65, 28-35). Here, we apply the same experimental strategy to determine distances from the same receptor positions to an analogous point at the amino terminus of structurally related partial agonist (Alexa488-Gly-[(Nle(28,31))CCK-26-32]phenethyl ester) and antagonist (Alexa488 Gly-[(D-Trp31, Nle(28,31))CCK-26-32]phenethyl ester) ligands. A high degree of spectral overlap and fluorescence transfer was observed for ligand-occupied fluorescent-tagged receptors with no transfer observed for the ligand-occupied pseudo-wild type null cysteine-reactive mutant receptor (C94S). For the partial agonist, calculated distances to receptor positions 94, 102, 204, and 341, representing sites within the helical confluence, and the first, second, and third loops, were 21 +/- 0.4, 18 +/- 0.4, 25 +/- 1, and 17 +/- 1 angstroms, not different from those measured previously for the analogous full agonist. For the antagonist, the analogous distances were 21 +/- 2, 28 +/- 2, 15 +/- 1 and 21 +/- 1 angstroms. Distances to the first and third loops were longer and the distance to the second loop was shorter for the antagonist relative to both the full and partial agonist probes, whereas all three probes demonstrated similar distances to the intrahelical reference point. This supports the possibilities of changes in the conformation of the probe and/or the receptor induced by structurally similar ligands having distinct intrinsic biological activities. PMID- 15757908 TI - Integrin alpha5/beta1 expression mediates HER-2 down-regulation in colon cancer cells. AB - HER-2 is constitutively activated and overexpressed in many cancers, and its inhibition in colon cancer cells diminishes tumorigenicity and induces apoptosis. Little is known about the regulation of HER-2 signaling in colon cancer cells. Integrin alpha5/beta1 expression is frequently lost in colorectal cancer cells compared with normal intestinal epithelium, and colon cancer cells lacking integrin alpha5/beta1 expression utilize HER-2 signaling for proliferation and tumorigenicity. Re-expression of integrin alpha5/beta1 in colon cancer cells abrogated their tumorigenicity, but how this occurs is not well known. Stable expression of integrin alpha5/beta1 in colon cancer cells with little or no detectable integrin alpha5/beta1 protein expression resulted in the post transcriptional down-regulation of HER-2 protein. Integrin alpha5/beta1 was found to interact with HER-2, and the cytoplasmic domain of integrin alpha5/beta1 was sufficient to mediate HER-2 down-regulation. Integrin alpha5/beta1-mediated down regulation of HER-2 was the result of increased lysosomal targeting. The inhibition of HER-2 signaling represents a potential mechanism by which integrin alpha5/beta1 exerts its tumor suppressor-like activity in colon cancer cells. These results also suggest that a novel function for integrin alpha5/beta1 is the control of HER-2 expression. PMID- 15757909 TI - Assembly of the kappa preB receptor requires a V kappa-like protein encoded by a germline transcript. AB - By confining germline transcription as a byproduct of the mechanisms inherent to genetic rearrangements, the translation of respective mRNAs and their biological relevance might have been overlooked. Here we report the identification, cloning, and biochemical characterization of a human Vkappa-like protein that is encoded by a germline transcript. This surrogate protein assembles with the immunoglobulin mu heavy chain at the surface of B cell progenitors and precursors to form a kappa-like antigen receptor. These findings support the notion that germline transcription is not futile and stress the flexibility in eukaryotic gene usage and expression. In addition, the present study confirms the co existence of surrogate lambda and kappa receptors that are proposed to work in concert to promote B lymphocyte maturation. PMID- 15757910 TI - Bax-dependent regulation of Bak by voltage-dependent anion channel 2. AB - Many studies have demonstrated a critical role of Bax in mediating apoptosis, but the role of Bak in regulating cancer cell apoptotic sensitivities in the presence or absence of Bax remains incompletely understood. Using isogenic cells with defined genetic deficiencies, here we show that in response to intrinsic, extrinsic, and endoplasmic reticulum stress stimuli, HCT116 cells show clear-cut apoptotic sensitivities in the order of Bax+/Bak+ > Bax+/Bak- >> Bax-/Bak+ >> Bax /Bak-. Small interference RNA-mediated knockdown of Bak in Bax-deficient cells renders HCT116 cells completely resistant to apoptosis induction. Surprisingly, however, Bak knockdown in Bax-expressing cells only slightly affects the apoptotic sensitivities. Bak, like Bax, undergoes the N terminus exposure upon apoptotic stimulation in both Bax-expressing and Bax-deficient cells. Gel filtration, chemical cross-linking, and co-immunoprecipitation experiments reveal that different from Bax, which normally exists as monomers in unstimulated cells and is oligomerized by apoptotic stimulation, most Bak in unstimulated HCT116 cells exists in two distinct protein complexes, one of which contains voltage dependent anion channel (VDAC) 2. During apoptosis, Bak and Bax form both homo- and hetero-oligomeric complexes that still retain some VDAC-2. However, the oligomeric VDAC-2 complexes are diminished, and Bak does not interact with VDAC-2 in Bax-deficient HCT116 cells. These results highlight VDAC-2 as a critical inhibitor of Bak-mediated apoptotic responses. PMID- 15757911 TI - Identification by site-directed mutagenesis and chemical modification of three vicinal cysteine residues in rat mitochondrial carnitine/acylcarnitine transporter. AB - The proximity of the Cys residues present in the mitochondrial rat carnitine/acylcarnitine carrier (CAC) primary structure was studied by using site directed mutagenesis in combination with chemical modification. CAC mutants, in which one or more Cys residues had been replaced with Ser, were overexpressed in Escherichia coli and reconstituted into liposomes. The effect of SH oxidizing, cross-linking, and coordinating reagents was evaluated on the carnitine/carnitine exchange catalyzed by the recombinant reconstituted CAC proteins. All the tested reagents efficiently inhibited the wild-type CAC. The inhibitory effect of diamide, Cu(2+)-phenanthroline, or phenylarsine oxide was largely reduced or abolished by the double substitutions C136S/C155S, C58S/C136S, and C58S/C155S. The decrease in sensitivity to these reagents was much lower in double mutants in which Cys(23) was substituted with Cys(136) or Cys(155). No decrease in inhibition was found when Cys(89) and/or Cys(283) were replaced with Ser. Sb(3+), which coordinates three cysteines, inhibited only the Cys replacement mutants containing cysteines 58, 136, and 155 of the six native cysteines. In addition, the mutant C23S/C89S/C155S/C283S, in which double tandem fXa recognition sites were inserted in positions 65-72, i.e. between Cys(58) and Cys(136), was not cleaved into two fragments by fXa protease after treatment with diamide. These results are interpreted in light of the homology model of CAC based on the available x-ray structure of the ADP/ATP carrier. They indicate that Cys(58), Cys(136), and Cys(155) become close in the tertiary structure of the CAC during its catalytic cycle. PMID- 15757914 TI - Racial and ethnic health disparities and the unfinished civil rights agenda. AB - Civil rights-era efforts to end disparities in health care in federally financed health programs faced three successively more difficult challenges: (1) ending Jim Crow practices, (2) eliminating more subtle forms of segregation, and (3) assuring nondiscriminatory treatment in integrated settings. Federal efforts peaked with the implementation of the Medicare program. Visible symbols of Jim Crow disappeared, and most crude disparities in access were eliminated. The unfinished parts of the civil rights-era agenda, the persistence of more subtle forms of segregation, and the failure to assure nondiscriminatory treatment pose major challenges to current efforts to eliminate health care disparities. PMID- 15757915 TI - Social sources of racial disparities in health. AB - Racial disparities in mortality over time reflect divergent pathways to the current large racial disparities in health. The residential concentration of African Americans is high and distinctive, and the related inequities in neighborhood environments, socioeconomic circumstances, and medical care are important factors in initiating and maintaining racial disparities in health. Efforts are needed to identify and maximize health-enhancing resources that may reduce some of the negative effects of psychosocial factors on health. Health and health disparities are embedded in larger historical, geographic, sociocultural, economic, and political contexts. Changes in a broad range of public policies are likely to be central to effectively addressing racial disparities. PMID- 15757916 TI - Policy challenges in addressing racial disparities and improving population health. AB - Socioeconomic status fundamentally affects most health and disease outcomes, but black Americans are doubly disadvantaged by low status, discrimination, and residential segregation. Improving health and removing disparities are essential goals, but some efforts that improve the health of blacks in important ways also increase black-white disparity ratios. People with more information, influence, resources, and social networks may be better able to take advantage of new technologies and scientific developments, initially increasing disparities. Better health and reduced mortality should be the key policy criteria, but these criteria should be linked with consideration of careful targeting to level the playing field and close disparities. PMID- 15757917 TI - Communities and health policy: a pathway for change. AB - Improving the health system can reduce the effects of health disparities, but it can do little to eliminate them. An upsurge in new research is documenting the impact of physical, social, and economic environmental factors: air quality, housing conditions, racism, relationship to community institutions, and neighborhood economic conditions, all of which affect health status over time. A combined focus on community and the policies that affect communities' environments presents opportunities for altering and ameliorating the underlying forces at the heart of the determinants of health. This Perspective presents examples of successful community involvement and policy change. PMID- 15757918 TI - Health disparities by race and class: why both matter. AB - In this essay we examine three competing causal interpretations of racial disparities in health. The first approach views race as a biologically meaningful category and racial disparities in health as reflecting inherited susceptibility to disease. The second approach treats race as a proxy for class and views socioeconomic stratification as the real culprit behind racial disparities. The third approach treats race as neither a biological category nor a proxy for class, but as a distinct construct, akin to caste. We point to historical, political, and ideological obstacles that have hindered the analysis of race and class as codeterminants of disparities in health. PMID- 15757919 TI - Disparities and quality improvement: federal policy levers. AB - Using a quality improvement framework to address racial and ethnic disparities in health care highlights multiple opportunities for federal and state governments to exert policy leverage, particularly through their roles as purchasers and regulators. Under such a framework, federal and state governments can expand their roles in collecting race/ethnicity data; define universal and meaningful race/ethnicity categories; more broadly disseminate standards for cultural competence; and demand the reduction of disparities through leveraging their status as collectively the largest U.S. health care payer. PMID- 15757920 TI - Medicare as a catalyst for reducing health disparities. AB - As the nation's largest purchaser and regulator of health care, Medicare is positioned to be a leader in reducing racial and ethnic health disparities. Its leverage was demonstrated in 1966-the year of Medicare's inception-when hospitals desegregated as a condition for receiving Medicare reimbursement. Since then, Medicare has contributed to dramatic improvement in the health of the elderly and disabled minority population, although disparities between minority and white beneficiaries remain. A National Academy of Social Insurance study panel is exploring how Medicare could use its leverage to reduce disparities, for both its beneficiaries and the rest of the nation. PMID- 15757921 TI - Compiling the evidence: the National Healthcare Disparities Reports. AB - Disparities in health care have been described extensively in the literature. The next step in resolving this national problem is to develop the necessary infrastructure for monitoring and tracking disparities. The congressionally mandated National Healthcare Disparities Report begins to build this infrastructure. The 2003 report addressed many of the methodological challenges inherent in measuring disparities. The recently released 2004 report continues the process by summarizing the status of U.S. health care disparities and beginning to track changes over time. Both reports emphasize the need to integrate activities to reduce disparities and to improve the quality of health care. PMID- 15757922 TI - Creating a state minority health policy report card. AB - A state minority health policy report card may provide an important tool for evaluating and promoting state policies to reduce health disparities. This study develops criteria that can form the basis of such a state report card and assesses the performance of all fifty states on these measures. The results indicate wide variation among states, with geographic region being a significant predictor of performance on all four measures. Future research should be conducted on other predictors of state variation in minority health policy and connections between state policy and health outcomes for minorities. PMID- 15757923 TI - The role of health insurance coverage in reducing racial/ethnic disparities in health care. AB - Research showing racial/ethnic disparities in medical care obtained by people with comparable insurance has raised questions about the extent to which health insurance improves opportunities for care. To assess whether insurance expansions could be expected to reduce racial/ethnic disparities in access to care, this paper reviews evidence from studies specifically designed to quantify the contribution of health insurance to racial/ethnic disparities in access. The studies provide evidence that a sizable share of the differences in whether a person has a regular source of care could be reduced if Hispanics and African Americans were insured at levels comparable to those of whites. PMID- 15757924 TI - Health care organizations' use of race/ethnicity data to address quality disparities. AB - Health care organizations-health plans, hospitals, community health centers, clinics, and group practices-can play an important role in the elimination of racial/ethnic disparities in health care. There are now a number of examples of organizations that have been successful in reducing or eliminating disparities, and a number of published examples of how quality improvement initiatives can improve care for members of targeted minority groups, thereby contributing to the elimination of disparities. PMID- 15757925 TI - Taking on racial and ethnic disparities in health care: the experience at Aetna. AB - Among the nation's health plans, Aetna is considered the industry leader in efforts to eliminate racial and ethnic disparities in health care. This Perspective describes the work of a task force led by Aetna's chairman and CEO, charged with a number of strategic activities including cultural competency training and the identification of disparities occurring within the Aetna membership population. The cornerstone of this quality-of-care initiative is a successful and ongoing data collection enterprise. Aetna is putting the data to work in its chronic disease management, breast health, and African American Preterm Labor Prevention and Breastfeeding programs. PMID- 15757926 TI - Verizon works to eliminate disparities in health care for its diverse workforce. AB - Eradicating disparities in health care is an ongoing process that must continue through actions from a variety of stakeholders, including employers. Verizon Communications has a diverse workforce of more than 200,000 employees and is committed to attracting and retaining a workforce that mirrors its customers, speaks their languages, and understands their needs. The company developed and implemented a comprehensive plan to help ensure that minority employees and their families receive the health care they need and deserve based on the Institute of Medicine's 2003 report Unequal Treatment. PMID- 15757927 TI - Crossing the language chasm. AB - The quality of communication between patients and clinicians can have a major impact on health outcomes, and limited English proficiency can interfere with effective communication. More than ten million U.S. residents speak English poorly or not at all, constituting a language chasm in the health care system. This paper reviews the evidence on the link between linguistic competence and health care quality and the impact of particular language-assistance strategies. Drawing on the experiences of fourteen health plans that have been at the forefront of linguistic competence efforts, we identify lessons for plans, purchasers, policymakers, and researchers on ways to improve the availability and quality of interpreter services. PMID- 15757928 TI - Pay now or pay later: providing interpreter services in health care. AB - Research amply documents that language barriers impede access to health care, compromise quality of care, and increase the risk of adverse health outcomes among patients with limited English proficiency. Federal civil rights policy obligates health care providers to supply language services, but wide gaps persist because insurers typically do not pay for interpreters, among other reasons. Health care financing policies should reinforce existing medical research and legal policies: Payers, including Medicaid, Medicare, and private insurers, should develop mechanisms to pay for interpretation services for patients who speak limited English. PMID- 15757929 TI - Overcoming disparities in U.S. health care. AB - Disparities in U.S. health care result from a complex mixture of systemic quality and access problems intertwined with historic injury. The many dimensions of health disparities include race, ethnicity, socioeconomic status, and geography. It is critically important for policymakers to define the problem correctly so that our solutions address their intended goal-health security for all regardless of socioeconomic characteristics. Further, U.S. efforts to eliminate disparities must also be part of a broader effort to transform health care and thus must focus, first and foremost, on improving the quality of care delivered to the individual patient. PMID- 15757930 TI - The role of the federal government in eliminating health disparities. AB - Minorities live sicker and die sooner from too many acute and chronic illnesses. To eliminate racial and ethnic disparities in health, Congress and the Bush administration must address the serious challenge of increasing minorities' access to health care and improving the quality of care they receive. We clearly need to expand Medicaid and the State Children's Health Insurance Program (SCHIP), but we must also develop and train culturally competent providers, increase the diversity of the health care workforce, collect better race/ethnicity health data, and make a greater investment in public health. PMID- 15757931 TI - What if we were equal? A comparison of the black-white mortality gap in 1960 and 2000. AB - The United States has made progress in decreasing the black-white gap in civil rights, housing, education, and income since 1960, but health inequalities persist. We examined trends in black-white standardized mortality ratios (SMRs) for each age-sex group from 1960 to 2000. The black-white gap measured by SMR changed very little between 1960 and 2000 and actually worsened for infants and for African American men age thirty-five and older. In contrast, SMR improved in African American women. Using 2002 data, an estimated 83,570 excess deaths each year could be prevented in the United States if this black-white mortality gap could be eliminated. PMID- 15757932 TI - Health center trends, 1994-2001: what do they portend for the federal growth initiative? AB - The Federal Health Center Growth Initiative aims to increase community health centers' (CHCs') capacity by 60 percent from 2002 to 2006. This study investigates how primary care delivery changed and sustained its growth during 1994-2001. Findings reveal a rise in the number of patients and maintenance of their visit rate. People ages 41-64 accounted for the highest percentage of visits in 2001, and continuity of care improved. There were no disparities in visit-based preventive services delivery by race/ethnicity or insurance status. Continued growth under the initiative is likely to help reduce health disparities and improve care for the underserved. PMID- 15757933 TI - The impact of Blue Cross conversions on health spending and the uninsured. AB - Using statewide data on health spending and uninsurance rates, we investigate the impact of Blue Cross conversions on health care costs and coverage. We find mixed results, with some conversion states improving their performance on either or both measures relative to the national average and others experiencing a decline. A multivariate analysis suggests that overall, the impact of Blue Cross conversion may be to reduce hospital and total spending, but whether this effect endures depends in part on how "conversion" is defined. State policymakers and regulators might find these results useful in considering future Blue Cross conversions. PMID- 15757934 TI - Genetic testing for Alzheimer's disease and its impact on insurance purchasing behavior. AB - New genetic tests for adult-onset diseases raise concerns about possible adverse selection in insurance markets. To test for this behavior, we followed 148 cognitively normal people participating in a randomized clinical trial of genetic testing for Alzheimer's disease for one year after risk assessment and Apolipoprotein E (APOE) genotype disclosure. Although no significant differences were found in health, life, or disability insurance purchases, those who tested positive were 5.76 times more likely to have altered their long-term care insurance than those who did not receive APOE genotype disclosure. If genetic testing for Alzheimer's risk assessment becomes common, it could trigger adverse selection in long-term care insurance. PMID- 15757935 TI - Making health equality a reality: the Bronx takes action. AB - In response to growing evidence of racial and ethnic disparities in health, Bronx Health REACH, a coalition of health care providers and community and faith-based organizations, is engaged in an effort to identify and eliminate the root causes of health disparities in their Bronx neighborhood. The group has gained a community perspective on health disparities that it has developed into a seven point advocacy agenda: universal health insurance, an end to segregation in health facilities based on insurance status, accountability for state uncompensated care funds, culturally competent care for all, greater health workforce diversity, an expansion of public health education, and environmental justice. PMID- 15757936 TI - Cultural competence and health care disparities: key perspectives and trends. AB - Cultural competence has gained attention as a potential strategy to improve quality and eliminate racial/ethnic disparities in health care. In 2002 we conducted interviews with experts in cultural competence from managed care, government, and academe to identify their perspectives on the field. We present our findings here and then identify recent trends in cultural competence focusing on health care policy, practice, and education. Our analysis reveals that many health care stakeholders are developing initiatives in cultural competence. Yet the motivations for advancing cultural competence and approaches taken vary depending on mission, goals, and sphere of influence. PMID- 15757937 TI - She walked from El Salvador: caring for an immigrant nation. PMID- 15757938 TI - Concordance: how does a physician who is neither black nor white decide when race is a factor? PMID- 15757939 TI - Use of geocoding in managed care settings to identify quality disparities. AB - Tracking quality-of-care measures is essential for improving care, particularly for vulnerable populations. Although managed care plans routinely track quality measures, few examine whether their performance differs by enrollee race/ethnicity or socioeconomic status (SES), in part because plans do not collect that information. We show that plans can begin examining and targeting potential disparities using indirect measures of enrollee race/ethnicity and SES based on geocoding. Using such measures, we demonstrate disparities within both Medicare+Choice and commercial plans on Health Plan Employer Data and Information Set (HEDIS) measures of diabetes and cardiovascular care, including instances in which race/ethnicity and SES have distinct effects. PMID- 15757940 TI - Medical school tuition and young physicians' indebtedness. AB - Medical school tuition and medical student debt have increased dramatically during the past two decades, but loans are available on favorable terms, which makes it possible for students without personal or family means to get a medical education. As an investment, medical education is an excellent choice; its net present value is more than a million dollars. Cost is nevertheless a strong deterrent to potential applicants, especially minority applicants. If tuition and indebtedness continue to increase while physician incomes do not, there may come a time when only the wealthy can finance a medical education, and medical schools may have increasing difficulty recruiting qualified students. PMID- 15757941 TI - Medicaid policies to contain psychiatric drug costs. AB - This study reviews Medicaid policies to restrict access to psychiatric medications. Policies on prior authorization, preferred drug lists, limitations on the number of prescriptions, fail-first requirements, and use of generics are reviewed. All states apply one or more of those policies to medications for mental illness, and many apply several. A large number of states have legislated exemptions from those policies for certain medications, particularly antipsychotics and antidepressants. Other psychiatric medications are less well protected. Some states appear to restrict access severely. Questions have been raised as to whether these policies actually save money in the long term. PMID- 15757942 TI - Public-private partnerships and antiretroviral drugs for HIV/AIDS: lessons from Botswana. AB - The African Comprehensive HIV/AIDS Partnerships (ACHAP) played a major role in initiating Botswana's antiretroviral (ARV) program in 2001. ACHAP is a prominent public-private partnership involving Merck and its foundation, the Bill and Melinda Gates Foundation, and the government of Botswana. This paper analyzes ACHAP's efforts to assist Botswana with its ARV program, the first and most advanced in sub-Saharan Africa. It identifies five features of the model and shows how they contributed to the ARV program. It also raises questions about ACHAP's role in scaling up and sustaining the program, as Botswana faces the challenges of treating growing numbers of HIV-infected people. PMID- 15757944 TI - Disease management's economic impact: unproven? PMID- 15757945 TI - Consolidation in health care. PMID- 15757946 TI - Moral hazard health spending. PMID- 15757951 TI - Toward functional proteomics of alveolar macrophages. AB - Alveolar macrophages (AM) belong to a phenotype of macrophages with distinct biological functions and important pathophysiological roles in lung health and disease. The molecular details determining AM differentiation from blood monocytes and AM roles in lung homeostasis are largely unknown. With the use of different technological platforms, advances in the field of proteomics have made it possible to search for differences in protein expression between AM and their precursor monocytes. Proteome features of each cell type provide new clues into understanding mononuclear phagocyte biology. In-depth analyses using subproteomics and subcellular proteomics offer additional information by providing greater protein resolution and detection sensitivity. With the use of proteomic techniques, large-scale mapping of phosphorylation differences between the cell types have become possible. Furthermore, two-dimensional gel proteomics can detect germline protein variants and evaluate the impact of protein polymorphisms on an individual's susceptibility to disease. Finally, surface enhanced laser desorption and ionization (SELDI) time-of-flight mass spectrometry offers an alternative method to recognizing differences in protein patterns between AM and monocytes or between AM under different pathological conditions. This review details the current status of this field and outlines future directions in functional proteomic analyses of AM and monocytes. Furthermore, this review presents viewpoints of integrating proteomics with translational topics in lung diseases to define the mechanisms of disease and to uncover new diagnostic and therapeutic targets. PMID- 15757952 TI - Pathogenesis of ventilator-induced lung injury: trials and tribulations. PMID- 15757953 TI - Pulmonary surfactant in allergic inflammation: new insights into the molecular mechanisms of surfactant function. PMID- 15757955 TI - Insulin vs. strict blood glucose control to achieve a survival benefit after AMI? PMID- 15757954 TI - Surfactant alterations in acute inflammatory lung injury from aspiration of acid and gastric particulates. AB - This study examines surfactant dysfunction in rats with inflammatory lung injury from intratracheal instillation of hydrochloric acid (ACID, pH 1.25), small nonacidified gastric particles (SNAP), or combined acid and small gastric particles (CASP). Rats given CASP had the most severe lung injury at 6, 24, and 48 h based on decreases in arterial oxygenation and increases in erythrocytes, total leukocytes, neutrophils, total protein, and albumin in bronchoalveolar lavage (BAL). The content of large surfactant aggregates in BAL was reduced in all forms of aspiration injury, but decreases were greatest in rats given CASP. Large aggregates from aspiration-injured rats also had decreased levels of phosphatidylcholine (PC) and increased levels of lyso-PC and total protein compared with saline controls (abnormalities for CASP were greater than for SNAP or ACID alone). The surface tension-lowering ability of large surfactant aggregates on a bubble surfactometer was impaired in rats with aspiration injury at 6, 24, and 48 h, with the largest activity reductions found in animals given CASP. There were strong statistical correlations between surfactant dysfunction (increased minimum surface tension and reduced large aggregate content) and the severity of lung injury based on arterial oxygenation and levels of albumin, protein, and erythrocytes in BAL (P < 0.0001). Surfactant dysfunction also correlated strongly with reduced lung volumes during inflation and deflation (P = 0.0004-0.005). These results indicate that surfactant abnormalities are functionally important in gastric aspiration lung injury and contribute significantly to the increased severity of injury found in CASP compared with ACID or SNAP alone. PMID- 15757956 TI - How should we evaluate an open artery in STEMI patients? PMID- 15757957 TI - Linkage between elevated PDGF-C expression and myocardial fibrogenesis in coxsackievirus B3-induced chronic myocarditis. PMID- 15757958 TI - Elevated expression of PDGF-C in coxsackievirus B3-induced chronic myocarditis. AB - AIMS: Coxsackievirus B3 (CVB3) is a frequent cause of human chronic myocarditis and subsequent fibrosis, leading to dilated cardiomyopathy. The molecular processes underlying the development of fibrosis are poorly understood. Enhanced levels of platelet-derived growth factors (PDGFs), especially PDGF-C, have recently been linked with the development of different forms of fibrosis. Therefore, the expression of PDGF was analysed in hearts of CVB3-infected major histocompatability complex class II knockout mice. The latter were recently established as mouse model mimicking the chronic inflammation and fibrosis characteristic for this disease. METHODS AND RESULTS: Expression of PDGF was analysed by reverse transcription-polymerase chain reaction, in situ hybridization, and immunohistochemistry. Hearts of C57BL/6 mice served as controls because infection of these animals leads to acute cardiac inflammation, but the hearts heal without signs of chronic inflammation. In uninfected hearts, basal expression of PDGF, notably PDGF-C, was detectable throughout the heart. The chronic inflammatory process was associated with elevated and sustained expression of all tested PDGF isoforms. Immunostaining and in situ hybridization analysis localized enhanced PDGF levels to areas with highest virus load and inflammatory infiltrations, adjacent to fibrotic areas. CONCLUSION: PDGF may participate in fibrosis development in CVB3-induced myocarditis. Therefore, PDGF signalling may be considered a target for therapeutic interference. PMID- 15757959 TI - Economic evaluation of nurse led intermediate care versus standard care for post acute medical patients: cost minimisation analysis of data from a randomised controlled trial. AB - OBJECTIVE: To undertake an economic evaluation of nurse led intermediate care compared with standard hospital care for post-acute medical patients. DESIGN: Cost minimisation analysis from an NHS perspective, comprising secondary care, primary care, and community care, using data from a pragmatic randomised controlled trial. SETTING: Nurse led unit and acute general medical wards in large, urban, UK teaching hospital. PARTICIPANTS: 238 patients. OUTCOME MEASURE: Costs to acute hospital trusts and to the NHS over six months. RESULTS: On an intention to treat basis, nurse led care was associated with higher costs during the initial admission period (nurse led care 7892 pounds sterling (14,970 dollars; 11,503 euros), standard care 4810 pounds sterling, difference 3082 pounds sterling (95% confidence interval 1161 pounds sterling to 5002 pounds sterling)). During the readmission period, costs were similar (nurse led care 1444 pounds sterling, standard care 1879 pounds sterling, difference -435 pounds sterling, -1406 pounds sterling to 536 pounds sterling). Total costs at six months were significantly higher (nurse led care 10,529 pounds sterling , standard care 7819 pounds sterling, difference 2710 pounds sterling, 518 pounds sterling to 4903 pounds sterling). Sensitivity analyses suggested that the trend for nurse led care to be more expensive was maintained even with substantial cost reductions, although differences were no longer significant. CONCLUSION: Acute hospitals may not be cost effective settings for nurse led intermediate care. Both inpatient and total costs were significantly higher for nurse led care than for standard care of post-acute medical patients, suggesting that this model of care should not be pursued unless clinical or organisational benefits justify the increased investment. PMID- 15757960 TI - Screening for abdominal aortic aneurysms: single centre randomised controlled trial. AB - OBJECTIVE: To determine whether screening Danish men aged 65 or more for abdominal aortic aneurysms reduces mortality. DESIGN: Single centre randomised controlled trial. SETTING: All five hospitals in Viborg County, Denmark. PARTICIPANTS: All 12,639 men born during 1921-33 and living in Viborg County. In 1994 we included men born 1921-9 (64-73 years). We also included men who became 65 during 1995-8. INTERVENTIONS: Men were randomised to the intervention group (screening by abdominal ultrasonography) or control group. Participants with an abdominal aortic aneurysm > 5 cm were referred for surgical evaluation, and those with smaller aneurysms were offered annual scans. OUTCOME MEASURES: Specific mortality due to abdominal aortic aneurysm, overall mortality, and number of planned and emergency operations for abdominal aortic aneurysms. RESULTS: 4860 of 6333 men were screened (attendance rate 76.6%). 191 (4.0% of those screened) had abdominal aortic aneurysms. The mean follow-up time was 52 months. The screened group underwent 75% (95% confidence interval 51% to 91%) fewer emergency operations than the control group. Deaths due to abdominal aortic aneurysms occurred in nine patients in the screened group and 27 in the control group. The number needed to screen to save one life was 352. Specific mortality was significantly reduced by 67% (29% to 84%). Mortality due to non-abdominal aortic aneurysms was non-significantly reduced by 8%. The benefits of screening may increase with time. CONCLUSION: Mass screening for abdominal aortic aneurysms in Danish men aged 65 or more reduces mortality. PMID- 15757961 TI - Disease- and cell-type-specific transcriptional targeting of vectors for osteoarthritis gene therapy: further development of a clinical canine model. AB - OBJECTIVES: The potential for undesirable systemic effects related to constitutive expression of certain therapeutic transgenes may be limited through the development of transcriptionally targeted disease- and cell-type-specific vectors. The objective of this study was to analyse the canine matrix metalloproteinase-9 (MMP-9) promoter and deletion constructs for its ability to drive expression in response to pro-inflammatory cytokines (interleukin-1beta and tumour necrosis factor-alpha). METHODS: Initial analysis of MMP-9 deletion constructs was made using a luciferase reporter system. The promoter was subsequently engineered to incorporate multiple NF-kappaB sites. In parallel experiments we used the mouse collagen type XI promoter to study cell-type specific promoter activity in chondrocyte-specific cells (SW1353) and undifferentiated chondroprogenitor cells (ATDC5). RESULTS: Incorporation of multiple NF-kappaB sites into the MMP-9 promoter enhanced activity while maintaining disease specificity. Further, manipulation of the mouse collagen type XI (mColXI) promoter by the incorporation of SOX9 enhancer sites downstream of a reporter gene, increased gene activity while maintaining cell type specificity. CONCLUSIONS: Manipulation of promoter and enhancer regions can improve transcriptionally targeted genes. A combination of these systems, in the context of the canine model, has the potential to improve the safety of osteoarthritis gene therapy vectors. PMID- 15757962 TI - Measuring disease activity in ankylosing spondylitis: patient and physician have different perspectives. AB - OBJECTIVE: There is no "gold standard" to assess disease activity in patients with ankylosing spondylitis (AS). It is known that patients and physicians have different opinions about disease activity. The objective was therefore to investigate on which criteria patients with AS and physicians base their judgement on disease activity. METHODS: A cohort of 203 AS out-patients fulfilling the modified New York criteria included in the ongoing long-term follow-up was analysed. The Assessment in Ankylosing Spondylitis (ASAS) International Working Group has established different domains relevant for outcome in AS. Each domain includes a number of instruments for making assessments, and all these instruments are included in the Outcome in Ankylosing Spondylitis International Study and were made every 6 months for 2 yr. Disease activity from the patient perspective as well as from the physician perspective was analysed using the patient's or the physician's global assessment of disease activity [visual analogue scale (VAS): 0 (best)-10 (worst)] by dichotomizing into "high disease activity" (VAS > or = 6.0) and "low disease activity" (VAS < or = 4.0). Data reduction by principal components analysis (PCA) was performed to distinguish factors capturing correlated instruments. Discriminant analysis with the factor loadings was performed to discriminate between a low and a high disease activity state from both the patient's and the physician's perspective. Multiple regression analysis on the discriminant scores was performed to prioritize the instruments. RESULTS: PCA revealed four factors: spinal mobility, physician assessments, patient assessments and laboratory assessments (Cronbach's alpha 0.52-0.80; explained variance 61%). Discriminant function analysis showed that the factor "patient assessments" was most important (pooled correlation 0.85) in discriminating between a low and a high disease activity state as defined by the patient. The other three factors contributed marginally (pooled correlation <0.30). In contrast, the factors "physician's assessments" (pooled correlation 0.62), "spinal mobility" (pooled correlation 0.52) and "laboratory assessments" (pooled correlation 0.48) contributed most to the physician's perspective. The factor "patient assessments" did not contribute at all (pooled correlation 0.05). Multivariate analysis on the discriminant scores showed that the instruments "pain spine", "BASFI", "pain joints" and "BASDAI fatigue" explained more than 90% of variance in the case of the patient perspective. The instruments "cervical rotation", "swollen joint count", "CRP" and "intermalleolar distance" explained more than 90% of variance in case of physician perspective. CONCLUSION: AS patients rate disease activity on the basis of complaints while physicians rate disease activity on the basis of instruments related to disease severity and inflammation. PMID- 15757963 TI - Recurrence risk for psoriasis and psoriatic arthritis within sibships. AB - OBJECTIVE: To quantify the frequency of siblings of patients with psoriatic arthritis (PsA) having psoriasis and/or inflammatory arthritis. To describe the similarity or otherwise of patterns of arthritis in siblings. METHODS: Available and consenting index cases with PsA and one or more siblings living locally were assessed. Mean sibling concordance rates and Weinberg's segregation analysis were calculated. Heritability was also estimated. To assess whether the same type of arthritis occurred within the same sibship, the dually affected sibships were then classified for type of arthritis according to methods suggested by Moll, Helliwell, Veale and McGonagle. RESULTS: Eighty index cases and 112 siblings were assessed. The median age of index cases was 49 yr (range 24-80 yr) and for siblings 46 yr (range 18-79 yr). The concordance rate for all types of PsA was 14% (9% if enthesitis is excluded) and for psoriasis 21%. There was no difference in the two methods used to calculate concordance rates. Sixteen dually affected sib pairs were found. Four of the 16 sibships (25%) had the same pattern of joint involvement (Moll and Wright classification). The most frequent pattern seen was joint involvement identical to rheumatoid arthritis (3/5). The most common symptom in affected siblings was enthesitis (approximately 5%). When the dually affected sibships were analysed using the other classifications, the simpler the classification the greater the concordance for joint pattern. CONCLUSION: The concordance for psoriasis is greater than for PsA, but the concordance rate for PsA was similar to that in HLA identical siblings with rheumatoid arthritis. There was discordance in pattern of arthritis for most sib pairs. There is no support for the use of more complex classifications of PsA. PMID- 15757964 TI - Leflunomide-induced subacute cutaneous lupus erythematosus. PMID- 15757966 TI - Down-regulation of activating Fcgamma receptors on monocytes of patients with rheumatoid arthritis upon methotrexate treatment. AB - OBJECTIVE: To determine the effect of methotrexate (MTX) on expression levels of activating receptors for IgG (FcgammaRs) on monocytes of rheumatoid arthritis (RA) patients in relation to changes in disease activity. METHODS: The effect of MTX on FcgammaRs on monocytes of RA patients was evaluated ex vivo as well as in vitro. Recently diagnosed, disease-modifying antirheumatic drug (DMARD)-naive RA patients were treated with low-dose MTX. At baseline and 16 weeks after the start of MTX treatment, changes in FcgammaR expression levels on peripheral blood monocytes were evaluated by fluorescence-activated cell sorting analysis and were correlated to changes in disease parameters. To study the direct effects of MTX on monocytes, these cells were isolated from peripheral blood monocytes of healthy controls and cultured with MTX. Other monocyte surface molecules (CD40, CD80, CD86, MHC class II) were also determined to test the specificity of the effect on FcgammaR expression levels. RESULTS: Eleven out of 15 patients improved clinically (mean disease activity score before 6.2 +/- 0.8 vs 4.3 +/- 1.7 after). Sixteen weeks after the start of MTX therapy, the expression levels of FcgammaRI and IIa on monocytes were significantly decreased, whereas the decreases in FcgammaRIIIa expression levels on monocytes were less marked. The percentage decrease in FcgammaRI expression correlated with the percentage decrease in CRP and well-being. In vitro MTX selectively decreased FcgammaRI and FcgammaRIIa expression levels of isolated monocytes, in contrast to other surface molecules. CONCLUSION: The disease-modifying effect of MTX in the treatment of RA is accompanied by down-regulation of activating FcgammaRI and IIa on monocytes, which could be a direct effect of MTX on monocytes. This down-regulation represents a new mode of action of MTX which should be considered in RA patients, especially during conditions that could give rise to monocyte activation by IgG containing immune complexes, e.g. during antibody-based therapy of RA. PMID- 15757965 TI - Persistent clinical response to the anti-TNF-alpha antibody infliximab in patients with ankylosing spondylitis over 3 years. AB - OBJECTIVE: Infliximab, a monoclonal antibody against tumour necrosis factor alpha (TNF-alpha), is approved in Europe for the treatment of patients with active ankylosing spondylitis (AS) who have responded inadequately to conventional therapy. This report provides analyses from a 3-yr extension study, as a follow up to both the 1- and 2-yr open label extensions of the original 3-month randomized controlled trial of infliximab in patients with AS. METHODS: Of the 49 patients with AS who completed the second year of the study, 46 continued treatment with infliximab 5 mg/kg every 6 weeks up to week 156. The Bath AS Disease Activity Index (BASDAI), the Bath AS Functional Index, the Bath AS Metrology Index, patient's and physician's global assessments, quality of life (Short Form-36), C-reactive protein (CRP) and erythrocyte sedimentation rate were assessed throughout the study period. RESULTS: The improvement of signs and symptoms observed in the majority of the patients during the first and second year was sustained throughout the third year of the study. Forty-three patients (62% of the 69 patients enrolled at baseline and 93% of the patients who started the third year) completed week 156. In the intention-to-treat analysis, an ASAS '5 out of 6' and ASAS 40% response was seen by 46% and 50% of the patients, respectively. The scores for other efficacy assessments were similar to the values observed at weeks 54 and 102. Median CRP levels remained low (1.5 mg/l at week 156). There were no relevant side-effects and no discontinuation because of drug-related adverse events during the third year of the study. CONCLUSIONS: Patients with AS receiving infliximab for 3 yr showed a durable clinical response without loss of efficacy. Long-term infliximab treatment was well tolerated by patients in this study. PMID- 15757967 TI - Preclinical vascular disease in systemic lupus erythematosus and primary antiphospholipid syndrome. AB - OBJECTIVE: To determine the prevalence of preclinical vascular disease and associated risk factors in patients with systemic lupus erythematosus (SLE) or primary antiphospholipid syndrome (APS). METHODS: We consecutively studied 70 SLE patients and 25 primary APS patients without clinical coronary artery disease. The control group included 40 healthy women. Carotid ultrasound was performed and the intima-media wall thickness (IMT) and presence of plaque was investigated in all patients and controls. Traditional vascular risk factors and SLE-disease and treatment related factors were also analysed. RESULTS: SLE patients had a higher prevalence of traditional atherosclerosis risk factors: hypertension (P<0.005) and dyslipidaemia (P<0.05) and higher levels of total cholesterol (P = 0.03), triglycerides (P = 0.004) and apolipoprotein B (P = 0.04). The prevalence of carotid plaque was higher and appeared earlier in SLE patients than in the primary APS patients or controls (P<0.001). The IMT was similar in the three groups. SLE patients with secondary APS had a higher prevalence of carotid plaque than patients with primary APS (37.5% vs 8%, P = 0.03). The presence of plaque in SLE patients was associated with a higher SLICC score (2.40 +/- 1.78 vs 1.02 +/- 1.18, P = 0.002), higher ECLAM score (3.10 +/- 2.32 vs 1.84 +/- 1.59, P = 0.02) and older age (47.3 +/- 8.44 vs 37.38 +/- 11.28, P = 0.003) at the time of carotid ultrasound study. CONCLUSION: Plaque prevalence in patients with primary APS is similar to that of controls and inferior to that of SLE patients with secondary APS. SLE patients have a high prevalence of early carotid atherosclerosis that is associated with cumulative disease damage and disease activity. PMID- 15757969 TI - Meeting the challenges of research governance. PMID- 15757968 TI - The systemic lupus erythematosus tri-nation study: longitudinal changes in physical and mental well-being. AB - OBJECTIVE: We have shown that SLE patients in Canada and the UK incurred 20% and 13% lower health costs than those in the US, respectively, but did not experience worse outcomes as expressed by the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index. We now compare change in quality of life in these patients. PATIENTS AND METHODS: Seven hundred and fifteen SLE patients (Canada 231, US 269, UK 215) completed the SF-36 annually over four years. The annual change in the SF-36 Physical and Mental Component Summary (PCS and MCS) scores over the course of the study were summarized by estimating a linear trend for each individual patient using hierarchical modelling. Cross-country comparison of the slopes in the PCS and MCS scores was then performed using simultaneous regressions. RESULTS: The estimated mean annual changes (95% credible interval [CrI]) in the PCS scores in Canada, the US, and the UK were 0.18 (-0.07, 0.43), -0.05 (-0.27, 0.17), and 0.03 (-0.20, 0.27), respectively; the mean annual changes in the MCS scores were 0.15 (-0.04, 0.34), 0.23 (0.09, 0.37), and 0.08 (-0.10, 0.27), respectively. Regression results showed that the mean annual changes in PCS and MCS scores did not substantially differ across countries. CONCLUSION: Quality of life remained stable across countries. Despite Canadian and British patients incurring lower health costs, on average, patients experienced similar changes in physical and mental well-being. PMID- 15757970 TI - Validity and reliability of an Italian version of the revised Leeds disability questionnaire for patients with ankylosing spondylitis. AB - OBJECTIVE: The purpose of the present study was to produce an Italian version of the Revised Leeds Disability Questionnaire (LDQ) in a group of patients with ankylosing spondylitis, and to examine the psychometric properties of this version, evaluating its internal consistency, external validity and reliability. METHODS: The LDQ was administered to 60 Caucasian patients affected by ankylosing spondylitis (50 males, 10 females, mean age 46.1 +/- 14.2 yr, range 22-74, median disease duration 4.5 yr, range 1-24) together with the Italian version of the Stanford Health Assessment Questionnaire (HAQ), and anthropometric measurements. Thirty patients completed the questionnaire after a 10-day interval. Internal consistency was evaluated with Cronbach's alpha coefficient of reliability. Construct validity of the LDQ was evaluated using the correlation between the HAQ and anthropometric measurements. Test-retest reliability was assessed with the intraclass correlation coefficient. RESULTS: All patients completed the validation study. The questionnaire was internally consistent (alpha=0.90). A significant correlation was recorded between the LDQ and the HAQ score (rho=0.841, P<0.01) and the anthropometric measurements. Test-retest reliability showed a good correlation coefficient (intraclass correlation=0.97). CONCLUSION: The Italian LDQ is a valid and reliable instrument for detecting and measuring functional disability in patients with ankylosing spondylitis. Our results confirm the utility of this questionnaire as a valid and feasible functional measure for patients with ankylosing spondylitis. PMID- 15757971 TI - Successful treatment of steroid-resistant Weber-Christian disease with biliary ductopenia using cyclosporin A. PMID- 15757972 TI - A cell-based screen for modulators of ataxin-1 phosphorylation. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal dominant neurodegenerative disorder caused by the expansion of a glutamine repeat within the SCA1-encoded protein ataxin-1. We have previously shown that serine 776 (S776) of both wild type and mutant ataxin-1 is phosphorylated in vivo and in vitro. Moreover, preventing phosphorylation of this residue by replacing it with alanine resulted in a mutant protein, which was not pathogenic in spite of its nuclear localization. To further investigate pathways leading to S776 phosphorylation of ataxin-1, we developed a cell-culture based assay to screen for modulators of S776 phosphorylation. In this assay, ataxin-1 expression was monitored by enhanced green fluorescent protein (EGFP) fluorescence in cell lines stably expressing EGFP-ataxin-1 fusion protein. The phospho-S776 ataxin-1 specific antibody (PN1168) was used to assess ataxin-1 S776 phosphorylation. A library of 84 known kinase and phosphatase inhibitors was screened. Analysis of the list of drugs that modified S776 phosphorylation places many of the inhibited kinases into known cell signaling pathways. A pathway associated with calcium signaling resulted in phosphorylation of both wild-type and mutant ataxin-1. Interestingly, inhibitors of the PI3K/Akt pathway predominantly diminished mutant ataxin-1 phosphorylation. These results provide new molecular tools to aid in elucidating the biological role of ataxin-1 phosphorylation and perhaps provide potential leads toward the development of a therapy for SCA1. PMID- 15757973 TI - Abnormal Ca(2+) release and catecholamine-induced arrhythmias in mitochondrial cardiomyopathy. AB - Mitochondrial dysfunction is implicated in numerous cardiac disorders. It has been assumed that the functional defects are directly related to a decreased rate of mitochondrial ATP production, but recent studies have challenged this idea. Here, we used mice with tissue-specific knockout of mitochondrial transcription factor A (Tfam) that leads to progressive cardiomyopathy. The role of changes in the excitation-contraction (E-C) coupling in cardiomyocytes of these mice was studied by measuring the free cytosolic Ca(2+) concentration and by analyzing the expression of genes encoding E-C coupling proteins. Action potential-mediated Ca(2+) transients, measured with the fluorescent indicator fluo-3 in isolated cardiomyocytes, were smaller and faster in Tfam knockout cardiomyocytes when compared with controls. The total sarcoplasmic reticulum (SR) Ca(2+) content was decreased in Tfam knockout cells. The gene for the SR Ca(2+) binding protein calsequestrin-2 (CASQ2), as well as other genes encoding proteins involved in SR Ca(2+) handling, showed decreased expression in Tfam knockout hearts. Decreased CASQ2 levels have been linked to severe arrhythmias triggered by beta-adrenergic stimulation. In line with this, application of the beta-adrenergic agonist isoproterenol resulted in frequent doublet Ca(2+) transients in Tfam knockout cardiomyocytes. In conclusion, our results show that mitochondrial dysfunction in the heart induces specific down-regulation of the expression of genes encoding proteins involved in E-C coupling. These changes predispose to cardiac arrhythmias and terminal heart failure and are thus important in the pathogenesis of mitochondrial cardiomyopathy. PMID- 15757974 TI - Transdifferentiation of the retinal pigment epithelia to the neural retina by transfer of the Pax6 transcriptional factor. AB - The Pax6 gene plays an important role in eye morphogenesis throughout the animal kingdom. The Pax6 gene and its homologue could form ectopic eyes by targeted expression in Drosophila and Xenopus. Thus, this gene is a master gene for the eye morphogenesis at least in these animals. In the early development of the vertebrate eye, Pax6 is required for the instruction of multipotential progenitor cells of the neural retina (NR). Primitive retinal pigment epithelial (RPE) cells are able to switch their phenotype and differentiate into NR under exogenous intervention, including treatment with fibroblast growth factors (FGFs), and surgical removal of endogenous NR. However, the molecular basis of phenotypic switching is still controversial. Here, we show that Pax6 alone is sufficient to induce transdifferentiation of ectopic NR from RPE cells without addition of FGFs or surgical manipulation. Pax6-mediated transdifferentiation can be induced even at later stages of development. Both in vivo and in vitro studies show that the Pax6 lies downstream of FGF signaling, highlighting the central roles of Pax6 in NR transdifferentiation. Our results provide an evidence of retinogenic potential of nearly mature RPE and a cue for new therapeutic approaches to regenerate functional NR in patients with a visual loss. PMID- 15757976 TI - The pathogenic agent in Drosophila models of 'polyglutamine' diseases. AB - A substantial body of evidence supports the identity of polyglutamine as the pathogenic agent in a variety of human neurodegenerative disorders where the mutation is an expanded CAG repeat. However, in apparent contradiction to this, there are several human neurodegenerative diseases (some of which are clinically indistinguishable from the 'polyglutamine' diseases) that are due to expanded repeats that cannot encode polyglutamine. As polyglutamine cannot be the pathogenic agent in these diseases, either the different disorders have distinct pathogenic pathways or some other common agent is toxic in all of the expanded repeat diseases. Recently, evidence has been presented in support of RNA as the pathogenic agent in Fragile X-associated tremor/ataxia syndrome (FXTAS), caused by expanded CGG repeats at the FRAXA locus. A Drosophila model of FXTAS, in which 90 copies of the CGG repeat are expressed in an untranslated region of RNA, exhibits both neurodegeneration and similar molecular pathology to the 'polyglutamine' diseases. We have, therefore, explored the identity of the pathogenic agent, and specifically the role of RNA, in a Drosophila model of the polyglutamine diseases by the expression of various repeat constructs. These include expanded CAA and CAG repeats and an untranslated CAG repeat. Our data support the identity of polyglutamine as the pathogenic agent in the Drosophila models of expanded CAG repeat neurodegenerative diseases. PMID- 15757975 TI - MeCP2 deficiency in Rett syndrome causes epigenetic aberrations at the PWS/AS imprinting center that affects UBE3A expression. AB - Rett syndrome (RS) is a severe and progressive neurodevelopmental disorder caused by heterozygous mutations in the X-linked methyl CpG binding protein 2 (MeCP2) gene. MeCP2 is a nuclear protein that binds specifically to methylated DNA and functions as a general transcription repressor in the context of chromatin remodeling complexes. RS shares clinical features with those of Angelman syndrome (AS), an imprinting neurodevelopmental disorder. In AS patients, the maternally expressed copy of UBE3A that codes for the ubiquitin protein ligase 3A (E6-AP) is repressed. The similar phenotype of these two syndromes led us to hypothesize that part of the RS phenotype is due to MeCP2-associated silencing of UBE3A. Indeed, UBE3A mRNA and protein are shown here to be significantly reduced in human and mouse MECP2 deficient brains. This reduced UBE3A level was associated with biallelic production of the UBE3A antisense RNA. In addition, MeCP2 deficiency resulted in elevated histone H3 acetylation and H3(K4) methylation and reduced H3(K9) methylation at the PWS/AS imprinting center, with no effect on DNA methylation or SNRPN expression. We conclude, therefore, that MeCP2 deficiency causes epigenetic aberrations at the PWS imprinting center. These changes in histone modifications result in loss of imprinting of the UBE3A antisense gene in the brain, increase in UBE3A antisense RNA level and, consequently reduction in UBE3A production. PMID- 15757977 TI - Muscle-specific BCL2 expression ameliorates muscle disease in laminin {alpha}2 deficient, but not in dystrophin-deficient, mice. AB - To examine the role of apoptosis in neuromuscular disease progression, we have determined whether pathogenesis in dystrophin-deficient (mdx) and laminin alpha2 deficient (Lama2-null) mice is ameliorated by overexpression of the anti apoptosis protein BCL2 in diseased muscles. The mdx mice are a model for the human disease, Duchenne muscular dystrophy (DMD), and the Lama2-null mice are a model for human congenital muscular dystrophy type 1A (MDC1A). For these studies, we generated transgenic mice that overexpressed human BCL2 under control of muscle-specific MyoD or MRF4 promoter fragments. We then used cross-breeding to introduce the transgenes into diseased mdx or Lama2-null mice. In mdx mice, we found that overexpression of BCL2 failed to produce any significant differences in muscle pathology. In contrast, in the Lama2-null mice, we found that muscle specific expression of BCL2 led to a several-fold increase in lifespan and an increased growth rate. Thus, BCL2-mediated apoptosis appears to play a significant role in pathogenesis of laminin alpha2 deficiency, but not of dystrophin deficiency, suggesting that therapies designed to ameliorate disease by inhibition of apoptosis are more likely to succeed in MDC1A than in DMD. PMID- 15757978 TI - Mortality experience of male workers at a UK tin smelter. AB - BACKGROUND: Between 1937 and 1991, Capper Pass and Sons Limited operated a tin smelter complex in North Humberside, UK, at which employees were potentially exposed to a number of substances, including lead, arsenic, cadmium and natural series radionuclides. Decommissioning and site clearance continued until 1995. Between 1967 and 1995 the company was a subsidiary of Rio Tinto plc. AIMS: The aim was to identify any significant excess, or deficits, in mortality among former employees that might be attributable to factors associated with occupation. METHODS: We defined a cohort of 1462 males who had been employed for at least 12 months between 1/11/1967 and 28/7/1995, followed-up through to 31/12/2001. The mortality of the cohort was compared against that expected for both national and regional populations. RESULTS: Mortality from all causes and all cancers did not differ from that expected. Mortality from ischaemic heart disease showed a deficit and mortality from lung cancer showed a statistically significant excess. Mortality from smoking related diseases other than lung cancer showed a non-significant deficit. CONCLUSIONS: The pattern of lung cancer mortality is consistent with the hypothesis that the risk of lung cancer has been enhanced by occupational exposure to one or more carcinogens, the effect of which diminishes with time since exposure. The deficit in ischaemic heart disease may be attributed to a protective effect from manual labour. The results provide no evidence for attribution of other excess or deficits in mortality to factors associated with employment. PMID- 15757979 TI - Occupational medicine and the general practitioner. PMID- 15757981 TI - Latent period for symptomatic sensitization in bakeries. AB - AIM: To describe the latent period for symptomatic sensitization in bread bakery workers. METHODS: Data on the latent period for symptomatic sensitization, resulting in either asthma or rhinitis, were obtained from an in-house health surveillance programme in a single large organization. RESULTS: Over a period of 10 years, 90 employees were identified with symptoms attributable to sensitization. The mean latent period was 7.3 years, with three employees describing the onset of symptoms in their first year of exposure. CONCLUSIONS: The long latent period, typically more than 1 year, suggests that health surveillance on appointment and then at annual intervals thereafter should be adequate to identify affected individuals in a bakery environment. PMID- 15757980 TI - Mesothelioma mortality in Great Britain from 1968 to 2001. AB - BACKGROUND: The British mesothelioma register contains all deaths from 1968 to 2001 where mesothelioma was mentioned on the death certificate. AIMS: To present summary statistics of the British mesothelioma epidemic including summaries by occupation and geographical area. METHODS: Standardized mortality ratios (SMRs) were calculated for local authorities, unitary authorities and counties. Temporal trends in SMRs were also examined. Proportional mortality ratios (PMRs) were calculated using the Southampton (based on the 1980 standard occupational classification) coding scheme. Temporal trends in PMRs were also examined. RESULTS: The annual number of mesothelioma deaths has increased from 153 in 1968 to 1848 in 2001. Current deaths in males account for about 85% of the cases. The areas of West Dunbartonshire (SMR 637), Barrow-in-Furness (593), Plymouth (396) and Portsmouth (388) have the highest SMRs over the period 1981-2000. The occupations with the highest PMRs are metal plate workers (PMR 503), vehicle body builders (526), plumbers and gas fitters (413) and carpenters (388). CONCLUSIONS: These data reinforce earlier findings that geographical areas and occupations associated with high exposure to asbestos in the past continue to drive the mesothelioma epidemic in Great Britain. However, the trends over time suggest a change in the balance of risk away from traditional asbestos exposure industries to industries where one could describe the exposure as secondary, such as plumbers and gas fitters, carpenters, and electricians. PMID- 15757982 TI - The use of NHS accident and emergency services by commercial sea fishermen in the North East of Scotland. AB - BACKGROUND: Commercial sea fishing is known to be a hazardous occupation, but little quantitative research has been conducted in the UK. OBJECTIVE: To describe the demography of commercial fishermen accessing NHS accident and emergency (A&E) services and profile the reasons for attendance. METHODS: Eight NHS A&E departments in North East Scotland participated in the audit. A structured data collection form was completed by the attending nurse or doctor during a 6 month period (March-August). All commercial sea fishermen attending during this time were included whether or not the reason for attendance was work-related. RESULTS: There were 164 cases of which 133 (81%) presented with injuries and 19 (12%) with illnesses (12 unknown). Twenty-seven (16%) cases had required emergency evacuation from commercial vessels. The most frequent category of injury was 'hand', 'wrist' or 'finger' followed by 'head', 'face' and 'throat'. CONCLUSION: Commercial sea fishing is a dangerous occupation with many injuries requiring NHS A&E treatment. The relatively high level of injuries compared to illness indicates that there are still safety issues to be addressed and current risk assessment requirements need to be reviewed. PMID- 15757983 TI - Minor injuries, cognitive failures and accidents at work: incidence and associated features. AB - BACKGROUND: There is a dearth of information examining minor injuries and cognitive failures in the workplace. AIMS: This study aims to describe the incidence of, and characteristics associated with, minor injuries, cognitive failures and accidents in the workplace. METHODS: The study was a community based postal questionnaire survey of people selected at random from the Electoral Registers of Cardiff and Merthyr Tydfil; 7980 questionnaires were returned. RESULTS: Frequent/very frequent minor injuries were reported by 9.8% of the workers and frequent/very frequent cognitive failures (e.g. problems of memory, attention or action) by 10.5%. Work accidents requiring treatment were reported by 5.6% of the workers. Minor injuries, cognitive failures and accidents were associated with a number of demographic and health factors. In addition, minor injuries, cognitive failures and accidents were found to be related to each other. CONCLUSIONS: A large number of workers experience accidents and minor injuries at work. Minor injuries are not recorded by official sources but could have implications in productivity and worker health. Factors identified in this study, as associated with injuries or accidents, could be addressed by specific workplace policies or by raising individual awareness of the risks. PMID- 15757984 TI - Skin disease in paper mill workers. AB - BACKGROUND: Paper mill workers have frequent and prolonged exposure to skin irritants and allergens and may have a higher risk of developing occupational dermatitis. Aims The aim of this study was to determine the extent of skin problems in a paper mill and how much was attributable to contact with allergens. METHODS: A cross-sectional study was carried out among 80 paper mill workers having daily exposure to skin irritants and allergens. They all completed a questionnaire, underwent a standard interview and physical examination. Workers whose history indicated possible contact allergy were patch tested and prick tested. RESULTS: Workers reported a high exposure to skin irritants, especially when carrying out tasks that caused the hands and feet to become wet from perspiration and having contact with process water. Atopic dermatitis was seen in 3% of the workers. Contact dermatitis was seen in 26% of the workers and 36% were diagnosed with mycosis of the feet. All cases of contact dermatitis and mycosis could be attributed to occupational exposure to skin irritants. No cases of relevant contact allergy were seen. CONCLUSION: Occupational dermatitis in paper mills is primarily related to the exposure to skin irritants. Occupational physicians should be aware of the risk of occupational dermatitis in paper mill workers. PMID- 15757986 TI - Musculoskeletal disorders and visual strain in intensive data processing workers. AB - BACKGROUND: This study was conducted for a UK Trade Union in response to increasing numbers of health complaints among intensive computer workers in a data processing environment. AIM: To estimate prevalence of musculoskeletal pain/discomfort and visual strain symptoms among data processing workers and to explore associations with work factors. METHODS: A self-report questionnaire was distributed to data processors (n = 175) and a control group (n = 129) in the same organization. RESULTS: Eighty-six per cent of data processors reported musculoskeletal pain/discomfort in the previous year, with the highest prevalence rate found for the neck (58%). The 12 month reported prevalence for visual strain symptoms was 47%, tired eyes (41%) was the most commonly reported symptom. In the last week, 56% reported pain/discomfort and 25% visual strain symptoms. Data processors reporting pain in the previous year were more likely to be dissatisfied with their jobs (P < 0.01), to report a lack of choice in deciding what they did at work (P < 0.02), to have insufficient time to complete work (P < 0.05) or to receive help from others when time was limited (P < 0.03). CONCLUSION: The prevalence of self-reported musculoskeletal pain/discomfort and visual strain symptoms was high among data processors. A systematic approach to risk reduction addressing organizational, psychosocial and physical work factors is required. This study has implications for reorganization of this and other intensive computer work, given the increasing requirement for intensive computing work in many industrial sectors. More research is needed to investigate interactions between job demands and support systems for these workers. PMID- 15757985 TI - Stress in the Norwegian police service. AB - BACKGROUND: High levels of stress have been attributed to the conditions of police work, but there is little empirical evidence for this. AIMS: To develop a new instrument to measure job stress in the police; to assess the most severe and frequent police stressors; to compare levels of stress according to the demographic and organizational factors; and to study stress in relation to personality traits, work locus of control and coping strategies. METHODS: A comprehensive nationwide questionnaire survey of 3272 Norwegian police at all hierarchical levels, including the Norwegian Police Stress Survey (NPSS), the Job Stress Survey, the Basic Character Inventory, the Work Locus of Control Scale, and the Coping Strategies Scale. RESULTS: Work injuries were appraised as the most stressful but least frequent stressor and job pressure was reported the least severe but most frequent stressor. Females experienced job stressors less frequently, but appraised them as more severe than men did. Older police officers reported more job pressure severity and fewer work injuries. The police in districts where peer support was planned but not implemented, and who worked in districts with more than 50,000 inhabitants, perceived the lack of support more severely than others. The correlations between stress and personality traits, work locus of control, and coping were moderate. CONCLUSIONS: The NPSS captures police-specific stressors that are not adequately measured by global stress instruments. The study of stress in police work should preferably involve a nationwide use of police-specific stress instruments. PMID- 15757987 TI - Direct health costs of occupationally acquired malaria in a military population in Europe. AB - BACKGROUND: We reviewed imported malaria cases in British troops stationed in Germany, to quantify the occupational health burden of malaria and audit the quality of local care. METHODS: We used a capture-recapture technique to ascertain imported malaria infections acquired by our military population during a 2-year period of surveillance (2001 and 2002). We retrieved and analysed all hospital discharge summaries. We calculated direct treatment costs from bills raised by the admitting German hospitals. RESULTS: Out of 24 600 British troops stationed in Germany, approximately 800 were occupationally exposed to malaria during 2001 and 800 during 2002. The mean duration of exposure was 1.5 months. All the occupational exposures in this period occurred during routine training exercises in the tropics and subtropics, or during short-term peacekeeping missions in endemic areas. There were three imported malaria cases in British soldiers during 2001 and 12 during 2002 (crude acquisition rates 3.75 and 15/1000). Two soldiers (one with Plasmodium vivax, the other with Plasmodium falciparum infection) required intensive hospital therapy. The median length of hospital inpatient stay was 7 days (for P. vivax infection) and 8.5 days (for P. falciparum). The direct treatment costs of the hospitalizations totalled 27,760. CONCLUSIONS: Comparing crude acquisition rates, non-immune soldiers carrying out peacetime activities in endemic areas may be at around 2-10 times greater risk of acquiring malaria than civilian travellers to the tropics. All military branches are at some risk, and the Royal Engineers may be at an especially high risk of occupationally acquired malaria. Better prevention is mandatory. PMID- 15757988 TI - Detailed calculation of occupational health service costs through activity-based costing: the cost of risk-assessment projects. PMID- 15757989 TI - Hepatitis B transmission through blood and body fluids exposure of school personnel. AB - BACKGROUND: Hepatitis B transmission from students to members of staff has been documented in schools, particularly nurseries and day care centres. AIMS: To investigate the frequency of exposure to blood and other body fluids within day schools and to document practices adopted by school personnel to avoid direct contact and decontaminate the environment. METHODS: Questionnaire survey among 21 public day schools in Malta. RESULTS: Episodes of significant blood exposure were rare, occurring at frequencies of 0.071 [95% confidence interval (CI): 0-0.148] incidents per thousand student days. Contact with larger volumes of other body fluids, namely urine and vomitus, was more likely: 0.12 (95% CI: 0.008-0.383) and 0.088 (95% CI: 0.048-0.128) episodes per 1000 student days, respectively. School personnel generally used correct personal protective equipment, particularly gloves, in cases of contact with blood and body fluids. Environmental disinfection methods varied considerably with only 38% of schools (95% CI: 21 59%) using recommended hypochlorite preparations. CONCLUSIONS: Exposure to quantities of blood sufficient to result in HBV transmission in day schools is rare. Emphasis should be placed on risk assessment at individual school level, concentrating on correct management of body fluid exposures through effective staff education. PMID- 15757990 TI - Correlates of physical activity among middle-aged Finnish male police officers. AB - BACKGROUND: Few studies exist on adherence to exercise within specific occupational groups. AIMS: This study aimed at identifying factors explaining leisure-time physical activity among middle-aged Finnish police officers. METHODS: Middle-aged police officers (n = 103) participated in the study in 1981 and 1996. Frequency of and adherence to leisure-time physical activity and fitness were assessed. RESULTS: The factor 'enjoyment' was the most powerful determinant for both physical activity and fitness, but all the factors studied accounted for only 10% of the variability. Leisure-time physical activity in 1981 correlated significantly with leisure-time physical activity in 1996. CONCLUSIONS: The physical activity of the middle-aged police officers can be in part predicted from their physical activity in early adulthood. Enjoyment related to physical activity seems to be important with respect to enhancing adherence. Perhaps, being fit increases the feeling of enjoyment during exercise. PMID- 15757991 TI - A screening questionnaire for HAVS? AB - AIM: To define the diagnostic power of simple questions most applicable for a hand-arm vibration syndrome (HAVS) assessment screening questionnaire. METHOD: Using a binary logistic regression we analysed 365 physician led HAVS health surveillance assessments to identify which questions could form the basis of a screening questionnaire. RESULTS: Four sensorineural related questions regarding tingling and numbness in response to the cold and after using vibrating tools, and two vascular-related questions focusing on the patient's fingers going white on exposure to cold and numbness during an attack of whiteness were identified. CONCLUSIONS: Questions of high sensitivity for screening subjects for the vascular and neurosensory components of HAVS were identified, which can be used to identify those requiring further clinical investigation and functional testing. PMID- 15757992 TI - Anosmia in association with occupational use of a waterproof coating chemical. AB - A case of acute permanent anosmia is described in a renovation worker during exposure to a waterproof coating chemical. The chemical consisted of several substances of which four (acetone, acrylates, butyl acetate and carbon disulfide) has been previously reported to induce hyposmia or anosmia in workers. Other aetiologies were clinically excluded but a large arachnoidea cyst in the frontal part of the left temporobasal fossa with possible compression of the left entorhinal cortex. The toxic aetiology of anosmia is supported by the acute onset and the temporal relationship with occupational exposure. The silent cyst as the cause of anosmia is improbable, but it may have had some contributory role. Our case illustrates both the challenges when clinically examining patients with work related olfactory impairment and the importance of multi-disciplinary approach to such patients. PMID- 15757993 TI - Occupational molybdenum exposure and a gouty electrician. AB - BACKGROUND: Molybdenum is an essential trace element and a component of xanthine oxidase, which catalyses the formation of urate. The toxicity of molybdenum in humans is considered to be low, but hyperuricaemia and gout-like symptoms have been observed sporadically. METHODS: A case of hyperuricaemia and gouty arthritis in a young man with occupational exposure to molybdenum is described. Improvement during an exposure-free period was followed by a relapse after a reconstruction designed to quantify his molybdenum exposure. CONCLUSION: This case seems to represent the first observation of gout associated with occupational molybdenum exposure, but the association might also be entirely circumstantial. PMID- 15757994 TI - Clustered outbreak of skin and eye complaints among catering staff. AB - In August 2002, kitchen staff at a hotel in Central Scotland experienced skin and eye problems believed to be related to their working environment. Of a total of 20 staff, eight cooks reported problems with a painful red skin affecting the face, eyelids, side and front of neck as well as burning, gritty eyes. Five of the affected individuals were clinically assessed in April 2003. The overall clinical impression was of conjunctivitis and sunburn-like erythema. Examination of the data sheets of all cleaning agents and sprays used within the kitchen pointed against an environmental phototoxin. The kitchen area was inspected and two electric fly killers positioned on the ceiling and sidewalls were found to be incorrectly fitted with UVC tubes. The output of these tubes was spectroradiometrically assessed. The recommended unprotected skin and eye exposure limit was reached in 14 s at a distance of 30 cm from the tubes. An exposure of about 60 s would be sufficient to induce minimal erythema in someone of skin type I/II. These results demonstrate the importance of exposure to ultraviolet radiation as a possible cause of facial erythema and conjunctivitis, no matter how unlikely this may seem. It is recommended that there should be increased awareness of the need to fit the correct type of lamps to electric fly killers and other devices that incorporate UV lamps. PMID- 15757995 TI - Mortality in an Australian cohort of textile workers. PMID- 15757996 TI - Occupational exposure to endotoxin reduces lung cancer rates. PMID- 15757997 TI - Training in occupational medicine. PMID- 15757998 TI - Monitor. PMID- 15757999 TI - Pocketome via comprehensive identification and classification of ligand binding envelopes. AB - We developed a new computational algorithm for the accurate identification of ligand binding envelopes rather than surface binding sites. We performed a large scale classification of the identified envelopes according to their shape and physicochemical properties. The predicting algorithm, called PocketFinder, uses a transformation of the Lennard-Jones potential calculated from a three-dimensional protein structure and does not require any knowledge about a potential ligand molecule. We validated this algorithm using two systematically collected data sets of ligand binding pockets from complexed (bound) and uncomplexed (apo) structures from the Protein Data Bank, 5616 and 11,510, respectively. As many as 96.8% of experimental binding sites were predicted at better than 50% overlap level. Furthermore 95.0% of the asserted sites from the apo receptors were predicted at the same level. We demonstrate that conformational differences between the apo and bound pockets do not dramatically affect the prediction results. The algorithm can be used to predict ligand binding pockets of uncharacterized protein structures, suggest new allosteric pockets, evaluate feasibility of protein-protein interaction inhibition, and prioritize molecular targets. Finally the data base of the known and predicted binding pockets for the human proteome structures, the human pocketome, was collected and classified. The pocketome can be used for rapid evaluation of possible binding partners of a given chemical compound. PMID- 15758001 TI - Concurrent antiplatelet and fibrinolytic therapy. PMID- 15758000 TI - Addition of clopidogrel to aspirin and fibrinolytic therapy for myocardial infarction with ST-segment elevation. AB - BACKGROUND: A substantial proportion of patients receiving fibrinolytic therapy for myocardial infarction with ST-segment elevation have inadequate reperfusion or reocclusion of the infarct-related artery, leading to an increased risk of complications and death. METHODS: We enrolled 3491 patients, 18 to 75 years of age, who presented within 12 hours after the onset of an ST-elevation myocardial infarction and randomly assigned them to receive clopidogrel (300-mg loading dose, followed by 75 mg once daily) or placebo. Patients received a fibrinolytic agent, aspirin, and when appropriate, heparin (dispensed according to body weight) and were scheduled to undergo angiography 48 to 192 hours after the start of study medication. The primary efficacy end point was a composite of an occluded infarct-related artery (defined by a Thrombolysis in Myocardial Infarction flow grade of 0 or 1) on angiography or death or recurrent myocardial infarction before angiography. RESULTS: The rates of the primary efficacy end point were 21.7 percent in the placebo group and 15.0 percent in the clopidogrel group, representing an absolute reduction of 6.7 percentage points in the rate and a 36 percent reduction in the odds of the end point with clopidogrel therapy (95 percent confidence interval, 24 to 47 percent; P<0.001). By 30 days, clopidogrel therapy reduced the odds of the composite end point of death from cardiovascular causes, recurrent myocardial infarction, or recurrent ischemia leading to the need for urgent revascularization by 20 percent (from 14.1 to 11.6 percent, P=0.03). The rates of major bleeding and intracranial hemorrhage were similar in the two groups. CONCLUSIONS: In patients 75 years of age or younger who have myocardial infarction with ST-segment elevation and who receive aspirin and a standard fibrinolytic regimen, the addition of clopidogrel improves the patency rate of the infarct-related artery and reduces ischemic complications. PMID- 15758002 TI - Images in clinical medicine. Marjolin's ulcer. PMID- 15758003 TI - The Groningen protocol--euthanasia in severely ill newborns. PMID- 15758004 TI - Tsunami in Thailand--disaster management in a district hospital. PMID- 15758005 TI - The public health emergency in indonesia--one patient at a time. PMID- 15758006 TI - The accidental addict. PMID- 15758007 TI - Electronic alerts to prevent venous thromboembolism among hospitalized patients. AB - BACKGROUND: Prophylaxis against deep-vein thrombosis in hospitalized patients remains underused. We hypothesized that the use of a computer-alert program to encourage prophylaxis might reduce the frequency of deep-vein thrombosis among high-risk hospitalized patients. METHODS: We developed a computer program linked to the patient database to identify consecutive hospitalized patients at risk for deep-vein thrombosis in the absence of prophylaxis. The program used medical record numbers to randomly assign 1255 eligible patients to an intervention group, in which the responsible physician was alerted to a patient's risk of deep vein thrombosis, and 1251 patients to a control group, in which no alert was issued. The physician was required to acknowledge the alert and could then withhold or order prophylaxis, including graduated compression stockings, pneumatic compression boots, unfractionated heparin, low-molecular-weight heparin, or warfarin. The primary end point was clinically diagnosed, objectively confirmed deep-vein thrombosis or pulmonary embolism at 90 days. RESULTS: More patients in the intervention group than in the control group received mechanical prophylaxis (10.0 percent vs. 1.5 percent, P<0.001) or pharmacologic prophylaxis (23.6 percent vs. 13.0 percent, P<0.001). The primary end point occurred in 61 patients (4.9 percent) in the intervention group, as compared with 103 (8.2 percent) in the control group; the Kaplan-Meier estimates of the likelihood of freedom from deep-vein thrombosis or pulmonary embolism at 90 days were 94.1 percent (95 percent confidence interval, 92.5 to 95.4 percent) and 90.6 percent (95 percent confidence interval, 88.7 to 92.2 percent), respectively (P<0.001). The computer alert reduced the risk of deep-vein thrombosis or pulmonary embolism at 90 days by 41 percent (hazard ratio, 0.59; 95 percent confidence interval, 0.43 to 0.81; P=0.001). CONCLUSIONS: The institution of a computer-alert program increased physicians' use of prophylaxis and markedly reduced the rates of deep vein thrombosis and pulmonary embolism among hospitalized patients at risk. PMID- 15758008 TI - Treatment of early childhood medulloblastoma by postoperative chemotherapy alone. AB - BACKGROUND: The prognosis for young children with medulloblastoma is poor, and survivors are at high risk for cognitive deficits. We conducted a trial of the treatment of this brain tumor by intensive postoperative chemotherapy alone. METHODS: After surgery, children received three cycles of intravenous chemotherapy (cyclophosphamide, vincristine, methotrexate, carboplatin, and etoposide) and intraventricular methotrexate. Treatment was terminated if a complete remission was achieved. Leukoencephalopathy and cognitive deficits were evaluated. RESULTS: Forty-three children were treated according to protocol. In children who had complete resection (17 patients), residual tumor (14), and macroscopic metastases (12), the five-year progression-free and overall survival rates (+/-SE) were 82+/-9 percent and 93+/-6 percent, 50+/-13 percent and 56+/-14 percent, and 33+/-14 percent and 38+/-15 percent, respectively. The rates in 31 patients without macroscopic metastases were 68+/-8 percent and 77+/-8 percent. Desmoplastic histology, metastatic disease, and an age younger than two years were independent prognostic factors for tumor relapse and survival. Treatment strategies at relapse were successful in 8 of 16 patients. There were no major instances of unexpected toxicity. In 19 of 23 children, asymptomatic leukoencephalopathy was detected by magnetic resonance imaging. After treatment, the mean IQ was significantly lower than that of healthy controls within the same age group but higher than that of patients in a previous trial who had received radiotherapy. CONCLUSIONS: Postoperative chemotherapy alone is a promising treatment for medulloblastoma in young children without metastases. PMID- 15758009 TI - Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. AB - BACKGROUND: Glioblastoma, the most common primary brain tumor in adults, is usually rapidly fatal. The current standard of care for newly diagnosed glioblastoma is surgical resection to the extent feasible, followed by adjuvant radiotherapy. In this trial we compared radiotherapy alone with radiotherapy plus temozolomide, given concomitantly with and after radiotherapy, in terms of efficacy and safety. METHODS: Patients with newly diagnosed, histologically confirmed glioblastoma were randomly assigned to receive radiotherapy alone (fractionated focal irradiation in daily fractions of 2 Gy given 5 days per week for 6 weeks, for a total of 60 Gy) or radiotherapy plus continuous daily temozolomide (75 mg per square meter of body-surface area per day, 7 days per week from the first to the last day of radiotherapy), followed by six cycles of adjuvant temozolomide (150 to 200 mg per square meter for 5 days during each 28 day cycle). The primary end point was overall survival. RESULTS: A total of 573 patients from 85 centers underwent randomization. The median age was 56 years, and 84 percent of patients had undergone debulking surgery. At a median follow-up of 28 months, the median survival was 14.6 months with radiotherapy plus temozolomide and 12.1 months with radiotherapy alone. The unadjusted hazard ratio for death in the radiotherapy-plus-temozolomide group was 0.63 (95 percent confidence interval, 0.52 to 0.75; P<0.001 by the log-rank test). The two-year survival rate was 26.5 percent with radiotherapy plus temozolomide and 10.4 percent with radiotherapy alone. Concomitant treatment with radiotherapy plus temozolomide resulted in grade 3 or 4 hematologic toxic effects in 7 percent of patients. CONCLUSIONS: The addition of temozolomide to radiotherapy for newly diagnosed glioblastoma resulted in a clinically meaningful and statistically significant survival benefit with minimal additional toxicity. PMID- 15758010 TI - MGMT gene silencing and benefit from temozolomide in glioblastoma. AB - BACKGROUND: Epigenetic silencing of the MGMT (O6-methylguanine-DNA methyltransferase) DNA-repair gene by promoter methylation compromises DNA repair and has been associated with longer survival in patients with glioblastoma who receive alkylating agents. METHODS: We tested the relationship between MGMT silencing in the tumor and the survival of patients who were enrolled in a randomized trial comparing radiotherapy alone with radiotherapy combined with concomitant and adjuvant treatment with temozolomide. The methylation status of the MGMT promoter was determined by methylation-specific polymerase-chain reaction analysis. RESULTS: The MGMT promoter was methylated in 45 percent of 206 assessable cases. Irrespective of treatment, MGMT promoter methylation was an independent favorable prognostic factor (P<0.001 by the log-rank test; hazard ratio, 0.45; 95 percent confidence interval, 0.32 to 0.61). Among patients whose tumor contained a methylated MGMT promoter, a survival benefit was observed in patients treated with temozolomide and radiotherapy; their median survival was 21.7 months (95 percent confidence interval, 17.4 to 30.4), as compared with 15.3 months (95 percent confidence interval, 13.0 to 20.9) among those who were assigned to only radiotherapy (P=0.007 by the log-rank test). In the absence of methylation of the MGMT promoter, there was a smaller and statistically insignificant difference in survival between the treatment groups. CONCLUSIONS: Patients with glioblastoma containing a methylated MGMT promoter benefited from temozolomide, whereas those who did not have a methylated MGMT promoter did not have such a benefit. PMID- 15758011 TI - Clinical practice. Neurocardiogenic syncope. PMID- 15758012 TI - Anemia of chronic disease. PMID- 15758013 TI - Images in clinical medicine. Mondor's disease. PMID- 15758014 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 7-2005. A 59-year-old woman with an incidentally discovered adrenal nodule. PMID- 15758015 TI - Electronic medical alerts--so simple, so complex. PMID- 15758016 TI - Chemotherapy for brain tumors--a new beginning. PMID- 15758017 TI - Implantable cardioverter-defibrillator therapy after myocardial infarction. PMID- 15758018 TI - Isosorbide dinitrate and hydralazine in blacks with heart failure. PMID- 15758019 TI - Intradermal vaccination against influenza. PMID- 15758020 TI - The U.S. vaccine supply. PMID- 15758021 TI - Medical mystery answer: male infant with unusual weight gain and rash. PMID- 15758022 TI - Peripartum dissection of the right coronary artery. PMID- 15758023 TI - Imatinib and regression of type 2 diabetes. PMID- 15758024 TI - Myosin-1a is critical for normal brush border structure and composition. AB - To develop our understanding of myosin-1a function in vivo, we have created a mouse line null for the myosin-1a gene. Myosin-1a knockout mice demonstrate no overt phenotypes at the whole animal level but exhibit significant perturbations and signs of stress at the cellular level. Among these are defects in microvillar membrane morphology, distinct changes in brush-border organization, loss of numerous cytoskeletal and membrane components from the brush border, and redistribution of intermediate filament proteins into the brush border. We also observed significant ectopic recruitment of another short-tailed class I motor, myosin-1c, into the brush border of knockout enterocytes. This latter finding, a clear demonstration of functional redundancy among vertebrate myosins-I, may account for the lack of a whole animal phenotype. Nevertheless, these results indicate that myosin-1a is a critical multifunctional component of the enterocyte, required for maintaining the normal composition and highly ordered structure of the brush border. PMID- 15758025 TI - The aftiphilin/p200/gamma-synergin complex. AB - Aftiphilin is a protein that was recently identified in database searches for proteins with motifs that interact with AP-1 and clathrin, but its function is unknown. Here we demonstrate that aftiphilin has a second, atypical clathrin binding site, YQW, that colocalizes with AP-1 by immunofluorescence, and that is enriched in clathrin-coated vesicles (CCVs), confirming that it is a bona fide component of the CCV machinery. By gel filtration, aftiphilin coelutes with two other AP-1 binding partners, p200a and gamma-synergin. Antibodies against any one of these three proteins immunoprecipitate the other two, and knocking down any of the three proteins by siRNA causes a reduction in the levels of the other two, indicating that they form a stable complex. Like AP-1-depleted cells, aftiphilin depleted cells missort a CD8-furin chimera and the lysosomal enzyme cathepsin D. However, whereas AP-1-depleted cells recycle endocytosed transferrin more slowly than untreated cells, aftiphilin-depleted cells accumulate endocytosed transferrin in a peripheral compartment and recycle it more rapidly. These observations show that in general, the aftiphilin/p200/gamma-synergin complex facilitates AP-1 function, but the complex may have additional functions as well, because of the opposing effects of the two knockdowns on transferrin recycling. PMID- 15758026 TI - High mobility of flap endonuclease 1 and DNA polymerase eta associated with replication foci in mammalian S-phase nucleus. AB - Originally detected in fixed cells, DNA replication foci (RFi) were later visualized in living cells by using green fluorescent protein (GFP)-tagged proliferating cell nuclear antigen (PCNA) and DNA ligase I. It was shown using fluorescence redistribution after photobleaching (FRAP) assay that focal GFP-PCNA slowly exchanged, suggesting the existence of a stable replication holocomplex. Here, we used the FRAP assay to study the dynamics of the GFP-tagged PCNA-binding proteins: Flap endonuclease 1 (Fen1) and DNA polymerase eta (Pol eta). We also used the GFP-Cockayne syndrome group A (CSA) protein, which does associate with transcription foci after DNA damage. In normal cells, GFP-Pol eta and GFP-Fen1 are mobile with residence times at RFi (t(m)) approximately 2 and approximately 0.8 s, respectively. GFP-CSA is also mobile but does not concentrate at discrete foci. After methyl methanesulfonate (MMS) damage, the mobile fraction of focal GFP-Fen1 decreased and t(m) increased, but it then recovered. The mobilities of focal GFP-Pol eta and GFP-PCNA did not change after MMS. The mobility of GFP-CSA did not change after UV-irradiation. These data indicate that the normal replication complex contains at least two mobile subunits. The decrease of the mobile fraction of focal GFP-Fen1 after DNA damage suggests that Fen1 exchange depends on the rate of movement of replication forks. PMID- 15758028 TI - Homeostatic adjustment and metabolic remodeling in glucose-limited yeast cultures. AB - We studied the physiological response to glucose limitation in batch and steady state (chemostat) cultures of Saccharomyces cerevisiae by following global patterns of gene expression. Glucose-limited batch cultures of yeast go through two sequential exponential growth phases, beginning with a largely fermentative phase, followed by an essentially completely aerobic use of residual glucose and evolved ethanol. Judging from the patterns of gene expression, the state of the cells growing at steady state in glucose-limited chemostats corresponds most closely with the state of cells in batch cultures just before they undergo this "diauxic shift." Essentially the same pattern was found between chemostats having a fivefold difference in steady-state growth rate (the lower rate approximating that of the second phase respiratory growth rate in batch cultures). Although in both cases the cells in the chemostat consumed most of the glucose, in neither case did they seem to be metabolizing it primarily through respiration. Although there was some indication of a modest oxidative stress response, the chemostat cultures did not exhibit the massive environmental stress response associated with starvation that also is observed, at least in part, during the diauxic shift in batch cultures. We conclude that despite the theoretical possibility of a switch to fully aerobic metabolism of glucose in the chemostat under conditions of glucose scarcity, homeostatic mechanisms are able to carry out metabolic adjustment as if fermentation of the glucose is the preferred option until the glucose is entirely depleted. These results suggest that some aspect of actual starvation, possibly a component of the stress response, may be required for triggering the metabolic remodeling associated with the diauxic shift. PMID- 15758027 TI - Dynamic sorting of nuclear components into distinct nucleolar caps during transcriptional inhibition. AB - Nucleolar segregation is observed under some physiological conditions of transcriptional arrest. This process can be mimicked by transcriptional arrest after actinomycin D treatment leading to the segregation of nucleolar components and the formation of unique structures termed nucleolar caps surrounding a central body. These nucleolar caps have been proposed to arise from the segregation of nucleolar components. We show that contrary to prevailing notion, a group of nucleoplasmic proteins, mostly RNA binding proteins, relocalized from the nucleoplasm to a specific nucleolar cap during transcriptional inhibition. For instance, an exclusively nucleoplasmic protein, the splicing factor PSF, localized to nucleolar caps under these conditions. This structure also contained pre-rRNA transcripts, but other caps contained either nucleolar proteins, PML, or Cajal body proteins and in addition nucleolar or Cajal body RNAs. In contrast to the capping of the nucleoplasmic components, nucleolar granular component proteins dispersed into the nucleoplasm, although at least two (p14/ARF and MRP RNA) were retained in the central body. The nucleolar caps are dynamic structures as determined using photobleaching and require energy for their formation. These findings demonstrate that the process of nucleolar segregation and capping involves energy-dependent repositioning of nuclear proteins and RNAs and emphasize the dynamic characteristics of nuclear domain formation in response to cellular stress. PMID- 15758031 TI - Altered nerve excitability properties in established diabetic neuropathy. AB - The underlying cause of diabetic neuropathy remains unclear, although pathological studies have suggested an ischaemic basis related to microangiopathy, possibly mediated through effects on the energy-dependent Na+/K+ pump. To investigate the pathophysiology of diabetic neuropathy, axonal excitability techniques were undertaken in 20 diabetic patients with neuropathy severity graded through a combination of quantitative sensory testing (QST) using a vibratory stimulus, assessment of symptom severity using the Total Neuropathy Symptom Score (T-NSS) and measurement of glycosylated haemoglobin as a marker of disease control. To assess axonal excitability, compound muscle action potentials were recorded at rest from abductor pollicis brevis following stimulation of the median nerve, and stimulus-response behaviour, threshold electrotonus, a current threshold relationship and the recovery of excitability were recorded in each patient. All patients had established neuropathy, with abnormalities of T-NSS present in all patients and QST abnormalities present in 65%. Compared with controls, diabetic neuropathy patients had significant reduction in maximal CMAP amplitude (P < 0.0005), accompanied by a 'fanning in' of threshold electrotonus. In addition, the strength-duration time constant was decreased in diabetic neuropathy patients and recovery cycles were altered with reductions in refractoriness, the duration of the relative refractory period, superexcitability and subexcitability. It is proposed that while the changes in threshold electrotonus with supportive findings in the current-threshold relationship are consistent with axonal depolarization, possibly mediated by a decrease in Na+/K+ pump activity, the alterations in the recovery cycle of excitability could be explained on the basis of a smaller action potential, reflecting a limitation on the nodal driving current imposed by a reduction in Na+ conductances. PMID- 15758029 TI - Initiation of embryonic cardiac pacemaker activity by inositol 1,4,5 trisphosphate-dependent calcium signaling. AB - In the adult, the heart rate is driven by spontaneous and repetitive depolarizations of pacemaker cells to generate a firing of action potentials propagating along the conduction system and spreading into the ventricles. In the early embryo before E9.5, the pacemaker ionic channel responsible for the spontaneous depolarization of cells is not yet functional. Thus the mechanisms that initiate early heart rhythm during cardiogenesis are puzzling. In the absence of a functional pacemaker ionic channel, the oscillatory nature of inositol 1,4,5-trisphosphate (InsP3)-induced intracellular Ca2+ signaling could provide an alternative pacemaking mechanism. To test this hypothesis, we have engineered pacemaker cells from embryonic stem (ES) cells, a model that faithfully recapitulates early stages of heart development. We show that InsP3 dependent shuttle of free Ca2+ in and out of the endoplasmic reticulum is essential for a proper generation of pacemaker activity during early cardiogenesis and fetal life. PMID- 15758032 TI - Quantitative post-mortem study of the hippocampus in chronic epilepsy: seizures do not inevitably cause neuronal loss. AB - Hippocampal sclerosis describes a pattern of neuronal loss and gliosis involving the medial temporal structures most often encountered in patients with epilepsy. It is still a matter for debate as to whether this lesion is acquired during the course of the patient's seizure history or is present at the outset. Early febrile seizures, episodes of status epilepticus as well as repetitive brief seizures may all contribute to the evolution of hippocampal sclerosis. In addition, genetic factors and developmental abnormalities of the hippocampus may both increase vulnerability to seizures and hippocampal injury. Recent human studies have addressed neuropathological changes in young adults and children undergoing surgery for refractory seizures with hippocampal sclerosis. Post mortem examination, however, provides the opportunity to evaluate the effect of a lifetime of seizures on both left and right hippocampi, and the presence of any co-existing malformation. Post-mortem stereological analysis of 28 patients with poorly controlled seizures has confirmed a subgroup with absence of significant hippocampal neuronal loss despite decades of generalized seizures, including status epilepticus. The presence of granule cell dispersion correlated to the severity of hippocampal neuronal loss. Furthermore, in patients with confirmed hippocampal sclerosis at post-mortem examination, stereological assessment of the neocortex failed to confirm significant white matter neuronal heterotopia that might indicate an underlying developmental abnormality. In conclusion, seizures do not invariably lead to hippocampal injury and white matter heterotopia is not invariably associated with hippocampal sclerosis. PMID- 15758030 TI - Cisternal rab proteins regulate Golgi apparatus redistribution in response to hypotonic stress. AB - We show that a physiological role of the extensively studied cisternal Golgi rab protein, rab6, is modulation of Golgi apparatus response to stress. Taking exposure of cells to hypotonic media as the best-known example of mammalian Golgi stress response, we found that hypotonic-induced tubule extension from the Golgi apparatus was sensitive to GDP-rab6a expression. Similarly, we found that Golgi tubulation induced by brefeldin A, a known microtubule-dependent process, was inhibited by GDP-restricted rab6a, rab6a', and rab33b, the most commonly studied cisternal rab proteins. These GDP-rab levels were sufficient to inhibit rab induced redistribution of Golgi glycosyltransferases into the endoplasmic reticulum (ER), also a microtubule-dependent process, and to depress Golgi membrane association of the GTP-conformer of rab6. Nocodazole-induced Golgi scattering, a microtubule-independent process, also was inhibited by GDP-rab6a expression. In comparison, we found similar GDP-rab expression levels had little inhibitory effect on another microtubule-independent process, constitutive recycling of Golgi resident proteins to the ER. We conclude that Golgi cisternal rabs, and in particular rab6a, are regulators of the Golgi response to stress and presumably the molecular targets of stress-activated signaling pathway(s). Moreover, we conclude that rab6a can regulate select microtubule-independent processes as well as microtubule-dependent processes. PMID- 15758033 TI - The role of the human thalamus in processing corollary discharge. AB - Corollary discharge signals play an important role in monitoring self-generated movements to guarantee spatial constancy. Recent work in macaques suggests that the thalamus conveys corollary discharge information of upcoming saccades passing from the superior colliculus to the frontal eye field. The present study aimed to investigate the involvement of the thalamus in humans by assessing the effect of thalamic lesions on the processing of corollary discharge information. Thirteen patients with selective thalamic lesions and 13 healthy age-matched control subjects performed a saccadic double-step task in which retino-spatial dissonance was induced, i.e. the retinal vector of the second target and the movement vector of the second saccade were different. Thus, the subjects could not rely on retinal information alone, but had to use corollary discharge information to correctly perform the second saccade. The amplitudes of first and second saccades were significantly smaller in patients than in controls. Five thalamic lesion patients showed unilateral deficits in using corollary discharge information, as revealed by asymmetries compared with the other patients and controls. Three patients with lateral thalamic lesions including the ventrolateral nucleus (VL) were impaired contralaterally to the side of damage and one patient with a lesion in the mediodorsal thalamus (MD) was impaired ipsilaterally to the lesion. The largest asymmetry was found in a patient with a bilateral thalamic lesion. The results provide evidence for a thalamic involvement in the processing of corollary discharge information in humans, with a potential role of both the VL and MD nuclei. PMID- 15758034 TI - Cigarette smoking and the progression of multiple sclerosis. AB - An increased risk of multiple sclerosis among smokers has been found in several prospective epidemiological studies. The association between smoking and progression of multiple sclerosis has not been examined. We identified patients who had a first multiple sclerosis diagnosis recorded in the General Practice Research Database (GPRD) between January 1993 and December 2000. Their diagnosis and date of first symptoms were confirmed through examination of medical records. Smoking status was obtained from the computer records. To assess the association between smoking and risk of multiple sclerosis, we conducted a case-control study nested in the GPRD. Up to 10 controls per case were randomly selected, matched on age, sex, practice, date of joining the practice and availability of smoking data. To assess the association between smoking and progression of multiple sclerosis, we conducted a cohort study of multiple sclerosis cases with a relapsing-remitting onset. Our nested case-control study included 201 cases of multiple sclerosis and 1913 controls. The odds ratio [95% confidence interval (CI)] of multiple sclerosis was 1.3 (1.0-1.7) for ever smokers compared with never smokers. Our cohort study included 179 cases with a mean (median) length of follow-up of 5.3 (5.3) years. The hazard ratio of secondary progression was 3.6 (95% CI 1.3-9.9) for ever smokers compared with never smokers. These results support the hypothesis that cigarette smoking is associated with an increased risk of multiple sclerosis, and suggest that smoking may be a risk factor for transforming a relapsing-remitting clinical course into a secondary progressive course. PMID- 15758035 TI - Seizure-related short-term plasticity of benzodiazepine receptors in partial epilepsy: a [11C]flumazenil-PET study. AB - We have undertaken a test-re-test [11C]flumazenil (FMZ) PET study in 10 drug resistant epileptic patients, including six with a mesiotemporal epilepsy (MTE), and 10 normal controls, in order to investigate seizure-related short-term plasticity of benzodiazepine (BZD) receptors. All subjects underwent two FMZ-PET scans at a 1 week interval. Patients benefited from a concurrent video-EEG monitoring which allowed determination of the duration of the interictal period (IP) preceding each PET. Test-re-test whole brain B'(max) variations, evaluated with a partial-saturation injection protocol, were similarly observed in patients and controls, suggesting a physiological modulation of BZD receptors. Five patients (50%), but no controls, also demonstrated clinically significant test-re test FMZ-PET variations in the mesial temporal region. This was observed in all three patients with MTE and no hippocampal atrophy in whom only the PET study associated with the shortest IP correctly identified the epileptogenic zone. Statistical analysis revealed a significant effect of IP duration on BZD receptor B'(max) in MTE patients, suggesting that the shorter the IP, the lower the B'(max) in the epileptogenic hippocampus. FMZ-PET appears to be an interesting tool for investigating both normal and abnormal short-term modulations of the BZD receptor system, and should ideally be performed within a few days following a seizure in patients with MTE and a normal MRI. PMID- 15758036 TI - Evidence of elevated glutamate in multiple sclerosis using magnetic resonance spectroscopy at 3 T. AB - Histopathological reports of multiple sclerosis and its animal models have shown evidence of a link between axonal injury in active lesions and impaired glutamate metabolism. Mature oligodendrocytes play a role in glutamate uptake to maintain glutamate homeostasis but in multiple sclerosis white matter the loss of expression of glutamate transporters in the lesion vicinity results in ineffective glutamate removal. Using a magnetic resonance spectroscopy technique that isolates the glutamate resonance at 3 T, we compared glutamate levels between normal subjects and multiple sclerosis patients in different brain areas. Metabolite concentrations (glutamate, glutamine, N-acetyl-aspartate, myo inositol, choline, creatine) were derived from LCmodel and corrected for T1 relaxation time. Glutamate concentrations were found to be elevated in acute lesions (P = 0.02) and normal-appearing white matter (P = 0.03), with no significant elevation in chronic lesions (P = 0.77). The N-acetyl-aspartate level in chronic lesions was significantly lower (P < 0.001) than in acute lesions and normal-appearing white matter. The choline level in acute lesions was significantly higher (P < 0.001) than in chronic lesions. Evidence was also found for increased glial activity in multiple sclerosis, with significantly higher (P < 0.001) myo-inositol levels in acute lesions compared with control white matter. These in vivo results support the hypothesis that altered glutamate metabolism is present in brains of multiple sclerosis patients. PMID- 15758037 TI - Causes of cross-over in unilateral neglect: between-group comparisons, within patient dissociations and eye movements. AB - Patients with left unilateral neglect bisect long horizontal lines to the right of the true centre. However, when given short lines, many of the same patients mark the midpoint to the left of the true centre, towards the otherwise neglected space. This paradoxical phenomenon has been termed 'cross-over' and is difficult to explain based on current accounts of the neglect syndrome. To explore the causes of cross-over, in a first study we evaluated bisection of 20, 100 and 200 mm horizontal lines in groups of unilateral brain-damaged patients with neglect and hemianopia, with neglect and no hemianopia, with hemianopia and no neglect and without neglect or hemianopia. Cross-over of 20 mm lines was found only in neglect patients with hemianopia. To ascertain further the influence of visual field defects on cross-over, in a second study we compared the performance of two right-brain-damaged patients with contralesional neglect and inferior quadrantanopia with that of a patient with inferior quadrantanopia and no neglect. Patients bisected lines oriented so as to cross or uncross the blind quadrant of the visual field. When short 20 mm lines crossed the blind quadrant, neglect patients showed cross-over; when the same lines crossed the seeing quadrants cross-over was absent. These findings were confirmed by the examination of a neglect patient with sparing of the central 5 degrees of the contralesional left visual hemifield in the right eye and no sparing in the left eye. In monocular viewing, cross-over was present when 20 mm lines were bisected with the left eye and absent when bisected with the right eye. Recording of eye movements showed that at the moment of bisection left eye fixations shifted towards the contralesional line endpoint whereas right eye fixations remained anchored to the centre of the line. With long lines, both eyes deviated ipsilesionally. These results show that in neglect patients ipsilesional deviation in the bisection of long lines turns into apparently paradoxical contralesional bisection of short ones only when these cross a retinotopically blind sector of the neglected space. Cross-over seems to depend on the small spatial effects produced by reflexive contralesional gaze shifts allowing eccentric fixations with the seeing hemifield. During the bisection of long lines, these effects are cancelled out by the strong attentional deviation induced by the marked extension of the ipsilesional line segment. This explanation establishes coherence between cross over and current accounts of the neglect syndrome. PMID- 15758038 TI - Long-term seizure outcomes following epilepsy surgery: a systematic review and meta-analysis. AB - Assessment of long-term outcomes is essential in brain surgery for epilepsy, which is an irreversible intervention for a chronic condition. Excellent short term results of resective epilepsy surgery have been established, but less is known about long-term outcomes. We performed a systematic review and meta analysis of the evidence on this topic. To provide evidence-based estimates of long-term results of various types of epilepsy surgery and to identify sources of variation in results of published studies, we searched Medline, Index Medicus, the Cochrane database, bibliographies of reviews, original articles and book chapters to identify articles published since 1991 that contained > or =20 patients of any age, undergoing resective or non-resective epilepsy surgery, and followed for a mean/median of > or =5 years. Two reviewers independently assessed study eligibility and extracted data, resolving disagreements through discussion. Seventy-six articles fulfilled our eligibility criteria, of which 71 reported on resective surgery (93%) and five (7%) on non-resective surgery. There were no randomized trials and only six studies had a control group. Some articles contributed more than one study, yielding 83 studies of which 78 dealt with resective surgery and five with non-resective surgery. Forty studies (51%) of resective surgery referred to temporal lobe surgery, 25 (32%) to grouped temporal and extratemporal surgery, seven (9%) to frontal surgery, two (3%) to grouped extratemporal surgery, two (3%) to hemispherectomy, and one (1%) each to parietal and occipital surgery. In the non-resective category, three studies reported outcomes after callosotomy and two after multiple subpial transections. The median proportion of long-term seizure-free patients was 66% with temporal lobe resections, 46% with occipital and parietal resections, and 27% with frontal lobe resections. In the long term, only 35% of patients with callosotomy were free of most disabling seizures, and 16% with multiple subpial transections remained free of all seizures. The year of operation, duration of follow-up and outcome classification system were most strongly associated with outcomes. Almost all long-term outcome studies describe patient cohorts without controls. Although there is substantial variation in outcome definition and methodology among the studies, consistent patterns of results emerge for various surgical interventions after adjusting for sources of heterogeneity. The long-term (> or =5 years) seizure free rate following temporal lobe resective surgery was similar to that reported in short-term controlled studies. On the other hand, long-term seizure freedom was consistently lower after extratemporal surgery and palliative procedures. PMID- 15758039 TI - Neural basis of eye gaze processing deficits in autism. AB - Impairments in using eye gaze to establish joint attention and to comprehend the mental states and intentions of other people are striking features of autism. Here, using event-related functional MRI (fMRI), we show that in autism, brain regions involved in gaze processing, including the superior temporal sulcus (STS) region, are not sensitive to intentions conveyed by observed gaze shifts. On congruent trials, subjects watched as a virtual actor looked towards a checkerboard that appeared in her visual field, confirming the subject's expectation regarding what the actor 'ought to do' in this context. On incongruent trials, she looked towards empty space, violating the subject's expectation. Consistent with a prior report from our laboratory that used this task in neurologically normal subjects, 'errors' (incongruent trials) evoked more activity in the STS and other brain regions linked to social cognition, indicating a strong effect of intention in typically developing subjects (n = 9). The same brain regions were activated during observation of gaze shifts in subjects with autism (n = 10), but did not differentiate congruent and incongruent trials, indicating that activity in these regions was not modulated by the context of the perceived gaze shift. These results demonstrate a difference in the response of brain regions underlying eye gaze processing in autism. We conclude that lack of modulation of the STS region by gaze shifts that convey different intentions contributes to the eye gaze processing deficits associated with autism. PMID- 15758040 TI - Adhesion of human and mouse platelets to collagen under shear: a unifying model. AB - There is presently confusion as to the roles of alpha2beta1 and GPVI in supporting platelet adhesion and aggregate formation on collagen at intermediate/high shear. Recent studies have reported essential, partial, or dispensable roles for either receptor in supporting these events, and the possibility that there may be fundamental differences between their roles in human and mouse platelets has been proposed. Further, the recent recognition that Src family tyrosine kinases contribute to signaling by alpha2beta1 and other adhesive receptors, in addition to GPVI, has added to this debate. The present study compares the roles of alpha2beta1, GPVI, and Src-dependent kinases in supporting adhesion and aggregation in human and mouse platelets in whole blood using blocking antibodies, mutant mice, and a novel inhibitor of Src kinases, PD0173952, which is effective in plasma. The results demonstrate that the fundamental processes of adhesion and aggregate formation are conserved in mice and human platelets and that two mechanisms of stable adhesion and activation on collagen exist. These can be distinguished by the contributions of GPVI and alpha2beta1, with GPVI-mediated platelet activation either preceding or following integrin-mediated adhesion. The relative contribution of each pathway depends on environmental conditions and may also reflect platelet heterogeneity. These observations form the basis of a unifying two-state model of platelet adhesion and aggregate formation on collagen that is conserved between human and mouse platelets. PMID- 15758041 TI - Airway smooth muscle cells enhance C3a-induced mast cell degranulation following cell-cell contact. AB - Growing evidence suggests that anaphylatoxins, C3a and C5a, play important roles in innate immunity and may also participate in the pathogenesis of asthma. Previous studies with animal models and immunohistochemistry analysis of lung tissue indicated that anaphylatoxins may regulate airway hyperresponsiveness (AHR) in asthma via the activation of their cell surface G protein-coupled receptors (C3aR and C5aR) in airway smooth muscle (ASM) cells. Using RT-PCR, flow cytometry, and confocal microscopy, we made the surprising observation that while C3aR and C5aR were expressed in human mast cells, they were not present in cultured primary human or murine ASM cells. Furthermore, we could not detect C3aR in smooth muscle-positive cells of human trachea or bronchus. Interestingly, incubation of human mast cells with ASM cells, but not its culture supernatant, caused a significant enhancement of C3a-induced mast cell degranulation. Although stem cell factor (SCF) and its receptor c-kit are constitutively expressed on ASM cells and mast cells, respectively, neutralizing antibodies to SCF and c-kit failed to inhibit ASM cell-mediated enhancement of mast cell degranulation. However, dexamethasone-treated ASM cells were normal for cell surface SCF expression but were significantly less effective in enhancing C3a-induced mast cell degranulation when compared with untreated cells. These findings suggest that cell-cell interaction between ASM cells and mast cells, via a SCF-c-kit independent but dexamethasone-sensitive mechanism, enhances C3a-induced mast cell degranulation, which likely regulates ASM function, thus contributing to the pathogenesis of asthma. PMID- 15758042 TI - Effect of transition metals (Mn, Cu, Fe) and deoxycholic acid (DA) on the conversion of PrPC to PrPres. AB - The PMCA (protein misfolding cyclic amplification) technique has been shown to drive the amplification of misfolded prion protein by PrP(Sc) seeds during several cycles of incubation-sonication. Here, we report that cyclic amplification of normal hamster brain homogenates treated with a number of transition metals (manganese [Mn], copper [Cu], and iron [Fe]) leads to conversion of PrP(C) into protease-resistant PrP(res). The efficiency of PrP(res) formation and the glycoforms induced by Mn were different from those obtained by Cu and Fe. Previous results have shown higher Mn and lower Cu levels in the affinity-purified PrP(Sc) from the brain of prion diseases compared with normal hamster brain homogenates. We focused on Mn because we observed higher levels of Mn in whole brain, mitochondria, and scrapie-associated fibril-enriched fractions from the brains of animals with prion disease. In the presence of minute quantities of Mn-induced PrP(res) template with a large amount of PrP(C), PrP(res) amplification is observed. A metal chelater, EDTA reverses the effect of Mn on PrP(res) amplification, suggesting that Mn may play a role in the formation of PrP(res). It has been proposed that metal-catalyzed oxidation of PrP leads to the oxidation of amino acids and extensive aggregation of oxidized PrP. Carboxyl acids such as deoxycholic acid (DA) are oxidized molecules produced by 3' oxidation pathway. In in vitro studies, the potent effect of Mn on PrP(res) amplification is augmented by DA in a dose-dependent manner. On the basis of the evidence of the elevated Mn levels in scrapie-associated fibril (SAF)-enriched preparations from the brains of animals with prion disease, Mn-loaded PrP and oxidized molecules such as carboxyl acids may contribute to the formation of the scrapie isoform of PrP in prion diseases. PMID- 15758043 TI - Autocrine production of prostaglandin F2alpha enhances phenotypic transformation of normal rat kidney fibroblasts. AB - We have used normal rat kidney (NRK) fibroblasts as an in vitro model system to study cell transformation. These cells obtain a transformed phenotype upon stimulation with growth-modulating factors such as retinoic acid (RA) or transforming growth factor-beta (TGF-beta). Patch-clamp experiments showed that transformation is paralleled by a profound membrane depolarization from around 70 to -20 mV. This depolarization is caused by a compound in the medium conditioned by transformed NRK cells, which enhances intracellular Ca2+ levels and thereby activates Ca2+-dependent Cl- channels. This compound was identified as prostaglandin F2alpha (PGF2alpha) using electrospray ionization mass spectrometry. The active concentration in the medium conditioned by transformed NRK cells as determined using an enzyme immunoassay was 19.7 +/- 2.5 nM (n = 6), compared with 1.5 +/- 0.1 nM (n = 3) conditioned by nontransformed NRK cells. Externally added PGF2alpha was able to trigger NRK cells that had grown to density arrest to restart their proliferation. This proliferation was inhibited when the FP receptor (i.e., natural receptor for PGF2alpha) was blocked by AL 8810. RA-induced phenotypic transformation of NRK cells was partially (approximately 25%) suppressed by AL-8810. Our results demonstrate that PGF2alpha acts as an autocrine enhancer and paracrine inducer of cell transformation and suggest that it may play a crucial role in carcinogenesis in general. PMID- 15758044 TI - FRET-based voltage probes for confocal imaging: membrane potential oscillations throughout pancreatic islets. AB - Insulin secretion is dependent on coordinated pancreatic islet physiology. In the present study, we found a way to overcome the limitations of cellular electrophysiology to optically determine cell membrane potential (V(m)) throughout an islet by using a fast voltage optical dye pair. Using laser scanning confocal microscopy (LSCM), we observed fluorescence (Forster) resonance energy transfer (FRET) with the fluorescent donor N-(6-chloro-7-hydroxycoumarin-3 carbonyl)-dimyristoylphosphatidyl-ethanolamine and the acceptor bis-(1,3 diethylthiobarbiturate) trimethine oxonol in the plasma membrane of essentially every cell within an islet. The FRET signal was approximately linear from V(m) 70 to +50 mV with a 2.5-fold change in amplitude. We evaluated the responses of islet cells to glucose and tetraethylammonium. Essentially, every responding cell in a mouse islet displayed similar time-dependent changes in V(m). When V(m) was measured simultaneously with intracellular Ca2+, all active cells showed tight coupling of V(m) to islet cell Ca2+ changes. Our findings indicate that FRET based, voltage-sensitive dyes used in conjunction with LSCM imaging could be extremely useful in studies of excitation-secretion coupling in intact islets of Langerhans. PMID- 15758045 TI - RF-2 gene modulates proteinuria and albuminuria independently of changes in glomerular permeability in the fawn-hooded hypertensive rat. PMID- 15758046 TI - Genetic engineering of glomerular sclerosis in the mouse via control of onset and severity of podocyte-specific injury. AB - This study aimed to generate a mouse model of acquired glomerular sclerosis. A model system that allows induction of podocyte injury in a manner in which onset and severity can be controlled was designed. A transgenic mouse strain (NEP25) that expresses human CD25 selectively in podocytes was first generated. Injection of anti-Tac (Fv)-PE38 (LMB2), an immunotoxin with specific binding to human CD25, induced progressive nonselective proteinuria, ascites, and edema in NEP25 mice. Podocytes showed foot process effacement, vacuolar degeneration, detachment and downregulation of synaptopodin, WT-1, nephrin, and podocalyxin. Mesangial cells showed matrix expansion, increased collagen, mesangiolysis, and, later, sclerosis. Parietal epithelial cells showed vacuolar degeneration and proliferation, whereas endothelial cells were swollen. The severity of the glomerular injury was LMB2 dose dependent. With 1.25 ng/g body wt or more, NEP25 mice developed progressive glomerular damage and died within 2 wk. With 0.625 ng/g body wt of LMB2, NEP25 mice survived >4 wk and developed focal segmental glomerular sclerosis. Thus, the study has established a mouse model of acquired progressive glomerular sclerosis in which onset and severity can be preprogrammed by experimental maneuvers. PMID- 15758048 TI - Transient disruption of ventrolateral prefrontal cortex during verbal encoding affects subsequent memory performance. AB - Episodic memory supports conscious remembrance of everyday experience. Prior functional neuroimaging data indicate that episodic encoding during phonological task performance is correlated with activation in bilateral posterior ventrolateral prefrontal cortex (pVLPFC), although uncertainty remains regarding whether these prefrontal regions make necessary contributions to episodic memory formation. Using functional MRI data to guide application of single-pulse transcranial magnetic stimulation (spTMS), this study examined the necessity of left and right pVLPFC for episodic encoding (as expressed through subsequent memory performance). To assess the timing of critical computations, pVLPFC function was transiently disrupted at different poststimulus onset times while subjects made syllable decisions about visually presented familiar and unfamiliar words; subsequent memory for these stimuli was measured. Results revealed that left pVLPFC disruption during encoding of familiar words impaired subsequent memory, expressed as a decline in recognition confidence, with disruption being maximal at 380 ms after stimulus onset. In contrast, right pVLPFC disruption facilitated subsequent memory for familiar words, expressed as an increase in medium confidence recognition, with this facilitation being maximal at 380 ms. Finally, phonological (syllable) decision accuracy was facilitated by right pVLPFC disruption, with this effect being maximal at 340 ms, but was unaffected by left pVLPFC disruption. These findings suggest that left pVLPFC mechanisms onset between 300 and 400 ms during phonological processing of words, with these mechanisms appearing necessary for effective episodic encoding. In contrast, disruption of correlated mechanisms in right pVLPFC facilitates encoding, perhaps by inducing a functional shift in the mechanisms engaged during learning. PMID- 15758049 TI - Adaptation at synaptic connections to layer 2/3 pyramidal cells in rat visual cortex. AB - Neocortical synapses express differential dynamic properties. When activated at high frequencies, the amplitudes of the subsequent postsynaptic responses may increase or decrease, depending on the stimulation frequency and on the properties of that particular synapse. Changes in the synaptic dynamics can dramatically affect the communication between nerve cells. Motivated by this question, we studied dynamic properties at synapses to layer 2/3 pyramidal cells with intracellular recordings in slices of rat visual cortex. Synaptic responses were evoked by trains of test stimuli, which consisted of 10 pulses at different frequencies (5-40 Hz). Test stimulation was applied either without any adaptation (control) or 2 s after an adaptation stimulus, which consisted of 4 s stimulation of these same synapses at 10, 25, or 40 Hz. The synaptic parameters were then assessed from fitting the data with a model of synaptic dynamics. Our estimates of the synaptic parameters in control, without adaptation are broadly consistent with previous studies. Adaptation led to pronounced changes of synaptic transmission. After adaptation, the amplitude of the response to the first pulse in the test train decreased for several seconds and then recovered back to the control level with a time constant of 2-18 s. Analysis of the data with extended models, which include interaction between different pools of synaptic vesicles, suggests that the decrease of the response amplitude was due to a synergistic action of two factors, decrease of the release probability and depletion of the available transmitter. After a weak (10 Hz) adaptation, the decrease of the response amplitude was accompanied by and correlated with the decrease of the release probability. After a strong adaptation (25 or 40 Hz), the depletion of synaptic resources was the main cause for the reduced response amplitude. Adaptation also led to pronounced changes of the time constants of facilitation and recovery, however, these changes were not uniform in all synapses, and on the population level, the only consistent and significant effect was an acceleration of the recovery after a strong adaptation. Taken together, our results suggest, that apart from decreasing the amplitude of postsynaptic responses, adaptation may produce synapse-specific effects, which could result in a kind of re distribution of activity within neural networks. PMID- 15758050 TI - Androgen-induced vocal transformation in adult female African clawed frogs. AB - Sex-specific behaviors of some vertebrates are reversible by androgen administered in adulthood. Such behavioral transformations in adulthood provide opportunities to identify how neural systems reconfigure to produce sex-specific behavior. In this study, we focused on the vocalizations of the African clawed frog, Xenopus laevis. Male and female adult Xenopus produce sexually distinct vocalizations; males produce series of rapid clicks, whereas females produce slow trains of clicks. The differences in click rate can be reduced to differences in the firing rate of laryngeal motoneurons in vivo. This behavioral dimorphism is accompanied by various sex-specific characteristics throughout the vocal pathways, including functionally distinct laryngeal muscles and motoneurons in the sexes. In this study, we first determined whether and how testosterone (T) modifies the vocalizations of adult females and then examined changes underlying the behavioral modification at the laryngeal muscle and motoneuron levels. Our results show that, in response to T, the vocalizations of females were transformed within 13 wk. Vocal transformation was preceded by complete masculinization of muscle contractile properties and motoneuron soma size by the fourth week of T treatment, which suggests that the vocal pathways' peripheral components masculinize earlier than the behavior. Therefore the rate of transformation of vocal behavior must reflect a functional transformation of neurons in the central vocal pathways, which leads to the generation of male-like motor rhythms. PMID- 15758051 TI - Mapping of stimulus energy in primary visual cortex. AB - A recent optical imaging study of primary visual cortex (V1) by Basole, White, and Fitzpatrick demonstrated that maps of preferred orientation depend on the choice of stimuli used to measure them. These authors measured population responses expressed as a function of the optimal orientation of long drifting bars. They then varied bar length, direction, and speed and found that stimuli of a same orientation can elicit different population responses and stimuli with different orientation can elicit similar population responses. We asked whether these results can be explained from known properties of V1 receptive fields. We implemented an "energy model" where a receptive field integrates stimulus energy over a region of three-dimensional frequency space. The population of receptive fields defines a volume of visibility, which covers all orientations and a plausible range of spatial and temporal frequencies. This energy model correctly predicts the population response to bars of different length, direction, and speed and explains the observations made with optical imaging. The model also readily explains a related phenomenon, the appearance of motion streaks for fast moving dots. We conclude that the energy model can be applied to activation maps of V1 and predicts phenomena that may otherwise appear to be surprising. These results indicate that maps obtained with optical imaging reflect the layout of neurons selective for stimulus energy, not for isolated stimulus features such as orientation, direction, and speed. PMID- 15758052 TI - Cortical maps of separable tuning properties predict population responses to complex visual stimuli. AB - In the earliest cortical stages of visual processing, a scene is represented in different functional domains selective for specific features. Maps of orientation and spatial frequency preference have been described in the primary visual cortex using simple sinusoidal grating stimuli. However, recent imaging experiments suggest that the maps of these two spatial parameters are not sufficient to describe patterns of activity in different orientation domains generated in response to complex, moving stimuli. A model of cortical organization is presented in which cortical temporal frequency tuning is superimposed on the maps of orientation and spatial frequency tuning. The maps of these three tuning properties are sufficient to describe the activity in orientation domains that have been measured in response to drifting complex images. The model also makes specific predictions about how moving images are represented in different spatial frequency domains. These results suggest that the tangential organization of primary visual cortex can be described by a set of maps of separable neuronal receptive field features including maps of orientation, spatial frequency, and temporal frequency tuning properties. PMID- 15758053 TI - Changes in intrinsic properties of pyramidal neurons in adult rat S1 during cortical reorganization. AB - Peripheral denervation causes significant changes in the organization of developing or adult primary somatosensory cortex (S1). However, the basic mechanisms that underlie reorganization are not well understood. Most attention has been focused on possible synaptic mechanisms associated with reorganization. However, another important determinant of cortical circuit function is the intrinsic membrane properties of neurons in the circuit. Here we document changes in the intrinsic properties of pyramidal neurons in cortical layer 2/3 in adult rat primary somatosensory cortex (S1) after varying durations of forepaw denervation. Denervation of the forepaw induced a rapid and sustained shift in the location of the border between the forepaw and lower jaw representations of adult S1 (reorganization). Coronal slices from the reorganized region were maintained in vitro and the intrinsic properties of layer 2/3 pyramidal neurons of S1 were determined using whole cell recordings. In general, passive membrane properties were not affected by denervation; however, a variety of active properties were. The most robust changes were increases in the amplitudes of the fast and medium afterhyperpolarization (AHP) and an increase in the interval between action potentials (APs). Additional changes at some durations of denervation were observed for the AP threshold. These observations indicate that changes in intrinsic properties, mostly reflecting a decrease in overall excitation, may play a role in changes in cortical circuit properties during reorganization in adult S1, and suggest a possible role for AHPs in some of those changes. PMID- 15758054 TI - Regional and laminar differences in in vivo firing patterns of primate cortical neurons. AB - The firing rates of cortical neurons change in time; yet, some aspects of their in vivo firing characteristics remain unchanged and are specific to individual neurons. A recent study has shown that neurons in the monkey medial motor areas can be grouped into 2 firing types, "likely random" and "quasi-regular," according to a measure of local variation of interspike intervals. In the present study, we extended this analysis to area TE of the inferior temporal cortex and addressed whether this classification applies generally to different cortical areas and whether different types of neurons show different laminar distribution. We found that area TE did consist of 2 groups of neurons with different firing characteristics, one similar to the "likely random" type in the medial motor cortical areas, and the other exhibiting a "clumpy-bursty" firing pattern unique to TE. The quasi-regular type was rarely observed in area TE. The likely random firing type of neuron was more frequently found in layers V-VI than in layers II III, whereas the opposite was true for the clumpy-bursty firing type. These results show that neocortical areas consist of heterogeneous neurons that differ from one area to another in their basic firing characteristics. Moreover, we show that spike trains obtained from a single cortical neuron can provide a clue that helps to identify its layer localization. PMID- 15758055 TI - Visual-vestibular interactions during vestibular compensation: role of the pretectal not in horizontal VOR recovery after hemilabyrinthectomy in rhesus monkey. AB - Damage to the vestibular labyrinth leads to profound nystagmus and vertigo. Over time, the vestibular-ocular system recovers in a process called vestibular compensation leading to reduced nystagmus and vertigo provided visual signals are available. Our study was directed at identifying sources of visual information that could play a role in vestibular compensation. Specifically, we investigated the role of the pretectal nucleus of the optic tract (NOT) in vestibular compensation after hemilabyrinthectomy (HL) in rhesus monkeys. We chose the NOT because this structure provides critical visual motion information for adaptive modification of the vestibular ocular reflex (VOR). We produced bilateral NOT lesions by injecting the excitotoxin ibotenic acid. We compared vestibular compensation after HL in NOT-lesioned and control animals with intact NOTs. We measured eye movements with an electromagnetic method employing scleral search coils. Measurements included slow-phase eye velocity during spontaneous nystagmus, per- and postrotatory nystagmus and the horizontal VOR (hVOR) gain (eye-velocity/head velocity) associated with per- and postrotatory and sinusoidal (0.2-2.0 Hz; 30-90 degrees/s) whole body oscillation around the earth-vertical axis. VOR gain was low (<0.5) for rotation toward the HL side. Our control animal evinced significant vestibular compensation with VOR gains approaching unity by 100 days post HL. In contrast, monkeys with bilateral lesions of the NOT never obtained this significant recovery with hVOR gains well below unity at 100 days and beyond. Therefore our studies demonstrate that the NOT is an essential source of visual signals for the process of vestibular compensation after HL. PMID- 15758056 TI - Retinal bipolar cell input mechanisms in giant danio. III. ON-OFF bipolar cells and their color-opponent mechanisms. AB - Whole cell patch recording was performed from morphologically identified cone driven on-off bipolar cells (Cabs) in giant danio retinal slices to study their glutamate receptors and light-evoked responses. Specific agonists were puffed in the presence of cobalt, picrotoxin, and strychnine to identify glutamate receptors on these cells. Most Cabs responded to both the alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA)/kainate receptor agonist kainate and the excitatory amino acid transporter (EAAT) substrate D-aspartate, and both responses were localized to the dendrites. Kainate generated depolarizations whereas D-aspartate had E(rev) close to E(Cl) and generated hyperpolarizations, indicating that the AMPA/kainate receptors are sign-preserving, whereas the EAATs are sign-inverting. In response to white light, some Cabs gave on bipolar cell like responses whereas others gave off bipolar cell-like ones, but many cells' responses had both on and off bipolar cell components. In response to appropriately colored center-selective stimuli, many Cabs responded to short and long wavelengths with opposite polarities and were thus double color-opponent. The depolarizing components of the responses to white or colored stimuli were suppressed by the EAAT blocker DL-threo-beta-benzyloxyaspartate (TBOA), whereas the hyperpolarizing components were reduced by the AMPA/kainate receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX). These results are consistent with the hypothesis that both EAATs and AMPA/kainate receptors are involved in the generation of light-evoked responses in Cabs and that they confer these cells with on and off bipolar cell properties, respectively. Cabs can generate double color-opponent center responses by receiving inputs from certain cones through EAATs and from other cones through AMPA/kainate receptors. PMID- 15758057 TI - Mice lacking the major adult GABAA receptor subtype have normal number of synapses, but retain juvenile IPSC kinetics until adulthood. AB - There is a large variation in structurally and functionally different GABA(A) receptor subtypes. The expression pattern of GABA(A) receptor subunits is highly regulated, both temporarily and spatially. Especially during development, profound changes in subunit expression have been described. In most brain areas, the GABA(A) receptor alpha1 subunit replaces the alpha2 and/or alpha3 subunit as major alpha subunit. This is accompanied by a marked decrease in the open time of GABA(A) receptors and hence in the duration of postsynaptic responses. We describe here the development of GABAergic, synaptic transmission in mice lacking the alpha1 subunit. We show that alpha1 is to a large extent--but not entirely- responsible for the relatively short duration of postsynaptic responses in the developing and the mature brain. However, alpha1 already affects GABAergic transmission in the neonatal cerebral cortex when it is only sparsely expressed. It appears that the alpha1 -/- mice do not show a large reduction in GABAergic synapses but do retain long-lasting postsynaptic currents into adulthood. Hence, they form a good model to study the functional role of developmental GABA(A) receptor subunit switching. PMID- 15758064 TI - Long-term safety of bisphosphonates. PMID- 15758065 TI - What can we learn from gene expression profiling for adrenal tumor management? PMID- 15758066 TI - Frequent occurrence of hypogonadotropic hypogonadism in type 2 diabetes. PMID- 15758068 TI - Endocrine-related resources from the National Institutes of Health. PMID- 15758069 TI - The neurotrophin-3 receptor TrkC directly phosphorylates and activates the nucleotide exchange factor Dbs to enhance Schwann cell migration. AB - During the development of the peripheral nervous system, Schwann cells, the myelin-forming glia, migrate along axons before initiating myelination. We previously demonstrated that endogenous neurotrophin-3 (NT3) acting through the TrkC tyrosine kinase receptor enhances migration of premyelinating Schwann cells. This signaling pathway is mediated by the c-Jun N-terminal kinase (JNK) cascade regulated by the Rho GTPases Rac1 and Cdc42. However, missing is the link between TrkC and the GTPases. Here, we show that a guanine-nucleotide exchange factor (GEF), Dbl's big sister (Dbs), couples with TrkC to activate Cdc42 in Schwann cells. Furthermore, TrkC directly phosphorylates Dbs, thereby inducing the Cdc42 GEF activity. Taken together, activation of TrkC triggers Schwann cell migration by regulating Dbs upon direct tyrosine phosphorylation, providing a mechanism whereby a membrane receptor tyrosine kinase can induce the activation of Rho GTPase-GEFs. PMID- 15758070 TI - Biosynthesis of a D-amino acid in peptide linkage by an enzyme from frog skin secretions. AB - d-amino acids are present in some peptides from amphibian skin. These residues are derived from the corresponding L-amino acids present in the respective precursors. From skin secretions of Bombinae, we have isolated an enzyme that catalyzes the isomerization of an L-Ile in position 2 of a model peptide to D allo-Ile. In the course of this reaction, which proceeds without the addition of a cofactor, radioactivity from tritiated water is incorporated into the second position of the product. The amino acid sequence of this isomerase could be deduced from cloned cDNA and genomic DNA. After expression of this cDNA in oocytes of Xenopus laevis, isomerase activity could be detected. Polypeptides related to the frog skin enzyme are present in several vertebrate species, including humans. PMID- 15758071 TI - Long-term in vivo provision of antigen-specific T cell immunity by programming hematopoietic stem cells. AB - A method to genetically program mouse hematopoietic stem cells to develop into functional CD8 or CD4 T cells of defined specificity in vivo is described. For this purpose, a bicistronic retroviral vector was engineered that efficiently delivers genes for both alpha and beta chains of T cell receptor (TCR) to hematopoietic stem cells. When modified cell populations were used to reconstruct the hematopoietic lineages of recipient mice, significant percentages of antigen specific CD8 or CD4 T cells were observed. These cells expressed normal surface markers and responded to peptide antigen stimulation by proliferation and cytokine production. Moreover, they could mature into memory cells after peptide stimulation. Using TCRs specific for a model tumor antigen, we found that the recipient mice were able to partially resist a challenge with tumor cells carrying the antigen. By combining cells modified with CD8- and CD4-specific TCRs, and boosting with dendritic cells pulsed with cognate peptides, complete suppression of tumor could be achieved and even tumors that had become established would regress and be eliminated after dendritic cell/peptide immunization. This methodology of "instructive immunotherapy" could be developed for controlling the growth of human tumors and attacking established pathogens. PMID- 15758073 TI - Ultra-high vacuum scanning tunneling microscopy and theoretical studies of 1 halohexane monolayers on graphite. AB - A simple model system for the 2D self-assembly of functionalized organic molecules on surfaces was examined in a concerted experimental and theoretical effort. Monolayers of 1-halohexanes were formed through vapor deposition onto graphite surfaces in ultrahigh vacuum. Low-temperature scanning tunneling microscopy allowed the molecular conformation, orientation, and monolayer crystallographic parameters to be determined. Essentially identical noncommensurate monolayer structures were found for all 1-halohexanes, with differences in image contrast ascribed mainly to electronic factors. Energy minimizations and molecular dynamics simulations reproduced structural parameters of 1-bromohexane monolayers quantitatively. An analysis of interactions driving the self-assembly process revealed the crucial role played by small but anisotropic electrostatic forces associated with the halogen substituent. While alkyl chain dispersion interactions drive the formation of a close-packed adsorbate monolayer, electrostatic headgroup forces are found to compete successfully in the control of both the angle between lamella and backbone axes and the angle between surface and backbone planes. This competition is consistent with energetic tradeoffs apparent in adsorption energies measured in earlier temperature-programmed desorption studies. In accordance with the higher degree of disorder observed in scanning tunneling microscopy images of 1-fluorohexane, theoretical simulations show that electrostatic forces associated with the fluorine substituent are sufficiently strong to upset the delicate balance of interactions required for the formation of an ordered monolayer. The detailed dissection of the driving forces for self-assembly of these simple model systems is expected to aid in the understanding of the more complex self-assembly processes taking place in the presence of solvent. PMID- 15758072 TI - Reversible self-assembly and directed assembly of DNA-linked micrometer-sized colloids. AB - We present a technique for the directed assembly and self-assembly of micrometer scale structures based on the control of specific DNA linkages between colloidal particles. The use of DNA links combined with polymer brushes provides an effective way to regulate the range and magnitude of addressable forces between pairs (and further combinations) of different particles. We demonstrate that the autoassembly of alternate microbeads as well as their directed assembly, by using laser tweezers, is reversible. The key to reversibility is preventing the particles from falling into their van der Waals well at close distances. This goal is achieved by the use of adsorbed polymers that limit the number of DNA bridges to one to three between adjacent particles. PMID- 15758074 TI - High-dose radiation with bone marrow transfer prevents neurodegeneration in an inherited glaucoma. AB - Here, we show that high-dose gamma-irradiation accompanied with syngeneic bone marrow transfer can confer complete protection against glaucoma in a mouse model. Because bone marrow genotype was unaltered by this procedure, it was not the causative agent. The neuroprotection is robust and highly reproducible. Glaucoma prone DBA/2J mice received a single treatment at 5-8 weeks of age and were protected from glaucomatous retinal ganglion cell degeneration out to 14 months of age (oldest assessed). By 12-14 months, retinal ganglion cell degeneration is usually very severe and essentially complete in the majority of untreated DBA/2J mice. To assess reproducibility, three groups of mice were treated at different times, and the results were essentially the same each time. Considering all experiments, the vast majority of treated mice had no detectable glaucomatous neurodegeneration. A beneficial effect of treatment including high-dose radiation is unprecedented, and we are not aware of any other neuroprotective effects this substantial. Because of the robust protective effect, this treatment offers another tool for studying mechanisms of neuroprotection. PMID- 15758075 TI - Activation of pausing F1 motor by external force. AB - A rotary motor F(1), a catalytic part of ATP synthase, makes a 120 degrees step rotation driven by hydrolysis of one ATP, which consists of 80 degrees and 40 degrees substeps initiated by ATP binding and probably by ADP and/or P(i) dissociation, respectively. During active rotations, F(1) spontaneously fails in ADP release and pauses after a 80 degrees substep, which is called the ADP inhibited form. In the present work, we found that, when pushed >+40 degrees with magnetic tweezers, the pausing F(1) resumes its active rotation after releasing inhibitory ADP. The rate constant of the mechanical activation exponentially increased with the pushed angle, implying that F(1) weakens the affinity of its catalytic site for ADP as the angle goes forward. This finding explains not only its unidirectional nature of rotation, but also its physiological function in ATP synthesis; it would readily bind ADP from solution when rotated backward by an F(o) motor in the ATP synthase. Furthermore, the mechanical work for the forced rotation was efficiently converted into work for expelling ADP from the catalytic site, supporting the tight coupling between the rotation and catalytic event. PMID- 15758076 TI - Aminopeptidase N (CD13) functionally interacts with FcgammaRs in human monocytes. AB - Aminopeptidase N (E.C. 3.4.11.2) is a membrane-bound metalloproteinase expressed in many tissues. Although its cytoplasmic portion has only eight amino acids, cross-linking of CD13 by monoclonal antibodies (mAb) has been shown to trigger intracellular signaling. A functional association between CD13 and receptors for immunoglobulin G (FcgammaRs) has been proposed. In this work, we evaluated possible functional interactions between CD13 and FcgammaRs in human peripheral blood monocytes and in U-937 promonocytic cells. Our results show that during FcgammaR-mediated phagocytosis, CD13 redistributes to the phagocytic cup and is internalized into the phagosomes. Moreover, modified erythrocytes that interact with the monocytic cell membrane through FcgammaRI and CD13 are ingested simultaneously, more efficiently than those that interact through the FcgammaRI only. Also, co-cross-linking of CD13 with FcgammaRI by specific mAbs increases the level and duration of Syk phosphorylation induced by FcgammaRI cross-linking. Finally, FcgammaRI and CD13 colocalize in zones of cellular polarization and coredistribute after aggregation of either of them. These results demonstrate that CD13 and FcgammaRI can functionally interact on the monocytic cell membrane and suggest that CD13 may act as a signal regulator of FcgammaR function. PMID- 15758077 TI - The live vaccine strain of Francisella tularensis replicates in human and murine macrophages but induces only the human cells to secrete proinflammatory cytokines. AB - Francisella tularensis is the highly infectious agent of tularemia, a disease that can prove fatal in humans. An attenuated live vaccine strain (LVS) of this bacterium is avirulent in man but produces lethal illness in mice. As a step toward understanding the species specificity of the LVS, we compared its interactions with murine and human leukocytes. The bacterium replicated within murine bone marrow-derived macrophages (muBMDM), human monocyte-derived macrophages (huMDM), and freshly isolated human monocytes. However, the murine and human phagocytes differed in their ability to secrete proinflammatory cytokines in response to the LVS. The huMDM released large amounts of CXC chemokine ligand 8 (CXCL8) and CC chemokine ligand 2 when incubated with live or killed LVS organisms, and live bacteria also elicited production of interleukin 1beta (IL-1beta). Furthermore, human monocytes secreted CXCL8, IL-1beta, and tumor necrosis factor alpha in response to various bacterial preparations. In contrast, muBMDM produced little to no proinflammatory cytokines or chemokines when treated with any preparations of the LVS. Clearly, human and murine macrophages support growth of this bacterium. However, the greater proinflammatory response of human leukocytes to F. tularensis LVS may contribute to the avirulence of this strain in the human host. PMID- 15758078 TI - Complementary role of CD4+CD25+ regulatory T cells and TGF-beta in oral tolerance. AB - CD4(+)CD25(+) regulatory T cells are thought to be generated in the periphery as well as in the thymus. We sought to determine the roles played by CD4(+)CD25(+) T cells and transforming growth factor-beta (TGF-beta) in the induction and maintenance of tolerance generated by oral antigens in BALB/c mice. We found that oral administration of a high dose of ovalbumin (OVA) suppressed OVA-specific proliferation and antibody production in BALB/c mice depleted of CD25(+) cells. In contrast, the unresponsiveness induced by lower doses of OVA was only partially blocked by CD25 depletion prior to feeding. Depletion of CD4(+)CD25(+) cells after mice were orally tolerized did not reverse the tolerant status, indicating that these cells were not required to maintain the established tolerance. Furthermore, the induction of oral tolerance was not hampered by the administration of TGF-beta-neutralizing antibodies. However, in mice depleted of CD25(+) cells, anti-TGF-beta-neutralizing antibodies blocked the induction of tolerance, regardless of whether the mice followed the high- or low-dose regimens of oral OVA. CD25 depletion together with TGF-beta neutralization led the expansion of OVA-specific CD4 T cells against the subsequent antigen challenge, and each treatment alone did not. Our findings indicate that CD4(+)CD25(+) T cells and TGF-beta play a complementary role in the induction of oral tolerance, at least in part, by regulating the expansion of antigen-specific CD4 T cells. PMID- 15758079 TI - Orally administered OVA/CpG-ODN induces specific mucosal and systemic immune response in young and aged mice. AB - We have previously demonstrated that subcutaneously administered ovalbumin (OVA) plus synthetic oligodeoxynucleotides containing immunostimulatory CpG motifs (CpG ODN) as adjuvant stimulate cellular and humoral immunity and promote T helper cell type 1 differentiation in aged mice. The present study assessed the ability of CpG-ODN to induce an OVA-specific immune response after oral immunization in young (3-month-old) and aged (18-month-old) BALB/c mice. Oral OVA/CpG-ODN immunization induces a similar OVA-specific T cell-proliferative response (in mucosal and systemic tissues), immunoglobulin G (IgG) in plasma, and IgA in intestinal washes in both groups of ages. The OVA-specific humoral immune response observed in aged mice was similar to the one observed in young mice, peaking at day 7 after the last oral immunization and was present over 40 days after the last oral immunization. The pattern of cytokines released in culture supernatants in both groups of mice was similar, with specific interferon-gamma secretion in the absence of interleukin-5 responses. These results provide evidence that orally administered OVA/CpG-ODN induces a young-like, specific, immune response against OVA in aged mice, showing that CpG-ODN might be used as a mucosal adjuvant during aging. PMID- 15758080 TI - Confidential enquiries into maternal deaths: 50 years of closing the loop. PMID- 15758081 TI - Maternal deaths from anaesthesia. An extract from Why Mothers Die 2000-2002, the Confidential Enquiries into Maternal Deaths in the United Kingdom: Chapter 9: Anaesthesia. AB - This is the first of two extracts from Why Mothers Die 2000-2002, issued on 12 November 2004 by the Confidential enquiry into Maternal and Child Health (CEMACH), reproduced with permission. The full report can be accessed via their web site: http://www.cemach.org.uk/ PMID- 15758082 TI - Maternal deaths from anaesthesia. An extract from Why Mothers Die 2000-2002, the Confidential Enquiries into Maternal Deaths in the United Kingdom: Chapter 17: Trends in intensive care. AB - This is the second of two extracts from Why Mothers Die 2000-2002, issued on 12 November 2004 by the Confidential enquiry into Maternal and Child Health (CEMACH), reproduced with permission. The full report can be accessed via their web site: http://www.cemach.org.uk/ PMID- 15758083 TI - Postpartum post-dural puncture headache. PMID- 15758084 TI - Reduction in mortality from severe head injury following introduction of a protocol for intensive care management. PMID- 15758085 TI - ILMA in cervical spine immobilization. PMID- 15758086 TI - Levobupivacaine base and levobupivacaine hydrochloride. PMID- 15758087 TI - Aspiration and the laryngeal mask airway. PMID- 15758088 TI - Anaphylaxis and anaesthesia. PMID- 15758089 TI - CobraPLA as a conduit for flexible bronchoscopy in a child under general anaesthesia. PMID- 15758090 TI - Comments on "Improvements to measuring water flux in the vadose zone" (K.C. Masarik, J.M. Norman, K.R. Brye, and J.M. Baker; J. Environ. Qual. 33:1152-1158). PMID- 15758092 TI - Crusting of stored dairy slurry to abate ammonia emissions: pilot-scale studies. AB - Storage of cattle slurry is a significant source of ammonia (NH3) emissions. Emissions can be reduced by covering slurry stores, but this can incur significant costs, as well as practical and technical difficulties. In this pilot scale study, slurry was stored in small tanks (500 L) and the effectiveness of natural crust development for reducing NH3 emissions was assessed in a series of experiments. Also, factors important in crust development were investigated. Measurements were made of crust thickness and specially adapted tank lids were used to measure NH3 emissions. Slurry dry matter (DM) content was the most important factor influencing crust formation, with no crust formation on slurries with a DM content of <1%. Generally, crusts began to form within the first 10 to 20 d of storage, at which time NH3 emission rates would decrease. The formation of a natural crust reduced NH3 emissions by approximately 50%. The type of bedding used in the free stall barn did not influence crust formation, nor did ambient temperature or air-flow rate across the slurry surface. There was a large difference in crust formation between slurries from cattle fed a corn (Zea mays L.) silage-based diet and those fed a grass silage-based diet, although dietary differences were confounded with bedding differences. The inclusion of a corn starch and glucose additive promoted crust formation and reduced NH3 emission. The maintenance of a manageable crust on cattle slurry stores is recommended as a cost-effective means of abating NH3 emissions from this phase of slurry management. PMID- 15758093 TI - Empirical relationship between use, area, and ambient air concentration of methyl bromide. AB - Methyl bromide (MeBr) is one of the most widely used soil fumigants. Human exposure to MeBr above threshold values can cause serious health problems. The exposure assessment of MeBr depends on estimation or measurement of its air concentrations. This study proposed a methodology for systematically exploring the empirical relationship between MeBr use intensity and ambient air concentrations. Monitored air concentrations were regressed to MeBr use over various spatiotemporal scales that step-wise increased around the monitoring site and monitoring period. The results showed that the goodness-of-fit varied with the spatiotemporal scale of MeBr use. The best fit was Y = 0.46 + 0.00120X (R2 = 0.95, n = 11), where Y was the 8-wk average ambient air concentration (microg/m3), and X was the weekly average use (kg/wk) over an area of 11.3 x 11.3 km (7 x 7 mi). The model was calibrated with air-monitoring data and use data of 2000, and verified with the same type data of 2001. The model estimated subchronic air concentration with reasonable accuracy. PMID- 15758094 TI - The relationship between greenhouse gas emissions and the intensity of milk production in Ireland. AB - European Union agri-environmental schemes aim to reduce the environmental impact of agricultural production, but were developed before consideration of greenhouse gas emissions from agriculture. Life cycle assessment methodology provided a framework for comparing emissions as kg CO2 equivalent per kg of energy corrected milk (ECM) (kg CO2 kg(-1) ECM yr(-1)) and per hectare (kg CO2 ha(-1) yr(-1)) for farms both within and outside the Irish agri-environmental scheme. The agri environmental scheme farms operate extensive systems from 40 to 120 cows producing between 3032 and 5946 kg ECM cow(-1) lactation(-1). The cows are fed on grass, conserved silage, and concentrates. Supplementation ranged between 250 and 620 kg cow(-1) yr(-1). The conventional farms had between 30 and 77 milking cows producing 4736 to 6944 kg ECM cow(-1) lactation(-1). Supplementation ranged from 400 to 1000 kg cow(-1) yr(-1). The emissions from each unit were estimated using published emissions factors and possible error was evaluated by using ranges for each factor. Calculated emissions ranged from 0.92 to 1.51 kg CO2 kg(-1) ECM yr( 1) and 5924 to 8323 kg CO2 ha(-1). On average, total emissions from conventional farms were around 18% (p = 0.01) greater than the agri-environmental scheme farms and emissions per hectare (total area required) were 17% greater (p = 0.02) but there was no significant difference (p = 0.335) in terms of emission per unit milk produced. To evaluate greenhouse gas emissions for each farm in terms of the system intensity it was necessary to define a measure of intensification and area per liter of milk produced that was best. PMID- 15758095 TI - Tillage and crop residue effects on soil carbon and carbon dioxide emission in corn-soybean rotations. AB - Soil C change and CO2 emission due to different tillage systems need to be evaluated to encourage the adoption of conservation practices to sustain soil productivity and protect the environment. We hypothesize that soil C storage and CO2 emission respond to conservation tillage differently from conventional tillage because of their differential effects on soil properties. This study was conducted from 1998 through 2001 to evaluate tillage effects on soil C storage and CO2 emission in Clarion-Nicollet-Webster soil association in a corn [Zea mays L.]-soybean [Glycine max (L.) Merr.] rotation in Iowa. Treatments included no tillage with and without residue, strip-tillage, deep rip, chisel plow, and moldboard plow. No-tillage with residue and strip-tillage significantly increased total soil organic C (TC) and mineral fraction C (MFC) at the 0- to 5- and 5- to 10-cm soil depths compared with chisel plow after 3 yr of tillage practices. Soil CO2 emission was lower for less intensive tillage treatments compared with moldboard plow, with the greatest differences occurring immediately after tillage operations. Cumulative soil CO2 emission was 19 to 41% lower for less intensive tillage treatments than moldboard plow, and it was 24% less for no-tillage with residue than without residue during the 480-h measurement period. Estimated soil mineralizable C pool was reduced by 22 to 66% with less intensive tillage treatments compared with moldboard plow. Adopting less intensive tillage systems such as no-tillage, strip-tillage, deep rip, and chisel plow and better crop residue cover are effective in reducing CO2 emission and thus improving soil C sequestration in a corn-soybean rotation. PMID- 15758096 TI - Assessment of denitrification gaseous end-products in the soil profile under two water table management practices using repeated measures analysis. AB - The denitrification process and nitrous oxide (N2O) production in the soil profile are poorly documented because most research into denitrification has concentrated on the upper soil layer (0-0.15 m). This study, undertaken during the 1999 and 2000 growing seasons, was designed to examine the effects of water table management (WTM), nitrogen (N) application rate, and depth (0.15, 0.30, and 0.45 m) on soil denitrification end-products (N2O and N2) from a corn (Zea mays L.) field. Water table management treatments were free drainage (FD) with open drains and subirrigation (SI) with a target water table depth of 0.6 m. Fertility treatments (ammonium nitrate) were 120 kg N ha(-1) (N120) and 200 kg N ha(-1) (N200). During both growing seasons greater denitrification rates were measured in SI than in FD, particularly in the surface soil (0-0.15 m) and at the intermediate (0.15-0.30 m) soil depths under N200 treatment. Greater denitrification rates under the SI treatment, however, were not accompanied with greater N2O production. The decrease in N2O production under SI was probably caused by a more complete reduction of N2O to N2, which resulted in lower N2O to (N2O + N2) ratios. Denitrification rate, N2O production and N2O to (N2O + N2) ratios were only minimally affected by N treatments, irrespective of sampling date and soil depth. Overall, half of the denitrification occurred at the 0.15- to 0.30- and 0.30- to 0.45-m soil layers, and under SI, regardless of fertility treatment level. Consequently, sampling of the 0- to 0.15-m soil layer alone may not give an accurate estimation of denitrification losses under SI practice. PMID- 15758097 TI - Methane oxidation in slurry storage surface crusts. AB - Livestock manure is a significant source of atmospheric methane (CH4), especially during liquid storage. In liquid manure (slurry) storages a surface crust may form naturally, or an artificial surface crust can be established. We investigated whether there is a potential for CH4 oxidation in this environment. Surface crust materials were sampled from experimental storages with cattle slurry (with natural crust) or anaerobically digested cattle slurry (with straw layer) that had been stored with or without a wooden cover. Extracts of surface crust material were incubated with 5.6% CH4 in the headspace, and methanotrophic activity was demonstrated in all four treatments following a 4- to 10-d lag phase. Subsequent incubation of field-moist surface crust material with 350 microL L(-1) CH4 also showed CH4 oxidation, indicating a potential for CH4 removal under practical storage conditions. There was no CH4 oxidation activity during incubation of autoclaved samples. Methane oxidation rates were 0.1 to 0.5 mg kg(-1) organic matter (OM) h(-1), which is comparable with the activity in wetlands and rice paddies. Partial drying increased CH4 oxidation to 0.2 to 1.4 mg kg(-1) OM h(-1), probably as a result of improved diffusivity within the surface crust. Rewetting reversed the stimulation of methanotrophic activity in some treatments, but not in others, possibly due to a decline in CH4 production in anaerobic volumes, or to growth of methanotrophs during incubation. This study presents direct evidence for methanotrophic activity in slurry storages. Measures to ensure crust formation with or without a solid cover appear to be a cost effective greenhouse gas mitigation option. PMID- 15758098 TI - 9,10-Phenanthrenequinone photoautocatalyzes its formation from phenanthrene, and inhibits biodegradation of naphthalene. AB - Polycyclic aromatic hydrocarbons (PAHs) have earned considerable attention due to their widespread environmental distribution and toxicity. In the environment, PAHs decompose by a variety of biotic and abiotic pathways. In both polar and nonpolar environments, phenanthrene (Phe, a common, three-ring PAH) is converted by sunlight to more polar products such as 9,10-phenanthrenequinone (PheQ) and subsequent oxidation products such as the corresponding open-ring dicarboxylic acid product. Biodegradation of phenanthrene also usually leads to oxidative metabolites, and eventually ends in mineralization. Our experimental objective was to investigate the photodegradation of phenanthrene and determine the effect of reaction products such as PheQ on microbial biodegradation of two- and three ring PAHs. Abiotic experiments were performed to examine the photolytic breakdown of Phe; Phe was converted to PheQ, which catalyzed its own formation. In biodegradation experiments PheQ (0.04-4 mg/L) caused marked inhibition of naphthalene (Nap) biodegradation by a Burkholderia species; Phe did not. Only 20% of the naphthalene was degraded in the presence of PheQ compared with 75% in the control culture with no PheQ added. No PAH-degrading cultures were able to use PheQ as sole carbon source; however, the Phe-degrading enrichment culture dominated by a Sphingomonas species was able to degrade PheQ cometabolically in the presence of Phe. These results may explain why photooxidized phenanthrene containing mixtures can resist biodegradation. PMID- 15758099 TI - Straining, attachment, and detachment of cryptosporidium oocysts in saturated porous media. AB - Accurate knowledge of the transport and deposition behavior for pathogenic Cryptosporidium parvum oocysts is needed to assess contamination and protect water resources. Experimental and modeling studies were undertaken to examine the roles of attachment, detachment, and straining on oocyst transport and retention. Saturated column studies were conducted using Ottawa aquifer sands (U.S. Silica, Ottawa, IL) with median grain sizes of 710, 360, and 150 microm. Decreasing the median sand size tended to produce lower effluent concentrations, greater oocyst retention in the sand near the column inlet, and breakthrough of oocysts at later times. Oocyst transport data also exhibited concentration tailing. Mathematical modeling of the oocyst transport data using fitted first-order attachment and detachment coefficients provided a satisfactory description of the observed effluent concentration curves, but a poor characterization of the oocyst spatial distribution. Modeling of these data using an irreversible straining term that is depth dependent provided a better description of the oocyst spatial distribution, but could not account for the observed effluent concentration tailing or late breakthrough times. A more physically realistic description of the data was obtained by modeling attachment, detachment, and straining. The percentage of total oocysts retained by straining was estimated from effluent mass balance considerations to be 68% for 710-microm sand, 79% for 360-microm sand, and 87% for 150-microm sand. Straining coefficients were then selected to achieve these percentages of total oocyst retention, and attachment and detachment coefficients were fitted to the effluent concentration curves. Dramatic differences in the predicted oocyst breakthrough curves were observed at greater transport distances for the various model formulations (inclusion or exclusion of straining). Justification for oocyst straining was provided by trends in the transport data, simulation results, pore size distribution information, and published literature. PMID- 15758100 TI - Desorption kinetics of arsenate from kaolinite as influenced by pH. AB - Arsenic is highly toxic and therefore represents a potential threat to the environment and human health. The mobility and bioavailability of arsenic in soil is mostly controlled by adsorption and desorption reactions. Even though adsorption and traditional batch desorption experiments provide information about the environmental fate of As, the equilibrium conditions imposed in these studies would usually not be reached in the natural environment. Flow-through desorption techniques, where the desorbed species are removed from the substrate, can therefore be used to provide information about the rate and mechanisms of As desorption. The effect of pH on As adsorption reactions is relatively well understood; however, desorption of As and the effect of pH on As desorption remain unexplored. Desorption of As(V) (the most dominant arsenic species in aerated soils) was therefore investigated using batch and flow-through desorption experiments. Traditional batch desorption experiments underestimated the desorption rate of As(V) from kaolinite. The pH had a large effect on the amount of As(V) desorbed from kaolinite, with both an increase and a decrease in pH (from the initial pH 6.4) enhancing As(V) desorption. Modeling desorption over time revealed that the pH can influence As(V) desorption over extended periods of time. PMID- 15758101 TI - Removal of selenate from water by zerovalent iron. AB - Zerovalent iron (ZVI) has been widely used in the removal of environmental contaminants from water. In this study, ZVI was used to remove selenate [Se(VI)] at a level of 1000 microg L(-1) in the presence of varying concentrations of Cl-, SO(2-)4, NO(-)3, HCO(-)3, and PO(3-)4. Results showed that Se(VI) was rapidly removed during the corrosion of ZVI to iron oxyhydroxides (Fe(OH)). During the 16 h of the experiments, 100 and 56% of the added Se(VI) was removed in 10 mM Cl- and SO(2-)4 solutions under a closed contained system, respectively. Under an open condition, 100 and 93% of the added Se(VI) were removed in the Cl- and SO(2 )4 solutions, respectively. Analysis of Se species in ZVI-Fe(OH) revealed that selenite [Se(IV)] and nonextractable Se increased during the first 2 to 4 h of reaction, with a decrease of Se(VI) in the Cl- experiment and no detection of Se(VI) in the SO(2-)4 experiment. Two mechanisms can be attributed to the rapid removal of Se(VI) from the solutions. One is the reduction of Se(VI) to Se(IV), followed by rapid adsorption of Se(IV) to Fe(OH). The other is the adsorption of Se(VI) directly to Fe(OH), followed by its reduction to Se(IV). The results also show that there was little effect on Se(VI) removal in the presence of Cl- (5, 50, and 100 mM), NO(-)3 (1, 5, and 10 mM), SO(2-)4 (5 mM), HCO(-)3 (1 and 5 mM), or PO(3-)4 (1 mM) and only a slight effect in the presence of SO(2-)4 (50 and 100 mM), HCO(-)3 (10 mM), and PO(3-)4 (5 mM) during a 2-d experiment, whereas 10 mM PO(3-)4 significantly inhibited Se(VI) removal. This work suggests that ZVI may be an effective agent to remove Se from Se-contaminated agricultural drainage water. PMID- 15758102 TI - Prediction of zinc, cadmium, lead, and copper availability to wheat in contaminated soils using chemical speciation, diffusive gradients in thin films, extraction, and isotopic dilution techniques. AB - To predict the availability of metals to plants, it is important to understand both solution- and solid-phase processes in the soil, including the kinetics of metal release from its binding agent (ligand and/or particle). The present study examined the speciation and availability of Zn, Cd, Pb, and Cu in a range of well equilibrated metal-contaminated soils from diverse sources using several techniques as a basis for predicting metal uptake by plants. Wheat (Triticum aestivum L.) was grown in 13 metal-contaminated soils and metal tissue concentrations (Zn, Cd, Pb, and Cu) in plant shoots were compared with total soil metal concentrations, total soluble metal, and free metal activities (pM2+) in soil pore waters, 0.01 M CaCl2-extractable metal concentrations, E values measured by isotope dilution, and effective metal concentrations, C(E), measured by diffusive gradients in thin films (DGT). In the DGT technique, ions are dynamically removed by their diffusion through a gel to a binding resin, while E values represent the isotopically exchangeable (labile) metal pools. Free metal activities (Zn2+, Cd2+, and Pb2+) in soil pore waters were determined using a Donnan dialysis technique. Plant Zn and Cd concentrations were highly related to C(E), while relationships for Zn and Cd with respect to the other measures of metals in the soils were generally lower, except for CaCl2-extractable Cd. These results suggest that the kinetically labile solid-phase pool of metal, which is included in the DGT measurement, played an important role in Zn and Cd uptake by wheat along with the labile metal in soil solution. Plant Pb concentrations were highly related to both soil pore water concentrations and C(E), indicating that supply from the solid phase may not be so important for Pb. Predictions of Cu uptake by wheat from these soils by the various measures of Cu were generally poor, except surprisingly for total Cu. PMID- 15758103 TI - Copper availability in seven Israeli soils incubated with and without biosolids. AB - Land application is becoming a preferred option for disposal of sewage sludge (biosolids) from wastewater treatment plants. However, it creates potential risks due to the heavy metal contents of these materials, with copper (Cu) being of chief concern. The long-term fate of biosolid metals applied to agricultural soils is not well understood, particularly in the soils of the Middle East. This investigation was conducted to determine whether the availability of Cu changes with time in biosolid-amended and nonamended soils from Israel. Seven soils, typifying the span of properties and formation environments encountered in Israel, were incubated with and without biosolids for 7 yr, and changes in organic carbon (OC) content and labile Cu concentration were determined. Isotopic exchange techniques, using 64Cu, and ion activity measurements, using a Cu2+ ion selective electrode, revealed that the available Cu concentration remained relatively low and stable over the 7-yr incubation. This was despite substantial reductions in OC. This study shows that, with regard to Cu, application of such biosolids to these soils at rates of up to 250 Mg ha(-1) does not pose a threat to the environment in the short to medium term. PMID- 15758104 TI - Characterizing land surface erosion from cesium-137 profiles in lake and reservoir sediments. AB - Recognition of the threat to the sustainable use of the earth's resources posed by soil erosion and associated off-site sedimentation has generated an increasing need for reliable information on global rates of soil loss. Existing methods of assessing rates of soil loss across large areas possess many limitations and there is a need to explore alternative approaches to characterizing land surface erosion at the regional and global scale. The downcore profiles of 137Cs activity available for numerous lakes and reservoirs located in different areas of the world can be used to provide information on land surface erosion within the upstream catchments. The rate of decline of 137Cs activity toward the surface of the sediment deposited in a lake or reservoir can be used to estimate the rate of surface lowering associated with eroding areas within the upstream catchment, and the concentration of 137Cs in recently deposited sediment provides a basis for estimating the relative importance of surface and channel, gully, and/or subsurface erosion as a source of the deposited sediment. The approach has been tested using 137Cs data from several lakes and reservoirs in southern England and China, spanning a wide range of specific suspended sediment yield. The results obtained are consistent with other independent evidence of erosion rates and sediment sources within the lake and reservoir catchments and confirm the validity of the overall approach. The approach appears to offer valuable potential for characterizing land surface erosion, particularly in terms of its ability to provide information on the rate of surface lowering associated with the eroding areas, rather than an average rate of lowering for the entire catchment surface. PMID- 15758105 TI - In situ ground water denitrification in stratified, permeable soils underlying riparian wetlands. AB - The ground water denitrification capacity of riparian zones in deep soils, where substantial ground water can flow through low-gradient stratified sediments, may affect watershed nitrogen export. We hypothesized that the vertical pattern of ground water denitrification in riparian hydric soils varies with geomorphic setting and follows expected subsurface carbon distribution (i.e., abrupt decline with depth in glacial outwash vs. negligible decline with depth in alluvium). We measured in situ ground water denitrification rates at three depths (65, 150, and 300 cm) within hydric soils at four riparian sites (two per setting) using a 15N enriched nitrate "push-pull" method. No significant difference was found in the pattern and magnitude of denitrification when grouping sites by setting. At three sites there was no significant difference in denitrification among depths. Correlations of site characteristics with denitrification varied with depth. At 65 cm, ground water denitrification correlated with variables associated with the surface ecosystem (temperature, dissolved organic carbon). At deeper depths, rates were significantly higher closer to the stream where the subsoil often contains organically enriched deposits that indicate fluvial geomorphic processes. Mean rates ranged from 30 to 120 microg N kg(-1) d(-1) within 10 m versus <1 to 40 microg N kg(-1) d(-1) at >30 m from the stream. High denitrification rates observed in hydric soils, down to 3 m within 10 m of the stream in both alluvial and glacial outwash settings, argue for the importance of both settings in evaluating the significance of riparian wetlands in catchment scale N dynamics. PMID- 15758106 TI - Assessment of herbicide leaching risk in two tropical soils of Reunion Island (France). AB - Application of organic chemicals to a newly irrigated sugarcane (Saccharum officinarum L.) area located in the semiarid western part of Reunion Island has prompted local regulatory agencies to determine their potential to contaminate ground water resources. For that purpose, simple indices known as the ground water ubiquity score (Gustafson index, GUS), the retardation factor (RF), the attenuation factor (AF), and the log-transformed attenuation factor (AFT) were employed to assess the potential leaching of five herbicides in two soil types. The herbicides were alachlor [2-chloro-2',6'-diethyl-N-(methoxy-methy) acetanilide], atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-1,3,5 triazine], diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea], 2,4-D [(2,4 dichlorophenoxy) acetic-acid], and triclopyr [((3,5,6-trichloro-2-pyridyl)oxy) acetic-acid]. The soil types were Vertic (BV) and Andepts (BA) Inceptisols, which are present throughout the Saint-Gilles study area on Reunion Island. To calculate the indices, herbicide sorption (K(oc)) and dissipation (half-life, DT50) properties were determined from controlled batch experiments. Water fluxes below the root zone were estimated by a capacity-based model driven by a rainfall frequency analysis performed on a 13-yr data series. The results show a lower risk of herbicide leaching than in temperate regions due to the tropical conditions of the study area. Higher temperatures and the presence of highly adsorbent soils may explain smaller DT50 and higher K(oc) values than those reported in literature concerning temperate environments. Based on the RF values, only 2,4-D and triclopyr appear mobile in the BV soil, with all the other herbicides being classified from moderately to very immobile in both soils. The AFT values indicate that the potential leaching of the five herbicides can be considered as unlikely, except during the cyclonic period (about 40 d/yr) when there is a 2.5% probability of recharge rates equal to or higher than 50 mm/d. In that case, atrazine in both soils, 2,4-D and triclopyr in the BV soil, and diuron and alachlor in the BA soil present a high risk of potential contamination of ground water resources. PMID- 15758107 TI - Characterization of the pesticide chlordane in estuarine river sediments. AB - Sediments are increasingly recognized as both carrier and potential source of contaminants in aquatic environments. This study investigated the characteristics and spatial distribution of total chlordane and its three most abundant compounds, including alpha-chlordane, gamma-chlordane, and trans-nonachlor, in sediments from the Cedar and Ortega rivers, Florida, USA, using geographic information system (GIS)-based kriging analyses and field measurements. Kriging analysis showed that two areas, one from the Cedar River area and the other from the northern end of the Ortega River area, were contaminated. The maximum concentrations of total chlordane, gamma-chlordane, alpha-chlordane, and trans nonachlor in the sediments were, respectively, 101.8, 20.1, 26.3, and 19.2 microg/kg. A plot of total organic carbon (TOC)-normalized chlordane concentrations showed that effects of grain size on sediment chlordane contamination were negligible. A principal axis analysis further revealed that a linear correlation existed between alpha-chlordane and total chlordane as well as between gamma-chlordane and total chlordane, whereas no correlation existed between trans-nonachlor and total chlordane. Comparison of total chlordane concentration with Florida Sediment Assessment Guidelines showed that the Cedar River and the northern end of the Ortega River had total chlordane concentrations above the probable effect level (4.79 microg/kg), which could pose a potential risk to aquatic life. PMID- 15758109 TI - Influence of phytase on water-soluble phosphorus in poultry and swine manure. AB - The effect of dietary non-phytin phosphorus (NPP) and phytase (PHY) concentration on total phosphorus (TP) and water-soluble phosphorus (WSP) excretion was determined. Diets tested in broiler experiments were: National Research Council nutrient requirements for non-phytin phosphorus (NRC), NRC + PHY, reduced non phytin phosphorus (RED), and RED + PHY. Turkey and swine experiment diets included NRC, RED, and RED + PHY. For all experiments, except broiler Experiment 1, excreta were: (i) boiled, antibiotic added, then frozen; (ii) boiled, antibiotic added, incubated (37 degrees C for 72 h), then frozen; and (iii) incubated, boiled, antibiotic added, then frozen. In Experiment 1, excreta were collected and frozen or incubated for 24 or 48 h. In broiler Experiment 1, WSP was not affected by phytase but increased with post-excretion incubation. In a broiler Experiment 2, reducing NPP resulted in reduced excreta TP and WSP (11.3 to 8.3 and 5.3 to 2.7 g kg(-1)). Feeding RED + PHY diets resulted in less TP and WSP (7.6 and 0.6 g kg(-1)) as compared with NRC + PHY (11.2 and 3.9 g kg(-1), Experiment 3). Incubation resulted in increased WSP, irrespective of phytase addition such that WSP as a percent of TP was similar among treatments. Addition of antibiotics before incubation prevented the increase in WSP. Similar results were observed with turkey and swine. Therefore, when phytase is used properly (i.e., with a simultaneous reduction of NPP), WSP or WSP as a percent of TP are not affected. The increase in WSP as a percent of TP post-excretion is a function of excreta microbial activity and not dietary phytase addition. PMID- 15758108 TI - Interactions of organic compounds with wastewater dissolved organic matter: role of hydrophobic fractions. AB - The role of structural fractions of dissolved organic matter (DOM) from wastewater in the sorption process of hydrophobic organic compounds is still not clear. In this study, DOM from two wastewater treatment plants (Lachish and Netanya, Israel) was fractionated to hydrophobic acid (HoA) and hydrophobic neutral (HoN) fractions. The fractions were characterized and their sorptive capabilities for s-triazine herbicides and polycyclic aromatic hydrocarbons (PAHs) were studied. For all sorbates, the binding to the HoN fractions was much higher than to HoA fractions. The HoA fractions were more polar than the HoN fractions, containing a higher level of carboxylic functionalities. However the higher binding coefficients of atrazine (2-chloro-4-ethylamine-6-isopropylamino-s triazine) and ametryn [2-(ethylamino)-4-isopropylamino-6-methyl-thio-s-triazine] obtained for the HoN fractions suggest that their sorption is governed by hydrophobic-like interactions rather than H bonding. The values of binding coefficients of PAHs measured for the HoN fractions were within the range reported for humic acids and much higher than other fractions, suggesting that this fraction plays an important role in the overall sorption of these compounds by DOM. Higher sorption coefficients were measured for the Netanya DOM sample containing higher level of hydrophobic fractions (HoA + HoN) than the Lachish DOM, suggesting that the sorption of hydrophobic organic compounds by DOM is governed by the level of these structural substances. The evaluation of mobility of organic pollutants by wastewater irrigation requires not only assessment of the total carbon concentration but also, more importantly, the content of the hydrophobic fractions. PMID- 15758110 TI - Relating soil phosphorus to dissolved phosphorus in runoff: a single extraction coefficient for water quality modeling. AB - Phosphorus transport from agricultural soils contributes to eutrophication of fresh waters. Computer modeling can help identify agricultural areas with high potential P transport. Most models use a constant extraction coefficient (i.e., the slope of the linear regression between filterable reactive phosphorus [FRP] in runoff and soil P) to predict dissolved P release from soil to runoff, yet it is unclear how variations in soil properties, management practices, or hydrology affect extraction coefficients. We investigated published data from 17 studies that determined extraction coefficients using Mehlich-3 or Bray-1 soil P (mg kg( 1)), water-extractable soil P (mg kg(-1)), or soil P sorption saturation (%) as determined by ammonium oxalate extraction. Studies represented 31 soils with a variety of management conditions. Extraction coefficients from Mehlich-3 or Bray 1 soil P were not significantly different for 26 of 31 soils, with values ranging from 1.2 to 3.0. Extraction coefficients from water-extractable soil P were not significantly different for 17 of 20 soils, with values ranging from 6.0 to 18.3. The relationship between soil P sorption saturation and runoff FRP (microg L(-1)) was the same for all 10 soils investigated, exhibiting a split-line relationship where runoff FRP rapidly increased at P sorption saturation values greater than 12.5%. Overall, a single extraction coefficient (2.0 for Mehlich-3 P data, 11.2 for water-extractable P data, and a split-line relationship for P sorption saturation data) could be used in water quality models to approximate dissolved P release from soil to runoff for the majority of soil, hydrologic, or management conditions. A test for soil P sorption saturation may provide the most universal approximation, but only for noncalcareous soils. PMID- 15758111 TI - Resuspension of sediment-associated Escherichia coli in a natural stream. AB - In this study, a tracer bacteria was used to investigate the resuspension and persistence of sediment-associated bacteria in a small alluvial stream. The study was conducted in Swan Creek, located within the Grand River watershed of Ontario, Canada. A 1.1-m2 section of the bed was seeded with a strain of Escherichia coli resistant to nalidixic acid (E. coli NAR). The survival, transport, and redistribution of the tracer bacteria within a 1.7-km river section downstream of the source cell was assessed for a 2-mo period following the introduction of the tracer bacteria. This study has illustrated that enteric bacteria can survive in bed sediments for up 6 wk and that inactivation of the tracer bacteria resembled typical first-order decay. Critical conditions for resuspension, as well as resuspension rates, of sediment-associated bacteria were determined for several storm events. The critical shear stress for E. coli NAR resuspension in Swan Creek ranged from 1.5 to 1.7 N m(-2), which is comparable with literature values for critical shear stresses for erosion of cohesive sediments. Bacteria resuspension was primarily limited to the rising limb of storm hydrographs implying that a finite supply of sediment-associated bacteria are available for resuspension during individual storm events. The information presented in this paper will further the development of representative microbial water quality models. PMID- 15758112 TI - Nitrate losses in subsurface drainage from a corn-soybean rotation as affected by fall and spring application of nitrogen and nitrapyrin. AB - Substantial amounts of NO3 from agricultural crop production systems on poorly drained soils can be transported to surface water via subsurface drainage. A field study was conducted from the fall of 1993 through 2000 on a tile-drained Canisteo clay loam soil (fine-loamy, mixed, superactive, calcareous, mesic Typic Endoaquoll) to determine the influence of fall vs. spring application of N and nitrapyrin [NP; 2-chloro-6-(trichloromethyl) pyridine] on NO3 losses from a corn (Zea mays L.)-soybean [Glycine max (L.) Merr.] rotation. Four anhydrous ammonia treatments (fall N, fall N + NP, spring preplant N, and spring N + NP) were replicated four times and applied at 135 kg N ha(-1) for corn on individual drainage plots. Drainage occurred in all seven years. Seventy-one percent of the annual drainage and 75% of the annual NO3 loss occurred in April, May, and June. Fifty-four percent of the NO3 lost in the drainage occurred during the corn phase and 46% during the soybean phase. Annual flow-weighted NO3-N concentrations for the fall, fall + NP, spring, and spring + NP treatments averaged 14.3, 11.5, 10.7, and 11.3 mg L(-1) during the corn phase but annual NO3-N concentrations were still > or =10 mg L(-1) in three of six years for the spring preplant treatment. Averaged across the six rotation cycles, flow-normalized NO3-N losses ranked in the order: fall N > spring N + NP > fall N + NP > spring N. Under these conditions, NO3 losses in subsurface drainage from a corn-soybean rotation can be reduced 14% by spring N and 10% by late fall N + NP compared with fall-applied N. Nitrate losses were not appreciably reduced by adding NP to spring preplant N. PMID- 15758113 TI - Phosphorus in fresh and dry dung of grazing dairy cattle, deer, and sheep: sequential fraction and phosphorus-31 nuclear magnetic resonance analyses. AB - Knowledge of phosphorus (P) fractions in dung of animals (dairy cattle, deer, sheep) grazing pasture is important for soil fertility and the potential for P transport in runoff and subsequent surface water quality deterioration. We used sequential fractionation and 31P nuclear magnetic resonance (NMR) spectroscopy to determine P forms in fresh and air-dried (to simulate field conditions during grazing) dung. Sheep dung was richest in P (8 g kg(-1)), and cattle dung poorest (5.5 g kg(-1)). Data for sequential fractionation indicated that most P was extractable by water (15-36%) and bicarbonate (36-45%) in fresh dung, and shifted toward recalcitrant, HCl (12-28%), and residual P forms (15-31%) with drying. Organic P concentration in dung was poor (maximum of 15% of total P), probably due to the poor concentration of phytate in pasture. The 31P NMR spectra of NaOH EDTA extracts supported this by detecting a low concentration of monoesters (9 19% of total P in extracts), of which phytate is a major component. The 31P NMR data also showed that changes in organic P concentration with drying could be due to the degradation of diesters. Data indicate the decreasing bioavailability of dairy cattle, deer, and sheep dung with drying and the need to consider this effect with respect to P returns for soil fertility and the potential for runoff. PMID- 15758114 TI - Leaching of glyphosate and amino-methylphosphonic acid from Danish agricultural field sites. AB - Pesticide leaching is an important process with respect to contamination risk to the aquatic environment. The risk of leaching was thus evaluated for glyphosate (N-phosphonomethyl-glycine) and its degradation product AMPA (amino methylphosphonic acid) under field conditions at one sandy and two loamy sites. Over a 2-yr period, tile-drainage water, ground water, and soil water were sampled and analyzed for pesticides. At a sandy site, the strong soil sorption capacity and lack of macropores seemed to prevent leaching of both glyphosate and AMPA. At one loamy site, which received low precipitation with little intensity, the residence time within the root zone seemed sufficient to prevent leaching of glyphosate, probably due to degradation and sorption. Minor leaching of AMPA was observed at this site, although the concentration was generally low, being on the order of 0.05 microg L(-1) or less. At another loamy site, however, glyphosate and AMPA leached from the root zone into the tile drains (1 m below ground surface [BGS]) in average concentrations exceeding 0.1 microg L(-1), which is the EU threshold value for drinking water. The leaching of glyphosate was mainly governed by pronounced macropore flow occurring within the first months after application. AMPA was frequently detected more than 1.5 yr after application, thus indicating a minor release and limited degradation capacity within the soil. Leaching has so far been confined to the depth of the tile drains, and the pesticides have rarely been detected in monitoring screens located at lower depths. This study suggests that as both glyphosate and AMPA can leach through structured soils, they thereby pose a potential risk to the aquatic environment. PMID- 15758115 TI - Simulating solute transport in a structured field soil: uncertainty in parameter identification and predictions. AB - Dual-permeability models have been developed to account for the significant effects of macropore flow on contaminant transport, but their use is hampered by difficulties in estimating the additional parameters required. Therefore, our objective was to evaluate data requirements for parameter identification for predictive modeling with the dual-permeability model MACRO. Two different approaches were compared: sequential uncertainty fitting (SUFI) and generalized likelihood uncertainty estimation (GLUE). We investigated six parameters controlling macropore flow and pesticide sorption and degradation, applying MACRO to a comprehensive field data set of bromide andbentazone [3-isopropyl-1H-2,1,3 benzothiadiazin-4(3H)-one-2,2dioxide] transport in a structured soil. The GLUE analyses of parameter conditioning for different combinations of observations showed that both resident and flux concentrations were needed to obtain highly conditioned and unbiased parameters and that observations of tracer transport generally improved the conditioning of macropore flow parameters. The GLUE "behavioral" parameter sets covered wider parameter ranges than the SUFI posterior uncertainty domains. Nevertheless, estimation uncertainty ranges defined by the 5th and 95th percentiles were similar and many simulations randomly sampled from the SUFI posterior uncertainty domains had negative model efficiencies (minimum of -3.2). This is because parameter correlations are neglected in SUFI and the posterior uncertainty domains were not always determined correctly. For the same reasons, uncertainty ranges for predictions of bentazone losses through drainflow for good agricultural practice in southern Sweden were 27% larger for SUFI compared with GLUE. Although SUFI proved to be an efficient parameter estimation tool, GLUE seems better suited as a method of uncertainty estimation for predictions. PMID- 15758116 TI - Land application of domestic effluent onto four soil types: plant uptake and nutrient leaching. AB - Land application has become a widely applied method for treating wastewater. However, it is not always clear which soil-plant systems should be used, or why. The objectives of our study were to determine if four contrasting soils, from which the pasture is regularly cut and removed, varied in their ability to assimilate nutrients from secondary-treated domestic effluent under high hydraulic loadings, in comparison with unirrigated, fertilized pasture. Grassed intact soil cores (500 mm in diameter by 700 mm in depth) were irrigated (50 mm wk(-1)) with secondary-treated domestic effluent for two years. Soils included a well-drained Allophanic Soil (Typic Hapludand), a poorly drained Gley Soil (Typic Endoaquept), a well-drained Pumice Soil formed from rhyolitic tephra (Typic Udivitrand), and a well-drained Recent Soil formed in a sand dune (Typic Udipsamment). Effluent-irrigated soils received between 746 and 815 kg N ha(-1) and 283 and 331 kg P ha(-1) over two years of irrigation, and unirrigated treatments received 200 kg N ha(-1) and 100 kg P ha(-1) of dissolved inorganic fertilizer over the same period. Applying effluent significantly increased plant uptake of N and P from all soil types. For the effluent-irrigated soils plant N uptake ranged from 186 to 437 kg N ha(-1) yr(-1), while plant P uptake ranged from 40 to 88 kg P ha(-1) yr(-1) for the effluent-irrigated soils. Applying effluent significantly increased N leaching losses from Gley and Recent Soils, and after two years ranged from 17 to 184 kg N ha(-1) depending on soil type. Effluent irrigation only increased P leaching from the Gley Soil. All P leaching losses were less than 49 kg P ha(-1) after two years. The N and P leached from effluent treatments were mainly in organic form (69-87% organic N and 35-65% unreactive P). Greater N and P leaching losses from the irrigated Gley Soil were attributed to preferential flow that reduced contact between the effluent and the soil matrix. Increased N leaching from the Recent Soil was the result of increased leaching of native soil organic N due to the higher hydraulic loading from the effluent irrigation. PMID- 15758117 TI - Cattle feedlot soil moisture and manure content: I. Impacts on greenhouse gases, odor compounds, nitrogen losses, and dust. AB - Beef cattle feedlots face serious environmental challenges associated with manure management, including greenhouse gas, odor, NH3, and dust emissions. Conditions affecting emissions are poorly characterized, but likely relate to the variability of feedlot surface moisture and manure contents, which affect microbial processes. Odor compounds, greenhouse gases, nitrogen losses, and dust potential were monitored at six moisture contents (0.11, 0.25, 0.43, 0.67, 1.00, and 1.50 g H2O g(-1) dry matter [DM]) in three artificial feedlot soil mixtures containing 50, 250, and 750 g manure kg(-1) total (manure + soil) DM over a two week period. Moisture addition produced three microbial metabolisms: inactive, aerobic, and fermentative at low, moderate, and high moisture, respectively. Manure content acted to modulate the effect of moisture and enhanced some microbial processes. Greenhouse gas (CO2, N2O, and CH4) emissions were dynamic at moderate to high moisture. Malodorous volatile fatty acid (VFA) compounds did not accumulate in any treatments, but their persistence and volatility varied depending on pH and aerobic metabolism. Starch was the dominant substrate fueling both aerobic and fermentative metabolism. Nitrogen losses were observed in all metabolically active treatments; however, there was evidence for limited microbial nitrogen uptake. Finally, potential dust production was observed below defined moisture thresholds, which were related to manure content of the soil. Managing feedlot surface moisture within a narrow moisture range (0.2-0.4 g H2O g(-1) DM) and minimizing the accumulation of manure produced the optimum conditions that minimized the environmental impact from cattle feedlot production. PMID- 15758118 TI - Cattle feedlot soil moisture and manure content: II. Impact on Escherichia coli O157. AB - The moisture and manure contents of soils at cattle feedlot surfaces vary spatiotemporally and likely are important factors in the persistence of Escherichia coli O157 in these soils. The impacts of water content (0.11-1.50 g H2O g(-1) dry feedlot surface material [FSM]) and manure level (5, 25, and 75% dry manure in dry FSM) on E. coli O157:H7 in feedlot soils were evaluated. Generally, E. coli O157:H7 numbers either persisted or increased at all but the lowest moisture levels examined. Manure content modulated the effect of water on E. coli growth; for example, at water content of 0.43 g H2O g(-1) dry FSM and 25% manure, E. coli O157:H7 increased by 2 log10 colony forming units (CFU) g(-1) dry FSM in 3 d, while at 0.43 g H2O g(-1) dry FSM and 75% manure, populations remained stable over 14 d. Escherichia coli and coliform populations responded similarly. In a second study, the impacts of cycling moisture levels and different drying rates on naturally occurring E. coli O157 in feedlot soils were examined. Low initial levels of E. coli O157 were reduced to below enumerable levels by 21 d, but indigenous E. coli populations persisted at >2.50 log10 CFU g(-1) dry FSM up to 133 d. We conclude that E. coli O157 can persist and may even grow in feedlot soils, over a wide range of water and manure contents. Further investigations are needed to determine if these variables can be manipulated to reduce this pathogen in cattle and the feedlot environment. PMID- 15758119 TI - Modeling carbon and nitrogen transformations for adjustment of compost application with nitrogen uptake by wheat. AB - Environmentally sound management of the use of composts in agriculture relies on matching the rate of release of available N from compost-amended soils to the crop demand. To develop such management it is necessary to (i) characterize the properties of composts that control their rates of decomposition and release of N and (ii) determine the optimal amount of composts that should be applied annually to wheat (Triticum aestivum L.). Carbon and N mineralization were measured under controlled conditions to determine compost decomposition rate parameters, and the NCSOIL model was used to derive the organic wastes parameters that control the rates of N and C transformations in the soil. We also characterized the effect of a drying period to estimate the effects of the dry season on C and N dynamics in the soil. The optimized compost parameters were then used to predict mineral N concentration dynamics in a soil-wheat system after successive annual applications of compost. Sewage sludge compost (SSC) and cattle manure compost (CMC) mineralization characteristics showed similar partitioning into two components of differing ease of decomposition. The labile component accounted for 16 to 20% of total C and 11 to 14% of total N, and it decomposed at a rate of 2.4 x 10(-2) d(-1), whereas the resistant pool had a decomposition rate constant of 1.2 to 1.4 x 10(-4) d(-1). The main differences between the two composts resulted from their total C and N and inorganic N contents, which were determined analytically. The long-term effect of a drying period on C and N mineralization was negligible. Use of these optimization results in a simulation of compost mineralization under a wheat crop, with a modified plant-effect version of the NCSOIL model, enabled us to evaluate the effects of the following factors on the C and N dynamics in soil: (i) soil temperature, (ii) mineral N uptake by plants, and (iii) release of very labile organic C in root exudates. This labile organic C enhanced N immobilization following application, and so decreased the N available for uptake by plants. PMID- 15758120 TI - Chemical and physical properties of dry flue gas desulfurization products. AB - Beneficial and environmentally safe recycling of flue gas desulfurization (FGD) products requires detailed knowledge of their chemical and physical properties. We analyzed 59 dry FGD samples collected from 13 locations representing four major FGD scrubbing technologies. The chemistry of all samples was dominated by Ca, S, Al, Fe, and Si and strong preferential partitioning into the acid insoluble residue (i.e., coal ash residue) was observed for Al, Ba, Be, Cr, Fe, Li, K, Pb, Si, and V. Sulfur, Ca, and Mg occurred primarily in water- or acid soluble forms associated with the sorbents or scrubber reaction products. Deionized water leachates (American Society for Testing and Materials [ASTM] method) and dilute acetic acid leachates (toxicity characteristic leaching procedure [TCLP] method) had mean pH values of >11.2 and high mean concentrations of S primarily as SO(2-)4 and Ca. Concentrations of Ag, As, Ba, Cd, Cr, Hg, Pb, and Se (except for ASTM Se in two samples) were below drinking water standards in both ASTM and TCLP leachates. Total toxicity equivalents (TEQ) of dioxins, for two FGD products used for mine reclamation, were 0.48 and 0.53 ng kg(-1). This was similar to the background level of the mine spoil (0.57 ng kg(-1)). The FGD materials were mostly uniform in particle size. Specific surface area (m2 g(-1)) was related to particle size and varied from 1.3 for bed ash to 9.5 for spray dryer material. Many of the chemical and physical properties of these FGD samples were associated with the quality of the coal rather than the combustion and SO2 scrubbing processes used. PMID- 15758121 TI - Phosphorus speciation in broiler litter and turkey manure produced from modified diets. AB - Modifying poultry diets by reducing mineral P supplementation and/or adding phytase may change the chemical composition of P in manures and affect the mobility of P in manure-amended soils. We studied the speciation of P in manures produced by broiler chickens and turkeys from either normal diets, or diets with reduced amounts of non-phytate phosphorus (NPP) and/or phytase, using a combination of chemical fractionation and synchrotron X-ray absorption near edge structure (XANES) spectroscopy. All broiler litters were rich in dicalcium phosphate (65-76%), followed by aqueous phosphate (13-18%), and phytic acid (7 20%); however, no hydroxylapatite was observed. Similarly, normal turkey manure had 77% of P as dicalcium phosphate and had no hydroxylapatite, while turkey manure from diets that had reduced NPP and phytase contained equal proportions of dicalcium phosphate (33-45%) and hydroxylapatite (35-39%). This is attributed to the higher total Ca to P ratio (>2) in modified turkey manures that resulted in transformation of more soluble (dicalcium phosphate) to less soluble P compounds (hydroxylapatite). Chemical fractionation showed that H2O-extractable P was the predominant form in broiler litter (56-77%), whereas aqueous phosphate determined with XANES was <18% indicating that H2O probably dissolved mineral forms of P (e.g., dicalcium phosphate). Results show that HCl extraction primarily removed phytic acid from broiler litters and normal turkey manure, while it removed a mixture of hydroxylapatite and phytic acid from modified turkey manures. The combination of chemical fractionation and XANES provided information about the nature of P in these manures, which may help to devise best management practices for manure use. PMID- 15758122 TI - Gaseous nitrogen and carbon losses from pig manure derived from different diets. AB - Manipulation of the diets of pigs may alter the composition of the manure and thereby the environmental and agricultural qualities of the manure. Laboratory studies were performed to quantify the effect of manipulation of pig diets on the chemical composition of the derived manure (slurry), the potential emission of methane (CH4) and ammonia (NH3) during anaerobic storage of the manure, and the potential nitrous oxide (N2O) and carbon dioxide (CO2) emission after application of the manure to soil. The diets differed in contents of crude protein and salt (CaSO4), and the type and contents of nonstarch polysaccharides (NSP). Emissions of NH3 and CH4 during storage were smaller at a low than at a high dietary protein content. The emission of NH3 was significantly related to the contents of ammonium (NH4), total N, and pH. The emission of CH4 was significantly related to contents of dry matter, total C, and volatile fatty acids in the manure. The effect of manure composition on N2O emission markedly differed between the two tested soils, which points at interactions with soil properties such as the organic matter content. These types of interactions require soil-specific recommendations for mitigation of N2O emission from soil-applied pig manure by manipulation of the diet. From the tested diets, decreasing the protein content has the largest potential to simultaneously decrease NH3 and CH4 emissions during manure storage and N2O emission from soil. An integral assessment of the environmental and agricultural impact of handling and application of pig manure as a result of diet manipulation provides opportunities for farmers to maximize the value of manures as fertilizer and soil conditioner and to minimize N and C emissions to the environment. PMID- 15758123 TI - A methodology to estimate the denitrifying capacity of a riparian wetland. AB - Numerous studies have shown that riparian wetlands can play an important role in reducing nitrate concentrations before the ground water discharges into streams. Denitrification has been identified as an important process for this removal. Several approaches have been proposed to predict the denitrifying removal capacity of a riparian wetland, but no widely used tool exists to precisely quantify this capacity at the landscape scale. We propose such a methodology based on modeling the spatial variation of soil-water interactions in the entire riparian wetland. Mean values of denitrification enzyme activity (DEA) within three soil-denitrifying classes were 604, 212, and 24 ng N g(-1) h(-1) for Classes 3, 2, and 1, respectively. The study area, having a ground surface of about 15000 m2, was underlain by an aquifer with a calculated volume of 60000 m3, less than 10000 m3 of which corresponded to active denitrifying horizons (Classes 2 and 3). By volume, approximately 30% of Class 3 and 70% of Class 2 were interacting with ground water. The denitrifying removal capacity of our wetland was calculated to be about 1.8 kg N m(-2) yr(-1). The calculated denitrifying capacity of our site was less than expected. This is due to the fact that not all ground water interacts with the horizons having the highest denitrifying capacity. Thus, we show that whatever the system is, specific local pedological and hydrogeological conditions and their interactions are paramount in controlling the denitrification process. PMID- 15758124 TI - Cascading ecological effects of low-level phosphorus enrichment in the Florida everglades. AB - Few studies have examined long-term ecological effects of sustained low-level nutrient enhancement on wetland biota. To determine sustained effects of phosphorus (P) addition on Everglades marshes we added P at low levels (5, 15, and 30 microg L(-1) above ambient) for 5 yr to triplicate 100-m flow-through channels in pristine marsh. A cascade of ecological responses occurred in similar sequence among treatments. Although the rate of change increased with dosing level, treatments converged to similar enriched endpoints, characterized most notably by a doubling of plant biomass and elimination of native, calcareous periphyton mats. The full sequence of biological changes occurred without an increase in water total P concentration, which remained near ambient levels until Year 5. This study indicates that Everglades marshes have a near-zero assimilative capacity for P without a state change, that ecosystem responses to enrichment accumulate over time, and that downstream P transport mainly occurs through biota rather than the water column. PMID- 15758125 TI - Phosphorus flux from bottom sediments in Lake Eucha, Oklahoma. AB - Phosphorus inputs into reservoirs include external sources from the watershed and internal sources from the reservoir bottom sediments. This study quantified sediment P flux in Lake Eucha, northeastern Oklahoma, USA, and evaluated the effectiveness of chemical treatment to reduce sediment P flux. Six intact sediment-water columns were collected from three sites in Lake Eucha near the reservoir channel at depths of 10 to 15 m. Three intact sediment and water columns from each site were incubated for 21 d at approximately 22 degrees C under aerobic conditions, and three were incubated under anaerobic conditions (N2 with 300 ppm CO2); sediment P flux was estimated over the 21 d for each core. The overlying water in the cores was bubbled with air for approximately 1 wk and then treated with aluminum sulfate (alum). The cores were incubated at approximately 22 degrees C for an additional 14 d under aerobic or anaerobic conditions, and sediment P flux after alum treatment was estimated for each core. Sediment P flux was approximately four times greater under anaerobic conditions compared with aerobic conditions. Alum treatment of the intact sediment-water columns reduced (8x) sediment P flux under anaerobic conditions. Internal P flux (1.03 and 4.40 mg m(-2) d(-1) under aerobic and anaerobic conditions, respectively) was greater than external P flux (0.13 mg m(-2) d(-1)). The internal P load (12 Mg yr(-1)) from reservoir bottom sediments was almost 25% of the external P load (approximately 48 Mg yr(-1)) estimated using a calibrated watershed model. PMID- 15758126 TI - Sirolimus-eluting stents versus the superficial femoral artery: second round. PMID- 15758127 TI - Thrombolytic therapy for acute superior mesenteric artery occlusion. AB - The aim of this review is to evaluate thrombolytic therapy for acute superior mesenteric artery occlusion as an alternative or adjunctive treatment modality to surgical therapy and to provide current knowledge for timely and informed decisions regarding treatment of acute mesenteric ischemia. A systematic analysis of the available literature from 1966 to 2003 regarding thrombolytic therapy for superior mesenteric artery thromboembolism was performed. A total of 20 case reports and seven small series covered 48 patients with acute superior mesenteric artery thromboembolism. In the herein reviewed series, thrombolytic therapy of acute superior mesenteric artery thromboembolism resulted in angiographic resolution of the thromboembolism in 43 patients, in clinical success without requiring additional surgical intervention in 30 patients, and in survival in 43 patients, with similar complication rates as in thrombolytic treatment of peripheral vascular occlusions. Remission of abdominal pain during the first few hours of treatment formed the most important indicator of therapeutic success. Insufficient evidence from reviewed literature is available to determine the relative effectiveness and safety of thrombolytic treatment for acute superior mesenteric artery thromboembolism; however, initial results appear to be promising. Thrombolytic therapy can be effective relatively quickly, may obviate surgery, and has the potential to resolve the clot completely. In some cases it can be used as an alternative or neo-adjunctive treatment modality to surgery. A treatment guideline for thrombolysis of acute superior mesenteric artery thromboembolism should be developed. PMID- 15758129 TI - Case-controlled comparison of a percutaneous collagen arteriotomy closure device versus manual compression after liver chemoembolization. AB - PURPOSE: To compare complications and outcomes between the use of the Duett collagen closure device after one or multiple deployments and manual compression in patients treated with transcatheter arterial chemoembolization (TACE) for primary or metastatic liver cancer. MATERIALS AND METHODS: A database of 214 patients who underwent a total of 426 TACE procedures between July 2001 and July 2003 was retrospectively analyzed with regard to the use of the Duett closure device to obtain hemostasis. The Duett device was used in 211 cases (121 patients), whereas manual compression was performed in 215 cases (93 patients). Primary endpoints included complications related to hemostasis, time to hemostasis (TTH), time to ambulation (TTA), and time to discharge (TTD). Risk factors were tested for correlation with complications (P < .05). Other endpoints included descriptive data regarding the Duett treatment group. RESULTS: Only minor complications were observed in both groups, without a statistically significant difference (P = .16). The mean TTH and TTA were significantly shorter (P < .0001) in the Duett group, whereas there was no difference in TTD between groups (P = .59). Reaccessing the same arterial site for separate procedures was not a significant risk factor for complications in the Duett group (P < .0001). CONCLUSION: The Duett closure device achieves similar safety and efficacy as manual compression in this distinct group of patients. In addition, this device can be safely and repetitively deployed at the arteriotomy site after each TACE procedure. PMID- 15758130 TI - The safety and efficacy of a percutaneous closure device in patients undergoing uterine artery embolization. AB - PURPOSE: The use of suture-mediated closure devices (SMCDs) in patients undergoing uterine artery embolization (UAE) for symptomatic leiomyomata is controversial. With recent literature suggesting a higher complication rate with the use of SMCDs, their use in this specific population has been questioned. The purpose of this study is to assess the safety and efficacy of SMCDs specifically for those patients undergoing UAE for symptomatic uterine leiomyomata. MATERIALS AND METHODS: A prospective database was established in a single high-volume medical center for patients undergoing UAE for symptomatic leiomyomata. Data were tracked prospectively, with technical outcome and complication rates serving as endpoints. RESULTS: UAE was performed in 342 consecutive patients from January 2001 to September 2003. The SMCD was used in 328 of these patients (96%). Successful primary hemostasis was achieved in 320 of 328 patients (97%), with additional manual compression required in the remaining eight patients (3%). No major complications were observed. Minor complications consisted of minor hematomas in four women (1%) and anteriomedial thigh pain in 68 women (21%) within 1 month of follow-up, resulting in an overall complication rate of 22%. All symptoms related to anteromedial thigh pain ipsilateral to the arterial puncture site were managed conservatively with 1 week of oral nonsteroidal antiinflammatory medication. CONCLUSION: The SMCD provided safe and effective hemostasis in patients undergoing UAE. Transient thigh discomfort, which may result from irritation of the anterior femoral cutaneous nerves (femoral neuralgia syndrome), was uniformly relieved with nonsteroidal antiinflammatory therapy. PMID- 15758128 TI - Sirolimus-eluting versus bare nitinol stent for obstructive superficial femoral artery disease: the SIROCCO II trial. AB - PURPOSE: To investigate further the safety and efficacy of the sirolimus-eluting S. M.A.R.T. Nitinol Self-expanding Stent by comparison with a bare stent in superficial femoral artery (SFA) obstructions. MATERIALS AND METHODS: This randomized, double-blind study involved 57 patients (29 in the sirolimus-eluting stent group and 28 in the bare stent group) with chronic limb ischemia and SFA occlusions (66.7%) or stenoses (average lesion length, 81.5 mm +/- 41.2). Stent implantation followed standard interventional techniques and a maximum of two stents could be implanted. The primary endpoint was the in-stent mean lumen diameter at 6 months as determined by quantitative angiography. RESULTS: Both stent types were effective in revascularizing the diseased SFA and allowing sustained patency for at least 6 months. There was no statistically significant difference between treatment groups in the in-stent mean lumen diameter at 6 months (4.94 mm +/- 0.69 and 4.76 mm +/- 0.54 mm for sirolimus-eluting and bare stent groups, respectively; P = .31). Although the diameter of the target lesion tended to be larger and percent stenosis tended to be lower with the sirolimus eluting stent, there were no statistically significant differences between treatments in terms of any of the variables. The mean late loss values were 0.38 mm +/- 0.64 and 0.68 mm +/- 0.97 for the sirolimus-eluting stent group and the bare stent group, respectively (P = .20). The binary restenosis rates, with a cutoff of 50% at 6 months, were zero in the sirolimus-eluting stent group and 7.7% in the bare stent group (P = .49). Clinical outcomes matched angiographic outcomes with improvements in ankle-brachial index and symptoms of claudication. There was no significant difference between treatments in terms of adverse events. CONCLUSION: Although there is a trend for greater efficacy in the sirolimus-eluting stent group, there were no statistically significant differences in any of the variables. PMID- 15758131 TI - Transcatheter arterial chemoembolization in unresectable cholangiocarcinoma: initial experience in a single institution. AB - PURPOSE: Unresectable cholangiocarcinoma carries a dismal prognosis, with median survival times ranging from 6 to 12 months from the time of diagnosis. Palliative therapies have been disappointing and have not been shown to significantly prolong survival. Conversely, transcatheter arterial chemoembolization (TACE) has been effective in prolonging the lives of patients with hepatocellular carcinoma but has not been used against cholangiocarcinoma. Therefore, the purpose of the present study was to assess the safety and efficacy (ie, survival) of TACE in patients with unresectable intrahepatic cholangiocarcinoma. MATERIALS AND METHODS: Seventeen patients with unresectable cholangiocarcinoma were treated with one or more cycles of TACE between 1995 and 2004 at our institution. Follow up imaging was performed on all patients 4-6 weeks after each TACE procedure to determine tumor response and need for further treatment. Survival was calculated with use of the Kaplan-Meier survival curve. RESULTS: The median survival for 17 patients treated with TACE was 23 months. Two patients with previously unresectable disease underwent successful resection after TACE. The procedure was well tolerated by 82% of the patients, who experienced no side effects or mild side effects that quickly resolved with conservative therapy alone. Two patients had minor complications (12%), which were managed successfully, and one had a major complication that resulted in a fatal outcome. This patient had a rapidly declining course from the time of diagnosis and died shortly after TACE. CONCLUSIONS: The results suggest that TACE was effective at prolonging survival of patients with unresectable cholangiocarcinoma. Therefore, for these patients, TACE may be an appropriate palliative therapy. PMID- 15758132 TI - Peritoneal ports for treatment of intractable ascites. AB - PURPOSE: To present clinical data for a new peritoneal port for minimally invasive treatment of intractable ascites that can be used for aspiration in a patient's home. MATERIALS AND METHODS: Twenty-eight consecutive peritoneal ports were placed in 27 patients with intractable ascites. Ascites etiology was malignancy in 22 patients, cirrhosis in three, pancreatic duct injury in one, and unknown in one. Technical and clinical success and complications were evaluated until the time of death or the end of the study. RESULTS: All ports were inserted successfully with removal of all ascites, and all patients had immediate and complete symptom relief. Ascites was managed by periodic drainage, typically by a visiting nurse in the patient's home. The long-term clinical success rate was 96%, with 26 of 27 patients exhibiting maintained relief of symptoms until death or the end of the study. The long-term patency rate was 100% after 1,810 patient days. Only one patient (4%) had a major complication. This was a port leak that required port exchange. Subsequently, the patient developed bacterial peritonitis. CONCLUSION: Peritoneal ports appear to be a safe, effective, minimally invasive treatment for intractable ascites. This device allows for reliable ascites aspiration in the patient's home. PMID- 15758133 TI - Cross-species pharmacologic evaluation of plasmin as a direct-acting thrombolytic agent: ex vivo evaluation for large animal model development. AB - PURPOSE: Human plasma-derived plasmin has been developed for the treatment of thrombosed hemodialysis arteriovenous grafts and vascular occlusive diseases. To further investigate this drug in large animal models and derive preliminary dosing estimates, the authors compared plasmin's relative lytic potential in four species, including man. The goal was to find which species' whole blood clots best compared to human clots in terms of lysis with plasmin. The results from these studies will serve to guide species selection for large animal experimentation. MATERIALS AND METHODS: Clotted blood from human, pig, sheep, and bovine subjects were treated with saline solution control, plasmin, or tissue plasminogen activator. Electron microscopy (EM) techniques were used to investigate the effects of clot size and fragmentation on plasmin lysis, the effects of intrathrombic infusion by injection of plasmin directly into whole blood clots, and species fibrin structural differences. RESULTS: Under static conditions, plasmin efficiently lysed clots from all species studied at an optimal dose of 4-5 mg per 4-5 g of clot. With fragmented human clots, plasmin (5 mg)-induced lysis was 80% +/- 2% at 60 minutes. Porcine clots were more resistant to plasmin lysis compared with human, ovine, and bovine clots. Percent lysis at 60 minutes with plasmin for ovine clots was 72% +/- 3% (4-mg dose), compared with 50% +/- 4% for porcine clots (5-mg dose; P < .05). EM of porcine clots showed a compact fibrin network that appeared more dense than that in human or sheep clots, which may account for the decreased lytic rate. CONCLUSIONS: Human plasmin is an effective direct-acting thrombolytic agent that is capable of lysing fibrin from several species. Ex vivo lysis studies were used to investigate the most appropriate large animal model that best approximates plasmin lysis with human clots under certain conditions. It was determined that ovine clots treated with plasmin most closely resemble the lysis observed with human clots. PMID- 15758135 TI - Alteplase stability and bioactivity after low-power ultrasonic energy delivery with the OmniSonics resolution system. AB - PURPOSE: Low-power ultrasonic (US) energy is capable of clot dissolution in vivo. The combination of US energy plus alteplase may further accelerate clot lysis; however, the effects of cavitation could potentially denature and inactivate the lytic protein. The purpose of this study was to determine the bioactivity and stability of alteplase when exposed to US energy with use of a novel intravascular US wire in an in vitro model. MATERIALS AND METHODS: The model consisted of a 6.4-mm-diameter silicone tube closed at one end and filled with alteplase (1 mg/mL) in a water bath (37 degrees C). A 95-cm US wire (0.025-inch diameter, 20 kHz) was inserted into the tube and connected to a variable power generator. The wire delivers low-power acoustic energy 360 degrees around its 20 cm active length and was irrigated by a continuous infusion of purified water. Fresh 6-mL alteplase aliquots were exposed to US energy and tested in duplicates. Zero (control), 1 W, or 2 W of energy was delivered to individual test samples for zero (control), 0.5, 3, or 6 minutes. Alteplase samples were assayed for optical clarity and protein concentration with use of UV spectrophotometry, for percent protein monomer with use of high-performance size-exclusion chromatography, and for in vitro clot lysis activity. RESULTS: In the control samples, optical clarity was clear or colorless in all samples; protein concentration was 1.02 mg/mL +/- 0; protein monomer was 98%; and clot lysis activity was 108% per mg +/- 1. In the test samples, optical clarity was clear or colorless in all samples; protein concentrations at 0.5, 3, and 6 minutes were 0.98 mg/mL +/- 0.02, 0.93 mg/mL +/- 0.01, and 0.86 mg/mL +/- 0.02, respectively, at 1 W, and 1.00 mg/mL +/- 0.03, 0.94 mg/mL +/- 0.10, and 0.84 mg/mL +/- 0.17, respectively, at 2 W. Protein monomer was 98% for all samples. Clot lysis activity levels at 0.5, 3, and 6 minutes were 111% per mg +/- 1, 110% per mg +/- 1, and 115% per mg +/- 1, respectively, at 1 W, and 110% per mg +/- 0, 111% per mg +/- 1, and 116% per mg +/- 2, respectively, at 2 W. CONCLUSIONS: Alteplase solutions exposed to low-power US energy for as long as 6 minutes remained fully active and stable as determined by protein assays. Further investigation is warranted with use of combinations of US energy and alteplase. PMID- 15758134 TI - Alteplase as a catheter locking solution: in vitro evaluation of biochemical stability and antimicrobial properties. AB - PURPOSE: To reduce potential complications of fibrin deposition to catheter surfaces, there is increasing empiric use of alteplase as a catheter lock solution. The purpose is to evaluate the properties of alteplase when reconstituted in sterile water (SW) or bacteriostatic water (BW) for prolonged periods. MATERIALS AND METHODS: Alteplase in glass vials was reconstituted (1 mg/mL) with SW or BW (0.9% benzyl alcohol) in duplicates and stored at 37 degrees C. Biochemical assays were performed at days 0 and 7 and included optical clarity, protein concentration, percent protein monomer, and in vitro clot lysis activity. Microbiologic assays were performed on days 7 through 28 with use of a standardized antimicrobial effectiveness test (pass/fail) and pour-plate methods incubated at 22.5 degrees C (fungus, 3-7 days) or 32.5 degrees C (bacteria, 3-5 days). Organisms tested included Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Aspergillus niger. RESULTS: Biochemical assay results were as follows: on day 0, all samples were clear/colorless; protein concentrations were 1.10 mg/mL +/- 0 in SW and 1.11 mg/mL +/- 0 in BW; percent protein monomer was 8.2% +/- 0.07 in SW and 98.6% +/- 0.07 in BW; and in vitro clot lysis activity (in percent of relative activity) was 100% in all samples. On day 7, all samples were clear/colorless, protein concentrations were 1.11 mg/mL +/- 0.07 in SW and 1.11 mg/mL +/- 0.07 in BW; percent protein monomer was 97.4% +/- 0.21 in SW and 96.1% +/- 0.21 in BW; and in vitro clot lysis activity (relative activity compared with day 0) was 91% +/- 2.8 in SW and 90% +/- 2.8 in BW. Microbiologic assays (US Pharmacopeia [USP] antimicrobial effectiveness test) yielded a failing result for alteplase reconstituted in SW and a passing result for alteplase reconstituted in BW. CONCLUSIONS: Alteplase reconstituted with SW or BW remains relatively stable with retained bioactivity when stored at 37 degrees C for as long as 7 days. Despite the biochemical similarities of the two solutions, only alteplase in BW met USP criteria as an effective antimicrobial solution. Further clinical evaluation is warranted. PMID- 15758136 TI - Removal of Gunther Tulip vena cava filter through femoral vein approach. AB - The Gunther Tulip vena cava filter is designed for removal by the internal jugular vein approach with use of a blunted hook placed at the superior aspect of the filter. Removal of this filter was performed by the femoral approach in a patient with central venous occlusion that precluded removal by the conventional approach. PMID- 15758137 TI - Inferior vena cava filter removal after 317-day implantation. AB - A Gunther Tulip inferior vena cava (IVC) filter was placed prophylactically in a 22-year-old trauma victim with spinal injuries. Attempts to retrieve the filter at 21 and 25 days after placement were aborted as a result of clot trapped in the filter. Despite the possible risk of an IVC laceration, a third attempt was made 317 days after placement in view of the young age of the patient. The filter started collapsing into the retrieval sheath but could not easily be separated from the IVC. During attempted redeployment, the filter would not reopen. The filter was ultimately retrieved with use of additional force. A mild stenosis of the IVC was noted immediately after retrieval. However, the IVC returned to its preretrieval diameter as seen on a 3-month follow-up venogram. PMID- 15758138 TI - Percutaneous salvage of a failing rex shunt. AB - Creation of a mesenterico-left portal vein (PV) shunt with use of autologous internal jugular vein (ie, Rex shunt) is a surgical option for the treatment of symptomatic extrahepatic PV occlusion. Herein a patient is described who underwent angioplasty and stent placement across a shunt stenosis by ultrasound (US)--guided percutaneous transhepatic portal access. Follow-up US has demonstrated continued shunt patency. PMID- 15758139 TI - Radiofrequency ablation of a recanalized renal arteriovenous malformation. AB - Radiofrequency (RF) ablation is becoming established as a method of treatment of small renal neoplasms. Successful RF ablation of a renal arteriovenous malformation (AVM) was performed in a patient who had previously undergone coil embolization and experienced hematuria after the feeding vessels recanalized. The 3.5-cm lesion was successfully treated with RF ablation, resulting in cessation of hematuria. At follow-up, there was no residual enhancement of the AVM on computed tomography and the lesion had decreased in size. PMID- 15758140 TI - Bronchobiliary fistula after radiofrequency thermal ablation of hepatic tumor. AB - A broad spectrum of complications can occur after radiofrequency (RF) ablation of hepatic tumors, even though it has been accepted as a safe and effective technique for unresectable hepatic tumors. Recently, the rare complication of brochobiliary fistula was encountered after RF ablation in a patient with a metastatic tumor from stomach cancer. It was assumed to have developed from collateral damage to the adjacent diaphragm and lung base as well as biloma formation at the ablation zone. Symptomatic improvement was achieved by conservative management with an external drainage catheter, but the fistula was still persistent on a 2-month follow-up image. PMID- 15758141 TI - CT and fluoroscopically guided percutaneous embolization treatment of a pseudoaneurysm associated with pancreatitis. AB - A novel approach to embolization treatment with a cyanoacrylate solution for a bleeding pseudoaneurysm associated with pancreatitis is described in the setting of a critically ill patient. Computed tomography and a C-arm fluoroscopic unit were used to guide percutaneous placement of a needle and injection of the embolic material. This technique represents an alternative form of embolization when traditional angiographic techniques are not possible. PMID- 15758142 TI - Radiofrequency ablation for tumor-related massive hematuria. AB - To determine whether radiofrequency (RF) ablation targeting the tumor-collecting system interface has a durable effect in patients with transfusion-dependent kidney tumor-related hematuria, four patients aged 61-71 years were successfully treated with RF ablation, with a mean follow up of 12 months. Baseline creatinine levels varied from 2.0 mg/dL to 3.7 mg/dL. All patients had received red blood cell transfusions in the days and hours before RF ablation. No subsequent surgical or interventional procedures were required for management of hematuria. Gross hematuria resolved in 24-48 hours in all four patients. Two of the patients are alive with stable renal function and two died of causes unrelated to treatment. RF ablation may be an effective therapeutic option for transfusion dependent cancer-related hematuria in patients with renal insufficiency, solitary kidney, or comorbidities, or after failed conventional therapies in patients who are not candidates for surgery. PMID- 15758143 TI - Stationary arterial waves in magnetic resonance angiography. PMID- 15758144 TI - The double density technique of increased visualization of cement during vertebroplasty: a technical note. PMID- 15758145 TI - Creation of a transjugular intrahepatic portosystemic shunt with use of a preexisting portal-hepatic vein fistula as an alternative route. PMID- 15758146 TI - Long-term follow-up after endovascular stent placement for treatment of nutcracker syndrome. PMID- 15758147 TI - Suppression of hREV1 expression reduces the rate at which human ovarian carcinoma cells acquire resistance to cisplatin. AB - Replicative bypass of many DNA adducts is dependent on the interaction of hREV1 with DNA polymerase zeta and potentially with members of the Y family of DNA polymerases. To examine the role of hREV1 in the development of cisplatin (DDP) resistance, a subline (2008-shREV1-3.3) of the ovarian carcinoma cell line 2008 was isolated in which stable expression of a short hairpin RNA suppressed hREV1 expression to 20% and reduced hREV1 protein level to 43% of that found in the parental cells. The 2008-shREV1-3.3 cells were 1.5-fold more sensitive to the cytotoxic effect of DDP but less sensitive to the mutagenic effect of DDP as evidenced by a 2.6- or 2.7-fold reduction in the ability to induce clones highly resistant to 6-thioguanine or DDP itself, respectively, in the surviving population. Reduction of hREV1 did not alter the initial rate of DDP adduct removal from DNA but did impair both spontaneous and DDP-induced extra chromosomal homologous recombination, as measured by the recombination-sensitive reporter vector pBHRF. DDP induced an increase in hREV1 protein level. DDP resistance at the population level evolved 2.8-fold more slowly in the 2008 shREV1-3.3 cells than in the parental cells during repeated cycles of drug exposure. The results indicate that hREV1 functions to enhance both cell survival and the generation of drug-resistant variants in the surviving population. DDP up regulates hREV1, suggesting that it may enhance its own mutagenicity. Most importantly, hREV1 controls the rate of emergence of resistance to DDP at the population level. Thus, hREV1 is an important contributor to DDP-induced genomic instability and the subsequent emergence of resistance. PMID- 15758148 TI - Nucleotide excision repair activity varies among murine spermatogenic cell types. AB - Germ cells perform a unique and critical biological function: they propagate the DNA that will be used to direct development of the next generation. Genetic integrity of germ cell DNA is essential for producing healthy and reproductively fit offspring, and yet germ cell DNA is damaged by endogenous and exogenous agents. Nucleotide excision repair (NER) is an important mechanism for coping with a variety of DNA lesions. Little is known about NER activity in spermatogenic cells. We expected that germ cells would be more efficient at DNA repair than somatic cells, and that this efficiency may be reduced with age when the prevalence of spontaneous mutations increases. In the present study, NER was measured in defined spermatogenic cell types, including premeiotic cells (A and B type spermatogonia), meiotic cells (pachytene spermatocytes), and postmeiotic haploid cells (round spermatids) and compared with NER in keratinocytes. Global genome repair and transcription-coupled repair subpathways of NER were examined. All spermatogenic cell types from young mice displayed good repair of (6-4) pyrimidone photoproducts, although the repair rate was slower than in primary keratinocytes. In aged mice, repair of 6-4 pyrimidone photoproducts was depressed in postmeiotic cells. While repair of cyclobutane pyrimidine dimers was not detected in spermatogenic cells or in keratinocytes, the transcribed strands of active genes were repaired with greater efficiency than nontranscribed strands or inactive genes in keratinocytes and in meiotic and postmeiotic cells; spermatogonia displayed low to moderate ability to repair cyclobutane pyrimidine dimers on both DNA strands regardless of transcriptional status. Overall, the data suggest cell type-specific NER activity during murine spermatogenesis, and our results have possible implications for germ cell aging. PMID- 15758149 TI - Gonadotropin-independent proliferation of the pale type A spermatogonia in the adult rhesus monkey (Macaca mulatta). AB - The goal of the present study was to examine the relative roles of testosterone (T) and FSH in the proliferation and differentiation of pale type A (Ap) spermatogonia in the rhesus monkey (Macaca mulatta). Twenty adult male monkeys were treated with daily injections of a GnRH-receptor antagonist, acyline, to suppress endogenous gonadotropin secretion during an experiment comprising three phases. Phase 1 established a chronic hypogonadotropic state marked by a profound decrease in testicular size. During phase 2, half the monkeys were implanted with T-filled capsules, and the other half received control implants. Treatment with T produced circulating T levels of approximately 15 ng/ml and normal testicular T content. At the end of phase 2, monkeys were fitted with indwelling i.v. catheters and housed in remote sampling cages for the final phase. During phase 3, five monkeys from the T- and non-T-treated groups were stimulated with recombinant human FSH. The remaining five monkeys from each group received an infusion of vehicle. On the last day of FSH or vehicle infusion, monkeys were bilaterally castrated after receiving an i.v. bolus of bromodeoxyuridine (BrdU). The BrdU labeling of Ap spermatogonia was robust in the hypogonadotropic group and was uninfluenced by treatment with T and FSH, either alone or in combination. In contrast, both T and FSH stimulated spermatogonial differentiation, and this effect was amplified by combined treatment. We conclude that marked Ap spermatogonial proliferation occurs constitutively and in a gonadotropin independent manner and that differentiation of Ap into B spermatogonia is absolutely gonadotropin dependent and may be driven by either T or FSH. PMID- 15758150 TI - Expression of basigin, an inducer of matrix metalloproteinases, in the rat ovary. AB - The extensive tissue remodeling that occurs during follicular development, ovulatory rupture, and the formation and regression of the corpus luteum (CL) requires local degradation of the extracellular environment by matrix metalloproteinases (MMPs). This report characterizes the expression pattern of basigin (Bsg), a putative regulator of MMP induction, in the rat ovary. An induced superovulation model (eCG/hCG) was used in immature rats to evaluate Bsg expression profiles in ovaries collected during the follicular phase, the preovulatory period, and the luteal lifespan. Levels of Bsg mRNA were unchanged through follicular growth (0-48 h post-eCG) and increased during postovulatory luteinization (24 and 48 h post-hCG; P < 0.01). Bsg expression persisted into pseudopregnancy (4-8 days post-hCG) and after functional luteal regression (12 days post-hCG). The profile of Bsg expression during regression of the CL was examined using a model of induced luteolysis. Both functional and structural regression was associated with a decline in Bsg expression levels. Bsg mRNA and protein localized to the theca of preovulatory follicles (12 h post-hCG) and formative and functional CL (24 h-8 days post-hCG). Bsg expression profiles in the induced ovulation and CL regression models were similar to observations made in naturally cycling mature rats. In the cycling ovary, Bsg signaling localized to newly forming CL, the theca of preovulatory follicles, and appeared to be lower in CL from previous estrous cycles. A putative regulatory mechanism of Bsg expression was identified using an in vitro model; treatment of cultured granulosa cells with hCG significantly augmented Bsg mRNA expression levels. The processes of ovulation and luteogenesis may be facilitated by Bsg expression and its induction or regulation of the MMPs. PMID- 15758151 TI - Androgens induce relaxation of contractile activity in pregnant human myometrium at term: a nongenomic action on L-type calcium channels. AB - It has long been accepted that progesterone regulates uterine contractile activity. However, little is known about the role of androgens in female physiology, and their importance and biological function on myometrial contractility so far have received limited attention. In this work, we examined the direct effect of androgens on the contractile activity of the isolated human myometrium. Myometrial biopsies were obtained, with consent, from pregnant women undergoing elective cesarean section at term. Each androgen tested (dehydroepiandrosterone, testosterone, 5alpha- and 5beta-dihydrotestosterone, androsterone, or androstanediol) caused a concentration-dependent inhibition of spontaneous contractile activity; a relaxing effect of these androgens was also observed on the contractions induced by high potassium (KCl) solution. Interestingly, nonpregnant myometrium was also sensitive to androgen-induced relaxation. 5beta-Dihydrotestosterone (5beta-DHT) was dramatically more potent than the other androgens in inducing myometrial relaxation in all preparations. Relaxation response to androgens had very rapid time courses and was affected by neither the specific antiandrogen (flutamide) nor inhibitors of protein synthesis (cycloheximide) and transcription (actinomycin D), implying that androgens act through a nongenomic mechanism. Importantly, 5beta-DHT significantly reduced the increase in intracellular calcium concentration associated with exposure to KCl in human myometrial smooth-muscle cells loaded with Fura-2-AM. The blockade of l type calcium channels seems to be involved in the nongenomic relaxing action of androgens. These observations demonstrate that androgens may play a crucial role in maintaining pregnancy. PMID- 15758152 TI - Signaling within the master clock of the brain: localized activation of mitogen activated protein kinase by gastrin-releasing peptide. AB - The circadian clock located in the mammalian suprachiasmatic nucleus (SCN) exhibits substantial heterogeneity in both its neurochemical and functional organization, with retinal input and oscillatory timekeeping functions segregated to different regions within the nucleus. Although it is clear that photic information must be relayed from directly retinorecipient cells to the population of oscillator cells within the nucleus, the intra-SCN signal (or signals) underlying such communication has yet to be identified. Gastrin-releasing peptide (GRP), which is found within calbindin-containing retinorecipient cells and causes photic-like phase shifts when applied directly to the SCN, is a candidate molecule. Here we examine the effect of GRP on both molecular and behavioral properties of the hamster circadian system. Within 30 min a third ventricle injection of GRP produces an increase in the number of cells expressing the phosphorylated form of extracellular signal-regulated kinases 1/2 (p-ERK1/2), localized in a discrete group of SCN cells that form a cap dorsal to calbindin cells and lateral to vasopressin cells. At 1 h after the peak of p-ERK expression these cap cells express c-fos, Period1, and Period2. Pharmacological blockade of ERK phosphorylation attenuates phase shifts to GRP. These data indicate that GRP is an output signal of retinorecipient SCN cells and activates a small cluster of SCN neurons. This novel cell group likely serves as a relay or integration point for communicating photic phase-resetting information to the rhythmic cells of the SCN. These findings represent a first step in deconstructing the SCN network constituting the brain clock. PMID- 15758153 TI - Certain inhibitors of synthetic amyloid beta-peptide (Abeta) fibrillogenesis block oligomerization of natural Abeta and thereby rescue long-term potentiation. AB - Recent studies support the hypothesis that soluble oligomers of amyloid beta peptide (Abeta) rather than mature amyloid fibrils are the earliest effectors of synaptic compromise in Alzheimer's disease. We took advantage of an amyloid precursor protein-overexpressing cell line that secretes SDS-stable Abeta oligomers to search for inhibitors of the pathobiological effects of natural human Abeta oligomers. Here, we identify small molecules that inhibit formation of soluble Abeta oligomers and thus abrogate their block of long-term potentiation (LTP). Furthermore, we show that cell-derived Abeta oligomers can be separated from monomers by size exclusion chromatography under nondenaturing conditions and that the isolated, soluble oligomers, but not monomers, block LTP. The identification of small molecules that inhibit early Abeta oligomer formation and rescue LTP inhibition offers a rational approach for therapeutic intervention in Alzheimer's disease and highlights the utility of our cell-culture paradigm as a useful secondary screen for compounds designed to inhibit early steps in Abeta oligomerization under biologically relevant conditions. PMID- 15758154 TI - Mutant superoxide dismutase 1 forms aggregates in the brain mitochondrial matrix of amyotrophic lateral sclerosis mice. AB - An increasing body of evidence suggests that mitochondrial dysfunction plays an important role in the pathogenesis of familial amyotrophic lateral sclerosis associated with "gain of function" mutations in Cu/Zn superoxide dismutase 1 (SOD1). SOD1 is mostly a cytosolic protein, but a portion of SOD1 is localized in mitochondria of patients with familial amyotrophic lateral sclerosis and transgenic mouse models of the disease. Despite the finding that mutant SOD1 localizes in mitochondria, the pathogenic significance of the mitochondrial mutant SOD1 remains to be elucidated. Here, we demonstrate that both wild-type and mutant human SOD1 accumulate in brain mitochondria of transgenic mice and that SOD1 displays a very complex intramitochondrial compartmentalization. For the first time, we show that, in addition to being in the mitochondrial outer membrane and intermembrane space, SOD1 is also localized in the mitochondrial matrix. Importantly, we show that aberrant SOD1 macromolecular aggregates are formed in the matrix of brain mitochondria. This suggests that mutant SOD1 in the brain mitochondrial matrix is misfolded and prone to aggregation, which may contribute to selective neuronal degeneration. PMID- 15758155 TI - Frontal and temporal dopamine release during working memory and attention tasks in healthy humans: a positron emission tomography study using the high-affinity dopamine D2 receptor ligand [11C]FLB 457. AB - Experimental studies on animals have shown that dopamine is a key neurotransmitter in the regulation of working memory (WM) functions in the prefrontal cortex. In humans, blood flow studies show prefrontal involvement in WM functions, but direct evidence for the involvement of the dopaminergic system in WM is lacking. Using positron emission tomography with a recently developed high-affinity dopamine D2 receptor tracer, [11C]FLB 457, we explored frontal, temporal, and parietal D2 receptor availability in 12 healthy volunteers while they were performing verbal WM and sustained attention tasks. During the performance of both tasks, reduced D2 receptor availability was observed in the left ventral anterior cingulate, suggesting an attention or arousal-related increase in dopamine release during these tasks. Compared with the sustained attention task, the verbal WM task reduced D2 receptor availability in the ventrolateral frontal cortex bilaterally and in the left medial temporal structures (amygdala, hippocampus), suggesting that dopamine release in these regions might have a specific role in WM. In addition, correlation analyses indicated that increased dopamine release in the right ventrolateral frontal cortex and the left ventral anterior cingulate during the WM task was associated with faster and more stable WM performance, respectively. Our results indicate that regionally specific components of the frontotemporal dopaminergic network are functionally involved in WM performance in humans. PMID- 15758156 TI - Sequential development of electrical and chemical synaptic connections generates a specific behavioral circuit in the leech. AB - Neuronal circuits form during embryonic life, even before synapses are completely mature. Developmental changes can be quantitative (e.g., connections become stronger and more reliable) or qualitative (e.g., synapses form, are lost, or switch from electrical to chemical or from excitatory to inhibitory). To explore how these synaptic events contribute to behavioral circuits, we have studied the formation of a circuit that produces local bending (LB) behavior in leech embryos. This circuit is composed of three layers of neurons: mechanosensory neurons, interneurons, and motor neurons. The only inhibition in this circuit is in the motor neuron layer; it allows the animal to contract on one side while relaxing the opposite side. LB develops in two stages: initially touching the body wall causes circumferential indentation (CI), an embryonic behavior in which contraction takes place around the whole perimeter of the segment touched; one or 2 d later, the same touch elicits adult-like LB. Application of bicuculline methiodide in embryos capable of LB switched the behavior back into CI, indicating that the development of GABAergic connections turns CI into LB. Using voltage-sensitive dyes and electrophysiological recordings, we found that electrical synapses were present early and produced CI. Inhibition appeared later, shaping the circuit that was already connected by electrical synapses and producing the adult behavior, LB. PMID- 15758158 TI - The class B scavenger receptor CD36 mediates free radical production and tissue injury in cerebral ischemia. AB - The class B scavenger receptor CD36 is involved in the cytotoxicity associated with inflammation, but its role in the inflammatory reaction that accompanies cerebral ischemia has not been examined. In this study, we investigated whether CD36 contributes to the brain damage produced by cerebral ischemia. The middle cerebral artery was transiently occluded in wild-type mice and in mice deficient in CD36. In wild-type mice, CD36 protein expression was increased in the ischemic brain, such that it was located predominantly in cells expressing the microglia/macrophage marker CD11b. The infarct produced by middle cerebral artery occlusion was 49% smaller in CD36-null mice than in wild-type controls, an effect associated with improved neurological function. The attenuation in brain injury in CD36 nulls could not be attributed to differences in cerebral blood flow during ischemia-reperfusion. However, the increase in reactive oxygen species (ROS) produced by cerebral ischemia was markedly attenuated in CD36-null mice in the early stage after reperfusion. The data unveil a previously unrecognized role of CD36 in ischemia-induced ROS production and brain injury. Modulation of CD36 signaling may provide a new strategy for the treatment of ischemic stroke. PMID- 15758157 TI - Plasticity in primary auditory cortex of monkeys with altered vocal production. AB - Response properties of primary auditory cortical neurons in the adult common marmoset monkey (Callithrix jacchus) were modified by extensive exposure to altered vocalizations that were self-generated and rehearsed frequently. A laryngeal apparatus modification procedure permanently lowered the frequency content of the native twitter call, a complex communication vocalization consisting of a series of frequency modulation (FM) sweeps. Monkeys vocalized shortly after this procedure and maintained voicing efforts until physiological evaluation 5-15 months later. The altered twitter calls improved over time, with FM sweeps approaching but never reaching the normal spectral range. Neurons with characteristic frequencies <4.3 kHz that had been weakly activated by native twitter calls were recruited to encode self-uttered altered twitter vocalizations. These neurons showed a decrease in response magnitude and an increase in temporal dispersion of response timing to twitter call and parametric FM stimuli but a normal response profile to pure tone stimuli. Tonotopic maps in voice-modified monkeys were not distorted. These findings suggest a previously unrecognized form of cortical plasticity that is specific to higher-order processes involved in the discrimination of more complex sounds, such as species specific vocalizations. PMID- 15758159 TI - Cortical metabotropic glutamate receptors contribute to habituation of a simple odor-evoked behavior. AB - Defining the circuits that are involved in production and cessation of specific behaviors is an ultimate goal of neuroscience. Short-term behavioral habituation is the response decrement observed in many behaviors that occurs during repeated presentation of non-reinforced stimuli. Within a number of invertebrate models of short-term behavioral habituation, depression of a defined synapse has been implicated as the mechanism. However, the synaptic mechanisms of short-term behavioral habituation have not been identified within mammals. We have shown previously that a presynaptic metabotropic glutamate receptor (mGluR)-dependent depression of synapses formed by olfactory bulb afferents to the piriform (olfactory) cortex significantly contributes to adaptation of cortical odor responses. Here we show that blockade of mGluRs within the olfactory cortex of awake, behaving rats diminishes habituation of a simple odor-induced behavior, strongly implicating a central mechanism for sensory gating in olfaction. PMID- 15758160 TI - Isotropic fractionator: a simple, rapid method for the quantification of total cell and neuron numbers in the brain. AB - Stereological techniques that estimate cell numbers must be restricted to well defined structures of isotropic architecture and therefore do not apply to the whole brain or to large neural regions. We developed a novel, fast, and inexpensive method to quantify total numbers of neuronal and non-neuronal cells in the brain or any dissectable regions thereof. It consists of transforming highly anisotropic brain structures into homogeneous, isotropic suspensions of cell nuclei, which can be counted and identified immunocytochemically as neuronal or non-neuronal. Estimates of total cell, neuronal, and non-neuronal numbers can be obtained in 24 h and vary by <10% among animals. Because the estimates obtained are independent of brain volume, they can be used in comparative studies of brain-volume variation among species and in studies of phylogenesis, development, adult neurogenesis, and pathology. Applying this method to the adult rat brain, we show, for example, that it contains approximately 330 million cells, of which 200 million are neurons, and almost 70% of these are located in the cerebellum alone. Moreover, contrary to what is commonly assumed in the literature, we show that glial cells are not the majority in the rat brain. PMID- 15758161 TI - Loss of neuronal cell cycle control in ataxia-telangiectasia: a unified disease mechanism. AB - In ataxia-telangiectasia (A-T), the loss of the ataxia-telangiectasia mutated (ATM) kinase leads to a failure of cell cycle checkpoints and DNA double-strand break detection resulting in cellular radiation sensitivity and a predisposition to cancer. There is also a significant loss of neurons, in particular cerebellar granule and Purkinje cells. Mice homozygous for null alleles of atm reproduce the radiation sensitivity and high-tumor incidence of the human disease but show no significant nerve cell loss. Using immunocytochemistry, we found the re expression of cell cycle proteins in Purkinje cells and striatal neurons in both human and mouse A-T. In the mouse, we used fluorescent in situ hybridization (FISH) to document that DNA replication accompanies the reappearance of these proteins in at-risk neuronal cells. We also found the presence of significant cell cycle activity in the Purkinje cells of the atm+/- heterozygote mouse. The cell cycle events in mouse cerebellum occur primarily during the third postnatal week by both FISH and immunocytochemistry. Thus, the initiation of this ectopic cell division occurs just as the final stages of Purkinje cell development are being completed. These results suggest that loss of cell cycle control represents a common disease mechanism that underlies the defects in the affected tissues in both human and mouse diseases. PMID- 15758162 TI - A glial endogenous cannabinoid system is upregulated in the brains of macaques with simian immunodeficiency virus-induced encephalitis. AB - Recent evidence supports the notion that the endocannabinoid system may play a crucial role in neuroinflammation. We explored the changes that some elements of this system exhibit in a macaque model of encephalitis induced by simian immunodeficiency virus. Our results show that profound alterations in the distribution of specific components of the endocannabinoid system occur as a consequence of the viral infection of the brain. Specifically, expression of cannabinoid receptors of the CB2 subtype was induced in the brains of infected animals, mainly in perivascular macrophages, microglial nodules, and T lymphocytes, most likely of the CD8 subtype. In addition, the endogenous cannabinoid-degrading enzyme fatty acid amide hydrolase was overexpressed in perivascular astrocytes as well as in astrocytic processes reaching cellular infiltrates. Finally, the pattern of CB1 receptor expression was not modified in the brains of infected animals compared with that in control animals. These results resemble previous data obtained in Alzheimer's disease human tissue samples and suggest that the endocannabinoid system may participate in the development of human immunodeficiency virus-induced encephalitis, because activation of CB2 receptors expressed by immune cells is likely to reduce their antiviral response and thus could favor the CNS entry of infected monocytes. PMID- 15758163 TI - Microglial expression of the B7 family member B7 homolog 1 confers strong immune inhibition: implications for immune responses and autoimmunity in the CNS. AB - Inflammation of the CNS is usually locally limited to avoid devastating consequences. Critical players involved in this immune regulatory process are the resident immune cells of the brain, the microglia. Interactions between the growing family of B7 costimulatory ligands and their receptors are increasingly recognized as important pathways for costimulation and/or inhibition of immune responses. Human and mouse microglial cells constitutively express B7 homolog 1 (B7-H1) in vitro. However, under inflammatory conditions [presence of interferon gamma (IFN-gamma) or T-helper 1 supernatants], a significant upregulation of B7 H1 was detectable. Expression levels of B7-H1 protein on microglial cells were substantially higher compared with astrocytes or splenocytes. Coculture experiments of major histocompatibility complex class II-positive antigen presenting cells (APC) with syngeneic T cells in the presence of antigen demonstrated the functional consequences of B7-H1 expression on T-cell activation. In the presence of a neutralizing anti-B7-H1 antibody, both the production of inflammatory cytokines (IFN-gamma and interleukin-2) and the upregulation of activation markers (inducible costimulatory signal) by T cells were markedly enhanced. Interestingly, this effect was clearly more pronounced when microglial cells were used as APC, compared with astrocytes or splenocytes. Furthermore, B7-H1 was highly upregulated during the course of myelin oligodendrocyte glycoprotein-induced and proteolipid protein-induced experimental allergic encephalomyelitis in vivo. Expression was predominantly localized to areas of strongest inflammation and could be colocalized with microglial cells/macrophages as well as T cells. Together, our data propose microglial B7-H1 as an important immune inhibitory molecule capable of downregulating T-cell activation in the CNS and thus confining immunopathological damage. PMID- 15758164 TI - Organization of multisynaptic inputs from prefrontal cortex to primary motor cortex as revealed by retrograde transneuronal transport of rabies virus. AB - The organization of multisynaptic projections from the prefrontal cortex to the primary motor cortex (MI) was examined in macaque monkeys by retrograde transneuronal transport of rabies virus. In the first series of experiments, the virus was injected into the MI forelimb region, and the time-dependent distribution patterns of transsynaptic labeling were analyzed in the frontal lobe with various survivals (2-4 d). Two days after the viral injection, neuronal labeling emerged in the caudal aspects of the nonprimary motor-related areas that are known to project to the MI directly. At the same time, the motor thalamus contained labeled neurons. On the third day, cortical labeling extended into the rostral motor-related areas and, also, prearcuate area 8. Moreover, a number of labeled neurons were located in the internal pallidum and the cerebellar nuclei. At the 4 d postinjection period, neuronal labeling occurred widely in prefrontal areas as well as in the putamen and the cerebellar cortex. In the second series of experiments, the viral injection was made into the MI hindlimb region, and the distribution pattern of prefrontal labeling on the fourth day was compared with that in the forelimb-injection case. The labeled neurons in each prefrontal area were much fewer in the hindlimb-injection case than in the forelimb-injection case. Whereas ventral area 46 was most densely labeled from the forelimb region, only sparse labeling from the hindlimb region was observed in this prefrontal area. The present results suggest the importance of ventral area 46 in the cognitive control of forelimb movements. PMID- 15758165 TI - Plasmalemmal phosphatidylinositol-4,5-bisphosphate level regulates the releasable vesicle pool size in chromaffin cells. AB - During exocytosis, certain phospholipids may act as regulators of secretion. Here, we used several independent approaches to perturb the phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] level in bovine chromaffin cells to investigate how changes of plasmalemmal PI(4,5)P2 affect secretion. Membrane levels of PI(4,5)P2 were estimated by analyzing images of lawns of plasma membranes labeled with fluorescent probes specific for PI(4,5)P2. The specific PI(4,5)P2 signal was enriched in submicrometer-sized clusters. In parallel patch-clamp experiments on intact cells, we measured the secretion of catecholamines. Overexpression of phosphatidylinositol-4-phosphate-5-kinase I, or infusion of PI(4,5)P2 through the patch pipette, increased the PI(4,5)P2 level in the plasma membrane and potentiated secretion. Expression of a membrane-targeted inositol 5-phosphatase domain of synaptojanin 1 eliminated PI(4,5)P2 from the membrane and abolished secretion. An inhibitor of phosphatidylinositol-3 kinase, 2-(4-morpholinyl)-8 phenyl-4H-1-benzopyran-4-one, led to a transient increase in the PI(4,5)P2 level that was associated with a potentiation of secretion. After prolonged incubation, the level of PI(4,5)P2 decreased and secretion was inhibited. Kinetic analysis showed that changes in PI(4,5)P2 levels led to correlated changes in the size of two releasable vesicle pools, whereas their fusion kinetics remained unaffected. We conclude that during both short- and long-term manipulations of PI(4,5)P2 level secretion scales with plasma membrane PI(4,5)P2 content and that PI(4,5)P2 has an early effect on secretion by regulating the number of vesicles ready for release. PMID- 15758166 TI - Beta-amyloid-stimulated microglia induce neuron death via synergistic stimulation of tumor necrosis factor alpha and NMDA receptors. AB - Although abundant reactive microglia are found associated with beta-amyloid (Abeta) plaques in Alzheimer's disease (AD) brains, their contribution to cell loss remains speculative. A variety of studies have documented the ability of Abeta fibrils to directly stimulate microglia in vitro to assume a neurotoxic phenotype characterized by secretion of a plethora of proinflammatory molecules. Collectively, these data suggest that activated microglia play a direct role in contributing to neuron death in AD rather than simply a role in clearance after plaque deposition. Although it is clear the Abeta-stimulated microglia acutely secrete toxic oxidizing species, the identity of longer-lived neurotoxic agents remains less defined. We used Abeta-stimulated conditioned media from primary mouse microglia to identify more stable neurotoxic secretions. The NMDA receptor antagonists memantine and 2-amino-5-phosphopetanoic acid as well as soluble tumor necrosis factor alpha (TNFalpha) receptor protect neurons from microglial conditioned media-dependent death, implicating the excitatory neurotransmitter glutamate and the proinflammatory cytokine TNFalpha as effectors of microglial stimulated death. Neuron death occurs in an oxidative damage-dependent manner, requiring activity of inducible nitric oxide synthase. Toxicity results from coincident stimulation of the TNFalpha and NMDA receptors, because stimulations of either alone are insufficient to initiate cell death. These findings suggest the hypothesis that AD brains provide the appropriate microglial-mediated inflammatory environment for TNFalpha and glutamate to synergistically stimulate toxic activation of their respective signaling pathways in neurons as a contributing mechanism of cell death. PMID- 15758167 TI - Multiple interacting sites of ectopic spike electrogenesis in primary sensory neurons. AB - Ectopic discharge generated in injured afferent axons and cell somata in vivo contributes significantly to chronic neuropathic dysesthesia and pain after nerve trauma. Progress has been made toward understanding the processes responsible for this discharge using a preparation consisting of whole excised dorsal root ganglia (DRGs) with the cut nerve attached. In the in vitro preparation, however, spike activity originates in the DRG cell soma but rarely in the axon. We have now overcome this impediment to understanding the overall electrogenic processes in soma and axon, including the resulting discharge patterns, by modifying the bath medium in which recordings are made. At both sites, bursts can be triggered by subthreshold oscillations, a phasic stimulus, or spikes arising elsewhere in the neuron. In the soma, once triggered, bursts are maintained by depolarizing afterpotentials, whereas in the axon, an additional process also plays a role, delayed depolarizing potentials. This alternative process appears to be involved in "clock-like" bursting, a discharge pattern much more common in axons than somata. Ectopic spikes arise alternatively in the soma, the injured axon end (neuroma), and the region of the axonal T-junction. Discharge sequences, and even individual multiplet bursts, may be a mosaic of action potentials that originate at these alternative electrogenic sites within the neuron. Correspondingly, discharge generated at these alternative sites may interact, explaining the sometimes-complex firing patterns observed in vivo. PMID- 15758170 TI - Statistics of decision making in the leech. AB - Animals continuously decide among different behaviors, but, even in invertebrates, the mechanisms underlying choice and decision are unknown. In this article, leech spontaneous behavior was tracked and quantified for up to 12 h. We obtained a statistical characterization, in space and time domains, of the decision processes underlying selection of behavior in the leech. We found that the spatial distribution of leech position in a uniform environment is isotropic (the same in all directions), but this isotropy is broken in the presence of localized external stimuli. In the time domain, transitions among behaviors can be described by a Markov process, the structure of which (allowed states and transitions) is highly conserved across individuals. Finally, a wide range of recurrent, deterministic motifs was identified in the apparently irregular and unstructured exploratory behavior. These results provide a rigorous description of the inner dynamics that control the spontaneous and continuous flow of behavioral decisions in the leech. PMID- 15758169 TI - Fluorescent tagging of rhythmically active respiratory neurons within the pre Botzinger complex of rat medullary slice preparations. AB - Elucidation of the neuronal mechanisms underlying respiratory rhythmogenesis is a major focal point in respiratory physiology. An area of the ventrolateral medulla, the pre-Botzinger complex (preBotC), is a critical site. Attention is now focused on understanding the cellular and network properties within the preBotC that underlie this critical function. The inability to clearly identify key "rhythm-generating" neurons within the heterogeneous population of preBotC neurons has been a significant limitation. Here we report an advancement allowing precise targeting of neurons expressing neurokinin-1 receptors (NK1Rs), which are hypothesized to be essential for respiratory rhythmogenesis. The internalization of tetramethylrhodamine conjugated substance P in rhythmically active medullary slice preparations provided clear visualization of NK1R-expressing neurons for subsequent whole-cell patch-clamp recordings. Among labeled neurons, 82% were inspiratory modulated, and 25% had pacemaker properties. We propose that this approach can be used to greatly expedite progress toward understanding the neuronal processes underlying the control of breathing. PMID- 15758168 TI - A functional genetic variation of the serotonin (5-HT) transporter affects 5-HT1A receptor binding in humans. AB - In humans, 5-HT1A receptors are implicated in anxiety and depressive disorders and their treatment. However, the physiological and genetic factors controlling 5 HT1A receptor expression are undetermined in health and disease. In this study, the influence of two genetic factors on 5-HT1A receptor expression in the living human brain was assessed using the 5-HT1A-selective positron emission tomography (PET) ligand [11C]WAY 100635. After the genotyping of 140 healthy volunteers to study population frequencies of known single nucleotide polymorphisms (SNPs) in the 5-HT1A receptor gene, the influence of the common SNP [(-1018) C>G] on 5-HT1A receptor expression was examined in a group of 35 healthy individuals scanned with [11C]WAY 100635. In the PET group, we also studied the influence of a common variable number tandem repeat polymorphism [short (S) and long (L) alleles] of the 5-HT transporter (5-HTT) gene on 5-HT1A receptor density. Whereas, the 5-HT1A receptor genotype did not show any significant effects on [11C]WAY 100635 binding, 5-HT1A receptor binding potential values were lower in all brain regions in subjects with 5-HTTLPR short (SS or SL) genotypes than those with long (LL) genotypes. Although the PET groups are necessarily a small sample size for a genetic association study, our results demonstrate for the first time that a functional polymorphism in the 5-HTT gene, but not the 5-HT1A receptor gene, affects 5-HT1A receptor availability in man. The results may offer a plausible physiological mechanism underlying the association between 5-HTTLPR genotype, behavioral traits, and mood states. PMID- 15758171 TI - The slow afterhyperpolarization in hippocampal CA1 neurons covaries with spatial learning ability in aged Fisher 344 rats. AB - Rodents commonly exhibit age-related impairments in spatial learning tasks, deficits widely thought to reflect cellular or synaptic dysfunction in the hippocampus. Using whole-cell recordings, we examined the afterhyperpolarization (AHP) in CA1 pyramidal cells in hippocampal slices from young (4-6 months of age) and aged (24-26 months of age) Fisher 344 male rats that had been behaviorally characterized in the Morris water maze. The slow AHP (sAHP) recorded from learning-impaired aged rats (AI) was significantly larger than that seen in either age-matched unimpaired rats or young controls. Among aged rats, sAHP amplitude was inversely correlated with both acquisition and probe performance in the water maze. Action potential parameters among the three groups were similar, except for spike accommodation, which was more pronounced in the AI group. Intracellular application of the cAMP analog 8-CPT-cAMP suppressed the sAHP but failed to reveal any age- or performance-related differences in the medium AHP. 8 CPT-cAMP abolished the age-related difference in spike accommodation, whereas instantaneous firing frequency was unchanged. Calcium spikes were of similar amplitude in all three groups but were broader and had significantly larger tails in aged rats; these age-related changes could be mimicked in young neurons after exposure to BayK8644. The calcium spike among aged rats correlated with task acquisition in the maze but, unlike the sAHP, failed to correlate with probe performance. This is the first demonstration that sAHP amplitude covaries with spatial learning ability in aged rats, implying that CA1 excitability strongly influences certain aspects of cognitive function. Our findings also indicate that multiple processes, in addition to elevated calcium influx, conspire to induce cognitive decline during aging. PMID- 15758172 TI - Acid-sensing properties in rat gastric sensory neurons from normal and ulcerated stomach. AB - Gastric acid contributes to dyspeptic symptoms, including abdominal pain, in patients with disorders of the proximal gastrointestinal tract. To examine the molecular sensor(s) of gastric acid chemonociception, we characterized acid elicited currents in dorsal root ganglion (DRG) and nodose ganglion (NG) neurons that innervate the stomach and examined their modulation after induction of gastric ulcers. A fluorescent dye (DiI) was injected into the stomach wall to retrogradely label gastric sensory neurons. After 1-2 weeks, gastric ulcers were induced by 45 s of luminal exposure of the stomach to 60% acetic acid injected into a clamped area of the distal stomach; control animals received saline. In whole-cell voltage-clamp recordings, all gastric DRG neurons and 55% of NG neurons exhibited transient, amiloride-sensitive, acid-sensing ion-channel (ASIC) currents. In the remaining 45% of NG neurons, protons activated a slow, sustained current that was attenuated by the transient receptor potential vanilloid subtype 1 antagonist, capsazepine. The kinetics and proton sensitivity of amiloride sensitive ASIC currents differed between NG and DRG neurons. NG neurons had a lower proton sensitivity and faster kinetics, suggesting expression of specific subtypes of ASICs in the vagal and splanchnic innervation of the stomach. Effects of Zn2+ and N,N,N',N'-tetrakis-(2-pyridylmethyl)-ethylenediamine on acid-elicited currents suggest contributions of ASIC1a and ASIC2a subunits. Gastric ulcers altered the properties of acid-elicited currents by increasing pH sensitivity and current density and changing current kinetics in gastric DRG neurons. The distinct properties of NG and DRG neurons and their modulation after injury suggest differential contributions of vagal and spinal afferent neurons to chemosensation and chemonociception. PMID- 15758173 TI - A differentially autoregulated Pet-1 enhancer region is a critical target of the transcriptional cascade that governs serotonin neuron development. AB - The Pet-1 [pheochromocytoma 12 ETS (E26 transformation-specific)] gene plays a critical role in the development of serotonin (5-HT)-modulated behaviors via its control of embryonic 5-HT neuron differentiation. Pet-1 transcription is induced exclusively in 5-HT neuron postmitotic precursors before the appearance of transmitter, and its restricted expression is maintained in the adult. However, the mechanisms that direct Pet-1 expression to this single CNS neuronal cell type are unknown. Here, we show, using transgenic methods, that genomic sequences upstream, but not downstream or within the Pet-1-coding region, are sufficient for 5-HT neuron-specific transgene expression. Enhancer sequences within a 40 kb upstream fragment directed position-independent lacZ (beta-D-galactosidase) transgene expression to the developing hindbrain before the appearance of 5-HT. Moreover, virtually all of the 5-HT neurons in the adult were lacZ positive in all of the lines examined. Transgene expression in 5-HT neurons was maintained when the 40 kb fragment was truncated on its 5' end to either 12 or 1.8 kb, although position independence was then lost. Analysis of transgene expression in Pet-1 null mice indicated that Pet-1 was required to maintain the activity of the Pet-1 enhancer region in a subset of 5-HT neurons. These findings suggest that a conserved 1.8 kb region immediately flanking the Pet-1-coding region is a critical genomic target of the transcriptional cascade that governs 5-HT neuron development and provide additional evidence for 5-HT neuron heterogeneity at the genetic level. We discuss the potential application of the Pet-1 transgenes reported here to the selective genetic manipulation of 5-HT neurons. PMID- 15758174 TI - Glioblastoma-induced attraction of endogenous neural precursor cells is associated with improved survival. AB - Neural precursor cells contribute to adult neurogenesis and to limited attempts of brain repair after injury. Here we report that in a murine experimental glioblastoma model, endogenous neural precursors migrate from the subventricular zone toward the tumor and surround it. The association of endogenous precursors with syngenic tumor grafts was observed, after injecting red fluorescent protein labeled G261 cells into the caudate-putamen of transgenic mice, which express green fluorescent protein under a promoter for nestin (nestin-GFP). Fourteen days after inoculation, the nestin-GFP cells surrounded the tumors in several cell layers and expressed markers of early noncommitted and committed precursors. Nestin-GFP cells were further identified by a characteristic membrane current pattern as recorded in acute brain slices. 5-bromo-2-deoxyuridine labeling and dye tracing experiments revealed that the tumor-associated precursors originated from the subventricular zone. Moreover, in cultured explants from the subventricular zone, the neural precursors showed extensive tropism for glioblastomas. Tumor-induced endogenous precursor cell accumulation decreased with age of the recipient; this correlated with increased tumor size and shorter survival times in aged mice. Coinjection of glioblastoma cells with neural precursors improved the survival time of old mice to a level similar to that in young mice. Coculture experiments showed that neural precursors suppressed the rapid increase in tumor cell number, which is characteristic of glioblastoma, and induced glioblastoma cell apoptosis. Our results indicate that tumor cells attract endogenous precursor cells; the presence of precursor cells is antitumorigenic; and this cellular interaction decreases with aging. PMID- 15758175 TI - Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is required for the development of ischemic neuronal death. AB - Expression of Dickkopf-1 (Dkk-1), a secreted protein that negatively modulates the Wnt pathway, was induced in the hippocampus of gerbils and rats subjected to transient global cerebral ischemia as well as in cultured cortical neurons challenged with an excitotoxic pulse. In ischemic animals, the temporal and regional pattern of Dkk-1 expression correlated with the profile of neuronal death, as assessed by Nissl staining and Dkk-1 immunostaining in adjacent hippocampal sections. Treatment of ischemic animals with either Dkk-1 antisense oligonucleotides or lithium ions (which rescue the Wnt pathway acting downstream of the Dkk-1 blockade) protected vulnerable hippocampal neurons against ischemic damage. The same treatments protected cultured cortical neurons against NMDA toxicity. We conclude that induction of Dkk-1 with the ensuing inhibition of the canonical Wnt signaling pathway is required for the development of ischemic and excitotoxic neuronal death. PMID- 15758176 TI - Structural domains involved in the regulation of transmitter release by synapsins. AB - Synapsins are a family of neuron-specific phosphoproteins that regulate neurotransmitter release by associating with synaptic vesicles. Synapsins consist of a series of conserved and variable structural domains of unknown function. We performed a systematic structure-function analysis of the various domains of synapsin by assessing the actions of synapsin fragments on neurotransmitter release, presynaptic ultrastructure, and the biochemical interactions of synapsin. Injecting a peptide derived from domain A into the squid giant presynaptic terminal inhibited neurotransmitter release in a phosphorylation dependent manner. This peptide had no effect on vesicle pool size, synaptic depression, or transmitter release kinetics. In contrast, a peptide fragment from domain C reduced the number of synaptic vesicles in the periphery of the active zone and increased the rate and extent of synaptic depression. This peptide also slowed the kinetics of neurotransmitter release without affecting the number of docked vesicles. The domain C peptide, as well as another peptide from domain E that is known to have identical effects on vesicle pool size and release kinetics, both specifically interfered with the binding of synapsins to actin but not with the binding of synapsins to synaptic vesicles. This suggests that both peptides interfere with release by preventing interactions of synapsins with actin. Thus, interactions of domains C and E with the actin cytoskeleton may allow synapsins to perform two roles in regulating release, whereas domain A has an actin-independent function that regulates transmitter release in a phosphorylation-sensitive manner. PMID- 15758177 TI - NMDA receptor-dependent synaptic translocation of insulin receptor substrate p53 via protein kinase C signaling. AB - The activity-dependent remodeling of postsynaptic structure is a fundamental process underlying learning and memory. Insulin receptor substrate p53 (IRSp53), a key player in cytoskeletal dynamics, is enriched in the postsynaptic density (PSD) fraction, but its significance in synaptic functions remains unclear. We report here that IRSp53 is accumulated rapidly at the postsynaptic sites of cultured hippocampal neurons after glutamate or NMDA stimulation in an actin cytoskeleton-dependent manner. Pharmacological profiles showed that a PKC inhibitor, but not other kinase inhibitors, specifically suppressed the synaptic translocation of IRSp53 in response to NMDA, and the selective activation of PKC with phorbol ester markedly induced the synaptic translocation. Reverse transcriptase-PCR and Western blotting showed that IRSp53-S is the major isoform expressed in cultured hippocampal neurons. The synaptic targeting of IRSp53-S was found to be mediated through N-terminal coiled-coil domain and the PDZ (PSD 95/Discs large/zona occludens-1)-binding sequence at its C-terminal end and regulated by the PKC phosphorylation of its N terminus. In electrophysiological experiments, overexpression of IRSp53-S wild type and IRSp53-S mutant that is spontaneously accumulated at the postsynaptic sites enhanced the postsynaptic function as detected by an increased miniature EPSC amplitude. These data suggest that IRSp53 is involved in NMDA receptor-linked synaptic plasticity via PKC signaling. PMID- 15758178 TI - Stargazin reduces desensitization and slows deactivation of the AMPA-type glutamate receptors. AB - The AMPA-type glutamate receptors mediate the majority of the fast excitatory synaptic transmission and critically contribute to synaptic plasticity in the brain, hence the existence of numerous trafficking proteins dedicated to regulation of their synaptic delivery and turnover. Stargazin (also termed gamma2) is a member of a recently identified protein family termed transmembrane AMPA receptor regulatory proteins (TARPs). TARPs physically associate with AMPA receptors and participate in their surface delivery and anchoring at the postsynaptic membrane. Here, we report that next to its trafficking roles, stargazin may also act as a positive allosteric modulator of AMPA receptor ion channel function. Coexpression of stargazin with AMPA receptor subunits, either in Xenopus oocytes or in human embryonic kidney 293 cells, significantly reduced receptor desensitization in response to glutamate. Receptor deactivation rates were also slowed, and the recovery from desensitization was accelerated. Structurally, based on the data showing a tight correlation between desensitization and the stability of the AMPA receptor intradimer interface, we propose that binding of stargazin may stabilize the receptor conformation. Functionally, our data suggest that AMPA receptors complexed with stargazin (and possibly also with other TARPs) at the postsynaptic membrane are significantly more responsive to synaptically released glutamate compared with AMPA receptors lacking stargazin/TARP interaction. The putative existence of such two states of synaptic AMPA receptors, with and without stargazin/TARP binding, may provide a novel mechanism for regulation of excitatory synaptic strength during development and/or in synaptic plasticity in the adult brain. PMID- 15758179 TI - Canonical transient receptor potential 1 plays a role in basic fibroblast growth factor (bFGF)/FGF receptor-1-induced Ca2+ entry and embryonic rat neural stem cell proliferation. AB - Basic fibroblast growth factor (bFGF) and its major receptor FGF receptor-1 (FGFR 1) play an important role in the development of the cortex. The mechanisms underlying the mitogenic role of bFGF/FGFR-1 signaling have not been elucidated. Intracellular Ca2+ concentrations ([Ca2+]i) in proliferating cortical neuroepithelial cells are markedly dependent on Ca2+ entry (Maric et al., 2000a). The absence of voltage-dependent Ca2+ entry channels, which emerge later, indicates that other membrane mechanisms regulate [Ca2+]i during proliferation. Canonical transient receptor potential (TRPC) family channels are candidates because they are voltage independent and are expressed during CNS development (Strubing et al., 2003). Here, we investigated the involvement of TRPC1 in bFGF mediated Ca2+ entry and proliferation of embryonic rat neural stem cells (NSCs). Both TRPC1 and FGFR-1 are expressed in the embryonic rat telencephalon and coimmunoprecipitate. Quantitative fluorescence-activated cell sorting analyses of phenotyped telencephalic dissociates show that approximately 80% of NSCs are TRPC1+, proliferating, and express FGFR-1. Like NSCs profiled ex vivo, NSC derived progeny proliferating in vitro coexpress TRPC1 and FGFR1. Antisense knock down of TRPC1 significantly decreases bFGF-mediated proliferation of NSC progeny, reduces the Ca2+ entry component of the Cai2+ response to bFGF without affecting Ca2+ release from intracellular stores or 1-oleoyl-2-acetyl-sn-glycerol-induced Ca2+ entry, and significantly blocks an inward cation current evoked by bFGF in proliferating NSCs. Both Ca2+ influx evoked by bFGF and NSC proliferation are attenuated by Gd3+ and SKF96365 two antagonists of agonist-stimulated Ca2+ entry. Together, these results show that TRPC1 contributes to bFGF/FGFR-1-induced Ca2+ influx, which is involved in self-renewal of embryonic rat NSCs. PMID- 15758180 TI - Differential localization of Src homology 2 domain-containing protein tyrosine phosphatase substrate-1 and CD47 and its molecular mechanisms in cultured hippocampal neurons. AB - Polarized localization of membrane proteins to axons or dendrites is important for a variety of neuronal functions, including neurite outgrowth and synaptogenesis during neural development. Src homology 2 domain-containing protein tyrosine phosphatase (SHP) substrate-1 (SHPS-1) and its ligand cluster of differentiation 47 (CD47), both of which are members of the Ig superfamily of proteins, are thought to constitute an intercellular communication system in the CNS, although the physiological functions of this CD47-SHPS-1 system remain unknown. To provide insight into these functions, we have now examined the localization of SHPS-1 and CD47 in cultured hippocampal neurons. Endogenous SHPS 1 was detected at the surface of both axons and dendrites, whereas endogenous CD47 was localized predominantly to the surface of dendrites. Forced expression of these two proteins confirmed their distinct localizations. The extracellular regions of SHPS-1 and CD47 were responsible, at least in part, for their axonal and dendritic localizations, respectively; however, the axonal localization of SHPS-1 was not mediated by any one of the three Ig domains in its extracellular region. Overexpression of SHPS-1 and CD47 in distinct neurons resulted in marked accumulation of these proteins at sites of contact between SHPS-1-expressing axons and CD47-expressing dendrites. Such contact sites exhibited an enlarged structure but did not contain the synaptic marker protein vesicle-associated membrane protein-2. These results suggest that differential localization of SHPS 1 and CD47 at axons and dendrites generates a directional intercellular communication system that potentially contributes to regulation of synaptogenesis and the formation of neural networks. PMID- 15758181 TI - Functional maturation of the macaque's lateral geniculate nucleus. AB - Vision in infant primates is poor, but it is not known which structures in the eye or brain set the main limits to its development. We studied the visual response properties of 348 neurons recorded in the lateral geniculate nucleus (LGN) of macaque monkeys aged 1 week to adult. We measured spatial and temporal frequency tuning curves and contrast responses with drifting achromatic sinusoidal gratings. Even in animals as young as 1 week, the main visual response properties of neurons in the magnocellular (M) and parvocellular (P) divisions of the LGN were qualitatively normal, including the spatial organization of receptive fields and the characteristic response properties that differentiate M- and P-cells. At 1 and 4 weeks, spatial and temporal resolution were less than one half of adult values, whereas contrast gain and peak response rates for optimal stimuli were about two-thirds of adult values. Adult levels were reached by 24 weeks. Analysis of correlations between S-potentials representing retinal inputs and LGN cells suggested that the LGN follows retinal input as faithfully in infants as in adults, implicating retinal development as the main driving force in LGN development. Comparisons with previously published psychophysical data and ideal observer models suggest that the relatively modest changes in LGN responses during maturation impose no significant limits on visual performance. In contrast to previous studies, we conclude that these limits are set by neural development in the visual cortex, not in or peripheral to the LGN. PMID- 15758182 TI - Frontal networks for learning and executing arbitrary stimulus-response associations. AB - Flexible rule learning, a behavior with obvious adaptive value, is known to depend on an intact prefrontal cortex (PFC). One simple, yet powerful, form of such learning consists of forming arbitrary stimulus-response (S-R) associations. A variety of evidence from monkey and human studies suggests that the PFC plays an important role in both forming new S-R associations and in using learned rules to select the contextually appropriate response to a particular stimulus cue. Although monkey lesion studies more strongly implicate the ventrolateral PFC (vlPFC) in S-R learning, clinical data and neurophysiology studies have implicated both the vlPFC and the dorsolateral region (dlPFC) in associative rule learning. Previous human imaging studies of S-R learning tasks, however, have not demonstrated involvement of the dlPFC. This may be because of the design of previous imaging studies, which used few stimuli and used explicitly stated one to-one S-R mapping rules that were usually practiced before scanning. Humans learn these rules very quickly, limiting the ability of imaging techniques to capture activity related to rule acquisition. To address these issues, we performed functional magnetic resonance imaging while subjects learned by trial and error to associate sets of abstract visual stimuli with arbitrary manual responses. Successful learning of this task required discernment of a categorical type of S-R rule in a block design expected to yield sustained rule representation. Our results show that distinct components of the dorsolateral, ventrolateral, and anterior PFC, lateral premotor cortex, supplementary motor area, and the striatum are involved in learning versus executing categorical S-R rules. PMID- 15758183 TI - Learning to like: a role for human orbitofrontal cortex in conditioned reward. AB - A great deal of human behavior and motivation is based on the intrinsic emotional significance of rewarding or aversive events, as well as on the associations formed between such emotional events and concurrent environmental stimuli. Recent functional neuroimaging studies have implicated the ventral striatum, orbitofrontal cortex (OFC), and amygdala in the representation of reward values and/or in the anticipation of rewarding events. Here, we use functional magnetic resonance imaging to compare brain activation during the presentation of reward with that during presentation of (conditioned) stimuli that have been paired previously with reward. Specifically, we aimed to investigate conditioned reward in the absence of explicit reward anticipation. Twenty-two healthy volunteers were scanned while monochrome visual patterns were incidentally associated with reward or negative feedback in the context of a simple card game. In the subsequent session, visual patterns, including the conditioned stimuli, were presented without reward or negative feedback, and the affective valence of these stimuli was assessed behaviorally. The presentation of reward compared with negative feedback activated the ventral striatum and OFC. Activation in the same OFC region was observed when, in the subsequent session, subjects passively viewed the stimuli that had been paired with reward, without the administration of reward and with subjects being essentially unaware of the conditioning manipulation. These findings suggest that the OFC in humans plays an important role in the representation of both rewarding stimuli and conditioned stimuli that have acquired reward value. PMID- 15758184 TI - The scaffold protein Homer1b/c links metabotropic glutamate receptor 5 to extracellular signal-regulated protein kinase cascades in neurons. AB - Group I metabotropic glutamate receptors (mGluRs) increase cellular levels of inositol-1,4,5-triphosphate (IP3) and thereby trigger intracellular Ca2+ release. Also, group I mGluRs are organized with members of Homer scaffold proteins into multiprotein complexes involved in postreceptor signaling. In this study, we investigated the relative importance of the IP3/Ca2+ signaling and novel Homer proteins in group I mGluR-mediated activation of extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) in cultured rat striatal neurons. We found that selective activation of mGluR5, but not mGluR1, increased ERK1/2 phosphorylation. Whereas the IP3/Ca2+ cascade transmits a small portion of signals from mGluR5 to ERK1/2, the member of Homer family Homer1b/c forms a central signaling pathway linking mGluR5 to ERK1/2 in a Ca2+-independent manner. This was demonstrated by the findings that the mGluR5-mediated ERK1/2 phosphorylation was mostly reduced by a cell-permeable Tat-fusion peptide that selectively disrupted the interaction of mGluR5 with the Homer1b/c and by small interfering RNAs that selectively knocked down cellular levels of Homer1b/c proteins. Furthermore, ERK1/2, when only coactivated by both IP3/Ca2+- and Homer1b/c-dependent pathways, showed the ability to phosphorylate two transcription factors, Elk-1 and cAMP response element-binding protein, and thereby facilitated c-Fos expression. Together, we have identified two coordinated signaling pathways (a conventional IP3/Ca2+ vs a novel Homer pathway) that differentially mediate the mGluR5-ERK coupling in neurons. Both the Ca2+-dependent and -independent pathways are corequired to activate ERK1/2 to a level sufficient to achieve the mGluR5-dependent synapse-to nucleus communication imperative for the transcriptional regulation. PMID- 15758185 TI - Selective requirement of Pax6, but not Emx2, in the specification and development of several nuclei of the amygdaloid complex. AB - The amygdaloid complex is a group of nuclei that are thought to originate from multiple sites of the dorsal and ventral telencephalic neuroepithelium. The mechanisms that regulate their development are essentially unknown. We studied the role of Pax6 and Emx2, two transcription factors that regulate regional specification and growth of the telencephalon, in the morphogenesis of the amygdaloid complex. We used a set of specific marker genes that identify distinct amygdaloid nuclei to analyze Pax6/Small eye and Emx2 knock-out mutant mouse brains. We found that there is a selective requirement for Pax6, but not Emx2, in the formation a subset of nuclei within the amygdaloid complex. Specifically, structures that were not previously considered to be developmentally linked, the nucleus of the lateral olfactory tract and the lateral, basolateral, and basomedial nuclei, all appear to have a common requirement for Pax6. Together, our findings provide new insights into the origins and mechanisms underlying the development of the amygdaloid complex. PMID- 15758187 TI - US senator alleges FDA tried to prevent Adderall XR withdrawal in Canada. PMID- 15758186 TI - The nitric oxide-cGMP signaling pathway differentially regulates presynaptic structural plasticity in cone and rod cells. AB - Although abundant structural plasticity in the form of axonal retraction, neurite extension, and formation of presynaptic varicosities is displayed by photoreceptors after retinal detachment and during genetic and age-related retinal degeneration, the mechanisms involved are mostly unknown. We demonstrated recently that Ca(2+) influx through cGMP-gated channels in cones and voltage gated L-type channels in rods is required for neurite extension in vitro (Zhang and Townes-Anderson, 2002). Here, we report that the nitric oxide (NO)-cGMP signaling pathway is active in photoreceptors and that its manipulation differentially regulates the structural plasticity of cone and rod cells. The NO receptor soluble guanylyl cyclase (sGC) was detected immunocytochemically in both cone and rod cells. Stimulation of sGC increased cGMP production in retinal cultures. In cone cells, quantitative analysis showed that NO or cGMP stimulated neuritic sprouting; this stimulatory effect was dependent on both Ca2+ influx through cGMP-gated channels and phosphorylation by protein kinase G (PKG). At the highest levels of cGMP, however, cone outgrowth was no longer increased. In rod photoreceptors, NO or cGMP consistently inhibited neuritic growth in a dose dependent manner; this inhibitory effect required PKG. When NO-cGMP signaling was inhibited, changes in the neuritic development of cone and rod cells were also observed but in the opposite direction. These results expand the role of cGMP in axonal activity to adult neuritogenesis and suggest an explanation for the neurite sprouting observed in an autosomal recessive form of retinitis pigmentosa that is characterized by high cGMP levels in photoreceptor layers. PMID- 15758188 TI - Sugar rush. PMID- 15758189 TI - Early warning. PMID- 15758190 TI - Reference values for diagnostic radiology: application and impact. AB - Reference values (RVs) are recommended by the American Association of Physicists in Medicine for four radiographic projections, computed tomography, fluoroscopy, and dental radiography. RVs are used to compare radiation doses from individual pieces of radiographic equipment with doses from similar equipment assessed in national surveys. RVs recommended by the American Association of Physicists in Medicine have been developed from the Nationwide Evaluation of X-ray Trends survey performed by the state radiation protection agencies with the cooperation and support of the U.S. Food and Drug Administration, the Conference of Radiation Control Program Directors, and the American College of Radiology. The RVs selected by the American Association of Physicists in Medicine represent, approximately, the 80th percentile of the survey distributions. Consequently, equipment exceeding the RVs is using higher radiation doses than is 80% of the equipment in the surveys. Radiation doses for specific projections, with standard phantoms, should be measured annually, as recommended by the American College of Radiology. When the RVs are exceeded, the medical physicist should investigate the cause and determine, in cooperation with the responsible radiologist, whether these doses are justified or the imaging system should be optimized to reduce patient radiation doses. RVs are a useful tool for comparing patient radiation doses at institutions throughout the United States and for providing information about radiographic equipment performance. PMID- 15758191 TI - CT of the ear in Pendred syndrome. AB - PURPOSE: To prospectively determine the structural anomalies of the inner ear by using thin-section computed tomography (CT) in an extended family with Pendred syndrome. MATERIALS AND METHODS: Ethics committee approved the study, and informed consent was obtained from every patient or from parents of patients under legal age. Twelve patients (three females and nine males aged 7-47 years) with Pendred syndrome (all from the same ethnic isolate and with the same mutation in the PDS gene) were evaluated for inner-ear malformation at thin section CT. Both ears were evaluated. Presence or absence of interscalar septum between upper and middle turns of the cochlea was evaluated, and vestibule and vestibular aqueduct were examined for enlargement. Modiolus was determined to be present or absent (modiolar deficiency). CT scans were evaluated in consensus by two radiologists (M.G., J.M.G.). RESULTS: All patients had inner ear malformation on both sides. Modiolus was absent and vestibule was enlarged on both sides in all 12 patients. Interscalar septum was absent in 18 (75%) of 24 ears. In eight patients, interscalar septum was absent in both ears, whereas in two patients, it was absent on only one side. Aqueduct was enlarged in 20 (80%) of 24 ears. In nine patients, both ears had enlarged aqueducts, while in two patients, only one side was abnormal. CONCLUSION: Inner ear malformation is an invariable finding in Pendred syndrome. Modiolus deficiency and vestibular enlargement were the most consistent anomalies in this population with Pendred syndrome. PMID- 15758193 TI - Type II endoleaks after endovascular repair of abdominal aortic aneurysms: natural history. AB - PURPOSE: To retrospectively determine the natural history of type II endoleaks detected at thin-section multi-detector row computed tomographic (CT) angiography. MATERIALS AND METHODS: Neither institutional review board approval nor patient informed consent was required. Between December 1999 and December 2000, 83 patients (73 men and 10 women; mean age, 61 years; range, 55-75 years) underwent endovascular repair of an infrarenal abdominal aortic aneurysm with an endoluminal stent graft. Postprocedural abdominal CT angiography was performed every 3-12 months for the evaluation of endoleaks and the maximal sac diameter. A retrospective analysis of all postprocedural CT angiographic reports was performed until November 2003 to document the presence and development of type II endoleaks and the maximal orthogonal aneurysmal sac size. Findings at CT angiography were evaluated with regard to clinical outcomes and treatment in all patients in whom type II endoleaks were observed. The postprocedural follow-up period was 1.5-4.5 years (mean, 2.5 years). RESULTS: Twenty type II endoleaks were identified in 16 (19%) of the 83 patients. Four (20%) of the 20 endoleaks were embolized secondary to an increasing aneurysmal sac size when compared with that at preoperative CT angiography. These four leaks occurred in two patients, each with two separate endoleaks. Sixteen (80%) of the 20 endoleaks in 14 patients were managed with continued observation. In these patients, the aneurysmal sac size was stable or had decreased when compared with the size at preoperative CT angiography. Ten (62.5%) of the 16 endoleaks have sealed spontaneously during the follow-up, and six (37.5%) have persisted with stable or decreased aneurysmal sac size. None of the patients experienced aneurysmal sac rupture. CONCLUSION: Type II endoleaks with a stable or decreased aneurysmal sac size can be followed up with CT angiography secondary to the high rate of spontaneous resolution and a low risk of rupture. PMID- 15758192 TI - Unenhanced multi-detector row CT in patients suspected of having urinary stone disease: effect of section width on diagnosis. AB - PURPOSE: To assess prospectively the effect of section width in multi-detector row computed tomographic (CT) evaluation of patients with acute flank pain who are suspected of having or known to have urinary stone disease. MATERIALS AND METHODS: This study was approved by the ethics committee of the authors' university, and written informed consent was obtained from all patients. One hundred forty-seven patients (age range, 11-101 years; mean, 51.4 years +/- 18.7 [standard deviation]) suspected of having urinary stone disease underwent unenhanced multi-detector row CT. CT was performed with four detector rows, a section thickness of 1.0 mm, an effective tube current-time product of 100 mAs, and a tube voltage of 120 kVp (CT dose index, 11.4 mGy). From these data, three sets of transverse images were reconstructed with section widths of 1.5, 3.0, and 5.0 mm and approximately 50% of overlap each. Scans were evaluated in varying random orders by two radiologists for the number, size, and location of uroliths and nephroliths and for the presence of phleboliths, renal cysts, and secondary signs of obstruction. The significance of differences between the numbers of detected calcifications and the numbers of associated abnormalities on the scans obtained with varying section widths was tested with the McNemar test at a P level of less than .05. Spearman rho rank correlation coefficients were calculated to assess the correlation between the presence of uroliths and the presence of secondary signs. RESULTS: Uroliths were found in 72 of 147 (49.0%) patients, and nephroliths were found in 16 patients (10.9%). There was no significant difference between section widths of 1.5 and 3.0 mm with regard to the number of detected stones (264 uroliths and 61 nephroliths for both protocols). Transverse sections 5.0-mm wide revealed significantly fewer uroliths (n = 231; P < .001) and nephroliths (n = 47; P < .016). The final diagnosis was changed in four of 72 patients. All missed renal and ureteral stones measured less than 3 mm in diameter. CONCLUSION: Overlapping 3-mm sections are sufficient for the detection of urinary stone disease. Small calculi (<3 mm) may be missed on 5.0-mm-thick sections. PMID- 15758194 TI - Identifying, recruiting, and enrolling adolescent survivors of childhood cancer into a randomized controlled trial of health promotion: preliminary experiences in the Survivor Health and Resilience Education (SHARE) Program. AB - OBJECTIVE: To report on the identification, recruitment, and enrollment of adolescent survivors of childhood cancer into an ongoing randomized controlled trial (RCT) of health promotion. METHODS: A total of 244 adolescents were contacted by mail and telephone to assess their trial eligibility. Data were collected with respect to each adolescent's demographics and trial recruitment efforts (frequency and intensity of telephone call contact); exclusion and randomization status were tracked throughout. RESULTS: Thirty-one percent of adolescents were ultimately randomized in the trial and 69% were excluded from randomization (13% were ineligible, 33% refused to participate, 22% were unreachable or nonresponsive, that is, did not respond to trial mailings or telephone calls, and less than 1% were withdrawn prior to randomization). Among all eligible adolescents, the trial's consent rate was 49%. Adolescents excluded owing to refusal resided the farthest away from the intervention site and experienced the least amount of telephone call contact time. The primary reasons for trial refusal were lack of interest in health promotion (28%) and lack of time to participate (23%). CONCLUSIONS: Health promotion RCTs among adolescent survivors of childhood cancer may help prevent and control the onset and severity cancer-treatment-related late effects. However, trial success may be contingent upon tracing nonresponsive adolescents and reducing and eliminating barriers to participation. PMID- 15758196 TI - The organization and connections of anterior and posterior parietal cortex in titi monkeys: do New World monkeys have an area 2? AB - We used multiunit electrophysiological recording techniques to examine the topographic organization of somatosensory area 3b and cortex posterior to area 3b, including area 1 and the presumptive area 5, in the New World titi monkey, Callicebus moloch. We also examined the ipsilateral and contralateral connections of these fields, as well as those in a region of cortex that appeared to be similar to both area 7b and the anterior intraparietal area (7b/AIP) described in macaque monkeys. All data were combined with architectonic analysis to generate comprehensive reconstructions. These studies led to several observations. First, area 1 in titi monkeys is not as precisely organized in terms of topographic order and receptive field size as is area 1 in macaque monkeys and a few New World monkeys. Second, cortex caudal to area 1 in titi monkeys is dominated by the representation of the hand and forelimb, and contains neurons that are often responsive to visual stimulation as well as somatic stimulation. This organization is more like area 5 described in macaque monkeys than like area 2. Third, ipsilateral and contralateral cortical connections become more broadly distributed away from area 3b towards the posterior parietal cortex. Specifically, area 3b has a relatively restricted pattern of connectivity with adjacent somatosensory fields 3a, 1, S2 and PV; area 1 has more broadly distributed connections than area 3b; and the presumptive areas 5 and 7b/AIP have highly diverse connections, including connections with motor and premotor cortex, extrastriate visual areas, auditory areas and somatosensory areas of the lateral sulcus. Fourth, the hand representation of the presumptive area 5 has dense callosal connections. Our results, together with previous studies in other primates, suggest that anterior parietal cortex has expanded in some primate lineages, perhaps in relation to manual abilities, and that the region of cortex we term area 5 is involved in integrating somatic inputs with the motor system and across hemispheres. Such connections could form the substrate for intentional reaching, grasping and intermanual transfer of information necessary for bilateral coordination of the hands. PMID- 15758195 TI - Functional neuroanatomy of anticipatory behavior: dissociation between sensory driven and memory-driven systems. AB - The ability to anticipate predictable stimuli allows faster responses. The predictive saccade (PRED) task has been shown to quickly induce such anticipatory behavior in humans. In a PRED task subjects track a visual target jumping back and forth between fixed positions at a fixed time interval. During this task, saccade latencies drop from approximately 200 ms to <80 ms as subjects anticipate target appearance. This change in saccade latency indicates that subjects' behavior shifts from being sensory driven to being memory driven. We conducted functional magnetic resonance imaging studies with 10 healthy adults performing the PRED task using a standard block design. We compared the PRED task with a visually guided saccade (VGS) task using unpredictable targets matched for number, direction and amplitude of required saccades. Our results show greater activation during the PRED task in the prefrontal, pre-supplementary motor and anterior cingulate cortices, hippocampus, mediodorsal thalamus, striatum and cerebellum. The VGS task elicited greater activation in the cortical eye fields and occipital cortex. These results demonstrate the important dissociation between sensory and predictive neural control of similar saccadic eye movements. Anticipatory behavior induced by the PRED task required less sensory-related processing activity and was subserved by a distributed cortico-subcortical memory system including prefronto-striatal circuitry. PMID- 15758198 TI - A morphogenetic model for the development of cortical convolutions. AB - The convolutions of the mammalian cortex are one of its most intriguing characteristics. Their pattern is very distinctive for different species, and there seems to be a remarkable relationship between convolutions and the architectonic and functional regionalization of the cerebral cortex. Yet the mechanisms behind the development of convolutions and their association with the cortical regionalization are poorly understood. Here we propose a morphogenetic model for the development of cortical convolutions based on the structure of the cortex as a closed surface with glial and axonal fibres pulling radially, the fundamental mechanical properties of cortex and fibres (elasticity and plasticity), and the growth of the cortical surface. The computer simulations of this model suggest that convolutions are a natural consequence of cortical growth. The model reproduces several aspects of convolutional development, such as the relationship between cortical surface and brain volume among mammals, the period of compensation in the degree of convolution observed in gyrencephalic brains and the dependence of the degree of convolution on cortical thickness. We have also studied the effect of early cortical regionalization on the development of convolutions by introducing geometric, mechanic and growth asymmetries in the model. The morphogenetic model is thus able to reproduce the gradients in the degree of convolution, the development of primary, secondary and tertiary convolution, and the overproduction of sulci observed in animals with altered afferent cortical connections. PMID- 15758197 TI - Differential localization of protein phosphatase-1alpha, beta and gamma1 isoforms in primate prefrontal cortex. AB - Prefrontal cortical functioning depends on D1 family receptors and their complex signal transduction cascade, including protein phosphatase-1 (PP1). Three PP1 isoforms are prominent in the brain: PP1alpha, PP1beta and PP1gamma1. PP1 localization by a variety of scaffolding proteins is critical for dopamine mediated modulation of glutamatergic neurotransmission. We have quantified the subcellular distribution of each isoform in primate prefrontal cortex using immunoelectron microscopy. All three are found in spines, dendrites, axon terminals, axons and glia. However, PP1alpha and PP1gamma1 labeling is enriched in spines, whereas PP1beta label is enriched in dendrites. Using post-embedding immunogold labeling, we further examined the distribution of PP1alpha and PP1gamma1 within spines. PP1gamma1 is highly and specifically concentrated in the postsynaptic density (PSD) of these spines, while PP1alpha is enriched in the PSD but also found subjacent to the PSD in moderate amounts. Thus, PP1 isoforms are heterogeneously distributed in the cortical neuropil and within spines. These results suggest that each PP1 isoform has access to a different set of substrates and, furthermore, they demonstrate that the composition of signal transduction proteins varies in different parts of the neuron and even in different regions of a dendritic spine in the primate PFC. PMID- 15758199 TI - Organization of horizontal axons in the inferior temporal cortex and primary visual cortex of the macaque monkey. AB - We investigated the organization of horizontal connections at two distinct hierarchical levels in the ventral visual cortical pathway of the monkey, the inferior temporal (TE) and primary visual (V1) cortices. After injections of anterograde tracers into layers 2 and 3, clusters of terminals ('patches') of labeled horizontal collaterals in TE appeared at various distances up to 8 mm from the injection site, while in V1 clear patches were distributed only within 2 mm. The size and spacing of these patches in TE were larger and more irregular than those observed in V1. The labeling intensity of patches in V1 declined sharply with distance from the injection site. This tendency was less obvious in TE; a number of densely labeled patches existed at distant sites beyond weakly labeled patches. While injections into both areas resulted in an elongated pattern of patches, the anisotropy was greater in TE than in V1 for injections of a similar size. Dual tracer injections and larger-sized injections further revealed that the adjacent sites in TE had spatially distinct horizontal projections, compared to those in V1. These area-specific characteristics of the horizontal connections may contribute to the differences in visual information processing of TE and V1. PMID- 15758200 TI - White matter development during childhood and adolescence: a cross-sectional diffusion tensor imaging study. AB - Maturation of brain white matter pathways is an important factor in cognitive, behavioral, emotional and motor development during childhood and adolescence. In this study, we investigate white matter maturation as reflected by changes in anisotropy and white matter density with age. Thirty-four children and adolescents aged 6-19 years received diffusion-weighted magnetic resonance imaging scans. Among these, 30 children and adolescents also received high resolution T1-weighed anatomical scans. A linear regression model was used to correlate fractional anisotropy (FA) values with age on a voxel-by-voxel basis. Within the regions that showed significant FA changes with age, a post hoc analysis was performed to investigate white matter density changes. With increasing age, FA values increased in prefrontal regions, in the internal capsule as well as in basal ganglia and thalamic pathways, the ventral visual pathways, and the corpus callosum. The posterior limb of the internal capsule, intrathalamic connections, and the corpus callosum showed the most significant overlaps between white matter density and FA changes with age. This study demonstrates that during childhood and adolescence, white matter anisotropy changes in brain regions that are important for attention, motor skills, cognitive ability, and memory. This typical developmental trajectory may be altered in individuals with disorders of development, cognition and behavior. PMID- 15758201 TI - Sociality and the rate of molecular evolution. AB - The molecular clock does not tick at a uniform rate in all taxa but may be influenced by species characteristics. Eusocial species (those with reproductive division of labor) have been predicted to have faster rates of molecular evolution than their nonsocial relatives because of greatly reduced effective population size; if most individuals in a population are nonreproductive and only one or few queens produce all the offspring, then eusocial animals could have much lower effective population sizes than their solitary relatives, which should increase the rate of substitution of "nearly neutral" mutations. An earlier study reported faster rates in eusocial honeybees and vespid wasps but failed to correct for phylogenetic nonindependence or to distinguish between potential causes of rate variation. Because sociality has evolved independently in many different lineages, it is possible to conduct a more wide-ranging study to test the generality of the relationship. We have conducted a comparative analysis of 25 phylogenetically independent pairs of social lineages and their nonsocial relatives, including bees, wasps, ants, termites, shrimps, and mole rats, using a range of available DNA sequences (mitochondrial and nuclear DNA coding for proteins and RNAs, and nontranslated sequences). By including a wide range of social taxa, we were able to test whether there is a general influence of sociality on rates of molecular evolution and to test specific predictions of the hypothesis: (1) that social species have faster rates because they have reduced effective population sizes; (2) that mitochondrial genes would show a greater effect of sociality than nuclear genes; and (3) that rates of molecular evolution should be correlated with the degree of sociality. We find no consistent pattern in rates of molecular evolution between social and nonsocial lineages and no evidence that mitochondrial genes show faster rates in social taxa. However, we show that the most highly eusocial Hymenoptera do have faster rates than their nonsocial relatives. We also find that social parasites (that utilize the workers from related species to produce their own offspring) have faster rates than their social relatives, which is consistent with an effect of lower effective population size on rate of molecular evolution. Our results illustrate the importance of allowing for phylogenetic nonindependence when conducting investigations of determinants of variation in rate of molecular evolution. PMID- 15758202 TI - Molecular evolution of cadherin-related neuronal receptor/protocadherin(alpha) (CNR/Pcdh(alpha)) gene cluster in Mus musculus subspecies. AB - The mouse cadherin-related neuronal receptor/protocadherin (CNR/Pcdh) gene clusters are located on chromosome 18. We sequenced single-nucleotide polymorphisms (SNPs) of the CNR/Pcdh(alpha)-coding region among 12 wild-derived and four laboratory strains; these included the four major subspecies groups of Mus musculus: domesticus, musculus, castaneus, and bactrianus. We detected 883 coding SNPs (cSNPs) in the CNR/Pcdh(alpha) variable exons and three in the constant exons. Among all the cSNPs, 586 synonymous (silent) and 297 nonsynonymous (amino acid exchanged) substitutions were found; therefore, the K(a)/K(s) ratio (nonsynonymous substitutions per synonymous substitution) was 0.51. The synonymous cSNPs were relatively concentrated in the first and fifth extracellular cadherin domain-encoding regions (ECs) of CNR/Pcdh(alpha). These regions have high nucleotide homology among the CNR/Pcdh(alpha) paralogs, suggesting that gene conversion events in synonymous and homologous regions of the CNR/Pcdh(alpha) cluster are related to the generation of cSNPs. A phylogenetic analysis revealed gene conversion events in the EC1 and EC5 regions. Assuming that the common sequences between rat and mouse are ancestral, the GC content of the third codon position has increased in the EC1 and EC5 regions, although biased substitutions from GC to AT were detected in all the codon positions. In addition, nonsynonymous substitutions were extremely high (11 of 13, K(a)/K(s) ratio 5.5) in the laboratory mouse strains. The artificial environment of laboratory mice may allow positive selection for nonsynonymous amino acid variations in CNR/Pcdh(alpha) during inbreeding. In this study, we analyzed the direction of cSNP generation, and concluded that subspecies-specific nucleotide substitutions and region-restricted gene conversion events may have contributed to the generation of genetic variations in the CNR/Pcdh genes within and between species. PMID- 15758203 TI - Evaluating the performance of a successive-approximations approach to parameter optimization in maximum-likelihood phylogeny estimation. AB - Almost all studies that estimate phylogenies from DNA sequence data under the maximum-likelihood (ML) criterion employ an approximate approach. Most commonly, model parameters are estimated on some initial phylogenetic estimate derived using a rapid method (neighbor-joining or parsimony). Parameters are then held constant during a tree search, and ideally, the procedure is repeated until convergence is achieved. However, the effectiveness of this approximation has not been formally assessed, in part because doing so requires computationally intensive, full-optimization analyses. Here, we report both indirect and direct evaluations of the effectiveness of successive approximations. We obtained an indirect evaluation by comparing the results of replicate runs on real data that use random trees to provide initial parameter estimates. For six real data sets taken from the literature, all replicate iterative searches converged to the same joint estimates of topology and model parameters, suggesting that the approximation is not starting-point dependent, as long as the heuristic searches of tree space are rigorous. We conducted a more direct assessment using simulations in which we compared the accuracy of phylogenies estimated using full optimization of all model parameters on each tree evaluated to the accuracy of trees estimated via successive approximations. There is no significant difference between the accuracy of the approximation searches relative to full-optimization searches. Our results demonstrate that successive approximation is reliable and provide reassurance that this much faster approach is safe to use for ML estimation of topology. PMID- 15758204 TI - Positive and negative selection on mammalian Y chromosomes. AB - Y chromosomes are genetically degenerate in most organisms studied. The loss of genes from Y chromosomes is thought to be due to the inefficiency of purifying selection in nonrecombining regions, which leads to the accumulation of deleterious mutations via the processes of hitchhiking, background selection, and Muller's ratchet. As the severity of these processes depends on the number of functional genes linked together on the nonrecombining Y, it is not clear whether these processes are still at work on the old, gene-poor mammalian Y chromosomes. If purifying selection is indeed less efficient in the Y-linked, compared to the X-linked genes, deleterious nonsynonymous substitutions are expected to accumulate faster on the Y chromosome. However, positive selection on Y-linked genes could also increase the rate of amino acid-changing substitutions. Thus, the previous reports of an elevated nonsynonymous substitution rate in Y-linked genes are still open to interpretation. Here, we report evidence for positive selection in two out of three studied mammalian Y-linked genes, suggesting that adaptive Darwinian evolution may be common on mammalian Y chromosomes. Taking positive selection into account, we demonstrate that purifying selection is less efficient in mammalian Y-linked genes compared to their X-linked homologues, suggesting that these genes continue to degenerate. PMID- 15758206 TI - Energy constraints on the evolution of gene expression. AB - I here estimate the energy cost of changes in gene expression for several thousand genes in the yeast Saccharomyces cerevisiae. A doubling of gene expression, as it occurs in a gene duplication event, is significantly selected against for all genes for which expression data is available. It carries a median selective disadvantage of s > 10(-5), several times greater than the selection coefficient s = 1.47 x 10(-7) below which genetic drift dominates a mutant's fate. When considered separately, increases in messenger RNA expression or protein expression by more than a factor 2 also have significant energy costs for most genes. This means that the evolution of transcription and translation rates is not an evolutionarily neutral process. They are under active selection opposing them. My estimates are based on genome-scale information of gene expression in the yeast S. cerevisiae as well as information on the energy cost of biosynthesizing amino acids and nucleotides. PMID- 15758205 TI - Reduced MIC gene repertoire variation in West African chimpanzees as compared to humans. AB - The human major histocompatibility complex class I chain-related (MIC) genes are members of a multicopy family showing similarity to the classical HLA-A, HLA-B, and HLA-C genes. Only the MICA and MICB genes produce functional transcripts. In chimpanzees, however, only one MIC gene is expressed, showing an intermediate character, resulting from a deletion fusing the MICA and MICB gene segments together. The present population study illustrates that all chimpanzee haplotypes sampled possess the hybrid MICA/B gene. In contrast to the human situation this gene displays reduced allelic variation. The observed repertoire reduction of the chimpanzee MICA/B gene is in conformity with the severe repertoire condensation documented for Patr-B locus lineages, probably due to the close proximity of both genes. PMID- 15758207 TI - Accuracy of rate estimation using relaxed-clock models with a critical focus on the early metazoan radiation. AB - In recent years, a number of phylogenetic methods have been developed for estimating molecular rates and divergence dates under models that relax the molecular clock constraint by allowing rate change throughout the tree. These methods are being used with increasing frequency, but there have been few studies into their accuracy. We tested the accuracy of several relaxed-clock methods (penalized likelihood and Bayesian inference using various models of rate change) using nucleotide sequences simulated on a nine-taxon tree. When the sequences evolved with a constant rate, the methods were able to infer rates accurately, but estimates were more precise when a molecular clock was assumed. When the sequences evolved under a model of auto-correlated rate change, rates were accurately estimated using penalized likelihood and by Bayesian inference using lognormal and exponential models of rate change, while other models did not perform as well. When the sequences evolved under a model of uncorrelated rate change, only Bayesian inference using an exponential rate model performed well. Collectively, the results provide a strong recommendation for using the exponential model of rate change if a conservative approach to divergence time estimation is required. A case study is presented in which we use a simulation based approach to examine the hypothesis of elevated rates in the Cambrian period, and it is found that these high rate estimates might be an artifact of the rate estimation method. If this bias is present, then the ages of metazoan divergences would be systematically underestimated. The results of this study have implications for studies of molecular rates and divergence dates. PMID- 15758208 TI - Genome update: base skews in 200+ bacterial chromosomes. PMID- 15758209 TI - The evolution of groups of cooperating bacteria and the growth rate versus yield trade-off. PMID- 15758210 TI - Fungus or bacterium and vice versa? PMID- 15758211 TI - The structure of secondary cell wall polymers: how Gram-positive bacteria stick their cell walls together. AB - The cell wall of Gram-positive bacteria has been a subject of detailed chemical study over the past five decades. Outside the cytoplasmic membrane of these organisms the fundamental polymer is peptidoglycan (PG), which is responsible for the maintenance of cell shape and osmotic stability. In addition, typical essential cell wall polymers such as teichoic or teichuronic acids are linked to some of the peptidoglycan chains. In this review these compounds are considered as 'classical' cell wall polymers. In the course of recent investigations of bacterial cell surface layers (S-layers) a different class of 'non-classical' secondary cell wall polymers (SCWPs) has been identified, which is involved in anchoring of S-layers to the bacterial cell surface. Comparative analyses have shown considerable differences in chemical composition, overall structure and charge behaviour of these SCWPs. This review discusses the progress that has been made in understanding the structural principles of SCWPs, which may have useful applications in S-layer-based 'supramolecular construction kits' in nanobiotechnology. PMID- 15758212 TI - CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. AB - The remarkable repetitive elements called CRISPRs (clustered regularly interspaced short palindromic repeats) consist of repeats interspaced with non repetitive elements or 'spacers'. CRISPRs are present in both archaea and bacteria, in association with genes involved in DNA recombination and repair. In the Yersinia pestis genome, three such elements are found at three distinct loci, one of them being highly polymorphic. The authors have sequenced a total of 109 alleles of the three Y. pestis CRISPRs and they describe 29 new spacers, most being specific to one isolate. In nine strains of Yersinia pseudotuberculosis, 132 spacers were found, of which only three are common to Y. pestis isolates. In Y. pestis of the Orientalis biovar investigated in detail here, deletion of motifs is observed but it appears that addition of new motifs to a common ancestral element is the most frequent event. This takes place at the three different loci, although at a higher rate in one of the loci, and the addition of new motifs is polarized. Interestingly, the most recently acquired spacers were found to have a homologue at another locus in the genome, the majority of these inside an inactive prophage. This is believed to be the first time that the origin of the spacers in CRISPR elements has been explained. The CRISPR structure provides a new and robust identification tool. PMID- 15758213 TI - Mutants of Mycobacterium smegmatis unable to grow at acidic pH in the presence of the protonophore carbonyl cyanide m-chlorophenylhydrazone. AB - Mycobacterium smegmatis is able to grow and survive at acidic pH, and exhibits intracellular pH homeostasis under these conditions. In this study, the authors have identified low proton permeability of the cytoplasmic membrane, and high cytoplasmic buffering capacity, as determinants of intrinsic acid resistance of M. smegmatis. To identify genes encoding proteins involved in protecting cells from acid stress, a screening method was developed using the electrogenic protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP). CCCP was used to suppress intrinsic acid resistance of M. smegmatis. The screen involved exposing cells to pH 5.0 in the presence of CCCP, and survivors were rescued at various time intervals on solid medium at pH 7.5. Cells capable of responding to intracellular acidification (due to CCCP-induced proton equilibration) will survive longer under these conditions than acid-sensitive cells. From a total pool of 5000 transposon (Tn611) insertion mutants screened, eight acid-sensitive M. smegmatis mutants were isolated. These acid-sensitive mutants were unable to grow at pH 5.0 in the presence of 1-5 microM CCCP, a concentration not lethal to the wild-type strain mc2155. The DNA flanking the site of Tn611 was identified using marker rescue in Escherichia coli, and DNA sequencing to identify the disrupted locus. Acid-sensitive mutants of M. smegmatis were disrupted in genes involved in phosphonate/phosphite assimilation, methionine biosynthesis, the PPE multigene family, xenobiotic-response regulation and lipid biosynthesis. Several of the acid-sensitive mutants were also defective in stationary-phase survival, suggesting that overlapping stress protection systems exist in M. smegmatis. PMID- 15758214 TI - Functional specificity of Candida albicans Als3p proteins and clade specificity of ALS3 alleles discriminated by the number of copies of the tandem repeat sequence in the central domain. AB - Candida albicans strain SC5314 contains two ALS3 alleles, which differ in sequence with respect to the number of copies of the 108 bp tandem repeat sequence within the central domain of the coding region. One allele (ALS3(12)) has 12 tandem repeat copies while the other (ALS3(9)) has 9 copies. Wild-type C. albicans (ALS3(12)/ALS3(9)) and those containing various ALS3 alleles (ALS3(12)/als3Delta(9), als3Delta(12)/ALS3(9) and als3Delta(12)/als3Delta(9)) were assayed for adhesion to monolayers of cultured vascular endothelial and pharyngeal epithelial cells. These assays showed obvious adhesive function for the larger Als3p protein, compared to a minor contribution to adhesion from the smaller protein. These functional differences in strain SC5314 prompted examination of ALS3 allelic diversity across the five major genetic clades of C. albicans. This analysis focused on the number of copies of the tandem repeat sequence within the central domain of the coding region and showed a range of alleles encoding from 6 to 19 tandem repeat copies. Clades differed with respect to prevalent ALS3 alleles and allele distribution, but were similar for the mean number of tandem repeat copies per ALS3 allele. Analysis of allelic pairing showed clade differences and the tendency for C. albicans strains to encode one longer and one shorter ALS3 allele. The allelic variability observed for ALS3 and its functional consequences observed in strain SC5314 highlight the importance of understanding ALS allelic diversity in order to draw accurate conclusions about Als protein function. PMID- 15758215 TI - Functional analysis and annotation of the virulence plasmid pMUM001 from Mycobacterium ulcerans. AB - The presence of a 174 kb plasmid called pMUM001 in Mycobacterium ulcerans, the first example of a mycobacterial plasmid encoding a virulence determinant, was recently reported. Over half of pMUM001 is devoted to six genes, three of which encode giant polyketide synthases (PKS) that produce mycolactone, an unusual cytotoxic lipid produced by M. ulcerans. In this present study the remaining 75 non-PKS-associated protein-coding sequences (CDS) are analysed and it is shown that pMUM001 is a low-copy-number element with a functional ori that supports replication in Mycobacterium marinum but not in the fast-growing mycobacteria Mycobacterium smegmatis and Mycobacterium fortuitum. Sequence analyses revealed a highly mosaic plasmid gene structure that is reminiscent of other large plasmids. Insertion sequences (IS) and fragments of IS, some previously unreported, are interspersed among functional gene clusters, such as those genes involved in plasmid replication, the synthesis of mycolactone, and a potential phosphorelay signal transduction system. Among the IS present on pMUM001 were multiple copies of the high-copy-number M. ulcerans elements IS2404 and IS2606. No plasmid transfer systems were identified, suggesting that trans-acting factors are required for mobilization. The results presented here provide important insights into this unusual virulence plasmid from an emerging but neglected human pathogen. PMID- 15758216 TI - Metabolic flux analysis of Escherichia coli in glucose-limited continuous culture. I. Growth-rate-dependent metabolic efficiency at steady state. AB - The Escherichia coli K-12 strain TG1 was grown at 28 degrees C in aerobic glucose limited continuous cultures at dilution rates ranging from 0.044 to 0.415 h(-1). The rates of biomass formation, the specific rates of glucose, ammonium and oxygen uptake and the specific carbon dioxide evolution rate increased linearly with the dilution rate up to 0.3 h(-1). At dilution rates between 0.3 h(-1) and 0.4 h(-1), a strong deviation from the linear increase to lower specific oxygen uptake and carbon dioxide evolution rates occurred. The biomass formation rate and the specific glucose and ammonium uptake rates did not deviate that strongly from the linear increase up to dilution rates of 0.4 h(-1). An increasing percentage of glucose carbon flow towards biomass determined by a reactor mass balance and a decreasing specific ATP production rate concomitant with a decreasing adenylate energy charge indicated higher energetic efficiency of carbon substrate utilization at higher dilution rates. Estimation of metabolic fluxes by a stoichiometric model revealed an increasing activity of the pentose phosphate pathway and a decreasing tricarboxylic acid cycle activity with increasing dilution rates, indicative of the increased NADPH and precursor demand for anabolic purposes at the expense of ATP formation through catabolic activities. Thus, increasing growth rates first result in a more energy-efficient use of the carbon substrate for biomass production, i.e. a lower portion of the carbon substrate is channelled into the respiratory, energy-generating pathway. At dilution rates above 0.4 h(-1), close to the wash-out point, respiration rates dropped sharply and accumulation of glucose and acetic acid was observed. Energy generation through acetate formation yields less ATP compared with complete oxidation of the sugar carbon substrate, but is the result of maximized energy generation under conditions of restrictions in the tricarboxylic acid cycle or in respiratory NADH turnover. Thus, the data strongly support the conclusion that, in aerobic glucose-limited continuous cultures of E. coli TG1, two different carbon limitations occur: at low dilution rates, cell growth is limited by cell carbon supply and, at high dilution rates, by energy-carbon supply. PMID- 15758217 TI - Metabolic flux analysis of Escherichia coli in glucose-limited continuous culture. II. Dynamic response to famine and feast, activation of the methylglyoxal pathway and oscillatory behaviour. AB - The metabolic dynamics of the Escherichia coli K-12 strain TG1 to feast and famine were studied in glucose-limited steady-state cultures by up- and downshifts of the dilution rate, respectively. An uncoupling of anabolic and catabolic rates was observed upon dilution rate upshifts, apparent through immediately increased glucose uptake rates which were not accompanied by an immediate increase of the growth rate but instead resulted in the temporary excretion of methylglyoxal, D- and L-lactate, pyruvate and, after a delay, acetate. The energetic state of the cell during the transient was followed by measuring the adenylate energy charge, which increased within 2 min after the upshift and declined thereafter until a new steady-state level was reached. In the downshift experiment, the adenylate energy charge behaved inversely; no by products were formed, indicating a tight coupling of anabolism and catabolism. Both dilution rate shifts were accompanied by an instantaneous increase of cAMP, presaging the subsequent changes in metabolic pathway utilization. Intracellular key metabolites of the Embden-Meyerhof-Parnas (EMP) pathway were measured to evaluate the metabolic perturbation during the upshift. Fructose 1,6-diphosphate (FDP) and dihydroxyacetone phosphate (DHAP) increased rapidly after the upshift, while glyceraldehyde 3-phosphate decreased. It is concluded that this imbalance at the branch-point of FDP induces the methylglyoxal (MG) pathway, a low-energy yielding bypass of the lower EMP pathway, through the increasing level of DHAP. MG pathway activation after the upshift was simulated by restricting anabolic rates using a stoichiometry-based metabolic model. The metabolic model predicted that low-energy-yielding catabolic pathways are utilized preferentially in the transient after the upshift. Upon severe dilution rate upshifts, an oscillatory behaviour occurred, apparent through long-term oscillations of respiratory activity, which started when the cytotoxic compound MG reached a threshold concentration of 1.5 mg l(-1) in the medium. PMID- 15758218 TI - Influence of flavomycin on ruminal fermentation and microbial populations in sheep. AB - Flavomycin is a phosphoglycolipid antibiotic that promotes growth in ruminants. The aim of this study was to characterize the effects of flavomycin on ruminal micro-organisms and their metabolic consequences. In sheep receiving a mixed grass hay/concentrate diet, inclusion of 20 mg flavomycin day(-1) decreased ruminal ammonia and total volatile fatty acid concentrations (P<0.001), but the acetate : propionate ratio was unchanged. Ruminal pH tended to be lower with flavomycin, and ammonia-production rates of ruminal digesta from control animals measured in vitro tended to be inhibited by flavomycin. Pure-culture studies indicated that anaerobic fungi, protozoa and most bacterial species were insensitive to flavomycin. Fusobacterium necrophorum was the most sensitive species tested, along with some high-activity ammonia-producing (HAP) species. Effects on F. necrophorum in vivo were inconsistent due to large inter-animal variation. HAP numbers appeared to be decreased. Changes in the rumen bacterial community structure were assessed by using denaturing-gradient gel electrophoresis (DGGE) analysis of rumen digesta 16S rRNA. DGGE profiles differed from animal to animal, but remained consistent from day to day. The community structure changed when flavomycin was introduced. The roles of F. necrophorum and HAP species in ammonia formation and of F. necrophorum in the invasion of wall tissue are consistent with the observed effects of flavomycin on ruminal ammonia formation and, in other studies, on decreasing tissue-turnover rates. PMID- 15758219 TI - The pimFABCDE operon from Rhodopseudomonas palustris mediates dicarboxylic acid degradation and participates in anaerobic benzoate degradation. AB - Bacteria in anoxic environments typically convert aromatic compounds derived from pollutants or green plants to benzoyl-CoA, and then to the C7 dicarboxylic acid derivative 3-hydroxypimelyl-CoA. Inspection of the recently completed genome sequence of the purple nonsulfur phototroph Rhodopseudomonas palustris revealed one predicted cluster of genes for the beta-oxidation of dicarboxylic acids. These genes, annotated as pimFABCDE, are predicted to encode acyl-CoA ligase, enoyl-CoA hydratase, acyl-CoA dehydrogenase and acyl-CoA transferase enzymes, which should allow the conversion of odd-chain dicarboxylic acids to glutaryl CoA, and even-chain dicarboxylic acids to succinyl-CoA. A mutant strain that was deleted in the pim gene cluster grew at about half the rate of the wild-type parent when benzoate or pimelate was supplied as the sole carbon source. The mutant grew five times more slowly than the wild-type on the C14 dicarboxylic acid tetradecanedioate. The mutant was unimpaired in growth on the C8-fatty acid caprylate. The acyl-CoA ligase predicted to be encoded by the pimA gene was purified, and found to be active with C7-C14 dicarboxylic and fatty acids. The expression of a pimA-lacZ chromosomal gene fusion increased twofold when cells were grown in the presence of straight-chain C7-C14 dicarboxylic and fatty acids. These results suggest that the beta-oxidation enzymes encoded by the pim gene cluster are active with medium-chain-length dicarboxylic acids, including pimelate. However, the finding that the pim operon deletion mutant is still able to grow on dicarboxylic acids, albeit at a slower rate, indicates that R. palustris has additional genes that can also specify the degradation of these compounds. PMID- 15758220 TI - Dissimilation of cysteate via 3-sulfolactate sulfo-lyase and a sulfate exporter in Paracoccus pantotrophus NKNCYSA. AB - Paracoccus pantotrophus NKNCYSA utilizes (R)-cysteate (2-amino-3-sulfopropionate) as a sole source of carbon and energy for growth, with either nitrate or molecular oxygen as terminal electron acceptor, and the specific utilization rate of cysteate is about 2 mkat (kg protein)(-1). The initial degradative reaction is catalysed by an (R)-cysteate : 2-oxoglutarate aminotransferase, which yields 3 sulfopyruvate. The latter was reduced to 3-sulfolactate by an NAD-linked sulfolactate dehydrogenase [3.3 mkat (kg protein)(-1)]. The inducible desulfonation reaction was not detected initially in cell extracts. However, a strongly induced protein with subunits of 8 kDa (alpha) and 42 kDa (beta) was found and purified. The corresponding genes had similarities to those encoding altronate dehydratases, which often require iron for activity. The purified enzyme could then be shown to convert 3-sulfolactate to sulfite and pyruvate and it was termed sulfolactate sulfo-lyase (Suy). A high level of sulfite dehydrogenase was also induced during growth with cysteate, and the organism excreted sulfate. A putative regulator, OrfR, was encoded upstream of suyAB on the reverse strand. Downstream of suyAB was suyZ, which was cotranscribed with suyB. The gene, an allele of tauZ, encoded a putative membrane protein with transmembrane helices (COG2855), and is a candidate to encode the sulfate exporter needed to maintain homeostasis during desulfonation. suyAB-like genes are widespread in sequenced genomes and environmental samples where, in contrast to the current annotation, several presumably encode the desulfonation of 3 sulfolactate, a component of bacterial spores. PMID- 15758221 TI - Skew-Laplace distribution in Gram-negative bacterial axenic cultures: new insights into intrinsic cellular heterogeneity. AB - The application of flow cytometry and skew-Laplace statistical analysis to assess cellular heterogeneity in Gram-negative axenic cultures is reported. In particular, fit to the log-skew-Laplace distribution for cellular side scatter or 'granulosity' is reported, and a number of theoretical and applied issues are considered in relation to the biological significance of this fit. PMID- 15758222 TI - Statistical assessment of a laboratory method for growing biofilms. AB - Microbial biofilms have been grown in laboratories using a variety of different approaches. A laboratory biofilm reactor system, called the CDC biofilm reactor (CBR) system, has been devised for growing biofilms under moderate to high fluid shear stress. The reactor incorporates 24 removable biofilm growth surfaces (coupons) for sampling and analysing the biofilm. Following preliminary experiments to verify the utility of the CBR system for growing biofilms of several clinically relevant organisms, a standard operating procedure for growing a Pseudomonas aeruginosa biofilm was created. This paper presents the results of a rigorous, intra-laboratory, statistical evaluation of the repeatability and ruggedness of that procedure as well as the results of the experiments with clinically relevant organisms. For the statistical evaluations, the outcome of interest was the density (c.f.u. cm(-2)) of viable P. aeruginosa. Replicate experiments were conducted to assess the repeatability of the log density outcome. The mean P. aeruginosa log10 density was 7.1, independent of the coupon position within the reactor. The repeatability standard deviation of the log density based on one coupon per experiment was 0.59. Analysis of variance showed that the variability of the log density was 53 % attributable to within experiment sources and 47 % attributable to between-experiments sources. The ruggedness evaluation applied response-surface design and regression analysis techniques, similar to those often used for sensitivity analyses in other fields of science and engineering. This approach provided a quantitative description of ruggedness; specifically, the amount the log density was altered by small adjustments to four key operational factors--time allowed for initial surface colonization, temperature, nutrient concentration, and fluid shear stress on the biofilm. The small size of the regression coefficient associated with each operational factor showed that the method was rugged; that is, relatively insensitive to minor perturbations of the four factors. These results demonstrate that the CBR system is a reliable experimental tool for growing a standard biofilm in the laboratory and that it can be adapted to study several different micro-organisms. PMID- 15758223 TI - Analysis of ATPases of putative secretion operons in the thermoacidophilic archaeon Sulfolobus solfataricus. AB - Gram-negative bacteria use a wide variety of complex mechanisms to secrete proteins across their membranes or to assemble secreted proteins into surface structures. As most archaea only possess a cytoplasmic membrane surrounded by a membrane-anchored S-layer, the organization of such complexes might be significantly different from that in Gram-negative bacteria. Five proteins of Sulfolobus solfataricus, SSO0120, SSO0572, SSO2316, SSO2387 and SSO2680, which are homologous to secretion ATPases of bacterial type II, type IV secretion systems and the type IV pili assembly machinery, were identified. The operon structures of these putative secretion systems encoding gene clusters and the expression patterns of the ATPases under different growth conditions were determined, and it was established that all five putative ATPases do show a divalent cation-dependent ATPase activity at high temperature. These results show that the archaeal secretion systems are related to the bacterial secretion systems and might be powered in a similar way. PMID- 15758224 TI - Evidence for two recA genes mediating DNA repair in Bacillus megaterium. AB - Isolation and subsequent knockout of a recA-homologous gene in Bacillus megaterium DSM 319 resulted in a mutant displaying increased sensitivity to mitomycin C. However, this mutant did not exhibit UV hypersensitivity, a finding which eventually led to identification of a second functional recA gene. Evidence for recA duplicates was also obtained for two other B. megaterium strains. In agreement with potential DinR boxes located within their promoter regions, expression of both genes (recA1 and recA2) was found to be damage-inducible. Transcription from the recA2 promoter was significantly higher than that of recA1. Since a recA2 knockout could not be achieved, functional complementation studies were performed in Escherichia coli. Heterologous expression in a RecA null mutant resulted in increased survival after UV irradiation and mitomycin C treatment, proving both recA gene products to be functional in DNA repair. Thus, there is evidence for an SOS-like pathway in B. megaterium that differs from that of Bacillus subtilis. PMID- 15758225 TI - Comparison of gene expression in trap cells and vegetative hyphae of the nematophagous fungus Monacrosporium haptotylum. AB - Nematode-trapping fungi enter the parasitic stage by developing specific morphological structures called traps. The global patterns of gene expression in traps and mycelium of the fungus Monacrosporium haptotylum were compared. The trap of this fungus is a unicellular spherical structure called the knob, which develops on the apex of a hyphal branch. RNA was isolated from knobs and mycelium and hybridized to a cDNA array containing probes of 2822 EST clones of M. haptotylum. Despite the fact that the knobs and mycelium were grown in the same medium, there were substantial differences in the patterns of genes expressed in the two cell types. In total, 23.3 % (657 of 2822) of the putative genes were differentially expressed in knobs versus mycelium. Several of these genes displayed sequence similarities to genes known to be involved in regulating morphogenesis and cell polarity in fungi. Among them were several putative homologues for small GTPases, such as rho1, rac1 and ras1, and a rho GDP dissociation inhibitor (rdi1). Several homologues to genes involved in stress response, protein synthesis and protein degradation, transcription, and carbon metabolism were also differentially expressed. In the last category, a glycogen phosphorylase (gph1) gene homologue, one of the most upregulated genes in the knobs as compared to mycelium, was characterized. A number of the genes that were differentially expressed in trap cells are also known to be regulated during the development of infection structures in plant-pathogenic fungi. Among them, a gas1 (mas3) gene homologue (designated gks1), which is specifically expressed in appressoria of the rice blast fungus, was characterized. PMID- 15758226 TI - Hypoxia abolishes transience of the heat-shock response in the methylotrophic yeast Hansenula polymorpha. AB - The heat-shock response is conserved amongst practically all organisms. Almost invariably, the massive heat-shock protein (Hsp) synthesis that it induces is subsequently down-regulated, making this a transient, not a sustained, stress response. This study investigated whether the heat-shock response displays any unusual features in the methylotrophic yeast Hansenula polymorpha, since this organism exhibits the highest growth temperature (49-50 degrees C) identified to date for any yeast and grows at 47 degrees C without either thermal death or detriment to final biomass yield. Maximal levels of Hsp induction were observed with a temperature upshift of H. polymorpha from 30 degrees C to 47-49 degrees C. This heat shock induces a prolonged growth arrest, heat-shock protein synthesis being down-regulated long before growth resumes at such high temperatures. A 30 degrees C to 49 degrees C heat shock also induced thermotolerance, although H. polymorpha cells in balanced growth at 49 degrees C were intrinsically thermotolerant. Unexpectedly, the normal transience of the H. polymorpha heat shock response was suppressed completely by imposing the additional stress of hypoxia at the time of the 30 degrees C to 49 degrees C temperature upshift. Hypoxia abolishing the transience of the heat-shock response appears to operate at the level of Hsp gene transcription, since the heat-induced Hsp70 mRNA was transiently induced in a heat-shocked normoxic culture but displayed sustained induction in a culture deprived of oxygen at the time of temperature upshift. PMID- 15758227 TI - Identification of the DNA-binding site of the Rgg-like regulator LasX within the lactocin S promoter region. AB - LasX regulates the transcription of the divergent operons lasXY and lasA-W, which specify the production of lactocin S in Lactobacillus sakei L45. Using histidine tagged LasX, and a DNA fragment containing the complete intergenic lasA-lasX region, electrophoresis mobility-shift (EMSA) analyses were employed to demonstrate that LasX binds to the lasA-lasX intergenic DNA. Two direct heptanucleotide motifs directly upstream of P(lasA-W), and a third imperfect copy of this motif, overlapping the -10 element of P(lasA-W), were identified as possible LasX-binding sites. To assess the role of the direct repeats in the binding of LasX to the intergenic lasA-lasX region, binding experiments were performed using DNA probes with different combinations of the repeats, and with arbitrarily chosen repeat substitutions. The result of these experiments demonstrated that only the middle repeat was required for the binding of LasX to the las-promoter region. This observation correlated with the results of subsequent reporter-gene analyses, thereby weakening the hypothesis of the involvement of the direct repeats in LasX-mediated transcription regulation. By analysing the ability of LasX to bind successively shortened derivatives of the original intergenic fragment, a tentative 19 bp minimum LasX-binding site was identified. PMID- 15758228 TI - Influence of homologous phasins (PhaP) on PHA accumulation and regulation of their expression by the transcriptional repressor PhaR in Ralstonia eutropha H16. AB - Phasins play an important role in the formation of poly(3-hydroxybutyrate) [poly(3HB)] granules and affect their size. Recently, three homologues of the phasin protein PhaP1 were identified in Ralstonia eutropha strain H16. The functions of PhaP2, PhaP3 and PhaP4 were examined by analysis of R. eutropha H16 deletion strains (DeltaphaP1, DeltaphaP2, DeltaphaP3, DeltaphaP4, DeltaphaP12, DeltaphaP123 and DeltaphaP1234). When cells were grown under conditions permissive for poly(3HB) accumulation, the wild-type strain and all single-phasin negative mutants (DeltaphaP2, DeltaphaP3 and DeltaphaP4), with the exception of DeltaphaP1, showed similar growth and poly(3HB) accumulation behaviour, and also the size and number of the granules were identical. The single DeltaphaP1 mutant and the DeltaphaP12, DeltaphaP123 and DeltaphaP1234 mutants showed an almost identical growth behaviour; however, they accumulated poly(3HB) at a significantly lower level than wild-type and the single DeltaphaP2, DeltaphaP3 or DeltaphaP4 mutants. Gel-mobility-shift assays and DNaseI footprinting experiments demonstrated the capability of the transcriptional repressor PhaR to bind to a DNA region +36 to +46 bp downstream of the phaP3 start codon. The protected sequence exhibited high similarity to the binding sites of PhaR upstream of phaP1, which were identified recently. In contrast, PhaR did not bind to the upstream or intergenic regions of phaP2 and phaP4, thus indicating that the expression of these two phasins is regulated in a different way. Our current model for the regulation of phasins in R. eutropha strain H16 was extended and confirmed. PMID- 15758229 TI - Identification of a gene cluster encoding an arginine ATP-binding-cassette transporter in the genome of the thermophilic Gram-positive bacterium Geobacillus stearothermophilus strain DSMZ 13240. AB - A single gene cluster encoding components of a putative ATP-binding cassette (ABC) transporter for basic amino acids was identified in the incomplete genome sequence of the thermophilic Gram-positive bacterium Geobacillus stearothermophilus by blast searches. The cluster comprises three genes, and these were amplified from chromosomal DNA of G. stearothermophilus, ligated into plasmid vectors and expressed in Escherichia coli. The purified solute-binding protein (designated ArtJ) was demonstrated to bind L-arginine with high affinity (Kd=0.39+/-0.06 microM). Competition experiments revealed only partial inhibition by excess L-lysine (38 %) and L-ornithine (46 %), while no inhibition was observed with L-histidine or other amino acids tested. The membrane-associated transport complex, composed of a permease (designated ArtM) and an ATPase component (designated ArtP), was solubilized from E. coli membranes by decanoylsucrose and purified by metal-affinity chromatography. The ArtMP complex, when incorporated into liposomes formed from a crude extract of G. stearothermophilus lipids, displayed ATPase activity in the presence of ArtJ only. Addition of L-arginine further stimulated the activity twofold. ATP hydrolysis was optimal at 60 degrees C and sensitive to the specific inhibitor vanadate. Analysis of kinetic parameters revealed a maximal velocity of ATP hydrolysis of 0.71 micromol Pi min(-1) (mg protein)(-1) and a Km(ATP) of 1.59 mM. Together, these results identify the ArtJMP complex as a high-affinity arginine ABC transporter. PMID- 15758230 TI - Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin. AB - Porphyromonas gingivalis, an obligately anaerobic bacterium, is implicated as a major pathogen in the development and progression of chronic periodontitis. Although expression of several virulence factors of the bacterium has been found to be affected by environmental stress such as entrance into the stationary growth phase and heat, there is relatively little information on the mechanisms that may operate in the bacterium in response to environmental stress. In this study, a novel protein (UstA) was investigated that was initially identified following two-dimensional gel analysis. Expression of UstA was upregulated in stationary phase or by exposure to atmospheric oxygen. N-terminal sequencing and database analysis with the P. gingivalis genome sequence revealed that the UstA encoding gene (ustA) was located upstream of a homologue of the usp gene encoding the universal stress protein on the chromosome. The ustA gene appeared to be transcribed in a monocistronic fashion, as revealed by primer extension and Northern blot analysis. To elucidate the role of UstA in the bacterium, chromosomal mutants carrying a disruption of the ustA gene were constructed. The ustA mutant grew slower than the wild-type parent strain in rich medium, resulting in a lower yield in stationary phase. Furthermore, in this mutant, expression levels of the P. gingivalis homologues of superoxide dismutase, thiol peroxidase and thioredoxin were markedly higher than those in the wild-type, especially in stationary phase. The ustA mutant was more resistant to diamide, a thiol-specific oxidant, than the wild-type. In addition, the ustA mutation suppressed hypersensitivities of the oxyR mutant to diamide, metronidazole and mitomycin C. These results suggest that UstA may play a significant role in oxidative stress responses in the bacterium. PMID- 15758231 TI - The interplay of glycogen metabolism and differentiation provides an insight into the developmental biology of Streptomyces coelicolor. AB - Mycelial colonies of the developmentally complex actinomycete Streptomyces coelicolor growing on solid medium contain glycogen in two distinct locations. Phase I deposits are found in a substrate mycelium region bordering the developing aerial mycelium. Their production involves GlgBI, one of two glycogen branching enzyme isoforms. Phase II deposits occur in the upper regions of aerial hyphae, in long tip cells that are dividing, or have just divided, into unigenomic prespore compartments. Their formation involves a second branching enzyme isoform, GlgBII. To find out if the gene for the second isoform, glgBII, is regulated by any of the well-studied whiA, B, G, H or I genes needed for sporulation septation, glgBI or glgBII was disrupted in a set of whi mutants, and the glycogen phenotypes examined by transmission electron microscopy. In the whiG mutants, deposits were found throughout the aerial mycelium and the adjacent region of the substrate mycelium, but the morphology of all the deposits, i.e. whether they were in the form of granules of branched glycogen or large blobs of unbranched glycan, depended solely on GlgBI. In contrast, the whiA, B, H and I mutations had no obvious effect on the pattern of glycogen deposition, or on the spatial specificity of the branching enzyme isoforms (though phase II glycogen deposits were reduced in size and abundance in the whiA and B mutants, and increased in the whiH mutant). These results indicate that glgBII is regulated (directly or indirectly) by whiG, and not by any of the other whi genes tested, and that the aerial hyphae of a whiG mutant are atypical in being physiologically similar to the substrate hyphae from which they emerge. A new role for aerial hyphae is proposed. PMID- 15758232 TI - Identification of the outer-membrane protein PagC required for the serum resistance phenotype in Salmonella enterica serovar Choleraesuis. AB - Serum resistance is a crucial virulence factor for the development of systemic infections, including bacteraemia, by many pathogenic bacteria. Salmonella enterica serovar Choleraesuis is an important enteric pathogen that causes serious systemic infections in swine and humans. Here, it was found that, when introduced into Escherichia coli, a recombinant plasmid carrying the pagC gene from a plasmid-based genomic library of S. enterica serovar Choleraesuis conferred a high-level resistance to the bactericidal activity of pooled normal swine serum. The resistance was equal to the level conferred by rck, a gene encoding a 17 kDa outer-membrane protein which promotes the serum resistance phenotype in S. enterica serovar Typhimurium. Insertional mutagenesis of the cloned pagC gene generated a mutation that resulted in the loss of the serum resistance phenotype in E. coli. When this mutation was introduced into the chromosome of S. enterica serovar Choleraesuis by homology recombination with the wild-type allele, the resulting strain could not produce PagC, and it showed a decreased level of resistance to complement-mediated killing. The mutation could be restored by introduction of the intact pagC gene on a plasmid, but not by introduction of the point-mutated pagC gene. In addition, PagC was able to promote serum resistance in the S. enterica serovar Choleraesuis LPS mutant strain, which is highly sensitive to serum killing. Although PagC is not thought to confer serum resistance directly, these results strongly suggest that PagC is an important outer-membrane protein that plays an important role in the serum resistance of S. enterica serovar Choleraesuis. PMID- 15758233 TI - Construction of a combined physical and genetic map of the chromosome of Lactobacillus acidophilus ATCC 4356 and characterization of the rRNA operons. AB - The combination of PFGE and hybridization approaches was used to study the genome of Lactobacillus acidophilus neotype strain ATCC 4356. PFGE analysis of chromosomal DNA after digestion with each of the rare-cutting restriction enzymes I-CeuI, NotI, CspI, SmaI, ApaI and SgrAI allowed the size of the circular chromosome of L. acidophilus to be estimated at 2.061 Mbp. The physical map contained 86 restriction sites for the six enzymes employed, with intervals between the sites varying from 1 to 88 kbp (approximately 0.05-4.3 % of the chromosome). Based on the physical map, a genetic map was constructed via Southern blot analyses of L. acidophilus DNA using specific gene probes. A total of 73 probes representing key genes, including 12 rRNA (rrn) genes, were positioned on the latter map. Mapping analysis also indicated the presence of four rrn operons (rrnA-D) on the chromosome, each containing a single copy of each of the three rrn genes 16S (rrl), 23S (rrs) and 5S (rrf). Operon rrnD was inverted in orientation with respect to the others and contained a long 16S-23S intergenic spacer region with tRNAIle and tRNAAla genes, whereas the other operons contained a short spacer lacking any tRNA genes. The high-resolution physical/genetic map constructed in this study provides a platform for genomic and genetic studies of Lactobacillus species and for improving industrial and probiotic strains. PMID- 15758234 TI - The plasmids of Chlamydia trachomatis and Chlamydophila pneumoniae (N16): accurate determination of copy number and the paradoxical effect of plasmid curing agents. AB - A 7.5 kbp cryptic plasmid is found in almost all isolates of Chlamydia trachomatis. Real-time PCR assays, using TaqMan chemistry, were set up to quantify accurately both the chlamydial plasmid and the single copy, chromosomal omcB gene in the infectious, elementary bodies (EBs) of C. trachomatis L1 440. Plasmid copy number was also determined in the EBs of six other lymphogranuloma venereum (LGV) isolates (serovars L1-L3), ten trachoma isolates (serovars A-C) and nine urogenital isolates (serovars D-J). The results indicated an average plasmid copy number of 4.0+/-0.8 (mean+/-95 % confidence interval) plasmids per chromosome. During the chlamydial developmental cycle, up to 7.6 plasmids per chromosome were detected, indicating an increased plasmid copy number in the actively replicating reticulate bodies. Attempts to eliminate the plasmid from strain L1 440 using the plasmid-curing agents ethidium bromide, acridine orange or imipramine/novobiocin led to a paradoxical increase in plasmid copy number. It is speculated that the stress induced by chemical curing agents may stimulate the activity of plasmid-encoded replication (Rep) proteins. In contrast to C. trachomatis, only a single isolate of Chlamydophila pneumoniae bears a plasmid. C. pneumoniae strain N16 supports a 7.4 kbp plasmid in which ORF1, encoding one of the putative Rep proteins, is disrupted by a deletion and split into two smaller ORFs. Similar assay techniques revealed 1.3+/-0.2 plasmids per chromosome (mean+/-95 % confidence interval) in EBs of this strain. These findings are in agreement with the hypothesis that the ORF1-encoded protein is involved in, but not essential for, plasmid replication and control of copy number. PMID- 15758235 TI - Diverse roles for HspR in Campylobacter jejuni revealed by the proteome, transcriptome and phenotypic characterization of an hspR mutant. AB - Campylobacter jejuni is a leading cause of bacterial gastroenteritis in the developed world. The role of a homologue of the negative transcriptional regulatory protein HspR, which in other organisms participates in the control of the heat-shock response, was investigated. Following inactivation of hspR in C. jejuni, members of the HspR regulon were identified by DNA microarray transcript profiling. In agreement with the predicted role of HspR as a negative regulator of genes involved in the heat-shock response, it was observed that the transcript amounts of 13 genes were increased in the hspR mutant, including the chaperone genes dnaK, grpE and clpB, and a gene encoding the heat-shock regulator HrcA. Proteomic analysis also revealed increased synthesis of the heat-shock proteins DnaK, GrpE, GroEL and GroES in the absence of HspR. The altered expression of chaperones was accompanied by heat sensitivity, as the hspR mutant was unable to form colonies at 44 degrees C. Surprisingly, transcriptome analysis also revealed a group of 17 genes with lower transcript levels in the hspR mutant. Of these, eight were predicted to be involved in the formation of the flagella apparatus, and the decreased expression is likely to be responsible for the reduced motility and ability to autoagglutinate that was observed for hspR mutant cells. Electron micrographs showed that mutant cells were spiral-shaped and carried intact flagella, but were elongated compared to wild-type cells. The inactivation of hspR also reduced the ability of Campylobacter to adhere to and invade human epithelial INT-407 cells in vitro, possibly as a consequence of the reduced motility or lower expression of the flagellar export apparatus in hspR mutant cells. It was concluded that, in C. jejuni, HspR influences the expression of several genes that are likely to have an impact on the ability of the bacterium to successfully survive in food products and subsequently infect the consumer. PMID- 15758236 TI - Phase variation mediated niche adaptation during prolonged experimental murine infection with Helicobacter pylori. AB - Changes in the repeats associated with the recently redefined repertoire of 31 phase-variable genes in Helicobacter pylori were investigated following murine gastric colonization for up to one year in three unrelated H. pylori strains. Between the beginning and end of the experimental period, changes were seen in ten genes (32 %), which would alter gene expression in one or more of the three strains studied. For those genes that showed repeat length changes at the longest time points, intermediate time points showed differences between the rates of change for different functional groups of genes. Genes most likely to be associated with immediate niche fitting changed most rapidly, including phospholipase A (pldA) and LPS biosynthetic genes. Other surface proteins, which may be under adaptive immune selection, changed more slowly. Restriction modification genes showed no particular temporal pattern. The number of genes that phase varied during adaptation to the murine gastric environment correlated inversely with their relative fitness as previously determined in this murine model of colonization. This suggests a role for these genes in determining initial fitness for colonization as well as in subsequent niche adaptation. In addition, a coding tandem repeat within a phase-variable gene which does not control actual gene expression was also investigated. This repeat was found to vary in copy number during colonization. This suggests that changes in the structures encoded by tandem repeats may also play a role in altered protein functions and/or immune evasion during H. pylori colonization. PMID- 15758238 TI - The vlhA loci of Mycoplasma synoviae are confined to a restricted region of the genome. AB - Mycoplasma synoviae, a major pathogen of poultry, contains a single expressed, full-length vlhA gene encoding its haemagglutinin, and a large number of vlhA pseudogenes that can be recruited by multiple site-specific recombination events to generate chimaeric variants of the expressed gene. The position and distribution of the vlhA pseudogene regions, and their relationship with the expressed gene, have not been investigated. To determine the relationship between these regions, a physical map of the M. synoviae genome was constructed using the restriction endonucleases SmaI, I-CeuI, BsiWI, ApaI and XhoI and radiolabelled probes for rrnA, recA and tufA. A cloned fragment encoding the unique portion of the expressed vlhA gene and two PCR products containing conserved regions of the ORF 3 and ORF 6 vlhA pseudogenes were used to locate the regions containing these genes on the map. The chromosome of M. synoviae was found to be 890.4 kb and the two rRNA operons were in the same orientation. Both the expressed vlhA gene and the vlhA pseudogenes were confined to the same 114 kb region of the chromosome. These findings indicate that, unlike Mycoplasma gallisepticum, in which the vlhA genes are located in several loci around the chromosome and in which antigenic variation is generated by alternating transcription of over 40 translationally competent genes, M. synoviae has all of the vlhA sequences clustered together, suggesting that close proximity is needed to facilitate the site-specific recombinations used to generate diversity in the expressed vlhA gene. PMID- 15758237 TI - The Dps-like protein Fri of Listeria monocytogenes promotes stress tolerance and intracellular multiplication in macrophage-like cells. AB - Members of the ferritin-like Dps protein family are found in a number of bacterial species, where they demonstrate the potential to bind iron, and have been implicated in tolerance to oxidative stress. In this study of the food-borne pathogen Listeria monocytogenes, the fri gene encoding a Dps homologue was deleted, and, compared to wild-type cells, it was found that the resulting mutant was less resistant to hydrogen peroxide, and demonstrated reduced survival following long-term (7-11 days) incubation in laboratory media. In view of this, it is shown that fri gene expression is controlled by the hydrogen peroxide regulator PerR, as well as the general stress sigma factor sigmaB. When fri mutant cells were transferred to iron-limiting conditions, growth was retarded relative to wild-type cells, indicating that Fri may be required for iron storage. This notion is supported by the observation that the L. monocytogenes genome appears not to encode other ferritin-like proteins. Given the role of Fri in resistance to oxidative stress, and growth under iron-limiting conditions, the ability of the fri mutant to infect mice was examined. When injected by the intraperitoneal route, the fri mutant demonstrated a reduced capacity to proliferate in the organs of infected mice relative to the wild-type, whereas when the bacteria were supplied intravenously this effect was mitigated. In addition, the mutant was impaired in its ability to survive and grow in J774.A1 mouse macrophage cells. Thus, the data suggest that Fri contributes to the ability of L. monocytogenes to survive in environments where oxidative stress and low iron availability may impede bacterial proliferation. PMID- 15758239 TI - Genetic structure and chromosomal integration site of the cryptic prophage CP 1639 encoding Shiga toxin 1. AB - The sequence of 50 625 bp of chromosomal DNA derived from Shiga-toxin (Stx) producing Escherichia coli (STEC) O111: H- strain 1639/77 was determined. This DNA fragment contains the cryptic Stx1-encoding prophage CP-1639 and its flanking chromosomal regions. The genome of CP-1639 basically resembles that of lambdoid phages in structure, but contains three IS629 elements, one of which disrupts the gene of a tail fibre component. The prophage genome lacks parts of the recombination region including integrase and excisionase genes. Moreover, a capsid protein gene is absent. CP-1639 is closely associated with an integrase gene of an ancient integrative element. This element consists of three ORFs of unknown origin and a truncated integrase gene homologous to intA of CP4-57. By PCR analysis and sequencing, it was shown that this integrative element is present in a number of non-O157 STEC serotypes and in non-STEC strains, where it is located at the 3'-end of the chromosomal ssrA gene. Whereas in most E. coli O111: H- strains, prophages are inserted in this site, E. coli O26 strains contain the integrative element not connected to a prophage. In E. coli O103 strains, the genetic structure of this region is variable. Comparison of DNA sequences of this particular site in E. coli O157: H7 strain EDL933, E. coli O111: H- strain 1639/77 and E. coli K-12 strain MG1655 showed that the ssrA gene is associated in all cases with the presence of foreign DNA. The results of this study have shown that the cryptic prophage CP-1639 is associated with an integrative element at a particular site in the E. coli chromosome that possesses high genetic variability. PMID- 15758240 TI - Analysis of virulence plasmid gene expression defines three classes of effectors in the type III secretion system of Shigella flexneri. AB - Proteins directly involved in entry and dissemination of Shigella flexneri into epithelial cells are encoded by a virulence plasmid of 200 kb. A 30-kb region (designated the entry region) of this plasmid encodes components of a type III secretion (TTS) apparatus, substrates of this apparatus and their dedicated chaperones. During growth of bacteria in broth, expression of these genes is induced at 37 degrees C and the TTS apparatus is assembled in the bacterial envelope but is not active. Secretion is activated upon contact of bacteria with host cells and is deregulated in an ipaB mutant. The plasmid encodes four transcriptional regulators, VirF, VirB, MxiE and Orf81. VirF controls transcription of virB, whose product is required for transcription of entry region genes. MxiE, with the chaperone IpgC acting as a co-activator, controls expression of several effectors that are induced under conditions of secretion. Genes under the control of Orf81 are not known. The aim of this study was to define further the repertoires of virulence plasmid genes that are under the control of (i) the growth temperature, (ii) each of the known virulence plasmid encoded transcriptional regulators (VirF, VirB, MxiE and Orf81) and (iii) the activity of the TTS apparatus. Using a macroarray analysis, the expression profiles of 71 plasmid genes were compared in the wild-type strain grown at 37 and 30 degrees C and in virF, virB, mxiE, ipaB, ipaB mxiE and orf81 mutants grown at 37 degrees C. Many genes were found to be under the control of VirB and indirectly of VirF. No alteration of expression of any gene was detected in the orf81 mutant. Expression of 13 genes was increased in the secretion-deregulated ipaB mutant in an MxiE-dependent manner. On the basis of their expression profile, substrates of the TTS apparatus can be classified into three categories: (i) those that are controlled by VirB, (ii) those that are controlled by MxiE and (iii) those that are controlled by both VirB and MxiE. The differential regulation of expression of TTS effectors in response to the TTS apparatus activity suggests that different effectors might be required at different times following contact of bacteria with host cells. PMID- 15758241 TI - Reduced initiation frequency from oriC restores viability of a temperature sensitive Escherichia coli replisome mutant. AB - The dnaX gene of Escherichia coli encodes tau and gamma clamp loader subunits of the replisome. Cells carrying the temperature-sensitive dnaX2016 mutation were induced for the SOS response at non-permissive temperature. The SOS induction most likely resulted from extensive replication fork collapse that exceeded the cells' capacity for restart. Seven mutations in the dnaA gene that partly suppressed the dnaX2016 temperature sensitivity were isolated and characterized. Each of the mutations caused a single amino acid change in domains III and IV of the DnaA protein, where nucleotide binding and DNA binding, respectively, reside. The diversity of dnaA(Sx) mutants obtained indicated that a direct interaction between the DnaA protein and tau or gamma is unlikely and that the mechanism behind suppression is related to DnaA function. All dnaA(Sx) mutant cells were compromised for initiation of DNA replication, and contained fewer active replication forks than their wild-type counterparts. Conceivably, this led to a reduced number of replication fork collapses within each dnaX2016 dnaA(Sx) cell and prevented the SOS response. Lowered availability of wild-type DnaA protein also led to partial suppression of the dnaX2016 mutation, confirming that the dnaA(Sx) mode of suppression is indirect and results from a reduced initiation frequency at oriC. PMID- 15758242 TI - Two site-specific recombinases are implicated in phenotypic variation and competitive rhizosphere colonization in Pseudomonas fluorescens. AB - The biocontrol agent Pseudomonas fluorescens F113 undergoes phenotypic variation during rhizosphere colonization, and this variation has been related to the activity of a site-specific recombinase encoded by the sss gene. Here, it is shown that a second recombinase encoded by the xerD gene is also implicated in phenotypic variation. A putative xerD gene from this strain was cloned, and sequence analysis confirmed that it encoded a site-specific recombinase of the lambda integrase family. Mutants affected in the sss or xerD genes produced a very low quantity of phenotypic variants compared to the wild-type strain, both under prolonged cultivation in the laboratory and after rhizosphere colonization, and they were severely impaired in competitive root colonization. Overexpression of the genes encoding either recombinase resulted in a substantial increment in the production of phenotypic variants under both culture and rhizosphere colonization conditions, implying that both site-specific recombinases are involved in phenotypic variation. Overexpression of the sss gene suppressed the phenotype of a xerD mutant, but overexpression of the xerD gene had no effect on the phenotype of an sss mutant. Genetic analysis of the phenotypic variants obtained after overexpression of the genes encoding both the recombinases showed that they carried mutations in the gacA/S genes, which are necessary to produce a variety of secondary metabolites. These results indicate that the Gac system is affected by the activity of the site-specific recombinases. Transcriptional fusions of the sss and xerD genes with a promoterless lacZ gene showed that both genes have a similar expression pattern, with maximal expression during stationary phase. Although the expression of both genes was independent of diffusible compounds present in root exudates, it was induced by the plant, since bacteria attached to the root showed enhanced expression. PMID- 15758243 TI - The pel genes of the Pseudomonas aeruginosa PAK strain are involved at early and late stages of biofilm formation. AB - Pseudomonas aeruginosa is a Gram-negative bacterium associated with nosocomial infections and cystic fibrosis. Chronic bacterial infections are increasingly associated with the biofilm lifestyle in which microcolonies are embedded in an extracellular matrix. Screening procedures for identifying biofilm-deficient strains have allowed the characterization of several key determinants involved in this process. Biofilm-deficient P. aeruginosa PAK strains affected in a seven gene cluster called pel were characterized. The pel genes encode proteins with similarity to components involved in polysaccharide biogenesis, of which PelF is a putative glycosyltransferase. PelG was also identified as a putative component of the polysaccharide transporter (PST) family. The pel genes were previously identified in the P. aeruginosa PA14 strain as required for the production of a glucose-rich matrix material involved in the formation of a thick pellicle and resistant biofilm. However, in PA14, the pel mutants have no clear phenotype in the initiation phase of attachment. It was shown that pel mutations in the PAK strain had little influence on biofilm initiation but, as in PA14, appeared to generate the least robust and mature biofilms. Strikingly, by constructing pel mutants in a non-piliated P. aeruginosa PAK strain, an unexpected effect of the pel mutation in the early phase of biofilm formation was discovered, since it was observed that these mutants were severely defective in the attachment process on solid surfaces. The pel gene cluster is conserved in other Gram-negative bacteria, and mutation in a Ralstonia solanacearum pelG homologue, ragG, led to an adherence defect. PMID- 15758244 TI - Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis. AB - During Bacillus subtilis spore formation, many membrane proteins that function in spore development localize to the prespore septum and, subsequently, to the outer prespore membrane. Recently, it was shown that the cell-division-specific penicillin-binding proteins (PBPs) 1 and 2b localize to the asymmetric prespore septum. Here, the author studied the localization of other PBPs, fused to green fluorescent protein (GFP), during spore formation. Fusions to PBPs 4, 2c, 2d, 2a, 3, H, 4b, 5, 4a, 4* and X were expressed during vegetative growth, and their localization was monitored during sporulation. Of these PBPs, 2c, 2d, 4b and 4* have been implicated as having a function in sporulation. It was found that PBP2c, 2d and X changed their localization, while the other PBPs tested were not affected. The putative endopeptidase PbpX appears to spiral out in a pattern that resembles FtsZ redistribution during sporulation, but a pbpX knockout strain had no distinguishable phenotype. PBP2c and 2d localize to the prespore septum and follow the membrane during engulfment, and so are redistributed to the prespore membrane. A similar pattern was observed when GFP-PBP2c was expressed in the mother cell from a sporulation-specific promoter. This work shows that various PBPs known to function during sporulation are redistributed from the cytoplasmic membrane to the prespore. PMID- 15758245 TI - Prandial glucose regulation in the glucose triad: emerging evidence and insights. AB - While it is well established that overall glycemic control reduces the complications of diabetes, the role of fasting glycemia versus postprandial glycemia in the pathophysiology of diabetes and its complications, and the relative importance of these parameters as specific targets of therapy, remain controversial. Evidence that postprandial glucose (PPG) plays an independent, modifiable role in cardiovascular disease is accumulating, largely from epidemiological studies. A large number of epidemiological studies show that high postprandial glucose is an independent risk factor for cardiovascular disease: indeed, a more powerful risk factor than fasting glucose or HbA1c. Pathophysiological hypotheses that support these observations include the contribution of postprandial glucose to HbA1c; postprandial glucose as a surrogate marker for other cardiovascular risk factors, serum lipids and triglycerides in particular; and direct toxicity of elevated glucose levels attributed to "spikes" in glucose concentration following caloric ingestion. Early interventional data suggest that therapy targeted at postprandial glucose can have a favorable impact on cardiovascular events. While more interventional studies clearly are needed, the growing weight of evidence supports a therapeutic approach more directed at excessive prandial glycemic excursions that are characteristic of both type 1 and type 2 diabetes. PMID- 15758246 TI - Effects of nateglinide on the elevation of postprandial remnant-like particle triglyceride levels in Japanese patients with type 2 diabetes assessment by meal tolerance test. AB - To elucidate the role of early insulin response in post-prandial hyperlipidemia, we examined triglyceride (TG) and remnant-like particle triglyceride (RLP-TG) levels, using a meal tolerance test (MTT) with or without the administration of nateglinide (NAT). The MTTs were performed 2 d apart in 36 drug-naive patients with type 2 diabetes who had been hospitalized for glycemic control while receiving diet therapy. Before the second MTT, patients were treated with 90 mg NAT. Treatment with NAT was associated with a significant increase in insulin levels in the treated patients 1 h after the test meal, compared to levels in non treatment. NAT treatment was also associated with a significant decrease in the level of free fatty acids 1 and 2 h after the meal, and with a significant decrease in plasma glucose levels 1, 2, and 4 h after the meal, compared to those in non-treatment. During the first MTT with NAT non-treatment, 13 patients showed serum TG levels of 200 mg/dL or greater when measured 2 h after the meal. In these 13 patients, NAT administration produced a significant decrease in TG levels 1, 2, and 6 h after the meal, as well as a significant reduction in RLP-TG levels 1 and 2 h after the meal. NAT administration was also associated with significant reductions in area under the curve (DeltaAUC) for TG and RLP-TG. These results suggest that, in a clinical setting, the early insulin response is closely associated with both postprandial glucose and postprandial lipid metabolism in Japanese patients with type 2 diabetes. PMID- 15758247 TI - Peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonist increases plasma adiponectin levels in type 2 diabetic patients with proteinuria. AB - Adiponectin appears to be an important modulator for metabolic and vascular diseases. A case-controlled study was designed to measure plasma adiponectin levels and investigate the effects of rosiglitazone on adiponectin levels in type 2 diabetic patients with proteinuria. Sixty-four patients (mean age, 46.1+/-4.6 yr; 30 male, 34 female) and 26 healthy volunteers (mean age, 45.3+/-4.8 yr; 14 male, 12 female) were included. Patients with proteinuria were treated with 4 mg/d rosiglitazone (n=21, 10 males, 11 females) for 4 wk. Adiponectin levels in patients were significantly lower than those of controls (p<0.001). There were significant negative correlations between adiponectin concentrations and insulin levels as well as homeostasis model assessment (HOMA) index in patient's group (r=-0.538, p<0.001; r=-0.393, p=0.001, respectively). There was also a significant negative correlation between plasma adiponectin concentrations and the degree of proteinuria (r=-0.526, p=0.002). Plasma adiponectin levels in patients with proteinuria (n=31; 3.91+/-2.57 microg/mL) were significantly lower than those without proteinuria (n=33; 10.15+/-1.97 microg/mL) (p<0.001). After the treatment period, adiponectin levels significantly increased (p<0.001) and proteinuria, plasma insulin, and HOMA indexes significantly decreased in treatment group (p<0.001, p<0.001, p<0.001, respectively). The results suggest that adiponectin is inversely correlated with proteinuria and treatment with peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonist rosiglitazone both corrects proteinuria and increases the low adiponectin levels in diabetic patients. PMID- 15758248 TI - Plasma adiponectin concentration in relation to severity of coronary atherosclerosis and cardiovascular risk factors in middle-aged men. AB - Adiponectin, an adipocyte-derived protein, seems to be a link between obesity, insulin resistance, and atherosclerosis. The present study investigated the association between adiponectin and coronary artery disease in middle-aged men. MATERIAL AND METHODS: We examined 48 men (aged 40-60) with angiographically confirmed coronary atherosclerosis and 19 healthy men, matched by age, as a control group. Concentrations of glucose and lipids were estimated with enzymatic methods. Plasma level of adiponectin, total and free testosterone, estradiol, estrone, DHEA-S, and insulin were estimated with RIA commercial kits. RESULTS: Men with coronary atherosclerosis had lower plasma adiponectin level than controls (16.2+/-9.2 vs 20.5+/-6.7 microg/mL; p<0.05). However, after including BMI and waist as covariate data in ANCOVA, the difference in adiponectin levels between men with CAD and controls lost statistical significance (respectively for BMI and waist: p=0.4 and p=0.7). Moreover, although not significant, adiponectin levels decreased as a function of the number of significantly narrowed coronary arteries. In a priori comparison the lowest adiponectin plasma concentration was in men with three-vessel coronary artery disease (14.3+/-9.8 microg/mL) and the high-est in controls (20.5+/-6.8 microg/mL; p=0.09). Adiponectin plasma level correlated negatively (p<0.05) with BMI, waist, percentage of total fat, fasting insulin-resistance index (FIRI), total cholesterol and triglycerides, and positively with quantitative insulin sensitivity check index (QUICKI), HDL cholesterol, total testosterone, and total testosterone/estradiol ratio. CONCLUSIONS: Our data suggest that low plasma adiponectin level is connected with insulin resistance syndrome and atherogenic lipid profile. It seems that adiponectin plays a role in pathogenesis of coronary atherosclerosis, especially in obese and insulin-resistant subjects. PMID- 15758249 TI - The effects of in utero and lactational exposure to chloroform on postnatal growth and glucose tolerance in male Wistar rats. AB - Water chlorination results in the formation of trihalomethanes (THMs) including chloroform. In human stud-ies, fetal growth restriction has been associated with exposure to THMs during pregnancy and impaired fetal growth has been associated with an increased risk of type 2 diabetes. Therefore, the objective of this study was to determine the effect of in utero and lactational exposure to chloroform on birthweight and postnatal indicators of type 2 diabetes. Female Wistar rats were given chloroform (0 microg/L, 75 microg/L) in their drinking water for 2 wk prior to mating until parturition (in utero exposure only) or until weaning (in utero+lactational exposure). At postnatal d 1 (PND1) pups of dams exposed to chloroform had significantly higher serum glucose levels and lower insulin levels, but this effect was not due to b-cell depletion in the neonatal pancreas. Glucose homeostasis in response to a glucose challenge was not changed by chloroform treatment. Chloroform exposure did not affect birthweight; however, offspring of dams exposed to chloroform had significantly impaired postnatal growth. Although fetal and neonatal exposure to chloroform did not elicit physiological changes associated with the onset of type 2 diabetes, there were physiological changes resulting in impaired postnatal growth. PMID- 15758250 TI - Klotho protein activates the PKC pathway in the kidney and testis and suppresses 25-hydroxyvitamin D3 1alpha-hydroxylase gene expression. AB - Homozygous Klotho mutant (kl-/-) mice exhibit a variety of phenotypes resembling human aging, including arteriosclerosis, infertility, skin atrophy, osteoporosis, and short life span. Calcium abnormality, one of the phenotypes in kl-/- mice, is thought to be due to the elevated gene expression of 25-hydroxyvitamin D3 1alpha hydroxylase in the kidney. We studied 25-hydroxy-vitamin D3 1alpha-hydroxylase gene expression using a Klotho plasmid that we had previously constructed for Klotho protein production. It was found that Klotho protein medium upregulated cAMP and the PKC pathway, and suppressed 25-hydroxyvitamin D3 1alpha-hydrox-ylase in kidney cells. However, both cAMP and PKC are known to elevate 25 hydroxyvitamin D3 1alpha-hydroxylase gene expression, therefore, another unknown calcium regulation pathway using Klotho protein medium might exist. Furthermore, we found that activation of the PKC pathway by Klotho was observed only in the kidney and testis, where the Klotho gene is expressed, although activation of the cAMP pathway was observed in any kind of cell. These data suggest that calcium regulation through 25-hydroxyvitamin D3 1alpha-hydroxylase by Klotho depends on non-cAMP and a non-PKC pathway and that the Klotho protein may have different signaling pathways, depending on the Klotho gene expression in different cells and organs. PMID- 15758252 TI - Short-term effect of bezafibrate on the expression of adiponectin mRNA in the adipose tissues: a study in spontaneously type 2 diabetic rats with visceral obesity. AB - The effect of short-term bezafibrate (BF) administration over time on the expression of adiponectin mRNA in the tissues was examined in Otsuka Long Evans Tokushima Fatty (OLETF) rats. Eight-week-old rats were divided into the high-dose (100 mg/kg) BF group (n=15), the low-dose (10 mg/kg) BF group (n=15), or the control group (n=15) and followed up for 14 d. Tri-glyceride and free fatty acid levels significantly decreased in a dose-dependent manner in the high-dose BF group. The insulin levels increased with time, although they were significantly lower in the high-dose BF group on d 3 and 7 than the control group. Adiponectin levels significantly increased in the high-dose BF group. On d 14 of BF administration, the levels of VLDL and chy-lomicron were significantly lower in BF groups, and adiponectin mRNA expression in the white adipose tissue was significantly higher in the high-dose BF group. Findings from this study suggest that in type 2 diabetes with insulin resistance, hypertriglyceridemia is closely linked to adiponectin. PMID- 15758251 TI - Phytoestrogen alpha-zearalanol antagonizes oxidized LDL-induced inhibition of nitric oxide production and stimulation of endothelin-1 release in human umbilical vein endothelial cells. AB - Oxidative modification of low-density lipoprotein (LDL) leads to formation of the atherogenic molecule oxidized LDL (oxLDL), which is considered to be an important mediator for vascular endothelial dysfunction and atherosclerosis. It is speculated that reduced nitric oxide (NO) release/bioavailability and enhanced release of endothelin-1 (ET-1) may contribute to oxLDL-induced endothelial dysfunction. Estrogen may improve lipid profile and inhibit oxLDL-induced endothelial damage. However, estrogen replacement therapy has been suspended due to uncertainty in benefits versus risk (such as cancer progression) in postmenopausal women. This study was designed to evaluate the effect of a novel phytoestrogen, alpha-zearalanol (alpha-ZAL), on oxLDL-induced effect on NO and ET 1 production in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with oxLDL (50 microg/mL) for 24 h in the absence or presence of alpha ZAL (0-1000 nM), 17beta-estradiol (E2, 10 nM), or the E2 receptor antagonist ICI182780 (1 microM). Levels of NO and ET-1 were measured by spectrophotometry and enzymatic immunoassay, respectively. NOS activity was evaluated by conversion of 3H-arginine to 3H-citrulline. Protein and mRNA expression of NOS and ET-1 were measured by Western blot and RT-PCR. Our results indicated that oxLDL significantly reduced NO release and NOS activity, and enhanced ET-1 pro-duction associated with reduced NOS3 (but not NOS2) expression and enhanced ET-1 mRNA expression. All these oxLDL-induced alterations were significantly attenuated or abolished by co-incubation with alpha-ZAL or E2, both through an E2 receptor dependent mechanism. alpha-ZAL, E2, and ICI182780 had no effect on NO/ET-1 release, NOS activity, or expression of NOS and ET-1. These data suggested that the phytoestrogen alpha-ZAL, like E2, may effectively antagonize oxLDL-induced decrease in NO and increase in ET-1, which may be protective for endothelial function. PMID- 15758254 TI - Comparison of equivalence and determination of diagnostic utility of min-mod and clamp methods for insulin resistance in diabetes free subjects: a meta-analysis. AB - The gold standard for quantifying insulin sensitivity (IS) is the hyperinsulinemic-euglycemic clamp (Clamp) with a cut-off point of 5x10-2 (dL/min)/(microU/mL) or less to indicate insulin resistance. Bergman's minimal model (Min-Mod) is also being used to estimate IS, but there are doubts as to its equivalence with Clamp. The objective of the present study is to determine if Clamp and the tolbutamide and insulin techniques of Min-Mod are equivalent. Meta analysis based on a bibliographic search from 1970 until the present was made for the MeSH terms: insulin resistance, hyperglycemic-clamp, euglycemic-clamp, Min Mod, minimal model approach. Concordance was determined with both simple and intraclass correlation and Bland and Altman's concordance limits using R. Three of the 109 articles found were included. The concordance limits indicate that Clamp and Min-Mod are not equivalent, which could result in diagnostic errors if the accepted cut-off point is used for both methods. Given this lack of equivalence, a ROC analysis was performed and new diagnostic cut-off points of 2.4 and 4.6x0-2 (dL/min)/(microU/mL) for insulin and tolbutamide techniques of Min-Mod, respectively, are proposed, with adequate sensitivity, specificity, and predictive value. These values should be prospectively validated. PMID- 15758253 TI - Adrenal black adenoma associated with Cushing's syndrome. AB - We report a case of adrenal black adenoma associated with Cushing's syndrome. A 41-yr-old man presented to our hospital with a 6-yr history of severe hypertension and general fatigue, and a 1-mo history of diabetes mellitus. Physical examination disclosed cushing-oid manifestations. His serum cortisol concentrations ranged from 14.0 to 15.4 microg/dL, with an ACTH level <5 pg/mL. Urinary free cortisol level was increased (125 microg/d). Cortisol was not suppressed on the overnight 1 mg oral dexamethasone suppression test (DST), 2-d low-dose DST, and 2-d high-dose DST. Abdominal computed tomography and magnetic resonance imaging studies revealed a solid round tumor approx 3 cm in diameter, located in the left adrenal gland. Left adrenalectomy was performed; the surgical specimen revealed a black ade-noma consisting of compact cells within numerous pigments that seemed to be lipofuscin in nature. PMID- 15758258 TI - Festschrift in memorial of Harold M. Frost and Proceedings of the 34th International Sun Valley Workshop on Skeletal Tissue Biology. August 1-4, 2004. Sun Valley, Idaho, USA. PMID- 15758255 TI - Bacterial endotoxin lipopolysaccharide and reactive oxygen species inhibit Leydig cell steroidogenesis via perturbation of mitochondria. AB - Chronic inflammatory disease and acute infection are well known to inhibit gonadal steroidogenesis. Previous studies have demonstrated that immune activation in response to lipopolysaccharide (LPS) results in reductions in serum testosterone, and this is a direct effect on the Leydig cell. We hypothesize that during the early onset of LPS endotoxemia in vivo, testicular macrophages produce reactive oxygen species (ROS) leading to perturbation of Leydig cell mitochondria and an inhibition in steroidogenesis. To investigate the mechanism of LPS inhibition of Leydig cell steroidogenesis, alterations in mitochondria and markers of oxi-dative stress were assessed in vivo and in Leydig cell pri- mary culture. After a single injection of mice with LPS, serum testosterone was significantly decreased within 2 h. LPS injection of mice resulted in significant reductions in steroidogenic acute regulatory protein (StAR) and 3beta hydroxysteroid dehydogenase-Delta4-Delta5 isomerase (3beta-HSD) proteins. LPS significantly increased lipid peroxidation of Leydig cell membranes, indicating that LPS results in oxidative damage in vivo. Mitochondria in Leydig cells isolated from LPS-injected mice were disrupted and showed a marked reduction in the mitochondrial membrane potential (DeltaPsim). Similar to the effects of LPS, treatment of Leydig cells with hydrogen peroxide acutely inhibited steroidogenesis, reduced StAR and 3beta-HSD protein levels, and disrupted DeltaPsim. These results suggest that LPS acutely inhibits Leydig cell function by ROS-mediated disruption of Leydig cell mitochondria. Taken together, these results demonstrate the necessity of having respiring mitochondria with an intact DeltaPsim to facilitate StAR function and Leydig cell steroidogenesis. The acute effects of LPS demonstrate how sensitive Leydig cell mitochondrial steroidogenesis is to inflammation-induced oxidative stress. PMID- 15758257 TI - Tribute to Harold M. Frost M.D. PMID- 15758259 TI - Structural characterization of rheumatoid arthritis by MRI: applications in clinical research and in clinical practice. PMID- 15758260 TI - Imaging using osteocalcin-luciferase. PMID- 15758261 TI - Multi-modality imaging of musculoskeletal disease in small animals. PMID- 15758262 TI - Molecular (and functional) imaging of articular cartilage. PMID- 15758263 TI - Yes, you can see cartilage with X-rays; diffraction enhanced X-ray imaging for soft and hard tissues. PMID- 15758264 TI - Summary--imaging of bones and joints. PMID- 15758265 TI - Vertebrate evolution and the economics of bone and muscle. PMID- 15758266 TI - Integrins in bone recognition and metastasis. PMID- 15758267 TI - Revisiting the seed and soil theory of bone metastasis: new tools, same answer. PMID- 15758268 TI - Effect of anti-TGF-beta antibodies in syngeneic mouse models of metastasis. PMID- 15758269 TI - Breast cancer bone metastasis: molecular basis of tissue tropism. PMID- 15758270 TI - Summary--bone metastasis. PMID- 15758271 TI - Root resorption associated with orthodontic force in IL-1Beta knockout mouse. PMID- 15758272 TI - In vitro and in vivo study on osteocyte-specific mechanical signaling pathways. PMID- 15758273 TI - Anabolic agents and gene expression around the bone implant interface. PMID- 15758274 TI - Bone implant interface, osteolysis and potential therapies. PMID- 15758275 TI - Interface and bulk regions in the repair, regeneration, and replacement of articular cartilage. PMID- 15758276 TI - Biomaterial characteristics important to skeletal tissue engineering. PMID- 15758277 TI - Functional integration of tissue-engineered bone constructs. PMID- 15758278 TI - Summary-Joint regeneration using functional tissue engineering. PMID- 15758279 TI - A cure for osteoporosis? PMID- 15758280 TI - Non-invasive measurements of bone strength: promise and peril. PMID- 15758281 TI - Treatment of osteoporosis with Teriparatide rhPTH(1-34). PMID- 15758282 TI - Combination and sequential therapy with PTH and bisphosphonates. PMID- 15758283 TI - Issues in designing and testing a "cure" for osteoporosis. PMID- 15758284 TI - Summary--curing osteoporosis. PMID- 15758285 TI - Extracellular Ca2+-sensing in cartilage. PMID- 15758286 TI - Calcium-sensing receptors in bone cells. PMID- 15758287 TI - Clinical effects of calcimimetics in hyperparathyroidism. PMID- 15758288 TI - Summary--calcium receptors: potential targets for novel treatments for skeletal disease. PMID- 15758290 TI - Autometallographic tracing of zinc ions in growing bone. AB - It has previously been established that zinc (Zn) supplementation increases bone dimensions and strength in growing rats. The present study aims at describing differences in the localization of loosely bound or free zinc ions, as revealed by autometallography (AMG), that might take place in the skeleton of growing rats following alimentary zinc depletion and supplementation. Male Wistar rats, 4 weeks old, were randomly divided into three groups. The rats had free access to a semi-synthetic diet with different amounts of zinc added. Group 1 was given a zinc-free (2 mg zinc/kg) diet, group 2 a 47 mg zinc/kg diet, and group 3 a 60 mg zinc/kg diet. All animals were killed after 4 weeks. Animals from each group were transcardially perfused with a 0.1 % sodium sulphide solution according to the zinc specific Neo-Timm method causing zinc ions to be bound in AMG catalytic zinc sulphur clusters. We found clusters of zinc ions localized in the mineralizing osteoid in all groups. No immediate differences in AMG staining intensity could be observed between the groups neither in the uncalcified bone nor in the osteoblasts. However, alimentary zinc supply resulted in an increase in the height of the total growth plate in a dose-dependent manner. Zinc ions were also observed in chondrocytes throughout the whole thickness of the articular and the epiphyseal cartilage as well as in the inner layer of the synovial membrane. PMID- 15758291 TI - Adjusting for the partial volume effect in cortical bone analyses of pQCT images. AB - Quantitative analyses of computed tomography images are prone to errors due to the partial volume effect which affects objects (e.g., bones) that have a different size or are assessed with different resolution. We have developed a set of equations suitable for both modeling the partial volume effect in cortical bone and for performing the corresponding adjustment. Seven hollow cylinders and 2 cuboid phantoms were made out of Al with 1% Si. The specimens were scanned with a pQCT machine (XCT2002, Stratec Medizintechnik, Pforzheim, Germany) and analyzed with the integrated software, version 5.50. Measurements were performed at different resolutions (voxel size=0.20 to 0.75 mm), both in air and in Ringer solution, and analyses were performed at different detection thresholds. Applying the correcting equations set we could reduce the errors in cortical density by about 80%. The cortical area was assessed with a negligible error at a threshold (theta0) that is equivalent to the mean of the cortical bone density and of the background density. On choosing theta0 as the detection threshold the error in density was lowered to less than 2%. We propose to assess cortical area and cortical density in several steps, first assessing the area and density thereafter. Applying this method should be beneficial whenever "true world" values are required, or objects of different size are compared. PMID- 15758296 TI - Mechanisms of bone resorption in myeloma. PMID- 15758294 TI - The secret life of bone cells. PMID- 15758297 TI - Characterization of osteoclast precursor cells in murine bone marrow. PMID- 15758298 TI - Interactions between tumor and bone alter the phenotypes of both. PMID- 15758299 TI - Cytokine regulation of osteoclast formation and function. PMID- 15758300 TI - Bone loss in chronic inflammatory conditions. PMID- 15758301 TI - Patterns of bone loss in bed-ridden healthy young male subjects: results from the Long Term Bed Rest Study in Toulouse. PMID- 15758303 TI - Introduction--Mechanisms of fracture healing and pharmacologic control. PMID- 15758304 TI - Developmental aspects of fracture healing and the use of pharmacological agents to alter healing. AB - Fracture healing is a specialized postnatal repair process that recapitulates many aspects of embryological skeletal development. While many of the molecular mechanisms that control cellular differentiation and growth during embryogenesis recur during fracture healing, these processes take place in a postnatal environment that is unique and distinct from those which exist during embryogenesis. A number of the central biological processes that are believed to be crucial in the embryonic differentiation and growth of skeletal tissues and play a functional role in fracture healing are reviewed. The functional modification of these various developmental processes of fracture healing is discussed in the context of how different pharmacological agents might alter fracture healing. PMID- 15758305 TI - VEGF and bone. PMID- 15758306 TI - Pharmacologic modulators of fracture healing: the role of cyclooxygenase inhibition. PMID- 15758307 TI - Fracture healing with an EP-2 agonist. PMID- 15758308 TI - Fracture healing with anti-resorptive agents. PMID- 15758312 TI - Tendon structure and response to changing mechanical load. PMID- 15758309 TI - Injectable rhBMP-2/CPM paste for closed fracture and minimally invasive orthopaedic repairs. PMID- 15758313 TI - Tendon micromechanics and research methods in tendinopathy. PMID- 15758314 TI - Recent progress in intrasynovial flexor tendon repair and rehabilitation. PMID- 15758316 TI - Bone growth and exercise studies: The complications of maturation. PMID- 15758317 TI - Role of physical activity on bone mineral content in young children. PMID- 15758318 TI - Lessons learned from school-based intervention trials: UBC Healthy Bones Studies. PMID- 15758319 TI - The structural adaptations of cortical bone to loading during different stages of maturation. PMID- 15758320 TI - The skeleton in a long-term perspective--are exercise induced benefits eroded by time? PMID- 15758321 TI - What is the prescription for healthy bones? PMID- 15758323 TI - Overview of estrogen action in osteoblasts: role of the ligand, the receptor, and the co-regulators. PMID- 15758324 TI - Direct and indirect effects of estrogen on osteoclasts. PMID- 15758326 TI - The role of estrogen metabolism in aging. PMID- 15758325 TI - ER-independent actions of estrogen and estrogen metabolites in bone cells. PMID- 15758327 TI - Novel paradigm on the effect of estrogen on bone. AB - According to prevailing unitary model of involutional osteoporosis, female postmenopausal bone loss can be divided into two separate phases: the accelerated, transient phase, which is most distinct over the subsequent decade after the menopause and accounts for 20-30% of the cancellous bone loss and 5-10% of the cortical bone loss (type I osteoporosis), and the following gradual, continuous bone loss (type II osteoporosis). Estrogen deficiency is currently quite unanimously accepted as the primary cause of type I osteoporosis, as well as also a major determinant of type II osteoporosis, and quite plausibly, the quest to uncover the origin of type I (and II) osteoporosis has focused on the estrogen withdrawal-related skeletal changes at and around the menopause. However, given that the cyclical secretion of estrogen begins normally in early adolescence and continues over the entire fertile period (excluding the potential periods of pregnancy) until the eventual cessation of female reproductive capability, one could argue that this menopause-oriented approach is limited in scope. In this review, some classic findings of the pubertal effects of estrogen on female bones are presented, findings that were paramount to Fuller Albright when he first described the disease called postmenopausal osteoporosis in 1940, but studies/findings that have failed to attract the attention they deserve. When these findings are incorporated with the primary function of the axial skeleton and long bones, the locomotion, an alternative, novel explanation for the function of estrogen and accordingly, the origin of the accelerated phase of postmenopausal bone loss, is proposed: estrogen packs mechanically excess bone/mineral into the female skeleton at puberty, a bone stock that later serves as the origin of the type I postmenopausal osteoporosis. PMID- 15758329 TI - Pathophysiology and epidemiology of TMJ. PMID- 15758331 TI - Biology and pathology of the temporomandibular joint: is there an animal model to study this affliction? PMID- 15758330 TI - TMJ anatomy and animal models. PMID- 15758332 TI - Does the relaxin, estrogen and matrix metalloproteinase axis contribute to degradation of TMJ fibrocartilage? PMID- 15758334 TI - Introduction--Bone turnover and fracture risk. PMID- 15758335 TI - Periosteal apposition and fracture risk. PMID- 15758336 TI - Transmenopausal and age-related changes in bone remodeling. PMID- 15758337 TI - Clusters and composites: Bone turnover and femoral neck fragility. PMID- 15758340 TI - Relevance of bone mineral density, bone quality and falls in reduction of vertebral and non-vertebral fractures. AB - All epidemiological studies conclude that without prompt, concerted and well designed prevention programs, the increasing cost related to osteoporotic fractures will become an unbearable burden for the community within the next fifteen years. However, the most effective way of setting up such preventive strategies is not yet unequivocally defined. Low bone mass and microarchitectural damage of bone tissue may account for a large part of the epidemiology of vertebral fractures. Extraskeletal determinants, including low muscle strength, poor balance and gait, all resulting in an increased propensity to fall, also play a major role in the occurrence of hip fracture. Depending on the localization of the fractures, the relative importance of skeletal and extraskeletal risk factors can significantly differ. For prevention of vertebral fractures, drugs affecting bone mass and skeletal architecture may provide a substantial benefit while hip fracture prevention will be more successfully targeted by multi-faceted strategies concentrating not only on the skeletal dimension of the fracture but also aiming, either pharmacologically or through multi-intervention programs, at a reduction in the incidence and in the consequences of falls in the elderly. PMID- 15758341 TI - Calcium homeostasis and bone surface proteins, a postulated vital process for plasma calcium control. AB - This report is a more in-depth explanation of a recently reported hypothesis for controlling the ionic calcium content of plasma and extracellular fluids (ECF). The hypothesis proposes a two-step process for returning calcium to the ECF against the established gradient continuously moving calcium from plasma to bone surfaces. The first step in this process is the predicted transfer of calcium directly from bone surfaces to the non-collagenous proteins, which are in contact with bone mineral. This calcium would be complexed to existing proteins and a portion would automatically become available for equilibration with ionic calcium in the ECF. The basis of the hypothesis is that the equilibration level helps to set the ionic calcium concentration of plasma. The gradient toward bone and the proposed two-step return occur in the ECF of bone and would be considered normal physiochemical processes. Thus, these processes are critical for mineral ion homeostasis in mammals. In this hypothesis, parathyroid hormone (PTH) is not required for the basic process. However, PTH works within the process to raise and set a precise plasma calcium concentration. The report to follow describes the process and discusses its relationship to normal and pathological conditions affecting human health. PMID- 15758342 TI - New targets for fascial, ligament and tendon research: a perspective from the Utah paradigm of skeletal physiology. AB - Here an octogenarian voice from the past argues that the physiology of fascia, ligaments and tendons has strong and useful analogs in some general features of bone physiology, including the latter's tissue-level mechanostat. Such analogs could provide unusually important targets for future collagenous-tissue research. Even by 2002, this field's authorities seldom discussed those analogs so this text concerns them. How well the above research proceeds could depend partly on A) making collagenous- tissue physiologists aware of those analogs (hence this article), B) on following a four-step analytical strategy, C) on "connecting the dots" between evidence and ideas from many clinical and basic-science fields to find larger "messages" and patterns hidden in mountains of lesser things, D) and on combining (i), cell- and molecular-biologic work, expertise and insights, (ii) with live-animal research and expertise and insights of the Utah paradigm of skeletal physiology. Why the "in vitro/in vivo collaboration" in "D" above? Partly because few, if any, skeletal tissue-level mechanisms function normally in current cell, tissue and organ culture systems. Consequently and historically an agent's in vitro effects seldom predicted correctly its in vivo effects, although the former effects may help to explain the latter ones after other studies revealed the latter ones. Things summarized in this article provide a foundation on which to build in the future. Since aging and other things took me out of that "building game", younger people will do that building when and how, and if, they wish to. The directions for that building suggested in this article differ enough from currently accepted "wisdom" that it may take years for most physiologists to concede their merit and begin that building in earnest. If so, so be it. PMID- 15758343 TI - Osteoporosis management in day-to-day practice. The role of calcitonin. PMID- 15758344 TI - Alterations in skeletal and mineral metabolism following thermal injuries. AB - Severe burns and other chronic inflammatory diseases are associated with altered skeletal metabolism that result in an increased incidence of osteopenia. In thermally injured children and adults there is a dramatic decrease in bone formation accompanied with an increase or maintenance of bone resorption. Children also exhibit a growth delay and subsequently fail to reach a predicted stature. Animal models, including the thermal injury mouse model, are being used to understand the mechanisms behind the uncoupling of bone formation and resorption that occurs following a major burn. The model has numerous commonalities with the human condition such as reduced bone formation, increased bone resorption, and decreased endochondral growth. The mechanisms that modulate calcium and skeletal metabolism following a thermal injury are complex and likely involve a number of endocrine, cytokine, and immune factors. Specifically, the potential roles of glucocorticoids, growth hormone, insulin-like growth factor-1, parathyroid hormone, interleukin-1 and -6, and tumor necrosis factor alpha are addressed. Subsequent to the increased survival rate of burn victims, there has been a heightened focus on therapeutic interventions that prevent or decrease the impact of thermal injuries on the skeletal system. These include exercise programs, exogenous recombinant human growth hormone, insulin, and oxandrolone. PMID- 15758345 TI - Effects of physical training on proprioception in older women. AB - Older adulthood is accompanied by declines in muscular strength, coordination, function, and increased risk of falling. Resistance training increases muscular strength in this population but its effect on proprioception is unknown. To evaluate the effect of resistance training on proprioception, community dwelling older women completed a three-month exercise study. A resistance training (RT) group (N=19) underwent supervised weight training three times per week while a non-strength trained control (NSTC) group (N=19) performed range-of-motion activities that mimicked the movements of the RT group without the benefit of muscle loading. Subjects were evaluated at baseline, 6, and 12 weeks for strength and proprioception. Muscular strength was assessed by measuring the subject's one repetition maximum performance on four different exercises. Static proprioception was measured by the subject's ability to reproduce a target knee joint angle while dynamic proprioception was measured by the subject's ability to detect passive knee motion. The RT group made significant strength improvements compared to the NSTC group. Proprioception significantly improved in both groups by 6 weeks. Our findings suggest that improvements in proprioception can be obtained via regular activity that is independent of heavy muscle loading. PMID- 15758346 TI - Age-associated changes in quantitative ultrasonometry (QUS) of the os calcis in Lebanese women-assessment of a Lebanese reference population. AB - OBJECTIVE: This study was aimed to assess age changes in quantitative ultrasonometry (QUS) in a large sample of Lebanese women to determine a Lebanese reference population. DESIGN: Cross-sectional study. SUBJECTS AND METHODS: Broadband ultrasound attenuation (BUA) and speed of sound (SOS) and the stiffness index (SI) of the os calcaneus was measured in 4,320 women with a mean age of 52.5 years (age range 20 to 79 years) using three identical Achilles Express (GE/Lunar) and one Achilles Plus (GE/Lunar) ultrasonometry devices. Women were randomly selected and asked to participate in a nationwide screening program using the media, conferences, telephone calls etc. Measurements were performed at Red Cross centers located all over the country. No inclusion or exclusion criteria were used. RESULTS: There was an overall decline of 19.2% for BUA, 3.1% for SOS and 30.3% for SI between late adolescence and old age. In premenopausal women, BUA decreased only slightly by 3%, while postmenopausal women showed a significant decline of 16.2%. In contrast, SOS continuously decreased from the age of 42; there was a decline of 0.8% from adolescence to the menopause; postmenopausal women showed a larger decline of 2.4%. The SI of premenopausal women decreased by 6%, while postmenopausal women showed a significantly larger decline of 24.3%. SI value for the female Lebanese young adult reference is 8% lower than that of the American and European women (92 SI units compared to 100). At the age of 42, SI value for the Lebanese women is 10.4% lower than the American women and 7.5% lower than the European women (86 SI units compared to 96 and 93, respectively). At the age of 75, SI values for the Lebanese women is 4.4% lower than the American women and the European women (65 SI units compared to 68). The decline in stiffness index for the Lebanese women between age 20 and 75 years is about 30.3% compared to 32% for the American or European reference curves. The rate of decrease for the Lebanese women was 0.2 SI units per year for the premenopausal period, and 0.7 SI units per year for the postmenopausal period. CONCLUSION: The age-related female, Lebanese reference curve was significantly different from the American and the European reference curves used by the manufacturer. Therefore, the use of our standardized reference data instead of the proposed US or European database reduces the risk of overestimating osteoporosis in the Lebanese population. The impact of our results on the prevalence of osteoporotic fracture in Lebanon has to be evaluated later on. PMID- 15758347 TI - pQCT provides better prediction of canine femur breaking load than does DXA. AB - Our study was designed to examine the validity of dual energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT) measurements as predictors of whole bone breaking strength in beagle femora. DXA was used to determine the bone mineral content, bone area, and 'areal' bone mineral density. PQCT was used to determine the cross-sectional moments of inertia, volumetric densities of the bone, and to calculate bone strength indices based on bone geometry and density. A three-point bending mechanical test was used to determine maximal load. Three variables from the pQCT data set explained 88% of the variance in maximal load, with the volumetric bone mineral density explaining 32% of the variance. The addition of the volumetric cortical density increased the adjusted r(2) to 0.601 (p=0.001) and the addition of an index created by multiplying volumetric cortical bone density by the maximum cross sectional moment of inertia made further significant (p<0.001) improvements to an adjusted r(2) of 0.877. In comparison, when only the DXA variables were considered in a multiple regression model, areal bone mineral density was the only variable entered and explained only 51% (p<0.001) of the variance in maximal load. These results suggest that pQCT can better predict maximal load in whole beagle femora since pQCT provides information on the bone's architecture in addition to its volumetric density. PMID- 15758348 TI - Negative impact of Crohn's disease on bone mineral mass. AB - Prolonged chronic inflammation and corticosteroid therapy increase the risk of osteoporosis in patients with Crohn's disease. It has been estimated that 30% of these patients, who take steroids for prolonged periods, will suffer a vertebral fracture. Patients with Crohn's disease are difficult to wean from corticosteroids and therefore are at risk of developing bone complications. The purpose of this cross-sectional study was to examine the relationship between cumulative steroid dose, duration of the disease and the development of osteopenia in patients with Crohn's disease. We studied 28 patients (17 men, 11 women) with Crohn's disease: eight had one or more bowel resections and all the women were premenopausal. Serum calcium, phosphate, total alkaline phosphatase, immunoreactive parathyroid hormone (iPTH), 25(OH)Vitamin D(3) and 1,25 (OH)(2) Vitamin D(3) were measured by autoanalyser methods or radioimmunoassay. Bone mineral density (BMD) was studied using dual energy X-ray bone absorptiometry of the lumbar spine (L2-L4) and the femoral neck. Of these 28 patients, 27 received an average of 17.3 +/- 21.7 g (range 1 to 80) g of prednisone over a period of 4 to 216 months. Fourteen out of the 28 patients had mildly diminished bone density (z-score >-2.5 SD and < -1 SD) of the spine and 15/28 of the hip. We found a greater decrease in bone density (z-score < -2.5 SD) in 2 out of 28 patients at the spine and in 5 out of 28 at the femoral neck. Those in whom the duration of the disease was less than two years (12 patients) had significantly higher vertebral z-scores (-0.096 +/-0.91) than those who had the disease for over two years (-1.31 +/- 2.37), (p<0.05). We found no significant correlation between lumbar spine and femoral neck z-scores and cumulative steroid therapy. Six out of 28 patients (four women and two men), of mean age 47.2+/-11.7, had one vertebral fracture. The mean cumulative dose of steroids (prednisone or budesonide) in patients with vertebral fractures was higher but not significantly different from that in patients without fractures -20.1+/-18.2 versus 14.1+/-11.2 g of prednisone, respectively (p>0.05). No correlation was found between various serum hormones and other biochemical parameters of bone turnover or bone density. We conclude that a large proportion of patients with Crohn's disease have reduced bone mineral density (58% at the spine and 75% at the femoral neck). The pathogenesis of bone loss is probably multifactorial. Although steroid therapy might be an important contributory factor, we were unable to find a significant correlation between it and bone loss. On the contrary, we observed that the duration of the disease makes a significant contribution to bone loss. PMID- 15758350 TI - What is new in neuro-musculoskeletal interactions? PMID- 15758349 TI - The pathogenetic influence of I-parathyroid hormone on slipped capital femoral epiphysis. Towards a new etiologic approach? AB - The displacement of the femoral head along the upper femoral physis that occurs during adolescence or slipped capital femoral epiphysis (SCFE) is not a very common traumatic entity. Ever since Muller1 first described it in 1888, its symptoms, clinical manifestations, diagnosis, treatment and complications have been thoroughly described and studied. Nevertheless little progress has been accomplished as far as its etiology is concerned. In order to assess the potential pathologic influence of any parathyroid hormone (PTH) disturbances on the development of SCFE, we conducted a prospective clinical study with 14 patients, 7 boys and 7 girls (16 hips), suffering from SCFE (Group A). Another 5 patients who had been treated for SCFE a few years before the study, were used as a control group (Group B). We measured the level of I-PTH along with serum calcium (Ca) and phosphorus (P) levels. Furthermore, we checked all the necessary anthropometric characteristics of the patients (i.e., age, height, weight and sexual maturation). Each patient of Group A was categorized from grade I to grade V according to the progress of the slipping. The results showed an increased incidence of serum PTH level abnormalities (both decrease and increase) in Group A while Group B patients had normal results. The detected I-PTH serum level abnormalities were not in any pattern related to the Ca and P serum levels. We believe that a temporary parathyroid hormone disorder during the early years of adolescence may play a potentially significant role (along with other etiologic factors) in the development of SCFE. PMID- 15758351 TI - Skeletal implications of prostate cancer. AB - This manuscript reviews the theories behind the propensity of prostate cancer to cause bone metastases and skeletal implications of the prostate cancer biology and treatment modalities. The escape of tumor cells from the primary tumor in the prostate to secondary tumor sites in the axial skeleton probably occurs before the primary tumor is detected. Several theories offer explanations for the observed proclivity of prostate tumors to selectively colonize the axial skeleton. The interaction between the tumor cells and cells that populate bone marrow, in particular osteoblasts and osteoclasts, is important for creating a 'fertile' environment where tumor cells can establish and grow. Prostate cancer cells are capable of producing growth factors that can affect both osteoblasts, resulting in osteoblastic bone formation, and osteoclasts, resulting in excessive bone resorption. In addition to the capability to progress from testosterone dependent to testosterone-independent phenotype, the hallmark of metastatic prostate cancer is osteosclerosis similar to one induced experimentally in nude rats using CWR22 human prostate cancer cell line. Metastatic bone disease caused by excessive bone formation and bone resorption is the major cause of morbidity in patients with prostate cancer. The most common symptoms include pain, pathological fractures, spinal cord compression, cranial nerve palsies, bone marrow suppression and hypercalcemia. The introduction of prostate-specific antigen in clinical practice created a shift to where more prostate cancer patients with early disease receive androgen ablation treatment, which in return causes more bone loss and cancer-associated osteoporosis. Introduction of third generation bisphosphonates to treat skeletal consequences of malignancy further stressed the important interaction between the bone marrow stroma and cancer cells. Nevertheless, animal models and human prostate tumor cell lines that mimic all aspects of skeletal conditions in prostate cancer patients including osteoblastic bone response are needed to develop and screen for novel therapeutic and diagnostic modalities. PMID- 15758352 TI - Perspective: genetic and hormonal roles in bone disorders: insights of an updated bone physiology. AB - In 1997 Professor J. Gorski suggested endocrinology needed new paradigms (Endocrine News 1997; 22:4,12). 'Connecting the dots' between diverse facts and ideas drawn from many lines of inquiry, plus accumulating evidence and increasing inadequacies of earlier ideas and terminology, led to an updated bone physiology called the 'Utah paradigm' that reveals new genetic and hormonal potential roles in bone physiology and disorders. One way to find a bone disorder's cause(s) and treatment( s) could depend on understanding the underlying physiology well enough to design effective drugs for it. In early views cell-level effects on osteoblasts and osteoclasts could explain most endocrine and genetic roles in bone disorders. The updated bone physiology supplements those views with roles of bone's tissue-level 'nephron-equivalent' mechanisms (NEMs) and their functions (NEFs), including some roles of biomechanics, whole-bone strength and muscle strength. That updated physiology reveals at least 42 nexuses above the cell level, some of them extraosseous, where genetic and/or hormonal effects might cause or help to treat varied bone problems. That multifactorial physiology also suggests that in vivo skeletal phenomena usually depend on many interlocking, laddered and nested feedback systems. Due to lack of study, how genes and hormones affect those nexuses and feedback systems still remains nearly unknown. Because studies of bone physiology in in vitro systems seldom if ever correctly predicted the in vivo effects, further live-animal research should seek the in vivo effects. This article suggests why more of that kind of research is needed, and some directions it could take. PMID- 15758353 TI - On the strength-safety factor (SSF) for load-bearing skeletal organs. AB - The strength of healthy postnatal mammalian load-bearing bones, growth plates, joints, fascia, ligaments and tendons exceeds the minimum strength needed to keep voluntary mechanical usage from breaking or rupturing them or from causing arthroses. Thus, they have a strength-safety factor (SSF). Some general features of the physiology in the Utah paradigm of skeletal physiology can explain two things: (i) Why load-bearing bones should have an SSF, (ii) and why its numerical value should approximately 6 in healthy young adult mammals. The number and kinds of studies and facts that revealed those two things for load-bearing bones do not yet exist for the extraosseous load-bearing organs that are made with cartilage and collagenous tissue. However, clinical-pathologic observations suggest the latter organs' SSFs should depend on features analogous to those that create SSFs for load-bearing bones. If so, the physiology on which bone's SSF depends could suggest directions for future studies of the SSF determinants of load-bearing extraosseous organs. Biomechanicians currently favor strain above stress when discussing biomechanical roles in the functional adaptations of bones to mechanical loading. However, an SSF is best expressed in stress terms, so a Table in this article provides corresponding strain/stress/unit-load values for bone's three important thresholds, and for its ultimate strength. PMID- 15758354 TI - Osteoporosis: combination therapy for better or worse. AB - Osteoporosis is a condition that is associated with an increased susceptibility for fractures. In the past few years, several drugs have become available that can reduce the incidence of fractures in patients with osteoporosis. Since these drugs work through different cellular mechanisms, combining agents of different classes may have an additive or multiplicative effect on fracture risk reduction. Combination treatments that have been evaluated in clinical trials include bisphosphonates with estrogen, raloxifene or PTH/ bisphosphonates and PTH/ estrogen. In general, these trials have shown increases in bone mineral density over that observed with each agent alone. However, whether anti-fracture efficacy is improved, or worsened remains to be established. This article reviews the combination treatments that have been evaluated in clinical trials, with a discussion of the potential benefits and risks that those treatments entail. Integrating safety and cost issues will eventually determine whether those combinations will become the standard of care. PMID- 15758355 TI - Intra-tibial injection of human prostate cancer cell line CWR22 elicits osteoblastic response in immunodeficient rats. AB - We investigated the utility of CWR22 human prostate cancer cells for modeling human metastatic prostate cancer, specifically their ability to induce bone formation following intra-tibial injections in the nude rat. Prostate cancer is unique in regard to its tropism for bone and ability to induce new bone formation. In contrast to humans, other mammalian species rarely develop prostatic cancer spontaneously upon aging and do not have the propensity for bone metastasis that is the hallmark of cancer malignancy in men. We chose human prostate cancer cell line CWR22 based on its properties, which closely resemble all of the features that characterize the early stages of prostatic cancer in human patients including slow growth rate, hormone dependence/independence and secretion of prostate-specific antigen. When CWR22 cells were injected directly into the proximal tibia of immunodeficient male rats, both osteoblastic and osteolytic features became evident after 4 to 6 weeks, with elevated levels of serum prostate-specific antigen. However, osteosclerosis dominates the skeletal response to tumor burden. Radiological and histological evidence revealed osteosclerotic lesions with trabeculae of newly formed bone lined by active osteoblasts and surrounded by tumor cells. Toward the end of the 7-week study, osteolytic bone lesions become more evident on X-rays. Paraffin and immunohistochemical evaluations revealed mature bone matrix resorption as evidenced by the presence of many tartrate resistant acid phosphatase positive multinucleated osteoclasts. We conclude that the CWR22 human prostate cell line used in an intra-tibial nude rat model provides a useful system to study mechanisms involved in osteoblastic and osteolytic bony metastases. This type of in vivo model that closely mimics all major features of metastatic disease in humans may provide a critical tool for drug development efforts focused on developing integrated systemic therapy targeting the tumor in its specific primary or/and metastatic microenvironments. In addition to targeting bone marrow stroma, this strategy will help to overcome classical drug resistance seen at the sites of prostate cancer metastasis to bones. PMID- 15758356 TI - A potential role for gap junctions in breast cancer metastasis to bone. PMID- 15758357 TI - Effects of aging and exercise on density and cross-sectional area of femur in senescence-accelerated mouse prone 6. AB - The densities and cross-sectional areas (CSAs) of the femur in 40- and 60-week old senescence-accelerated mice prone 6 (SAMP6) were determined using peripheral quantitative computed tomography (pQCT) and compared with those in age-matched control mice (ICR). In addition, the effects of 20 weeks of voluntary running exercise with increasing loads on femur density and CSA were examined in SAMP6. There were no differences in the cortical and trabecular densities or CSAs between the 40- and 60-week-old ICR. The cortical and trabecular densities of the 60-week-old control SAMP6 were lower than those of the 40-week-old control SAMP6. There was no difference in the cortical CSA between the 40- and 60-week-old control SAMP6, while the trabecular CSA of the 60-week-old control SAMP6 was higher than that of the 40-week-old control SAMP6. It was concluded that an age related decline in femur cortical and trabecular densities occurs at an earlier age in control SAMP6, indicating that SAMP6 show accelerated progression of age associated osteoporosis. There were no differences in the cortical density between the 40-week-old control and 60-week-old exercised SAMP6. These data indicate that running exercise with increasing loads prevents age-associated osteoporosis in this senescence-accelerated mouse strain. PMID- 15758358 TI - Quinolone levels in serum and maxillofacial tissues under ibuprofen co administration following surgical trauma. AB - Administration of antibiotics is considered to be an important factor, during or after operational procedures in the maxillofacial area, in order to avoid post surgical complications. Furthermore, administration of anti-inflammatory drugs is often prescribed for control of the post-operative pain. The aim of this study was to determine the levels of quinolones in serum and tissues (parotid gland, tongue, mandible), during traumatic injury in the oral cavity, with or without co administration of ibuprofen, a non-steroidal anti-inflammatory drug. Four groups of Wistar rats, (A, B control), (C, D experimental) were used. In the experimental group, traumatic injury was performed through the whole length of the cheek. Groups B and D received ibuprofen. The quinolone levels in serum and tissues were estimated by the inhibition zone of B. subtilis. Free fatty acid (FFA) levels and the adrenal weight, considered as a stress index, were increased in trauma groups. Quinolone concentrations in serum and in most of the tissues were significantly higher in the experimental groups compared to the controls. However, the co-administration of ibuprofen caused a higher increase of the quinolone levels in the control animals than in the experimental groups. PMID- 15758359 TI - A micro-computed tomography study of the trabecular bone structure in the femoral head. AB - The goal of this study was to characterize the trabecular microarchitecture of the femoral head using micro-computed tomography (ICT). Femoral head specimens were obtained from subjects following total hip replacement. Cylindrical cores from the specimens were scanned to obtain 3-D images with an isotropic resolution of 26 Im. Bone structural parameters were evaluated on a per millimeter basis: relative bone volume (BV/TV), trabecular number (Tb.N), thickness (Tb.Th) and separation (Tb.Sp), structure model index (SMI), and connectivity (Conn.D). The ICT data show that the first two millimeters, starting at the joint surface, are characterized by more plate-like trabeculae, and are significantly denser than the underlying trabecular bone. Regional differences in the trabecular architecture reveal that the superior pole has significantly higher BV/TV, Tb.N and Tb.Th values, with lower Tb.Sp compared to the inferior and side poles. Because subchondral bone is essential in the load attenuation of joints, the difference in bone structure between the subchondral and trabecular bone might arise from the different functions each have within joint-forming bones. The denser trabecular structure of the superior pole as compared to the inferior pole can be interpreted as a functional adaptation to higher loading in this area. PMID- 15758360 TI - On the pathogenesis of osteogenesis imperfecta: some insights of the Utah paradigm of skeletal physiology. AB - The pathogenesis of osteogenesis imperfecta (OI) baffled physiologists and physicians for over a century. Most past efforts to explain it depended heavily on cell and molecular biology and on changes in the material properties of affected bones (an old idea that OI patients could not make enough bone erred). To such views the still-evolving Utah paradigm of skeletal physiology can add a model for bone and bones that depends on errors in three genetically-determined features. The errors include, 1,2) elevated 'set points' of the strain-dependent thresholds that help to control how lamellar bone modeling and remodeling adapt bone strength, architecture and 'mass' to the voluntary loads on load-bearing bones; 3) and a reduced modeling-rate limit for the appositional rate of the lamellar bone formation drifts that can increase bone strength, outside bone diameters, cortical and trabecular thickness, and bone 'mass'. If only abnormalities #1,#2 occurred, that should limit the eventual strength, architecture and 'mass' of load-bearing bones, while if only #3 occurred that should prolong or delay how long it took to achieve the above limits, but without changing them. Equally, in driving from New York to Boston, stopping at New Haven would prevent reaching Boston no matter how rapidly one drove (a limited trip). But by not stopping one could reach Boston by driving very slowly (a prolonged but not a limited trip). This model concerns general features of bone and bones in OI that would need study and explanation at the tissue, cellular and molecular biologic levels. Other places and people must discuss any devils in the details, as well as collagenous tissue, auditory, dental and other problems in OI, and the effects of treatment on the above features. PMID- 15758361 TI - Myostatin expression in age and denervation-induced skeletal muscle atrophy. AB - Myostatin is hypothesized to regulate skeletal muscle mass and to be a potential target for therapeutic intervention in sarcopenia. To clarify whether myostatin is invariably associated with sarcopenia, this study examined the levels of expression of myostatin mRNA and protein in Sprague Dawley rats during aging- and denervation-induced sarcopenia. The level of myostatin mRNA in the gastrocnemius decreased progressively with age being 9, 34 and 56% lower at 6, 12 and 27 months, respectively, compared with mRNA levels at 1.5 months. In contrast, two low molecular mass isoforms of myostatin protein identified by Western blotting increased progressively with age. With denervation, myostatin mRNA was 31% higher on day 1 but by 14 days after sciatic neurectomy when the muscle had atrophied 50%, myostatin expression decreased 34% relative to the sham operated limb. Western analysis of the denervated gastrocnemius showed that myostatin protein levels varied in parallel with mRNA. These disparate patterns of expression of myostatin during age- and denervation-induced atrophy suggest that the regulation of myostatin is complex and variable depending on whether the atrophy is slowly or rapidly progressive. PMID- 15758362 TI - 'Pre-synaptic' vesicular glutamate release mechanisms in osteoblasts. AB - Identification of intercellular signalling pathways in bone represents an important therapeutic target for drug development in the treatment of clinical conditions such as osteoporosis. One such intercellular signalling pathway in bone appears to be mediated by the excitatory amino acid glutamate, exhibiting remarkable similarities to synaptic neurotransmission. Bone cells (osteoblasts and osteoclasts) express functional glutamate receptors that are electrophysiologically and pharmacologically similar to those expressed in the CNS and there is evidence for their involvement in both bone formation and bone resorption. However, to date the cellular source of glutamate for the activation of these specific glutamatergic receptors in bone has remained unclear. This review provides a synopsis of our current understanding of these 'pre-synaptic' signalling mechanisms, presenting compelling evidence that osteoblasts possess the molecular capability to direct regulated vesicular glutamate release in response to osteotropic regulatory inputs. In addition, we discuss mechanisms other than 'pre-synaptic' glutamatergic mechanisms that could account for the source of glutamate for receptor activation in osteoblasts. Finally, convincing evidence reporting physiologically released glutamate in varied osteoblasts and osteoblastic cell lines is discussed. The overwhelming conclusion of this review is that by defining both the characteristics and regulatory control of this process, highlighting both similarities and differences between the CNS and bone may provide compelling evidence for the role of glutamate in bone cell function and physiology. PMID- 15758363 TI - Breast cancer metastasis to bone: evolving models and research challenges. PMID- 15758364 TI - Alfacalcidol restores cancellous bone in ovariectomized rats. AB - Active vitamin D metabolites have been demonstrated to reduce vertebral and hip fractures in elderly patients. A number of in vitro and in vivo pre-clinical studies have suggested that vitamin D may effectively stimulate osteoblastic activity and exert an anabolic effect on bone. The current study was designed to further explore the ability of an active vitamin D analog to restore bone in a skeletal site with established osteopenia in ovariectomized (OVX) rats. Female Sprague Dawley rats at five months of age and 8 weeks after sham ovariectomy or ovariectomy were randomly divided into 7 groups with 10 per group. At the beginning of the treatments, one group of sham-operated rats and one group of OVX rats were sacrificed to serve as baseline controls. Another group of sham operated rats and one group of OVX rats were treated with vehicle for 4 weeks. The OVX rats in the remaining groups were treated with alfacalcidol at 0.05, 0.1 or 0.2 microg/kg/d by daily oral gavage, 5 days/week for 4 weeks. As expected, estrogen depletion caused high bone turnover and cancellous bone loss in lumbar vertebra of OVX rats. Alfacalcidol treatment at 0.1 or 0.2 but not 0.05 microg/kg/d increased serum calcium and phosphorus in OVX rats as compared with vehicle treatment. In addition, serum parathyroid hormone was suppressed, whereas serum osteocalcin was increased by alfacalcidol at all dose levels. Furthermore, histomorphometric data of 2nd lumbar vertebral body revealed that cancellous bone volume in OVX rats treated with alfacalcidol at 0.1 or 0.2 microg/kg/d was increased to the level of sham-operated rats treated with vehicle. This increment in cancellous bone mass was accompanied by increases in trabecular number and thickness and a decrease in trabecular separation. Moreover, osteoclast surface and number were significantly decreased, whereas bone formation variables such as mineralizing surface and bone formation rate were significantly increased in alfacalcidol- treated OVX rats compared with those of vehicle-treated OVX rats. Finally, a linear regression analysis showed that alfacalcidol treatment dose dependently altered most of the variables measured in the current study. In conclusion, alfacalcidol completely restores cancellous bone by stimulating bone formation and suppressing bone resorption in lumbar vertebra of OVX rats when the treatment is started at an early phase of osteopenia. The evidence of increased bone formation by alfacalcidol treatments further supports the notion that active vitamin D metabolites or their analogs may exert anabolic effects on bone. PMID- 15758365 TI - Progressive isometric force training and bone mass in rats. AB - In ten growing male Wistar rats, isometric strength training for 69 days (3-6 times vertical gripping position on a wire-netting during 2 x 30 s, with progressive loading of the tail through a 50-200 g indwelling clip), fat mass and plasma leptin concentrations were lower on day 70 than in ten sedentary controls. Muscle mass and femoral trabecular and cortical bone mineral density were simultaneously higher in exercised animals than in controls. Such an effect might result from decreased bone resorption. At the end of the training period no difference concerning plasma osteocalcin concentration was observed between exercised and resting rats while urinary deoxypyridinoline excretion was lower in the former than in the latter. PMID- 15758366 TI - Regulation of osteoclast differentiation by statins. AB - HMG-CoA reductase inhibitor (statin) treatment is frontline therapy for lowering plasma cholesterol levels in patients with hyperlipidemia. In a few case studies, analysis of clinical data has revealed a decreased risk of fracture in patients on statin therapy. However, this reduction in the incidence of fracture is not always observed nor is it supported by an increase in bone density, which further complicates our understanding of the role of statins in bone metabolism. Thus, the precise role of statins in bone metabolism remains poorly understood. In this study, we examined the effect of statin treatment on osteoclastogenesis. Treatment with lovastatin resulted in a significant, dose-dependent decrease in the numbers of differentiated osteoclasts and decreased cholesterol biosynthesis activity with an EC(50) similar to that observed in freshly isolated rat or cultured human liver cells. Studies assessing the role of mevalonate metabolites in the development of the osteoclasts demonstrated that geranylgeraniol, but not squalene or farnesol was important for the development and differentiation of osteoclasts, implicating protein geranylgeranylation rather than protein farnesylation as a key factor in the osteoclast differentiation process. In conclusion, our data indicate that lovastatin inhibits osteoclast development through inhibition of geranylgeranylation of key prenylated proteins and that the bone effects of statins are at least partially due to their effects on osteoclast numbers. PMID- 15758367 TI - Bone mass and density response to a 12-month trial of calcium and vitamin D supplement in preadolescent girls. AB - BACKGROUND: Maximal bone acquisition in adolescent girls through dietary and lifestyle practices is advocated to prevent or minimize the development of osteoporosis and its associated complications in later life. Longitudinal investigations of bone acquisition in children and adolescents have utilized areal bone mineral density (BMD, mg/cm(2)) as a measure of bone mass and strength. Peripheral quantitative computed tomography (pQCT), which provides a three-dimensional display of data, separate analyses of bone compartments, and bone mass in terms of volumetric BMD (vBMD, mg/cm(3)), has recently been introduced for clinical use. OBJECTIVE: To assess the impact of a 12-month daily calcium supplement on total and trabecular bone acquisition as measured by pQCT in preadolescent girls. DESIGN: Early adolescent Caucasian girls (aged 12 years, Tanner Stage 2) were enrolled in a randomized trial of daily calcium supplement (TX, 800 mg calcium carbonate and 400 IU vitamin D) or placebo (C). Body weight, height, and distal tibia measurements by pQCT were obtained at enrollment, 6 and 12 months. Pubertal status and physical activity records were assessed at baseline and 12 months. Three-day food intake records were completed every three months. RESULTS: Seventy-one girls completed the 12-month trial (TX=35, C=36). No differences were found for age, weight, height, body mass index, pubertal maturation, or reported physical activity at enrollment or during the study. Average intakes during the study were 1524 mg calcium and 496 IU vitamin D (TX) versus 865 mg calcium and 160 IU vitamin D (C) per day. Baseline total bone values were similar, however, trabecular values were greater in TX girls despite randomization. Percent changes were calculated to adjust for baseline differences. Because of the small cortical thickness at the 10% site (mean values < 1.6 mm), cortical mass and density were not analyzed. The percent changes for trabecular bone mineral content (BMC, mg) and vBMD were significantly greater in TX girls (+4.1% BMC and +1.0% vBMD TX versus -1.6% BMC and -2.0% vBMD C, p<0.006; ANCOVA) after 12 months of supplement. Trabecular bone area (BA, cm(2)) and total bone change, however, did not differ between groups. CONCLUSIONS: Daily calcium and vitamin D supplementation promotes greater trabecular BMC and vBMD acquisition in preadolescent girls. The single site selected for pQCT evaluation in this study did not allow evaluation of the cortical bone compartment. Future studies that utilize the pQCT technique need to incorporate multiple measurement sites to better assess total, cortical, and trabecular bone. PMID- 15758368 TI - Bone mineral density in hypoparathyroid women on LT4 suppressive therapy. Effect of calcium and 1,25(OH)2 vitamin D3 treatment. AB - Our aim was to study the bone mineral density (BMD) of patients with chronic hypoparathyroidism (hypoPTH) after longterm calcium and vitamin D treatment. Twenty hypoPTH women (mean-/+SD, aged 50-/+15 years, IPTH 4-/+6 pg/ml) and 20 matched euparathyroid women (euPTH) after near total thyroidectomy for thyroid cancer, completed with I-131 ablation and on suppressive therapy with L-Thyroxine (LT(4)), were studied. In addition eight hypoPTH patients who were receiving LT(4) replacement therapy after surgery for compressive goiter were simultaneously studied. The hypoPTH patients were on calcium and 1,25(OH)(2) vitamin D(3) therapy to normalize serum calcium. Bone mineral density (BMD) (DXA, at the lumbar spine [L(2)- L(4), LS], femoral neck [FN] and Ward triangle [WT]), serum and urine calcium, serum phosphorus, TOTALALP and osteocalcin were measured. Patients with hypoPTH showed greater lumbar BMD than euPTH patients on suppressive therapy (Z-score; 1.01-/+1.34 vs. -0.52-/+0.70, p<0.05). Serum osteocalcin levels were higher in hypoPTH patients on suppressive therapy compared to hypoPTH patients on replacement therapy. The LS BMD from hypoPTH patients correlated with calcium supplements (r=0.439; p=0.02), 1,25(OH)(2)D(3) dose (r=0.382; p=0.04) and LT(4) dose (r=0.374; p=0.05). Our data suggest that long-term treatment with calcium and 1,25(OH)(2) vitamin D3 supplements in hypoPTH patients on suppressive LT4 therapy results in increased BMD when compared with patients with normal PTH levels. PMID- 15758369 TI - Long term effects of a single dose of intravenous Ibandronate. AB - For defining the optimal regimen of a treatment of osteoporosis with an intravenous bisphosphonate one needs to know the duration of action of a single dose of the given drug. This allows us to establish the frequency by which a given dose has to be administered for obtaining a sufficient suppression of bone resorption over a longer period. In this study 1 and 2 mg Ibandronate were given as a single i.v. injection to young normal men and to healthy postmenopausal women on a free diet and with no treatment or supplements, and the markers of bone metabolism, as well as BMD, were followed for 6 months. Urinary C telopeptides decreased by 81-95% 1 week after the injection and remained significantly decreased for 2 weeks after 1 mg Ibandronate, and for 4 months after 2 mg. In men, PTH increased by 80% at one week and remained significantly increased for 2 weeks, after 1mg and 2 mg Ibandronate. Plasma osteocalcin decreased slowly over 2 months in all 3 groups by 22%. Alkaline phosphatase showed similar, but not significant changes. In conclusion, the inhibition of bone resorption induced by 1 mg Ibandronate i.v. does not exceed 1 month and does not allow 3 month intervals in the treatment of osteoporosis, while 2 mg cover 3 months at least. PMID- 15758370 TI - Measurement of gene expression following cryogenic mu-CT scanning of human iliac crest biopsies. AB - An important consideration in interpreting indices of gene expression in human bone is relating mRNA levels to functional endpoints such as bone architecture. In the present study, a method was developed for quantitative measurement of gene expression and bone morphology in the same specimen. Three-dimensional images of iliac crest bone biopsies from healthy premenopausal women were obtained using a novel high resolution cryogenic mu-CT scanner. RNA was isolated from the biopsies and mRNA levels were measured for genes related to bone metabolism. The gene expression profile and variability of expression within iliac crest biopsies of women was similar to human osteoblastic cell lines and rat long bones. mRNA for alkaline phosphatase, bone matrix proteins, and selected cytokines and cytokine receptors were consistently detected in biopsies. As previously shown in rat bone, there was a tight correlation between mRNA levels for type 1 collagen and osteonectin, a weaker correlation between type 1 collagen and osteocalcin and no correlation between bone matrix proteins and alkaline phosphatase. The relative abundance of the mRNA for the three most prevalent transforming growth factor beta (TGF-beta) isoforms in bone (TGF-beta(1)>> TGF-beta(3)> TGF-beta(2)) was the same as the known abundance of the corresponding TGF-beta peptides in bone matrix. The results demonstrate the feasibility of analyzing the three dimensional architecture of a bone biopsy using cryogenic mu-CT imaging and then measuring expression of genes related to bone cell function within the same specimen following RNA extraction and analysis. PMID- 15758371 TI - Enthesial fibrocartilage - bone interaction: a radiographic study of selected sites of nonsynovial peripheral enthesopathy. AB - OBJECTIVES: To assess radiological imaging and prevalence of pelvic (excluding sacroiliac joints), calcaneal, patellar and humeral enthesopathy (EN) in a cohort study. MATERIALS-METHODS: Outpatients attending a state hospital rheumatology clinic for a continuous 4-year period, were consecutively screened for radiographic evidence of peripheral nonsynovial EN in pelvis, ankle, knee and shoulder regions and in particular sites within these regions regardless of symptoms. Imaging and prevalence were assessed in patients suffering from a variety of myoskeletal disorders by applying either of the following two plain X ray criteria: a) tendon and/or ligament and/or fascia ossification, b) reactive bone proliferation resulting in excrescences and/or sclerosis and/or erosions. RESULTS: A total of 3,670 patients were screened and a cohort of 585 patients (16%) with extraspinal peripheral EN was selected. Plain radiography provided good imaging of pelvic EN at iliac crests, greater trochanters, pubic symphysis and pubic rami, as well as of calcaneal, patellar and humeral head enthesopathic changes. Cohort recruitment by applying the two aforementioned criteria resulted in the formation of 2 groups: Group A, consisting of 169 patients (mean age in years 34 -/+ 8 SD) suffering from inflammatory myoskeletal disease represented by Seronegative Spondyloarthropathies (SSp); and Group B, including 416 patients (mean age 63 -/+ 7) suffering from degenerative/metabolic disorders classified as degenerative disease of the spine, hip or knee (70%), Diffuse Idiopathic Skeletal Hyperostosis (DISH) (11%) and rotator cuff (Rot/Cuff) syndromes (19%). Females were the predominant gender in the cohort and in Group B patients (both p<0.001 vs. males), while the opposite was true for the group of inflammatory diseases. Patients in Group A were younger and had shorter disease duration than those of group B (p<0.001 for both). Pelvic EN was the most frequent localization of EN within the cohort (46%, p<0.001) followed by both multiple site and patellar EN (24% and 22% respectively). Patients in Group A, had a significantly higher prevalence of pelvic EN compared to those in Group B (60% vs. 39%, p<0.001) and the former group was significantly associated with pelvic EN. On the contrary, although pelvis was also the predominant EN site in Group B, patellar and humeral head EN were significantly associated with noninflammatory diseases. In patients with SSps, pelvic EN predominance (60%) was followed by calcaneal involvement (p<0.01 vs. patellar and humeral head). These two were the skeletal sites that were significantly associated with individual diseases within Group A (pelvis with AS and Ps-Sp and calcaneus with RR). Within Group B, patients with knee OA, hip OA and Rot/Cuff showed EN site localization in absolute proximity with disease process, while in those with Deg/Sp or DISH pelvis was the predominant site involved. CONCLUSIONS: Plain radiography provides good imaging of peripheral nonsynovial EN at well defined skeletal sites. Within a general rheumatic population, pelvic EN is the most prevalent localization followed by multiple site and patellar reactive bone lesions. Apart from seronegative spondyloarthopathies, degenerative and metabolic myoskeletal disorders contribute substantially to local induction of abnormal fibrous tissue/fibrocartilage-bone interactions resulting in radiographically detectable EN. PMID- 15758372 TI - What is new in neuro-musculoskeletal interactions? PMID- 15758373 TI - The 32nd Sun Valley Hard Tissue Workshop. Introduction. PMID- 15758374 TI - Life history of osteocytes: relationship to bone age, bone remodeling, and bone fragility. PMID- 15758375 TI - Animal models of osteoarthritis in an era of molecular biology. AB - Animal models of osteoarthritis (OA) are used to study the pathogenesis of cartilage degeneration and to evaluate potential anti-arthritic drugs for clinical use. In general, these models fall into 2 categories, spontaneous and induced (surgical instability or genetic manipulation). Animal models of naturally occurring OA occur in knee joints of guinea pigs, mice and Syrian hamsters. Commonly utilized surgical instability models include medial meniscal tear in guinea pigs and rats, medial or lateral partial meniscectomy in rabbits, medial partial or total meniscectomy or anterior cruciate transection in dogs. Transgenic models have been developed in mice. These models all have potential use in the study of molecular mechanisms associated with OA development via use of immunohistochemistry, biochemistry and molecular probes to identify altered matrix molecules at different stages in disease progression. Testing of specific types of inhibitors developed through evaluation of matrix changes in the disease process will ultimately help identify key processes which initiate and perpetuate the disease and will lead to discovery of new disease modifying pharmaceutical agents for OA patients. This paper will focus on the discussion of several models which are likely to be useful in the molecular dissection of processes involved in cartilage degeneration. PMID- 15758376 TI - Altered subchondral osteoblast cellular metabolism in osteoarthritis: cytokines, eicosanoids, and growth factors. PMID- 15758377 TI - Interactions between subchondral bone and cartilage in OA. Cells from osteoarthritic bone can alter cartilage metabolism. PMID- 15758380 TI - Molecular regulation of the chondrocyte phenotype. PMID- 15758379 TI - Perlecan: an important component of the cartilage pericellular matrix. AB - Perlecan (Pln) is a large proteoglycan that can bear HS (heparan sulfate) and chondroitin sulfate glycosaminoglycans. Previous studies have demonstrated that Pln can interact with growth factors and cell surfaces either via its constituent glycosaminoglycan chains or core protein. Herein, we summarize studies demonstrating spatially and temporally regulated expression of Pln mRNA and protein in developing and mature cartilage. Mutations either in the Pln gene or in genes involved in glycosaminoglycan assembly result in severe cartilage phenotypes seen in both human syndromes and mouse model systems. In vitro studies demonstrate that Pln can trigger chondrogenic differentiation of multipotential mouse CH310T1/2 stem cells as well as maintain the phenotype of adult human chondrocytes. Structural mapping indicates that these activities lie entirely within domain I, a region unique to Pln, and that they require glycosaminoglycans. We also discuss data indicating that Pln cooperates with the key chondrogenic growth factor, BMP-2, to promote expression of hypertrophic chondrocyte markers. Collectively, these studies indicate that Pln is an important component of human cartilage and may have useful applications in tissue engineering and cartilage-directed therapeutics. PMID- 15758383 TI - Osteoporosis and fracture risk: bone matrix quality. PMID- 15758381 TI - The vWFC domain of type IIA procollagen amino-propeptide functions as an antagonist of bone morphogenetic proteins. PMID- 15758384 TI - Determinants of skeletal fragility and bone quality. PMID- 15758385 TI - Changes in bone collagen with age and disease. PMID- 15758386 TI - Variations in bone mineral properties with age and disease. PMID- 15758387 TI - Architectural changes independent of bone mineral in osteoporosis. PMID- 15758388 TI - Effects of bisphosphonates on matrix mineralization. AB - Bone strength is determined not only by the volume of bone tissue and the microarchitectural organization of this bone, but also by the degree of mineralization of bone matrix. The mineralization process consists of a primary deposition of mineral substance on the calcification front, followed by a slow and progressive increase of the mineral deposition named secondary mineralization. In osteoporosis, there is a negative imbalance between bone resorption and bone formation, resulting in bone loss, and microarchitectural deterioration of the trabecular network. Therapeutic agents for osteoporosis could increase bone strength by three separate, but interrelated effects on bone tissue: 1) the prevention of bone loss and thus the preservation of bone microarchitecture, 2) an increase in the volume of bone matrix, and 3) an increase in the degree of mineralization to a level similar to that seen in healthy premenopausal women, through a prolongation of the duration of secondary mineralization. Therefore the use of antiresorptive agents that reduce bone turnover, as bisphosphonates, provide a rational approach to treatment of osteoporosis. Extensive phase III clinical trials have shown that osteoporotic women treated orally with alendronate (ALN) for 3 years or more had substantial increases in bone mineral density (BMD) of approximately 10% at the spine together with reductions of about 50% in the incidence of vertebral fractures. Since a marked reduction in activation frequency was evidenced in the transiliac biopsies taken after treatment with ALN compared to placebo (PLA), without detectable increase in cancellous bone volume, it was hypothesized that the increase in BMD and the reduction in the incidence of fragility fractures were due, in a substantial part, to an increase in the degree of mineralization of bone (DMB). The mean DMB was measured by quantitative microradiography on transiliac bone biopsies taken from 53 postmenopausal osteoporotic women who had been treated with ALN (10 mg/day) during 2 (9 patients) or 3 years (16 patients) or with PLA (15 and 13 patients, respectively). In the same patients, BMD values were obtained by dual-energy X-ray absorptiometry on lumbar spine at the beginning and end of treatment. Histomorphometric parameters and activation frequency of new remodeling units were also measured on the biopsies. After 2 years of ALN, mean DMB in compact bone was 9.3% (p=0.0035) and in cancellous bone was 7.3% (p=0.0009) higher, respectively, versus PLA. After 3 years of ALN, mean DMB in compact bone was 11.6% (p=0.0002) and in cancellous bone was 11.4% (p=0.0001) higher, respectively, versus PLA. After 2 and 3 years of ALN and compared to the corresponding PLA, the distribution of the DMB clearly showed a shift towards the highest mineralization values and a decrease of the number of bone structure units having low values of mineralization. The between group differences in mean DMB were similar to those of BMD at the lumbar spine level (+8.7% after 2 years +9.6% after 3 years, respectively), suggesting that mean DMB augmentation probably accounts for the major part of the increase in BMD seen with ALN. These results support our model that the reduction in the activation frequency caused by the antiresorptive effect of ALN is followed by a prolonged secondary mineralization which increases the percentage of bone structure units having reached a maximum degree of secondary mineralization and, through this mechanism, mean DMB. That these effects contribute to improved bone strength is demonstrated by the reduction in fracture incidence previously demonstrated in these patients. In conclusion, quantitative microradiography gives access to the mineral dimension of bone tissue which has been insufficiently taken into account until now as an important determinant of bone strength and quality of bone. PMID- 15758390 TI - Introduction: osteosarcoma session. PMID- 15758391 TI - Osteosarcoma: clinical practice and the expanding role of biology. PMID- 15758392 TI - Radionuclide-induced skeletal cancers. PMID- 15758393 TI - Genetic and molecular aspects of osteosarcoma. PMID- 15758394 TI - The involvement of prostaglandins in tumorigenesis, tumor-induced osteolysis and bone cancer pain. PMID- 15758396 TI - Cancer cell metastases. PMID- 15758397 TI - Overview of metastasis and metastases. PMID- 15758398 TI - The vicious cycle of bone metastases. PMID- 15758399 TI - Cancer of the prostate - implication for bone metastases. PMID- 15758400 TI - Applications of high-throughput methods to cancer metastases. PMID- 15758402 TI - Osteoblast progenitor fate and age-related bone loss. PMID- 15758403 TI - Aging and protein expression. PMID- 15758406 TI - In memory of Professor Lis Mosekilde. PMID- 15758405 TI - Neurotransmitters in bone. Introduction. PMID- 15758407 TI - In memory of Professor Aurelio Rapado. PMID- 15758408 TI - Glutamate transporters in bone. AB - In the central nervous system Na(+)-dependent glutamate transporters bind extracellular glutamate and transport it into cells surrounding the synapse, terminating excitatory signals. These glutamate transporters also function as ion channels. The glutamate transporter, GLAST-1, is expressed in the plasma membrane of osteoblasts and osteocytes and is the same molecular weight as in brain. Thus in bone cells GLAST-1 may transport glutamate or operate as a glutamate gated ion channel. A splice variant, GLAST-1a, is also expressed in bone. Hydropathy and Western blot analysis suggest GLAST-1a adopts a reversed orientation within the cell membrane. Sodium and potassium ion gradients drive glutamate transport but glycosylation, oxidation and phosphorylation modulate transporter activity. Reversal of GLAST-1a would alter these modifications varying its transport activity under the same ionic gradients. The significance of GLAST-1/1a in bone in vivo is unknown. GLAST-1 knockout mice show no major disruption of skeletal development but have not been investigated in detail. Glutamate affects both osteoclast and osteoblast biology and the regulation of GLAST-1 by mechanical loading in bone suggests a role for glutamate transporters in osteogenesis. Differential regulation and modification of GLAST variants may provide an intricate mechanism controlling extracellular glutamate levels and thus its downstream signalling effects in bone. PMID- 15758409 TI - Functional osteoblastic ionotropic glutamate receptors are a prerequisite for bone formation. AB - Many studies have now demonstrated that osteoblasts express the protein components necessary for functional neuronal-like glutamatergic signalling to occur, and as a result a physiological role for receptor mediated osteoblastic glutamate signalling has been proposed. Osteoblastic ionotropic type glutamate receptors (iGluRs) have been shown to be functional; they possess electrophysiological characteristics similar to neuronal iGluR) and agonist application modulates the activity of intracellular signalling molecules and osteoblastic transcription factors. The physiological importance of osteoblastic iGluRs is illustrated by the fact that osteoblasts treated in vitro with non competitive iGluR antagonists fail to form mineralized bone. Interestingly compounds known to antagonize specific sub-types of iGluR induce different effects when applied to osteoblasts derived from long and flat bones. These data imply that not only are functional osteoblastic iGluRs a prerequisite for osteoblast differentiation and bone formation, but also that the components of osteoblastic glutamatergic signalling may be adapted to reflect the differential function of osteoblasts from different skeletal sites. This paper reviews the evidence that suggests that iGluR-mediated glutamate signalling plays a fundamental role in the regulation of osteoblast function and bone formation, and discusses the therapeutic potential of manipulation of osteoblastic iGluR to modulate bone homeostasis. PMID- 15758410 TI - Glutamatergic regulation of bone resorption. AB - There has been increasing evidence during the last years that glutamate (Glu), the major neuromediator of the nervous system, contributes to the local regulation of bone cell functions. Several classes of Glu receptors and transporters, as well as molecules involved in glutamate signal transduction in neuronal tissue, were identified in bone. While recent findings suggest that Glu may participate in mechanisms underlying bone formation, several studies indicate that Glu may also control bone resorption. Ionotropic NMDA and metabotropic Glu receptors are expressed by osteoclasts and electrophysiological studies have demonstrated that NMDA receptors (NMDAR) are functional on these cells. In vitro studies have shown that NMDAR are important for osteoclast function since several specific antagonists of NMDAR which block the current induced by Glu in these cells also inhibit bone resorption. Preliminary studies investigating the mechanisms of action of NMDAR antagonists on bone resorption are reviewed in this paper. There is also growing evidence that NMDAR are expressed throughout the osteoclastic differentiation sequence and that antagonists of NMDAR affect osteoclastogenesis. Very few in vivo studies have however investigated the role of Glu in skeletal metabolism and bone resorption and clearly further work is required to demonstrate the relevance of glutamate signaling in the physiology of bone resorption in vivo. PMID- 15758411 TI - Expression and function of P2 receptors in bone. AB - ATP (adenosine 5'-triphosphate) is one of the most important extracellular regulatory molecules in the skeleton. Extracellular ATP and other nucleotides signal through P2 receptors, a diverse group of receptors that are widely expressed by bone cells. P2 receptors are divided into two subclasses; P2Y G protein coupled receptors, and P2X ligand-gated ion channels, and there is functional and molecular evidence for the expression of these receptors on both osteoblasts and osteoclasts. In order to activate P2 receptors, nucleotides must be released into the bone microenvironment. ATP is present in mmol concentrations in cells and can be released by cell lysis, cell trauma or physiological mechanisms, possibly through ABC transporters. Following co-activation of P2Y and PTH1 receptors on osteoblasts, there are multiple levels of interaction in downstream signalling that eventually lead to synergistic expression of osteoblastic genes, providing a mechanism for integrating local and systemic regulatory signals in bone particularly with regard to the activation of bone remodelling. Activation of P2Y1 receptors on osteoblasts enhances expression of RANKL leading indirectly to an increase in osteoclast formation and resorption. Expression of P2X7 inducible pores on osteoclast precursor cell membranes allows fusion to form multinucleated osteoclasts and blockade of this receptor inhibits resorption. The capacity of extracellular nucleotides to provide a highly localized and transient signal coupled with the profound effects of P2 receptor activation on osteoblastic and osteoclastic cells and the synergistic interactions with systemic hormones, indicate that nucleotides have a strong influence over bone tissue growth and regeneration. PMID- 15758412 TI - Neuropeptidergic regulation of bone resorption and bone formation. AB - Immunohistochemical studies have revealed an extensive network of nerve fibers in the vicinity and within the skeleton, not only in the periosteum of bone but also in cortical and trabecular bone as well as in the bone marrow. Phenotyping of the skeletal nerve fibers have demonstrated the expression of a restrictive panel of different signalling molecules including neuropeptides, neurotransmitters and neurotrophins. In this review, the presence of receptors for the neuropeptides vasoactive intestinal peptide, calcitonin gene-related peptide and substance P on osteoblasts and osteoclasts and the capacity of these receptors to regulate bone formation, osteoclast formation and activity are described. These findings, together with data obtained by chemically and surgically targeted nerve deletion and observations made in paraplegic patients, strongly suggest that neuro osteogenic interactions play an important role in skeletal function. PMID- 15758413 TI - Short-term rigid and flaccid paralyses diminish growth of embryonic chick limbs and abrogate joint cavity formation but differentially preserve pre-cavitated joints. AB - The influence of movement on joint space formation during limb development has been the subject of much interest. Our aim was to investigate the short-term influence of movement upon cavitation by immobilizing chick embryos in ovo, both in a rigid manner where dynamic stimulation is removed, and a flaccid manner where both dynamic and static stimulation are absent. Induction of rigid immobilization with decamethonium bromide (DMB) or the novel induction of flaccid immobilization with pancuronium bromide (PB) for 3 days, during the normal cavitation of joints resulted in the loss of cavity formation. Immobilization after the formation of an overt cavity demonstrated that static stimulation (during rigid paralysis) was able to maintain joint cavities and preserve some of the hyaluronan (HA) content of articular surfaces, whereas flaccid paralysis resulted in the loss of cavities and a marked depletion of HA content. Assessments of the growth and deposition of cartilage and bone in the limbs of embryos immobilised during cavitation showed that the length of limb elements was greatly reduced and that decreases in epiphyseal widths were most marked and more pronounced distally. The volume of bone in these elements remained unchanged whereas the cartilage volume decreased significantly, suggesting that chondrogenic but not osteogenic events in the embryo are particularly sensitive to mechanical stimulation. In addition to describing a novel method of inducing flaccid immobility in ovo, these data point towards the important role of both static and dynamic stimuli in the growth of embryonic limbs and the development of a functional joint space. PMID- 15758414 TI - Muscle power, locomotor performance and flexibility in aging mentally-retarded adults with and without Down's syndrome. AB - Longer life expectancy is resulting in increasing numbers of elderly adults with mental retardation (MR). The objective of the study was to compare lower limb isokinetic muscle power, locomotor performance and flexibility of aged adult mentally-retarded individuals with and without Down's syndrome (DS). Nine subjects with MR and DS (mean age 61), and sixteen subjects with MR and without DS (mean age 63), performed leg power testing on a Biodex dynamometer. Parameters measured were dynamic torque, dynamic torque % body weight, and average power % body weight. Functional performance tests including "Timed Get-Up and Go" and flexibility were also analyzed and compared. Results indicate that in knee extension and flexion isokinetic power the MR group without DS showed significantly higher scores than the MR group with DS. The functional performance of elderly adults with MR and DS was significantly impaired compared with MR adults without DS, although no differences were observed between the two groups in the flexibility tests. It was concluded that muscle leg power, and gross motor performance of elderly mentally-retarded individuals without Down's syndrome is better than in those with Down's syndrome. PMID- 15758415 TI - Bone mineral density in beta - thalassemic Lebanese children. AB - Osteoporosis has been described extensively in adult thalassemics. Fewer studies have been reported in thalassemic children. In this article, we measured the bone mineral density (BMD) of Lebanese beta-thalassemic children before institution of a balanced transfusion-chelation regimen, in comparison with that of healthy controls, and studied its correlation with various demographic and biological parameters. Both groups, controls and thalassemics were comparable with respect to age, sex, socioeconomic and regional distribution. On the other hand, thalassemics had a significantly lower height age (p<0.001), lower bone age (p=0.001), lower sexual maturation (p=0.004), lower absolute BMD values and larger negative BMD-Z scores (p<0.001). Within the thalassemic group, BMD correlated significantly with luteinizing hormone (LH) and follicle stimulating hormone (FSH), estradiol and testosterone values, as well as with the pretransfusion hematocrit, but not with other endocrine or bone metabolism parameters. We conclude that Lebanese beta-thalassemic children have a significantly lower BMD than their healthy counterparts due, in part, to their slower physical development. A major contributor seems to be the low-transfusion regimen followed by these patients, as well as the endocrine dysfunction which was detected in about 25% of them. PMID- 15758416 TI - Effects of nicotine on bone and calciotropic hormones in aged ovariectomized rats. AB - The objective of this investigation was to assess the effects of chronic nicotine administration on bone status and serum calcium and calciotropic hormone levels in aged, estrogen-replete (intact, sham-operated) and estrogen-deplete (ovariectomized) female rats. Eight-month-old sham-operated (sham) and ovariectomized (ovx) retired breeder rats were maintained untreated for 3 months to allow for the development of osteopenia in the ovx group. The animals were then administered either saline, low dose nicotine (6.0 mg/kg/day), or high dose nicotine (9.0 mg/kg/day) via osmotic minipumps for 3 months. Blood was drawn at necropsy for determination of serum nicotine, cotinine, Ca, PTH, 25(OH)D, and 1,25(OH)(2)D. Right tibiae were collected and processed undecalcified for cancellous and cortical bone histomorphometry. Histomorphometric endpoints evaluated at the proximal tibial metaphysis included cancellous bone volume (BV/TV), osteoclast surface (Oc.S), osteoid surface (OS), mineralizing surface (MS), mineral apposition rate (MAR), and bone formation rate (BFR). Histomorphometric endpoints evaluated at the tibial diaphysis included cortical area (Ct.Ar), marrow area (Ma.Ar), and periosteal and endocortical MS, MAR, and BFR. Ovariectomy resulted in lower cancellous BV/TV and Ct.Ar and higher cancellous, endocortical, and periosteal MS and BFR. The presence of nicotine in serum confirmed successful delivery of the drug via osmotic minipumps. Administration of nicotine at the high dose resulted in lower serum 25(OH)D levels but differences in serum Ca or PTH were not detected with either nicotine treatment. Differences with nicotine treatment were also not detected for Oc.S at the proximal tibia. While treatment with nicotine at the high dose resulted in higher MS and BFR, in both sham and ovx rats, there were no differences due to nicotine treatment in cancellous BV/TV. Marrow area was greater in rats treated with nicotine than in rats treated with vehicle. However, differences with nicotine treatment were not detected in Ct.Ar in either intact or ovx rats. Overall, these findings indicate that steady state nicotine exposure does not alter bone mass in intact or ovx rats but may have detrimental effects on body storage of vitamin D. PMID- 15758418 TI - What is new in neuro-musculoskeletal interactions? PMID- 15758417 TI - The role of estrogen receptor-beta, in the early age-related bone gain and later age-related bone loss in female mice. AB - The molecular and cellular mechanism of estrogen action in skeletal tissue remains unclear. The purpose of this study was to understand the role of estrogen receptor-beta, (ERbeta) on cortical and cancellous bone during growth and aging by comparing the bone phenotype of 6- and 13-month-old female mice with or without ERbeta. Groups of 11-14 wild-type (WT) controls and ERbeta knockout (BERKO) female mice were necropsied at 6 and 13 months of age. At both ages, BERKO mice did not differ significantly from WT controls in uterine weight and uterine epithelial thickness, indicating that ERbeta does not regulate the growth of uterine tissue. Femoral length increased significantly by 5.5% at 6 months of age in BERKO mice compared with WT controls. At 6 months of age, peripheral quantitative computerized tomography (pQCT) analysis of the distal femoral metaphysis (DFM) and femoral shafts showed that BERKO mice had significantly higher cortical bone content and periosteal circumference as compared with WT controls at both sites. In contrast to the findings in cortical bone, at 6 months of age, there was no difference between BERKO and WT mice in trabecular density, trabecular bone volume (TBV), or formation and resorption indices at the DFM. In 13-month-old WT mice, TBV (-41%), trabecular density (-27%) and cortical thickness decreased significantly. while marrow cavity and endocortical circumference increased significantly compared with 6-month-old WT mice. These age-related decreases in cancellous and endocortical bone did not occur in BERKO mice. At 13 months of age, BERKO mice had significantly higher total, trabecular and cortical bone, while having significantly lower bone resorption, bone formation and bone turnover in DFM compared with WT mice. These results indicate that deleting ERbeta protected against age-related bone loss in both the cancellous and endocortical compartments by decreasing bone resorption and bone turnover in aged female mice. These data demonstrate that in female mice, ERbeta plays a role in inhibiting periosteal bone formation, longitudinal and radial bone growth during the growth period, while it plays a role in stimulating bone resorption, bone turnover and bone loss on cancellous and endocortical bone surfaces during the aging process. PMID- 15758419 TI - The 3rd ISMNI Workshop. Introduction. PMID- 15758420 TI - Volumetric and projective bone mineral density. AB - Projectional bone mineral density measurement so far used extensively in radiogrammetry and single and dual source absorptiometry is confronted with a serious limitation for the accurate evaluation of true density artifactually providing higher values along with the increase of body size and bone depth on account of the omission of one dimension. Computed tomography is capable of measuring true volumetric density and also accomplishes a separate measurement of trabecular and cortical bone especially on application to the distal and mid radius with abundant cortical bone in peripheral computed tomography (pQCT). New lines of information may be obtained by such separate trabecular and cortical bone measurement in decreases of bone density due to various causes, estrogen withdrawal, corticosteroid, diabetes mellitus, renal failure, etc. Dynamic analysis of the result of pQCT may also make it possible to assess bone strength and resistance to fracture. PMID- 15758421 TI - The bone behind a low areal bone mineral density: peripheral quantitative computed tomographic analysis in a woman with osteogenesis imperfecta. AB - Areal bone mineral density (BMD) is the most widely used densitometric parameter. However, this approach makes it difficult to understand the structural basis of bone diseases, because a large number of bone properties are integrated into a single number. This is exemplified in the present case of a 27-year-old woman with osteogenesis imperfecta type I. Peripheral quantitative computed tomographic analysis at the radial metaphysis and at the radial diaphysis revealed a decreased areal BMD at both sites (z score -3.9 and -3.4, respectively). Yet, the structural basis for this decrease was different for the two locations: At the distal radius areal BMD was decreased because volumetric BMD was very low, whereas bone size was above the mean of the reference range. At the proximal radius areal BMD was decreased, because bone size was very low but volumetric BMD was above average. Bone mineral content of the radial diaphysis was very low for forearm muscle size, a finding which is compatible with Frost's hypothesis that the mechanostat setpoint is increased in osteogenesis imperfecta. PMID- 15758422 TI - Adult mesenchymal stem cells: potential for muscle and tendon regeneration and use in gene therapy. AB - The expansion potential and plasticity of stem cells, adult or embryonic, offer great promise for their use in medical therapies. Recent provocative data suggest that the differentiation potential of adult stem cells may extend to lineages beyond those usually associated with the germ layer of origin. In this review, we describe recent developments related to adult stem cell research and in particular, in the arena of mesenchymal stem cell (MSC) research. Research demonstrates that transduced MSCs injected into skeletal muscle can persist and express secreted gene products. The ability of the MSC to differentiate into cardiomyocytes has been reported and their ability to engraft and modify the pathology in infarcted animal models is of great interest. Research using MSCs in tendon repair provides information on the effects of physical forces on phenotype and gene expression. In turn, MSCs produce changes in their matrix environment in response to those biomechanical forces. Recent data support the potential of MSCs to repair tendon, ligament, meniscus and other connective tissues. Therapeutic applications of adult stem cells are approaching clinical use in several fields, furthering the possibility to regenerate damaged and diseased tissue. PMID- 15758423 TI - The role of strain in the response of rapidly growing young male rat bones to parathyroid hormone. AB - Human parathyroid hormone (hPTH 1-34) stimulates an anabolic response in human and animal skeletons; however, it is unclear if the effect is strain dependent. To determine if the anabolic response to hPTH (1-34) was dependent upon strain in rats we used 2 outbred strains (Sprague Dawley, Wistar), 2 inbred strains (Fischer 344, Wistar spontaneously hypertensive:SHR), and 2 mutant strains (Zucker obese, Zucker lean) of rats. Male rats, 5 weeks of age, from each strain were treated subcutaneously with 80 microg/kg body weight hPTH (1-34) or vehicle for 12 days. The response to PTH was similar in all strains whereby PTH exerted an anabolic effect on femoral bone mass and cancellous bone histology that was independent of strain differences. Histomorphometric indices of bone volume, mineralized surface and bone formation in lumbar vertebrae increased in all PTH treated rats. Additionally, femur bone mineral content and bone mineral density measured by dual energy X-ray absorptiometry (DEXA), and ash weight increased in all PTH-treated rats. These increases occurred regardless of strain. In summary, PTH exerted comparable anabolic effects on bone mass, bone mineral density and bone formation in all rat models tested demonstrating that the skeletal responsiveness to PTH was not dependent upon strain. PMID- 15758424 TI - Nerve growth factor and norepinephrine concentrations in weight-bearing and non weight-bearing bones of euthyroid and hyperthyroid rats. AB - Thyroid hormone, nerve growth factor (NGF) and norepinephrine (NE) and weight bearing affect bone metabolism, yet interactions between these factors and osseous tissue have not been investigated. Therefore, the aims of the study were to measure NGF and NE concentrations in weight-bearing and non-weight-bearing bones from euthyroid (control) and hyperthyroid (HT) rats. Hyperthyroidism was induced by oral intake of triiodothyronine (90 mg/kg/day) for 21 days. Histomorphometry on distal femurs verified significant trabecular bone loss in HT rats compared to euthyroid animals. NGF concentrations were assayed via ELISA, whilst NE concentrations were measured via HPLC and ECD. In euthyroid rats: (i) the concentration of NGF in ribs (914 ng/g) was almost 3-fold greater than in femurs (326 ng/g wet weight of tissue) (ii) the concentrations of NE in ribs (74.7 ng/g) and calvaria (87.4 ng/g) were 2.5-3.5-fold greater than either femurs (24.0 ng/g) or tibiae (30.5 ng/g) and (iii) NE concentrations were comparable between ribs (74.7 ng/g) and calvaria (87.4 ng/g) and similar between tibiae (30.5 ng/g) and femurs (24.0 ng/g). In HT rats: (i) the concentration of NGF in ribs (1802 ng/g) was 4-fold greater than in femurs (402 ng/g) (ii) NE concentrations in ribs (23.3 ng/g) and calvaria (13.6 ng/g) were 4.5-fold and 2.6 fold greater respectively than in tibiae (5.2 ng/g), while ribs had almost a 2 fold higher concentration of NE than calvaria. In HT rats compared to euthyroid animals: (i) NGF concentrations almost doubled in ribs but there was little change in the NGF concentration in femurs (ii) there was a reduction in NE concentrations in calvaria by 84%, in ribs by 69% in tibiae by 83% and 55% in femur (NS). CONCLUSIONS: (i) Non-weight-bearing is associated with higher concentrations of NGF and NE than weight-bearing in bones in euthyroid and HT rats; (ii) Hyperthyroidism exerts opposite effects on NGF and NE in bone and (iii) Hyperthyroidism interacts with weight-bearing to determine NGF and NE concentrations in bone. Therefore, the influence of thyroid hormone on NGF and NE in bone may need to be taken into account when considering the action of thyroid hormone on bone in either euthyroid or hyperthyroid states. PMID- 15758425 TI - Alendronate as an effective countermeasure to disuse induced bone loss. AB - Microgravity, similar to disuse immobilization on earth, causes rapid bone loss. This loss is believed to be an adaptive response to the reduced musculoskeletal forces in space and occurs gradually enough that changes occurring during short duration space flight are not a concern. Bone loss, however, will be a major impediment for long duration missions if effective countermeasures are not developed and implemented. Bed rest is used to simulate the reduced mechanical forces in humans and was used to test the hypothesis that oral alendronate would reduce the effects of long duration (17 weeks) inactivity on bone. Eight male subjects were given daily oral doses of alendronate during 17 weeks of horizontal bed rest and compared with 13 male control subjects not given the drug. Efficacy was evaluated based on measurements of bone markers, calcium balance and bone density performed before, during and after the bed rest. The results show that oral alendronate attenuates most of the characteristic changes in bone that are associated with long duration bed rest and presumably space flight. PMID- 15758426 TI - Early closure of growth plate causes poor growth of long bones in collagen induced arthritis rats. AB - Abnormalities of the epiphyseal growth plate that occur in collagen-induced arthritis (CIA) were studied. CIA was induced in 6-week-old Lewis rats by immunization with type II collagen. Radiographic examination revealed the early closure of the epiphyseal growth plate with growth retardation of the femur and tibia. Histological evaluation confirmed the early closure of the epiphyseal growth plate accompanied by decreased intensity of safranin-O staining indicating decreased amounts of proteoglycans in the extracellular matrix (ECM) of the cartilage. Immunohistochemical methods showed that the number of chondrocytes expressing matrix metalloproteinase (MMP)-3 and/or vascular endothelial growth factor (VEGF) increased in the growth plates of CIA rats. This study confirmed that disturbances of long bone growth with early closure of the epiphyseal growth plates occur in CIA. There appeared to be overexpression of MMP-3, which may be involved with proteoglycan degradation. Additionally, VEGF, which is associated with cartilage ossification and angiogenesis, might also play a role in this event. Further clarification of the mechanism of the growth disturbance in CIA may yield clinical benefits, especially in prevention of the premature closure of growth plate that is seen in juvenile rheumatoid arthritis and other diseases. PMID- 15758429 TI - The 31st Sun Valley Hard Tissue Workshop. Introduction. PMID- 15758427 TI - Isoflavones and bone: animal and human evidence of efficacy. AB - Previous reports of soy extracts and isoflavone-enriched preparations studied in animals and humans have found that these molecules, when given at appropriate doses, have positive effects on the skeleton, including improvements in bone mineral content (BMC) and bone mineral density (BMD). A reduction in fracture risk of human subjects has not yet been shown in a prospective trial. Isoflavones, which exist in significant amounts only in soybeans, exert estrogen like effects in human bone cells because of their unique organic structures that are similar to that of estradiol. The discovery of the b isoform of the estrogen receptor (ER) suggests that the molecular regulation of bone remodeling by estrogens, or estrogen-like molecules, including isoflavones, is more complex than previously thought. Depending on the type of ER present in a particular tissue, isoflavones may act as weak estrogen agonists or as weak estrogen antagonists. For example, isoflavones act as weak estrogen agonists in osteoblasts, but in reproductive cells, such as in the breast and uterus, they behave as weak estrogen antagonists. Weak agonistic effects of the isoflavones include stimulation of osteoblast proliferation and differentiation and increasing the production of cytokines that may inhibit osteclastic activity. The selective beneficial effects of estrogen-like molecules in bone tissues, compared to the anti-estrogenic effects in cells of reproductive tissues, make isoflavones attractive for the promotion of bone health. Relatively greater values of BMC and BMD of Asian populations with high consumption of soy isoflavones throughout life, compared to those with lower intakes, indirectly support the skeletal benefits of this pattern of intake of these estrogen-like molecules. PMID- 15758430 TI - Osteoporosis in women. PMID- 15758431 TI - An expanded overview of postmenopausal osteoporosis. AB - Why is the incidence of osteoporotic fracture so much higher in women than in men? The dominant medical view holds that the exaggerated skeletal fragility and fracture risk of postmenopausal women solely reflects the loss of bone following withdrawal of endogenous estrogen. Indeed, an enormous amount of research in this area has attempted to understand the rise in fractures after menopause in terms of the impact of estrogen lack on bone remodeling. Recent insights suggest that this simple view does not offer an adequate explanation for the greater susceptibility of older women to fracture compared to that of men. It seems more reasonable to view bone health as a lifelong process, reflecting the contributions and influences of myriad events occurring throughout life to skeletal acquisition and maintenance. Only recently has the medical community recognized that the amount of bone present at skeletal maturity makes a powerful contribution to lifelong skeletal status. A second area that must be incorporated into discussions of this topic relates to bone size and geometry. Women's bones are inherently smaller than those of men. A bone's strength is determined by its size as well as by its material properties. In boys, pubertal increases in the cortical thickness of long bones are achieved by (testosterone-dependent) periosteal apposition. By contrast, increased cortical thickness in girls reflects bone expansion into the medullary space, with little or no periosteal apposition, suggesting an inhibitory effect of estrogen on the latter process. Consequently, at skeletal maturity, men have wider bones of greater mechanical competence. Although estrogen is generally held to be skeletally protective, this aspect of its actions may actually render women more susceptible to some fractures. In later life, men may lose even more bone from appendicular sites than do women, but men show much greater concomitant increases in periosteal apposition than women, permitting them to maintain a relatively favorable mechanical profile. These several findings are based on cross-sectional observations of relatively few individuals and therefore require confirmation in prospective longitudinal studies. The degree to which gender-related differences in later life skeletal adaptation reflects a bone's mechanical or metabolic environment has been frequently discussed but still awaits experimental confirmation. PMID- 15758432 TI - How drugs decrease fracture risk: lessons from trials. AB - In women with osteoporosis, each 1% improvement in spine BMD (by DXA) is expected to reduce vertebral fracture risk by about 4%. However, randomized trials of antiresorptive agents show that 1 to 6% improvements in spine BMD reduce vertebral fracture risk by 35 to 50%. Less 20% of the decreased spine fracture risk produced by alendronate or raloxifene be explained by improvement in spine BMD. The discrepancy is even greater during the first year or two of treatment when 1 to 4% improvements in BMD are associated with 65-68% decreases in spine fracture risk. Bisphosphonates continue to increase BMD but the reduction in fracture risk wanes to 20 to 45%. DXA underestimates the change in bone density of spinal trabecular bone and this might explain part of the discrepancy between expected and observed reductions in spine fracture risk. Even more accurate measurement of BMD would not explain the rapid onset and later waning of effect despite gradually increasing BMD. The biomechanical effects inhibiting bone resorption could explain the early onset but not the waning effectiveness. The waning effectiveness of antiresorptives raises concerns that prolonged inhibition of remodeling may weaken bone by allowing microdamage to accumulate. The effect of drugs on nonspine fracture risk is more complex and cannot be predicted from changes in DXA BMD. For example, Beck showed that long-term users of estrogen increase section modulus vs. nonusers with a net increase in section modulus and predicted femoral neck strength despite losing about 0.4% per year in femoral neck BMD. PTH reduces spine fracture risk and this effect is more completely explained by improvement in spine BMD. This suggests that sustaining the increased BMD produced by PTH may maintain long-term reductions in fracture risk. PMID- 15758433 TI - Bone material properties and mineral matrix contributions to fracture risk or age in women and men. AB - The strength of bone is related to its mass and geometry, but also to the physical properties of the tissue itself. Bone tissue is composed primarily of collagen and mineral, each of which changes with age, and each of which can be affected by pharmaceutical treatments designed to prevent or reverse the loss of bone. With age, there is a decrease in collagen content, which is associated with an increased mean tissue mineralization, but there is no difference in cross-link levels compared to younger adult bone. In osteoporosis, however, there is a decrease in the reducible collagen cross-links without an alteration in collagen concentration; this would tend to increase bone fragility. In older people, the mean tissue age (MTA) increases, causing the tissue to become more highly mineralized. The increased bone turnover following menopause may reduce global MTA, and would reduce overall tissue mineralization. Bone strength and toughness are positively correlated to bone mineral content, but when bone tissue becomes too highly mineralized, it tends to become brittle. This reduces its toughness, and makes it more prone to fracture from repeated loads and accumulated microcracking. Most approved pharmaceutical treatments for osteoporosis suppress bone turnover, increasing MTA and mineralization of the tissue. This might have either or both of two effects. It could increase bone volume from refilling of the remodeling space, reducing the risk for fracture. Alternatively, the increased MTA could increase the propensity to develop microcracks, and reduce the toughness of bone, making it more likely to fracture. There may also be changes in the morphology of the mineral crystals that could affect the homogeneity of the tissue and impact mechanical properties. These changes might have large positive or negative effects on fracture incidence, and could contribute to the paradox that both large and small increases in density have about the same effect on fracture risk. Bone mineral density measured by DXA does not discriminate between density differences caused by volume changes, and those caused by changes in mineralization. As such, it does not entirely reflect material property changes in aging or osteoporotic bone that contribute to bone's risk for fracture. PMID- 15758434 TI - Architecture and trabecular bone - toward an improved understanding of the biomechanical effects of age, sex and osteoporosis. AB - From an engineering perspective, trabecular bone is a highly complex material, being anisotropic with different strengths in tension, compression, and shear and with mechanical properties that vary widely across anatomic sites, and with aging and disease. While mechanical properties depend very much on volume fraction, the role of architecture and tissue material properties remain uncertain. In the context of osteoporosis, there is wide interest in the biomechanical role of architecture since this should lead to improved understanding of the disease and ultimately better diagnosis and drug treatment assessment. This study reviews what is known about architectural changes in trabecular bone associated with age, gender and osteoporosis and the role of these changes in the mechanical properties of bone. Recent development of three-dimensional high-resolution imaging technologies has provided more accurate measures of quantitative metrics of architecture, thereby providing new data and raising questions about earlier conclusions. Focusing on the hip and spine, this literature is synthesized and outstanding issues are identified. In addition, the changing paradigm of biomechanical research on trabecular architecture is addressed. Because of the complexity of the trabecular micromechanics, the prevailing approach to date can be classified as an inverse one, whereby candidate metrics of architecture are developed and tested for efficacy in an empirical trial-and-error fashion. In this approach, the biomechanics is treated only as an assay since it is not used to guide development of the candidate metrics. By contrast, a more forward approach is to study the associated micromechanics using engineering analysis and from that identify the metrics that in theory most affect mechanical properties. The latter approach, facilitated by the new high-resolution imaging techniques and increased computational power, is discussed in an attempt to direct attention to new types of architectural metrics that are independent of bone density and that should improve the ability to explain how age, gender and osteoporosis affect the mechanical properties of trabecular bone. PMID- 15758435 TI - Size, structure and gender: lessons about fracture risk. AB - The differences in age-related fracture risks among men and women must reflect gender differences in the relevant variables. We are concerned here with gender differences in structural variables that relate to the size and shape of bones. As children grow, their bones grow in diameter through periosteal modeling. Studies show that radial growth is driven by mechanical forces and is not just "genetically programmed". Moving bone mass farther from the center of the diaphysis makes it more effective in resisting bending and twisting forces, and disproportionately so in comparison to changes in bone mass. Gender differences in long bone structure appear to arise because the bone cells of males and females function in different hormonal environments which affect their responses to mechanical loading. In girls, bone formation on the metacarpal periosteal surface essentially stops at puberty, and is replaced by formation on the endosteal surface, reducing endosteal diameter until about age 20. Bone strength is 60% greater in male metacarpals than in those of females because bone is added periosteally in boys and endosteally in girls. At menopause endosteal resorption resumes, accompanied by slow periosteal apposition, weakening cortical structure. Similar phenomena occur in such critical regions as the femoral neck. Another fundamental gender difference in skeletal development is that whole body bone mineral content increases in linear proportion to lean body mass throughout skeletal maturation in boys, but in girls there is a distinct increase in the slope of this relationship at puberty, when estrogen rises. Frost's hypothesis is that this reflects an effect of estrogen on bone's mechanostat set point, and this is increasingly supported by data showing that estrogen and mechanical strain act through a common pathway in osteoblast-like cells. If Frost's hypothesis is correct, the mechanostat is set for maximal effect of mechanical loading on bone gain during the 2-3 years preceding menarche. During the childbearing years, the set point is at an intermediate level, and at menopause, it shifts again to place the skeleton into the metabolic equivalent of a disuse state. The most direct approach to resolving this problem would be to simulate the putative effect of estrogen on the set point itself. PMID- 15758436 TI - Osteopenic mice: animal models of the aging skeleton. AB - While our understanding of the developmental biology of the skeleton, like that of virtually every other subject in biology, has been transformed by recent advances in human and mouse genetics, we still know very little, in molecular and genetic terms, about skeletal physiology. Thus, among the many questions that are largely unexplained are the following: why is osteoporosis mainly a women's disease? How is bone mass maintained nearly constant between the end of puberty and the arrest of gonadal functions? Molecular genetics has emerged as a powerful tool to study previously unexplored aspects of the physiology of the skeleton. Among mammals, mice are the most promising animals for this experimental work. This has been previously demonstrated e.g. through the tremendous impact of the different osteopetrotic models on our molecular understanding of osteoclastic bone resorption. Until recently the only way of studying bone loss situations and osteoporosis in mice was by using ovariectomy with all its limitations. Today, however, we have access to more sophisticated osteoporotic mouse-models from four different origins: Transgenic mice (HSV-TK), knock-out mice (OPG), inbred-strains (SAMP6), and through physiological modulation (icv application). These new models have already taught us several important lessons. The first is, that bone remodeling is more than just an autocrine/paracrine process. Multiple experimental evidence has demonstrated that the latter regulation exists, but genetics prove that there is no functional cross-control between resorption and formation. The second lesson is, that remodeling is, at least in part, subject to central regulation. Thus, osteoporosis is partly a central or hypothalamic disease. However, the most dramatic change and the most important advantage we feel is, that today we have models to test a new hypothesis regarding the etiology of osteoporosis before it turns to dogma. Taken together, mouse-studies may lead to a shift in our physiological understanding of skeleton biology and to the emergence of novel paradigms. These, in turn, should help us to devise new treatments for degenerative diseases of the skeleton such as osteoporosis and its associated clinical problems. PMID- 15758437 TI - Osteoporosis in men. PMID- 15758438 TI - The determinants of fracture in men. AB - Osteoporosis represents an increasingly important clinical and public health problem among older men. Estimates indicated that 1-2 million (3-6%) men aged 50 years and over in the United States have osteoporosis and 8-13 million (28- 47%) have osteopenia. The lifetime risk of suffering a hip, spine or forearm fracture for a 50-year-old man is 13%, similar to the risk for prostate cancer. The number of osteoporotic fractures in men is expected to increase dramatically due to aging of the population and secular increases in fracture rates. Identification of men who are at greatest risk of osteoporosis and the risk factors, which predispose men to fracture, are essential so that preventive steps can be taken. Data on risk factors are emerging but many questions remain. Men may fracture at a higher bone mineral density (BMD) level than women. However, estimates of volumetric BMD, which correct in part for gender differences in bone size, and risk of fracture, may actually show similar relationships in men and women. Fracture rates are similar in older African American women and Caucasian men. Improved understanding of ethnic differences in fracture could identify potential reasons for gender differences. Family history and genetic factors are also important risk factors for fractures but the specific candidate genes are not known and whether gender modifies the effects of these genetic polymorphisms on BMD and the risk of fracture is also not known. In general, lifestyle factors and anthropometric measurements show similar relationships with fractures in men and women although few comprehensive prospective studies have been conducted. Current data will be reviewed on the relationships between markers of skeletal health, genetic polymorphisms, lifestyle and anthropometric factors and fracture. PMID- 15758439 TI - Novel, non-steroidal, selective androgen receptor modulators (SARMs) with anabolic activity in bone and muscle and improved safety profile. AB - A novel approach to the treatment of osteoporosis in men, and possibly women, is the development of selective androgen receptor modulators (SARMs) that can stimulate formation of new bone with substantially diminished proliferative activity in the prostate, as well as reduced virilizing activity in women. Over the last several years, we have developed a program to discover and develop novel, non-steroidal, orally-active selective androgen receptor modulators (SARMs) that provide improved therapeutic benefits and reduce risk and side effects. In recent studies, we have used a skeletally mature orchiectomized (ORX) male rat as an animal model of male hypogonadism for assessing the efficacy of LGD2226, a nonsteroidal, non-aromatizable, and non-5alpha-reducible SARM. We assessed the activity of LGD2226 on bone turnover, bone mass and bone strength, and also evaluated the effects exerted on classic androgen-dependent targets, such as prostate, seminal vesicles and muscle. A substantial loss of bone density was observed in ORX animals, and this loss was prevented by SARMs, as well as standard androgens. Biochemical markers of bone turnover revealed an early increase of bone resorption in androgen-deficient rats that was repressed in ORX animals treated with the oral SARM, LGD2226, during a 4-month treatment period. Differences in architectural properties and bone strength were detected by histomorphometric and mechanical analyses, demonstrating beneficial effects of LGD2226 on bone quality in androgen-deficient rats. Histomorphometric analysis of cortical bone revealed distinct anabolic activity of LGD2226 in periosteal bone. LGD2226 was able to prevent bone loss and maintain bone quality in ORX rats by stimulating bone formation, while also inhibiting bone turnover. LGD2226 also exerted anabolic activity on the levator ani muscle. Taken together, these results suggest that orally-active, non-steroidal SARMs may be useful therapeutics for both muscle and bone in elderly hypogonadal men through their anabolic activities. Since SARMs both prevent bone loss, and also stimulate formation of new bone, they may have significant advantages relative to currently used anti-resorptive therapies. Coupled with their activity in muscle and their ability to maintain or restore libido, they offer new therapeutic approaches for male and female hormone replacement. PMID- 15758440 TI - Genetics and bone. Using the mouse to understand man. AB - The rationale for use of inbred strains of mice in bone research is well recognized and includes: a) practical factors (economics of scale, rapid development of adult status, pre-existing knowledge, down-sized technologies) and b) proven methodologies for genetic studies (polygenic trait analyses, mapping tools, genomic sequencing, methods for gene manipulation). Initial investigations of inbred strains of mice showed that femoral and lumbar vertebral volumetric bone mineral density (BMD, mg/mm(3)) by pQCT varied in excess of 50% for femurs and 9% in vertebral BMD. Two strains - low BMD C57BL/6J (B6) mice and high BMD C3H/HeJ (C3H) - were investigated for insights to their BMD diversity. B6C3F2 females derived from intercrossing B6C3F1s were raised to adult skeletal status at 4 months, then necropsied for phenotyping of bone and genotyping of genomic DNA. 1000 F2 females were genotyped for PCR product polymorphisms on all 19 autosomes at approximately 15 cM. Genome wide analyses for genotype-phenotype correlations showed 10 chromosomes (Chrs) carried genes for femoral and 7 Chrs for vertebral BMD. LOD scores ranged from 2.90 to 24.4, and percent of F2 variance accounted for ranged from 1 to 10%. Analyses of main effects revealed both dominant-recessive and additive inheritance patterns. Both progenitor strains carried alleles with positive and negative effects on BMD of each bone sites. A remarkable array of additonal skeletal phenotypes (femur and vertebral geometry, strength measures, serum markers) also proved polygenic in nature, with complex segregation patterns. Verification of BMD quantitative trait loci (QTLs) was undertaken by creating congenic B6 strains carrying individual QTL regions from C3H. Following 6 cycles of backcrossing a QTL-containing region from C3H to the B6 strain, N6F2 congenic strain mice were aged to 4 months, then genotyped for the QTL region and phenotyped for skeletal traits. Comparison of mice homozygous for C3H alleles versus homozygous for B6 alleles in the QTL regions showed that femoral BMD increased or decreased significantly in congenic strains, as was predicted from F2 data. Gender differences specific to BMD QTLs have been revealed, as have more than 30 additional phenotypes associated with cortical and trabecular structural parameters and biomechanical properties. PMID- 15758441 TI - Genetic regulation of bone mineral density in mice. AB - Peak bone mass is a major determinant of risk of osteoporotic fracture. Family and twin studies have found a strong genetic component to the determination of bone mineral density (BMD). However, BMD is a complex trait whose expression is confounded by environmental influences and polygenic inheritance. The number, locations and effects of the individual genes contributing to natural variation in this trait are all unknown. The extreme difficulty of dissecting out environmental factors from genetic ones in humans has motivated the investigation of animal models. Genetically distinct animal strains raised under strict environmental control are critical tools for defining genetic regulation. The availability of inbred strains, combined with its relative fecundity, has established the mouse as the best model system for the study of mammalian genetics and physiology. Importantly, genes identified in murine analyses can usually be readily mapped to particular human chromosomal regions because of the high degree of synteny that exists between the mouse and human genomes. We employed quantitative trait locus (QTL) analysis to examine peak BMD in 24 recombinant inbred (RI) mouse strains, derived from a cross between C57BL/6 (B6) and DBA/2 (D2) progenitors (BXD RI). The distribution of BMD values among these strains clearly indicated the presence of strong genetic influences, with an estimated narrow sense heritability of 35%. The differences in peak whole body BMD in the BXD strains were integrated with a large database of genetic markers previously defined in the RI BXD strains to generate chromosome map sites for QTL locations. This QTL analysis provisionally identified a number of chromosomal sites linked to BMD. In the second phase of our BMD QTL mapping efforts, we used three independent mouse populations (all derived from B6 and D2 progenitor strains) to confirm and narrow the genetic locations of 4 QTLs (on chromosomes 1, 2, 4, and 11) that strongly influence the acquisition of peak BMD in mice. Using a novel, fine-mapping approach (recombinant inbred segregation testing), we have succeeded in narrowing two of the BMD-related chromosomal regions and in the process eliminated a number of candidate genes. The homologous regions in the human genome for each of these murine QTLs have been identified in recent human genetic studies. In light of this, we believe that findings in mice should aid in the identification of specific candidate genes for study in humans. PMID- 15758442 TI - Is skeletal mechanotransduction under genetic control? AB - Studies of twins have established that peak bone mass is about 70% heritable. The skeletal response to exercise contributes to peak bone mass, as mechanical loading increases skeletal mass during growth and development. It is possible that the skeletal responsiveness to mechanical loading is under genetic control, so that some individuals will build stronger bones with exercise. This appears to be the case in mice. Long bones in mice of the C3H/He strain are largely unresponsive to mechanical loading. Ironically, this strain of mice has very high bone density. Perhaps the genes that regulate BMD are not the same as those that regulate mechanical loading response. Studies of recombinant inbred and congenic strains derived from C3H mice will help to identify genes influencing bone size, density and responsiveness to mechanical loading. PMID- 15758443 TI - Osteocytes: a proposed multifunctional bone cell. AB - Most cell types are ascribed a single function. The osteoclast holds the unique distinction of performing only one function in the body - that of resorbing bone. The osteoblast has been ascribed the major function of bone matrix production. Other less well-defined cell types include progenitor cells and the nebulous cell type that can support osteoclast formation upon stimulation with various bone resorbing cytokines. Obviously, these cells could have other functions. The definition of an osteocyte is descriptive of its location - cells surrounded by mineralized matrix - not its function. For this year's Sun Valley Workshop on osteocytes, several proposed functions will be presented. First, a general consensus exists that osteocytes are most likely sensitive to mechanotransduction and translate mechanical strain into biochemical signals. Consensus does not exist on the nature of the mechanical strain, the form of the biochemical signals, the target cell(s), or the viability status of the osteocyte. Second, it is also proposed that this cell is incredibly adaptable and expresses plasticity in response to mechanical stimuli. In other words, this cell can readjust its responses to strain in the presence of other bone agents such as hormones and bone factors. Third, it will also be presented that osteocytes maintain systemic mineral homeostasis by regulating mineral release and deposition over the enormous surface area over which these cells interface with the surrounding matrix. Although osteocytes are terminally differentiated osteoblasts, they appear to have separate and distinct properties from their predecessors. Bone cell biologists loaded with an arsenal of bone anabolic and catabolic factors are examining the expression and effects of these factors on osteocytes. Engineers trained in mathematical modeling have generated new models of strain and connectivity to be tested. The unique morphology of osteocytes suggests that the cytoskeleton in these cells may function differently from osteoblasts and other cell types. Osteocytes may consist of different subpopulations; some that possess receptors for parathyroid hormone (PTH) and others that only express receptors for carboxyl terminal PTH suggesting different functions and responses. Osteocytes may respond rapidly to strain through glutamate receptor-like mechanisms, through calcium influxes, through gap junctions, and less rapidly through the production of small molecules and factors. Strain may take the form of substrate stretching and/or fluid flow. Osteocytes may communicate with other osteocytes and/or bone surface cells such as lining cells, stromal cells, osteoblasts, and/or osteoclasts and their precursors. The viability status of the osteocyte may determine the type of signals sent from these cells. If the cells are deprived of oxygen or nutrients, the apoptotic cells may send signals for initiation of resorption. If the cells and/or their dendritic process are ripped or torn by microdamage, they may send signals of both resorption and formation. If the majority of these theories are correct, then the osteocyte is the 'smart' cell that can direct or orchestrate the bone resorbing and bone forming cells even in its death and dying. PMID- 15758444 TI - The role of osteocytes in bone regulation: mineral homeostasis versus mechanoreception. AB - Early work on the role of osteocytes in bone regulation suggested that the primary function of these cells was osteolysis. This lytic function was not precisely defined but included mineral homeostasis and at least the initiation of matrix remodeling, if not a primary role in remodeling. This paper is an attempt to promote the concept of osteocytic osteolysis as a method of systemic mineral homeostasis and to separate it from bone remodeling. Although recent investigations have pointed to mechanotransduction as a primary function of osteocytes, resulting in a general abandonment of the osteocytic osteolysis concept, the corpus of evidence suggests that osteocytes likely have a multipurpose role in the biology of bone. The osteocyte network represents an enormous surface area over which the cells interface with the surrounding matrix, useful for both strain detection and matrix mineral access. Osteocytes have been found to possess receptors for PTH, a known regulator of mineral ion homeostasis. Cultured osteocytes placed on dentin slices demonstrated no capacity to pit the dentin, but they were not treated with a regulating factor such as PTH, nor does mineral homeostasis require substantial bone volume removal. Scaling relationships suggest that osteocyte density is inversely proportional to body mass, R(2) = 0.86, and thus directly proportional to metabolic rate. Thus, species with higher metabolic rates (and therefore a greater demand for immediate access to minerals) have more osteocytes per bone volume. Finally, osteocytes express molecules typically associated with nerve cells and which are involved with glutamate neurotransmission. By this system, almost instantaneous messages may be transmitted throughout the network, an important feature in cells whose homeostatic function would be utilized on a scale of seconds, rather than hours or days. Experimental procedures for determining the role of the osteocyte in mineral homeostasis would require calcium mobilization from the bone matrix on a relatively immediate time scale. The experimental procedure would then be coupled with a high resolution histomorphometric analysis of lacunar radiographic area and mineral density. Added to this would be an in vitro study of mineral activation capacity via cultured osteocytes treated with PTH. Osteocytic osteolysis would be confirmed by an increase in the demineralized volume of osteocytic lacunae and the identification of a chemical mechanism by which osteocytes can readily access the mineral portion of their immediate bone matrix. It should also be true that a reverse capacity exists by which osteocytes can remineralize their immediate matrix utilizing alkaline phosphatase for example, a chemical which they, like osteoblasts, are known to generate. It is thus proposed that osteocytes are both mechanoreceptors and systemic mineral homeostasis regulators. PMID- 15758445 TI - PTH receptors and apoptosis in osteocytes. AB - Osteocytes comprise a heterogenous population of terminally differentiated osteoblasts that direct bone remodeling in response to applied mechanical loading of bone. Increased osteocyte density accompanies the anabolic effect of PTH in vivo, whereas accelerated osteocyte death may be precipitated by estrogen deficiency or excess glucocorticoid exposure (conditions benefitted by intermittent PTH therapy) and by renal failure (where circulating intact PTH and, especially, PTH carboxylfragments are elevated). Osteocytes express type-1 PTH/ PTHrP receptors (PTH1Rs), which are fully activated by aminoterminal PTH fragments and couple to multiple signal transducers, including adenylyl cyclase and phospholipase C. Activation of PTH1Rs in osteocytes promotes gap junction mediated intercellular coupling, increases expression of MMP-9, potentiates calcium influx via stretch-activated cation channels, amplifies the osteogenic response to mechanical loading in vivo, and regulates apoptosis. Control of osteocyte apoptosis by PTH1Rs is complex, in that intermittent PTH(1-34) administration reduces the fraction of vertebral apoptotic osteocytes at 1 month in adult mice but increases femoral metaphyseal osteocyte apoptosis at 1-2 weeks in young rats. In MLO-Y4 cells, PTH(1-34) prevents apoptosis otherwise induced within 6 hr by dexamethasone. In older studies, large doses of intact PTH(1-84) caused rapid "degenerative" morphologic changes in osteocytes, similar to those described in renal osteodystrophy. We isolated clonal conditionally immortalized osteocytic (OC) cell lines from mice homozygous for targeted ablation of the PTH1R gene. OC cells express abundant (2-3 x 10(6) per cell) receptors specific for the carboxyl(C)-terminus of intact PTH(1-84) ("CPTHRs") but, as expected, do not express PTH1Rs or respond to PTH(1-34). CPTHRs are expressed at much lower levels by other skeletally-derived cell lines. Several highly conserved ligand determinants of CPTHR binding have been identified, including PTH(24-27), PTH(53 54) and the sequence PTH(55-84), loss of which reduces binding affinity by over 100-fold. Human PTH(53-84), like PTH(1-84), PTH(24-84), and PTH(39-84), increases OC cell apoptosis. Ala-scanning mutagenesis to define sequences within PTH(55-84) important for binding and bioactivity is underway. We conclude that osteocytes may be important targets for CPTH fragments that are secreted by the parathyroid glands or generated by peripheral metabolism of intact PTH and that accumulate in blood, especially in renal failure. Studies of functional interplay between responses to CPTHRs and (transfected) PTH1Rs, using receptor-specific ligands in OC cells, should provide new insight into PTH regulation of osteocyte function and survival. PMID- 15758446 TI - The role of calcium channels in osteocyte function. AB - Osteocytic response to stretching, which is potentiated by PTH, is distinct from that of osteoblast to high frequency strain. A MAPK dependent signaling pathway is suggested in the osteoblast response. At least two different types of mechanotransduction pathways are present in bone cells of osteoblastic lineage. PMID- 15758447 TI - Mechanosensation and fluid transport in living bone. AB - The mechanosensory mechanisms in bone include (i) the cell system that is stimulated by external mechanical loading applied to the bone; (ii) the system that transduces that mechanical loading to a communicable signal; and (iii) the systems that transmit that signal to the effector cells for the maintenance of bone homeostasis and for strain adaptation of the bone structure. The effector cells are the osteoblasts and the osteoclasts. These systems and the mechanisms that they employ have not yet been unambiguously identified. The candidate systems will be reviewed. It will be argued that the current theoretical and experimental evidence suggests that osteocytes are the principal mechanosensory cells of bone, that they are activated by shear stress from fluid flowing through the osteocyte canaliculi, and that the electrically coupled three-dimensional network of osteocytes and lining cells is a communications system for the control of bone homeostasis and structural strain adaptation. The movement of bone fluid from the region of the bone vasculature through the canaliculi and the lacunae of the surrounding mineralized tissue accomplishes three important tasks. First, it transports nutrients to the osteocytes in the lacunae buried in the mineralized matrix. Second, it carries away the cell waste. Third, the bone fluid exerts a force on the cell process, a force that is large enough for the cell to sense. This is probably the basic mechanotransduction mechanism in bone, the way in which bone senses the mechanical load to which it is subjected. The mechanisms of bone fluid flow are described with particular emphasis on mechanotransduction. Also described is the cell to cell communication by which higher frequency signals might be transferred, a potential mechanism in bone by which the small whole tissue strain is amplified so the bone cells can respond to it. One of the conclusions is that higher frequency low amplitude strains can maintain bone as effectively as low frequency high amplitude strains. This conclusion leads to a paradigm shift in how to treat osteoporosis and how to cope with microgravity. PMID- 15758448 TI - Microstructural strain near osteocyte lacuna in cortical bone in vitro. AB - Mechanical factors affect bone remodeling such that increased mechanical demand results in net bone formation, whereas decreased demand results in net bone resorption. Two proposed mechanical signals are stress-generated fluid flow forces acting on cells and bone matrix deformation itself. A prominent current theory is that bone cells are more responsive to fluid flow than to mechanical strain. Recent experiments support this conclusion: bone cells increase their production of osteopontin (OPN) mRNA, prostaglandin (PGE(2)), and nitric oxide (NO) in response to fluid flow in contrast to cells stimulated by mechanical strain levels similar to those measured in vivo. However, when cells are subjected to substrate strains levels many times greater than those measured in vivo, increased biological activity again results. We assert that it is neither fluid flow nor matrix deformation per se, but rather the resulting cell deformation that causes cell biological response. Machined specimens of undamaged bovine cortical bone were subjected to increasing levels of macroscopic strain while observed under an optical microscope at 220X. Continuum level strain was measured using a standard foil strain gauge attached to the back of the specimen and ranged from 500 to 6,000 microstrain. Images of the specimen surface at each strain level were captured. To determine the level of osteocyte deformation that results from fluid flow in vitro, MLO-Y4 cells were cultured on collagen coated 190 cm2 plastic sheets and subjected to steady fluid flow at 16 dynes/cm(2). Images representing the initial undisturbed cell configuration and the configuration of the cells after ten minutes of fluid flow were acquired from a videotape of the flow experiment. The captured unloaded vs. loaded image pairs were analyzed to determine the local deformation and strain fields using a digital stereoimaging system. When subjected to a nominal continuum strain level approximately equal to that measured in humans in vivo during rigorous activity (2,000 microstrain), the local, osteocyte level strains can be as high as 12,000 to 15,000 microstrain (1.2% to 1.5%). Average osteocyte strains due to fluid flow in vitro increase from 7,972 microstrains after 16 seconds of flow to 22,856 microstrains after 64 seconds of flow. In contrast, maximum strains measured in vivo are approximately 1,800 microstrain in humans and up to 3,000 microstrain in other species. These data may help to explain why bone cells are more sensitive to fluid flow than substrate strain; fluid forces result in cell deformations much higher than those considered to be "physiological". PMID- 15758449 TI - Skeletal adaptation to mechanical stimuli in the absence of formation or resorption of bone. AB - Too often, unique loading environments fail to alter bone mass and morphology, calling to question the validity of Wolff's Law; the skeleton's plasticity to mechanical signals(1). We propose that bone can accommodate new loading environments without the need to form or resorb tissue, and that a critical aspect of bone tissue's ability to adapt to mechanical stimuli is first achieved via the plasticity of the osteocyte. We suggest that the osteocyte is capable of "normalizing" its local mechanical environment by modulating its cytoskeletal architecture, attachment to the matrix, configuration of the periosteocytic space, and communication channels to surrounding cells. We believe that through this local adaptive mechanism the osteocyte can accommodate the majority of changes in the mechanical milieu without altering the tissue architecture. It is only when bone tissue is subject to more severe (albeit rare) increases or decreases in the functional environment, the osteocyte participates in the formation and/or resorption of bone by coordinating site-specific recruitment of osteoblasts and/or osteoclasts. In vivo models of bone adaptation, combined with in situ reverse transcriptase-PCR, semi-quantitative RT-PCR, Northern analysis, immuno-cytochemistry and histomorphometry, can demonstrate how distinct mechanical stimuli influence the osteocyte's cytoskeletal and lacunar architecture, coupling (and uncoupling) of the osteocyte to the matrix and neighboring cells, and the osteocyte's participation in the recruitment and differentiation of osteoblasts and osteoclasts. Thus, the osteocyte controls three strategies to modulate its local and global environment in response to three distinct functional stimuli: 1) exogenous mechanical stimuli which are distinct from normal but sufficient to maintain bone mass, 2) mechanical stimuli which are osteogenic, and 3) disuse. If it is true that the resident cell population is capable of accommodating subtle changes in the functional milieu before modification of tissue morphology is deemed necessary, a novel strategy for the development of prophylaxes for osteopenia, osseointegration and fracture healing may become apparent. PMID- 15758450 TI - Principles of developmental biology. AB - The field of developmental biology has a history that spans the last 500 years. Within the last 10 years, our understanding of developmental mechanisms has grown exponentially by employing modern techniques of genetics and molecular biology, frequently combined with experimental embryology and the use of molecular markers, rather than solely morphology, to identify critical populations of cells and their state of differentiation. Three main principles have emerged. First, mechanisms of development are highly conserved, both among developing rudiments of a variety of organ systems and among diverse organisms. This conservation occurs both at the level of tissue and cellular mechanisms, and at the molecular level. Second, the development of organ rudiments is influenced by surrounding tissues through interactions called inductive interactions. Such interactions are mediated by highly conserved growth factors and signaling systems. Third, development is a life-long process and can be reawakened in events such as wound healing and regeneration, and in certain diseases. Advances in understanding normal development provide hope that diseases in which development runs amuck, such as cancer, may soon be preventable and fully treatable. Supported by NS 18112 and DC 04185 from the NIH. PMID- 15758451 TI - Regenerative biology and medicine. AB - The replacement of damaged tissues and organs with tissue and organ transplants or bionic implants has serious drawbacks. There is now emerging a new approach to tissue and organ replacement, regenerative biology and medicine. Regenerative biology seeks to understand the cellular and molecular differences between regenerating and non-regenerating tissues. Regenerative medicine seeks to apply this understanding to restore tissue structure and function in damaged, non regenerating tissues. Regeneration is accomplished by three mechanisms, each of which uses or produces a different kind of regeneration-competent cell. Compensatory hyperplasia is regeneration by the proliferation of cells which maintain all or most of their differentiated functions (e.g., liver). The urodele amphibians regenerate a variety of tissues by the dedifferentiation of mature cells to produce progenitor cells capable of division. Many tissues contain reserve stem or progenitor cells that are activated by injury to restore the tissue while simultaneously renewing themselves. All regeneration-competent cells have two features in common. First, they are not terminally differentiated and can re-enter the cell cycle in response to signals in the injury environment. Second, their activation is invariably accompanied by the dissolution of the extracellular matrix (ECM) surrounding the cells, suggesting that the ECM is an important regulator of their state of differentiation. Regenerative medicine uses three approaches. First is the transplantation of cells into the damaged area. Second is the construction of bioartificial tissues by seeding cells into a biodegradable scaffold where they produce a normal matrix. Third is the use of a biomaterial scaffold or drug delivery system to stimulate regeneration in vivo from regeneration-competent cells. There is substantial evidence that non regenerating mammalian tissues harbor regeneration-competent cells that are forced into a pathway of scar tissue formation. Regeneration can be induced if the factors leading to scar formation are inhibited and the appropriate signaling environment is supplied. An overview of regenerative mechanisms, approaches of regenerative medicine, research directions, and research issues will be given. PMID- 15758452 TI - Wnt-signaling and skeletogenesis. AB - Members of the Wnt gene family, encoding secreted cystein-rich glycoproteins, have been isolated from a variety of organisms. They serve as important developmental signaling molecules and have been implicated to play crucial roles in such diverse processes as cancer, organogenesis and pattern formation. Experiments by Zakany and Duboule, and Rudnicki and Brown have suggested a role for Wnt molecules in negatively regulating chondrogenesis. However, neither of the two Wnt genes used in these studies is endogenously expressed in chondrogenic regions. We and others have found that in the chick limb at least four members of the Wnt gene family, Wnt-4, Wnt-5a, Wnt-5b, and Wnt-14, are expressed in defined regions of the developing chondrogenic elements. With the exception of Wnt-5b, which is expressed in perichondrial cells and prehypertrophic chondrocytes, the expression of the three other Wnt genes is restricted to the perichondrium surrounding the cartilage element. Viral misexpression studies in the chick suggested that Wnt-4 acts as a positive signal originating from the joint region and when misexpressed accelerates chondrocyte maturation, while Wnt-5a and Wnt-5b both negatively regulate chondrocyte maturation. We have further shown that they utilize different signaling pathways; while Wnt-4 signals through the canonical Wnt-pathway, Wnt-5a and Wnt-5b do not. Interestingly, the delay in chondrocyte maturation due to Wnt-5a misexpression is associated with an up regulation of Wnt 5b expression in the prehypertropic chondrocytes. Concomitantly, Wnt-5b misexpression also delays chondrocyte maturation. However, preliminary studies suggest that the two Wnt genes affect different steps in the maturation process. Wnt signaling, however, is not only regulating chondrogenesis but is also involved in the segmentation process of the appendicular skeleton. Localized misexpression of the fourth Wnt gene, Wnt-14, which is expressed early in the presumptive joint region, induces morphological and molecular changes indicative of an early joint interzone, suggesting that Wnt-14 plays a pivotal role in the induction of the joint interzone. PMID- 15758453 TI - Mechanical effects on skeletal growth. AB - The growth (i.e. increase of external dimensions) of long bones and vertebrae occurs longitudinally by endochondral ossification at the growth plates, and radially by apposition of bone at the periosteum. It is thought that mechanical loading influences the rate of longitudinal growth. The 'Hueter-Volkmann Law' proposes that growth is retarded by increased mechanical compression, and accelerated by reduced loading in comparison with normal values. The present understanding of this mechanism of bone growth modulation comes from a combination of clinical observation (where altered loading and growth is implicated in some skeletal deformities) and animal experiments in which growth plates of growing animals have been loaded. The gross effect of growth modulation has been demonstrated qualitatively and semi-quantitatively. Sustained compression of physiological magnitude inhibits growth by 40% or more. Distraction increases growth rate by a much smaller amount. Experimental studies are underway to determine how data from animal studies can be scaled to other growth plates. Variables include: differing sizes of growth plate, different anatomical locations, different species and variable growth rate at different stages of skeletal maturity. The two major determinants of longitudinal growth are the rate of chondrocytic proliferation and the amount of chondrocytic enlargement (hypertrophy) in the growth direction. It is largely unknown what are the relative changes in these key variables in mechanically modulated growth, and what are the signaling pathways that produce these changes. PMID- 15758454 TI - Neurotransmitter functions in bone remodeling. PMID- 15758455 TI - Glutamatergic regulation of bone remodeling. AB - L-glutamate (Glu) is the predominant neuromediator in the mammalian central nervous system (CNS). Bone is highly innervated and there is growing evidence of a neural control of bone cell metabolism. The recent discovery of Glu-containing nerve fibers in bone and Glu receptors (GluR) and transporters in bone cells suggest that this neuromediator may also act as a signaling molecule in bone and regulate bone cell function. Our previous studies have demonstrated that ionotropic N-Methyl-D-Aspartate (NMDA) GluR are highly expressed by mammalian osteoclasts. NMDA receptors (NMDAR) are heteromers associating the NR1 subunit and one of the four types of NR2 subunits (NR2A to D). We showed that osteoclasts express NR1, NR2B and NR2D subunits, suggesting a molecular diversity of NMDAR in these cells. Electrophysiological studies have confirmed that NMDAR are functional in mature osteoclasts, and features of Glu-induced current recorded in these cells indicate a major NR2D subunit composition. Using an in vitro assay of bone resorption, we showed that several antagonists of NMDAR binding to different sites of the receptor inhibit bone resorption. In particular, the specific NMDAR channel blocker MK801 had no effect on osteoclast attachment to bone and survival while it rapidly decreased the percentage of osteoclasts with actin ring structures that are associated with actively resorbing osteoclasts. NMDAR may thus be involved in adhesion-induced formation of the sealing zone required for bone resorption. NMDAR are also expressed by osteoclast precursors isolated from mouse bone marrow. We recently confirmed the presence of NR1, NR2B and NR2D in these cells and demonstrated their expression at all differentiation stages from osteoclast precursors to mature resorbing osteoclasts. No regulation of these subunits mRNA expression levels was observed throughout the osteoclastic differentiation sequence. Activation of NMDAR may therefore represent a new mechanism for regulating osteoclast formation and activity. While the origin of Glu in bone is still unknown, the possibility of a glutamatergic neurotransmission in this tissue is suggested by the detection of Glu in nerve fibers in close contact to bone cells. Furthermore, we recently demonstrated that sciatic neurectomy in growing rats induces a bone loss associated with a reduction of nerve profiles immunostained for Glu. These results suggest that Glu may be released from glutamatergic nerve profiles present in bone and therefore contribute to the local regulation of bone cell function. PMID- 15758456 TI - Osteoblastic glutamate receptor function regulates bone formation and resorption. AB - Previous studies showed that a variety of bone cells express protein components necessary for neuronal-like glutamatergic signaling and implicated glutamate as having a role in mechanically induced bone remodeling. Initial functional studies concentrated on the role of glutamate signaling in bone resorption and provided compelling evidence to suggest that glutamate signaling through functional NMDA type ionotropic glutamate receptors (iGluRs) is a prerequisite for in vitro osteoclastogenesis. Originally, effects of iGluR antagonists seen in co-cultures were attributed to antagonists acting directly on osteoclast precursors. However, in the light of recent osteoblast studies it now seems likely that the observed effects on osteoclastogenesis are an indirect effect of modulating the function of pre-osteoblast present within these cultures. The presence of iGluRs in osteoblasts suggests a role for them in bone formation and this paper reviews and discusses the emerging data relating to the role of glutamate signaling in osteoblasts. A number of recently published studies have shown that osteoblasts not only express a wide number of 'pre-synaptic' glutamatergic proteins but also possess the ability to both regulate glutamate release and actively recycle extracellular glutamate. The functionality of osteoblastic 'post-synaptic' glutamatergic components has also been shown as both primary and clonal osteoblasts express electrophysiologically active iGluRs, metabotropic type glutamate receptors (mGluRs) along with a variety of glutamate receptor associated signaling proteins. There is, however, little published data regarding the actual role of glutamatergic signaling in osteoblastic bone formation. In vivo and in vitro studies performed provide evidence that glutamatergic signaling is a necessity for normal osteoblast function. In a number of different models of in vitro bone formation, the addition of non-competitive antagonists of iGluRs prevents the formation of mineralized bone, moreover antagonizing some sub-types of iGluR mediates the differentiation of pre-osteoblasts. iGluR antagonists modulate osteoblast function in a manner that correlates with the previously reported data regarding in vitro osteoclastogenesis. Interestingly iGluR mediated glutamate signaling appears to function differently in osteoblasts derived from flat and long bones. This implies the components of osteoblastic glutamatergic signaling may be adapted in vivo possibly to reflect the differential function of osteoblasts in those regions of the skeleton. PMID- 15758457 TI - The role of dopamine and serotonin in regulating bone mass and strength: studies on dopamine and serotonin transporter null mice. AB - Neurotransmitter regulation of bone metabolism has been a subject of increasing interest and investigation. Dopamine (DA) has been reported to have effects on calcium and phosphorus metabolism. The dopamine transporter (DAT) is believed to control the temporal and spatial activity of released DA by rapid uptake of the neurotransmitter into presynaptic terminals. We have evaluated the histologic and biomechanical properties of the skeleton in mice homozygous for deletion of the DA transporter gene (DAT (-/-)) to help delineate the role of DA in bone biology. We have demonstrated that DAT (-/-) mice have reduced bone mass and strength. DAT (-/-) animals have shorter femur length and dry weight, and lower ash calcium content. Cancellous bone volume in the DAT (-/-) proximal tibial metaphysis is significantly decreased with reduced trabecular thickness. DAT (-/-) vertebrae have lower cancellous bone volume as a consequence of increased trabecular spacing and reduced trabecular number, and cortical thickness and bone area in the femoral diaphysis are reduced. The ultimate bending load (femoral strength) for the DAT (-/-) mice is 30% lower than the wild-type mice. Thus, deletion of the DAT gene results in deficiencies in skeletal structure and integrity. Since serotonin (5-HT) plays a role as a regulator of craniofacial morphogenesis, we explored the expression and function of 5-HT receptors and the 5-HT transporter (5-HTT) in bone. Primary cultures of rat osteoblasts (rOB) and a variety of clonal osteoblastic cell lines including ROS 17/2.8, UMR 106-H5 and Py1a show mRNA expression for the 5-HTT, and the 5-HT(1A), 5-HT(1D), 5-HT(2A) and 5-HT(2B) receptors by RT-PCR analysis and immunoblot. A relatively high density of nanomolar affinity 5-HTT binding sites is present in ROS 17/2.8 and UMR 106-H5 cells. The maximal [(3)H]5-HT uptake rate in ROS cells was 110 pmol/10 min/well, with a K(m) value of 1.13 microM. In normal differentiating rOB cultures, 5-HTT functional activity was observed initially at day 25, and activity increased by almost eight-fold at day 31. In mature rOB cultures, the estimated density of [(125)I]RTI-55 binding sites was 600 fmol/mg protein. PMA treatment caused a significant 40% reduction in the maximal uptake rate of [(3)H]5-HT, an effect prevented by pretreatment with staurosporine. 5-HT potentiates the PTH-induced increase in AP-1 activity in UMR 106-H5 cells. In 5-HTT (-/-) animals, cancellous bone volume (BV/TV) in the lumbar vertebrae is reduced, with a trend toward decreased trabecular thickness and trabecular number. These results demonstrate that osteoblastic cells express a functional serotonin system, with mechanisms for responding to and regulating uptake of 5-HT, and disruption of the 5-HTT gene may cause osteopenia. PMID- 15758458 TI - Harold M. Frost. William. F. Neuman awardee 2001. PMID- 15758459 TI - Why should many skeletal scientists and clinicians learn the Utah paradigm of skeletal physiology? AB - Adding later facts and ideas to a universally accepted "1960 paradigm" of skeletal physiology led to the still-evolving "Utah paradigm". The ASBMR's William Neuman award in 2001 to one of the latter paradigm's architects (HMF) suggested that physiologists began to view it as a valid supplement to its predecessor. Nevertheless it diffused poorly among most SSCs (Skeletal Scientists and Clinicians, plus all others who work in any way on skeletal matters), even though success in the quest for knowledge and recognition by many of them could depend on learning that paradigm's insights. Those insights can help to minimize serious errors in some experimental designs and in interpreting some kinds of data. To explain how success in that quest could depend on the Utah paradigm requires explaining the nature of the above errors, some features of both paradigms, some implications of the newer one, and when that quest's success might not require knowing the Utah paradigm. A three-part message distilled from the past for present and future SSCs concludes the article. It took decades to understand such things and find effective ways to explain them, and both matters probably need improvement (to paraphrase Pogo, "We met the enemy and perhaps it was us more than them"). During those decades the author changed from an active SSC hunter-player to a spectator, known to some as a feisty eccentric old dinosaur (FEOD) (Note A). So here a voice from the past would speak to present and future SSCs. PMID- 15758460 TI - Does the anterior cruciate have a modeling threshold? A case for the affirmative. AB - Growing evidence supports a 1972 proposal that dynamic tension strains of a ligament above a threshold range, but below its ultimate strength, would make its cells synthesize more collagen to thicken and strengthen it. If so, when that strengthening reduced later strains to the bottom of that threshold range this "diametric modeling" would stop. A) Such a mechanism must create a "strength safety factor" that would minimize or prevent voluntary activities from rupturing healthy ligaments, so chiefly injuries would rupture them. B) Such a mechanism should also make the usual largest loads on a healthy anterior cruciate ligament (ACL) determine its strength, and would make smaller loads mixed with large ones have little effect on its strength. C) In principle, when an ACL's strains exceeded that threshold range, diametric modeling would turn on, strengthen it, and reduce subsequent strains from the same loads. When its strains remained smaller, this mechanically-controlled modeling would turn off. Normal ACLs do have a strength-safety factor so they could have a diametric modeling threshold too, as we now know bone does. In healthy young adult humans available evidence suggests that threshold's value could lie in the region of 23 Newtons/mmC of the ACL's cross section area. If similar relationships applied to fascia, tendons and other ligaments (I suggest they do), they would form fundamental biomechanical properties of collagenous tissue organs. PMID- 15758461 TI - Calcitonin effects on cartilage and fracture healing. AB - The literature about the effects of systemically administered calcitonin on fracture healing and in the prevention of disuse osteoporosis after fracture are reviewed in this study. Fracture healing is a biological process of great importance for the survival of the injured animal. Endochondral ossification is augmented in the fracture site followed by fast remodeling of the produced woven bone. There is strong evidence of the direct effects of calcitonin on cartilage proliferation as well as the vascularization of the callus. Calcitonin is found to promote the cartilaginous phase of fracture healing. On the other hand, the innervation of callus reveals an extensive distribution of sensory fibers containing a calcitonin gene-related peptide, a neuropeptide with potent vasodilatory actions. From several experimental studies, salmon calcitonin administration has been found to have a beneficial effect on fracture healing. Studies in humans also concur that calcitonin may speed up the time of fracture repair and facilitate early mobilization of the injured limb. Finally, calcitonin prevents post-fracture bone loss due to increased post-injury remodeling and lowers hydroxyproline and calcium excretion of patients who underwent internal fixation of fracture on the hip. PMID- 15758462 TI - Oestrogen action on bone cells. AB - Sex steroids have an important impact on bone physiology. Oestrogen (E) appears to be the most important sex steroid in preventing osteoporosis in women. Despite the overwhelming evidence that oestrogens modulate bone growth and turnover in vivo, oestrogen receptors (ER) were detected only recently. Two forms of ER have been discovered so far, ERalpha and ERbeta. Both have been detected in osteoblasts and osteoclasts as well. A number of growth factors and cytokines appear to modulate bone resorption in vitro and in vivo. Among others, interleukin-1 and -6 and tumor necrosis factor alpha and beta were found to be extremely potent stimulators of bone resorption. Binding of different cytokines to their receptors in osteoblasts result in the release of soluble factors that act directly on osteoclasts to modulate their recruitment or activity. Thus, E, apart from the direct regulation of osteoclasts, which it achieves through its receptors, can inhibit the release of osteoclast stimulatory factors or enhance the release of osteoclast inhibitory factors. In general, E is an inhibitor of bone resorption that decreases both osteoclast numbers and activity. Recently, it has also been shown that it promotes apoptosis. Moreover, it also has anabolic effects on osteoblasts. However, E action on osteoclasts is superior in comparison with that on osteoblasts. Recent data have shown that transforming growth factor beta (TGFbeta) mediates the actions of E in bone. Following the example of raloxifene it may be proved that the role of TGFbeta in the actions of E in bone is central and has not only academic interest. More data are needed to elucidate this issue. Finally, recent data suggest the importance of E for bone maturation and development of peak bone mass in men. It seems likely that both E and androgens are required for the growth and maintenance of the adult male skeleton. PMID- 15758463 TI - Peripheral quantitative computed tomography at the distal radius: cross calibration between two scanners. AB - Peripheral quantitative computed tomography (pQCT) is an important technique to study the interaction between the muscle and bone systems. We have recently established pQCT reference ranges for children, adolescents and young adults using a recent version (XCT 2000) of the Stratec scanners (Stratec Inc., Pforzheim, Germany). However, the previous version of this type of scanner (XCT 900) is still widely used and cross-calibration is needed to use these reference data. Therefore, both distal radii of 19 healthy subjects (age 21 to 59 years; 11 women) were analyzed at the "4% site" using both the XCT 900 and the XCT 2000. Cross-sectional area, total and trabecular bone mineral density (BMD), total bone mineral content (BMC) and polar Strength-Strain Index (SSI) results from the two scanners were compared using linear regression analysis. To achieve scanner calibration we used the intercept and slope of the correlations. The correlation coefficients between the two devices were 0.82 for the cross-sectional area, 0.81 for total BMD, 0.97 for trabecular BMD, 0.99 for total BMC and 0.86 for polar SSI. In conclusion, these data allow for the conversion of XCT 900 results at the distal radius to XCT 2000 values and vice versa. PMID- 15758464 TI - Postmenopausal changes in the distribution of the volumetric BMD of cortical bone. A pQCT study of the human leg. AB - Three different regions of interest (ROIs) were defined in pQCT scans (XCT-3000 machine, Stratec, Germany) taken at the tibial mid-diaphyses of 12 pre-menopausal (pre-MP) and 12 post-menopausal (post-MP) women who were otherwise normal, according to the volumetric bone mineral density (vBMD) value of their corresponding pixels (voxels) as assessed by their respective attenuation values. They were classified as "low-vBMD" (LD-ROI, with a vBMD of 200-400 mg/cm(3)), corresponding chiefly to trabecular-subcortical bone; "medium-vBMD" (MD-ROI, vBMD = 400-800 mg/cm(3)), corresponding mainly to porous cortical bone or cortical subcortical bone, and "high-vBMD" (HD-ROI, vBMD higher than 800 mg/cm(3)), corresponding to dense cortical bone. The fraction of the total cross-sectional bone area covered by the HD-ROI was 16% higher, and that covered by the MD-ROI 20% lower, in pre-MP than post-MP women. No differences concerning the LDROIs were found. A close, linearly negative relationship was found between the MD- and HD-ROI fractions in all the women together, with no inter-group differences in slope. The Stress-Strain Index (an indicator of the torsional stiffness and strength of the whole bones that involved both the vBMD and the spatial disposition of the HD bone in the cross-section - torsional moment of inertia -) correlated linearly and positively with the cross-sectional area of the HD-ROI, with a higher slope for pre-MP than post-MP women. Qualitatively, a. post-MP women showed a significantly more prevalent discontinuity of the voxels in the HD ROI than pre-MP women, and b. the tendency of LD-ROIs to accumulate along the mechanically lesseffective (antero-posterior) axis of the image - a characteristic of pre-MP bones - was visually less evident in post-MP bones. These features describe non-invasively some changes induced by menopause in the human tibia that may be critical for defining the skeletal condition and to monitor the effects of treatments addressed either to protect or to improve mechanically the bone structure, beyond the possibilities of standard densitometry. PMID- 15758465 TI - Longitudinal measurement of bone mineral density at the radius in hemodialysis patients using dual-energy X-ray absorptiometry. AB - The object of our study was to document the changes in bone mineral density (BMD) at the (1/3) distal radius in patients undergoing maintenance hemodialysis (HD). Forty nine male and 24 female patients were enrolled in this study. The mean age was 55.9-/+13.1 (mean -/+ SD) years, and the duration of HD was 89.2 -/+ 81.0 months at the beginning of the investigation. BMD was measured by dual-energy X ray absorptiometry at 1-year intervals for a period in excess of 3 years. No significant relationship was observed between BMD and age in both sexes. In male patients, BMD was positively correlated with body mass index (BMI) (r=0.47, p<0.01) and negatively with the duration of HD (r=0.61, p<0.01). In contrast, BMD was not correlated with either BMI or with the duration of HD in female patients. Eleven of the 14 patients on HD for more than 15 years showed marked bone loss (male; 0.460, female; 0.394g/cm(2)), although they were relatively young (mean age: 43.4 years). Prolonged HD could be one of the risk factors responsible for bone loss. PMID- 15758466 TI - Interaction between nandrolone decanoate and calcitonin in bone formation markers (osteocalcin and bone specific alkaline phosphatase) and IGF-I in rats. AB - Bone tissue has been shown to contain numerous cell-to-cell signaling peptides called growth factors. These growth factors are thought to have important regulating effects for bone remodeling, due to their potent effects on bone cell metabolism. Our investigation was intended to assess the effect of nandrolone decanoate and calcitonin treatment on biochemical markers of bone formation (bone alkaline phosphatase - osteocalcin) and insulin-like growth factor-I in rats. We studied 48 adult male rats. The animals were divided into four groups. Group (A) served as control. Animals in Group (B) were injected with 4 mg/kg/day nandrolone decanoate. Animals in Group (C) were injected with 400mU/rat/day calcitonin and Group (D) received combined therapy for seven days. Nandrolone decanoate and calcitonin have a mild but significant effect on insulin-like growth factor-I without affecting osteocalcin levels, while calcitonin alone decreases the BALP levels. The coadministration of two agents caused notable elevation on insulin like growth factor-I, followed by a significant increase of osteocalcin and bone alkaline phosphatase. PMID- 15758468 TI - What is new in neuro-musculoskeletal interactions? PMID- 15758467 TI - Localization of cathepsins G and L in spontaneous resorption of intervertebral discs in a rat experimental model. AB - To determine the involvement of cathepsins G and L in the mechanism of spontaneous resorption of herniated intervertebral discs, localization of these cathepsins in this process was examined immunohistochemically using a rat model of autologous transplantation of coccygeal discs. Rat coccygeal discs were resected and autotransplanted into the subcutaneous space of the skin of the back. Paraffin-embedded sections of intervertebral disc tissue, harvested at various post-transplantational periods, were immunohistochemically stained with antibodies for cathepsin G, cathepsin L, MMP-1, MMP-3 and ED-2. The number of positive cells was counted in each part of the transplanted discs. Immunolocalization of cathepsins G and L in various types of disc cells was first observed early in the post-transplantation period. From two days after the operation, histology showed invasion by granulation tissue, with many macrophages, in all sections. Subsequently, the number of macrophages in granulation tissue was observed to increase, along with a gradual increase in the percentage of cells positive for MMP-1 and MMP-3. In addition to the ability of cathepsins G and L to degrade major extracellular matrix components of intervertebral discs, cathepsin G is capable of activating latent pro-MMPs. The up-regulation of cathepsins G and L in the intervertebral disc tissue in this spontaneous resorption model suggests that these proteinases may be involved in degradation of extracellular matrix, leading to the natural resorption of herniated discs. PMID- 15758470 TI - Bone anabolic agents. Introduction. PMID- 15758471 TI - An integrated approach to assess structure-to-function relationships in the skeleton. AB - In general, the disciplines of biomechanics, morphology, densitometry, biochemistry, cell biology and molecular biology have advanced independently of one another. In spite of this fragmentation, there have been incremental increases in our understanding of the organization, mechanical properties, growth, remodeling and repair of the tissues comprising the skeleton. As a practical application, this increased knowledge has greatly improved our capabilities for early diagnosis of bone loss and has proven similarly useful in determining the efficacy of interventions to prevent osteoporosis. This approach, however, has been much less successful in countering several other important musculoskeletal disorders, including arthritis. In the immediate future, a major emphasis will be placed on tissue regeneration (engineering) to restore lost mechanical function to a compromised skeleton. To accomplish this goal, it will be necessary to employ much more sophisticated approaches toward evaluating the structure-to-function relationships, ones which will include integration of the respective contributions of gene expression, cell number and activity, matrix composition and architecture to achieve adequate tissue function. PMID- 15758472 TI - Skeletal effects of systemic treatment with basic fibroblast growth factor. AB - Systemic treatment of intact and ovariectomized rats with basic fibroblast growth factor (bFGF) has strong bone anabolic effects. These effects include marked increases in osteoblast number and activity along cancellous and endocortical bone surfaces, which result in accumulation of osteoid and augmentation of cancellous and cortical bone mass after only short-term treatment with bFGF. Osteoclast surface is markedly decreased in bFGF-treated rats, but this finding may be secondary to the extensive osteoid surface in these animals. Some undesirable skeletal effects of the growth factor include impaired bone mineralization and formation of structurally inferior woven bone. The lack of a bone anabolic response to bFGF at skeletal sites with fatty marrow and along the periosteal surface of cortical bone is also disappointing. Despite these disadvantages, bFGF stimulates cancellous bone formation to such a great extent that it may eventually be considered for use in patients with severe osteoporosis who are unresponsive to conventional therapies, provided that local delivery of the growth factor to bone can be achieved to avoid systemic side effects. PMID- 15758473 TI - Effects of amylin and adrenomedullin on the skeleton. AB - Amylin and adrenomedullin are related peptides with some homology to both calcitonin and calcitonin gene-related peptide (CGRP). All these peptides have in common a 6-amino acid ring structure at the amino-terminus created by a disulfide bond. In addition, the carboxy-termini are amidated. Both amylin and adrenomedullin have recently been found to stimulate the proliferation of osteoblasts in vitro, and to increase indices of bone formation in vivo when administered either locally or systemically. Both amylin and adrenomedullin have also been found to act on chondrocytes (Cornish et al., submitted for publication), stimulating their proliferation in culture and increasing tibial growth plate thickness when administered systemically to adult mice. Studies of structure-activity relationships have demonstrated that osteotropic effects of amylin and adrenomedullin can be retained in peptide fragments of the molecules. The full-length peptide of amylin has known effects on fuel metabolism, and systemic administration of amylin is also associated with increased fat mass. However, the octapeptide fragment of the molecule, amylin-(1-8), is osteotropic and yet has no activity on fuel metabolism. Similar fragments of adrenomedullin have also been defined, which retain activity on bone but lack the parent peptide's vasodilator properties. Both amylin-(1-8) and adrenomedullin-(27-52) act as anabolic agents on bone, increasing bone strength when administered systemically. Thus, these small peptides, or analogues of it, are potential candidates as anabolic therapies for osteoporosis. Both amylin and adrenomedullin may have effects on bone metabolism. Amylin is secreted following eating and may direct calcium and protein absorbed from the meal into new bone synthesis. Amylin circulates in high concentrations in obese individuals, and might contribute to the association between bone mass and fat mass. Our recent findings demonstrating the co-expression of adrenomedullin and adrenomedullin receptors in osteoblasts, along with the findings that the peptide and its receptor are easily detectable during rodent embryogenesis, suggest that this peptide is a local regulator of bone growth. Thus, the findings reviewed in this paper illustrate that amylin and adrenomedullin may be relevant to the normal regulation of bone mass and to the design of agents for the treatment of osteoporosis. PMID- 15758474 TI - Bone anabolic therapy with selective prostaglandin analogs. AB - Prostaglandin E(2) has been shown to increase bone mass in animals and humans but it also has considerable dose limiting systemic side effects. The molecular description of multiple seven transmembrane domain G protein coupled prostanoid receptors offered the opportunity to probe the skeletal effects of specific receptors using selective agonists. Bone effects have been reported with many of the prostanoid receptors, with most interest focused on the anabolic effects of EP2, EP4, and FP receptors. Current data suggests activity at the EP2 receptor stimulates formation, activity at the EP4 receptor stimulates resorption (and possibly formation), and activity at the FP receptor produces new trabeculae. However, caution must be exercised in extending the effects of prostanoids in isolated systems to systemic skeletal effects, since tissue level effects are the cumulative result of bone formation and bone resorption. Furthermore, species differences in receptor sequence and density confound extrapolation of effects from one model to another model. While these molecular targets increase our insight into how the skeleton can be affected pharmacologically, they still do not answer questions about the role of naturally occurring prostaglandins in skeletal health. This manuscript will review some of the recent advances in knowledge of the bone anabolic effects of selective prostanoid ligands. PMID- 15758475 TI - Anabolic actions of PTH in the skeletons of animals. AB - A brief historical perspective reviews studies that tested the hypotheses that PTH induces an anabolic effect in bone, and that the gain in trabecular bone was not at the expense of cortical bone. As PTH reduces the risk of fracture in humans with osteoporosis, the myths that postulated cortical bone porosity and increased bone turnover might increase fracture risk, are examined in the light of data from animals with osteonal bone. These show that PTH "braces" the bone by immediately stimulating bone formation at modeling and remodeling sites. Increased porosity is a late event, occurring close to the neutral axis of bone where detrimental effects on biomechanical strength are unlikely. PTH increases bone mass by stimulating modeling in favor of bone formation, and restructures bone geometry via more extensive remodeling. Cell and genetic events induced in bone by PTH have been studied in rats and are time- and regimen-dependent. In addition to the stimulation of gene expression for matrix proteins, early genes upregulated by once daily PTH are those associated with matrix degradation and induction of osteoclastic resorption, indicative of possible mechanisms by which PTH may increase bone turnover. Boneforming surfaces are increased due to increased numbers of newly differentiated osteoblasts and retention of older osteoblasts by inhibition of apoptosis. After stopping treatment, the number of osteoblasts is quickly reduced and bone turnover returns to that of controls, slowing both bone formation and resorption. The increased proportion of bone undergoing PTH-induced remodeling requires maturation and completion of mineralization. These responses may explain the delay in reversal of gains in bone mass and biomechanical properties for at least two turnover cycles following withdrawal in large animal models. Thus, the skeletal benefits of PTH extend beyond the active treatment phase. PMID- 15758476 TI - The effects of growth hormone on cortical and cancellous bone. AB - Growth hormone (GH) has profound effects on linear bone growth, bone metabolism and bone mass. The GH receptor is found on the cell surface of osteoblasts and osteoclasts, but not on mature osteocytes. In vitro, GH stimulates proliferation, differentiation and extracellular matrix production in osteoblast-like cell lines. GH also stimulates recruitment and bone resorption activity in osteoclast like cells. GH promotes autocrine/paracrine insulin-like growth factor 1 (IGF-I) production and endocrine (liver-derived) IGF-I production. Some of the GH-induced effects on bone cells can be blocked by IGF-I antibodies, while others cannot. In animal experiments, GH administration increases bone formation and resorption, and enhances cortical bone mass and mechanical strength. When GH induces linear growth, increased cancellous bone volume is seen, but an unaffected cancellous bone volume is found in the absence of linear growth. Patients with acromegaly have increased bone formation and resorption markers. Bone mass results are conflicting because many acromegalics have hypogonadism, but in acromegalics without hypogonadism, increased bone mineral density (BMD) is seen in predominantly cortical bone, and normal BMD in predominantly cancellous bone. Adult patients with growth hormone deficiency have decreased bone mineral content and BMD. GH therapy rapidly increases bone formation and resorption markers. During the first 6-12 months of therapy, declined or unchanged BMD is found in the femoral neck and lumbar spine. All GH trials with a duration of two years or more show enhanced femoral neck and lumbar spine BMD. In osteoporotic patients, GH treatment quickly increases markers for bone formation and resorption. During the first year of treatment, unchanged or decreased BMD values are found, whereas longer treatment periods report enhanced or unchanged BMD values. However, existing trials comprising relatively few patients and limited treatment periods do not allow final conclusions to be drawn regarding the effects of GH on osteoporosis during long-term treatment. PMID- 15758477 TI - Vitamin D analogs and bone. AB - Vitamin D analogs increase intestinal calcium absorption, and have been shown to possess antiresorptive and also bone anabolic properties in vivo. Therefore, the pharmacological profile of vitamin D analogs would be well suited for the treatment of osteoporosis. However, the calcemic side effects of this compound class, especially at higher doses, have hampered their wide use in osteoporotic patients. Nevertheless, the clear potential for bone anabolic properties together with oral availability have stimulated the interest in this substance class, and there is an active search for bone selective vitamin D compounds. After an overview of the physiological functions of vitamin D in bone, this article focuses on the effects of acute and chronic administration of pharmacological doses of vitamin D analogs on bone in animal models and humans. Furthermore, the endocrinological, cellular, and microanatomical mechanisms involved in the skeletal actions of vitamin D analogs are discussed. The final section briefly reviews the available data on possible bone selective vitamin D analogs. PMID- 15758478 TI - Multi-lineage potential of human mesenchymal stem cells following clonal expansion. AB - Bone marrow contains mesenchymal cells that can be isolated and grown in vitro. Using appropriate treatment protocols such cultures can be induced to differentiate to yield osteoblasts, adipocytes, and chondrocytes. However, previous experiments had not addressed the question whether single pluripotent stem cells exist and can give rise to these different cell lineages or whether bone marrow mesenchymal cell preparations represent a mixture of committed precursors. We have used human adult bone marrow-derived mesenchymal cells obtained from iliac crest biopsies to demonstrate clonal outgrowth after limiting dilution and we show that some clones can be expanded over more than 20 cumulative population doublings and differentiated to osteoblasts, adipocytes, and chondrocytes. Our data provide direct experimental evidence that cultures of bone marrow-derived mesenchymal cells contain individual cells that fulfil two essential stem cell criteria: (i) extensive self-renewal capacity and (ii) multi lineage potential. PMID- 15758479 TI - A comparison of the anabolic effects of rat and bovine parathyroid hormone (1-34) in ovariectomized rats. AB - The current study was designed to compare the skeletal effects of comparable doses of rat parathyroid hormone 1-34 (rPTH) and bovine parathyroid hormone 1-34 (bPTH) in ovariectomized (OVX) rats. Female Sprague-Dawley rats were OVX or sham operated at 6 months of age and maintained untreated for 28 days after surgery. Baseline control and OVX rats were sacrificed at the beginning of treatment. Beginning 28 days post-OVX, the remaining rats were subcutaneously injected daily with rPTH or bPTH at 0, 5, 25, or 50 microg/kg/d for 28 days. Bone area, bone mineral content (BMC), and bone mineral density (BMD) of the distal femoral metaphyses were determined ex vivo using dual energy X-ray absorptiometry. Quantitative bone histomorphometry was performed on undecalcified longitudinal sections of the proximal tibia from each rat. Baseline OVX rats exhibited osteopenia as demonstrated by their significantly reduced femoral BMD and proximal tibia cancellous bone volume compared with those of baseline sham controls. Both rPTH and bPTH restored bone in OVX rats by markedly stimulating bone formation in a dose-dependent manner. However, a difference in potency between the two forms of PTH was evident. The percentage increases of BMC, BMD, cancellous bone volume, trabecular thickness, mineralizing surface, and bone formation rate in the OVX rats treated with bPTH at 5 microg/kg/d were the same as or above those treated with rPTH at the 25 microg/kg/d dose level. A relative potency analysis showed that bPTH was approximately 4- to 6-fold relatively more potent than rPTH in increasing distal femoral BMD as well as cancellous bone volume, mineralizing surface, and bone formation rate of proximal tibial metaphyses at comparable dose levels and a given time. These results may serve as a reference for in vivo study design when rPTH or bPTH are to be the agents for studies on bone anabolism. PMID- 15758481 TI - Calcitonin, an enigmatic hormone: does it have a function? AB - This editorial presents our view of the status of thyroidal calcitonin (TCT) in mammalian physiology. The discovery of calcitonin (CT) enabled the development of a valuable therapeutic agent but the early experiments most likely misled us with regard to its physiological significance. These early purported roles for TCT, first as an agent important in blood calcium regulation and later as an agent to prevent hypercalcemia, are no longer considered as physiological functions. While large supraphysiological doses of CT have an effect on the morphology and function of osteoclasts, it is unlikely that these effects of CT are important in the normal physiology of osteoclasts or bone remodeling. It is surprising that 38 years after the discovery of TCT there is no consensus as to its role in normal mammalian physiology. This editorial concerns three possibilities with respect to TCT: 1) the hormone is vestigial; 2) the hormone plays a role in water metabolism, ionic concentrations, and/or acid-base balance, actions that may not involve calcium metabolism at all; and 3) TCT acts to store phosphate postprandially on bone surfaces as a labile calcium-phosphate colloid, an action that may provide calcium needed for use in non-feeding periods or to reduce postprandial loss of phosphate when dietary phosphate is limited. Also discussed are recent publications indicating that CT synthesized in non-thyroidal tissues (NTCT) may have paracrine actions. PMID- 15758482 TI - Perspectives on using nonhuman primates to understand the etiology and treatment of postmenopausal osteoporosis. AB - The reproductive physiology and skeletal anatomy of nonhuman primates are very similar to those of women and these similarities have prompted studies of the effects of ovariectomy in monkeys on bone metabolism. Following ovariectomy, monkey bone exhibits increases in remodeling activity resulting in bone loss. Since similar bone changes occur after menopause in women, ovariectomized monkeys provide an excellent model of the early skeletal events following menopause and have been employed to study the skeletal actions of drugs designed to treat postmenopausal osteoporosis. This review describes the motivations for examining monkeys, practical aspects of working with monkeys, comparisons of human and monkey bone anatomy, endocrinological aspects of monkey bone metabolism, and the available data obtained in monkeys related to postmenopausal and other forms of osteoporosis. PMID- 15758483 TI - Osteocyte-osteoclast morphological relationships and the putative role of osteocytes in bone remodeling. AB - An osteocyte lacunae differential count under the light microscope (LM) (1 lacunae with live osteocytes, 2-empty lacunae and lacunae with degenerating osteocytes) was carried out outside the reversal lines of osteonic lamellar bone from various mammals and man to evaluate the possibility of osteocyte survival where osteoclast resorption had occurred. The polarized light microscope (PLM) was used to establish the curvature of bony lamellae outside the convexity of reversal lines: concave lamellae indicate osteocytes reabsorbed on their vascular side where they radiate long vascular dendrites; convex lamellae indicate bone resorption on the osteocyte mineral side, radiating short dendrites. In all samples it was found that: a) about 60% of osteocytes outside the reversal lines were live; b) the percentage of alive osteocytes close to reversal lines is higher when they are attacked on their mineral side. The present data support our view that surviving osteocytes, particularly those attacked from their mineral side, might intervene in the final phase of bone resorption (osteoclast inhibition?). The fact that under the transmission electron microscope (TEM) intercellular contacts were never observed between osteocytes and osteoclasts indicates that if a modulation should occur between these two cellular types it could take place by a paracrine route only. The putative role of the cells of the osteogenic system, particularly osteocytes, in the bone remodeling cycle is also discussed. PMID- 15758484 TI - Animal models relevant to cementless joint replacement. AB - This review focuses on animal models used to study certain aspects of 'cementless' joint replacement. Implants used in this application are designed to become attached to the host skeleton through either bone ingrowth into porous surfaces or bone apposition (ongrowth) onto other types of surfaces. Biological fixation of cementless joint replacement implants relies on intramembranous bone regeneration. We describe a framework for understanding research design in light of the type of research questions now being asked. In particular, species choice, implant design and placement, and experimental endpoints are described in some detail. We provide a summary of recent studies specifically focused on implant fixation, demonstrating that most work is still at the morphological and biomechanical levels with little understanding at the molecular level. We also provide a more comprehensive listing of studies using hip and knee replacement models, demonstrating that most work is focused on the interface, and responses of the immediately adjacent trabecular bone and the more distant cortical bone. We conclude by encouraging investigators to design their experiments so that there is enough power to answer a limited number of questions as opposed to providing limited data on a broader number of issues. PMID- 15758485 TI - Skeletal adaptations during mammalian reproduction. AB - Remarkable changes occur in the mammalian skeleton prior to, during and after the reproductive cycle. Skeletal changes occur with ovarian maturation and initiation of menses and estrus in adolescence, which may result in a greater accumulation of skeletal mineral in the female vs the male skeleton. There is also some evidence to suggest an excess skeletal mass in young female experimental animals. In early pregnancy, growth, modeling and perhaps suppressed remodeling promote the accumulation of calcium. Some changes may also occur with the transition from pituitary to placental control of the pregnancy. In later pregnancy, an increase in bone turnover appears to coincide with fetal skeletal mineralization. Rapid and important changes occur in the skeleton and mineral metabolism in the transition from pregnancy to lactation as the mammary gland rather than the uterus draws on the maternal calcium stores. Lactational demands are met at least partially by a temporary demineralization of the skeleton, which is associated with increased bone modeling and remodeling. Endochondral growth almost ceases during lactation, but envelope-specific bone modeling and remodeling are greatly increased. This is generally associated with a loss of skeletal mass and density, more apparent at sites with less of a mechanical role (e.g. central metaphysis regions and the endocortical envelope). The post-lactational period is profoundly anabolic with substantial increases in bone formation, but blunted resorption at almost all skeletal envelopes. Skeletal mass is increased during this period and it is associated with improved skeletal mechanical properties. There are several important observations. 1) The nulliparous animal appears to have an excess skeletal mass to perhaps compensate for maternal metabolic inefficiency of the first reproductive cycle. 2) Changes in growth, modeling and remodeling occur at different times and at different skeletal envelopes during the reproductive cycle. These site-specific, temporal changes appear to be adaptations that facilitate the use of skeletal mineral while preserving mechanical competence. 3) After the first reproductive cycle, modeling and remodeling optimize the existing skeletal mass into a structure that better accommodates the prevailing mechanical environment. 4) The post-lactational period is profoundly anabolic and may provide new strategies for preservation of skeletal mass when reproductive capacity ceases. PMID- 15758486 TI - Overload arthrosis: strain patterns in the equine metacarpal condyle. AB - An overload arthrosis occurs consistently in the palmar region of the metacarpal condyle of the equine fetlock (metacarpophalangeal) joint characterized by subchondral bone sclerosis, devitalization and mechanical failure leading to collapse of the overlying articular cartilage. Samples were selected of joints with mild, moderate, and severe subchondral sclerosis, in which cartilage collapse had not yet occurred. An additional group that had severe sclerosis with focal rarefaction suggesting impending collapse was also studied (n=5/group). Parasagittal slices were milled to 2.0 mm thickness and subjected to palmar forces 50 to 200% of those applied by the sesamoid bone at angles corresponding to early, mid and late stance support phases of the gait cycle. From contact radiographs in the loaded and unloaded samples, strains were determined by recognizing displacements in the trabecular patterns using texture correlation analysis. Failure did not occur in any of the samples. Strains were generally proportional to the forces applied and greatest at midstance. Strain patterns varied between samples and with the different loading positions. With increased subchondral bone sclerosis there was greater shear strain in overlying trabeculae. Strain patterns were not consistently different within the sclerotic bone at the site of failure. Focally higher strains at the surface were sometimes related to the edge of the platen which was molded to mimic the sesamoid bone in vivo. These results indicate that sclerotic thickening of subchondral bone transmits stresses to overlying trabeculae. No consistent strain pattern was recognized where devitalization and mechanical failure occurs. Focally higher strains related to the edge of the opposing sesamoid bone may play a role. PMID- 15758487 TI - Animal models of osteoarthritis. AB - Animal models of osteoarthritis are used to study the pathogenesis of cartilage degeneration and to evaluate potential antiarthritic drugs for clinical use. Animal models of naturally occurring osteoarthritis (OA) occur in knee joints of guinea pigs, mice and other laboratory animal species. Transgenic models have been developed in mice. Commonly utilized surgical instability models include medial meniscal tear in guinea pigs and rats, medial or lateral partial meniscectomy in rabbits, medial partial or total meniscectomy or anterior cruciate transection in dogs. Additional models of cartilage degeneration can be induced by intra-articular iodoacetate injection or by administration of oral or parenteral quinolone antibiotics. None of these models have a proven track record of predicting efficacy in human disease since there are no agents that have been proven to provide anything other than symptomatic relief in human OA. However, agents that are active in these models are currently in clinical trials. Methodologies, gross and histopathologic features and comparisons to human disease will be discussed for the various models. PMID- 15758488 TI - Animal models of rheumatoid arthritis. AB - Animal models of arthritis are used to study pathogenesis of disease and to evaluate potential anti-arthritic drugs for clinical use. Therefore morphological similarities to human disease and capacity of the model to predict efficacy in humans are important criteria in model selection. Animal models of rheumatoid arthritis (RA) with a proven track record of predictability for efficacy in humans include: rat adjuvant arthritis, rat type II collagen arthritis, mouse type II collagen arthritis and antigen-induced arthritis in several species. Agents currently in clinical use (or trials) that are active in these models include: corticosteroids, methotrexate, nonsteroidal anti-inflammatory drugs, cyclosporin A, leflunomide interleukin-1 receptor antagonist (IL-1ra) and soluble TNF receptors. For some of these agents, the models also predict that toxicities seen at higher doses for prolonged dosing periods would preclude dosing in humans at levels that might provide disease modifying effects. Data, conduct and features of the various models of these commonly utilized models of RA as well as some transgenic mouse models and less commonly utilized rodent models will be discussed with emphasis on their similarities to human disease. PMID- 15758489 TI - Changes in trabecular bone turnover and bone marrow cell development in tail suspended mice. AB - Skeletal unloading induces trabecular bone loss in loaded bones. The tail suspended mouse model simulates conditions associated with lack of mechanical stress such as space flight for the loaded bones. In such a model, the tail supports the body weight. The forelimbs are normally loaded and the movement of its hindlimbs is free without weight bearing. Histomorphometric analyses of the murine tibiae of the elevated hindlimbs show that trabecular bone volume rapidly diminishes within one week and stabilizes at that level in the subsequent week of tail suspension. Two-week reloading after one-week unloading completely restores trabecular bone volume, but this does not happen after two-week unloading. Unloading for one or two weeks significantly reduces bone formation rate and increases both the osteoclast surface and number compared with age-matched ground control mice. Subsequent reloading restores reduced bone formation and suppresses increased bone resorption. In bone marrow cell cultures, the numbers of alkaline phosphatase (ALP)-positive colony-forming units-fibroblastic (CFU-f) and mineralized nodules are significantly reduced, but the numbers of adherent marrow cells and total CFU-f are unaltered after tail suspension. On the other hand, subsequent reloading increases the number of adherent marrow cells. Unloading for one week significantly increases the number of tartrate-resistant acid phosphatase (TRAP)- positive multinucleated cells compared with the control level. Our data demonstrate that tail suspension in mice reduces trabecular bone formation, enhances bone resorption, and is closely associated with the formation of mineralized nodules and TRAP-positive multinucleated cells in bone marrow cultures obtained from tibiae. Two-week reloading restores bone volume reduced after one-week unloading, but does not after two-week unloading. The tail suspended model provides a unique opportunity to evaluate the physiological and cellular mechanisms of the skeletal response to unloading and reloading. PMID- 15758490 TI - What is new in neuro-musculoskeletal interactions? PMID- 15758491 TI - Animal models of bone diseases. Introduction. PMID- 15758492 TI - The Utah paradigm on animal models of skeletal disorders: quo vadis? AB - Skeletal disorders that need effective studies in suitable animal models include "osteoporosis", arthroses and hard and soft tissue healing. For people doing or analyzing such studies this article provides a brief overview and some salient implications of the Utah paradigm of skeletal physiology. The article leaves discussing and resolving any disagreements and controversies about such things to other times, places and people. PMID- 15758493 TI - Overview: animal models of osteopenia and osteoporosis. AB - Prior to initiating a clinical trial in a post-menopausal osteoporosis study, it is reasonable to recommence the evaluation of treatment in the 9-month-old ovariectomized female rat. A female rat of this age has reached peak bone mass and can be manipulated to simulate clinical findings of post-menopausal osteoporosis. Ample time exists for experimental protocols that either prevent estrogen depletion osteopenia or restore bone loss after estrogen depletion. More time can be saved by acceleration of the development of the osteopenia by combining ovariectomized (OVX) plus immobilization (IM) models. Methods like serum biochemistry, histomorphometry and densitometry used in humans are applicable in rats. Like most animal models of osteopenia, the rat develops no fragility fractures, but mechanical testing of rat bones substitutes as a predictor of bone fragility. Recent studies have shown that the prevailing activity in cancellous and cortical bone of the sampling sites in rats is remodeling. The problems of dealing with a growing skeleton, the site specificity of the OVX and IM models, the lack of trabecular and Haversian remodeling and the slow developing cortical bone loss have been and can be overcome by adding beginning and pre-treatment controls and muscle mass measurements in all experimental designs, selecting cancellous bone sampling sites that are remodeling, concentrating the analysis of cortical bone loss to the peri medullary bone and combining OVX and IM in a model to accelerate the development of both cancellous and cortical bone osteopenia. Not to be forgotten is the distal tibia site, an adult bone site with growth plate closure at 3 months and low trabecular bone turnover and architecture similar to human spongiosa. This site would be most challenging to the action of bone anabolic agents. Data about estrogen-deplete mice are encouraging, but the ovariectomized rat model suggests that developing an ovariectomized mouse model as an alternative is not urgent. Nevertheless, the mouse model has a place in drug development and skeletal research. In dealing with drug development, it could be a useful model because it is a much smaller animal requiring fewer drugs for screening. In skeletal research mice are useful in revealing genetic markers for peak bone mass and gene manipulations that affect bone mass, structure and strength. When the exciting mouse glucocorticoid-induced bone loss model of Weinstein and Manolagas is confirmed by others, it could be a significant breakthrough for that area of research. Lastly, we find that the information generated from skeletal studies of nonhuman primates has been most disappointing and recommend that these expensive skeletal studies be curtailed unless it is required by a regulatory agency for safety studies. PMID- 15758494 TI - Lean, fat and bone masses are influenced by orchidectomy in the rat. A densitometric X-ray absorptiometric study. AB - In man, hypogonadism is a risk factor for osteoporosis. Orchidectomy (ORX) in the rat leads to an imbalance between resorption and formation resulting in bone loss. We have measured whole body weight, lean and fat mass, whole bone mass (BMC) in the ORX rat model by dual X-ray densitometry (DXA). Forty-eight male Wistar rats (18-19 weeks old) were studied at 2, 4, 8 and 16 weeks. In each group, 6 rats were ORX and 6 sham-operated were used as control. DXA was performed on the whole body and isolated tibia. The whole body weight of the ORX animals became significantly decreased only at 16 weeks. Whole body BMC was reduced from 8 weeks in the ORX group. The most striking result was a net decrease in lean mass that reached -15.7% at 16 weeks. On the other hand, fat mass remained unchanged during the time series in the ORX animals. PMID- 15758495 TI - Long-term effects of aging and orchidectomy on bone and body composition in rapidly growing male rats. AB - The purpose of this study was to assess the long-term effects of aging and sex hormone deficiency on skeletal metabolism and body composition in rapidly growing male rats. Sprague-Dawley male rats were sham-operated (sham) or orchidectomized (ORX) at 3 months of age. Eight sham rats and eight ORX rats at each time point were serially sacrificed at 3, 4, 8, 12, 15, and 23 months of age. Bone mass in sham rats rapidly increased until 8 months of age, then slightly increased between 8 to 12 months of age; thereafter, an age-related decrease in bone mass was found between 12 to 23 months of age. In sham rats, bone formation parameters decreased between 3 and 8 months, and maintained at the lower level between 8 and 23 months of age, while bone resorption parameters decreased between 3 and 12 months, and thereafter, increased with age between 12 and 23 months of age. ORX significantly inhibited age-related gain in body weight, lean body mass, and cancellous and cortical bone mass and decreased peak bone mass (approximately 20% less versus sham). Further, we found that the lower bone and lean body mass in ORX rats was due to the lack of age-related gain rather than the net loss from basal controls. These data suggest that sex hormones are important factors for the accumulation of peak bone and lean body mass in male rats. PMID- 15758496 TI - Skeletal effects of androgen withdrawal. AB - Hypogonadism is considered to be one of the major risk factors for osteoporosis in men. Therefore, it is an important goal for skeletal research to improve our understanding of the skeletal effects of androgens. Androgen deficiency during growth is associated with a failure to acquire normal peak bone mass, and there is good evidence that the effects of androgens on skeletal growth and the development of a male skeletal phenotype are mediated through the androgen receptor. In adult men, acute withdrawal of androgens by surgical or chemical castration induces high turnover bone loss. Similarly, orchidectomy of aged, non growing male rats is associated with a pronounced and sustained increase in bone turnover and with true loss of cancellous and cortical bone. Interestingly, the changes in bone turnover induced by orchidectomy are paralleled by a concomitant increase in B lymphopoiesis in bone marrow of rats and mice. Although there is firm evidence that male bone metabolism can be influenced by androgens and estrogen, a variety of clinical and animal experimental data have strongly suggested that, under physiological circumstances, the maintenance of cancellous bone mass in males involves the skeletal action of estrogen derived from aromatization of androgens. Aged male rats appear to closely mimic the conditions induced by androgen withdrawal in adult humans, and this animal model may be used 1) to elucidate further the role of muscle as a mediator of the actions of androgens on bone, 2) to explore the regulatory functions of androgens and estrogens in the male skeleton and the immune system, and 3) to find new treatment strategies for the prevention and treatment of osteoporosis in men. PMID- 15758497 TI - Effect of circular motion exercise on bone modeling and bone mass in young rats: an animal model of isometric exercise. AB - The aims of the study are to develop a non-invasive animal model of circular motion exercise and to evaluate the effect of this type of exercise on bone turnover in young rats. The circular motion exercise simulates isometric exercise using an orbital shaker that oscillates at a frequency of 50 Hz and is capable of speeds from 0-400 rpm. A cage is fixed on top of the shaker and the animals are placed inside. When the shaker is turned on, the oscillatory movement should encourage the animals to hold on to the cage and use various muscle forces to stabilize themselves. Rats at 8 weeks of age were trained on the shaker for 6 weeks and static and dynamic histomorphometric analyses were performed for the proximal tibial metaphysis and the tibial shaft. The exercise resulted in no significant effect on animal body weight, gastrocnemius muscle weight and femoral weight. Although the bone formation rate of cancellous and cortical periosteum was increased by the exercise, trabecular bone volume was decreased. The exercise increased periosteal and marrow perimeters and the cross-sectional diameter of cortical bone from medial to lateral without a significant increase in the cortical bone area. These results suggest that circular motion exercise under force without movement or additional weight loading will cause bone-modeling drift with an increase in bone turnover to reconstruct bone shape in adaptation to the demand in strength. Since there is no additional weight loading during circular motion exercise, the net mass of bone is not increased. The bone mass lost in trabecular bone could possibly be due to a re-distribution of mineral to the cortical bone. PMID- 15758498 TI - A novel method to 'exercise' rats: making rats rise to erect bipedal stance for feeding - raised cage model. AB - We employed a novel method to exercise rats: making them rise to bipedal stance for feeding using raised cages. We studied its effects on the skeletons of 6 and 10-month-old intact or orchidectomized (ORX) rats. Body and hindlimb muscle weights, tibial BMC and periosteal cortical bone formation increased after housing in raised cages, but more so in 6-month-old animals than in 10-month-old ones. In 6-month-old orchidectomized rats, raised cages partially prevented ORX induced bone loss by stimulating periosteal cortical bone (TX) formation and decreased bone resorption next to marrow. In 10-month-old male orchidectomized rats, raised cages also decreased the endosteal and trabecular bone resorption, but not enough to prevent completely ORX-induced net bone losses. Because the osteogenic effects of raised cages alone were only partial, we also studied the interaction between raised cage and prostaglandin E(2) (PGE(2)) in 10-month-old retired female breeders. When treated with combined raised cage and PGE(2), both cortical (TX) and trabecular bone mass of the proximal tibial metaphysis and lumbar vertebral body increased over either raised cages or PGE(2) treatment alone, that was accompanied by dramatic increased bone formation at periosteal and endosteal surfaces. Thus making rats rise to erect bipedal stance for feeding helps to prevent bone loss after orchidectomy; it amplifies the anabolic effects of PGE(2), and it provides an inexpensive, non-invasive and reliable way to increase mechanical loading of certain bones of the rat skeleton. PMID- 15758499 TI - Skeletal loading in animals. AB - A number of in vivo skeletal loading models have been developed to test specific hypotheses addressing the key mechanical and biochemical signals involved in bone's adaptive response to loading. Exercise protocols, osteotomy procedures, loading of surgically implanted pins, and force application through the soft tissues are common approaches to alter the mechanical environment of a bone. Although each animal overload model has a number of assets and limitations, models employing extrinsic forces allow greater control of the mechanical environment. Sham controls, for both surgical intervention (when performed) and loading, are required to unequivocally demonstrate that responses to loading are mechanically adaptive. Collectively, extrinsic loading models have fostered a greater understanding of the mechanical signals important for stimulating bone cells, and highlighted the roles of key signaling molecules in the adaptive response. PMID- 15758500 TI - Analysis of biomechanical effects on bone and on the muscle-bone interactions in small animal models. AB - Animal models are suitable to study many aspects of bone structure and strength. This article reviews some general principles of current bone biomechanics and describes the scope of the available methodology for biomechanical studies of the musculoskeletal system employing those models. The analysis comprises bone and muscle "mass" indicators provided by standard densitometry (DEXA); bone 'mass', 'apparent density', geometry or architectural design and strength and muscle strength indicators that can be determined by peripheral quantitative computed tomography (pQCT), and bone material and structural (whole-bone) properties than can be directly assessed by destructive mechanical tests. Some novel interrelationships that can be investigated that way are discussed, namely, 1. the pathogenetic analysis of the effects on whole-bone strength, 2. the discrimination between mineralization and microstructural factors as determinants of changes in the bone material or structural properties, 3. the evaluation of the interaction of a treatment with the ability of bone 'mechanostat' to optimize the bone architectural design by 'distribution / mass' and 'distribution / quality' curves, and 4. the analysis of effects on the musclebone interactions for a differential diagnosis between 'physiological' or 'disuse' and 'true' osteopenias and osteoporoses. PMID- 15758501 TI - Transgenic and knock out mice in skeletal research. Towards a molecular understanding of the mammalian skeleton. AB - Our understanding of the biology of the skeleton, like that of virtually every other subject in biology, has been transformed by recent advances in human and mouse genetics. Among mammals, mice are the most promising animals for this experimental work. Because extensive genetic information exists, many mouse mutations are known, and cells from early mouse developmental stages are accessible, scientists have developed transgenic mice - mice in which a gene is introduced or ablated in the germ line. Thus far, we have analyzed more than 100 different transgenic and knock out models with various skeletal phenotypes, covering the major aspects of both skeletal development and skeletal maintenance. Based on these results we here present a first perspective on transgenic and gene knock out animals in skeletal research, including insights in signaling pathways controlling endochondral bone formation, in the regulation of osteoblast function, osteoclastic bone resorption and in bone tumorigenesis, as well as the central control of bone formation. The use of transgenic mice to dissect and analyze regulatory mechanisms in bone cell physiology and the pathogenesis of human bone diseases is an extremely powerful experimental tool. The data presented here demonstrate that the successful convergence of novel genetic approaches with the established and fundamental knowledge of bone biology has made a beginning. PMID- 15758502 TI - Anabolic agents and osteoporosis: quo vadis? AB - There are preclinical studies and limited clinical experiences with bone and muscle anabolic agents (e.g., parathyroid hormone (PTH), sodium fluoride (NaF), prostaglandins (PGs), growth hormones (GH), etc.) that show they have significant advantages over antiremodeling agents in patients with established osteoporosis. The strength of anabolic therapy is as follows: it rapidly reverses bone loss in laboratory animal models and humans, the quality of bone with some agents is believed to be normal, an increase in bone strength in animal models, and a reduction of spinal fracture rate with PTH. The weaknesses of this therapy are high cost, poor understanding of mechanism of action, parenteral mode of administration, rapid bone loss following termination of treatment, abnormal quality of bone, lack of tissue specificity, and undesirable side effects. Both animal and clinical studies have shown one can preserve the bone gain following termination of treatment with antiremodeling agents or exercise based on the lose, restore and maintain (LRM) concept. However, the more important efficacy issues which need to be addressed are tissue specificity and reduction of undesirable side effects. This report will address these issues with the suggestions that the potentiation of the mechanical loading osteogenic response by anabolic agents can overcome the disadvantages which accompany the use of anabolic agents. In addition, the possible role of nitric oxide (NO), an agent required for mechanical loading-induced bone formation, will be discussed. PMID- 15758503 TI - The Frozen Shoulder syndrome plus other evidence and the Utah Paradigm suggest the syndrome's pathogenesis and new targets for collagenous tissue research. AB - Sometimes naturally occurring disorders combined with other evidence can provide a "virtual laboratory" exercise that answers old questions and reveals new problems and questions that need appropriately "targeted" research. This article provides an example that involves the frozen shoulder syndrome, for which varied evidence and insights of the still-evolving Utah paradigm of skeletal physiology suggest a plausible pathogenesis. It involves organ- and tissue-level vital biomechanical features of collagenous tissues that need systematic study. The features include diametric modeling, irreversible creep, a creep compensation mechanism, the regional acceleratory phenomenon, and the signals and thresholds that can help to determine how and where those things act in anatomical space. Such features participate in many extraskeletal as well as skeletal disorders, so the frozen shoulder syndrome can provide an instructive model of them. As an aside, in over 120 consecutive cases managed by me since 1956 this syndrome resolved completely in six or less months without treatment. That shows this untreated syndrome is usually if not always self limited. While I respect the doubts of colleagues who do not agree yet, that experience shows the conclusion is correct. PMID- 15758504 TI - Parathyroid hormone and plasma calcium control: an editorial. AB - The purpose of this report is to examine the various processes by which parathyroid hormone might control the ionic calcium concentration of plasma and extracellular fluid, and to emphasize the need for study of the maintenance of plasma calcium in the absence of the parathyroid glands. The report discusses mechanisms to explain the control of extracellular calcium and proposes new approaches to the study of calcium homeostasis. PMID- 15758505 TI - Raloxifene: results from the MORE study. AB - Raloxifene is the first Selective Estrogen Receptor Modulator (SERM) approved for the prevention and treatment of osteoporosis in postmenopausal women. Acting as an estrogen agonist in the skeleton and on lipid metabolism, raloxifene maintains bone mineral density (BMD) and prevents new vertebral fractures while improving the lipid profile in postmenopausal women. In an osteoporosis prevention study, 601 women without osteoporosis, aged 45 to 60 years, were assigned to receive a placebo or raloxifene 30, 60, or 150 mg/day. All women received calcium (400 to 600 mg/day). Raloxifene 60 mg increased BMD by 2.4% at both the lumbar spine and hip compared with the placebo at 36 months. More importantly, however, raloxifene significantly reduced the risk of new vertebral fractures in Multiple Outcomes of Raloxifene Evaluation (MORE), a placebo-controlled, double-blind randomized trial of 7705 postmenopausal women with osteoporosis. The women, with a mean age of 66.5 years, and with hip or spine T-score <-2.5 and/or prevalent vertebral fractures, were assigned to receive either a placebo or 60 mg or 120 mg of raloxifene. All women were provided supplemental calcium (500 mg/day) and vitamin D (400 IU/day). After 36 months, raloxifene 60 mg/day and 120 mg/day, reduced the risk of new vertebral fractures by 55% (RR 0.45, 95% CI 0.3, 0.7; p<0.001), and 40% (RR 0.60, CI 0.4, 0.9) in women without prevalent baseline fractures, respectively; and by 31% (RR 0.7, 95% CI 0.6, 0.9; p<0.001), and 49% (RR 0.5, CI 0.4, 0.6) in women with prevalent baseline fractures compared with the placebo. There was no difference in the proportion of women reporting non-traumatic, non - spine fractures among women receiving raloxifene compared to the placebo-treated women. Compared with placebo, BMD increased after 36 months by 2.1 and 2.6% at the femoral neck and spine, respectively, in the 60mg raloxifene group, and by 2.4 and 2.7% at the femoral neck and spine, respectively, in the 120mg raloxifene group. By 40 months of follow-up, there was a higher rate of deep venous thrombosis (38 cases) and pulmonary embolus (17 cases) in the combined raloxifene groups than in the placebo group (5 and 3 cases,), with a relative risk of 3.1, (CI 1.5-6.2). By 40 months, 54 women had a confirmed diagnosis of breast cancer with a relative risk compared to placebo of 0.35, (CI, 0.21-0.58). Raloxifene therapy for 3 years maintains BMD in healthy postmenopausal women and significantly reduces the risk of new vertebral fractures by about half in postmenopausal women with osteoporosis. Raloxifene also reduces the risk of breast cancer by 65% in postmenopausal women with osteoporosis thus providing a new choice for addressing postmenopausal health concerns. PMID- 15758506 TI - The osteocyte as a wiring transmission system. AB - The mechanism of transduction of mechanical strains into biological signals remains one of the more baffling problems of skeletal homeostasis. The updated literature ascribes to osteocytes the function of sensing the strains induced into the bone matrix by mechanical stresses. Whether the osteocytes perform such function by themselves or they are helped by other cells is also unknown. Indeed TEM investigations carried out in our laboratory pointed out the existence of a functional syncytium among all the cells of the osteogenic lineage (COL; stromal cells, osteoblasts or bone lining cells, osteocytes). On the basis of this finding, we suggested that COL may reciprocally modulate their function not only by volume transmission (paracrine and autocrine stimulation) but also by wiring transmission, namely in a neuronal like manner. Thanks to their location, osteocytes should theoretically be the first cells of COL functional syncytium to sense mechanical strains, whereas stromal cells should be the first to be activated by hormonal molecules diffusing across the endothelial lining. Since PTH and Estrogen receptors have also been localized on osteocytes, and considering that such hormones have been suggested to modulate the sensitivity to strain of the bone mechanosensor, we suggested that the osteocyte syncytium may constitute the microscopic bone structure that sense both mechanical strain and biochemical factors and, at any moment, after having combined the two types of stimuli, issues the appropriate signals to the other bone cells by volume and/or wiring-transmission. Stromal cells, on the other hand, besides transmitting signals from vascular endothelium to bone cells, may control the differentiation and then direct the course of the osteoblasts around the vascular framework. PMID- 15758507 TI - Some vegetables (commonly consumed by humans) efficiently modulate bone metabolism. AB - We have hypothesized that some vegetables which are part of the regular human diet may contain modulators of bone metabolism. To mimic a typical Western diet with large proportions of refined components, rats were pair-fed a semi-purified diet to which, in the treated animals, the dried material under investigation was added. Effects are expressed as % of untreated control. Bone parameters in rats were assessed in the proximal tibia by pQCT. Bone resorption (BR) was assessed by the urinary excretion of [3H]-tetracycline from prelabeled rats. Daily administration of 1 g of onion during 4 weeks increased total bone mineral content by 17.4% (p<0.05), trabecular bone mineral density by 13.6% (p<0.05). One g of onion/day administered to male rats blunted BR by 23-/+5% (p<0.05). Daily administration of onion to ovariectomized rats inhibited BR in a dose-dependent manner. At the highest dose (1.5 g of onion) BR was inhibited by 26-/+4% (p<0.01) as compared to 24-/+3% (p<0.001) for estradiol (27microg/kg/day). An additional 13 vegetables displayed significant effects on BR at the dose of 1g/day. Interestingly, 1g/day of soy did not inhibit BR in this model. Also, skimmed milk, meat and egg (all 1 g/day) were ineffective. Thus, common vegetables consumed by humans potently modulate bone metabolism in the rat. This opens the possibility to develop the basis for a low-cost, safe and effective nutritional approach to osteoporosis. PMID- 15758508 TI - The mechanism of bone resorption by cyclosporin: involvement of the NO-cGMP pathway. AB - Treatment with cyclosporin A (CsA) following solid organ transplantations such as heart or liver generally results in bone loss. However, in vitro studies show that CsA inhibits bone resorption. Our previous in vivo animal studies demonstrated that the effects of nitric oxide (NO) on bone are biphasic; at high doses, NO increases bone resorption. In this study, we have examined in an in vitro setting to determine whether the bone loss caused by CsA administration is dependent on the NO-cyclic guanosine monophosphate (cGMP) pathway. Freshly isolated osteoclast-rich neonatal rat long bone marrow cells were added to 100 microM thick dentin sections that had been seeded with neonatal-rat calvarial osteoblasts. These co-cultures were maintained for 48 hrs in a basal medium with CsA (1, 5, and 10 microg/ml), both alone and with either L-Arginine (NO substrate; 10-3M), L-NAME (NO synthase enzyme inhibitor; 10-4M), or the combination of the two. The cultures were then fixed in cold 95% ethanol and stained with tartrate resistant acid phosphatase (TRAP) to identify osteoclasts and sites of osteoclastic resorption. Preparations were analyzed using an automated histomorphometry software package. Scanning electron microscopy affirmed that the areas identified by light microscopy as resorption sites contained osteoclastic lacunae. CsA inhibited bone resorption dose-dependently. CsA at 10 microg/ml produced a 90% inhibition of bone resorption (control = 5.5 /+2.0%; CsA = 0.64 -/+ 0.09=). L-Arginine reversed this inhibition by 90% (Arg + CsA = 4.23 -/+ 1.57%; CsA = 0.64 -/+ 0.09%). The application of NOS inhibitor L NAME inhibited bone resorption by 87% (Arg + CsA + L-NAME = 0.55 -/+ 0.14%; Arg + CsA = 4.23 -/+ 1.5%). We conclude that NO-cGMP pathway is involved in the CsA induced bone loss. PMID- 15758509 TI - Effects of racetrack exercise on third metacarpal and carpal bone of New Zealand thoroughbred horses. AB - The response of equine bone to training has not been quantified in racetrack trained horses, only in treadmill exercised horses. Seven two-year-old thoroughbred fillies were trained on sand and grass at a racetrack, in a typical New Zealand flatrace training regime. The horses were exercised 6 days per week for up to 13 weeks. During the day the horses were confined in 4 x 4m sand yards, and were stalled at night. Another 7 fillies of the same age were allowed free exercise in grass yards. The bones of the animals were available after the 13 week experimental period, and were examined using a Siemens Somatom AR CT scanner. To quantify the response of epiphyseal bone, 3mm thick sagittal plane images of the carpus (through the middle of the medial condyle of distal radius) and the distal third metacarpal bone (Mc3) (immediately lateral and medial to the junction of the condyle and the median sagittal ridge) were studied. Appropriate areas of interest were chosen, and the mean tissue density equivalent (Houndsfield Units) was determined. In the carpus, there was a significant effect of exercise in the dorso-distal aspect of the radius (p<0.01), dorsal aspect of radial and third carpal bones (p<0.01 and p<0.001 respectively). In palmaro distal subchondral bone of Mc3, there was a significant effect on the medial/lateral site (p<0.01), which differed between right and left legs, probably due to the effect of the horses having been trained in one direction around the training track. The mean tissue density of the Mc3 epiphysis of the exercised group was 36.8% greater than that of the non-exercised group (p<0.001). The study demonstrates that bone response is both rapid and substantial, which should prompt the use of non-invasive diagnostic aids to determine the stage of training in which tissue density changes occur. PMID- 15758510 TI - Long-lasting effect of pamidronate on bone metabolism in osteoporosis after stopping therapy. AB - AIM OF THE STUDY: To determine whether and when, like after estrogen withdrawal, bone loss resumes once disodium pamidronate is discontinued, even after long duration therapy. MATERIALS AND METHODS: 19 patients with osteoporosis, previously treated for 4.3 -/+ 0.5 years (SEM) with oral cyclical intermittent pamidronate, were followed-up for 30.3 -/+ 2.2 months after withdrawal from therapy. Lumbar spine and proximal femur BMD was measured by DXA (QDR-1000, Hologic Inc., Waltham, MA). RESULTS: Lumbar spine BMD did not change significantly for the first 2 years, but decreased 1.8 % in the 3rd year (p<0.001). The proximal femur BMD did not change significantly for 3 years. The biological parameters of bone remodelling increased progressively with elapsing time, due to the underlying loss of pamidronate protection. CONCLUSIONS: after withdrawal of pamidronate therapy, a residual protecting effect was observed at the proximal femur and at the lumbar spine for 2 to 3 years. The time-interval resurgence of bone remodelling and the bone loss observed at the spine in the 3rd year suggest that there is no risk of freezing bone with pamidronate therapy, even at high doses. A long-lasting protective effect on bone mass can be expected. Periods of therapy with an active drug, interrupted by long resting periods, may produce the same protective effect as continuous therapy, owing to the levelling-off effect on bone mass observed after the first two years while on therapy. This would lead to lower expenses for a course of therapy. PMID- 15758511 TI - Three-dimensional type I collagen co-culture systems for the study of cell-cell interactions and treatment response in bone metastases. AB - The available monolayer culture systems for the study of bone metastases constitute a suboptimal simulation of the in vivo pathophysiology of bone metastases, and therefore, do not provide sufficient information to assess the morphologic evidence of bone reaction to cancer cells, the nature of cell specific mediators of osteolysis and osteoplasia and the response to treatment. Therefore, we have developed a three-dimensional (3-D) type I collagen gel system that allows co-culture of human osteoblasts (MG-63) with cancer cells, such as MCF-7, MDA-MB-231 or ZR-75 breast cancer cells, PC-3 prostate cancer, KLE endometrial cancer cells and Calu-1 lung cancer cells. We used type I collagen purified from rat tail tendons and the 3-D system was prepared by mixing MG-63 cells with type I collagen in 24-well plates. The 3-D system was inoculated with cancer cells and processed with standard cell culture procedures. After 1 week of culture, the matrix gel was fixed with formalin and embedded in paraffin. Serial sections were stained with trichrome Masson stain and modified Masson-Goldner stain, as well as analyzed by in situ hybridization, immunohistochemistry and the TUNEL technique for semi-quantitative detection of apoptotic cell death, assessing the response to adriamycin therapy. The inoculation of PC-3 cells in this collagen matrix produced a blastic reaction, documented by an increased number of MG-63 cells and increased density of type I collagen. The human KLE cells and inoculation of cell-free media produced no reaction, while ZR-75, MCF-7 and Calu-1 cells produced local degradation of the collagen matrix. In situ hybridization revealed the expression of Insulin-like growth factor 1 (IGF-1) and urokinase-type plasminogen activator (uPA) mRNA, while immunohistochemistry detected differential expression of uPA and cathepsin D. Adriamycin induced apoptotic cell death in prostate cancer cells and estrogen receptor negative (ER ) MDA-MB-231 breast cancer cells, while adriamycin did not induce apoptosis but cytostasis in ER+ MCF-7 cells. The adriamycin-induced apoptosis was inhibited by co-culture with osteoblast-like cells (MG-63). We conclude that this 3-D culture system is a useful in vitro model allowing the analysis of local mediators of osteolytic and osteoblastic reactions to bone metastases and treatment response. PMID- 15758512 TI - Bone mineral and lean tissue loss after long duration space flight. AB - The loss of bone and muscle is a major concern for long duration space flight. In December of 1989, we established a collaboration with Russian colleagues to determine the bone and lean tissue changes in cosmonauts before and after flights on the Mir space station lasting 4-14.4 months. Eighteen crew members received a lumbar spine and hip DEXA scan (Hologic 1000W) before and after flight; 17 crew members received an additional whole body scan. All results were expressed as percent change from baseline per month of flight in order to account for the different flight times. The pre-and post-flight data were analyzed using Hotelling's T(2) for 3 groups of variables: spine, neck of femur, trochanter; whole body BMD and subregions; lean (total, legs, arms) and fat (total only). A paired t-test was used as a follow-up to the Hotelling's T(2) to identify the individual measurements that were significantly different. These data define the rate and extent of bone and lean tissue loss during long duration space flight and indicate that the current in-flight exercise program is not sufficient to completely ameliorate bone and muscle loss during weightlessness. PMID- 15758513 TI - In vitro effects of dynamic strain on the proliferative and metabolic activity of human osteoblasts. AB - AIM OF THE STUDY: It has been well shown by human and animal studies that mechanical load is an important regulator of skeletal mass and architecture. However, cellular reactions which adapt bone tissue to the mechanical environment are not definitively determined. For this purpose we studied the cell activity of human bone derived cell cultures after mechanical stimulation by cyclic, uniaxial strain at a magnitude occurring in normal loaded bone tissue. MATERIALS AND METHODS: Human osteoblasts were isolated from cancellous bone biopsies of 5 different donors. Cell seeding was made in DMEM in a density of 10.000 cells/cm(2) on deformable culture dishes for three days prior to initiating cell stretching at 1000 microstrain, 1Hz for 1800 cycles for two subsequent days with an especially developed cell stretching device. 48h after the second stimulation cells were harvested and cell number was determined with a Coulter Counter. Cell bound alkaline phosphatase activity was analyzed in cell lysates by a colorimetric assay, osteocalcin and CICP (procollagen I propeptide) production were analyzed in cell supernatants with ELISAs. Three parallel cultures were tested. STATISTICS: Wilcoxon. RESULTS: In all experiments mechanical stimulation resulted in a significant increase in cell number (10-48%) and CICP release (7 49%). Simultaneously a significant decrease in alkaline phosphatase activity (9 25%) and osteocalcin release (5-32%) could be observed. CONCLUSIONS: The results demonstrate that cyclic strain at physiologic magnitude leads to an increase of early osteoblast activities related to matrix production while those activities which are characteristic for the differentiated osteoblast and relevant for matrix mineralization are decreased. These new findings confirm in vivo observations about the importance of dynamic strain for bone formation during fracture healing and bone remodeling and could contribute to the optimization of fracture healing. PMID- 15758514 TI - Evaluation of threshold stress for bone resorption around screws based on in vivo strain measurement of miniplate. AB - The purpose of this study is to investigate the critical threshold stress causing bone resorption evaluated from strain measurement in vivo, comparing the various finite element models. In this study strains of miniplates used for mandibular fractures were measured once a week until the strains reduced. The maximum bite force for each patient was applied in the incisal, right molar and left molar region. The strains increased and reached a peak level at 2-4 weeks, whereas the bite forces increased during the period of measurements. A 3-D osteosynthesis model using finite element method showed that the compressive stresses of the bone surrounding screws ranged within approximately -40 MPa under the condition generating the same amounts of strains measured in the miniplates. Furthermore, various finite element models simulating mandibular reconstruction using the fibular graft were constructed. The models for reconstruction using single strut fibula showed distinct stress concentration in the cortical bone surrounding screws, and the peak stress levels were 2 to 3 times as strong as that of the fracture model. We conclude that critical threshold for bone resorption should be approximately -50 MPa (3600 micro strain). PMID- 15758515 TI - What is new in musculo-skeletal interactions. AB - In this new column of the JMNI, we will give a six-monthly overview about new publications which, to our eyes, are interesting and relevant. Keeping in line with the 'spirit' of ISMNI, we take the freedom to discuss with a more holistic approach and most importantly, we encourage feed-back from the readers. PMID- 15758516 TI - The history of the walls of the Acropolis of Athens and the natural history of secondary fracture healing process. AB - During its long and adventurous history, the Acropolis of Athens has been a site of many dramatic events. It suffered its most disastrous destruction during the Persian wars. Under the command of King Xerxes, the Persians invaded Athens and ruined the Temple of the Parthenon and the walls of the Acropolis. After their victorious sea battle at Salamis, the Athenians, led by Themistocles, returned home and tried to repair the damage. Their priority still was to defend their city by restoring the walls of the Acropolis. Materials of all kinds were salvaged from the ruins of the Acropolis and used for an immediate reconstruction of the walls. Later, when the Athenians became the leaders of the Greek world, it was decided that the walls should be rebuilt in a proper artistic way. Themistocles suggested that a small section of the walls, which had formerly been a part of the urgent restoration, should remain in place so as to remind the citizens of this historical event. This is a characteristic example of the biological and mechanical adaptation of fracture callus to musculoskeletal function. After a period of urgency with the fixation of a fracture by means of a primitive secondary callus formation, the broken limb gradually returns to its usual function. Increased mechanical loading enhances the remodelling of the callus and the replacement of woven bone with lamellar bone. PMID- 15758517 TI - Why the ISMNI and the Utah paradigm? Their role in skeletal and extraskeletal disorders. AB - Besides bringing problems, aging can let the mind's eye see more clearly than before, and it can let us express ourselves better. As age, experience and common sense examine today's skeletal medicine and surgery two questions keep popping up: A) How did we fail?; B) How to make it better? The Utah paradigm of skeletal physiology and the seminal ISMNI offer some answers, but exploiting them faces problems. Problem #1: By 1960 all clinicians and physiologists 'knew' (as the ancients 'knew' this world is flat) that effector cells controlled solely by nonmechanical agents explain all skeletal physiology and disorders ('effector cells' include osteoblasts, osteoclasts, chondroblasts and fibro-blasts). Or, nonmechanical agents -->cell level -->organ and intact subject. Adding later discovered information to that 1960 view led to the Utah paradigm, which reveals the formerly hidden tissue-level 'dimension' of skeletal physiology. It builds on this idea: (mechanical + nonmechanical agents) -->(tissue level + cell level) --> organ and intact subject. The paradigm assigns great influence of neuromuscular physiology and physical activities on skeletal architecture, strength and mechanical competence. It also exposes flaws in many older views so controversies arise. Problem #2: The Utah paradigm and Wegner's concept of plate tectonics in geology seem alike in that each is valid but came before its time, so others fought it. They differ in this: The fight about Wegner's idea is over, but for the Utah paradigm and the ISMNI it just began. Hence more controversies. Nevertheless: A growing minority realizes that paradigm provides a far better base to build on than its antecedents, and since it keeps evolving as more evidence comes in it could endure for some decades. Yet very few realize this: It and the ISMNI have important implications for fields besides biomechanics and orthopaedics. Examples include anatomy, cardiovascular disease, dentistry, endocrinology, family medicine, gastroenterology, general surgery, genetics, gerontology, gynecology, maxillofacial surgery, neurology, neurosurgery, nutrition, ophthalmology, pathology, pediatrics, physical medicine and rehabilitation, plastic surgery, radiology, rheumatology, space and sports medicine, and urology. Quite a list! For the italicized questions above this article offers answers, of which its conclusion distills an essence. PMID- 15758518 TI - Principles in bone physiology. AB - The view that nonmechanical agents dominate control of osteoblasts and osteoclasts and thus postnatal changes in bone strength and mass (agent --> effector cells --> disease) is obsolete. Nonmechanical agents include hormones, calcium, vitamin D, cytokines, gender, genetics, etc. This paradigm overlooks all tissue level features, biomechanics and relationships found after 1960. This more recent information led to the Utah paradigm of skeletal physiology, proposed by Harold Frost in 1995. The Utah paradigm's view is that mechanical factors dominate control of the biologic mechanisms that control changes in postnatal bone and mass. Nonmechanical agents could help or hinder the influence of the mechanical factors but could not replace them. The simplified scheme is as follows: [reaction: see text] New evidence supports the Utah paradigm which we view as a supplement to many former views, not as a negation of them. PMID- 15758519 TI - Cancer and bone repair mechanism: clinical applications for hormone refractory prostate cancer. AB - It is a long-standing clinical observation that the bone corresponds to the prevalent site for metastatic growth of prostate cancer. In addition, bone metastases of this malignancy produce a potent blastic reaction, in contrast to the overwhelming majority of other osteotropic neoplasms, whose metastases are generally associated with an osteolytic reaction. Osteoblastic metastases represent almost always the first and, frequently, the exclusive site of disease progression to hormone refractory stage, stage D3. Moreover, the number of skeletal metastatic foci is the most powerful independent prognostic factor associated with a limited response to hormone ablation therapy and poor survival of advanced prostate cancer. It is noteworthy that disease progression to hormone refractory stage occurs almost always in osteoblastic metastases. These clinical observations suggested that the osteoblastic reaction is possibly not an innocent bystander of the metastatic prostate tumour growth, simply suffering its consequences, but it may in fact facilitate the efforts of metastatic cells to expand their population. An extensive line of research in the pathophysiology of osteoblastic metastases has established that the local blastic reaction involves the uPA/plasmin/IGF/IGFBP-3/TGFbs bioregulation system which can stimulate both the growth of osteoblasts and prostate cancer cells. Furthermore, we were the first to characterize osteoblast-derived 'survival factors' able to rescue metastatic prostate cancer cells from chemotherapy-induced apoptosis. These data resulted in the development of a novel concept of an anti-survival factor therapy, namely an anti-IGF-1 therapy, which has provided encouraging preliminary data in a phase II clinical trial with terminally-ill hormone/chemotherapy resistant prostate cancer patients. PMID- 15758520 TI - How cancellous and cortical bones adapt to loading and growth hormone. AB - There is great interest in the relationships between growth hormone (GH), muscle loading and bone, in part, because GH increases muscle mass which provides the largest signals that control bone modeling and remodeling. This study was designed to examine the effects of GH and muscle loading by exercise (EX) independently and in combination on bone and skeletal muscle. Thirteen-month-old female F344 rats were divided into 6 groups: Group 1, baseline controls (B); Group 2, agematched controls (C); Group 3, GH treated (2.5 mg rhGH/kg b. wt/day, 5 days per week); Group 4, voluntary wheel running exercise (EX); Group 5, GH+EX, and rats in Group 6 were food restricted (FR) to lower their body weight and examine the effects of decreased muscle load on bone. All animals, except the baseline controls, were sacrificed after 4.5 months. Growth hormone increased the body weight and tibial muscle mass of the rats markedly, while EX caused a slight decrease in body weight and partially inhibited the increase caused by GH in the GH+EX group. Food restriction greatly decreased body weight below that of age matched controls but neither FR nor EX had a significant effect on the mass of the muscles around the tibia. Growth hormone and EX independently increased tibial diaphyseal cortical bone area (p<0.0001), cortical thickness (p<0.0001), cortical bone mineral content (p<0.0001), periosteal perimeter (p<0.0001) and bone strength-strain index (SSI) (p<0.0001). The effects of GH were more marked, and the combination of GH and EX produced additive effects on many of the tibial diaphyseal parameters including bone SSI. GH+EX, but not GH or EX alone caused a significant increase in endocortical perimeter (p<0.0001). In the FR rats, cortical bone area and cortical mineral content increased above the baseline level (p<0.0001) but were below the levels for age-matched controls (p<0.0001). In addition, marrow area, endocortical perimeter and endocortical bone formation rate increased significantly in the FR rats (p<0.01, p<0.0001, p<0.0001). Three point bending test of right tibial diaphysis resulted in maximum force (Fmax) values that reflected the group differences in indices of tibial diaphyseal bone mass except that GH+EX did not produce additive effect on Fmax. The latter showed good correlation with left tibial diaphyseal SSI (r=0.857, p<0.0001) and both indices of bone strength correlated well with tibial muscle mass (r=0.771, Fmax; r=0.700, SSI; p<0.0001). We conclude that the bone anabolic effects of GH with or without EX may relate, in part, to increased load on bone from tibial muscles and body weight, which were increased by the hormone. The osteogenic effects of EX with or without GH may relate, in part, to increased frequency of muscle load on bone as EX decreased body weight (p<0.05) but had no significant effect on tibial muscle mass. The enhanced loss of endocortical bone by FR may relate, in part, to decreased load on bone due to low body weight (p<0.0001) as FR did not cause a significant decrease in tibial muscle mass (p=0.357). The roles of humoral and local factors in the bone changes observed remain to be established. PMID- 15758521 TI - Trabecular bone structure and strength - remodelling and repair. AB - The strength of the spinal trabecular bone declines by a factor of 4-5 from the age of 20 to 80 years. At the same time, the volumetric (apparent) density declines by a factor of only 2. This discrepancy can be explained by the known power relationship between density and strength; this power relationship is based on the fact that trabecular bone is a porous material. To date, it has not been possible to determine or quantify the influence other factors may have in determining the strength of a loadbearing trabecular network. However, it is known that with age: 1) There is a loss of connectivity through osteoclastic perforations of horizontal struts. 2) There is an increase in anisotropy - again due to loss of horizontal struts, and perhaps also due to micro-modelling drift or to thickening of some vertical trabeculae. 3) The changes in the network can lead to the slenderness ratio between vertical and horizontal struts reaching a certain magnitude and thereby inducing buckling under compression. 4) Microdamage and microfractures will occur - mainly in these very loaded vertical struts. The microfractures will be repaired by microcallus formation, and these calluses will later be removed by the remodelling process. 5) Bone material quality will slightly change, leading to a decrease in collagen content and a relative increase in the degree of mineralisation. But, it is not known how these factors will influence the power relationship between density and strength. Nor is it known how different treatment regimens will affect the 'natural' power relationship: will the same curve be followed, but in the opposite direction? Or will the curve be less or more steep? Will the gain in bone strength be larger if treatment is started early - on the steep part of the curve? Furthermore, as trabecular bone can never be isolated in vivo, other factors need to be investigated: The interplay between the cortical shell and the trabecular network; transmission of load; the interplay between soft tissues (cartilage, connective tissue, muscle) and bone; the shock absorbing capacity of the discs; and the hydraulic effect of the bone marrow. In order to answer these questions, more in vitro and in vivo studies on human bone in relation to aging, to immobilisation, to exercise and in relation to different treatment regimens are needed. PMID- 15758522 TI - Densitometric and tomographic analyses of musculoskeletal interactions in humans. AB - Previous studies with standard densitometry (DXA) have suggested that the bone mass is strongly dependent on the muscle mass in the species, following a similar relationship at any age and sex hormones or related factors potentiate that relationship. Studies with pQCT indicated that the surplus bone mass per unit of muscle mass previously observed in premenopausal women would be stored in skeletal regions with relatively little mechanical relevance, thus avoiding remotion through mechanically oriented remodelling by the bone mechanostat. Scanning the distal radius with pQCT has also showed a highly significant, linear relationship between SSI of the distal radius and the dynamometric maximal bending moment of the forearm in normal men and women. In order to investigate similar relationships in regions that are inaccessible to pQCT, we used spinal radiographs and axial QCT. This study affords additional evidence to the previous references concerning the direct, significant impact of the regional muscle strength on the determination of the tomographic indicators of bone mechanical quality and their indirect repercussion of the skeletal condition (curvature of the spine). PMID- 15758523 TI - A novel pharmacological approach of musculoskeletal losses associated with simulated microgravity. AB - Exposure to microgravity (weightlessness) is known to cause rapid bone and muscle losses. We have used the hind limb-suspended (HLS) rat model to simulate microgravity-induced musculoskeletal losses in order to assess resulting hormonal changes and to develop a novel pharmacological countermeasure. Previously, we demonstrated significant decreases in circulatory hormonal levels [serum thyroxin, 1,25(OH)2 vitamin D (p<0.05), and serum testosterone (p<0.001)] in HLS rats. Both thyroxin and 1,25(OH)2 vitamin D levels returned to normal soon after removal from HLS, while testosterone levels matched normal levels only after a further 3-4 weeks. However, even by day 42, bone mineral density (BMD) remained significantly lower, although serum hormones were back to normal. Because serum testosterone levels become undetectable in HLS rats, we hypothesized that the replacement of testosterone during HLS could prevent musculoskeletal losses. Based on these data, an intervention study was carried out to assess the efficacy of testosterone and synthetic anabolic steroid, nandrolone decanoate (ND), in prevention of weightlessness-induced musculoskeletal losses. HLS rats (control) had a significant reduction of muscle volume (42.9 -/+ 3.0, versus 56 -/+ 1.8 in ground control rats; p<0.01). Both testosterone and ND treatments prevented this muscle loss (51.5 -/+ 2 cm(3) and 51.6 -/+ 1.2, respectively; a 63% improvement, p<0.05). Similarly, BMD of the placebo-treated HLS rats was significantly lower than that of ground control rats (0.416 -/+ 0.011 versus 0.354 -/+0.014, p<0.05), and testosterone and ND prevented this bone loss (0.404 -/+ 0.013 versus. 0.409 /+ 0.011, respectively). These data suggest that both testosterone and ND therapy can minimize the musculoskeletal losses associated with exposure to simulated weightlessness. Experiments using the combination of bisphosphonate and testosterone demonstrated complete protection of both muscle and bone in these HLS rats. Therefore, considering that: 1) testosterone is anabolic to osteoblasts and muscle cells and also decreases the rate of bone turnover, 2) serum testosterone levels are markedly suppressed in simulated weightlessness, and 3) testosterone replacement therapy prevented musculoskeletal losses in HLS rats, we propose that the musculoskeletal losses observed in this animal model (i.e., simulated microgravity) are related to their testosterone deficiency. Since serum sex hormones levels are markedly reduced in this model of simulated microgravity, androgen replacement with a bisphosphonate seems to be a rational counter. PMID- 15758524 TI - Essential amino acid supplements increase muscle weight, bone mass and bone strength in adult osteoporotic rats. AB - Protein undernutrition is known to play an important role in the pathogenesis of osteoporotic fracture in elderly. The mechanisms underlying the bone loss in protein undernutrition appeared to be related to an uncoupling between increased bone resorption and bone formation. This was associated with decreased plasma IGF I levels, with anoestrus and decreased muscle mass. Reversibility of protein undernutrition-induced bone loss was investigated in ovariectomized adult rats, which were fed isocaloric 2.5 % casein diet (OVX2.5) for 16 weeks. Then, the animals were given a supplement of essential amino-acids in similar proportion to that of casein at doses of 2.5% (EAA2.5) or 5% (EAA5) of total food intake for an additional 16 weeks. Essential amino acid supplements increased bone mineral mass and strength in ovariectomized protein-deprived rats. EAA supplements were associated with stimulated bone formation and reduced bone resorption, with increment of plasma IGF-I and of limb muscle mass weight. These results suggest that nutritional intervention with essential amino acid supplements can increase bone mineral mass, bone strength and muscle mass in osteoporotic rats possibly by correcting IGFI status. PMID- 15758525 TI - Micro CT and Micro MR imaging of 3D architecture of animal skeleton. AB - Quantitative assessment of three-dimensional (3D) trabecular structural characteristics may improve our ability to understand the pathophysiology of osteoporosis, to test the efficacy of pharmaceutical intervention, and to estimate bone biomechanical properties. Considerable progress has been made in advanced imaging techniques for noninvasive and/or nondestructive assessment of 3D trabecular structure and connectivity. Micro computed tomography (microCT) has been used to measure 3D trabecular bone structure in rats, both in vivo and in vitro. It can directly quantify 3D trabecular bone structure such as trabecular volume, trabecular thickness, number, separation, structure model index, degree of anisotropy, and connectivity, in a model-independent manner. We have used microCT to study ovariectomy (OVX) induced osteopenia in rats and its treatment with agents such as estrogen, and sodium fluoride. We have demonstrated that 3D microCT can quantify mouse trabecular and cortical bone structure with an isotropic resolution of 9 microm(3). It is also useful for studying osteoporosis in mice and in phenotypes of transgenic mice or gene knockout mice. MicroCT can be used to quantify osteogenesis in mouse Ilizarov leg lengthening procedures, to quantify osteoconduction in a rat cranial defect model, and to quantify cortical bone porosity. Recently, microCT using high intensity and tight collimation synchrotron radiation to achieve spatial resolution of 1-2 microm has provided the capability to assess additional features such as resorption cavities. Unlike microCT, micro magnetic resonance imaging (IMRI) is nonionizing. Recently, the ability of microMRI to assess osteoporosis in animal models has been explored. Using a small, high-efficiency coil in a high-field imager, microMRI can give resolutions sufficient to discriminate individual trabeculae. We have shown that, with appropriate settings, it is possible to image trabecular bone in rats in vivo and in vitro. In our study of OVX rats, analysis of microMR images can demonstrate differences in rat trabecular bone that are not detected by DXA measurements. In a rabbit OA model, with the OA induced by meniscectomy or anterior cruciate ligament transection, MRI shows decreased cartilage thickness, subchondral osteosclerosis and osteophytes, while radiographs can only show subchondral osteosclerosis and osteophytes could not be found. Advanced imaging methods are able to measure 3D trabecular structure and connectivity in arbitrary orientations in a highly automated, objective, non-user-specific manner, allowing greater numbers of samples for unbiased comparisons between controls and the disordered or treated. They can be utilized on a large sample leading to fewer sampling errors. They are non-destructive allowing multiple tests such as biomechanical testing and chemical analysis on the same sample; and non-invasive permitting longitudinal studies and reducing the number of animals needed. PMID- 15758526 TI - PTH and interactions with bisphosphonates. AB - We report that a therapeutic dose of the antiresorptive bisphosphonate alendronate administered to skeletally mature rats for the duration of 16 weeks significantly blunted the anabolic response to a high dose SDZ PTS 893 in the tibia and femur but not in lumbar vertebra. Effects were seen at the level of bone mass (DEXA, pQCT) as well as in biomechanical tests. In one arm of this study, rats were switched to vehicle injections after 8 weeks on alendronate for another 8 weeks before being challenged with the anabolic stimulus (washout). This recovery period was insufficient for full recovery and the response to SDZ PTS 893 was still greatly reduced after this procedure. Serial pQCT-measurements suggest that part of the interaction happened during the first two weeks of PTH treatment when bone-lining cells are activated by the anabolic drug. In addition bisphosphonate pretreated rats failed to catch up with the vehicle control at all time points suggesting a second level of drug interaction. The failure of the 'washout' period to restore the normal response to PTH is suggestive of a physico chemical interaction on the level of the matrix embedded bisphosphonate with the overlaying bone lining cells, rather than of direct effects of the drug on osteoblasts or their precursor cells. Overall the data raises the possibility, that bisphosphonate treated patients respond to PTH and SDZ PTS 893 with a delay which could affect the shorter bone mass measurements carried out at 6 months to 1 year. Additionally, bisphosphonate pre-treated rats did not develop the full anabolic response over time. Clinical investigators studying anabolic drugs such as PTH should be aware of potential long-term interactions of bisphosphonates when assessing the outcome of their experiments. However, the beneficial effect of bisphosphonates like alendronate on PTH-induced bone remodeling, as well as its potent action in the protection of bone loss after cessation of anabolic therapy might outweigh the worries about a small delay in the bone response to parathyroid hormone. PMID- 15758527 TI - Periprosthetic bone densitometry of the hip: influence of prosthetic design and hydroxyapatite coating on regional bone remodelling. AB - OBJECTIVE: To determine the influence of femoral hip prosthesis design and composition on regional periprosthetic bone remodelling as a function of time in asymptomatic patients. MATERIALS AND METHODS: Bone mineral density (BMD) changes surrounding two differently designed porous-coated femoral prostheses (S-ROM, n=111; and Multilocki, n=62) were determined serially over a 24 month interval. There was a subset of patients with Multilock implants that were coated with a 50 micron film of hydroxy-apatite (n=23) over the porous surface. Seven Gruen zones defined the periprosthetic bone regions (LUNAR ORTHO software). Inclusion criteria were primary implants, Harris hip scores >or=95 and no radiographic evidence of loosening. RESULTS: For the S-ROM femoral component, at 6 months all zones showed a significant decrease in BMD relative to the immediate postoperative baseline measurements; mean loss varied from 4.3% to 17.4%. At 24 months mineral change varied from -10.8% to +1% for Gruen zones 1 to 6. Gruen zone 7, the calcar, differed in that it was the site of greatest mineral loss, attaining a mean of 17% at 24 months, and it was also significantly greater than the losses registered in the other zones at that time point. For the Multilock, maximal mineral losses were registered at zones 1 and 7. There were significant differences in mineral losses between the S-ROM and Multilock implants in zones 1, 2 and 3 at 24 months; S-ROM losses of 7.3%, 10.8% and 3.5% respectively, compared to Multilock losses of 23.8%, 15.9% and 6.9% respectively. Comparison of hydroxyapatite coated and uncoated Multilock implants demonstrated significantly less loss in the coated implants in zones 1 (greater trochanter) and 7 at 24 months. Coated losses were -16.9% and 11.3% for zones 1 and 7, respectively, whereas uncoated losses were 23.8% and 20.5% respectively. PMID- 15758528 TI - Balance training and exercise in geriatric patients. AB - Objective measures of gait and balance which meet the criteria of reliability and validity are required as a basis for exercise regimens. We established reference values of clinically relevant locomotor and balance performances for geriatric patients. We are using these data for evaluating the effects of different therapeutic approaches to locomotor and balance disorders. Reference values for chair rising. We administered a battery of five tests concerning neuromuscular function, locomotion and balance to a sample of 212 participants without apparent locomotor deficits (139 women, 73 men, mean age 70,5 years, SD 6,78 , median 70 years, range 60 to 90 years, recruited by public announcements). The test battery comprised the 'chair rising test' for measuring lower extremity neuromuscular function (five repetitions of rising from a chair as quickly as possible with arms crossed over the chest). The test has been proven reliable, valid, sensible and predictive for falls and future locomotor status and ADL-status. Chair rising [sec/5x], Range: 5.4-19.4, Mean: 9.1 (women:9.2, men:9.0), SD: 1.97, Median: 8.9. Training of balance and muscle power with Galileo 2000 - preliminary results. Galileo is a device for whole body vibration/oscillatory muscle stimulation. The subject stands with bended knees and hips on a rocking platform with a sagittal axle, which thrusts alternatively the right and left leg 7-14 mm upwards with a frequency of 27 Hz, thereby lengthening the extensor muscles of the lower extremities. The reflexive reaction of the neuromuscular system is a chain of rapid muscle contractions. We conducted a randomized controlled trial, n=34 (age: mean 67y, range 61-85, 11 female), cross-over design, intervention group 2 months training program three times a week (each session 3x2 minutes), performance tests of all participants every two weeks). The first 19 subjects have finished the intervention period. They reached mean performance gains in chair rising of 18%, strikingly different to the constant values of the controls! We interpret the findings as improvements in muscle power by the oscillative muscle stimulation. PMID- 15758530 TI - Status epilepticus in Indian children in a tertiary care center. AB - OBJECTIVE: To study the clinical profile, immediate outcome and possible risk factors of SE in pediatric age group admitted to pediatric intensive care unit (PICU) in a tertiary care center. METHODS: A retrospective study of case records of 451 neuroemergency patients admitted in PICU in a tertiary care center between January 1993 to April 2000, out of which 30 patients had status epilepticus. They were evaluated for their clinical presentation, laboratory parameters, treatment profile and immediate outcome. RESULTS: The age group varied from 1 to 120 months with mean of 56.6+/- 46.5 months. Seventeen patients were less than 60 months. Sixteen patients (53.3%) presented with SE as first presentation without prior history of seizure activity. Nine patients died (30%) during hospital course. Seizure duration> 45 minutes (p-0.001) and presence of septic shock (p-0.001) were associated with significantly more mortality. CONCLUSION: There is a need to abort seizure activity at the earliest and this improves immediate outcome. PMID- 15758531 TI - Comparative study of complex spina bifida and split cord malformation. AB - OBJECTIVE: To see the difference in clinical profiles, radiological findings and surgical outcome of the group 1 split cord malformation and meningomyelocele (SCM with MMC) from group 2 (SCM without MMC). METHODS: 46 patients of SCM were selected from a total of 138 cases of spinal dysraphism. They were divided into two groups, based on presence or absence of MMC. Group I (SCM with MMC) n =19 patients and Group II (SCM without MMC) n=27 patients. A detail clinical evaluation and MR screening of whole spine of all cases was performed. All patients underwent surgical detethering of cord. After an average follow-up of 1.7 years, the operative results were clinically assessed and statistical significance was calculated. RESULTS: Male to female ratio was 1:09. Mean age of presentation was 3.6 years. Cutaneous markers like tuft of hair, cutaneous haemangioma, etc, had a higher incidence in group II in comparison to group I (50% vs 10.5%). The incidence of motor deficits was significant in group I in comparison to group II (63% vs 40%). The incidences of sensory loss, trophic ulcers, sphincteric dysfunction and muscle atrophy were relatively more common in group I patients, while neuro-orthopedic deformities such as congenital telepes equinovarus (CTEV), scoliosis and limb shortening were more frequent (67%) in group II children as compared to group I (53%). Type I SCM has higher incidence in group I children. Low lying conus were found in 47% patient of group I, while in group II it was noticed in 69%. The associated cranial anomalies like hydrocephalus, ACM and syrinx, were slightly higher in group I patients. At surgery, dysgenetic nerve roots, neural placode, arachnoid bands and atrophic cord were seen mainly in group I. Postoperative complications like, CSF leak, pseudomeningocele and meningitis were more commonly encountered in group I patients. The patients of group II showed better operative outcome compared to group I cases. CONCLUSION: Incidence of SCM with MMC amount to 41% of total SCM cases. Progressive neurological deficit was higher in this group (SCM with MMC) in comparison to the group harboring SCM without MMC. In view of a significant association of SCM in MMC cases, associated with other craniospinal anomalies, a thorough screening of neuraxis (by MRI) is recommended to treat all treatable anomalies simultaneously for desired outcome. PMID- 15758532 TI - Patterns of prescription and drug dispensing. AB - OBJECTIVE: To analyze the patterns of prescriptions and drug dispensing using World Health Organization core drug use indicators and some additional indices. METHODS: Data were collected prospectively by scrutinizing the prescriptions written by pediatric resident doctors and by interviewing parents of 500 outpatient children. RESULTS: The average number of drugs per encounter was 2.9 and 73.4% drugs were prescribed by generic name. Majority of drugs prescribed were in the form of syrups (60.8%). Use of antibiotics (39.6% of encounters) was frequent, but injection use (0.2% of encounters) was very low. A high number of drugs prescribed (90.3%) conformed to a model list of essential drugs and were dispensed (76.9%) by the hospital pharmacy. Certain drugs (5.7%) prescribed as syrups were not dispensed, although they were available in tablet form. Most parents (80.8%) knew the correct dosages, but only 18.5% of drugs were adequately labeled. No copy of an essential drugs list was available. The availability of key drugs was 85%. CONCLUSION: Interventions to rectify over prescription of antibiotics and syrup formulations, inadequate labeling of drugs and lack of access to an essential drugs list are necessary to further improve rational drug use in our facility. PMID- 15758533 TI - Effect of clofibrate in jaundiced term newborns. AB - OBJECTIVE: Clofibrate is a glucuronosyl transferase inducer that has been proposed to increase the elimination of bilirubin in neonates with hyperbilirubinemia. The aim of this study was to characterize the therapeutic effect of clofibrate in neonates born at full term and present with non-hemolytic jaundice. METHODS: A clinical controlled study was performed in two groups of healthy full term neonates. Thirty neonates were treated with a single oral dose of clofibrate (100 mg/kg) plus phototherapy (clofibrate-treated group) while another 30 neonates (control group) received only phototherapy. RESULT: The mean plasma total bilirubin levels of 12th, 24th and 48th hours were significantly lower in the clofibrate-treated group as compared with the control group (P < 0.0001, P < 0.0001 and P = 0.004, respectively). Treatment with clofibrate also resulted in a shorter duration of jaundice and a decreased use of phototherapy (P < 0.0001). No side effects were observed. CONCLUSION: Although other pharmacological agents such as metalloporphyrins and Sn-mesoporphyrin also seem to be effective in decreasing bilirubin production, these products are not available for routine use and cannot be used because the safety of these drugs has to be confirmed prior to their widespread use. Therefore, clofibrate is now the only available pharmacological treatment of neonatal jaundice. PMID- 15758534 TI - Infant feeding - an evaluation of text and taught. AB - OBJECTIVE: Study was conducted to 1) assess knowledge of doctors, medical interns and nurses/ANMs regarding exclusive breast-feeding, management of common problems related to breast feeding and appropriate complementary foods 2) review the above aspects in books commonly read by medical and nursing students. METHODS: Cross sectional study conducted in the department of Community Medicine, Maulana Azad Medical College, New Delhi during a seven-month period on 93 interns, 58 medical officers and 44 nurses/ Auxillary Nurse Midwives (ANMs) with the help of a pre tested structured multiple choice questionnaire and review of some commonly read books of Pediatrics, Community Medicine and Nursing. RESULTS: The concept of 'exclusive breast-feeding' was clear to most health personnel but the fact that water should also not be given was not clear in the books reviewed. The books also lacked emphasis on management of common lactation problems and this was also seen in the knowledge of the health personnel. CONCLUSION: As inappropriate feeding practices are widely prevalent, knowledge of large proportion of health personnel is incorrect and commonly read books are inadequate on this issue, there is need for greater emphasis on this in books and training sessions. PMID- 15758535 TI - Correlates of quality of life with epilepsy. AB - OBJECTIVE: To examine the quality of life of children with epilepsy and to identify the demographic, disease related, and behavioral and emotional functioning variables in the prediction of quality of life of children with epilepsy. METHOD: Forty three children aged 4 to 15 years (Mean=10.3 years) with epilepsy were recruited from the outpatient services of the Department of Pediatrics, of a tertiary care teaching hospital in North India. Quality of life was measured by Impact of Epilepsy Schedule, a 39 items parent reported questionnaire and child's emotional and behavioral functioning at home was assessed by the Childhood Psychopathology Measurement Schedule. RESULTS: Majority of the parents expressed major concerns regarding seizures, treatment by anticonvulsants, present and future problems for the child and problems in parenting. Nearly 40% of the children had psychopathology scores in the clinically significant maladjustment range. Step-wise multiple regression analysis revealed that the psychopathology scores and mother's education accounted for 39% of the variance in the quality of life scores. CONCLUSION: Children with epilepsy have a relatively compromised quality of life and focusing simply on control of seizures may not address the full range of child's emotional and behavioral difficulties. PMID- 15758536 TI - Guest editor: P.S.N. Menon - Editorial: childhood and adolescence growth and growth disorders. PMID- 15758537 TI - Growth hormone therapy. AB - Growth hormone (GH) therapy has revolutionized treatment of children with growth hormone deficiency (GHD). Improved height outcome with final height in the target height range has been achieved in these children. Identification of Creutzfeldt Jakob disease, a deadly prion mediated disorder, in recipients of pituitary GH accelerated the transition from pituitary derived GH to recombinant GH. Once daily subcutaneous administration of the freeze-dried preparation at evening is the recommended mode of GH therapy. Studies have led to use of higher dose of GH for improving height outcome (0.33 mg/kg/week or 0.14 IU/kg/day) albeit at a significantly high cost. Growth velocity increases from 3-4 cm/year before therapy to 10-12 cm/year during the first two years of therapy and is maintained at 7-8 cm/year after a period of two years. Close follow-up with regular clinical and laboratory monitoring is essential for achieving a desirable height outcome. A theoretical unlimited supply has led to wide spread use of GH in a variety of disorders other than GHD. Initially started in children with Turner syndrome, GH has now been used in chronic renal failure, idiopathic short stature and intrauterine growth restriction besides a wide array of newly emerging indications. PMID- 15758538 TI - Molecular defects in the growth hormone-IGF axis. AB - Many previously uncharacterized cases of poor growth in children, i.e. idiopathic short stature, are now attributable to genetic defects in the GH-IGF-1 axis. This paper will provide an overview of the clinical findings of patients identified with various genetic defects of the GH/IGF-1 axis with an emphasis on the more recently described syndromes. PMID- 15758539 TI - Short stature and growth hormone. AB - Normal growth and development is a prime concern during childhood. Accurate assessment is essential for differentiating between normal and abnormal growth. Increased accessibility to growth hormone has equipped the pediatrician and pediatric endocrinologist to treat and improve growth in many clinical scenarios. At the same time, there is added responsibility to use this tool judiciously. This review summarizes the basics of proper growth assessment, differentiation of normal and abnormal growth causes of and works up of short stature, and delineation of indications for growth hormone treatment. PMID- 15758541 TI - Endoscopic correction of severe laryngomalacia. AB - Tracheostomy for management of severe laryngomalacia is associated with significant morbidity and mortality. Two cases are reported wherein the laryngeal abnormality was corrected by ary-epiglottic fold incision and CO2 laser supraglottoplasty. Stridor and respiratory obstruction were relieved and a long term tracheostomy avoided. Endoscopic correction of laryngomalacia offers significant benefits over conventional treatment with tracheostomy in terms of decreased morbidity and improved quality of life. PMID- 15758540 TI - Growth of children with beta-thalassemia major. AB - Hypertransfusion and regular chelation therapy have allowed improved survival in patients with thalassemia major (TM). Despite medical advances, growth failure and hypogonadism remain significant clinical problems in these patients in adolescence. Disproportionate truncal shortening which is common especially among adolescents with thalassemia, is due to platyspondyly resulting from a combination of factors like hemosiderosis, desferrioxamine toxicity or deficiency of trace elements. Although growth hormone (GH) deficiency and GH neurosecretory dysfunction have been described in TM patients, most short TM patients have normal GH reserve. The low serum IGF-1 and IGFBP-3 concentrations in TM patients despite having normal GH reserve and serum GH binding protein levels suggest that a state of secondary GH insensitivity exists. The pubertal growth spurt may be impaired in TM patients going through spontaneous or induced puberty and may have a negative effect on final adult height. GH therapy in dosages ranging from 0.5 1.0 IU/kg/wk has resulted in a significant improvement in growth velocity in short TM children without any adverse effects on skeletal maturation, blood pressure, glucose tolerance and serum lipids. There is limited evidence that GH treatment can result in an improved final adult height in short TM children. Careful and regular clinical and biochemical monitoring should be preformed on these patients while they are treated with GH. PMID- 15758542 TI - Bilateral adrenal abscess in a neonate. AB - A 23-day-old male baby with a history of perinatal hypoxia presented with refusal of feeds and abdominal distension. The child had a right-sided cystic upper abdominal mass and features of neonatal septicemia. Abdominal ultrasound (US) and contrast-enhanced CT scan showed bilateral adrenal abscesses. Laparotomy with drainage of the abscesses successfully treated the condition. The literature on the subject is reviewed. PMID- 15758543 TI - Button battery ingestion. AB - Button batteries represent a special category of pediatric ingested foreign body because of the possibility of serious complications particularly if impacted in the esophagus. We report a case of a 3-year-old girl with severe mid esophageal burns due to a lodged battery. More awareness is required amongst physicians to avert such dangers and ensure prompt removal. PMID- 15758544 TI - Abdominal tuberculosis with autoimmune hemolytic anemia. AB - An eight-year-old male child presented with progressive distension of abdomen, fever, pallor and jaundice with a history of tubercular contact. Investigations were suggestive of abdominal tuberculosis with autoimmune hemolytic anemia. The child responded well to a course of oral steroids with antitubercular treatment. A literature search did not reveal any previous case report of an association between tuberculosis and autoimmune hemolytic anemia. PMID- 15758545 TI - Virchows node: rare presentation of childhood hepatocellular carcinoma. AB - Cervical (supra-clavicular) lymphadenopathy may not always be due to tuberculosis in children. Hepatocellular carcinoma in children even may present as supra clavicular lymphadenopathy (Virchow's node). PMID- 15758546 TI - Megalencephalic leukoencephalopathy with subcortical cysts. PMID- 15758547 TI - Familial chylomicronemia syndrome. AB - Familial chylomicronemia syndrome is a group of rare genetic disorders characterized by deficient activity of an enzyme lipoprotein lipase or apo protein C-II deficiency. Incidence is 1 out of 1,000,000. Alternative names to this syndrome are Type I hyper lipoproteinemia and familial lipoprotein lipase deficiency. PMID- 15758549 TI - Balloon dilatation of pulmonary valve in Tetralogy of Fallot's. AB - In some children of Tetralogy of Fallot's (TOF) presenting with progressive cyanosis, are palliative Blalock-Taussing (BT) shunt may be required. There are no reports of this modality of management in India, though this has been practiced in the other countries. The author reports an infant with Tetralogy of Fallot's who successfully underwent ballon dilatation of the pulmonary valve. Review of literature shows 332 patients with TOF undergoing pulmonary valve balloon dilatation as an alternative to BT shunt in 12 studies with significant increase in pulmonary artery 'Z' score and low incidence of conversion to shunt. This modality of management should be considered in selected patients to change a palliative surgery to an intervention. PMID- 15758548 TI - Langer-giedion syndrome with renal cyst. AB - Here it is reported a 4-year-old boy with Langer-Giedion syndrome (Trichorhino phalangeal syndrome-II), who had characteristic features of TRP II, associated with multiple renal cysts hitherto unreported. This could be a new association in this syndrome that may serve to support the concept of contiguous gene syndrome in patients with TRP II. PMID- 15758550 TI - True tail in a neonate. AB - Distinction between true and pseudo trail in lumbo-sacral region is important since treatment and prognosis are different. Fewer than 40 cases have been reported in literature. The authors report a case of true tail in a neonate, a rare event. PMID- 15758551 TI - Congenital diaphragmatic eventration associated with massive hiatal hernia. AB - Congenital eventration of the diaphragm is uncommon. Its association with a hiatal hernia has not been reported earlier. We report a case of such an association in a 2-month-old boy who presented with tachypnea aggravated by feeding and recurrent vomiting. Diagnostic uncertainty and a unique course of postoperative recovery complicated by gastric volvulus are described. The patient underwent surgical plication of the right diaphragm, followed by an emergency hiatal repair on the first postoperative day. Timely recognition, and repair of the hiatal hernia averted a fatal complication. PMID- 15758552 TI - Adrenomedullin and its related peptide. PMID- 15758553 TI - The genetic contribution of the natriuretic peptide system to cardiovascular diseases. AB - Three types of natriuretic peptides (NP) have been isolated: atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). The NP family elicits a number of vascular, renal and endocrine effects that help to maintain blood pressure and extracellular fluid volume. These effects are mediated by the specific binding of NP to cell surface receptors that have been characterized, purified and cloned from cells of the vasculature, kidney, adrenal gland and brain. There are 3 subtypes of NP receptors: type A natriuretic peptide receptor (NPRA), type B natriuretic peptide receptor (NPRB), and type C natriuretic peptide receptor (NPRC). All 3 subtypes affect cellular second messenger activity. NPRA and NPRB are guanylyl cyclase receptors, and their activation increases cGMP levels. Activation of NPRC results in inhibition of adenylyl cyclase activity. Human NPRA has a high structural homology with human NPRB, and contains a highly-conserved guanylyl cyclase domain. ANP and BNP bind primarily to NPRA, which is found in the vasculature, causing vasodilation and inhibition of vascular smooth muscle cell proliferation. The present paper contains a review of NPs and their receptors and the genetic contribution of the NP system to cardiovascular diseases such as essential hypertension and myocardial infarction. PMID- 15758554 TI - Plasma concentrations of adrenomedullin and ghrelin in hemodialysis patients with sustained and episodic hypotension. AB - Sustained and/or episodic hypotension during hemodialysis (HD) is an important clinical issue. Plasma adrenomedullin (AM) is increased in HD patients with sustained hypotension, but little is known about its implications for episodic hypotension. Ghrelin may also contribute to the pathophysiology of hypotension in HD patients. We evaluated plasma levels of AM and total ghrelin in sustained hypotensive (SH; n = 23), episodic hypotensive (EH; n = 30) and normotensive (NT; n = 23) HD patients. In the EH group, the relationship between low blood pressure during HD and circulating levels of AM and ghrelin was also evaluated. Plasma levels of AM were significantly higher in SH (34.3 +/- 8.3 fmol/ml, p<0.01) than in NT patients (27.6 +/- 5.2 fmol/ml), but not in EH patients (30.8 +/- 6.1 fmol/ml). There was no significant difference of plasma total ghrelin in SH (548.1 +/- 426.5 fmol/ml) and in EH patients (544.6 +/- 174.3 fmol/ml), compared with NT patients (400.0 +/- 219.7 fmol/ml). On the other hand, in EH patients, the "suppressed blood pressure ratio" during HD significantly correlated with plasma AM (r = 0.77, p<0.001) and with total ghrelin levels (r = 0.44, p<0.05). Our results suggest that ghrelin, as well as AM, may play an important role as vasodilator local hormones and regulation of blood pressure during HD, especially the occurrence of EH. Further studies are necessary to clarify the implication of these hormones in the control of hypotension during HD. PMID- 15758555 TI - Rapid differential diagnosis of Graves' disease and painless thyroiditis using total T3/T4 ratio, TSH, and total alkaline phosphatase activity. AB - When thyrotoxic patients are first seen in an outpatient clinic, it is important to make a differential diagnosis of Graves' disease (GD) and painless thyroiditis (PT). Using the three parameters of total T3/T4 ratio, TSH and T-ALP activity, all of which can be obtained within one hour in our hospital, 173 untreated patients with thyrotoxicosis were evaluated for the ratio of each parameter. For GD vs. PT, total T3/T4 (ng/microg) ratio (>20), TSH (<0.005 microU/ml) and elevated T-ALP had a likelihood ratio of 2.14, 2.12 and 4.07, respectively. When a patient had all three positive parameters, the likelihood ratio increased to 5.81, which showed effective synergy. These data suggest that, in addition to total T3/T4 ratio and TSH value, T-ALP activity is a useful parameter for the rapid differentiation of GD. The lower T-ALP activity seen in PT is probably due to the fact that patients develop less severe thyrotoxicosis or visit hospital earlier than patients with GD. Therefore, the diagnostic accuracy for Graves' disease might be increased by using the three parameters in combination. PMID- 15758556 TI - Studies of very severe short stature with severe GH deficiency: from the data registered with the foundation for growth science. AB - The ratio, clinical characteristics, and therapeutic efficacy of hGH treatment in patients with severe short stature (HtSDS below -4SD) with severe GHD (all peak GH values to provocation tests: below 2 mug/L) were studied. From March 1986 to January 1998, 23,110 patients with idiopathic GH deficiency (IGHD) were registered with the Foundation for Growth Science, Japan. These subjects were divided into 5 groups as follows: Group 1 (G1), all subjects; Group 2 (G2), at least one GH peak to provocative test > or = 5 microg/L; Group 3 (G3), 2 microg/L < or = GH peak<5 microg/L; Group 4 (G4), all GH peaks<2 microg/L and HtSDS>-4; Group 5 (G5), all GH peaks<2 microg/L and HtSDS< or = -4. The ratio of G5 was 139 patients (0.6%) out of 23,110 patients with IGHD. In G5, there were no significant differences in birth weight, birth length, gestational age and parental height between G2, G3 and G4. However, asphyxia at delivery was more frequent in G5 and G4 than G2 and G3. Chronological age (CA), bone age (BA) and BA/CA ratio at registration were significantly lower in G5 than G2, G3 and G4. Further, the IGF-I SD score in G5 was significantly lower than those in G2 and G3. After hGH treatment, the final height and final height SDS in G5 remained the lowest, while the DeltaHtSDS value in G5 was the greatest among G2 to G5 groups. In conclusion, the ratio of severe short stature with severe GH deficiency (G5) is only 0.6% of all IGHD cases. Growth failure in G5 seems to occur after birth, and its etiology in G5 seems to be different from that of patients with other forms of IGHD. Early diagnosis and hGH treatment are needed to attain better final height. PMID- 15758557 TI - Differences in TSH receptor binding and thyroid-stimulating properties between TSH and Graves' IgG. Slowly-acting TSH receptor antibody moieties in Graves' sera affect assay data. AB - We analyzed TSH receptor (TSHR) effects, both binding and thyroid-stimulation, of TSH and Graves' IgG. A new TRAb assay system utilizes rhTSHR coated tubes and is comprised of two step incubation, the first incubation with patient serum followed by a second incubation with 125I-bTSH. We called TRAb measured by this method as hTRAb. 125I-bTSH binding capacity of the tube was found close to saturation at 1 hr with 200 microl of 125I-bTSH. Up to 5 hr of first incubation for hTRAb assay revealed significant increases in all hTRAb activities. hTRAb was not affected by second incubation time or dose of 125I-bTSH. When 1 step incubation with 125I-bTSH and Graves' serum was performed, hTRAb again increased significantly with time. A simple competitive equilibrium model could not be applied to these ligands. Second, Graves' IgG and bTSH were compared for in vitro thyroid-stimulation sequentially up to 24 hr, measuring cAMP generation from cultured porcine thyrocytes. While bTSH yielded peak cAMP generation by 8 hr, TSAb revealed more cAMP generation by 24 hr than at 8 hr. We concluded that individual Graves' sera contain heterogeneous TRAb of variable avidities, and that slow-acting TRAb, which may lack biological activity, can be detected by prolonged incubation. PMID- 15758558 TI - A case of hemangiosarcoma in thyroid with severe anemia due to bone marrow metastasis. AB - A 56-year-old woman presented with rapidly enlarging thyroid mass and deep anemia. There was no history of gastrointestinal bleeding, and endoscopic examinations of the gastrointestinal system were normal. Fine needle aspiration cytology from the thyroid nodule was suspicious. After blood transfusion, total thyroidectomy was performed. Postoperative histopathological examination of the specimen revealed hemangiosarcoma of the thyroid. After establishment of the diagnosis chemotherapy was started. But hemoglobin values decreased again and hepatosplenomegaly developed at the second month of surgery. Bone marrow aspiration cytology which was performed demonstrated the same tumoral cells infiltrating bone marrow. The patient died at 12th week after surgery. Thyroid hemangiosarcoma can metastasize to the bone marrow and anemia may be an indicator of the advanced disease. In the differential diagnosis of the anemia, bone marrow metastasis and bone marrow biopsy should be considered in suspected cases. PMID- 15758559 TI - Immeasurably low and non-TRH-stimulatable TSH associated with normal I-123 uptake in two goitrous euthyroid patients: possible existence of other thyroid-hormone regulated thyroid stimulators other than TSH. AB - We described two euthyroid patients with normally functioning goiters, but with persistently undetectable and non-stimulatable TSH levels. Subject 1 was a 64 year-old woman with a large diffuse goiter who has been clinically and biochemically euthyroid without any medication for at least 19 years. Subject 2 was a 31-year-old woman with a small diffuse goiter who has been euthyroid for 4 years. Both patients had persistently undetectable levels of serum TSH, TSH receptor antibodies (TRAb) and thyroid stimulating antibodies (TSAb). Their basal TSH levels were very low and their T3 responses to TRH were very diminished or absent. In contrast, the basal levels of the other pituitary hormones and their responses to LHRH, GRH and CRH stimulation were all within normal limits in both patients. MRI images of pituitary glands, 123I thyroid uptake, and thyroid scans were normal. Ectopic thyroids were not detected on (99m)TcO4- and 123I total body scans. Factors interfering with the measurement of TSH were excluded by recovery studies. In subject 1 a T3-suppression test was positive and a perchlorate discharge test was negative. In subject 2 a T3-suppression test was negative. Euthyroid Graves' disease, subclinical hyperthyroidism, destructive thyroiditis, thyrotoxicosis of extrathyroid origin, central hypothyroidism, and nonthyroidal illness were all ruled out by these observations. These results suggest that an unknown factor, such as thyrostimulin, but not TSH or TSAb, stimulates the thyroid and maintains euthyroidism, and may have a role in the regulation of the hypothalamus-pituitary-thyroid axis. PMID- 15758560 TI - Acute hyperglycemia and activation of the beta-adrenergic system do not exhibit synergistic inhibitory actions on thyrotropin-releasing hormone (TRH)-induced thyroid stimulating hormone (TSH) secretion. AB - Thyrotropin-releasing hormone (TRH)-stimulated thyroid stimulating hormone (TSH) response in normal subjects is suppressed by oral glucose administration. Pharmacologic studies indicate that this suppressive action of glucose is mediated by an increase in hypothalamic somatostatin (SRIH) tone. Since activation of the beta-adrenergic system also suppresses basal TSH secretion by enhancing SRIH release we sought to determine whether isoproterenol alters the suppression of TRH-induced TSH response induced by the stimulation of glucose. Four tests were performed in seven healthy young men: Test 1: 200 microg TRH (iv) at 0 min; Test 2: 100 g oral glucose at -30 min and TRH at 0 min; Test 3: TRH at 0 min with isoproterenol (0.012 microg/kg, iv) infused continuously; Test 4: oral glucose at -30 min, TRH at 0 min with isoproterenol infused continuously. Pretreatment with glucose significantly suppressed TRH-induced TSH secretion. Isoproterenol infusion also suppressed the TRH-induced TSH secretion, but it did not enhance the inhibitory action of glucose on TSH secretion. The degree of suppression induced by glucose was significantly higher than that achieved by isoproterenol. These data suggest that combined administration of glucose and isoproterenol does not exhibit synergistic inhibitory actions on TRH-stimulated TSH secretion, and that the glucose-TRH test could be used for the evaluation of the hypothalamic somatostatinergic activity. PMID- 15758561 TI - A novel spermatogenesis related factor-2 (SRF-2) gene expression affected by TCDD treatment. AB - We have cloned a gene which is specifically expressed at the stage of sexual maturation in the rat testis by means of differential display, and have named it spermatogenesis-related factor-2 (SRF-2). Testicular expression was first detected at 5 weeks of age, and its level of the expression increased up to 7 weeks, and was maintained even at 63 weeks. Its cDNA was 2,789 bp in length and encoded an open reading frame of 718 amino acids. This gene was mainly expressed in the spermatocyte, judging from the result of in situ hybridization. The hypothetical gene product had a motif highly homologous with RabGAP/TBC protein. Taken together, this gene is considered to have some important functions for meiosis. The gene expression was significantly decreased by treatment with TCDD, a candidate endocrine disruptor, when administered to male rats of the nursling period. Body weight and testis weight were decreased by the treatment, but even then the sperm concentration in cauda epididymis was not changed significantly. SRF-2 gene may be a promising biomarker to construct a detection system of uncertain endocrine disruptors. PMID- 15758562 TI - Association of micropenis with Pro185Ala polymorphism of the gene for aryl hydrocarbon receptor repressor involved in dioxin signaling. AB - The prevalence of undermasculinized external genitalia has increased in several countries including Japan, and this phenomenon has primarily been ascribed to the deleterious effects of environmental endocrine disruptors such as dioxins. To examine a possible role of the genetic susceptibility to dioxins in the development of micropenis (MP), we studied the Arg554Lys polymorphism of the gene for aryl hydrocarbon receptor (AHR) and the Pro185Ala polymorphism of the gene for aryl hydrocarbon receptor repressor (AHRR), in 73 boys with MP (34 boys with mild MP from -2.1 to -2.5 SD and 39 boys with severe MP below -2.5 SD) and 80 control males (50 boys and 30 fertile adult males). The allele and genotype frequencies of the AHR polymorphism were comparable between the two groups of males, but those of the AHRR polymorphism were significantly different, with the Pro allele and the Pro/Pro genotype being more frequent in boys with MP than in control males (P-value: 0.0029 for the allele frequency and 0.011 for the genotype frequency). In addition, both polymorphisms were comparable in the allele and genotype frequencies between boys with mild MP and those with severe MP and between control boys and control fertile adult males. The results suggest that the AHRR Pro185Ala polymorphism may constitute a susceptibility locus for the development of MP in response to dioxins. PMID- 15758563 TI - Subclinical hypothyroidism may be associated with elevated high-sensitive c reactive protein (low grade inflammation) and fasting hyperinsulinemia. AB - The association between coronary heart disease and subclinical hypothyroidism (SCH) is unclear. We aimed to determine hs-CRP concentrations in patients with SCH. Seventy-seven patients (age 34.6 +/- 13.7 yr) with SCH (TSH > 4.2 microIU/ml and serum free thyroxine level between 0.932-1.71 ng/dL), and 80 control subjects (age 33.9 +/- 13.3 yr) were studied. Thyroid hormones, C-reactive protein, insulin, glucose, total, HDL, LDL and VLDL-cholesterol levels and HOMA-IR index were also determined. TSH levels of SCH group were higher than control (7.4 +/- 2.9 and 1.55 +/- 0.78 microIU/ml, respectively, p = 0.0001). However, FT4 levels were lower than control subjects (1.18 +/- 0.22 ng/dL and 1.38 +/- 0.26, respectively, p = 0.001). Serum hs-CRP levels of subjects with SCH were higher than control subjects (4.2 +/- 0.8 mg/l and 1.05 +/- 0.3 mg/l respectively, p = 0.0001). Insulin levels of SCH group were higher than control (8.5 +/- 4.3 microU/ml and 7.1 +/- 3.1 microU/ml respectively, p<0.02) but, Homa-IR levels of the two groups were not different. Mean total and LDL-cholesterol levels of SCH group were higher than control (p = 0.01 and p<0.02). We also found a positive correlation between hs-CRP levels and insulin (r = 0.362, p = 0.002 in men, r = 0.358, p = 0.0001 in women), TSH (r = 0.611, p = 0.0001 in men, r = 0.411 p = 0.0001 in women), and prolactin (r = 0.340, p = 0.01 in men r = 0.553, p = 0.0001 in women). CONCLUSIONS: Patients with SCH, irrespective of gender, have higher serum hs-CRP, insulin, total and LDL-cholesterol levels than healthy subjects. 2- High hs-CRP level, and thereby low grade inflammation may be associated with fasting hyperinsulinemia before insulin resistance becomes evident in patients with SCH. PMID- 15758564 TI - Increased risk for atherosclerosis estimated by pulse wave velocity in hypothyroidism and its reversal with appropriate thyroxine treatment. AB - Pulse wave velocity (PWV) is known to represent arterial stiffness and is established as a marker for cardiovascular risk and a prognostic factor for mortality in the case of chronic renal failure or hypertension. The application of an automated apparatus for measuring brachial-ankle pulse wave velocity (baPWV) has made PWV measurement non-invasive, easier to screen for cardiovascular risk and as a result, baPWV measurements have become widely applied in clinical practice in recent years. We assessed the baPWV in 7 flank hypothyroidism patients and 28 subclinical hypothyroidism patients. In comparison with age matched healthy controls, 3 hypothyroid patients had advanced values and by replacement therapy, all 7 subjects showed improvement in their baPWV values (1531.2 +/- 242.7 to 1330.2 +/- 208.6 cm/s, p<0.05). In 28 subclinical hypothyroid subjects, 71% also had accelerated baPWV values for their age. Ten subjects (36% of all) had neither hypertension, hyperlipidemia, diabetes nor were taking any medication, and yet 8 patients out of 10 showed advanced baPWV values compared to age matched mean values. The baPWV was not correlated to TSH or total cholesterol levels, and was associated with only age and blood pressure (p = 0.01, <0.001, respectively), which are widely demonstrated as the characteristics for baPWV. In two subclinical hypothyroid subjects, who were normotensive and had no dyslipidemia, thyroxine treatment was performed and the baPWV decreased with unchanged blood pressure and total cholesterol levels. We concluded that the arterial wall stiffness tends to be increased in both overt and subclinical hypothyroid patients, and an appropriate treatment could reverse the abnormalities. It is possible that the initiation of adequate treatment in subclinical hypothyroidism may reduce the cardiovascular risk. PMID- 15758565 TI - Dexamethasone infusion testing in the diagnosis of Cushing's syndrome. AB - Cushing's syndrome and its various aetiologies is a markedly difficult diagnosis to make given its subtle signs, sometime cyclical nature, and the lack of a single definitive diagnostic test. Although a great variety of diagnostic tests have been developed to assist in the diagnosis, even with the best clinical acumen, biochemistry and medical imaging the diagnosis can remain elusive. The long low and high dose oral dexamethasone suppression test is cumbersome, costly and often requiring an extended inpatient stay. The utility of the dexamethasone suppression test would be greatly enhanced if it could be performed as a short outpatient procedure. In this study we sought to confirm and refine the clinical utility of the high dose 4 mg intravenous dexamethasone suppression test as an alternative diagnostic test for Cushing's syndrome. There were a total of 31 subjects: 8 patients with proven pituitary Cushing's disease, 3 with primary adrenal tumors, 10 with pseudo-Cushing's syndrome and 10 healthy controls. All subjects with pseudo-Cushing's syndrome suppress serum cortisol at +5 and at +24 hours. In subjects with pituitary Cushing's disease, 7 out of 8 (88%) had serum cortisol suppressed at +5 hours but rebounded at +24 hours to at least 70% of the original serum level. Primary adrenal tumors showed a pattern of non-suppression throughout. The 4 mg intravenous dexamethasone suppression test is excellent in ruling out pseudo-Cushing's syndrome. This test is much simpler and more convenient than the oral dexamethasone suppression test in confirming clinical suspicion of pituitary Cushing's disease. PMID- 15758566 TI - 3beta-Hydroxysteroid dehydrogenase in human aorta. AB - 3beta-Hydroxysteroid dehydrogenase (3beta-HSD) is known to be involved in steroid production and/or metabolism and to be expressed in many tissues including adrenal cortex. Expression of this enzyme has also been elucidated in human cardiovascular system but its details remain largely unknown. Therefore, in this study, we examined the status of 3beta-HSD in postmortem human aorta utilizing RT PCR and immunohistochemical analysis. Both mRNAs and immunoreactivity for the enzyme were detected predominantly in female aorta, and in those with mild atherosclerotic changes. In addition, immunohistochemical study demonstrated that immunoreactivity for 3beta-HSD was detected in vascular smooth muscle cells (VSMCs) of aorta. These findings all indicated that steroidogenesis via 3beta-HSD may occur in VSMCs of human aorta, possibly related to gender differences and/or the degree of atherosclerosis. PMID- 15758567 TI - Spontaneous remission of functioning pituitary adenomas without hypopituitarism following infarctive apoplexy: two case reports. AB - Functioning pituitary adenomas may exhibit spontaneous remission after pituitary apoplexy usually in association with hypopituitarism. We report two patients who presented with sudden headache and double vision, showed a ring-enhanced sellar tumor on MRI, underwent transsphenoidal surgery that revealed a coagulation necrotic adenoma without massive hemorrhage, and showed normal pituitary function after the surgery. Definitive diagnoses were made based on immunohistochemistry of the necrotic cells. The findings were consistent with the presence of selective infarct of a GH adenoma and a prolactinoma that had led to remission of acromegaly and menstrual disturbance, respectively, without pituitary insufficiency. In contrast to hemorrhagic apoplexy, infarctive apoplexy tends to affect only the tumor and thus presents with mild symptoms and lack pituitary deficiencies. PMID- 15758568 TI - Slow postoperative decline in blood concentration of insulin-like growth factor-1 (IGF-1) in acromegalic patients. AB - Recent criteria for cure of acromegaly require normalization of the insulin-like growth factor (IGF-1) level. A retrospective study was conducted to assess postoperative sequential changes in the blood IGF-1 level and to determine the appropriate timing for endocrinologic assessment of the effect of surgery. Blood IGF-1 levels were measured at least 3 times (4.9 +/- 2.0, mean +/- SD) during the first postoperative year in 36 acromegalic patients whose glucose tolerance test results, obtained 3 months after surgery, showed the growth hormone level to be suppressed to under 1 ng/mL. Postoperative IGF-1 parameters, i.e. the actual IGF level and %IGF-1 compared to the nadir during the first postoperative year, rapidly decreased during the first 2 postoperative weeks and then slowly declined over the next 2 weeks. They reached a plateau (stable nadir) during the 2nd postoperative month. Assessment of the postoperative endocrinologic status should be delayed at least one month after surgery in acromegalic patients. PMID- 15758569 TI - GnRH antagonist-induced down-regulation of the mRNA expression of pituitary receptors: comparisons with GnRH agonist effects. AB - In order to compare the mechanism for the down regulation of the mRNA expression of pituitary receptors induced by GnRH antagonist (GnRHant) to that by GnRH agonist (GnRHa), we examined the effects of GnRHant (Cetrorelix, 333 mug/kg/day), GnRHa (leuprolide depot, 333 microg/kg), and GnRHant combined with GnRHa on LH response to exogenous GnRH, pituitary LH content, LH beta subunit mRNA, and GnRH receptor (GnRH-R) mRNA levels at 2, 5, 24, 72 hours, and 7 days after the treatment in ovariectomized rats. GnRHant significantly decreased serum LH, the LH response of the pituitary to exogenous GnRH, and the pituitary LH content compared to the control treatment, though GnRHa significantly increased serum LH. GnRHant with GnRHa significantly diminished the GnRHa-induced flare-up phenomenon. GnRHant significantly decreased LH beta mRNA and GnRH-R mRNA levels, but the magnitude of the decrease in these mRNA levels by GnRHant was significantly less than those by GnRHa until 72 hours following treatment. Prolonged treatment of GnRHant caused a marked inhibition of LH beta mRNA and GnRH-R mRNA expression, similar to that caused by GnRHa. Combination treatment with GnRHa and GnRHant was demonstrated to decrease LH beta mRNA and GnRH-R mRNA levels as much as GnRHa alone and GnRHant alone over 7 days of the treatment. The present study showed differences between GnRHant and GnRHa treatment in the reduction of GnRH-R mRNA levels up to 72 hours after the treatment, and indicated that the suppression of GnRH-R mRNA by GnRHant was the maximal by GnRHa 7 days after the treatment because more profound suppression was not observed upon additional treatment with GnRHa. The findings in the present study support the hypothesis that the mechanism by which GnRHant leads to down-regulation of the mRNA expression of pituitary receptors is similar to that of GnRHa. PMID- 15758570 TI - Correlation of the dysmetabolic risk factors with different anthropometric measurements. AB - Metabolic syndrome is a common disorder in Taiwan. For this study 431 subjects were randomly selected from visitors to the Department of Health Management. Blood pressure, blood glucose, lipid, uric acid levels and anthropometric measurements with immunoreactive insulin (IRI) and leptin levels were all correlated. We randomly selected 431 subjects who visited the Department of Health Management. Whole body three-dimensional (3-D) laser scanner scans were employed for the anthropometric measurements. The metabolic index (MI) was designed using anthropometric parameters. Of the 431 subjects, 50% had displayed a body mass index (BMI) equal to or exceeding 25 kg/m2. Pearson correlation coefficient and multiple regression analysis revealed that MI constituted another index for correlating metabolic parameters by comparing MI with BMI and waist circumference to hip circumference ratio (WHR). Most data related to metabolic syndrome showed statistically significant differences between high and low IRI groups, comprising uric acid, total cholesterol, fasting plasma glucose, triglyceride, LDL, Chol/HDL ratio, and LDL/HDL ratio. Both IRI and leptin revealed statistical association with BMI, WHR, waist cross section area to hip cross section area ratio (WHAR), and MI in the study. Hypercholesterolemia appeared in 14.6% of the subjects. Elevated low-density lipoprotein (> or = 130 mg/dL) affected 36.9% of the subjects. In conclusion, MI calculated from 3-D body scanner correlated with many important metabolic risk factors and associated with clinical disorders like DM, hyperlipidemia, hyperuricemia and hypertension. PMID- 15758571 TI - Thyrotoxic periodic paralysis in a Turkish male; the recurrence of the attack after radioiodine treatment. AB - Thyrotoxic hypokalemic periodic paralysis (THPP) is a rare complication of hyperthyroidism and an uncommon form of hypokalemic periodic paralysis. Its differentiation of more common forms of periodic paralysis is important because aggressive treatment can place the patient at risk for rebound hyperkalemia. Treatment of the underlying thyroid dysfunction cures the muscle symptoms. Here we describe a 37-year-old Turkish male with THPP whose paralysis attack recurred soon after administration of radioactive iodine. PMID- 15758572 TI - Fos plays no role in apoptosis of epithelia in the mouse male accessory sex organs and uterus. AB - Roles of Fos in apoptosis of epithelia in the mouse male accessory sex organs and uterus were investigated using Fos-deficient mice. Normal 30- and 50-day-old and Fos-deficient 50-day-old male and female mice were castrated, and testosterone propionate and estradiol-17 beta were daily injected into male and female mice, respectively, for 5 days. An apoptotic index (a percentage of apoptotic cells) in the epithelium was examined from the day following the last injection (day 1) to day 8. The body weights and the weights of the ventral prostate (VP), coagulating gland (C), seminal vesicle (SV) and epididymis (Ep) and uterus of 50-day castrated Fos-deficient mice on day 1 suggested that the development of these mice corresponded to that of 30-day-castrated normal mice at the most. The extents of apoptosis estimated by an apoptotic index in the VP, C, SV, Ep and uterus in 50-day-castrated Fos-deficient mice were comparable to those in 30-day castrated normal mice. The extents of apoptosis in the SV, Ep and uterus in 30 day-castrated normal and 50-day-castrated Fos-deficient mice were similar to those in 50-day-castrated normal mice, while the extents of apoptosis in the VP and C in the former two groups of mice were less than those in the latter mice. The present results show that Fos-deficiency does not affect apoptosis in the SV, EP and uterus. However, the extents of apoptosis in the VP and C were less in 50 day-castrated Fos-deficient mice than in 50-day-castrated normal mice. This seems to be due to the retarded development of 50-day-castrated Fos-deficient mice, but not to a role of Fos in apoptosis. PMID- 15758573 TI - Dose of anti-thyroid drugs can effect remission in Grave's disease. PMID- 15758574 TI - [Vaccine for human immunodeficiency virus (HIV)--relevance of these days]. AB - Since 1980 more than 25 million people have died from acquired immunodeficiency syndrome (AIDS), which results from infection with human immunodeficiency virus (HIV). Number of new cases increases very threateningly. One and the most effective method to stop the progress of epidemic is the development of the vaccine for HIV. There is the presentation of the first stage of the vaccine for HIV testing (structure, methodology), which is now on trial in St. Pierre hospital, Brussels University. HIV characteristics which inflame the process of the vaccine development, historical facts and facts about vaccines on trial in these days are reviewed in this article. More than 10,000 volunteers have been participating in various clinical trials since 1987. The development of the vaccine is a very difficult, long-terming (about 8-10 years) and costly process. The process of the vaccine testing is very difficult in developing countries where the infection spreads the most rapidly. Available data confirm that the vaccine must be multi-componential, inducing cellular, humoral immunity against various subtypes of HIV. The vaccine cannot protect fully but the changes of the natural infection course could decrease virulence, distance the stage of AIDS, and retard the spread of the epidemic. PMID- 15758575 TI - Prevalence, clinical features and accompanying signs of post-traumatic headache in children. AB - The aim of the study was to investigate the prevalence and clinical features of headaches and their accompanying signs in children with mild traumatic brain injury, as well as to evaluate their changes over time. MATERIAL AND METHODS: The research involved two groups of 4-16 year-old children: the case group of 301 children who had experienced a single mild traumatic brain injury, and the control group--301 children who had suffered from any other mild body injury without head trauma. Groups were matched according to gender, age, date of admission to hospital. The period between the date of trauma and examination was at least one year (median 7 months). Standardized questionnaires were sent by post to parents from both groups. Parents were asked about character, frequency, dizziness and concomitant symptoms. In total, 102 matched pairs were analyzed. RESULTS: During the year before the filling in the questionnaires 114 (57.3%) parents indicated headaches: 64 (62.7%) in the case, and 50 (49%) in control group. Frequent (> or =8 days per month) headaches prevailed in children with mild traumatic brain injury (p=0.039); however, their prevalence decreased from 43.8% to 12.5% (p=0.01) with increasing time interval between the date of trauma and the inquiry. By the character, duration, intensity of the headaches, none of the features (except dizziness) showed any difference between the groups. Thirty three (51.6%) parents from the case and 16 (32%) from the control group indicated that dizziness accompanied headaches (p=0.036). Forty-five (44.1%) parents from the case group and 28 (27.5%) parents from the control group indicated that dizziness appeared separately from headaches (p=0.013). According to the data for the last month before the inquiry, there was no statistically significant difference between both groups when estimating the dynamics in time. CONCLUSIONS: Headaches are not more prevalent in children with mild traumatic brain injury, compared to children with other mild body injuries. The frequency of headaches, as well as the prevalence of dizziness in children with mild traumatic brain injury decreases with time. PMID- 15758576 TI - [Value of activities and participation of patients with circulation diseases]. AB - The aim of our study was to evaluate the disturbances of activities and participation of inpatients after stroke and myocardium infarction. Activities and participation disorders were evaluated by international classification of functioning, disability and health. Our study material consisted of inpatients after stroke (I group) and inpatients after myocardium infarction (II group). The age average was 70.4+/-7.4 years in the first group and 65.7+/-11.7 years in the second group. Activities and participation were respected by the following scale: 0--no difficulty, 1--mild difficulty, 2--moderate difficulty, 3--severe difficulty, 4--complete difficulty. We determined, that changing and maintaining body position, learning to write, moving around activities were the most confused (3-4); thinking and copying were confused moderately (2) in the first group. Changing and maintaining body position, focusing attention, calculating and handling stress activities were confused moderately (2) in the second group. International classification of functioning, disability and health allows evaluating activity limitations and participation restrictions in blood circulation disorders. We noticed, that basic learning, applying knowledge, general tasks and demands, communication, maintaining a body position, and self care were more confused in patients after stroke. Moving around, focusing attention, and handling stress were more confused in patients after myocardium infarction. It is important to evaluate activity limitation and participation restrictions for determining the extent of rehabilitation. PMID- 15758577 TI - Fine needle aspiration cytology diagnosis of gastrointestinal stromal tumors utilizing scanning electron microscopy. AB - BACKGROUND: Digestive stromal neoplasms are the most frequent undifferentiated mesenchymal tumors. The outcome of these malignancies is difficult to predict and the histogenesis is still controversial. However, the frequent and specific expression of CD117 (c-kit) by these tumors could imply an origin from interstitial cells of Cajal. Our objective was to analyze the role of fine needle aspiration cytology, cell block preparation, and immunocytochemistry in the interpretation of gastrointestinal stromal tumors, and to establish scanning electron microscopy as a useful research aid for pathologic changes of the surface cells of gastrointestinal stromal tumors, not totally appreciated by light microscopy. MATERIAL AND METHODS: Twelve cases of gastrointestinal stromal tumors were included in this study, in which fine needle aspiration cytology was performed. RESULTS: On aspirated material, the tumor cells formed closely packed cohesive tissue fragments with high cellular density often in bloody background, or fascicles with parallel side-by-side arrangements of the nuclei. On cell block biopsy material, gastrointestinal stromal tumors were highly cellular spindle or epithelioid tumors with basophilic appearance. Immunocytochemically, they were CD117 positive in all twelve cases, CD34 positive in nine, weakly smooth muscle actin-positive in five, and S-100 and GFAP-negative in all cases. The scanning electron microscopy study showed a strong correlation with the cytomorphological profile. CONCLUSIONS: Gastrointestinal stromal tumors show a broad morphologic variety, but nuclear pleomorphism by cytology alone, rarely correlates with malignant potential. In the appropriate clinical and radiological setting, a confident diagnosis of gastrointestinal stromal tumors can be documented by fine needle aspiration cytology, cell block, immunocytochemical, and scanning electron microscopy results. PMID- 15758578 TI - [Aberrant promoter methylation of tumor suppressor genes in serum from lung cancer patients: frequency and correlation with clinicopathological characteristics]. AB - OBJECTIVE: The aim of this study was to determine aberrant promoter methylation of tumor suppressor genes in genomic serum DNA from lung cancer patients and to evaluate the association between methylation of genes and clinicopathological characteristics. MATERIALS AND METHODS: Genomic serum DNA from 46 lung cancer patients and 14 healthy control persons was examined. Nested methylation-specific PCR approach was used for detection of methylated genes in serum. RESULTS: Aberrant promoter methylation in serum from lung cancer patients was detected in 2.3% (1/43) for O6-methylguanine-DNA-methyltransferase, in 11.1% (5/45) for p16INK4a, in 41.3% (19/46) for retinoic acid receptor beta, in 4.5% (2/44) for RAS association domain family 1A, in 40.9% (18/44) for fragile histidine triad, in 34.9% (15/45) for p14(ARF), in 6.5% (3/46) for adenomatous polyposis coli 1A, in 78.1% (25/32) for Ecad, in 5.7% (2/35) for adenomatous polyposis coli 1B, in 0% (0/36) for death associated protein kinase. None of the death associated protein kinase, O6-methylguanine-DNA-methyltransferase, p16INK4a, fragile histidine triad or adenomatous polyposis coli 1A promoter regions were positive for methylation in serum DNA from healthy persons. A total of 78.3% of the samples from lung cancer patients had methylation in at least one of the genes tested. The methylation status of fragile histidine triad was associated with that of p14ARF(p=0.021), as was retinoic acid receptor beta--with that of adenomatous polyposis coli 1A (p=0.04). Methylation of adenomatous polyposis coli 1A was detected more frequently in tumor with pleura involvement (p=0.079), retinoic acid receptor beta was observed more frequently in undifferentiated tumor than in better-differentiated tumor (p=0.029). The methylation status of RAS association domain family 1A and fragile histidine triad showed a tendency to be associated with an advanced tumor size. In addition, tumors with an advanced pathological stage showed epigenetic alteration of the RAS association domain family 1A promoter with a higher frequency. The methylation index was also associated with an advanced tumor size (p=0.033). CONCLUSION: Approximately 80% of the samples from lung cancer patients had methylation of the tumor suppressor gene and it might be associated with more aggressive tumor. Detection of epigenetic alterations in serum may provide basis for the early and noninvasive lung cancer diagnosis and customized therapy. PMID- 15758579 TI - Denys-Drash syndrome. AB - Constitutional missense mutations in the WT1 gene are usually associated with Denys-Drash syndrome. This rare syndrome is characterized by a rapid progressive nephropathy, male pseudohermaphroditism, and an increased risk for Wilms tumor. We report on a patient with incomplete Denys-Drash syndrome, which was evident by the clinical data and proved by molecular genetics methods. The patient has the mutation p.R394W in the WT1 gene and clinical symptoms of Denys-Drash syndrome. PMID- 15758580 TI - [Chronic poisoning by metallic lead from locations of gunshot wound]. AB - A patient injured by shooting to head and shoulder with lead pellets was treated. Fragments of lead pellets were lodged in areas of cheek, forehead and neck. After three months symptoms of lead intoxication occurred, however there was no accumulation of lead in hair. During surgical treatment, it was determined by methods of computed tomography and Zeeman atomic absorption spectrometry that particles of metallic lead dissolved mostly in highly vascularized areas of tissues. The main intoxication occurred from the area of venter frontalis where lead particles were mostly deformed. Intoxication did not disappear as long as all fragments of lead pellets were removed surgically. Resorbed lead was partially accumulated in the bone, because amount of lead eliminated with urine was less than that resorbed into the blood. PMID- 15758581 TI - Psychophysical measurements of illusion of the puffy circle. AB - The strength of an illusion of curvature created by an equilateral triangle, square or pentagon inscribed in a circle has been measured in the psychophysical experiments. The arcs of the circle looked as if they were bowed outwards in the stimuli of various sizes, but, at a fixed diameter of the circumscribed circle, the triangles produced the strongest, and the pentagons, the weakest illusion. The strength of the illusion augmented with the stimulus diameter. Concave and convex sides of the inscribed figures caused less illusory effect than the straight ones. Similar distortions of the stimuli have been observed in the output of a neurophysiological model of spatial frequency filtering of images, and the computed curves resembled those of the experimental data, in respect to their shape and quantitative values. PMID- 15758582 TI - Age-related changes in the structure of myocardial collagen network of auricle of the right atrium in healthy persons and ischemic heart disease patients. AB - The objective of the study was to examine and evaluate morphometrically age related changes in the structure of myocardium collagen network of auricle of the right atrium in control group persons, who were not diagnosed with cardiac pathology leading to heart lesion or overload, and in ischemic heart disease patients. Material of 56 persons of both genders aged 20-94 years was used for study purposes. Biopsy material of 17 healthy persons (control group, average age 60.53+/-9.89 years) and autopsy material of 39 ischemic heart disease patients (average age 63.83+/-15.67 years) taken from the basis of auricle of the right atrium (specimen size--2 mm x 2 mm) were examined. Morphometric analysis of collagen network was performed using histologic and video morphometric methods. After this investigation we evaluated quantitative parameters of the bundles of collagen net--namely area, number, perimeter. The percentile occupied area of bundles in control group was 17.6+/-2.5%; ischemic heart disease patients group- 26.8+/-2.9%; number of bundles was 4179+/-1073 and 2523+/-867; perimeter--24163+/ 3308 mm and 23426+/-409 mm, respectively. After investigation of age-related changes of collagen network in control group and ischemic heart disease patients' group, which did not statistically significantly differed by age, we determined that collagen network area in auricle of the right atrium increased with age in both groups, however, spatial distribution of collagen network was different. Collagen network area enlarged with lengthening of its fibers along cardiomyocytes in control group. In ischemic heart disease group, it enlarged both in parallel to cardiomyocytes and by separate collagen fibers merging into bigger bundles. Fibrillar collagen network area and its total perimeter of healthy persons increased with age, and number of fibers did not change. Consequently, collagen fiber area of one location increased with age; its shape, judging by in parallel increasing total perimeter, became branchier, i.e. proliferated in endomysium in parallel with cardiomyocytes. In ischemic heart disease group fibrillar collagen network percentage area increased with age, however, total perimeter and number of separate fibers in visual field decreased. Consequently, in ischemic heart disease group separate collagen fibers merged, their locations enlarged, taking an integral structure, which allowed assuming development of interstitial fibrosis. PMID- 15758583 TI - [Assessment of quality of life in the middle-aged Kaunas population]. AB - The aim of this study was to evaluate validity and reliability of the WHOQOL-100 questionnaire in the middle-aged Kaunas population and to assess quality of life among men and women and age groups. MATERIAL AND METHODS: The random sample consisted of 1347 Kaunas men and women aged 35-64 and stratified by age and sex (response rate 62%). They were examined in 2001-2002 using self-administered of the WHOQOL-100 questionnaire. Approval from the Ethics Committee of Kaunas University of Medicine was obtained and participants signed a written informed consent prior to filling in a questionnaire. The direct method was used for age standardization using European population as a standard. Validity and reliability of the questionnaire were evaluated by Pearson's correlation coefficients and by Cronbach's alfa. RESULTS: The reliability of the questionnaire among items was found to be very high for all domains (Cronbach's alfa from 0.77 to 0.91). The retest reliability was high or very high for all domains (0.64-0.89). Women scored lower in all domains as compared to men, except spirituality (p<0.001). The scores of the questionnaire decreased with age among both men and women. Women aged 45-54 scored lower in the quality of life, physical and independence level domains, whereas men were lower in the social relationship and spirituality domains as compared to the youngest group (p<0.05). CONCLUSION: The WHOQOL-100 questionnaire is a reliable and valid measure of quality of life in the middle aged Kaunas population; older age and female gender are the determinants of worse quality of life. PMID- 15758584 TI - [Trends in morbidity from acute myocardial infarction, risk factors and risk of ischemia]. AB - THE AIM: To assess the linkages of trends between the mean values of main risk factors of ischemic heart disease, the overall high risk of coronary within 10 years and morbidity from acute myocardial infarction in Kaunas population aged 35 64 years during the period from 1983 to 2002. MATERIAL AND METHODS: Data sources were four cross-sectional surveys (1983-1984, 1986-1987, 1992-1993, 2001-2002) and population-based ischemic heart disease register (1983-2001) of Kaunas population aged 35-64 years. Data collection was conducted according to the method using in the Multinational Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA) project. The overall high risk developing a fatal or non-fatal myocardial infarction or sudden death within 10 years was calculated by PROCAM Risk Score. RESULTS: During 1983-2002 the mean value of systolic blood pressure decreased among men and women (respectively, from 138.8 to 136.4 mmHg, p<0.05 and from 139.7 to 133.2 mmHg, p<0.05). Body mass index decreased among men (from 27.5 to 27.4 kg/m2, p<0.05), and diastolic blood pressure decreased among women (from 86.5 to 82.7 mmHg, p<0.05). In 1983-2002 the overall high 10-year risk also decreased in men and women (respectively, from 20.4 % to 16.3% and from 7.9% to 5.4%). During 1992-2001 the trend of morbidity from acute myocardial infarction was declining among men (by - 4.6 %/year, p<0.01) and among women (by- 1.3 %/year, p>0.05). CONCLUSION: Trends of the overall high 10-year coronary risk and morbidity from acute myocardial infarction were analogous in both genders. PMID- 15758585 TI - [Diagnosis and management of chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is an increasing health problem and one of the leading causes of morbidity and mortality worldwide. Cigarette smoking remains the main risk factor. COPD is preventable, readily diagnosable and treatable disease. The appropriate and early use of spirometry for diagnosis is of importance. A comprehensive treatment plan for managing patients with COPD involves the use of pharmacological as well as nonpharmacologic interventions. Smoking cessation can substantially reduce the risk for the development or rate of progression of COPD. Bronchodilator therapy is a basis in symptomatic treatment. Inhaled steroids might reduce frequency and severity of exacerbation and can be effectively combined with long acting beta2 agonists. Pulmonary rehabilitation benefits most patients. Patients with hypoxemia suffering from more severe disease may require a long-term oxygen therapy. Surgical intervention may help a limited number of patients. PMID- 15758586 TI - Quantitative outcome measures of cartilage repair in patients treated by tissue engineering. AB - Reliable and reproducible outcome measures are essential to assess the efficacy of competing and novel tissue-engineering techniques. The aim of this study was to compare traditional histological analyses with newly developed quantitative biochemical outcome measures for the repair of articular cartilage. The production of a new anti-peptide antibody and the development and validation of a novel method for the extraction and immunoassay of type I collagen are described. The assay was used, in conjunction with existing assays for type II collagen and proteoglycans, to measure levels of the matrix components in repair tissue biopsies obtained from patients treated with the new tissue-engineering therapy Hyalograft C. Frozen sections cut from the same biopsies were stained for proteoglycans, using safranin O, and immunohistochemical analysis was used to assess type I and II collagen staining. Although there was general agreement between the extent of staining and the amounts of the three matrix components, there was a large degree of overlap in biochemical content between biopsies classified histologically on the basis of low, moderate, or abundant staining. The results demonstrate that histological grading of matrix protein abundance to classify repair cartilage as hyaline or fibrocartilagenous is often misleading. In addition, we demonstrate for the first time the ability to measure collagen cross-links in repair tissue biopsies and show that it can be used as a surrogate marker for tissue maturity. Our new range of biochemical techniques provides a standardized method to assess the quality of both engineered cartilage produced in vitro and repair tissue biopsies obtained after in vivo implantation. PMID- 15758588 TI - Genetic influences on outcome following acute neurological insults. AB - PURPOSE OF REVIEW: To examine the evidence for a genetic influence on clinical outcome after a variety of acute neurologic events. RECENT FINDINGS: Clinical outcome after brain injury is variable and cannot easily be predicted. It has been proposed that genetic polymorphisms may have an important role in determining outcome from a number of conditions, including acute neurologic events. Apolipoprotein E, an important mediator of cholesterol and lipid transport in the brain, is coded by a polymorphic gene (APOE). The APOE epsilon4 allele has been associated with unfavorable outcome after traumatic brain injury (TBI), hemorrhagic stroke and subarachnoid hemorrhage (SAH). Genes involved in other pathophysiological processes, such as cytokine genes in neuroinflammation, are now being implicated. For example interleukin-6 (IL-6) promoter polymorphisms are a risk factor for poor outcome after ischemic stroke, and may have an effect after traumatic brain injury. The emerging importance of a number of other gene polymorphisms is outlined in the review. SUMMARY: There is evidence demonstrating the epsilon4 allele of APOE predisposes to poor outcome after TBI, hemorrhagic stroke and SAH, but not ischemic stroke. The reason for this difference is unclear but it suggests there may be differences in the key mechanisms underlying the response to different types of insult. The role of other gene polymorphisms is being increasingly explored but there is still a need for larger prospective studies looking at larger panels of gene polymorphisms. PMID- 15758589 TI - Cognitive outcome in traumatic brain injury survivors. AB - PURPOSE OF REVIEW: This review discusses recent studies investigating the cognitive and psychiatric outcome of posttraumatic brain injury. In addition, it aims to highlight key areas for future research. RECENT FINDINGS: Detailed cognitive assessments have revealed particular deficits in processing speed in the visual domain and the detrimental impact of interference on attentional performance. A pilot functional imaging study revealed neural changes in survivors performing a response inhibition task, even when matched to controls on behavioural indices. Recent psychiatric studies highlight the incidence of these disorders in the survivors and attempt to characterize distinct psychiatric profiles. Adult and child survivors appear to show differential difficulties. Successful rehabilitation strategies addressing these psychiatric and cognitive deficits include holistic intensive neuropsychological interventions and the introduction of electronic devices. Systematic randomized trials are needed to provide an adequate evidence base for clinical practice. The potential for cognitive enhancement using psychopharmacological agents has yet to be exploited. These treatments may lead to improved quality of life for traumatic brain injury survivors and their families. SUMMARY: Survivors of head injury show a diverse pattern of cognitive and psychiatric profiles. Recent research highlights the nature of some of these deficits and possible ways to enhance functioning. However, the area is well poised for rapid progress in the understanding of cognitive and emotional dysfunction following traumatic brain injury (TBI) and its rehabilitation through neuropsychological and psychopharmacological means. PMID- 15758590 TI - Refractory status epilepticus. AB - PURPOSE OF REVIEW: Although conventional anticonvulsant agents can terminate status epilepticus in most cases, a substantial minority of patients develops medically refractory status and requires more aggressive care. This review explores the options available. RECENT FINDINGS: Increasing numbers of previously unexpected etiologies for refractory status epilepticus continue to be reported. There are also some promising new therapies on the horizon, both for the short and the longer terms. SUMMARY: Refractory status epilepticus, while a challenge to the intensivist, can be treated with drugs that are commonly used by intensivists. The cooperation of an interested electroencephalographer is vital. PMID- 15758591 TI - To clip or to coil acutely ruptured intracranial aneurysms: update on the debate. AB - PURPOSE OF REVIEW: The purpose of this review is to consider why the International Subarachnoid Aneurysm Trial (ISAT) led to such an uproar, provide a summary of the criticisms of the study and responses from the investigators, and discuss the implications of ISAT. RECENT FINDINGS: The ISAT was destined for controversy. It pitted two different treatment modalities directly against each other: one a highly evolved, established means for repairing ruptured intracranial aneurysms and the other a radically different, developing approach. The criticisms of the ISAT fall into several categories. They include study site selection, patient selection and the definition of clinical equipoise, relative expertise of the surgeons and interventionists, and the duration and type of follow-up. While some of the criticisms of ISAT reflect unavoidable statistical, design and temporal constraints and thus are of little impact, others are more consequential and revolve around the need for clinical equipoise, and how that was defined across centers. In addition, the question of surgical expertise needs to be addressed. Finally, long-term outcome data are needed. SUMMARY: The study was well designed and executed. The published results met the most up-to-date standards for reporting clinical trials. Important questions remain to be answered, including how to define clinical equipoise and the long-term durability of coiling aneurysms. PMID- 15758592 TI - Critical illness myopathy and neuropathy. AB - PURPOSE OF REVIEW: To present the major pathophysiological and diagnostic features of critical illness myopathy (CIM) and polyneuropathy (CIP), and to discuss problems concerning the risk factors for CIM and CIP. RECENT FINDINGS: The pathophysiology of critical illness myopathy and critical illness polyneuropathy is complex, involving metabolic, inflammatory, and bioenergetic alterations. This review cites new evidence supporting several pathogenetic mechanisms. These include microvascular changes in peripheral nerves (with increased endothelial expression of E-selectin), the possible role for an altered lipid serum profile in promoting organ dysfunction (including nerve dysfunction), the damage or inhibition of complex I of the respiratory chain as a cause of muscle ATP depletion and bioenergetic failure, and the activation of specific intracellular proteolytic systems causing myofilament loss and apoptosis in CIM. The diagnostic role of direct muscle stimulation and the rapid quantification of myosin/actin ratio based on electrophoresis are also presented. SUMMARY: Basic and clinical research is unraveling the pathophysiological mechanisms of critical illness myopathy and polyneuropathy, and methods for rapid diagnosis are actively investigated. Future studies should better define the population at risk of developing CIM and CIP. In fact, although sepsis, multi-organ failure and steroids are often cited as risk factors, uncertainty remains due to the poor methodological quality of studies, or because of inferences that are exclusively based on animal studies. New simplified diagnostic techniques and machines for electrophysiological investigations of peripheral nerves and muscles in the intensive-care unit (ICU) patient would also be welcome. PMID- 15758593 TI - Multimodality monitoring and telemonitoring in neurocritical care: from microdialysis to robotic telepresence. AB - PURPOSE OF REVIEW: This review will highlight the state-of-the-art in brain monitoring in neurointensive care and define methods of integrating this technology into patient care using telemedicine methods. RECENT FINDINGS: Several new methods of brain monitoring have been established over the last several years including continuous EEG monitoring, brain tissue oxygenation, jugular venous oxygenation, and cerebral microdialysis. Observational research using these monitors has documented that the brain metabolism, blood flow and function are dynamic after a primary insult. The dynamic nature of the brain can predispose the brain to secondary insults that can occur in the setting of intensive care. Several variables of brain metabolism and function can be monitored and directly impact treatment decisions as well as provide diagnostic and prognostic information. General treatment guidelines for brain injury and brain hemorrhage were developed, in part, prior to implementation of use of these monitors, and there is a trend away from adoption of a one-size-fits-all approach and a trend towards monitor-guided therapy. Dealing with the data provided by multimodality monitoring can be overwhelming. Efficient use of such information requires methods to integrate diverse sets of information, and methods to access the online monitoring information remotely and at any time, day or night. Such remote access integration methods will be reviewed. SUMMARY: Multimodality and telemedicine techniques have advanced the state of knowledge about brain function in critically ill patients, and are presently being implemented to direct therapy. Increasing complexity of care will become commonplace, but will be facilitated by computer-enhanced tools that permit the intensivist to integrate this information into an improved treatment regimen. PMID- 15758594 TI - Non-neurological organ dysfunction in neurocritical care: impact on outcome and etiological considerations. AB - PURPOSE OF REVIEW: Organ dysfunction is an important determinant of outcome in critical care medicine. Patients with life threatening neurologic injury represent a distinct subset of critically ill patients in whom non-neurologic organ dysfunction may develop. In this paper the incidence and impact of non neurologic organ dysfunction in patients with major neurologic injury will be reviewed. Further, potential etiological considerations will be addressed and management strategies discussed. RECENT FINDINGS: Non-neurologic organ dysfunction is extremely common in patients with brain injury occurring in 80-90% of patients admitted to intensive-care units. Several studies have now identified this dysfunction as an independent predictor of poor outcome in neurocritical care. This dysfunction may arise as a result of the neurologic injury or secondary to treatment. Massive catecholamine release continues to be the primary etiological theory of non-neurologic organ dysfunction due to brain injury. Currently employed therapies directed at intracranial hypertension such as maintenance of cerebral perfusion pressure and the use of hypothermia or barbiturates predispose non-neurologic organ dysfunction. SUMMARY: Non-neurologic organ dysfunction is common. This dysfunction independently predicts poor outcome following brain injury and represents a potentially modifiable risk factor. Further study is required to develop optimal management strategies. PMID- 15758595 TI - Effects of selective decontamination of digestive tract on mortality and antibiotic resistance in the intensive-care unit. AB - PURPOSE OF REVIEW: Since its introduction in 1984 several small trials have studied the infection prevention regimen of selective decontamination of the digestive tract (SDD) in intensive care patients. Although meta-analyses of these studies suggested that SDD could reduce mortality, it continued to be a highly controversial strategy. There were not only serious doubts about the methodological quality of the meta-analyses, fear also existed that SDD would lead to increased antibiotic resistance. Recently, two new large randomized trials have been published that studied the effects of SDD on mortality and resistance. In this article, we will review the concept on which SDD is based and the present knowledge of the effects of SDD on mortality and antibiotic resistance. RECENT FINDINGS: In accordance with earlier meta-analyses of small studies, two recent randomized trials have confirmed that selective decontamination of the digestive tract significantly lowers mortality and decreases the emergence of antibiotic resistance. Limitation of these studies is the fact that they were conducted in intensive-care units (ICUs) with a low prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococcus (VRE). SUMMARY: There is convincing evidence that selective decontamination of the digestive tract (SDD) lowers mortality as well as resistance in circumstances with low prevalence of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant enterococcus (VRE). SDD should still be considered experimental in area's where MRSA and VRE are endemic. However, given the important potential benefits of SDD, more studies are urgently needed to adapt SDD in a way that proves effective in those settings. PMID- 15758596 TI - The 'small for size' liver syndrome. AB - PURPOSE OF REVIEW: Small-for-size syndrome (SFSS) is a clinical syndrome described following liver transplantation (LT) and extended hepatectomy. New evidence has emerged documenting the importance of preoperative evaluation of functional liver mass, liver quality, influence of portal hypertension, and variations in surgical technique to improve outcome. RECENT FINDINGS: SFSS is characterized by postoperative coagulopathy and liver dysfunction due to insufficient functional liver mass. Recent radiologic advances allow accurate preoperative estimation of total, graft, and remnant liver volume (RLV). In adult to-adult living donor liver transplantation (LDLT), a graft-to-recipient body weight ratio > or = 0.8% or graft weight ratio > or = 30% are important to avoid SFSS. Minimal functional RLV following extended hepatectomy is > or = 25% in a normal liver, and > or = 40% with preoperative liver dysfunction. Preoperative portal vein or hepatic artery embolization to increase RLV and function after extended hepatectomy, and the increasing use of parenchymal-sparing segmental resections have improved outcome. In LT, the evolving use of split livers, LDLT and marginal grafts has resulted in increased recognition of SFSS. This has led to a renewed interest in defining the pathophysiology, and the development of new surgical techniques to reduce its incidence. SUMMARY: Current radiologic imaging techniques can be used to evaluate liver volume and the risk of SFSS following LT and extended hepatectomy. Intraoperative techniques to predict postoperative dysfunction are emerging, and may be helpful in directing the use of pre-emptive surgical interventions. The future lies in the development of perioperative liver protection and support in predicted SFSS, and enhancement of healthy liver regeneration. PMID- 15758597 TI - Intra-abdominal hypertension in the critically ill: it is time to pay attention. AB - PURPOSE OF THE REVIEW: There has been an exponentially increasing interest in intra-abdominal hypertension (IAH). Comparison of the published data however is difficult due to the lack of consensus definitions. This review will focus on the available literature from the last 2 years. A Medline and PubMed search was performed using 'intra-abdominal pressure' (IAP), 'intra-abdominal hypertension' (IAH), and 'abdominal compartment syndrome' (ACS) as search items. The aim was to find an answer to the question 'Isn't it time to pay attention to intra-abdominal pressure in the critically ill?' RECENT FINDINGS: Although the number of studies published on this topic is steadily increasing and confirms the pathophysiologic implications of IAH on end-organ function within and outside the abdominal cavity it remains difficult to compare the literature data because the measurement methods and definitions used are not uniform. Provocative data have been published regarding the interactions between the abdominal and thoracic compartments especially in patients with capillary leak and fluid overload; most of this data raises even more questions than it gives answers and may therefore strengthen the nonbelievers who consider IAP, IAH and ACS as epiphenomena in critically ill patients. Unless the international scientific community does not come forward with clear-cut definitions we will keep comparing 'apples with oranges.' SUMMARY: It is time to pay attention to intra-abdominal pressure in the critically ill. It is also time for standardized IAP measurement methods, good consensus definitions and randomized interventional studies. PMID- 15758598 TI - Bibliography. Current world literature. Neuroscience. PMID- 15758600 TI - Thromboembolism in pregnancy. AB - PURPOSE OF REVIEW: Venous thromboembolism is the leading cause of maternal death in the UK. Thrombophilia underlies many thrombotic disorders in pregnancy. The high prevalence of thrombophilic defects in the population, the association of defects with venous thromboembolism and the special considerations for management make it a widely debated subject. RECENT FINDINGS: A limited number of studies measuring the risk of venous thromboembolism in pregnancy with thrombophilia have been conducted within the last year. Studies confirm that heritable thrombophilias are associated with increased risk of venous thromboembolism in pregnancy. However, estimated risks vary between individual studies. The risk of venous thromboembolism with acquired thrombophilia remains unclear. Guidelines have been published to guide clinicians in preventing and treating venous thromboembolism in pregnancy; however, large-scale, randomized controlled trials need to be conducted to establish the effectiveness of administering antithrombotic agents in pregnancy. Although selective thrombophilia screening based on prior history of venous thromboembolism has been proposed, the overall clinical and economic benefit of universal and selective screening is unsupported. SUMMARY: Due to the lack of studies, gaps still exist in our knowledge of the risk of pregnancy-related venous thromboembolism associated with thrombophilia. In particular, accurate estimates are required for the risks of acquired thrombophilias. Furthermore, the true effectiveness of anticolagulants in pregnancy needs to be established through well-conducted studies and randomized controlled trials. These studies will inform clinicians and help to determine the optimum management and prevention strategies for thrombophilia and venous thromboembolism in pregnancy. PMID- 15758601 TI - HIV in pregnancy. AB - PURPOSE OF REVIEW: This review discusses recent articles on various aspects of the prevention of mother-to-child transmission during pregnancy and delivery. RECENT FINDINGS: Rapid human immunodeficiency virus (HIV) testing of women in labour whose status is not known allows the prompt treatment of mother and baby to reduce transmission risk. The feared clinical resistance in the mother after treatment with a single dose of nevirapine has been confirmed. Strategies are required to minimize this resistance and allow the use of nevirapine for treatment of the mother. There are new findings of mitochondrial toxicity in babies who have been exposed to anti-retroviral medicines during pregnancy or delivery, but the clinical implications are not clear. Long-term follow-up of exposed children is required. Resource-poor countries are starting to use multiple drugs to further reduce transmission to the infant. These efforts are reducing the rates of transmission to the level found in affluent countries. SUMMARY: Improvements in treatment continue to reduce the risk of HIV transmission from mother to child in resource-poor countries, but subsequent maternal resistance continues to be a problem since treatment for the mother's health is now possible. The long-term effects on the infant are still not understood. PMID- 15758602 TI - Thyroid disorders in pregnancy. AB - PURPOSE OF REVIEW: There is now increasing understanding of the association between not only overt, but also subclinical, thyroid disorders and dysfunction with adverse reproductive outcome. In particular, hypothyroidism and thyroid autoimmunity disorders have been shown to have both short- and long-term consequences on mother and child. An updated review is merited to revise many of the traditional views on thyroid disorders in pregnancy. RECENT FINDINGS: Thyroid disorders constitute the commonest group of pre-gestational endocrine disorders found in pregnant women. In mothers taking antithyroid medications, breastfeeding is considered safe. The relatively high prevalence of hypothyroidism, especially subclinical hypothyroidism, the significance of screening and treatment, and the roles of iodine insufficiency and thyroid antibodies on the outcome of pregnancy and long-term neurological development of the offspring have been documented. In hypothyroid women, the dose of thyroxine replacement often needs to be adjusted from as early as the first trimester to maintain an adequate circulating thyroxine concentration. SUMMARY: Apart from overt hyperthyroidism and hypothyroidism diagnosed before and during pregnancy, biochemical abnormalities or evidence of thyroid autoimmunity in clinically euthyroid women can affect both obstetric outcome and long-term neurological development of the offspring. Screening for thyroid function and autoimmunity, and timely and appropriate treatment, will improve pregnancy outcome. The thyroid function of infants born to mothers with thyroid disorders should also be assessed as serial monitoring and treatment may be necessary. PMID- 15758603 TI - Thalassaemia screening in pregnancy. AB - PURPOSE OF REVIEW: This review provide an update on antenatal screening and diagnosis of thalassaemia disorders. RECENT FINDINGS: The topics covered are the effectiveness of antenatal screening programmes for thalassaemia, its prenatal diagnosis, molecular basis and laboratory findings, ultrasound screening for haemoglobin Bart's disease, and non-invasive prenatal diagnosis of thalassaemia. SUMMARY: Universal antenatal screening for thalassaemia carriers should be implemented in populations with a high prevalence of this condition. The appropriate measure to screen for alpha and beta thalassaemias remains mean cell haemoglobin (<27 pg) or mean corpuscular volume (<80 fl). A haemoglobin pattern and iron profile should follow if the red cell indices are low. In a population where alpha thalassaemia is prevalent, it is advisable to check the partner's mean cell haemoglobin or mean corpuscular volume as well. Further cascades of investigations will depend on these results and the prevalence of other haemoglobinopathies in that population. Invasive prenatal diagnosis remains the gold standard for diagnosis in high-risk couples. Provided expertise is available, ultrasound measurement of the cardiothoracic ratio appears a good screening tool for alpha thalassaemia major. Non-invasive prenatal diagnosis by identification of a paternal mutation in maternal plasma, although currently at the experimental stage, may be an option in the future. PMID- 15758604 TI - Intrauterine growth restriction. AB - PURPOSE OF REVIEW: Intrauterine growth restriction remains one of the major problems in obstetrics. Recent published literature on this problem is summarized in this review. RECENT FINDINGS: Intrauterine growth restriction contributes disproportionately to neonatal mortality and morbidity in both preterm and term babies, and is a predisposing factor to major psychiatric sequelae such as depression, suicide and suicidal attempts. More evidence is accumulating to show that fetal Doppler changes of the ductus venosus and umbilical vein are good surrogate markers for fetal academia. The timing of delivery remains controversial, however. The Growth Restriction Intervention Trial showed that delayed delivery in those up to 30 weeks may be associated with lower rates of cerebral palsy and Griffiths development quotient under 70. In dichorionic twins, selective fetocide of one severe intrauterine growth restriction fetus in midtrimester twin pregnancies complicated by severe preeclampsia may abort the disease process and prolong the pregnancy. For monochorionic twins, the finding of intermittent absent or reversed end diastolic flow in the umbilical artery may be a manifestation of the transmission of the bi-directional waveforms of arterio arterial anastomosis, but has been shown to be associated with an increased risk of intrauterine death in the growth restricted fetus and brain damage in the larger fetus. SUMMARY: The timing of delivery of the preterm growth restricted fetus remains controversial. Intrauterine growth restriction with intermittent absent or reversed end diastolic flow in the umbilical artery of monochorionic twins poses difficulties in assessment. PMID- 15758605 TI - Management of pregnancy and labour in cases diagnosed with major fetal malformation. AB - PURPOSE OF REVIEW: To explore the obstetrical management of pregnancies with severe fetal malformations, taking into account the rapid improvement in ultrasound technology and interventional possibilities. RECENT FINDINGS: Unique studies using modern ultrasound techniques describe the evolution of spina bifida and the subsequent varying development of hydrocephalus. In addition, the dilated fetal stomach is shown to be an ominous sign of complications in cases of gastroschisis. Such complications can be decreased by prenatal supervision using cardiotocography, which detects signs of fetal distress. Recent studies confirm previous findings that caesarean section does not improve the outcome in cases of fetal gastroschisis or spina bifida. SUMMARY: Recent research has demonstrated the development of severe fetal malformations. New studies have shown the positive effects of fetal supervision in cases of gastroschisis. Furthermore, caesarean section is not beneficial compared with vaginal delivery in such cases. Fetal surgery is still undergoing rapid development and has the potential to improve outcome in the near future. PMID- 15758606 TI - Fetal electrocardiogram waveform analysis in labour. AB - PURPOSE OF REVIEW: ST-waveform analysis of the fetal electrocardiogram (ECG) has emerged from experimental and observational studies to clinical use based on the outcome of two large randomized controlled trials and a European Union-supported project on the structured dissemination of knowledge and experience by establishing regional centres of excellence. The review focuses on the outcome from the host of studies and those recently published. RECENT FINDINGS: The database is dominated by a Swedish randomized controlled trial demonstrating not only improved outcome with regard to cord-artery metabolic acidosis and fewer operative interventions for non-reassuring fetal state but, most importantly, the marked and significant reduction in the risk of neonates showing signs of moderate or severe neonatal encephalopathy. The first report from the European Union project is presented, verifying the clinical relevance of the STAN methodology. The outcome of the large European Union project of 8000 deliveries confirms the observations of the randomized controlled trials. A study on the outcome shows improvements in accuracy and consistency among clinicians when automated ST analysis is added to the fetal heart rate trace. SUMMARY: After years of dedicated research, it appears as though ST analysis of the fetal ECG has become an additional source of information allowing detailed analysis of fetal responses and more accurate identification of a non-reassuring fetal status. The technology provides continuous information throughout labour. As with any new methodology, structured efforts on training and user feedback are required to fully implement the STAN methodology in clinical practice. PMID- 15758607 TI - Acute tocolysis. AB - PURPOSE OF REVIEW: Emergency uterine relaxation may decrease the morbidity and mortality of the mother and her fetus. Obstetricians need to be aware of the indications, pharmacological methods, efficacy and complications of acute tocolysis. RECENT FINDINGS: A variety of pharmacological agents are used to suppress uterine contractions. Newer agents like cyclo-oxygenase-2 inhibitors (Celecoxib) and oxytocin antagonists (atosiban) have been introduced into clinical practice with the hope of reducing the complications of betasympathomimetic drugs. Calcium-channel blockers are used but there are recent case reports of acute pulmonary oedema with the use of these agents. Most of the trials on tocolytics have been for suppression of preterm labour. Nitroglycerin has been used successfully as an acute tocolytic during Caesarean sections and manual removal of placenta. A recent randomized trial has suggested that atosiban may be an option for acute intrapartum tocolysis. This article will review the recent literature on the use of pharmacological agents used to suppress uterine contractions in emergency obstetric situations. SUMMARY: Acute tocolysis may be indicated in antepartum, intrapartum and postpartum periods for a variety of indications. It may help reduce maternal and fetal morbidity and mortality. The ideal tocolytic is yet to be developed. Research is needed to develop a drug which has a greater uterospecificity with no effect on other organs with a rapid onset and a short duration of action. PMID- 15758608 TI - Maternal collapse. AB - PURPOSE OF REVIEW: This review will summarize the latest publications on the causes, prevention and treatment of maternal collapse during pregnancy and the postpartum period. It will also explore some future trends particularly in the area of education and team working. RECENT FINDINGS: Active management of the third stage of labour has been shown to decrease the incidence of postpartum haemorrhage. Inadequate or prolonged resuscitation following major postpartum haemorrhage can lead to myocardial damage. Guidelines that are followed and the use of drills and simulators may increase effective team working and lead to earlier recognition and treatment of maternal collapse. Some cases of amniotic fluid embolism resemble anaphylaxis and should be treated as such. Important evidence-based guidelines on the management of antithrombolytic prophylaxis and the management of pulmonary embolism have recently been published, which should help clinicians clarify their medical care plans. New resuscitation guidelines in the UK will probably mean that all healthcare workers (including obstetricians and midwives) will need appropriate training every year. SUMMARY: Understanding the causes of maternal collapse, its early recognition and prompt resuscitation should decrease both maternal and fetal morbidity and mortality. There is increasing evidence that effective preventative measures and improved multidisciplinary team working may have an important impact on maternal and fetal well-being. PMID- 15758609 TI - Too much information or not enough? Editorial review. PMID- 15758610 TI - Genetic screening and diagnosis. AB - PURPOSE OF REVIEW: To review the latest developments in screening and diagnosis of non-chromosomal genetic diseases. RECENT FINDINGS: Major recent advances include the completion of the Human Genome Project, the use of microarray and related technologies for mass screening and diagnosis of thousands of genetic abnormalities, and non-invasive prenatal diagnosis using fetal DNA in maternal plasma. SUMMARY: The rapid development in molecular biological technologies makes it possible to screen and to diagnosis thousands of genetic conditions, mutations and also predispositions to chronic diseases or traits, either prenatally or after birth. Clinical application of non-invasive prenatal diagnosis using fetal DNA in maternal plasma has become a reality. The arrival of the molecular genetic era also leads to many new ethical, social and medico-legal problems and dilemmas that obstetricians will have to face in the near future. There is an urgent need for the development of a new model for provision of genetic screening and diagnosis. PMID- 15758611 TI - Detection of chromosomal abnormalities by comparative genomic hybridization. AB - PURPOSE OF REVIEW: Comparative genomic hybridization (CGH) is a modified in-situ hybridization technique. In this type of analysis, two differentially labeled genomic DNAs (study and reference) are cohybridized to normal metaphase spreads or to microarray. Chromosomal locations of copy number changes in the DNA segments of the study genome are revealed by a variable fluorescence intensity ratio along each target chromosome. Thus, CGH allows detection and mapping of DNA sequence copy differences between two genomes in a single experiment. RECENT FINDINGS: Since its development, comparative genomic hybridization has been applied mostly as a research tool in the field of cancer cytogenetics to identify genetic changes in many previously unknown regions. It is also a powerful tool for detection and identification of unbalanced chromosomal abnormalities in prenatal, postnatal and preimplantation diagnostics. SUMMARY: The development of comparative genomic hybridization and increase in resolution analysis by using the microarray-based technique offer new information on chromosomal pathologies and thus better management of patients. PMID- 15758612 TI - Preimplantation diagnosis: a realistic option for assisted reproduction and genetic practice. AB - PURPOSE OF REVIEW: Preimplantation genetic diagnosis (PGD) allows genetically disadvantaged couples to reproduce, while avoiding the birth of children with targeted genetic disorders. By ensuring unaffected pregnancies, PGD circumvents the possible need and therefore risks of pregnancy termination. This review will describe the current progress of PGD for Mendelian and chromosomal disorders and its impact on reproductive medicine. RECENT FINDINGS: Indications for PGD have expanded beyond those used in prenatal diagnosis, which has also resulted in improved access to HLA-compatible stem-cell transplantation for siblings through preimplantation HLA typing. More than 1000 apparently healthy, unaffected children have been born after PGD, suggesting its accuracy, reliability and safety. PGD is currently the only hope for carriers of balanced translocations. It also appears to be of special value for avoiding age-related aneuploidies in in-vitro fertilization patients who have a particularly poor prognosis for a successful pregnancy; the accumulated experience of thousands of PGD cycles strongly suggests that PGD can improve clinical outcome for such patients. SUMMARY: PGD would particularly benefit poor prognosis in-vitro fertilization patients and other at-risk couples by improving reproductive outcomes and avoiding the birth of affected offspring. PMID- 15758613 TI - Ultrasound and fetal diagnosis. AB - PURPOSE OF REVIEW: The purpose of this review is to highlight publications from the last year that have advanced the use of ultrasound in obstetrics. RECENT FINDINGS: Anatomic examination of the fetus in the first trimester has been emphasized because it allows for early diagnosis of many conditions. The prevalence of absent nasal bone, a marker for trisomy 21, in euploid fetuses depends on ethnicity. Nasal bone hypoplasia is another marker for Down syndrome. Studies on genetic screening in the first trimester have involved various serum analytes, adjustments in timing and calculations, use in multiple gestations, and the association of extreme measurements with adverse outcomes. A first-trimester integrated screening approach, which incorporates nuchal translucency, nasal bone, crown-rump length, pregnancy-associated plasma protein-A, and free beta human chorionic gonadotropin, has the potential to maximize detection rates of Down syndrome and trisomy 18 and minimizes the screen-positive rate. The value of combining first and second-trimester results in sequential, contingent, or integrated screening protocols has been assessed. Isolated mild ventriculomegaly (10-12 mm) may prove to be a normal variant, and the role of 'soft' ultrasound markers in genetic counseling continues to be debated. Anomaly or high-risk status detection in the second trimester has been enhanced by the use of Doppler, 3D/4D ultrasound, and magnetic resonance imaging. SUMMARY: Imaging techniques have been critical in the development of screening methods for Down syndrome or trisomy 18 and for euploid fetuses at high risk for adverse outcomes. PMID- 15758614 TI - Chorionic villus sampling and amniocentesis. AB - PURPOSE OF REVIEW: The advantages and disadvantages of common invasive methods for prenatal diagnosis are presented in light of new investigations. RECENT FINDINGS: Several aspects of first-trimester chorionic villus sampling and mid trimester amniocentesis remain controversial, especially fetal loss rate, feto maternal complications, and the extension of both sampling methods to less traditional gestational ages (early amniocentesis, late chorionic villus sampling), all of which complicate genetic counseling. A recent randomized trial involving early amniocentesis and late chorionic villus sampling has confirmed previous studies, leading to the unquestionable conclusion that transabdominal chorionic villus sampling is safer. The old dispute over whether limb reduction defects are caused by chorionic villus sampling gains new vigor, with a paper suggesting that this technique has distinctive teratogenic effects. The large experience involving maternal and fetal complications following mid-trimester amniocentesis allows a better estimate of risk for comparison with chorionic villus sampling. SUMMARY: Transabdominal chorionic villus sampling, which appears to be the gold standard sampling method for genetic investigations between 10 and 15 completed weeks, permits rapid diagnosis in high-risk cases detected by first trimester screening of aneuploidies. Sampling efficiency and karyotyping reliability are as high as in mid-trimester amniocentesis with fewer complications, provided the operator has the required training, skill and experience. PMID- 15758616 TI - EUS in the management of the patient with dysplasia in Barrett's esophagus. AB - Barrett's esophagus is the most important risk factor in the development of adenocarcinoma of the esophagus. Barrett's esophagus is generally regarded as the most significant complication of gastroesophageal reflux disease. Adenocarcinoma occurs more frequently in the setting of high-grade dysplasia. The prognosis of adenocarcinoma of the esophagus is strongly correlated with the stage of disease. The prognosis of late stage disease is extremely poor. Cure may be achieved when disease is found at an early stage. Esophagectomy has been the definitive treatment of limited stage adenocarcinoma of the esophagus. The morbidity and mortality rate for esophagectomy is high. Therefore, alternative endoscopic methods for curative treatments have gained popularity. The two main endoscopic therapies, photodynamic therapy and endoscopic mucosal resection, are both effective when applied to early-stage disease. Traditional evaluation of the patient with Barrett's esophagus with high-grade dysplasia includes esophago gastro-duodenoscopy (EGD) with biopsy and computed tomography of the chest. Endoscopic ultrasound (EUS) has gained popularity in the evaluation of the patient with Barrett's esophagus and high-grade dysplasia because it is the only imaging technique capable of delineating the separate histologic layers of the gastrointestinal tract. The principal role of EUS in evaluating patients with Barrett's-associated dysplasia is to identify patients who may be candidates for endoscopic ablative (endoscopic mucosal resection, photodynamic therapy) therapies. EUS has been shown to be superior to computed tomography (including high resolution spiral CT) or magnetic resonance imaging for preoperative staging in patients with high-grade dysplasia and carcinoma. This review of the literature summarizes the ability of EUS to evaluate patients with Barrett's esophagus and high-grade dysplasia. PMID- 15758617 TI - Cost minimization in endoscopy center scheduling: a case-controlled study. AB - BACKGROUND: Traditional endoscopy center scheduling often results in nonurgent inpatient endoscopic procedures being delayed until late in the day and can prolong length of hospital stay and costs. We report the first controlled study designed to evaluate the effect of an early morning fast-track triage endoscopy unit on the cost and length of stay of a general GI inpatient population. METHODS: A case-control methodology matched a cohort of patients undergoing morning triage procedures with historical controls that underwent standard add-on scheduling endoscopy. Outcome indices and patient quality of care measures were compared between cases and controls. RESULTS: Analysis of patients most likely to benefit from rapid endoscopy showed significant advantage comparing fast-track endoscopy patients to controls in time to endoscopy (0.63 vs. 1.00 days, P = 0.01), length of stay (1.22 vs. 1.78 days, P = 0.05), and hospital costs (2,793 dollars vs. 3,586 dollars, P = 0.02). CONCLUSIONS: When routine endoscopy is the rate-limiting step for hospital discharge in the general GI patient, early morning scheduling with a reserved time and space for inpatient endoscopy is a cost-minimizing factor in a busy endoscopy center that may save significant hospital costs while preserving optimal patient outcomes. PMID- 15758618 TI - The yield of bleeding scans in acute lower gastrointestinal hemorrhage. AB - GOALS/BACKGROUND: Previous studies have demonstrated low yield and conflicting results as to the ability of technetium-99m labeled red blood cell (TRBC) scintigraphy to accurately localize the site of bleeding in acute lower gastrointestinal hemorrhage. Our objective was to evaluate the accuracy of TRBC scintigraphy in clinical day-to-day practice at our institution and to determine predictors of a positive test. METHODS: A retrospective medical records review of all patients who underwent TRBC scintigraphy over a 5-year period was conducted at a single academic medical center. The site of bleeding found on TRBC scintigraphy was correlated with the endoscopic, angiographic, surgical, and pathologic findings in each patient. RESULTS: A total of 127 scans were performed in 115 patients. Forty-nine scans were positive (39%) and 78 were negative (61%). Forty-two patients had further evaluation after a positive scan, and the bleeding site was confirmed to be accurate in 20 of 42 patients (48%). A contradictory bleeding site was found in 5 of 49 positive scans (10%). Patients transfused greater than 2 units of packed red blood cells within 24 hours preceding the TRBC scan were twice as likely to have a positive scan (64%) than those transfused 2 units or less (32%) (P = 0.002). Multivariate analysis demonstrated that the number of units of blood transfused within 24 hours prior to the TRBC scan and the lowest recorded hematocrit were significantly different between patients with positive and negative scans. There was no difference in the rate of endoscopy between patients with a positive or negative scan result. CONCLUSIONS: TRBC scintigraphy has a relatively low yield in the evaluation of acute lower gastrointestinal bleeding. Objective selection criteria such as number of units of packed red blood cells transfused prior to obtaining the scan may increase the overall yield. PMID- 15758619 TI - Use of reagent strips for the rapid diagnosis of spontaneous bacterial empyema. AB - GOALS: To assess the utility of reagent strips for rapid diagnosis of spontaneous bacterial empyema in cirrhotic patients with hepatic hydrothorax. BACKGROUND: Analysis of ascitic fluid using reagent strips is a useful diagnostic test for spontaneous bacterial peritonitis. METHODS: A reagent strip for leukocyte esterase designed for the testing of urine was used to evaluate pleural fluid analysis in 47 nonselected thoracenteses in 28 cirrhotic patients with hepatic hydrothorax. RESULTS: Twelve spontaneous bacterial empyemas were diagnosed. Simultaneous spontaneous bacterial peritonitis was present in 7 of 10 cases in which ascites fluid was analyzed. When a test result of 3 or 4 was considered positive, sensitivity was 83% (10 of 12), specificity was 100% (35 of 35), and positive predictive value was 100%. When result of 2 or more was considered positive, sensitivity was 92% (11 of 12), specificity was 80% (28 of 35), and negative predictive value was 97%. CONCLUSION: Analysis of pleural fluid with reagent strips is a rapid, easy to use, and inexpensive tool for the diagnosis of spontaneous bacterial empyema in cirrhotic patients. A positive result should be considered an indication for antibiotic therapy. PMID- 15758620 TI - A polymorphism of the bactericidal/permeability increasing protein (BPI) gene is associated with Crohn's disease. AB - BACKGROUND: The bactericidal/permeability increasing protein (BPI) is involved in the elimination of gram-negative bacteria. A functionally relevant single nucleotide polymorphism of the BPI gene causes an amino acid exchange (Glu216Lys). STUDY: To evaluate whether this single nucleotide polymorphism contributes to the predisposition to inflammatory bowel disease, we compared the allele frequencies of 265 patients with Crohn's disease, 207 patients with ulcerative colitis, and 608 healthy controls. RESULTS: The Glu/Glu genotype frequency was decreased significantly in Crohn's disease patients as compared with controls (P < 0.027). No differences were obvious in patients with ulcerative colitis. CONCLUSIONS: Failure of the innate intestinal immune system could be involved in the pathogenesis of Crohn's disease via reduced/impaired defense against gram-negative bacteria. PMID- 15758621 TI - Efficacy and safety of endoscopic balloon dilation of symptomatic upper and lower gastrointestinal Crohn's disease strictures. AB - GOALS: To evaluate efficacy and safety of endoscopic balloon dilation with or without intralesional steroid injection, of symptomatic upper gastrointestinal (UGI) and lower gastrointestinal (LGI) Crohn's disease (CD) strictures. BACKGROUND: Patients with CD commonly develop obstructive symptoms secondary to gastrointestinal strictures. When these do not resolve with medical management, surgery is usually the only alternative. Limited data are available on the safety and efficacy of endoscopic through-the-scope (TTS) balloon dilation of CD strictures. STUDY: We performed a retrospective review of TTS balloon dilations done on CD-related UGI and LGI strictures. Postdilation intralesional steroid injections were done at the discretion of the endoscopist. Stricture was defined as luminal stenosis <10 mm in diameter, through which a scope could not be passed. Technical success was defined as the ability of the scope to traverse the stricture postdilation. Long-term success rate was claimed if a patient remained asymptomatic and did not require surgery or further endoscopic dilation. RESULTS: Over 4 years, we performed 29 stricture dilations on 17 patients (10 female, 7 male) with 20 strictures. The mean follow-up period was 18.8 months (range, 5-50 months). Stricture locations were as follows: rectal, 5; sigmoid colon, 2; colo colonic anastomosis, 3; ileocolonic anastomosis, 4; ileum, 1; descending colon, 1; cecum, 1; and distal duodenal bulb, 3 patients. Technical success was achieved in 28 of 29 stricture dilations (96.5%). Ten strictures (34.5%) were dilated to <15 mm and 19 (65.5%) to > or = 15 mm diameter. Long-term success rate in the <15 mm group was 70%, and in > or = 15 mm group was 68.4%. Four quadrant steroid injections were done on 11 strictures. The recurrence rate in this group was 10% and that in the nonsteroid group was 31.3%. The overall long-term success rate was 76.5% by intent-to-treat analysis. Three perforations occurred (all colonic) during 29 stricture dilations, a complication rate of 10% with no mortalities. CONCLUSION: We report the first series of TTS balloon dilations with or without intralesional steroid injection, of both primary and anastomotic UGI and LGI strictures in CD patients. Long-term success was achieved in 76.5% patients with a complication rate of 10%. This mode of therapy appears safe and effective and can be considered as an alternative to surgery in selected patients with medically refractory CD-associated GI strictures. Success rates were better in patients who received four quadrant steroid injections. No difference was seen in stricture recurrence rate or complications based on diameter of TTS balloon used. PMID- 15758622 TI - Topical treatment of distal active ulcerative colitis with beclomethasone dipropionate or mesalamine: a single-blind randomized controlled trial. AB - GOALS: Therapy for active ulcerative colitis (UC) usually involves rectal formulations of corticosteroids (CS), which are characterized by the risk of systemic steroid-related adverse effects. BACKGROUND: To compare the efficacy and safety of the topically acting CS beclomethasone dipropionate (BDP) versus mesalamine (5-ASA) in the treatment of active UC. STUDY: Patients with mild to moderate distal active UC were randomized to a 6-week treatment with BDP 3 mg enema o.d. or 5-ASA 1 g enema daily in a single-blind, multicenter, parallel group, controlled study. The primary efficacy variable was the decrease in Disease Activity Index (DAI) score. Safety variables were adrenal function, monitoring of adverse events, vital signs, and laboratory parameters. RESULTS: A total of 217 patients were enrolled and treated with BDP (n = 111) or 5-ASA (n = 106). A significant decrease in the DAI score (P < 0.05) was observed in both treatment groups, with a clinical remission rate of 36.7% in the BDP group and of 29.2% in the 5-ASA group. Both treatments were well tolerated. No changes from baseline in morning cortisol levels were observed in the BDP group. CONCLUSIONS: BDP administered as a rectal enema over a 6-week treatment period was efficacious and safe in patients with active UC, without interference with pituitary adrenal axis. PMID- 15758623 TI - Deficiencies in knowledge and familial risk communication among colorectal adenoma patients. AB - GOALS: Our primary objectives were to assess knowledge about familial risk and risk communication among colorectal adenoma patients. BACKGROUND: The first degree relatives (FDRs) of colorectal adenoma patients diagnosed before the age of 60 years may be at increased risk of colorectal cancer and should begin screening by the age of 40 years. Adherence to this recommendation requires effective risk communication between physicians, affected patients, and their at risk relatives. STUDY: We conducted a telephone survey of 129 consecutive English speaking adenoma patients younger than 60 years treated by 11 endoscopists at two medical centers. Information regarding patient demographics, awareness of familial risk, and risk communication was elicited. RESULTS: A total of 71 eligible subjects responded to the survey. Few responders (n = 25, 33%) were aware that their FDRs were at increased risk of colorectal cancer, and only 56% of knowledgeable patients identified a physician as the source of information. Awareness was strongly associated (P < or = 0.001) with patient-initiated information-seeking behavior. Most knowledgeable patients (n = 20, 80%) reported that they had informed > or = 1 FDRs about their diagnosis, and most (68%) felt that it was the patient's responsibility to notify at-risk relatives. CONCLUSIONS: Most colorectal adenoma patients younger than 60 years are unaware of the familial implications of their diagnosis and therefore unlikely to notify at-risk FDRs. PMID- 15758624 TI - Association of anti-tumor necrosis factor therapy with the development of multiple sclerosis. AB - A 35-year-old woman with a history of indeterminate colitis developed symptoms of multiple sclerosis after treatment with infliximab. Neurologic examination confirmed upper and lower extremity motor and sensory deficits. MRI showed multiple enhancing white matter lesions distributed throughout her brain as well as her thoracic spine. There may be a link between inflammatory demyelinating disease of the central nervous system and anti-tumor necrosis-alpha therapy. This case report describes the onset or worsening of a demyelinating process after the initiation of infliximab therapy in a patient with indeterminate colitis. PMID- 15758625 TI - Cystic fibrosis: an overview. AB - Cystic fibrosis (CF) is one of the most common inherited disorders of white populations. The isolation and cloning of the gene in CF that encodes the production of a transport protein that acts as an apical membrane chloride channel, termed cystic fibrosis transmembrane conductance regulator (CFTR), have improved our understanding of the disorder's pathophysiology and has aided diagnosis, but has also revealed the disease's complexity. Gene replacement therapy is still far from being used in patients with CF, mostly because of difficulties in targeting the appropriate cells. Life expectancy of patients with this disorder has greatly improved over past decades because of better symptomatic treatment strategies. This article summarizes advances in understanding and treatment of CF. PMID- 15758626 TI - Hepatic copper in patients receiving long-term total parenteral nutrition. AB - GOALS: To assess the possibility of iatrogenic hepatic copper overload in adult patients on long-term total parenteral nutrition (TPN). BACKGROUND: TPN predisposes to hepatic copper accumulation through disturbances of the enterohepatic bile acid pool, but iatrogenic copper overload through TPN solutions may occur as well. STUDY: Quantitative hepatic copper and multiple clinical, biochemical, and histopathologic parameters were compared between patients with long-term TPN associated liver disease (n = 28) and patients with drug-induced cholestatic liver disease (n = 10). RESULTS: Eighty-nine percent of TPN patients and all controls had mildly elevated hepatic tissue copper, but 29% of TPN patients had levels above the diagnostic threshold for Wilson's disease. Quantitative hepatic copper correlated positively with serum aspartate aminotransferase (P = 0.001, r = 0.59), total bilirubin (P < 0.001, r = 0.65), and direct bilirubin (P < 0.001, r = 0.63) in TPN patients, but not in controls. The amount of hepatic copper did not correlate with the duration of TPN (median, 1.9 years; range, 0.3-18.0 years) or serum copper levels. TPN patients with significant cholestasis accumulated more copper than patients with no or only minimal cholestasis (P = 0.002). CONCLUSIONS: Significant hepatic copper overload in TPN patients occurs through chronic cholestasis in TPN-associated liver disease and is independent from the total duration of TPN. Iatrogenic copper overload through trace elements in TPN solutions does not seem to be a significant factor. PMID- 15758628 TI - Prospective randomized trial of post-liver biopsy recovery positions: does positioning really matter? AB - GOALS: The goal of this study was to compare three commonly practiced post-liver biopsy recovery postures with respect to pain levels and overall patient acceptability. BACKGROUND: Percutaneous liver biopsy is a valuable procedure used in the diagnosis, prognosis, and treatment of liver diseases. No standardized recovery posture exists, and no studies have been performed to compare these different recovery techniques. STUDY: Ninety adult patients were randomized into three arms of the study: right-side (R), supine (S), and combination (C) groups. A validated Visual Analogue Scale (VAS) was given to patients to grade the pain and discomfort experienced during recovery intervals as well as to grade the overall acceptability of the biopsy experience. RESULTS: Immediately following liver biopsy, more pain was experienced by patients randomized to R with a mean VAS score of 26.5 of 100, compared with 14.2 (P = 0.026) and 12.1 (P = 0.009) for C and S groups, respectively. At the end of recovery, there was no difference among the three groups. The mean acceptability score was 89.2 of 100 for C versus 94.5 for S (P = 0.047) and 94.8 for R (P = 0.046). CONCLUSION: This study is the first to examine differing post-liver biopsy techniques. When three commonly practiced recovery positions, C, R, and S, are compared, C was the least acceptable position. Patients should be placed in position R or S during recovery. PMID- 15758627 TI - Acute upper gastrointestinal bleeding in patients with liver cirrhosis and in noncirrhotic patients: epidemiology and predictive factors of mortality in a prospective multicenter population-based study. AB - GOALS: To assess epidemiologic features and predictive factors of mortality of acute upper gastrointestinal bleeding (UGIB). STUDY: During a 6-month period, a prospective population-based study including all the UGIB occurring in a geographic area of 3 million people was conducted. Data from cirrhotic patients were compared with those of noncirrhotic patients. RESULTS: A total of 2,133 UGIB were recorded, 21.9% in cirrhotic patients (n = 468). Endoscopic hemostasis was performed in 46.5% and 8.3% in cirrhotic and noncirrhotic patients, respectively (P < 0.001). Mortality during hospitalization was 23.5% in cirrhotic patients and 11.2% in noncirrhotic patients (P < 0.001). Six independent predictive factors of mortality were observed in both patient groups: a prothrombin level less than 40%, an UGIB occurring in inpatients, a concomitant digestive carcinoma, a hematemesis revealing the UGIB, a recent use of steroid drugs, and age over 60 years. Four other predictive factors of mortality were also identified in noncirrhotic patients. CONCLUSIONS: Although epidemiologic features, clinical course, management, and prognosis of UGIB were quite different in cirrhotic and noncirrhotic patients, the majority of predictive factors of mortality were the same in both patient groups. These data underline the major role of debilitated status and hepatic failure in the prognosis of UGIB in cirrhotic patients. PMID- 15758629 TI - Octreotide therapy for advanced hepatocellular carcinoma. AB - Treatment options for advanced hepatocellular carcinoma (HCC) remain limited. Recently, octreotide has been proposed for therapy, although its efficacy remains controversial. Thus, the aim of this open-label pilot study was to evaluate the response of HCC to long-acting octreotide (Sandostatin LAR). Thirty patients were enrolled for this prospective 2-year trial. Initially, patients were given short acting octreotide to ensure drug tolerability. Thereafter, patients received long acting octreotide 30 mg IM every 4 to 6 weeks. Measurable disease was assessed at 3-month intervals. Five of 30 patients were unable to tolerate the test dose, and 1 patient was reevaluated and underwent hepatic resection. The remaining 24 patients, who received long-acting octreotide, all had advanced stage of disease with multifocal-massive morphology (67%), vascular thrombosis (63%), or extrahepatic spread (17%), but well compensated liver disease. The treatment was well tolerated, except for diarrhea. Median time to tumor progression was 3.6 months, and median survival was 5.1 months. Seven patients (29%) had stable disease (median duration of 8.0 months) with 2 patients demonstrating disease stability for 24 months. In conclusion, although occasional patients appear to have stable disease on long-acting octreotide therapy, overall the beneficial response in terms of time to tumor progression and survival is limited. PMID- 15758631 TI - Interferon for metastatic renal cell cancer causing regression of hereditary hemorrhagic telangiectasia. PMID- 15758630 TI - Fulminant hepatic failure secondary to neoplastic infiltration of the liver. AB - Two patients with previously normal liver function, who presented with fulminant hepatic failure (FHF) of unknown etiology despite an extensive evaluation, are described. No etiology for FHF was apparent with initial evaluation. One patient was found to have nearly complete replacement of hepatic parenchyma by metastasis from an occult small cell lung carcinoma identified postmortem. The other patient had lymphomatous infiltration of the liver detected by a liver biopsy. Imaging studies were performed in the patients and did not reveal any evidence of neoplastic infiltration of the liver. Neoplastic involvement of liver should be considered in the differential diagnosis of FHF of unknown etiology. The imaging studies in this setting can be misleading. PMID- 15758632 TI - Braised liver with herbs: the risks of naturopathic hepatoprotection. PMID- 15758645 TI - Hepatocellular transporters and cholestasis. AB - The secretion of bile is the result of active hepatocellular transport processes, most of which occur across the canalicular membrane of liver cells. Disturbance of the function and/or expression of these transporters leads to the intracellular accumulation of toxic bile acids, thereby promoting cholestatic liver cell injury. Genetically determined alterations of hepatobiliary transporter function are increasingly recognized as important risk factors for an individual's susceptibility to develop cholestasis. It has become evident that, besides the established pathogenic role of mutations in canalicular transporter genes in progressive and benign forms of familial intrahepatic cholestasis, genetics may also play an important role in acquired cholestatic syndromes, such as intrahepatic cholestasis of pregnancy or drug-induced cholestasis. This overview summarizes the physiologic function and regulation of human hepatobiliary transport systems and discusses the impact of their genetic variations for the pathophysiology of different cholestatic syndromes. PMID- 15758646 TI - Molecular regulation of hepatobiliary transport systems: clinical implications for understanding and treating cholestasis. AB - Hepatobiliary transport systems are responsible for hepatic uptake and excretion of bile salts and other biliary constituents (eg, bilirubin) into bile. Hereditary transport defects can result in progressive familial and benign recurrent intrahepatic cholestasis. Exposure to acquired cholestatic injury (eg, drugs, hormones, proinflammatory cytokines, biliary obstruction or destruction) also results in altered expression and function of hepatic uptake and excretory systems, changes that may maintain and contribute to cholestasis and jaundice. Recruitment of alternative efflux pumps and induction of phase I and II detoxifying enzymes may limit hepatic accumulation of potentially toxic biliary constituents in cholestasis by providing alternative metabolic and escape routes. These molecular changes are mediated by bile salts, proinflammatory cytokines, drugs, and hormones at a transcriptional and posttranscriptional level. Alterations of hepatobiliary transporters and enzymes are not only relevant for a better understanding of the pathophysiology of cholestatic liver diseases, but may also represent important targets for pharmacotherapy. Drugs (eg, ursodeoxycholic acid, rifampicin) used to treat cholestatic liver diseases and pruritus may counteract cholestasis via stimulation of defective transporter expression and function. In addition, therapeutic strategies may be aimed at supporting and stimulating alternative detoxification pathways and elimination routes for bile salts in cholestasis. PMID- 15758647 TI - Portal hypertension: from bedside to bench. AB - The initial factor leading to portal hypertension is an increase in hepatic resistance. Later, an increase in portal blood flow contributes to maintain and exacerbate portal hypertension despite the development of portosystemic collaterals. The critical step in the development and acceptance of these concepts, which proved crucial for the management of patients with portal hypertension, was the development of animal models. These allowed the full characterization of the profound hemodynamic abnormalities in the systemic and splanchnic circulation associated with portal hypertension, and the elucidation of the molecular mechanisms implicated in these disturbances. This review traces how seminal clinical observations in the 1950s raised meaningful questions that were subsequently answered at the bench, leading to our current understanding of the pathophysiology of portal hypertension and of the pathogenesis of severe complications of cirrhosis, such as variceal bleeding or ascites. PMID- 15758648 TI - Increased hepatic resistance: a new target in the pharmacologic therapy of portal hypertension. AB - Increased resistance to portal blood flow is the primary factor in the pathophysiology of portal hypertension, and is mainly determined by the morphologic changes occurring in chronic liver diseases. This is aggravated by an increased hepatic vascular tone, which results from an insufficient hepatic bioavailability of nitric oxide (NO) and an increased production of circulating and local vasoconstrictors (angiotensin, endothelin, cysteinyl-leukotrienes, and thromboxane, among others). This dynamic and reversible component provides the rationale for the use of therapies aimed at decreasing portal pressure by reducing the vascular tone. Among them, systemic and liver-selective NO donors, statins, and gene therapy with adenovirus encoding NO synthases have been used to increase NO availability with promising results. Other attempts have been the blockade of the effect of vasoconstrictors, using anti alpha-adrenergic agents and renin-angiotensin system blockers. Some of these pharmacologic approaches have already been incorporated into clinical practice while others are still under investigation. PMID- 15758649 TI - Mechanisms of pulmonary vascular complications of liver disease: hepatopulmonary syndrome. AB - Pulmonary vascular abnormalities occurring in the setting of liver disease have been increasingly recognized as important clinical entities that influence survival and liver transplant candidacy in affected patients. The most common such abnormality, the hepatopulmonary syndrome, is found in 15% to 20% of patients with cirrhosis. These disorders have no effective medical therapies. Experimental models of hepatopulmonary syndrome have identified a sequence of hepatic and pulmonary endothelial alterations that lead to nitric oxide and carbon monoxide-mediated intrapulmonary vasodilatation. A key role for shear stress-mediated pulmonary endothelial endothelin B receptor overexpression and cholangiocyte ET-1 production and release has emerged as a mechanism for local nitric oxide production in the lung. How these alterations are influenced by bacterial translocation and the systemic hyperdynamic circulatory state and whether similar changes occur in human disease are areas of ongoing investigation. PMID- 15758650 TI - Inhibition of albumin synthesis in chronic diseases: molecular mechanisms. AB - The albumin gene is expressed specifically in the liver after birth, and this expression is regulated predominantly at the transcriptional level. Regulatory proteins occupy specific DNA sequences within the promoter and enhancer of the albumin gene. The interaction between the CCAAT/enhancer binding protein (C/EBP) beta and the albumin DNA is critical for albumin synthesis. Cachexia-induced hypoalbuminemia is mediated by tumor necrosis factor (TNF)-alpha. In turn, TNF alpha stimulates oxidative stress, NO synthesis, and phosphorylation of C/EBP beta within its nuclear localization signal (NLS). Consequently, C/EBP-beta is exported from the nucleus, preventing it to act as a transcriptional factor on the albumin gene. Antioxidants, NOS inhibitors. and dominant negative, nonphosphorylatable C/EBP-beta peptides block phosphorylation of C/EBP-beta within the NLS and its nuclear export as well as rescue the abnormal albumin gene expression, suggesting potential therapeutic interventions. PMID- 15758651 TI - Mild hypothermia for acute liver failure: a review of mechanisms of action. AB - Brain edema with intracranial hypertension is a major complication in patients with acute liver failure. Current therapies for this complication include a variety of pharmacologic and interventional measures, some of which are frequently associated with adverse effects or contraindications. Even though these measures usually allow the control of intracranial hypertension for a certain period of time, recurrence is common. New therapies are therefore needed. Increasing clinical and experimental evidence suggests that induction of mild hypothermia (32 degrees C-35 degrees C) may be a therapeutic alternative. Similar to traumatic brain injury or brain stroke, induction of mild hypothermia seems highly effective to reduce intracranial pressure in patients with acute liver failure. Several mechanisms by which mild hypothermia may prevent brain edema and intracranial hypertension in this condition have been disclosed and may include beneficial effects on ammonia metabolism, as well as on the disturbances of brain osmolarity, cerebrovascular hemodynamics, brain glucose metabolism, inflammation, and others. Improvement of systemic hemodynamics and amelioration of liver injury may be other benefits of the systemic induction of mild hypothermia, but the impact of potential adverse events, such as infection, should also be taken into account. At a time when mild hypothermia is increasingly used in several specialized centers, performance of a randomized controlled trial seems critical to confirm the benefits of mild hypothermia in acute liver failure and to provide adequate guidelines for its use. PMID- 15758652 TI - The role of matrix stiffness in hepatic stellate cell activation and liver fibrosis. AB - Liver fibrosis results primarily from the action of hepatic stellate cells, nonparenchymal cells of the liver that undergo transdifferentiation into fibrogenic, proliferative, and contractile myofibroblasts. Stellate cell transdifferentiation has been modeled by the culture of primary cells, a system that has yielded important information about factors determining the phenotype of these cells. Recent evidence suggests that the growth factor TGF-beta (acting through the cytoplasmic signaling intermediate Smad3) and the mechanical properties of the underlying matrix play particularly important roles in hepatic stellate cell transdifferentiation and that this transdifferentiation is a multistep process. The interrelationship between TGF-beta and matrix stiffness and the implications of the in vitro findings for liver fibrosis are now the subject of intensive investigation and will likely lead to important insights into the diagnosis and treatment of liver disease. PMID- 15758653 TI - Mitochondrial dysfunction in hepatitis C. AB - Chronic hepatitis C induces a state of hepatic oxidative stress that is more pronounced than that present in many other inflammatory liver diseases. This review summarizes recent information that the hepatitis C virus (HCV) core protein plays an important role in this phenomenon. Core protein localizes to mitochondria, particularly at the points of contact between mitochondrial outer membrane and endoplasmic reticulum. Its expression causes inhibition of electron transport at complex I, increased complex I reactive oxygen species (ROS) production, decreased mitochondrial glutathione, and increased mitochondrial permeability transition in response to exogenous oxidants and tumor necrosis factor-alpha. Possible mechanisms of the core protein effects include direct interaction with electron carriers and indirect effects mediated by changes in mitochondrial calcium. These results suggest that antioxidant approaches may prove beneficial for patients with chronic hepatitis C. PMID- 15758655 TI - Quality incentive payment systems: promise and problems. AB - Public and corporate pressure to improve the quality of healthcare in the United States has never been greater. Rising costs, recent discouraging appraisals of patient safety, accelerating malpractice litigation, and an increasing burden of chronic disease have intensified the demand for a change in current policies. Central to these efforts are payers who are increasingly creating financial incentives for providers to deliver high-quality healthcare. It is hoped that these programs (sometimes referred to as "quality incentive payment systems") will stimulate the adoption of systems that will reduce variability and assure that high-quality care is being delivered. Whether these programs will achieve their objectives remains to be determined. There are many unanswered questions about their effectiveness. PMID- 15758654 TI - Molecular studies of liver disease in erythropoietic protoporphyria. AB - GOALS: The goal of this study was to define molecular determinants of liver disease in erythropoietic protoporphyria (EPP). BACKGROUND: EPP is a genetic disorder in which deficient ferrochelatase activity causes excessive production of protoporphyrin, which is excreted in bile. Some patients develop liver disease that necessitates transplantation. STUDY: Ferrochelatase gene analysis was done in 25 families with EPP to identify mutations and a polymorphism (IVS3-48c) that causes low gene expression. Expression of multiple hepatic genes was also examined by DNA microarray analysis in patients who had liver transplantation to identify genes with altered regulation. RESULTS: Heterozygous ferrochelatase mutations were found in 43 individuals. In 94% of 31 symptomatic patients, 15 of whom had liver disease, the polymorphism was also present in the nonmutant allele. Explanted liver of patients who had transplantation showed significant change in expression of several genes involved in wound healing, organic anion transport, and oxidative stress. CONCLUSIONS: Patients with EPP who develop liver disease usually have a mutation in one ferrochelatase allele that alters enzyme function, together with a polymorphism in the nonmutant allele that causes low gene expression. This results in significant increase in the hepatobiliary excretion of protoporphyrin, which can damage the liver through both cholestatic injury and oxidative stress. PMID- 15758656 TI - 50 years of leadership of the Yale Digestive Disease Program. PMID- 15758657 TI - Surveillance and screening for Barrett esophagus and adenocarcinoma. AB - Current recommendations for screening and surveillance of Barrett esophagus and related lesions are based on recent guidelines by the Practice Parameters Committee of the American College of Gastroenterology. The purpose of this review is to critically examine the rationale and evidence behind these recommendations. There is strong rationale for vigorous initial testing to document the baseline status and identify early adenocarcinoma, and for surveillance of high-grade dysplasia. Recommendations for esophagectomy in patients with high-grade dysplasia need to be individualized. However, recommendations for surveillance of low-grade dysplasia and specialized intestinal metaplasia without dysplasia are largely opinion statements not well supported by objective data. Although cancers identified by surveillance are at earlier stages than those diagnosed without prior endoscopic evaluation, surveillance failures are common. Recommendations for screening and surveillance are not evidence-based and unlikely to alter national mortality from esophageal adenocarcinoma. Their impact on individual patients depends on individual circumstances. Current recommendations are limited by inconsistent endoscopic findings and sampling errors, inconsistent histologic diagnoses of Barrett esophagus and dysplasia, and our poor understanding of the natural history of various histologic lesions. Future directions include validation of methods that reduce these inconsistencies by in vivo detection of abnormalities and by objective diagnostic markers besides grades of dysplasia, such DNA content analysis and molecular markers, and improved understanding of the disease progression. Effective screening programs depend on development of simple, inexpensive, and reliable methods to identify the small group of patients truly at high risk for adenocarcinoma for endoscopic screening. PMID- 15758658 TI - Motor and sensory function of the esophagus: revelations through ultrasound imaging. AB - Catheter based high frequency intraluminal ultrasound (HFIUS) imaging is a powerful tool to study esophageal sensory and motor function and dysfunction in vivo in humans. It has provided a number of important insights into the longitudinal muscle function of the esophagus. Based on the ultrasound images and intraluminal pressure recordings, it is clear that there is synchrony in the timing as well as the amplitude of contraction between the circular and the longitudinal muscle layers of the esophagus in normal subjects. On the other hand, in patients with spastic disorders of the esophagus, there is an asynchrony of contraction related to the timing and amplitude of contraction of the two muscle layers during peristalsis. Achalasia, diffuse esophageal spasm, and nutcracker esophagus (spastic motor disorders of the esophagus) are associated with hypertrophy of the circular as well as longitudinal muscle layers. A sustained contraction of the longitudinal muscle of the esophagus is temporally related to chest pain and heartburn and may very well be the cause of symptoms. Longitudinal muscle function of the esophagus can be studied in vivo in humans using dynamic ultrasound imaging. Longitudinal muscle dysfunction appears to be important in the motor and sensory disorders of the esophagus. PMID- 15758659 TI - Pathogenesis of diarrhea in ulcerative colitis: new views on an old problem. AB - BACKGROUND: Whereas water movement into the intestinal lumen occurs secondary to Cl secretion in secretory diarrheal diseases, defects in key transport processes lead to profound decreases in colonic Na, Cl, and water absorption in ulcerative colitis. STUDIES AND RESULTS: Recent studies indicate reduced expression/activity of apical Na channels and basolateral Na, K-ATPase, leading to loss of electrogenic Na absorption in the distal colon and rectum. There is also likely to be a decrease in electroneutral NaCl cotransport, which is present throughout the colon. Preliminary work on basolateral K channel abundance and activity in colonic epithelial cells suggests that whole-cell K conductance is decreased in ulcerative colitis, leading to epithelial cell depolarization, and further limitation of Na absorption. In addition, there is a marked reduction in colonic epithelial resistance, which reflects a decrease in the integrity of intercellular tight junctions and the presence of apoptotic foci. CONCLUSIONS: Impaired Na and Cl transport, combined with enhanced epithelial "leakiness," results in a profound decrease in the capacity of the inflamed colon to absorb salt and water. Transport abnormalities in ulcerative colitis may at least partly reflect the effects of proinflammatory cytokines, raising the possibility of novel approaches to the restoration of colonic absorptive capacity in this disease. PMID- 15758660 TI - Bicarbonate secretion: a neglected aspect of colonic ion transport. AB - Understanding of the mechanism of colonic electrolyte transport has markedly increased over the past three decades. This article provides a brief summary of the critical features of Na, Cl, and K transport in the large intestine and how these processes may be altered in diarrhea. Less understood is the mechanism of colonic HCO3 secretion. Recent progress in the regulation of HCO3 secretion in the distal colon is summarized with emphasis on the interrelationship between Cl dependent, short-chain fatty acid (SCFA)-dependent, and cAMP-induced HCO3 secretion. cAMP down-regulates Cl-dependent HCO3 secretion, while SCFA stimulates HCO3 secretion but also inhibits both Cl-dependent and cAMP-induced HCO3 secretion. As SCFAs are the primary anions in stool, it is likely that SCFA dependent HCO3 secretion is the primary mechanism of HCO3 secretion in the mammalian colon. Future studies will undoubtedly provide increased understanding of the mechanism of HCO3 secretion in health and disease. PMID- 15758661 TI - Past and current theories of etiology of IBD: toothpaste, worms, and refrigerators. AB - While tremendous advances have improved the understanding of inflammatory bowel disease, with regard to environmental risk factors as well as the biochemical nature of the inflammatory process, a determination of primary etiology remains elusive. Numerous theories have been proposed in the past century concerning the cause of Crohn's disease and ulcerative colitis with implications for specific therapies. On further study, most of these ideas and therapies have failed to be accurate in theory or therapeutic approach. Others remain untested or are the focus of current investigation and controversy. This paper reviews the dominant theories of primary etiology. These hypotheses include infectious causes such as Mycobacteria paratuberculosis and measles. Allergic and nutritionally related causes have been the focus of considerable research. Microparticles, which is part of the concept behind toothpaste as a cause, have been suggested more broadly to be the principal factor initiating Crohn's disease. Several of these concepts rely on the idea that there is an increased intestinal permeability that is the central defect leading to Crohn's disease. Rather than being an excessive T cell driven process, Crohn's has been suggested to be an innate immune deficiency, leading to the use of colony stimulating factors to augment the intestinal barrier function and innate immunity. A variety of changes in the gut flora, ranging from a basic dysbiosis to the absence of helminths, have been proposed as the root cause of inflammatory bowel disease. PMID- 15758662 TI - Preventing neoplastic progression in ulcerative colitis. AB - Patients with ulcerative colitis (UC) and Crohn's colitis carry an increased risk for developing colorectal cancer (CRC). Patients with more extensive colitis, greater duration of disease, concomitant primary sclerosing cholangitis, and a family history of CRC are at greatest risk among UC patients. Young age at disease onset and greater inflammatory burden have also been proposed as risk factors. Maneuvers that limit the impact of cancer in colitis include prophylactic colectomy, which is unacceptable to most UC and Crohn's colitis patients, and dysplasia surveillance. Although recommended in a number of practice guidelines, surveillance has not yet been demonstrated to reduce CRC mortality or morbidity. A number of factors, including low levels of agreement among pathologists interpreting surveillance specimens, patients lost to follow up, failure to recommend colectomy once dysplasia has been discovered, and others, hinder the success of surveillance. In an effort to compensate for the limitations of surveillance, chemoprevention and newer endoscopic and molecular techniques are being assessed for their effectiveness in augmenting or replacing conventional surveillance. PMID- 15758663 TI - Reduced CFTR function and the pathobiology of idiopathic pancreatitis. AB - Idiopathic chronic pancreatitis (ICP) is the leading cause of chronic pancreatitis in children and nonalcoholic adults. The risk of developing ICP is increased in individuals who have mutations of the cystic fibrosis gene (CFTR) and of a trypsin inhibitor gene (PSTI). In studies from the United States and France, the risk of ICP is increased about 40-fold by having two abnormal copies of the CFTR gene, about 14-fold by having the N34S PSTI mutation, and about 500 fold by having both. When ICP patients have two abnormal copies of the CFTR gene, there is also evidence of reduced residual CFTR protein function in extrapancreatic tissues based on clinical findings and nasal ion transport responses. Thus, pancreatitis risk is highest in individuals who have abnormalities in both the pancreatic ducts (CFTR) and acini (PSTI). These findings indicate that PSTI is a modifier gene for CFTR-related ICP and have implications for the diagnosis and pathogenesis of pancreatitis. PMID- 15758664 TI - Epithelial differentiation in pancreatic development and neoplasia: new niches for nestin and Notch. AB - Changes in epithelial differentiation represent a characteristic early feature of human pancreatic cancer. Recent work suggests that many of these changes may reflect a pathologic recapitulation of developmental events. Reflecting this principle, metaplastic and neoplastic pancreatic epithelium appear to share many features in common with embryonic pancreatic epithelium, including reactivation of the Notch signaling pathway. In this review, we summarize recent studies involving regulation of epithelial differentiation in both embryonic and adult pancreas and highlight the role of Notch in regulating an exocrine progenitor pool. PMID- 15758665 TI - Drug-induced hepatotoxicity: 2005. AB - The removal from the marketplace of several widely prescribed drugs due to hepatotoxicity has attracted considerable attention. Now under extensive review are means by which we can better identify hepatic risk prior to federal approval. Assessment of risk-to-benefit ratios regarding a novel agent with hepatotoxicity issues (especially one for a life-threatening condition) requires considerable judgment and education on the part of prescribers and patients. The spectrum of drug-induced liver injury is broad with simulation of almost all unknown liver disorders. Drug-induced liver injuries often have a somewhat characteristic signature, as regards type of injury (hepatocellular vs cholestatic) and time of onset. The diagnosis of drug-induced liver injury is often one of exclusion with initial suspicion based on circumstantial evidence. Factors affecting susceptibility to drug-induced injury include age, sex, concomitant use of other drugs, and genetic polymorphism in metabolic pathways involved in activation or disposition of therapeutic drugs. Drug-drug interactions present particular problems in patients, often elderly, who are receiving several drugs simultaneously. Mechanisms of drug-induced liver injury are many and varied. With many drugs, intermediary products produced during metabolism are highly reactive and toxic. In these situations, the balance between the rate of production of the metabolite and the effectiveness of the drug may determine whether or not hepatic injury occurs. PMID- 15758666 TI - Pathophysiology of cholangiopathies. AB - The diseases of the intrahepatic biliary tree are a large group of potentially evolutive congenital and acquired liver disorders affecting both the adult and pediatric populations. They represent a relevant cause of liver-related morbidity and mortality and an important indication for liver transplantation, particularly in children. While the practical approach to patients affected by biliary tree diseases has not significantly changed yet, the conceptual approach to the pathophysiology of cholangiopathies has witnessed important advances that will be discussed. The primary cell target of the pathogenetic sequence of these disorders is the biliary epithelium. Cholangiocytes have multifaceted functions, not limited to bile production. Their capability to secrete a range of different pro-inflammatory mediators, cytokines, and chemokines indicates a major role of cholangiocytes in the inflammatory reaction. Furthermore, paracrine secretion of growth factors and peptides mediates an extensive cross-talk with other liver cell types, including hepatocytes, stellate, and endothelial and inflammatory cells. Cholangiopathies share a number of pathogenetic mechanisms, including inflammation, cholestasis, fibrosis, apoptosis, altered development, and neoplastic transformation. These basic disease mechanisms will be discussed in detail, along with the distinct features of a number of cholangiopathies. Furthermore, an increase in the biliary cell compartment is a common response to many forms of liver injury, from cholangiopathies to viral and fulminant hepatitis. Elucidation of these pathophysiologic mechanisms will likely provide clues for future therapeutic strategies. Furthermore, understanding the role of cholangiocytes in liver regeneration/repair and the mechanisms of cholangiocyte activation and their relationship with liver progenitor cell will be of further interest. PMID- 15758667 TI - Impact of smoking on fracture healing and risk of complications in limb threatening open tibia fractures. AB - OBJECTIVES: Current data show smoking is associated with a number of complications of the fracture healing process. A concern, however, is the potential confounding effect of covariates associated with smoking. The present study is the first to prospectively examine time to union, as well as major complications of the fracture healing process, while adjusting for potential confounders. SETTING: Eight Level I trauma centers. PATIENTS: Patients with unilateral open tibia fractures were divided into 3 baseline smoking categories: never smoked (n = 81), previous smoker (n = 82), and current smoker (n = 105). OUTCOME MEASURE: Time to fracture healing, diagnosis of infection, and osteomyelitis. METHODS: Survival and logistic analyses were used to study differences in time to fracture healing and the likelihood of developing complications, respectively. Multivariate models were used to adjust for injury severity, treatment variations, and patient characteristics. RESULTS: After adjusting for covariates, current and previous smokers were 37% (P = 0.01) and 32% (P = 0.04) less likely to achieve union than nonsmokers, respectively. Current smokers were more than twice as likely to develop an infection (P = 0.05) and 3.7 times as likely to develop osteomyelitis (P = 0.01). Previous smokers were 2.8 times as likely to develop osteomyelitis (P = 0.07), but were at no greater risk for other types of infection. CONCLUSION: Smoking places the patient at risk for increased time to union and complications. Previous smoking history also appears to increase the risk of osteomyelitis and increased time to union. The results highlight the need for orthopaedic surgeons to encourage their patients to enter a smoking cessation programs. PMID- 15758668 TI - Low incidence of ulnar nerve injury with crossed pin placement for pediatric supracondylar humerus fractures using a mini-open technique. AB - OBJECTIVES: Several recent studies have suggested that medial pinning in pediatric supracondylar humerus fractures leads to increased rates of ulnar nerve injury. The purpose of this study was to determine the risk of iatrogenic ulnar nerve injury in a consecutive series of supracondylar fractures treated using a standardized technique of crossed pin placement. DESIGN: Single cohort retrospective. SETTING: Metropolitan university tertiary care center. PATIENTS AND PARTICIPANTS: Seventy-one consecutive children with Gartland type II or type III supracondylar humerus were treated surgically by 2 pediatric orthopaedic surgeons at 1 institution between 1995 and 2000 using a medial mini-open and cross-pinning technique. Sixty-five patients were available for follow-up (92%). INTERVENTION: Patients were treated with a combination of medial and lateral pins using a mini-incision technique. MAIN OUTCOME MEASUREMENTS: Outcomes analyzed included ulnar nerve injury and clinical and radiographic evidence of healing. RESULTS: The study group consisted of 65 patients, of whom 29 (45%) presented with Gartland type III fractures, and the remaining 36 (55%) presented with a type II fracture. There were no ulnar nerve motor injuries. One patient was noted to have transient sensory changes in the ulnar nerve distribution postoperatively, which resolved by the 1-week follow-up visit. All patients were noted to have normal ulnar motor and sensory nerve function at final follow-up (average 4.5 months). No cases of nonunion, malunion, or infection were identified during the follow-up period. CONCLUSIONS: The rate of iatrogenic ulnar nerve injury with this specific technique of crossed pin placement for extension type supracondylar humerus fractures was extremely low in this series. A single case of transient ulnar sensory neuropraxia occurred. Our series demonstrates that crossed pin fixation can be performed safely and reliably and is an appropriate treatment option for unstable supracondylar humerus fractures. PMID- 15758669 TI - Midterm results of treatment with a retrograde nail for supracondylar periprosthetic fractures of the femur following total knee arthroplasty. AB - OBJECTIVES: Although the short-term results of supracondylar periprosthetic fractures treated with retrograde nailing have been satisfactory, there is always a concern about the long-term survival of the prosthesis. The aim of the study was to evaluate fracture healing and knee functional outcome with a follow-up time of at least 2 years in periprosthetic fractures of the knee treated with a supracondylar nail. DESIGN: Cohort study. PATIENTS: There were 9 patients with 10 periprosthetic fractures. In 1 patient, the fracture occurred intraoperatively. In the others, the time between the total knee arthroplasty and the periprosthetic fracture ranged between 2 weeks and 7 years (average time: 2.78 years). The mean follow-up was 34.5 months (25-52 months). MAIN OUTCOME MEASUREMENTS: The Western Ontario and McMaster Universities index was used to evaluate the functional result postoperatively using the paired t test as the statistical test. Fracture union was assessed with plain x-rays. RESULTS: All the fractures united within 3 months. One fracture united in extreme valgus (35 degrees) and was revised to a stemmed total knee replacement. There were no infections and no prosthesis loosening. The paired t test before the fracture and after the operation demonstrated no statistically significant differences; however, there was a trend toward lower functional score postoperatively. CONCLUSIONS: It appears that retrograde nailing is a reliable technique to treat periprosthetic supracondylar fractures. It provides adequate stability until fracture union. The morbidity of the operation is minimal, and the complication rate is low. The midterm results in our study showed that none of the prostheses required revision. In our opinion, it is the treatment of choice for a periprosthetic fracture when the prosthesis is stable. PMID- 15758670 TI - Knee arthrodesis with simultaneous lengthening using the Ilizarov method. AB - OBJECTIVE: To determine whether knee arthrodesis with simultaneous lengthening using the Ilizarov method for a nonreconstructable knee joint with bone loss and infection is a successful salvage procedure. DESIGN: Retrospective review of patients. SETTING: University hospital-based orthopaedic practice. PATIENTS: From 1999 to 2001, 4 consecutive patients with a nonreconstructable knee joint, bone loss, and infection after trauma underwent knee arthrodesis with simultaneous lengthening. INTERVENTION: Arthrodesis of the knee with simultaneous limb lengthening through an osteotomy of the tibia and/or femur and the use of an Ilizarov frame. External bone stimulation was used at the knee arthrodesis site and the lengthening sites. Application of this device began during the early distraction phase and continued until frame removal. MAIN OUTCOME MEASURES: Bony union at the arthrodesis and bone lengthening sites, alignment of the lower extremity, limb length discrepancy, infection, pain, and outcome scales (SF-36 scores and American Academy of Orthopaedic Surgeons lower limb modules). RESULTS: Bony union of the knee arthrodesis and lengthening sites and good alignment were achieved in all 4 patients. Mean amount of lengthening was 5.4 cm (range 2.5-11.5 cm). Average time in frame was 11 months (range 6-17 months). Limb length discrepancy after treatment averaged 1.8 cm (range 0.6-3.7 cm). Mean duration of follow-up after frame removal was 35 months (range 28-48 months). At follow-up, infection had not recurred, pain was not present, and assistive devices were not needed for ambulation. Average SF-36 scores improved in all 8 categories, and the average American Academy of Orthopaedic Surgeons lower limb modules improved from a mean of 33 (range 11-37) to a mean of 68 (range 51-76). CONCLUSION: Knee arthrodesis with simultaneous lengthening can be performed successfully using the Ilizarov method. It enables surgeons to optimize limb length during knee arthrodesis. The use of external fixation and the avoidance of internal implants may be advantageous in the presence of or history of infection. The Ilizarov frame provides stability that allows weight bearing during treatment. PMID- 15758671 TI - The effect of transfixion wire number and spacing between two levels of fixation on the stiffness of proximal tibial external fixation. AB - OBJECTIVES: Anatomic constraints about the proximal tibia limit the ability to insert wires at the biomechanically optimum angle of 90 degrees, thus diminishing the potential stability of external fixators used for proximal tibial fractures. To overcome this problem, surgeons use more than 2 wires at a single level of fixation or a second level of fixation. This study evaluated the effect of transfixion wire number and placement of a second level of fixation on the stiffness of proximal tibial external fixation. METHODS: A fiberglass tibia fixed into an idealized ring external frame was tested. Load-deformation behavior was compared among the different wire numbers (1 level with 2, 3, 4, and 5 wires) and placement of a second level of fixation (2 wires first level and 1 wire second level, and 2 wires first level and 2 wires second level) at 2, 3, 4, 5, 6, 7, 8, 9, and 10 cm distance from the first level of fixation. Identical loading conditions of central axial compression, medial compression-bending, posterior compression-bending, posteromedial compression-bending, and torsion were used. Stiffness values were calculated from the load-displacement and the torque-angle curves. RESULTS: An increase in wire number at 1 level led to an overall increase in stiffness, whereas the addition of a second level of fixation and increased spread between these 2 levels increased bending stiffness. CONCLUSIONS: The addition of a second level of external fixation of the proximal tibia fragment with maximum possible distance between the 2 levels increases bending stiffness, whereas increasing the number of transfixion wires increases overall stiffness. Axial and torsional stiffness is proportional to the total number of wires regardless of the number of levels of fixation. PMID- 15758672 TI - A biomechanical comparison of a dorsal 3.5-mm T-plate and a volar fixed-angle plate in a model of dorsally unstable distal radius fractures. AB - OBJECTIVES: To compare the biomechanical stability of internal fixation of extra articular, dorsally unstable distal radius fractures fixed by 1 of 2 methods, either a standard dorsal nonlocked T-plate or a volar locked fixed-angle plate. DESIGN: Biomechanical cadaveric study. SETTING: Biomechanical testing laboratory. INTERVENTION: In 6 matched pairs of fresh-frozen cadaveric specimens, a simulated unstable extra-articular distal radius fracture was created. The fractures were stabilized with either a dorsal 3.5-mm stainless steel T-plate or a titanium locked volar fixed-angle plate. Specimens were axially loaded at 5 points (centrally, volarly, dorsally, radially, and ulnarly) and then cyclically loaded for 5000 cycles with an 80 N central load. Postcyclical loading, specimens were once again axially loaded at the 5 points. MAIN OUTCOME MEASURES: Initial fixation stiffness and stiffness after midaxial cyclical loading was compared at the 5 points. RESULTS: With the volar locked fixed-angle plate, fixation was significantly stiffer than with the dorsal nonlocked T-plate for ulnar and volar loading in single-cycle testing. After cyclic loading, the locked volar fixed angle plate maintained more of its initial stiffness than the dorsal nonlocked T plate. The dorsal 3.5-mm stainless steel T-plate's stiffness when dorsally loaded significantly decreased after cyclical loading. CONCLUSIONS: The volar locked fixed-angle plate maintained a greater percentage of its initial stiffness after cyclic loading compared to the dorsal nonlocked plate. Also, the volar locked plate was stiffer than the dorsal nonlocked plate for all loading configurations tested except when subjected to a dorsally applied eccentric load. PMID- 15758673 TI - Operative treatment of displaced lateral malleolar fractures: the Knowles pin technique. AB - OBJECTIVES: To assess the use of Knowles pin fixation for isolated displaced lateral malleolar fractures and to present our surgical experience using this method. DESIGN: Retrospective evaluation. SETTING: The study was carried out at a municipal teaching hospital. The senior surgeons were all orthopedic trauma subspecialists. PATIENTS: A total of 168 patients meeting our inclusion criteria, an isolated displaced lateral malleolar fracture surgically treated by a Knowles pin between 1995 and 2000, were studied. All the patients had a stable syndesmosis, less than 50% comminution, and had no other operations in the same extremity. Patients were assigned into 4 groups according to the Orthopedic Trauma Association classification of ankle fractures. INTERVENTION: Open reduction and internal fixation with a Knowles pin fixation of the lateral malleolus. MAIN OUTCOME MEASUREMENTS: Functional results were evaluated using the Baird and Jackson ankle scoring system. RESULTS: There was a 100% union rate. The average satisfactory outcome of the 4 groups was 88.1%. No instrumentation problems were encountered. Three complications occurred, but resolved with nonoperative therapy. CONCLUSIONS: Knowles pin fixation for displaced lateral malleolar fractures is a useful and effective method. This implant offers several advantages, including easy application, less soft tissue dissection, less palpable instrumentation, stable fixation, and a short operating time. PMID- 15758674 TI - Repair or reattachment of the meniscus after fixation of a tibial plateau fracture. AB - The intact meniscus provides protection for healing of tibial plateau fractures, prevention of early posttraumatic arthritis, and maintenance of knee stability. However, tibial plateau fractures may have associated meniscal injury that may impair this function. In addition, the meniscotibial ligaments are often divided during submeniscal arthrotomy for exposure. Repair or reattachment of the meniscus can be difficult. We describe the technique of using Kirschner wire holes in proximal tibial plates for anchoring the meniscus. PMID- 15758675 TI - Temporary external fixation across the ankle after tibial nailing. AB - We describe the application of a temporary spanning external fixation device across the ankle in conjunction with intramedullary tibial nailing. This technique can be useful in selected patients with open fractures associated with severe soft-tissue trauma, skin grafts, or muscle flaps. The external fixator allows for wound access and keeps the foot and ankle in a neutral position preventing equinus. A brief period of rigid ankle immobilization is beneficial in preventing muscle motion and sheer stresses on flaps and skin grafts. The external fixator is removed at 3 to 6 weeks once the soft tissues have healed. PMID- 15758676 TI - Knee dislocation with ipsilateral femoral shaft fracture: a report of five cases. AB - We report the management and outcome of 4 patients with 5 knee dislocations associated with ipsilateral femoral shaft fractures. All patients were managed by immediate reduction of the knee dislocation, intramedullary nailing of the femur, and angiography, followed by postoperative immobilization of the knee (brace or external fixation) for a minimum of 6 weeks. Four of the 5 dislocations underwent a secondary ligament reconstruction. At the 2-year follow up, the mean Knee Society Score was 133 (range 99-170). PMID- 15758677 TI - Surgical management of a long segmental defect of the humerus using a cylindrical titanium mesh cage and plates: a case report. AB - This is a case report of a patient who sustained multiple gunshots resulting in a Gustilo Anderson type IIIB mid-shaft humeral fracture associated with extensive segmental bone and soft-tissue loss. The patient was treated initially by multiple irrigations, wound debridement, and a unilateral external fixator. After the soft tissue healed without infection, the mid-shaft humeral defect of approximately 8 cm segmental bone loss was reconstructed with a cylindrical titanium mesh cage packed with a composite of cancellous allograft and demineralized bone matrix putty and stabilized with limited contact dynamic compression plates. At 13 months follow-up, plain radiographs demonstrated a healed construct with good alignment, and computed tomography images demonstrated bony in-growth through the cage. The patient had full range of motion at the shoulder and the elbow. This technique may be a reasonable alternative when treating large segmental bone defects of the humerus. PMID- 15758679 TI - Osteochondral fragment removal from the hip joint: Opinion: observation. PMID- 15758678 TI - Autologous osteochondral mosaicplasty for treatment of a posttraumatic defect of the lateral tibial plateau: a case report with two-year follow-up. AB - Posttraumatic osteochondral defects following a tibial plateau fracture are common and a serious complication that may lead to the development of posttraumatic arthrosis. Successful reconstruction of the tibial plateau must include restoration of limb alignment, repair of bone defects, restoration of the articular cartilage, and preservation of the menisci. When osteochondral defects are present, the use of bulk bone grafts to restore the original articular surface anatomy of the tibial plateau is difficult due to incongruity between the graft and the original joint surface. Recognizing this, an autologous osteochondral transplantation utilizing the mosaic technique was performed successfully on a 32-year-old male alpine skier with a posttraumatic osteochondral defect following a tibial plateau fracture. At 2 years postsurgery, the patient had regained the capacity to perform most activities of daily living and to participate in sports. Clinical examination revealed an improvement of the Lysholm score from 48 points to 72 points. PMID- 15758699 TI - Early stage prostate cancer--do we have a problem with over-detection, overtreatment or both? PMID- 15758700 TI - Is less radical cystectomy better, and how can we be sure? PMID- 15758701 TI - Tolterodine for the treatment of daytime incontinence in children. PMID- 15758702 TI - The burden of urologic diseases in America. PMID- 15758703 TI - Luteinizing hormone-releasing hormone agonist effects on skeletal muscle: how hormonal therapy in prostate cancer affects muscular strength. AB - PURPOSE: Since the discovery of Huggins in 1941 demonstrating the androgen dependence of prostate cancer cells, the use of pharmacological therapies to decrease systemic androgen concentrations has been one of the main treatment options for prostate cancer. Despite their efficacy luteinizing hormone releasing hormone agonists (LHRHas) have a number of side effects, of which many have not been fully investigated in humans. This review focuses on the effects of LHRHas on skeletal muscle in 3 main areas, namely effects at the androgen receptor, at the neuromuscular junction and on skeletal muscle myofibers. Since prostate cancer is predominantly a disease of elderly individuals, the aging effects of LHRHa therapy on skeletal muscle are magnified and of clinical importance. MATERIALS AND METHODS: A comprehensive MEDLINE search was performed of pertinent studies in the literature relating to the use of LHRHa and skeletal muscle. RESULTS: LHRHas affect 3 primary sites within the skeletal muscle system, namely androgen receptor, the neuromuscular junction and second messenger systems, including insulin-like growth factor-1. All sites have been demonstrated to lead to a decrease in isokinetic exercise strength in large muscle groups. CONCLUSIONS: The musculoskeletal effects of LHRHas for the treatment of prostate cancer should be counteracted via a program of exercise strength training to decrease the morbidity associated with skeletal muscle weakness. PMID- 15758704 TI - Laparoscopic radical prostatectomy. AB - PURPOSE: After the pioneering period when only few teams were performing the procedure, the laparoscopic approach to radical prostatectomy has become widespread with several technical variations. A comprehensive review of the published literature on laparoscopic radical prostatectomy was performed to determine the current state of the art of this surgical innovation in terms of perioperative parameters, functional results and cancer control. MATERIALS AND METHODS: English language, peer reviewed articles published before June 2004 concerning laparoscopic radical prostatectomy were found by MEDLINE query. All articles were analyzed and none were a priori excluded. Conclusions were drawn from series of 50 or more patients. RESULTS: Laparoscopic radical prostatectomy is being performed at multiple centers worldwide using various surgical approaches and technologies. Analysis of perioperative parameters, including surgical blood loss, operative time, complications and convalescence, demonstrated low morbidity and showed a clear trend toward improvement with increased experience. The reported positive surgical margin rates were lower in more recent series. As measured by prostate specific antigen recurrence and disease-free intervals, oncological results and cancer control rates are difficult to ascertain in the immature series published to date. Functional results in terms of postoperative urinary and sexual function appear encouraging. CONCLUSIONS: Overall the current operative, oncological and functional results of laparoscopic radical prostatectomy appear to approximate those of open radical retropubic prostatectomy. These results justify the considerable interest of the urological community in laparoscopy, as evidenced by its widespread application. Nevertheless, longer followup and more mature data are needed definitively to establish laparoscopic radical prostatectomy as an alternative to the retropubic approach. PMID- 15758705 TI - Metabolic consequences and long-term complications of enterocystoplasty in children: a review. AB - PURPOSE: We summarize important metabolic consequences and long-term complications associated with enterocystoplasty with particular emphasis on the pediatric patient with genitourinary abnormalities. MATERIALS AND METHODS: A directed MEDLINE literature review for metabolic and long-term complications following enterocystoplasty was performed. Information gained through the published literature and from our database was reviewed and summarized to provide the reader with a thorough review of the subject. RESULTS: Bowel is not a perfect tissue for substitution or augmentation and its use to treat functionally and structurally compromised bladders is associated with several metabolic consequences and long-term complications. Metabolic acidosis is the most common metabolic abnormality seen. The rates and severity of these complications vary, although they may have a profound impact on patient quality of life after enterocystoplasty. CONCLUSIONS: The metabolic consequences and long-term complications associated with enterocystoplasty are important clinical features of this intervention. Careful consideration should be given to them prior to pursuing enterocystoplasty. PMID- 15758706 TI - Legacy of Edwin Beer: fulguration of papillary bladder tumors. AB - PURPOSE: Edwin Beer introduced the first endoscopic treatment of papillary bladder tumors in 1910. This historical review describes how his landmark discovery paved the way for office cystodiathermy to treat recurrent papillary tumors. MATERIALS AND METHODS: The medical writings of Edwin Beer and all studies pertaining to the treatment of papillary bladder tumors from 1879 to the present were reviewed. RESULTS: Edwin Beer discovered that papillary bladder tumors could be destroyed through the ordinary cystoscope with high frequency current by electro-coagulation. This therapy revolutionized the treatment of bladder tumors. CONCLUSIONS: The legacy of Edwin Beer is that office fulguration of recurrent papillary bladder tumors has now become a practical reality due to recent changes in the histological classification of papillary tumors, better definition of their biological behavior and improved endoscopic equipment. PMID- 15758707 TI - Urethral injuries in the Civil War. AB - PURPOSE: We compiled all cases of urethral injury received in battle during the Civil War to detail their management and determine the outcome of treatment. MATERIALS AND METHODS: Surgeon medical reports of individual cases of urethral injury listed in the Medical and Surgical History of the Civil War, and pension records available in the National Archives and Records Administration were reviewed. RESULTS: A total of 105 cases of urethral injury from gunshot wounds, comprising 7% of all urogenital casualties, were reported during the Civil War between 1861 and 1865. Of them 22 (21%) were fatal and 83 patients (79%) cases survived. Debridement and catheter drainage of urinary extravasation or perineal urethrotomy was initial treatment. The majority of urethral injuries were complicated by troublesome strictures, fistulas, urinary incontinence and erectile impotence. Only 19 patients (23%) recovered fully. CONCLUSIONS: Civil War urethral injuries had devastating long-term consequences. PMID- 15758708 TI - Retroperitoneoscopic radical and partial nephrectomy in the patient with cirrhosis. AB - PURPOSE: In patients with cirrhosis and a renal mass options may be limited by medical disease and the surgical difficulties associated with portal hypertension. We describe a retrospective review of patients with cirrhosis with renal masses who underwent radical or partial nephrectomy through a retroperitoneoscopic approach. MATERIALS AND METHODS: Ten consecutive patients, including 4 men and 6 women, with cirrhosis, of whom 2 had undergone liver transplantation, underwent radical (7) or partial (3) nephrectomy for a total of 5 right and 5 left renal neoplasms via the retroperitoneoscopic approach at our institution from March 2002 to February 2004. Recovery data were prospectively obtained and other information was gathered retrospectively from the medical record. RESULTS: Average patient age was 58 years and average American Society of Anesthesiology score was 2.8. Average renal tumor size for radical and partial nephrectomy was 4.6 (range 2.9 to 7) and 1.8 cm (range 1.3 to 2.3), respectively. Operative time was 140 to 315 minutes (median 172) and estimated blood loss was 100 to 5,000 ml (median 225). One patient required open conversion due to hemorrhage from left portosystemic venous communications. Mean postoperative hospitalization was 1.5 days (range 1 to 6). CONCLUSIONS: Although retroperitoneoscopic surgery avoids many surgical dangers associated with portal hypertension and it is our preferred approach to renal surgery in patients with cirrhosis, significant portosystemic venous communications exist in the retroperitoneum, especially on the left side, and they still lead to substantial blood loss in some patients. PMID- 15758709 TI - Assessment of surgical margins in renal cell carcinoma after nephron sparing: a comparative study: laparoscopy vs open surgery. AB - PURPOSE: We compared the status of the peritumoral parenchyma after open and laparoscopic nephron sparing surgery for renal cell carcinoma. MATERIALS AND METHODS: The records of 64 consecutive patients who underwent nephron sparing surgery for renal cell carcinoma of 4 cm or less were reviewed retrospectively. Patients in group 1 underwent open retroperitoneal surgery (1998 to 2000) and patients in group 2 underwent laparoscopic (transperitoneal or retro peritoneal) surgery (2001 to March 2004). A single pathologist was employed to analyze the specimens, and comparative analysis included examination of tumor size, weight, histological cell type, intraoperative histological biopsies and margin status. RESULTS: The 2 groups were comparable in terms of clinical data, and mean lesion size was 31.4 mm in group 1 and 32 mm in group 2. Positive margins were found in 1 of 30 patients in group 1 and in 1 of 34 in group 2 (p = 0.9). An analysis of margins was performed by taking measurements at the minimum and maximum points of the section. The minimum mean measurement was 2 mm in group 1 and 2.08 mm in group 2 (p = 0.75). The maximum mean measurement was 4.56 mm in group 1 and 5.2 mm in group 2 (p = 0.09). The difference between minimum and maximum margin thickness was 2.56 mm in group 1 and 3.16 mm in group 2 (p = 0.04). Mean followup for group 1 was 50 months (range 30 to 72) and 16 months (range 2 to 35) for group 2. One local recurrence was recorded in group 1 and treated with radical nephrectomy, while no recurrence was recorded in group 2. CONCLUSIONS: In this study we further confirmed the efficiency of resectioning lesions using laparoscopy. In our experience there is no difference between the 2 procedures in terms of efficient surgical margins. However, despite these encouraging results it is necessary to obtain more extensive followup data, which will allow us to be more specific in reporting on laparoscopic margin quality. PMID- 15758710 TI - Intermediate followup of hand assisted laparoscopic nephroureterectomy for urothelial carcinoma: factors associated with outcomes. AB - PURPOSE: We report our experience with hand assisted laparoscopic (HALS) nephroureterectomy and describe the associations of preoperative, operative and pathological factors with outcome. MATERIALS AND METHODS: HALS nephroureterectomy was performed in 54 consecutive patients using modified transurethral resection of the ureteral orifice (TURUO) or a 1 port transvesical endoscopic cuff technique for the distal ureter in all except 8. Data were collected prospectively and retrospectively, and followup was distinguished for bladder, contralateral upper tract and nonurothelial (local recurrence and distant metastases) sites. RESULTS: The endoscopic cuff was associated with significantly shorter mean operative time than the transurethral resection of the ureteral orifice method (234 vs 295 minutes, p = 0.002) but the comparison was confounded by the effect of experience. With 28% of patients having stage II or greater tumors and 49% having high grade bladder disease, contralateral upper tract and nonurothelial recurrences developed in 55%, 11% and 25% of evaluable patients at a median followup of 25.1, 24.4 and 24.9 months, respectively, in those without recurrence. At a median followup of 25.0 months cancer specific survival was 94%, 86% and 80% at 1 to 3 years, respectively. Three-year cancer specific survival was 100% in patents with grade 1 or 2, or stage 0 or I tumors but only 57% and 36% in patients with grade 3 and stage II or IV tumors, respectively. CONCLUSIONS: HALS nephroureterectomy is associated with 3-year outcomes that are strongly associated with stage and grade. We prefer the endoscopic cuff method for the distal ureter because it is performed after nephrectomy, does not require patient repositioning and is expedient. PMID- 15758711 TI - A surveillance schedule for G1Ta bladder cancer allowing efficient use of check cystoscopy and safe discharge at 5 years based on a 25-year prospective database. AB - PURPOSE: In the absence of clear evidence, surveillance of low grade superficial bladder cancer by regular check cystoscopy may continue unnecessarily, or discharge from followup may occur empirically. We review the followup during a prospective 25-year period of patients presenting with G1Ta bladder cancer, and it is this analysis on which we base a safe schedule for discharge. MATERIALS AND METHODS: A prospectively kept, computerized record of bladder cancers diagnosed between 1978 and 1985 and subsequently followed up at the Western General Hospital, Edinburgh was reviewed. RESULTS: A total of 115 patients with G1Ta disease were followed for a mean of 19.4 years. Tumor status at 3 months was the strongest prognostic factor for recurrence. Although the absence of tumor at 1 year was also a favorable prognostic sign, it was not for 5 years that the situation entirely stabilized (recurrence developed in 8 of 66 such patients between 1 and 5 years). Of those who did not have recurrence in 5 years, 98.3% patients remained tumor-free for 20 years. In contrast in those with recurrence at 3 months the recurrence rate was much higher. Overall 12% of patients experienced progression, mostly in year 1. None of the 8 who had their first recurrence after year 1 had disease progression. CONCLUSIONS: Patients with G1Ta disease who are free of recurrence for 5 years after presentation can be safely discharged. We propose to alter the regime for patients with no recurrence in year 1 and discharge them at 5 years. PMID- 15758712 TI - Previously unidentified changes in renal cell carcinoma gene expression identified by parametric analysis of microarray data. PMID- 15758713 TI - Solid Renal Tumor Severity in von Hippel Lindau Disease is Related to Germline Deletion Length and Location. PMID- 15758714 TI - Identification of markers for the selection of patients undergoing renal cell carcinoma-specific immunotherapy. PMID- 15758715 TI - Percutaneous CT-Guided Radiofrequency Ablation of Renal Neoplasms: Factors Influencing Success. PMID- 15758716 TI - Differential protein profiling in renal-cell carcinoma. PMID- 15758717 TI - Renal Masses: Quantitative Analysis of Enhancement With Signal Intensity Measurements Versus Qualitative Analysis of Enhancement With Image Subtraction for Diagnosing Malignancy at MR Imaging. PMID- 15758718 TI - Delays in cancer detection using 2 and 4-year screening intervals for prostate cancer screening with initial prostate specific antigen less than 2 ng/ml. AB - PURPOSE: The American Urological Association and American Cancer Society advocate annual screening with serum prostate specific antigen (PSA) and digital rectal examination starting at age 50 years in the general population and earlier in men at high risk. Some groups have suggested that screening at 2 or 4-year intervals may be sufficient in men with initial PSA 2 ng/ml or less. We reviewed the records of men enrolled in a PSA and digital rectal examination based prostate cancer screening study to determine the extent to which the diagnosis of cancer would have been delayed using a 2 or 4-year screening interval. MATERIALS AND METHODS: We evaluated 18,140 volunteers in a prostate cancer screening study in whom PSA was less than 2 ng/ml at initial screening and who were screened at 6 month to 1-year intervals for up to 8 years. We evaluated the cancers detected in these intervals to determine the possible delay in cancer diagnosis that would occur using prolonged screening intervals. We report the overall cancer detection rate, clinical and pathological tumor stage, and Gleason grade of the cancers detected. RESULTS: Excluding 70 men in whom prostate cancer was detected at initial evaluation 2.0% had prostate cancer detected during the next 8 years (mean 21.6 cancers per 6 months, median 20, range 12 to 33). Using a hypothetical 2-year screening interval cancer detection 62% would have been delayed by 4 to 20 months. Using a hypothetical 4-year screening interval cancer detection would have been delayed in 77% of men by 4 to 44 months. Of the tumors detected 100% were clinically localized, 77% were pathologically organ confined and 29% had a Gleason score of 7 or greater. CONCLUSIONS: The 2 or 4-year PSA screening interval in men with initial serum PSA less than 2 ng/ml would result in substantial delays in prostate cancer detection. To our knowledge the extent to which these delays would affect treatment outcomes is undetermined. PMID- 15758719 TI - The extent of lymphadenectomy for pTXNO prostate cancer does not affect prostate cancer outcome in the prostate specific antigen era. AB - PURPOSE: Recent data suggest that extended lymph node dissection in prostate cancer may be necessary for accurate staging. With limited lymph node dissection apparently node negative cases might be under staged. We determined the impact that the number of lymph nodes removed at radical retropubic prostatectomy (RRP) has on cancer progression and cause specific survival in pTXNO cases. MATERIALS AND METHODS: We reviewed the RRP prostate cancer database on 7,036 patients with clinical T1 to T3 disease, no adjuvant therapy and node negative disease in the prostate specific antigen (PSA) era from 1987 to 2000. Factors evaluated were the number of lymph nodes obtained at RRP, preoperative PSA, clinical and pathological stage and grade, margin status, year of surgery and specific surgeon for 5 surgeons who operated throughout the period and performed more than 500 RRPs. Cox analysis was done to determine the RR of progression (PSA or systemic) and prostate cancer death for the number of lymph nodes excised. RESULTS: Median patient age was 65 years and median preoperative PSA was 6.6 ng/ml. At pathological evaluation 5,379 tumors (77%) were organ confined, 4,491 (65%) were Gleason score 5 to 6 and 2,027 (29%) were Gleason score 7 to 10. The median number of nodes obtained significantly decreased from 14 in 1987 to 1989 to 5 in 1999 to 2000 (p <0.001). Ten years after RRP Kaplan-Meier estimates were 63% of cases free of PSA progression, 95% free of systemic progression and 98% free of prostate cancer related death. Median followup was 5.9 years. After adjusting for pathological factors (PSA, grade, stage, margin status and surgical date) the number of lymph nodes obtained at lymphadenectomy was not significantly associated with PSA progression (for each additional node (RR 0.99, 95% CI 0.98 to 1.02, p = 0.90), systemic progression (RR 0.99, 95% CI 0.96 to 1.03, p = 0.68) or cause specific survival (RR 1.01, 95% CI 0.96 to 1.06, p = 0.75). CONCLUSIONS: The extent of lymphadenectomy does not appear to affect prostate cancer outcome in lymph node negative cases. This includes patients with high preoperative PSA, high pathological grade and extracapsular disease. These results suggest that under staging is not present in apparently node negative cases with limited lymphadenectomy and, even if present, its impact on outcome is likely to be negligible. PMID- 15758720 TI - Ability of 2 pretreatment risk assessment methods to predict prostate cancer recurrence after radical prostatectomy: data from CaPSURE. AB - PURPOSE: Two methods widely used to predict the risk of treatment failure after radical prostatectomy for localized prostate cancer are the 3 level D'Amico risk classification and the Kattan nomogram. Although they have been previously validated, to our knowledge they have not been compared in a community based cohort. We tested the 2 instruments in the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) database, a national registry of patients with prostate cancer, to assess their accuracy in a community based cohort. MATERIALS AND METHODS: Men were invited to join CaPSURE from 33 American urology practices, of which 30 were community based. A total of 1,701 men with localized prostate cancer (T1-3a) were treated with radical prostatectomy between 1989 and 2000. Patients who received neoadjuvant or adjuvant therapy were excluded. Recurrence was defined as 2 or more consecutive prostate specific antigen measurements of 0.2 ng/ml or greater, or a second treatment greater than 6 months after surgery. Freedom from progression (FFP) was based on life table estimates and Kaplan-Meier curves. Risk groups were compared using a Cox proportional hazards model and ANOVA. RESULTS: Based on the D'Amico classification 671 cases (39%) were classified as low risk, 446 (26%) were intermediate risk and 584 (34%) were high risk. Five-year FFP was 78%, 63% and 60% in the low, intermediate and high risk groups (HR 1.00, 1.87 and 2.32 respectively, p <0.0001). Mean 5-year FFP predicted by the Kattan nomogram in the same risk groups was 91%, 74% and 69%, respectively. Outcomes in the low risk group were tightly grouped about the mean but there was considerable dispersion of outcomes in the intermediate (30% to 98% FFP) and high (17% to 98%) risk groups. CONCLUSIONS: Stratifying patients in CaPSURE into low, intermediate and high risk categories for disease as described by D'Amico or applying the Kattan nomogram resulted in statistically significant differences in predicted 5-year FFP. However, there was considerable overlap of outcomes between the intermediate and high risk groups. This analysis suggests that simply estimating disease recurrence by stratifying patients into low, intermediate and high risk groups may not provide sufficient information for predicting outcomes among individuals. PMID- 15758721 TI - The impact of obesity on health related quality of life before and after radical prostatectomy (data from CaPSURE). AB - PURPOSE: Health related quality of life (HRQOL) is an important measure of outcomes among patients with prostate cancer due to disease related and treatment related effects on physical and emotional health. We determined if there are differences in the HRQOL of obese men at diagnosis and after radical prostatectomy compared to the HRQOL of men with normal body mass index (BMI). MATERIALS AND METHODS: Data were abstracted from Cancer of the Prostate Strategic Urological Research Endeavor (CaPSURE), a disease registry of 10,018 men with prostate cancer. A total of 1,884 men were included in study who were treated with radical prostatectomy between 1989 and 2002, had BMI information available and had completed 1 initial HRQOL questionnaire. Of these men 672 who completed at least 2 followup questionnaires were assessed further. RESULTS: The BMI (kg/m) distributions were 24% normal (less than 24.9 kg/m), 56% overweight (25 to 29.9), 16% obese (30 to 34.9) and 4% very obese (greater than 35 kg/m). Higher BMI was associated with worse physical function, bodily pain, general health, vitality and role physical, but better bowel bother at diagnosis independent of race. Higher BMI was also associated with worse HRQOL after radical prostatectomy for physical function, general health and vitality, but better bowel bother. HRQOL differences between BMI groups were similar among times for all measured variables. Compared to the normal group, the higher BMI groups had similar HRQOL after radical prostatectomy. CONCLUSIONS: In the majority of domains men with higher BMI had lower HRQOL at diagnosis than men of normal BMI. Obese men have a similar recovery pattern of HRQOL after radical prostatectomy, with minimal additive long-term impairment in HRQOL relative to men of normal weight. PMID- 15758722 TI - Intraoperative electrophysiological confirmation of urinary continence after radical prostatectomy. AB - PURPOSE: To determine the actual effect of nerve sparing radical retropubic prostatectomy (RP) on postoperative urinary continence we used intraoperative electrophysiological testing to confirm functional preservation of the neurovascular bundle (NVB). MATERIALS AND METHODS: A total of 85 patients undergoing RP for localized prostate cancer were studied. During RP NVB preservation was assessed macroanatomically. Electrophysiological testing was then performed to confirm NVB preservation. The NVB was electrostimulated and responses were observed by monitoring intracavernous or intraurethral pressure changes. All patients were classified into 3 groups according to the degree of nerve sparing, that is a bilateral nerve sparing group, a unilateral nerve sparing group and a nonnerve sparing group, based on macroanatomical assessment as well as on electrophysiological assessment. Postoperative continence in each group was then determined. Urinary continence at baseline, and 3 and 6 months postoperatively was studied using a self-administered questionnaire. RESULTS: With electrophysiological assessment 20.6% of macroanatomically determined NVB preservations were reclassified. Analysis of the data on groups classified accurately by electrophysiological testing showed that the bilateral nerve sparing group maintained postoperative urinary function significantly more than the unilateral nerve sparing and nonnerve sparing groups. However, when only macroanatomical assessment was considered, no significant difference among the groups was found in urinary function. CONCLUSIONS: Electrophysiological assessment revealed that bilateral NVB preservation contributes to early recovery of urinary continence after RP. Thus, intraoperative electrophysiological assessment is useful for predicting postoperative quality of life. PMID- 15758723 TI - Anastomotic contracture and incontinence after radical prostatectomy: a graded approach to management. AB - PURPOSE: We present a heterogeneous group of men presenting with varying degrees of anastomotic contracture (AC) and associated stress urinary incontinence (SUI) following radical prostatectomy. It is particularly important that AC should be resolved before artificial urinary sphincter (AUS) implantation, because instrumentation through the AUS can risk erosion. MATERIALS AND METHODS: The records of 54 consecutive men who were referred for the management of AC and associated SUI were reviewed. Patient treatment and outcomes were stratified according to their unique characteristics. RESULTS: A total of 54 patients underwent radical prostatectomy alone (48), or in combination with radiation therapy (7) or cryotherapy (1). In group 1, 35 patients had previously undiscovered AC, or 1 or more prior contracture incisions (CIs) with SUI. CI and AUS were performed simultaneously in 33 patients and sequentially in 2. In group 2, 7 patients with intractable AC following multiple CIs/dilations and self calibration, or an indwelling urethral or suprapubic catheter underwent simultaneous (3) or sequential (2) CI/AUS or CI only (2). Five patients required temporary self-calibration. In group 3, in 12 patients with total outlet obliteration recanalization was accomplished with combined antegrade/retrograde endoscopy and CI. Ten patients had re-obliteration, of whom 1 underwent suprapubic diversion and 9 underwent repeat recanalization with placement of a UroLume stent (American Medical Systems, Minnetonka, Minnesota) across the anastomosis. Eight patients underwent artificial urinary sphincter (AUS) placement 4 to 6 weeks later and 1 awaits an AUS. Of those implanted with an AUS 2 required repeat endoscopic procedures because of recurrent but manageable stent ingrowth. CONCLUSIONS: Most ACs are treated successfully with simultaneous, aggressive CI/AUS. A history of many CIs or long, dense contractures suggest the need for staged management. In those with obliterated outlets we prefer to reestablish patency and if rapid recurrence develops, we place a UroLume stent. Regardless of a history of radiation therapy, continence is restored with an AUS. PMID- 15758724 TI - Does fibrin sealant decrease immediate urinary leakage following radical retropubic prostatectomy? AB - PURPOSE: We determined the effectiveness of fibrin sealant in decreasing postoperative urinary leakage following radical retropubic prostatectomy performed by 1 surgeon at Washington Hospital Center. MATERIALS AND METHODS: Between April and November 2003 our group treated 32 consecutive patients with prostate cancer with radical retropubic prostatectomy. The first 16 patients (control) underwent the Walsh described technique and the second group of 16 patients had an additional application of fibrin sealant around the urethro vesical anastomosis. Postoperative drain output was measured every 8 hours. The results of the 2 groups were compared. RESULTS: The Blake drain was removed after 4 nursing shifts (times 1 through 4) in 81% (13 of 16) of the control group and in 100% (16 of 16) of the fibrin sealant group. The fibrin sealant group had significantly less drainage output overall compared with the control group (p = 0.005). The drainage output from each group decreased with time at a significant rate independent of each other (p <0.001), and there was a larger difference (p = 0.04) in output between groups at times 1 and 2 compared with times 3 and 4. There was no relationship between the amount of urinary drainage and drain output. There was no immediate morbidity associated with the use of fibrin sealant. CONCLUSIONS: The application of fibrin sealant to the urethro vesical anastomosis during radical retropubic prostatectomy does decrease postoperative drain output. With earlier drain removal, patients would benefit from less discomfort and from skilled nursing requirements. In select patients early drain removal could accelerate discharge home. PMID- 15758725 TI - Contemporary survival results and the role of radiation therapy in patients with node negative seminal vesicle invasion following radical prostatectomy. AB - PURPOSE: Seminal vesicle invasion (SVI) in a radical prostatectomy (RRP) specimen is associated with a guarded prognosis. We evaluated patients with SVI treated in the pre-prostate specific antigen (PSA) (1983 to 1991) and PSA (1992 to 2003) eras. MATERIALS AND METHODS: Of patients with prostate cancer treated with RRP from January 1983 through March 2002, 220 with SVI were evaluated, including 67 in the pre-PSA era and 153 in the PSA era. Postoperative PSA greater than 0.2 ng/ml was considered biochemical evidence of cancer progression. Survival rates were compared using Kaplan-Meier estimates to calculate progression-free, cancer specific and all cause survival. Multivariate Cox proportional hazard models were used to correlate variables with disease progression. RESULTS: The incidence of SVI in the PSA era was lower than in the pre-PSA era (6.0% vs 10.2%, p = 0.001). To date 124 patients (56%) have had evidence of cancer progression. The 4 and 7 year progression-free, cancer specific and all cause survival rates were significantly higher in men with SVI in the PSA era (p = 0.02). PSA at diagnosis, cancerous surgical margins and higher Gleason score were significantly associated with progression. Neither adjuvant nor salvage radiotherapy appeared to confer a significant progression-free survival benefit. CONCLUSIONS: The incidence of SVI has decreased in the PSA era. Progression-free, cancer specific and all cause survival rates following RRP in patients with SVI have improved in the PSA era. This may reflect earlier detection in this pathological tumor stage and more favorable prognostic factors associated with PSA screening. Adjuvant radiotherapy does not appear to confer any therapeutic benefit. Salvage radiotherapy can lead to durable PSA regressions in a small percent of men, although no long-term survival advantage can be proved. PMID- 15758727 TI - Radiotherapy for basaloid carcinoma of the penis: a case report. PMID- 15758726 TI - Salvage surgery for radiorecurrent prostate cancer: contemporary outcomes. AB - PURPOSE: We present a 30-year experience with performing salvage surgery in patients with persistent prostate cancer (PCA) after definitive radiotherapy. MATERIALS AND METHODS: Patients with biopsy proven PCA following definitive radiotherapy who underwent salvage surgery were identified retrospectively (1967 to 2000). Prostate specimens were evaluated by a single pathologist. Progression free survival (PFS) and cancer specific survival (CSS) estimates were made with multivariate analysis of outcome predictors. Complications were reviewed. RESULTS: Sufficient information was available on 199 patients, including 138 with retropubic prostatectomy (RP) and 61 with cystoprostatectomies (CP). Median followup was 7.0 years. Rectal injury rates (5% for RP and 10% for CP) remained stable, while transfused units of blood decreased. Urinary extravasation (15% of cases) and bladder neck contracture (22%) were the most common complications. Urinary continence (0 pads) improved from 43% to 56% of patients (p = 0.17) with an additional 20% requiring 1 or fewer pad daily. Overall 10-year CSS in all patients undergoing salvage surgery was 65%. Patients undergoing RP fared better than those needing CP (10-year CSS 77% vs 38%, p <0.001 and median PFS 8.7 vs. 4.4 years, p <0.001). Tumor ploidy, percent 4/5 Gleason grade and pathological stage were strong predictors of outcome, while margin status and preoperative prostate specific antigen had minimal predictive strength. CONCLUSIONS: Significant PFS and CSS can be expected following salvage surgery for radioresistant PCA. Several pathological features of the removed prostate are predictive of survival. The surgical risks of salvage surgery are now defined. Morbidity rates, including continence, have moderately improved with time. PMID- 15758728 TI - Urethral tumor recurrence following cystectomy and urinary diversion: clinical and pathological characteristics in 768 male patients. AB - PURPOSE: We evaluated the incidence and risks of urethral recurrence following radical cystectomy and urinary diversion in men with transitional cell carcinoma of the bladder. MATERIAL AND METHODS: Clinical and pathological results were evaluated in 768 consecutive male patients undergoing radical cystectomy with intent to cure for bladder cancer with a median followup 13 years, including 397 (51%) who underwent orthotopic urinary diversion with a median followup of 10 years and 371 (49%) who underwent cutaneous urinary diversion with a median followup of 19 years. Demographically and clinically these 2 groups were well matched with the only exception being longer median followup in the cutaneous group (p <0.001). Urethral recurrence was analyzed by univariate and multivariable analysis according to carcinoma in situ, tumor multifocality, pathological characteristics (tumor grade, stage and subgroup), the presence and extent of prostate tumor involvement (superficial vs stromal invasion) and the form of urinary diversion (cutaneous vs orthotopic). RESULTS: A total of 45 patients (6%) had urethral recurrence at a median of 2 years (range 0.2 to 13.6), including 16 (4%) with an orthotopic and 29 (8%) with a cutaneous form of urinary diversion. Carcinoma in situ and tumor multifocality were not significantly associated with an increased risk of urethral recurrence (p = 0.07 and 0.06, respectively). The presence of any (superficial and/or stromal invasion) prostatic tumor involvement was identified in 129 patients (17%). Prostate tumor involvement was associated with a significantly increased risk of urethral recurrence (p = 0.01). The estimated 5-year chance of urethral recurrence was 5% without any prostate involvement, increasing to 12% and 18% with superficial and invasive prostate involvement, respectively. Patients undergoing orthotopic diversion demonstrated a significantly lower risk of urethral recurrence compared with those undergoing cutaneous urinary diversion (p = 0.02). Patients without any prostate tumor involvement and orthotopic diversion (lowest risk group) demonstrated an estimated 4% year chance of urethral recurrence compared with a 24% chance in those with invasive prostate involvement undergoing cutaneous diversion (highest risk group). On multivariate analysis any prostate involvement (superficial and/or invasive) and urinary diversion form remained independent and significant predictors of urethral recurrence (p = 0.035 and 0.01, respectively). CONCLUSIONS: At long-term followup urethral tumor recurrence occurs in approximately 7% of men following cystectomy for bladder transitional cell carcinoma. Involvement of the prostate with tumor and the form of urinary diversion were significant and independent risk factors for urethral tumor recurrence. Patients undergoing orthotopic diversion have a lower incidence of urethral recurrence compared with those undergoing cutaneous diversion. Although prostate tumor involvement is a risk factor for urethral recurrence, it should not preclude orthotopic diversion, provided that intraoperative frozen section analysis of the urethral margin is without evidence of tumor. PMID- 15758729 TI - Hedgehog signalling in prostate regeneration, neoplasia and metastasis. PMID- 15758731 TI - Alcohol consumption and risk of prostate cancer in middle-aged men. PMID- 15758730 TI - Prostate cancer screening and mortality: a case-control study (United States). PMID- 15758732 TI - Low p21Waf1/Cip1 Protein Level Sensitizes Testicular Germ Cell Tumor Cells to Fas Mediated Apoptosis. PMID- 15758733 TI - Delayed intraoperative hydration limits blood loss during radical retropubic prostatectomy. PMID- 15758734 TI - Histologic Grading of Noninvasive Papillary Urothelial Tumors. Validation of the 1998 WHO/ISUP System by Immunophenotyping and Follow-Up. PMID- 15758735 TI - Histogenesis of nonurothelial carcinomas of the urinary bladder from pre-existent transitional cell carcinomas. A histopathological and immunohistochemical study. PMID- 15758736 TI - Antibiotic prophylaxis for transurethral prostatic resection in men with preoperative urine containing less than 100,000 bacteria per ml: a systematic review. AB - PURPOSE: We determined whether antibiotic prophylaxis can reduce the risk of postoperative infective complications in men undergoing transurethral resection of the prostate (TURP) who have preoperative urine with less than 100,000 bacteria per ml. MATERIALS AND METHODS: MEDLINE, EMBASE (Elsevier B.V., Amsterdam, The Netherlands) and the Cochrane Library were searched for randomized and quasi-randomized controlled trials that compared the effects of antibiotic prophylaxis with placebo or active controls for men undergoing TURP with preoperative sterile urine. Two reviewers independently extracted patient characteristic and outcomes data based on a prospectively developed protocol. RESULTS: A total of 28 trials, 10 placebo controlled and 18 no treatment controlled, involving 4,694 patients, met the inclusion criteria. The mean age of the subjects was 69 years and the majority underwent TURP for prostatic hyperplasia (85%). Antibiotic prophylaxis was significantly more effective than placebo in reducing postoperative TURP complications. The risk differences for post-TURP bacteriuria, high degree fever, bacteremia and use of additional antibiotic treatment were -0.17 (95% CI 0.20, -0.15), -0.11 (-0.15, -0.06), -0.02 (-0.04, 0.00) and -0.20 (-0.28, -0.11), respectively. The results were observed consistently across all classes of antibiotics assessed. There was no difference in the duration of postoperative catheterization or hospitalization. Adverse events were rare, generally mild, and included allergic reactions, pyrexia and abdominal complaints. CONCLUSIONS: Prophylactic antibiotics decrease the incidence of post-TURP bacteriuria, high fever, bacteremia and additional antibiotic treatment. Additional research should evaluate the optimal antibiotic regimen, and whether the cost and possibility of the development of resistant strains of organisms justify the routine use of prophylactic antibiotics. PMID- 15758737 TI - Abnormal urinary potassium metabolism in patients with interstitial cystitis. AB - PURPOSE: If most patients with interstitial cystitis (IC) have epithelial leakage allowing urinary K to penetrate the interstitium and provoke symptoms, urinary K should be lower in untreated patients than in healthy subjects and it should increase with successful heparinoid treatment. This study tested these hypotheses. MATERIALS AND METHODS: Na, K and creatinine (Cr) were determined in spot urine samples from new, symptomatic, untreated patients with IC meeting all National Institute of Diabetes and Digestive and Kidney Diseases clinical diagnostic criteria, returning patients with IC reporting 50% or greater symptom improvement after 4 or greater months of oral heparinoid therapy and control subjects, and in 24-hour urine samples from new untreated patients and controls. RESULTS: In spot urine specimens of 37 new patients with IC K-to-Cr ratios were significantly lower than in 18 controls (0.51 vs 0.88 mg/mg Cr, p = 0.001). A total of 50 successfully treated patients with IC had significantly higher K-to Cr ratios than those in 37 new patients (0.66 vs 0.51 mg/mg Cr, p = 0.025). Na-to Cr ratios in the 3 groups were not significantly different. In 24-hour urine specimens 30 new patients had lower average K (31.0 vs 46.2 mEq/l, p = 0.01) and lower K-to-Cr ratios (0.43 vs 0.52 mg K/mg Cr, p = 0.01) than in 47 controls, while Na was not significantly different. CONCLUSIONS: Our finding of lower urinary K in new, untreated patients supports the concept of abnormal epithelial permeability and K absorption in IC. Higher urinary K in successfully treated vs untreated patients may reflect decreasing urinary K absorption due to mucosal repair and a resulting decrease in epithelial permeability. K/mg Cr appears accurate for normalizing urinary K. PMID- 15758738 TI - A randomized controlled trial of intravesical bacillus calmette-guerin for treatment refractory interstitial cystitis. AB - PURPOSE: We compared intravesical bacillus Calmette-Guerin (BCG) to placebo instillations in patients with treatment refractory interstitial cystitis (IC). MATERIALS AND METHODS: Subjects who met the National Institutes of Health National Institute for Diabetes and Digestive and Kidney Diseases criteria for IC, and reported at least moderate pain and frequency for a minimum of 6 months before study entry, were randomized to 6 weekly double-blinded intravesical instillations of either BCG or placebo, and then followed for a total of 34 weeks. The primary outcome was a patient reported global response assessment at week 34, supplemented with medications for IC during weeks 31 to 34. Secondary outcomes included a 24-hour voiding diary, pain, urgency, validated IC symptom indexes and adverse events. The target sample size was 260 participants, designed to detect a difference in response rates between placebo and BCG of 30% and 50%, respectively. RESULTS: A total of 265 participants were randomized and 17 (6%) patients withdrew from study. The response rates for the primary outcome were 12% for placebo and 21% for BCG (p = 0.062). Small improvements were observed for all secondary outcomes, some more so with BCG, but these differences were of borderline statistical significance. Although a large number of adverse events were reported in the BCG arm, there was no statistically significant difference between the treatment arms in overall adverse event rates. CONCLUSIONS: Although the BCG safety profile was acceptable, the response rate for the primary outcome was low. Effective medical treatment for patients with moderate to severe interstitial cystitis remains elusive. PMID- 15758740 TI - Effect of circumcision on urinary tract infection after successful antireflux surgery. PMID- 15758739 TI - Cost-effectiveness analysis of bacterial endocarditis prophylaxis for febrile children who have cardiac lesions and undergo urinary catheterization in the emergency department. PMID- 15758741 TI - Extended-interval aminoglycoside administration for children: a meta-analysis. PMID- 15758743 TI - Pain after percutaneous nephrolithotomy: impact of nephrostomy tube size. PMID- 15758742 TI - Percutaneous nephrolithotomy for caliceal diverticular calculi: a novel single stage approach. AB - PURPOSE: Current percutaneous treatment of symptomatic caliceal diverticular calculi involves renal access, stone removal, dilation of the diverticular communication, fulguration of the cavity and placement of a nephrostomy tube. We reviewed the outcomes of patients undergoing a novel single stage percutaneous nephrolithotomy technique for radiopaque caliceal diverticular stones that eliminates ureteral catheterization and entry into the renal collecting system. MATERIALS AND METHODS: A total of 21 patients (8 male and 13 female including 1 bilateral) with a mean age of 42.4 years underwent percutaneous nephrolithotomy for caliceal diverticular stones from February 2001 to May 2003. Of the diverticula 12 were upper pole, 4 were interpolar and 6 were lower pole. Infracostal access was established by the urologist directly onto the radiopaque stones without the aid of a ureteral catheter. After balloon tract dilation a 30Fr Amplatz sheath was placed and following stone removal the diverticulum was fulgurated. The infundibulum was neither cannulated nor dilated. A 20Fr red rubber catheter or an 8.5Fr Cope loop was placed into the diverticulum. Stone free status was assessed by noncontrast computerized tomography on postoperative day 1 (POD1). The drainage tube was removed if there was no urine drainage and the kidney was stone-free. Excretory urography was performed at 3 months to evaluate diverticular resolution. RESULTS: Of 21 patients 20 were discharged home tubeless on POD1 and 18 of 21 (85.7%) renal units were stone- free on POD1 noncontrast computerized tomography. Mean operative time was 58.5 minutes and mean stone burden was 138.9 mm. Mean stone diameter was 11.6 mm and mean diverticular diameter was 15.3 mm. Of 22 renal units 16 had followup excretory urography. All diverticula decreased in size and 14 (87.5%) had complete resolution. CONCLUSIONS: In patients with symptomatic radiopaque caliceal diverticular stones, a single stage procedure without the need for ureteral catheterization combined with direct infracostal diverticular puncture allows for a rapid procedure with little morbidity. PMID- 15758744 TI - Percutaneous nephrolithotomy: an update. PMID- 15758745 TI - Laparoscopic partial nephrectomy using holmium laser in a porcine model. PMID- 15758746 TI - Long-term combined treatment with thiazide and potassium citrate in nephrolithiasis does not lead to hypokalemia or hypochloremic metabolic alkalosis. PMID- 15758747 TI - Laparoscopic radical nephrectomy: oncologic efficacy. PMID- 15758748 TI - Traumatic ureteral injuries: a single institution experience validating the American Association for the Surgery of Trauma-Organ Injury Scale grading scale. AB - PURPOSE: Ureteral injuries are uncommon and challenging. In this study we report our institutional experience with ureteral injuries. We evaluated the American Association for the Surgery of Trauma-Organ Injury Scale (AAST-OIS) for ureteral injuries as a predictor of outcomes for complexity of repair, morbidity, mortality and associated injuries. MATERIALS AND METHODS: We performed a retrospective, 120-month study (January 1992 to December 2002) at an urban, level I trauma center. RESULTS: In the 57 patients mean hospital Admission blood pressure +/- SD was 115 +/- 25 mm Hg, mean Revised Trauma Score was 7.38 +/- 0.84 and mean Injury Severity Score was 15 +/- 1.15. The mechanism of injury was penetrating in 55 cases (96.5%), including gunshot wound in 52 (54.5%) and stab wound in 2 (5.5%), and in blunt 2 of motor vehicle accidents (3.5%). The anatomical location was the left side in 33 cases (58%), right side in 23 (40%) and bilateral in 1 (2%). The distribution of injuries was proximal in 15 cases (26%), mid in 21 (37%) and distal in 21 (37%). Associated injuries were present in 56 patients (98%). An intraoperative diagnosis was made in 44 cases (77%). Of the patients 50 (88%) required complex repairs or an adjunct procedure, including a double pigtail stent in 33 (58%), ureteroureterostomy in 20 (35%), ureteroneocystostomy with a psoas hitch in 10 (18%), external diversion in 9 (16%), suprapubic cystostomy in 8 (14%), nephrostomy in 2 (3.5%), nephrectomy in 2 (3.5%) and ligation in 2 (3.5%). Injury grade was I to V in 5 (8%), 8 (13%), 13 (22.8%), 18 (31.6%) and 13 (22.8%) cases, respectively. Overall 51 patients (89%) survived. No deaths were related to ureteral injury. Renal salvage was achieved in 49 of the 51 surviving patients (96%). CONCLUSIONS: Ureteral injuries are uncommon. The complexity of repair and number of associated injuries increase with AAST-OIS injury grade. Mortality increases with AAST-OIS injury grade but it is not related to the ureteral injury. Excellent results can be achieved with complex techniques of primary repair, leading to renal salvage. PMID- 15758749 TI - Primary urethral reconstruction: the cost minimized approach to the bulbous urethral stricture. AB - PURPOSE: Treatment for urethral stricture disease often requires a choice between readily available direct vision internal urethrotomy (DVIU) and highly efficacious but more technically complex open urethral reconstruction. Using the short segment bulbous urethral stricture as a model, we determined which strategy is less costly. MATERIALS AND METHODS: The costs of DVIU and open urethral reconstruction with stricture excision and primary anastomosis for a 2 cm bulbous urethral stricture were compared using a cost minimization decision analysis model. Clinical probability estimates for the DVIU treatment arm were the risk of bleeding, urinary tract infection and the risk of stricture recurrence. Estimates for the primary urethral reconstruction strategy were the risk of wound complications, complications of exaggerated lithotomy and the risk of treatment failure. Direct third party payer costs were determined in 2002 United States dollars. RESULTS: The model predicted that treatment with DVIU was more costly (17,747 dollars per patient) than immediate open urethral reconstruction (16,444 dollars per patient). This yielded an incremental cost savings of $1,304 per patient, favoring urethral reconstruction. Sensitivity analysis revealed that primary treatment with urethroplasty was economically advantageous within the range of clinically relevant events. Treatment with DVIU became more favorable when the long-term risk of stricture recurrence after DVIU was less than 60%. CONCLUSIONS: Treatment for short segment bulbous urethral strictures with primary reconstruction is less costly than treatment with DVIU. From a fiscal standpoint urethral reconstruction should be considered over DVIU in the majority of clinical circumstances. PMID- 15758751 TI - Renovascular injury: an argument for renal preservation. PMID- 15758750 TI - Pelvic fracture-associated urethral injuries in girls: experience with primary repair. PMID- 15758752 TI - Parastomal hernia repair using the lateral approach. PMID- 15758753 TI - Laparoscopic repair of parastomal hernias. Early results. PMID- 15758754 TI - An assessment of the use of a continent catheterizable stoma in female tetraplegics. PMID- 15758755 TI - Increased warning time with darifenacin: a new concept in the management of urinary urgency. AB - PURPOSE: We assessed the effect of darifenacin, an M3 selective receptor antagonist, on the warning time associated with urinary urgency. MATERIALS AND METHODS: In this multicenter, double-blind study subjects with urinary urgency for 6 months or greater and episodes of urgency 4 times or greater daily were randomized to darifenacin controlled release tablets (30 mg once daily) or placebo. Warning time was defined as the time from the first sensation of urgency to voluntary micturition or incontinence. Data were collected using electronic event recorders during 6-hour clinic visits or 3 urge-void cycles, if shorter, at baseline and at treatment end. RESULTS: A total of 72 subjects entered the study and 67 were included in the primary efficacy analysis (darifenacin in 32 and placebo in 35). Darifenacin treatment resulted in a significant increase in mean warning time with a median increase of 4.3 minutes compared with placebo (p = 0.003). Overall 47% of darifenacin treated subjects compared with 20% receiving placebo achieved a 30% increase or greater in mean warning time (OR 5.6, p = 0.009). Median and minimum warning times were also significantly increased following darifenacin treatment vs placebo (p = 0.004 and 0.017, respectively). The median difference in minimum warning time was 1.9 minutes in favor of darifenacin vs placebo. CONCLUSIONS: To our knowledge this is the first study to evaluate change in warning time, which is potentially important to individuals with symptoms associated with overactive bladder. Darifenacin increases mean, median and minimum warning time compared with placebo, allowing subjects more time to reach a toilet and potentially avoiding the embarrassing experience of incontinence. PMID- 15758756 TI - Association between valsalva and cough leak point pressures and pelvic organ prolapse quantification in women with stress incontinence. AB - PURPOSE: Women with urodynamically documented stress urinary incontinence (SUI) and urethral hypermobility may have a higher pelvic organ prolapse quantification (POP-Q) stage according to anterior POP-Q measurements. In this study we determined if POP-Q system anterior components representing the urethrovesical junction (anterior wall point Aa/Ba) and/or POP-Q stage has a relationship with leak point pressure testing. MATERIALS AND METHODS: Of the 1,511 women who underwent video fluoro-urodynamics during 1997 to 2003 at our institution 88 with only evidence of SUI with negative Valsalva leak point pressure and positive cough leak point pressure (CLPP) were selected. RESULTS: Average patient age was 58.6 years (range 32 to 89). Of the 88 women 82 had complete POP-Q examinations available, which revealed stages 0 to III in 21 (25.61%), 20 (24.39%), 40 (48.78%) and 1 (1.22%), respectively. The association between POP-Q stage/components and positive CLPP showed no significant difference in mean positive CLPP among POP-Q stages (p = 0.178) or components (p = 0.42 to 0.97). The test for linear trend was not significant (p = 0.636) for POP-Q stages/components (p = 0.40 to 0.93). No significant difference in volume at which positive CLPP occurred was observed among POP-Q stages (p = 0.283) or components (p = 0.13 to 0.75). The proportion of patients with leakage at 200 cc did not differ significantly among POP-Q stages (p = 0.119) or components (p = 0.15 to 0.60). CONCLUSIONS: Analysis of women with urodynamic evidence of SUI with negative Valsalva leak point pressure and positive CLPP did not show any significant association with components of the POP-Q system or with POP-Q stages. Findings support that POP-Q measurements should not be interpreted as indicators of urethral hypermobility when evaluating women with SUI. PMID- 15758757 TI - Transobturator vaginal tape inside out for the surgical treatment of female stress urinary incontinence: anatomical considerations. AB - PURPOSE: We have recently described a novel surgical technique for female stress urinary incontinence, that is the transobturator vaginal tape inside out, which uses specific instruments for the passage of a synthetic tape from beneath the urethra toward the thigh folds. Herein we report the results of cadaver dissection performed to determine the anatomical trajectory of the tape and its relationships with neighboring neurovascular structures and organs. MATERIALS AND METHODS: Insertion of the transobturator vaginal tape inside out tape was performed by different surgeons in 12 freshly frozen female cadavers according to the standard procedure. The thigh, obturator, perineal and pelvic regions were dissected and tape trajectory was recorded. An additional cadaver was dissected without prior tape placement. RESULTS: The tape was inserted according to a certain consistent path, that is penetration from the suburethral space into a strictly perineal region limited medial and cranial by the levator ani muscle, caudal by the perineal membrane and lateral by the obturator internus muscle. This region corresponded to the most anterior recess of the ischiorectal fossa. The tape then perforated the obturator membrane and muscles, and exited through the skin after traversing adductor muscles and subcutaneous tissue. The tape was coursed away from 1) the dorsal nerve to the clitoris located more superficially below the perineal membrane, 2) the obturator nerve and vessels, and 3) the saphenous and femoral vessels. CONCLUSIONS: These findings strongly suggest that our transobturator technique is highly accurate, reproducible and safe, and it does not require perioperative cystoscopy. PMID- 15758758 TI - Cadaveric prolapse repair with sling: intermediate outcomes with 6 months to 5 years of followup. AB - PURPOSE: We present the prospective, intermediate-term results for cadaveric prolapse repair with sling as combined treatment of stress urinary incontinence and cystocele. MATERIALS AND METHODS: A total of 251 of 295 (85%) patients undergoing cadaveric prolapse repair with sling (CaPS) had at least 6 months of questionnaire and pelvic examination followup. All patients had objectively demonstrated stress urinary incontinence and grade 2 to 4 cystocele before surgery. Followup outcome measures included a validated subjective continence and patient satisfaction questionnaire, SEAPI (stress incontinence, emptying, anatomy, protection, inhibition) scores, pelvic examination for prolapse recurrence and complications, and quality of life scores. RESULTS: The overall patient reported subjective incontinence cured/dry rate (no incontinence episodes of any type) was 114 of 251 (45%), the cured/improved rate (50% improvement or greater) was 192 of 251 (76%) and the failed rate (less than 50% improvement) was 59 of 251 (24%). When considering stress urinary incontinence the cured/dry rate was 141 of 251 (56%), cured/improved rate was 207 of 251 (82%) and failed rate was 44 of 251 (17.5%), with 17 of the 44 (39%) cases having mixed urinary incontinence. Of the 59 failures 33 (56%) occurred after 12 months of followup. Of 153 patients 13 (8.5%) experienced de novo urge incontinence. The symptomatic cystocele recurrence rate was 18 of 251 (7%). There were statistically significant improvements in SEAPI and prolapse quality of life scores. Of 251 patients 200 (80%) were at least 50% satisfied and of 251 193 (77%) stated they would undergo the CaPS procedure again. CONCLUSIONS: With a maximum followup of 5 years in patients undergoing CaPS, we have seen excellent, durable cystocele repair results. While our subjective continence rates have decreased with an increasing number of late failures, we continue to observe significant improvement in SEAPI scores and quality of life with good patient satisfaction and low morbidity. PMID- 15758759 TI - Long-term 5-year followup of the results of the vesica procedure. AB - PURPOSE: We evaluated the long-term results of Vesica (Boston Scientific Corp., Watertown, Massachusetts) percutaneous bladder neck suspension for stress urinary incontinence. MATERIALS AND METHODS: A total of 40 women with urodynamically proven stress urinary incontinence (SUI) underwent Vesica percutaneous bladder neck suspension between 1994 and 1997. Patients were assessed at 6 months, 12 months and 5 years with a simple questionnaire to elicit whether they had experienced any adverse effects, whether they were dry and whether further investigation or a surgical incontinence procedure was offered. RESULTS: Only 1 of the 40 women was lost to long-term followup. Initial results were excellent with 85% of women reporting complete dryness at 6 months. However, wound infections developed in 16% of patients secondary to hematomas in the suprapubic incisions and 10% required a period of intermittent self-catheterization. By 12 months only 46% of women remained dry, although most only reported occasional leakage. At 5 years 69% of patients had recurrent SUI and more than two-thirds of this group (70%) had symptoms severe enough to be offered a further surgical procedure. Patients undergoing subsequent secondary procedures were found to have fraying of the suspensory sutures at the bone anchor. CONCLUSIONS: Initial results of this minimally invasive procedure were excellent and despite the lack of long-term data the technique rapidly came into widespread use. The 5-year outcome shows a 31% continence rate. We no longer advocate this particular form of bladder neck suspension for SUI. PMID- 15758760 TI - Simultaneous augmentation cystoplasty is associated with earlier rather than increased artificial urinary sphincter infection. AB - PURPOSE: While artificial urinary sphincter infection or erosion occurs in 20% of implantations, the risk factors are poorly understood. One of the most contentious factors reported to increase prosthesis infection is simultaneous sphincter implantation and augmentation cystoplasty. In contrast to some reports, to date our results have not shown an increased infective risk with the simultaneous procedure. We reviewed the long-term infective complications of 195 sphincters to investigate for predisposing infective factors and review the role of augmentation cystoplasty. MATERIALS AND METHODS: We performed a retrospective case note review of 144 patients with a median followup of 112 months. Augmentation cystoplasty performed in 86 patients (60%) and was simultaneous in 56. All patients were reviewed within the last year or followed until death or sphincter failure. Patient, surgical and treatment factors were statistically analyzed for associations with prosthesis infection. RESULTS: A total of 108 sphincters failed from infection (25%), tissue atrophy (5%) or mechanical reasons (25%). The overall infective failure rate was similar in patients who underwent simultaneous augmentation (30%) compared with the other patients (23%), although there was a statistically significant difference within the first 3 postoperative years (log rank p = 0.009). While no other variables were significantly associated with sphincter infection, intermittent self-catheterization did not increase sphincter infection and females appeared to have more prosthesis infections. CONCLUSIONS: Our results suggest that, while simultaneous augmentation cystoplasty and artificial urinary sphincter implantation lead to an initial increase in prosthesis infection, this difference disappears after 3 years. PMID- 15758761 TI - Outcomes following revisions and secondary implantation of the artificial urinary sphincter. AB - PURPOSE: Durable success with the artificial urinary sphincter (AUS) is common but device revision and replacement are often needed for various reasons. We examined indications and outcomes following these secondary procedures with comparisons to outcomes after primary procedures. MATERIALS AND METHODS: The medical records of all patients undergoing primary and secondary bulbar urethral AUS implantation and revision from January 1990 to September 2002 were reviewed for various demographic and surgical variables. Female patients and males with bladder neck cuffs were excluded from study. RESULTS: Of 554 men undergoing AUS implantation or revision 119 (21.4%) underwent a total of 159 secondary procedures. Reasons for revision were mechanical failure in 31 cases (25.2%) and nonmechanical failure in 88 (73.9%). The latter included recurrent incontinence due to urethral atrophy in 63 cases (52.9%) and erosion in 21 (17.6%). Total device replacement was performed in 75 cases (47.2%). Of 119 patients undergoing secondary implantation 91 (76.5%) needed no additional surgical intervention, while 28 (23.5%) required a total of 40 surgical revisions for new mechanical (15 or 37.5%) and nonmechanical (25 or 62.5%) problems. Five-year durability outcomes for primary and secondary AUS implantation were comparable at 80% and 88%, respectively. Similarly excellent continence outcomes (0 to 1 pad daily) were noted in 90% and 82% of patients undergoing primary and secondary AUS implantation, respectively. Secondary and tertiary AUS revisions resulted in the restoration of baseline continence in 106 cases (89%). CONCLUSIONS: Our study suggests that outcomes for secondary AUS reimplantation are comparable to those of primary AUS implantation and salvage of a good outcome is always probable, even following multiple prior revisions and cuff erosion. PMID- 15758762 TI - Structural assessment of the urethral sphincter in women with urinary retention. AB - PURPOSE: The pathophysiology of urinary retention in women is generally unknown but a subgroup of women with urinary retention have been diagnosed as having so called primary disorder of sphincter relaxation on the basis of an abnormal urethral sphincter electromyogram. It was suggested this sphincter overactivity could lead to work hypertrophy of the urethral rhabdosphincter and in this study we looked for any evidence of such muscle fiber hypertrophy. MATERIALS AND METHODS: In 9 women 18 to 45 years old (mean age 31.6) with urinary retention and overactive urethral sphincter electromyogram, light and electron microscopy were used to examine core needle biopsies of the urethral rhabdosphincter taken under transvaginal ultrasound control. Of the 9 patients only 5 biopsies processed for light microscopy and 4 processed for electron microscopy contained striated urethral muscle fibers. The results of these biopsies were compared to the morphology of a control specimen from a postmenopausal woman without a history of urinary retention. RESULTS: On light microscopy the urethral rhabdosphincter fiber diameter did not differ among patients (mean average 7.6 mum), was less than that reported in the literature (15 to 20), but did not differ from that of the control (mean 9.9). In all patients electron microscopy showed excessive peripheral sarcoplasm with lipid and glycogen deposition, and sarcoplasmic accumulation of normal mitochondria. These ultrastructural abnormalities were not seen in the control. CONCLUSIONS: To our knowledge this is the first morphological description of the urethral rhabdosphincter in a subgroup of women with urinary retention. Mean rhabdosphincter fiber diameter was approximately the same in patients and controls. This study does not support the previous theory that urethral sphincter overactivity in a subgroup of women with urinary retention leads to work hyperplasia of urethral rhabdosphincter fibers. An alternative hypothesis is suggested. PMID- 15758763 TI - Pentosan polysulfate sodium therapy for men with chronic pelvic pain syndrome: a multicenter, randomized, placebo controlled study. AB - PURPOSE: We evaluated the efficacy and tolerability of pentosan polysulfate sodium (PPS) for the treatment of men with chronic pelvic pain syndrome (CPPS), National Institutes of Health (NIH) category III. MATERIALS AND METHODS: In a 16 week double-blind study 100 men with a clinical diagnosis of CPPS were randomized to receive 300 mg PPS or placebo 3 times daily. Clinical Global Improvement (CGI) was the primary outcome measure. Additional outcome measures were the NIH-Chronic Prostatitis Symptom Index (CPSI), Subjective Global Assessment and Symptom Severity Index assessment tools. RESULTS: Significantly more patients receiving PPS experienced moderate to marked improvement based on CGI assessment (18 or 37% vs 8 or 18%, p = 0.04). However, mean CGI scores were not significantly different between the PPS group (1.0) and placebo groups (1.0 vs 0.6, p = 0.107). All NIH CPSI domains suggested a positive effect for PPS and for total NIH-CPSI the difference approached statistical significance (-5.9 or 22% vs -3.2 or 12%, p = 0.068). The PPS group showed significantly greater improvement in NIH-CPSI quality of life domain scores than the placebo group (-2.0 or 22% vs -1.0 or 12%, p = 0.031). Of patients receiving PPS 67% and 80% of those receiving placebo completed the 16-week study. Diarrhea, nausea and headache were the most common adverse events. CONCLUSIONS: Pentosan polysulfate (900 mg daily) was more likely than placebo to provide relief for CPPS symptoms. PMID- 15758764 TI - Urologic diseases in America project: benign prostatic hyperplasia. AB - PURPOSE: Benign prostatic hyperplasia (BPH), the most common benign neoplasm in American men, is a chronic condition that is associated with progressive lower urinary tract symptoms and affects almost 3 of 4 men during the seventh decade of life. Approximately 6.5 million of the 27 million white men who are 50 to 79 years old in the United States in 2000 were estimated to meet the criteria for discussing treatment. MATERIALS AND METHODS: The analytical methods used to generate these results have been described previously. RESULTS: In 2000 approximately 4.5 million visits were made to physician offices to for a primary diagnosis of BPH and almost 8 million visits were made with a primary or secondary diagnosis of BPH. In the same year approximately 87,400 prostatectomies for BPH were performed in inpatients in nonfederal hospitals in the United States. While the number of outpatient visits for BPH increased consistently during the 1990s, there was a dramatic decrease in the use of transurethral prostatectomy, inpatient hospitalization and length of hospital stay for this condition. These trends reflect the changing face of medical management for BPH, ie increasing use of pharmacological agents and minimally invasive therapies. In 2000 the direct cost of BPH treatment was estimated to be $1.1 billion exclusive of outpatient pharmaceuticals. CONCLUSIONS: Given the impact that BPH has on quality of life and health care cost in millions of American men, additional research into risk factors, diagnostic and therapeutic resource use, and effectiveness and cost benefit of therapies are warranted. PMID- 15758765 TI - VTimed Voiding for the Management of Urinary Incontinence in Adults. PMID- 15758766 TI - An open-label pilot study of cannabis-based extracts for bladder dysfunction in advanced multiple sclerosis. PMID- 15758767 TI - Bladder training for urinary incontinence in adults. PMID- 15758769 TI - TVT and Sparc Suburethral Slings: A Case-Control Series. PMID- 15758768 TI - Deafferentation of the Urinary Bladder and Implantation of a Sacral Anterior Root Stimulator (SARS) for Treatment of the Neurogenic Bladder in Paraplegic Patients. PMID- 15758770 TI - Incidence of pubic osteomyelitis after bladder neck suspension using bone anchors. PMID- 15758771 TI - Chronic Inflammation in Benign Prostate Hyperplasia is Associated With Focal Upregulation of Cyclooxygenase-2, Bcl-2, and Cell Proliferation in the Glandular Epithelium. PMID- 15758772 TI - Modeling of the pH- and the Temperature-Dependent Deviations of the Free to Total PSA (Prostate Specific Antigen) Ratios for Clinical Predictability of Prostate Cancer and Benign Prostate Hyperplasia. PMID- 15758773 TI - The Effect of Combined Androgen Ablation on the Expression of alpha1A-Adrenergic Receptor in the Human Prostate. PMID- 15758774 TI - A new clinical method for the assessment of penile endothelial function using the flow mediated dilation with plethysmography technique. AB - PURPOSE: Penile endothelial function (EnF) is 1 of the major factors involved in the pathophysiology of erectile dysfunction. EnF assessment could offer crucial information on the etiology and degree of severity of cavernosal vascular pathology. In the present study we propose a new technique for the evaluation of penile EnF and test its applicability using strain gauge plethysmography. MATERIALS AND METHODS: A total of 23 healthy subjects (13 younger and 10 older than 40 years) with no history of erectile dysfunction were studied. The flow mediated dilation technique was applied to the arm and penis in both age groups for the assessment of EnF. Baseline blood flow and the sequential flow recordings after rapid cessation of 5 minutes of ischemia were obtained in both organs. RESULTS: Baseline flow in the penis was significantly higher (approximately 3 fold) than that in the forearm and was not affected by age in either organ. Both measures of penile EnF, ie area under the flow-time curve (AUC) and maximal flow obtained after ischemia were significantly lower in the older group compared to the younger group (p <0.01 and p <0.02, respectively). Individual penile AUC and maximal flow were significantly correlated with age (r = 0.55, p <0.01 and r = 0.50, p <0.02, respectively). Finally a positive, significant correlation existed between penile and forearm AUC (p <0.05, r = 0.48). CONCLUSIONS: The implementation of the flow mediated dilation technique using mercury strain gauge plethysmography is simple and applicable for the assessment of penile EnF. Endothelial function parameters in the penis were found to correlate with those in the forearm, thus support for the validity of the technique is given. Further strength for the validity of this procedure in the penis comes from the comparison between the forearm and penis, and the relation to subject age. PMID- 15758775 TI - Characterization of localized seminal vesicle amyloidosis causing hemospermia: an analysis using immunohistochemistry and magnetic resonance imaging. AB - PURPOSE: We evaluated the characteristics of seminal vesicle amyloidosis (SVA) associated with hemospermia by immunohistochemistry and magnetic resonance imaging (MRI) as well as the clinical course of hemospermia. MATERIALS AND METHODS: Of 56 patients with hemospermia 12 underwent transperineal biopsy of the seminal vesicle under transrectal ultrasound monitoring. SVA was proved in 4 men 48 to 59 years old by histological and immunohistochemical examinations of specimens obtained by biopsy. Two men presented with the first episode of hemospermia and 2 presented with recurrent hemospermia. MRI at 1.5 Tesla was performed while hemospermia persisted and after its resolution. Patients were followed for 10 to 86 months with regard to the duration of hemospermia, the time of its resolution and its recurrence. RESULTS: Amyloid deposits in the subepithelial tissue of the seminal vesicles were permanganate sensitive, and positive for lactoferrin and the amyloid P component but negative for amyloid A protein, lambda and kappa chains, and beta2-microglobulin. The seminal vesicles with obvious intravesicular hemorrhage on needle puncture were hyperintense on T1 weighted images. After hemospermia resolution T1-weighted images became diffusely hypointense. T2-weighted images were of low intensity, representing amyloid deposits. Hemospermia resolved spontaneously in all patients in an average of 14 months. Although disease recurred in 1 patient after 8 months of resolution, it disappeared after 11 months of recurrence. CONCLUSIONS: Localized SVA with hemospermia shows hypointensity on T2-weighted MRI. Hemospermia is spontaneously resolved with the transition from hyperintense to hypointense T1-weighted MRI. PMID- 15758776 TI - Management of hyperprolactinemia in patients receiving antipsychotics. PMID- 15758777 TI - Testosterone Treatment Improves Body Composition and Sexual Function in Men With COPD, in a 6-Month Randomized Controlled Trial. PMID- 15758778 TI - Selective facilitation of sexual solicitation in the female rat by a melanocortin receptor agonist. PMID- 15758779 TI - Sexual Function of LUTS Patients Before and After Neodymium Laser Prostatectomy and Transurethral Resection of Prostate. A Prospective, Randomized Trial. PMID- 15758780 TI - Comparison of the Pharmacological Effects of a Novel Selective Androgen Receptor Modulator (SARM), the 5alpha-Reductase Inhibitor Finasteride, and the Antiandrogen Hydroxyflutamide in Intact Rats: New Approach for Benign Prostate Hyperplasia (BPH). PMID- 15758781 TI - Comparison of the aneuploidy frequency in embryos derived from testicular sperm extraction in obstructive and non-obstructive azoospermic men. PMID- 15758782 TI - Can Biological or Clinical Parameters Predict Testicular Sperm Recovery in 47,XXY Klinefelter's Syndrome Patients? PMID- 15758783 TI - Urologic diseases in America project: trends in resource use for urinary tract infections in women. AB - PURPOSE: Urinary tract infection (UTI) is one of the most common clinical diagnoses in women. In this study we examined epidemiological, economic and health care use trends for UTI in women in the United States. MATERIALS AND METHODS: The analytical methods used to generate these results have been described previously. RESULTS: The lifetime risk for UTI in women is high (greater than 50%). Between 1988 and 1994 the overall lifetime prevalence of UTI was estimated to be 53,067/100,000 women. Prescribing patterns demonstrated an increase in the trend toward using fluoroquinolones as first line therapy for UTI, which was associated with increased costs. Composite data revealed that overall expenditures for the treatment of UTIs in women in the United States, excluding spending on outpatient prescriptions, were approximately 2.47 billion dollars in 2000. Diagnosis and treatment of UTI in women is performed in various clinical settings. Inpatient hospitalization for UTI care has generally decreased in younger women but increased in elderly women. There has been a sharp increase in emergency room use by younger women, which may reflect disparities in access to health insurance or primary care providers. Most outpatient care of women with UTIs is performed in physician offices. CONCLUSIONS: Analysis of health care use and economic data on UTIs in women revealed various interesting trends. These findings will help shape understanding of UTI treatment in relation to other urological disorders in women. The results raise various important future research questions. PMID- 15758784 TI - Urologic diseases in america project: trends in resource use for urinary tract infections in men. AB - PURPOSE: Various types of urinary tract infection (UTI) occur in men. In this study we examined health care use trends, including epidemiological and economic factors, for UTI in men in the United States. MATERIALS AND METHODS: The analytical methods used to generate these results have been described previously. RESULTS: Approximately 20% of all UTIs occur in men. Between 1988 and 1994 the overall lifetime prevalence of UTI was estimated to be 13,689/100,000 men. Orchitis rates, particularly in older men, are generally higher than those of cystitis or pyelonephritis. Approximately 10% of all inpatient care of men with UTI is for orchitis (12 to 14/100,000 population). Rates of outpatient hospital and physician office care for male UTI have increased in the last decade. Rates for emergency room care for UTI in elderly men (85 to 94 years old) were almost twice those in men younger than 85 years. The adjusted mean health care expenditure for privately insured men with UTI was 5,544 dollars in 1999 compared to 2,715 dollars for men without UTI. Total annual health care expenditures for men and women with UTI were 5,544 dollars and 5,407 dollars, respectively. Mean time lost from work was slightly higher for men. Based on composite data overall medical expenditures for men with UTI in the United States were estimated to be approximately 1.028 billion dollars in 2000. CONCLUSIONS: Health care use and economic data on UTIs in men revealed a number of intriguing trends. These results raise various important questions for future research. PMID- 15758785 TI - Urologic diseases in America project: urinary incontinence in women-national trends in hospitalizations, office visits, treatment and economic impact. AB - PURPOSE: We describe temporal trends in hospitalizations, outpatient visits and the treatment of female urinary incontinence (UI), and estimated the costs of incontinence using national databases. MATERIALS AND METHODS: The analytic methods used to generate these results have been described previously. RESULTS: The rate of hospitalization with a primary diagnosis of UI decreased from 51/100,000 women in 1994 to 44/100,000 in 2000 and mean length of stay decreased from 3.1 days to 2.1. In contrast, outpatient visits for UI more than doubled during the same period from 845/100,000 women to 1,845/100,000. Rates of inpatient surgical treatment for UI decreased slightly from 1994 to 2000, while ambulatory surgical center visit rates for Medicare beneficiaries 65 years or older more than doubled from 60/100,000 in 1992 to 142/100,000 in 1998. Medical expenditures for UI increased substantially during the 1990s, almost doubling from 128.1 million dollars in 1992 to 234.4 million dollars in 1998 for Medicare beneficiaries 65 years or older. This increase was due almost entirely to increased outpatient costs, which increased from 25.4 million dollars or 9.1% of total costs in 1992 to 329 million dollars or 27.3% of total costs in 2000 in this group. CONCLUSIONS: While existing national databases generally capture only the minority of incontinent women with UI who seek and receive care for UI, they are useful for documenting treads in service use and surgical treatments, and estimating economic impact. This data can be helpful when formulating public policy and designing observational and clinical studies. PMID- 15758786 TI - Urologic diseases in America project: urinary incontinence in males--demographics and economic burden. AB - PURPOSE: We quantified and describe the demographics and economic burden of male urinary incontinence in the United States of America. MATERIALS AND METHODS: The analytic methods used to generate these results have been described previously. RESULTS: Urinary incontinence (UI) affects men of all ages, including 17% of males older than 60 years in the United States, which is an estimated 3.4 million men. There is a strong trend toward an increasing prevalence of UI with increasing age as well as an increase in the prevalence of UI in males with time. Ethnicity has less of a role in prevalence estimates in men than in women. The largest impact of UI in elderly men is in physician office visits, followed by outpatient services and surgeries. Resource use is greatest in the nursing home setting, where more than half of men have UI and require assistance with toileting. The overall economic burden for male UI is estimated at 18.8 billion dollars in direct medical costs in 1998/1999 dollars. Medical expenditures for UI for male Medicare beneficiaries 65 years and older have doubled since 1992. Compared to persons without UI the presence of UI increases the annual expenditures per person yearly from 3,204 dollars to 7,702 dollars. CONCLUSIONS: The direct and indirect costs of male UI increased throughout the 1990s with annual expenditures per person yearly in men with UI more than double that in men without UI. Given the aging population and staggering impact of UI in nursing home settings, there is a compelling need for further research into effective prevention, treatment and management strategies. PMID- 15758787 TI - Economic costs of benign prostatic hyperplasia in the private sector. AB - PURPOSE: Several studies document the impact of benign prostatic hyperplasia (BPH) in working, aged men. Direct medical costs related to BPH treatment are largely borne by employees through higher premiums. However, indirect costs related to lost work are primarily borne by the employer. In this study we used claims data and absentee records from large employers to estimate the costs associated with BPH in working age males. MATERIALS AND METHODS: We used 2 data sources to examine direct and indirect costs associated with BPH in a privately insured, nonelderly population. Multivariate regression models were used to predict spending for persons with and without a medical claim for BPH, controlling for relevant covariates. Data on work loss were linked to medical claims to estimate work loss related to treatment for BPH. RESULTS: Mean annual expenditures were 4,193 dollars for men without a medical claim for BPH. In contrast, annual spending was 5,729 dollars for men with a claim for BPH. Thus, the incremental cost associated with a diagnosis of BPH was 1,536 dollars yearly. Overall the average employee with the condition missed 7.3 hours of work yearly related to BPH with approximately 10% reporting some work loss related to a health care encounter for BPH. CONCLUSIONS: Treatment of men with BPH places a significant burden on employees and their employers through direct medical costs as well as through lost work time. Direct and indirect costs to the private sector related to BPH treatment are estimated to be 3.9 billion dollars. PMID- 15758788 TI - Clinical outcomes after sexuality preserving cystectomy and neobladder (prostate sparing cystectomy) in 44 patients. AB - PURPOSE: We describe the functional outcome on erectile function, continence and voiding, and local and distant cancer recurrence rates in 44 patients after sexuality preserving cystectomy and neobladder (prostate sparing cystectomy). MATERIALS AND METHODS: A total of 44 males underwent cystectomy with preservation of the prostate, seminal vesicles and vasa deferentia, after which a Studer type neobladder was anastomosed to the prostate. Oncological outcome (disease specific survival, distant and local recurrence rates) and functional results (continence, voiding, erectile function) were determined. RESULTS: At a median followup of 42 months, 13 (30%) patients died of cancer. All 13 experienced widespread disease, which was combined with a pelvic recurrence (pelvic recurrence rate 6.9%) in 3. The 3-year survival according to pathological stage was 86% for pT 2N0 or lower, 63% pT3N0 and 39% for node positive tumors (anyT Npos). Prostate cancer was diagnosed in 1 patient 5 years after treatment, and recurrent carcinoma in situ in the prostatic urethra in another patient. Complete daytime and nighttime continence was achieved in 95.3% and 74.4%, respectively. Incontinence during day and night could be managed by 1 pad per day/night in 4.7% and 20.9%, respectively, while 4.7% needed more than 1 pad per night. Erectile function could be determined in 40 patients, and potency was maintained in 77.5%, impaired in 12.5% and absent in 10%. CONCLUSIONS: Functional results with regard to erectile function and urinary continence after prostate sparing cystectomy are good. Oncological results have been promising, but need to be confirmed after longer followup and in larger trials. PMID- 15758789 TI - Health related quality of life after radical cystectomy and urinary diversion for bladder cancer: a systematic review and critical analysis of the literature. AB - PURPOSE: Continent forms of urinary diversion have become a gold standard of urinary tract reconstruction after radical cystectomy, based mostly on the premise of improved quality of life. It is unclear whether the existing body of literature supports this assumption. MATERIALS AND METHODS: We performed a systematic review to determine if differences exist in health related quality of life (HRQOL) outcomes among different types of urinary diversion after radical cystectomy. A MEDLINE search was performed inclusive of the dates 1966 to January 2004. Inclusion criteria included adult patients, patients with bladder cancer, comparative studies, original research, primary study outcome related to quality of life, and use of a quality of life instrument to measure outcomes. Only studies comparing neobladder, continent reservoir, or conduit diversion were included. RESULTS: Of 378 initial articles 15 studies met all inclusion criteria. None of the studies were randomized trials. Only 1 study was prospective. Of 15 studies 10 (67%) used some type of previously validated HRQOL instrument, 10 (67%) used some form of ad hoc instrument, 11 (73%) used bladder cancer disease specific instruments, while 9 (60%) used general instruments. Only 1 of the bladder cancer specific instruments had been previously validated. Common limitations included unvalidated HRQOL instruments, use of general HRQOL instruments only, lack of baseline data, cross- sectional analysis, and retrospective study design. CONCLUSIONS: To date, the current body of published literature is insufficient to conclude that 1 form of urinary diversion is superior to another based on HRQOL outcomes. Future studies should attempt to incorporate prospective data collection, longer term followup and validated disease specific HRQOL instruments. PMID- 15758790 TI - Mental health of low income uninsured men with prostate cancer. AB - PURPOSE: We evaluated mental health outcomes in a cohort of low income, uninsured men with prostate cancer and identified factors that influence mental health. MATERIALS AND METHODS: We performed a retrospective cohort study of 277 subjects enrolled in a program that provides free care to men with prostate cancer who have an annual income of no more than 200% of the federal poverty level. We compared scores on the 5-item RAND Mental Health Inventory (MHI-5) to those in individuals with other chronic diseases. We also examined the relationship between MHI-5 scores and validated measures of general and disease specific health related quality of life. Disease specific quality of life included measures of distress related to urinary, sexual and bowel habits. Multivariate analyses were performed to evaluate factors associated with mental health score. RESULTS: Most men studied were Hispanic (51.6%) and had at most a high school education (85.9%). Mean MHI-5 score +/- SD was 68 +/- 23 on a 100-point scale, significantly worse than cohorts of men with diabetes, congestive heart failure and chronic pulmonary disease. Hispanic ethnicity, urinary bother and bowel bother were negatively associated with mental health. Spirituality and physical functioning were positively associated with mental health. CONCLUSIONS: Economically disadvantaged men with prostate cancer report worse mental health than people with other chronic diseases. Patients especially at risk are those with significant urinary or bowel distress, poor physical health, low spirituality and Hispanic ethnicity. PMID- 15758791 TI - Variation in Urological Practice Amongst Spinal Injuries Units in the UK and Eire. PMID- 15758792 TI - The economic impact of chronic prostatitis. PMID- 15758793 TI - Development and Validation of the Self-Esteem and Relationship (SEAR) Questionnaire in Erectile Dysfunction. PMID- 15758794 TI - Traumatic adrenal injuries. AB - PURPOSE: We determined the prevalence, management and general prognosis of blunt isolated and nonisolated adrenal injuries in a pediatric population. MATERIAL AND METHODS: We analyzed trauma data from a pediatric institution for the period 1991 to 1998 to identify patients with blunt traumatic adrenal injuries diagnosed by computerized tomography. We then performed a detailed chart review to obtain data on the presence of concomitant intra-abdominal and extra-abdominal injuries, transfusion requirements, intensive care unit monitoring requirements, hospital course and the presence of followup studies. RESULTS: Among 9,199 pediatric trauma cases we identified 20 adrenal injuries (0.22%), of which 15 (75%) were nonisolated and 5 (25%) were isolated. The right adrenal gland was injured in 17 (85%) of the 20 patients. In the 15 nonisolated adrenal injuries concomitant injury to the liver (13 cases, 87%) and ipsilateral kidney (8 cases, 53%) were most common. Three (60%) of the 5 patients with isolated adrenal injury required transfusion for adrenal hemorrhage. No patient required intensive care unit monitoring or operative intervention. Only 2 (10%) of the 20 patients underwent followup computerized tomography, both of whom had resolution of the adrenal injury. CONCLUSIONS: In the pediatric population blunt adrenal injuries are rare and typically present as part of a multiorgan trauma. The right adrenal gland is more likely to be injured, with liver trauma as the most commonly associated injury, followed by ipsilateral renal injury. Although there is a possibility of significant adrenal hemorrhage requiring transfusion, adrenal trauma is typically self-limited and does not require intensive care monitoring or operative intervention. PMID- 15758795 TI - A novel approach to skin coverage for epispadias repair. AB - PURPOSE: Current techniques for epispadias repair have resulted in significant improvement in the reconstruction of the urethra, corpora and glans. The final challenging step is to enhance the cosmetic result by accomplishing skin coverage for the penis and subpubic area, creating a penopubic and penoscrotal angle without dorsal suture lines, and avoiding future dorsal tethering of the penis as a result of scar contraction. We report a novel technique for penile skin coverage in the patient with epispadias that results in a superior cosmetic outcome. MATERIALS AND METHODS: The technique involves creation of 2 flaps-a ventral preputial transverse island flap rotated dorsally to cover the dorsal aspect of the penile shaft, and an advancement flap from the patch of skin present between the penis and scrotum in epispadias, which is advanced distally to cover the ventral aspect of the penis. The 2 flaps are sewn to each other with 2 lateral suture lines. Thus, the dorsal and ventral aspects of the penis are covered with intact skin devoid of suture lines. RESULTS: This technique was used in 8 males 2 days to 15 years old. The epispadias was part of exstrophy in 5 patients and an isolated defect in 3. Both flaps healed well in 7 of 8 patients. In 1 exstrophy case a segment of the transverse island flap became ischemic and was discarded intraoperatively. Dorsal skin coverage in this patient was achieved using a laterally based flap from the inguinal area, which healed without problem. Followup was 6 to 33 months. No patient had development of skin tethering, curvature or recurrence of the dorsal chordee. The cosmetic appearance of the penis was subjectively superior to that of boys who underwent skin closure using reverse Byars flaps. CONCLUSIONS: The cosmetic appearance of the penis using this novel technique is superior because of the absence of the dorsal scar that may cause chordee, the development of penopubic and penoscrotal angles, which gives the penis a more normal appearance, and the absence of the redundant patch of skin between the shaft of the penis and the scrotum. PMID- 15758796 TI - Tolterodine treatment for children with symptoms of urinary urge incontinence suggestive of detrusor overactivity: results from 2 randomized, placebo controlled trials. AB - PURPOSE: We report the results of the first 2 large randomized controlled trials designed to evaluate the efficacy and safety of tolterodine extended release in children 5 to 10 years old with symptoms of urinary urge incontinence suggestive of detrusor overactivity. MATERIALS AND METHODS: Two double-blind, placebo controlled trials were conducted sequentially. Children 5 to 10 years old with incontinence suggestive of detrusor overactivity (1 or more diurnal incontinence episodes per 24 hours) were randomized to tolterodine (2 mg daily) or placebo for 12 weeks. The primary end point was the change from baseline to week 12 in the number of incontinence episodes per week. Changes from baseline in the number of voids per 24 hours and volume of urine per void were also evaluated. Exploratory analyses were conducted to determine whether particular subsets of patients showed differential responses to treatment. RESULTS: A total of 224 and 487 children (mean age 8 years) were randomized to placebo and tolterodine, respectively. Differences in the number of incontinence episodes per week, voids per 24 hours, and volume of urine per void between tolterodine and placebo did not reach statistical significance. This finding may be explained by a high placebo response and under dosage of tolterodine among children with greater body weight. Tolterodine was well tolerated. CONCLUSIONS: Analysis of the primary efficacy outcome did not reveal a statistically significant effect of treatment. However, secondary analyses demonstrated that tolterodine was well tolerated among 5 to 10-year-old children with diurnal incontinence. Exploratory analyses also showed that children weighing 35 kg or less with detrusor overactivity characterized by incontinence and/or frequent voiding benefited most from tolterodine treatment, suggesting that a weight adjusted dosing regimen may be required for optimal response among older and heavier children. PMID- 15758797 TI - The Macedo-Malone antegrade continence enema procedure: early experience. AB - PURPOSE: The successful treatment of fecal incontinence can dramatically improve the quality of life of affected children. The introduction of the Malone antegrade continence enema provides the opportunity to manage previously resistant cases. However, using the to create this catheterizable channel is not always possible, and the duration of these antegrade enemas is a source of concern for the patients. We describe a new approach to create left continent colonic access to shorten the duration of these enemas, and report the experience gained from the first 9 cases managed at our institution. MATERIALS AND METHODS: During a 5-year period 9 patients underwent a Macedo-Malone antegrade continence enema at our institution. Incontinence was associated with myelomeningocele in 7 patients and anorectal malformation in 2. The antegrade continence enema procedure is begun by isolating a 2 cm flap in a tenia on the left colon (spleen flexure). A 12Fr silicone Foley catheter is placed on the mucosal surface of the flap to allow tubularization of the plate with interrupted polyglycolic acid 3 zero transverse sutures, creating an efferent tubular conduit. Antegrade colonic washouts were started 2 weeks after surgery with saline solution or tap water in all patients. RESULTS: Followup of our 9 cases ranged from 8 to 33 months (average 20.7). Enema volume varied from 250 to 800 ml, with administration taking from 45 to 60 minutes, and colonic evacuation occurred within 30 to 60 minutes of enema administration. Of the 9 patients 8 were completely continent and 1 was partially continent. Four patients experienced difficulty with catheterization initially because of stenosis of the stomal track. The affected stomas were dilated, which was successful in 1 case. Three patients subsequently required stomal revision. CONCLUSIONS: The Macedo-Malone procedure is a relatively straightforward operative approach providing an effective washout technique that is acceptable to parents and children. PMID- 15758798 TI - Post-pyelonephritic renal scars are not associated with vesicoureteral reflux in children. AB - PURPOSE: Children with pyelonephritis are at risk for renal damage. We assess the value of clinical signs and urological abnormalities in predicting renal scars in children following pyelonephritis. MATERIALS AND METHODS: A total of 64 hospitalized children (29 females and 35 males, median age 2.9 years) underwent ultrasonography and technetium labeled dimercapto-succinic acid (DMSA) scintigraphy imaging within 1 week following the diagnosis of the first pyelonephritis. Voiding cystourethrography was performed 8 weeks after the diagnosis. Followup DMSA scintigraphy was performed in 58 patients after 2 years of followup. RESULTS: Urological abnormalities observed were vesicoureteral reflux (VUR, grade 2 or higher) in 11 patients (19%), nonrefluxing and nonobstructed megaureter in 2 (4%) and pyeloureteral obstruction in 1 (2%). The first DMSA scintigraphy showed parenchymal defects in 48% of patients. VUR did not increase the risk of renal defects. At 2 years after the infection 12 of the 58 patients (21%) had renal scars. Nine of these patients did not have VUR. However, 2 patients with high grade VUR and repeat infections demonstrated deterioration of kidney function during followup. The patients with renal scars were older than those without scars (3.1 vs 0.8 years, p = 0.0291) at the time of infection. CONCLUSIONS: Renal scars after first pyelonephritis are in most cases not associated with abnormalities of the urinary tract, but are caused by the infection itself. However, structural abnormalities may predispose to recurrent infections. Following pyelonephritis new renal scars may develop in all age groups in both sexes. PMID- 15758799 TI - Does prenatal diagnosis influence the morbidity associated with left in situ nonfunctioning or poorly functioning renal moiety after endoscopic puncture of ureterocele? AB - PURPOSE: We compared long-term morbidity associated with left in situ nonfunctioning or poorly functioning renal moiety of a duplex system in children with prenatal vs postnatal diagnosis of ureterocele who underwent endoscopic puncture. MATERIALS AND METHODS: A total of 48 children underwent primary endoscopic puncture of duplex system ureterocele. Of the cases 35 (73%) were diagnosed prenatally (group 1) and 13 (27%) postnatally (group 2). Median age at time of puncture was 4 months in group 1 and 3.5 years in group 2. A total of 20 patients in group 1 (57%) and 8 in group 2 (62%) presented with intravesical ureterocele, while 15 in group 1 (43%) and 5 in group 2 (38%) had ectopic ureterocele. A total of 20 children in group 1 (57%) and 7 in group 2 (54%) had a nonfunctioning renal moiety, and 15 in group 1 (43%) and 6 in group 2 (46%) had a poorly functioning ureterocele moiety. Vesicoureteral reflux (VUR) was present in 23 children in group 1 (66%) comprising 30 renal refluxing units (RRUs), and in 12 in group 2 (92%) comprising 14 RRUs. Median followup was 9 years (range 1 to 15) for both groups. RESULTS: Preoperative urinary tract infection (UTI) was common in group 2 (92%) vs group 1 (20%). No patient in group 1 had development of UTI after puncture, while 23% of the children in group 2 presented with UTI. Four children (2 from each group) with ectopic ureterocele required secondary puncture resulting in satisfactory drainage. A total of 14 RRUs (47%) showed spontaneous resolution of VUR in group 1 compared to 3 (21%) in group 2. Four RRUs (13%) required endoscopic correction due to high grade VUR in group 1. Two RRUs (17%) were treated with endoscopic correction and 2 (17%) with ureteral reimplantation due to UTI in group 2. Only 1 patient in group 1 underwent nephrectomy due to nonfunctioning kidney, while 2 patients in group 2 required partial nephrectomy due to UTI. CONCLUSIONS: Our data reveal that prenatal diagnosis of duplex system ureterocele is associated with fewer UTIs, and early endoscopic management may decrease UTI and the need for additional surgery. Nonfunctioning or poorly functioning renal moieties left in situ following successful endoscopic decompression of ureterocele are not associated with additional morbidity and do not require partial nephrectomy in the majority of the cases. PMID- 15758800 TI - Prenatally detected primary megaureter: a role for extended followup. AB - PURPOSE: A high rate of spontaneous resolution of or decrease in urinary tract dilatation is expected for most cases of primary megaureter (PM). We analyzed our results with extended long-term nonsurgical treatment in a group of children diagnosed with prenatally detected PM. We also sought to determine the long-term safety of this approach and the clinical significance of residual hydroureteronephrosis (HUN) when complete resolution does not occur. MATERIALS AND METHODS: We reviewed our experience with 40 infants with prenatally detected hydronephrosis diagnosed as PM who were initially treated nonoperatively between 1986 and 1999. Patients were followed with ultrasonography, nuclear renography and, in some instances, excretory urogram until stable improvement or complete resolution of HUN was noted. Records for patients who specifically returned for long-term followup history and radiological imaging were also reviewed. RESULTS: Of 40 patients with 57 megaureters (17 with bilateral PMs) who were followed 4 infants (10%) with severe HUN were diagnosed with diminished renal function by nuclear renography and underwent surgical repair. A total of 27 infants (67.5%, 40 megaureters) with PM who were followed without surgical intervention returned for followup examination and imaging for a minimum of 24 months after initial diagnosis, for a mean of 6.8 years (range 24 to 210 months). Ultrasonography in these patients revealed complete resolution (Society for Fetal Urology grade 1 or less hydronephrosis) in 21 megaureters (52.5%) at a mean of 2.9 years and improved or stable HUN in 19 megaureters (47.5%). Antibiotic prophylaxis was administered to 12 patients beyond the first month of life and discontinued at a mean age of 19.9 months. A subgroup of 10 patients (16 megaureters) from this cohort returned for extended followup imaging at a mean of 13.4 years (range 84 to 210 months) following initial diagnosis. Four patients with documented resolution of HUN had no recurrence during long-term followup. One male with bilateral PMs diagnosed at birth that had been stable but were not resolved at 8 year followup suffered significant worsening in the degree of HUN, diminished renal function on 1 side and a renal stone on the contralateral renal unit at 14 year followup, despite remaining asymptomatic. CONCLUSIONS: Long-term followup of children with prenatally diagnosed PM with mild to moderate hydronephrosis confirms a high incidence of resolution and improvement. As we have documented 1 case of worsening HUN at 14-year followup, we believe that ultrasonography should be periodically continued until the child reaches adulthood, at least in instances where complete resolution of HUN is not documented. PMID- 15758801 TI - Refluxing ureteral reimplant as temporary treatment of obstructive megaureter in neonate and infant. AB - PURPOSE: An obstructive megaureter identified in the neonatal period can be managed using a number of techniques, with the primary goal being to minimize the potential for further injury to the affected kidney. We describe our experience with refluxing ureteral reimplantation as a novel method for temporizing the obstructive megaureter. MATERIALS AND METHODS: Three patients identified prenatally with severe hydroureteronephrosis were confirmed following delivery to have an obstructive ectopic ureter. Unilateral obstruction was identified in 2 patients (1 female, 1 male). The third patient was a female with bilateral single system ectopic ureters. Treatment consisted of anastomosing the ureter proximal to the obstruction to the dome of the bladder in a freely high grade refluxing fashion. All of the patients were placed on antibiotic suppression after surgery. RESULTS: All patients demonstrated improved drainage of the affected kidney(s) following surgery. One female patient with unilateral obstruction had a poorly functioning kidney that showed no improvement of renal function 6 months following refluxing reimplantation, and laparoscopic nephrectomy was performed. The male patient with unilateral obstruction had adequate function with a significantly decreased ureteral diameter 1 year following refluxing ureteral reimplant, and a ureteral reimplantation without tapering was performed. The female patient with bilateral obstruction had 1 breakthrough urinary tract infection 6 months after surgery and now awaits second stage repair. CONCLUSIONS: Refluxing ureteral reimplantation is a safe, easy, beneficial and well tolerated means of temporizing the obstructive megaureter. This technique allows time for the child to mature, while accurately establishing renal function and preparing for a definitive surgical solution. PMID- 15758802 TI - Highly potent and moderately potent topical steroids are effective in treating phimosis: a prospective randomized study. AB - PURPOSE: We report a prospective randomized study comparing the effects of highly potent and moderately potent topical steroids in treating pediatric phimosis. MATERIALS AND METHODS: A total of 70 boys 1 to 12 years old with phimosis were randomly assigned to receive topical application of either betamethasone valerate 0.06% (a highly potent steroid) or clobetasone butyrate 0.05% (a moderately potent steroid). Parents of the boys were instructed to retract the foreskin gently without causing pain, and to apply the topical steroids over the stenotic opening of the prepuce twice daily for 4 weeks, then for another 4 weeks if no improvement was achieved. Retractibility of the prepuce was graded from 0 to 5. Response to treatment was arbitrarily defined as improvement in the retractibility score of more than 2 points. RESULTS: Mean treatment and followup periods were 4.3 and 19.1 weeks, respectively. The response rates in boys treated with betamethasone valerate and clobetasone butyrate were 81.3% and 77.4%, respectively (p = 0.63). Mean retractibility score decreased from 3.9 +/- 1.0 to 1.7 +/- 1.1, and 4.2 +/- 1.0 to 1.9 +/- 1.0 in the betamethasone and clobetasone groups, respectively. Both steroids were effective in all age groups. Pretreatment retractibility score did not affect treatment outcomes. No adverse effect was encountered. CONCLUSIONS: Highly potent and moderately potent topical steroids are of comparable effectiveness in treating phimosis. A less potent steroid may be considered first to decrease the risk of the potential adverse effects. PMID- 15758803 TI - DMSA Study Performed During Febrile Urinary Tract Infection: A Predictor of Patient Outcome? PMID- 15758804 TI - Ovarian torsion: to pex or not to pex? Case report and review of the literature. PMID- 15758805 TI - Rhabdomyosarcoma of the bladder and prostate. PMID- 15758806 TI - Arterial pseudoaneurysm after extracorporeal shock wave lithotripsy. PMID- 15758807 TI - The effects of intentional cryoablation and radio frequency ablation of renal tissue involving the collecting system in a porcine model. AB - PURPOSE: Ablative techniques for the treatment of urological malignancy are gaining acceptance and they are likely to become more widely used in clinical practice. Indications and limitations of the technologies are still evolving. In a porcine model we evaluated the safety and efficacy of cryotherapy and radio frequency ablation (RFA) of cortical and deep renal tissue. MATERIALS AND METHODS: In 11 swine argon gas based cryoablation or RFA of renal tissue adjacent to the collecting system was performed using a laparoscopic or percutaneous approach. Lesions created in renal units 30 days or 2 hours prior to harvest were termed chronic or acute. Using single or multiple 17 gauge cryoneedles or 3.0 mm cryoprobes and 2 freeze-thaw cycles (10-minute freeze and 5-minute thaw) 13 acute and 10 chronic cryolesions were made. Using a single 16 gauge umbrella-shaped RFA probe and 2 heating cycles to maximum impedance 13 acute and 4 chronic RFA lesions were made. Gross and microscopic tissue analysis was performed to assess lesion size and renal parenchymal, collecting system and arterial effects. Acute cryolesion size estimation by laparoscopic or transcutaneous ultrasound (US) was compared with pathological lesion size. RESULTS: Acute cryolesions on hematoxylin and eosin staining demonstrated uniform coagulative necrosis of renal parenchyma and chronic cryolesions demonstrated uniform necrosis with fibrous scar formation. Interlobar artery (adjacent to renal pyramid) preservation occurred in 7 of 13 acute and 5 of 9 chronic cryolesions. Urothelial architecture was preserved in 8 of 13 acute and 7 of 9 chronic cryolesions. Acute and chronic RFA lesions demonstrated indeterminate necrosis on hematoxylin and eosin staining, although triphenyl tetrazolium chloride staining of gross specimens confirmed necrosis most definitively in renal cortex. Interlobar artery preservation occurred in 6 of 13 acute and 3 of 4 chronic RFA lesions. Urothelial architecture was preserved in 1 of 13 acute and 2 of 4 chronic RFA lesions. Acute cryolesion dimensions measured by laparoscopic US equaled or underestimated lesion size measured grossly in all 6 cases. Lesion dimensions measured by transcutaneous US equaled or underestimated true lesion size in 3 of 6 cases. In 3 of 6 lesions transcutaneous US overestimated true lesion size by 20%, 76% and 260%, respectively. CONCLUSIONS: Renal cortical tissue can be effectively destroyed by cryoablation or RFA. However, treatment of deep parenchymal lesions with either modality may result in incomplete ablation. Cryosurgery but not RFA spares the collecting system in an acute setting. However, healing or regrowth of the urothelium may occur with time after RFA. Laparoscopic US is more accurate for cryolesion monitoring than transcutaneous US. PMID- 15758809 TI - Interleukin-8 response in cells from the human urinary tract induced by lipopolysaccharides of Proteus mirabilis O3 and O18. AB - PURPOSE: Proteus mirabilis is a common pathogen associated mainly with complicated urinary tract infections and sometimes with septicemia. There is great serological diversity of the microorganism. While P. mirabilis O3 is commonly found in patients with infections, the serotype O18 rarely occurs. The O18 lipopolysaccharide contains a phosphocholine substitute, which makes it unique among Proteus strains. To explain different clinical significance of the strains we evaluated the biological activity of the lipopolysaccharides of P. mirabilis O3 and O18, as measured by interleukin-8 (IL-8) production. MATERIALS AND METHODS: Three cell lines were used, namely uroepithelial cells, renal epithelial cells and monocytes. IL-8 production was determined on the protein and mRNA levels using enzyme-linked immunosorbent assay and real-time polymerase chain reaction, respectively, and CD14 expression on the cell surface was studied using flow cytometry. RESULTS: Uroepithelial cells and monocytes reacted to lipopolysaccharides by higher IL-8 production compared with renal epithelial cells. Cell specific IL-8 response corresponded to the cell expression of CD14. Renal epithelial cells produced more IL-8 after stimulation with the phosphocholine rich lipopolysaccharide O18, although adding phosphocholine alone did not induce or increase IL-8 production. CONCLUSIONS: Our data suggest that different cells within the human urinary tract have different roles during urinary tract infection. The biological activity and pathogenicity of P. mirabilis lipopolysaccharides might be determined by their specific chemical structure. PMID- 15758808 TI - Effect of hyperthermia on the cytotoxicity of 4 chemotherapeutic agents currently used for the treatment of transitional cell carcinoma of the bladder: an in vitro study. AB - PURPOSE: Hyperthermia combined with chemotherapy is not a novel cancer treatment. However, the working mechanism of this combination therapy is not fully understood. In the current in vitro study we investigated the differences in cytotoxicity of 4 chemotherapeutic agents at 37C or 43C. MATERIALS AND METHODS: The human transitional cell carcinoma cell lines used were RT4, RT112, 253J and T24. Cells were seeded in 96-well microtiter plates. After 24 hours cells were treated for 60 minutes with increasing concentrations of mitomycin C, epirubicin, gemcitabine and EO9 at a temperature of 37C or 43C. After treatment cells were rinsed 3 times and left for 24 hours in the incubator at 37C. The influence of chemotherapy and temperature on cell survival was determined by MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) assay. RESULTS: Decreased cell proliferation with increasing concentrations of chemotherapeutic agents was demonstrated. EO9 proved to be the most potent agent at each temperature. Hyperthermia alone did not demonstrate decreased cell proliferation. However, a synergistic effect on decreased cell proliferation was demonstrated in all cell lines and chemotherapeutic agents used, although each had a maximum at a different chemotherapy concentration and to a different extent. Synergism was most obvious in cell lines treated with low dose epirubicin. CONCLUSIONS: Synergism with hyperthermia and chemotherapy was clearly demonstrated for epirubicin, EO9, mitomycin C and to a lesser extent gemcitabine. Hyperthermia alone did not cause decreased cell proliferation. Synergism was most prominent with low drug doses and the most potent drug used in this in vitro study was EO9. PMID- 15758810 TI - Morphology and localization of interstitial cells in the guinea pig bladder: structural relationships with smooth muscle and neurons. AB - PURPOSE: In the current study we examined the location of interstitial cell of Cajal (ICC)-like cells in the guinea pig bladder wall and studied their structural interactions with nerves and smooth muscle cells. MATERIALS AND METHODS: Whole mount samples and cryosections of bladder tissue were labeled with primary and fluorescent secondary antibodies, and imaged using confocal and multiphoton microscopy. RESULTS: Kit positive ICC-like cells were located below the urothelium, in the lamina propria region and throughout the detrusor. In the suburothelium they had a stellate morphology and appeared to network. They made connections with nerves, as shown by double labeling experiments with anti-kit and anti-protein gene product 9.5. A network of vimentin positive cells was also found, of which many but not all were kit positive. In the detrusor kit positive cells were most often seen at the edge of smooth muscle bundles. They were elongated with lateral branches, running in parallel with the bundles and closely associated with intramural nerves. Another population of kit positive cells was seen in the detrusor between muscle bundles. These cells had a more stellate-like morphology and made connections with each other. Kit positive cells were seen tracking nerve bundles and close to intramural ganglia. Vimentin positive cells were present in the detrusor, of which some were also kit positive. CONCLUSIONS: There are several populations of ICC-like cells throughout the guinea pig bladder wall. They differ in morphology and orientation but all make connections with intramural nerves and in the detrusor they are closely associated with smooth muscle cells. PMID- 15758811 TI - Shock wave lithotripsy of stones implanted in the proximal ureter of the pig. AB - PURPOSE: Ureteral stones can be difficult to treat with shock wave (SW) lithotripsy. A strategy for lithotripsy of proximal ureteral stones is to push them back into the renal pelvis prior to administering SWs. However, push-back is invasive and not always possible. Since there are few clues to suggest how best to treat ureteral stones with SWs in situ, we developed an animal model for research on lithotripsy for ureteral stones. MATERIALS AND METHODS: Gypsum model stones were implanted bilaterally in the proximal ureter and renal calix of the pig via percutaneous access. Lithotripsy was performed using a HM3 lithotripter (Dornier Medical Systems, Marietta, Georgia) and stones at each location were treated with the same dose (400 SWs, 20 kV and 30 SWs per minute). Fragments were collected and the percent increase in projected surface area of the particles was determined. RESULTS: The breakage (mean percent area increase) of stones implanted in the proximal ureter was significantly less than that of stones located in the renal calix treated with the same dose of shock waves (134% vs 327%, p <0.001). Also, stones that were fully confined by the ureter did not break as well as stones located at the ureteropelvic junction. This indicates that the physical environment surrounding a stone can have a significant effect on the efficiency of SW action. CONCLUSIONS: The observation that stones implanted in the ureter showed decreased breakage compared with stones in the kidney is consistent with clinical experience. This finding is a valuable and even essential prerequisite for any experimental animal model system intended for the study of SW action in the breakage of ureteral stones. PMID- 15758813 TI - Effect of age on rabbit bladder function and structure following partial outlet obstruction. AB - PURPOSE: We determined whether young and old rabbits respond differently to partial bladder outlet obstruction. MATERIALS AND METHODS: A total of 16 male New Zealand White rabbits were separated into 2 groups of 8 each. Group 1 consisted of young rabbits (age 7 weeks) and group 2 consisted of old rabbits (age 2 years). Four rabbits per group underwent partial outlet obstructions and 4 underwent sham operation. Four weeks following surgery individual bladder strips were used for contractile studies and the remaining tissue was examined histologically. RESULTS: Contractile responses to all forms of stimulation between the young and old sham operated groups were similar. Contractile responses to all forms of stimulation were significantly decreased to the same degree in the 2 obstructed groups. However, the rate of tension generation to field stimulation was decreased to a significantly greater degree in young vs old bladders. Although young and old bladders showed smooth muscle hypertrophy, older rabbits showed significantly greater thickening of the serosa than young rabbits. Young rabbits showed significant inflammation, hemorrhage and expansion of the lamina propria, whereas old rabbits showed none of these characteristics. CONCLUSIONS: Although there were only minor differences in the physiological response of young and old bladders to obstruction, young rabbits showed a significantly greater degree of histological damage. This may have been due to the thinner wall and greater sensitivity to distention. PMID- 15758812 TI - Effects of saw palmetto extract on micturition reflex of rats and its autonomic receptor binding activity. AB - PURPOSE: We examined the effects of saw palmetto extract (SPE) on the rat micturition reflex and on autonomic receptors in the lower urinary tract. MATERIALS AND METHODS: The effect of SPE was examined on cystometrograms of anesthetized rats induced by intravesical infusion of saline or 0.1% acetic acid. SHR/NDmc-cp (cp/cp) rats received repeat oral administration of SPE and nighttime urodynamic function was determined. The autonomic receptor binding activity of SPE in the rat bladder and prostate was examined by radioligand binding assay. RESULTS: Intraduodenal administration of SPE (60 mg/kg) in anesthetized rat cystometry caused a significant increase in the micturition interval, micturition volume and bladder capacity during intravesical saline infusion. Also, similar administration of SPE at doses of 12 and 20 mg/kg significantly reversed the shortened micturition interval as well as the decreased micturition volume and bladder capacity due to 0.1% acetic acid infusion in a dose dependent manner. In conscious SHR/NDmc-cp (cp/cp) rats repeat oral administration of SPE (6 mg/kg daily) constantly increased the micturition interval and concomitantly decreased voiding frequency. SPE inhibited specific binding of [H]NMS ([N-methyl H]scopolamine methyl chloride) (bladder) and [H]prazosin (prostate) with IC50 values of 46.1 and 183 microg/ml, respectively. CONCLUSIONS: SPE significantly alleviates urodynamic symptoms in hyperactive rat bladders by increasing bladder capacity and subsequently prolonging the micturition interval. Our data may support the clinical efficacy of SPE for the treatment of lower urinary tract symptoms. PMID- 15758815 TI - Microfilaments are involved in renal cell responses to sustained hydrostatic pressure. AB - PURPOSE: Increased pressures within renal interstitial fluid, as associated with a number of renal pathologies, could affect cell function and gene expression. The long-term objective of this research is to elucidate kidney cell responses to pathological hydrostatic pressures. MATERIALS AND METHODS: In vitro studies were performed in 2 kidney cell lines (cortical tubular and medullary) to determine changes in cell numbers and cytoskeletal (specifically microfilament, microtubule and intermediate filament) arrangement following exposure to pathological (60 cm H2O) pressures. A novel pressure system was used to apply pressure to renal cells for up to 7 days. Cell counts and fluorescent staining were performed to determine alterations in response to pressure. RESULTS: Exposure to pressures of 60 cm H2O resulted in increased renal cell numbers and rearrangement in individual microfilament structures after 7 days. CONCLUSIONS: These results prove that hydrostatic pressure alters the function of renal cells. In the future such knowledge of renal cell responses to pressure along with an understanding of the mechanisms involved will aid in the design of novel, targeted drug therapies for treating kidney pathologies. PMID- 15758814 TI - Proceedings of the Baltimore smooth muscle meeting: identifying research frontiers and priorities for the lower urinary tract. AB - PURPOSE: The myocyte is a major parenchymal cell of the lower urinary tract (LUT) in men and women. Significant phenotypic diversity ensures that myocytes subserve their important role in the physiologically distinct tissues and organs of the LUT, including the ureters, bladder, urethra, prostate, penis, vagina and myometrium. Coordinated contraction and relaxation of myocytes is required for normal organ function, while alterations in myocyte structure/function are implicated in the etiology of various LUT diseases/disorders. LUT diseases/disorders will continue to increase in an ever aging American population. The purpose of the Baltimore Smooth Muscle Meeting was to begin to identify some research frontiers and priorities. MATERIALS AND METHODS: A 1-day conference of some of the leading world experts in smooth muscle research was held at American Urological Association headquarters. These experts gave presentations in their areas of expertise and extensively discussed their work. This report details those interactions. RESULTS: There is astonishing diversity in the contribution of the myocyte to LUT physiology and dysfunction. Novel tools, technologies and ideas have produced increased understanding and identified new frontiers. CONCLUSIONS: An improved understanding of urogenital myocyte physiology, function and dysfunction is required better to elucidate disease mechanisms and develop novel therapeutics. The First Annual Baltimore Smooth Muscle Meeting provided the first step in this direction. More coordinated LUT myocyte funding initiatives, the further development of research resources, tools and technologies, and exploration of the urogenital system as a model system for studying systems biology and integrative physiology are among the highest research priorities. PMID- 15758816 TI - Decreases in corporeal vascular endothelial growth factor expression precede vasoreactivity changes in cholesterol fed rabbits. AB - PURPOSE: We determined temporal changes in vasoreactivity and angiogenic growth factor levels in corporeal tissue at varying time points after the induction of hypercholesterolemia in rabbits. MATERIALS AND METHODS: A total of 42 New Zealand White rabbits were fed a 1% cholesterol (8 per group) or normal (6 per group) diet for 2, 4.5 or 7.5 weeks. Vascular endothelial growth factor (VEGF) mRNA expression in corpus cavernosum was assessed by real-time polymerase chain reaction analyses for the 3 isoforms VEGF121, VEGF165 and VEGF189. Isometric tension studies were performed and dose response curves were generated to evaluate endothelial dependent and endothelial independent vasoreactivity. RESULTS: Real-time polymerase chain reaction analysis showed 2.2 to 2.5 and 1.5 to 2.7-fold decreases in VEGF121 and VEGF165, respectively, in the corporeal tissues of the high cholesterol group vs the normal diet group at the 2 week time point. At 2 weeks VEGF189 was unchanged but it was decreased 1.5 to 2-fold at 4.5 weeks. Acetylcholine isometric tension studies revealed no difference in mean ED50 (-log [M]) +/- SD until 7.5 weeks of high cholesterol diet (5.10 +/- 0.64 vs 3.95 +/- 1.35, p = 0.0269). The response to sodium nitroprusside was not statistically different at any time point. Endothelial cell and smooth muscle content were decreased for the high cholesterol vs normal diet at 4.5 weeks (endothelial only) and 7.5 weeks (each cell). CONCLUSIONS: Alterations in corporeal tissue levels of VEGF occur before abnormalities in vasoreactivity. The results suggest that VEGF has a role in normal vasoreactivity in corporeal tissue and, thereby, in normal erectile function. PMID- 15758818 TI - Genomic Structure of Human OKL38 Gene and its Differential Expression in Kidney Carcinogenesis. PMID- 15758819 TI - Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine, and serum DNA of patients with renal cancer. PMID- 15758820 TI - Donor kidney exchanges. PMID- 15758817 TI - Effect of basic fibroblast growth factor incorporating gelatin microspheres on erectile function in the diabetic rat. AB - PURPOSE: We report the potential of basic fibroblast growth factor (bFGF) incorporating gelatin microspheres to preserve erectile function in a diabetic rat model. MATERIALS AND METHODS: A total of 48 adult male rats were divided into 3 groups, namely control (nondiabetic rats), diabetes mellitus (DM) (diabetic rats that received gelatin microspheres with saline) and bFGF (diabetic rats that received gelatin microspheres with bFGF). After 4 and 8 weeks we examined intracavernous pressure responses with electrical stimulation to the cavernous nerve. For histological examination of the penis we performed Azan-Mallory staining for smooth muscle and collagen, and immunohistochemistry for endothelial nitric oxide synthase (NOS) in endothelium and neuronal NOS in cavernous nerve fiber. RESULTS: Although the intracavernous pressure response was significantly lower in the DM group than in the control group, pressure in the bFGF group was maintained at the normal level found in controls. Azan-Mallory staining showed a mass decrease in smooth muscle in cavernous tissue in the DM group. However, that in the bFGF group was maintained. There was no significant difference in endothelial NOS positive areas and the distribution of the diameter of neuronal NOS positive nerve fibers in cavernous tissue among the 3 groups. CONCLUSIONS: We report the maintenance of erectile function with bFGF incorporating gelatin microspheres in diabetic rats. The rationale of this maneuver is smooth muscle preservation by the long-term release of bFGF. This is a novel therapeutic option that is clinically applicable for diabetes induced erectile dysfunction. PMID- 15758821 TI - Amitriptyline eliminates calculi through urinary tract smooth muscle relaxation. PMID- 15758823 TI - Xenotransplantation and the future of renal replacement. PMID- 15758822 TI - Tissue engineering stem cells, and cloning: opportunities for regenerative medicine. PMID- 15758824 TI - Autonomic dysfunction in Parkinson's disease. PMID- 15758826 TI - Re: Is tumor volume an independent prognostic factor in clinically localized prostate cancer? PMID- 15758827 TI - Re: A single immediate postoperative instillation of chemotherapy decreases the risk of recurrence in patients with stage Ta T1 bladder cancer: a meta-analysis of published results of randomized clinical trials. PMID- 15758829 TI - RE: Effect of the dual 5alpha-reductase inhibitor dutasteride on markers of tumor regression in prostate cancer. PMID- 15758830 TI - Re: Vena caval transection during retroperitoneoscopic nephrectomy: report of the complication and review of the literature. PMID- 15758831 TI - Re: Urinary conduit formation using a retubularized bowel from continent urinary diversion or intestinal augmentations: II. Does it have a role in patients with interstitial cystitis? PMID- 15758833 TI - Re: Functional and neuroanatomical effects of vaginal distention and pudendal nerve crush in the female rat. PMID- 15758836 TI - Sarcoidosis in patients with chronic hepatitis C virus infection: analysis of 68 cases. AB - We describe the clinical characteristics, the patterns of association, and the role of antiviral therapies in patients with sarcoidosis associated with chronic hepatitis C virus (HCV) infection. Sixty-eight patients were included in the current study, 56 cases identified in the literature search plus 12 unpublished cases from our department. In 50 HCV patients, sarcoidosis appeared after starting antiviral therapy. Antiviral therapy associated with triggered sarcoidosis consisted of alpha-interferon monotherapy in 20 cases and combined therapy with alpha-interferon and ribavirin in 30. Sarcoidosis appeared during the first 6 months after starting therapy in 66% of patients. The clinical picture of sarcoidosis included predominantly pulmonary disease in 38 (76%) patients and cutaneous sarcoidosis in 30 (60%). Antiviral therapy was discontinued in 60% of patients and continued or adjusted in 14%, while sarcoidosis appeared after completed therapy in the remaining cases. Specific therapy for sarcoidosis was started in only 21 patients, mainly with oral corticosteroids. The outcome of patients was detailed in 46 cases: remission or improvement was observed in 38/46 (83%) patients, stabilization of sarcoidosis in 5/46 (11%), and reactivation of sarcoidosis after an initial improvement in 3/46 (6%). Finally, 18 treatment-naive HCV patients presented sarcoidosis, with 14/18 (87%) patients presenting with pulmonary involvement and 8/18 (44%) with cutaneous involvement. In summary, sarcoidosis may be observed in HCV patients in 2 different situations: triggered by antiviral therapy (in 75% of cases) and unrelated to treatment. Sarcoidosis during antiviral therapy may present mainly as cutaneous or pulmonary disease, with a benign, uncomplicated evolution in more than 85% of cases. However, more complicated cases are observed, especially in HCV patients with preexisting sarcoidosis and/or with previous antiviral treatment. Clinicians should be aware of the possibility that sarcoidosis may initially manifest or be reactivated during or shortly after treatment with antiviral therapy in patients with chronic HCV infection. PMID- 15758837 TI - Sjogren syndrome associated with hepatitis C virus: a multicenter analysis of 137 cases. AB - To define the clinical and immunologic pattern of expression of Sjogren syndrome (SS) associated with chronic hepatitis C virus (HCV) infection, we conducted a multicenter study aiming to collect a large number of patients with SS and HCV infection. Inclusion criteria were the fulfillment of at least 4 of the classification criteria for SS proposed by the European Community Study Group and repeated positive HCV serology, confirmed by recombinant immunoblot assay and/or detection of serum HCV-RNA by polymerase chain reaction. One hundred thirty-seven patients were included (104 female and 33 male; mean age, 65 yr). Seventy-nine (58%) patients presented a systemic process with diverse extraglandular manifestations, with articular involvement (44%), vasculitis (20%), and neuropathy (16%) being the most frequent features observed. The main immunologic features were antinuclear antibodies (65%), hypocomplementemia (51%), and cryoglobulinemia (50%). Cryoglobulins were associated with a higher frequency of cutaneous vasculitis, rheumatoid factor, and hypocomplementemia. Thirty-two (23%) patients had positive anti-Ro/SS-A and/or anti-La/SS-B antibodies; these patients were predominantly women and had a higher prevalence of some extraglandular features and a lower frequency of liver involvement. Nineteen (14%) patients developed neoplasia, with hematologic neoplasia (8 cases) and hepatocellular carcinoma (6 cases) being the most frequent types. Eighty-five percent of SS-HCV patients also fulfilled the recently proposed 2002 classification criteria for SS. In conclusion, HCV-associated SS is indistinguishable in most cases from the primary form using the most recent set of classification criteria. Chronic HCV infection should be considered an exclusion criterion for the classification of primary SS, not because it mimics primary SS, but because the virus may be implicated in the development of SS in a specific subset of patients. We propose the term "SS secondary to HCV" when these patients fulfill the 2002 classification criteria for SS. PMID- 15758838 TI - Circulating monoclonal immunoglobulins in Sjogren syndrome: prevalence and clinical significance in 237 patients. AB - We conducted the current study to analyze the prevalence and clinical significance of circulating monoclonal immunoglobulins in patients with Sjogren syndrome (SS), focusing on the association with extraglandular features, immunologic markers, hematologic neoplasia, and hepatitis C virus (HCV) infection. We performed serum immunoelectrophoresis in 200 patients with primary SS and 37 patients with HCV-related SS. All patients fulfilled 4 or more of the 1993 European classification criteria for SS.Of the 200 patients with primary SS, 35 (18%) presented circulating monoclonal immunoglobulins. The monoclonal bands identified were 20 IgG (13 kappa, 7 lambda), 10 IgM (5 kappa, 5 lambda), 2 IgAkappa, and 3 free circulating light chains. Of the 37 SS-HCV patients, 16 (43%) had circulating monoclonal immunoglobulins. The monoclonal bands identified were 10 IgMkappa, 5 IgGlambda, and 1 free light lambda chain. Compared with primary SS patients, SS-HCV patients presented a higher frequency of monoclonal immunoglobulins (43% vs 18%, p = 0.001), with monoclonal IgMkappa being the most frequent monoclonal band. Six (12%) of the 51 SS patients with circulating monoclonal immunoglobulins presented hematologic neoplasia, compared with 3 (1.6%) of those without monoclonal immunoglobulins (p = 0.004; odds ratio = 8.13; 95% confidence intervals, 1.64-51.54). In 2 of the 6 patients with monoclonal immunoglobulins and lymphoproliferative disorders, a change of the monoclonal component was detected in previous immunoelectrophoresis determinations before the development of hematologic neoplasia. Circulating monoclonal immunoglobulins were detected in nearly 20% of patients with primary SS, with monoclonal IgG being the most frequent type of immunoglobulin detected. In SS-HCV patients, the prevalence of monoclonal immunoglobulins was higher (43%), with monoclonal IgM being the most frequent type found. SS-HCV patients presented a more restrictive monoclonal expression (limited to either monoclonal IgMkappa or monoclonal IgGlambda) than primary SS patients, who showed all types of heavy and light chains. PMID- 15758839 TI - Characteristics of autoimmune thyroid disease occurring as a late complication of immune reconstitution in patients with advanced human immunodeficiency virus (HIV) disease. AB - Experimental evidence from animal models has provided a framework for our current understanding of autoimmune disease pathogenesis and supports the importance of genetic predisposition, molecular mimicry, and immune dysregulation. However, only recently has evidence emerged to support the role of immune dysregulation in human organ-specific autoimmune disease. In the current study of the "late" manifestation of autoimmune thyroid disease (AITD) in a cohort of human immunodeficiency virus (HIV)-positive patients following highly active antiretroviral therapy (HAART), we discuss how immune dysregulation and factors associated with the immunopathology of HIV infection fit the current understanding of autoimmunity and provide a plausible basis for our clinical observations. De novo diagnoses of thyroid disease were identified between 1996 and 2002 in 7 HIV treatment centers (5/7 centers completed the study). Patients were diagnosed as clinical case entities and not discovered through thyroid function test screening. Paired plasma specimens were used to demonstrate sequential rise in thyroid antibodies. Seventeen patients were diagnosed with AITD (median age, 38 yr; 65% were of black African or black Caribbean ethnicity; and 82% were female). The median duration of immune reconstitution was 17 months. Graves disease (GD) was diagnosed in 15 of 17 patients. One patient developed hashithyrotoxicosis with atypically raised C-reactive protein, and another developed hypothyroidism. One GD patient had associated secondary hypoadrenalism. The estimated combined prevalence of GD for 4 treatment centers for female patients was 7/234 and for males was 2/1289. The denominator numbers were matched controls, from 4 centers able to provide data, who commenced HAART during the same time (January 1996 to July 2002) and who did not develop clinical AITD. The mean baseline pre-HAART CD4 count was 67 cells/mL, and the mean increase from nadir to AITD presentation was 355 cells/mL. AITD patients were more likely than controls (95% confidence interval, chi-square test) to be severely compromised at baseline (as defined by a CD4 count < 200 cells/mL or the presence of an acquired immunodeficiency syndrome [AIDS]-defining diagnosis), and to experience greater CD4 increments following HAART. AITD may be a late manifestation of immune reconstitution in HIV-positive patients taking HAART, and immune dysregulation may be an important factor. PMID- 15758840 TI - Comparison of 3 clinical models for predicting the probability of pulmonary embolism. AB - Two clinical models have been described to predict the probability of pulmonary embolism: the Canadian (or Wells) model, and the Geneva model. A third model has been developed recently at our institution (the Pisa model). We compared the performance of the 3 models in 215 consecutive patients with suspected pulmonary embolism. The clinical probability predicted by the models was categorized as low, intermediate, or high. In all patients, pulmonary angiography was used as the reference diagnostic standard. In patients with pulmonary embolism, the extent of pulmonary embolization was assessed on the lung scan as an index of disease severity. The prevalence of pulmonary embolism was 43.3%, and the median extent of pulmonary embolization at diagnosis was 39.8% (range, 4.5%-75.3%). The proportions of patients categorized as having low, intermediate, or high probability were, respectively: 12%, 60%, and 28%, for the Geneva model; 30%, 55%, and 15%, for the Wells model; 37%, 37%, and 26% for the Pisa model. The frequencies of pulmonary embolism in the low, intermediate, and high probability categories were, respectively: 50%, 39%, and 49% for the Geneva model; 12%, 54%, and 64% for the Wells model; 5%, 42%, and 98% for the Pisa model. Among patients with pulmonary embolism, there was a strong, positive relation between clinical probability predicted by the Pisa model and the extent of pulmonary embolization. The Pisa model proved more accurate than the 2 other models. It may be useful to physicians in defining precisely the pretest probability of pulmonary embolism. PMID- 15758841 TI - Presentation and outcome of gastrointestinal involvement in systemic necrotizing vasculitides: analysis of 62 patients with polyarteritis nodosa, microscopic polyangiitis, Wegener granulomatosis, Churg-Strauss syndrome, or rheumatoid arthritis-associated vasculitis. AB - We reviewed the medical records of 62 patients with systemic small and medium sized vessel vasculitides and gastrointestinal tract involvement followed at our institution between 1981 and 2002. This group included 46 men and 16 women (male:female ratio, 2.9), with a mean age of 48 +/- 18 years. Vasculitides were distributed as follows: 38 polyarteritis nodosa (21 related to hepatitis B virus), 11 Churg-Strauss syndrome, 6 Wegener granulomatosis, 4 microscopic polyangiitis, and 3 rheumatoid arthritis-associated vasculitis. Gastrointestinal manifestations were present at or occurred within 3 months of diagnosis in 50 (81%) patients and were mainly abdominal pain in 61 (97%), nausea or vomiting in 21 (34%), diarrhea in 17 (27%), hematochezia or melena in 10 (16%), and hematemesis in 4 (6%). Gastroduodenal ulcerations were detected endoscopically in 17 (27 %) patients, esophageal in 7 (11%), and colorectal in 6 (10%), but histologic signs of vasculitis were found in only 3 colon biopsies. Twenty-one (34%) patients had a surgical abdomen; 11 (18%) developed peritonitis, 9 (15%) had bowel perforations, 10 (16%) bowel ischemia/infarction, 4 (6%) intestinal occlusion, 6 (10%) acute appendicitis, 5 (8%) cholecystitis, and 3 (5%) acute pancreatitis. (Some patients had more than 1 condition.) Sixteen (26%) patients died.The respective 10-month and 5-year survival rates were 71% (95% confidence interval [CI], 52-90) and 56% (95% CI, 35-77) for the 21 surgical patients; and 94% (95% CI, 87-101) and 82% (95% CI, 70-94) for the 41 patients without surgical abdomen (p = 0.08). Peritonitis (hazard ratio [HR] = 4.3, p < 0.01), bowel perforations (HR = 5.7, p < 0.01), gastrointestinal ischemia or infarctions (HR = 4.1, p < 0.01), and intestinal occlusion (HR = 5.5, p < 0.01) were the only gastrointestinal manifestations significantly associated with increased mortality in multivariate analysis. For this subgroup of 15 patients, 6-month and 5-year survival rates were 60% (95% CI, 35-85) and 46% (95% CI, 19-73), respectively (p = 0.003). None of the other gastrointestinal or extraintestinal vasculitis related symptoms, or angiographic abnormalities (seen in 67% of the 39 patients who underwent angiography), was predictive of surgical complications or poor outcome. However, prognosis has dramatically improved during the past 30 years, probably owing to better management of these more severely ill patients, with prompt surgical intervention when indicated, and the combined use of steroids and immunosuppressants. PMID- 15758842 TI - Brain magnetic resonance imaging in patients with Cowden syndrome. AB - Cowden syndrome (CS) is a rare autosomal dominant genodermatosis, characterized by multiple hamartomas, particularly of the skin, associated with high frequencies of breast, thyroid, and genitourinary malignancies. Although Lhermitte-Duclos disease (LDD) or dysplastic gangliocytoma of the cerebellum, a slowly progressive unilateral tumor, is a major criterion of CS, its frequency in patients with CS is unknown. Other cerebral abnormalities, especially meningioma and vascular malformations, have also been described, albeit rarely, in these patients. The aim of the current study was to use cerebral magnetic resonance imaging (MRI) to evaluate LDD frequency and to investigate other brain abnormalities in CS patients recruited by dermatologists. A multicenter study was conducted in 8 hospital dermatology departments between January 2000 and December 2003. Twenty patients with CS were included; specific cerebral MRI abnormalities were found in 35% (7/20) of them. Cerebral MRI revealed LDD in 3 patients, a meningioma in 1, and numerous vascular malformations in 6 patients. Five patients had venous angiomas (3 associated with LDD) and 2 patients had cavernous angiomas (1 associated with LDD and a venous angioma). The discovery of asymptomatic LDD in 3 patients and a cavernous angioma in another prompted us to perform neurologic examinations regularly and MRI to estimate the size and the extension of the tumor, and to assess the need for surgery. CS similarities with Bannayan Riley-Ruvalcaba (BRR) are discussed because some patients could also have the BRR phenotype (for example, genital lentigines, macrocephaly, multiple lipomas) and because BRR seems to have more central nervous system vascular anomalies. Because CS signs can involve numerous systems, all physicians who might encounter this disease should be aware of its neurologic manifestations. Our findings confirm the contribution of brain MRI to detecting asymptomatic LDD, vascular malformations, and meningiomas in patients with CS. PMID- 15758843 TI - Laboratory blood lactate profile is suited to on water training monitoring in highly trained rowers. AB - AIM: This study validated the laboratory testing used to monitor on water training. The purpose was to test that reference heart rates (HR) determined during an incremental test elicit comparable blood lactate levels ([La](b)) during a 30 min on water rowing. METHODS: Blood lactate profile were determined during incremental graded exercise in 14 national and international level oarsmen. The HR corresponding to [La](b) of 2 and 3 mmol x l(-1) were determined (HRLa2 and HRLa3 respectively). The rowers then performed a 30 min training session in a boat. Training intensity, as assessed by HR monitors, had to range between HRLa2 and HRLa3. Field [La](b) (Laf) and HR (HRf) were measured at the end of the training session. RESULTS: Laf was 2.13+/-0.49 mmol x l(-1) (range: 1.43-3.07) and did not differ significantly from 2 mmol x l(-1). HRf (162+/-7.4 beats x min(-1)) ranged from HRLa2 (159+/-9.5 beats x min(-1)) to HRLa3 (171+/-9 beats x min(-1)). HRf was not significantly different from HRLa2. CONCLUSIONS: It was concluded that the HR determined during the laboratory testing are valid for monitoring on water training in highly trained rowers. PMID- 15758845 TI - Application of force-velocity cycle ergometer test and vertical jump tests in the functional assessment of karate competitor. AB - AIM: The aim of this study was to analyze the links between tests performances (vertical jump and force-velocity sprint on cycle ergometer) and 2 different karate level groups in order to propose a test battery adjusted to karate. METHODS: Twenty-two karate competitors (10 national junior team (IJ) and 12 national competition level (NL)) performed 4 maximal squat jumps (SJ), 4 maximal counter movement jumps (CMJ) on an ergojump and 3 8-s sprints on a friction braked cycle ergometer (friction loads of 0.5, 0.7, 0.9 N x kg(-1)). The maximal theoretical force (F(0)) and velocity (V(0)), the maximal power output (P(max)) and the optimal pedalling velocity (V(opt)) were derived from both the force -- velocity and the power -- velocity relationships plotted from all the 3 friction loads data. V(0), F(0), V(opt), P(max) and the best SJ and CMJ, were compared between IJ and NL groups. RESULTS: The IJ group was characterised by significantly higher values of V(0) (+13%) and SJ (+14.3%) compared to NL group, whereas no significant difference was observed between groups for F(0). Thus, karate performance would depend on maximal velocity and explosive strength. In addition, V(opt) was significantly higher in IJ group compared to NL group (135.4 rpm vs 119.2 rpm, p<0.001). Although based upon indirect evidence, these results accounted for mechanical functional capabilities of experts which could be particularly valuable when monitoring training of karate competitor. CONCLUSIONS: A force-velocity and a vertical jump tests may be applied in the functional assessment of karate competitor. PMID- 15758844 TI - Exercise induced arterial hypoxemia in swimmers. AB - AIM: Exercise induced arterial hypoxemia (EIAH) is a reduction in arterial oxygenation, which may result from a drop in arterial oxygen pressure and therefore in oxygen saturation. We examined EIAH in swimmers, while till now it was known to occur in cyclists and runners. METHODS: We studied 8 male highly trained swimmers (age: 23+/-1.7; (.-)VO(2peak), 5.3+/-0.1 l/min and 8 male ex swimmers (age: 21.5+/-0.6; (.-)VO(2peak), 3.4+/-0.3 l/min). All subjects performed 200-meter freestyle at maximum effort. Hemoglobin saturation (SaO(2)%) was measured using a finger pulse oximeter before exercise in the water in an upright position and immediately after exercise, within 5 seconds. RESULTS: Highly trained swimmers developed a statistically significant decrease in SaO(2)% (from 98.3+/-0.3 to 94+/-0.9, p= or <0.01) after exercise, while ex-swimmers did not (from 98.4+/-0.3 to 96.8+/-0.3 ns). The 4% decrease in SaO(2)% observed in highly trained swimmers can be characterized as mild EIAH. CONCLUSIONS: Our study suggests that highly trained swimmers but not ex-swimmers may develop mild EIAH after 200 meters freestyle swimming at maximum effort. PMID- 15758846 TI - Full suspension mountain bike improves off-road cycling performance. AB - AIM: The purpose of the present study was to determine the effects of suspension systems on the cycling performance of cyclists during off-road bicycling. METHODS: Eight elite male cyclists (67.8+/-5.8 ml/min/kg of (.-)VO(2max)) performed 30-minute riding tests on bicycles with 2 different suspension setups: front suspension (FS) and front and rear suspension (FRS). Heart rate, blood lactate concentration, pedaling power, cadence, cycling velocity, and completed distance during the trial were measured creatin kinase (CK), lactic dehydrogenase (LDH) and glutamic-oxaloacetic transaminase (GOT) were measured before and after the trials. RESULTS: The average cadence during the trial was significantly higher (p<0.05) with the FRS (73.6+/-6.1 rpm) than the FS (70.2+/-6.2 rpm). Subjects rode significantly faster (p<0.05) on FRS (24.1+/-2.6 km/h) than FS bikes (22.9+/-2.4 km/h), although no significant difference was observed in pedaling power (240.7+/-26.6 W vs 242.2+/-28.8 W, FS vs FRS, respectively). Serum creatin kinase increased significantly (p<0.05) at 24 h after the trial when cyclists exercised with the FS bike. CONCLUSIONS: We conclude that the FRS improved cycling performance over rough terrain. FRS might therefore be more suitable for cross-country mountain bike races. PMID- 15758847 TI - The effect of continuous repetition training and intra-set rest training on bench press strength and power. AB - AIM: The purpose of this research was to compare the effects of continuous repetition and intra-set rest training on maximal strength and power output of the upper body. METHODS: The 6 repetition maximum (6RM) and bench press throw power output against masses of 20, 30 and 40 kg of 26 elite junior male basketball and soccer players were tested on 2 separate occasions for reliability purposes. Subjects were then randomly assigned to either a continuous repetition (CR - 4 sets x 6 repetitions) or intra-set rest (ISR - 8 sets x 3 repetitions) training regime over 6-weeks. Volume (sets x repetitions x %6RM) between groups was equated and both groups completed all sets in the same time period (13 minutes and 20 seconds). The total concentric work time was determined to identify differences in training regimes. Independent sample t-tests on preintervention and postintervention percentage change scores were analysed for significant differences (p<0.05). RESULTS: The observed coefficients of variation (1.7% to 4.8%) and intraclass correlation coefficients (r=0.87 to 0.98) indicated stability of these measures across testing occasions. The CR group significantly increased 6RM strength (9.7%) compared with the ISR group (4.9%). The total concentric work time was significantly longer in CR training than ISR (36.03+/- 4.03 s and 31.74+/-4.71 s; p=0.13). Power output increases across the 20, 30 and 40 kg loads ranged from 5.8% to 10.9% for both training groups but the between group percentage change scores were not significantly different. CONCLUSIONS: Bench press training involving 4 sets of 6 continuous repetitions elicited a greater improvement in bench press strength than 8 sets of 3 repetitions at the same percentage load of their 6RM. Both ISR and CR training were equally effective in increasing power output. PMID- 15758848 TI - Effect of long duration exercise on the ratings of perceived exertion and perceived difficulty of walking and running at the ventilatory threshold. AB - AIM: The purpose of this study was to examine if the relationship between physiological changes classically observed with exercise duration and some subjective workload measures would be affected by the complexity of the locomotion mode (running vs racewalking). METHODS: The study was conducted on 24 well trained subjects (12 long distance runners and 12 racewalkers) divided in 3 groups (runners, racewalkers and control). Energy cost of locomotion (C), heart rate (HR), minute ventilation (V(E)), lactate concentration [La], ratings of perceived exertion (RPE 6-20 scale) and ratings of perceived difficulty (RPD 1( 1)5 scale) were recorded during 2 10-min submaximal tests on a treadmill before and immediately after a 3 hour exercise (racewalking or running) conducted at the velocity ventilatory threshold (vVT). RESULTS: No significant variations in physiological parameters and perceived measures were observed in G(c). A significant increase (p<0.05) in energy cost of walking (mean: +9.4%) and running (mean: +7.5%) at the end of exercise was observed. A significant interaction of locomotion mode and exercise duration was found on perceived exertion and perceived difficulty. In racewalkers RPD significantly increased with duration whereas no significant effect was found for RPE. Conversely a significant increase in RPE was found after 3 hours in runners without any significant change in RPD. CONCLUSIONS: This experiment suggests that, for a complex task, the classical relationship between RPE and metabolic load increase during prolonged exercise could be affected by changes in RPD. In this study, stability in RPE and increase in RPD observed in racewalkers may reflect an attentional focus dissociated from internal sensations and directed toward maintaining the required race walking gait. PMID- 15758849 TI - Assessment of maximal aerobic velocity in soccer players by means of an adapted Probst field test. AB - AIM: The aim of the study was to test the accuracy of 2 versions of a specific soccer field test for assessing maximal aerobic velocity (MAV) in soccer players. The original Probst field test consists of repetitions of 280 m runs including changes in direction separated by a 30-second rest with an initial speed of 8.4 km x h(-1) and a 0.6 km x h(-1) increment at each stage. The adapted version was carried out with the same protocol but constant stages of 2-minute durations and a 1.2 km x h(-1) increment at each stage. METHODS: Trained soccer players completed 4 outdoor field tests: the original and adapted field Probst tests, both with and without changes in direction, in order to determine the velocities at exhaustion. They all underwent at the same time 3 laboratory tests in order to determine MAV during a classical graded treadmill test, a laboratory test with the same stages as the original Probst test and the adapted Probst test. RESULTS: The velocities at exhaustion with and without changes in direction obtained during the adapted Probst field test, and the MAV obtained in the laboratory were highly correlated. All were significantly lower than the velocities at exhaustion obtained during the original version. CONCLUSIONS: The adapted version was a better predictor of MAV in trained soccer players than was the original test. PMID- 15758850 TI - Running performance in middle-school runners. AB - AIM: This study examined the relationship of 3-km run time to indices of aerobic and anaerobic ability in 9 male runners (13.4+/-0.6 years, mean+/-SD). METHODS: Anthropometric measurements were made, and an exercise test to determine running economy at 187 m x min(-1) and (.-)VO(2max) were assessed on a treadmill. On a separate day, 2 55-m sprints followed by a 3-km run were performed on a 200-m indoor track. Capillary blood samples were obtained from a finger tip immediately after the run to determine blood lactate level. Fractional utilization (%(. )VO(2max) used during the 3-km run) was calculated. Correlations were used to examine the relationship between run time and the physiological measurements. RESULTS: Mean values for (.-)VO(2), HR and RER at maximal exercise were 61.7+/ 4.4 ml x kg(-1)xmin(-1), 198.9+/-6.7 b x min(-1), and 1.16+/-0.04, respectively. The average time to run 3 km was 13.27+/-0.97 min (90.1+/-7.2% of (.-)VO(2max)). Post-run blood lactate level was 8.3+/-3.2 mmol x L(-1) and was significantly related (r=-0.73, p=0.02) to 3-km time. Fractional utilization tended to be related (r=-0.56, p=0.12) to time. CONCLUSIONS: In this age group the ability to run at a high percentage of (.-)VO(2max) and tolerate a high blood lactate appear to be important determinants of running performance in young male runners. PMID- 15758851 TI - Combined effects of age and maturation on maximum isometric leg press strength in young basketball players. AB - AIM: Muscular strength of the leg extensor muscles in children can be affected by several factors such as age, sexual maturation, body mass and training status of the subjects. The purpose of the study was to examine maximal isometric strength characteristics of young male basketball players taking into consideration the combined effects of chronological age and sexual maturation. METHODS: One hundred and twenty male basketball players, aged from 12 to 17 years divided into 6 equivalent age subgroups performed maximum bilateral isometric leg press efforts. The parameters analysed were the maximal voluntary isometric force (MVC), relative strength (MVC/body mass and MVC/fat free mass), starting strength (F50: force exerted during the first 50 ms of the contraction) and speed strength index (the ratio of maximal force to time to attain maximal force). RESULTS: The results indicated that in almost all absolute force parameters, the 12-and 13 year olds demonstrated significantly (p<0.05) lower values compared with the 15( 1)6-and 17-years old groups. Age differences were also significant (p<0.05) when the effects of sexual maturation were taken into consideration in the statistical analysis but they were reduced when strength was adjusted for body mass. Finally, no significant differences for strength per unit of fat free mass were found (p>0.05). CONCLUSIONS: Maximum absolute strength of basketball players is significantly increased from 12 to 17 years and as sexual maturation stage increases. It also appears that body mass and fat free mass should be taken into consideration when examining age effects on strength in basketball players. PMID- 15758852 TI - Energy expenditure, heart rate response, and metabolic equivalents (METs) of adults taking part in children's games. AB - AIM: The needs of physical activity can be seen through the lack of numbers participating in regular physical activity as well as the increase in prevalence of certain diseases such as Type II diabetes (especially in children), cardiovascular diseases, and some cancers. With the increase in preventable diseases that are caused in part by a sedentary lifestyle, a closer look needs to be taken into the role of family interaction as a means of increasing physical activity for both adults and children. Because of the many benefits of physical activity in relation to health, a family approach to achieving recommended levels of physical activity may be quite applicable. METHODS: Forty volunteers were recruited from the community (20 subjects and 20 children). The volunteers played 2 games: soccer and nerfball. Data was collected over 10 minutes (5 min per game). Expired air analysis was used to calculate energy expenditure and metabolic equivalents (METs). Descriptive statistics were calculated along with a regression analysis to determine differences between the 2 games, and an ACOVA to determine any significant effects of age, child age, gender, and physical activity level on the results. RESULTS: For both games, average heart rate measured approximately 88%max; average METs measured approximately 6, average energy expenditure measured approximately 40 kcal. CONCLUSIONS: S: This study showed that adults can achieve recommended physical activity levels through these specific activities if sustained for approximately 20 min. PMID- 15758853 TI - Does aerobic training improve mental stress tolerance in job situations? AB - AIM: Effects of aerobic fitness on cognitive performance (CP) in elderly persons have been investigated with contradictory RESULTS: However, these effects have not been explored in occupationally active young and healthy subjects yet. Therefore we studied the effects of an aerobic training on CP during mental stress and in low demanding situations. METHODS: Twenty-seven healthy volunteer students performed a maximal bicycle exercise test and 2 computerized psychological examinations at the begin and the end of a 3 month period of individualized aerobic training. The psychological tests were the ''Determinations-test'' (DT), which is a multi-task stress test, and the ''Vigilance test'' (VT), representing a low demanding situation. According to the outcome of the aerobic training 2 groups were formed: those with moderate and those with good training effects. RESULTS: Results showed no significant differences between the 2 groups over the training period (5 parameters at DT and 4 at VT were n.s., 1 result at VT with p<0.05 was interpreted as a regression to the mean). CONCLUSIONS: We conclude that neither CP at high stress nor vigilance during low demanding periods can be influenced by an improvement of aerobic fitness in occupationally active young and healthy persons. PMID- 15758854 TI - Is the use of oral creatine supplementation safe? AB - This review focuses on the potential side effects caused by oral creatine supplementation on gastrointestinal, cardiovascular, musculoskeletal, renal and liver functions. No strong evidence linking creatine supplementation to deterioration of these functions has been found. In fact, most reports on side effects, such as muscle cramping, gastrointestinal symptoms, changes in renal and hepatic laboratory values, remain anecdotal because the case studies do not represent well-controlled trials, so no causal relationship between creatine supplementation and these side-effects has yet been established. The only documented side effect is an increase in body mass. Furthermore, a possibly unexpected outcome related to creatine monohydrate ingestion is the amount of contaminants present that may be generated during the industrial production. Recently, controlled studies made to integrate the existing knowledge based on anecdotal reports on the side effects of creatine have indicated that, in healthy subjects, oral supplementation with creatine, even with long-term dosage, may be considered an effective and safe ergogenic aid. However, athletes should be educated as to proper dosing or to take creatine under medical supervision. PMID- 15758855 TI - Association of habitual long-distance running with the thickness of skeletal muscles and subcutaneous fat in the body extremities and trunk in middle-aged men. AB - AIM: We investigated the association of habitual long-distance running with the thickness of skeletal muscles and subcutaneous fat in the body extremities and trunk in middle-aged men using ultrasonography. METHODS: Three groups of healthy middle-aged men [mean (SD), 62.1 (2.8) years] took part in this investigation: a high-level group of 11 master runners who had competed in a 42.195 km race and run 51.6 (21.7) km every week, an intermediary-level group of 10 master runners who had competed in a 5-20 km race and run 9.3 (4.9) km every week, and a low level group of 7 untrained men who continued to do no systematic training. The muscle thickness at 8 sites and the subcutaneous fat thickness at 10 sites were measured by B-mode ultrasonography, and were compared among the 3 groups. RESULTS: The high-level group had 10.0(-1)5.2% higher values for muscle thickness at the erector spinae, hamstrings, tibialis anterior, and triceps surae, compared with the intermediary-level and the low-level groups (p<0.05-0.001). The thickness of the subcutaneous fat about the rectus abdominis and external oblique was lower in the high-level group than in the intermediary-level and the low level groups (p<0.05). CONCLUSIONS: Middle-aged male master athletes who habitually run at a high level have more muscle thickness in the lower extremities and trunk, and less subcutaneous fat thickness in the central regions of the body than do middle-aged men who habitually run at an intermediary level or do not run at all. PMID- 15758856 TI - Weight lifting and type II aortic dissection. A case report. AB - The case of a 28-year-old previously healthy male with a 13-year history of weight lifting is described. The patient presented to the emergency department complaining of severe anterior chest pain beginning during a workout. Following transthoracic echocardiogram and computerized tomogram the diagnosis of an ascending aortic dissection was made. The patient was taken emergently for surgical repair of an incompetent aortic valve and type II aortic dissection. Histopathologic examination of the aorta showed cystic medial degeneration. This is the first reported case of a type II aortic dissection secondary to cystic medial degeneration and the hemodynamic stresses of weight lifting. Physicians should be aware of this potentially catastrophic entity in weight lifters. PMID- 15758857 TI - Development of the Muscle Dysmorphia Inventory (MDI). AB - AIM: The development of the 6-factor, 27-item Muscle Dysmorphia Inventory (MDI) was based on Lantz et al. proposed model of characteristics associated with Muscle Dysmorphia. METHODS: EXPERIMENTAL DESIGN: quantitative procedures including item-to-total correlations, exploratory and confirmatory factor analyses, and structure equation modeling confirmed the construct validity of the scale. Convergent validity was also tested. SETTING: bodybuilding and powerlifting competition venues, weight training facilities, and university athletic venues. PARTICIPANTS: the 1(st) study consisted of 77 experienced male free weight lifters. The 2(nd) study consisted of 156 male non-competitive bodybuilders and weight lifters and 168 elite level powerlifters and bodybuilders. The 3(rd) study consisted of 151 male and female bodybuilders and weight lifters. MEASURES: each participant completed demographic information, the MDI, Drive for Thinness subscale of the Eating Disorder Inventory, and the Training Dependency subscale of the Bodybuilding Dependence Scale. RESULTS: Reliability estimates (Cronbach's a) ranged from 0.72 to 0.94. Factor loadings in all 3 studies supported the 6-factor structure (size/symmetry, supplement use, exercise dependence, pharmacological use, dietary behavior, and physique protection). Much of the scale validation was focused on construct validity, however, correlations with the MDI's subscales and the Training Dependency subscale of the Bodybuilding Dependence Scale and the Drive for Thinness subscale of the Eating Disorder Inventory provided evidence of convergent validity also. CONCLUSIONS: From these preliminary results, the MDI appears to contribute to the identification of a newly formed disorder by offering a multi-dimensional measure of factors related to Muscle Dysmorphia. PMID- 15758858 TI - Deep peroneal nerve paresis in a runner caused by ganglion at capitulum peronei. Case report and review of the literature. AB - Although lateral popliteal sciatic nerve damage is not one of the commonest diseases in the general population, it is quite frequent among athletes. Several physiopathologic mechanisms have been thought to bring about this damage in athletes. Soft tissue ganglions with neurological involvement of the lateral popliteal sciatic nerve or its terminal rami are in differential diagnosis with several lesions of this area, as direct or indirect trauma, subcutaneous rupture of anterior tibialis muscle and long peroneal muscle, disc hernia, intraspinal tumor, anterior tarsal tunnel syndrome, cysts, neurofibroma, baker's cyst, vascular claudication, stenosing or inflammatory pathology of 2(nd) motoneuron, antimicrobial agents for urinary tract infection (nitrofurnantoin). The authors report the case of a 34-year-old amateur athlete with a recent paralysis of the hallux extensor, paresis of the toe extensor and hyposthenia of the tibialis anterior. The patient had been suffering from episodes of lumbalgia for a long time. He was sent to us because neurological damage due to disc herniation was suspected. Electromyography, sonography, and CT showed peripheral compression of the deep peroneal nerve caused by a mucous cyst at the capitulum peronei, a ''rare'' condition. The patient underwent surgery to excise the cyst, which led to the rapid resolution of the nerve deficit shown by clinical and electromyographical tests. A meticulous anamnesis and accurate objective examination, followed by specific tests (radiographs, sonography, and possibly CT scan) generally enable a correct diagnosis to be made. If diagnosis and therapy are carried out correctly, and without delay, symptoms quickly resolve and the nerve deficit progressively regresses. PMID- 15758859 TI - Adverse effects of GH self administration on peripheral nerve. A case report. AB - To our knowledge there is no evidence of multientrapment neuropathy related to growth hormone (GH) administration. We observed the case of a 45-year-old bodybuilder complaining of paresthesias and dysesthesias of the hands and feet after self-administration of high-dose of recombinant GH therapy. The patient complained of transitory symptoms after each cycle of GH therapy. The electrodiagnosis showed: 1) moderate right carpal tunnel syndrome (CTS) and minimal left CTS; 2) mild right ulnar entrapment at elbow; 3) a reduction of motor nerve conduction velocity of the left ulnar nerve across elbow; 4) a reduction of the amplitude of the sensory nerve action potential of the left ulnar nerve in digit-wrist segment; 5) the absence of the ulnar F wave bilaterally registering from abductor digiti minimus. Our data suggest a multiple entrapment condition of the upper limbs; furthermore, based on the history (paresthesia at feet), we can hypothesize a mild involvement of distal sensory fibres in the legs (neurophysiologically negative). The anecdotal history and neurophysiological findings of this case suggest an increased awareness to the adverse effects of GH on peripheral nerve. PMID- 15758860 TI - Acute effects of short duration maximal endurance exercise on lipid, phospholipid and lipoprotein levels. AB - AIM: The acute effects of maximal endurance exercise (15 min) on lipid and lipoprotein levels were examined, in order to determine whether the level of response produced could be affected by maximal exercise intensity (incremental stress test). METHODS: Participants in this cross sectional study were male athletes (n=78) of national level: basketball (n=10), swimming (n=9), long distance (LD) running (n=23) and wrestling (n=35); also a group of non-athlete volunteers as controls (n=19). Athletes had trained an average of 3 h/day and 5 years. The ergometric test used for the determination of maximal oxygen uptake on a treadmill was based on a stepwise stress protocol. RESULTS: Immediately after a maximal effort all groups (controls included) showed significant increases in total cholesterol (TC), triglycerides (TG), phospholipids (PL) and high-density cholesterol (HDL-C), from rest values. Low-density cholesterol (LDL-C) increased significantly in basketball, LD running and wrestling, while it did not change significantly in swimming and controls, after maximal effort. Hemoconcentration was of the order of +12.5%. CONCLUSIONS: The level of these increases might be directly related to the intensity and duration of exercise performed. Acute endurance exercise (100% VO(2max)) may induce acute modification of the above lipid parameters potentiating the systematic chronic exercise effects already present. PMID- 15758861 TI - Poor responders in assisted reproduction cycles. AB - Poor responders represent a challenging group of infertility patients who fail to respond to controlled ovarian hyperstimulation during the course of assisted reproduction treatment. Although poor response has been investigated since the eighties, many aspects of this condition are still controversial and no consensus has been reached on the management of these patients. The existing data cannot be easily compared due to the heterogeneity of the research design and methodology and the number of prospective randomized trials of sufficient sample size is limited. In addition, more research in the identification and understanding of underlying factors is needed together with a better understanding of the patho physiological basis of ovarian ageing and of the progressive reduction in ovarian reserve. This paper provides an overview on the identification, prediction and management of poor responders undergoing medicated assisted reproduction treatment. PMID- 15758862 TI - Avoidance of multiple pregnancy by use of single embryo transfer. AB - The benefits of single embryo transfer (SET) for both mother and child are evident. Already twin pregnancies constitute a relatively serious problem exemplified by the incidence of cerebral palsy, which will increase significantly as well as a risk for premature birth. Selective embryo reduction in countries where this is allowed may be one way to acutely solve the situation. In the beginning the use of natural cycle in vitro fertilization (IVF) avoided the problem, but with the introduction of controlled ovarian hyperstimulation predominantly by use of gonadotrophins in the early 1980ies the temptation to replace more than 1 embryo at a time became too strong. SET with maintenance of acceptable pregnancy rates can only be achieved if tools (improved morphological criteria, biomarkers, preimplantation genetic screening) to select the most viable/normal embryos are at hand together with improved cryopreservation procedures. In reports from Finland and Belgium already 5 years ago, elective single embryo transfer (eSET) was shown to reach almost the same success rates as double embryo transfer (DET) in selective patient groups (age under 37, good quality embryos). The indications for eSET have increased during the last years. In Finland the initiative came from the IVF clinics while in Sweden a legislative process (in act from January 2003) resulted in the recommendation that eSET should be used in the vast majority of the IVF cycles. In both these Nordic countries around 60% of the transfers are today eSET and the multiple pregnancy rate below 10% with no triplets. From an economical point of view, it is of course evident that multiple pregnancies with the numerous potential complications should be avoided altogether. Countries where IVF is included in the government health insurance will thus most likely show the way towards an even more successful, safer and cheaper treatment of infertile couples in the future. PMID- 15758863 TI - Reproductive surgery. AB - The aim of this review is to try and answer the following questions. What is the importance of reproductive surgery? What does the field encompass? What are the deleterious effects of postoperative adhesions in reproductive age women? Is there a relationship between the mode of surgical access, laparoscopy versus laparotomy, and the degree of postoperative adhesions? What is microsurgery and what is its place in reproductive surgery? Does training in microsurgery provide benefits to the gynecologist and his/her patients? At present, when assisted reproductive techniques have recorded enormous progress and the technology has been industrialized, what is the place of reconstructive tubal surgery in the treatment of tubal infertility? PMID- 15758864 TI - Use of GNRH antagonists in reproductive medicine. AB - Gonadotrophin-releasing hormone (GnRH) plays a key role in the secretion of gonadotrophins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which regulate steroidogenesis and folliculogenesis. Two GnRH antagonists, Cetrorelix and Ganirelix, deprived of histaminergic side-effects, have been introduced into ovarian stimulation protocols to prevent premature LH surges and proved their safety in clinical trials. At present, most of the published studies have not found significant differences in follicular recruitment, oocyte quality, and so on, except for a decrease in pregnancy and implantation rates in in vitro fertilization and embryo transfer (IVF-ET) cycles when the GnRH antagonist rather than the agonist was used. This decrease in pregnancy rates was in relation with a necessary learning curve of the physicians. Another possibility is the impact of the GnRH antagonist on endometrium through its GnRH receptor; this effect was cancelled after cryopreserved embryo transfers because the pregnancy rates were similar between GnRH antagonist and agonist in this case. GnRH antagonists were also interesting in poor responders and polycystic ovarian syndrome, where the agonists have not permitted to obtain the better results in IVF-ET cycles. Similarly, the GnRH antagonists could prevent the LH surge in the intrauterine insemination cycles. PMID- 15758865 TI - The diagnosis and treatment of stress-induced anovulation. AB - Behaviors that activate the hypothalamic-pituitary-adrenal (HPA) axis or suppress the hypothalamic-pituitary-thyroidal (HPT) axis can disrupt the hypothalamic pituitary-gonadal (HPG) axis in women and men. Individuals with functional hypothalamic hypogonadism typically engage in a combination of behaviors that serve as psychogenic stressors and present metabolic challenges. Complete recovery of gonadal function depends upon restoration of the HPA and HPT axes. Hormone replacement strategies have limited benefit because they do not promote recovery from these allostatic endocrine adjustments in the HPA and HPT axes. Indeed, the rationale for the use of sex steroid replacement is based on the erroneous assumption that functional forms of hypothalamic hypogonadism represent only an alteration in the hypothalamic-pituitary-ovarian (HPO) axis. Further, use of sex hormones masks deficits that accrue from altered HPA and HPT function. Long-term deleterious consequences of stress-induced anovulation may include an increased risk of cardiovascular disease, osteoporosis, depression, other psychiatric conditions, and dementia. Although fertility can be restored with exogenous administration of gonadotropins or pulsatile GnRH, fertility management alone will not permit recovery of the HPA and HPT axes. Failure to reverse the hormonal milieu induced by stress may increase the likelihood of poor obstetrical, fetal, or neonatal outcomes. In contrast, behavioral and psychological interventions that address problematic behaviors and attitudes have the potential to permit resumption of ovarian function along with recovery of the HPT and HPA axes. Full endocrine recovery offers better individual, maternal, and child health. PMID- 15758866 TI - Diagnosis and treatment of endometriosis. A review. AB - The correct approach for endometriosis management is still unclear. This review explores recent data concerning diagnosis and treatment of endometriosis, trying to define guidelines for the most appropriate diagnostic approach and therapeutic regimen. At present, laparoscopy is still considered the gold standard in endometriosis diagnosis. The risks and the diagnostic limitations of laparoscopy and the inaccuracy of clinical examination justify the considerable efforts made to improve the diagnosis with imaging techniques. The therapeutic approach is still far from being defined as causal and focuses on management of clinical symptoms of the disease rather than on the disease itself. A first-line medical therapy should be tried in patients with pelvic pain not asking for a pregnancy. Surgical treatment is considered the best treatment for women with pain and or pelvic mass who wish to become pregnant in a short time. For infertile patients, medical therapy has a limited role. The 2 treatment options include surgery or in vitro fertilization (IVF). According to our results, it seems that correct management of infertile women with endometriosis is a combination of surgery and IVF in women who did not obtain post-surgery pregnancy spontaneously. PMID- 15758867 TI - Metabolic alterations and cardiovascular risk factors in the polycystic ovary syndrome. AB - Available literature indicates that polycystic ovary syndrome (PCOS) represents a condition with a high prevalence of the metabolic syndrome, a greater and precocious appearance of glucose intolerance states, including type 2 diabetes mellitus, and a potentially greater risk for cardiovascular diseases. With respect to metabolic disorders, however, there is also emerging evidence that they are more prevalent in PCOS rather than in the general population, and that there are regional differences in the world, depending on lifestyle and other environmental factors. This evidence should be taken into consideration in future strategies focusing on prevention of metabolic and cardiovascular disorders in PCOS, not only on treatment of infertility and signs of androgen excess. PMID- 15758868 TI - The impact of altered insulin-like growth factor-I secretion on the neuroendocrine and testicular functions. AB - There is unequivocal evidence that the biosynthesis and secretion of gonadotropins by the pituitary gland is controlled by the hypothalamic GnRH and the function of the testis is mainly regulated by FSH and LH. However, a number of investigations have suggested the role of other hormones/factors, including insulin-like growth factor-I (IGF-I) in the control of pituitary and gonadal functions. The role of growth hormone (GH) and IGF-I is poorly investigated in humans. In animals with altered IGF-I secretion, the gonadotropin and androgen secretions are affected. Similarly, there is evidence that fertility, the onset of puberty and sexual maturation are affected in some patients with Laron syndrome and in acromegaly. In this minireview, we have presented some data obtained in humans and also included results from several experimental models with altered GH/IGF-I secretion, in the hope that the results from animals will possibly help in understanding the important role of IGF-I in the control of neuroendocrine-testicular function in humans. PMID- 15758869 TI - Fertility preservation in oncology. AB - Survival rates of childhood and pre-pubertal female cancer patients are constantly increasing. However the lifesaving treatments carry a significant risk for infertility. Chemotherapy and radiotherapy might induce oocyte and follicular loss, infertility and premature ovarian failure. In order to preserve the fertility potential, several options are currently available, many of those should be considered as experimental. Ovarian transposition out of the radiation field may considerably reduce the radiation dose and should be considered for patients younger than 40 years of age. The benefits of GnRH analog are not clear yet and apoptosis inhibiting agents are not available. Embryo cryopreservation is a well established technique and should be offered to patients with spouses; when the patient does not have a male partner, oocyte cryopreservation or vitrification can be performed. When the cancer treatment cannot be delayed for ovarian stimulation or the tumor is hormone sensitive then collection of immature oocytes from unstimulated ovaries is particularly useful. The oocytes are matured in-vitro and either fertilized and cryopreserved as embryos or vitrified as mature oocytes. Ovarian tissue cryopreservation has the potential of preserving thousands of primordial follicles. The thawed ovarian tissue can be autotransplented orthotopically or heterotopically. Until now, only one human live birth has been reported and critical issues like the potential risk of transplanting malignant cells and the survival of the grafts have to be addressed. The strategy for preservation of fertility prior to cancer treatment should be tailored according to the patients age, presence of a partner, type of malignant disease, therapeutic agent, and time interval available. The patient should obviously be informed that some of the methods are still experimental. PMID- 15758870 TI - Carotid artery stenting for the high surgical risk patients. AB - Obstructive atherosclerotic carotid bifurcation disease is responsible for approximately 30% of stroke cases in the United States. Any successful treatment of this disease should demonstrate significant and durable reduction in the related risk for stroke without compromising the patient safety. The indications for CAS are evolving as the techniques, the stents and catheter technology advance. There is an ongoing need to improve the equipment suitable for carotid stenting and a need to enhance the safety of the procedure. Often, when a technical problem is encountered, it is attributed to the inadequacy of the devices currently available. As the technology improves, particularly with the application of the anti-embolization devices, the indications and contraindication may need to be revised. PMID- 15758871 TI - Endovascular stent graft selection for the treatment of abdominal aortic aneurysms. AB - Minimally invasive endovascular techniques for the treatment of abdominal aortic aneurysms (AAA) have significantly reduced the morbidity of these procedures as compared with standard surgical repair. In addition, patients with extensive comorbid medical illnesses in whom standard operative repair is contra-indicated, may be successfully treated using endovascular means. A variety of endovascular stent grafts are currently being used clinically for endovascular AAA repair. The characteristics of these stent grafts vary significantly. In selecting the specific stent graft to be used for endovascular AAA repair, these specific characteristics must be considered particularly with regard to the individual patient's anatomic and physiologic characteristics. The United States Food and Drug Administration (FDA) has approved 4 endovascular stent grafts for the treatment of AAA: the Medtronic AneuRx, the W. L. Gore Excluder, the Cook Zenith and the Guidant Ancure. The Zenith graft received approval for marketing in 2003 and has gained relatively wide usage, in part due to its ability to treat aneurysms with relatively large diameter implantations zones. Also in 2003 the Guidant Corporation withdrew the Ancure graft from marketing and distribution. Enrollment has also been recently completed for the phase II FDA trials of the Medtronic Talent graft and the Cordis Fortron graft. These devices have been approved for use in the European Union and are awaiting FDA panel meeting in the United States. Comparative analysis of several of the available stent grafts has been performed. Results have varied with regard to the need for secondary interventions, aneurysm sac size reduction and the occurrence of continued perfusion of the aneurysm sac. Several significant advances have also occurred recently in stent graft research and development. Of particular significance has been the initiation of the phase I trial of the Trivascular Enovus graft. Deployment of the Trivascular graft may be accomplished through a delivery system that is considerably reduced in profile. The potential for percutaneous application of the graft may be available in the future. The indications for use of endovascular grafts as compared to standard open surgery have not yet been fully defined. Endovascular stent grafts in current use have limitations and their use must be tempered accordingly, until their long-term effectiveness is more completely evaluated. This article describes the general principles of use for endovascular devices for the repair of AAAs. It details the features and results for the devices in current use and highlights the factors that influence the selection of specific stent graft types. PMID- 15758872 TI - Differentiated treatment of aneurysms of the extracranial carotid artery. AB - AIM: Aim of the study is to illustrate current therapeutic options for aneurysms of the extracranial carotid artery. METHODS: From 1987 until 2003 14 patients (average age: 60 years) with an aneurysm of the extracranial internal carotid artery were treated. The aneurysm diameter ranged from 5 mm to 60 mm. Thirteen patients underwent elective surgery. One patient was operated upon because of acute severe cervical bleeding. RESULTS: Most patients (10/14) presented with an asymptomatic pulsatile cervical mass. The aneurysm was diagnosed 3 times during carotid angiography. The underlying etiology was atherosclerosis in 10 patients. In 3 cases the aneurysm was considered mycotic. Ten aneurysms were localized at the carotid bifurcation whereas 2 each were found in the retrostyloideal region and at the base of the skull. Vascular continuity could be restored in 10 patients while in 3 the carotid artery had to be occluded (twice surgically and once radiologically). The final patient underwent stent insertion for post traumatic distal aneurysm. In 5 patients perioperative neurological complications were noted which persisted in a single patient (dysphagia due to a lesion of the hypoglossus nerve). All hemispheric complications recovered completely. During follow-up (median 48 months ranging from 3 to 103 months), 1 patient died of unrelated cause. Another patient developed a contralateral ischemic insult. CONCLUSIONS: When adjusting treatment to the various etiological conditions and localizations, aneurysms of the extracranial carotid artery can be managed with a low complication rate and satisfying long-term results. However, success mainly depends on the timely diagnosis and availability of a range of surgical and radiological therapeutical options. PMID- 15758873 TI - Hemobahn stent-grafts in the treatment of femoropopliteal occlusive disease. AB - AIM: The aim of this study was to determine our results and indications for the Hemobahn stent-graft in femoropopliteal occlusive disease. METHODS: Since 1999, 38 patients with femoropopliteal occlusive disease in 40 legs have been treated by endovascular dilation or recanalisation and additional endografting. All have been followed up prospectively at 6 month intervals by clinical examination, Doppler and duplex. RESULTS: In 40 legs, 15 high grade stenoses were dilated and 25 occlusions needed first recanalization. Additional Hemobahn stent-graft deployment was successful in all cases (total grafts n=60). The median length of the stented segment was 15 cm. Immediate complications during the procedure were stent-graft thrombosis (n=1) and distal embolization (n=3), instantly treated with thrombolysis and thrombus aspiration. The median ankle-brachial index in rest increased from 0.5 preoperatively to 1 postoperatively. With a median follow up time of 28.2 months (range from 1 to 48 months), there were 4 high grade stenoses and 13 stent-graft occlusions (1 year primary patency rate of 66%). All 4 stenoses were treated with balloon dilation (1 year assisted primary patency rate of 76%) and 4 of the occluded grafts were recovered by thrombectomy (n=1) or thrombolysis (n=3), resulting in a 1 year secondary patency rate of 87%. CONCLUSIONS: The Hemobahn stent-graft can be used in the treatment of femoropopliteal occlusive disease with a low complication rate and acceptable patency rates in the short-term. We had the best results when there was no recanalization needed before. PMID- 15758874 TI - Cardiac risk assessment of asymptomatic patients by stress echocardiography before infrarenal aortic aneurysm surgery. AB - AIM: Aggressive cardiac assessment before aortic abdominal aneurysm (AAA) surgery is indicated for patients with symptomatic coronary artery disease (CAD). Assessment of intermediate and moderate risk patients is still under debate. The purpose of the study was to prospectively evaluate the effectiveness of stress echocardiography (SE) in the detection of CAD in patients undergoing AAA surgery who have no symptoms and/or signs of CAD, but who have risk factors for it. METHODS: Patients with 1 or more risk factors for CAD underwent SE. All patients with positive SE underwent coronary arteriography, and, when indicated, treatment. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for SE by comparing results to coronary arteriography. Moreover, major perioperative cardiac events were recorded. RESULTS: Ninety-one patients with AAA and risk factors for CAD were studied. SE was positive in 9 cases, including 7 presenting critical CAD on the basis of coronary arteriography. One major cardiac event (1.1%), a nonfatal myocardial infarction, occurred in 1 patient with positive SE and non-critical, single-vessel CAD. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of SE proved to be 100%, 98%, 78%, 100%, and 92%, respectively. CONCLUSIONS: Positive SE should be considered a valid method for testing high-risk patients for CAD. The low rate of major cardiac events in this series suggests that cardiac assessment by SE and selective coronary arteriography prior to AAA surgery is effective in asymptomatic patients with one or more risk factors. PMID- 15758876 TI - Pseudoaneurysm of the popliteal artery. Surgical management of 3 cases. AB - Pseudoaneurysm of the popliteal artery is much less common than true aneurysm in this location. We report 3 cases of popliteal artery pseudoaneurysms, each of differing etiopathology, one presenting 3 months after trauma, one following aortic valve replacement for aortic regurgitation with suspected endocarditis, with no organism detected, and the 3rd of infective etiology. The clinical presentation and surgical management are discussed with a review of the literature. PMID- 15758875 TI - MRSA in lower limb amputation and the role of antibiotic prophylaxis. AB - AIM: Methicillin Resistant Staphylococcus Aureus (MRSA) colonisation is reported in 3-20% of vascular patients. Many develop infective complications. MRSA is associated with poor prognosis. Aim of the study is to assess MRSA in lower limb amputation and efficacy of antibiotic prophylaxis. METHODS: Prospective study of lower limb amputation. MRSA screen and wound swabs were taken at operation. Antibiotic prophylaxis included teicoplanin (400 mg) 1 dose at operation. RESULTS: Twenty-five patients underwent 33 primary amputations. At operation 15 legs (45%) were colonised with MRSA and 18 legs (58%) had active wound infection; MRSA (4) and other (14). Following surgery 3 patients died. Twenty-two legs (76%) had primary healing. Infection developed in 7 stumps (24%), MRSA (5) and Pseudomonas (2). Stump infection increased time to wound healing (p<0.0001). MRSA stump infection increased revision amputation (p=0.009) and duration of hospital stay (p<0.0074). MRSA wound infection at operation increased the risk of MRSA stump infection (p=0.007). Non-MRSA wound infection at operation was not associated with a worse outcome. No patient colonised with MRSA at operation developed postoperative MRSA stump infection. CONCLUSIONS: MRSA is more prevalent that previously reported. MRSA infection has a poor prognosis. Prophylaxis may be effective for patients colonised with MRSA. PMID- 15758877 TI - Venous reconstruction of the inferior vena cava bifurcation for retroperitoneal rhabdomyosarcoma in a child. Case report. AB - Retroperitoneal tumors, as well as traumatic lesions and occlusions of the bifurcation of the inferior vena cava (IVC) and the common iliac veins may require venous vascular reconstruction. We present a method for inferior vena caval bifurcation reconstruction which employs the advantages of a large size straight expanded polytetrafluorethylene (ePTFE) graft in this position after a new IVC bifurcation has been created by uniting the stumps of both common iliac veins, and construction of an arteriovenous (A-V) fistula with controlled flow to decrease the rethrombosis rate of the graft, and still not cause heart failure. This method was used in a case of recurrent rhabdomyosarcoma in an 8 year old child encroaching upon the bifurcation of the IVC as well as on both common iliac veins. The situation was managed by radical resection of the tumor and by creating a new caval bifurcation of the common iliac veins, followed by interposition of a straight ePTFE graft. An A-V fistula was created between the left femoral vein and left femoral artery using the left greater saphenous vein in controlled fashion. The arterial defect of the right common iliac artery was reconstructed by interposition of a PTFE graft end-to-end. PMID- 15758878 TI - Anterolateral right thoracotomy for mitral valve procedure after previous coronary artery bypass grafting with functioning internal mammary artery grafts. AB - AIM: Mitral valve procedure after previous coronary artery bypass grafting (CABG) with functioning internal mammary artery (IMA) grafts has high risk. Especially, internal mammary artery grafts injury may be fatal. The anterolateral right thoracotomy affords easy access to the right atrium with minimal dissection, and minimizes the risk of injury to the IMA grafts. We reviewed our operative technique and outcome after mitral valve procedure after previous CABG with functioning IMA grafts. METHODS: Thirteen patients (11 male and 2 female, mean age of 67.7+/-8.5 years, range 54 to 80 years) underwent mitral valve replacement after previous CABG with functioning IMA grafts from march 1993 to september 2002. The mean interval between the previous CABG and the mitral valve procedure was 3.8 years (range 9 months to 8 years). Four patients had simultaneous mitral valve procedures at initial CABG (2 repairs and 2 replacements). The operation has performed through the anterolateral right thoracotomy, under ventricular fibrillation with moderate hypothermia and without cardioplesia. RESULTS: Mitral valve repair was performed in 3 patients, mitral valve replacement in 10 patients. The mean coronary bypass time was 69.1+/-16.2 min (range 45 to 98 min). The operation time was 159.3+/-29.4 min (range 120 to 219 min). Intensive care unit stay days was 1.9+/-1.6 days (range 1 to 5 days). Peak CK/CK-MB values were 555.1+/-290.4 IU/16.6+/-10.7 IU (range 176 to 924 IU/7 to 44 IU). Peak troponin I value was 9.5+/-5.2 pg/mL (range 4 to 17.8 pg/mL). There was no IMA injury and no early death. Other complications were newly arrhythmia in 3 patients, renal insufficiency in 1 patient, reoperation for bleeding in 1 patient. CONCLUSIONS: Anterolateral right thoracotomy approach, ventricular fibrillation with moderate hypothermia without cardioplesia were a safe and good method for mitral valve operation after previous CABG with functioning IMA graft. PMID- 15758879 TI - The use of preoperative intra-aortic balloon pump in open heart surgery. AB - AIM: Intra-aortic balloon pump (IABP) therapy before open heart surgery has been suggested for ''high risk'' patients. METHODS: Records from patients undergoing open heart surgery at our institution between June 1999 and February 2002 were reviewed. Indication for IABP insertion was severely impaired left ventricular function, acute myocardial infarction (MI) or unstable angina. RESULTS: Fifty five patients were included in the study: 41 male, 14 female, age 64+/-9 years. Fifty-one (92.7%) required coronary artery bypass brafting (CABG) alone or as a combined procedure, 2 (3.6%) required mitral valve surgery, and 2 (3.6%) needed more complex cardiac procedures. Thirty-two patients (58%) underwent emergency cardiac surgery and 11 patients (20%) suffered from acute preoperative MI. The overall 30 days mortality was 9%. Mean intensive care unit (ICU) stay was 6+/-8 days. Four patients (7.2%) developed postoperative renal failure requiring temporary hemodialysis. Three patients (5.4%) developed IABP related peripheral vascular complications. CONCLUSIONS: Perioperative morbidity and mortality is increased despite preoperative IABP, particularly in patients with acute MI. In contrast to studies not using this approach, preoperative IABP reduces morbidity and mortality of high risk patients. IABP related complications are low. Our data suggest that high risk patients profit from preoperative IABP therapy, however, prospective studies are needed to confirm these findings. PMID- 15758880 TI - Initial, continuous and intermittent bolus cardioplegia administration: efficacy of potassium-chloride and magnesium-sulfate as minimal additives for minimally diluted blood cardioplegia. AB - AIM: The most effective delivery of blood cardioplegia (BCP) remains controversial, and a combination of initial continuous and intermittent bolus BCP seems to compensate each demerit. However, a large amount of crystalloid solution is infused into the myocardium in this method, which may nullify the advantages of BCP. We examined the hypothesis that minimally-diluted BCP resolves this issue and provides superior myocardial protective effects. METHODS: Seventy patients undergoing elective coronary revascularization between 1997-2001 (M:F=55:15, mean age 67.6+/-7.5 years) were randomly allocated into one of 2 groups: Group C (n=35) was given the standard 4:1-diluted blood-crystalloid BCP, and Group M (n=35) was given minimally-diluted BCP supplemented with potassium-chloride and magnesium-sulfate. The BCP temperature was maintained at 30 degrees C. Cardioplegic arrest was induced with 2 minutes of initial antegrade BCP infusion, followed by continuous retrograde BCP infusion. Intermittent antegrade BCP was infused every 30 minutes for 2 minutes. RESULTS: The time required for achieving cardioplegic arrest was significantly shorter in Group M (47.5+/-16.3 vs 62.5+/ 17.6 s, p<0.0001) and the number of patients showing spontaneous heart-beat recovery after aortic unclamping was significantly larger in Group M (28 vs 15, p=0.0029). The number of patients suffering from atrial fibrillation during the postoperative period was significantly smaller in Group M (3 vs 11, p=0.034). The total amount of crystalloid solution infused as cardioplegia was significantly smaller in Group M (62.8+/-22.3 vs 733.6+/-382.6 mL, p<0.0001). Postoperative maximum dopamine dose (3.57+/-2.46 vs 5.44+/-2.23 mg/kg/min, p=0.0014) and peak creatine kinase-MB (19.5+/-8.5 vs 25.8+/-11.9 IU/L, p=0.0128) were significantly lower in Group M. The number of patients showing paradoxical movement of the ventricular septum by early postoperative echocardiography was significantly smaller in Group M (9 vs 24, p<0.0007). CONCLUSIONS: These results demonstrate that initial continuous and intermittent bolus administration of minimally diluted BCP supplemented with potassium and magnesium can be a simple, reliable and effective technique of intraoperative myocardial protection. PMID- 15758881 TI - Intraoperative treatment of chronic atrial fibrillation with ultrasound. AB - AIM: The purpose of this retrospective study is to evaluate the efficacy of intraoperative atrial compartmentalization utilizing ultrasound (US) energy in selected patients with chronic atrial fibrillation (CAF). METHODS: From March 1999 to June 2000, 27 patients with ages ranging from 15 to 69 (mean 44.62+/ 15.61 years), 15 of whom were female, underwent intraoperative atrial compartmentalization with US. Mitral valve replacement was performed in 22 patients, mitral valve repair in 4 and 1 patient with paroxysmal atrial fibrillation (AF) underwent isolated atrial compartmentalization. The compartmentalization procedure was started using an ultrasonic device (UltraCision) at the level of the right atrium on cardiopulmonary bypass (CPB) and with beating heart, and then, after the mitral valve procedure, the ablation lines were traced in an inverted U-shape, involving the 4 pulmonary veins. All patients were given verapamil or amiodarone for electrical atrial remodeling for 6 months. RESULTS: Time of surgery was 166 min on average, that is approximately 79 min of CPB, 59 min of aortic crossclamping, 12.5 min for right atrial lesion lines and 14 min for the left ones. After weaning CPB, all the patients maintained adequate cardiac output. Immediate reversion to sinus rhythm was achieved in 24 patients (88.8%). A small intraoperative left atrial wall rupture due to the ablation technique was easily repaired by a direct continuous suture. Another complication observed in 1 patient was a sternal instability, which was fixed successfully. Two patients died because of postoperative respiratory failure due to bilateral pneumonia in 1 case and low cardiac output, complicated by septicemia. The pharmacological anti-arrhythmic treatment was stopped 6 months after surgery. During 29 months of follow-up, 20 patients (74%) were still in sinus rhythm. CONCLUSIONS: The use of US during mitral valve operation in patients with CAF allowed for easy and rapid reestablishment of the sinus rhythm. This technique modifies and makes the Maze procedure easier, decreases surgical time and the incidence of postoperative complications. PMID- 15758882 TI - Redo aortic valve replacement with ''porcelain'' aorta in an aortitis patient. A case report. AB - We report a successful reoperation of aortic valve replacement with the severely calcified, ''porcelain'' aorta due to Takayasu's disease. The patient was a 46 year-old female diagnosed with aortic prosthetic valve failure. Takayasu's aortitis had been noted at 17 years old and the patient underwent aortic valve replacement for aortic regurgitation at age 32. Chest CT scan revealed severe circumferential calcification of the whole thoracic aorta. Echocardiography revealed 100 mmHg of pressure gradient across the aortic prosthetic valve with pannus formation. Replacement of the aortic prosthetic valve was performed using the selective cerebral perfusion method under deep hypothermia. The postoperative course was uneventful without any neurological disorder. Selective cerebral perfusion was a useful method to avoid cross clamping in severely calcified ascending aortic. PMID- 15758883 TI - Off-pump coronary artery bypass in a patient with the antiphospholipid syndrome. AB - Patients with antiphospholipid syndrome (APS) are prone to excessive postoperative morbidity and mortality after cardiovascular surgery because of its thromboembolic derangements. We present a case of coronary artery bypass grafting (CABG) in a patient with primary APS. He suffered from repetitive coronary occlusion after percutaneous transluminal coronary angioplasty (PTCA). Since his lupus anticoagulant level was found to be 217 s (normal, <50 s), he was diagnosed as the primary APS. He received steroid pulsation therapy with 1000 mg of prednisolone, double-filtration plasmapheresis (DFPP) and 50 mg of cyclophosphamide to attenuate the antibody activity. Four months after the last PTCA, he experienced chest pain and approximately 90% of stenosis in the left anterior descending (LAD) lesion was apparent, although the titer for the lupus anticoagulant was reduced to the normal range. He had drug allergy to ticlopidine hydrochloride and aspirin. Taken together, his disease was found to be resistant to these medical treatments, and surgical treatment was considered. Since cardiopulmonary bypass is known to exaggerate its coagulatory and fibrinolytic complications, off-pump CABG (OPCAB) was feasible in this case. The left internal thoracic artery (ITA) was anastomosed to the LAD using the off-pump technique. The procedure was successful, and the postoperative course for 3 years has been satisfactory without any cardiovascular complaints. PMID- 15758884 TI - Concomitant endovascular treatment of thoracic and abdominal aortic aneurysms. PMID- 15758885 TI - Endoleak after open abdominal aortic aneurysm repair: new open repair following unsuccessful endoluminal treatment. PMID- 15758886 TI - Primary supraceliac aortic pseudoaneurysm due to a septic aortic rupture. PMID- 15758887 TI - The Corx System for CABG of thoracic and abdominal aortic aneurysms. PMID- 15758888 TI - Management of the failing heart during console failure. PMID- 15758889 TI - A total circumferential tear of Stanford type a dissection with a prolapsing flap into the left ventricular outflow tract: a potential cause of exaggerating aortic regurgitation. PMID- 15758890 TI - Mediastinitis due to Mycobacterium tuberculosis after a redo open heart surgery. PMID- 15758891 TI - Lymphangioleiomyomatosis presenting as a refractory chylothorax. PMID- 15758892 TI - Successful management of idiopathic fibrosing mediastinitis with superior vena cava thrombosis. PMID- 15758893 TI - Inadvertent use of Bicillin C-R to treat syphilis infection--Los Angeles, California, 1999-2004. AB - In March 2004, the Los Angeles County Department of Health Services (LACDHS) was notified that a large nonprofit clinic serving the gay and lesbian community in Los Angeles used a nonrecommended preparation of penicillin to treat syphilis patients during January 1999-March 2004. The clinic had inadvertently used Bicillin C-R, a mixture of 1.2 million units (MU) benzathine penicillin G (BPG) and 1.2 MU procaine penicillin G, rather than Bicillin L-A, a preparation that contains the 2.4 MU BPG per dose recommended by CDC. Bicillin L-A is recommended for treating syphilis and upper respiratory tract infections caused by susceptible streptococci. Bicillin C-R is indicated for streptococcal infections of the skin and respiratory tract; however, its efficacy in treating syphilis is unknown. The inadvertent use of Bicillin C-R, which contains only half the recommended dose of BPG for syphilis, was discovered after a patient treated for syphilis read the product insert, which stated that the medication was not indicated for treatment of syphilis. Review of clinic pharmacy records revealed that it received a shipment of Bicillin C-R in lieu of an unfilled order for Bicillin L-A in late 1998 and that the pharmacy subsequently ordered Bicillin C-R until March 2004. The clinic used Bicillin C-R as its exclusive formulation of injectable penicillin during January 1999-March 2004. This report summarizes the investigation of the misuse of Bicillin C-R at the Los Angeles clinic, which represents the largest occurrence of inadvertent treatment with Bicillin C-R to date. The investigation led to discussions among CDC, the Food and Drug Administration (FDA), and King Pharmaceuticals, Inc. (Bristol, Tennessee), whose Monarch Pharmaceuticals subsidiary markets Bicillin products. As a result, King Pharmaceuticals agreed to institute packaging and labeling changes to Bicillin products to prevent inadvertent treatment of syphilis with Bicillin C-R. PMID- 15758894 TI - Transmission of hepatitis B virus among persons undergoing blood glucose monitoring in long-term-care facilities--Mississippi, North Carolina, and Los Angeles County, California, 2003-2004. AB - Regular monitoring of blood glucose levels is an important component of routine diabetes care. Capillary blood is typically sampled with the use of a fingerstick device and tested with a portable glucometer. Because of outbreaks of hepatitis B virus (HBV) infections associated with glucose monitoring, CDC and the Food and Drug Administration (FDA) have recommended since 1990 that fingerstick devices be restricted to individual use. This report describes three recent outbreaks of HBV infection among residents in long-term-care (LTC) facilities that were attributed to shared devices and other breaks in infection-control practices related to blood glucose monitoring. Findings from these investigations and previous reports suggest that recommendations concerning standard precautions and the reuse of fingerstick devices have not been adhered to or enforced consistently in LTC settings. The findings underscore the need for education, training, adherence to standard precautions, and specific infection-control recommendations targeting diabetes-care procedures in LTC settings. PMID- 15758895 TI - Salmonellosis associated with pet turtles--Wisconsin and Wyoming, 2004. AB - Salmonellosis associated with small pet turtles in the United States was a major public health concern in the 1970s. In 1975, the Food and Drug Administration (FDA) banned commercial distribution of small turtles (i.e., those with a carapace of <4 inches). The FDA ban prevents an estimated 100,000 cases of salmonellosis among children each year. However, a recent resurgence in the sale of small turtles has generated concern. In Wisconsin and Wyoming, at least six human cases of salmonellosis have been linked to such turtles. This report describes the investigation into those cases. The findings underscore the need for health and environmental officials to prevent illegal distribution of small turtles and consider patient contact with turtles when investigating salmonellosis cases. PMID- 15758896 TI - Lead poisoning associated with use of litargirio--Rhode Island, 2003. AB - Lead can damage the neurologic, hematologic, and renal systems. Deteriorated leaded paint in older housing remains the most common source of lead exposure for children in the United States; however, other lead sources increasingly are recognized, particularly among certain racial/ethnic populations. In 2003, the Rhode Island Department of Health (RIDOH) recognized litargirio (also known as litharge or lead monoxide), a yellow or peach-colored powder used as an antiperspirant/deodorant and a folk remedy in the Hispanic community, as a potential source of lead exposure for Hispanic children. This report summarizes a case investigation of elevated blood lead levels (BLLs > or =10 microg/dL) associated with litargirio use among two siblings in Rhode Island, the public health action taken, and a survey of parents/guardians in three pediatric clinics in Providence, Rhode Island, to assess litargirio use. Findings underscore the importance of follow-up of elevated BLLs and thorough investigation to identify all lead sources. PMID- 15758897 TI - Care at the front line: clinical decisions in the management of pediatric acid related disorders. PMID- 15758898 TI - Asthma and gastroesophageal reflux disease in children: exploring the relationship. AB - Asthma is a well-recognized disease and one of the most common illnesses in childhood. More recently, gastroesophageal reflux disease has increasingly been appreciated as a common daily occurrence in children and adolescents. These two diagnoses often present in tandem, with their coexistence being more frequent than would be expected for a chance occurrence. The mainstay of asthma management is the regulation and control of chronic airway hyperreactivity and inflammation. Children who do not respond to standard asthma regimens should be evaluated for other sources of their pulmonary symptoms, most notably gastroesophageal reflux. Baseline assessment of pulmonary function tests followed by an empiric trial of proton pump inhibitor therapy, using double the standard doses commonly used in acid-related disorders and administered for 3 months, is a cost-effective, noninvasive diagnostic strategy. Children who fail to exhibit pulmonary symptom improvement should be evaluated for both medication compliance and proper administration. Twenty-four-hour esophageal pH monitoring with concurrent dairy recordings of their symptoms is recommended to ascertain adequacy of acid suppression and confirm the diagnosis in those who continue to have symptoms. Children with acid-related causes of their pulmonary symptoms often require long term treatment. Studies have confirmed the efficacy, safety, and tolerability of proton pump inhibitors in the treatment of children and adolescents. Surgery should be reserved for those with severe disease and those who are unable to comply with pharmacologic treatment. PMID- 15758899 TI - Helicobacter pylori infection: detection, investigation, and management. AB - Helicobacter pylori infection causes gastritis and peptic ulcers and is associated with the development of gastric cancer. Approximately 50% of the world population is infected with H pylori , with the highest prevalence rates in developing countries. In the vast majority of individuals, infection is acquired during childhood with those of low socioeconomic means and having infected family members being at highest risk for early childhood acquisition. Definitive routes of transmission of the infection are unclear, with evidence suggesting oral-oral, gastric-oral, and fecal-oral routes. If untreated, H pylori infection is lifelong. Although clinical disease typically occurs decades after initial infection acquisition, children infected with H pylori may have gastritis, ulcers, mucosal-associated lymphoid type lymphoma, and, rarely, gastric atrophy with/without intestinal metaplasia (ie, both precursor lesions for gastric cancer). Controversy persists regarding testing for and treating H pylori , if found, in the large number of children who present with recurrent abdominal pain. Because young children (ie, younger than 5 years of age) who are treated and cured of their H pylori infection may be at risk for reinfection, the current recommendations do not recommend treatment unless an ulcer or gastric atrophy is present. However, despite the lack of clinical evidence, the trend is to more aggressively screen children for the presence of H pylori and to treat those children who are found to have the infection. H pylori infection can be eradicated by antimicrobial therapy plus a proton pump inhibitor, but no treatment regimen is 100% effective. Multiple drugs, frequent dosing, and length of treatment often contribute to poor patient compliance, and antibiotic eradication therapy is associated with increasing drug resistance. PMID- 15758900 TI - Decisions in diagnosing and managing chronic gastroesophageal reflux disease in children. AB - Gastroesophageal reflux disease (GERD) presents in different ways in children, most commonly with vomiting, or with esophageal symptoms such as regurgitation, heartburn, or dysphagia. Extraesophageal symptoms and signs also frequently occur. Less well recognized is that abdominal pain is a relatively common mode of presentation. Although abdominal pain is common in school-aged children, GERD and other acid-related disorders such as peptic ulcer disease are relatively uncommon causes of such. A careful history will usually determine whether an acid-related disorder is in the differential diagnosis of abdominal pain. Early detection and treatment of GERD in children may prevent, attenuate, or heal complications such as failure to thrive or feeding refusal as well as pulmonary, ear-nose-and-throat disorders, erosive esophagitis, and peptic stricture. In children with persistent or severe symptoms and/or complications of GERD such as erosive esophagitis, the major treatment options are pharmacologic management with acid-suppressing medication, specifically proton pump inhibitors (PPIs), or antireflux surgery. For many patients, PPI treatment offers advantages over surgery. When given in adequate doses, PPIs can safely effect relief of GERD symptoms and healing of esophagitis in children. Antireflux surgery may work well in selected patients, but it carries significant risk of morbidity, including high failure rates, even in the short term. Some postoperative studies report that more than 60% of patients are back on medical treatment with proton pump inhibitors for recurrence of GERD symptoms, and a similar percentage have new symptoms that were not present before surgery. Death is uncommon but does occur and is an unacceptable risk in an otherwise healthy, low-risk individual. Laparoscopic surgery may have some disadvantages compared with open surgery, including a higher rate of redo operations. Studies show that many children undergo surgery for unclear indications, often with few preoperative diagnostic studies. The availability of highly effective medical therapy, together with more careful selection of patients for surgery, may result in better patient outcomes, with much lower operative rates. PMID- 15758903 TI - ASGE Guideline: Guidelines for endoscopy in pregnant and lactating women. PMID- 15758904 TI - ASGE guideline: The role of endoscopy in the diagnosis and the management of cystic lesions and inflammatory fluid collections of the pancreas. PMID- 15758905 TI - Colonic biopsy practice for evaluation of diarrhea in patients with normal endoscopic findings: results from a national endoscopic database. AB - BACKGROUND: The colonic biopsy is the only reliable method for identification of microscopic colitis in patients with chronic diarrhea and normal endoscopic findings. METHODS: The Clinical Outcomes Research Initiative national endoscopic database was analyzed to determine the rate at which colonic biopsy specimens were obtained in patients undergoing colonoscopy for the evaluation of diarrhea with no visible mucosal abnormality. RESULTS: Between January 2000 and December 2003, 5565 unique adult patients underwent colonoscopy for evaluation of diarrhea without detection of any mucosal abnormality. Colonic mucosal biopsy specimens were obtained in 4410 (79.2%) of these patients. The rates at which biopsy specimens were obtained differed among the sites where colonoscopy was performed; biopsy specimens were obtained from more patients undergoing colonoscopy in university-affiliated settings (86.8%) compared with Veterans Affairs Medical Centers (VAMC) (78.5%) or community sites (78.6%) ( p < 0.001). On multivariate analysis, biopsy specimens were more likely to be obtained in younger patients (OR 0.7: 95%CI[0.6, 0.8] for age >50 years vs. <50 years), women patients (OR 1.4: 95% CI[1.2, 1.6] in community setting; OR 4.1: 95% CI[1.6, 10.5] in VAMC setting), and patients seen in university-affiliated medical centers (university center OR 2.1: 95% CI[1.5, 3.0] vs. community setting). CONCLUSIONS: Biopsy specimens are obtained in four fifths of patients with diarrhea and normal colonoscopy findings to exclude microscopic colitis. Variation in biopsy practice exists among endoscopy site types and by gender. Clear guidelines are needed for the endoscopic approach to these patients. PMID- 15758906 TI - Chronic diarrhea: To biopsy or not to biopsy. PMID- 15758907 TI - Impact of colonic cleansing on quality and diagnostic yield of colonoscopy: the European Panel of Appropriateness of Gastrointestinal Endoscopy European multicenter study. AB - BACKGROUND: The quality of colon cleansing is a major determinant of quality of colonoscopy. To our knowledge, the impact of bowel preparation on the quality of colonoscopy has not been assessed prospectively in a large multicenter study. Therefore, this study assessed the factors that determine colon-cleansing quality and the impact of cleansing quality on the technical performance and diagnostic yield of colonoscopy. METHODS: Twenty-one centers from 11 countries participated in this prospective observational study. Colon-cleansing quality was assessed on a 5-point scale and was categorized on 3 levels. The clinical indication for colonoscopy, diagnoses, and technical parameters related to colonoscopy were recorded. RESULTS: A total of 5832 patients were included in the study (48.7% men, mean age 57.6 [15.9] years). Cleansing quality was lower in elderly patients and in patients in the hospital. Procedures in poorly prepared patients were longer, more difficult, and more often incomplete. The detection of polyps of any size depended on cleansing quality: odds ratio (OR) 1.73: 95% confidence interval (CI)[1.28, 2.36] for intermediate-quality compared with low-quality preparation; and OR 1.46: 95% CI[1.11, 1.93] for high-quality compared with low-quality preparation. For polyps >10 mm in size, corresponding ORs were 1.0 for low quality cleansing, OR 1.83: 95% CI[1.11, 3.05] for intermediate-quality cleansing, and OR 1.72: 95% CI[1.11, 2.67] for high-quality cleansing. Cancers were not detected less frequently in the case of poor preparation. CONCLUSIONS: Cleansing quality critically determines quality, difficulty, speed, and completeness of colonoscopy, and is lower in hospitalized patients and patients with higher levels of comorbid conditions. The proportion of patients who undergo polypectomy increases with higher cleansing quality, whereas colon cancer detection does not seem to critically depend on the quality of bowel preparation. PMID- 15758909 TI - Quality and colonoscopy: a new imperative. PMID- 15758908 TI - Analysis of colorectal cancer occurrence during surveillance colonoscopy in the dietary Polyp Prevention Trial. AB - BACKGROUND: Interval colorectal cancer (CRC) occasionally is detected in patients who have recently undergone colonoscopy. Systematic evaluation of CRC detected after colonoscopy could identify ways to improve the quality and the outcome of colonoscopy. METHODS: This study examined cancer diagnoses in the course of the dietary Polyp Prevention Trial, a randomized study of a dietary intervention on recurrence of adenomatous polyps. An algorithm was developed to classify each cancer into one of 4 etiologies: (1) incomplete removal (cancer at the site of previous adenoma), (2) failed biopsy detection (cancer in an area of suspected neoplasia with negative biopsy specimens), (3) missed cancer (large, advanced stage cancer found at a short interval after colonoscopy), or (4) new cancer (small, early stage cancer after a longer time interval). RESULTS: Of 2079 patients, 13 had cancer detected over 5810 person years of observation (PYO) (2.2 cases/1000 PYO); 7/13 or 53.8% of patients had either a potentially "avoidable" cancer or one detectable at an earlier time interval because of incomplete removal (4/13) or missed cancer (3/13). CONCLUSIONS: Interval cancer occurs despite colonoscopy. Improved quality of colonoscopy may have reduced cancer prevalence or resulted in earlier cancer detection in over 50% of prevalent cancers in the dietary Polyp Prevention Trial. PMID- 15758910 TI - 18-Fluorodeoxyglucose positron emission tomography has limited sensitivity for colonic adenoma and early stage colon cancer. AB - BACKGROUND: 18-Fluorodeoxyglucose positron emission tomography (PET) is used clinically to detect recurrent colon cancer after surgical resection, but the sensitivity of PET for premalignant colon lesions and early stage colon cancer is not well defined. METHODS: In a prospective study, 45 patients with a total of 58 colonic neoplasms, including premalignant polyps, premalignant, flat lesions, and early stage cancers, were evaluated by PET. RESULTS: The sensitivity of PET for cancer was 62% (8/13). PET detected 100% (7/7) of cancers 2 cm or larger but only 17% (1/6) of cancers smaller than 2 cm. PET detected 23% (3/13) of flat, premalignant lesions; 70% (7/10) of protruded, premalignant lesions 3 cm or larger; 38% (3/8) of protruded, premalignant lesions between 2 and 2.9 cm; and 14% (2/14) of protruded, premalignant lesions between 1 and 1.9 cm. There was no false-positive PET reading. CONCLUSIONS: PET has limited sensitivity for flat, premalignant lesions; protruded, premalignant lesions smaller than 3 cm; and colon cancers smaller than 2 cm. PMID- 15758911 TI - Age-related changes in the pancreas identified by EUS: a prospective evaluation. AB - BACKGROUND: EUS is an important modality for the diagnosis of pancreatic disease. An understanding of normal pancreatic ductal and parenchymal variation in asymptomatic individuals is essential to improve EUS accuracy. The primary aim of this study was to determine age-related pancreatic parenchymal and ductular changes identifiable on EUS in individuals with no history or symptoms of pancreaticobiliary disease. Secondary aims were to define demographic and clinical factors associated with identifiable pancreatic parenchymal and ductular changes, and to determine the main pancreatic-duct diameter and pancreatic-gland width according to age. METHODS: Patients referred for either upper endoscopy or EUS for an indication unrelated to pancreaticobiliary disease were prospectively enrolled. Patients were stratified by age (<40, 40-60, >60 years). Each patient was assessed for the presence of EUS findings for chronic pancreatitis. Logistic regression was used to identify factors associated with an abnormality. RESULTS: A total of 120 patients (63 men, 57 women; median age, 52 years, interquartile range [IQR] 40-61 years) were prospectively evaluated. At least one parenchymal and/or ductular abnormality was identified in 28% of the patients, with a trend of increasing abnormality with age: <40 years (23%), 40 to 60 years (25%), and >60 years (39%); p = 0.13. No patient had more than 3 abnormal EUS features. Hyperechoic stranding (n = 22) was the most common finding in all age groups. The odds for any abnormality in men (relative to women) was significantly higher (OR 2.9: 95% CI[1.2, 6.8], p = 0.01), with 38% of men and 18% of women having an abnormality. Smoking, low alcohol intake, body mass index, and endoscopic finding were not significantly associated with an abnormal EUS. The overall median pancreatic-gland width and main pancreatic duct diameter were 15 mm (IQR 6-25 mm) and 1.7 mm (IQR 0.9-4.3 mm), respectively. CONCLUSIONS: The frequency of EUS abnormalities in patients without clinical evidence of chronic pancreatitis increases with age, particularly after 60 years of age. The threshold number of EUS criteria for the diagnosis of chronic pancreatitis is variable. However, the typically used standard of 3 or more criteria appears appropriate. A higher number of threshold criteria may be needed in males and to a lesser extent in patients over 40 years of age, which should be related to clinical history and other structural or functional studies. Ductal or parenchymal calculi, ductal narrowing, ductal dilatation, or more than 3 abnormalities appear to be more specific features for the EUS diagnosis of chronic pancreatitis at any age. PMID- 15758912 TI - High-dose allopurinol for prevention of post-ERCP pancreatitis: a prospective randomized double-blind controlled trial. AB - BACKGROUND: Pancreatitis is the most common major complication of diagnostic and therapeutic ERCP. Allopurinol, a xanthine oxidase inhibitor that blocks generation of oxygen-derived free radicals, potentially may prevent post-ERCP pancreatitis. This study assessed the efficacy of high-dose oral allopurinol for prevention of post-ERCP pancreatitis. METHODS: A prospective, double-blind, placebo-controlled trial was conducted in 250 patients undergoing ERCP. Patients were randomized to receive allopurinol (600 mg) or placebo orally at 15 and 3 hours before the procedure. Patients were clinically evaluated, and serum amylase levels were determined before ERCP and at 6 and 24 hours thereafter. Standardized criteria were used to diagnose and to grade the severity of post-ERCP pancreatitis. RESULTS: A total of 243 patients were included in the analysis. The two groups were similar with regard to age; gender; underlying disease; indication for treatment; ERCP findings; and type of treatment, except for biliary sphincterotomy. Only 43 patients in the allopurinol group underwent biliary sphincterotomy vs. 87 in the placebo group ( p < 0.001). The frequency of acute pancreatitis was significantly lower in the allopurinol vs. the placebo group in the final multinomial regression analysis: allopurinol group, 4/125 (3.2%), with all 4 cases graded as mild, vs. placebo group, 21/118 (17.8%), of which 8/118 (6.8%) were graded as mild, 11/118 (9.3%) as moderate, and 2/118 (1.6%) as severe with fatal outcome ( p < 0.001). The protective effect of allopurinol was also apparent in the diagnostic ERCP and the biliary sphincterotomy subgroups when the frequency of post-ERCP pancreatitis was analyzed after stratification by procedure. The mean duration of hospitalization for pancreatitis was significantly shorter in the allopurinol compared with the placebo group (2.5 vs. 5.67 days; p < 0.001). CONCLUSIONS: Pretreatment with high dose, orally administered allopurinol decreases the frequency of post-ERCP pancreatitis. Despite the promising results of this prospective, randomized trial, further studies are needed to verify these observations before allopurinol can be recommended for routine clinical use. PMID- 15758913 TI - The utility of EUS-guided FNA in the diagnosis of metastatic breast cancer to the esophagus and the mediastinum. AB - BACKGROUND: Breast cancer can metastasize to the esophagus and the mediastinum. EUS-guided FNA (EUS-FNA) is being used increasingly as a less invasive alternative to mediastinoscopy for procuring a tissue diagnosis of mediastinal disease and may be useful for the diagnosis of breast cancer metastatic to the esophagus and the mediastinum. METHODS: Twelve women (age range 54-82 years) with a history of breast cancer presented with dysphagia or other symptoms between 1 and 15 years after initial diagnosis and treatment. CT and endoscopy with biopsies suggested a mediastinal mass or lymphadenopathy with extrinsic esophageal compression but failed to provide a tissue diagnosis. EUS-FNA was performed for diagnosis. RESULTS: Cytologic evaluation of specimens obtained by EUS-FNA confirmed breast cancer metastases in 11 of 12 patients (91%). Recurrent disease was found in intramural masses and periesophageal lymph nodes. No complication resulted from any EUS-FNA procedure. CONCLUSIONS: EUS-FNA is safe and effective for the diagnosis of breast cancer metastases to the esophagus and the mediastinum. EUS-FNA may be useful as a first-line method of evaluation when breast cancer metastasis to the esophagus and the mediastinum is suspected. PMID- 15758915 TI - Third-tier fellowship training: advantages and program selection. PMID- 15758914 TI - Endoscopy or surgery for malignant GI outlet obstruction? AB - BACKGROUND: The treatment of gastroduodenal outflow obstruction (GOO) caused by malignant diseases represents a significant challenge. Open surgical gastrojejunostomy (GJ) has been the treatment of choice, but it has high morbidity and mortality rates. More recently, endoscopic placement of self expanding metallic stents (SEMS) has been proposed and the results of small, preliminary studies are encouraging. This study compared technical and clinical success, morbidity, mortality, and hospital stay in patients undergoing endoscopic and surgical treatment of GOO. METHODS: Medical records of 60 consecutive patients with GOO seen between April 1997 and November 2002 were retrospectively reviewed. Because of extremely short life expectancy, 13 patients were treated by insertion of a double-lumen nasogastric-jejunal tube. The remaining 47 patients (28 men, 19 women; mean age 73.5 years, range 48-92 years) with unresectable pancreatic (33), gastric (7), metastatic lymph nodal (4), papillary (2), and biliary (1) tumors were treated by placement of a SEMS (24) or open surgical GJ (23). RESULTS: The technical success rates were similar, but clinical success was lower in the GJ group (92% vs. 56%, p = 0.0067). The SEMS group had a shorter length of hospital stay (3.0 [1.4] days vs. 24.1 [10.3], p < 0.001). Thirty-day mortality was 30% in the GJ group, and 0% in the SEMS group ( p = 0.004). Morbidity was higher in the GJ compared with the SEMS group (61% vs. 17%, p = 0.0021). Mean survival was longer in the SEMS group (96.1 [9.6] days vs. 70.2 [36.2] days, p = 0.0165 for a single test of hypothesis; Bonferroni correction for a multiple testing removes this significance), consequently, out of-hospital survival was longer for the SEMS group (93.2 [9.3] days vs. 46.0 [31.5] days, p < 0.001). None of the endoscopic procedures required the assistance of an anesthesiologist or the use of an operating room. CONCLUSIONS: The results of this retrospective study suggest that SEMS insertion is better than surgical GJ for palliation of patients with GOO in terms of clinical success, morbidity, and mortality. Technical success rates were similar. SEMS placement should be proposed as the first-line treatment for relief of GOO. However, a randomized, comparative, prospective study of SEMS vs. laparoscopic GJ is needed. PMID- 15758916 TI - Pedunculated leiomyoma. PMID- 15758917 TI - Jejunal adenocarcinoma. PMID- 15758918 TI - Endoscopic visualization of an inverted appendix. PMID- 15758919 TI - Colocolic intussusception. PMID- 15758920 TI - Rectal endometriosis. PMID- 15758921 TI - Magnification endoscopy. PMID- 15758923 TI - Peroral transgastric endoscopic ligation of fallopian tubes with long-term survival in a porcine model. AB - BACKGROUND: We have previously reported the feasibility and safety of the peroral transgastric endoscopic approach for diagnostic peritoneoscopy, liver biopsy, and gastrojejunostomy with long-term survival in a porcine model. This approach eliminates incisions of the abdominal wall, providing a less invasive alternative to diagnostic and therapeutic laparoscopy. We now report successful performance of peroral endoscopic transgastric ligation of Fallopian tubes with long-term survival in a porcine model. METHODS: Six female 50-kg pigs had general anesthesia and irrigation of the stomach with an antibiotic solution. Gastric puncture was performed with needleknife electrocautery followed by balloon dilatation of the tract with 20-mm TTS dilating balloon (Microvasive). A standard upper endoscope that underwent high-level disinfection and gas sterilzation was advanced into the peritoneal cavity through a sterile overtube. Both Fallopian tubes were identified and one was ligated using Olympus Endoloops. The other patent tube served as a control. Tubal patency was evaluated by hysterosalpingogram before and after ligation. After a follow-up period of 2-3 weeks, the pigs were sacrificed for postmortem examination. RESULTS: The Fallopian tubes were easily accessed, identified and ligated in all 6 pigs. In each pig, fluoroscopy confirmed complete obstruction of the ligated tube with preserved patency of the other tube. All pigs survived well and ate heartily without any ill-effects. Postmortem examination did not reveal any peritonitis or intra-abdominal adhesions. The Endoloops were in place with complete obstruction of the ligated tubes and patency of the controls. Histopathologic examination of the tubes showed chronic inflammatory infiltrates without abscesses. CONCLUSIONS: The peroral endoscopic transgastric approach to ligation of the Fallopian tubes with long-term survival is technically feasible and safe in a porcine model. The endoscopic transgastric approach to the peritoneal cavity has potential for a wide array of diagnostic and therapeutic procedures. PMID- 15758924 TI - Gastroenterologists as surgeons: what they need to know. PMID- 15758925 TI - Temporary gastric electrical stimulation with orally or PEG-placed electrodes in patients with drug refractory gastroparesis. AB - BACKGROUND: Gastric electrical stimulation (GES) has been shown to be efficacious for drug refractory gastroparesis, but GES requires surgery. Placement of temporary GES electrodes endoscopically (ENDOstim) or via a PEG (PEGstim) is feasible, thereby allowing rapid assessment and comparison of temporary use (TEMP) with permanent (PERM) implantation. METHODS: Twenty consecutive patients with gastroparesis had TEMP electrodes placed (6 ENDOstim, 14 PEGstim). TEMP alone and TEMP vs. PERM placement of GES devices in 13 of 20 patients were compared via the following: average vomiting frequency score (VFS), total symptom score, days to symptom improvement, electrode impedance, and gastric emptying test. RESULTS: For patients receiving TEMP, GES demonstrated a rapid, significant, and sustained improvement in VFS, results similar to those for PERM. CONCLUSIONS: Both ENDO and PEG placement of GES electrodes are safe and effective in patients with gastroparesis, with outcomes that correspond to those achieved with permanent GES implantation. PMID- 15758926 TI - Wire-guided endoscopic snare papillectomy for tumors of the major duodenal papilla. AB - BACKGROUND: Endoscopic excision for adenoma of the major duodenal papilla was introduced as an alternative to surgery, but postprocedure pancreatitis is a serious drawback. This study assessed the feasibility and the safety of endoscopic papillectomy with a guidewire and pancreatic-duct stent insertion to prevent pancreatitis. METHODS: Six patients were enrolled. The snare loop was passed over a guidewire that had been inserted into the pancreatic duct. Immediately after snare resection, a pancreatic stent was placed along the indwelling guidewire. RESULTS: En bloc papillectomy and pancreatic stent insertion were performed successfully in all patients. Pancreatitis did not develop acutely in any patient. Complications included cholangitis (n = 1) and late-onset pancreatitis owing to the pancreatic stent (n = 1). Scant residual adenomatous tissue was present at resection margins in two patients and was treated endoscopically. CONCLUSIONS: Wire-guided endoscopic snare papillectomy in selected patients is a useful technique that maintains pancreatic-duct access for stent placement. This appears to prevent pancreatitis and to improve the outcome for patients undergoing endoscopic resection of papillary tumors. PMID- 15758928 TI - Successful recanalization of a completely obliterated esophageal stricture by using an endoscopic rendezvous maneuver. PMID- 15758927 TI - EUS-guided trucut biopsy in establishing autoimmune pancreatitis as the cause of obstructive jaundice. AB - BACKGROUND: The diagnosis of autoimmune pancreatitis can be difficult and often requires a larger specimen than can be provided by FNA alone to determine if the tissue sample obtained with EUS trucut biopsy (TCB) is sufficient to allow adequate histologic review to establish the diagnosis of autoimmune pancreatitis. METHODS: EUS TCB was performed in patients presenting with obstructive jaundice who were suspected of having autoimmune pancreatitis based on their clinical, laboratory and imaging studies. The charts were retrospectively reviewed to determine the feasibility of TCB. RESULTS: Between August 2002 and June 2004, 3 patients with obstructive jaundice and suspected autoimmune pancreatitis (AIP) underwent EUS TCB. In each case, a diagnosis of pancreatic cancer also was considered, and surgical resection was the planned therapy before the patient underwent EUS TCB. Histologic review of the TCB specimens established the diagnosis of AIP in two patients and identified nonspecific changes of chronic pancreatitis in the third patient. EUS-guided FNA was performed in two of the 3 patients and failed to establish the diagnosis in either patient. Other than mild transient abdominal pain (n = 1), no complications were identified. CONCLUSIONS: This preliminary study suggests that EUS TCB can safely establish the diagnosis of AIP. Doing so helps guide management and may help to avoid unnecessary surgery. Prospective studies are needed to verify these findings and to more clearly define the role of EUS TCB in these patients. PMID- 15758929 TI - Adenomyoma of the minor duodenal papilla. PMID- 15758930 TI - A magnified endoscopic view of esophageal melanocytosis. PMID- 15758931 TI - Retrograde intussusception caused by a sigmoid colonic tumor: reduction by the use of a transanal drainage tube. PMID- 15758932 TI - Esophageal melanocytosis in the setting of Addison's disease. PMID- 15758933 TI - Life-threatening pericarditis after N-butyl-2-cyanoacrylate injection for esophageal variceal bleeding: Case report. PMID- 15758934 TI - Diagnosis by EUS trucut biopsy of extrapulmonary tuberculosis in a patient with Crohn's disease treated with infliximab. PMID- 15758935 TI - Placement of a jejunal enteral tube through a percutaneous endoscopic jejunostomy to prevent recurrent aspiration during intestinal feeding. PMID- 15758936 TI - Signet-ring-cell adenocarcinoma arising from a hyperplastic polyp in the stomach. PMID- 15758937 TI - EUS-guided drainage of hepatic abscess. PMID- 15758938 TI - Giant esophageal papilloma. PMID- 15758939 TI - Why did colonic abscess occur? PMID- 15758940 TI - Ketamine for pediatric endoscopic procedures and as a sedation complement for adult patients. PMID- 15758941 TI - Balloon dilation in patients with diffuse esophageal spasm. PMID- 15758943 TI - Protective preconditioning by transient global ischemia in the rat: components of delayed injury progression and lasting protection distinguished by comparisons of depolarization thresholds for cell loss at long survival times. AB - Robust ischemic preconditioning has been shown in rodent brain, but there are concerns regarding the persistence of neuron protection. This issue was examined in rat hippocampus following 4-vessel occlusion (4-VO) ischemia, using DC shifts characteristic of ischemic depolarization to reproducibly define insult severity. Preconditioning ischemia producing 2 to 3.5 mins depolarization was followed at intervals of 2, 5, or 7 days by test insults of varied duration, after which CA1 counts were obtained at 1, 2, 4, or 12 weeks. Neuron loss in naive animals increased with depolarization time longer than 4 mins regardless of postischemic survival interval. Preconditioning 2, 5, or 7 days before test insults prolonged the injury threshold evaluated at 1 week survival to 15, 9, or 6 mins, respectively, showing robust protection and a rapid decay of the protected state. However, by 2 weeks survival after preconditioning at a 2-day interval, the injury threshold dramatically regressed from 15 to 9 mins. Thereafter protection remained relatively stable through 1 month, but slight progression of neuron injury was evident at 3 months. Inflammatory responses were seen in both naive and preconditioned hippocampi throughout this interval, appropriate to the extent of neuron injury. These studies show distinct components of transient and lasting protection after ischemic preconditioning. Finally, it was found that ischemic depolarization was delayed by approximately 1 min in optimally preconditioned rat hippocampus, in contrast to previous results in the gerbil, identifying one specific mechanism by which insult severity is reduced in this model. PMID- 15758944 TI - NFkappaB activates in vivo the synthesis of inducible Cox-2 in the brain. AB - Interleukin-1beta (IL-1beta) induces cyclooxygenase-2 (Cox-2) expression in many of its cellular targets resulting in production and release of prostaglandins. Although IL-1beta-induced Cox-2 expression most likely requires activation of nuclear transcription factor kappa B (NFkappaB) pathway, this has never been formally demonstrated in vivo. We tested this using a specific inhibitor of NFkappaB activation, the NEMO binding domain (NBD) peptide, that has been shown previously to be effective in various in vivo models of acute inflammation. Incubation of rat glioma cells with the NBD peptide blocked IL-1beta-induced NFkappaB nuclear translocation. Furthermore, after injection of a biotinylated version of the NBD peptide into the lateral ventricle of the brain, we found that it readily diffused to its potential cellular targets in vivo. To test the effects of the peptide on NFkappaB activation and Cox-2 expression in the brain, we injected it intracerebroventricularly (36 microg/rat) into rats before intraperitoneal injection of IL-1beta (60 microg/kg). Treatment with NBD peptide completely abolished IL-1beta-induced NFkappaB activation and Cox-2 synthesis in microvasculature. In contrast, the peptide had no effect on constitutive neuronal Cox-2. These findings strongly support the hypothesis that IL-1beta-induced NFkappaB activation plays a major role in transmission of immune signals from the periphery to the brain. PMID- 15758945 TI - Estrogen restores postischemic sensitivity to the thromboxane mimetic U46619 in rat pial artery. AB - The objectives of the study were to (1) characterize the dose-response relationship to the TXA2 analog, U46619 (0.01, 0.1, and 1 micromol/L) after global cerebral ischemia, (2) determine whether chronic 17beta-estradiol (E2) replacement alters this relationship, and (3) determine if E2's mechanisms are transduced through cognate estrogen receptors. Rats were assigned to five groups (n=6): placebo-implanted ovariectomized (OVX) females, OVX plus chronic E2 (CE), OVX plus acute E2 (AE), OVX plus chronic E2 plus the estrogen receptor inhibitor ICI 182,780 (CEI), and OVX plus acute E2 plus ICI 182,780 (AEI). Rats were anesthetized, intubated, cannulated (femoral artery and vein), fitted with a closed cranial window, and subjected to 15-min reversible forebrain ischemia (4 vessel occlusion, 4-VO) and 60 mins of reperfusion. Arterial blood gases, intrawindow pressure, and temperature were controlled. Vessel diameter was measured before and 5 mins after superfusion of each concentration of U46619. Compared with preischemic responses, contractile response to U46619 was depressed at all concentrations after ischemia in the OVX group. In the chronic E2 and acute E2 groups, contractile response to 1 micromol/L of U46619 was normalized to near baseline values. However, in the CEI and the AEI groups, postischemic vasoconstriction was similar to that observed in the OVX rats. We conclude that E2 targets the cerebral microvasculature to preserve postischemic pial artery reactivity and that the effect is receptor mediated. Restoration of normal constriction to vascular agonists may be an important mechanism by which E2 protects the vasculature and diminishes tissue damage after ischemia. PMID- 15758946 TI - Sleep-related brain activation does not increase the permeability of the blood brain barrier to glucose. AB - We compared blood-brain barrier (BBB) permeability to glucose between quiet wakefulness and rapid-eye-movement (REM) sleep to assess whether changes in BBB permeability play a role in coupling glucose supply to the physiologic metabolic needs of the brain. Male Sprague-Dawley rats were prepared with electrodes for wake-sleep state scoring and with arterial and venous catheters. Using the single pass, dual-label indicator method, unidirectional glucose extraction by the brain and cerebral blood flow (CBF) were simultaneously measured during states of quiet wakefulness (n=12) or REM sleep (n=7). The product of BBB surface area and permeability to glucose (PS product) was computed in each state. During REM sleep, CBF significantly exceeded that during quiet wakefulness in all regions but the cerebellum, whereas the difference in the PS product between quiet wakefulness and REM sleep was not statistically significant in any brain region. In the brain as a whole, CBF significantly increased 29% from quiet wakefulness to REM sleep, while a nonsignificant 0.8% increase occurred in the PS product. During REM sleep, the increase in CBF indicates a higher rate of brain glucose consumption than in quiet wakefulness, given the tight flow-metabolism coupling in the brain. Therefore, these data show that modulation of BBB permeability to glucose is not a mechanism that provides 'energy on demand' during the physiologic brain activation characterising REM sleep. PMID- 15758948 TI - Acute intravenous low- and high-dose cocaine reduces quantitative global and regional cerebral blood flow in recently abstinent subjects with cocaine use disorder. AB - Cocaine-induced hypoperfusion, a risk factor for ischemic stroke, has not been fully characterized during experimental drug-taking among individuals with cocaine use disorder. We sought to examine cocaine's dose-dependent, time-related effects on cerebral blood flow. In a double-blind, randomized human laboratory study with a counterbalanced order of drug administration, 31 male and female subjects with cocaine use disorder were divided into two groups receiving either (a) low-dose cocaine (0.325 mg/kg intravenously) or placebo (N=15) or (b) high dose cocaine (0.650 mg/kg intravenously) or placebo (N=16). The different dose conditions were administered on test days separated by a rest period of >or=48 h. Cerebral blood flow was assessed quantitatively using H(2)O(15) positron emission tomography. Experimentally administered low- and high-dose cocaine conditions versus their corresponding placebo conditions were associated with global and regional hypoperfusion. The trend for high- versus low-dose cocaine to be associated with greater hypoperfusion achieved statistical significance only for the dopamine-rich sublobar and midbrain regions. Cocaine's hypoperfusion effects were maximal at 8 mins after infusion (i.e., at about the expected peak of intravenous cocaine levels) and had mostly dissipated by 32 mins after infusion. Although hypoperfusion occurred throughout the brain, the left hemispheric dopamine-rich sublobar region was the most severely affected. Cocaine-induced cerebral hypoperfusion is associated with the time course of its pharmacological effects, and dopamine-rich areas, particularly in the left hemisphere, may be most vulnerable. Increasingly larger doses of cocaine may be associated with greater risk for ischemic stroke. PMID- 15758947 TI - General anesthesia improves fetal cerebral oxygenation without evidence of subsequent neuronal injury. AB - Anesthetic exposure during pregnancy is viewed as a relatively routine medical practice. However, recent rodent studies have suggested that common anesthetic agents can damage the developing brain. Here we assessed this claim in a higher order species by exposing previously instrumented near-term pregnant sheep at gestational day 122 (+/-1) to a combination of midazolam, sodium thiopental, and isoflurane at clinically relevant doses and means of anesthetic delivery (i.e., active ventilation). Four hours of maternal general anesthesia produced an initial increase in fetal systemic oxygenation and a sustained increase in fetal cerebral oxygenation, as determined by in utero near-infrared spectroscopy. Postexposure monitoring failed to identify changes in physiologic status that could be injurious to the fetal brain. Finally, through the histologic assessment of noninstrumented sheep at the same gestational time point, we found no evidence for a direct fetal neuro-toxic effect of our triple-drug regimen. Collectively, these results appear to corroborate the presumed safety of inhalational anesthetic use during pregnancy. PMID- 15758949 TI - Measurements of BOLD/CBV ratio show altered fMRI hemodynamics during stroke recovery in rats. AB - Brain responses to external stimuli after permanent and transient ischemic insults have been documented using cerebral blood volume weighted (CBVw) functional magnetic resonance imaging (fMRI) in correlation with tissue damage and neurological recovery. Here, we extend our previous studies of stroke recovery in rat models of focal cerebral ischemia by comparing blood oxygen level dependent (BOLD) and cerebral blood volume (CBV) changes. Responses to forepaw stimulation were measured in normal rats (n=5) and stroke rats subjected to 2 h of middle cerebral artery occlusion (n=6). Functional magnetic resonance imaging was performed 2 weeks after stroke to evaluate the recovery process. After stroke, animals showed variable degrees of fMRI activation in ipsilesional cortex, the extent of which did not correlate with structural damages as measured using apparent diffusion coefficient, fractional anisotropy, blood volume, and vessel size index. While the contralesional cortex showed good overlap between BOLD and CBV-activated regions, the ipsilesional cortex showed low covariance between significantly activated voxels by BOLD and CBVw techniques. In particular, the relative activation during contralateral stimuli in the ipsilesional somatosensory cortex was significantly higher for CBVw responses than BOLD, which might be due to stroke-related alterations in fMRI hemodynamic coupling. Aberrant subcortical activations were also observed. When unaffected forelimbs were stimulated, strong bilateral responses were observed. However, little thalamic responses accompanied stimulation of affected forelimbs despite significant activation in the ipsilesional somatosensory cortex. These results suggest that stroke affects not only local hemodynamics and coupling but also other factors including neural connectivity. PMID- 15758950 TI - Long-term in vivo investigation of mouse cerebral microcirculation by fluorescence confocal microscopy in the area of focal ischemia. AB - This study was designed to assess that mouse pial and cortical microcirculation can be monitored in the long term directly in the area of focal ischemia, using in vivo fluorescence microscopy. A closed cranial window was placed over the left parieto-occipital cortex of C57BL/6J mice. Local microcirculation was recorded in real time through the window using laser-scanning confocal fluorescence microscopy after intravenous injection of fluorescent erythrocytes and dextran. The basal velocity of erythrocytes through intraparenchymal capillaries was 0.53+/-0.30 mm/sec (n=121 capillaries in 10 mice). Two branches of the middle cerebral artery were topically cauterized through the window. Blood flow evaluated by laser-Doppler flowmetry in two distinct areas indicated the occurrence of an ischemic core (15.2%+/-5.9% of baseline for at least 2 h) and a penumbral zone. Magnetic resonance imaging and histology were used to characterize the ischemic area at 24 h after occlusion. The infarct volume was 7.3+/-3.2 mm(3) (n=6). Microcirculation was repeatedly videorecorded using fluorescence confocal microscopy over the next month. After the decrease following arterial occlusion, capillary erythrocyte velocity was significantly higher than baseline 1 week later, and attained 0.74+/-0.51 mm/sec (n=76 capillaries in six mice, P<0.005) after 1 month, while venous and capillary network remodeling was assessed, with a marked decrease in tortuosity. Immunohistochemistry revealed a zone of necrotic tissue into the infarct epicenter, with activated astrocytes at its border. Such long-term investigations in ischemic cortex brings new insight into the microcirculatory changes induced by focal ischemia and show the feasibility of long-term fluorescence studies in the mouse cortex. PMID- 15758951 TI - Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones. AB - Ion channels formed by the TRP (transient receptor potential) superfamily of proteins act as sensors for temperature, osmolarity, mechanical stress and taste. The growth cones of developing axons are responsible for sensing extracellular guidance factors, many of which trigger Ca2+ influx at the growth cone; however, the identity of the ion channels involved remains to be clarified. Here, we report that TRP-like channel activity exists in the growth cones of cultured Xenopus neurons and can be modulated by exposure to netrin-1 and brain-derived neurotrophic factor, two chemoattractants for axon guidance. Whole-cell recording from growth cones showed that netrin-1 induced a membrane depolarization, part of which remained after all major voltage-dependent channels were blocked. Furthermore, the membrane depolarization was sensitive to blockers of TRP channels. Pharmacological blockade of putative TRP currents or downregulation of Xenopus TRP-1 (xTRPC1) expression with a specific morpholino oligonucleotide abolished the growth-cone turning and Ca2+ elevation induced by a netrin-1 gradient. Thus, TRPC currents reflect early events in the growth cone's detection of some extracellular guidance signals, resulting in membrane depolarization and cytoplasmic Ca2+ elevation that mediates the turning of growth cones. PMID- 15758952 TI - Essential role of TRPC channels in the guidance of nerve growth cones by brain derived neurotrophic factor. AB - Brain-derived neurotrophic factor (BDNF) is known to promote neuronal survival and differentiation and to guide axon extension both in vitro and in vivo. The BDNF-induced chemo-attraction of axonal growth cones requires Ca2+ signalling, but how Ca2+ is regulated by BDNF at the growth cone remains largely unclear. Extracellular application of BDNF triggers membrane currents resembling those through TRPC (transient receptor potential canonical) channels in rat pontine neurons and in Xenopus spinal neurons. Here, we report that in cultured cerebellar granule cells, TRPC channels contribute to the BDNF-induced elevation of Ca2+ at the growth cone and are required for BDNF-induced chemo-attractive turning. Several members of the TRPC family are highly expressed in these neurons, and both Ca2+ elevation and growth-cone turning induced by BDNF are abolished by pharmacological inhibition of TRPC channels, overexpression of a dominant-negative form of TRPC3 or TRPC6, or downregulation of TRPC3 expression via short interfering RNA. Thus, TRPC channel activity is essential for nerve growth-cone guidance by BDNF. PMID- 15758954 TI - Bad faith at Los Alamos. PMID- 15758953 TI - Conserved modes of recruitment of ATM, ATR and DNA-PKcs to sites of DNA damage. AB - Ataxia-telangiectasia mutated (ATM), ataxia-telangiectasia and Rad3-related (ATR) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) are members of the phosphoinositide-3-kinase-related protein kinase (PIKK) family, and are rapidly activated in response to DNA damage. ATM and DNA-PKcs respond mainly to DNA double-strand breaks, whereas ATR is activated by single-stranded DNA and stalled DNA replication forks. In all cases, activation involves their recruitment to the sites of damage. Here we identify related, conserved carboxy-terminal motifs in human Nbs1, ATRIP and Ku80 proteins that are required for their interaction with ATM, ATR and DNA-PKcs, respectively. These motifs are essential not only for efficient recruitment of ATM, ATR and DNA-PKcs to sites of damage, but are also critical for ATM-, ATR- and DNA-PKcs-mediated signalling events that trigger cell cycle checkpoints and DNA repair. Our findings reveal that recruitment of these PIKKs to DNA lesions occurs by common mechanisms through an evolutionarily conserved motif, and provide direct evidence that PIKK recruitment is required for PIKK-dependent DNA-damage signalling. PMID- 15758955 TI - Killers on the loose. PMID- 15758956 TI - NASA's funding shortfall means journey's end for Voyager probes. PMID- 15758957 TI - Anthropologists walk tall after unearthing hominid. PMID- 15758958 TI - Senate resignations mar academy's birthday. PMID- 15758959 TI - Gene-therapy trials to restart following cancer risk review. PMID- 15758960 TI - Career charter sets out rights of Europe's young scientists. PMID- 15758961 TI - Biosafety law brings stem-cell research to Brazil. PMID- 15758962 TI - Climatologists seek clear view of Asia's smog. PMID- 15758963 TI - Science agencies get fresh paymasters in Republican revamp. PMID- 15758964 TI - Police urge speedy action to clean up home drug factories. PMID- 15758966 TI - Counterfeit pharmaceuticals: murder by medicine. PMID- 15758967 TI - Counterfeit pharmaceuticals: in the line of fire. Dora Akunyili interviewed by Peter Aldhous. PMID- 15758968 TI - Brown knew particles were smaller than pollen. PMID- 15758969 TI - Concern over deep-sea reefs is widespread. PMID- 15758970 TI - HIV drug remains unproven without placebo trial. PMID- 15758976 TI - Eggs alone. PMID- 15758977 TI - Biodiversity: fossils make waves. PMID- 15758978 TI - Astronomy: stellar mass limited. PMID- 15758980 TI - Epigenetics: surveillance team against cancer. PMID- 15758981 TI - Structural biology: methanol maker. PMID- 15758982 TI - Semiconductor technology: negatively successful. PMID- 15758984 TI - Planetary sciences: a smashing pair. PMID- 15758983 TI - Developmental biology: sperm-egg fusion unscrambled. PMID- 15758985 TI - Animal behaviour: meals sized up. PMID- 15758986 TI - Palaeobiology: dating earliest life. PMID- 15758988 TI - Planetary science: Meteor Crater formed by low-velocity impact. AB - Meteor Crater in Arizona was the first terrestrial structure to be widely recognized as a meteorite impact scar and has probably been more intensively studied than any other impact crater on Earth. We have discovered something surprising about its mode of formation--namely that the surface-impact velocity of the iron meteorite that created Meteor Crater was only about 12 km s(-1). This is close to the 9.4 km s(-1) minimum originally proposed but far short of the 15 20 km s(-1) that has been widely assumed--a realization that clears up a long standing puzzle about why the crater does not contain large volumes of rock melted by the impact. PMID- 15758989 TI - Musical imagery: sound of silence activates auditory cortex. AB - Auditory imagery occurs when one mentally rehearses telephone numbers or has a song 'on the brain'--it is the subjective experience of hearing in the absence of auditory stimulation, and is useful for investigating aspects of human cognition. Here we use functional magnetic resonance imaging to identify and characterize the neural substrates that support unprompted auditory imagery and find that auditory and visual imagery seem to obey similar basic neural principles. PMID- 15758990 TI - Imaging of Titan from the Cassini spacecraft. AB - Titan, the largest moon of Saturn, is the only satellite in the Solar System with a substantial atmosphere. The atmosphere is poorly understood and obscures the surface, leading to intense speculation about Titan's nature. Here we present observations of Titan from the imaging science experiment onboard the Cassini spacecraft that address some of these issues. The images reveal intricate surface albedo features that suggest aeolian, tectonic and fluvial processes; they also show a few circular features that could be impact structures. These observations imply that substantial surface modification has occurred over Titan's history. We have not directly detected liquids on the surface to date. Convective clouds are found to be common near the south pole, and the motion of mid-latitude clouds consistently indicates eastward winds, from which we infer that the troposphere is rotating faster than the surface. A detached haze at an altitude of 500 km is 150-200 km higher than that observed by Voyager, and more tenuous haze layers are also resolved. PMID- 15758991 TI - Experimental one-way quantum computing. AB - Standard quantum computation is based on sequences of unitary quantum logic gates that process qubits. The one-way quantum computer proposed by Raussendorf and Briegel is entirely different. It has changed our understanding of the requirements for quantum computation and more generally how we think about quantum physics. This new model requires qubits to be initialized in a highly entangled cluster state. From this point, the quantum computation proceeds by a sequence of single-qubit measurements with classical feedforward of their outcomes. Because of the essential role of measurement, a one-way quantum computer is irreversible. In the one-way quantum computer, the order and choices of measurements determine the algorithm computed. We have experimentally realized four-qubit cluster states encoded into the polarization state of four photons. We characterize the quantum state fully by implementing experimental four-qubit quantum state tomography. Using this cluster state, we demonstrate the feasibility of one-way quantum computing through a universal set of one- and two qubit operations. Finally, our implementation of Grover's search algorithm demonstrates that one-way quantum computation is ideally suited for such tasks. PMID- 15758992 TI - Structural basis of HutP-mediated anti-termination and roles of the Mg2+ ion and L-histidine ligand. AB - HutP regulates the expression of the hut structural genes of Bacillus subtilis by an anti-termination mechanism and requires two components, Mg2+ ions and L histidine. HutP recognizes three UAG triplet units, separated by four non conserved nucleotides on the terminator region. Here we report the 1.60-A resolution crystal structure of the quaternary complex (HutP-L-histidine-Mg2+-21 base single-stranded RNA). In the complex, the RNA adopts a novel triangular fold on the hexameric surface of HutP, without any base-pairing, and binds to the protein mostly by specific protein-base interactions. The structure explains how the HutP and RNA interactions are regulated critically by the l-histidine and Mg2+ ion through the structural rearrangement. To gain insights into these structural rearrangements, we solved two additional crystal structures (uncomplexed HutP and HutP-L-histidine-Mg2+) that revealed the intermediate structures of HutP (before forming an active structure) and the importance of the Mg2+ ion interactions in the complexes. PMID- 15758993 TI - An upper limit to the masses of stars. AB - There is no accepted upper mass limit for stars. Such a basic quantity eludes both theory and observation, because of an imperfect understanding of the star formation process and because of incompleteness in surveying the Galaxy. The Arches cluster is ideal for investigating such limits, being large enough to expect stars at least as massive as approximately 500 solar masses (approximately 500 Mo; based on a typical mass function), and young enough for its most massive members to still be visible. It is also old enough to be free of its natal molecular cloud, it is at a well-established distance, and it is close enough for us to discern its individual stars. Here I report an absence of stars with initial masses greater than 130 Mo in the Arches cluster, whereas the typical mass function predicts 18. I conclude that this indicates a firm limit of 150 Mo for stars; the probability that the observations are consistent with there being no upper limit is 10(-8). PMID- 15758994 TI - General observation of n-type field-effect behaviour in organic semiconductors. AB - Organic semiconductors have been the subject of active research for over a decade now, with applications emerging in light-emitting displays and printable electronic circuits. One characteristic feature of these materials is the strong trapping of electrons but not holes: organic field-effect transistors (FETs) typically show p-type, but not n-type, conduction even with the appropriate low work-function electrodes, except for a few special high-electron-affinity or low bandgap organic semiconductors. Here we demonstrate that the use of an appropriate hydroxyl-free gate dielectric--such as a divinyltetramethylsiloxane bis(benzocyclobutene) derivative (BCB; ref. 6)--can yield n-channel FET conduction in most conjugated polymers. The FET electron mobilities thus obtained reveal that electrons are considerably more mobile in these materials than previously thought. Electron mobilities of the order of 10(-3) to 10(-2) cm(2) V( 1) s(-1) have been measured in a number of polyfluorene copolymers and in a dialkyl-substituted poly(p-phenylenevinylene), all in the unaligned state. We further show that the reason why n-type behaviour has previously been so elusive is the trapping of electrons at the semiconductor-dielectric interface by hydroxyl groups, present in the form of silanols in the case of the commonly used SiO2 dielectric. These findings should therefore open up new opportunities for organic complementary metal-oxide semiconductor (CMOS) circuits, in which both p type and n-type behaviours are harnessed. PMID- 15758995 TI - Ultrafast memory loss and energy redistribution in the hydrogen bond network of liquid H2O. AB - Many of the unusual properties of liquid water are attributed to its unique structure, comprised of a random and fluctuating three-dimensional network of hydrogen bonds that link the highly polar water molecules. One of the most direct probes of the dynamics of this network is the infrared spectrum of the OH stretching vibration, which reflects the distribution of hydrogen-bonded structures and the intermolecular forces controlling the structural dynamics of the liquid. Indeed, water dynamics has been studied in detail, most recently using multi-dimensional nonlinear infrared spectroscopy for acquiring structural and dynamical information on femtosecond timescales. But owing to technical difficulties, only OH stretching vibrations in D2O or OD vibrations in H2O could be monitored. Here we show that using a specially designed, ultrathin sample cell allows us to observe OH stretching vibrations in H2O. Under these fully resonant conditions, we observe hydrogen bond network dynamics more than one order of magnitude faster than seen in earlier studies that include an extremely fast sweep in the OH frequencies on a 50-fs timescale and an equally fast disappearance of the initial inhomogeneous distribution of sites. Our results highlight the efficiency of energy redistribution within the hydrogen-bonded network, and that liquid water essentially loses the memory of persistent correlations in its structure within 50 fs. PMID- 15758996 TI - The formation of cubic ice under conditions relevant to Earth's atmosphere. AB - An important mechanism for ice cloud formation in the Earth's atmosphere is homogeneous nucleation of ice in aqueous droplets, and this process is generally assumed to produce hexagonal ice. However, there are some reports that the metastable crystalline phase of ice, cubic ice, may form in the Earth's atmosphere. Here we present laboratory experiments demonstrating that cubic ice forms when micrometre-sized droplets of pure water and aqueous solutions freeze homogeneously at cooling rates approaching those found in the atmosphere. We find that the formation of cubic ice is dominant when droplets freeze at temperatures below 190 K, which is in the temperature range relevant for polar stratospheric clouds and clouds in the tropical tropopause region. These results, together with heat transfer calculations, suggest that cubic ice will form in the Earth's atmosphere. If there were a significant fraction of cubic ice in some cold clouds this could increase their water vapour pressure, and modify their microphysics and ice particle size distributions. Under specific conditions this may lead to enhanced dehydration of the tropopause region. PMID- 15758997 TI - Measuring the onset of locking in the Peru-Chile trench with GPS and acoustic measurements. AB - The subduction zone off the west coast of South America marks the convergence of the oceanic Nazca plate and the continental South America plate. Nazca-South America convergence over the past 23 million years has created the 6-km-deep Peru Chile trench, 150 km offshore. High pressure between the plates creates a locked zone, leading to deformation of the overriding plate. The surface area of this locked zone is thought to control the magnitude of co-seismic release and is limited by pressure, temperature, sediment type and fluid content. Here we present seafloor deformation data from the submerged South America plate obtained from a combination of Global Positioning System (GPS) receivers and acoustic transponders. We estimate that the measured horizontal surface motion perpendicular to the trench is consistent with a model having no slip along the thrust fault between 2 and 40 km depth. A tsunami in 1996, 200 km north of our site, was interpreted as being the result of an anomalously shallow interplate earthquake. Seismic coupling at shallow depths, such as we observe, may explain why co-seismic events in the Peruvian subduction zone create large tsunamis. PMID- 15758998 TI - Cycles in fossil diversity. AB - It is well known that the diversity of life appears to fluctuate during the course of the Phanerozoic, the eon during which hard shells and skeletons left abundant fossils (0-542 million years ago). Here we show, using Sepkoski's compendium of the first and last stratigraphic appearances of 36,380 marine genera, a strong 62 +/- 3-million-year cycle, which is particularly evident in the shorter-lived genera. The five great extinctions enumerated by Raup and Sepkoski may be an aspect of this cycle. Because of the high statistical significance we also consider the contributions of environmental factors, and possible causes. PMID- 15758999 TI - Agricultural runoff fuels large phytoplankton blooms in vulnerable areas of the ocean. AB - Biological productivity in most of the world's oceans is controlled by the supply of nutrients to surface waters. The relative balance between supply and removal of nutrients--including nitrogen, iron and phosphorus--determines which nutrient limits phytoplankton growth. Although nitrogen limits productivity in much of the ocean, large portions of the tropics and subtropics are defined by extreme nitrogen depletion. In these regions, microbial denitrification removes biologically available forms of nitrogen from the water column, producing substantial deficits relative to other nutrients. Here we demonstrate that nitrogen-deficient areas of the tropical and subtropical oceans are acutely vulnerable to nitrogen pollution. Despite naturally high nutrient concentrations and productivity, nitrogen-rich agricultural runoff fuels large (54-577 km2) phytoplankton blooms in the Gulf of California. Runoff exerts a strong and consistent influence on biological processes, in 80% of cases stimulating blooms within days of fertilization and irrigation of agricultural fields. We project that by the year 2050, 27-59% of all nitrogen fertilizer will be applied in developing regions located upstream of nitrogen-deficient marine ecosystems. Our findings highlight the present and future vulnerability of these ecosystems to agricultural runoff. PMID- 15759000 TI - The global distribution of clinical episodes of Plasmodium falciparum malaria. AB - Interest in mapping the global distribution of malaria is motivated by a need to define populations at risk for appropriate resource allocation and to provide a robust framework for evaluating its global economic impact. Comparison of older and more recent malaria maps shows how the disease has been geographically restricted, but it remains entrenched in poor areas of the world with climates suitable for transmission. Here we provide an empirical approach to estimating the number of clinical events caused by Plasmodium falciparum worldwide, by using a combination of epidemiological, geographical and demographic data. We estimate that there were 515 (range 300-660) million episodes of clinical P. falciparum malaria in 2002. These global estimates are up to 50% higher than those reported by the World Health Organization (WHO) and 200% higher for areas outside Africa, reflecting the WHO's reliance upon passive national reporting for these countries. Without an informed understanding of the cartography of malaria risk, the global extent of clinical disease caused by P. falciparum will continue to be underestimated. PMID- 15759001 TI - Mediation of pathogen resistance by exudation of antimicrobials from roots. AB - Most plant species are resistant to most potential pathogens. It is not known why most plant-microbe interactions do not lead to disease, although recent work indicates that this basic disease resistance is multi-factorial. Here we show that the exudation of root-derived antimicrobial metabolites by Arabidopsis thaliana confers tissue-specific resistance to a wide range of bacterial pathogens. However, a Pseudomonas syringae strain that is both at least partly resistant to these compounds and capable of blocking their synthesis/exudation is able to infect the roots and cause disease. We also show that the ability of this P. syringae strain to block antimicrobial exudation is dependent on the type III secretory system. PMID- 15759002 TI - Spike-timing-dependent synaptic plasticity depends on dendritic location. AB - In the neocortex, each neuron receives thousands of synaptic inputs distributed across an extensive dendritic tree. Although postsynaptic processing of each input is known to depend on its dendritic location, it is unclear whether activity-dependent synaptic modification is also location-dependent. Here we report that both the magnitude and the temporal specificity of spike-timing dependent synaptic modification vary along the apical dendrite of rat cortical layer 2/3 pyramidal neurons. At the distal dendrite, the magnitude of long-term potentiation is smaller, and the window of pre-/postsynaptic spike interval for long-term depression (LTD) is broader. The spike-timing window for LTD correlates with the window of action potential-induced suppression of NMDA (N-methyl-D aspartate) receptors; this correlation applies to both their dendritic location dependence and pharmacological properties. Presynaptic stimulation with partial blockade of NMDA receptors induced LTD and occluded further induction of spike timing-dependent LTD, suggesting that NMDA receptor suppression underlies LTD induction. Computer simulation studies showed that the dendritic inhomogeneity of spike-timing-dependent synaptic modification leads to differential input selection at distal and proximal dendrites according to the temporal characteristics of presynaptic spike trains. Such location-dependent tuning of inputs, together with the dendritic heterogeneity of postsynaptic processing, could enhance the computational capacity of cortical pyramidal neurons. PMID- 15759003 TI - The receptors and coding logic for bitter taste. AB - The sense of taste provides animals with valuable information about the nature and quality of food. Bitter taste detection functions as an important sensory input to warn against the ingestion of toxic and noxious substances. T2Rs are a family of approximately 30 highly divergent G-protein-coupled receptors (GPCRs) that are selectively expressed in the tongue and palate epithelium and are implicated in bitter taste sensing. Here we demonstrate, using a combination of genetic, behavioural and physiological studies, that T2R receptors are necessary and sufficient for the detection and perception of bitter compounds, and show that differences in T2Rs between species (human and mouse) can determine the selectivity of bitter taste responses. In addition, we show that mice engineered to express a bitter taste receptor in 'sweet cells' become strongly attracted to its cognate bitter tastants, whereas expression of the same receptor (or even a novel GPCR) in T2R-expressing cells resulted in mice that are averse to the respective compounds. Together these results illustrate the fundamental principle of bitter taste coding at the periphery: dedicated cells act as broadly tuned bitter sensors that are wired to mediate behavioural aversion. PMID- 15759004 TI - Spatial bistability of Dpp-receptor interactions during Drosophila dorsal-ventral patterning. AB - In many developmental contexts, a locally produced morphogen specifies positional information by forming a concentration gradient over a field of cells. However, during embryonic dorsal-ventral patterning in Drosophila, two members of the bone morphogenetic protein (BMP) family, Decapentaplegic (Dpp) and Screw (Scw), are broadly transcribed but promote receptor-mediated signalling in a restricted subset of expressing cells. Here we use a novel immunostaining protocol to visualize receptor-bound BMPs and show that both proteins become localized to a sharp stripe of dorsal cells. We demonstrate that proper BMP localization involves two distinct processes. First, Dpp undergoes directed, long-range extracellular transport. Scw also undergoes long-range movement, but can do so independently of Dpp transport. Second, an intracellular positive feedback circuit promotes future ligand binding as a function of previous signalling strength. These data elicit a model in which extracellular Dpp transport initially creates a shallow gradient of BMP binding that is acted on by positive intracellular feedback to produce two stable states of BMP-receptor interactions, a spatial bistability in which BMP binding and signalling capabilities are high in dorsal-most cells and low in lateral cells. PMID- 15759006 TI - In the picture. PMID- 15759005 TI - The immunoglobulin superfamily protein Izumo is required for sperm to fuse with eggs. AB - Representing the 60 trillion cells that build a human body, a sperm and an egg meet, recognize each other, and fuse to form a new generation of life. The factors involved in this important membrane fusion event, fertilization, have been sought for a long time. Recently, CD9 on the egg membrane was found to be essential for fusion, but sperm-related fusion factors remain unknown. Here, by using a fusion-inhibiting monoclonal antibody and gene cloning, we identify a mouse sperm fusion-related antigen and show that the antigen is a novel immunoglobulin superfamily protein. We have termed the gene Izumo and produced a gene-disrupted mouse line. Izumo-/- mice were healthy but males were sterile. They produced normal-looking sperm that bound to and penetrated the zona pellucida but were incapable of fusing with eggs. Human sperm also contain Izumo and addition of the antibody against human Izumo left the sperm unable to fuse with zona-free hamster eggs. PMID- 15759007 TI - Come together. PMID- 15759010 TI - Heregulin-beta1 regulates the estrogen receptor-alpha gene expression and activity via the ErbB2/PI 3-K/Akt pathway. PMID- 15759011 TI - The Major Themes from the Plenary Panel Session of the International Society of Exposure Analysis -- 2004 Annual Meeting on: The Application of Exposure Assessment to Environmental Health Science and Public Policy -- What has been Accomplished and What Needs to Happen before Our 25th Anniversary in 2014. PMID- 15759012 TI - Association analysis of the 1858C>T polymorphism in the PTPN22 gene in juvenile idiopathic arthritis and other autoimmune diseases. AB - A functional single nucleotide polymorphism, 1858C>T, in the PTPN22 gene, encoding a tyrosine phosphatase, has been reported to be associated with type I diabetes and some other autoimmune diseases. To further investigate whether this polymorphism may be a general susceptibility factor for autoimmunity, we performed an association study in five different autoimmune diseases, three previously not tested. We found an association with juvenile idiopathic arthritis (OR=1.41; P=0.04), not previously reported, and a tendency for an association with coeliac disease (OR=1.35; P=0.08). In primary sclerosing cholangitis, no association was observed (OR=0.95; P=0.8). Furthermore, we confirmed the increased risk in rheumatoid arthritis (OR=1.58; P=0.001), but could not find support for an association with systemic lupus erythematosus (OR=0.94; P=0.8). Altogether, we have provided further evidence of an association between autoimmune diseases and the 1858C>T polymorphism in PTPN22. PMID- 15759014 TI - Systemic delivery: the last hurdle? PMID- 15759013 TI - CUL1, a component of E3 ubiquitin ligase, alters lymphocyte signal transduction with possible effect on rheumatoid arthritis. AB - Ubiquitination affects various immune processes and E3 ubiquitin ligases (E3) play an important role in determining substrate specificity. We identified 11 human E3 ligase genes of potential importance in pathogenesis of autoimmune diseases by search of public databases and screened them for candidacy of biological investigation with case-control linkage disequilibrium tests on multiple SNPs in the genes using rheumatoid arthritis (RA) as a model of autoimmune diseases. Significant association with RA was observed in an SNP in intron 3 of Cullin 1 (CUL1) that affected transcriptional efficiency of the promoter activity in lymphocytic cell lines. Quantitative expression analysis revealed that CUL1 mRNA was highly detected in lymphoid tissues including spleen and tonsil, and was specifically expressed in T and B lymphocytes in fractionated peripheral leukocytes. Histological evaluation of tonsils indicated that CUL1 protein expression was relatively specific for maturing germinal centers. Suppression of CUL1 expression had influence on the phenotype of T-cell line, that is, it inhibited IL-8 induction, which is known to play an important role in the migration of inflammatory cells into the affected area seen in RA. Our data suggest that the regulation of CUL1 expression in immunological tissues may affect the susceptibility of RA via altering lymphocyte signal transduction. PMID- 15759015 TI - Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin. AB - Gene therapy for Duchenne muscular dystrophy (DMD) will require sustained expression of therapeutic dystrophins in striated muscles. Lentiviral vectors have a relatively large transgene carrying capacity and can integrate into nondividing cells. We therefore explored the use of lentiviral vectors for transferring genes into mouse skeletal muscle cells. These vectors successfully transferred a minidystrophin expression cassette into mdx muscles, and minidystrophin expression persisted and prevented subsequent muscle fiber degeneration for at least 6 months. However, only low to moderate levels of skeletal muscle transduction could be obtained by intramuscular injection of the highest currently available lentiviral doses. Using cultured cells, the lentiviral vectors effectively transduced proliferating and terminally differentiated muscle cells, indicating that cell cycling is not essential for transduction of myogenic cells. We further showed that lentiviral vectors efficiently transduced both primary myoblasts and multipotent adult progenitor cells (MAPCs) in vitro, and the cells persistently expressed transgenes without any obvious toxicity. When mdx primary myoblasts were genetically modified with minidystrophin vectors and transplanted into mdx skeletal muscles, significant numbers of dystrophin-expressing myofibers formed. Finally, we showed that a short, highly active CK6 regulatory cassette directed muscle-specific activity in the context of the lentiviral vectors. The ability of lentiviral vectors to transduce myogenic progenitors using a minidystrophin cassette regulated by a muscle-specific promoter suggests that this system could be useful for ex vivo gene therapy of muscular dystrophy. PMID- 15759016 TI - Polymers for cancer gene therapy: browsing the library. PMID- 15759018 TI - Gene transfer using the mature form of VEGF-D reduces neointimal thickening through nitric oxide-dependent mechanism. AB - Gene transfer to the vessel wall using vascular endothelial growth factors (VEGFs) has shown therapeutic potential for the treatment of restenosis. In this study, we evaluated the effect of catheter-mediated adenoviral (Ad) gene transfer of the mature form of VEGF-D (VEGF-D(DeltaNDeltaC)) in balloon-denuded cholesterol-fed rabbit aorta. AdLacZ was used as a control. Transduced VEGF D(DeltaNDeltaC) mRNA was detectable in the arterial wall with RT-PCR at 6, 14 and 28 days. Gene transfer efficiency as detected with X-gal staining 6 days after the AdLacZ transduction was 1.91 +/- 1.32% in intima. AdVEGF-D(DeltaNDeltaC) gene transfer led to 52% reduction in intima/media ratio (I/M) as compared to the AdLacZ controls at 14 days time point. At 6 days there were no differences in I/M, but the number of macrophages in the vessel wall was 85% lower in the AdVEGF D(DeltaNDeltaC) group as compared to the controls. The therapeutic effect was no longer detectable 28 days after the gene transfer. The therapeutic effect of VEGF D(DeltaNDeltaC) was nitric oxide (NO)-dependent as the feeding of NO synthase inhibitor, L-NAME, blocked the reduction in intimal thickening. It is concluded that AdVEGF-D(DeltaNDeltaC) gene transfer reduces intimal thickening and macrophage influx into the vessel wall in balloon-denuded rabbit aortas. PMID- 15759019 TI - Enhancement of adenoviral gene transfer to adult rat cardiomyocytes in vivo by immobilization and ultrasound treatment of the heart. AB - Direct injection of adenoviral vectors into ventricular myocardium in vivo produces local transfection of cells including cardiomyocytes. The use of vectors coexpressing GFP with the gene of interest allows subsequent identification of transfected myocytes isolated from the heart some days later, and examination of their function in cell bath experiments. We have injected vectors for antisense to phospholamban, or a control virus for expression of GFP only, into adult rat heart in vivo and then removed the heart and isolated ventricular myocytes 7 days later. Brief immobilization of the ventricle during and after injection using a haemoclip increased the number of transfected rod-shaped, viable myocytes from 1.7 +/- 0.8% (n = 8) to 5.6 +/- 0.8% (n = 9). This was further increased to 13.2 +/- 1.1% (n = 8) by the application of ultrasound pulses to the site before and after injection. Phospholamban antisense increased contraction amplitude and accelerated myocyte relengthening or decline of the Ca(2+) transient in transfected myocytes, while GFP control did not. Qualitative and quantitative effects of phospholamban downregulation were comparable between in vivo and in vitro transfections. This technique will have a number of uses, including production of transfected myocytes without the problem of culture-induced changes in contractility. PMID- 15759020 TI - Leptin is independently associated with systolic blood pressure, pulse pressure and arterial compliance in hypertensive African women with increased adiposity: the POWIRS study. AB - High leptin levels are often observed in human obesity and are implicated in obesity-related hypertension. Leptin levels have been found to be higher in hypertensive obese African-American women compared to normotensive African American women, but a direct association between leptin and blood pressure could not be obtained. Additionally, increased adiposity has been associated with higher aortic stiffness in obese African-American women, but leptin was not included in the study. The effects of leptin on cardiovascular function in African women have not yet been determined. We hypothesised that leptin is directly associated with blood pressure and decreased arterial compliance and that leptin levels are significantly higher in hypertensive overweight/obese African women compared to normotensive overweight/obese African women. A case case control study was performed which included 98 African women. The subjects were divided into lean normotensive (lean NT), overweight/obese normotensive (OW/OB NT) and overweight/obese hypertensive (OW/OB HT). The Finometer apparatus was used to obtain a more elaborate cardiovascular profile. Serum leptin and insulin levels as well as the HOMA-IR index were determined. Various anthropometric measures were obtained. Leptin levels were elevated (P < or = 0.05) in the OW/OB NT and HT groups compared to the lean NT group, but were similar in the OW/OB NT and HT groups. After adjusting for obesity, insulin resistance, hyperinsulinaemia and age, a direct positive correlation was obtained between leptin and systolic blood pressure (SBP) (P < or = 0.05; r = 0.68) in the OW/OB HT group. Additionally, leptin also correlated negatively with arterial compliance (P< or = 0.05; r = -0.76) and positively with pulse pressure (P < or = 0.05; r = 0.71) in the OW/OB HT group. In conclusion, even though leptin levels were the same in OW/OB HT and NT African women, leptin was directly and positively associated with SBP and pulse pressure and negatively with C(W) only in OW/OB HT African women, independent of obesity, insulin-resistance, hyperinsulinaemia and age. PMID- 15759021 TI - Reduction of blood pressure with calcium and potassium supplementation in children with salt sensitivity: a 2-year double-blinded placebo-controlled trial. AB - An investigation of the reductive effect of blood pressure and increment of urinary sodium excretion with calcium and potassium supplementation in children with sodium sensitivity is conducted. In total of 261 school children who had completed a 2-year double-blinded, placebo-controlled trial with calcium and potassium supplementation salt sensitivity, with a salt volume expansion and contraction protocol, was determined. The results showed that in children with salt sensitivity, the increase in blood pressure in the supplementary group was lower by 4.3/4.8 mmHg than that in the placebo group (P<0.05), while no significant change was found between the supplementary group and placebo group in children with nonsalt sensitivity. With calcium and potassium supplementation, the night urinary sodium excretion in children with salt sensitivity was significantly increased (P<0.01), and it is negatively correlated with the increase in blood pressure. It was suggested that a moderate increase of calcium and potassium intake in children with salt sensitivity, through interaction with sodium, can promote urinary sodium excretion and may play contribute to the prevention of hypertension. PMID- 15759022 TI - The 1964 trial of antihypertensive treatment. PMID- 15759023 TI - The effect of treatment on health-related quality of life in patients with hypertension and renal artery stenosis. AB - The quality of life in patients with hypertension is considered to be impaired mainly by side effects of antihypertensive drug therapy. Since balloon angioplasty for renal artery stenosis has a medication-sparing effect, it may lead to an improvement in quality of life. The objective of the study is to compare the effect of antihypertensive drug therapy and balloon angioplasty on quality of life in patients with hypertension and renal artery stenosis. We compared the quality of life in 56 patients randomised to balloon angioplasty to that in 50 patients randomised to antihypertensive drug therapy after 3 and 12 months of follow-up. Quality of life was measured using a questionnaire on physical symptoms associated with hypertension and antihypertensive drugs, and two generic health questionnaires (MOS Survey and EuroQol instrument). After follow-up, the patients who underwent angioplasty used less antihypertensive drugs than the patients who were treated with antihypertensive drugs only (mean+/ s.d., 1.9+/-0.9 vs 2.5+/-1.0 drugs after 3 months, P=0.002). They reported similar physical complaints, however, and a similar quality of life. The results after 12 months of follow-up were the same. In conclusion, for patients with hypertension and renal artery stenosis, the decrease in antihypertensive medication after intervention is too small to lead to a detectable improvement in quality of life. PMID- 15759024 TI - Lowering of blood pressure leads to decreased circulating interleukin-6 in hypertensive subjects. AB - Interleukin-6 (IL-6), the major proinflammatory cytokine, has been described to be associated with the hypertensive and atherosclerotic states. We aimed to explore whether the concentration of circulating IL-6 and adhesion molecules could be modified by decreasing blood pressure in hypertensive subjects. A total of 30 subjects (18 men), aged 34-48 years, were enrolled in this study, 17 hypertensive never-treated patients (HTA) and 13 normotensive subjects (C). HTA subjects were treated with irbesartan, 150-300 mg/day for 3 months, and serum IL 6, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, sP selectin, sE-selectin and monocyte chemoattractant protein-1 were measured at 0 and 12 weeks. The two study groups were similar in age, body mass index (BMI) and gender. At baseline, circulating IL-6 levels, but not adhesion molecules, were significantly associated with systolic blood pressure (r=0.41; P=0.03) and BMI (r=0.53; P=0.005). Systolic and diastolic blood pressure decreased significantly (P<0.01) in parallel to serum IL-6 levels (from 3.72+/-0.82 to 3.23+/-0.19 pg/ml, P=0.02) reaching a similar concentration to normotensive patients (3.33+/-0.3 pg/ml) after treatment with irbesartan. No significant changes were observed in any other of the tested parameters. In conclusion, the treatment of high blood pressure lowers circulating IL-6 in young hypertensive patients. PMID- 15759025 TI - High-dose monotherapy vs low-dose combination therapy of calcium channel blockers and angiotensin receptor blockers in mild to moderate hypertension. AB - The objectives of the study were to compare long-acting dihydropyridine calcium channel blockers (CCBs) with angiotensin II receptor blockers (ARBs) according to the ambulatory blood pressure monitoring (ABPM) profile in stage 1 and 2 newly diagnosed hypertensives and also to evaluate the efficacy of high-dose monotherapy vs low-dose combination therapy of the two drug categories among the subjects with inadequate blood pressure (BP) control after conventional low-dose monotherapy. We obtained 24-h ABPM readings from 302 subjects with newly diagnosed stage 1 or 2 essential hypertension. The study protocol consisted of initial drug treatment with a low dose of either CCBs or ARBs. Hypertensives who did not achieve BP control were randomized to high-dose monotherapy of either category of drug or low-dose combination therapy. CCBs and ARBs in low-dose monotherapy achieved BP control in 53.8 and 55.3% of the cases, respectively. However, subjects under treatment with CCBs experienced side effects more often and required that treatment be discontinued. Hypertensives who failed to control their BP with low-dose monotherapy did significantly better with low-dose combination treatment (61.6%) than with high-dose CCBs (42.8%) or ARBs (40.5%) monotherapy (P<0.05). In terms of ABPM, low-dose combination therapy exhibited better 24-h BP profile according to trough-to-peak ratio, hypertensive burden and BP variability. In conclusion, low-dose ARBs and CCBs have a comparable effect in subjects with grade 1 and 2 arterial hypertension. In hypertensives who are not controlled by low-dose monotherapy, low-dose combination therapy proves be more efficacious than high-dose monotherapy. PMID- 15759026 TI - Acute vs chronic volume overload on arterial stiffness in haemodialysis patients. PMID- 15759027 TI - Stopping oral contraceptives: an effective blood pressure-lowering intervention in women with hypertension. AB - The association of combined oral contraceptives (OC) with higher blood pressure (BP) may be attenuated with pills with smaller doses of oestrogen. The effect of stopping OC on BP of patients with hypertension was not described to date. In a cohort study of patients with hypertension, we identified 72 women using OC among 2112 patients seen from 1989 to 2002. Stopping hormonal contraception was recommended to all. The main outcome measurement was BP change in women who stopped (n=44) and who did not stop (n=28) OC, adjusting for baseline BP and age. Odds ratio for having a reduction of at least 20 mmHg in systolic blood pressure (SBP) or 10 mmHg in diastolic blood pressure (DBP), adjusting for age, change in weight and prescription of BP-lowering drugs, were calculated. The mean follow-up time was 6.6+/-7.5 months. Participants who stopped and did not stop OC had similar baseline characteristics. The deltas of SBP (adjusted) were 15.1+/-2.6 mmHg in patients who stopped and 2.8+/-3.2 mmHg in patients who did not stop OC (P=0.004). The corresponding values for DBP were 10.4+/-1.8 and 2.7+/-2.2 mmHg (P=0.008), respectively. The odds ratio (adjusted) for having a decrease of at least 20 mmHg in SBP or 10 mmHg in DBP was 0.28 (95% CI 0.08-0.90) in patients who stopped OC. Stopping OC is an effective antihypertensive intervention in a clinical setting. PMID- 15759028 TI - NSAID use and BP in treated hypertensives: a retrospective controlled observational study. AB - The objective of this study was to investigate the association between NSAID use and blood pressure (BP) among a sample of treated hypertensive patients. A controlled observational study was designed in UK primary care setting. Patients with diagnosed hypertension and currently being prescribed antihypertensive medication registered with four general practices, comparing patients also prescribed NSAIDs (exposed) to those not prescribed NSAIDs (unexposed). Majority of the patients were elderly. Systolic and diastolic pressure were the outcome measures. Data were collected for 184 NSAID users and 762 nonusers with a mean age of 68 years. There was no difference in either systolic (adjusted difference 1.9 mmHg, 95% CI -0.7 to 4.5, P=0.15) or diastolic (adjusted difference 1.0 mmHg, 95% CI -0.3 to 2.3, P=0.15) blood pressure. There was no evidence of any interactions according to categories of age, sex, or number of antihypertensive drugs prescribed. Among NSAID users, there was no evidence of any differences in blood pressure according to NSAID type or dose. In conclusion we found no evidence for an association between NSAID usage and BP control in known hypertensive patients receiving antihypertensive medication in primary care. The reported association between NSAID use and BP control appears much less substantial than has been previously suggested. PMID- 15759029 TI - Caspase- and mitochondrial dysfunction-dependent mechanisms of lysosomal leakage and cathepsin B activation in DNA damage-induced apoptosis. AB - A lysosomal pathway, characterized by partial rupture of lysosomal membranes and cathepsin B activation, is activated during camptothecin (CPT)-induced apoptosis in U937 and Namalwa cancer cells. These lysosomal events occur simultaneously with mitochondrial permeabilization and caspase activation. In U937 cells, blocking mitochondrial permeability transition pore with cyclosporin A and bongkrekic acid reduces mitochondrial and lysosomal rupture, suggesting that lysosomal rupture may be dependent, in part, on mitochondrial disruption. Overexpressing bcl-xL, an antiapoptotic protein known to preserve mitochondrial functions, also impedes lysosomal and mitochondrial disruption in both cell lines, indicating signaling between the two organelles. In addition, no evidence was obtained of bcl-2-like proteins targeting lysosomes. Caspase activities, including caspase-2L, are required for lysosomal and mitochondrial disruption, and lysosomal cathepsin B slightly participates in apoptosis propagation after CPT, although not essential for apoptosis activation. Our study provides evidence for the participation of a lysosomal pathway during DNA damage-induced cell death. Our data suggest that caspase activation and mitochondrial disruption represent cell-context-specific mechanisms by which DNA damage leads to lysosomal rupture, and that lysosomal cathepsins could slightly participate in apoptosis propagation after CPT. PMID- 15759030 TI - Mcl-1 gene promoter insertions do not correlate with disease outcome, stage or VH gene mutation status in chronic lymphocytic leukaemia. PMID- 15759032 TI - Transfer of genomics information to flow cytometry: expression of CD27 and CD44 discriminates subtypes of acute lymphoblastic leukemia. PMID- 15759031 TI - Combined analysis of ZAP-70 and CD38 expression as a predictor of disease progression in B-cell chronic lymphocytic leukemia. AB - Prognostic predictions in B-cell chronic lymphocytic leukemia (B-CLL) at early clinical stage are based on biological disease parameters, such as ZAP-70 and CD38 protein levels, genomic aberrations as well as immunoglobulin variable heavy chain gene (IgV(H)) mutation status. In the current study, ZAP-70 and CD38 expressions were examined by flow cytometry in 252 patients with B-CLL. Cytoplasmic ZAP-70 expression in more than 20% (ZAP-70(+)) and surface CD38 expression on more than 30% (CD38(+)) of B-CLL cells were associated with an unfavorable clinical course. The levels of ZAP-70 and CD38 did not change over time in the majority of patients where sequential samples were available for analysis. Combined analysis of ZAP-70 and CD38 yielded discordant results in 73 patients (29.0%), whereas 120 patients (47.6%) were concordantly negative and 59 patients (23.4%) were concordantly positive for ZAP-70 and CD38 expression. Median treatment-free survival times in patients whose leukemic cells were ZAP 70(+)CD38(+) was 30 months as compared to 130 months in patients with a ZAP-70( )CD38(-) status. In patients with discordant ZAP-70/CD38 results, the median treatment-free survival time was 43 months. Thus, ZAP-70 and CD38 expression analyses provided complementary prognostic information identifying three patient subgroups with good, intermediate and poor prognosis. Over-representation of high risk genomic aberrations such as 17p deletion or 11q deletion and distribution of the IgV(H) mutation status in B-CLL discordant for ZAP-70/CD38 pointed toward a distinct biologic background of the observed disease subgroups. This finding was also supported by microarray-based gene expression profiling in a subset of 35 patients. The expression of 37 genes differed significantly between the three groups defined by their expression of ZAP-70 and CD38, including genes that are involved in regulation of cell survival and chemotherapy resistance. PMID- 15759033 TI - Outcome of myeloablative allogeneic stem cell transplantation in multiple myeloma with a 153Sm-EDTMP-based preparative regimen. PMID- 15759034 TI - Good prognosis cytogenetics in B-cell chronic lymphocytic leukemia is associated in vitro with low susceptibility to apoptosis and enhanced immunogenicity. AB - Chromosomal abnormalities in B-cell chronic lymphocytic leukemia (B-CLL) have been shown to correlate with prognosis. Little is known about the relationship between chromosomal abnormalities and biological behavior of B-CLL cells in vitro. The present study was designed to explore the impact of chromosomal abnormalities determined by interphase fluorescence in situ hybridization (FISH) on the in vitro survival and immunogenicity of B-CLL. Considerable heterogeneity was noted in the in vitro survival and expression of costimulatory, adhesion, and antigen-presenting molecules by B-CLL cells. Spontaneous apoptosis of B-CLL cells in vitro was significantly lower in samples with good prognosis cytogenetics when compared to samples with poor prognosis cytogenetics. In contrast, B-CLL cells from samples with good prognosis cytogenetics exhibited higher basal expression of molecules involved in costimulation, cellular adhesion, and antigen presentation, and induced significantly more T-cell proliferation in mixed lymphocyte cultures. We conclude that chromosomal aberrations of B-CLL cells correlate with the in vitro biological behavior of B-CLL. Our data indicate that good prognosis cytogenetics correlates with less spontaneous apoptosis but greater in vitro immunogenicity. These findings could have significant implications on the design of future therapeutic approaches in patients with CLL, and the likelihood of response based on cytogenetics. PMID- 15759035 TI - Clinical and cytogenetic features of 508 Chinese patients with myelodysplastic syndrome and comparison with those in Western countries. AB - Myelodysplastic syndrome (MDS) is a clonal hematopoietic stem cell disorder characterized by ineffective hematopoiesis and leukemia progression. Racial differences may exist on clinical pictures and the molecular events leading to MDS, which are heterogeneous. To better define the clinical and cytogenetic features in Chinese patients, a retrospective multicentric study was performed in 508 MDS cases. Compared with Western countries, Chinese patients showed younger age (median: 49 vs 65-73 years), lower percentages of RARS (2.8 vs 6.6-15.3%), and CMML (5.2 vs 11.7-30.6%). Cytogenetically, among 367 cases with evaluable data, abnormal karyotypes were found in 136 cases, including 56 numerical and 80 structural changes. Incidences of single chromosome 5 and 7 abnormalities were lower than those in Western countries (2.2 vs 17.8-42.5%). However, complex cytogenetic aberrations and chromosome translocations were frequently observed and related to poor prognosis. Both multiple chromosome deletions and translocations were detected in advanced subtypes (RAEB and RAEB-T). Analysis of 200 cases revealed a higher incidence of hepatitis-B-virus infection than that in non-MDS population (21.00 vs 9.75%). This study further confirmed: (1) different genetic/environmental backgrounds between Asian and Western MDS populations; (2) a strong predictive value of cytogenetic abnormalities on disease outcome and involvement of genomic instability in leukemia clone development. PMID- 15759036 TI - Regulation of glucocorticoid receptor steroid binding and trafficking by the hsp90/hsp70-based chaperone machinery: implications for clinical intervention. PMID- 15759037 TI - mRNA expression levels of (co)chaperone molecules of the glucocorticoid receptor are not involved in glucocorticoid resistance in pediatric ALL. AB - Resistance to glucocorticoids (GC) is an important adverse risk factor in the treatment of acute lymphoblastic leukemia (ALL). To induce apoptosis, GC bind to the GC receptor (GR), which is regulated by various (co)chaperone proteins such as heat-shock protein 70 (HSP-70), HSP-40, HIP (HSP-70-interacting protein), BAG 1 (BCL-2-associated gene product-1), HOP (HSP-70/HSP-90-Organizing protein), HSP 90, P-23, FKBP-51, FKBP-52 and CYP-40. In this study, we tested the hypothesis that mRNA expression levels of these molecules are determinants of GC resistance in childhood ALL. In all, 20 children with ALL cells in vitro sensitive to prednisolone (LC(50) < 0.1 microg/ml) were compared each with a resistant patient (LC(50) >150 mug/ml), matched for immunophenotype, age and white blood cell count. mRNA expression levels of the (co)chaperone molecules were measured by quantitative real-time RT-PCR and normalized to GAPDH and RNaseP levels. In vitro resistance to prednisolone was measured by MTT assay. HSP-90 mRNA expression levels were 2000-fold higher as compared to HSP-70. Using matched pair analysis, mRNA expression levels of the various (co)chaperone molecules were not significantly different between in vitro-sensitive and -resistant patients. GC resistance in childhood ALL cannot be attributed to different mRNA expression levels of the investigated (co)chaperone molecules involved in GC binding and transport to the nucleus. PMID- 15759038 TI - Differential gene expression in hematopoietic progenitors from paroxysmal nocturnal hemoglobinuria patients reveals an apoptosis/immune response in 'normal' phenotype cells. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired stem cell disorder characterized clinically by intravascular hemolysis, venous thrombosis, and bone marrow failure. Despite elucidation of the biochemical and molecular defects in PNH, the pathophysiology of clonal expansion of glycosylphosphatidylinositol anchored protein (GPI-AP)-deficient cells remains unexplained. In pursuit of evidence of differences between GPI-AP-normal and -deficient CD34 cells, we determined gene expression profiles of isolated marrow CD34 cells of each phenotype from PNH patients and healthy donors, using DNA microarrays. Pooled and individual patient samples revealed consistent gene expression patterns relative to normal controls. GPI-AP-normal cells from PNH patients showed upregulation of genes involved in apoptosis and the immune response. Conversely, genes associated with antiapoptotic function and hematopoietic cell proliferation and differentiation were downregulated in these cells. In contrast, the PNH clone of GPI-AP-deficient cells appeared more similar to CD34 cells of healthy individuals. Gene chip data were confirmed by other methods. Similar gene expression patterns were present in PNH that was predominantly hemolytic as in PNH associated with aplastic anemia. Our results implicate an environmental influence on hematopoietic cell proliferation, in which the PNH clone evades immune attack and destruction, while normal cells suffer a stress response followed by programmed cell death. PMID- 15759039 TI - Animal models of acute myelogenous leukaemia - development, application and future perspectives. AB - From the early inception of the transplant models through to contemporary genetic and xenograft models, evolution of murine leukaemic model systems have been critical to our general comprehension and treatment of cancer, and, more specifically, disease states such as acute myelogenous leukaemia (AML). However, even with modern advances in therapeutics and molecular diagnostics, the majority of AML patients die from their disease. Thus, in the absence of definitive in vitro models which precisely recapitulate the in vivo setting of human AMLs and failure of significant numbers of new drugs late in clinical trials, it is essential that murine AML models are developed to exploit more specific, targeted therapeutics. While various model systems are described and discussed in the literature from initial transplant models such as BNML and spontaneous murine leukaemia virus models, to the more definitive genetic and clinically significant NOD/SCID xenograft models, there exists no single compendium which directly assesses, reviews or compares the relevance of these models. Thus, the function of this article is to provide clinicians and experimentalists a chronological, comprehensive appraisal of all AML model systems, critical discussion on the elucidation of their roles in our understanding of AML and consideration to their efficacy in the development of AML chemotherapeutics. PMID- 15759040 TI - Opinion: Cell entry machines: a common theme in nature? AB - Molecular machines orchestrate the translocation and entry of pathogens through host cell membranes, in addition to the uptake and release of molecules during endocytosis and exocytosis. Viral cell entry requires a family of glycoproteins, and the structural organization and function of these viral glycoproteins are similar to the SNARE proteins, which are known to be involved in intracellular vesicle fusion, endocytosis and exocytosis. Here, we propose that a family of bacterial membrane proteins that are responsible for cell-mediated adherence and entry resembles the structural architecture of both viral fusion proteins and eukaryotic SNAREs and might therefore share similar, but distinct, mechanisms of cell membrane translocation. Furthermore, we propose that the recurrence of these molecular machines across species indicates that these architectural motifs were evolutionarily selected because they provided the best solution to ensure the survival of pathogens within a particular environment. PMID- 15759041 TI - Biological control of soil-borne pathogens by fluorescent pseudomonads. AB - Particular bacterial strains in certain natural environments prevent infectious diseases of plant roots. How these bacteria achieve this protection from pathogenic fungi has been analysed in detail in biocontrol strains of fluorescent pseudomonads. During root colonization, these bacteria produce antifungal antibiotics, elicit induced systemic resistance in the host plant or interfere specifically with fungal pathogenicity factors. Before engaging in these activities, biocontrol bacteria go through several regulatory processes at the transcriptional and post-transcriptional levels. PMID- 15759042 TI - Regulation and biosynthesis of carbapenem antibiotics in bacteria. AB - Carbapenem antibiotics are members of the beta-lactam family of antibiotics, the most important class of antibiotics currently in clinical use. They are active against many important Gram-positive and Gram-negative pathogens. One important feature of carbapenem antibiotics is their resistance to several beta-lactamases. Thienamycin, isolated from Streptomyces cattleya, was the first carbapenem described. Other well-studied carbapenems were isolated from the Gram-negative bacteria Erwinia carotovora subsp. carotovora, Serratia sp. strain ATCC39006 and Photorhabdus luminescens strain TT01. Here, we review the genetics and biochemistry of carbapenem production in these bacteria. Research into carbapenems could uncover a new repertoire of bioactive molecules and biosynthetic enzymes, and exploiting these novel enzymes could lead to development of new classes of antibiotics with useful chemotherapeutic activities. PMID- 15759043 TI - Helicobacter pylori VacA, a paradigm for toxin multifunctionality. AB - Bacterial protein toxins alter eukaryotic cellular processes and enable bacteria to successfully colonize their hosts. In recent years, there has been increased recognition that many bacterial toxins are multifunctional proteins that can have pleiotropic effects on mammalian cells and tissues. In this review, we examine a multifunctional toxin (VacA) that is produced by the bacterium Helicobacter pylori. The actions of H. pylori VacA represent a paradigm for how bacterial secreted toxins contribute to colonization and virulence in multiple ways. PMID- 15759044 TI - Endothelial cell regulation of matrix metalloproteinases. AB - The process of sprouting angiogenesis requires that the endothelial cells degrade the basement membrane matrix and migrate into the interstitial matrix. Matrix metalloproteinases are enzymes capable of cleaving numerous extracellular matrix proteins. Increased production and activity of matrix metalloproteinases in any cell type is associated with a more migratory and invasive phenotype. This paper describes results of recent in-vitro studies of the regulation of transcription and activation of MMP-2 and MT1-MMP in endothelial cells, as well as studies that examined roles of matrix metalloproteinases in activity-induced angiogenesis. PMID- 15759045 TI - Chronic obstructive pulmonary disease: a novel risk factor for cardiovascular disease. AB - Chronic obstructive pulmonary disease (COPD) is one of the leading causes of morbidity and mortality in Canada and elsewhere. It affects 5% of all adult Canadians and is the fourth leading cause of death. Interestingly, the leading causes of hospitalizations and mortality among COPD patients are cardiovascular events. In the Lung Health Study, over 5 800 patients with mild to moderate COPD were studied. Forty-two to 48% of all hospitalizations that occurred over the study's 5-year follow-up period were related to cardiovascular complications. Various population-based studies suggest that independent of smoking, age, and gender, COPD increases the risk of cardiovascular morbidity and mortality twofold. Alarmingly, some bronchodilators, which are commonly used to treat symptoms in COPD, may increase the risk of cardiovascular morbidity and even mortality among COPD patients. In this paper, we discuss the epidemiologic evidence linking COPD and cardiovascular events as well as the potential mechanism(s) which may be responsible for this association. PMID- 15759046 TI - The role of notch in modeling and maintaining the vasculature. AB - The Notch proteins encompass a family of transmembrane receptors that have been highly conserved through evolution as mediators of cell fate, and are comprised of 4 members in mammals (Notch1 to Notch4). Following intra cellular processing of the full-length protein, Notch is expressed at the cell surface as a heterodimeric receptor. Engagement by ligand results in a 2-step cleavage of the Notch heterodimer, releasing the intracellular domain of Notch and allowing translocation to the nucleus. The intracellular domain of Notch interacts with the DNA-binding factor, CSL, resulting in transactivation at various promoters, in particular those of various basic helix-loop-helix factors of the HES (Hairy and Enhancer of Split) and HRT families (Hairy-Related Transcription factor). Recent findings implicate Notch as playing a critical and non-redundant role in vascular development and maintenance. This article briefly reviews vessel development and Notch signaling and highlights studies that examine Notch functions such as proliferation, cell survival, migration, adhesion, and mesenchymal transformation in the vasculature. Human diseases caused by Notch pathway members are also discussed. PMID- 15759047 TI - Activation of AMP-activated protein kinase (AMPK) inhibits protein synthesis: a potential strategy to prevent the development of cardiac hypertrophy. AB - A necessary mediator of cardiac myocyte enlargement is protein synthesis, which is controlled, in part, by the highly energy-consuming process of peptide-chain elongation. Recently, AMP-activated protein kinase (AMPK), which is a key regulator of cellular energy homeostasis, has been shown to phosphorylate a number of enzymes involved in the control of protein synthesis. Since AMPK may inhibit protein synthesis via a number of different pathways, it is possible that AMPK is also a key regulator of cardiac hypertrophy. Recent advances linking AMPK and the energy status of the cell to the regulation of protein synthesis and (or) cardiac myocyte hypertrophy will be discussed. PMID- 15759048 TI - Regulation of therapeutic apoptosis: a potential target in controlling hypertensive organ damage. AB - Cell growth and survival are potential therapeutic targets for the control of complications associated with hypertension. In most cardiovascular disorders, cardiac fibroblasts and large-vessel smooth muscle cells can replicate and thus contribute to the disease. We propose that cardiovascular hyperplasia may be reversed via therapeutic apoptosis induction with drugs that are safe and already used in the clinic. We first reported that, irrespective of the drug class, those drugs that are able to induce regression of cardiovascular hypertrophy are also able to reverse cardiovascular hyperplasia via apoptosis. Drugs active in this regard include inhibitors of the renin-angiotensin system, calcium channel blockers, and beta-blockers. Moreover, the effects of these drugs on cell survival is not merely secondary to blood pressure reduction. Therapeutic apoptosis in the cardiovascular system of the spontaneously hypertensive rat is characterized by a rapid and transient onset following initiation of antihypertensive treatment. Herein, the induction and termination of therapeutic apoptosis during drug treatment of hypertension will be briefly reviewed and supported by novel data suggesting that reversal of cardiovascular hyperplasia is associated with reduced cell growth and a resistance to further induction of therapeutic apoptosis, as shown in spontaneously hypertensive rats receiving an intermittent regime of nifedipine therapy. We propose that the presence of a cell subpopulation with defective cell cycle regulation may determine organ susceptibility to undergo therapeutic apoptosis. PMID- 15759049 TI - The value of studying gene-environment interactions in culturally diverse populations. AB - Cardiovascular disease (CVD) is the leading cause of death and disability in the world. It is anticipated that CVD will reach pandemic proportions by the year 2020. Although the major causes of CVD are well documented and explain the majority of cardiovascular deaths, the prevalence of conventional cardiovascular risk factors vary substantially across diverse cultural groups. These differences are attributed to cultural or genetic differences or to interactions between genes and environmental factors. Substantial efforts have been invested in determining the genetic influences on CVD development, and it is unlikely that a single gene is responsible for the development of atherosclerotic CVD or its classical risk factors such as blood pressure or plasma lipids. It is more plausible that multiple genes, acting either alone or in concert with one another, which display effect modification in the presence of certain environmental factors, are modestly associated with CVD or its main risk factors. Following this hypothesis, studying populations with diversity in environmental factors may increase the discovery potential of gene-environmental interactions. In this brief review, the advantage of studying gene-environment interactions across heterogeneous groups with diverse lifestyles is discussed. PMID- 15759050 TI - The role of endothelin-1 in myocarditis and inflammatory cardiomyopathy: old lessons and new insights. AB - Endothelin-1 has emerged as an important participant in the pathophysiology of a variety of cardiovascular diseases, where it may act on endocrine, paracrine and autocrine bases. Here we review its regulated biosynthesis, receptor-mediated signaling, and functional consequences in the heart, with particular emphasis on cardiac development and disease. Exploring published data employing molecular genetic mouse models of endothelin dysregulation, we highlight its heretofore underappreciated role as a pro-inflammatory cytokine. We also present novel micro array data from one such mouse model, which implicate the specific downstream pathways that may mediate endothelin-1's effects. PMID- 15759051 TI - The pro-oxidant role of methylglyoxal in mesenteric artery smooth muscle cells. AB - Methylglyoxal (MG), a highly reactive metabolite of glucose, causes non-enzymatic glycation of proteins to form irreversible advanced glycation endproducts (AGEs). The present study investigated whether methylglyoxal induced oxidative stress and activated nuclear factor kappa B (NF-kappaB) in freshly isolated and cultured smooth muscle cells (SMCs) from rat mesenteric artery. The treatment of cells with MG (50 or 100 micromol/L) induced a significant increase in AGE formation and oxidation of DCF. MG-enhanced generation of AGEs and the oxidation of DCF was markedly inhibited by antioxidant n-acetylcysteine (NAC, 600 micromol/L). MG at a concentration of 100 micromol/L increased the heme-oxygenase-1 expression in these cells. Moreover, MG activated NF-kappaB p65, indicated by an increased immuno cytochemistry stain for NF-kappaB p65 located in the nucleus after the treatment of mesenteric artery SMCs with MG. MG-induced activation of NF-kappaB p65 was inhibited by NAC. In summary, MG significantly increases oxidative stress and activates NF-kappaB p65 in mesenteric artery SMCs. The pro-oxidant role of methylglyoxal may contribute to various pathological changes of SMCs from resistance arteries. PMID- 15759052 TI - Neutrophil primary granule release and maximal superoxide generation depend on Rac2 in a common signalling pathway. AB - Neutrophils play an integral role in innate immunity by undergoing degranulation and respiratory burst in response to inflammatory stimuli. Rac2, a monomeric GTP binding protein, has been shown to be involved in several neutrophil functions, including primary granule release and superoxide (O(2)(-.) generation. We hypothesized that Rac2 is a common signalling molecule required for primary granule translocation and maximal O(2)(-.) production. Using bone marrow neutrophils from Rac2 knockout (KO) mice and wild type C57Bl/6 mice, we found that primary granule elastase and myeloperoxi dase release were absent in Rac2 KO neutrophils upon chemoattractant stimulation. Rac2 KO neutrophils also failed to produce maximal levels of extracellular O(2(-.) generation in response to phorbol myristate acetate (PMA). Although PMA was ineffective at eliciting primary granule mediator release, it induced secondary granule exocytosis in both WT and Rac2 KO neutrophils. Thus, the signalling pathway leading to primary granule release utilized Rac2, which was also necessary for full activation of O(2)(-.) generation in stimulated neutrophils. These findings indicate that O(2)(-.) release and secondary granule secretion may use protein kinase C (PKC) - dependent pathways, whereas primary granule exocytosis appears to rely on PKC independent signalling events. These findings shed light on possible signalling mechanisms involved in granule secretion from activated neutrophils responding to different stimuli. PMID- 15759053 TI - Neonatal maternal separation enhances dopamine D(2)-receptor and tyrosine hydroxylase mRNA expression levels in carotid body of rats. AB - Adult male (but not female) rats previously subjected to neonatal maternal separation (NMS) are hypertensive and show a significant increase (25%) in their hypoxic ventilatory response. To begin investigating the mechanisms involved in this gender-specific disruption in cardiorespiratory regulation, we tested the hypothesis that NMS alters the expression of dopamine D(2)-receptors and tyrosine hydroxylase mRNA in 3 peripheral organs involved in cardio respiratory regulation: the carotid bodies, superior cervical ganglia, and adrenals. Pups subjected to NMS were placed in a temperature- and humidity-controlled incubator 3 h per day for 10 consecutive days (P3-P12). Control pups were undisturbed. Once they reached adulthood (8-10 weeks), male and female rats were anesthetised. The carotid bodies, superior cervical ganglia, and adrenals were harvested for semi quantitative analyses of dopamine D(2)-receptors and tyrosine hydroxylase mRNA expression using reverse transcription-polymerase chain reaction (carotid bodies only) and Northern blot. In the carotid bodies, comparison of densitometric analyses showed that NMS enhanced tyrosine hydroxylase mRNA expression in male, but not female, rats. Neonatal maternal separation increased dopamine D(2) receptor mRNA expression also, but the effect was not gender specific. No changes in mRNA expression related to dopaminergic neurotransmission were observed in superior cervical ganglia or the adrenals. These results indicate that subsequent mechanistic investigations should focus on the carotid bodies, as enhancement of dopaminergic neurotransmission within this organ likely contributes to the gender specific effects of NMS on cardiorespiratory regulation. PMID- 15759054 TI - Effect of nonsteroidal anti-inflammatory drugs with varying extent of COX-2-COX-1 selectivity on urinary sodium and potassium excretion in the rat. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have different selectivity to inhibit cyclooxygenase-1 (COX-1) and COX-2. Treatment with NSAIDs has been associated with kidney side effects. We compared the effect of a selected group of NSAIDs with different COX-2--COX-1 selectivities on urinary sodium and potassium excretion in rats. Each treatment with rofecoxib, celecoxib, meloxicam, diclofenac, and flurbiprofen (30, 120, 9, 30, and 125 mg/kg, respectively) and placebo was administered orally once daily for 4 days. Urine was collected 0-8 h after each dose. Urinary sodium and potassium excretion and urine flow rate were compared with placebo. As compared with placebo, rofecoxib, celecoxib, diclofenac, and flurbiprofen significantly reduced excretion rate of sodium (rofecoxib, 0.28 +/- 0.02 vs. 0.41 +/- 0.03; celecoxib, 0.23 +/- 0.03 vs. 0.48 +/ 0.04; diclofenac, 0.09 +/- 0.02 vs. 0.46 +/- 0.03; and flurbiprofen, 0.11 +/- 0.02 vs. 0.47 +/- 0.02 micromol/(min x 100 g)) and potassium (rofecoxib, 0.55 +/- 0.04 vs. 0.68 +/- 0.04; celecoxib, 0.50 +/- 0.06 vs. 0.72 +/- 0.06; diclofenac, 0.26 +/- 0.05 vs. 0.67 +/- 0.04; and flurbiprofen, 0.35 +/- 0.05 vs. 0.62 +/- 0.03 micromol/ (min x 100 g)). Rofecoxib and flurbiprofen significantly reduced urine flow rate. Meloxicam had no significant effect on either sodium and potassium excretion or on the urine flow rate. At the examined dosage level, no relationship was found between reported COX-2--COX-1 selectivity and urinary electrolytes excretion. PMID- 15759055 TI - Role of the actin cytoskeleton in angiotensin II signaling in human vascular smooth muscle cells. AB - Angiotensin II (Ang II) regulates vascular smooth muscle cell (VSMC) function by activating signaling cascades that promote vasoconstriction, growth, and inflammation. Subcellular mechanisms coordinating these processes are unclear. In the present study, we questioned the role of the actin cytoskeleton in Ang II mediated signaling through mitogen-activated protein (MAP) kinases and reactive oxygen species (ROS) in VSMCs. Human VSMCs were studied. Cells were exposed to Ang II (10-7 mol/L) in the absence and presence of cytochalasin B (10-6 mol/L, 60 min), which disrupts the actin cytoskeleton. Phosphorylation of p38MAP kinase, JNK, and ERK1/2 was assessed by immuno blotting. ROS generation was measured using the fluoroprobe chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (4 micromol/L). Interaction between the cytoskeleton and NADPH oxidase was determined by evaluating the presence of p47phox in the Triton X-100 insoluble membrane fraction. Ang II significantly increased phosphorylation of p38MAP kinase, JNK, and ERK1/2 (two- to threefold above control, p < 0.05). Cytochalasin B pretreatment attenuated p38MAP kinase and JNK effects (p < 0.05) without altering ERK1/2 phosphorylation. ROS formation, which was increased in Ang II stimulated cells, was significantly reduced by cytochalasin B (p < 0.01). p47phox, critically involved in NADPH oxidase activation, colocalized with the actin cytoskeleton in Ang II stimulated cells. Our data demonstrate that Ang II mediated ROS formation and activation of p38MAP kinase and JNK, but not ERK1/2, involves the actin cytoskeleton in VSMCs. In addition, Ang II promotes interaction between actin and p47phox. These data indicate that the cytoskeleton is involved in differential MAP kinase signaling and ROS generation by Ang II in VSMCs. Together, these studies suggest that the cytoskeleton may be a central point of crosstalk in growth- and redox-signaling pathways by Ang II, which may be important in the regulation of VSMC function. PMID- 15759056 TI - Congestive heart failure complicating non-ST segment elevation acute coronary syndrome: incidence, predictors, and clinical outcomes. AB - There are limited data regarding the incidence and clinical significance of congestive heart failure (CHF) in patients with non-ST segment elevation acute coronary syndromes (ACS). The objectives of this study were to examine the incidence, predictors, and clinical outcomes in patients with ACS without ST elevation who develop CHF. We studied patients with unstable angina or non-ST segment elevation myocardial infarction (NSTEMI) randomized to hirudin or unfractionated heparin in the Organisation to Assess Strategies for Ischemic Syndromes (OASIS-2) trial. The diagnosis of CHF was based on a combination of clinical and radiographic features. Patients were followed for 6 months. Of 10 141 randomized patients, 501 (4.9%) developed CHF within the first week and 643 (6.3%) during 6 months of followup. Independent predictors for the development of CHF were older age, female sex, diabetes, prior MI, prior CHF, and NSTEMI at presentation. Compared with patients who did not develop CHF, patients who developed CHF were at increased risk of death (odds ratio (OR) 3.4, 95% CI 2.7 4.3), new MI (OR 2.8, 95% CI 2.2-3.6), and the need for intra-aortic balloon pump insertion (OR 5.4, 95% CI 3.5-8.4) at 7 days and 6 months. There was no increase in use of cardiac catheterization (OR 0.8, 95% CI 0.7-1.0) or revascularization (OR 0.9, 95% CI 0.7-1.1) in patients who developed CHF. CHF is a common complication in patients presenting with non-ST segment elevation ACS and is strongly associated with adverse clinical outcomes including new MI and death. Despite this worse prognosis, patients with ACS developing CHF are less likely to be referred for invasive management. PMID- 15759057 TI - Distribution of phenotypically disparate myocyte subpopulations in airway smooth muscle. AB - Phenotype and functional heterogeneity of airway smooth muscle (ASM) cells in vitro is well known, but there is limited understanding of these features in vivo. We tested whether ASM is composed of myocyte subsets differing in contractile phenotype marker expression. We used flow cytometry to compare smooth muscle myosin heavy chain (smMHC) and smooth muscle-alpha-actin (sm-alpha-actin) abundance in myocytes dispersed from canine trachealis. Based on immunofluorescent intensity and light scatter characteristics (forward and 90 degrees side scatter), 2 subgroups were identified and isolated. Immunoblotting confirmed smMHC and sm-alpha-actin were 10- and 5-fold greater, respectively, in large, elongate myocytes that comprised approximately 60% of total cells. Immunohistochemistry revealed similar phenotype heterogeneity in human bronchial smooth muscle. Canine tracheal myocyte subpopulations isolated by flow cytometry were used to seed primary subcultures. Proliferation of subcultures established with myocytes exhibiting low levels of smMHC and sm-alpha-actin was approximately 2 x faster than subcultures established with ASM cells with a high marker protein content. These studies demonstrate broad phenotypic heterogeneity of myocytes in normal ASM tissue that is maintained in cell culture, as demonstrated by divergent proliferative capacity. The distinct roles of these subgroups could be a key determinant of normal and pathological lung development and biology. PMID- 15759058 TI - Radiation-induced strain differences in mouse alveolar inflammatory cell apoptosis. AB - Whole-thorax irradiation results in the development of the diffuse inflammatory response alveolitis in C3H/HeJ (C3H) mice and a milder alveolitis with fibrosis in C57BL/6J (B6) mice. In this study, we investigate if this mouse strain difference in response to radiation is due to differences in lung inflammatory cell apoptosis. Mice of the C3H and B6 strains were given a radiation dose of 18 Gy to the thorax and the animals were sacrificed at 11 or 18 weeks following exposure or when they were moribund. Active caspase-3 staining was used to identify apoptotic cells in the alveolar space of histological lung sections from the mice. The apoptotic index of B6 mice was greater than that of C3H mice at 11 weeks postirradiation (17.8% of airspace cells vs. 7.8%, p = 0.028) and in mice sacrificed because of illness (27.3% vs. 14.4%, p = 0.036). No C3H mice survived to the later time point. The inflammatory cells undergoing apoptosis in the mouse lungs were morphologically consistent with alveolar macrophages. We conclude that a difference in inflammatory cell apoptosis may contribute to the disparate pulmonary radiation response of these mouse strains. PMID- 15759059 TI - Prostate cancer survivors' and partners' self-reports of health-related quality of life, treatment symptoms, and marital satisfaction 2.5-5.5 years after treatment. AB - PURPOSE/OBJECTIVES: To describe health-related quality of life (QOL), health status, and marital satisfaction of couples as many as 5.5 years after treatment for prostate cancer. DESIGN: Survey with longitudinal, comparative, and predictive elements. SETTING: A tertiary care nonprofit medical center in the southwestern United States. SAMPLE: Convenience sample of prostate cancer survivors (192 enrolled, 137 completed) and their partners (126 enrolled, 104 completed). Men averaged 70 years of age, women 66. Most men (86%) and women (89%) were white, and 71% had at least some college education. METHODS: Questionnaires were mailed annually. Women were enrolled 3.5 years after their partners were treated. Study participants received separate questionnaire packets. MAIN RESEARCH VARIABLES: Health-related QOL, health status including post-treatment symptoms, and marital satisfaction. FINDINGS: Men's health-related QOL, general physical health, and vitality decreased; urinary and sexual post treatment symptoms increased. Men were concerned about their sexual functioning although few sought treatment. Couples' health-related QOL and marital satisfaction were associated more closely than their health status. CONCLUSIONS: Regardless of type of treatment, health-related QOL and general health tend to decrease for prostate cancer survivors; men in watchful waiting tended to have poorer health outcomes. Men are concerned about sexual functioning, yet few are taking steps to remedy problems. Couples' health-related QOL and marital satisfaction are linked; however, health status indicators are less associated. IMPLICATIONS FOR NURSING: Nurses are in a key position to assess health-related QOL and sexual functioning concerns for prostate cancer survivors and their partners. PMID- 15759060 TI - Operationalizing symptom distress in adults with cancer: a literature synthesis. AB - PURPOSE/OBJECTIVES: To address inconsistencies in the definition and operationalization of symptom distress by synthesizing the literature on cancer related symptom distress in adults. DATA SOURCES: Electronic nursing, psychology, and medicine databases; online meeting abstracts; and various print sources. DATA SYNTHESIS: Eight distinct methods of operationalizing the concept were identified. Gender, ethnic, developmental, cultural, and individual differences in symptom distress have not been identified. Relationships among symptom frequency, intensity, and distress are unclear. CONCLUSIONS: Lack of clarity and consensus in defining and operationalizing symptom distress hinder meta-analysis, research synthesis, and research utilization. Symptom distress may be emerging as a component of the multidimensional symptom experience. IMPLICATIONS FOR NURSING: Defining and operationalizing symptom distress consistently will enhance research synthesis and assist clinicians with more effectively meeting the needs of people with cancer. Research is needed to identify the meanings of symptom distress to patients with cancer and to differentiate symptom distress from symptom frequency and intensity. PMID- 15759062 TI - The delay and worry experience of African American women with breast cancer. AB - PURPOSE/OBJECTIVES: To examine the delay in seeking treatment and worry experiences of African American women with breast cancer. DESIGN: Descriptive, correlational. SETTING: Urban northeastern United States. SAMPLE: 60 African American women diagnosed with breast cancer. METHODS: Consenting participants completed the worry subscale of the Ware Health Perception questionnaire and a Demographic and Illness-Related Information Sheet during a one-hour personal interview. Data analysis consisted of descriptive statistics and Pearson correlations. MAIN RESEARCH VARIABLES: Delay in seeking treatment, worry about breast cancer and symptoms, and sociodemographic characteristics. FINDINGS: Contrary to the literature, participants reported short patient and provider delay. As a result of little variability in delay, predicting those for whom worry was a deterrent or a motivator to seek prompt treatment was not possible. CONCLUSIONS: Although delay does exist, African American women with sociodemographic characteristics similar to white women who do not delay are likely to have similarly short symptom durations. Further study to determine who is helped and who is hurt by worry and other possible intervening factors would be useful. Including biologic characteristics such as tumor staging and hormone receptor information in future studies would allow for a closer examination of stage at diagnosis and biologic influence. IMPLICATIONS FOR NURSING: Interventions with African American women cannot assume that delay exists. Strategies that consider both individual and cultural group differences are essential to the early seeking of a diagnosis and treatment for breast cancer symptoms among African American women. PMID- 15759063 TI - Post-treatment symptoms among ethnic minority breast cancer survivors. AB - PURPOSE/OBJECTIVES: To determine racial and ethnic differences in the range and number of post-treatment symptoms among women who have undergone surgical and postsurgical treatment for breast cancer. DESIGN: Retrospective study. SETTING: Community-based social services agency and public health clinic. METHODS: Face-to face interview of 116 women who had undergone breast cancer surgery. SAMPLE: Primarily low-income women in an urban area in northern California. Participants were recruited via posted flyers regarding the study. MAIN RESEARCH VARIABLES: Outcomes (depression, fatigue, pain, and swelling from lymphedema) and demographics (ethnicity, age, income, insurance, education, and marital status). FINDINGS: The typical participant was 47 years old, had 12 years of education, and was a parent (74%), unmarried (67%), heterosexual (88%), uninsured (68%), and employed (66%). Thirty percent were Caucasian, 30% African American, 25% Latina, and 15% women of other ethnic backgrounds. African American women and Latinas reported increased rates of pain and an increased number of symptoms. Latinas also reported higher rates of fatigue and depression. In multivariate analyses, an increased number of symptoms were associated with decreased income, with receiving chemotherapy, with having a mastectomy, and with Latina ethnicity. CONCLUSIONS: These data suggest an increased rate of post-treatment symptoms experienced by low-income and ethnic minority women. IMPLICATIONS FOR NURSING: A need exists for affordable, culturally appropriate symptom management interventions. Nursing will have a vital role in designing, testing, and offering such interventions. PMID- 15759064 TI - Lessons learned in developing a support intervention for African American women with breast cancer. AB - PURPOSE/OBJECTIVES: To describe the process of planning a theory-based support group project for African American women with breast cancer. DATA SOURCES: A needs assessment and recruitment strategies consistent with the Oncology Nursing Society's Multicultural Outcomes: Guidelines for Cultural Competence were used to adapt a support group intervention for newly diagnosed African American women in urban central Texas. DATA SYNTHESIS: The reviewed literature and local cancer survivor leaders indicated the need for education and support of newly diagnosed women. Although researchers worked for several years with lay leaders to gain legitimacy and trust, not enough participants were recruited to test an intervention specifically for African American women. CONCLUSIONS: Recruiting support group research participants from a relatively small minority population is problematic even when collaborating with population leaders. IMPLICATIONS FOR NURSING: Nurses may encounter barriers to conducting research in minority populations. Starting early to build credibility with that population, being flexible with eligibility criteria, beginning with pre-experimental studies, and paying participants may be required. PMID- 15759065 TI - Telephone interpersonal counseling with women with breast cancer: symptom management and quality of life. AB - PURPOSE/OBJECTIVES: To examine the effectiveness of a telephone interpersonal counseling (TIP-C) intervention compared to a usual care attentional control for symptom management (depression and fatigue) and quality of life (positive and negative affect, stress) for women with breast cancer. DESIGN: Experimental with repeated measures. SETTING: Academic cancer center and urban, private oncology offices. SAMPLE: 48 women with breast cancer who were in their mid-50s, married, and employed at the time of the study. METHODS: Women were assigned to either the six-week TIP-C or attentional usual care groups. Women were matched on stage and treatment. Data were collected at baseline, after the six interventions, and one month postintervention. Measures included the Center for Epidemiologic Studies Depression Scale, Positive and Negative Affect Schedule, Multidimensional Fatigue Inventory, and Index of Clinical Stress. MAIN RESEARCH VARIABLES: Depression, positive and negative affect, fatigue, and stress. FINDINGS: Women in the intervention group experienced decreases in depression, fatigue, and stress over time and increases in positive affect. CONCLUSIONS: The preliminary results partially supported the effectiveness of TIP-C for symptom management and quality of life. The authors hypothesized that decreased depression, reduced negative affect, decreased stress, decreased fatigue, and increased positive affect over time would be the resulting psychosocial effects, given the theoretical underpinnings of the intervention. IMPLICATIONS FOR NURSING: Nurses need to assess the quantity and quality of the social support network early in treatment; women with less social support need to be referred to counseling and support services. Because these women have limited participation in face-to-face interventions, they should be encouraged to participate in telephone or online support programs or in other programs or organizations (e.g., churches, social clubs) that would provide support. PMID- 15759067 TI - The nurse as principal investigator in a pharmaceutically sponsored drug trial: considerations and challenges. AB - PURPOSE/OBJECTIVES: To discuss the process, considerations, benefits, and challenges of the nurse as principal investigator in a cancer care drug trial. DATA SOURCES: Published articles, anecdotal experience, and completed research studies. DATA SYNTHESIS: The specific processes that must be considered are funding sources, protocol development, trial implementation, dissemination of results, and ethical implications involved in industry sponsorship. Specific protocols are designed for evaluating adverse events. Working with pharmaceutical companies to receive financial support offers advantages but poses additional issues for consideration. CONCLUSIONS: Nurses can serve successfully as principal investigators in medication trials for cancer care. Regulatory bodies and specific procedures, as well as general considerations, mandate and guide investigator conduct when embarking on a pharmaceutical trial. IMPLICATIONS FOR NURSING: Oncology nurse researchers can look to pharmaceutical companies for potential funding in the evaluation of medications used in cancer care. PMID- 15759066 TI - Oncology Nursing Society year 2004 research priorities survey. AB - PURPOSE/OBJECTIVES: To determine the Oncology Nursing Society (ONS) research priorities for 2005-2008 for oncology nursing across the entire scope of cancer care, including prevention, detection, treatment, survivorship, and palliative care. DESIGN: Descriptive, cross-sectional survey. SAMPLE: Stratified into two groups: random sample of general membership (N = 2,205; responses = 287, or 13%) and all ONS active members in the United States with doctoral degrees (N = 627, responses = 144, or 23%); overall response rate was 15%. METHODS: The 2000 survey was revised and updated. Postcards were mailed to the original sample (N = 1,605) prior to the launch of the online survey, inviting participation via an online or paper-and-pencil survey. An e-mail announcement of the survey was launched one week later, followed by reminders the following week. Because of low response rates, a second sample (N = 600) was selected and contacted. MAIN RESEARCH VARIABLES: 117 topic questions divided into seven categories. Several items were new or reworded. FINDINGS: The top 20 research priorities included 12 of the top 20 items found in the 2000 survey; 8 topics were new to the top 20. Priority topics were distributed across six of seven categories. When general membership results were compared to the doctoral sample, 10 topics were among the top 20 for both groups. Nine topics were top priorities in the 2000 (researcher) and 2004 (doctorally prepared) surveys. CONCLUSIONS: Response rates to the electronic survey were lower than for previous paper-and-pencil surveys, but an adequate response was obtained. Rank order of mean importance ratings was determined by narrow differences in scores. The general membership and doctorally prepared samples showed similarities as well as differences in results. IMPLICATIONS FOR NURSING: The 2004 survey results will inform the 2005 research agenda and assist the ONS Foundation and other funding organizations in distributing research funds. PMID- 15759068 TI - Chemotherapy-induced peripheral neuropathy. AB - PURPOSE/OBJECTIVES: To review the literature documenting the scope, treatment, and prevention of chemotherapy-induced neuropathy. DATA SOURCES: Published abstracts, primary research literature, and textbook chapters. DATA SYNTHESIS: Recent improvements in the management of other treatment-related toxicities have led to peripheral neuropathy becoming a dose-limiting toxicity of commonly used chemotherapeutic groups such as platinols, vinca alkaloids, and taxanes. CONCLUSIONS: The nervous system has not been the focus of education or training for oncology nurses. Therefore, nurses' ability to educate patients regarding this aspect of their condition has been limited. IMPLICATIONS FOR NURSING: With its significant impact on quality of life, peripheral neuropathy treatment and prevention are important components in the care of patients with cancer. PMID- 15759069 TI - Oncology nurses' knowledge, attitudes, and experiences regarding advance directives. AB - PURPOSE/OBJECTIVES: To determine oncology nurses' knowledge of, attitudes toward, and experiences with advance directives (ADs). DESIGN: Descriptive, correlation survey. SAMPLE: Usable responses from 794 (21% return) of 3,840 randomly selected members of the Oncology Nursing Society. The typical respondent was female, Caucasian, married or living as married, middle-aged, and Christian. METHODS: A mailed survey using the Knowledge, Attitudinal, and Experiential Survey on Advance Directives instrument. MAIN RESEARCH VARIABLES: Knowledge, attitudes, experiences, confidence, and ADs. FINDINGS: Overall, oncology nurses were most knowledgeable about ADs in general (70% correct) and less knowledgeable about the Patient Self-Determination Act (51% correct) and their state laws (53% correct). The mean total knowledge score based on the three subscales was 17.4 out of a possible 30, or 58% correct. The nurses' experience with ADs was measured using a five-item subscale with a mean score of 4 (SD = 1.11). They were less confident in their ability to assist patients with completing ADs. Respondent attitudes reflected an advocacy role in end-of-life decisions. Attitude items were reviewed individually. Respondents strongly agreed (97%) with the statement that patients should receive sufficient medication to relieve pain even though it may hasten death, which reflects the emphasis in oncology on adequate pain management at the end of life. CONCLUSIONS: Nurses' knowledge scores were low. Nurses in the study were not highly confident in their ability to assist patients with ADs. Demographic variables generally did not influence knowledge, confidence, or experience scores. IMPLICATIONS FOR NURSING: More education related to ADs is needed and could be administered through in-service classes or continuing education. Nurses' responses indicated that they need more time to assist patients with completing ADs. This is difficult in the current practice environment but must be recognized as critically important. PMID- 15759071 TI - Aromatase inhibitor agents in breast cancer: evolving practices in hormonal therapy treatment. AB - PURPOSE/OBJECTIVES: To review the role of aromatase inhibitor agents with regard to current treatment strategies with hormonal therapy for women with breast cancer. DATA SOURCES: Published articles and books. DATA SYNTHESIS: Hormonal therapy is an essential component of the treatment of most women with breast cancer. Aromatase inhibitor agents are becoming an integral part of treatment for women with metastatic breast cancer and recently have become much more prominent in the treatment of women with early-stage breast cancer. The exact role of these agents in adjuvant therapy of breast cancer, either sequentially with the "gold standard" tamoxifen or for the duration of therapy, has yet to be determined. CONCLUSIONS: Recent studies with aromatase inhibitor agents are intriguing and suggest an improved side-effect profile and efficacy. The approval of these agents for the adjuvant treatment of breast cancer has led to a significant change in practice. IMPLICATIONS FOR NURSING: Breast cancer is an extremely common cancer in women, and oncology nurses take care of large numbers of patients with this disease. Oncology nurses need the most recent information so they can discuss aromatase inhibitor agents and therapy with their patients. PMID- 15759070 TI - Chemotherapy-induced cognitive impairment in women with breast cancer: a critique of the literature. AB - PURPOSE/OBJECTIVES: To review and critique the studies that have investigated chemotherapy-induced impairments in cognitive function in women with breast cancer. DATA SOURCES: Published research articles and textbooks. DATA SYNTHESIS: Although studies of breast cancer survivors have found chemotherapy-induced impairments in multiple domains of cognitive function, they are beset with conceptual and methodologic problems. Findings regarding cognitive deficits in women with breast cancer who currently are receiving chemotherapy are even less clear. CONCLUSIONS: Although data from published studies suggest that chemotherapy-induced impairments in cognitive function do occur in some women with breast cancer, differences in time since treatment, chemotherapy regimen, menopausal status, and neuropsychological tests used limit comparisons among the various studies. Further studies need to be done before definitive conclusions can be made. IMPLICATIONS FOR NURSING: The potential for chemotherapy-induced impairments in cognitive function may influence patients' ability to give informed consent, identify treatment toxicities, learn self-care measures, and perform self-care behaviors. PMID- 15759072 TI - Out of necessity: oncology nurses' experiences integrating the internet into practice. AB - PURPOSE/OBJECTIVES: To understand the experiences of oncology nurses who use the Internet in their practice when their patients use the Internet for cancer care. RESEARCH APPROACH: Heideggerian hermeneutics branch of phenomenology. SETTING: Oncology nurses were interviewed at their practice settings (n = 13), the researcher's office (n = 5), or their homes (n = 2). PARTICIPANTS: 20 nurses recruited from local and national Oncology Nursing Society meetings. Their practice sites were cancer centers, hospitals, clinics, veterans centers, communities, and Internet companies. METHODOLOGIC APPROACH: Data were collected by informal interviews that provided the narrative stories for hermeneutic analysis. MAIN RESEARCH VARIABLES: Internet use for cancer care, nurse-patient relationships, and Internet use for nursing practice. FINDINGS: Five related themes emerged: (a) varying degrees of Internet integration in the practice environment, (b) changing schools of thought, (c) developing Internet use for professional practice, (d) redefining relationships, and (e) new nursing skills. The two constitutive patterns are (a) integrating Internet into practice out of necessity and (b) reflecting historical changes in practice patterns influenced by technology. CONCLUSIONS: Nurses who use the Internet are developing new practice patterns that incorporate technology and foster nurse-patient partnerships. Practice environments either foster or hinder technology use. Nurse computer competency is essential. INTERPRETATION: Findings reflect the influence of adapting technology on practice. Internet use is a catalyst for redefining nurse-patient relationships into partnerships. The movement suggests a need for nurse Internet competencies, environmental support, and consideration for patient access (digital divide). Evaluation of content in nursing curricula and of patient competencies is advised. Further research on patient experiences is recommended. PMID- 15759073 TI - Do patients' beliefs act as barriers to effective pain management behaviors and outcomes in patients with cancer-related or noncancer-related pain? AB - PURPOSE/OBJECTIVES: To understand the role of patients' beliefs in pain management in a cancer population treated in a primary care setting. DESIGN: Secondary analyses of data from the baseline phase of a randomized study. SETTING: Eight of the largest primary care clinics in a managed care system. SAMPLE: 342 patients with cancer who reported pain that would not dissipate on its own or when treated by over-the-counter medication; approximately half had pain that was not cancer related. METHODS: Telephone interviews. MAIN RESEARCH VARIABLES: Patients' demographic characteristics, self-reported history and beliefs about pain and pain treatment, willingness to report pain and take pain medication, recent pain intensity, and administrative data on opioid prescriptions. FINDINGS: Patients' beliefs were not associated strongly with reporting pain or taking medication. Regression analyses revealed that patients' beliefs played a limited role in predicting recent pain intensity, whereas the providers' pain management practices seemed to have a far greater predictive role. Additionally, among patients with recent moderate to severe pain, the relationship between patients' beliefs and their history of pain and pain treatment further suggests that beliefs are likely to be formed, in part, as a consequence of the care they receive. Results did not depend on whether the cause of pain was related to cancer. CONCLUSION: Patients' beliefs were important barriers to effective pain management, either as direct or indirect determinants of pain. Providers' pain management practices were more likely to determine the level of pain relief achieved and the beliefs their patients came to hold based on their personal experiences. IMPLICATIONS FOR NURSING: Nursing interventions should examine the impact of evaluating patients' beliefs in conjunction with pain assessment on pain-related behaviors and pain relief, as well as the ability of patient educational efforts to strengthen accurate beliefs and enable patients to assert themselves when interacting with less knowledgeable providers. PMID- 15759074 TI - Impact of an intensive nursing education course on nurses' knowledge, confidence, attitudes, and perceived skills in the care of patients with cancer. AB - PURPOSE/OBJECTIVES: To evaluate the impact of a cancer nursing education course on RNs. DESIGN: Quasi-experimental, longitudinal, pretest/post-test design, with a follow-up assessment six weeks after the completion of the nursing education course. SETTING: Urban, nongovernment, cancer control agency in Australia. SAMPLE: 53 RNs, of whom 93% were female, with a mean age of 44.6 years and a mean of 16.8 years of experience in nursing; 86% of the nurses resided and worked in regional areas outside of the state capital. METHODS: Scales included the Intervention With Psychosocial Needs: Perceived Importance and Skill Level Scale, Palliative Care Quiz for Nurses, Breast Cancer Knowledge, Preparedness for Cancer Nursing, and Satisfaction With Learning. Data were analyzed using multiple analysis of variance and paired t tests. MAIN RESEARCH VARIABLES: Cancer nursing related knowledge, preparedness for cancer nursing, and attitudes toward and perceived skills in the psychosocial care of patients with cancer and their families. FINDINGS: Compared to nurses in the control group, nurses who attended the nursing education course improved in their cancer nursing-related knowledge, preparedness for cancer nursing, and attitudes toward and perceived skills in the psychosocial care of patients with cancer and their families. Improvements were evident at course completion and were maintained at the six-week follow-up assessment. CONCLUSIONS: The nursing education course was effective in improving nurses' scores on all outcome variables. IMPLICATIONS FOR NURSING: Continuing nursing education courses that use intensive mode timetabling, small group learning, and a mix of teaching methods, including didactic and interactive approaches and clinical placements, are effective and have the potential to improve nursing practice in oncology. PMID- 15759078 TI - Speed-accuracy tradeoff operator characteristics of endogenous and exogenous covert orienting of attention. AB - There is debate over the mechanisms that govern the orienting of attention. Some argue that the enhanced performance observed at a cued location is the result of increased perceptual sensitivity or preferential access to decision-making processes. It has also been suggested that these effects may be the result of trades in speed for accuracy on the part of the observers. In the present study, observers performed either an exogenous or an endogenous orienting of attention task under both normal instructions (respond as quickly and as accurately as possible) and speeded instructions that used a deadline procedure to limit the amount of time observers had to complete a choice reaction time (CRT) task. An examination of the speed-accuracy operating characteristics (SAOCs) yielded evidence against the notion that CRT precuing effects are due primarily to a tradeoff of accuracy for speed. PMID- 15759079 TI - Routine PSA testing: an analysis of the controversy concerning its use. AB - Prostate cancer is the leading cause of cancer in American males today. PSA screening has been used for over 10 years as an important diagnostic tool for the disease. Because of its lack of sensitivity and specificity, however, PSA testing should be used with caution. PMID- 15759081 TI - Clinical holistic medicine: chronic infections and autoimmune diseases. AB - The consciousness-based (holistic) medical toolbox might be useful in general practice and in cases of recurrent infections and chronic infection or inflammation. From our clinical experiences, there is hope for improvement from a number of diseases caused by disorders affecting the regulation of the immune system when the physician includes the holistic medical approach. Our scientific understanding of the connection between consciousness and cellular order is still limited. Consciousness-based holistic medicine removes (as explained by the holistic process theory of healing) the "blockages" in the tissues of the body and facilitates function and informational exchange of the cells of the body. Many blockages and repressed feelings in an area would imply "noise and disturbances" on the level of intercellular communications, which in turn means major difficulties for the cells of the immune system. For this they are totally dependent on the body information system, which the holistic treatment aims to recover. Processing the blockages increases the coherence of the cells and organism, thus increasing the intercellular flow of information in the area and thus strengthening the immune defense and healing the disease. The area of clinical holistic medicine is going through a rapid development and the toolbox of consciousness-based medicine is available for dealing with many diseases arising from disturbances in the regulation of the immune system. Holistic medicine has yet to be better explained scientifically and our proposed holistic cures have yet to be documented clinically. We invite the medical community to cooperate on this important challenge. PMID- 15759080 TI - Natural cytotoxicity receptors: pattern recognition and involvement of carbohydrates. AB - Natural cytotoxicity receptors (NCRs), expressed by natural killer (NK) cells, trigger NK lysis of tumor and virus-infected cells on interaction with cell surface ligands of these target cells. We have determined that viral hemagglutinins expressed on the surface of virus-infected cells are involved in the recognition by the NCRs, NKp44 and NKp46. Recognition of tumor cells by the NCRs NKp30 and NKp46 involves heparan sulfate epitopes expressed on the tumor cell membrane. Our studies provide new evidence for the identity of the ligands for NCRs and indicate that a broader definition should be applied to pathological patterns recognized by innate immune receptors. Since nonmicrobial endogenous carbohydrate structures contribute significantly to this recognition, there is an imperative need to develop appropriate tools for the facile sequencing of carbohydrate moieties. PMID- 15759082 TI - Clinical holistic medicine: chronic pain in the locomotor system. AB - Most pains from the locomotor system arise due to involuntary, chronic tensions in the muscles or other tissues. When the patient is motivated, the pain is easily cured in most of the cases by using the tools of consciousness-based medicine, primarily therapeutic touch, conversation, and coaching the patient in a positive philosophy of life. The pains are often caused by "blockages" that may cause problems other than just pain. Often it turns out that the blocked areas develop actual physical damage over time: a slipped disk in the back, articular degeneration, or osteoarthritis when the cartilage is affected, can often be explained in this way. Apparently, the exact areas where the blockage is situated cause cellular problems, disrupting cellular order. The holistic process theory of healing and the related quality of life theories state that return to the natural state of being is possible, whenever the person gets the resources needed for existential healing. The resources needed are "holding" in the dimensions of awareness, respect, care, acknowledgment, and acceptance with support and processing in the dimensions of feeling, understanding, and letting go of negative attitudes and beliefs. The preconditions for holistic healing are trust and the intention for the healing to take place. Case stories of holistic treatment of patients with chronic back pain, low back pain, muscle problems, knee pain, and symptoms of rheumatoid arthritis are discussed with exercises relevant for patients with these conditions in the holistic clinic. PMID- 15759083 TI - [Comparison of the minimal erythema dose for narrowband UV-B and broad spectrum UV-B using a new UV hand-held device]. AB - BACKGROUND: Phototherapy requires an individually adapted therapeutic regime that is based upon the minimal erythema dose for each patient. Whereas this is well established for UV-B broadband, so far no standardized equipment existed that allowed for determination of the MED for UV-B 311 nm narrowband irradiation. Thus, the starting dose for UV-B 311 nm is usually adjusted to the clinical skin type or the MED for broad-spectrum UV-B irradiation. PATIENTS AND METHODS: We determined the MED for UV-B 311 nm in a cohort of 110 healthy test persons with a newly constructed hand-held UV-light source and compared it to UV-B broadband MED. RESULTS: For UV-B broadband, the MEDs ranged from 46.2 to 210.9 mJ/cm(2) with an average value of 108.45 mJ/cm(2). For the MED of narrowband UV-B 311 nm, the values ranged from 300.9 to 1.386.0 mJ/cm(2) with an average value of 838.88 mJ/cm(2). Concerning the factor of the MED UV-B broad-spectrum related to MED UV B 311 nm narrowband, values from 4.01 to 13.68 with a average value of 7.91 resulted. No significant differences could be detected for the MED of UV-B broad spectrum and narrowband in correlation to age, sex or clinical skin type. CONCLUSION: Our data emphasize the presence of extensive inter-individual differences in the MED of both UV-B-spectra and point to the necessity to test patients before the first irradiation. PMID- 15759084 TI - Atlantic cod trypsins: from basic research to practical applications. AB - Atlantic cod trypsin I is an appropriate representative of the traditionally classified cold-adapted group I trypsins, and the recombinant form of cod trypsin Y is the only biochemically characterized member of the novel group III trypsins. Trypsin Y is adapted to lower temperatures than all other presently known trypsins. This review describes the basic characteristics of and practical uses for trypsins of Atlantic cod, as well as those of other organisms. Overexpression of the recombinant forms of cod trypsins I and Y in microorganisms is explained as well as the advantages of using site-directed mutagenesis to increase their stability toward autolysis and thermal inactivation. Trypsins appear to play a key role in the nutrition and development of marine fish. We discuss the potential use of cod trypsins as biomarkers to evaluate the nutritional status of cod larvae and describe the industrial applications of cod trypsin I and other trypsins. PMID- 15759085 TI - Molecular phylogenetics of gadidae and related gadiformes based on mitochondrial DNA sequences. AB - Mitochondrial DNA sequences of selected regions of the small subunit and large subunit ribosomal RNAs and cytochrome b genes were analyzed for 10 gadid species, representing 8 genera within Gadidae, and 10 species representing 5 other gadiform families. Phylogenetic analyses revealed that Gadiculus is the most basal gadid genus, and that Trisopterus and Micromesistius constitute a relatively basal clade. Lotidae was identified as the family most closely related to Gadidae. Estimation of divergence times indicated that the most ancient Gadidae split between Gadiculus and the remaining gadid genera occurred about 20 million years ago. The clade including the most recent species (Gadus, Boreogadus, Merlangius, Melanogrammus, and Pollachius) diverged from the Trisopterus/Micromesistius clade approximately 12 million years ago. PMID- 15759086 TI - Proteolytic extraction of salmon oil and PUFA concentration by lipases. AB - Commercial proteases (Alcalase, Neutrase, and Flavourzyme) were tested for their ability to release the oil content of marine by-products (salmon heads). The amount of oil (17%) obtained after 2 hours was close to that obtained by the chemical extraction method (20%). Lipolysis of the oil was carried out with Novozym SP398 to obtain a mixture of free fatty acids and acylglycerols (24 hours 45% hydrolysis). The mixture was filtered on a hydrophobic membrane to discriminate between high melting saturated fatty acids and low melting acylglycerols. The sum of total polyunsaturated fatty acids increased from 41.6% in the crude oil to 46.5% in the permeate. The docosahexaenoic acid content increased from 9.9% to 11.6%, and the eicosapentaenoic acid changed from 3.6% to 5.6%. Data from differential scanning calorimetry DSC and from thin layer chromatography coupled with flame ionization detection (TLC-FID) differed significantly between permeate and retentate. A re-esterification of the free fatty acids in the permeate with Lipozyme IM was carried out to increase the amount of long chain acylglycerols. PMID- 15759087 TI - Cellular, histomorphologic, and clinical characteristics of a new octyl-2 cyanoacrylate skin adhesive. AB - Short-chained cyanoacrylates have been used for many years for topical skin closure. Toxic effects in cell culture of a new long-chained octyl-2 cyanoacrylate tissue adhesive are compared with those of short-chained ethyl-2- and butyl-2-cyanoacrylates. Two cellular tests were used: the agar overlay test and the MTT test. An in vitro test using copper plates coated with the three types of cyanoacrylates serves for evaluating the stability of polymerized skin adhesives. Bilateral neck skin incisions in Goettingen miniature pigs were glued on one side with Dermabond. On the other side, conventional sutures were applied. After the pigs were killed, the resulting skin samples were tested for the tensile strength of their wound stability. Samples of pig dermis were exemplarily and histomorphologically characterized. A clinical examination after submandibular lymph node dissection should examine the application in humans. Cell culture tests were used to show the toxic effects of the three cyanoacrylates. In a copper test, octyl-2-cyanoacrylate was more stabile than ethyl- and butyl-cyanoacrylates. Breaking strength was 30% lower 28 days after operation with the new product than with sutures. In electron microscopy, octyl-2 cyanoacrylate showed no disadvantages with regard to tissue regeneration and no histotoxicity. For plastic surgery, this new topical skin adhesive is a real alternative with attractive results, as compared with conventional suture. PMID- 15759088 TI - Late recurrent capsular hematoma after augmentation mammaplasty: case report. AB - Late capsular hematoma after the implantation of silicone breast prostheses for augmentation mammaplasty is an extremely rare complication. The case of a patient in whom two late intracapsular hematomas occurred two times after augmentation mammaplasty with textured silicone implants is reported. Magnetic resonance imaging clearly showed fluid accumulated within the capsule. No identifiable etiology could be demonstrated, although the breakdown of an eroded capsular artery is suspected, caused by friction between the rough surface of the textured prosthesis and the fibrous capsule. Two hematomas in a single patient have not been described previously in the literature. PMID- 15759089 TI - Evaluation of cutaneous abdominal wall sensibility after abdominoplasty. AB - Changes in cutaneous sensibility are common after diverse plastic surgical procedures. Although prior studies examined aesthetic results, combined procedures, and new abdominoplasty techniques, few examined the effect of undermining on cutaneous sensibility. This study aimed to analyze and quantify cutaneous sensibility after classic abdominoplasty. Two groups of patients were studied: a control group of 10 patients without surgery and another group of 25 patients who had undergone classic abdominoplasty. The abdominal wall surface was divided into nine regions. Pain sensibility was evaluated by tests with needles, and thermal sensibility by test tubes containing hot and cold water. Superficial tactile sensibility was tested using the Pressure-Specified Sensory Device (PSSD), which is capable of determining the cutaneous pressure threshold. Statistical analysis was conducted using Student's 't-test'. The results showed a decrease in the three types of sensibility. Statistical analysis showed a significant difference (p < 0.05) between the groups for all regions tested. The centermost regions of the abdominal wall presented the highest index of analgesia and thermal anesthesia, as well as higher cutaneous pressure thresholds. PMID- 15759091 TI - A new simple method used to prepare fat for injection. AB - Various clinical and experimental studies on harvesting techniques, donor sites, preparation, and preservation of fat for injection imply that there is no universal agreement on an ideal methodology. Unpredictable and irregular resorption of fat leads to uncertainty about volume maintenance of the transplanted fat necessitating repeat procedures. In this report, the authors present their 5-year clinical experience with facial fat grafting by isolated and combined procedures using a simple method that they developed for fat preparation. This technique includes fat harvesting with syringe aspiration and filtration of the fat aspirate instead of centrifugation. The idea for filtration was inspired by a yogurt concentration method used in Anatolia. The authors also aimed to increase fat survival by using multiple fat injections of minimal amounts into different levels. PMID- 15759092 TI - Poland syndrome: description of an atypical variant. AB - Poland syndrome comprises a unilateral absence of the large pectoral muscle, ipsilateral symbrachydactyly, and occasionally other malformations of the anterior chest wall and breast. The condition is more frequent among males, and usually occurs on the right hemithorax in the unilateral form. The syndrome is believed to be caused by a genetic disorder that reduces the embryonal circulation in the interior chest artery: the stronger the interaction, the more severe the pathology. This article analyzes an unusual pathologic case in which the 17-year-old patient lacked the large pectoral muscle on the left side, but showed no arterial alteration. This case raises questions as to the true pathogenesis of this syndrome. PMID- 15759093 TI - Power-assisted lipoplasty versus traditional suction-assisted lipoplasty: comparative evaluation and analysis of output. AB - Power-Assisted lipoplasty (PAL) is a new method introduced to simplify and standardize surgical results of traditional suction-assisted lipoplasty (SAL). Comparative studies already have demonstrated PAL to be a handy, atraumatic, time and fatigue-sparing technique. The authors performed a pilot study to compare the output capacity of PAL and SAL in 15 healthy female patients. The general and the specific per area outputs over 1 min from symmetrical areas were assessed. The results confirmed the efficiency of PAL (17.41% increase in average specific area output, as compared to SAL) and the distribution of fat in major "fat storage" areas such as the abdomen and the anterior thigh. Specific per area output indicated a higher suction capacity for PAL in all areas except the inner thigh. The efficiency of PAL seemed to be less influenced than that SAL by fat distribution. PMID- 15759094 TI - New observations in liposuction-only breast reduction. AB - BACKGROUND: Breast reduction surgery is known to be more effective than medications, diet, or support bras in relieving the suffering from excessively large breasts. Liposuction-only breast reduction, without surgery, is effective for eliminating symptoms and significantly reduces scarring and complications. The procedure allows women to maintain nipple sensation and the ability to breast feed. The author describes his technique, results, and new observations over a 2 year period. METHODS: The results from the charts and photos of all patients in the study group were retrospectively reviewed. RESULTS: The patients were very satisfied with the results of their treatment. New observations are detailed. Complications were minimal. No cases of increased ptosis resulted from suction. CONCLUSIONS: Breast liposuction is an excellent technique that reduces scars, complications, recovery, and costs. It is a technique that should be readily available to women in every society. PMID- 15759095 TI - Mycobacterium cheloneae infection after breast augmentation. AB - Augmentation mammaplasty is one of the most commonly performed plastic surgery procedures. Infection, still one of the most feared complications of the procedure, usually is caused by skin commensal organisms. A wide variety of other organisms also may be responsible for these postoperative infections, including atypical mycobacteria. A case of a prosthetic breast implant infected with Mycobacterium cheloneae is presented, and the presenting signs and symptoms, diagnosis, and treatment options of such an infection are discussed. The importance of promptly isolating the pathogen for potential salvage of the prosthesis also is stressed, as well as the operative intervention necessary when conservative therapy fails. PMID- 15759096 TI - Adverse reactions to injectable soft tissue permanent fillers. AB - BACKGROUND: Synthetic injectable facial fillers with a permanent effect are widely atoxic and nonimmunogenic, but they differ with respect to composition and in chemical and biologic characteristics. Yet, they all act as foreign bodies in the tissues eliciting a host response that try to remove the gel. Inflammatory nodules may develop at the sites of injection-for some fillers, many years later, for others, not. Why is that? METHODS: Biopsies were contributed by various plastic surgeons from Europe and Australia after requests were made at international congresses and workshops. The study was based on (a) 5 biopsies from unreactive tissue obtained at different times after injection of polyacrylamide hydrogel (Aquamid); (b) 28 biopsies from intermediate or late inflammatory nodules after injection of polyacrylamide hydrogel (Aquamid) (20 cases), a hyaluronic acid-polyhydroxyethylmethacrylate/ethylmethacrylate gel (Dermalive) (2 cases), and a gel consisting of polylactic acid in mannitol/carbomethoxycellulose (New-Fill) (6 cases); and (c) a review of the literature on adverse reactions after injection with permanent fillers. RESULTS: Clinically unreactive tissues after injection with Aquamid showed modest or no host reaction. Inflammatory nodules showed an increased foreign body reaction and a bacterial infection after injection with Aquamid, and a combination of moderate foreign body reaction, fibrosis, and in some cases also bacterial infection after injection with Dermalive and New-Fill. According to the literature, inflammatory nodules occur no later than 1 year after injection with polyacrylamide hydrogel, but up to 6 years after injection of combination gels (Artecol), and up to 28 years after injection of silicone gel. CONCLUSIONS: Inflammatory nodules are likely to be caused by a low-grade infection maintained within a biogfilm surrounding the hydrophobic silicone gel and the combination gels. Aquamid gel may prevent formation of a biofilm through its high water-binding capacity, explaining why late inflammatory nodules are not seen after injection of this polyacrylamide hydrogel product. PMID- 15759098 TI - [Teledermatology versus consultations--a comparative study of 120 consultations]. AB - Dermatology fulfills the prerequisites for telemedicine. An important application of telemedicine might be the field of dermatologic consultations. In this comparative study images of skin disease were taken of 120 patients hospitalized in the University Hospitals of Tuebingen to answer the following questions: (1) are the preconditions in daily routine given for teledermatology, (2) is there adequate agreement between the diagnoses reached in dermatologic consultations and with teledermatology, and (3) can the images be utilized for teaching purposes. Patient acceptance was very good and the images captured with a digital camera were easily obtained. The results of intraobserver analysis for the two teledermatologists without any knowledge of the patients' history were 70.2% and 46.4%, respectively, and with knowledge of the history 76.6% and 64.3%. The results of interobserver analysis without any knowledge of the patients' history were 46.4% and 57.2% and with knowledge of the history 64.3% and 66%, respectively. With the reduction of the image quality, reduced reliability of the diagnoses was observed. Seven of ten images could be used for teaching purposes. It was demonstrated that in dermatology telemedicine is applicable in many, but not in all patients who are referred for dermatologic consultations. PMID- 15759099 TI - Novel biochemistry: post-translational protein splicing and other lessons from the school of antigen processing. AB - In the past 15 years, the molecular identification of antigens that can mediate the killing of tumor cells by T cells has been vigorously pursued. Molecular identification of tumor-associated antigens not only provided the means to activate or monitor anti-tumor immunity, but also gave insights into new and unexpected biochemical processes that are taking place within cells. Post translational splicing, a phenomenon previously identified only in lower organisms or plants, has recently been added to the list of atypical processes generating proteins in humans. The proteasome, whose main function is to degrade intracellular proteins, appears to catalyze this splicing reaction. The discovery of post-translational splicing has immediate and important implications for the complexity of the major histocompatibility complex (MHC) class I peptide repertoire and for the immune recognition of self- and foreign peptides. PMID- 15759101 TI - Tyrosinaemia type I--de novo mutation in liver tissue suppressing an inborn splicing defect. AB - Many patients with tyrosinaemia type 1 have a mosaic pattern of fumarylacetoacetase (FAH) immunopositive or immunonegative nodules in liver tissue. This phenomenon has been explained by a spontaneous reversion of the mutation in one allele to a normal genotype, but only a few nodules have been examined. We now report on a Norwegian patient, compound heterozygous for the mutations IVS12g(+5)-->a and G(1009-->)A, with liver mosaicism, but with an immunopositive nodule in which both primary mutations were intact. In the immunopositive hepatocytes of this nodule, genetic analyses showed a new mutation, C(1061-->)A, 6 bp upstream of the primary mutation IVS12g(+5)-->a in the FAH gene. The splicing defect caused by the primary mutation is most likely suppressed by the new mutation due to improvement of the splicing site. In the same liver we demonstrate another nodule of regenerating immunopositive tissue due to reversion of one of the primary mutations to a normal genotype. Together with the original cells this makes a triple mosaicism of hepatocytes with one, two or three point mutations in the FAH gene. PMID- 15759100 TI - Therapeutic gene silencing in neurological disorders, using interfering RNA. AB - The development of interfering RNA (RNAi) from a naturally occurring phenomenon to a tool for mediating highly specific gene silencing provides an exciting prospect as a novel therapeutic strategy for a wide range of disorders. Although the efficacy of RNAi as a research tool for analysing gene function has been well demonstrated in several cell types, the therapeutic potential of RNAi-mediated gene silencing has only recently started to be investigated. Several neurodegenerative disorders provide particularly suitable candidates for RNAi based therapy; however, many hurdles preclude the success of therapeutic application. These include the challenge of delivering active RNAi molecules to the specific target cell populations where they are required and appropriate regulation of gene suppression, such as to maintain a long-lasting therapeutic effect. Furthermore, for safety reasons, off-target effects should be minimised. Here we review the advancement of RNAi technology for therapeutic application and highlight the potential of targeted gene silencing for the treatment of neurodegenerative diseases. PMID- 15759102 TI - Oxidative stress, ER stress, and the JNK pathway in type 2 diabetes. AB - Pancreatic beta-cell dysfunction and insulin resistance are observed in type 2 diabetes. Under diabetic conditions, oxidative stress and ER stress are induced in various tissues, leading to activation of the JNK pathway. This JNK activation suppresses insulin biosynthesis and interferes with insulin action. Indeed, suppression of the JNK pathway in diabetic mice improves insulin resistance and ameliorates glucose tolerance. Thus, the JNK pathway plays a central role in pathogenesis of type 2 diabetes and may be a potential target for diabetes therapy. PMID- 15759103 TI - A novel therapeutic strategy for polyglutamine diseases by stabilizing aggregation-prone proteins with small molecules. AB - Polyglutamine diseases, such as Huntington disease (HD) and spinocerebellar ataxia 1 and 3, are autosomal dominant neurodegenerative disorders. They are caused by CAG trinucleotide repeat expansions that are translated into abnormally long polyglutamine tracts. One of the pathological hallmarks in polyglutamine diseases is the formation of intranuclear inclusions of polyglutamine-containing proteins in the brain. Although causal relationships between polyglutamine aggregation and cellular toxicity are much debated, inhibition of the polyglutamine-mediated protein aggregation may provide treatment options for polyglutamine diseases. However, the extreme insolubility of expanded polyglutamines makes it difficult to prepare polyglutamine-containing proteins on a large scale and to search for aggregation inhibitors by in vitro high throughput screening. To overcome this we developed a novel in vitro model system for polyglutamine diseases using myoglobin as a host protein. We searched for small molecules that inhibit polyglutamine-mediated aggregation by in vitro screening with a mutant myoglobin containing a 35 polyglutamine repeat. The screening assay revealed that disaccharides have a potential to inhibit polyglutamine-induced protein aggregation and to increase survival in a cellular model of HD. Oral administration of trehalose, the most effective disaccharide in vitro, decreased polyglutamine aggregates in the cerebrum and liver, improved motor dysfunction and extended life span in a transgenic mouse model of HD. In vitro experiments suggest that the beneficial effects of trehalose result from its ability to bind and stabilize polyglutamine-containing proteins. The lack of toxicity and high solubility, coupled with its efficacy upon oral administration, make trehalose promising as a therapeutic drug or lead compound for the treatment of polyglutamine diseases. The stabilization of aggregation-prone proteins with small molecules is an attractive strategy because it can block the initial stage of the disease cascade. In addition, this therapeutic approach could be applied not only to polyglutamine diseases but also to a wide variety of misfolding induced diseases. PMID- 15759104 TI - Mast cell deficient and neurokinin-1 receptor knockout mice are protected from stress-induced hair growth inhibition. AB - Despite the lack of insight on distinct mediators in the skin orchestrating the pathophysiological response to stress, hair loss has often been reported to be caused by stress. Recently we revealed the existence of a "brain-hair follicle axis" by characterizing the neurokinin (NK) substance P (SP) as a central element in the stress-induced threat to the hair follicle, resulting in premature onset of catagen accompanied by mast cell activation in the skin. However, our understanding of possible SP-mast cell interactions in the skin in response to stress was limited since the receptor by which SP activates skin mast cells and the extent of mast cell mediated aggravation of SP remained to be elucidated. We now employed NK-1 receptor knockout mice (NK-1R(-/-)) and mast cell deficient W/W(v) mice and observed that stress-triggered premature induction of catagen and hair follicle apoptosis does not occur in NK1(-/-) and W/W(v) mice. Furthermore, the activation status of mast cells was less in stressed NK1(-/-) mice than in wild-type control. Additionally, stress-induced upregulation of SP positive nerve fibers was absent in both NK-1R and W/W(v) mice. These results indicate that the cross-talk between SP and mast cell activation via NK-1R appears to be the most important pathway in the regulation of hair follicle cycling upon stress response. PMID- 15759105 TI - Genetics of resistance to septoria tritici blotch in the Portuguese wheat breeding line TE 9111. AB - We report the genetics of resistance of the Portuguese wheat breeding line TE 9111 to septoria tritici blotch (STB), which is caused by Mycosphaerella graminicola. TE 9111 is the most resistant line known in Europe and combines isolate-non-specific, partial resistance with several isolate-specific resistances. We show that, in addition to high levels of partial resistance to STB, TE 9111 has a new gene for resistance to M. graminicola isolate IPO90012, named Stb11, that maps on chromosome 1BS, the Stb6 gene for resistance to isolate IPO323 and, probably, the Stb7 gene for resistance to isolate IPO87019. All of these genes are closely linked to microsatellite markers, which can be used for marker-assisted selection. TE 9111 may therefore be a valuable source of resistance to STB for wheat breeding, especially in Mediterranean environments. PMID- 15759106 TI - Glucagon-like peptide-1 mediates the therapeutic actions of DPP-IV inhibitors. PMID- 15759107 TI - Mean HbA1c over 18 years predicts carotid intima media thickness in women with type 1 diabetes. AB - AIMS/HYPOTHESIS: Intima media thickness (IMT) of the common carotid artery (CCA) is a validated surrogate marker of early atherosclerosis. The aim of our study was to assess the association between IMT in CCA and long-term mean HbA1c in type 1 diabetes. We also elucidated the association between carotid IMT and preclinical coronary atherosclerosis. METHODS: In 39 individuals with type 1 diabetes, HbA1c was measured prospectively over 18 years. The IMT examinations were performed with high-resolution ultrasound. The association between carotid IMT and preclinical coronary atherosclerosis (assessed by intravascular ultrasound [IVUS]) was tested in 29 of the patients. RESULTS: Mean HbA1c over 18 years was 8.2% (range: 6.6-11.3%). Mean age at follow-up after 18 years was 43 years and mean duration of diabetes was 30 years. IMT was significantly higher in diabetic patients than in an age- and sex-matched reference population. The IMT values were at the same level as for controls who were 20 years older. In women, HbA1c was significantly associated with mean average CCA IMT (r2=0.77, p<0.0001 when adjusted for age), whereas there was no significant association for men. Among women, a significant association was also found between carotid IMT and the percentage of coronary vessel area stenosis (r=0.65, p=0.03). CONCLUSIONS/INTERPRETATION: The present findings suggest an important role of long-term hyperglycaemia in the development of atherosclerosis, especially in women with type 1 diabetes. Type 1 diabetes patients have earlier development of, and more advanced, atherosclerosis compared with an age- and sex-matched reference population. In women, carotid IMT reflects preclinical coronary atherosclerosis. PMID- 15759108 TI - Prospective incidence study of diabetes mellitus in New Zealand children aged 0 to 14 years. AB - AIMS/HYPOTHESIS: The aim of this study was to establish the incidence of type 1 and type 2 diabetes mellitus in children aged 0-14 years. METHODS: The New Zealand Paediatric Surveillance Unit sought monthly reporting of diabetes mellitus cases from paediatricians. All resident children aged below 15 years (1996 census risk population 832,000) who met the criteria for diagnosis of diabetes mellitus from 1 January 1999 to 31 December 2000 were included. The average annual incidence of type 1 and type 2 diabetes was calculated, as were incidence rates according to age, sex, region, ethnicity and season. Case ascertainment was estimated using hospital admission data. RESULTS: There were 315 valid reports of new cases of diabetes. Of these, 298 (94.6%) had type 1 diabetes, 12 (3.8%) had type 2 diabetes and five had other specified types of diabetes. The average annual incidence of type 1 diabetes was 17.9/100,000 (95% CI: 15.9-20/100,000). Children in the South Island had a 1.5-fold higher incidence than children in the North Island, which was largely accounted for by the variation in incidence with ethnicity, in that the European rate was 4.5 times higher than the Maori rate. The average annual incidence of type 2 diabetes was 0.84/100,000 (95% CI: 0.37-1.26/100,000). Estimated case ascertainment rate was 95.2%. CONCLUSIONS/INTERPRETATION: Type 1 diabetes incidence has doubled over the past three decades. The geographical differences previously described have persisted, and are largely explained by the ethnic variation in incidence. This population includes young adolescents with type 2 diabetes. These findings are in keeping with international trends. PMID- 15759110 TI - Metabolic syndrome and coronary heart disease in South Asians, African-Caribbeans and white Europeans: a UK population-based cross-sectional study. AB - AIMS/HYPOTHESIS: The aim of this study was to study differences in the prevalence of the metabolic syndrome and its associations with prevalent CHD according to ethnicity and sex. METHODS: We performed a combined analysis of two population based cross-sectional studies conducted between 1988 and 1991 that followed identical protocols. Participants (aged 40-69 years) comprised 2,346 Europeans (76% male), 1,711 South Asians (83% male) and 803 African-Caribbeans (57% male) resident in west London. Fasting blood, overnight urine collection, clinical and anthropometric measurements were performed. Clinical history or major ECG changes defined prevalent CHD. The metabolic syndrome was defined according to the criteria recommended by the World Health Organization (WHO) and the National Cholesterol Education Programme (NCEP). RESULTS: The prevalence of the metabolic syndrome was highest in South Asians (WHO, men 46%, women 31%; NCEP, men 29%, women 32%) and lowest in European women (WHO, 9%; NCEP, 14%). The prevalence of CHD was 10% in South Asian men, 9% in European men, 5-6% in African-Caribbeans and European women, and 2% in South Asian women. The metabolic syndrome was associated with prevalent CHD in European men [NCEP, odds ratio (OR)=1.6, 95% CI 1.2-2.4; WHO, OR=1.7, 95% CI 1.2-2.5] and South Asian men (NCEP, OR=2.1, 95% CI 1.5-3.1; WHO, OR=1.6, 95% CI 1.1-2.3). Associations with CHD were weaker in African-Caribbeans and were inconsistent among European women. CONCLUSIONS/INTERPRETATION: The current definitions of the metabolic syndrome give an inconsistent picture of cardiovascular disease risk when applied to different ethnic groups within the UK. Prospective studies are needed to validate workable ethnic-specific definitions. PMID- 15759109 TI - Efficient restricted gene expression in beta cells by lentivirus-mediated gene transfer into pancreatic stem/progenitor cells. AB - AIMS/HYPOTHESIS: Gene transfer into pancreatic beta cells, which produce and secrete insulin, is a promising strategy to protect such cells against autoimmune destruction and also to generate beta cells in mass, thereby providing a novel therapeutic approach to treat diabetic patients. Until recently, exogenous DNA has been directly transferred into mature beta cells with various levels of success. We investigated whether exogenous DNA could be stably transferred into pancreatic stem/progenitor cells, which would subsequently differentiate into mature beta cells expressing the transgene. METHODS: We designed transplantation and tissue culture procedures to obtain ex vivo models of pancreatic development. We next constructed recombinant lentiviruses expressing enhanced green fluorescent protein (eGFP) under the control of either the rat insulin promoter or a ubiquitous promoter, and performed viral infection of rat embryonic pancreatic tissue. RESULTS: Embryonic pancreas infected with recombinant lentiviruses resulted in endocrine cell differentiation and restricted cell type expression of the transgene according to the specificity of the promoter used in the viral construct. We next demonstrated that the efficiency of infection could be further improved upon infection of embryonic pancreatic epithelia, followed by their in vitro culture, using conditions that favour endocrine cell differentiation. Under these conditions, endocrine stem/progenitor cells expressing neurogenin 3 are efficiently transduced by recombinant lentiviral vectors. Moreover, when eGFP was placed under the control of the insulin promoter, 70.4% of the developed beta cells were eGFP-expressing cells. All of the eGFP-positive cells were insulin-producing cells. CONCLUSIONS/INTERPRETATION: We have demonstrated that mature rat pancreatic beta cells can be stably modified by infecting pancreatic stem/progenitor cells that undergo endocrine differentiation. PMID- 15759111 TI - Reduced whole-body lipid oxidation is associated with insulin resistance, but not with intramyocellular lipid content in offspring of type 2 diabetic patients. AB - AIMS/HYPOTHESIS: Intramyocellular lipid accumulation and insulin resistance are thought to be due to reduced lipid oxidation in a human model of high risk of developing type 2 diabetes. METHODS: We studied 32 offspring of type 2 diabetic parents and 32 control individuals by means of DXA, indirect calorimetry, insulin clamp and 1H MRS of the calf muscles, and differences between and within study groups were analysed before and after segregation by quartiles of fasting lipid oxidation. RESULTS: In comparison with control subjects, the offspring showed impaired insulin sensitivity, which was associated with higher fasting intramyocellular lipid content (Spearman's rho -0.35; p=0.04), but fasting lipid oxidation did not differ between groups (1.21+/-0.46 vs. 1.25+/-0.37 mg.kg(-1) lean body mass per min; p=0.70). Nevertheless, offspring in the lowest quartile of lipid oxidation had the most severe impairment of insulin sensitivity and a strong association was shown between lipid oxidation and insulin sensitivity within quartiles (Spearman's rho 0.47; p=0.01); this was not observed within the control group (Spearman's rho 0.13; p=0.47). Intramyocellular lipid content was not significantly different within quartiles of lipid oxidation in either of the groups. CONCLUSIONS/INTERPRETATION: Insulin sensitivity improved across increasing quartiles of fasting lipid oxidation in the offspring group, but remained constant in the control group, supporting the hypothesis that impaired fat oxidation is a primary pathogenic factor of insulin resistance in people with a genetic background for type 2 diabetes. Despite their association with impaired insulin sensitivity, soleus and tibialis anterior intramyocellular lipid content remained constant across increasing quartiles of fasting lipid oxidation within both groups. PMID- 15759114 TI - A novel tumor necrosis factor (TNF) gene present in tandem with theTNF-alpha gene on the same chromosome in teleosts. AB - Tumor necrosis factors (TNFs) are pleiotropic cytokines implicated in inflammation, apoptosis, cell proliferation, and a general stimulation of immune system. Although the TNF/lymphotoxin (LT) locus is present in the MHC region of the chromosome in mammals, no such locus has been described from teleosts. Furthermore, in teleosts only the TNF-alpha-like gene has been cloned and analyzed. In this study, for the first time in teleosts, a similar TNF locus has also been found in zebrafish and fugu. This locus harbors a new TNF ligand in tandem with TNF-alpha gene. The fugu and zebrafish TNF locus harbors TNF-alpha and TNF-N genes that are 7.5 kb and 19 kb apart from each other, respectively. Several genes associated with the TNF/LT locus in mammals are found conserved in fish as well. Fish TNF-alpha genes cloned possess a transmembrane domain, two conserved cysteines, and a conserved TNF family signature with a four-exon and three-intron genomic structure. The newly identified TNF-N genes from teleosts are present upstream of TNF-alpha genes in the same transcriptional orientation. However, the coding regions of these genes span into five and four exons in fugu and zebrafish, respectively. The transcription sites in the promoter regions of teleost TNF ligands are fairly conserved in comparison to mammals. Expression analysis indicates a constitutive expression of both the genes in fugu, while differential expression of TNF ligands is seen in zebrafish tissues. PMID- 15759115 TI - Autologous tandem transplantation in patients with primary progressive or relapsed/refractory lymphoma. AB - Patients with primary progressive or refractory Hodgkin's disease (HD) or aggressive non-Hodgkin's lymphoma (NHL) have a particularly poor prognosis. Here we report the results of autologous tandem transplantation in these patients. Patients aged 18-55 years with primary progressive or refractory relapsed HD and aggressive NHL were included. Patients received high-dose etoposide (2000 mg/m(2)) followed by peripheral blood stem cell harvest (PBSC). The first high dose chemotherapy (TMC) consisted of thiotepa (750 mg/m(2)), mitoxantrone (40 mg/m(2)), and carboplatin (990 mg/m(2)). Patients with no change (NC), partial remission (PR), or complete remission (CR) after TMC then received BEAM with carmustine (300 mg/m(2)), etoposide (1200 mg/m(2)), cytarabine (1600 mg/m(2)), and melphalan (140 mg/m(2)). Patients with bulky disease (>5 cm) or residual lymphoma received involved field radiotherapy. Twenty-five patients were included (HD=10, NHL=15, median age 34 years). Two patients with HD achieved a CR and five patients a PR [response rate (RR) 70%]. Three patients (30%) experienced treatment failure including two deaths due to peritransplant complications. Five patients with aggressive NHL were in CR and two patients in PR (RR 46%). Of the eight patients (56%) with treatment failure, three had progressive disease and five died from peritransplant complications. Freedom from treatment failure (FFTF) and overall survival (OS) for all patients after 12 months was 28% and 40%, respectively. Tandem HDCT followed by autologous stem cell transplantation (ASCT) offers a chance of cure in these poor prognostic patients, but is associated with risks. PMID- 15759116 TI - Organization of assembly factors Cbp3p and Cbp4p and their effect on bc(1) complex assembly in Saccharomyces cerevisiae. AB - The bc(1) complex (complex III) of Saccharomyces cerevisae is composed of ten subunits that are assembled in the inner mitochondrial membrane. Cbp3p and Cbp4p are two mitochondrial proteins which are postulated to act as chaperones in bc(1) complex formation. Here, we show by blue native PAGE that cbp3Delta and cbp4Delta mutants are disturbed in complex III assembly and accumulate intermediate-sized forms of the complex. Moreover, deletion of CBP3 interferes with the formation of complex III/IV supracomplexes. Our studies show that Cbp3p and Cbp4p interact and are present in high-molecular-weight complexes, some of which might represent intermediates of complex III assembly. Overexpression of Cbp4p cannot substitute for the function of Cbp3p, but high-level expression of Cbp3p can partially compensate for the lack of Cbp4p. The finding that mitochondria of cbp3Delta and cbp4Delta mutants exhibit a wild-type lipid composition favors the idea that Cbp3p and Cbp4p are specific assembly factors for complex III rather than components of the mitochondrial lipid metabolism. PMID- 15759117 TI - Aluminum-induced cell death of barley-root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production. AB - The function of root border cells (RBC) during aluminum (Al) stress and the involvement of oxalate oxidase, peroxidase and H(2)O(2) generation in Al toxicity were studied in barley roots. Our results suggest that RBC effectively protect the barley root tip from Al relative to the situation in roots cultivated in hydroponics where RBC are not sustained in the area surrounding the root tip. The removal of RBC from Al-treated roots increased root growth inhibition, Al and Evans blue uptake, inhibition of RBC production, the level of dead RBC, peroxidase and oxalate oxidase activity and the production of H(2)O(2). Our results suggest that even though RBC actively produce active oxygen species during Al stress, their role in the protection of root tips against Al toxicity is to chelate Al in their dead cell body. PMID- 15759118 TI - Achieving desired plant growth regulator levels in liquid plant tissue culture media that include activated carbon. AB - This paper is part of a series considering the impact of activated carbon (AC) on the composition of plant tissue culture media. Using liquid culture media for initiation of Norway spruce embryogenic tissue and eight different ACs, we present a method for achieving target plant growth regulator (PGR) levels in AC containing medium based on sorption isotherms for individual PGRs. Linear relationships were found between PGR adsorption and specific BET (Brunauer, Emmett, Teller theory) surface area and specific total pore volume of AC. When using a new AC, this linear relationship allows one to achieve multiple PGR levels similar to historic levels through adjustment of the mass of AC based on its relative BET surface area or relative total pore volume. Target levels of PGRs and an initiation success similar to that in medium without AC were achieved with several different AC types when AC mass was adjusted on the basis of pore volume. PMID- 15759119 TI - In vitro mutation and selection of doubled-haploid Brassica napus lines with improved resistance to Sclerotinia sclerotiorum. AB - This paper describes a new protocol to develop doubled-haploid (DH) Brassica napus lines with improved resistance to Sclerotinia sclerotiorum. In this protocol, haploid seedlings derived from microspore cultures of B. napus were used to produce haploid calli for in vitro mutation-selection. For routine screening, mutation was induced by EMS (ethylmethane sulfonate) or occurred spontaneously, and screening for resistant mutants occurred on media with added oxalic acid (OA) as a selection agent. In tests with selected lines, the optimal concentration of EMS for mutation was determined to be 0.15%, and the optimal concentration of OA for in vitro screening was 3 mmol/l (half lethal dose was 3.1 mmol/l) for the first cycle of screening. There was an accumulated effect of OA toxicity on calli over two cycles of screening, but the growth and capacity of the surviving calli for regenerating seedlings were not affected by OA. Of the 54 DH lines produced from the in vitro mutation-selection, two DH lines of resistant mutants, named M083 and M004, were selected following seedling and glasshouse tests. The resistance of M083 and M004 to S. sclerotiorum following tests with both mycelial inoculum and OA was greater than that of their donor lines and the resistant control Zhongyou 821. In both glasshouse and field disease nurseries, disease indices on M083 and M004 were less than 50% of those of the control. The time required for M083 and M004 to mature was 14 days and 10 days shorter, respectively, than that of their donor lines. Furthermore, M083 had more pods per inflorescence, a greater 1,000 seed weight and higher yield than its donor line. Random amplified polymorphic DNA characterisation showed that M083 had DNA band patterns that differed from its donor line. PMID- 15759120 TI - Molecular cloning and expression analysis of the cell-wall invertase gene family in rice (Oryza sativa L.). AB - Cell-wall invertase (CIN) catalyzes the hydrolysis of sucrose into glucose and fructose for the supply of carbohydrates to sink organs via an apoplastic pathway. To study the CIN genes in rice (Oryza sativa L.), we isolated cDNA clones showing amino acid similarity to the plant cell wall invertase proteins from a search of rice sequence databases. Profile analyses revealed that the cloned genes are expressed in unique patterns in various organs. For example, transcripts of OsCIN1, OsCIN2, OsCIN4, and OsCIN7 were detected in immature seeds whereas OsCIN3 gene expression was flower-specific. Further transcript analysis of these genes expressed in developing seeds indicated that OsCIN1, OsCIN2, and OsCIN7 might play an important role involving sucrose partitioning to the embryo and endosperm. Sucrose, a substrate of CINs, induced the accumulation of OsCIN1 transcripts in excised leaves and OsCIN2 in immature seeds, while the level of OsCIN5 was significantly down-regulated in excised leaves treated with sucrose. Infecting the tissues with rice blast (Magnaporthe grisea) as a biotic stressor increased the expression of OsCIN1, OsCIN4, and OsCIN5, suggesting that these genes may participate in a switch in metabolism to resist pathogen invasion. These results demonstrate that OsCIN genes play diverse roles involving the regulation of metabolism, growth, development, and stress responses. PMID- 15759121 TI - [Comparison of diameter response of retinal arteries and veins to flickering light. A clinical study with healthy people]. AB - PURPOSE: The aim of this study was to compare the diameter response of retinal arteries and veins after provocation with flickering light regarding the amplitude and temporal course. METHODS: The study included 26 healthy volunteers. The vessel diameter was automatically and continuously measured by the retinal vessel analyzer. The examination consisted of a baseline measurement (100 s) followed by five periods of 20-s flicker light provocation and 80-s follow-up observation. RESULTS: A diameter dilation was observed immediately after the end of the flicker period in arteries (6.9+/-2.8%) and veins (6.5+/-2.8%, difference n.s.). The quotient of arterial and venous dilation (AVDQ) was 1.25+/-0.69 (range: 0.2-2.8). A significant correlation of age and arterial or venous dilation or the AVDQ was not observed. The arterial diameter reduced faster than the venous and reached a minimum of -2.7+/-1.4% at 25.9+/-10.8 s after the end of the flicker phase. The veins were dilated by 0.5+/-1.3% at the time of the maximal individual arterial constriction (p<0.0001). CONCLUSIONS: Flicker-evoked response of retinal arteries and veins did not differ in the dilative amplitude but in the temporal course of the abatement of the dilation. PMID- 15759122 TI - [Programmed cell death in the retina. Molecular mechanisms and therapeutic strategies]. AB - The cellular suicide program (apoptosis) is important not only for many physiological processes spanning from embryonal development to aging but also for the pathogenesis and the course of a wide spectrum of diseases. Among these disorders are chronic neurodegenerative diseases including acquired and inherited degeneration of photoreceptors as well as retinal ganglion cells during retinitis pigmentosa, macular degeneration, optic neuritis, or glaucoma. In the following review we will address general pro- and antiapoptotic signal transduction cascades, their relevance for retinal degeneration focussing on retinal ganglion cells, and new therapeutic strategies evolving from these findings. PMID- 15759123 TI - Intussusception in adults: an unusual and challenging condition for surgeons. AB - BACKGROUND AND AIMS: Intestinal intussusception in adults is a rare entity and there is an ongoing controversy regarding the optimal management of this problem. The purpose of this study was to determine the causes and management of intussusception in adults. PATIENTS AND METHODS: A retrospective review of patients more than 18 years of age with a diagnosis of intestinal intussusception between January 1996 and December 2003 was conducted. Data related to presentation, diagnosis, treatment, and pathology were analyzed. FINDINGS: A total of 13 patients were operated on due to intestinal intussusception. There were 6 men and 7 women with a mean age of 45 years (range 24--61 years). Abdominal pain was the most common presenting complaint (100%). Eight (61.5%) patients presented with acute symptoms and underwent emergency laparotomy. The diagnosis of gastrointestinal intussusception was made preoperatively only in 4 (30.7%) patients by abdominal ultrasonography and computerized tomography. The lead point of intussusception was located in the small intestine in 10 (76.9%) patients, in the colon in 2 (15.4%), and in the ileocecal valve in 1 (7.7%). A pathologic cause for the intussusception was identified in 12 (92.3%) cases and 1 (7.7%) was idiopathic. Of the cases with a defined cause, 58% of the cases were benign and 42% were malignant. Forty percent of cases of small bowel intussusception and 33.3% of cases of colonic intussusception were due to malignant lesions. All cases of small intestinal intussusception were reduced and no perforation occurred. Segmental intestinal resection was performed in 9 patients and excision of the Meckel's diverticulum was made in 1. In cases of colonic intussusception, reduction was not attempted and en-bloc resection was carried out. No perforation or spillage of the contents of the intussusception was observed. There was no surgical mortality. CONCLUSION: Adult intussusception is an unusual and challenging condition that represents a preoperative diagnostic difficulty. We think that colonic intussusceptions should be resected in an en bloc manner without reduction due to the risk of perforation and spillage of micro-organisms and malignant cells, whereas cases of small intestinal intussusception can be reduced without complications unless there is strangulation. PMID- 15759125 TI - Unexpected sudden death caused by medullary brain lesions involves all age groups and may include 'sudden infant death syndrome' as a subset. PMID- 15759124 TI - Lymph node micrometastasis in stage II distal rectal cancer following neoadjuvant chemoradiation therapy. AB - OBJECTIVE: The objective was to determine the presence and frequency of micrometastasis in lymph nodes of patients with rectal cancer treated by preoperative chemoradiation followed by curative resection. PATIENTS AND METHODS: All 56 patients included were treated with 5-FU and leucovorin plus 5,040 cGy, followed by radical surgery and were diagnosed with stage II distal rectal adenocarcinoma after complete pathological examination (ypT3-4N0M0). Immunohistochemistry was assessed with cytokeratin monoclonal antibody AE1/AE3. Three 4-microm paraffin sections were obtained from each lymph node, cut at 50 microm apart from each other. The results were reviewed by two independent pathologists. RESULTS: Mean number of lymph nodes was 9.6 per patient. Four patients (7%) and seven lymph nodes (1.35%) were positive for micrometastasis. Three patients had pT3 and one a pT4 tumor. One of the patients had positive micrometastasis and the presence of mucinous deposits. One other patient had mucinous deposits without any micrometastasis. All four patients are alive with no evidence of recurrent disease. Fourteen patients negative for micrometastasis had recurrent disease (25%), eight systemic (14.7%) and six locoregional (10.3%). There were two cancer-related deaths. The mean follow-up period was 39 months. CONCLUSION: Patients with rectal cancer treated by preoperative chemoradiation showed a surprisingly low rate of micrometastasis detection (7%), even in high risk patients (T3 and T4 tumors). Lymph node micrometastasis was not associated with decreased overall or disease-free survival. The identification of mucinous deposits on lymph nodes with no viable tumor cells may be direct evidence of lymph node downstaging. The downstaging effect of preoperative chemoradiation therapy may be significant in reducing even micrometastasis detection in low rectal cancer managed by this treatment strategy. PMID- 15759127 TI - Characteristics of neuronal lipofuscin in the superior temporal gyrus in Alzheimer's disease do not differ from non-diseased controls: a comparison with disease-related changes in the superior frontal gyrus. AB - Neuronal lipofuscin characteristics in the superior temporal gyrus from 21 patients with Alzheimer's disease (AD) and from 18 age-matched non-diseased subjects were compared with previously reported findings from the superior frontal gyrus. A discriminant function analysis of lipofuscin characteristics in the superior temporal gyrus did not provide a significant predictive level for cases whose diagnoses were correctly classified (56.4%, P=0.63). In contrast, AD related decrease in the number of smaller lipofuscin regions in the neurons of the frontal gyrus was confirmed, and the same analysis of lipofuscin characteristics in this region gave a significant predictive level for membership of the AD group of 86.6% (P<0.001). The findings indicate that changes in neuronal lipofuscin related to AD, which may reflect an increased rate of lipofuscin formation, show differences between neocortical regions. This study provides additional information on the distribution of neuropathological characteristics in AD. PMID- 15759126 TI - Thrombus and encapsulated hematoma in cerebral cavernous malformations. AB - Thrombi, encapsulated hematomas, and granulation tissue are frequently seen in cerebral cavernous malformations (CCMs). We investigated the role that these histological changes play in repeated hemorrhages in CCMs as well as lesion growth, examining specimens of CCMs surgically harvested from 20 patients. The immunohistochemical study included thrombomodulin (TM) and endothelial cell protein C receptor (EPCR), which are important regulators of blood coagulation. Thick capsules, which contained blood degradation product, were seen in cases with encapsulated hematomas. Clusters of sinusoidal vessels were found outside of these thick capsules. Granulation tissue with inflammatory infiltrates and capillaries was seen in 4 cases with non-capsulated hematomas. Organizing thrombi were seen in sinusoidal vessels in 15 out of 20 cases. Factor VIII-related antigen staining demonstrated numerous capillaries in and around organizing thrombi and within the thickened vessel walls as well as in both the inner and outer sides of the hematoma capsule. TM and EPCR were positive in the endothelial cells of these capillaries, whereas they were negative in those of capillaries in the brain surrounding the lesions. Our study suggests that thrombosed sinusoidal blood vessels could gradually expand by repeated bleeding from numerous capillaries inside the wall and become encapsulated hematomas, and capillaries outside the thickened vessel wall could become sinusoidal blood vessels. Thrombosis within cerebral venules could be one of the causal factors of CCMs. PMID- 15759128 TI - Silver staining profiles distinguish Pick bodies from neurofibrillary tangles of Alzheimer type: comparison between Gallyas and Campbell-Switzer methods. AB - Silver staining profiles of Pick bodies (PBs) and their relation to tau-like immunoreactivity were examined on hippocampal sections and compared with those of neurofibrillary tangles of Alzheimer type (NFTs). Pairs of mirror sections were double-fluorolabeled with an anti-paired helical filament tau (AT8) antibody and thiazin red (TR), a fluorochrome that identifies fibrillary structures such as NFTs. One of the paired sections was subsequently stained using the Gallyas method (GAL), and the other using the Campbell-Switzer method (CS). By comparison of the same microscopic field on fluorolabeled sections and on both silver stained paired sections, four different profiles of each structure could be distinguished: AT8 immunoreactivity, affinity to TR, argyrophilia with GAL or CS staining. PBs, containing mainly three-repeat (3R) tau, were positive for CS but not for GAL and its affinity to TR was, at most, weak. This selective affinity of PBs to CS is in sharp contrast with tau-positive structures of corticobasal degeneration/progressive supranuclear palsy, which are positive for GAL but not for CS, as we reported previously. This contrast is explainable if the argyrophilia with CS is related to deposits containing 3R tau, while that with GAL is linked to those containing four-repeat (4R) tau. Indeed, NFTs, containing both 3R and 4R tau, are positive for both CS and GAL, as expected. Taken together, differences in molecular composition of tau protein in these deposits are linked to their argyrophilic properties that are dependent on the staining method. Although explanations for these empirical differences are not yet available, awareness of this clear distinction is potentially of diagnostic and pathological relevance. PMID- 15759129 TI - Cytomegalovirus infection inhibits the expression of N-methyl-D-aspartate receptors in the developing mouse hippocampus and primary neuronal cultures. AB - Cytomegalovirus (CMV) is the most significant infectious cause of developmental brain disorders in humans. The infection occasionally persists and causes neurological disorders. The N-methyl-D-aspartate (NMDA) subtype of glutamate receptors is essential for the development and plasticity of synapses, but also is involved in neuronal excitotoxicity during viral infection. Here we investigated the effects of murine CMV (MCMV) infection on the expression of NMDA receptors in the hippocampal neurons of neonatal mice and primary neuronal cultures. Viral antigen was mostly found in hippocampal pyramidal neurons from the CA1 to CA3. Image analysis of immunohistochemistry demonstrated that the expression of NMDA receptor subunit 1 (NMDA-R1) protein in CA1 neurons of MCMV infected brain was reduced to 40% of that in uninfected brain. The signal of in situ hybridization for NMDA-R1 mRNA was also decreased in CA1 neurons of MCMV infected brain. In primary neuronal cultures, reduction of NMDA-R1 expression in MCMV-infected neurons was also detected by immunocytochemistry and Western blotting. These results suggest that reduction of NMDA receptor expression by MCMV infection may cause a decrease in the susceptibility of the neurons to excitotoxic cell death, and may be related to the establishment of viral persistence and functional disturbances in MCMV-infected neurons. PMID- 15759130 TI - Differentiation of classic medulloblastoma into metastatic large cell medulloblastoma with focal rhabdoid differentiation in the absence of posterior fossa recurrence. AB - A case of classic medulloblastoma that metastasized, despite the absence of local recurrence, to extraneural sites 7 years after treatment is reported. The metastases were, in contrast to the primary tumor, of large cell type and displayed abortive myogenic and, in one site, also rhabdoid differentiation. The primary tumor expressed microtubule-associated protein 1B and neuron-specific nuclear protein (NeuN), and was desmin negative. The metastases were also positive for microtubule-associated protein 1B and NeuN, although the expression of the latter marker was weak and/or focal in two of four metastases and absent in the rhabdoid metastasis. They were, in contrast with the primary tumor, all strongly positive for desmin. The hSNF5/INI1 was expressed in the nuclei of all cells of the primary tumor and the metastases, including the one with rhabdoid differentiation. Two metastases were studied by cytogenetics. The composite karyotype of a large cell metastasis was 45~46,XY,add(1)(p36.1),t(2;8)(p21;q24.1),add(3)(q25),t(9;15)(q22;q13),add(12)(p11 2), +1approximately2mar,inc[cp12]/46,XY[12], while the rhabdoid metastasis contained additional changes including monosomy 22. These findings indicate that some rhabdoid (atypical teratoid/rhabdoid) tumors of the cerebellum and medulloblastoma may be histogenetically related. PMID- 15759131 TI - Changes in hippocampal GABABR1 subunit expression in Alzheimer's patients: association with Braak staging. AB - Alterations in the gamma-aminobutyric acid (GABA) neurotransmitter and receptor systems may contribute to vulnerability of hippocampal pyramidal neurons in Alzheimer's disease (AD). The present study examined the immunohistochemical localization and distribution of GABA(B) receptor R1 protein (GBR1) in the hippocampus of 16 aged subjects with a range of neurofibrillary tangle (NFT) pathology as defined by Braak staging (I-VI). GBR1 immunoreactivity (IR) was localized to the soma and processes of hippocampal pyramidal cells and some non pyramidal interneurons. In control subjects (Braak I/II), the intensity of neuronal GBR1 immunostaining differed among hippocampal fields, being most prominent in the CA4 and CA3/2 fields, moderate in the CA1 field, and very light in the dentate gyrus. AD cases with moderate NFT pathology (Braak III/IV) were characterized by increased GBR1-IR, particularly in the CA4 and CA3/2 fields. In the CA1 field of the majority of AD cases, the numbers of GBR1-IR neurons were significantly reduced, despite the presence of Nissl-labeled neurons in this region. These data indicate that GBR1 expression changes with the progression of NFT in AD hippocampus. At the onset of hippocampal pathology, increased or stable expression of GBR1 could contribute to neuronal resistance to the disease process. Advanced hippocampal pathology appears to be associated with decreased neuronal GBR1 staining in the CA1 region, which precedes neuronal cell death. Thus, changes in hippocampal GBR1 may reflect alterations in the balance between excitatory and inhibitory neurotransmitter systems, which likely contributes to dysfunction of hippocampal circuitry in AD. PMID- 15759132 TI - A new murine model of giant proximal axonopathy. AB - The aromatic gamma-diketone 1,2-diacetylbenzene (1,2-DAB), the putative active metabolite of the organic solvent 1,2-diethylbenzene, forms blue-colored polymeric protein adducts and induces the formation of amyotrophic lateral sclerosis (ALS)-like giant, intraspinal neurofilamentous axonal swellings in Sprague Dawley rats. The pathogenetic mechanism of this neuropathy has yet to be understood. We assessed whether these pathological changes are also seen in the C57BL/6 mouse, the animal of choice for toxicogenomic studies. Mice were treated intraperitoneally with 30, 35, 50, or 70 mg/kg 1,2-DAB or its inactive isomer 1,3 DAB per day (or on alternate days) for up to 43 days. Animals treated with 30 or 35 mg/kg per day 1,2-DAB, but not with 1,3-DAB, developed muscle spasms and progressive weakness, most prominently in hind limbs. Light microscopy revealed swollen axons in spinal anterior horns and proximal ventral roots, and to a lesser extent in dorsal root ganglia of 1,2-DAB-treated animals. Ultrastructural examination of swollen axons revealed clumps of maloriented 10-nm neurofilaments. Sciatic nerves showed clustering of axonal microtubules and other organelles. These findings are qualitatively comparable to those reported in rats treated with 1,2-DAB and represent a suitable phenotype with which to explore molecular mechanisms of proximal, giant neurofilamentous axonopathy using proteomic and genomic technologies. PMID- 15759133 TI - The enlarging spectrum of desminopathies: new morphological findings, eastward geographic spread, novel exon 3 desmin mutation. AB - A 52-year-old man, who had developed distal muscle weakness in legs and arms, was found to have distal muscle atrophy as well as cardiac arrhythmia. His 10-year younger brother developed restrictive cardiomyopathy at the age of 20 years, which required cardiac transplantation at the age of 41 years. Skeletal muscle biopsy specimens of the older brother revealed granulofilamentous material and plaques containing numerous proteins, foremost desmin, as did cardiac biopsy tissue. The explanted heart of the younger brother showed similar protein-rich plaques and granulofilamentous material within cardiac myocytes. A novel heterozygous Glu245Asp (E245D) missense mutation in exon 3 of the desmin gene (DES) at 2q35 was found in the older brother. While clinical data and muscle biopsy pathology of the older brother conform to the nosological spectrum of desminopathies, the early-onset cardiomyopathy, a similar cardiac pathology as in skeletal muscle tissues and a novel missense mutation in the DES gene, enlarge the nosological spectrum of desminopathies. PMID- 15759134 TI - Humanin detected in skeletal muscles of MELAS patients: a possible new therapeutic agent. AB - Humanin (HN) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease related insults. We have previously indicated that HN increases cellular ATP levels and speculated that this peptide may rescue energy-deficient cells in mitochondrial disorders. Here, we report, for the first time, increased HN expression in skeletal muscles from patients with mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS). HN was strongly positive in all ragged-red fibers (RRFs) and some non-RRFs, and most of them were type 1 fibers generally requiring higher energy than type 2 fibers. HN in these fibers was localized in mitochondria. HN expression was also increased in small arteries that strongly reacted for succinate dehydrogenase. Our experiments on muscular TE671 cells indicated the possibility that synthesized HN increases cellular ATP levels by directly acting on mitochondria. From these in vivo and in vitro findings, we propose that HN expression might be induced in response to the energy crisis within affected fibers and vessels in MELAS muscles and further be a possible therapeutic candidate for MELAS. PMID- 15759135 TI - Photodynamic therapy using a novel photosensitizer, ATX-S10(Na): comparative effect with 5-aminolevulinic acid on squamous cell carcinoma cell line, SCC15, ultraviolet B-induced skin tumor, and phorbol ester-induced hyperproliferative skin. AB - Photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) is available for the treatment of actinic keratosis (AK). Recently, we developed a new PDT photosensitizer, ATX-S10(Na), and have shown that ATX-S10(Na) PDT is effective for the treatment of various human skin diseases, such as squamous cell carcinoma, Bowen's disease, basal cell carcinoma, and psoriasis. In the present study, we compared the effects of ATX-S10(Na) PDT and ALA PDT on hyperproliferative skin induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), on the squamous cell carcinoma cell line, SCC15, in vitro, and on UVB-induced skin tumors in vivo. TPA treatment induced epidermal acanthosis, which was more markedly suppressed by ATX-S10(Na) PDT than by ALA PDT. ATX-S10(Na) PDT more effectively eliminated UVB-induced AK and squamous cell carcinoma (SCC) than ALA PDT. Furthermore, both ATX-S10(Na) PDT and ALA PDT induced the death of SCC15 cells, and the effect of ATX-S10(Na) PDT was greater than that of ALA PDT. Our results indicate that ATX-S10(Na) PDT might be more effective than ALA PDT for the treatment of various skin diseases. PMID- 15759137 TI - Use of multivariate curve resolution for determination of chromium in tanning samples using sequential injection analysis. AB - We report a method for determining total chromium in tanning samples using sequential injection analysis (SIA) with a diode-array spectrophotometric detector. With a suitable analytical sequence CrO4(2-) is converted to Cr2O7(2-) inside the tubes of the SIA system, after total oxidation of chromium(III). A data matrix is obtained and analysed by several chemometric techniques based on multivariate analysis: principal components analysis, simple-to-use interactive self-modelling mixture analysis, and multivariate curve resolution-alternating least-squares. We studied several samples from different stages of a tanning process. Two of these samples were easily oxidized but the others needed more extreme conditions. The analytical sequence prepared, which was based on obtaining a pH gradient and used H2SO4 as reagent, is valid and independent of the level of oxidation needed for the sample. We established a calibration model and evaluated the figures of merit. In some samples we found interferents. With this method the amounts of chromium in each sample were quantified and the results were statistically similar to those obtained by use of the reference method, atomic absorption spectrometry. PMID- 15759136 TI - Is bortezomib, a proteasome inhibitor, effective in treating cancer-associated weight loss? Preliminary results from the North Central Cancer Treatment Group. AB - BACKGROUND: Weight loss predicts a poor prognosis for cancer patients, and previous studies have implicated the ubiquitin-proteasome pathway as a major mediator of cancer-associated weight loss. The recent emergence of bortezomib, a proteasome inhibitor, now allows testing on whether proteasome inhibition is effective therapy for cancer-associated weight loss. METHODS: This study represents a subanalysis from two prior antineoplastic trials in patients with adenocarcinoma of the pancreas. The first included 46 patients with metastatic pancreatic cancer who were treated with single-agent bortezomib (intravenous doses of 1.5 or 1.3 mg/m2 on days 1, 4, 8, and 11 of a 21-day cycle). The second included 42 patients with pancreatic cancer treated with single-agent octreotide (200 or 500 microg subcutaneously three times a day). The FACT-C questionnaire provided appetite and related data for bortezomib-treated patients. Serial weight data were available from both trials. Such data from the octreotide trial were utilized for comparative purposes because the latter holds no track record in treating cancer-associated weight loss. RESULTS: Bortezomib- and octreotide treated patients were roughly comparable at baseline, and neither agent demonstrated notable antineoplastic effects. FACT-C data suggested stable appetite, but high patient dropout rates invite caution in interpretation. For example, in response to "I have a good appetite," mean scores for bortezomib treated patients were 45 at baseline (n=42), 45 at the end of cycle 1 (n=26), and 44 at the end of cycle 2 (n=9). In contrast, weight data appeared more straightforward to interpret: direct comparisons of mean change in weight from baseline between bortezomib- and octreotide-treated patients showed no significant differences between groups. CONCLUSIONS: These preliminary results suggest that bortezomib shows negligible favorable effects on cancer-associated weight loss in patients with metastatic pancreatic cancer. We conclude that further study of bortezomib specifically in this setting and for this indication is not warranted. PMID- 15759138 TI - Determination of free copper concentrations in natural waters by using supported liquid membrane extraction under equilibrium conditions. AB - A method is described for measurement of freely dissolved copper concentrations in natural water samples using supported liquid membrane (SLM) extraction under equilibrium conditions, a technique denoted "equilibrium sampling through membranes" (ESTM). For this purpose, 1,10-dibenzyl-1,10-diaza-18-crown-6 as neutral carrier and oleic acid were used in the membrane phase. The main variables optimised were the carrier used to form the metal complexes, the organic solvent used in the membrane, the countercation, pH, the ligand used in the acceptor phase, the extraction time, and the flow rate of the donor phase. After the optimisation process an enrichment factor of 18.5 was obtained. Equilibrium conditions were reached after extraction for 60 min if a flow rate of 1.0 mL min(-1) or greater was used. When different ligands such as humic acids, phthalic acid, and EDTA were added to the sample solution, and sample pH ranged from 6 to 8, the results obtained for freely dissolved copper concentrations were in a good agreement with results from speciation calculations performed with Visual Minteq V 2.30, Cheaqs V L20.1, and WinHumic V. The developed technique was applied to analysis of stream and leachate water. PMID- 15759139 TI - HPLC determination of residual monomers released from heat-cured acrylic resins. AB - HPLC was used to examine the leachability of three non-phthalic and four phthalic post-polymerized residual monomers from three commercially available heat-cured acrylic resins. Specimens of equal dimensions were constructed from each brand of material by following the standardized procedure and were stored under three different conditions, namely, distilled water, artificial saliva, and a binary mixture of ethanol/water. The resulting liquids provided samples for analysis by HPLC. Three different experiments were performed for each brand of acrylic and each storage condition in order to examine the effects of parameters, particularly time and temperature. The results obtained from this study suggest that a wide spectrum of residues diffuse out of the three examined acrylic resin materials. The non-phthalic compounds were leached at high concentrations, whereas all the phthalates examined exhibited different degrees of elusion commensurate with the storage condition, brand of material, and type of experiment. It seems that a significant quantity of non-phthalic and phthalic residues diffuse out of the acrylic resin materials examined. The main component extracted was methyl methacrylate, the level of which seems to be time-dependent and decreases for a period of up to 5 days when resins are stored in distilled water at room temperature. PMID- 15759140 TI - The effect of cooling on the acetylcholine-induced current of identified Helix pomatia Br neuron. AB - The Br neuron of the snail Helix pomatia, involved in neuronal regulation of various homeostatic and adaptive mechanisms, represents an interesting model for studying effects of temperature changes on neuronal activity of poikilotherms. The acetylcholine (ACh) induces a transient, inward dose-dependent current in the identified Br neuron. In the work presented, we analyses the effects of cooling on the ACh-induced inward current. The amplitude of ACh-induced inward current was markedly decreased after cooling and the speed of the decay of ACh response was decreased. Sensitivity to cooling of Ach-activated current on the Br neuron is mediated by a mechanism that does not involve change in the apparent receptor affinity or the cooperativity of binding. PMID- 15759141 TI - Expectations and outcomes: decision-making in the primate brain. AB - Success in a constantly changing environment requires that decision-making strategies be updated as reward contingencies change. How this is accomplished by the nervous system has, until recently, remained a profound mystery. New studies coupling economic theory with neurophysiological techniques have revealed the explicit representation of behavioral value. Specifically, when fluid reinforcement is paired with visually-guided eye movements, neurons in parietal cortex, prefrontal cortex, the basal ganglia, and superior colliculus-all nodes in a network linking visual stimulation with the generation of oculomotor behavior-encode the expected value of targets lying within their response fields. Other brain areas have been implicated in the processing of reward-related information in the abstract: midbrain dopaminergic neurons, for instance, signal an error in reward prediction. Still other brain areas link information about reward to the selection and performance of specific actions in order for behavior to adapt to changing environmental exigencies. Neurons in posterior cingulate cortex have been shown to carry signals related to both reward outcomes and oculomotor behavior, suggesting that they participate in updating estimates of orienting value. PMID- 15759142 TI - Real-time quantitative PCR assay for the detection of Helicobacter pylori: no association with sudden infant death syndrome. AB - Helicobacter pylori infection is known to be one of the most common chronic infectious diseases in humans. Recently, a hypothesis was proposed that H. pylori infection could be a frequent cause for sudden infant death syndrome (SIDS). We have investigated this postulated association by examining formalin-fixed paraffin-embedded gastric tissues of a retrospective cohort of 94 SIDS cases: The presence of H. pylori was inferred from a newly developed real-time quantitative PCR assay with SYBR Green I detection. This assay is based on the amplification of the single-copy H. pylori-specific glmM gene. Accuracy and precision were verified using a plasmid containing a 977-bp fragment of this glmM gene. The assay was very sensitive, and as few as 30 template copies per PCR reaction could be detected even in the presence of excess human DNA. The assay was validated on mucosal biopsy samples of patients with known H. pylori infections. Interfering effects due to SIDS gastric tissue were excluded. Only two (2.1%) of the SIDS samples yielded H. pylori DNA copy numbers and only beyond the lowest standard concentration. These results could be confirmed independently by immunohistochemistry using an H. pylori-specific antibody. Thus, an infection by H. pylori is very rare in cases of SIDS, and thus the postulated association of H. pylori infection and SIDS cannot be confirmed. PMID- 15759143 TI - Acetylcholinesterase in Hirschsprung's disease. AB - The association between the congenital absence of colonic ganglion cells and an increased acetylcholinesterase (AChE) expression in the affected tissue is of diagnostic importance in Hirschsprung's disease (HSCR). Investigation of AChE's function in development may also help unravel some of the complex pathophysiology in HSCR. Normal nerves do not stain for AChE, but increased AChE expression is associated with the hypertrophied extrinsic nerve fibres of the aganglionic segment in HSCR. Although a high degree of histochemical diagnostic accuracy exists, results are not always uniform, and false positives and false negatives are reported. False negative results are primarily related to age, and an absence of AChE reaction does not exclude HSCR in neonates within the first 3 weeks after birth. AChE staining results may lack uniformity, resulting in a number of technical modifications that have been made to improve standardization of AChE staining. At least two distinct histological patterns are described, types A and B. The interpretation of increased AChE staining patterns in ganglionated bowel at the time of surgical pull-through remains a problem in patients with HSCR. The development of rapid staining techniques has helped to identify normal ganglionated bowel with greater certainty. The presence of fine AChE neurofibrils in the ganglionated segment has contributed to the debate surrounding intestinal neuronal dysplasia. Quantitative assay of cholinesterase activity confirms the pattern of histochemical staining. AChE is particularly increased in relation to butrylcholinesterase, with one molecular form, the G4 tetrameric form, predominating. It is likely that the raised levels of AChE in aganglionic tissue are the transcriptional consequence of the abnormalities in signalling molecules that characterize HSCR. Evidence suggests that this AChE is functioning in a nonenzymatic capacity to promote cell adhesion and differentiation and that the hypertrophied nerves and neurofibrils may be the result of this increased AChE expression. PMID- 15759144 TI - Molecular models of protein targets from Mycobacterium tuberculosis. AB - Structural characterization of enzymes that belong to microbial metabolic pathways is very important for structure-based drug design since some of these proteins may be present in the bacterial genome, but absent in humans. Thus, metabolic pathways became potential targets for drug design. The motivation of this work is the fact that Mycobacterium tuberculosis is the cause of the deaths of millions of people in the world, so that the structural characterization of protein targets to propose new drugs has become essential. DBMODELING is a relational database, created to highlight the importance of methods of molecular modeling applied to the Mycobacterium tuberculosis genome with the aim of proposing protein-ligand docking analysis. There are currently more than 300 models for proteins from Mycobacterium tuberculosis genome in the database. The database contains a detailed description of the reaction catalyzed by each enzyme and their atomic coordinates. Information about structures, a tool for animated gif image, a table with a specification of the metabolic pathway, modeled protein, inputs used in modeling, and analysis methods used in this project are available in the database for download. The search tool can be used for researchers to find specific pathways or enzymes. PMID- 15759145 TI - Toxicity of nitrite toward mesophilic and thermophilic sulphate-reducing, methanogenic and syntrophic populations in anaerobic sludge. AB - The various problems associated with treating sulphate-containing wastewaters stem inherently from successful competitive interactions between sulphate reducing bacteria (SRB) and other bacteria involved in the process, resulting in the formation of H(2)S. Prevention of in-reactor sulphide generation by use of specific SRB inhibitors presents a potential solution. Nitrite has been reported to be a specific inhibitor of SRB but its possible toxicity to syntrophic and methanogenic members of the anaerobic consortium has not been investigated. In batch activity and toxicity tests, under both mesophilic and thermophilic conditions, nitrite, at concentrations of up to 150 mg L(-1), was found to be ineffective as a specific inhibitor of SRB, and was also shown to have an inhibitory effect on the activity of syntrophic and methane-producing bacteria in mesophilic and thermophilic digester sludge samples. PMID- 15759146 TI - Apoptosis induced by oxalate in human renal tubular epithelial HK-2 cells. AB - Oxalate is not only considered to be one of the main constituents of urinary stones, but it also has toxic effects on renal tubular epithelial cells, affecting the pathogenesis of nephrolithiasis. We tried to elucidate the effects of oxalate on human renal tubular epithelial cells (HK-2 cells). In addition, we investigated whether the toxic effect of oxalate occurs by apoptosis, and determined the expression of Bcl-2 family and caspase 9 proteins known as apoptosis-related protein. HK-2 cells were incubated with different concentrations of oxalate, and the effect of oxalate on the growth of the cells was assessed by MTT assay. A caspase-3 activity assay and TUNEL assay were performed on HK-2 cells after oxalate exposure in order to evaluate apoptosis. Immunoblot analysis of Bax, Bcl-2, Bcl-xL, and caspase-9 were performed. Oxalate exposure resulted in significant growth inhibition of HK-2 cells as oxalate concentrations increased. The toxic effect of oxalate on HK-2 cells was considered to occur through apoptosis, as suggested by the increase of caspase-3 activity. The percentage of positive nuclei stained using the TUNEL method was 18+/-2.3 in oxalate-treated cells and 2.5+/-0.9 in untreated cells (P<0.05). Bax and caspase-9 protein expression increased significantly as oxalate concentrations increased, but Bcl-2 protein expression decreased. There was no difference in Bcl-xL protein expression among the various concentrations of oxalate. Our results show that oxalate has a toxic effect on HK-2 cells and that this effect is induced by apoptosis, which may be mediated by an intrinsic pathway. PMID- 15759147 TI - The route of administration (oral vs intravenous) does not influence dose or outcome in Graves' disease and unifocal autonomy. AB - PURPOSE: In a prospective randomised study, we investigated the influence of the route of administration of radioiodide on dosimetry and therapy outcome. METHODS: Fifty-four patients suffering from Graves' disease (GD) and 60 patients with unifocal autonomy (UA) participated in the study and were randomly treated with either orally or intravenously administered radioiodide. Pretherapeutic dosimetry was based on single uptake measurements with a calibrated uptake probe system. The radioiodine kinetics during hospitalisation was assessed by daily bedside uptake measurements. Therapeutic dose was determined by half-life and thyroid uptake at the time of discharge using the same uptake probe as for the radioiodine test. RESULTS: No improvement in accuracy of dosimetry was achieved when radioiodide was administered intravenously. Mean therapeutic doses were identical following intravenous or oral administration. Variation in the achieved dose was slightly higher in the patients receiving oral administration, this being attributable to larger deviations in discrete activities of the capsules administered as compared with the values determined by dosimetry. No differences according to treatment modality were found with regard to therapeutic outcome. Eighty-seven patients attended 6-month follow-up after therapy. In the UA group, successful treatment, defined as a normal or elevated TSH level, was observed in 94% of patients after oral administration and in 80% after intravenous administration; corresponding figures in the GD group were 68% and 65%. CONCLUSION: The causes of individual differences between targeted and therapeutically achieved doses remain undetermined. Variations in the bioavailability of radioiodide or other parameters affecting thyroid status may be involved, and further investigations are needed to clarify this. PMID- 15759148 TI - Brain FDG PET study of normal aging in Japanese: effect of atrophy correction. AB - PURPOSE: The aim of this study was to investigate the effects of atrophy correction on the results of 18F-fluorodeoxyglucose positron emission tomography (FDG PET) in the context of normal aging. METHODS: Before the human study was performed, a Hoffman 3D brain phantom experiment was carried out in order to validate a newly developed correction method for partial volume effects (PVEs). Brain FDG PET was then performed in 139 healthy Japanese volunteers (71 men, 68 women; age 24-81 years). PET images were corrected for PVEs using grey matter volume, which was segmented from co-registered magnetic resonance images and convoluted with the spatial resolution of the PET scanner. We investigated the correlation between advancing age and relative regional FDG activity, which was normalised to the global activity before and after PVE correction using Statistical Parametric Mapping 99. RESULTS: The PET image, when corrected for PVEs, provided more homogeneous tracer distribution in the whole phantom than in the original PET image. The human PET study of both sexes revealed significant negative correlations between age and relative FDG activity in the bilateral perisylvian and medial frontal areas before PVE correction. However, these negative correlations were largely resolved after PVE correction. CONCLUSION: Correction for PVEs was effective in our FDG PET study. The reduction in FDG uptake with advancing age that was detected by FDG PET without PVE correction could be accounted for largely by an age-related cerebral volume loss in the bilateral perisylvian and medial frontal areas. PMID- 15759150 TI - The sentinel node in cervical cancer: scintigraphy and laparoscopic gamma probe guided biopsy. PMID- 15759151 TI - Modulators of the glycine site on NMDA receptors, D-serine and ALX 5407, display similar beneficial effects to clozapine in mouse models of schizophrenia. AB - RATIONALE: Schizophrenia is characterized by disturbances in sensorimotor gating and attentional processes, which can be measured by prepulse inhibition (PPI) and latent inhibition (LI), respectively. Research has implicated dysfunction of neurotransmission at the NMDA-type glutamate receptor in this disorder. OBJECTIVES: This study was conducted to examine whether compounds that enhance NMDA receptor (NMDAR) activity via glycine B site, D-serine and ALX 5407 (glycine transporter type 1 inhibitor), alter PPI and LI in the presence or absence of an NMDAR antagonist, MK-801. METHODS: C57BL/6J mice were tested in a standard PPI paradigm with three prepulse intensities. LI was measured in a conditioned emotional response procedure by comparing suppression of drinking in response to a noise in mice that previously received 0 (non-preexposed) or 40 noise exposures (preexposed) followed by two or four noise-foot shock pairings. RESULTS: Clozapine (3 mg/kg) and D-serine (600 mg/kg), but not ALX 5407, facilitated PPI. MK-801 dose dependently reduced PPI. The PPI disruptive effect of MK-801 (1 mg/kg) could be reversed by clozapine and ALX 5407, but not by D-serine. All the compounds were able to potentiate LI under conditions that disrupted LI in controls. MK-801 induced abnormal persistence of LI at a dose of 0.15 mg/kg. Clozapine, D-serine, and ALX 5407 were equally able to reverse persistent LI induced by MK-801. CONCLUSIONS: D-Serine and ALX 5407 display similar effects to clozapine in PPI and LI mouse models, suggesting potential neuroleptic action. Moreover, the finding that agonists of NMDARs and clozapine can restore disrupted LI and disrupt persistent LI may point to a unique ability of the NMDA system to regulate negative and positive symptoms of schizophrenia. PMID- 15759152 TI - Evidence for improved performance in cognitive tasks following selective NR2B NMDA receptor antagonist pre-treatment in the rat. AB - RATIONALE: We previously reported that the NR2B subunit-selective N-methyl-D aspartate (NMDA) antagonist Ro 63-1908 produced a marked deficit in response control in the five-choice serial reaction time task (5-CSRTT). OBJECTIVES: The present studies were designed to investigate this further by studying the NR2B NMDA antagonists, ifenprodil, traxoprodil (CP101,606), Ro 25-6981 as well as Ro 63-1908 in this test. METHODS: Following training in the 5-CSRTT, separate groups of rats were either tested under (1) standard test conditions [5 s inter-trial interval (ITI), 0.5 s stimulus duration, 100 trials], (2) high (3 s ITI) and low (10 s ITI) event rate of stimulus presentation and (3) a 250-trial protocol in aged 2-year-old rats. In a final study, the effects of traxoprodil were investigated in an operant delayed match to position (DMTP) task, a test of working memory, and compared to dizocilpine and Ro 63-1908. RESULTS: Similar to Ro 63-1908, both traxoprodil (1-10 mg/kg) and Ro 25-6981 (3--30 mg/kg) increased premature responding but also increased response speed with no error trade-off. Conversely, ifenprodil (1--10 mg/kg) slowed response speed and increased omissions with no effect on premature responding. Tested under a variable ITI, Ro 63--1908 (1 mg/kg) increased premature responding at all ITIs, but this change was proportional to controls. At short ITI (3 s), Ro 63-1908 reliably improved performance both in terms of response speed and accuracy (percent correct). In a 250-trial protocol in aged rats, both Ro 63-1908 (0.1-0.3 mg/kg) and, particularly, traxoprodil (1--3 mg/kg) improved performance-increasing response speed and increasing the number of rewards earned during test. Finally, traxoprodil (1--10 mg/kg) improved accuracy and increased response speed in the DMTP task. CONCLUSIONS: The present studies support the view that selective NR2B NMDA antagonists promote impulsive-type responding in the 5-CSRTT; however, under certain test conditions, drugs of this class-notably traxoprodil-may also improve task performance. PMID- 15759153 TI - Phylogenomic analysis and expression patterns of large Maf genes in Xenopus tropicalis provide new insights into the functional evolution of the gene family in osteichthyans. AB - We have performed an exhaustive characterization of the large Maf family of basic leucine zipper transcription factors in vertebrates using the genome data available, and studied the embryonic expression patterns of the four paralogous genes thus identified in Xenopus tropicalis. Our phylogenetic analysis shows that, in osteichthyans, the large Maf family contains four orthology classes, MafA, MafB, c-Maf and Nrl, which have emerged in vertebrates prior to the split between actinopterygians and sarcopterygians. It leads to the unambiguous assignment of the Xenopus laevis XLmaf gene, previously considered a MafA orthologue, to the Nrl class, the identification of the amphibian MafA and c-Maf orthologues and the identification of the zebrafish Nrl gene. The four X. tropicalis paralogues display partially redundant but nevertheless distinct expression patterns in the somites, developing hindbrain, pronephros, ventral blood island and lens. Comparisons with the data available in the mouse, chick and zebrafish show that these large Maf expression territories are highly conserved among osteichthyans but also highlight a number of differences in the timing of large Maf gene expression, the precise extent of some labelled territories and the combinations of paralogues transcribed in some organs. In particular, the availability of robust phylogenies leads to a reinterpretation of previous expression pattern comparisons, suggesting an important part for function shuffling within the gene family in the developing lens. These data highlight the importance of exhaustive characterizations of gene families for comparative analyses of the genetic mechanisms, which control developmental processes in vertebrates. PMID- 15759154 TI - Experimental schistosomiasis, protective aspects of granulomatous reaction in the mouse liver. AB - We show two mechanisms of liver protection by the granulomatous reaction against Schistosoma mansoni eggs entrapped in the organ. First, eosinophil peroxidase and its substrate H(2)O(2) are released by inflammatory cells in the immediate vicinity of the parasite eggs. The efficiency of this process was demonstrated by administration of antioxidants to infected mice. The treatment, which reduces H(2)O(2) production, significantly improved the ability of parasite eggs to hatch after collection from the liver. Secondly, we labeled the released egg antigens in liver histological sections and we found that the lattice of collagen fibers which is built around eggs appears to create a barrier preventing released compounds from diffusing freely in surrounding tissues. Together, oxidative processes and antigen containment allow the parasitized liver to cope with the dual threat posed by parasite eggs, i.e. a highly resistant chitinous eggshell and the release of toxic substances. PMID- 15759155 TI - Apoptosis patterns in experimental Taenia solium and Taenia crassiceps strobilae from golden hamsters. AB - Apoptosis or programmed cell death (PCD) patterns of two taeniid species, Taenia solium and Taenia crassiceps, were explored in adult tapeworms grown in golden hamsters. Animals were fed either ten viable T. solium cysticerci from naturally infected pigs or from T. crassiceps WFU strain maintained in Balb/c mice. Adult strobilae were recovered from the intestine at different times after infection and either frozen at -70 degrees C or fixed in paraformaldehyde-glutaraldehyde. Frozen sections were processed using the DNA fragmentation fluorescent TUNEL reagents and examined in an epifluorescent microscope. Fixed tissues were processed for light and electron microscopy. Typical apoptotic cells were found in the central core of scolex and strobilar tissues, mainly in the germinal tissue and subtegumentary areas. By the TUNEL technique, cells exhibited the characteristic fluorescent images of condensed nuclear chromatin. By light microscopy of thick sections stained with toluidine blue, we found a number of small rounded cells which had lost their cytoplasmic bridges and had shrunken nuclei with aggregated chromatin, cells which were found interspersed with normal syncytial cells. Similar cell morphology was confirmed by electron microscopy. Stunted viable worms, recovered with longer mature specimens, had very short strobilae and exhibited a large number of apoptotic cells in the germinal neck tissues. The results are consistent with the syncytial nature of these parasites, and strongly suggest that cell proliferation and PCD in these adult cestodes are continuous processes of the germinal tissue and tegumentary cytons. PMID- 15759156 TI - IgG isotype to C-terminal 19 kDa of Plasmodium vivax merozoite surface protein 1 among subjects with different levels of exposure to malaria in Brazil. AB - Subclasses of antibodies to the C-terminal 19 kDa fragment of the Plasmodium vivax merozoite surface protein 1 (PvMSP-1(19)) were assessed among subjects with distinct degrees of malaria exposure in the Brazilian endemic area. The PvMSP 1(19) specific IgG1and IgG3 levels were low among subjects with long-term exposure (approximately 19 years) when compared to subjects less and sporadically exposed (<1 year). No statistically difference was observed in IgG subclass distribution of antibodies from symptomatic Plasmodium-infected patients, asymptomatic parasite carriers and non-infected subjects living in a same mesoendemic area. Subjects briefly exposed to a P. vivax outbreak living in a rural community outside the endemic area were also evaluated to measure the persistence of specific antibodies. IgG anti-PvMSP-1(19) antibodies persisted in 40% of the subjects who had had malarial symptoms 8 months before and decreased after 7 years (28%). Specific IgG1 were the predominant isotype. Our study emphasizes the highly immunogenicity of the PvMSP-1(19) and points toward its possible use as a potential malaria vaccine. PMID- 15759157 TI - Dexamethasone treatment induces susceptibility of outbred Webster mice to experimental infection with Besnoitia darlingi isolated from opossums (Didelphis virginiana). AB - The Sarcocystidae comprise a diverse, monophyletic apicomplexan parasite family, most of whose members form intracellular cysts in their intermediate hosts. The extent of pathology associated with such cyst formation can range widely. We currently lack experimental animal models for many of these infections. Here we explored dexamethasone treatment as a means to render outbred mice susceptible to Besnoitia darlingi infection and demonstrated that this approach allows viable parasites to be subsequently isolated from these mice and maintained in tissue culture. Besnoitia bradyzoites recovered from crushed cysts derived from naturally infected opossums (Didelphis virginiana) replicated and reproduced the development of besnoitiosis in mice treated with dexamethasone (0.5 mg/ml drinking water) daily for 12 days post infection (DPI). Isolates recovered from the peritoneal exudates of these mice were viable and were maintained in long term tissue cultures. In contrast, control mice given saline without dexamethasone and challenged with similar bradyzoites remained clinically normal for up to 70 DPI. An additional group of mice challenged with the same inoculum of bradyzoites and given dexamethasone at the same concentration and treated with sulfadiazine (1 mg/ml drinking water) daily for 12 DPI also remained normal for up to 70 DPI. Severe disease developed more rapidly in dexamethasone-treated mice inoculated with culture-derived B. darlingi tachyzoites than in those inoculated with cyst-derived bradyzoites. B. darlingi tachyzoite-infected, untreated control mice developed signs of illness at 18 DPI. In contrast, mice treated with sulfadiazine showed no clinical signs up to 50 DPI. Although dexamethasone treatment was required to establish B. darlingi infection in outbred mice inoculated with opossum-derived B. darlingi bradyzoites, no such treatment was required for mice inoculated with culture-derived B. darlingi tachyzoites. Finally, sulfadiazine was highly effective in protecting mice from infection with the tachyzoite stage of B. darlingi. PMID- 15759158 TI - Monstrous craniopharyngioma. Case presentations and term proposal. AB - INTRODUCTION: Craniopharyngiomas (CF) are benign tumors, which can be cured by total resection; however, this is not always possible to achieve, thus leading to tumor recurrence. When these tumors achieve disproportionate growth, the treatment is even more difficult, fortunately grotesque CF are not frequent, making experience and data collection more arduous to obtain. CASE REPORTS: Four patients are presented here to illustrate the need for the use of a new term, "monstrous craniopharyngioma," which is proposed with the aim of making the evaluation of the different kinds of treatment available more accurate. CONCLUSIONS: Craniopharyngiomas that have grown into more than one cranial fossa with mixed solid and cystic components pose a special challenge for resection, and therefore a systematic classification and approach are required in order to obtain the best surgical results. PMID- 15759159 TI - Serum growth hormone and insulin but not insulin-like growth factor-1 levels are elevated in patients with fibromyalgia syndrome. AB - Standard radioimmunoassay (RIA) was employed to quantify basal serum growth hormone (GH), insulin-like growth factor-I (IGF-1), and insulin levels in 32 normoglycemic patients with clinically active fibromyalgia and in 29 normoglycemic control subjects. The GH concentration was significantly higher (P < 0.001) in female fibromyalgia patients than age-matched, normal female subjects. In contrast, basal serum IGF-1 concentrations did not differ between these groups. A scatter plot generated from two-stage, least-squares analysis showed that serum GH lacked correlation with the serum IGF-1 concentrations of normal female subjects (P = 0.73) and female fibromyalgia patients (P = 0.19). In addition to the results from serum GH and IGF-1 RIA, we also found significantly higher fasting serum insulin levels (P = 0.03) in male fibromyalgia patients and a trend toward elevated fasting serum insulin levels in the female fibromyalgia population ( P = 0.07), with the mean fasting level in the male fibromyalgia group (35.7 microU/ml(-1)) exceeding the upper limit of normal serum insulin levels (i.e., 27 microU/ml(-1)). Based on these results, basal serum GH and fasting serum insulin levels appear to be valuable surrogate markers in clinically active, normoglycemic fibromyalgia patients. PMID- 15759160 TI - [Inpatient treatment of depression. Should one combine psychotherapy and drugs?]. AB - Antidepressants as well as different psychotherapeutic strategies have been proven efficacious in the treatment of unipolar depression. In the clinical setting both are often combined using psychotherapeutic methods varying from psychoeducation to formal psychotherapy. The present article provides a critical overview of the evidence base for this combination in the inpatient treatment of depression. The current literature is contradictory and difficult to compare. However, combination therapy appears advantageous in therapy-resistant, chronic and severe forms of depressive disorders. Much further research is needed to facilitate well-founded guidelines. PMID- 15759161 TI - [Guideline conformity and outcome of inpatient treatment for schizophrenia. A clinical comparison]. AB - Patient outcome and guideline conformity in inpatient schizophrenia treatment was systematically evaluated and compared with 597 patients across seven psychiatric hospitals. Patient structure and treatment processes showed a great variability between hospitals. Patient characteristics, especially mental state, and the chronicity of the disease were the strongest predictors of clinical outcome. Outcome evaluation using quality indicators is only possible after case-mix adjustment taking into account prognostic factors. A poorer average clinical outcome was associated with lower guideline conformity in a variety of treatment domains. After case-mix adjustment, benchmarking is an opportunity to improve quality of treatment and promote guideline conformity. PMID- 15759162 TI - [Generalized compartment syndrome after excessive drinking. A rare complication of psychological disorders?]. AB - Psychogenic polydipsia can lead to compartment syndromes, which is too infrequently considered in psychiatric patients who binge-drink on hypotonic fluids. If masked by the leading clinical presentation of cerebral edema, compartment syndromes of the extremities may be diagnosed too late or remain undetected. Based on a literature review and case report, we discuss additional factors and the specific features of diagnosis and treatment. PMID- 15759163 TI - [Psychiatrists as victims of stalking]. AB - Stalking is defined as notorious following, menacing, harassing, and contacting of a victim against his or her declared will. Such behavior can occur in a variety of psychiatric disorders. Attending physicians can also become victims of stalking. Especially psychiatrists appear to have a high risk of being stalked. Up to now, only a few studies investigated stalking behavior of patients aimed at medical professionals. In most cases, it was investigated together with aggressive behavior and sexual harassment. Cases of psychiatrists being stalked by patients or clients have not been described in detail. This case report describes a patient with erotomania who developed stalking behavior and victimized his attending psychiatrist. The relevance of stalking for clinical practice, the context of stalking behavior, and underlying psychiatric disorders and legal issues are discussed. PMID- 15759164 TI - [Unusual course of alpha-mannosidosis with symptoms of paranoid-hallucinatory psychosis]. AB - We report the case of a 27-year-old female with recurrent paranoid-hallucinatory episodes who was initially diagnosed as suffering from schizophrenic psychosis. After 10 years of treatment under this diagnosis, alpha-mannosidosis was identified to be the underlying cause of her psychiatric symptoms. alpha Mannosidosis is a rare autosomal recessive lysosomal storage disorder associated with decreased activity of the enzyme mannosidase. In the present case, diagnosis was made late in the illness after failure of a response to antipsychotic treatment and with the patient additionally showing progressive cognitive decline. Only after extensive investigation was the diagnosis made by showing decreased alpha-mannosidase enzyme activity in serum and blood leukocytes. This case demonstrates that an unusual clinical course or striking symptom patterns, especially in association with somatic comorbidity, in psychotic patients should lead to diagnostic consideration of inherited metabolic disease. PMID- 15759167 TI - [Diagnosis and differentiation of children with language development disorders. What role can be attributed to intelligence?]. AB - BACKGROUND: Specific language impairment (SLI) is defined as a developmental disorder in which language comprehension and the child's ability to use expressive spoken language is markedly below the appropriate level for his or her mental age. The intelligence of SLI children is in the normal range, while their language abilities are impaired. "Normal intelligence", the defining feature of SLI is questioned in this study. PATIENTS AND METHODS: Using IDIS (an inventory of diagnostic information in language impairment), we examined 138 children aged 5 and 6 years with severe language impairment; 108 SLI and 30 LI children. Various indicators of speech and language such as articulation, the ability to discriminate sounds, lexicon, grammar and pragmatic abilities but also auditory and visual perception, auditory and visual memory, fine and gross motor function were assessed. RESULTS: The performance of the SLI children was significantly higher in most of the tests than that of the LI children. Factor analysis showed that the two groups also differed in the structure of performance. Auditory short term memory was reduced in most children irrespective of intelligence. CONCLUSIONS: We propose the retention of the differentiation of subgroups of developmental speech and language disorders depending on the level of intelligence. PMID- 15759169 TI - Re-scaling social preference data: implications for modelling. AB - As applied in cost-utility analysis, generic health status indexes require that full health and dead are valued as 1 and 0, respectively. When social preference weights for health states are obtained using a visual analogue scale (VAS), their raw scores often lie on a scale with different endpoints (such as "best" and "worst" health). Re-scaling individual raw scores to a 0-1 scale leads to the exclusion of respondents who fail to value dead or full health. This study examined alternative approaches that do not impose such strict exclusion criteria. The impact of a different timing of re-scaling (before or after aggregation) and a different measure of central tendency (median or mean) is measured. Data from a postal valuation survey (n=722) conducted in Belgium are used. The following models are considered: (a) re-scaling values for EQ-5D health states on a within-respondent basis and using mean re-scaled values as proxies for social preference values, (b) using median re-scaled values as proxies for social preference values, (c) computing the median raw VAS values and then re scale, and (e) re-scaling mean raw VAS values. Exclusion rates, health state rankings and valuations and incremental value differences between pairs of states are computed for each model. Models that use a different timing of re-scaling, are compared ceteris paribus to evaluate the importance of timing of re-scaling and models that use a different measure of central tendency are compared ceteris paribus to evaluate the importance of the measure of central tendency. The exclusion rates are above 20% in the models that re-scale valuations before aggregation and less than 5% in the models that re-scale after aggregation. Health state valuations are found to be different in all two by two comparisons. Although in some comparisons the incremental values are statistically significantly different between models, they are never clinically significantly different. Differences in health state rankings were larger between the models that use a different measure of central tendency than between the models that re scale at a different time. This study shows that, for the data sample used, the choice of the measure of central tendency is more important for the social health status preference values than the timing of re-scaling. Using median original valuations in the analysis of EQ-5D valuation data is theoretically appealing because it builds on the "median voter" model and has the advantage of a lower respondent exclusion rate. Further analysis, on other data samples, will have to confirm the current findings. PMID- 15759170 TI - What reimbursement for coronary revascularization with drug-eluting stents? AB - We investigated the clinical and economic impact of sirolimus-eluting coronary stents (SES) at a nationwide level as to advice about the feasible reimbursement policy for the Italian Health Care System (SSN). A decision model compared bare metal stents (BMS) and SES in terms of costs and repeat coronary revascularizations incurred in 12 months following the first revascularization. The model was compiled for eight subgroups of patients. Rates of events were derived from randomized trials and an 1,809-patient survey. National charges were used to evaluate resources consumption. Compared with BMS, the number of averted revascularizations with SES is 0.16 per patient. SES also save Euro 1,371 per patient. Total savings to SSN are proportional to the rate of SES adoption by Italian hospitals: assuming a complete replacement of BMS with SES, the model estimates that 7,095 revascularizations would be averted and more than Euro 60 million saved by the SSN in 1 year. To stimulate SES adoption a SES-specific DRG might by introduced with a reimbursement value 23% higher than the current charge. SES is thus a cost-saving strategy in the perspective of the SSN that could therefore support the introduction of the new technology by reimbursing about 80% of its current incremental acquisition cost. PMID- 15759171 TI - Drug price regulation under consumer moral hazard. Two-part tariffs, uniform price or third-degree price discrimination? AB - Drug price differences across national markets as they exist in the EU are often justified by the concept of Ramsey prices: with fixed costs for R&D, the optimal mark-ups on marginal costs are inversely related to the price elasticity in the individual markets. This well-known result prevails if consumer moral hazard is taken into account. Contrary to the situation without moral hazard, the uniform price does not necessarily dominate discriminatory pricing in welfare terms. The two-part tariff is a better alternative as it allows governments to address moral hazard. A uniform price combined with lump-payments reflecting differences in the willingness to pay and the moral hazard in member states appears to be an attractive option for a common EU drug market. PMID- 15759168 TI - [Squamous cell carcinoma of the head and neck. Principles and current concepts of immunotherapy]. AB - Biologic therapies able to induce or up-regulate anti-tumor immune responses could represent a complementary approach to improve the conventional treatment of squamous cell carcinomas of the head and neck (SCCHN). Patients with SCCHN are frequently immunocompromised due to the elimination and dysfunction of critical immune effector cells. Therefore, it might be necessary to restore these immune functions to allow for the generation of effective anti-tumor host responses. Simultaneously, to prevent tumor escape from immunological recognition and destruction, it might also be necessary to alter antigenic and immunogenic attributes of the malignant cells. The present overview summarizes general aspects, historical data, and recent advances in the field of immunotherapy of SCCHN, including non-specific immune stimulation, transfer of immunocompetent cells, gene therapy, use of monoclonal antibodies, and anti-cancer vaccines. PMID- 15759172 TI - Back pain in pregnancy: 1-year follow-up of untreated cases. AB - Back Pain (BP) is one of the most frequent symptoms during the last period of pregnancy, and high incidence has been described in several studies. Until now no wide, multicenter and prospective clinical studies on the natural course of BP after pregnancy have been available. We performed a multicenter follow-up study in a sample of pregnant women using the Italian validated version of the Roland questionnaire to assess the evolution of BP after pregnancy and identify prognostic factors. Each center had to re-evaluate at least 75% of the initially enrolled women, with latency of 1 year after delivery. At the follow-up, we acquired substantial clinical data concerning the post-delivery period. The evaluation of symptom evolution was based on the Roland questionnaire. At follow up, 53% of re-evaluated women had no BP symptoms. Moreover, there was a significant improvement of patient-oriented assessment in women who suffered BP after delivery. With regard to the predictive factors, the presence of BP before pregnancy implied a 3.1-fold higher probability of improvement after delivery. In conclusion, women without history of BP before pregnancy and who complain of these symptoms during pregnancy require greater attention, because they have a lower possibility for improvement. Conversely, in women with a history of BP, pregnancy represents a transient period of worsening symptoms, probably due to the temporary para-physiological mechanical condition. PMID- 15759173 TI - Repositioning accuracy and movement parameters in low back pain subjects and healthy control subjects. AB - STUDY DESIGN: A control group study with repeated measures. OBJECTIVE: To compare trunk repositioning parameters in chronic low back pain (LBP) and healthy subjects. SUMMARY AND BACKGROUND DATA: Recent evidence suggests that chronic LBP patients exhibit deficits in trunk proprioception and motor control. Trunk repositioning and the various spatio-temporal parameters related to it can be used to evaluate sensori-motor control and movement strategies. METHODS: Fifteen control subjects and 16 chronic LBP subjects participated in this study. Subjects were required to reproduce different trunk position in flexion (15 degrees, 30 degrees and 60 degrees) and extension (15 degrees). In the learning phase preceding each condition, visual feedback was provided. Following these learning trials, subjects were asked to perform ten consecutive trials without any feedback. Movement time, movement time variability and peak velocity were obtained and a temporal symmetry ratio was calculated. Peak angular position variability and absolute error in peak angular position were also calculated to evaluate spatial accuracy. RESULTS: Two subgroups of LBP patients were identified. One subgroup of LBP subjects demonstrated longer movement time and smaller peak velocities and symmetry ratios than normal subjects. No group difference was observed for peak angular position variability and absolute error in peak angular position. CONCLUSION: Chronic LBP patients, when given a sufficient learning period, were able to reproduce trunk position with a spatial accuracy similar to control subjects. Some LBP subjects, however, showed modifications of movement time, peak velocity and acceleration parameters. We propose that the presence of persistent chronic pain could induce an alteration or an adaptation in the motor responses of chronic LBP subjects. PMID- 15759174 TI - Aplasia of the posterior arc of the atlas with persistent posterior tubercle: a case report. AB - An unusual case of partial aplasia of the posterior arc of the atlas, with persistent posterior tubercle, is presented in a previously healthy individual who sustained a neck trauma. Both plain X-rays and CT findings specified the lesion. Dynamic X-rays in flexion and extension showed an immobile posterior tubercle. The patient did not develop neurological symptoms at any stage during follow-up (1 year). PMID- 15759176 TI - Gastrojejunal strictures following laparoscopic Roux-en-Y gastric bypass for morbid obesity. AB - BACKGROUND: Gastrojejunal strictures following laparoscopic Roux-en-Y gastric bypass (LRYGBP) present with dysphagia, nausea, and vomiting. Diagnosis is made by endoscopy and/or radiographic studies. Therapeutic options include endoscopic dilation and surgical revision. METHODS: Of 369 LRYGBP performed, 19 patients developed anastomotic stricture (5.1%). One additional patient was referred from another facility. Pneumatic balloons were used for initial dilation in all patients. Savary-Gilliard bougies were used for some of the subsequent dilations. RESULTS: Flexible endoscopy was diagnostic in all 20 patients allowing dilation in 18 (90%). Two patients did not undergo endoscopic dilation because of anastomotic obstruction and ulcer. The median time to stricture development was 32 days (range: 17-85). Most patients (78%) required more than two dilations. The complication rate was 1.6% (one case of microperforation). At a mean follow-up of 21 months, all patients were symptom-free. CONCLUSIONS: Gastrojejunostomy stricture following LRYGBP is associated with substantial morbidity and patient dissatisfaction. Based on our experience, we propose a clinical grading system and present our strategy for managing gastrojejunal strictures. PMID- 15759177 TI - A new device for bone anchor fixation in laparoscopic sacrocolpopexy: the Franciscan laparoscopic bone anchor inserter. AB - BACKGROUND: A laparoscopic modification of the sacrocolpopexy procedure with mesh and bone anchor fixation with the Franciscan laparoscopic bone anchor inserter was developed. METHODS: We developed a laparoscopic bone anchor inserter for the placement of a titanium bone anchor in sacral segment 3 as fixation for the mesh in laparoscopic sacrocolpopexy procedures performed in women with posthysterectomy vault prolapse. RESULTS: Surgery successfully corrected vaginal vault prolapse. Laparoscopic bone anchor insertion with this new and simple device took 2 minutes and provided a firm anchor for mesh fixation. MRI demonstrated an anatomically preferable vaginal axis toward the hollow of the sacrum. CONCLUSION: Application of the newly developed Franciscan laparoscopic bone anchor inserter in laparoscopic sacrocolpopexy is an easy and safe procedure that provides firm fixation and excellent anatomical results. PMID- 15759178 TI - Early results of a randomized multicenter trial comparing Prolene and VyproII mesh in bilateral endoscopic extraperitoneal hernioplasty (TEP). AB - BACKGROUND: The use of mesh in inguinal hernia surgery has become increasingly popular despite the potentially harmful effects that foreign material may exert on human tissue. The purpose of this study was to compare the use of a lightweight mesh vs a standard hernia mesh in bilateral endoscopic hernioplasty. METHODS: This single-blinded multicenter trial, 140 men with bilateral inguinal hernias were randomized to undergo totally extraperitoneal endoscopic hernioplasty (TEP) with either Prolene or VyproII. The randomization and all data management were done via the Internet. RESULTS: A total of 139 patients were operated on as allocated. The follow-up was complete in 94% of the patients. The median (range) operating times were 85 min (45-140) and 73 min (35-165) for the Prolene and VyproII groups, respectively. (p = 0.01). The difference was due to uneven distribution of the allocated patients to study groups among individual surgeons. The time to return to work was similar (11 vs 9 days, p = 0.08). The time to return to normal daily activities was 19 days (1-133) in the Prolene group and 12.5 days (0-237) in the VyproII group (p = 0.06). There were no significant differences between the groups in their scores on the Visual Analogue Scale or SF-36 Health Scores during the 8-week follow-up. CONCLUSIONS: The use of Prolene and VyproII meshes in bilateral endoscopic repair of inguinal hernia seems to result in similar short-term outcome and quality of life. However, there was a tendency toward faster return to normal activity among VyproII patients. PMID- 15759179 TI - Improving the safety of laparoscopic cholecystectomy: the routine use of preoperative magnetic resonance cholangiography. AB - BACKGROUND: The aim of this study was to determine the value of routinely performed preoperative magnetic resonance cholangiography (MRC) in detecting common bile duct (CBD) stones in patients stated to undergo elective laparoscopic cholecystectomy. In addition, we used MRC to investigate possible variants of the cystic duct. METHODS: Magnetic resonance cholangiography was performed preoperatively in 773 patients (311 male and 462 female; median age 55 years, range 16-91) who had no clinical signs of cholestasis prior to undergoing elective laparoscopic cholecystectomy. In cases where the MRC was positive for CBD stones, endoscopic retrograde cholangiopancreatiography (ERCP) was then performed. A total of 532 patients were available for continuous postoperatively follow-up (median 54 months, range 36-85). In 462 patients (247 female, and 215 male), MR images were also reviewed for variants of the cystic duct. RESULTS: In 705 patients (91%), MRC was negative for CBD stones. In 64 patients (9%) MRC was positive. Of these patients, 47 (6%) had CBD stones on ERCP. In 12 patients (2%), MRC was false positive. In five cases (0.6%), ERCP had an inconclusive result postoperative follow-up (532 patients, or 69%) revealed evidence of CBD stones in three patients (10.4%) despite a preoperative negative MRC result. Anatomical variants in the course of the cystic duct and its confluence with the common bile duct were found in 27 of 462 patients (6%). CONCLUSIONS: Magnetic resonance cholangiography proved to be a reliable screening technique in the preoperative evaluation of patients with silent CBD stones. Imaging of the course of the cystic duct is possible in a high percentage of cases. Therefore, MRC can be recommended as a screening technique before laparoscopic cholecystectomy. PMID- 15759181 TI - Endoscopic adrenalectomy: an analysis of the transperitoneal and retroperitoneal approaches and results of a prospective follow-up study. AB - BACKGROUND: Endoscopic adrenalectomy is currently performed using either a retroperitoneal or transperitoneal approach. The aim of this study was to determine which of these is the optimal surgical technique in a prospectively designed analysis of a large series of patients operated on by a single team over a 10-year period. METHODS: From February 1994 to March 2004, 267 endoscopic adrenalectomies (retroperitoneal in 132 patients and transperitoneal in 135 patients) were performed in 245 consecutive patients. There were 102 right lateral and 121 left lateral procedures (22 patients had a bilateral procedure). The most prevalent indication was incidentaloma (35.9%), followed by pheochromocytoma and Conn's adenoma. RESULTS: The endoscopic procedure was performed in 238 of 245 patients (97.1%). The conversion rate was 1.5% for the transperitoneal approach and 3.8% for the retroperitoneal approach. No statistically significant influence was noted for the parameters of intraoperative blood loss, rate of postoperative complications, and duration of hospital stay with regard to the surgical technique. The operative time and the learning curve proved to be significantly longer for the retroperitoneal adrenalectomy. In addition, a variance analysis identified tumor size (>5 cm) as a significant factor influencing the operative time, whereas body mass index and localization (right/left lateral) did not prove significant. CONCLUSION: Independent of the underlying pathology, endoscopic adrenalectomy using either the trans- or retroperitoneal approach can be performed in 96-98% of all patients. Differences between the two techniques in operative time and learning curves clearly favor the transperitoneal adrenalectomy. PMID- 15759180 TI - Laparoscopic rectopexy for full-thickness rectal prolapse: a single-institution retrospective study evaluating surgical outcome. AB - BACKGROUND: The laparoscopic approach promises to become the gold standard for the transabdominal management of full-thickness rectal prolapse. The aim of this study was to review our experience and to highlight the functional results achieved with this new technique. METHODS: Forty-eight patients with full thickness external prolapse underwent laparoscopic repair between February 1997 and February 2003. All patients underwent preoperative evaluation of their rectal function. Patients with isolated rectal ulcer without prolapse or with internal prolapse and patients deemed by the anesthesiologist to be unfit for general anesthesia were excluded from the study. The laparoscopic technique was either a mesh rectopexy without resection (n = 35) or a suture rectopexy with sigmoid resection (n = 13). Patients with intractable constipation preceding the development of the rectal prolapse were advised to have a resection-rectopexy. In the postoperative follow-up, attention was paid to mortality, morbidity, recurrent prolapse, incontinence, and constipation. Follow-up was done by clinical review and postal questionnaire. RESULTS: There were no deaths and no septic or anastomotic complications. The postoperative morbidity rate was 5%. Oral intake was started on postoperative day 1. Discharge from the hospital was on postoperative day 4 in patients without sigmoid resection and on postoperative day 7 in patients with sigmoid resection. Two patients (4%) developed recurrent total prolapse during a median follow-up period of 36 +/- 15 months (range, 7 77). The functional results were good or excellent in 72% of the cases, without digitations or dyschesia. Continence was improved in 31% of the patients and remains unchanged in 64% of them. In 11 patients (23%), constipation was worsened by the procedure. CONCLUSION: Laparoscopic rectopexy with or without resection is both safe and effective. Advantages include low-morbidity, improved cosmesis, the rapid return of intestinal function, early discharge from hospital, and a low recurrence rate. The fecal continence score is improved; however, constipation is frequently worsened. PMID- 15759182 TI - Radiofrequency ablation in the liver close to the bile ducts: can intraductal cooling offer protection? AB - BACKGROUND: One complication of radiofrequency ablation (RFA) of the liver is biliary duct damage. Intraductal cooling (IDC) has been proposed as a means of protection. METHODS: In the first experiment, designed to evaluate the influence of IDC on the RFA procedure per se and on lesion formation, lesions were created in vivo in pig liver with and without IDC. The RFA needle was placed with a 1.5 cm safety margin from the bile ducts. In the second experiment, designed to evaluate the potential protective effects of IDC, lesions were created close to a bile duct with and without IDC. RESULTS: With the safety margin, the RFA parameters and lesion size were not negatively affected by IDC. Microscopic examination revealed that IDC had a protective effect in most of the lesions created close to a bile duct. CONCLUSIONS: The IDC procedure was feasible and had no negative effect on the RFA procedure or the lesions. However, the protective effect of IDC was not statistically significant (p = 0.12). PMID- 15759183 TI - The economic impact of incorporating flexible endoscopy into a community general surgery practice. AB - BACKGROUND: Flexible endoscopy is a vital component of gastrointestinal surgery. It has and will replace many of the surgical procedures now commonly performed. Flexible endoscopy, unfortunately, is not an integral part of surgical residency training based on resident operative experience as reported by the Residency Review Committee. Moreover, general surgeons have deferred the practice of flexible endoscopy to the gastroenterologists because of concerns over turf battles and referral patterns. The purpose of this study was to assess the overall case load and the economic impact of flexible endoscopy on the practice of general surgery in a community hospital setting. METHODS: This retrospective review was performed over a 6-month period. The total cases and the total billings of inpatient and outpatient procedures for a group practice of five general surgeons in a community hospital were evaluated. The billings were the actual charges based on current procedural terminology (CPT) codes for these procedures using the Medicare fee schedule. RESULTS: Of the 2,159 procedures performed, 1,154 involved flexible endoscopy cases accounting for 54% of all cases (1,154 of 2,159) performed from February 1, 2003 to July 31, 2003. Flexible endocopy accounted for 43% of the total charges. A. total of 46 surgical procedures and 216 future endoscopies were generated from the flexible endoscopic procedures. Future endoscopic cases were for surveillance of colonic neoplasia and Barrett's esophagus. CONCLUSIONS: Flexible endoscopy contributed to a major portion of the caseload and revenue generated by the general surgery group studied. The overall impact of flexible endoscopy is even greater than reported because of the future endoscopic surveillance cases or surgical interventions generated on the basis of endoscopic findings. PMID- 15759184 TI - Comparison of the benefits and complications between laparoscopic and open Roux en-Y gastric bypass surgeries. AB - BACKGROUND: In recent years, there has been an increase in numbers of individuals seeking laparoscopic surgical procedures for obesity. The current study compared the benefits and risks between laparoscopic and open Roux-en-Y gastric bypass (RYGBP) performed at the same center for more than 2,000 patients. METHODS: The study population consisted of 1,077 laparoscopic and 1,198 open RYGBP procedures performed between the years 1999 and 2002. Measurements included population characteristics, anthropometries, complications, and hospital stay. RESULTS: The laparoscopic RYGBP has both advantages and disadvantages. The disadvantages include a longer operative time and a higher incidence of fistulas, internal hernias, and small bowel obstruction. The advantages of the laparoscopic procedure are shorter hospital stay, lower incidence of wound infection, and fewer incisional hernias. Both procedures cause similar changes in body weight, but laparoscopic RYGBP is associated with less lean tissue loss during the early postoperative period. CONCLUSION: Both laparoscopic and open RYGBP are effective in inducing massive weight loss. There are, however, differences in the benefits and risks between the two procedures. PMID- 15759185 TI - National trends in utilization and outcomes of bariatric surgery. AB - BACKGROUND: Because of the growing interest in surgery to treat morbid obesity, this study examined changes in the utilization and in-hospital outcomes of bariatric surgery in the United States over a 10-year period. METHODS: Data were obtained from the Nationwide Inpatient Sample, the largest all-payer discharge database in the United States. International Classification of Disease (ICD-9) codes were used to identify all bariatric procedures performed for adults from 1990 to 2000. Population-based rates of surgery for each year were calculated by applying sampling weights and U.S. Census data. Secular trends in annual rates of surgery, changes in patient characteristics, and in-hospital mortality and complications were analyzed. RESULTS: From 1990 to 2000, the national annual rate of bariatric surgery increased nearly six fold, from 2.4 to 14.1 per 100,000 adults (p = 0.001). There has been more than a ninefold increase in the use of gastric bypass procedures (1.4 to 13.1 per 100,000; p < 0.001). This represents an increase from 55% of all bariatric procedures in 1990 to 93% of such procedures in 2000 (p < 0.001). The rates of in-hospital mortality were low (0.4% overall), but increased slightly over time (0.2% in 1990 to 0.5% in 2000; p = 0.009). There is no significant difference in adjusted mortality for the past 8 years, but a slight rise did occur over the full 10-year period. The rates for reoperation (1.3%) and pulmonary emboli (0.3%) remained stable. The rates for respiratory failure associated with bariatric surgery declined from 7.7% in 1990 to 4.5% in 2000 (p < 0.001). Over this time, the mean length of hospital stay declined from 6.0 to 4.1 days (p < 0.001). CONCLUSIONS: The annual rate of bariatric surgery in the United States increased nearly six fold between 1990 and 2000, with little change in in-hospital morbidity and mortality. This appears to be driven largely by the increasing popularity of gastric bypass procedures. PMID- 15759186 TI - Predictors of survival after laparoscopic radiofrequency thermal ablation of hepatocellular cancer: a prospective study. AB - BACKGROUND: Most patients with hepatocellular carcinoma (HCC) are not candidates for hepatic resection or liver transplantation. Radiofrequency ablation (RFA) provides local control for unresectable HCC with minimal morbidity. The aim of this prospective study is to determine factors predicting survival in patients with HCC undergoing RFA. METHODS: Sixty-six consecutive patients with HCC who were not candidates for a curative liver resection and were free of extrahepatic disease underwent laparoscopic RFA. The relationship between demographic, clinical, laboratory, and surgical parameters and survival was assessed using univariate Kaplan-Meier survival and multivariate Cox proportional hazards model. RESULTS: The median Kaplan-Meier survival for all patients was 25.3 months after RFA. Although alfa fetal protein (AFP), bilirubin, ascites, and Child class were statistically significant predictors of survival by univariate analysis, only the Child class and AFP were independent predictors by multivariate analysis. CONCLUSIONS: This study determines which patients do best after RFA and shows that RFA can provide significant survival for patients with unresectable HCC while also forming a bridge to liver transplantation. RFA has become the first line of treatment in the management of these patients. PMID- 15759187 TI - Tisseel versus tack staples as mesh fixation in totally extraperitoneal laparoscopic repair of groin hernias: a retrospective analysis. AB - BACKGROUND: The laparoscopic repair of groin hernias generally involves mesh fixation to avoid displacement and recurrence. Fixation usually uses staples that can lead to nerve injury and chronic postoperative pain. Laparoscopic repairs are associated with a risk of chronic pain of up to 22.5%. The use of fibrin glue (Tisseel) may represent an alternative method of mesh fixation preventing the risk of nerve injury. METHODS: Sixty-six patients had groin hernia repair using a totally extraperitoneal (TEP) laparoscopic procedure. Mesh fixation was achieved using 2 ml of fibrin glue. Comparison was made with an earlier series of 102 patients operated on according to the same procedure in which mesh fixation used tack staples. Complications, length of stay, recurrence, and postoperative chronic pain were assessed. RESULTS: No difference was found between the two series, except there was a significantly higher rate of postoperative chronic pain in the staples series (14.7 vs 4.5%, p = 0.037) and there was one recurrence (1.5%) in the fibrin glue group of patients. CONCLUSIONS: Fibrin glue achieved an adequate mesh fixation with a lower incidence of chronic postoperative pain. Although a prospective randomized study is needed, Tisseel appears to be an alternative to staples for mesh fixation and may help reduce the postoperative pain problems after hernia repair. PMID- 15759188 TI - Laparoscopic vs open total colectomy: a case-matched comparative study. AB - BACKGROUND: Open total colectomy and ileorectal anastomosis (OTC) is a major colorectal procedure which would preclude laparoscopy in many centers because of technical difficulty and the fact that laparoscopic total colectomy (LTC) takes much longer than standard laparoscopic proctosigmoidectomy (LPS). This study compares OTC with LTC and LPS. METHODS: In this study, 34 LTC patients (May 1999 to August 2003) were matched for age, diagnosis, operative period, and procedure with patients undergoing OTC. Patients with a previous major laparotomy were excluded from the open group. Groups were compared for gender, American Society of Anesthesiology (ASA) classification, operating time, estimated blood loss, length of hospital stay (LOS), complications including readmissions, and costs. The LPS cases were picked randomly from the laparoscopic database (every eighth patient), and the OT and LOS were noted. RESULTS: The LTC and OTC groups were matched for age (mean, 31 vs 34 years; p = 0.2), sex (14 vs 13 females; p = 0.8), ASA (8/23/3/0 vs 8/22/4/0, class 1/2/3/4). The body mass index was higher in the open group (23.8 vs 27.9; p = 0.04). The operating time was significantly longer (187 vs 126 min; p = 0.0001) and the median LOS shorter in the LTC group (3 days [IQR, 2.5-5 days] vs 6 days [IQR 4-8 days]; p = 0.0001). The estimated blood loss was significantly less in the LTC group (168 [50-700] ml) vs 238 [50-800] ml); p = 0.001, but there was no significant difference in the complication (26.5% vs 38.2%; p = 0.4) readmission (11.8% vs 14.7%; p = 1.0), reoperative rates (8.8% vs 11.8%; p = 1.0), or direct costs ($4,578 vs $4,562; p = 0.3). One LTC patient died expired on postoperative day 2 of a cardiac event. Four patients (11.8%) required conversion for obesity (n = 2), adhesions (n = 1), or intraoperative hemorrhage (n = 1). The operating times were 36 min longer in the LTC group than in the LPS group (151 vs 187 min; p = 0.02), but there was no significant difference in the LOS. (3 vs 3 days, p = 0.2). CONCLUSIONS: The findings show that LTC provides a significant decrease in the LOS over OTC, with increased operating time, but without any change in other parameters. A laparoscopic approach to subtotal colectomy is recommended for suitable patients when an experienced team is available. PMID- 15759191 TI - Seeing is believing: visualization systems in endoscopic surgery (video, HDTV, stereoscopy, and beyond). PMID- 15759189 TI - Does routine intraoperative cholangiography prevent bile duct transection? AB - BACKGROUND: The role of routine intraoperative cholangiography is controversial. The aim of this study was to assess the impact of routine intraoperative cholangiography on the incidence of common bile duct injuries, and to evaluate the operative outcome of laparoscopic cholecystectomy carried out in a major teaching hospital and review the literature. METHODS: Prospectively collected data on 3,145 laparoscopic cholecystectomies performed mainly by surgical trainees in the period 1990 to 2002 using routine intraoperative cholangiography with fluoroscopy were reviewed. RESULTS: The mean age of the study sample (65.6% male, 34.4% female) was 54 years, and 16.9% of the patients had clinical acute cholecystitis. The conversion rate to open cholecystectomy was 4.3%. Intraoperative cholangiography was attempted for 90.7% of the patients with a 95.9% success rate. Five patients (0.16%) had common bile duct injuries. Four injuries had occurred in the first 5 years. One injury (0.06%) had occurred after 1995. This injury was identified intraoperatively and repaired laparoscopically. Routine intraoperative cholangiography prevented one definite common bile duct transection. CONCLUSIONS: In this series using routine intraoperative cholangiography, there was a low rate and severity of common bile duct injuries, with a high intraoperative recognition rate. There was no bile duct transection or major injury requiring common bile duct reconstruction. Although intraoperative cholangiography helped in the immediate identification of injuries and the institution of appropriate therapy, injury was not completely prevented. PMID- 15759190 TI - Laparoscopic colorectal resection for endometriosis. AB - BACKGROUND: The rectosigmoid colon is affected by deep pelvic endometriosis in 3 37% of cases. In the past, treatment of the affected gastrointestinal tract generally required conversion to conventional surgery. We describe our experience with complete laparoscopic management of deep pelvic endometriosis with bowel involvement. METHODS: From March 1995 to March 2003, 29 consecutive patients with endometriosis requiring laparoscopic intervention were evaluated. In seven patients (24%) colorectal involvement was identified prior to the operation. A low anterior resection was performed in four patients (57%) and a sigmoid resection in three (43%). In all cases, colonoscopy showed a normal mucosa. In all cases, treatment consisted of resection of the bowel involved together with the excision of all other implants. Data analysis included age, previous abdominal operations, previous history of endometriosis, operative time, conversion rate, complications, length of stay, and pain relief. RESULTS: There were seven patients with colorectal involvement whose median age was 32.8 years (range, 28-40), with a history of previous abdominal operation in two (28%). Preoperative symptoms were as follow: dysmenorrea in four patients (57%), dyspareunia in four (57%), pelvic pain in seven (100%), rectal bleeding in one (14%), and tenesmus in five (71%). Mean operative time was 190 min (range, 165 230). Length of stay was 8.3 days (range, 7-11). There were no anastomotic leak and no major postoperative complication. One patient had temporary urinary retention. At a median follow-up of 38.7 months (range, 1-84), complete relief of pelvic symptoms was achieved in five patients (71%), and there was improvement in one patient. In one patient complaining of persistent pain, a new colonic implant was diagnosed two years after the surgery requiring reoperation. CONCLUSIONS: The results show that provided that the surgeon is highly skilled in laparoscopy, laparoscopic resection of deep pelvic endometriosis with rectosigmoid involvement is feasible and effective in nearly all patients. PMID- 15759192 TI - Headmount stereoscopic monitor for endoscopic surgery. PMID- 15759193 TI - A simple technique for circular-stapled Billroth I reconstruction in laparoscopic gastrectomy. AB - A simple surgical technique that facilitates circular-stapled Billroth I gastroduodenostomy in laparoscopic distal gastrectomy is described. After standard laparoscopic mobilization of the distal stomach, a small duodenotomy is made just distal to the pyloric ring. The anvil of a circular-stapling device, secured with a Vicryl suture, is introduced via the duodenotomy. The Vicryl suture is advanced anteriorly so that a center rod penetrates the anterior duodenal wall. The duodenum is staple-transected at this point, and the center rod is wrapped with the stapled duodenal stump by approximation of both edges using a suturing device. The circular-stapled gastroduodenostomy then is completed in a standard fashion. The authors have used this technique for three patients, and their early outcomes are promising. PMID- 15759194 TI - Long-term behavioral assessment of guinea pigs following neonatal pneumoperitoneum. AB - BACKGROUND: Using guinea pigs, we previously demonstrated that pneumoperitoneum during pregnancy produces behavioral deficits in the offspring. In the current study, the purpose was to determine if CO(2) pneumoperitoneum during the early postnatal period also produced behavioral anomalies. METHODS: Following delivery, guinea pig pups were randomly assigned to one of three treatment groups: CO(2) pneumoperitoneum (P), laparotomy (L), or isolation control (I). Surgeries were performed on postnatal day (PND) 5 under isoflurane anesthesia; control pups were isolated from the dams for an equivalent period of time. On PNDs 10, 20, 40, and 60, behavior was assessed by monitoring locomotor and exploratory activity. RESULTS: A total of 29 animals were studied. We observed no immediate morbidity or mortality and the manipulations did not appear to affect postnatal growth. On PND 10, pups in group P exhibited lower levels of locomotor activity compared to L and I neonates, but this difference resolved as the animals got older. Histologic assessment of the adult offspring brains revealed no evidence of neurologic injury. CONCLUSION: These data suggest that unlike insufflation during pregnancy, neonatal pneumoperitoneum does not produce behavioral deficits. PMID- 15759196 TI - Self-reported versus observed scores in laparoscopic skills training. AB - BACKGROUND: Education in basic laparoscopic skills training is performed ideally in an inanimate laboratory. Monitoring of basic skills progression, which is essential during this tranining, often may be difficult because of the resources and manpower. This study investigated the differences between self-reported and observed scores during basic laparoscopic exercises. METHODS: First-and second year medical students involved in practice sessions using the LTS 2000 box trainer were included in the study. All the students were allowed to practice four tasks: placing pegs with their dominant and nondominant hands, transferring pegs from one hand to another starting with the dominant and nondominant hands, cannulating a pipe cleaner into a plastic tube, and progressing from one end of a rope to the other. Each student recorded his or her times and errors for each task. After these practice sessions, each student underwent an observed session (OS), performing all of the tasks, and was graded by a surgeon involved in laparoscopic education. All the students were asked to record another practice session. The self-reported performances from the session immediately before the OS (pre-OS) and the session immediately after the OS (post-OS) were compared with the performance in the OS. RESULTS: A total of 36 students were included in this study. The mean time and mean error were similar between all the sessions. Although the scores in the OS did slightly differ from the scores in the pre-OS and post-OS, post hoc analysis showed that there was no statistically significant difference between the OS score and either the pre-OS or the post-OS score for any of the laparoscopic exercises. CONCLUSIONS: No differences between observed scores and self-reported scores were noted. The use of self-reported scores may be a good method for monitoring performance during training in laparoscopic basic skills. The self-reporting of basic laparoscopic exercises may conserve resources. PMID- 15759195 TI - The impact of laparoscopy on bariatric surgery. AB - The rising popularity of bariatric surgery over the past several years is attributable in part to the development of laparoscopic bariatric surgery. Morbidly obese patients have associated comorbid conditions that may predispose them to postoperative morbidity. The laparoscopic approach to bariatric surgery offers a minimally invasive option that reduces the physiologic stress and provides clinical benefits, as compared with the open approach. This review summarizes the impact of laparoscopic surgery on bariatric surgery, the various risk factors that could potentially predispose morbidly obese patients to postoperative morbidity, the fundamental differences between laparoscopic and open bariatric surgery, and the physiology of reduced tissue injury associated with laparoscopic bariatric surgery. PMID- 15759198 TI - Experimental assessment of a new mechanical endoscopic solosurgery system: Endofreeze. AB - BACKGROUND: The assistance received by the surgeon from support personnel during operative laparoscopy is extremely important. This includes retraction of instruments and endoscope positioning. However, human assistance is costly and often does not provide satisfaction for the surgeon. The aim of this study was to develop a mechanical arm capable of allowing easy handling and holding of laparoscopic instruments under the surgeon's control. METHODS: We designed a system, named Endofreeze, based on a particular kinematical construction that maintains an invariant point of constraint motion just above the trocar puncture site through the abdominal wall. The goal was to develop this way a highly intuitive mechanical holding system for laparoscopic instruments, with sufficient precision of action, activated by a single hand movement. We tested a couple of prototypes with different holding arms while performing cholecystectomy in phantom models with swine inserts and compared the results obtained in similar conditions using different holding and positioning systems. RESULTS: The system allows transparent and intuitive operation, and its setup is easy and quick. It may be adapted either as an instrument retractor or as an optic positioning device. Compared to different systems available or prototypes previously tested, such as AESOP 2000, ENDOASSIST, FIPS Endoarm, TISKA Endoarm, and the Martin Arm, in similar conditions, it was more intuitive, allowing shorter time for completion of surgery. CONCLUSION: Endofreeze is a new intuitive mechanical positioning system for endoscopic solo surgery. In phantom models, it demonstrated a shorter time requirement for completion of surgery when compared to other systems available. In our opinion, it represents a valid compromise between human and robotic control for conventional laparoscopic instruments. PMID- 15759197 TI - The value of endoscopic diagnosis and the treatment of pancreas injuries following blunt abdominal trauma. AB - BACKGROUND: Injuries to the pancreas following blunt abdominal trauma are rare due to its protected retroperitoneal position. Many pancreatic lesions remain unnoticed at first and only become apparent when complications arise or during treatment of other injuries. The mortality rate is between 12 and 30%, and if treatment is delayed it is as high as 60%. METHODS: Using medical records over the past 5 years, we investigated when and in what circumstances endoscopic retrograde cholangiopancreaticography (ERCP) was used in the diagnosis and treatment of pancreas injuries after blunt abdominal trauma. Penetrating injuries were not taken into consideration. RESULTS: An ERCP was performed on a total of five patients with suspected injuries to the pancreas after blunt abdominal trauma. No duct participation could be determined in three of the patients with a first degree pancreatic lesion. A 44-year-old woman sustained severe internal and external injuries after a traffic accident. Because of the nature of her injuries, pancreatic left resection with splenectomy was necessary. After the operation, a pancreatic fistula diagnosed. The ductus pancreaticus (DP) was successfully treated by stenting with the use of endoscopic retrograde pancreaticography. A 24-year old woman was kicked in the epigastrium by a horse. On the day after the incident, she complained of increasing pain in the upper abdomen, and she had elevated amylase and lipase levels. Computed tomography scan showed free fluid. Less than 48 h after the accident, ERCP was performed and a leakage in the DP in the head-body region (fourth degree) was identified. We placed a stent, and during the subsequent laparoscopy the omental bursa was flushed out and a drainage laid. After 14 days, the patient was sent home. We removed the drainage 4 weeks after the accident, and the stent after 12 weeks. CONCLUSION: The major advantage of the prompt retrograde discription of the pancreatobiliary system after an accident in which pancreas involvement is suspected is the more precise assessment of the extent of the injuries. If a stent is placed in the same session, it is possible to carry out definitive and interventional treatment. PMID- 15759199 TI - Hand dominance and performance in a laparoscopic skills curriculum. AB - BACKGROUND: This study evaluated the influence of hand dominance on skill acquisition during a basic laparoscopic skills curriculum. METHODS: A total of 27 surgical residents (5 postgraduate year 3 [PGY-3] and 22 PGY-2 residents) participated in a 4-week laparoscopic skills curriculum. The residents were pre- and posttested on six laparoscopic tasks during weeks 1 and 4. During weeks 2 and 3, the residents attended a proctored practice session. The results were compared using analysis of variance (ANOVA), (with significance determined by a p value less than 0.05. RESULTS: The posttest scores were significantly higher than the pretest scores. On the pretest, lefthand-dominant (LHD) surgeons (n = 4) performed significantly better than righthand-dominant (RHD) surgeons (n = 23). In the analysis of individual task pretest scores, LHD surgeons performed significantly better on pattern cutting and vessel loop application. Posttest analysis of overall performance did not show significant differences between the RHD and LHD surgeons. CONCLUSIONS: Participation in a laparoscopic skills curriculum improved overall performance. The LHD surgeons demonstrated better initial performance, but posttest comparison showed no difference between the two groups. PMID- 15759200 TI - Unusual imaging appearances of pancreatic serous cystadenoma: correlation with surgery and pathologic analysis. AB - BACKGROUND: We describe imaging and pathologic features of serous cystadenoma of the pancreas on multislice helical computed tomography CT (MS-CT) and surgical resection. METHODS: Radiologic and pathologic features were analyzed in five patients. All patients underwent MS-CT and digital subtraction angiography (DSA), and four patients underwent magnetic resonance (MR) imaging. Preoperatively, three cases showed radiologic evidence of mainly solid appearance on MS-CT, and the suspected diagnoses were solid pancreatic tumors (patients 1-3). The other two cases showed radiologic evidence of macrocystic tumor of the pancreas, and the suspected diagnoses were mucinous cystic tumors (cases 4 and 5). All patients underwent surgery, and the diagnosis of serous cystadenoma was confirmed on pathologic examination. RESULTS: In three cases that showed a solid appearance on MS-CT, a microcystic appearance was identified on microscopic examination, and the tumors were found to be hypervascular lesions on multiphasic contrast enhanced CT and DSA. In cases 1 and 2, the lesions showed high intensity with internal septation on T2-weighted MR images. In two cases, the tumors were classified as a macrocystic variant of serous cystadenoma, and no mural nodules, papillary projections, or calcifications were seen in the tumors. CONCLUSION: Imaging appearance of serous cystadenoma on MS-CT is various and sometimes indistinguishable from that of solid tumor or mucinous cystic tumors of the pancreas. Imaging findings of hypervascularity and a well-marginated high intensity lesion with internal septation on T2-weighted MR imaging may be crucial to identify serous cystadenoma that contains no visible cystic compartments on MS CT. PMID- 15759201 TI - Pseudoephedrine and guaifenesin urolithiasis: widening the differential diagnosis of radiolucent calculi on abdominal radiograph. AB - Unenhanced helical computed tomography has played an increasingly important role in the management of urinary tract stones, guiding diagnosis and control of calculus disease. We report computed tomographic and radiographic appearances of a renal calculus composed of pseudoephedrine and guaifenesin in a patient who abused over-the-counter allergy medication. PMID- 15759203 TI - Dark lumen MR colonography based on fecal tagging for detection of colorectal masses: accuracy and patient acceptance. AB - BACKGROUND: Magnetic resonance colonography (MRC) with fecal tagging has recently been investigated in clinical studies for the detection of polyps. We assessed fecal tagging MRC in a field trial. METHODS: Forty-two patients in a private gastroenterologic practice underwent MRC with barium-based fecal tagging (150 mL of 100% barium at each of 6 main meals before MRC) and conventional colonoscopy. Diagnostic accuracy of MRC and patient acceptance were assessed and compared with the respective results of conventional colonoscopy. RESULTS: Eighteen percent of all MRC examinations showed a remaining high stool signal in the colon that impeded a reliable inclusion or exclusion of polyps. On a lesion-by-lesion basis, sensitivities for polyp detection were 100% for polyps larger than 2 cm (n = 1), 40% for polyps between 10 and 19 mm, 16.7% for polyps between 6 and 9 mm, and 9.1% for polyps smaller than 6 mm. The main reason for the low acceptance of MRC was the barium preparation, which was rated worse than the bowel cleaning procedure with conventional colonoscopy. CONCLUSION: MRC with fecal tagging must be further optimized. The large amount of barium resulted in poor patient acceptance, and barium according to this protocol did not provide sufficient stool darkening. Other strategies, such as increasing the hydration of stool, must be developed. PMID- 15759204 TI - Radiologic imaging of the transplanted bowel. AB - BACKGROUND: The radiologic evaluation of the transplanted bowel is largely unknown and rather complex because it involves several techniques that depend on indications and times that have not been fully defined. METHODS: From December 2000 to November 2002 in the Section of Radiology I of the University of Modena and Reggio Emilia (Modena, Italy), 11 patients with transplanted bowel were studied with different methods: traditional radiologic evaluation with contrast agent (all patients), evaluation of transit time with radiopaque markers (five patients), ultrasonographic (US) evaluation of the intestinal wall and Doppler US of the vascular axes (five patients), computed tomographic (CT) evaluation (all patients), and magnetic resonance (MR) evaluation of the bowel and the vascular axes (five patients). Traditional contrast examination enabled evaluation of the gastroesophageal transit and cardia functionality; anatomy and integrity of the anastomoses (proximal and distal); time of gastric emptying; morphology, tone, and kinesis of the transplanted small bowel loops and time of global transit. The study of transit with radiopaque markers was carried out in five patients to define the time of transit through the entire transplanted bowel, confirm recovery of intestinal motility, and identify possible abnormalities. The US examination was carried out in five patients to evaluate the morphology, thickness, and echo structural features of the intestinal loops. Color Doppler was performed to visualize the superior mesenteric artery and a wall arteriole of the sampled loop. CT examination was performed 2 to 4 weeks after surgery to evaluate the anatomy of the transplanted organs, arterial and venous anastomoses in case of complications identified with other methods or suspected, and periodically in the follow-up of patients who underwent transplantation due to Gardner syndrome. The protocol for MR evaluation of the bowel included coronal single-shot fast spin-echo T2-weighted sequences, axial and/or sagittal single shot fast spin-echo T2-weighted sequences, coronal fast multiplanar spoiled gradient-echo (FMP- SPGR) sequences, coronal FMPSPGR sequences with and without administration of intravenous paramagnetic contrast agent, and axial or sagittal FMPSPGR fat-saturated sequences performed after dynamic gadolinium administration. RESULTS AND CONCLUSION: The study of transit with radiopaque markers was useful in patients with chronic intestinal pseudo-obstruction because it identified recovery and normalization of motility. Traditional contrast examination of the gastrointestinal tract continues to play an important role in transplanted patients because it is a simple examination that allows evaluation of the graft anatomy and recovery of motility of the residual native bowel and the transplanted loops. Moreover, it plays a crucial role in early detection of major postoperative complications such as intestinal obstruction, perforation, fistulas, and anastomotic complications (stenosis and dehiscence). CT examination is crucial for the detection of fluid collections, abscesses, and fistulas because it can serve as a guide of drainage and during follow-up of patients with Gardner syndrome can be used to investigate all possible sites in which desmoids might arise in addition to their relation to the graft. Because patients with transplanted bowel are generally rather a young population of reproductive age and because of technologic advances, MR may represent an effective method that does not use ionizing radiation and can therefore substitute for traditional radiologic evaluation. US represents a quick examination technique that is easily available and well tolerated by patients, and it has a role to play in the follow up of transplanted patients and in the identification of major postoperative complications. However, its role in monitoring possible rejection remains to be defined with studies on wider and more representative samples. PMID- 15759205 TI - Pancreatic head cancer: accuracy of CT in determination of resectability. AB - BACKGROUND: Pancreatic cancer is a devastating disease whose early detection remains difficult. There is no 100% reliable imaging test to diagnose and stage pancreatic cancer. We assessed the surgical value of contrast-enhanced spiral computed tomography (CT) in predicting the resectability and survival rates of patients who had pancreatic head cancer. METHODS: Eighty-nine patients who had pancreatic head cancer were investigated with spiral CT. Based on the preoperative CT results, we assigned patients to one of three CT groups based on resectability. RESULTS: A correlation between classification of CT resectability and intraoperative finding was found in 83% of patients. The sensitivity, specificity, negative predictive value, positive predictive value, and accuracy of spiral CT in identifying predictive unresectability were 79%, 82%, 91%, 62%, and 81%, respectively. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of CT in diagnosis of vascular invasion were 94%, 84.2%, 94%, 84%, and 91.3%, respectively. CONCLUSION: The use of CT in the evaluation of pancreatic tumors provides valuable preoperative assessment of surgical resectability and should be performed for clinical examination. Classifying patients by tumor resectability on CT helps to estimate more precisely the tumor stage and to prognosticate survival rates of these patients. PMID- 15759206 TI - Ruptured abdominal aortic aneurysm: endovascular treatment. AB - BACKGROUND: This report describes our preliminary experience in endovascular management of 25 ruptured abdominal aortic aneurysms (rAAAs). METHODS: In the past 3 years we treated 46 patients who had rAAA, and 25 (54.3%) were treated with an endovascular approach. Patients' mean age was 76 +/- 9 years. The diagnosis was confirmed by computed tomographic angiography in 23 patients (92%). Mean aneurysm diameter was 73 +/- 17 mm. We used an infrarenal bifurcated device in 17 patients (68%), a suprarenal bifurcated in four patients (16%), and an aortomonoiliac graft in four patients (16%). Overall, nine patients (36%) required intensive care. Every patient underwent radiologic follow-up according to the Eurostar register, with concomitant evaluation of the D-dimer level (cut off <200 microg/L) as a biological marker for endoleaks. RESULTS: The primary technical success rate was 100%. Overall in-hospital mortality rate was 20%. Mean hospitalization was 7 days (range, 3-30), and mean follow-up was 7 months. One occlusion (4%) of the iliac limb and two type II endoleaks (8%) occurred. The mean D-dimer level in type I endoleak was 1045 microg/L (range, 459-2021). CONCLUSIONS: In our experience, endovascular management of rAAA is feasible and safe and produces better results than conventional surgery, provided the morphology is suitable and the procedure is carried out by an experienced endovascular team. PMID- 15759207 TI - Radiofrequency ablation of neuroendocrine liver metastases: the Middlesex experience. AB - BACKGROUND: Current treatment options for neuroendocrine liver metastases are not widely applicable or not that effective. Image-guided thermal ablation offers the possibility of a minimally invasive, albeit palliative, treatment that decreases tumor volume, preserves most of the normal liver, and can be repeated several times. We report our experience with image-guided thermal ablation in 25 patients with unresectable liver metastases. METHODS: Since 1990 we have treated 189 tumors at 66 treatment sessions in 25 patients (12 female, 13 male; median age, 56 years; age range, 26--78 years). Thirty treatments were performed with a solid state laser, and 36 treatments were performed with radiofrequency ablation. All but one treatment was performed percutaneously under image guidance. Sixteen patients had metastases from carcinoid primaries, three from gastrinoma, two from insulinoma, and four from miscellaneous causes. Fourteen of 25 had symptoms from hormone secretion. RESULTS: Imaging follow-up was available in 19 patients at a median of 21 months (range, 4--75 months). There was a complete response in six patients, a partial response in seven, and stable disease in one; hence, tumor load was controlled in 14 of 19 patients (74%). Relief of hormone-related symptoms was achieved in nine of 14 patients (69%). The median survival period from the diagnosis of liver metastases was 53 months. One patient with end-stage cardiac disease died after a carcinoid crisis. There were eight (12%) complications: five local and three distant, four major and four minor. CONCLUSIONS: As a minimally invasive, readily repeatable procedure that can be used to ablate small tumors, preferably before patients become severely symptomatic, radiofrequency ablation can provide effective control of liver tumor volume in most patients over many years. PMID- 15759208 TI - Radiofrequency ablation of colorectal liver metastases. AB - BACKGROUND: Untreated patients with colorectal liver metastases rarely survive 3 years, and the 3-year survival rate for patients treated with chemotherapy is 3%. The best survival rates are for the small subgroup that has operable disease, i.e., 39% at 5 years. Radiofrequency ablation (RFA) offers a new opportunity to destroy liver metastases in patients who are not surgical candidates because of disease distribution or comorbidity. METHODS: Acceptance criteria were a maximum of four or five liver lesions with a maximum diameter of 4 or 5 cm and no evidence of active extrahepatic disease. Nearly all treatments were performed percutaneously using ultrasound, computed tomography, or magnetic resonance imaging (or some combination) for guidance and monitoring. RFA is a minimally invasive procedure that can be readily repeated. General anesthesia facilitates the procedure but is not essential. Multiple overlapping ablations are required to ensure optimal treatment in all but the smallest tumors. RESULTS: In our cohort of 167 patients with colorectal liver metastases, 73 fulfilled the optimal acceptance criteria (5 or fewer tumors that were /=75% success rate) on 21 skills (42%). Both groups experienced difficulties (/=30%; F=19.05, P <.001) and a significant interaction between initial migration percentage and initial migration percentage age (F=7.5, P <.01) was noted. Initial migration percentage age (24 mo) was not significant (F=.95, P =.34). Patients who had an initial migration percentage of 30% or more and were less than 24 months old were more likely to have a decrease in migration percentage after BTX A injection compared with patients who were older than 24 months and who had an initial migration percentage of 30% or more. CONCLUSIONS: Improvement in hip migration percentage after BTX-A injection is a function of age and the initial migration percentage. BTX-A injections to adductor muscles may be beneficial for some children with CP. PMID- 15759225 TI - Muscle sympathetic nerve activity during isometric exercise in patients with cerebrovascular accidents. AB - OBJECTIVES: To define isometric exercise-induced pressor responses in patients with cerebrovascular accidents (CVAs) and to assess potential cardiovascular and sympathetic nervous system abnormalities during isometric exercise in CVA. DESIGN: Nonrandomized study. SETTING: University laboratory setting. PARTICIPANTS: Eight men with CVA who had documented damage of subcortical structures and 8 sex-matched controls. INTERVENTIONS: A 2-minute sustained contraction of elbow flexor muscles in the unaffected side at 35% of maximal voluntary contraction (MVC; isometric exercise). MAIN OUTCOME MEASURES: Heart rate, arterial blood pressure, and muscle sympathetic nerve activity (MSNA), recorded from the peroneal nerve on the affected side. RESULTS: The percent changes in total MSNA, heart rate, and mean blood pressure in patients with CVA increased during isometric exercise but were attenuated compared with the controls. Total MSNA (mean burst amplitude per minute times burst rate) increased significantly in CVA and control subjects during isometric exercise by 18.7%+/ 6.3% and 95.8%+/-25.2%, respectively. CONCLUSIONS: The attenuated pressor responses during isometric exercise in subjects with CVA relative to the controls indicated damage to subcortical structures; such damage lowered sympathetic nervous response to isometric exercise. Our findings suggest that isometric exercise at 35% of MVC does not put patients with CVA at risk for serious tachycardia or hypertension. PMID- 15759226 TI - Skilled nursing facility rehabilitation and discharge to home after stroke. AB - OBJECTIVE: To determine the relation between rehabilitation therapy (RT) intensity and time to discharge home for stroke patients in skilled nursing facilities (SNFs). DESIGN: Retrospective cohort study. We used regression analyses, stratified by expected outcome, and propensity score adjustment. Setting All SNFs in Ohio, Michigan, and Ontario, Canada. PARTICIPANTS: A cohort of residents, aged 65 and over, admitted from hospitals to SNFs with a diagnosis of stroke (N=23,824). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Time to discharge home from an SNF. RESULTS: RT was given to more than 95% of residents for whom discharge was expected within 90 days and to more than 60% of residents for whom discharge was uncertain or not expected. RT increased the likelihood of discharge to the community for all groups except those expected to be discharged within 30 days. The dose-response relation was strongest for residents with either an uncertain discharge prognosis or no discharge expected. CONCLUSIONS: Postacute residents with an uncertain prognosis are an important target population for intensive RT. PMID- 15759227 TI - Rehabilitation outcomes of 5 patients with severe West Nile virus infection: a case series. AB - OBJECTIVE: To report inpatient rehabilitation outcome in severe cases of West Nile virus (WNV) infection. DESIGN: Retrospective case series. SETTING: Freestanding rehabilitation hospital. PARTICIPANTS: Five consecutive patients admitted to an inpatient rehabilitation hospital with proven WNV infection. PATIENTS: had severe neurologic manifestations and functional deficits. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: FIM instrument scores. RESULTS: On admission, overall FIM scores ranged from 31 to 68, with a mean of 44.40+/-15.85; overall FIM scores ranged from 52 to 90, with a mean of 76.00+/ 15.03 at discharge. CONCLUSIONS: All patients demonstrated significant functional improvement ( P <.001). Length of stay ranged from 10 to 71 days, with a mean of 32 days. Lower functional outcome and higher cost of care were noted in patients and was attributable to severe muscle weakness and axonal neuropathy. PMID- 15759228 TI - Predictors of outcome in prolonged posttraumatic disorders of consciousness and assessment of medication effects: A multicenter study. AB - OBJECTIVES: To develop predictive models of recovery from the vegetative state (VS) and minimally conscious state (MCS) after traumatic brain injury (TBI) and to gather preliminary evidence on the impact of various psychotropic medications on the recovery process to support future randomized controlled trials. Design Longitudinal observational cohort design, in which demographic information, injury and acute care history, neuroimaging data, and an initial Disability Rating Scale (DRS) score were collected at the time of study enrollment. Weekly follow-up data, consisting of DRS score, current psychoactive medications, and medical complications, were gathered until discharge from inpatient rehabilitation. SETTING: Seven acute inpatient rehabilitation facilities in the United States and Europe with specialized programs for treating patients in the VS and MCS. PARTICIPANTS: People with TBI (N=124) who were in the VS or MCS 4 to 16 weeks after injury. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: DRS score at 16 weeks after injury and time until commands were first followed (among those participants demonstrating no command following at study enrollment). Results DRS score at enrollment, time between injury and enrollment, and rate of DRS change during the first 2 weeks of poststudy observation were all highly predictive of both outcomes. No variables related to injury characteristics or lesions on neuroimaging were significant predictors. Of the psychoactive medications, amantadine hydrochloride was associated with greater recovery and dantrolene sodium was associated with less recovery, in terms of the DRS score at 16 weeks but not the time until commands were followed. More detailed analysis of the timing of functional improvement, with respect to the initiation of amantadine provided suggestive, but not definitive, evidence of the drug's causal role. CONCLUSIONS: These findings show the feasibility of improving outcome prediction from the VS and MCS using readily available clinical variables and provide suggestive evidence for the effects of amantadine and dantrolene, but these results require confirmation through randomized controlled trials. PMID- 15759229 TI - Traumatic brain injury as a relevant cause of growth hormone deficiency in adults: A KIMS-based study. AB - OBJECTIVES: To characterize further the clinical manifestations and the efficacy of growth hormone (GH) replacement therapy in patients with adult-onset growth hormone deficiency (GHD) reported in the KIMS (Pfizer's international metabolic database) as caused by traumatic brain injury (TBI) and to compare them with nonirradiated patients whose GHD was due to a nonfunctioning pituitary adenoma (NFPA). DESIGN: Observational study. SETTING: Subjects selected from the KIMS database. PARTICIPANTS: Fifty-one patients with GHD resulting from TBI and 688 patients with GHD resulting from NFPA. Both groups were selected from the KIMS and had adult-onset GHD with GH replacement therapy only after KIMS entry and before and after KIMS entry. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Age, body mass index, age at disease onset, age at disease diagnosis, age at KIMS entry, final height, GH peak at testing, GH replacement dose, routine biochemical analysis, clinical manifestations of disease, and quality of life measurements. RESULTS: Patients with TBI were significantly younger at study entry and were younger both at pituitary disease onset and at GHD diagnosis, but they showed a significant delay in treatment. When comparing patients not treated with GH before entering in the KIMS, patients with TBI were significantly shorter (167.2+/-1.7 cm) than those with NFPA (171.6+/-0.4 cm) in final height. TBI patients had lower GH reserves than NFPA patients, and although the latter group experienced more positive changes, both groups benefited from GH replacement therapy. CONCLUSIONS: Patients with GHD due to TBI showed a significant reduction in height and a reduction in pituitary GH reserve and were diagnosed and treated with inappropriate delay. PMID- 15759230 TI - A 2-item screen for depression in rehabilitation inpatients. AB - OBJECTIVE: To generate a brief screening measure to assist frontline rehabilitation health care workers to rapidly and validly screen patients for depressive symptoms. DESIGN: Case series. SETTING: Inpatient rehabilitation unit. PARTICIPANTS: Five hundred patients (age, >/=60y) cognitively able to complete the outcome measure. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Geriatric Depression Scale (GDS). RESULTS: Based on the strength of the correlation with the GDS total score, 5 items were selected for model development. A 2-item screen, consisting of items "downhearted and blue" and "worthless," correctly classified 78.4% of patients, with only 3.6% of patients having false-negative classifications. The 2-item screen did not differ significantly in its accuracy or predictive utility from the more inclusive models. CONCLUSIONS: The identified 2-item screening instrument may be an effective tool for health care staffs to use in their efforts to identify symptoms of depression in older adults on a rehabilitation unit. PMID- 15759231 TI - Characteristics of depressed and nondepressed women with physical disabilities. AB - OBJECTIVES: To describe the characteristics of depressed versus nondepressed women with disabilities. DESIGN: Survey. SETTING: Women were recruited through private and public health clinics and various community organizations. PARTICIPANTS: A sample of 443 predominantly ethnic-minority women, ages 18 to 83 years, with physical disabilities. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Beck Depression Inventory-II (BDI-II). RESULTS: Approximately half (51%) of the sample had scores in the mildly depressed range or higher on the BDI-II. Women classified as depressed were more likely to have multiple sclerosis, to be younger, or to have a shorter duration of disability. Only 44% of the women with scores exceeding the BDI-II cutoff for significant depressive symptomatology had received recent treatment for depression, with Hispanic women being the least likely to report receiving treatment. Depressed women reported significantly more health conditions, reflecting common symptoms of depression, conditions linked with depression in the general population, conditions known to be secondary to the primary condition, and conditions known to be common side effects of medications. Depression was not related to functional limitations. CONCLUSIONS: Depression is a serious mental health problem facing many women with physical disabilities that does not appear to be adequately addressed by health care professionals. PMID- 15759232 TI - Reamputation, mortality, and health care costs among persons with dysvascular lower-limb amputations. AB - OBJECTIVES: To examine 12-month reamputation and mortality rates as well as acute and postacute medical care costs among a large cohort of persons with dysvascular amputations. DESIGN: Retrospective cohort study. SETTING: General community. PARTICIPANTS: Medicare beneficiaries identified from the Centers for Medicare and Medicaid Services data as undergoing a lower-limb amputation secondary to vascular disease in 1996. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Twelve-month reamputation and mortality rates, and acute and postacute medical care costs, by initial amputation level and presence or absence of diabetes. RESULTS: A total of 3565 persons, corresponding to 71,300 Medicare beneficiaries nationwide, were identified from the claims data as undergoing lower-limb amputations in 1996. Twenty-six percent of them required subsequent amputation procedures within 12 months, and more than one third died within 1 year of their index amputation. Acute and postacute medical care costs associated with caring for beneficiaries with a dysvascular amputation exceeded $4.3 billion yearly. There were marked differences in patient characteristics, progression of amputation to higher levels, service use, and mortality among dysvascular amputees with and without a comorbidity of diabetes. Diabetic amputees were younger than those without diabetes; they were also more likely to be men, to have more comorbidities, and to have undergone their first amputation at an earlier age than persons with dysvascular amputations who did not have diabetes. Although diabetic amputees were less likely to die within 12 months of the index amputation, they died at a significantly younger age than their nondiabetic counterparts. Progression to a higher level of limb loss occurred most frequently (34.5%) among persons with an initial foot or ankle amputation. Diabetic amputees were more likely than nondiabetic amputees to experience progression to a higher amputation level for all initial amputation levels. CONCLUSIONS: This study provides information that can be used by physicians when counseling patients about expected outcomes of dysvascular amputations at different levels. PMID- 15759233 TI - The prevalence of knee pain and symptomatic knee osteoarthritis among veteran traumatic amputees and nonamputees. AB - OBJECTIVE: To determine whether amputees have an increased risk of knee pain or symptomatic osteoarthritis (OA) compared with nonamputees. DESIGN: Retrospective cohort study. SETTING: Veterans Administration Patient Treatment and Outpatient Care files. PARTICIPANTS: All male unilateral (transtibial or transfemoral) traumatic amputee patients and a random sample of male nonamputees. Patients were excluded if they were younger than 40 years, had sustained a significant injury to their knee(s), or had a rheumatic disease. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The prevalence of knee pain and symptomatic knee OA. RESULTS: The age and average weight-adjusted prevalence ratio of knee pain among transtibial amputees, compared with nonamputees, was 1.3 (95% confidence interval [CI], 0.7-2.1) for the knee of the intact limb and 0.2 (95% CI, .05-0.7) for the knee of the amputated limb. The standardized prevalence ratio of knee pain in the intact limb and symptomatic OA among transfemoral amputees, compared with nonamputees, was 3.3 (95% CI, 1.5-6.3) and 1.3 (95% CI, 0.2-4.8), respectively. CONCLUSIONS: Stresses on the contralateral knee of amputees may contribute to secondary disability. Possible explanations include gait abnormalities, increased physiologic loads on the knee of the intact limb, and the hopping and stumbling behavior common in many younger amputees. PMID- 15759234 TI - Pruritus after intrathecal baclofen withdrawal: A retrospective study. AB - OBJECTIVES: To determine the frequency of pruritus after intrathecal baclofen (ITB) withdrawal and to study the pathophysiology of this symptom. DESIGN: Retrospective cohort study. SETTING: Rehabilitation department of a general hospital. PARTICIPANTS: Patients (N=102) implanted with an ITB pump who had been followed up since 1988. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Incidence of pruritus after withdrawal. We studied the relation between pruritus and daily dose, concentration and mode of infusion of baclofen, and cause of the central nervous system lesion inducing spasticity. RESULTS: Pruritus was observed in 10 of 23 cases of ITB withdrawal. It never occurred during the first 3 months after pump implantation. It seems likely that the segmental spinal action of baclofen is responsible for pruritus. There was no statistically significant difference between patients with ITB deprivation who did and did not experience pruritus in their daily infused dosage or in concentration and mode of infusion. Surprisingly, no pruritus was observed in patients with multiple sclerosis. CONCLUSIONS: Pruritus is a frequent symptom after ITB withdrawal. Its occurrence is probably subsequent to chronic blocking of the liberation of substance P by baclofen at the spinal level. This symptom is a good clinical predictor of baclofen withdrawal, in contrast to an isolated increase of spasticity that may be due to drug tolerance or irritant factors. Pruritus requires investigation of a possible dysfunction of the infusion system. PMID- 15759235 TI - Fracture threshold in the femur and tibia of people with spinal cord injury as determined by peripheral quantitative computed tomography. AB - OBJECTIVE: To determine bone traits of the femur and tibia with peripheral quantitative computed tomography (pQCT) that best distinguish between spinal cord injury (SCI) subjects with and without fractures. DESIGN: Cross-sectional study. SETTING: In- and outpatient paraplegic center in Switzerland. PARTICIPANTS: Ninety-nine motor complete SCI subjects (duration of paralysis, 2 mo-49 y), 21 of whom had sustained fractures of the femur or tibia. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects with SCI were questioned about the occurrence, location, and approximate date of fractures to their lower extremities. Trabecular and cortical bone mineral density (BMD), as well as bone geometric properties of distal epiphyses and midshafts of the femur and tibia, were measured by pQCT. RESULTS: Trabecular BMD of the femur and tibia distal epiphyses was found to distinguish best subjects with fractures from those without. Fractures occurred in subjects with trabecular BMD of less than 114 mg/cm 3 and less than 72 mg/cm 3 for the femoral and tibial distal epiphysis, respectively (corresponding to 46% and 29% of mean values of an able-bodied reference group). Approximately 50% of the subjects with chronic SCI (defined as time postinjury >5 y for femur data and >7 y for tibia data) had trabecular BMD values above the fracture threshold in the femur and about one third above the fracture threshold in the tibia. CONCLUSIONS: By using pQCT, it may be possible to identify subjects with SCI who are at risk of sustaining fractures of the femur and tibia through minor trauma. PMID- 15759236 TI - Spinal shrinkage and recovery in women with and without low back pain. AB - OBJECTIVES: To assess the behavior of the spinal column after loading in participants with and without low back pain (LBP) using spinal shrinkage measurements as a criterion and to analyze the relations among spinal shrinkage, recovery, and LBP. DESIGN: A case-control study with repeated measures. SETTING: University department with recruitment from primary care. PARTICIPANTS: Convenience sample of 31 women (15 with chronic, nondisabling LBP [LBP group]; 16 with no history of low back disorders [control group]). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Changes in body length as determined by using a stadiometer. RESULTS: The spinal shrinkage induced by the exercise was similar ( P >.05) in both groups (control group, mean +/- standard deviation of -3.99+/ 1.13 mm; LBP group, -4.16+/-1.54 mm). Differences in spinal length were found after 20 minutes in the recovery position ( P <.05). The control group was able to recover stature to a greater extent (111.2%+/-13.6%) than the LBP group ( P <.05), which was unable to regain stature beyond baseline (57.5%+/-25.1%). A negative correlation was found between regaining stature and low back pain ( r = .52, P <.05). CONCLUSIONS: Participants with LBP were unable to recover stature to the same extent as controls. We suggest that LBP is related to the diminished ability to recover rather than to the magnitude of the spinal shrinkage imposed during the task. PMID- 15759237 TI - Early versus delayed inpatient spinal cord injury rehabilitation: an Italian study. AB - OBJECTIVE: To examine what effect the injury-to-rehabilitation interval has on the outcome of spinal cord injury (SCI) rehabilitation. DESIGN: Retrospective study. SETTING: Spinal unit of a large rehabilitation hospital. PARTICIPANTS: Consecutive admissions were divided into groups according to age, sex, and American Spinal Injury Association impairment grade and neurologic level of injury. The patients were matched for these variables and divided into groups according to the interval from injury to admission into acute rehabilitation. This approach resulted in 150 patients with SCI grouped into 50 comparison subgroupings. Interventions Three comparison groups-short (<30 d), medium (31-60 d), and long (>60 d) time to admission (TTA)-were evaluated for rehabilitation outcomes. MAIN OUTCOME MEASURES: Barthel Index, Rivermead Mobility Index, Walking Index for Spinal Cord Injury, and motor scores at admission and discharge were examined. The changes and efficiencies were evaluated. RESULTS: The 3 groups were comparable for all medical and demographic characteristics as well as neurologic recovery. The 3 subgroups differed significantly in activity of daily living outcomes, with the short TTA group exhibiting higher Barthel Index raw discharge scores, score increases, and score efficiencies. CONCLUSIONS: Early rehabilitation seems to be a relevant prognostic factor of functional outcome. Rehabilitation intervention in patients with SCI should begin as soon as possible, in a specialized setting, because delay may adversely affect functional recovery. PMID- 15759238 TI - Carotid artery flow volume and velocity by duplex sonography in men with chronic low thoracic and lumbar spinal cord injury. AB - OBJECTIVE: To investigate the common carotid blood flow volume and velocity in men with chronic low thoracic and lumbar spinal cord injury (SCI), using duplex sonography. DESIGN: Experimental control study. SETTING: University laboratory in a department of rehabilitation medicine. PARTICIPANTS: Twenty men with SCI between the T7 and L1 segments and 15 age- and sex-matched control subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Common carotid artery flow volume and velocity. RESULTS: The mean carotid artery time-averaged flow velocity of subjects with SCI was significantly lower than that of control subjects. Moreover, the mean carotid flow volume of subjects with SCI was significantly lower than that of control subjects (right side: SCI, .52+/-.14 L/min; controls, .67+/-.10 L/min; P <.05; left side: SCI, .54+/-.15 L/min; controls, .72+/-.14 L/min; P <.05). CONCLUSIONS: There are reduced common carotid artery flow volumes and flow velocities in men with chronic SCI. The reduction may result from disturbances of blood distribution induced by reduced vasoconstriction tone below the level of SCI lesion. PMID- 15759239 TI - The logical choice of muscles for the needle-electromyography evaluation of cervical radiculopathy. AB - OBJECTIVE: To find the theoretically smallest subsets of muscles for needle electromyography screening of cervical radiculopathies that meet or exceed the American Association of Electrodiagnostic Medicine (AAEM) guideline: ascertaining that each root and its adjacent roots are represented by at least 2 muscles each, innervated by those roots via different peripheral nerves. DESIGN: Twenty-two and 36 muscle sets and their myotomal innervation were derived from the literature, and rearranged into 15 and 23 unique muscle groups by root and peripheral nerve innervation. All 2 circumflex 15 and 2 circumflex 23 subsets thereof were respectively identified. The best approximations that met or exceeded the AAEM criteria were computed for each subset and the smallest subsets that qualified were retained. SETTING: Electromyography laboratory. PARTICIPANTS: None. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: Compliance with or exceeding the AAEM guideline in the number of muscles sampled per damaged root and the number of muscles sampled per adjacent normal roots. RESULTS: The smallest subsets satisfying or exceeding the AAEM guideline are of 8, 9, and 10 muscles each, and are enumerated in tables 3 and 4 in this article. CONCLUSIONS: Electromyographers may choose a set or sets from tables in this article that best suits them, with confidence that they comply with or exceed the AAEM guideline while causing minimal pain to their patients and using the shortest possible procedure. All the roots can be screened at once to best differentiate between normal and damaged roots, and, in most cases, to reach a final diagnosis. Nonetheless, when needed, each set may be the basis for a more extensive workup. PMID- 15759240 TI - Changes of calcitonin gene-related peptide in primary sensory neurons and their central branch after nerve root compression of the dog. AB - OBJECTIVE: To investigate changes in axonal flow after nerve root compression by using immunohistochemical techniques to detect calcitonin gene-related peptide (CGRP), which is thought to be involved in pain sensation. DESIGN: Experimental, controlled study. SETTING: University medical school in Japan. ANIMALS: Forty adult mongrel dogs (weight, 7-15kg). INTERVENTIONS: In dogs, the lumbar nerve roots were compressed using 4 types of clips with different pressures. Changes of CGRP levels in the spinal dorsal horn, dorsal root, and dorsal root ganglia (DRG) were examined immunohistochemically after compression for 24 hours or for 1 week. MAIN OUTCOME MEASURES: CGRP-positive neurons and CGRP-positive fibers. RESULTS: After compression, axonal flow in the dorsal root was impaired, accumulation of CGRP was observed distal to the site of compression, and the number of DRG cells showing positively for CGRP decreased. Compression for 1 week resulted in a decrease in the number of CGRP-positive fibers in the spinal dorsal horn. CONCLUSIONS: These findings indicate that CGRP dynamics are modified by nerve compression. PMID- 15759241 TI - Correlation among physical impairments, pain, disability, and patient satisfaction in patients with chronic neck pain. AB - OBJECTIVE: To investigate the correlations among pain, physical impairments, disability, and patient satisfaction in patients with chronic neck pain. DESIGN: A longitudinal cohort study with 6-month follow-up. SETTING: Institutional practice. PARTICIPANTS: Subjects (N=218) with chronic neck pain. INTERVENTIONS: Subjects were treated with different physiotherapy modalities. MAIN OUTCOME MEASURES: Data were obtained for self-reported disability, verbal numeric pain scale, patient satisfaction, and 2 measures of physical impairments during the initial visit, at 6 weeks, and finally at 6 months. RESULTS: The correlation among 4 sets of measurements varied. Moderate correlation was noted between disability and patient satisfaction ( r range, .50-.65), and between disability and pain ( r range, .55-.63). A fair relationship was found between pain and patient satisfaction ( r range, .43-.48), but only weak relationships were found between physical impairments and pain ( r range, -.08 to -.25). The correlations tended to increase in the follow-up assessments. CONCLUSIONS: No strong correlations were found among disability, patient satisfaction, pain, and physical impairments although the correlations tended to increase in the follow up assessments. The findings support the suggestion that clinicians should address as many relevant aspects of a presenting clinical entity as possible in the management of chronic neck pain. PMID- 15759242 TI - Continuous monitoring of upper-limb activity in a free-living environment. AB - OBJECTIVE: To develop and evaluate a system for the objective measurement of upper-limb activity during a person's activities of daily living. DESIGN: Construct validation study. SETTING: Home and work environment. PARTICIPANTS: Ten able-bodied participants and 10 stroke patients. INTERVENTIONS: A novel activity monitor was developed for monitoring upper-limb activity. The Strathclyde Upper Limb Activity Monitor (SULAM) gave a signal proportional to the vertical displacement of the wrist with respect to the shoulder. Participants wore the SULAM on both upper limbs for 8 hours while performing their normal daily activities. MAIN OUTCOME MEASURES: SULAM-derived variables quantified the total upper-limb activity and the range in which movement of the upper limb occurred. RESULTS: Data showed that the dominant arm of the able-bodied participants was 19% more active than the nondominant arm, whereas the unaffected arms of the group of stroke patients was used 3 to 6 times more than their affected arm. For all the variables, in both groups, there were significant differences between their upper limbs except for movement below midtrunk. There were also significant differences between groups for bimanual movement time ( P =.000) and composite movement time ( P =.000). CONCLUSIONS: The results suggest that the quantification of vertical position of the wrist with respect to the shoulder might reflect upper-limb activity. By using the SULAM, it is possible to monitor upper-limb activity over the course of a person's normal day, with minimal interference. This study provides preliminary evidence of the SULAM as a useful tool for objectively evaluating interventions aimed at improving upper-limb activity and function. PMID- 15759243 TI - Effect of passive stretch on reproducibility of hip range of motion measurements. AB - OBJECTIVE: To examine the intra- and intertester and intra- and interday reliability of hip passive range of motion (PROM) measurements and the effect of passive stretch on the reproducibility of PROM measurements. DESIGN: Reliability study. SETTING: Rehabilitation clinic at university hospital. PARTICIPANTS: Twenty volunteers (12 women, 8 men; age range, 18-45 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Two physical therapists made 2 PROM measurements on the first day and repeated them once after 2 days. The PROM in hip flexion, extension, and inner rotation and knee flexion were determined. The measurements of the hip flexion and inner rotation involved either 1 or 8 short term passive stretches. The reproducibility was expressed as the intraclass correlation coefficient (ICC) and the coefficient of variation (CV) (in percent). RESULTS: The passive stretch increased significantly ( P <.05 to P <.001) the PROM of the hip joint. The reproducibility of the PROM measurement did not increase after repetitive passive stretch. The intra- and interday intra- and intertester ICC and CV of the PROM of the hip flexion and hip inner rotation ranged from .655 to .988 and 2.1% to 12.6%, respectively. The intra- and intertester intra- and interday ICC of the PROM of the hip extension ranged from .740 to .961, and the CV ranged from -85.5% to 242.6%. The intra- and intertester intra- and interday ICC of the PROM of the knee flexion ranged from .497 to .913, and the CV ranged from 2.5% to 9.9%. CONCLUSIONS: Repetitive stretching increased significantly the PROM in hip flexion and inner rotation, but the reproducibility of the PROM measurement did not improve. The hip flexion and the hip inner rotation PROM measurements had moderate to very high intra- and interday intra- and intertester reliabilities. The reliability of the PROM in hip extension and knee flexion was poor. PMID- 15759244 TI - Device-assisted muscle strengthening in the rehabilitation of patients after surgically stabilized vertebral fractures. AB - OBJECTIVES: To investigate the effects of a device-assisted muscle strengthening exercise program on the surgically traumatized multifidus musculature and on the intact longissimus and iliocostal muscles and to assess the possible relationship between patients' reported pain symptoms and changes in muscle strength or changes in cross-sectional area (CSA) of the musculature. DESIGN: Open, prospective noncontrolled prepost intervention study. SETTING: University affiliated center for ambulant physiotherapy. PARTICIPANTS: Fifteen patients who had undergone dorsal osteosynthesis for treatment of thoracolumbar vertebral fracture and who complained of persistent back pain. INTERVENTION: Twelve-week device-assisted training exercise program. MAIN OUTCOME MEASURES: Patients' pain score, muscle strength, and the CSA of the paravertebral musculature determined by magnetic resonance imaging (MRI) were assessed before and after the exercise program. RESULTS: MRI findings revealed no increase in the CSA of the multifidus muscle in any patient (median change, -.27 cm 2 ). All patients, however, exhibited hypertrophy of both the longissimus and iliocostal muscles (median change, 1.39 cm 2 ). Significant increase in muscle strength was observed in 14 of 16 patients (median increase, 56%; range, 0.7%-126.4%). The median overall pain score improved from 19 (range, 7-24) to 16 (range, 5-27). The change in muscle strength and muscle CSA, however, showed no correlation. There was also no correlation between increase in muscle strength and changes in pain scores. CONCLUSIONS: The device-assisted training program resulted in hypertrophy of the iliocostal and longissimus muscles and an increase in muscle strength in patients with surgically stabilized vertebral fractures. About half of the patients reported relief of pain. No correlation was found between hypertrophy, increase in muscle strength, and relief of pain. The surgically damaged multifidus musculature, however, did not show any change in CSA and was not accessible to rehabilitative measures. PMID- 15759245 TI - Static magnetic fields neither prevent nor diminish symptoms and signs of delayed onset muscle soreness. AB - OBJECTIVE: To determine whether application of a commercially available static magnetic field would alter the signs and/or symptoms of delayed onset muscle soreness (DOMS) produced by exhaustive eccentric exercise. DESIGN: A double blinded, randomized, and placebo-controlled study, with subjects serving as their own controls. SETTING: An outpatient physical therapy and performance center. PARTICIPANTS: Twenty-three healthy volunteers (18 women; mean age, 30 y; range, 18-40 y; 5 men; mean age, 29 y; range, 19-39 y). INTERVENTION: After exhaustive eccentric exercise of both the right and left elbow flexor muscle groups, subjects received daily treatment with either a 350G magnet or a placebo device for 5 consecutive days. MAIN OUTCOME MEASURES: Outcome variables, including anthropometric measurements, perceived discomfort, and muscle force production, were compared using linear mixed models. RESULTS: Arm circumference, relaxed elbow flexion angle, and pain increased, whereas active elbow flexion angle and maximal isometric torque decreased transiently before returning to near baseline. No significant difference in outcome variables existed between the treated and control arms. Participants reported less pain in both treated and control arms after each session, suggesting a placebo effect. CONCLUSIONS: Static magnetic fields were no more effective than placebo in preventing DOMS. PMID- 15759246 TI - Measuring standing hindfoot alignment: reliability of goniometric and visual measurements. AB - OBJECTIVE: To examine the reliability of a functional weight-bearing measure of hindfoot alignment, the standing tibiocalcaneal angle (STCA), and to compare the relative reliabilities of goniometrically and visually estimated STCAs. DESIGN: Prospective blinded comparison. SETTING: Sports medicine center. PARTICIPANTS: Eighteen asymptomatic volunteer subjects (10 men, 8 women; age range, 22-41y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Two experienced examiners completed 2 blinded goniometric STCA and 2 blinded visual STCA measurements on each subject's right and left ankles in random order. RESULTS: Quantitative visual and goniometric STCAs were similar (visual mean range, 5.61 degrees -6.50 degrees valgus vs goniometric mean range, 5.50 degrees -6.94 degrees valgus), and both measurements exhibited good to excellent intrarater reliabilities (intraclass correlation coefficient [ICC] range, .80-.94; 95% prediction limits, 1.51 degrees -2.06 degrees ). Interrater ICCs were only fair for both measurement methods (.50-.75; 95% prediction limits, 2.2 degrees -4.1 degrees ). In terms of relative reliability, the visual STCA and goniometric STCA exhibited good to excellent agreement (ICC range, .64-.95). CONCLUSIONS: The STCA as described herein exhibited acceptable intrarater reliability for clinical use but may not be acceptably reliable between experienced examiners. The visual and goniometric STCA measurements were quantitatively similar and exhibited similar reliability. Using either method, changes of up to 2 degrees over time may be attributable to measurement error. Clinicians may consider using either STCA measurement in evaluating patients with lower-limb injuries or during screening of high-risk populations. PMID- 15759247 TI - Vitamin K deficiency and osteopenia in elderly women with Alzheimer's disease. AB - OBJECTIVE: To analyze the relation between vitamin K status and bone mineral density (BMD) in women with Alzheimer's disease (AD). DESIGN: Cross-sectional study. SETTING: Outpatient departments of neurology and neuropsychiatry at a hospital in Japan. Participants One hundred women with AD (mean age, 79.8 y) and 100 age-matched community dwelling controls (mean age, 80.6 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Patients were divided into 2 groups according to the degree of dementia: the mild AD group was composed of patients with a score in Mini-Mental State Examination (MMSE) of 16 and above (n=42); patients in the severe AD group had MMSE scores below 15 (n=58). We assessed body mass index (BMI). BMD was measured by computed x-ray densitometry. Serum concentrations of vitamin K 1 , 25-hydroxyvitamin D (25[OH]D 3 ), intact parathyroid hormone (PTH), and Glu osteocalcin (OC) were measured. RESULTS: BMI was significantly lower in women with more severe AD. Metacarpal BMD ( P <.02) and serum concentrations of vitamin K 1 ( P <.03) and 25(OH)D 3 ( P <.001) were significantly lower in the severe AD group than in the mild AD group. Serum levels of intact PTH and Glu OC in severely demented patients were higher than those with mild dementia ( P <.001). Serum PTH concentration correlated negatively with serum 25(OH)D 3 level ( r =-.241, P =.016). Serum concentration of vitamin K 1 correlated positively with that of 25(OH)D 3 ( r =.423, P <.001) and MMSE score ( r =.353, P <.001), and negatively with Glu OC ( r =-.580, P <.001). CONCLUSIONS: In female AD patients, nutritional vitamin K 1 deficiency may reduce production of fully carboxylated OC, which in turn may cause reduced BMD. Lower BMIs in more severe AD may imply the presence of general malnutrition in such a patient group. PMID- 15759248 TI - Osteoporosis risk and low bone mineral density in women with physical disabilities. AB - OBJECTIVES: To describe the results of bone mineral density (BMD) screening in women with different physical disabilities and to identify their risk factors for osteoporosis and use of strategies to prevent osteoporosis. DESIGN: A cross sectional descriptive study. SETTING: Community. PARTICIPANTS: Convenience sample of 429 community-residing women with different disabilities. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Peripheral BMD screening of the os calcis, using dual-energy x-ray absorptiometry. A self-administered survey addressed osteoporosis risk factors, use of osteoporosis prevention strategies, and previous recommendations for BMD screening or testing. RESULTS: Based on World Health Organization criteria, 30.5% and 22.6% of the total sample had BMD values indicative of osteopenia and osteoporosis, respectively. The number of self reported osteoporosis risk factors ranged from 0 to 10 (mean, 4.5+/-1.7). Subjects reported using few strategies to reduce their osteoporosis risks. One quarter of the women reported previous BMD screening or testing; about one third reported receiving recommendations for testing or screening by a health care provider. CONCLUSIONS: More than half of the sample had low BMD, and it was present in premenopausal as well as menopausal women. Despite low BMD and the presence of osteoporosis risk factors, less than a third of the women previously had BMD testing or screening or recommendations for such testing. Because low BMD is common in women with disabilities, greater attention must be directed toward their risk for osteoporosis and osteoporotic fractures. Prevention strategies to reduce osteoporosis risk should be considered, although their efficacy is not yet known in populations with disabilities. PMID- 15759249 TI - Olanzapine for the treatment of hemiballismus: A case report. AB - Hemiballismus is a rare movement disorder characterized by involuntary, large amplitude movements of the limbs of 1 side of the body. We describe the case of a man in his late sixties with slurred speech, agitation, and right-sided hemiballismus resulting from a left thalamic hemorrhagic stroke. Treatment with haloperidol was unsuccessful, but both the hemiballismus and agitation diminished significantly after initiation of olanzapine (Zyprexa). The improvement in the hemiballismus was quantified by recording the number of hemiballistic movements that occurred while the patient performed standardized 30-minute sessions (daily for 5d). With the first task (reaching within the base of support while seated), the average number of hemiballismic movements per session decreased from a baseline of 23.5 to 3.0 in the upper extremity and from 20.5 to 7.0 in the lower extremity. With the second task (catching a ball while seated), the abnormal movements decreased from 52 to 6.3 in the upper extremity and from 34.5 to 2.7 in the lower extremity. This case suggests that olanzapine may be a valuable pharmacologic alternative for patients with hemiballismus. PMID- 15759250 TI - Intracerebral hemorrhage secondary to autonomic dysreflexia in a young person with incomplete C8 tetraplegia: A case report. AB - Intracerebral hemorrhage is an unusual complication of autonomic dysreflexia and can be fatal if massive bleeding occurs with brain herniation. We report the case of a man in his midthirties with incomplete tetraplegia who suffered right putaminal hemorrhage during an episode of autonomic dysreflexia. Prompt recognition and removal of the triggering factors of autonomic dysreflexia quickly brought his blood pressure under control and the patient had a favorable functional outcome after rehabilitation. A review of the literature suggests that the location of hemorrhage in autonomic dysreflexia-induced cases is similar to that in the general population. The most common triggering factors are bladder distension in men and labor induction in women. Hemorrhagic stroke can also occur in patients with incomplete spinal cord injury (SCI) who develop autonomic dysreflexia. The role of sympathetic skin response examination is also discussed. This life-threatening complication should be kept in mind in the case of people with SCI. PMID- 15759251 TI - Severe trismus as a complication of cerebrovascular accident: a case report. AB - Spastic hypertonia is a common sequelae after a cerebrovascular accident (CVA) and is a component of an upper motoneuron lesion. Management of spastic hypertonia may involve the use of centrally acting agents, peripheral blockade, and ablative therapies. We report a case of spastic hypertonia leading to severe trismus after CVA that was successfully treated with botulinum toxin type A. Severe trismus is a potentially life-threatening condition that may lead to permanent functional impairment if not promptly diagnosed and treated. Using electromyography for botulinum toxin muscle selection in this case appears to have helped decrease the dose of botulinum toxin given as well as minimize potential side effects. PMID- 15759252 TI - A comparison of spinergy versus standard steel-spoke wheelchair wheels. AB - OBJECTIVES: To compare the energy efficiency of straight-line wheeling using Spinergy wheels as compared with standard steel-spoke wheels, and to assess the 2 wheels in terms of user comfort and wheeling preference during a wheeling course with multiple turns and surfaces. DESIGN: Nonblinded randomized crossover trial. SETTING: Rehabilitation center. PARTICIPANTS: Twenty persons with paraplegia (neurologic level T6 and below). INTERVENTION: Wheeling a straight line and obstacle course with Spinergy or standard spoke wheelchair wheels. MAIN OUTCOME MEASURES: Velocity and Physiological Cost Index (PCI) while wheeling over ground at a self-selected pace, and the User Preference Questionnaire after wheeling an obstacle course, using Spinergy or standard spoke wheelchair wheels. RESULTS: There was no significant difference in wheeling energy efficiency between the Spinergy and the steel-spoke wheels as measured by PCI ( P =.975). When rated for overall comfort, the Spinergy wheels were preferred over steel-spoke wheels ( P =.002). CONCLUSIONS: Spinergy wheels provided a more comfortable ride, but did not differ from standard steel-spoked wheels in terms of energy efficiency. The increased comfort may have important implications in patient management of pain and spasticity. PMID- 15759253 TI - Misuse of the Pearson chi-square test of association. PMID- 15759254 TI - Transfection efficiency and uptake process of polyplexes in human lung endothelial cells: a comparative study in non-polarized and polarized cells. AB - BACKGROUND: Following systemic administration, polyplexes must cross the endothelium barrier to deliver genes to the target cells underneath. To design an efficient gene delivery system into lung epithelium, we evaluated capture and transfection efficiencies of DNA complexed with either Jet-PEI (PEI-polyplexes) or histidylated polylysine (His-polyplexes) in human lung microvascular endothelial cells (HLMEC) and tracheal epithelial cells. METHODS: After optimizing growth conditions to obtain a tight HLMEC monolayer, we characterized uptake of polyplexes by flow cytometry and evaluated their transfection efficiency. Polyplexes were formulated as small particles. YOYO-labelled plasmid fluorescence intensity and luciferase activity were used as readouts for uptake and gene expression, respectively. RESULTS: PEI-polyplexes were more efficiently taken up than His-polyplexes by both non-polarized (2-fold) and polarized HLMEC (10-fold). They were mainly internalized by a clathrin-dependent pathway whatever the cell state. In non-polarized cells, His-polyplexes entered also mainly via a clathrin-dependent pathway but with an involvement of cholesterol. The cell polarization decreased this way and a clathrin-independent pathway became predominant. PEI-polyplexes transfected more efficiently HLMEC than His polyplexes (10(7) vs. 10(5) relative light units (RLU)/mg of proteins) with a more pronounced difference in polarized cells. In contrast, no negative effect of the cell polarization was observed with tracheal epithelial cells in which both polyplexes had comparable efficiency. CONCLUSIONS: We show that the efficiency of polyplex uptake by HLMEC and their internalization mechanism are polymer dependent. By contrast with His-polyplexes, the HLMEC polarization has little influence on the uptake process and on the transfection efficiency of PEI polyplexes. PMID- 15759256 TI - Reducing the glucose uptake rate in Escherichia coli affects growth rate but not protein production. AB - Although glucose is an inexpensive substrate widely used as a carbon source in Escherichia coli recombinant fermentation technology, 10-30% of the carbon supply is wasted by excreting acetate. In addition to the loss of carbon source, the excretion of a weak acid may result in increased energetic demands and hence a decreased yield. Because glucose can enter the cell via several transport systems, isogenic strains defective in one or two of these transport systems were constructed. The effects of changes in the glucose uptake capacity on the in vivo flux distribution to a desired end product (beta-galactosidase) and to acetate were studied. The lack of one of the components (IICB(Glc) protein) of the glucose-phosphoenolpyruvate phosphotransferase system (Glc-PTS) reduced the growth rate significantly. The maintenance of a low-copy plasmid in this strain resulted in further arrest of the growth rate. However, beta-galactosidase production had no effect on growth rate. This strain directed more carbon into biomass and carbon dioxide, and less into acetate. Beta-galactosidase was produced in amounts not significantly different from the wild-type strain from half the amount of glucose. An explanation for the experimental results is given, making use of published results on metabolic regulation. PMID- 15759257 TI - Macro-structural effect of metal surfaces treated using computer-assisted yttrium aluminum-garnet laser scanning on bone-implant fixation. AB - Porous coatings have been applied to the surface of prosthetic devices to foster stable device fixation. The coating serves as a source of mechanical interlocking and may stimulate healthy bone growth through osseointegrated load transfer in cementless arthroplasty. Joint arthroplasty by porous-coated prostheses is one of the most common surgical treatments, and has provided painless and successful joint mobility. However, long-term success is often impaired by the loss of fixation between the prosthesis and bone. Porous-coated prostheses are associated with several disadvantages, including metal debris from porous coatings (third body wear particles) and irregular micro-texture of metal surfaces. Consequently, quantitative histological analysis has been very difficult. These issues arise because the porous coating treatment is based on addition of material and is not precisely controllable. We recently developed a precisely controllable porous texture technique based on material removal by yttrium-aluminum-garnet laser. Free shapes can be applied to complex, three-dimensional hard metal surfaces using this technique. In this study, tartan check shapes made by crossing grooves and dot shapes made by forming holes were produced on titanium (Ti6A14V) or cobalt chrome (CoCr) and evaluated with computer-assisted histological analysis and measurement of bone-metal interface shear strength. Width of grooves or holes ranged from 100 to 800 mum (100, 200, 500, and 800 microm), with a depth of 500 microm. When the cylindrical porous-texture-treated metal samples (diameter, 5 mm; height, 15 mm) were implanted into a rabbit femoral condyle, bone tissue with bone trabeculae formed in the grooves and holes after 2 or 4 weeks, especially in 500-microm-wide grooves. Abundant osteoconduction was consistently observed throughout 500-microm-wide grooves in both Ti6A14V and CoCr. Speed of osteoconduction was faster in Ti6A14V than in CoCr, especially in the tartan check shape made of 500-microm-wide grooves. In pushout testing, the tartan check shape made of 500-microm-wide grooves had significantly higher bone-metal interface shear strength than the dot shape or commercial porous coating. These results indicate that the tartan check shape made of 500-microm-wide grooves on metal surfaces has potential for clinical application in artificial prosthesis design. PMID- 15759258 TI - Controlled release of heparin from polypyrrole-poly(vinyl alcohol) assembly by electrical stimulation. AB - A surface modification technique was developed for the covalent immobilization of poly(vinyl alcohol) (PVA)-heparin hydrogel onto electrically conductive polypyrrole (PPY) film with the objective of achieving controlled release of heparin. First, aldehyde groups were introduced onto PPY film through poly(ethylene glycol) monomethacrylate graft copolymerization and subsequent oxidation in acetic anhydride and dimethyl sulfoxide mixture. Then, the prepared PVA-heparin hydrogel was cast onto the PPY film and covalently immobilized to the film through the reaction between the aldehyde groups on the PPY film and the hydroxyl groups of PVA. X-ray photoelectron spectroscopy was used to characterize the surface-modified film after each stage. The strong attachment of the PVA heparin layer on the PPY film was confirmed by peel test and scanning electron microscopy. The release behavior of heparin from the substrate with and without electrical stimulation was studied and the experimental results showed that the heparin release rate from the prepared substrate using an electric current of 3.5 mA is twofold higher than that without current. PMID- 15759259 TI - Fabrication of calcium sulfate/PLLA composite for bone repair. AB - The bone-repairing composite material CS/PLLA was fabricated by mixing poly-L lactic acid (PLLA) and calcium sulfate hemihydrate (CSH). The structure of the composite was analyzed with Infrared spectroscope, X-ray diffraction, and scanning electron microscope. The results indicated that the crystal pattern of calcium sulfate was affected by the addition of PLLA. PLLA part impacted the development of calcium sulfate dihydrate (CSD) crystal by slowing the conversion from CSH to CSD, so the composites are actually composed of CSH, CSD, and PLLA. The absorbing test in vitro showed that CS/PLLA composite absorbed more slowly than pure CS, suggesting the addition of PLLA can adjust the absorption rate of CS to meet different requirements. The pH value changes of the media had similar trends for different composites during the absorbing test of CS/PLLA samples in aqueous medium, which was connected to the absorption of calcium sulfate. The absorption of calcium sulfate in a certain time left a porous PLLA scaffold that will enable cells to further grow in. The surface of CS/PLLA pellets was inoculated with human osteoblasts, and the primary results showed that the osteoblasts could attach and spread on the surface, which will stimulate our desire for further study. PMID- 15759260 TI - Retrospective family study of childhood medulloblastoma. AB - Medulloblastoma is the most common malignant central nervous system tumor of childhood and can occur sporadically or in association with inherited cancer susceptibility syndromes such as the nevoid basal cell carcinoma syndrome (NBCCS). To determine whether an association existed between the risk of developing medulloblastoma and undiagnosed syndromes, we retrospectively reviewed clinical data on 33 patients with medulloblastoma from a single institution and compared them with their unaffected relatives (n = 46). Six patients had tumors showing desmoplastic histology. Two of the six met diagnostic criteria for NBCCS. One NBCCS patient had a missense mutation of patched-1 (PTCH1); the other had no identifiable PTCH1 mutation. Two patients with isolated desmoplastic medulloblastoma had an insertion and splice site mutation, respectively, in suppressor of fused (SUFU). All patients with nondesmoplastic medulloblastoma histology received molecular testing for SUFU. None of these patients had an identifiable mutation in PTCH1 or SUFU. We performed a clinical evaluation for Greig cephalopolysyndactyly syndrome (GCPS) in four medulloblastoma families, who exhibited macrocephaly as the only finding consistent with the diagnosis of GCPS. Molecular analysis of GLI3 in these four families was negative. There was a paucity of clinical findings among the majority of medulloblastoma patients in this study group to suggest a definable cancer genetic syndrome. We conclude that clinically recognizable syndromes are uncommon among patients with medulloblastoma, however, PTCH1 and SUFU mutations are present at a low but significant frequency. PMID- 15759261 TI - Construction and characterization of pta gene-deleted mutant of Clostridium tyrobutyricum for enhanced butyric acid fermentation. AB - Clostridium tyrobutyricum ATCC 25755 is an acidogenic bacterium, producing butyrate and acetate as its main fermentation products. In order to decrease acetate and increase butyrate production, integrational mutagenesis was used to disrupt the gene associated with the acetate formation pathway in C. tyrobutyricum. A nonreplicative integrational plasmid containing the phosphotransacetylase gene (pta) fragment cloned from C. tyrobutyricum by using degenerate primers and an erythromycin resistance cassette were constructed and introduced into C. tyrobutyricum by electroporation. Integration of the plasmid into the homologous region on the chromosome inactivated the target pta gene and produced the pta-deleted mutant (PTA-Em), which was confirmed by Southern hybridization. SDS-PAGE and two-dimensional protein electrophoresis results indicated that protein expression was changed in the mutant. Enzyme activity assays using the cell lysate showed that the activities of PTA and acetate kinase (AK) in the mutant were reduced by more than 60% for PTA and 80% for AK. The mutant grew more slowly in batch fermentation with glucose as the substrate but produced 15% more butyrate and 14% less acetate as compared to the wild-type strain. Its butyrate productivity was approximately 2-fold higher than the wild type strain. Moreover, the mutant showed much higher tolerance to butyrate inhibition, and the final butyrate concentration was improved by 68%. However, inactivation of pta gene did not completely eliminate acetate production in the fermentation, suggesting the existence of other enzymes (or pathways) also leading to acetate formation. This is the first-reported genetic engineering study demonstrating the feasibility of using a gene-inactivation technique to manipulate the acetic acid formation pathway in C. tyrobutyricum in order to improve butyric acid production from glucose. PMID- 15759262 TI - Down syndrome and comorbid autism-spectrum disorder: characterization using the aberrant behavior checklist. AB - To report on the cognitive and behavioral attributes of 61 children with Down syndrome (DS) and autistic-spectrum disorder (ASD) according to DSM-IV criteria; to determine the utility of the aberrant behavior checklist (ABC) to characterize these subjects for research purposes; and to test the hypothesis that subjects with DS + ASD could be distinguished from their typical DS peers using the ABC. Cross-sectional design. Cases with DS + ASD (N = 61), comparison group of DS + stereotypy movement disorder (SMD) (N = 26) and typical DS controls without behavior problems (N = 44) were ascertained and enrolled sequentially upon presentation to a DS clinic at an academic medical center over a 10-year period from 1991 to 2001. All subjects underwent neurodevelopmental and medical evaluation, and standardized cognitive testing. The parents provided responses to standardized behavioral questionnaires. Cognitive function (IQ) differed markedly across the three groups. The Lethary and Stereotypy subscales of the ABC were highly significant (P < 0.001) in distinguishing the three groups from one another. Within the ASD group differences were apparent by DSM-IV type on the Lethargy subscale, which reached significance, ANOVA (F = 0.002) and t-test (Autism > PDD, P = 0.005; PDD < CDD, P = 0.002). Using a multivariate regression model, the ABC scales alone explained 62% of variance of ASD outcome; addition of demographic variables explained up to 68% of the variance. There is good correlation between DSM-IV criteria for autism and subscales scores on the ABC in subjects with DS. This study demonstrates the feasibility of using the ABC to characterize the neurobehavioral phenotype of a cohort of children with trisomy 21 and ASD for ongoing research purposes. PMID- 15759263 TI - Karyotype/phenotype correlations in duplication 4q: evidence for a critical region within 4q27-28 for preaxial defects. PMID- 15759264 TI - Mutational analysis of the TCOF1 gene in 11 Japanese patients with Treacher Collins Syndrome and mechanism of mutagenesis. AB - Treacher Collins Syndrome (TCS) (OMIM 154500) is a congenital, craniofacial disorder inherited as an autosomal dominant trait. The responsible gene for TCS, TCOF1, was mapped to 5q32-33.1 and identified in 1996. Since then, TCOF1 mutations in patients with TCS have been reported from Europe, North and South America, however, no TCS cases from an Asian country have been molecularly characterized. Here we report mutational analysis for 11 Japanese patients with TCS for the first time, and have identified TCOF1 mutations in 9 of them. The mutations detected were various, but most likely all the mutations are predicted to result in a truncated gene product, known as treacle. One mutation frequently reported was included in our cases, but no missense mutations were detected. These findings are similar to those for the previous studies for TCS in other races. We have speculated about the molecular mechanisms of the mutations in most cases. Collectively, we have defined some of the characteristic molecular features commonly observed in TCS patients, irrespective of racial difference. PMID- 15759265 TI - Reconstructing the behaviors of extinct species: an excursion into comparative paleoneurology. AB - How can the behavior of an extinct species be reconstructed-say a dinosaur such as Allosaurus? Despite the relatively abundant fossilized remains of this and other dinosaurs, the incompleteness of the fossil record has permitted room for considerable speculation, mythology, and perhaps a bit of unsettling reflection on what factors contributed to the eventual fate of these remarkably successful animals. Among the speculations is how these 'bigger-than-life' creatures behaved, a topic that itself can attain equal diversity and grandeur. With recent advancements in measuring the relatedness of living organisms, how genetics contribute to brain development and how this relates to behavior, combined with the availability of newly discovered high quality fossils and imaging methods to exploit their secrets, novel insights into how extinct creatures such as Allosaurus intermingled with its many relatives over 100 million years ago are beginning to emerge. PMID- 15759268 TI - Proliferation and cell shape changes during ciliary body morphogenesis in the mouse. AB - Very little is known about the structure and development of the ciliary processes in the mouse eye. Our scanning electron microscope (SEM) studies reveal that, unlike other mammals, the ciliary processes form an irregular pattern, crossing over and interweaving rather than lying parallel to one another. Histological and SEM studies from embryonic day (E) 14.5 to postnatal day (P) 7 reveal that the first morphological sign of the ciliary zone is an annular bulge; this is then gradually molded to form discrete ciliary processes. The striking similarity between the developing capillary network and the adult ciliary folds suggests that the patterning template for the ciliary processes could be the underlying capillary network. Cell proliferation measurements and cell height assessments indicated that one of the first events occurring during the morphogenesis of ciliary processes is a proliferative surge around P0 in the outer ciliary epithelium. It is likely that this surge together with increasing cell heights leads to a bulging of this layer. After a slight delay, the inner ciliary epithelium responds by proliferating and extending inward toward the lens. Final shaping of the ciliary processes is achieved through cell height reductions in the inner ciliary epithelium. Thus, in the mouse, the temporal correlation between mitotic and cell height changes during ciliary body morphogenesis suggests that these processes play an integral role in the shaping of ciliary processes. PMID- 15759267 TI - Temporal and spatial expression profile of the novel armadillo-related gene, Alex2, during testicular differentiation in the mouse embryo. AB - In a screen for transcripts differentially expressed during gonadal development in mouse embryos, we identified the novel armadillo-related gene, Alex2. The armadillo (arm) family of proteins share a 42 amino acid tandem repeat motif called the arm domain, through which they interact with different binding partners. These intracellular proteins are implicated in a variety of developmental processes, including cell proliferation, migration, maintenance of tissue integrity, and tumorigenesis. Alex2 is a member of a novel subgroup within the arm family, encoding a protein with a single arm domain and a putative transmembrane or signal sequence. Alex2 has a developmentally regulated expression profile during embryogenesis in the mouse. In the urogenital system, it is strongly expressed in the developing testis but is down-regulated during ovarian development. Alex2 expression is localized within the interstitial cell lineage of the developing testis, which gives rise to peritubular myoid, endothelial, and fetal Leydig cells. Alex2 is also expressed in the developing forebrain and somites and in dorsal root ganglia. In testicular cell lines, Alex2 fusion proteins localize to membrane structures within the cell. The expression profile of Alex2 suggests that it plays a role in the development of several tissues during embryogenesis, notably testicular differentiation. In the developing testis, its expression profile suggests that Alex2 has a role in specification or development of the interstitial cell lineage. PMID- 15759269 TI - Early signals for fracture healing. AB - Fracture healing requires the cooperation of multiple molecular signaling pathways. To better understand this cascade of transcriptional events, we compared the gene expression profiles between intact bone and fractured bone at days 1, 2, and 4 using a rat femur model of bone healing. Cluster analysis identified several groups of genes with dynamic temporal expression patterns and stage-specific functions. The immediate-response genes are highlighted by binding activity, transporter activity, and energy derivation. We consider these activities as critical signals for initiation of fracture healing. The continuously increased genes are characterized by those directly involved in bone repair, thus, representing bone specific forefront workers. The constantly upregulated genes tend to regulate general cell growth and are enriched with genes that are involved in tumorigenesis, suggesting common pathways between two processes. The constantly downregulated genes predominantly involve immune response, the significance of which remains for further investigation. Knowledge acquired through this analysis of transcriptional activities at the early stage of bone healing will contribute to our understanding of fracture repair and bone related pathological conditions. PMID- 15759270 TI - Localization and regulation of phospholipase D2 by ARF6. AB - Phospholipase D (PLD) and ADP-ribosylation factor 6 (ARF6) have been implicated in vesicular trafficking and rearrangement of the actin cytoskeleton. We have explored the co-localization of rat PLD1b and rat PLD2 with wild type and mutant forms of ARF6 in HeLa cells and studied their activation by ARF6 and the role of the actin cytoskeleton. GFP-tagged PLD1 had a similar pattern to multivesicular and late endosomes and the trans-Golgi apparatus, but not to other organelles. When wild type or dominant negative ARF6 and PLD1 or PLD2 were co-expressed, they had a similar localization in cytosolic particles and at the cell periphery. In contrast, dominant active ARF6 caused cell shrinkage and had a similar localization with PLD1 and PLD2 in dense structures, containing the trans-Golgi apparatus and actin. Disruption of the actin cytoskeleton with cytochalasin D did not induce the formation of these structures. To determine, if ARF6 selectively activated PLD1 or PLD2, wild type and mutant forms of the ARF isoform were transfected together with PLD1 or PLD2. Wild type ARF6 did not affect either PLD isozyme, but dominant active ARF6 selectively activated PLD2 and dominant negative ARF6 selectively inhibited PLD2. In contrast, dominant active ARF1 or Rac1 stimulated both PLD isozymes but the ARF1 effect on PLD2 was very small. Cytochalasin D did not affect the activation of PLD by phorbol ester. The localizations of PLD and ARF6 were also analyzed by fractionation after methyl beta-cyclodextrin extraction to deplete cholesterol. The results showed that all PLD isoforms and ARF6 mutants existed in the membrane fraction, but only wild type ARF6 was dependent on the presence of cholesterol. These experiments showed that wild type ARF6 had a similar location with PLD isoforms on cell staining, but it did not colocalize with PLD isoforms in fractionation experiments. It is proposed that activated ARF6 translocates to the cholesterol independent microdomain and then activates PLD2 there. It is further concluded that PLD2 is selectively activated by ARF6 in vivo and that disruption of the actin cytoskeleton does not affect this activation. PMID- 15759271 TI - Alternative usage of Otx2 promoters during mouse development. AB - Our previous structural analysis of mouse Otx2 transcripts has revealed the existence of three different promoters and suggested that the corresponding mRNAs could exhibit specific expression patterns. Here, we analyze the precise dynamics of their expression throughout mouse development. Their spatial distribution was determined by isoform-specific in situ hybridization and their relative abundance by real-time reverse transcriptase-polymerase chain reaction. Although the three promoters may be used in the same areas, we show that transcription preferentially occurs from the proximal promoter at onset of gene activity in early embryogenesis, and switches to the more distal one in most of the sites of expression in the adult brain. During gestation, their relative utilization becomes inverted. The third promoter, which shows no activity in embryonic stem cells and is moderately expressed during embryogenesis, is mostly used in specific areas derived from the rostral part of the neural tube. PMID- 15759272 TI - Sonic hedgehog regulates proliferation of the retinal ciliary marginal zone in posthatch chicks. AB - The ciliary marginal zone (CMZ) has long been known to be a source of postembryonic neuronal production in the retinas of fish and amphibians, and more recently, birds. However, there is little known about the factors that are required for the maintenance of this neural stem cell zone. The cells of the CMZ respond to mitogens such as endothelial growth factor, insulin-like growth factor 1, and insulin, factors that are also mitogenic for embryonic retinal progenitors, suggesting that the continued expression of embryonic mitogenic factors might be required to maintain the postembryonic proliferative potential of the CMZ. To test this hypothesis, we examined the expression and functional role of a critical embryonic retinal progenitor mitogen, Sonic hedgehog (Shh) in the regulation of proliferation of the cells of the CMZ. We have found that Shh is concentrated at the retinal margin of postembryonic chicks. Moreover, we report that intraocular injection of Shh stimulates proliferation of the CMZ cells, whereas cyclopamine, an inhibitor of the Shh pathway, inhibits CMZ proliferation. We conclude that Shh signaling is an important factor in the maintenance of postembryonic retinal neurogenesis. PMID- 15759273 TI - Endothelin-1 and Neuregulin-1 convert embryonic cardiomyocytes into cells of the conduction system in the mouse. AB - The cells that form the cardiac conduction system (CCS) are recruited from embryonic cardiomyocytes. Endothelin-1 (ET-1) and Neuregulin-1 (NRG-1) have been associated with this transition in the avian and murine systems, respectively. We established murine embryonic cardiomyocyte cultures induced or not with ET-1 and/or NRG-1 to compare the expression of cardiomyocyte and CCS-specific genes. Semiquantitative reverse transcription-polymerase chain reaction analysis showed that cardiomyogenesis and CCS-specific markers, such as Nkx2.5, GATA4, Irx4, Connexin 40, Connexin 45, HF-1b, and MinK, were up-regulated in the presence of either growth factor. Additionally, immunofluorescence analysis demonstrated that ET-1 or NRG-1 increased the number of cells expressing the Purkinje fiber specific marker Connexin 40 in induced cultures but did not selectively increase their proliferation rate. Interestingly, additive effects were not observed in ET 1 and NRG-1 combination treatments. Among other possibilities, this observation suggests that these factors may interact to promote the differentiation of the murine CCS. PMID- 15759274 TI - Dynamic expression of Wnt signaling-related Dickkopf1, -2, and -3 mRNAs in the developing mouse tooth. AB - Wnt signaling is essential for tooth formation. Members of the Dickkopf (Dkk) family modulate the Wnt signaling pathway by binding to the Wnt receptor complex. Comparison of Dkk1, -2, and -3 mRNA expression during mouse tooth formation revealed that all three genes showed distinct spatiotemporally regulated expression patterns. Dkk1 was prominently expressed in the distal, incisor bearing mesenchyme area of the mandibular process during the initial stages of tooth formation. During molar morphogenesis Dkk1 was detected in the dental mesenchyme, including the preodontoblasts. Dkk2 was seen in the dental papilla, whereas Dkk3 was specifically expressed in the putative epithelial signaling centers, the primary and secondary enamel knots. Postnatally, Dkk1 was prominently expressed in the preodonto- and odontoblasts, while Dkk3 mRNAs were transiently seen in the preameloblasts before the onset of enamel matrix secretion. These results suggest that modulation of Wnt-signaling by Dkks may serve important functions in patterning of dentition as well as in crown morphogenesis and dental hard-tissue formation. PMID- 15759275 TI - Cartilage-derived morphogenetic proteins induce osteogenic gene expression in the C2C12 mesenchymal cell line. AB - Cartilage-derived morphogenetic protein-1, -2, and -3 (CDMP-1, -2, and -3) are members of the bone morphogenetic protein (BMP) family and have been shown to exhibit a variety of biological activities. In the present study, effects of these CDMPs on the temporal and spatial expression of genes in the pluripotent mesenchymal cell line C2C12 were examined. Cells cultured in the presence of CDMPs lost the characteristic elongated shape of myoblasts. At the molecular level, CDMP treatment did not change the mRNA expression of MyoD, aggrecan, Six1, and tendin. Scleraxis mRNA level was reduced by CDMP treatment. CDMP-1 and -3, but not CDMP-2, stimulated expression of osteogenic markers, such as alkaline phosphatase (AP), osteocalcin (OC), BSP, and type I collagen, in a dose- and time dependent manner. With few exceptions, the three CDMPs changed, with different potencies, the expression profile of different members of the BMP family in a similar temporal pattern. Except at the late phase of treatment, CDMP treatment did not change the expression of ActR-IA, BMPR-IA, BMPR-IB, BMPR-II, and ALK-7 mRNAs. Based on the current data, the CDMPs appear to be able to stimulate the C2C12 cells to differentiate into the osteoblast pathway. PMID- 15759276 TI - XEpac, a guanine nucleotide-exchange factor for Rap GTPase, is a novel hatching gland specific marker during the Xenopus embryogenesis. AB - cAMP is a second messenger controlling various cellular processes through cAMP dependent protein kinase (cAPK, PKA) and cyclic nucleotide-gated ion channels. Recently, the PKA-independent-cAMP-mediated signaling pathway by means of exchange protein directly activated by cAMP (Epac) has been demonstrated. Epac is a guanine nucleotide-exchange factor (GEF) for Rap, a Ras-like small GTPase. To investigate this new target for cAMP in development, we have isolated Xepac, the Xenopus laevis homologue of Epac by cDNA library screening. Xepac (Xepac1) encodes 890 amino acids, which have 57% identity with human Epac1 and 59% with that of rat Epac1 in amino acids. Whole-mount in situ hybridization and reverse transcriptase-polymerase chain reaction analysis show that XEpac is expressed both maternally and zygotically and is restricted within the developing hatching gland. Intriguingly, overexpression of XEpac induces the anterior markers XAG-1 and XOtx2 and can convert ectoderm into cement- and hatching gland-expressing cells. These results suggest that XEpac contains anterior positional information. PMID- 15759277 TI - Cell fate specification along the anterior-posterior axis of the intermediate mesoderm. AB - The vertebrate intermediate mesoderm (IM) is highly patterned along the anterior posterior (A-P) axis. In the chick embryo, the kidney tissue, which is a derivative of the IM, is generated only from IM located posterior to the sixth somite axial level, which also marks the border between cranial and trunk segments. The cellular and molecular mechanisms that govern the formation of the anterior border of the kidney morphogenetic field are currently unknown. In this study, we asked whether specific A-P patterning information is conveyed by the movement of cells through the primitive streak (PS) at different time points that consequently affects the expression of kidney genes, or by the environment that these cells encounter during their migration to the IM. In this study, we show that kidney-inductive signals are present along the whole axis, including anterior non-kidney-generating regions. These inductive signals are generated by tissues that are located medial to the anterior IM. We also demonstrate that cells that migrate through the PS of early embryonic stages (Hamburger and Hamilton stage 3-4 and earlier), which will give rise to anterior nonkidney IM, are competent to respond to these inductive factors. This prospective anterior IM tissue loses its competence to respond to kidney inducing signals during its migration from the PS to its final location in the anterior IM. We present here a model in which changes in cell competence determine the formation of the anterior border of kidney gene expression and discuss the possible evolutionary implications of this developmental mechanism. PMID- 15759278 TI - Genetic and epigenetic aspects of bladder cancer. AB - Transitional cell carcinoma of the urinary bladder has a diverse collection of biologic and functional characteristics. This is reflected in differing clinical courses. The diagnosis of bladder cancer is based on the information provided by cystoscopy, the gold standard in combination with urinary cytology findings. Many tumor markers have been evaluated for detecting and monitoring the disease in serum, bladder washes, and urinary specimens. However, none of these biomarkers reported to date has shown sufficient sensitivity and specificity for the detection of the whole spectrum of bladder cancer diseases in routine clinical practice. The limited value of established prognostic markers requires the analysis of new molecular parameters of interest in predicting the prognosis of bladder cancer patients; in particular, the high-risk patient groups at risk of progression and recurrence. Over the past decade, there has been major progress elucidating of the molecular genetic and epigenetic changes leading to the development of transitional cell carcinoma. This review focuses on the recent advances of genetic and epigenetic aspects in bladder cancer, and emphasizes how molecular biology would be likely to affect the future therapies. PMID- 15759279 TI - VP22 does not significantly enhance enzyme prodrug cancer gene therapy as a part of a VP22-HSVTk-GFP triple fusion construct. AB - BACKGROUND: VP22 is a herpes simplex virus type 1 (HSV-1) tegument protein that has been suggested to spread from cell to cell, alone or as a part of fusion proteins. Creating controversy, some reports indicate that VP22 cannot facilitate significant intercellular spreading. To study the capacity of VP22 to cause spreading and enhance thymidine kinase/ganciclovir cancer gene therapy, we constructed a novel triple fusion protein containing VP22, HSV thymidine kinase and green fluorescent protein (VP22-Tk-GFP). This fusion protein has three functional domains in the same polypeptide, thus making it possible to reliably compare the causality between transduction rate and cell killing efficiency in vitro and in vivo. METHODS: VP22-Tk-GFP was cloned into lenti- and adenoviral vectors and used for expression studies, analyses for VP22-mediated protein spreading, and to study the effect of VP22 to thymidine kinase/ganciclovir mediated cytotoxicity. The function of VP22-Tk-GFP was also investigated in vivo. RESULTS: The triple fusion protein was expressed correctly in vitro, but intercellular trafficking was not observed in any of the studied cell lines. However, under certain conditions, VP22-Tk-GFP sensitized cells more efficiently to ganciclovir than Tk-GFP. In vivo there was a trend for increased inhibition of tumor growth with VP22-Tk-GFP when ganciclovir was present, but the difference with Tk-GFP was not statistically significant. CONCLUSIONS: Based on our results, VP22 fusion proteins do not seem to traffic intercellularly at detectable levels in most tumor cell types. Even though VP22 enhanced cytotoxicity in one cell line in vitro, the effect in vivo was modest. Therefore, our results do not support the utility of VP22 as an enhancer of enzyme prodrug cancer gene therapy. PMID- 15759280 TI - Serological protein profiling of neuroblastoma by ProteinChip SELDI-TOF technology. AB - Serological proteins of neuroblastoma were profiled and analyzed by ProteinChip SELDI-TOF MS technology with five types of protein chips. By comparing with normal control, a number of protein or polypeptide signals were found significantly and consistently different in their intensities (expression levels) in tumor sera. Interestingly, nine polypeptide peaks in these proteomic features can be simultaneously detected with consistent variations by more than one type of protein chips. None of the expression differences of these nine polypeptides was found in similar comparisons between healthy controls and hepatomas. Preliminary protein identification showed hints for that some of these proteomic alterations may be closely related to the tumorigenesis of neuroblastoma. These results demonstrated the potential of serological biomarker identification for neuroblastoma by ProteinChip-SELDI technology. PMID- 15759281 TI - Salt and hypertension: a phylogenetic perspective. AB - Our hunter-gatherer ancestors appeared to survive on little salt. When today's rural dwellers move to urban environments, they increase their salt intake and the salt-sensitive among them become prone to age-related increase in blood pressure and hypertension. This paper reviews our knowledge of the mechanisms of salt disposal and plasma volume regulation, salt consumption in human evolution, salt intake and prevalence of hypertension, and the results of interventions aimed at modulating both. Finally, it discusses current hypotheses on the mechanisms of selective pressure that may have favored the emergence of a salt sensitive, hypertensive genotype. Similar to 'thrifty' genes, which supported energy savers in times of scarcity, but may now be causing obesity and type 2 diabetes, 'thirsty' genes, by acting on salt and water retention, might have helped individuals survive the challenge of volume-depleting illnesses, especially when combined with stress-inducing situations, but may now cause high BP and related damage in the post-reproductive age. PMID- 15759282 TI - Glucagon-like peptide 1(GLP-1) in biology and pathology. AB - Post-translational proteolytic processing of the preproglucagon gene in the gut results in the formation of glucagon-like peptide 1 (GLP-1). Owing to its glucose dependent insulinotropic effect, this hormone was postulated to primarily act as an incretin, i.e. to augment insulin secretion after oral glucose or meal ingestion. In addition, GLP-1 decelerates gastric emptying and suppresses glucagon secretion. Under physiological conditions, GLP-1 acts as a part of the 'ileal brake', meaning that is slows the transition of nutrients into the distal gut. Animal studies suggest a role for GLP-1 in the development and growth of the endocrine pancreas. In light of its multiple actions throughout the body, different therapeutic applications of GLP-1 are possible. Promising results have been obtained with GLP-1 in the treatment of type 2 diabetes, but its potential to reduce appetite and food intake may also allow its use for the treatment of obesity. While rapid in vivo degradation of GLP-1 has yet prevented its broad clinical use, different pharmacological approaches aiming to extend the in vivo half-life of GLP-1 or to inhibit its inactivation are currently being evaluated. Therefore, antidiabetic treatment based on GLP-1 may become available within the next years. This review will summarize the biological effects of GLP-1, characterize its role in human biology and pathology, and discuss potential clinical applications as well as current clinical studies. PMID- 15759283 TI - Combinatorial gene therapy for bone regeneration: cooperative interactions between adenovirus vectors expressing bone morphogenetic proteins 2, 4, and 7. AB - Bone morphogenetic proteins (BMPs) have demonstrated effectiveness as bone regeneration agents whether delivered as recombinant proteins or via gene therapy. Current gene therapy approaches use vectors expressing single BMPs. In contrast, multiple BMPs are coordinately expressed during bone development and fracture healing. Furthermore, BMPs likely exist in vivo as heterodimeric molecules having enhanced biological activity. In the present study, we test the hypothesis that gene therapy-based bone regeneration can be enhanced by expressing combinations of BMPs. For in vitro studies, mesenchymal cell lines were transduced with individual adenoviruses containing BMP2, 4, or 7 cDNA under control of a CMV promoter (AdBMP2, 4, 7) or virus combinations. Significantly, combined transduction with AdBMP2 plus AdBMP7 or AdBMP4 plus AdBMP7 resulted in a synergistic stimulation of osteoblast differentiation. This synergy is best explained by formation of BMP2/7 and 4/7 heterodimers. To test in vivo biological activity, fibroblasts were transduced with specific virus combinations and implanted into C57BL6 mice. Consistent with in vitro results, strong synergy was observed using combined AdBMP2/BMP7 treatment, which induced twofold to threefold more bone than would be predicted based on the activity of individual AdBMPs. These studies show that dramatic enhancement of osteogenesis can be achieved using gene therapy to express specific combinations of interacting regenerative molecules. PMID- 15759284 TI - NFAT expression in human osteoclasts. AB - Nuclear factor of activated T-cells cytoplasmic (NFATc) is a family of transcription factors originally identified in T-cells. The gene family is currently known to have four members (NFATc1 through NFATc4) which have roles both within and outside the immune system. We show that NFATc1 is the major induced NFAT in human osteoclasts, with expression greatly exceeding that of NFATc2 through NFATc4. In macrophage-like cells in culture, NFATc1 through NFATc4 are expressed at similar low levels. NFATc1 is comprised of five mRNA transcript variants known to encode three different protein isoforms. The mRNA encoding isoform C (mRNA variant 3) was the most expressed with 38 copies per nanogram followed by isoform B (mRNA variant 5) with 17 copies per nanogram of total RNA. Isoform A (mRNA variant 1) and mRNA variants 2 and 4 made up less than 1% of the total NFATc1 expressed. NFATc1 is activated by calcineurin after calcium calmodulin signalling. The induction of NFATc1 in osteoclasts was not altered in the presence of cyclosporin A, an inhibitor of calcineurin, suggesting that NFATc1 does not participate in autoregulatory activation of its own promoter. The NFATc1 variants expressed by human osteoclasts are not those normally expressed by effector T-cells but are similar to those seen in naive T-cells. PMID- 15759285 TI - Scavenging of reactive oxygen species by novel indolin-2-one and indoline-2 thione derivatives. AB - The antioxidant behavior of a series of new synthesized substituted indoline-2 ones and indolin-2-thiones was investigated in this study using an oxygen radical absorbance capacity assay (ORAC(ROO*-) and 2,2'-azobis(2-amidino-propane) dihydrochloride (AAPH) as the radical generator; system generating superoxide anion radical, O2*- (18-crown-6/KO(2)/DMSO), and the Fenton-like reaction [Co(II) + H(2)O(2) --> Co(III) + HO(*) + HO(-)]. Measurements were done using fluorescence, chemiluminescence methods, and a deoxyribose assay based on the spectrophotometry method, respectively. The results obtained indicated that the examined indoline derivatives had effective activities as radical scavengers and may be considered as an effective source for combating oxidative damage. PMID- 15759286 TI - Ultrastructure of oogenesis in the bluefin tuna, Thunnus thynnus. AB - Ovarian ultrastructure of the Atlantic bluefin tuna (Thunnus thynnus) was investigated during the reproductive season with the aim of improving our understanding of the reproductive biology in this species. The bluefin, like the other tunas, has an asynchronous mode of ovarian development; therefore, all developmental stages of the oocyte can be found in mature ovaries. The process of oocyte development can be divided into five distinct stages (formation of oocytes from oogonia, primary growth, lipid stage, vitellogenesis, and maturation). Although histological and ultrastructural features of most these stages are similar among all studied teleosts, the transitional period between primary growth and vitellogenesis exhibits interspecific morphological differences that depend on the egg physiology. Although the most remarkable feature of this stage in many teleosts is the occurrence of cortical alveoli, in the bluefin tuna, as is common in marine fishes, the predominant cytoplasmic inclusions are lipid droplets. Nests of early meiotic oocytes derive from the germinal epithelium that borders the ovarian lumen. Each oocyte in the nest becomes surrounded by extensions of prefollicle cells derived from somatic epithelial cells and these form the follicle that is located in the stromal tissue. The primary growth stage is characterized by intense RNA synthesis and the differentiation of the vitelline envelope. Secondary growth commences with the accumulation of lipid droplets in the oocyte cytoplasm (lipid stage), which is then followed by massive uptake and processing of proteins into yolk platelets (vitellogenic stage). During the maturation stage the lipid inclusions coalesce into a single oil droplet, and hydrolysis of the yolk platelets leads to the formation of a homogeneous mass of fluid yolk in mature eggs. PMID- 15759287 TI - 3(10)-Helix adjoining alpha-helix and beta-strand: sequence and structural features and their conservation. AB - Does the amino acid use at the terminal positions of an alpha-helix become altered depending on the context-more specifically, when there is an adjoining 3(10)-helix, and can a single helical cylinder encompass the resultant composite helix? An analysis of 138 and 107 cases of 3(10)-alpha and alpha-3(10) composite helices, respectively, found in known protein structures indicate that the secondary structural element occurring first imposes its characteristics on the sequence of the structural element coming next. Thus, when preceded by a 3(10) helix, the preference of proline to occur at the N1 position of an alpha-helix is shifted to the N2 position, a typical characteristic of the C-terminal capping of the 3(10)-helix. When an alpha- or a 3(10)-helix leads into a helix of the other type, there is a bend at the junction, especially for the 3(10)-alpha composite, with the two junction residues facing inward and buried within the structure. Thus a single helical cylinder may not properly represent a composite helix, the bend providing a means for the tertiary structure to assume a globular shape, very much akin to what a proline-induced kink does to an alpha-helix. The tertiary structural context in which beta-3(10) and 3(10)-beta composites occurs can be different, causing the angle between the secondary structural elements in the two cases to be different. Composites of 3(10)-helices and beta-strands are much more conserved among members in families of homologous structures than those between two types of helices; in many of the former instances, the 3(10)-helix constitutes the loops in beta-hairpin or beta-beta-corner motifs. The overall fold of the chain may be more conserved than the actual identify of the secondary structure elements in a composite. PMID- 15759288 TI - Applying the stochastic difference equation to DNA conformational transitions: a study of B-Z and B-A DNA transitions. AB - Despite the existence of numerous models to account for the B-Z DNA transition, experimenters have not yet arrived at a conclusive answer to the structural and dynamical features of the B-Z transition. By applying the stochastic difference equation to simulate the B-Z DNA transition, we have shown that the stretched intermediate model of the B-Z transition is more probable than other B-Z transition models such as the Harvey model. This is accomplished by comparing potential energy profiles of various B-Z DNA transition models and calculating relative probabilities based on the stochastic difference equation with respect to length (SDEL) formalism. The results garnered in this article allow for new approaches in determining the structural transition of B-DNA to Z-DNA experimentally. We have also simulated the B-A DNA transition using the stochastic difference equation. Unlike the B-Z DNA transition, the mechanism for the B-A DNA transition is well established. The variation in the pseudorotation angle during the transition is in good agreement with experimental results. Qualitative features of the simulated B-A transition also agree well with experimental data. The SDEL approach is thus a suitable numerical technique to compute long-time molecular dynamics trajectory for DNA molecules. PMID- 15759289 TI - Enthalpy relaxation of bovine serum albumin and implications for its storage in the glassy state. AB - Two endothermic peaks could be observed for five commercial samples of bovine serum albumin (BSA). The smaller peak observed by differential scanning calorimetry (DSC) corresponded to enthalpy relaxation. This peak was followed on storage of BSA, in its glassy state, after it had been heated above its denaturation temperature. Enthalpy and peak temperature increased with duration of storage. On storage for one week at 60 degrees C, a sample at 8.3% moisture showed a peak at 100 degrees C with an energy value of approximately 2 J per g protein. BSA samples were heated within the DSC sufficiently to eliminate the lower enthalpy peak but without altering the denaturation enthotherm. The amount of physical aging shown by these BSA samples was similar to that of the heat denatured samples. It was concluded that the heating endotherms of dry BSA reflect both the storage and thermal history of the sample. Possible implications of the enthalpy relaxation of BSA on the behavior of this important protein are considered. PMID- 15759290 TI - Vibrations-determined properties of green fluorescent protein. AB - The physicochemical characteristics of the green fluorescent protein (GFP), including the thermodynamic properties (entropy, enthalpy, Gibbs' free energy, heat capacity), normal mode vibrations, and atomic fluctuations, were investigated. The Gaussian 03 computational chemistry program was employed for normal mode analysis using the AMBER force field. The thermodynamic parameters and atomic fluctuations were then calculated from the vibrational eigenvalues (frequencies) and eigenvectors. The regions of highest rigidity were shown to be the beta-sheet barrel with the central alpha-helix, which bears the chromophore. The most flexible parts of the GFP molecule were the outlying loops that cover the top and bottom of the beta-barrel. This way, the balance between rigidity and flexibility is maintained, which is the optimal relationship for protein stability in terms of Gibbs' free energy. This dual-schemed structure satisfies the requirements for GFP function. In this sense, the structure of GFP resembles a nanoscale drum: a stiff cylinder with flexible vibrating end(s). PMID- 15759291 TI - A study paradigm allowing comparison of multiple high-resolution rCBV-maps for the examination of drug effects. AB - Owing to the neuro-vascular coupling, measurement of changes in regional cerebral blood flow and blood volume (rCBV) can be used as surrogates reflecting the effects of central nervous system active drugs on neural transmission. As most such drugs are administered orally or intramuscularly and, in many cases, beneficial effects due to drug treatment can be observed only after chronic administration for days or weeks, the evaluation of drug efficacy requires the development of acquisition and analysis tools that allow for comparison of imaging data sets obtained in multiple sessions and for multiple subjects. In the present study, high-resolution susceptibility contrast MR perfusion imaging using a super-paramagnetic contrast agent (CA) was applied to study the effect of a single oral administration of the acetylcholine-esterase inhibitor rivastigmine (Exelon) on rCBV in rats. rCBV maps were calculated from two T2-weighted three dimensional fast-spin-echo scans recorded before and after the injection of the CA, respectively. All MRI data sets were mapped to a reference data set obtained from a normal male Sprague-Dawley rat using an automated co-registration procedure prior to the analysis for drug effects. Rivastigmine was orally administered at doses of 2, 4 or 8 mg/kg 1 h prior to the rCBV measurement. Rivastigmine increased rCBV in several brain areas including cortex, caudate putamen and hippocampus. The observed effects were dose-dependent and the changes reached the order of 5-12% as compared with baseline levels. Vehicle-treated animals showed no significant alterations of blood volume, demonstrating the reproducibility and stability of rCBV measurements. PMID- 15759292 TI - (1)H NMR spectroscopy in the diagnosis of Pseudomonas aeruginosa-induced urinary tract infection. AB - The utility of (1)H NMR spectroscopy is suggested and demonstrated for the diagnosis of Pseudomonas aeruginosa in urinary tract infection (UTI). The specific property of P. aeruginosa of metabolizing nicotinic acid to 6 hydroxynicotinic acid (6-OHNA) is exploited. The quantity of 6-OHNA produced correlates well with the viable bacterial count. Other common bacteria causing UTI such as Escherichia coli, Klebsiella pneumonia, Enterobacter aerogenes, Acinetobacter baumanii, Proteus mirabilis, Citrobacter frundii, Enterococcus faecalis, Streptococcus gp B and Staphylococcus aureus do not metabolize nicotinic acid under similar conditions. The method provides a single-step documentation of P. aeruginosa qualitatively as well as quantitatively. The NMR method is demonstrated on urine samples from 30 patients with UTI caused by P. aeruginosa. PMID- 15759296 TI - Anatomical and functional brain imaging using high-resolution echo-planar spectroscopic imaging at 1.5 Tesla. AB - High-resolution echo-planar spectroscopic imaging (EPSI) of water resonance (i.e. without water suppression) is proposed for anatomic and functional imaging of the human brain at 1.5 T. Water spectra with a resolution of 2.6 Hz and a bandwidth of 333 Hz were obtained in small voxels (1.7 x 1.7 x 3 mm3) across a single slice. Although water spectra appeared Lorentzian in most of the voxels in the brain, non-Lorentzian broadening of the water resonance was observed in voxels containing blood vessels. In functional experiments with a motor task, robust activation in motor cortices was observed in high-resolution T2* maps generated from the EPSI data. Shift of the water resonance frequency occurred during neuronal activation in motor cortices. The activation areas appeared to be more localized after excluding the voxels in which the lineshape of the water resonance had elevated T2* and became more non-Lorentzian during the motor task. These preliminary results suggest that high-resolution EPSI is a promising tool to study susceptibility-related effects, such as BOLD contrast, for improved anatomical and functional imaging of the brain. PMID- 15759298 TI - Analysis of nucleic acid constituents by on-line capillary electrophoresis-mass spectrometry. AB - This review is focused on the capillary electrophoresis-mass spectrometric (CE MS) analysis of nucleic acid constituents in the broadest sense, going from nucleotides and adducted nucleotides over nucleoside analogues to oligonucleotides. These nucleic acid constituents play an important role in a variety of biochemical processes. Hence, their isolation, identification, and quantification will undoubtedly help reveal the process of life and disease mechanisms, such as carcinogenesis, and can also be useful for antitumor and antiviral drug research to provide valuable information about mechanism of action, pharmacokinetics, pharmacodynamics, toxicity, therapeutic drug level monitoring, and quality control related to this substance class. Fundamental investigations into their structure, the search for modifications, the occurrence and biochemical impact of structural variation amongst others, are therefore of great value. In view of the related bioanalytical procedures, the coupling of CE to MS has emerged as a powerful tool for the analysis of the complex mixtures of nucleic acid constituents: CE confers rapid analysis and efficient resolution, while MS provides high selectivity and sensitivity with structural characterization of minute amounts of compound. After an introduction about the biochemical and analytical perspectives on the nucleic acid constituents, the different modes of CE used in this field of research as well as the relevant CE MS interfaces and the difficulties associated with quantitative CE-MS are briefly discussed. A large section is finally devoted to field-oriented applications. PMID- 15759299 TI - Chiral capillary electrophoresis-mass spectrometry of amino acids in foods. AB - In this work, the development of a new chiral capillary electrophoresis-mass spectrometry (CE-MS) method to separate D- and L-amino acids is shown. On-line coupling between CE and MS is established through an electrospray-coaxial sheath flow interface. Enantiomer separation is achieved by using a cheap, nonvolatile, chiral selector as beta-cyclodextrin in the background electrolyte (BGE) together with a physically coated capillary that is aimed to prevent contamination of the electrospray. The capillary coating is simple and easy to obtain as it only requires flushing of the capillary with a polymer aqueous solution for 3 min. Optimization of CE parameters (pH of BGE, type and concentration of chiral selector, and capillary inner diameter) and electrospray-MS parameters (nature and flow rate of the sheath liquid, nebulizer pressure) is carried out. Two different derivatization protocols of amino acids using dansyl chloride (DNS) and fluorescein isothiocyanate (FITC) are compared in terms of MS sensitivity and chiral resolution. Under optimum CE-MS conditions it is observed that the MS sensitivity obtained for FITC- and DNS-amino acids is similar (with limit of detection (LOD) in the microM range, corresponding to amounts injected in the fmol range) while chiral resolution is better for FITC-amino acids. The optimized method is demonstrated to provide the simultaneous analysis of 15 selected amino acids (i.e., FITC-D/L-Asp, -Glu, -Ser, -Asn, -Ala, -Pro, -Arg, and FITC-gamma aminobutyric acid (GABA) in a single chiral CE-MS run, corresponding to the main amino acids that can be found in orange. Moreover, as a result of the high resolution achieved, it is possible to detect down to 2% of D-Asp in the presence of 98% of L-Asp. The good possibilities of chiral CE-MS in food analysis are corroborated through the detection of the main amino acids in a commercial orange juice (i.e., FITC-L-Asp, -Glu, -Ser, -Asn, -Pro, -Arg, and the nonchiral FITC GABA) as well as the determination of the fraudulent addition of synthetic amino acids (containing D- and L-forms) to a fresh orange juice. PMID- 15759297 TI - Comparisons of brain metabolites observed by HRMAS 1H NMR of intact tissue and solution 1H NMR of tissue extracts in SIV-infected macaques. AB - The objective of this study was to compare ex vivo proton high-resolution magic angle spinning magnetic resonance spectra of intact tissue with those spectra obtained by solution (1)H NMR of brain extracts of the same sample. Sixteen brain tissue samples from simian immunodeficiency virus-infected rhesus macaques from both frontal cortex and putamen were evaluated by comparing brain metabolite quantities of N-acetylaspartate (NAA), choline-containing compounds (Cho), myo inositol (MI), creatine (Cr), lactate (Lac), glutamate (Glu) and acetate (Ace). The ratios of the individual NMR peak areas of all metabolites relative to the creatine peak area were calculated. Linear regression analysis revealed significant correlations between measurements using the two methods. The strength of the correlations varied depending on the metabolite studied. We found highly significant correlations for NAA/Cr (r2 = 0.77; p < 0.0001), NAA + Ace/Cr (r2 = 0.73; p < 0.0001) and MI/Cr (r2 = 0.75; p < 0.0001). We observed somewhat less strong correlations for Glu/Cr (r2 = 0.54; p < 0.002) and Lac/Cr (r2 = 0.54; p < 0.002). There was a substantially weaker correlation for Cho/Cr (r2 = 0.32; p = 0.02). When plotting the metabolite ratios obtained by 1H HRMAS NMR of the intact tissue sample on the ordinate vs 1H NMR of the tissue extract on the abscissa, most metabolites exhibited a slope close to unity, and a positive intercept probably due to macromolecular contributions to the MAS spectra. The slope for Cho/Cr was substantially less than unity. Generally, samples from the frontal cortex showed a better correlation between intact and extracted tissue samples than putamen. This is most prominent in the cases of NAA/Cr and Cho/Cr. We conclude that both methods provide substantially the same information for most major brain metabolites, with the exception of the Cho resonance. PMID- 15759300 TI - A soft on-column metal coating procedure for robust sheathless electrospray emitters used in capillary electrophoresis-mass spectrometry. AB - An on-column metal coating procedure was developed for sheathless electrospray emitters, based on Justus von Liebig's electroless silver mirror reaction followed by electrochemical deposition of gold onto the silver layer. The coating procedure is straightforward, mild, inexpensive, and can be performed with standard laboratory equipment. A long-term (600 h) stability investigation of the conductive coating was carried out by continuous electrospray in the positive electrospray mode, and no degradation in performance was found. The simplicity of the coating procedure and the robustness of the spray tips makes the spray tips highly suitable to couple delicate wall-coated or monolithic capillary columns to mass spectrometry. Peptide mixtures were separated by capillary electrophoresis and injected into either a Hadamard-transform time-of-flight mass analyzer or a commercial quadrupole mass analyzer using the described sheathless electrospray emitters. The performance was judged to be excellent. PMID- 15759301 TI - Glycoform characterization of intact erythropoietin by capillary electrophoresis electrospray-time of flight-mass spectrometry. AB - Glycosylated proteins often show a large variation in their glycosylation pattern, complicating their structural characterization. In this paper, we present a method for the accurate mass determination of intact isomeric glycoproteins based on capillary electrophoresis-electrospray-time of flight-mass spectrometry. Human recombinant erythropoietin has been chosen as a showcase. The approach enables the on-line removal of nonglycosylated proteins, salts, and neutral and negatively charged species. More important, different glycosylation forms are separated both on the base of differences in the number of negatively charged sialic acid residues and the size of the glycans. Thus, 44 glycoforms and in total about 135 isoforms of recombinant human erythropoietin, taking also acetylation into account, could be distinguished for the reference material from the European Pharmacopeia. Distinct glycosylation differences for samples from different suppliers are clearly observed. Based on the accurate mass an overall composition of each single isoform is proposed, perfectly in agreement with data on glycan and glycopeptide analysis. This method is an ideal complement to the established techniques for glycopeptide and glycan analysis, not differentiating branching or linkage isoforms, but leading to an overall composition of the glycoprotein. The presented strategy is expected to improve significantly the ability to characterize and quantify isomeric glycoforms for a large variety of glycoproteins. PMID- 15759302 TI - Speciation of arsenic compounds in fish and oyster tissues by capillary electrophoresis-inductively coupled plasma-mass spectrometry. AB - A capillary electrophoresis-inductively coupled plasma-mass spectrometric (CE-ICP MS) method for the speciation of six arsenic compounds, namely arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid, dimethylarsinic acid, arsenobetaine and arsenocholine is described. The separation has been achieved on a 70 cm length x 75 microm ID fused-silica capillary. The electrophoretic buffer used was 15 mM Tris (pH 9.0) containing 15 mM sodium dodecyl sulfate (SDS), while the applied voltage was set at +22 kV. The arsenic species in biological tissues were extracted into 80% v/v methanol-water mixture, put in a closed centrifuge tube and kept in a water bath, using microwaves at 80 degrees C for 3 min. The extraction efficiencies of individual arsenic species added to the sample at 0.5 microg As/g level were between 96% and 107%, except for As(III), for which it was 89% and 77% for oyster and fish samples, respectively. The detection limits of the species studied were in the range 0.3-0.5 ng As/mL. The procedure has been applied for the speciation analysis of two reference materials, namely dogfish muscle tissue (NRCC DORM-2) and oyster tissue (NIST SRM 1566a), and two real world samples. PMID- 15759303 TI - Protein profiling by capillary isoelectric focusing, reversed-phase liquid chromatography, and mass spectrometry. AB - An automated system for intact protein analysis is described that combines capillary isoelectric focusing (CIEF), reversed-phase liquid chromatography (RPLC), and electrospray ionization-mass spectrometry (ESI-MS). Performance is demonstrated with a complex yeast enzyme concentrate. CIEF is performed with a microdialysis membrane-based cathodic cell that permits pI fractions to be sampled and stored for subsequent LC-MS analysis. A total of 50 microg protein is loaded onto the capillary. Ten fractions are stored which span the pI range 3-10. Each fraction is subsequently cleaned on a reversed-phase trap column and then characterized by LC-MS. MaxEnt1 is used to deconvolute the raw mass spectra to obtain the molecular weight (MW) of intact proteins/peptides in the sample. A two dimensional display of pI vs. MW is illustrated for the 500 most prevalent species as identified by MaxEnt1. PMID- 15759304 TI - Capillary electrophoresis and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry for analysis of the novel erythropoiesis-stimulating protein (NESP). AB - NESP (novel erythropoiesis-stimulating protein) is a recently approved hyperglycosylated analogue of human erythropoietin (EPO) with a long-lasting effect. In this work, the capillary electrophoresis (CE) methodology proposed by the European Pharmacopoeia for the separation of EPO glycoforms has been modified for the separation of NESP glycoforms. Optimization of pH of the separation electrolyte has been fundamental in order to achieve baseline resolution of seven peaks corresponding to NESP glycoforms. Intact NESP has also been characterized by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). An accurate approximation to an average molecular mass of the NESP molecule has been obtained, taking into account the strong influence of laser intensity upon the MALDI-TOF mass spectra found. PMID- 15759305 TI - Quantitative analysis of six pesticides in fruits by capillary electrophoresis electrospray-mass spectrometry. AB - A method to identify and quantify six pesticide residues - dinoseb, pirimicarb, procymidone, pyrifenox, pyrimethanil, and thiabendazole - in peaches and nectarines using capillary electrophoresis-electrospray ionization-quadrupole ion trap-tandem mass spectrometry (CE-ESI-MS/MS) is described. Separation was carried out using a buffer of 0.3 M ammonium acetate at pH 4 with 10% methanol. Pesticide residues present in peach and nectarine samples were preconcentrated by solid phase extraction using C(18), eluted with CH(2)Cl(2), concentrated to dryness, and redissolved in buffer to obtain lower detection limits. The recoveries of the analytes ranged from 58 to 99% and the relative standard deviations were 9 to 19%. Under optimized CE-MS/MS conditions the minimum detectable levels for the six pesticides in spiked peach samples were between 0.01 mg/kg for pirimicarb and 0.05 mg/kg for procymidone with pressure injection of 50 mbar for 5 s (5 nL) at a signal-to-noise ratio of 3, which constitutes a severalfold increase in sensitivity compared to CE-MS, using a single quadrupole, and to conventional CE UV. The potential of the method was demonstrated by analyzing different samples taken from regional agricultural cooperatives. The pesticides most often detected were thiabendazole and procymidone. PMID- 15759306 TI - Anthocyanins increase low-density lipoprotein and plasma cholesterol and do not reduce atherosclerosis in Watanabe Heritable Hyperlipidemic rabbits. AB - Anthocyanin-rich beverages have shown beneficial effects on coronary heart disease in epidemiological and intervention studies. In the present study, we investigated the effect of black currant anthocyanins on atherosclerosis. Watanabe Heritable Hyperlipidemic rabbits (n = 61) were fed either a purified anthocyanin fraction from black currants, a black currant juice, probucol or control diet for 16 weeks. Purified anthocyanins significantly increased plasma cholesterol and low-density lipoprotein (LDL) cholesterol. Intake of black currant juice had no effect on total plasma cholesterol, but lowered very-low density lipoprotein (VLDL) cholesterol significantly. There were no significant effects of either purified anthocyanins or black currant juice on aortic cholesterol or development of atherosclerosis after 16 weeks. Probucol had no effect on plasma cholesterol but significantly lowered VLDL-cholesterol and decreased aortic cholesterol accumulation. The erythrocyte antioxidant enzyme glutathione peroxidase was significantly increased by purified anthocyanins and superoxide dismutase was increased by both anthocyanin-containing treatments. Other markers of plasma antioxidant capacity, antioxidant enzymes, protein and lipid oxidation were not affected by any of the anthocyanin treatments. Adverse effects of purified anthocyanins were observed on plasma- and LDL-cholesterol. These effects were not observed with black currant juice, suggesting that black currants may contain components reducing the adverse effects of anthocyanins. PMID- 15759307 TI - Analysis of melamine resins by capillary zone electrophoresis with electrospray ionization-mass spectrometric detection. AB - A method for the determination of the major components of (methoxymethyl)melamine resins, with quantitative analysis of unreacted melamine by capillary zone electrophoresis (CZE) using electrospray ionization-mass spectrometry (ESI-MS) is presented. Using a low background electrolyte (BGE) pH, components are separated according to their charge/ionic radius ratio with a distinctly different separation selectivity compared to the HPLC methods commonly employed in melamine resin analysis. The use of a time-of-flight mass spectrometer (TOF-MS) was concluded to be necessary, as the complex samples studied required maximum sensitivity and resolution, which is clearly superior for TOF-MS detectors over their quadrupole counterparts. A standard curve of free melamine was determined with an R(2) = 0.999 over a concentration range of an order of magnitude. This method offers the unique separation selectivity of CZE as well as a quicker analysis time, especially for dimers compared to the HPLC methods used to date. PMID- 15759308 TI - Mass spectral analysis of synthones of nerve agents for verification of the Chemical Weapons Convention. AB - This communication describes the synthesis and gas chromatography/mass spectrometric (GC/MS) analysis of N,N-dialkylphosphoramidic dihalides and alkylphosphonic difluorides, which are synthones of nerve agents. The study was undertaken with a view to developing a spectral database of these compounds for verification purposes of the Chemical Weapons Convention (CWC). The modified synthetic approach reported here has advantages over traditional syntheses in terms of time and yield. GC/MS analysis of these synthones yielded electron ionization (EI) mass spectra and, based on these spectra, generalized fragmentation routes are proposed that rationalize most of the characteristic ions. PMID- 15759309 TI - Inhibitors of the epidermal growth factor receptor in apple juice extract. AB - The polyphenol-rich extract of a consumer-relevant apple juice blend was found to potently inhibit the growth of the human colon cancer cell line HT29 in vitro. The epidermal growth factor receptor (EGFR) and its subsequent signaling cascade play an important role in the regulation of cell proliferation in HT29 cells. The protein tyrosine kinase activity of an EGFR preparation was effectively inhibited by the polyphenol-rich apple juice extract. Treatment of intact cells with this extract resulted in the suppression of the subsequent mitogen-activated protein kinase cascade. Amongst the so far identified apple juice constituents, the proanthocyanidins B1 and B2 as well as quercetin-3-glc (isoquercitrin) and quercetin-3-gal (hyperoside) were found to possess substantial EGFR-inhibitory properties. However, as to be expected from the final concentration of these potential EGFR inhibitors in the original polyphenol-rich extract, a synthetic mixture of the apple juice constituents identified and available so far, including both proanthocyanidins and the quercetin glycosides, showed only marginal inhibitory effects on the EGFR. These results permit the assumption that yet unknown constituents contribute substantially to the potent EGFR-inhibitory properties of polyphenol-rich apple juice extract. In summary, the polyphenol composition of apple juice possesses promising growth-inhibitory properties, affecting proliferation-associated signaling cascades in colon tumor cells. PMID- 15759310 TI - Effective depletion of albumin using a new peptide-based affinity medium. AB - Blood plasma and serum are very useful samples for the detection, identification and quantitation of proteins associated with both health and disease. However, analysis of plasma and serum is a challenge because traces of interesting polypeptides and proteins can be dominated by the very high concentration of albumin present. Albumin may be depleted by adsorption to immunoaffinity columns or to columns containing dyes such as Cibacron Blue, or by ultrafiltration, but these methods are far from ideal. We describe a new peptide-based affinity medium which is effective for removing albumin and is very specific. The albumin-binding capacity is at least 14 mg per mL of gel. The material may be reused hundreds of times after a simple regeneration step involving NaOH, with full retention of specificity and capacity. The material was tested with human and monkey plasma and serum and rat serum, and has been used to deplete litre volumes of human plasma. The development of other peptide-based affinity media to deplete abundant proteins is briefly discussed. PMID- 15759311 TI - Electrospray ionization mass spectrometric study of purine base-cisplatin complexes. AB - By mixing cisplatin (cis-diamminedichloroplatinum(II)) with purine base the following ions have been obtained under electrospray ionization conditions: [A+Pt(NH3)2 Cl]+, [A+PtNH3Cl]+, [G+Pt(NH3)2 Cl]+ and [G+PtNH3)Cl]+. Their collision-induced dissociation led to the loss of NH3 and HCl and formation of the protonated base. The last process is strongly favoured for adenine over guanine. It confirms that, analogously as for DNA, formation of the guanine cisplatin complex is favoured over that of the adenine complex and, as a consequence, it suggests that the mass spectrometric study of nucleic base complexes with platinum may provide some information on the interactions of DNA with other platinum drugs. The loss of NH3 accompanied by that of CO from the guanine ring has experimentally confirmed the presence of a strong hydrogen bond between the NH3 molecule and the O=C6 moiety of guanine found by theoretical calculations. PMID- 15759312 TI - Free and total para-aminobenzoic acid analysis in plants with high-performance liquid chromatography/tandem mass spectrometry. AB - para-Aminobenzoic acid (PABA), a precursor in the synthesis of folates in plants, is determined by liquid chromatography/tandem mass spectrometry (LC/MS/MS). In plants PABA can be converted into its beta-D-glucopyranosyl ester (PABA-Glc) and can also exist in its free form. In this work, we developed and validated a quantitative method to study free and total PABA in plants. The total PABA (free PABA plus PABA-Glc) can be evaluated after acid hydrolysis at 80 degrees C for 2 hours. The plant material is homogenized and the PABA content is quantified using the standard addition procedure. The validated method is selective, sensitive, simple, accurate, has a recovery between 99.6 to 102.5%, is reproducible (RSD between 1.4 and 4.4%), and is linear between 2.5 and 1538 ng/mL. Free and total PABA determinations in five vegetables showed that different plant species had different amounts of free and total PABA, and that the ratios of total versus free PABA were also variable. This new method could be valuable for studies of folate synthesis in plants. PMID- 15759313 TI - Tryptic transpeptidation products observed in proteome analysis by liquid chromatography-tandem mass spectrometry. AB - Commonly, prior to mass spectrometry based analysis of proteins or protein mixtures, the proteins are subjected to specific enzymatic proteolysis. For this purpose trypsin is most frequently used. However, the process of proteolysis is not unflawed. For example, some side activities of trypsin are known and have already been described in the literature (e.g., chymotryptic activity). Here, we describe the occurrence of transpeptidated peptides during standard proteome analysis using two-dimensional polyacrylamide gel electrophoresis followed by mass spectrometric protein identification. Different types of transpeptidated peptides have been detected. The most frequently observed transpeptidation reaction is N-terminal addition of arginine or lysine to peptides. Furthermore, addition of two amino acids to the N-terminus of a peptide has also been detected. Another transpeptidation that we observed, is combination of two peptides, which were originally located in different regions of the analyzed protein. Currently, the full amount of peptides generated by transpeptidation is not clear. However, it should be recognized that protein information is presently lost as these effects are not detectable with available database search software. PMID- 15759314 TI - Global protein expression analysis in apicomplexan parasites: current status. AB - Members of the phylum Apicomplexa are important protozoan parasites that cause some of the most serious, and in some cases, deadly diseases in humans and animals. They include species from the genus Plasmodium, Toxoplasma, Eimeria, Neospora, Cryptosporidium, Babesia and Theileria. The medical, veterinary and economic impact of these pathogens on a global scale is enormous. Although chemo- and immuno-prophylactic strategies are available to control some of these parasites, they are inadequate. Currently, there is an urgent need to design new vaccines or chemotherapeutics for apicomplexan diseases. High-throughput global protein expression analyses using gel or non-gel based protein separation technologies coupled with mass spectrometry and bioinformatics provide a means to identify new drug and vaccine targets in these pathogens. Protein identification based proteomic projects in apicomplexan parasites is currently underway, with the most significant progress made in the malaria parasite, Plasmodium falciparum. More recently, preliminary two-dimensional gel electrophoresis maps of Toxoplasma gondii and Neospora caninum tachyzoites and Eimeria tenella sporozoites, have been produced, as well as for micronemes in E. tenella. In this review, the status of proteomics in the analysis of global protein expression in apicomplexan parasites will be compared and the challenges associated with these investigations discussed. PMID- 15759315 TI - Biochemical and immunological studies of nucleocapsid proteins of severe acute respiratory syndrome and 229E human coronaviruses. AB - Severe acute respiratory syndrome (SARS) is a serious health threat and its early diagnosis is important for infection control and potential treatment of the disease. Diagnostic tools require rapid and accurate methods, of which a capture ELISA method may be useful. Toward this goal, we have prepared and characterized soluble full-length nucleocapsid proteins (N protein) from SARS and 229E human coronaviruses. N proteins form oligomers, mostly as dimers at low concentration. These two N proteins degrade rapidly upon storage and the major degraded N protein is the C-terminal fragment of amino acid (aa) 169-422. Taken together with other data, we suggest that N protein is a two-domain protein, with the N terminal aa 50-150 as the RNA-binding domain and the C-terminal aa 169-422 as the dimerization domain. Polyclonal antibodies against the SARS N protein have been produced and the strong binding sites of the anti-nucleocapsid protein (NP) antibodies produced were mapped to aa 1-20, aa 150-170 and aa 390-410. These sites are generally consistent with those mapped by sera obtained from SARS patients. The SARS anti-NP antibody was able to clearly detect SARS virus grown in Vero E6 cells and did not cross-react with the NP from the human coronavirus 229E. We have predicted several antigenic sites (15-20 amino acids) of S, M and N proteins and produced antibodies against those peptides, some of which could be recognized by sera obtained from SARS patients. Antibodies against the NP peptides could detect the cognate N protein clearly. Further refinement of these antibodies, particularly large-scale production of monoclonal antibodies, could lead to the development of useful diagnostic kits for diseases associated with SARS and other human coronaviruses. PMID- 15759316 TI - Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma: Identification of potential tumor markers. AB - Hepatocellular carcinoma (HCC) is a malignancy of both underdeveloped and developing countries. Proteomes of ten pairs of clinical hepatitis B virus associated HCC tissue samples were obtained by high resolution two-dimensional gel electrophoresis. Comprehensive analyses of proteins associated with B-type HCC were focused on total differentially expressed proteins (> or = two-fold increase or decrease, Student's t-test, p < 0.05) from one pair of samples. Protein identification was done by peptide mass fingerprinting with matrix assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-tandem mass spectrometry. Comparative analyses of proteins associated with B-type HCC included repeat statistics in ten cases. A total of 100 protein spots, corresponding to 80 different gene products, were identified. Proteins whose expression levels were different by more than 2-fold in at least 50% of the cases (five of ten cases) were further analyzed and 45 proteins were selected out as candidates for HCC-associated proteins. Western blotting further validated up-regulated expressions of two candidate proteins in tumor tissues: proliferating cell antigen and stathmin 1. This comprehensive and comparative analyses of proteins associated with B-type HCC could provide useful molecular markers for diagnostics and prognostics and for therapeutic targets. The physiological significance of the differential expressions for several candidate proteins are discussed. PMID- 15759317 TI - Identification of the heterogeneous nuclear ribonucleoprotein A2/B1 as the antigen for the gastrointestinal cancer specific monoclonal antibody MG7. AB - MG7 is an early gastrointestinal cancer specific monoclonal antibody. It can detect gastric cancer with high sensitivity and specificity. However, the target antigen for MG7 has not been identified. Western blot analysis revealed that the MG7 antibody reproducibly recognized two approximately 35 kDa proteins in the total cell lysates of human gastric carcinoma cell lines KATO III and MKN-45. Using a proteomic approach, we identified these MG7 immunoreactive proteins as the human heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1). Western blot analysis of nuclear and cytosolic fraction of KATO III cells using either MG7 or hnRNP A2/B1 antibodies confirmed that the target antigen is located exclusively in the nucleus. With the use of archival samples, we also found that the level of hnRNP A2/B1 protein was increased in gastric cancer tissues (4 out of 5 patients), when compared to their corresponding matching normal stomach tissue. PMID- 15759318 TI - Differential protein expression in human gliomas and molecular insights. AB - Gliomas are the most common of the primary intracranial tumors with astrocytomas constituting about 40%. Using clinically and histologically assessed astrocytomas, we have studied their protein profiles using a two-dimensional gel electrophoresis-mass spectrometry approach and identified differentially expressed proteins which may be useful molecular indicators to understand these tumors. Examination of the protein profiles of 27 astrocytoma samples of different grades revealed 72 distinct, differentially expressed proteins belonging to various functional groups such as cytoskeleton and intermediate filament proteins, heat shock proteins (HSPs), enzymes and regulatory proteins. Based on the consistency of their differential expression, 29 distinct proteins could be short-listed and may have a role in the pathology of astrocytomas. Some were found to be differentially expressed in both Grade III and IV astrocytomas while others were associated with a particular grade. A notable observation was underexpression of Prohibitin, a potential tumor suppressor protein, Rho-GDP dissociation inhibitor, Rho-GDI, a regulator of Rho GTPases and HSPs as well as destabilization of glial fibrillary acidic protein, GFAP, major protein of the glial filaments, in Grade III malignant tumors. We attempt to explain glioma malignancy and progression in terms of their combined role. PMID- 15759322 TI - The development of an algorithm for the mass spectral interpretation of phosphoproteins. AB - Extended from the peptide mapping method (Fenyo, D., Curr. Opin. Biotechnol. 2000, 11, 391-395) the proposed algorithm takes the mass information of a precursor ion to reconstruct all possible phosphorylated peptide sequences. The mass spectra of product ions from the corresponding precursor ion is then used and compared to the reconstructed sequences to deduce the most probable phosphoprotein. The proposed algorithm also predicts all possible combinations of phosphopeptides, which may serve as a clue for designing proper phosphorylation experiments to validate the existence of these peptides and the corresponding proteins. PMID- 15759324 TI - Challenges to WHI and MWS continue. PMID- 15759323 TI - Termination of the oestrogen-alone arm of the Women's Health Initiative. PMID- 15759325 TI - CT diagnosis of Meckel diverticulum in a paracolic internal hernia. AB - The autopsy incidence of internal hernia has been reported to be between 0.2% and 0.9%, and these hernias are usually diagnosed on imaging due to their complications or at surgery. Meckel diverticulum is the most common congenital anomaly of the gastrointestinal tract, occurring in 1% to 3% of the population according to autopsy studies. The condition also is usually diagnosed at surgery, by barium studies or scintigraphy, or on cross-sectional imaging due to complications. We present an unusual case of a large Meckel diverticulum in a right paracolic hernia diagnosed on multidetector computed tomography. This diagnosis was made after attacks of subacute intestinal obstruction with the aid of multiplanar reconstructions. This case emphasizes the role of multidetector computed tomography and postprocessing techniques such as multiplanar reconstruction in the diagnosis of bowel pathology. PMID- 15759326 TI - Primary and secondary malignancies of the penis: ultrasound features. AB - Cancer of the penis is a rare neoplasm in developed countries but worldwide represents a significant health problem. In this study, the ultrasonographic features of primary and secondary malignant lesions of the penis are described. Squamous cell carcinoma usually presents as a hypoechoic lesion with heterogeneous appearance. Invasions of the corpora cavernosa and the corpus spongiosum are appreciable. B-cell lymphoma presents as well-vascularized mass, a plaque, or ulcers in the penile skin. Penile metastases results from hematogenous or lymphatic spreading of distant tumors or, more frequently, as penile infiltration by tumors from adjacent organs. Diffuse corporeal or nodular involvement can result. PMID- 15759327 TI - Fine print TRIPS up multinational and Indian companies. PMID- 15759328 TI - Facile degradative lactonization of Gln-Arg and Gln-Phe hydroxyethylene dipeptide derivatives. AB - We have found that hydroxyrthylene (HE) dipeptide analogs of Gln-Arg and Gln-Phe are usually susceptible to acid catalyzed lactonization. The synthesis of substrate-based transition state analog inhibitors of botulinum neurotoxin metalloprotease inhibitors that contain the Gln-Arg or the Gln-Phe HE units is complicated by this facile degradative lactonization. PMID- 15759329 TI - Impact of 2005 Medicare physician fee schedule on group practices. PMID- 15759330 TI - Position paper on safe staffing. PMID- 15759331 TI - Festschrift on the occasion of the 70th birthday of Allan S. Hoffman. Gels, genes, grafts and giants: Part 1. Symposium in Maui, Hawaii, USA, December 2002. PMID- 15759332 TI - NIH Conference on Frontiers of Rehabilitation Medicine: Osteogenesis Imperfecta. September 24-25, 1992, Bethesda, Maryland, USA. PMID- 15759333 TI - First self. PMID- 15759334 TI - Schistosoma mansoni in family 5 years after safari. PMID- 15759335 TI - Community associated methicillin-resistant Staphylococcus aureus, Singapore. PMID- 15759336 TI - Proceedings of the Mitochondria and Neuroprotection Symposium, in memory of Albert L. Lehninger. Fort Lauderdale, Florida, USA, April 16-19, 2004. PMID- 15759337 TI - Mumps virus-associated hemophagocytic syndrome. PMID- 15759338 TI - Imported cutaneous diphtheria, Germany, 1997-2003. PMID- 15759339 TI - Antimicrobial drug consumption in companion animals. PMID- 15759340 TI - Vibrio cholerae SXT element, Laos. PMID- 15759341 TI - Modeling the impact of pandemic influenza on Pacific Islands. PMID- 15759342 TI - Mycotic brain abscess caused by opportunistic reptile pathogen. PMID- 15759343 TI - Tuberculosis in undocumented migrants, Geneva. PMID- 15759344 TI - Mycobacterium chelonae skin infection in kidney-pancreas recipient. PMID- 15759345 TI - Diagnostic analysis of two sister pairs with endometrial cancer. PMID- 15759346 TI - SARS control and psychological effects of quarantine, Toronto, Canada. PMID- 15759347 TI - Commentary: bipolar disorder is a potentially fatal disease. PMID- 15759348 TI - Emergency airway equipment and training. PMID- 15759349 TI - Staging of renal cell carcinoma: current concepts. PMID- 15759350 TI - Laparoscopic specimen extraction: morcellation. AB - OBJECTIVES: To review our experience with intact extraction and morecellation of nephrectomy specimen, and the advantages and disadvantages of morcellation indicated by current reports. PATIENTS AND METHODS: In a previous study, 56 consecutive patients undergoing radical and simple transperitoneal laparoscopic nephrectomy were prospectively evaluated. Morcellation specimens (33) were extracted at the umbilical or lateral port sites and intact specimens (23) through an infraumbilical incision. Data were obtained on pathology, narcotic requirements, hospital stay, complications, estimated blood loss, size of renal mass based on preoperative imaging, specimen weight and extraction incision length. RESULTS: The mean incision length was 1.2 cm in the morcellation group and 7.1 cm in the intact group (P< 0.001). There were no significant differences in pain or recovery between the groups. In two cases of tumor nephrectomy, microscopic invasion of the perinephric adipose tissue in the intact specimen group were up-staged from clinical T1 to pT3a disease; there was no change in patient treatment based on this information. CONCLUSIONS: With proper technique, morcellation is safe for extracting renal tumours. The specimen can be evaluated for histology but not for pathological staging, limiting its use with transitional cell carcinoma. Port-site seeding is rare, and does not appear to be more frequent than with open nephrectomy. Although morcellation is cosmetically more desirable, there was no significant advantage in operating time, pain or duration of hospital stay. The choice od extraction method depends on the surgeon's preference and patient choice. PMID- 15759351 TI - Lymphadenectomy for renal cell carcinoma. PMID- 15759352 TI - Needle-ablative nephron-sparing surgery. PMID- 15759353 TI - Laparoscopic live donor nephrectomy: current status. PMID- 15759354 TI - Broadening experience with the retrograde endoscopic management of upper urinary tract urothelial malignancies. PMID- 15759355 TI - Staff shortages hampering UK cancer care. PMID- 15759356 TI - Lights on for patients with skin cancer. PMID- 15759357 TI - High leukaemia cure rate in Down's syndrome explained. PMID- 15759358 TI - Crackdown on cancer-cell resistance. PMID- 15759359 TI - Net-based cancer atlas shows new cancer trends in India. PMID- 15759360 TI - Viral turn-on for Kaposi's sarcoma cytokines. PMID- 15759361 TI - Bevacizumab adds survival benefit in colorectal cancer. PMID- 15759362 TI - Letrozole better than tamoxifen in postmenopausal women. PMID- 15759363 TI - New agent overcomes resistance to imatinib. PMID- 15759364 TI - Should adjuvant chemotherapy become standard treatment in all patients with resected non-small-cell lung cancer? AB - Surgery remains the main curative treatment for patients with early-stage non small-cell lung cancer (NSCLC); however, because many patients probably have undetectable micrometastasis even at diagnosis, adjuvant treatment is usually needed. The results for radiotherapy have mostly been disappointing, and a strong emphasis has, therefore, been placed on chemotherapy as the preferred modality. Adjuvant chemotherapy, and in particular, platinum-based regimens, have been assessed in several studies, but the results have been conflicting. Most trials have included patients with a wide range of disease stages and have shown, at most, only moderate improvements in survival. Thus, although clearly indicated in some patients, whether adjuvant chemotherapy should be used in all patients with resected disease is highly controversial. In this debate, Thierry Le Chevalier and colleagues and Giorgio Scagliotti present opposing arguments for whether this approach should be considered standard. PMID- 15759366 TI - On a class of integrals of Legendre polynomials with complicated arguments--with applications in electrostatics and biomolecular modeling. AB - The exact analytical result for a class of integrals involving (associated) Legendre polynomials of complicated argument is presented. The method employed can in principle be generalized to integrals involving other special functions. This class of integrals also proves useful in the electrostatic problems in which dielectric spheres are involved, which is of importance in modeling the dynamics of biological macromolecules. In fact, with this solution, a more robust foundation is laid for the Generalized Born method in modeling the dynamics of biomolecules. PMID- 15759365 TI - Non-random fluctuations and multi-scale dynamics regulation of human activity. AB - We investigate if known extrinsic and intrinsic factors fully account for the complex features observed in recordings of human activity as measured from forearm motion in subjects undergoing their regular daily routine. We demonstrate that the apparently random forearm motion possesses dynamic patterns characterized by robust scale-invariant and nonlinear features. These patterns remain stable from one subject to another and are unaffected by changes in the average activity level that occur within individual subjects throughout the day and on different days of the week, since they persist during daily routine and when the same subjects undergo time-isolation laboratory experiments designed to account for the circadian phase and to control the known extrinsic factors. Further, by modeling the scheduled events imposed throughout the laboratory protocols, we demonstrate that they cannot account for the observed scaling patterns in activity fluctuations. We attribute these patterns to a previously unrecognized intrinsic nonlinear multi-scale control mechanism of human activity that is independent of known extrinsic factors such as random and scheduled events, as well as the known intrinsic factors which possess a single characteristic time scale such as circadian and ultradian rhythms. PMID- 15759367 TI - Survey of palliative medicine fellowship programs, 2003-2005. PMID- 15759368 TI - Study on the platelet factor and beta-thromboglobulin in the patients with ischemic heart disease. PMID- 15759369 TI - Anti-thyroxine antibodies in autoimmune thyroiditis--effect on radioimmunoassay and binding characteristics. PMID- 15759370 TI - Observation of the serum uric acid in essential hypertension. PMID- 15759371 TI - A clinical study of adult Japanese encephalitis in the Chonnam District, Korea, during summer of 1982--a difference between improved and expired cases. PMID- 15759372 TI - The changes of serum-ACE, plasma renin activity and aldosterone in the diabetics with hypertension. PMID- 15759373 TI - The prevalence of antithyroid autoantibodies in normal Korean population--age, sex distribution and its relation to thyroid function. PMID- 15759375 TI - The studies on the gastrin levels in the patients with renal failure. PMID- 15759374 TI - Profiles of serum bile acids in liver diseases. PMID- 15759376 TI - The clinical and thyroid function studies of lymphocytic thyroiditis with spontaneously resolving hyperthyroidism: comparison to subacute thyroiditis. PMID- 15759377 TI - Ferrokinetics in patients on CAPD: influence of CAPD on the anemia of uremia. PMID- 15759378 TI - A study on cerebral embolism in mitral stenosis. PMID- 15759379 TI - Chemical liver function tests and epidemiologic studies of HBsAg positive blood donors. PMID- 15759380 TI - A study of the renin-angiotensin system and the blood volume in the nephrotic syndrome. PMID- 15759381 TI - Studies on changes of beta-adrenergic receptors in polymorphonuclear cell and mononuclear cell with the changes of thyroid function. PMID- 15759382 TI - 2,3-diphosphoglyceric acid changes in uremia and during hemodialysis. PMID- 15759383 TI - Prognostic evaluation of endoscopic sclerotherapy on bleeding esophageal varices. PMID- 15759384 TI - Effect of beta-endorphin and cortisol on the PHA stimulated lymphoblastogenesis. PMID- 15759385 TI - Changes of serum angiotensin-converting enzyme activity during treatment of patients with Graves' disease. PMID- 15759386 TI - Serum and erythrocyte lipoperoxides and supproxide dismutase levels in normal persons and hypertensive patients. PMID- 15759387 TI - Postprandial 2-hr C-peptide concentration as a guide for insulin treatment in patient with NIDDM. PMID- 15759388 TI - Acute transverse myelitis complicated in Korean hermorrhagic fever: a case report and review of the literature. PMID- 15759390 TI - Allelopathy. A natural strategy for weed control. PMID- 15759389 TI - Herbicide-resistant weeds in Europe: the wider implications. AB - Herbicide-resistance occurs in 55 weed species in 21 European countries. 91% of cases are associated with just four herbicide mode of action groups: ACCase and ALS inhibitors, and triazine and urea/amide photosynthetic inhibitors. There are also a few cases of resistance to bypiridiliums, dinitroanilnes and synthetic auxins. Resistance to ALS inhibitors tends to be less prevalent in Europe than elsewhere, but is likely to increase. A small scale survey showed that Alopecurus myosuroides is considered to be the most important herbicide-resistant weed in Europe at present. Lolium spp., and to a lesser extent Papaver rhoeas and Avena spp., were also highlighted as being of major importance in many countries. One consequence of the ongoing EC review of pesticides may be a reduction in the range of modes of action available to European farmers. This may reduce the opportunities for rotating different modes of action as a method of reducing resistance risk. Greater dependence on high resistance risk herbicides, such as ACCase and ALS inhibitors, because of lack of alternative modes of action, is likely to increase the incidence of resistance in grass-weeds. PMID- 15759391 TI - Carabid larvae as predators of weed seeds: granivory in larvae of Amara eurynota (Coleoptera: Carabidae). AB - Up to date we do not have much information about predation on seeds by larvae of ground beetles. One of the reasons why such knowledge is important is that granivorous larvae contribute to predation of weed seeds. In this study, the food requirements of larvae of autumn breeding carabid species Amara eurynota (Panzer) were investigated in the laboratory and a hypothesis, that they are granivorous was tested. Insect diet (Tenebrio molitor larvae), three seed diets (seeds of Artemisia vulgaris, Tripleurospermum inodorum or Urtica dioica or a mixed diet (T. molitor + A. uulgaris) were used as food. For larvae of A. eurynota, seeds are essential for successful completion of development, because all those fed pure insect diet died before pupation. However, differences in suitability were observed between pure seed diets. Larvae fed seeds of A. vulgaris had the lowest mortality and fastest development of the seed diets. Those fed seeds of T. inodorum had also low mortality, but the development was prolonged in the third instar. In contrast, development of larvae reared on seeds of U. dioica was slowest of the tested diets and could not be completed, as all individuals died before pupation. When insects were included to seed diet of A. vulgaris (mixed diet), the duration of development shortened, but mortality remained the same when compared to seed diet of A. vulgaris. According to the results it was concluded that larvae of A. eurynota are granivorous. A mixed diet and seed diets of A. uulgaris and T. inodorum were suitable and insect diet and seeds of U. dioica were unsuitable diets in this experiment. PMID- 15759392 TI - Isolation of simazine-degrading bacterial consortia from olive fields of andalucia. PMID- 15759393 TI - Soil activity and persistence of sulcotrione and mesotrione. AB - Greenhouse bioassays were set up using a small pot test method to determine the intrinsic sensitivity of different plant species to sulcotrione and mesotrione applied in a sandy loam soil. Herbicides were applied over an appropriate concentration range. After a 2-3 week test period, foliage fresh weight was determined. Data were subjected to a non-lineair regression analysis. Using the regression equations, ED50-values (herbicide concentrations that cause 50 percent foliage fresh weight reduction) were calculated for each combination of crop species and herbicide. To determine which replacement crops might be grown in case of failure of a crop treated with one of these herbicides, field persistence experiments were conducted over the 1993-2003 period for sulcotrione and the 1998 2003 period for mesotrione at the Experimental Farm, Biocentre Agri-Vet, Ghent University at Melle. Herbicides were applied in spring (about mid-March) on a bare soil; untreated control strips were included. Replacement crops were sown or planted approximately five weeks after herbicide applications. Visual estimations of crop injury were recorded at several intervals from sowing and fresh matter yield of plant parts was determined. Based on these data, crops were ranked according to their degree of sensitivity to either sulcotrione or mesotrione. Maize is very tolerant to both herbicides, although in some years, temporary injury could be seen in the field experiments. Italian rye-grass and fibre flax are tolerant crops; in field experiments a slight, temporary injury could be noticed in some years. Winter wheat displayed a high degree of tolerance to mesotrione (in both experiment types): however this crop was less tolerant to sulcotrione especially in the bioassay experiment. Based on its ED50-value, black salsify is tolerant to sulcotrione but under field conditions, the selectivity of this herbicide is quite variable; tolerance to mesotrione is moderate. Turnip and witloof chicory are clearly sensitive to mesotrione and sulcotrione whereas sugar beet, red clover and lettuce are extremely sensitive to both herbicides in both experiment types. Bioassays and field experiments provide a detailed and complete information about soil activity and persistence of both herbicides. PMID- 15759394 TI - Effect of synthetic combinations and tobacco root extract and the mixture of them on broomrape seed germination. AB - Various types of broomrape (Orobanche cermua), such as parasitic weeds, live on the roots of many important crops. Different methods may be advised to eradicate this parasite. But the seed size is so tiny with high stability in the soil and having strong connection with the host, it is very difficult to eradicate this parasite and there is no useful and economic method has not been suggested in this regard yet. The seeds of all types of broomrape germinate in the soil only when they are exposed to the secretions of the host's roots, therefore, providing special combinations that cause the parasite's seeds germinate in the absence of the host, can be an effective way in controlling this parasite. The effect of the tobacco root's extract and the synthetic combinations as well as their mixtures with different concentrations on the germination of the seeds of Orobanche cermua Loefi were tested to introduce an effective combination that can be used in controlling this parasite. This experiment includes 28 treatments in three replications. The tobacco root's extract was derived from the tobacco roots planted in pots containing Perlit in different stages and were prepared in different concentrations. Synthetic combinations (including combinations 1, 2 and 3) were also prepared with concentration of 1, 2 and 3 parts per million (ppm), then these combinations were tested as treatments on Broomrape's seeds. At the end of experiment, it was concluded that all the combinations used in different concentrations in this experiment significantly differ from the control treatment in their effects in causing the Broomrape's seeds germinate. Among all the combinations examined, the following were the most effective ones in the process of the germination of the Broomrape's seeds: 1. combination 2 with the concentration of one part per million, 2. combination 2 with the concentration of two parts per million, and 3 tobacco root's extract in the third stage. PMID- 15759395 TI - Experimental and natural weed host-virus relations. AB - Weeds, as alternative hosts of plant viruses and nutrient plants of virus vectors play important role in virus ecology and epidemiology. The aim of our study was to discover new weed-virus relations. Therefore some weed species were mechanically inoculated with 28 viruses (strains or isolates) maintained in our glasshouse. Different weed species with and without visible symptoms were collected from agro-, water ecosystems and wastelands of Hungary between 1997 and 2003. Virus infections were evaluated by biotests, DAS ELISA serological methods, electronmicroscopy and immunosorbent electronmicroscopy (ISEM). Under glasshouse conditions Ambrosia artemisifolia was considered as a virophob species, showing resistance to all viruses listed above. A series of new artificial (Chenopodium album--SoMV (LH+SH)*, AMV (LH+SH); C. berlandieri--PVY(NTN) (LH), AMV (LH+SH), CMV (LH), SoMV (LH+SH), ObPV (LH+SH), ZYMV-10 (LH): C. ugandae--ObPV (LH), SoMV (L); C. glaucum--ObPV (LH), SoMV (L); Echinocystis lobata--PVX (L), ZYMV (LH+SH); Solanum nigrum--MYFV (LH+SH), PVY(N) (L), PVY(NTN) (LH+SH), SoMV (LH), TMV (SH), CMV (SH); S. dulcamara--CMV-U/246 (SH), PVY(NTN) (LH), SoMV-H (L), TMV-O (L); S. luteum--PVY(N) (SH), PVY(NTN) (LH+L), TMV(SH).) and natural (Asclepias syriaca- TMV, AMV, TSWV; Alisma plantago-aquatica--PVY, SoMV; Ambrosia artemisiifolia- CMV; Chenopodium album--CMV, PVS, PLRV; C. hybridum--CMV; Cirsium canum--CMV, PVM; Carex vulpina--CMV; Comium maculatum--PVY; Datura stramonium--PVA, PVX, PVS, PVM, CMV, TMV; Lysimachia vulgaris--ArMV, BNYVV, CMV, TMV; Lythrum salicaria- ArMV; Malva neglecta--CMV; Mercurialis annua--SoMV; Solanum nigrum--CMV, PVY, PVY(N); Solidago gigantea--CMV, RpRSV, BNYVV; Stenactis annua--PVM, PVA) weed- virus relations were detected. The epidemiological role of perennial hosts (A. syriaca, A. planlago aquatica, C. canurm, L. vulgaris, L. salicaria, S. gigantea) is especially high, because they can serve as infection sources as well as overwintering hosts of different plant viruses. PMID- 15759396 TI - Use of adjuvant-enhanced formulations to increase bipyridylium-herbicide effectiveness. AB - Paraquat and diquat are two popular, non-selective; bipyridylium herbicides commonly used in citrus orchards and horticultural row crops as the main chemical weed control method. However, since diquat lacks of an effective spectrum against grass weeds, and paraquat mammal toxicity raises strong environmental concerns, both an increase in diquat toxicity against grasses and a reduction in paraquat rates may be desired. Using grass-weed Lolium rigidum and broad leave weed Portulaca oleracea as experimental systems, the effects of six commercial adjuvants (poly-1-p-menthene, mixture of methyl oleate and palmitate, alkylglycol ester, dodecylbenzene ammonium sulphonate, and two paraffinic oils) on paraquat and diquat effectiveness have been studied under laboratory controlled conditions. Dose-response assays showed that adjuvants failed in increasing paraquat efficacy in both broad and grass weeds, yet antagonistic effects being observed in some mixtures such as paraquat + polymentene. However, all adjuvants tested did succeed in increasing significantly diquat effectiveness in P. oleracea and (most important) L. rigidum grass weed. Formulated-diquat ED50 rates were reduced down to 15% (diquat + DBSA) and 30% (diquat + fatty acid ester, diquat + polimentene) of those obtained on non-formulated-diquat trials for P. oleracea and L. rigidum, respectively. Results showed that formulated diquat proved to be a valid alternative to paraquat, and could be used as a more environmentally friendly substitute with comparable effectiveness and herbicide rate. PMID- 15759397 TI - Germination requirements and dispersal timing in two heterocarpic weedy asteraceae. AB - In SW Spain the winter annuals Anacyclus radiatus and Chrysanthemum coronarium (Asteraceae) are found as weeds in diverse crops. Both plant species are heterocarpic, i.e. the peripheral and central achenes of the capitulum are morphologically distinctive. In heterocarpic and heterospermic species the different fruit or seed morphs usually have differential ecological behaviour. In this work we have studied the morphometry, germination and dispersal timing of t he different achene morphs in A. radiatus and C. coronarium. Laboratory germination tests were carried out to evaluate the influence of incubation temperature (light/dark, 27/27, 24/18, 20/10, 15/5, 10/4 degrees C), chilling period (0, 1, 7, 21 and 60 days at 2 degrees C), GA3, nitrate and the dark on the germination percentage and rate (t50). The peripheral achenes of A. radiatus have a significantly broader wing than the central achenes. In this species, germination was inhibited in the dark but viable achenes of both types germinated completely under light exposure irrespective of test conditions. Nevertheless, the peripheral achenes germinated significantly faster compared to the central achenes (t5o 1.04 vs. 1.55 days at 24/18 degrees C). In C. coronarium peripheral achenes have three-winged ribs and are significantly longer, wider and thicker than the central achenes, which have only one- or two-winged ribs. In this species the peripheral achenes showed a much lower germination compared to the central achenes under all conditions tested (0.3-3.5% vs. 41.5-58.0%). Embryos isolated from the pericarp of peripheral achenes germinated quickly in a great extent, and when placed in close contact with a pericarp fragment they did not show any inhibition of germination, suggesting physical dormancy. To determine differences in dispersal timing between achene morphs, we monitored dispersal in a stand of both plant species. In A. radiatus the peripheral achenes were shed first, at the onset of the rainy season in late summer, whereas the central achenes dispersed from late summer to early winter. In contrast, in C. coronarium the central achenes were shed first, from early summer to early autumn, whereas the peripheral achenes dispersed mainly in early autumn. Thus, in both weed species the more dispersible and readier-to-germinate achene morph is dispersed first. The differences in the germination and dispersal behaviour between achene morphs represent an opportunistic, mixed strategy which is favourable under environmental uncertainty, and should be accounted for in order to understand the population dynamics of both weed species. PMID- 15759398 TI - Field experiences with recent ALS-inhibitors on herbicide resistant blackgrass (Alopecurus myosuroides Huds.). AB - In the growing season 2002-2003 two field experiments were carried out in winter wheat on the heavy clay soil of the coastal polder area at Zevekote to study the response of blackgrass (Alopecurus myosuroides Huds.) resistant or somewhat less sensitive to a wide variety of herbicides (clodinafop-propargyl, fenoxaprop-P ethy1; flupyrsulfuron-methyl+metsulfuron-methyl, propoxycarbazone-sodium; isoproturon) representing various modes of action. In Experiment 1, preemergence applications of isoproturon+diflufenican (1500+187.5 g/ha) and isoproturon+diflufenican+flurtamone (1250+100+250 g/ha) respectively were followed in mid-March (Zadoks: 23) by one of the following treatments: none, propoxycarbazone-sodium + vegetable oil (42 g/ha + 1 l/ha), mesosulfuron-methyl + iodosulfuron-methyl-sodium (+mefenpyr-diethyl) + vegetable oil (15+3 (+45) g/ha + 1 l/ha), clodinafop-propargyl (+cloquintocet-mexyl) {60 (+15) g/ha} and flupyrsulfuron-methyl+metsulfuron-methyl (10+5 g/ha). Systems based on clodinafop propargyl, propoxycarbazone-sodium or flupyrsulfuron-methyl+metsulfuron-methyl resulted in poor supplementary control of blackgrass compared to preemergence herbicide application only. On the contrary, systems based on postemergence application of mesosulfuron-methyl + iodosulfuron-methyl-sodium resulted in excellent control. In most cases the few surviving plants failed to produce inflorescences. In Experiment 2, fall applications in the 3 leaves stage (Zadoks: 13) of prosulfocarb + isoxaben (4000+75 g/ha), flufenacet + diflufenican + isoxaben (240+120+75 g/ha) and flufenacet + pendimethalin + chlorotoluron (180+900+1000 g/ha) respectively were followed in mid-March (Zadoks: 23) by one of the following treatments: none, propoxycarbazone-sodium + vegetable oil (42 g/ha+l l/ha), mesosulfuron-methyl + iodosulfuron-methyl-sodium (+mefenpyr diethyl) + vegetable oil {15+3 (+45) g/ha + 1 l/ha}, clodinafop-propargyl (+cloquintocet-mexyl) {60 (+15) g/ha}, flupyrsulfuron-methyl + metsulfuron-methyl (10+5 g/ha) and isoproturon+diflufenican (1000+125 g/ha). As in Experiment 1, systems based on clodinafop-propargyl, propoxycarbazone-sodium or flupyrsulfuron methyl+metsulfuron-methyl resulted in poor additional control of blackgrass compared to herbicide application in the fall only. Comparable poor levels of blackgrass control could be observed with isoproturon+diflufenican. Mesosulfuron methyl + iodosulfuron-methyl-sodium resulted in excellent control comparable to that recorded in Experiment 1. PMID- 15759399 TI - Evaluation of resistance in Cyperus difformis populations to ALS inhibiting herbicides. AB - Due to the intensification of rice cultivation in Spain and the repeated use of herbicides as an effective tool to control weeds, one important fact is the appearance of resistant populations of weeds, which are no longer controlled effectively at field doses. Therefore, it is necessary to increase doses to such an extent as to produce phytotoxicity in the rice crop. In paddy fields in Badajoz (Spain), a few years ago, populations of Cyperus difformis resistant to bensulfuron-methyl appeared. In order to characterize this resistance, whole plant assays were carried out in fourteen populations, eight of them originating from treated Spanish paddy fields. The results were expressed as resistance factor (RF) = ED50(R)/ED50(S). Two populations of Cyperus difformis, resistant (R) and tolerant (T) to bensulfuron-methyl, displayed resistance factors greater than 30 and 12.5, respectively. Furthermore, in order to evaluate the existence of a possible cross-resistance, the recommended field doses of 3 other herbicides (imazamox, ethoxysulfuron and bentazone + MCPA) were applied to the plants. All populations were susceptible to both imazamox and bentazone + MCPA and two populations showed cross-resistance to ethoxysulfuron. ALS activity was assayed in vitro in four biotypes, the R/S ratios of the I50 values calculated indicate that resistance involves a reduction in the sensitivity of the ALS mainly in the R population (biotype 4) to bensulfuron-methyl and ethoxysulfuron, displaying R/S ratios of 37500 and 142857, respectively, which may be attributed to inherent differences in the ALS protein itself and/or in the stability of the enzyme. PMID- 15759400 TI - Cross resistance to accase herbicide in Lolium rigidum. AB - Lolium rigidum is a cross-pollinating grass weed present in Europe and occurring in winter wheat and orchard crops. Several graminicides such as chlorotoluron and/or isoproturon and diclofop-methyl in mixtures or alone have been used successfully to control this weed in Spain during the past decade. However, several L rigidum populations have developed resistance to these herbicides following selection due to their continuous use. Four resistance mechanisms have been found in this grass weed, an enhanced metabolic detoxiflcation of the herbicides and an insensitive isoform of ACCase being the most important ones. The extent of cross-resistance depends on the type of mechanism. The biotype with an enhanced metabolic detoxification showed cross-resistance to ACCase-, ALS-, PSII- and tubuline-inhibiting herbicides, while the biotype resistant due to a mutation of the target site (ACCase) presented cross-resistance to ACCase inhibiting herbicides only. PMID- 15759401 TI - Evaluation of herbicide combinations for livid amaranth (Amaranthus blitum) control in tuberous begonia (Begonia x tuberhybrida). AB - In the past years livid amaranth (Amaranthus blitum) is observed increasingly in begonia production fields. Control of weeds in begonia is generally done by a combined application of the soil herbicides isoxaben + simazin followed 10 days later by application of the contact herbicide bentazone. This treatment usually controls the weed population sufficiently with exception of amaranth. In 2003 a field trial was conducted to evaluate control of livid amaranth in tuberous begonia with isoxaben, simazin. S-metolachloor, phenmedipham + desmedipham and bentazone. These herbicides were used as combinations of soil treatment and contact herbicides. The results suggest that a soil treatment of isoxaben + S metolachloor significantly reduces livid amaranth compared to isoxaben + simazin, without a pronounced negative effect on tuber yield. Application of phenmedipham + desmedipham however did not improve control of livid amaranth compared to bentazone. PMID- 15759402 TI - Effective control method of larvae of Diabrotica virgifera virgifera Leconte. AB - Larvae of WCR are feeding on the roots of corn while plants fall down. The egg hatching is continuous and soil insecticides are not effective to kill larvae. Unfortunately the recent control methods while we incorporate disinfectors Into the soil under seeding are not able to give enough effect on larvae of WCR under the whole period of larval development. We use to saw corn in the middle of April but eggs hatching start in the middle of May. The effectiveness of insecticides takes about one month so they are not able to protect plants from larvae are feeding on roots (Luckman et al., 1974 and Luckmann et al., 1975). They cause yield losses or in case of plant fall we can not harvest the corn. We have tested a material in greenhouse screening and field trips that is able to absorb insecticides and bind them into its body. This material is able to emit the agents continuously under the vegetation and we can protect our plants against the damages of WCR larvae. Our results shows that the material is able to elongate the effectiveness of the pesticides over 60 days and able to push the number of larvae under the economical threshold. PMID- 15759403 TI - Influence of the nonsteroidal ecdysone agonist, tebufenozide, on certain biological and physiological parameters of the cotton leaf-worm, Spodoptera littoralis (Boisd.) (Noctuidae: Lepidoptera) in Egypt. AB - In Egypt, cotton represents the most economical and important field crop. Cotton plant is the target of different pest species from cultivation until harvest. The Egyptian cotton leaf-worm, Spodoptera littoralis (Boisd.) (Noctuidae: Lepidoptera), is an important polyphagous insect pest attacking cotton, several cultivated crops and ornamentals worldwide. The integrated pest management (IPM) programs create the need for novel insecticides having more selective modes of action. So, the no steroidal ecdysteroid mimic, tebufenozide (RH-5992) was studied on some parameters of Spodoptera littoralis. This compound exhibits, in spite of its nonsteroidal structure, a high affinity for the ecdysteroid receptors (EcRs) through a lock-and key principle just like the natural insect molting hormone 20, hydroxyecdysone (20E). The present investigation was performed to throw more light on the following objectives: (1) the toxicity of tebufenozide against the fifth larval instar of a laboratory strain of S. littoralis, (2) the effect of this compound on some biological parameters, i.e. weight of mature larvae, mortality and percentages of larval malformations, (3) the action on some haematological parameters such as blood volume, total haemocyte counts (THCs), absolute haemocyte counts, differential haemocyte counts, and (4) the characterization of haemolymph proteins in treated as well as in the untreated larvae, using the electrophoresis technique on acrylamide gel expressed in the electrophoretic mobility. The obtained results could be summarized as follows: (1) the treated larvae reached their maximum weight after 48 h from treatment with tebufenozide; while the untreated ones reached the maximum weight after 72 h (2) the highest mortality (73%) occurred by applying tebufenozide at the concentration of 18 ppm. In addition, the deduced percentage of larval malformations, indicated that the maximum rate of malformed larvae was attained using the concentration of 36 ppm (3) the blood volume of the 5th instar larvae reached its maximum after 72 h from treatment; while in the untreated ones blood volume increased gradually and reached the maximum rate after 96 h (4) the total haemocyte counts (THCs) decreased in comparison with the control. The maximum value of THCs occurred at 72 h, resembling a lower rate (50%) than that of the untreated ones. (5) The absolute counts reached their maximum rates after 72 h from treatment. It was lower (80.7%) than the untreated larvae. (6) Tebufenozide cause the induction of (10.7 & 14.3 Kda) polypeptide in the haemolymph after 24 h of application. These bands were absent in that time in the untreated larvae (control). Later on, these bands were formed in the control larvae after 72 h of treatment. Induced synthesis of these transcript resulted in precocious molting. PMID- 15759404 TI - Effectiveness of imidacloprid (ProAgro 100 SL) in the control of glasshouse whitefly (Trialeurodes vaporariorum) and western flower thrips (Frankliniella occidentalis) on tomato under cover. AB - In 2002 a glasshouse experiments were carried out on the effectiveness of ProAgro 100 SL in the control of glasshouse whitefly and western flower thrips on tomato cv. Perkoz. Strict observation of the basic prophylactic rules such as introducing in to glasshouse only healthy seedlings, uninfected by whitefly and thrips as well as isolation of tomato plants, especially the seedlings from ornamental plants are important to the limit of the pest population. During the vegetation period the population of whitefly and thrips may be limited by the insecticide application. The glasshouse experiments on the performance of ProAgro 100 SL in the control of mentioned above pest species were carried out at the Research Institute of Vegetable Crops in Skierniewice. The ProAgro 100 SL was applied in the concentration of 0,1%. The standard treatment was Juventox 040 SL (acetamiprid) in the concentration of 0.05%. The obtained results allow to find that ProAgro 100 SL in the rates of 0.1% performed well in the limiting population of whitefly and western flower thrips on glasshouse tomato. PMID- 15759405 TI - Persistance of the insecticidal activity of five essential oils on the maize weevil Sitophilus zeamais (Motsch.) (Coleoptera: Curculionidae). AB - Essential oils of aromatic plants are popularise as protectant with low persistance. The evaluation of this duration of activity was carried out for five aromatic plants: Annona senegalensis (Annonaceae), Eucalyptus citriodora et Ecalyptus saligna (Myrtaceae), Lippia rugosa (Verbenaceae) and Ocimum gratissimum (Lamiaceae). They have significant insecticidal activity on S. Zeamais, on the first day of application, this activity decreases after 2 or 4 days. After 8 more than 50% of the efficacy is lossed for all the plants excepted A. senegalensis. PMID- 15759406 TI - Aphid specific predators in potato in Belgium. AB - In summer 2003, aphid specific predators populations were sampled in four potato fields located in Wallonia. Ladybirds, hoverflies and lacewings were the most abundant predators collected. A total of 6 ladybird, 8 syrphid and 2 lacewing species was found. Predator species composition and abundance greatly differed from field to field, while aphids populations were more or less similar with a peak of 3 to 5 aphid/potato leaf in mid-July. All chrysopidae sampled belong to the Chrysoperla kolthoffi (NAVAS) species except 2 specimens of Chrysopa perla L. Episyrphus balteatus DEGEER was the dominant syrphid species. Sphaerophoria scripta L. and Syrphus vitripiennis MEIGEN were subdominant species and 1 or 2 specimens of Melanostoma scalare F., Melanostoma mellinum (L.), Melliscaeva cinctella (ZETTERSTED), Metasyrphus corollae (F.) and Platycheirus albimanus F. were also collected. Coccinella septempunctata (L.) and Propylea quatuordecimpunctata (L.) were the dominant ladybird species. P. 14-punctata was proportionally more abundant in beating samples than with visual inspection samples, indicating that this ladybird is probably more discrete than C. septempunctata and that their populations can be underestimated with only visual inspection. Adalia bipunctata (L.) was also present in nearly all samples, but with lower levels of populations. Coccinella quinquepunctata (L.) and Adalia tenpunctata (L.) were found in small numbers at one occasion. Larvae and adults of Harmonia axyridis PALLAS, the multicoloured Asian beetle introduced in glasshouse for aphid control, were found in 3 fields out of 4, with 1.5 to 6% of ladybirds sampled. This is the first record of H. axyridis in open field in Belgium. As this beetle has been found in numbers at different locations and with different growth stages including larvae and pupae, it is supposed that H. axyridis is adapted to ware potato aphid and able to complete a full life cycle in this crop. As this species is very competitive for exploitation of food resources and aggressive for other predators, they are several questions about the evolution in the future of aphid specific predator biodiversity in ware potato. PMID- 15759407 TI - Amblyseius andersoni Chant (Acari: Phytoseiidae), a successful predatory mite on Rosa spp. AB - Roses on commercial nurseries commonly suffer from attacks by the two-spotted spider mite, Tetranychus urticae, which have a negative influence on growth and quality. The aim of this project is to find natural enemies that are well adapted to roses, and may improve biological control. At different sites such as a plant collection garden, public parks and field boundaries, leaves were sampled from roses to identify the indigenous species of predatory mites. Amblyseius andersoni was amongst other species frequently found, which suggests that this species thrives well on roses. The possibility for biological control of spider mites with A. andersoni was investigated both in container roses outdoors and in glasshouses. In plots of outdoor roses artificially infested with spider mites, the following treatments were carried out: spider mites alone (untreated plot), Amblyseius andersoni Amblyseius andersoni and ice plants, Neoseiulus californicus, Neoseiulus californicus and ice plants. There were four replications of the treatments. The ice plants, Delosperma cooperi, were added to some treatments to supply pollen as extra food for the predatory mites. Natural enemies such as Chrysoperla spp., Conwentzia sp., Orius sp., Stethorus punctillum, and Feltiella acarisuga occurred naturally and contributed to the control of spider mites. After one month the spider mites were eradicated in all treatments. At the end of the trial, predatory mites were collected from all plots for identification. The ratio of Amblyseius andersoni to Neoseiulus californicus was approximately 9:1. There was no obvious effect of the ice plants on the number of predatory mites. On a nursery, where new roses are bred and selected, Amblyseius andersoni was released in three glasshouses after one early treatment with bifenazate against two-spotted spider mite Tetranychus urticae. In two of these glasshouses Neoseiulus californicus was also released. Samples, which were taken in the summer months showed that the spider mites were kept at a very low level. Amblyseius andersoni was found, even if spider mites were absent. Rose plants infested with spider mites, that were brought in to the glasshouses later developed spider mite 'hotspots'. Phytoseiulus persimilis was introduced in the hot spots and contributed to the control along with Neoseiulus californicus, Amblyseius andersoni and naturally occurring Feltiella acarisuga. These observations showed that Amblyseius andersoni is a good candidate for preventing spider mite outbreaks, as it easily survives without spider mites. This predatory mite is able to survive on other food, including thrips and fungal spores. PMID- 15759408 TI - Effect of entomopathogenic fungus Tolypocladium species metabolite efrapeptin on Galleria mellonella agglutinin. AB - In the current study it has been found that efrapeptins, secondary metabolites of entomopathogenic fungi Tolypocladium cylindrosporum, interfere with agglutinin. The effect of efrapeptins on G. mellonella agglutinin was tested using rabbit blood. The results revealed that the end point for control larvae were 12.5 whereas for treated larvae which injected with 5 microl of 5 microg efrapepins ml(-1) or 50 microg efrapepins ml(-1) the end points were 10.5 and 8.5, respectively. Considering that efrapeptins suppress agglutination this study suggest that efrapeptins may interfere with the ligand-receptor interactions that are likely to occur at the plasma membrane of specific haemocytes. It has been suggested that majority of interactions between cellular and humoral components of the insect immune system are receptor-mediated. PMID- 15759409 TI - Drawing-up of pesticide selectivity lists to beneficial arthropods for IPM programmes in potato. AB - In order to promote IPM programmes in potato, the toxicity of 19 fungicides, 4 herbicides and 11 insecticides commonly used in this crop in Belgium was assessed on three beneficial arthropods. These species were representative of the most important aphid specific natural enemies encountered in potatoes: a parasitic wasp--Aphidius rhopalosiphi (De Stefani-Perez) (Hym., Aphidiidae), a ladybird- Adalia bipunctata (L.) (Col., Coccinellidae) and a hoverfly--Episyrphus balteatus (Dipt., Syrphidae). In a first time, pesticides were tested on glass plates on A. rhopalosiphi adults and A. bipunctata and E. balteatus larvae. For each insect, products inducing corrected mortality (Mc) lower than 30% were directly classified in a positive list for harmless products (green list). The other compounds were further tested on plants and listed in toxicity classes according to mortalities induced during this extended laboratory test: harmless (Mc < 30%), slightly harmful (30% < Mc < 60%), moderately harmful (60% < Mc < 80%) and harmful (Mc > 80). A chemical determination of pesticides residues was also performed for each experiment in order to determine the exposure of beneficial arthropods to pesticide residues and to validate the application of chemicals on tested substrates. On the basis of the results of acute toxicity tests, the period of each pesticide use according to normal agricultural practices and the abundance and importance of the three different groups of aphid natural enemies at different periods of the year, four pesticides lists were built up. Each list corresponded to a different period of pesticides application: Period I--from seedling to beginning of June (based on A. rhopalosiphi tests), Period II- beginning to end of June (based on A. rhopalosiphi tests), Period III beginning to end of July (based on E. balteatus and A. bipunctata tests) and Period IV- August to harvest (no exposure of beneficials). Results showed that herbicides were not toxic to the three species and can be used according to normal agricultural practices without restrictions. All fungicides can also be used without restrictions at recommended rates. Only the mixture Metalaxyl-M + Fluazinam was slightly harmful to A. bipunctata but had no effects on A. rhopalosiphi and E. balteatus. Results were more contrasted for insecticides and none of them was totally selective for all the 3 beneficial arthropods. Therefore, they can only be used with restrictions at periods II and III, according to the beneficial species that need to be protected. PMID- 15759410 TI - Cry1Ab protoxin effects on Spodoptera frugiperda (Lepidoptera: Noctuidae) and its parasitoid Cotesia marginiventris (Hymenoptera: Braconidae). PMID- 15759412 TI - Effect of Bt cotton on arthropod biodiversity in South African cotton fields. PMID- 15759411 TI - Transgenic-Bt potato plant resistance to the colorado potato beetle affect the aphid parasitoid Aphidius nigripes. AB - Using the biotechnological plant resistance for herbivore control with less reliance on chemicals in integrated pest management (IPM) programs critically depends on predictable interactions with no-target organisms of various trophic levels. Plant resistance to insect pests based on recombinant Bacillus thuringiensis could interfere with natural enemies of non target pests. Performance of the potato aphid parasitoid Aphidius nigripes was studied on the 'Superior-BT line transgenic for the CryllIA toxin of B. thuringiensis, resistance to the Colorado potato beetle; and none transformed 'Superior' line which served as control. Parasitoid survival was significantly lower on the 'Superior-BT' line compared to control. Adult females were largest on 'Superior' and smallest on BT potatoes. This difference was reflected on parasitoid fecundity, which was lowest on 'Superior-BT', and highest on Superior. The results indicate that factor of potato resistance to the Colorado potato beetle affected the fitness of a parasitold of the aphid Macrosiphum euphorbiae, a secondary pest of potato. The effects on the parasitoid were complex but were generally interpretable in terms of host aphid quality variation among potato lines used as food by the aphids during parasitoid development. PMID- 15759413 TI - Dissemination of the entomopathogenic fungus verticillium lecanii (Zimmermann) Viegas (Hyphomycetales: Moniliaceae) in a population of Frankliniella occidentalis (Pergande, 1895) (Thysanoptera: Thripidae). AB - Tests were carried out to investigate the dissemination of the entomopathogenic fungus Verticillium lecanii (Zimmermann) Viegas in a population of Frankliniella occidentalis. The tested factors, which influence the efficacy of the fungus against the pest insect, have been the population density of the thrips at the application moment as well as the temperature. The population density influenced the dissemination of the fungal spores in the population. The higher the density has been, the higher the insetting control effect has been as well. The temperature influenced the speed of the fungal effect, too. The higher the temperature has been, the earlier the control effect started. However, an increase of the natural mortality was found as well. PMID- 15759414 TI - Preliminary evaluation of the effectiveness of a Verticillium lecanii isolate in the control of Thrips tabaci (Thysanoptera: Thripidae). AB - Thrips tabaci is a polyphagous pest that attacks many different crop and ornamental plant species. Recently, the need to reduce the use of insecticides in the control of thrips is more and more increasing. On that account, the possibility to make use of the entomopathogenic fungus Verticillium lecanii (DAOM198499) was evaluated in laboratory conditions. Suspension of 10(3) to 10(7) conidia/ml grown in liquid culture medium with a control (distilled water) treated on second larval stage of T. tabaci in controlled condition (16L: 8D photoperiod, 25 +/- 1 degrees C temperature, 98% RH in first 48h and 70% RH in next days). Preliminary results showed that mortality in lethal suspension (10(7) conidia/ml) of V. lecanii after 24 hours and 2-7 days in lower suspension appears. This suggests that strine DAOM 198499 of V. lecanii has the ability to control T. tabaci in greenhouse conditions. PMID- 15759415 TI - Pathogenicity of the fungus, Verticillium lecanii, to the green peach aphid, Myzus persicae (Hom.: Aphididae). AB - Pathogenicity of the hyphomycete, Verticillium lecanii (DAOM 198499), was investigated to aphid, Myzus persicae under laboratory conditions. Analysis of lethal effect of six various concentrations 0, 10(4), 10(5), 10(6), 10(7) and 10(8) conidia/ml of V. lecanii against third nymphal stage of M. persicae, indicated significant mortality on aphids. Mean comparisons showed that there was significant difference between treatments. Three days after treatment, aphid mortality observed and after 12 days minimal mortality was 17.77 in control and maximum was 100 percent related to 10(7) and 10(8) conidia/ml. LC50 and LT50 values were estimated by probit analysis and life test. LC50 value for aphid mortality was 1.4 x 10(4) conidia/ml, LT50 values for concentrations 10(4), 10(6), 10(6), 10(7) and 10(8) conidla/ml was 10, 10, 9, 8 and 6 days, respectively. In this experiment, net reproduction rate of aphid's (R0) decreased significantly when concentration increased. These observations showed that V. lecanii (DAOM 198499) can be an active biological agent against aphids. PMID- 15759416 TI - Effection of Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) hoppers and adults by Beauveria bassiana (Deuteromycotina, hyphomycetes) conidia in an oil formulation. AB - Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) is one of the important insects pest in the heigh plains of north east of Algeria. Larval and adult stages caused severe damage to the leaves of cereals and an other various crops. Laboratory studies were conducted to the determine the potential of the fungus Beauveria bassiana for the control of the pest of larval and adults. Positive results were obtaved when larvae and adults were sprayed with different concentration of fungus conidia (D1: 2 x 10(6) conidia/ml, D2: 5 x 10(5) conidia/ml, D3: 8 x 10(4) conidia/ml, D4: 10(2) conidia/ml). Concentration at D1 and D2 in an oil formulation showed respectively 100% and 90 % larval mortality at the first day of treatment and 100%-9% adults mortality at the five day of treatement. The results revealed that the rates of infection and mortality were significantly related to the concentration of conidia in the corresponding samples. Beauveria bassiana can be considerd as a promising biocontrol agents of Ocneridia volxemi may be an other locuts, but field experimentation is necessary to subtantiate these finding. PMID- 15759417 TI - Bioassay of some Egyptian isolates of Bacillus thuringiensis against Culex pipiens (Diptera: Culicidae). AB - Some Egyptian isolates of Bacillus thuringiensis (BT) were grown on economic media contain 4% of fodder yeast in tap water and incubated under shaking conditions for four days. The biological activities of these isolates against Culex pipiens (Diptera: Culicidae) were carried out to determine their effectiveness against field and laboratory strains of 3rd larval instar. All isolates of BT were more pathogenic to laboratory strain. causing up to 84% larval mortality. The insecticidal activities of these isolates were extended to the pupal stage causing a significant effect on pupal mortality in both strains tested. A pronounced effect on adult emergence was noticed with remarkable adult malformations especially in the case of the isolate No. 2. The reproductivety of females was affected significantly by all isolates applied. PMID- 15759418 TI - Biodiversity of the microbial flora associated with two strains of Culexpipiens (Diptera: Culicidae). AB - Bacterial Flora from external surface and alimentary canal of wild and laboratory strains of Culex pipiens were isolated and investigated using quantitative bacterial cultures. Individual colonies were subcultured and identified to species level. Counts from alimentary canal differ significantly from those of the external surface. An increase in bacterial density was detected after feeding on mammalian and avian blood meal. Bacterial identification revealed a complex bacterial flora. In addition to members of family Enterobacteriaceae species of Bacillus, Staphylococcus, Streptococcus, and Acinetobacter are the most common in both strains investigated. Gram negative bacteria were increased significantly after feeding on blood meals than those detecting during the feeding on sugar after emergence of the adult female mosquitoes and vise versa with Gram positive bacteria. Bacterial isolates were tested for resistance to the most common commercial antibiotics. PMID- 15759419 TI - Laboratory evaluation of Melia azedarach L. and Eucalyptus globulus Labill. Extracts in order to control Ocneridia volxemi Bolivar (Orthoptera, Pamphaginae) hoppers. AB - This study was carried out to evaluate the effect of Melia azedarach L. (Meliaceae) dry fruit extracts (oil and water) and Eucalyptus globulus Labill. (Myrtaceae) dry leaves extracts (water) on Oneridia volxemi hoppers (fifth instar) feeding on cereals leaves (Triticum durum) under laboratory conditions. Larva feeding reduced and mortality was significatly (p < 0.05) on cereals leaves sprayed with three concentrations 0, 5, 1 and 2% of oil extracted from Melia azederach. In addition, O. volxemi feeding and mortality was significantly (p < 0.05) on cereals leaves sprayed with two concentrations 50 g/l and 80 g/l of water extracts from Melia azedarach dry fruits soaked for 24 hours. the water extract solution of 80 g/l significantly reduced feeding than the other concentrations 50 and 25 g/l. This study showed also the water extract solution from Eucalyptus globulus dry leaves reduced also feeding and induced the mortality of O. volxemi but not better than Melia azedarach dry fruit extracts water at the same concentration (80 g/l). The aim of this study is in an integrated management program for control of O. PMID- 15759420 TI - Antifeedant activity of Cestrum parqui and Drimys winteri on Spodoptera littoralis (Lepidoptera: Noctuidae). PMID- 15759421 TI - Effects of flucycloxuron, a chitin synthesis inhibitor, on reproductive events and thickness of chorion in mealworms. AB - Flucycloxuron (FCX), a benzoylphenylurea derivative, was evaluated on Tenebrio molitor. The compound was incorporated into the diet and administrated to newly emerged females at various doses (2, 5 and 10 mg/kg). FCX was found to affect several reproductive events such as the duration of preovipostion and oviposition period, the fecundity, the viability of eggs and the duration of embryonic development, respectively. Morphological study of ovaries showed that FCX reduced both oocytes number, the ovaries weight and the size and the volume of the basal oocyte during the sexual maturation. In addition, it reduced the thickness of chorion from freshly laid eggs. However, electron microscopic study revealed that this compound had no significant effect on the fine structure of chorion. Finally, measurements of ovarian ecdysteroids production by an enzyme immunoassay indicated a reduction in the hormonal amounts recorded. PMID- 15759422 TI - Reproductive effects in German cockroaches by ecdysteroid agonist RH-0345, juvenile hormone analogue methoprene and carbamate benfuracarb. AB - Blatta germanica is the more prevalent cockroach species in Algeria. In the present study, we tested the effect on reproduction in B. germanica of two insect growth regulators, RH-0345, a benzoylhydrazine analogue that mimics the action of 20-hydroxyecdysone, and methoprene, one of the most commercially important juvenile hormone analogues, and a novel carbamate insecticide, benfuracarb. The compounds were applied topically (10 and 20 microg/insect for RH-0345, and 1 and 10 microg/insect for methoprene) or orally administrated (at 2% for benfuracarb) on newly emerged females and evaluated on reproductive events during the adult life (2, 4 and 6 days). Treatment with RH-0345 and benfuracarb reduced significantly the number of oocytes, the size and the volume of the basal oocyte during the experimental period. Methoprene distorted the ovarian development since it caused a significant reduction in the number of oocytes at 2, 4 and 6 days for the two tested doses, and an increase in oocyte size at 2, 4 and 6 days with 1 microg and a decrease with 10 microg. In a second series of experiments, the effects of these compounds were assayed on the ovarian proteins. Data from biochemical analysis revealed that RH-0345 and benfuracarb reduced the ovarian amounts of proteins, while treatment with methoprene increased it during the sexual maturation. PMID- 15759423 TI - Investigation on the optimal term of one-fold insecticide application for decreasing onion thrips ( Thrips tabaci Lindeman, Thysanoptera, Thripidae) damage on early white cabbage. AB - An impact of the term of one-fold insecticide application to reduce the damage of the onion thrips in early white cabbage was established. During the growing season plants were treated with insecticide (abamectin) in three different terms (treatment 1: 9 June, treatment 2: 16 June, treatment 3: 23 June, and treatment 4: untreated). On the exterior leaves of cabbage heads, treated with the insecticide, statistically significantly lower mean index of damage was determined as compared to the untreated plants. No significant differences were found between three different terms of application. The majority of the economically important damages in all of the treatments was found between the 3rd and 6th exterior leaf in the head. The highest mean weight of heads (1517.3 g) and mean net weight of heads I = mean weight of heads - weight of damaged and removed leavesl (1166.3 g) were established in plants which were treated the last. These parameters were the lowest in untreated plants (1083.3 g / 805.6 g). The yield loss due to damaged leaves removal amounted from 22.9% (treatment 2) to 25.6% (treatment 4). Based on the results obtained in this research we concluded that yield loss due to onion thrips attack in plants with one-fold insecticide application is not substantially lower as in untreated plants (though statistically significant differences between them were established), because insecticide cannot reach the interior leaves in the head. Substantial differences in the total and net weight of heads between the treatments and especially between the treated and untreated plants are explained by the fact that feeding of numerous thrips populations in the heads and on the exterior cabbage leaves negatively affects plant physiology and yield. The highest average yield in plants which were treated the last indicates a possibility that insecticide also inhibits plant growth and development to a certain extent. PMID- 15759424 TI - Seasonal occurrence and relative abundance of aphids on potato plants with classical and transgenic characters of resistance to Colorado potato beetle Leptinotarsa decemlineata (Say). AB - The seasonal abundance and dynamics of potato-infesting aphids were studied in two seasons, on potato Solanum tuberosum L. with various types of transgenic and classical resistance to primary pests, especially the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say). The lines tested were the NewLeaf potato, a transgenic expressing the Bacillus thuringiensis CryIIIA toxin (BTT); NYL 235-4 a parental line derived from S. berthaultii (NYL) with a high density of glandular trichomes; and a transgenic line expressing oryzacystatln I (OCI), a proteinase inhibitor gene from rice; potatoes from two commercial cultivars, Superior and Kennebec, served as controls. Over the two seasons, Macrosiphum euphorbiae (Thomas) and, to a lesser extent, Myzus persicae (Sulzer) were by far the most abundant aphids observed in the experimental plots. M. persicae was observed relatively late in the season, and was most abundant on the Kennebec-OCI potato. Macrosiphum euphorbiae was more regular over seasons, and its highest densities were observed on the NYL 235-4 line. The population of M. euphorbiae showed markedly different patterns on the five lines studied, which was clearly related to potato plant phenology. Density of M. euphorbiae was observed on the NYL 235-4 line, in late July and early August. PMID- 15759425 TI - Colorado potato beetle larvae on potato plants expressing a locust proteinase inhibitor. AB - The natural defence system of plants often involves inhibitors of digestive enzymes of their pests. Modem and environmental-friendly methods try to increase this plant resistance by expressing heterologous protease inhibitors in crops. Here we report the effects of expressing a gene from desert locust (Schistocerca gregaria) encoding two serine protease inhibitors in potato on Colorado potato beetle (Leptinotarsa decemlineata) larvae. The gene encoding both peptides on a single chain was used for Agrobacterium-mediated transformation of potato plants. The presence of the active inhibitor protein in the leaves was verified. The feeding bioassays in the laboratory showed that despite the low level of the peptide in leaves, CPB larvae on transgenic plants have grown slightly but significantly more slowly than those on control potato plants. The results support the notion that expression of multifunctional proteinase inhibitors of insect origin in plants might be a good strategy to improve insect resistance. PMID- 15759426 TI - First report of the western corn rootworm Diabrotica virgifera in Belgium. PMID- 15759427 TI - Action of some micronutrients on the infestation and yield components of faba bean by the aphid, Aphis craccivora Koch (Aphididae, Homoptera) and the leaf miner, Liriomyza trifolii (Burgess) (Agromyzidae, Diptera). AB - Field experiments were carried out in the two growing seasons of 1999/2000 and 2000/2001 on faba bean (Vicia faba) plants in the Experimental Farm of Agriculture Research Station at Nubaria region, Alexandria, which is considered as a newly reclaimed calcareous soil. The present investigation aimed to evaluate the effect of spraying faba bean plants with certain micronutrients, i.e. Iron, Manganese and Zinc either in single double or triple combinations on the infestation by the aphid, Aphis craccivora Koch (Aphididae, Homoptera) and the leaf miner, Liriomyza trfolu (Burgess) (Agromyzidae, Diptera). The infestation by these insects was assessed using the parameters of Infestation grades as well as the injury indices. Faba bean plants cv. Giza Blanca were sprayed twice (45 and 66 days) after planting with the above-mentioned micronutrients. However, results of this investigation showed, with no doubt, that Mn, Zn and Fe individually or in double or triple combinations have increased to varied extents the infestation rates (%) of faba bean plants compared to the untreated ones. Such varied increases were mainly due to the metabolic roles of the used foliar sprays and their interactions, which indirectly affect the physio-biological actions of plants that may render them suitable for either A. craccivora or L. trifoii reproduction. This phenomenon might be also due to the different environmental factors. In both seasons, the relationship between nutrients applications and pests Infestation followed the same trend of increase in the percentages of infested plants. This assures and confirms the constant metabolic roles of such micronutrients. The biological seed weight (ton/fed.) was positively affected by the application of the used micronutrients. It is worth mentioning that the maximum response was observed in case of the triple treatment followed by the double and single treatments in a descending order. Application of the investigated micronutrients alone or in mixtures resulted in significant increases in yield and its components. Such increases were due to the fact that ions of Zn, Fe and Mn are cofactors of several enzymes, but rarely if ever with a high degree of specificity. PMID- 15759428 TI - The ecological study of cotton bollworm (Helicoverpa rmigera Hubner 1808) in Hungary. AB - The cotton bollworm (Helicoverpa armigera Hbn.) is a poliphagous pest. Caterpillars feed on flowers, crops and seeds. In 2001 the meaningful catching period was in August (Szeoke, 2001). In 2003 we detected the swarming already in June. We observed many caterpillars on its nutritive crops. It caused significant economic damage in this year. 1ST EXAMINATION: We collected larvae and reared pupae out of it in a pot. We took it into the soil. The swarming of the moths from the pots was in June. The mortality was high, more than 90%. 2ND EXAMINATION: We made cold tests with pupae. We examined 5 x 10 pupae in three treatments. In the first treatment we reduced the temperature to -2 degrees C for 4 weeks. 92% of the pupae survived this cold. In the second treatment we reduced the temperature to -2 degrees C for 3 weeks and to -7 degrees C for 1 week. 86% of the pupae survived this procedure. In the third treatment we reduced the temperature to -2 degreesbC for 3 weeks and -15 degrees C for 1 week. 100% of the pupae were perished. 3RD EXAMINATION: In the first treatment we raised caterpillars on 13 hours lighting and 24 degrees C. The swarming was from 20th April to 4th May 2004. In the second treatment we reared the worms on 20 hours lighting and 18 degrees C. The main swarming was on 3rd January 2004. So we could say that the cotton bollworm has diapause. The more effective factor of the diapause is the length of the lighting. PMID- 15759429 TI - Plant wounding affects the oviposition of Hellula undalis(F.). The impact of leaf chemistry. AB - Crucifer specialists, such as the tropical pest Hellula undalis (F.), developed a close association to their hosts and depend on characteristic secondary plant compounds, the glucosinolates (GS), in many cases. Following tissue damage, GS undergo hydrolysis catalyzed by the enzyme myrosinase to produce a complex array of products which include isothiocyantes, cyanides, and thiocyanates depending on reaction condition and the chemical nature of the side chain. We found that mechanical plant damage to the tropical crop plant Brassica campestris L. ssp. chinensis (pak-choi) leads to increased GS content, especially of indolyl GS and affected the oviposition behaviour of H. undalis negatively. GS were detected in methanolic leaf surface extracts from intact B chinensis and Sinapis alba (L.) plants, but no hydrolysis products of GS were found in leaf extracts with methylene chloride. Furthermore, the oviposition stimulant activity of GS extracts for H. undalis was greatly reduced after hydrolysis of GS by myrosinase. These results suggest the importance of GS for inducing oviposition, thus excluding hydrolysis products as active principle. Specific produced slightly volatile hydrolysis products of GS (3-indolylmethyl-cyanide and 5 vinyloxazolidine-2-thione) reduced oviposition, when applied to the host-plant. Leaf chemistry, including volatile and non-volatile compounds, may determine oviposition in H. undalis indicating a suitable host-plant. PMID- 15759430 TI - Seasonal abundance of two armyworm species, Spodoptera exitgua (Hubner) and Spodoptera litura (F.) in the Philippines. AB - The two armyworm species Spodoptera exigua (Hubner) and S. litura (F.) are Important pests on several agricultural crops in the tropics. Knowledge about species abundance is important for the development of integrated pest management strategies. During 1998 and 1999 population densities were recorded weekly using pheromone traps at San Leonardo and Munoz (Province Nueva Ecija, 150 km north of Metro Manila) on the Philippine island Luzon. Furthermore, meteorological conditions and natural enemies were observed. Both armyworm species were present year-round at both observation sites. No differences in numbers of male insects trapped in pheromone traps between seasons were found for S. litura. Contrary, numbers of male S. exigua trapped during rainy season was significant higher than during dry season. Rainfall explained between 30 and 40% of trapped moths and has to be considered when monitoring population density with pheromone traps. Parasitism rates were below 5% in both years and for both species and can not explain the high variation observed in population densities. PMID- 15759431 TI - Rearing of the cucurbit fly Dacus cilratus Loew (Dip: Tephritidae) on artificial diet under laboratory conditions. AB - Cucurbit fly is an important pest of cucurbit plants (Cucumber, melon and watermelon) in most mid-east countries including, Iran. This insect causes a high damage so that several sprayings are required to control the pest. Using male strile is another physical method for controlling this pest. For this purpose it is necessary to rear many male insects. Therefore an investigation was carried out to compare different artificial diets for rearing the melon fly. First, pupae were collected from cucurbit field and transferred to laboratory. In this experiment five diet formulations were compared and the following was used for rearing: Wheat bran (14 g), soybean lees (3 g) sugar (50 g), yeast extract (2.7 g) nipagin (0.1 g). sodiumbenzoate (0.1 g) Hydrochloric acid 3.5% (4.0 ml) and distilled water 70.5 mil. For rearing the adult, the best diets contained: Brower yeast (1 part), Honey (5 parts), water (94 parts) and a slice of cucumber. These diet caused abundance of oviposition and egg fertility. PMID- 15759432 TI - Species composition of leaf beetle assemblages in the canopies of apple and pear orchards in Hungary and Great Britain (Coleoptera: Chrysomelidae). AB - The species richness and species composition of Coleoptera assemblages were investigated in the canopies of apple and pear orchards in Hungary and in the apple orchards in Great Britain. The investigations were carried out in Hungary (Nagykovacsi: 3 plots, Kecskemet: 5 plots, Sarospatak: 4 plots) between 1990-94, and in Great Britain in Kent (East Malling, Marden and Robertsbridge) in 2001 and 2002. Former investigations in Hungary revealed that the diversity of Coleoptera assemblages in the canopy of apple and pear orchard were surprisingly high. As a result of our investigations it was found that altogether 324 species, almost 3% of the Hungarian beetle fauna were represented: 253 species in apple orchards and 188 species in pear orchards. Similar results were obtained in the investigations carried out in Great Britain between 2001-2002. In Hungary, the majority of the species belonged to the families Curculionidae, Chrysomelidae and Coccinelidae. The proportion of leaf beetles varied between 15 and 20%. The most common leaf beetle species in the canopy of the commercial orchards were Phyllotreta vittula, Phyllotreta atra, Phyllotreta nigripes, Oulemta melanopus, and Aphthona euphorbiae. In the abandoned orchards the most common species were Luperus xanthopoda, Smaragdina salicina and Orsodacne liieola. In Great Britain 44 leaf beetle species were found in the canopies of the investigated orchards. The species with higher abundance were Aphthona euphorbiae. Chaetocnema concinna and Longitarsus parvulus. We concluded, that leaf beetles give high part of the orchard canopy biodiversity and sometimes occur with high species richness and abundance. However, the reasons of their occurrence and their potential role are poorly known. PMID- 15759433 TI - First report of Eotetranychus fagi in Belgium. AB - Eotetranychus fagi (Acari: Tetranychidae) was first recorded in Belgium on Fagus sylvatica in Kortrijk in October 2002. In the autumn of 2003 E. fagi was noticed again at several locations in Flanders. Because F. sylvatica is often used as hedge plants in private gardens, it is expected that further spread of this spider mite will occur in the next few years. PMID- 15759434 TI - Movement of entomopathogenic nematodes in soils of Fragaria spp. AB - The mobility of three entomopathogenic nematodes (Steinernema carpocapsae UK and All Strain, and S. glaseri (Nematoda: Steinernematidae) was observed in strawberry fields in Vancouver, WA. Nematodes were sprinkled over the soil surface using a mason jar. Soil and soil-moisture content, soil composition, soil pH and conductivity was reported over the evaluation period. 12 cm deep soil samples were taken after surface application at 9 and 25 days, 7, 26 and 45 and 6, 22, 37 and 50 days at the three different test sites within the surface application site and at 5 cm distance. Soil samples taken were divided into three layers (0-4, 5-8 and 9-12 cm) and exposed once to five Galleria larvae in the laboratory to evaluate nematode presence and movement. Results after surface application demonstrated more vertical movement of S. glaseri into deeper soil layers in comparison to both S. carpocapsae strains. PMID- 15759435 TI - Use of high resolution digital thermography to detect Heterodera schachtii infestation in sugar beets. AB - Thermography is a non-destructive method used to monitor pest and disease infestations, as it is related to changes in plant water status. Surface temperature differences of the crop canopy may be an indicator of nematode infestation as the parasitation of the root system reduces evaporation of leaves. To test the potential of high resolution digital thermography to detect Heterodera schachtii infestation, experiments using increasing nematode densities and different sugar beet varieties were conducted. From June to August 2003 the crop canopy temperature was measured with a thermal infrared camera from a helicopter. A significant correlation between canopy temperature and nematode density was observed with the susceptible cultivar Monza whereas the resistant cultivar Paulina did not show any correlation. Mean temperature comparison showed significant differences between the lowest infestation level (500 eggs and larvae/100 ml soil) and the highest infestation level (>1500 eggs and larvae/100 ml soil). At the beginning of the season canopy temperature differences between healthy and nematode infested sugar beets were higher (approximately 1 degree C) compared to later assessment dates when the water supply in the soil was limited. Since low and high nematode infestation could be clearly distinguished with the susceptible cultivar by airborne thermal images, thermography might be a useful tool for monitoring sugar beet fields. PMID- 15759436 TI - Efficacy of Paecilomyces lilacinus (strain 251) for the control of Radopholus similis in banana. AB - Paecilomyces lilacinus is a common soil fungus that has been isolated from many different habitats around the world. It is well known as a facultative egg pathogen of sedentary nematodes and also an important option to control Radopholus similis juvenile and adults in banana. This nematode antagonistic fungus may be used in an integrated approach to control banana plant parasitic nematodes. Dose response and form of application experiments were conducted with burrowing nematode, R. similis, on banana using a commercial water dispersible granulate formulated P. lilacinus (strain 251) product. The results revealed that nematode activity decreased in the presence of this fungus. An important correlation between rates of application and the degree of control of R. simnilis penetration and banana root weight was observed. The best control was achieved in the treatment were plantlets and soil were pre-inoculated with P. lilacinus and reinoculated during transplantation. The results showed that the biocontrol agent P. lilacinus is an excellent candidate for an IPM program against nematodes such as Radopholus similis. PMID- 15759437 TI - Optimizing the efficacy of Paecilomyces lilacinus (strain 251) for the control of root-knot nematodes. AB - The egg pathogenic fungus Paecilomyces lilacinus (strain 251) is a biocontrol fungus with a potential range of activity to control the worldwide most important plant parasitic nematodes. This biological nematicide may be an useful tool in an integrated approach to control mainly sedentary nematodes. Greenhouse experiments were conducted with the root-knot nematodes Meloidogyne incognita and M. hapla on tomato. P. lilacinus, formulated as WG (BIOACT WG), was incorporated into soil inoculated with root-knot nematode eggs prior to transplanting the susceptible tomato cultivar "Hellfrucht". Furthermore, soil treatments were combined with seedling treatments 24 hours before transplanting and a soil drench 2 weeks after planting, respectively. Seedling and post planting treatment was also combined with a soil treatment at planting. All single or combination treatments tested decreased the gall index and the number of egg masses compared to the untreated control 12 weeks after planting. However, the combination of the seedling treatment with a pre- or at-planting application of P. lilacinus was necessary to achieve higher levels of control. Additional post plant drenching resulted in only a slight increase In efficacy. To the feasibility of this modified application system for the control of root-knot nematodes, a yield experiment was conducted with M. hapla and the susceptible cultivar "Gnom F1 Hybrid". It could be demonstrated that the above mentioned combination of pre-planting application plus the seedling and one post plant drench gave the best control and resulted in a significant fruit yield increase in concurrence with a decrease in number of galls per root. PMID- 15759438 TI - Effects of endophytic Fusarium oxysporum towards Radopholus similis activity in absence of banana. AB - Four endophytic fungi (Fusarium spp.) isolated from the cortical tissue of surface-sterilised banana as well as from tomato roots were tested for their capacity of biological control towards the burrowing nematode Radopholus similis on banana. The pathogenic and parasitic capacities of endophytic fungi towards R. similis were tested in in vitro experiments. No parasitism of fungi on R. similis was observed. However, nematode activity decreased significantly in the presence of all endophytic fungi in vitro when compared to nematodes in the absence of fungi. The effects of fungi on R. similis activities in the soil were tested in the absence of plants. Nematode activities were reduced significantly by 16-30% by endophytic fungi when compared to untreated soil. PMID- 15759439 TI - Interactions between root-knot nematode Meloidogyne javanica and Fusarium wilt disease, Fusarium oxysporum f.sp. Melonis in different varieties of melon. AB - Fusarium oxysporum f.sp. melonis and root-knot nematode (Meloidogyne javanica) are destructive pathogens on cucurbits in Varamin area of Iran. The interaction between two pathogens was studied on local melon cultivars, Garmsar and Sooski. Inoculum of Meloidogyne javanica was prepared on susceptible cultivar, Rutgers using single egg mass method in greenhouse. Inoculum of Fusarium oxysporum f.sp. melonis (race 1) was prepared using Richard solution. A concentration of 2 x 10(5) micro conidia of fungus and 2000, 3000, 4000, 5000 eggs of nematode was used in 1 kg of autoclaved soil. Plants were inoculated with nematode at 2-3 leave stage then with fungus 2 weeks after nematode inoculation. The experiment was conducted in factoriel design based on CRD with 20 treatments, including varieties in 2 levels (Garmsar and Sooski), nematode in 5 levels (0, 2000, 3000, 4000, 5000 eggs) and fungus in 2 levels (presence and absence) and 3 replicates. The index that evaluated were growth index including fresh and dry weight of shoot and root, height, Fusarium wilt index and root gall index. Results of this experiment showed that all of treatments comparison to control were significantly different (p = 0.05) in growth index. Combination of fungus and nematode (5000 eggs) caused the most decrease in growth index on Garmsar and Sooski. PMID- 15759440 TI - Effect of tea dust residues to control root-knot nematode of tomato. AB - In this research, control of tomato root- knot nematode (Meloidogyne incognita) was conducted using tea dust residues at different rates. First, the species and race of nematode were identified by employing diagnostic keys. Then, with 5 replications in complete randomized design. Tea dust residues were used at 9 treatments (0, 5, 10, 15, 20, 25, 30, 35 and 40 g/kg of soil). Statistical analysis on mean treatments rates showed that treatment with 25 g/kg soil economically was effective in growth rates and reduction in gall index. PMID- 15759441 TI - Occurrence of plant parasitic nematodes (Tylenchina) in sugar beet fields in Fars province, Iran. AB - On a survey of plant parasitic nematode fauna, belonging to the suborder Tylenchina, about 110 soil and root samples were collected from different sugar beet field in fars Province in Iran. The samples were washed and the nematodes were extracted by Centrifugal Flotation Technique according to the modified method by De Grisse, 1969. They were fixed and transferred to glycerine, then permanent slides were mounted. Morphological and morphometrical characters of collected species were carefully studied and the characters were compared with the original descriptions and differences were discussed. In this study 18 species belonging to 11 different genera of subordes Tylenchina were identified as follows. Boleodorus thylactus, Ditylenchus destructor, D. dipsaci Geocenamus brevidens, G. rugosus, Helicotylenchus digonicus, H. pseudorobustus, H. vulgaris, Heterodera filipjevi, H. schachtii, Pratylenchus neglectus, P. thornei, Psilenchus hilarulus, P. hilarus, Stictylus mucronatus, Tylenchorhynchus ventrosinatus, Tylenchus davainei and Zygotylenchus guevarai. The species of the genera Ditylenchus, Heterodera, Pratylenchus and Psilenchus were comparatively more distributed than the others. This investigation revealed that sugar beet cyst nematode (Heterodera schachti) was found already in most sugar beet fields in this Province. Fathabod, Marvdasht and Tasuj-kwar were the most infested areas. Cysts collected from 100 gr. Of soil samples were 120 to 121 in number and the eggs and second stage larvae in each gr. of soil were 104.1 to 104.5 respectively. PMID- 15759442 TI - [Improvement of outpatient therapeutic care is a topical problem of health care]. PMID- 15759443 TI - [Pneumonia: errors in outpatient diagnosis and treatment]. AB - AIM: To reveal errors in diagnosis and treatment of pneumonias in outpatients. MATERIAL AND METHODS: The analysis of 345 case records of outpatients. RESULTS: It is demonstrated that initially most of the patients had no symptoms and physical data pointing to pneumonia. This may be one of the causes of late diagnosis (29.7%). The first visit to the doctor brought out misdiagnosis--acute respiratory viral infection (ARVI)--in 96.6% cases. The choice of starting antibacterial therapy did not correspond to recommendations of the Standards of Diagnosis and Treatment of Patients with Nonspecific Diseases of the Lungs in 22.8% cases. 20.6% patients of this group demanded change of antibiotics. Unnecessary long-term therapy had place in 11.5% patients. 24-h underdose of antibiotic was given in 18.6%, inadequate dose regimen was registered in 5.6%. Instead of 13.3% patients, 51.3% were hospitalized. Control x-ray examination was performed in 75.9%. Lesions in the lungs remained in 36.5% patients. CONCLUSION: Quality of pneumonia diagnosis and treatment in outpatient setting must be updated. PMID- 15759444 TI - [Informative value of exercise test for prognosis: case report of 20-year follow up of a patient with ischemic heart disease (lecture)]. PMID- 15759445 TI - [Current clinical picture of primary subacute infectious endocarditis and its prehospital diagnosis (lecture)]. PMID- 15759446 TI - [Outpatient follow-up and management of pregnant women with hypertension]. AB - AIM: To study variants of the course of chronic hypertension in pregnant women and determination of factors predisposing to a persistent rise of arterial pressure (AP) in pregnancy. MATERIAL AND METHODS: A total of 50 pregnant women were examined (heart rhythm variability and psychological testing) who had AP 140/90 mm Hg and higher as shown by measurements at three outpatient check-ups. After delivery the patients were retrospectively devided into two groups. Twenty seven group 1 women had frequent rises of AP to 140/90 and higher throughout pregnancy; twenty-three women of group 2 had high AP only at early terms of pregnancy, later they became normotensive without use of hypotensive drugs. 24-h AP monitoring was made in 29 patients. By its results, two subgroups were identified: 11 patients with essential hypertension and 18 women with neurocirculatory dystonia by hypertensive type. RESULTS: In group 1 there was an early fall of cardiac performance, higher values of SMIP test according to scales 2 (pessimism), 3 (emotional lability), 4 (impetuosity) and 7 (anxiety). CONCLUSION: The analysis of 24-h AP profiles revealed more persistent and significant rise of AP in patients with essential hypertension than in those with neurocirculatory dystonia. They also demonstrated high AP variability correlating with a risk of cardiovascular diseases. PMID- 15759447 TI - [Differential approach to detection and treatment of cholelithiasis in outpatient conditions]. PMID- 15759448 TI - [Day hospital: experience and efficacy]. AB - AIM: To analyse efficacy of treatment in a day hospital (DH) of Avtotrans (Orenburg city) outpatient department. MATERIAL AND METHODS: The activity of the DH was assessed by case histories and annual records for 5 years. RESULTS: DH provides high-quality examination and treatment for patients with different diseases (efficacy of treatment 99.5%). Ulcer treatment analysis showed that 98.53% of DH ulcer patients achieve a complete healing of the ulcer defect and its decrease was observed in 1.47%. CONCLUSION: DH facilities are able to provide differential therapy of patients and active prophylaxis of chronic disease progression. Combined therapy under a close control of physicians results in a good therapeutic effect. PMID- 15759449 TI - [Principles of expert evaluation of disability in visceral diseases in outpatient clinic]. PMID- 15759450 TI - [Thyroid diseases in reproductive-age women living in Shoria mountains]. AB - AIM: To analyze prevalence and structure of thyroid pathology in women of reproductive age living in Shoria mountains. MATERIAL AND METHODS: The examination of 409 women aged 18 to 40 living in mountain Shoria included assessment of social status, heredity, actual nutrition, visit to endocrinologist, ultrasound investigation of the thyroid, tests for thyroid hormones (TTH, T3, T4) in the serum, antibodies to microsomal fractions, iodine concentrations in the morning urine. RESULTS: Significant prevalence of thyroid diseases (62.3%), high rate of diffuse nontoxic goiter of the second degree (18.0%), nodular nontoxic goiter (11.8%), autoimmune thyroiditis (6.3%), hypothyroidism (5.5%), low median of iodine excretion with urine in women refer mountain Shoria to the zone of goiter endemia. CONCLUSION: Among migrants thyroid morbidity is significantly higher than among the native population. Eradication of goiter endemia will promote reproductive health in this region. PMID- 15759451 TI - [Major cardiovascular diseases mortality in Kaunas population for 1983-2002]. AB - AIM: To evaluate trends in mortality of ischemic heart disease (IHD) and stroke in Kaunas population aged 25-64 years in 1983 to 2002. MATERIAL AND METHODS: The study based on official mortality statistics covered all residents of Kaunas aged 25-64 years who died of IHD and stroke in 1983-2002. Age-standardized rates were calculated by the direct method and using European population as a standard. The trends were analysed using the method of linear regression on logarithms of the age-standardized annual rates. RESULTS: In 1983-2002, IHD and stroke mortality rates were significantly decreasing both for men and women (by 2.2%/yr, p = 0.003, and 2.9%/yr, p = 0.004, respectively, for men and by 2.6%/yr, p = 0.005 and 3.2%/yr, p = 0.002, respectively, for women). CONCLUSION: In 1983-2002, both IHD and stroke mortality rates among both Kaunas men and women aged 25-64 years decreased significantly. PMID- 15759452 TI - [Alcohol consumption and mortality among kaunas population aged 35-64 years from 1983 to 2002]. AB - AIM: To evaluate time trends in alcohol consumption habits and alcohol-related mortality among Kaunas (Lithuania) population aged 35--64 years from 1983 to 2002. MATERIAL AND METHODS: The WHO MONICA study was carried out in Kaunas from 1983 to 2002. Alcohol consumption habits were determined using the data of four surveys among random samples of population aged 35-64 years (overall 3292 men and 3548 women, the response rate 65.1%). The source of mortality data is official mortality statistics of all permanent residents of Kaunas aged 35-64 years in 1983-2002. Age-standardized rates were calculated by the direct method and using World population as a standard. The trends were analyzed using the method of linear regression on logarithms of the age-standardized annual rates. RESULTS: Over the 20 years, the prevalence of regular alcohol consumption significantly increased among both men and women. At the same time, there was an increase in the study population of a mean level of consumed alcohol and the proportion of drinkers consuming alcohol frequently. During 1983 to 2002 mortality rates from external causes and suicides were increasing significantly among men (by 2.1%/yr., p = 0.039 and by 3.4%/yr., p = 0.0002, respectively) and have not changed among women. CONCLUSION: Negative trends in alcohol consumption habits and alcohol-related mortality among urban Lithuanian population aged 35-64 years during the past 20 years were determined. PMID- 15759453 TI - [24-h profile of arterial pressure in hypertensive patients working in rotatory teams in conditions of Far North (Tyumen Region)]. AB - AIM: To specify a 24-h profile of arterial pressure (AP) in hypertensive patients working in duty regime in the Far North (Tyumen Region). MATERIAL AND METHODS: AP parameters were studied in 155 males aged 25-59 with hypertension of stage I, II who were employed for duty work in the Far North areas and 38 control patients with hypertension stage I, II living in a moderate climatic zone (Tyumen). The groups were comparable by gender, age, duration of hypertension, office systolic and diastolic AP (SAP and DAP). All the patients have undergone 24-h monitoring of AP with assessment of basic mean parameters. RESULTS: The study group patients had scare symptoms and lower mean 24-h SAP, but high AP variability, high DAD as reflection of more significant structural changes of vessels and special functioning of the autonomic nervous system in the North. Mean 24-h AP showed more unfavourable changes in hypertensive subjects who had flight from Yamburg Moscow-Yamburg. CONCLUSION: The data of the study dictate the necessity to develop a differentiated risk strategy for health promotion, prevention and treatment of hypertension in those who work in the North of Tyumen Region in duty regime. PMID- 15759454 TI - [Clinicogenetic aspects of carbohydrate metabolism disorders and efficacy of their correction with moxonidine and metformine in patients with arterial hypertension]. AB - AIM: To study effects of monotherapy with moxonidine and metformine on metabolic parameters in hypertensive patients with carbohydrate dysbolism (CD) regarding polymorphic markers of genes PPARalpha, PPARgamma and IRS type 1 and 2. MATERIAL AND METHODS: A total of 83 patients (31 male and 52 female patients aged 40-75 years) with untreated arterial hypertension stage I, obesity and CD (by glucose tolerance test) entered the trial. The patients were randomized into two groups. Patients of group I (n=42) received moxonidin in a dose 0.4 mg/day, of group 2 (n=41)--metformin in a dose 1000 mg/day. Measurement of arterial pressure, blood count and biochemistry, oral test for glucose tolerance with glucose and insulin measurement before meal and 1, 2 and 3 hours later was made initially and on the treatment week 16 Genotypes of polymorphic markers of genes PPARA, PPARG2, IRS1 and IRS2 were defined in all the patients. RESULTS: Changes in basic hemodynamic and metabolic indices in therapy with moxonidine depending on polymorphic markers of genes PPARA, PPARG2, IRS1 and IRS2 in patients with AH and CD showed that G allele PPARG2 is associated with greater weight loss, G allele PPARA--with weight loss, C allele PPARA--with maximal fall of diastolic blood pressure. CONCLUSION: Genetic factors participate in development of metabolic disturbances in hypertensive patients, obesity and CD and determine treatment efficacy in each individual patient. PMID- 15759455 TI - [Effects of hyper- and dyslipidemias on activity of angiotensin-converting enzyme (biochemical disturbances in metabolic syndrome)]. AB - AIM: To study influence of lipoprotein complexes on activity of angiotensin converting enzyme (ACE). Material and methods. Lipoproteins fraction was isolated from the serum of 53 healthy males and females and was studied for lipoproteins action on ACE in vitro. In addition, the examination covered 783 individuals. The data were analysed mathematically for relationships between ACE activity and parameters of lipid metabolism. RESULTS: It was found that blood serum lipoproteins inhibit activity of soluble ACE. Serum ACE activity depends on concentration of total cholesterol (TC) and triglycerides. Maximal activity of ACE is observed in TC 9.41 mM/l and TG concentration 3.05 mM/l. Albumin-related free fatty acids affect ACE activity neither in aqueous solution nor in the presence of lipoprotein complexes. CONCLUSION: . Development of most common atherogenic hyperlipidemias raises 2-fold plasma ACE activity. Elevated ACE activity may be an integral component of biochemical disturbances in metabolic syndrome. PMID- 15759456 TI - [Differential diagnosis and treatment of vertigo in hypertensive patients]. AB - AIM: To study causes of vertigo in hypertensive patients and specify approaches to its treatment. Material and methods. Prevalence and causes of vertigo were analysed in 285 patients with arterial hypertension (AH). The examination included 24-h monitoring of arterial pressure (APM) and MR-tomography of the head. RESULTS: The majority of patients (78%) hospitalized with the diagnosis "hypertensive crisis" were diagnosed to have other diseases (headaches of tension, stroke, Meniere's syndrome and disease, etc.) the development of which was accompanied with hypertension and simulated a hypertensive crisis. Vertigo occurs in 20% hypertensive patients and is unrelated to elevated blood pressure. It is rather due to associated neurological, peripheral vestibular and other diseases. APM shows that vertigo occurs in hypotension after intake of hypotensive drugs. In hypertensive patients treatment of vertigo should not be directed only to management of elevated pressure but demands treatment of underlying disease. Vertigo plus mnestic disorders are effectively corrected with tanakan in a dose 120-160 mg/day. CONCLUSION: Vertigo in hypertensive patients is not caused by elevated pressure but related with concomitant neurological or peripheral vestibular diseases as well as hypotension. Hypertensive patients with vertigo need correction of the condition causing it. PMID- 15759457 TI - [Effects of vaccination against influenza on progression of chronic obstructive pulmonary diseases]. AB - AIM: To study antiinfluenza vaccination (AIV) effects on the course of chronic obstructive pulmonary disease (COPD) in metallurgists. MATERIAL AND METHODS: COPD and acute respiratory viral infections (ARVI) morbidity was studied in' metallurgists as well as a 5-year trend in functional tests in a group of 51 patients with moderate AR VI. RESULTS: It is shown that ARVI play a key role in activation of persisting infection in COPD. CONCLUSION: AIV reduces the rate and severity of COPD exacerbations, suppresses the disease progression. PMID- 15759458 TI - [Sicca syndrome in sarcoidosis and involvement of the salivary and lacrymal glands]. AB - AIM: To present differential-diagnostic signs of sarcoidosis with affection of the salivary and lacrymal glands and Sjogren's disease. MATERIAL AND METHODS: The examination of 620 patients with affection of the salivary and lacrymal glands revealed sarcoidosis in 19 of them. The diagnosis was verified histologically. Clinical, serological and histological characteristics of sarcoidosis patients were compared to those of 200 patients with Sjogren's disease (SD) detected among the examinees. RESULTS: Sarcoidosis patients vs those with SD (p < 0.001) had massive enlargement of the salivary glands (84.3%) with severe xerostomy which appeared rather early (78.9%), affection of the lacrymal glands manifesting with enlargement of the palpebral region, edema of the upper eyelids (57.9%), pulmonary lesion (78.9%), cranial nerves (47.4%), skin (42%), enlargement of the intrathoracic lymph nodes (100%). CONCLUSION: In spite of the presence of mucosal dryness simulating SD, sarcoidosis of the lacrymal and salivary glands has some specific features allowing differentiation of sarcoidosis. PMID- 15759459 TI - [Infectious complications in using mofetil micofenolate in patients with renal allotransplant]. AB - AIM: To study infectious complications in renal transplant recipients receiving mycophenolate mofetil (MMF) for prevention of acute transplant rejection or treatment of chronic allograft nephropathy (CAN). MATERIAL AND METHODS: A group of renal transplant recipients (n=47) receiving 1.0-2.0 g/day MMF with cyclosporine A (CsA) and steroids as maintaining immunosuppression was compared to a group (n=47) taking triple immunosuppressive therapy which included azathioprine (Aza). Separate group of patients (n=9) received MMF for treatment of CAN. In all groups etiology and incidence of infections were evaluated. RESULTS: During 2 years various posttransplant infections developed in 72.3% patients on MMF and 93.6% on Aza. The incidence of viral infections was 53.2% in MMF and 59.6% in Aza group, the incidence of bacterial infection--55.3 and 70.2%, respectively. Among 9 recipients with CAN the infections occurred in five. There were two cases of active tuberculosis in Aza group, one--in MMF group and one in patients with CAN. CONCLUSION: We suggest that MMF in the dose 1-2 g/day does not increase infection rates in renal transplant recipients comparing Aza. PMID- 15759460 TI - [Biliary tract in patients with chronic viral hepatitides]. AB - AIM: To characterize motor-kinetic and inflammatory changes in extrahepatic biliary tracts and gallbladder in patients with chronic viral hepatitis (CVH). To ascertain whether there is a pathogenetic correlation between affection of the biliary system and viral infection. MATERIAL AND METHODS: The condition of the biliary tract was examined in 183 patients with CVH using fractionated duodenal tubing with biochemical tests and bacteriological investigation of bile, dynamic ultrasonic investigation. RESULTS: 69.9% patients were diagnosed to have dysfunction of the Oddi's sphyncter caused by its hypertonicity, 4.4% patients had hypotonic sphyncter. Hyper- and hypotonicity of the gallbladder were observed in 45.8 and 20.8% patients, respectively. Biliary dysfunction was associated with the process activity but not with a nosological form of the disease. Chronic acalculous cholecystitis was verified in 21.2% patients. CONCLUSION: Chronic HBV and HCV infections are accompanied with biliary dysfunctions associated with activity of inflammation in the liver. Oddi's sphincter dysfunction in CVH is a risk factor of gallbladder inflammation. PMID- 15759461 TI - [Development of clinical thinking in medical students]. PMID- 15759462 TI - [Skin syndrome of drug disease]. PMID- 15759463 TI - [Nitric oxide: physiological and pathophysiological properties]. PMID- 15759464 TI - [G. A. Zakharyin--a founder of the Moscow Therapeutic School]. PMID- 15759465 TI - [Forum of leading specialists of sanatoria]. PMID- 15759466 TI - [Influence of heliometeofactors on sympathic and parasympathic activity as shown by analysis of cardiac rhythm variability in patients with ischemic heart disease and paroxysmal atrial fibrillation]. AB - Dynamics of heart rhythm variability (HRV) according to 24-h Holter monitoring of ECG was studied in patients with chronic IHD with paroxysmal cardiac fibrillation (PCF) and controls in response to unfavourable medical weather types and high geomagnetic activity. In unfavourable meteorological conditions IHD patients with PCF show attenuation of a general effect of autonomic nervous system on heart rhythm, changes in correlation of sympathetic and parasympathetic activity. Shifts to relative sympathicotony, parasympathicotony or absence of parasympathicotony or sympathicotony predominance were recorded. PMID- 15759467 TI - [Combined use of natural physical factors in patients with chronic cardiac insufficiency at a balneological resort]. PMID- 15759468 TI - [Thalassotherapy effect on cardiovascular system and cardiac rhythm variability]. AB - A complex study of physiological parameters allowed us to analyse reactions of cardiovascular system to dipping into a hot water varying in salt concentrations. These reactions differ from those to dipping into cold water. Dipping into hot water leads to depression of the tonicity of parasympathic nervous system in such a manner that increased heart rate does not cause vascular contraction. Effects of water salt concentrations on the above parameters need further investigations. PMID- 15759469 TI - [Rationale for treating patients with hypertension and atherosclerosis of the lower extremities with iodine-bromine baths containing molecular iodine]. AB - Higher therapeutic efficacy of iodine-bromine baths in conversion of iodine from an ionic form (standard iodine-bromine baths) into a molecular one is registered. This is explained by a 23.8 times increase in precipitation and a 4-times increase in iodine penetration into the body when it is molecular. A more effective function of the cardiovascular system, a stronger corrective effect on lipid metabolism, microcirculation in the absence of changes in a secretory function of the thyroid were registered. PMID- 15759470 TI - [Physiotherapy with rotating pulse magnetic field in combined therapy of chronic obstructive pulmonary disease]. AB - Rationale and technique are proposed and clinical trial has been made of efficacy of chronic obstructive pulmonary disease (COPD) treatment with impacts of rotating impulse magnetic field (RIMF) from the device Polyus-VIEM (two fields, induction 48-72 mTe, reversive rotation, 4-6 s reversion duration, 20-30 Hz, 7-10 min exposure of each field, 7-10 procedures). The addition of RIMF in combined treatment of COPD improves treatment and shortens its duration. PMID- 15759471 TI - [Use of external local low pressure to improve blood flow in children with infantile cerebral palsy]. AB - Cardiovascular system was studied in 16 children with spastic quadriplegic infantile cerebral palsy (ICP) aged 3-10 years. It was found at rheography that hemodynamics of ICP children differs from normal, in particular arterial tonicity and peripheral resistance were increased. The children were exposed to external local low pressure. This treatment reduced vascular tonicity, peripheral resistance and increased arterial elasticity. The effect is most noticeable on rheoencephalograms. PMID- 15759472 TI - [Ultrasound effects on lipid peroxidation in the liver and erythrocytes in experimental post-traumatic osteoarthrosis]. AB - We studied low frequency ultrasound effects on lipid peroxidation in the liver and erythrocytes of rabbits with experimental posttraumatic osteoarthrosis induced by transchondrial damage of the medial condyle of the right femoral bone (2 mm in diameter and 1 cm in depth in the direction of the medullar canal). Prior to treatment, rabbits with experimental osteoarthrosis had enhanced lipid peroxidation in the liver and erythrocytes. Ultrasound normalizes the above disorders in lipid peroxidation. PMID- 15759473 TI - [Efficacy of general magnetotherapy in conservative therapy of uterine myoma in women of reproductive age]. AB - Sixty women of the reproductive age with uterine myoma were divided into two groups. Thirty patients of the study group received combined therapy plus general magnetotherapy (GMT). Patients of the control group received only combined treatment. Ultrasound investigation registered a reduction in the size of myoma nodes by 16.7% in the study group, while in the controls myoma size did not change (p < 0.05). 1-year follow-up data for the study group demonstrated no cases of the myoma growth while 16.6% of the controls showed growth of myoma nodes, in 6.6% of them supravaginal myoma amputation was made for rapidly growing myoma. PMID- 15759474 TI - [Altered blood lipids spectrum in patients with stage I-II dyscirculatory encephalopathy and effects of treatment at a sanatorium]. PMID- 15759475 TI - [Ultraviolet radiation]. PMID- 15759476 TI - [Biochemical properties of mud deposits of some mineralized Siberian lakes]. PMID- 15759477 TI - [Ways of updating records for management of a sanatorium]. PMID- 15759478 TI - [Alexander Nikolaevich Ogilvi--the first director of the first balneological institute]. PMID- 15759479 TI - [Segmental-reflex massage in rehabilitation of patients with ischemic heart disease]. PMID- 15759480 TI - [Treatment and rehabilitation of pulmonological patients with use of climatic factors]. PMID- 15759481 TI - [Complex measures (yellow turpentine baths and interference currents) in the treatment of patients with diabetes mellitus with micro- and macroangiopathies]. PMID- 15759482 TI - [Characteristic features of the pathogenesis and treatment of coronary heart diseases, heart failure and arterial hypertension in patients with diabetes mellitus]. PMID- 15759483 TI - [Bronchial asthma and gastroesophageal reflux disease: are they interrelated by chance?]. PMID- 15759484 TI - [Clinical characterization and peculiarities of the clinical course of low activity hepatitis C]. PMID- 15759485 TI - [Blood levels of micro- and microelements in aged patients suffering from chronic coronal heart disease]. AB - The authors measured serum and whole blood levels of macro- and microelements (ME), such as iron, zinc, sulfur, potassium, calcium in 149 aged patients, suffering from coronary heart disease. The levels of the above ME were measured in various age groups: younger than 60, 60 to 74, 80 to 89 and older than 90 years old. In 36 aged women ME levels were established in correlation with bone mineral density. PMID- 15759486 TI - [Long-term application of warfarin or acenocumarol in patients with fibrillating arrhythmia: the effects compared]. AB - Thromboembolic complications (cerebral infarction and system embolism) are the most threatening ones in patients with ciliary arrhythmia without valvular lesions. Transoesofhageal echocardiography is the method of choice in detection of left atrial auricle thrombosis, which is the main source of thromboembolism in this category of patients. Primary and secondary prevention of thromboembolic complications in patients with ciliary arrhythmia is a topical problem, still remaining unsolved. Administration of indirect anticoagulants, which are the preparations of choice, demands strict doctor's supervision and continuous laboratory monitoring. The study presents authors' own data, based upon the observation of patients treated with either warfarin or acenocumarol. The paper demonstrates equal efficiency of both cumarine anticoagulants. The frequency of haemorrhagic complications after 12-month therapy with either warfarin or acenocumorol in patients with ciliary arrhythmia without valvular lesions was comparable. Warfarin provided more stable level of anticoagulation and thus long term warfarin therapy was characterized by lower risk of complication. PMID- 15759487 TI - [Functional features of acute non-Q-wave myocardial infarction and the influence of pre-hospital treatment with propranalol, heparin and aspirin on the prognosis of the disease]. AB - 193 patients with their first non-Q-wave myocardial infarction (NQMI) were examined with the aim of studying the functional features of NQMI and estimate the influence of pre-hospital treatment with propranalol (PP), heparin and aspirin. The study found that patients having NQMI with initial ST segment depression, unlike those with initial ST segment elevation, were characterized by diffuse coronary atherosclerosis, evident diastolic dysfunction and left ventricle (LV) remodeling, progressing with the course of time. Early administration of aspirin, heparin and PP reduced LV remodeling, improved diastolic function and prognosis in patients having NQMI with initial ST segment depression. PMID- 15759488 TI - [To the question of the possibility of oral insulin administration]. AB - The influence of oral insulin upon carbohydrate exchange in healthy volunteers was evaluated. One of the two forms of insulin used in the study was insulin, immobilized in a volume of polyacrylamide gel, modified by ovomucoid (proteolytic enzyme inhibitor), isolated from duck eggs; the other form was enteric-coated tablets bases on hydrogel substance. The oral administration of the hydrogel substance or the tablets resulted in the fall of blood glucose concentration by 34.9 +/- 4.4% and 28.6 +/- 6.9%, respectively. The maximal effect of the hydrogel substance and the tablets was observed at the 90th and the 180th minutes. PMID- 15759490 TI - [Morphofunctional changes in gastrointestinal tract in patients with psoriasis accompanied by chronic opisthorchosis]. AB - The paper covers clinical, morphological and functional aspects of gastrointestinal tract condition in patients with psoriasis accompanied by chronic opisthorchosis (CO). The authors examined 150 patients with psoriasis accompanied by CO, 100 patients having psoriasis without helminthiasis, 100 patients with CO and 30 healthy people. The gastric secretion was evaluated by means of the fractional test (both phases) with histamine stimulation; other diagnostic procedures included carbohydrate absorption evaluation (5-gram D Xylose absorption test) and Kamer test of fat absorption. The morphological condition of the gastric and intestinal mucosa was investigated by means of light and electron microscopy. The study revealed gastric secretory dysfunction and malabsorption in small and large intestines in patients with psoriasis and CO, clarified the relation between the duration of psoriasis and opisthorchosis and gastric secretory dysfunction and determined dependence of small intestine malabsorption on such factors as stage, severity, degree of skin involvement and duration of psoriasis. The authors also established interrelation between the above malfunctions and gastric and intestinal structural abnormalities. PMID- 15759489 TI - [The causes of death of patients with rheumatic diseases in Moscow]. AB - The study was held in order to analyze the main causes of death in cases of rheumatic diseases (RD) in Moscow. The authors studied the pathology records of autopsies performed in 1999-2002 in two pathology departments of Moscow clinics. Cases with RD were selected. The study found 165 cases of RD, which constituted 2% of all autopsies performed in these departments. There were 99 cases (60%) of rheumatic heart disease (RHD), 4 cases (2.4%) of rheumatic fever (RF) relapse, 28 cases (17%) of rheumatoid arthritis (RA), 8 cases (4.8%) of systemic lupus erythematosus (SLE), 3 cases (1.8%) of scleroderma systematica (SS), 2 cases (1.2%) of ankylosing spondylitis (AS), 2 cases (1.2%) of systemic vasculitis (SPV), 11 cases (7.3%) of osteoarthrosis, 3 cases (1.8%) of gout, 1 case (0.6%) of polymyositis. The death of patients with RHD had been caused by hemodynamic decompensation (HD) in 54% of the cases, acute cardiovascular collapse (ACC) in 14% of the cases, 6% of the patients had died from thromboembolism (TE) and 26%- from other conditions (intoxication, uremia, brain and lung edema etc). The death of patients with RF was caused by TE in 2 cases, by HD in 1 case and by ACC in 1 case. Secondary amyloidosis resulting in chronic renal failure and uremia occurred in 5 out of 28 cases of RA, HD--in 3, ACC--in 7, TE--in 1, infectious complications--in 5, other complications--in 7 cases. Patients with SLE died from various conditions: uremia in 2 cases, acute adrenal failure in 1 case, infectious complications in 2, ACC--in 2, brain edema--in 1 case. The complications of SS were uremia and intoxication. ACC was the cause of death in cases of gout and SS. The majority of RD cases were patients with RHD. The main cause of death in RD was cardiovascular disorders. PMID- 15759492 TI - [Hemodynamic and anti-ischemic effects of moexipril in patients having postinfarction heart dysfunction and moderate left ventricular heart failure]. AB - The purpose of the study was to evaluate hemodynamic and anti-ischemic effects of moexipril in patients with postinfarction left ventricular dysfunction (PL VD) and moderate left ventricular heart failure (NYHA functional class II-III). The subjects of the study were 21 male patients (aged 52.4 +/- 1.1 years) with CHD and PLVD, related to old large-focal anterior myocardial Q-wave infarction. 4 week treatment with ACF-inhibitor moexipril in an average dose of 10.7 +/- 0.4 mg per day clinically improved the patients' condition, which was characterized by 32.2% reduction of stenocardia attack frequency (p < 0.01) and 36.4% decrease in nitroglycerin demand (p < 0.01). Activity tolerance increased by 39.2% (p < 0.001), and echoCG found a 16.5% increase of LV ejection fraction (p < 0.05). Conjunctival biomicroscopy showed that moexipril substantially improved microcirculation (MC); total conjunctival index significantly decreased by 21.7%. The results of the 4-week moexipril administration demonstrate hemodynamic and anti-ischemic effects of the new ACF inhibitor, its positive influence on transcapillary oxygen exchange and MC in patients with PLVD, associated with NYHA functional class II-III heart failure. PMID- 15759491 TI - [Is anti-helicobacter therapy a rational approach in treatment of erosive and ulcerous lesions of the gastroduodenal mucosa in patients with inflamatory bowel diseases?]. AB - The study revealed changes in the oesophagogastroduodenal mucosa in 110 patients with IBD; in 60.9% of cases these changes were associated with Helicobacter pylori. 35 patients with IBD were examined to form two groups. The first group (20 patients) received rabeprazol (pariet) in a dose of 20 mg per day; the rest 15 patients were administered 120 mg of de-nol four times a day; amoxicicline and furazolidon were used as additional therapy in cases with Helicobacter pylori. The study showed that successful eradication did not always result in erosion epithelization but, on the contrary, only 40% cases of clinical and endoscopic remission were associated with Helicobacter pylori elimination. These data suggest that anti-helicobacter therapy is not a rational approach in treatment of this category of patients. PMID- 15759493 TI - [Chronic gastritis: last decade's achievements and problems]. AB - Classifications of chronic gastritis and neoplastic gastric diseases, developed in recent years (1996 Houston update of 1990 Sidney classification system, 2002 New Orlean classification of atrophic gastritis according to recommendations of International Group for Atrophy Studies; 1998 Padova classification of gastric displasia, and 1998 Vienna classification of gastrointestinal neoplasia) allow to statandardize international research and perform more objective diagnostics of pathological changes in the gastric mucosa. Studies carried out in recent years have established that morphological manifestations of chronic gastritis caused by Helicobacter pylori infection can be reduced after its eradication. Longterm treatment with proton pump inhibitors have been demonstrated not to cause atrophic changes in the gastric mucosa when undertaken after successful eradicational therapy. It has been established that corporal gastritis intensifies in patients treated with proton pump inhibitors. The studies show that measurement of serum levels of Helicobacter pylori antibodies, gastrine, pepsinogen I and II can be used in non-invasive serologic diagnostics of atrophic gastritis. Achievements in diagnostics and treatment of chronic gastritis create the necessary prerequisites for the development of gastric cancer preventing measures. PMID- 15759494 TI - [To the diagnostics of primary ciliary dyskinesia without situs inversus by the example of a clinical observation]. PMID- 15759496 TI - [Russian medical therminology of the 18th century]. PMID- 15759495 TI - [A multidisciplinary clinical discussion: a difficult patient with fever of unknown origin]. PMID- 15759497 TI - [A conformational study of kiotorphin and its biologically active analogue]. AB - The spatial and electronic structures of kiotorphin and its biologically active analogue [D-Arg2]-kiotorphin were studied. It was shown that [D-Arg2]-kiotorphin has a more rigid structure compared with the native molecule. The D-izomerization of arginine restricts the conformational mobility of the main chain of the molecule, which completely rules out its unfolded form. The electronic characteristics of the molecule in this case substantially change. PMID- 15759498 TI - [The interaction of hypochlorite with fatty acid hydroperoxides results in the generation of free radicals]. AB - The interaction of hypochlorite with linoleic acid hydroperoxides was studied by the coumarin C-525-enhanced chemiluminescence and ESR spin trapping techniques. Linoleic acid hydroperoxide was obtained in the reaction of lipoxygenase and linoleic acid. Alpha-(4-pyridyl-1-oxyl)-N-tert Butylnitron was used as a spin trap. It was shown that the addition of hypochlorite to the incubation media containing linoleic acid and lipoxygenase resulted in an intensive chemiluminescence flash. The intensity of this flash correlated with the hydroperoxide concentration. The analysis of ESR spectra of spin adducts produced in the reaction of hypochlorite with linoleic acid hydroperoxide showed the presence of O-centered, most likely peroxyl, radical with the splitting constants alphabetaH = 0.260 mT aN = 1.662 mT and C-centered penthyl radical with the splitting constants alphabetaH = 0.260 mT; aN = 1.662 mT. These data suggest that hypochlorite produced by phagocytes in vivo can induce the generation of free O- and C-centered radicals, promoters of free radical processes. PMID- 15759499 TI - [1H NMR analysis of the complex formation of aromatic molecules of antibiotic and vitamin in aqueous solution: heteroassociation of actinomycin D and flavin mononucleotide]. AB - The molecular mechanism of the combined action of antibiotic and vitamin was studied by NMR spectroscopy. The heteroassociation of the antitumor antibiotic actinomycin D and flavin mononucleotide was investigated as a function of concentration and temperature by 500 MHz 1H NMR spectroscopy. The equilibrium association constant, the thermodynamic parameters (deltaH, deltaS) of heteroassociation of actinomycin D with flavin mononucleotide, and the limiting values of proton chemical shifts in the heterocomplex were determined from the concentration and temperature dependences of proton chemical shifts of molecules. The most favorable structure of the 1:1 actinomycin D-flavin mononucleotide heteroassociation complex was determined using both the molecular mechanics methods (X-PLOR software) and the limiting values of proton chemical shifts of the molecules. In the calculated structure, the planes of the chromophores of actinomycin D and flavin mononucleotide molecules in the 1:1 heterocomplex are parallel and separated from each other by a distance of about 0.34 nm. At the same time, there is a probability of formation of intermolecular hydrogen bonds in the calculated structure of 1:1 actinomycin D-flavin mononucleotide complex. The analysis of the results obtained suggests that aromatic molecules of vitamins, e.g., flavin mononucleotide, can form energetically favorable heterocomplexes with aromatic antitumor antibiotics in aqueous solution, modulating thereby the efficacy of their medical and biological action. PMID- 15759500 TI - [Structural changes in Cry3A delta-endotoxin from Bacillus thuringiensis var. Tenebrionis in alcohol solutions at pH 2.0]. AB - Conformational changes in Cry3A delta-endotoxin caused by three different alcohols (ethanol, butanol, and isopropanol) were studied using the methods of circular dichroism, scanning microcalorimetry, and electron miscroscopy. It was shown that, in addition to the standard decrease in the native structure stability, the alcohols can cause a conformational transition that results in a sharp increase in the beta-structure content and a change in the environment of aromatic residues. The conformational transition is accompanied by intermolecular association, which leads to the appearance of oligomers in the form of short filaments. When the alcohols were removed, the oligomers dissociated again into monomers, but it is likely that the native structure either is not restored or is restored only in a small portion of molecules. The oligomer structure is rather cooperative, and its thermostability is higher than that of the initial structure. The disruption of this structure upon heating, observed as a heat absorption peak, is reversible. PMID- 15759501 TI - [The oxidation of sperm whale, horse, and pig oxymyoglobins, catalyzed by ferrocyanide ions: kinetics and mechanism]. AB - The influence of pH, ionic strength of the solution, and [Fe(CN)6]4- concentration on the rate of oxidation of sperm whale, horse, and pig oxymyoglobins, which is catalyzed by ferrocyanide ions, was studied. These myoglobins have homologous spatial structures and identical redox potentials but differ by the amount of His residues located on the protein surface. The effect of the MbO2 complexing with redox-inactive Zn2+ ion on the reaction rate was also examined. At the equimolar Zn2+ concentration, zinc ions form a stable complex with His119(GH1). It was found that the kinetic behavior of horse MbO2, which lacks His12(A10) substituted for by Gln, is fully analogous to one of sperm whale MbO2, while the oxidation of pig MbO2, three histidines of which, His12, His113(G14), and His116(G17), are replaced by Gln, is strongly inhibited. The mechanism of the catalysis was shown to involve specific binding of [Fe(CN)6]4- to the protein at the His119(GH1) site, which is in accord with the large positive electrostatic potential of this site and the presence here of a cavity large enough to accommodate [Fe(CN)6]4-. The nearby His113 and His116 residiues, which are absent in pig Mb, also play a very important role in the catalysis, because their protonation (especially of the last residue) is most likely responsible for the week oxidation of bound [Fe(CN)6]4- by dissolved oxygen. PMID- 15759502 TI - [Molecular reasons for the regulation of the enzymatic activity of bovine pancreatic deoxyribonuclease I as studied by dynamic light scattering and fluorescent spectroscopy]. AB - The effect of catalytic bivalent and inhibitory monovalent cations on the interactions of bovine pancreatic deoxyribonuclease I with the circle single stranded DNA of M13 phase was studied. It was found that monovalent cations affect the site of binding to DNA and the active center of the enzyme; in their presence, a break of the formation of the enzyme-substrate complex occurs. The results provide evidence that conformational changes of the protein molecule in all cases are more substantial that it could be expected from X-ray data published earlier. PMID- 15759503 TI - [Coherent singlet-triplet transitions at the initial step of tubulin assembly into microtubules]. AB - Quantum chemistry calculations [DFT-B3LYP QM/MM method, 6-31G** basis set, + ab initio molecular dynamics] were used to study the action of Mg2+ on tubulin properties. It was shown that the hydration of the guanosine triphosphate-tubulin forms a protein zone structure, which includes a electron-occupied zone and a conductivity zone. The binding of Mg2+ to guanosine triphosphate-tubulin results in the unpairing of electrons in the occupied zone (triplet state formation) followed by their transition to the conductivity zone in which the inversion of spin occurs (singlet state formation). The formation of triplet state is the initial step in the subsequent protein dynamics in the picosecond range of time. The dynamics shows up as a coherent oscillating transition of tubulin between the triplet and singlet states, which is evidence of a simultaneous adjustment between nuclear and electron configurations of the protein (ab initio molecular dynamics calculations). The barrier between the triplet and singlet states does not exceed 0.60 kcal x mol(-1). The barrier overcome is considered as electron tunneling through the Fermi surface, which separates the occupied and conductivity zones. Zone formation occurs in the presence of the shell of biological water surrounding the protein. PMID- 15759504 TI - [Polymerization of myorod, a surface protein of thick filaments of molluscan smooth muscle]. AB - The study is concerned with the polymerization of myorod, a protein from thick filaments of molluscan smooth muscles, which is an alternative product of the gene of myosin heavy chains. The dependences of the properties and polymer structure of myorod on the conditions of its formation were investigated. It was shown that myorod loses the ability to form viscous polymers after proteolytic removal of the unique sequence. It was supposed that the specificity of polymerization of myorod are determined by its unique N-terminal sequence. PMID- 15759505 TI - [P53 under low-intensity inluence of physical and chemical nature (ionizing radiation and antioxidant)]. AB - It was found that low-intensity ionizing radiation and the antioxidant fenozan at a low concentration (10(-14) M) produce opposite effects on the content of protein p53 in the blood serum of mice. Thus, low-intensity gamma-irradiation of AKR mice with a dose of 1.2 cGy (0.6 cGy per day) led to a decrease in the content of p53 and acceleration of leukosis, whereas fenozan, which has membranolytic and radioprotective properties, when injected intramuscularly to F1 mice (CBA + c57 black) increased the content of p53. However, the dynamics of the activation of protein p53 depended on the concentration of fenozan (10(-4) or 10( 14)), which may be due to the difference in its binding to the membrane and the changes in its antioxidative properties depending on concentration. PMID- 15759506 TI - [Stable isotopes of Mg2+ as activators of the suppressed ATP-generating function of mitochondria]. AB - The ATP-generating activity of rat myocardial mitochondria and intramitochondrial creatine kinase was examined as a function of the isotopy of the incubation medium magnesium pool. The study was performed using in vitro systems prepared from the hearts of animals injected with 1-methylnicotine amide, which suppresses the NAD (NADP)-dependent reactions in vivo. It was shown that the presence of the 25Mg paramagnetic cations essential by compensates for the intramitochondrial ATP deficiency caused by the 1-methyl-nicotine amide-induced blockade of oxidative phosphorylation. This effect is hardey achievable in systems where the magnesium pool consists of isotopes with a zero nuclear spin (24Mg, 26Mg). The restoration of mitochondrial ATP synthesis involves the participation of creatine kinase since the activity of the latter does not depend on 1-methyl-nicotine amide. In this case, the high efficiency of this restaration seems to be a spin-selective phenomenon which requires predominantly 25Mg2+ cations. A possible meaning of the data for further studies on the mechanisms of enzymatic catalysis regulation is discussed. PMID- 15759507 TI - [Photochemical activity, spectral properties, and structure of chloroplasts in leaves of Pisum sativum L. under iron deficit and root anaerobiosis]. AB - A combined effect of iron deficit and root anaerobiosis on the biochemical composition, functional activity, and structure of chloroplasts in pea leaves was studied. These factors are shown to affect the chlorophyll accumulation, causing leaf chlorosis. Iron deficit makes itself evident in the chlorosis of top leaves. In the case of root anaerobiosis, chlorosis damages lower plant layers. The destructive effects are summarized under the influence of both factors. The light harvesting complexes of photosystems are reduced to a greater degree under iron deficit; under root anaerobiosis, complexes of reaction centers of photosystem I and II are reduced. Nevertheless, even under the combined effect of these factors, all pigment-protein complexes and their functional activities are preserved in yellow leaves. The ultrastructure of chloroplasts is gradually reduced in the course of developing chlorosis. In the begging, intergranal sites of thylakoids are destroyed, which is typical for iron deficit, then granal sites are broken. However, even in yellow and almost white leaves, small thylakoids capable of forming stacking and small grana of 2-3 thylakoids are preserved. The destructive effects are summarized due to different mechanisms of action of iron deficit and root anaerobiosis on the structure and function of leaves under their combined effect. PMID- 15759508 TI - [Destruction of human blood cells upon interaction with detonation nanodiamonds in experiments in vitro]. AB - The influence of nanodiamonds synthesized by detonation on human red and white blood cells in vitro was studied. Nine nanodiamond samples produced by different manufacturers and two samples of modified nanodiamonds with high colloidal stability were used in the study. It was shown that nanoparticles cause the destruction of white cells and the hemolysis of erythrocytes. Possible mechanisms of the desctructive effect of detonation nanodiamonds on blood-forming elements are discussed. PMID- 15759510 TI - [The activity of natural killer cells in healthy and tumor-bearing mice after treatment with low-intensity laser light]. AB - The effect of low-intensity laser light on the activity of natural killer cells from healthy and tumor-bearing mice was studied. Skin in the zone of the thymus or hind limb was illuminated, the remaining body surface being screened. The illumination was carried out for 30 days, with the duration of a single exposure being 1 min and intervals between the exposures being 48 h. The effect of laser light depended on the location of the illuminated area. It was shown that the exposure of the thymus of healthy animals for 20 and 30 days leads to a significant decrease in the activity of natural killer cells. On the contrary, the illumination of the limb for 10 or 20 days increased the activity of natural killer cells; but when hind limbs were treated for 30 days, the activity of natural killer cells decreased. Whereas tumor growth increased the natural killer cell activity, the illumination of tumor-bearing mice lowered the adaptive antitumoral resistance by decreasing the activity of natural killer cells. PMID- 15759509 TI - [Structural modifications of the surface of Escherichia coli bacteria and copper induced permeability of plasma membrane]. AB - The effect of Cu2+ on the structural organization of the cell surface of Escherichia coli bacteria during the induction of conductivity of a plasma membrane was studied. A fluorescent study did not reveal any substantial changes in the microviscosity of lipids by the action of copper ions. At the same time, a substantial reorganization of membrane proteins during plasmolysis was observed. A model of the copper-induced structural reorganization of membrane lipids was constructed, according to which the reorganization leads to the opening in the membrane of channels of nonspecific conductivity for cations. The opening of conductivity channels results from the break of disulfide bonds in critical membrane proteins during the interaction with Cu+, which form either due to the reduction of Cu2+ on specific sites of cell surface or by means of external reducing agents. PMID- 15759511 TI - [The functional characteristics of electroreceptive central neurons of the sea catfish Plotosus anguillaris]. AB - With the use of microelectrode techniques (extracellular recordings) and the method of post-stimulus histograms, the functional characteristics of medulla oblongata neurons of sea catfish Plotosus were investigated under stimulation of electroreceptors by a homogeneous electric field of different duration, intensity, and direction. Two types of the cells possessing, accordingly, tonic or phase activity were registered among 66 neurons investigated. The mode of responses (inhibition or acceleration) of tonic neurons to the direction of the applied electric current is typical for central neurons of fresh-water catfish connected with ampullae's electroreceptors. Neurons showing a substantial response to fields of an intensity less than 1 microV/cm were registered. The reactions were most pronounced with the duration of electric stimuli in the range of 20-200 ms; however, particularly sensitive neurons showed distinct responses to stimuli of duration of 5 and even 2 ms. Thus, for the first time a high sensitivity of ampullae's electroreceptors to high-frequency stimulus was discovered, which allows one to expand the range of studying electric signals used by weakly electric fish for electrolocation and communication. PMID- 15759512 TI - [The rotation of autowaves as a result of their penetration through a system of unexcitable obstacles. A mechanism of arrhythmias associated with aging]. AB - The dependence of the frequency of occurrence of excitation vortices rotating around unexcitable obstacles on the size and the number of the obstacles and also on the medium excitation threshold was studied. It was shown that the vortex formation takes place in a wide range of the model parameter values. The assumption was formulated that the mechanism of formation of excitation vortices under study underlies the increase in the heart arrhythmias associated with aging. PMID- 15759513 TI - [Thermal effects of a short light pulse on biological tissues. II. Light and heat fields]. AB - A set of heat conduction equations for a two-component medium simulating biological tissues were formulated. Their solutions were obtained, and the spatial distribution of light and temperature over tissue depth at different times after irradiation by a short light pulse was studied. The local absorption of light by blood vessels and the influence of this effect on the optical parameters of the medium, a more intense heating of blood as compared with its surrounding tissue, heat exchange between them, and heat transfer at the interface with different environments were accounted for. The solutions are expressed via characteristic times of the respective thermal processes to enable one to easily and vividly analyze the features of tissue heating and the influence of optical and thermal parameters on the temperature distributions of the components. The calculations are illustrated by examples. PMID- 15759514 TI - [A comparative analysis of restoration of electroencephalographic and protein synthesizing activities in neocortex and hippocampus in hibernating (ground squirrels) and nonhibernating (rats) animals during exit from hypothermia]. AB - A similarity in the sequence of restoration of the EEG spectrum between ground squirrels arousing from torpor and rats passing out of artificial hypothermia (17 18 degrees C) was shown. First of all, the low-frequency part of the EEG spectrum was restored. As animals warmed up, their breathing became hurried, cold shivering appeared, and the theta- and alpha-rhythms increased. During the exit from hypothermia, the activity of the protein-synthesizing system in both rats and ground squirrels was almost entirely restored when the animal body temperature achieved 21-22 degrees C. In ground squirrel, the rate of protein synthesis in the neocortex was lower than in hippocampus CA1 and CA3 areas, whereas in rats, on the contrary, it was higher in the neocortex in comparison with the CA3 area. PMID- 15759515 TI - [What is the subject of science "bioinformatics"?]. AB - The paper is concerned with some problems of terminology, in particular the term "bioinformatics". In the last few years, the term "bioinformatics" has been intensively used among molecular biologists to indicate a subject that is only a constituent of genomics and is considered to involve a computer-assisted analysis of all data on nucleotide sequences of DNA. However, a wide circle of scientists, including biologists, physicists, mathematicians, and specialists in the field of cybernetics, informatics, and other disciplines have accepted and accept, as a rule, the "bioinformatics" as a synonym of science cybernetics and as a successor of this science. In this case, the subject of science "bioinformatics" should embrace not only genomics but practically all sections of the biological science. It should involve a study of information processes (storage, transfer, and processing of information, etc.) participating in the regulation and control at all levels of living systems, from macromolecules to the brain of higher animals and human. PMID- 15759516 TI - How many students turn their backs on nursing? PMID- 15759517 TI - The e-record revolution. PMID- 15759519 TI - Otitis media. PMID- 15759518 TI - Raising awareness of the issues surrounding heart failure. AB - This article highlights some of the key elements of care required to provide equity for heart failure sufferers and to raise awareness of the work of the British Heart Foundation (BHF), which funds and supports specialist heart nurses around the UK caring for patients with heart conditions including heart failure. The BHF is running a Real Valentine appeal to raise 1m pounds sterling to fund 30 more specialist heart nurses. PMID- 15759520 TI - Domperidone. PMID- 15759521 TI - Improving physical health care in a mental health trust. AB - It is a challenge both to general medicine and psychiatry to develop systems that are better equipped to meet the physical health needs of mental health patients and reduce mortality rates (Hansen et al, 1997). Central and North West London Mental Health NHS Trust is establishing a department of physical health care staffed by nurses with general nursing backgrounds to tackle this problem. This nurse-led department is designed to train nurses and doctors in key aspects of physical health assessment and treatment interventions. PMID- 15759522 TI - How contraception nurses can improve teenage sexual health. AB - The rising incidence of unwanted pregnancies and sexually transmitted infections among teenagers has made teenage sexual health a government priority. Contraceptive nurses can play a key role in advising young people about sexual health issues and contraception. This article discusses the factors affecting teenage sexual activity and how nurses can help young people take responsibility for their sexuality. PMID- 15759523 TI - Understanding pulse oximetry helps effective assessment. PMID- 15759525 TI - How to carry out a clinical audit. PMID- 15759524 TI - Living with severe hypoxic COPD: the patients' experience. AB - AIM: To describe the experience of living with severe oxygen-dependent chronic obstructive pulmonary disease (COPD). METHOD: Unstructured, tape-recorded interviews were used to collect data from 10 patients with COPD. The interviews were undertaken in the patients' own homes. RESULTS: Recurrent themes emerged from the data. All participants reported physical and emotional problems, mainly caused by breathlessness. Half of the sample discussed feeling depressed and 80 per cent discussed how COPD had affected their family life. Participants discussed how difficult it was to stop smoking, and the guilt they felt when they realised their smoking had caused their disease. Half of the sample discussed needs, such as better communication between health professionals and patients about the diagnosis and prognosis. Long-term oxygen therapy was described in negative terms by 10 per cent of the sample. CONCLUSION: Severe COPD has an impact on all aspects of a patient's quality of life. A palliative, interdisciplinary approach from primary and secondary care, social services and the voluntary agencies needs to be adopted at an earlier stage of the disease. Patients should be encouraged to participate in the planning and management of their treatment to improve their experience of the disease in the future. PMID- 15759526 TI - A randomized controlled trial of botulinum toxin on lower limb spasticity following acute acquired severe brain injury. AB - OBJECTIVE: To determine whether serial casting combined with botulinum toxin reduces the development of calf contracture after severe head injury. DESIGN: A double-blind placebo-controlled trial of three parallel treatments for lower limb spasticity. SETTING: Acute general hospital in the UK. SUBJECTS: Adults aged 17 70 years admitted to hospital following a severe brain INTERVENTIONS: Current physical treatment (group I), lower leg casting plus injections with either saline (group II), or with botulinum toxin (group III) into gastrocnemius and soleus muscles. MEASURES: Limit of ankle dorsiflexion at entry and exit after up to 12 weeks, the Glasgow Outcome Scale (GOS) and Modified Ashworth Scale (MAS). RESULTS: Two hundred and fifty-three patients were screened and 35 were entered into the trial. Three patients died and four failed to complete the trial. Eighty eight per cent of those entering the randomized part of the study developed spasticity within 14 days of their injury and the mean range of improvement in the angle of passive ankle dorsiflexion was 4.59 degrees in controls, 11.69 degrees in cast and saline and 13.59 degrees in cast and botulinum toxin. There were significant improvements in the MAS scores in actively treated groups, but not in controls. Cast and botulinum toxin patients also demonstrated a significant improvement in the GOS. CONCLUSIONS: Active intervention with casting prevents talipes equinovarus deformities in patients losing ankle movement following severe brain injury. Casting alone in these patients is sufficient; the role of additional botulinum toxin needs further investigation, but is safe in these patients. PMID- 15759528 TI - Effect of recommended positioning on stroke outcome at six months: a randomized controlled trial. AB - OBJECTIVE: To evaluate the effect on patient outcome of a teaching package for nurses designed to improve the positioning of stroke patients. DESIGN: Cluster randomized controlled trial with six-month follow-up. SETTING: Ten stroke rehabilitation hospital units located within one UK inner city region. These were randomized to control or intervention group. SUBJECTS: A sample of 120 patients admitted within four weeks of a first stroke and with a hemiplegia. No eligible patient refused to participate. Eighty-three (69%) completed the study. INTERVENTION: All nursing staff on the intervention units received a group teaching package to improve their clinical practice in patient positioning. MAIN OUTCOME MEASURE: Rivermead Mobility Index (RMI) at six months post stroke. Patient's position was recorded using an established observational tool. RESULTS: After the teaching there was some evidence of better positioning in the intervention than the control group (difference in percentage of correct positions per patient 4.9%, 95% confidence interval (Cl -0.1% to 9.9%, p = 0.055). There was no evidence of differences between the two groups in any of the outcome measures at six months although there was a trend towards increased elbow flexor tone in the control group. CONCLUSIONS: A teaching intervention to improve patient positioning made no significant impact on outcome at six months post stroke. However, following the teaching there was only a slightly higher incidence of recommended patient positioning within the intervention group. Thus, a teaching package may not be powerful enough to enable any effect on patient outcome to be measured. PMID- 15759527 TI - A study to assess the effect of nursing interventions at the weekend for people with stroke. AB - OBJECTIVE: To examine whether additional therapy provided by nurses at the weekend improved the physical outcome for people with stroke on a stroke rehabilitation unit. DESIGN: A single blind randomized controlled trial. SETTING: A 16-bed stroke rehabilitation unit in the north of England. SUBJECTS: Forty-one people with stroke were randomized by means of minimization to intervention and control groups. INTERVENTIONS: The intervention group received additional exercise at the weekend provided by the nursing staff and the control group received their usual care. Both groups received usual care during weekdays. MAIN OUTCOME MEASURES: The Motor Assessment Scale (MAS), the Barthel Index (BI) and length of stay in hospital. RESULTS: No significant differences were found between the groups in terms of MAS and BI at discharge but there was a borderline significant difference between the groups on unconditional testing in terms of length of stay in hospital and on the stroke unit (p = 0.05 and p = 0.07 respectively). However, these findings were in favour of the control group. On conditional testing (adjusting for BI on admission and age) these differences disappeared (p = 0.14 and p = 0.15) for length of stay in hospital and on the stroke unit respectively. CONCLUSIONS: The present study indicates that an increase in one-to-one input by nurses for people with stroke did not lead to a measurable difference in outcome in this small study. PMID- 15759529 TI - Comparison of the impact of a single session of isokinetic or isotonic muscle stretch on gait in patients with spastic hemiparesis. AB - OBJECTIVE: To establish if isokinetic and isotonic muscle stretch (with or without weight-bearing) of the ankle plantar flexors improves gait in hemiplegic patients. A further aim was to compare the effectiveness of these treatment methods. DESIGN: A randomized, parallel group prospective study. SETTING: A stroke rehabilitation unit. SUBJECTS: Ambulatory hemiparetic stroke patients with mild to moderately severe muscle hypertonia of the lower limb and a group of healthy control subjects. INTERVENTION: Subjects were randomized to receive a single 20-min session of isokinetic muscle stretch or isotonic muscle stretch with or without weight-bearing. OUTCOME MEASURES: Selected kinematic, kinetic and spatio-temporal gait parameters were measured at baseline, immediately after the muscle stretch and 24 h later. RESULTS: Sixty-six patients and 21 healthy control subjects were recruited and completed the study. There were statistically significant differences between the patient groups and the healthy subjects on most of the gait parameters studied. However, the differences between the patient groups or between the three measurements over time for each type of muscle stretch did not reach statistical significance. CONCLUSIONS: A single session of isokinetic or isotonic muscle stretch (with or without weight-bearing) of the ankle plantar flexors has no clinically observable effect on the gait of hemiplegic stroke patients. PMID- 15759530 TI - Efficacy of Bobath versus orthopaedic approach on impairment and function at different motor recovery stages after stroke: a randomized controlled study. AB - OBJECTIVE: To investigate the effectiveness of Bobath on stroke patients at different motor stages by comparing their treatment with orthopaedic treatment. DESIGN: A single-blind study, with random assignment to Bobath or orthopaedic group. SETTING: Physical therapy department of a medical centre. SUBJECTS: Twenty one patients with stroke with spasticity and 23 patients with stroke at relative recovery stages participated. INTERVENTIONS: Twenty sessions of Bobath programme or orthopaedic treatment programme given in four weeks. MAIN OUTCOME MEASURES: Stroke Impairment Assessment Set (SIAS), Motor Assessment Scale (MAS), Berg Balance Scale (BBS) and Stroke Impact Scale (SIS) for impairment and functional limitation level. RESULTS: Participants with spasticity showed greater improvement in tone control (change score: 1.20 +/- 1.03 versus 0.08 +/- 0.67, p = 0.006), MAS (change score: 7.64 +/- 4.03 versus 4.00 +/- 1.95, p = 0.011), and SIS (change score: 7.30 +/- 6.24 versus 1.25 +/- 5.33, p = 0.023) after 20 sessions of Bobath treatment than with orthopaedic treatment. Participants with relative recovery receiving Bobath treatment showed greater improvement in MAS (change score: 6.14 +/- 5.55 versus 2.77 +/- 9.89, p = 0.007), BBS (change score: 19.18 +/- 15.94 versus 6.85 +/- 5.23, p = 0.015), and SIS scores (change score: 8.50 +/- 3.41 versus 3.62 +/- 4.07, p = 0.006) than those with orthopaedic treatment. CONCLUSION: Bobath or orthopaedic treatment paired with spontaneous recovery resulted in improvements in impairment and functional levels for patient with stroke. Patients benefit more from the Bobath treatment in MAS and SIS scores than from the orthopaedic treatment programme regardless of their motor recovery stages. PMID- 15759531 TI - A pilot study of the effect of aerobic exercise on people with moderate disability multiple sclerosis. AB - OBJECTIVE: A pilot study to investigate the effect of aerobic exercise on the mobility and function of people with moderate disability multiple sclerosis (MS). DESIGN: A small group, comparative, pre- and post-intervention study. SETTING: A gymnasium within a general hospital. SUBJECTS: Eight subjects, all female, average age 45 years (range 33-61) with moderate disability MS (Kurtzke scale 4 6), were recruited; six completed the study. INTERVENTION: Intervention consisted of bi-weekly sessions of 30 min cycling on a static bike at their maximal level of exertion for 12 weeks. MAIN MEASURES: The stability of each individual's condition was established pre-intervention using three baseline assessments over two weeks. A battery of tests (the 10-metre and 6-min walk tests, the Functional Reach, the Gulick and the Guys Neurological Disability Scale) were completed pre and post intervention. Potential negative effects were recorded on the Fatigue Severity Scale and the Modified Ashworth Scale. An independent rater completed assessments post intervention. Inter-rater reliability was found to be acceptable. RESULTS: Comparisons pre and post intervention (Wilcoxon signed ranked test) showed significant improvement on the Guys Neurological Disability Scale (p = 0.026), with the mean score reducing from 13 to 9, and the 6-min walk test (p = 0.046), with the mean distance increasing from 200 m to 261 m. Other measures failed to reach significance. CONCLUSIONS: Findings from this pilot study suggest overall disability and mobility improved with the aerobic training. A positive treatment effect in this small study suggests the need for a larger trial. Knowledge of how to establish predictive heart rate and how to monitor the effects of cycling with people with MS will be used to inform future clinical trials. PMID- 15759532 TI - Is it possible to actively and purposely make use of plasticity and adaptability in the neurorehabilitation treatment of multiple sclerosis patients? A pilot project. AB - OBJECTIVE: To investigate whether neurorehabilitation is able to influence clinical parameters and brain function measured radiologically. DESIGN: A group of healthy probands was compared with two groups of multiple sclerosis (MS) patients, one of which received rehabilitative therapy. SETTING: Outpatient in a university hospital. SUBJECTS: Twenty-eight patients with multiple sclerosis (MS), 17 of whom received rehabilitative therapy, and 13 healthy controls. INTERVENTIONS: Two months of rehabilitative eclectic therapy based on principles of sensorimotor learning and adaptation. MAIN MEASURES: Multiple Sclerosis Functional Composite, Modified Fatigue Impact Scale, Beck Depression Inventory Score, Barthel Index, Environment Status Scale and Multiple Sclerosis Quality of Life--54, and functional magnetic resonance imaging (fMRI). RESULTS: Patients who underwent neurorehabilitation showed a greater drop in fatigue, depression, impairment, disability and handicap and more improvement in quality of life than those who did not receive therapy. Correlation of brain activity between the right and the left hemisphere is greater in healthy individuals than in MS patients. Neurorehabilitation resulted in a trend for increased correlation between the left and the right hemisphere in patients (approaching the standard). In comparison with control groups, signal amplitudes in anatomical areas did not show any significant changes. CONCLUSION: Clinical changes seen with neurorehabilitation were not associated with any detectable changes in fMRI observations. PMID- 15759533 TI - Measuring energy expenditure using heart rate to assess the effects of wheelchair tyre pressure. AB - OBJECTIVE: To investigate the effects of wheelchair tyre pressure on mechanics and energy and explore the use of heart rate as a measurement of energy expenditure. DESIGN: A single factor repeated measures design was used. Four tyre pressures (100, 75, 50, 25 psi) represented a change of workload. Each subject wheeled at a constant self-selected wheeling velocity for 8 min. A total of four trials were completed with a 10-min rest between trials. Oxygen consumption, heart rate and distance travelled were collected during each trial. SUBJECTS: Three women and 11 men with spinal cord injury. The mean age for the whole group was 34.5 years. The range of lesion level was T4-L1. RESULTS: There was a significant increase is energy expenditure when tyres were deflated to 50 psi from 100 psi. The mean correlation between heart rate and oxygen consumption was 0.74 for all subjects. For the subjects with lesions above T6 and T6 and below the correlations were 0.55 and 0.82, respectively. CONCLUSIONS: Tyre pressures below 50% inflation add an additional 25% increase in energy expenditure during wheeling. This could be detected using oxygen consumption or heart rate, as heart rate was shown to have a good correlation with oxygen consumption in the spinal cord injured with lesions below T5. Heart rate does have its limitations and it should only be used to measure within-subject differences. PMID- 15759534 TI - Sustained attention in patients with mild traumatic brain injury. AB - OBJECTIVE: To demonstrate that two tests of sustained attention were sensitive to attention deficits in patients with mild traumatic brain injury (TBI). DESIGN: A cross-sectional study recruiting 51 patients with TBI and 51 matched controls. OUTCOME MEASURES: The Sustained Attention to Response Task (SART) and Monotone Counting Test. RESULTS: The patient groups performed significantly worse than the normal controls in both sustained attention tests. The SART performance was also correlated with pathology severity in the patient group in terms of loss of consciousness (r = 0.247, p = 0.05). A cut-off of less than 1 standard deviation (SD) gives optimal diagnostic information in terms of sensitivity in the present sample (0.61 for Monotone Counting Test; 0.75 for SART). CONCLUSION: These findings suggest that the SART and Monotone Counting Test are sensitive to patients with mild TBI. The SART-assessed sustained attention is also sensitive enough to detect attention impairment in this clinical group regardless of diagnosis and may provide clinicians with an alternative method of assessing sustained attention in these clinical groups. PMID- 15759535 TI - Improving patient and carer communication, multidisciplinary team working and goal-setting in stroke rehabilitation. AB - OBJECTIVE: To determine the extent to which three forms of multidisciplinary team (MDT) care in stroke rehabilitation meet the standards set by the United Kingdom National Service Framework (NSF). DESIGN: Consecutive assessment of the three forms of care was completed. SUBJECTS: The study included three groups of 25 stroke inpatients on the stroke rehabilitation ward. INTERVENTION: (1) A standard weekly MDT meeting using a standard form for documentation; (2) a standard MDT meeting using a newly devised form; and (3) a novel MDT ward round using the new form, and attended by doctors. RESULTS: MDT ward rounds result in significantly better consideration of patients' needs (median 7 per patient compared with 0 and 5 in phases one and two), enhanced SMART (specific, measurable, achievable, realistic and time framed) goal-setting (median 3 per patient compared to 1 in phases one and two); greater patient involvement (12 patients compared to 0 and 4 in phases one and two); and improved team working (measured using the team climate inventory) than do MDT meetings. CONCLUSIONS: In the present study, standard weekly MDT meetings did not meet the standards set for MDT care by the NSF. The use of a MDT ward round allows these standards to be achieved. PMID- 15759536 TI - Vocational perspectives after spinal cord injury. AB - OBJECTIVE: To give insight into the vocational situation several years after a traumatic spinal cord injury (SCI) and describe the personal experiences and unmet needs; to give an overview of health and functional status per type of SCI and their relationship with employment status. DESIGN: Descriptive analysis of data from a questionnaire. SETTING: Dutch rehabilitation centre with special department for patients with spinal cord injuries. SUBJECTS: Fifty-seven patients with a traumatic SCI, aged 18-60 years, admitted to the rehabilitation centre from 1990 to 1998. MAIN MEASURES: Questionnaire with items related to vocational outcome, job experiences, health and functional status. RESULTS: Of 49 patients who were working at the moment of SCI 60% currently had a paid job. Vocational outcome was related to a higher educational level. A significant relation between the SCI-specific health and functional status and employment was not found. The respondents who changed to a new employer needed more time to resume work, but seemed more satisfied with the job and lost fewer working hours than those who resumed work with the same employer. In spite of reasonable to good satisfaction with the current work situation, several negative experiences and unmet needs were reported. CONCLUSIONS: Despite a high participation in paid work following SCI, the effort of the disabled worker to have and keep a job should not be underestimated. PMID- 15759537 TI - Evaluation of traumatic brain injured patients in correlation with functional status by localized 1H-MR spectroscopy. AB - OBJECTIVE: To determine whether proton magnetic resonance spectroscopy (1H-MRS) could be a useful tool for detecting microscopic diffuse axonal injury to evaluate the functional status of patients with traumatic brain injury (TBI). DESIGN: A comparative study. SETTING: An inpatient rehabilitation unit in Korea. SUBJECTS: We examined eight adult patients who had severe TBI approximately five months before. Fourteen normal controls were employed for comparison. MAIN MEASURES: Image-guided localized in vivo 1H-MRS was performed in the parietal white matter and occipital grey matter regions in brain in which no definite abnormalities in MR imaging were found at the time of 1H-MRS examination. We evaluated functional status for all patients with the Functional Independence Measure (FIM) on the same day as the 1H-MRS examination and compared the results. RESULTS: In the parietal white matter, the [N-acetyl aspartate/creatine] ratios were significantly lower, and the [choline/creatine] and [myo-inositol/creatine] ratios were significantly higher than those of normal controls. Significant correlations of the IN-acetyl aspartate/creatine] and [myo-inositol/creatine] ratios in the parietal white matter with FIM scores were observed (p < 0.05). CONCLUSIONS: A spectral feature of the decreased [N-acetyl aspartate/creatine] and increased [choline/creatine] and [myo-inositol/creatine] ratios in parietal white matter may be a marker for diffuse axonal injury in patients with TBI and has a significant correlation with the functional status of the patients. Localized 1H-MRS has the potential to be used for detecting diffuse axonal injury in vivo in TBI patients, which can be used to guide evaluation of the functional status of TBI patients receiving rehabilitation. PMID- 15759538 TI - The reliability of balance, mobility and self-care measures in a population of adults with a learning disability known to a physiotherapy service. AB - OBJECTIVE: To estimate the reliability of three measures of balance, mobility and activity for use in clinical and research physiotherapy, with adults with a learning disability. DESIGN: Prospective study to investigate test-retest and inter-rater reliability. SETTING: Participants' homes and day centres. MEASURES: The Berg Balance Scale (BBS), the Rivermead Mobility Index (RMI) and the Barthel Activities of Daily Living Index (BI). PARTICIPANTS: Of the 181 adults known to the Nottingham Community Physiotherapy Service for Adults with Learning Disabilities, 64 with a known Rivermead Mobility score of less than three were excluded. Of 117 randomized, a further 21 were found to fail this criteria, 27 had acute medical, social or behavioural problems, 22 were unable to participate or refused: therefore 47 entered the study. METHODS: Participants were visited in their own homes by two researchers on two occasions, one week apart and rated independently by each rater. Agreement was assessed with the kappa statistic (kappa) and percentage agreement for each item in each scale, and described using standard classification. Intraclass correlation coefficients for inter-rater and test-retest total scores and average differences of total scores, their standard deviations and limits of agreement, were calculated. RESULTS: For inter-rater observations, the Barthel Index and the Rivermead Mobility Index had almost perfect agreement (kappa = 0.86-1.00 and 0.89-1.00 respectively), with the Berg Balance Scale having substantial to almost perfect agreement (kappa = 0.74-1.00). For test-retest comparisons, both the Barthel Index and the Rivermead Mobility Index demonstrated moderate to almost perfect agreement (kappa = 0.57-1.00 and 0.45-1.00 respectively). Kappa scores for the Berg Balance Scale varied from low to almost perfect agreement (kappa = 0.37-1.00). CONCLUSIONS: The Berg Balance Scale, Rivermead Mobility Index and Barthel Activities of Daily Living Index are all reliable clinical and research tools for physiotherapists working with adults with learning disabilities. PMID- 15759539 TI - An investigation of the impact of the Force Sensing Array pressure mapping system on the clinical judgement of occupational therapists. AB - OBJECTIVES: To examine the impact of pressure mapping technology on the clinical decisions of occupational therapists and to examine the role of the Braden Scale in assisting with the selection of pressure-reducing cushions. DESIGN: Case studies. SETTING: Community. SUBJECTS: Forty clients. INTERVENTIONS: Clients were pressure mapped on their current seating surface and on four pre-selected cushions by the principal researcher. An occupational therapist completed the Braden Scale and a decision tree to assist in recommending a suitable pressure reducing cushion. MAIN OUTCOME MEASURES: Interface pressure maps, Braden Scale, and the cushion recommended, using a decision tree to guide clinical judgement. RESULTS: Thirty per cent (12) of the 40 cushions recommended were changed when the pressure maps from the Force Sensing Array (FSA) system were viewed. In 70% (26) of cases, the maps supported the cushion recommended. In 25% (10) of the cases, the maps showed that the client's current seating surface was unsuitable. After viewing the pressure maps, a surface other than the client's current surface was recommended in 47% (19) of the cases. There was a lack of agreement between the risk level of the clients as identified by the Braden Scale score, and the risk level of the clients as identified by the occupational therapist using a decision tree and the FSA maps. CONCLUSION: Pressure mapping technology has a positive impact on clinical decisions regarding the provision of pressure reducing cushions. Future research should examine the predictive validity of this technology. The Braden Scale may underpredict the risk level of the clients. PMID- 15759540 TI - The role of calcium and calcium channel blocking drugs in damage to the liver preserved for transplantation. AB - Calcium plays a crucial role in physiological process in liver cells however in high concentrations these ions can be pathogenic and lead to cell death. Mechanisms responsible for maintaining calcium ion concentration gradient in physiological conditions include transmembrane transport, storage in intracellular organelles and binding to cytoplasmatic proteins. Ischemia, sepsis, anoxia and action of toxins are responsible for uncontrolled influx of calcium and consequently cell damage. Liver damage during its preservation for transplantation is connected with dysfunction of many enzymes, damage of cell membrane and cytoskeleton proteins. During reperfusion Kupffer cells are activated, reactive oxygen species are produced and microcirculation is disordered by calcium--dependent processes. Calcium channel blocking (CCB) drugs exhibit immunomodulatory impact and positive interaction with cyclosporine or tacrolismus. They also have cytoprotective properties during preservation end reperfusion time. They seem to improve liver function, decrease liver cell damage, elevate bile production, decrease lipid peroxydation and free radicals production. But in some experiments CCB do not modify calcium concentration. More research on preservation conditions is needed to increase the probability of a successful liver transplantation. PMID- 15759541 TI - Costimulatory pathways as a basic mechanisms of activating a tolerance signal in T cells. AB - Immune tolerance is an active response leading to the T cell unresponsiveness in the presence of the graft, which may develop through a couple of mechanisms including costimulation blockade. The CD28/B7 and CD40L/CD40 costimulatory pathways have been described as the critical for T cell activation. When activated T cell upregulate CTLA4, which importance as a negative regulatory costimulatory molecule is highlighted by the recent evidence suggesting that CTLA4 may function as a master switch for peripheral T cell tolerance. The effects of CTLA4 engagement are directed at the inhibition of CD28 signaling. Modulation of proximal TCR signals and down-stream effector pathways of T cell activation result in altered T cell differentiation and downregulation of immune responses. CTLA4 may regulate signal transduction in a rare subset of CD4+CD25 + T cells which leads to differentiation into regulatory cells. CD40L/CD40 interaction provides a bi-directional signal for T and B cell activation. A possible mechanisms of tolerance induction by CD40L/CD40 blockade involve reduction in expression of B7 molecules, effects on bcl-xL gene and APC function modification. The role of the new discovered pathways: ICOS/B7RP-I and PD-I/PD-LI in regulation of T cell response in transplantation is becoming apparent. PMID- 15759542 TI - New possibilities of therapeutic interventions in transplantation. AB - Immune tolerance of the graft would allow for long-term graft survival without immunosuppressive drugs. Experimental studies showed that both CD28/B7 and CD40L/CD40 costimulations are critical for allograft rejection and their blockade during transplantation induced a decrease of proliferation of alloreactive T cells and an increase of their death by apoptosis. Blocking B7 costimulation increased rodent allograft survival of kidney, liver and pancreatic islets but this is insufficient weapon in the induction of graft tolerance. The combination of the two treatments has synergistic effects. Additional CD40/CD40L blockade represents a adjunct strategy to prevent graft rejection and it has been reported to induce donor specific peripheral tolerance. This therapy prevented acute cardiac allograft rejection and it markedly prolonged allograft survival of kidney and islets in nonhuman primates. A loss of donor-specific alloreactivity has been demonstrated in these experimental models. Calcineurin inhibitors may antagonize the therapeutic effects of costimulatory blockade which may suggest that T cell receptor signaling may be required for tolerance induction. PMID- 15759543 TI - Lung transplantation for end-stage primary pulmonary hypertension. AB - Developments in transplant techniques, peri-operative management and medical therapy have changed the place of transplantation in the treatment armamentarium available for PPH. The aim of this article is therefore, to describe the current knowledge of transplantation for PPH, and to define its place in view of the recent advances in medical therapy. PMID- 15759544 TI - Function of parathyroid glands in kidney transplant patients--diagnostic value of CAP and CIP. AB - OBJECTIVES: As shown in the last four years, Nichols assay for the estimation of "intact" parathyroid hormone (i-PTH) apparently overestimates parathyroid gland function by recognizing both the whole PTH- 1-84 molecule (identified as a cyclase activating PTH - CAP) and N-truncated fragments of PTH-7-84 (identified as a cyclase inactive PTH - CIP). As PTH-1-84 and PTH-7-84 are presumed to show antagonistic effects on calcaemia and bone turnover, we aimed to assess the relationship between PTH-1-84 and PTH-7-84 plasma levels respectively and bone turnover markers in kidney transplant patients. PATIENTS: 52 patients and 17 healthy subjects were examined at least 4 years after renal transplantation. In all subjects the following parameters were assessed: bone mineral density (BMD) of the total body, L2-L4 vertebrae and femoral neck (DEXA), serum total PTH (i PTH) and PTH-1-84 level, as well as the difference between total PTH and PTH-1-84 (reported as PTH-7-84), activity of alkaline phosphatase (AP), serum concentration of collagen type I cross-linked C-telopeptide, osteocalcin (OC), creatinine (creat), 25-OH-D, total calcium (Ca(total)) and phosphorus (P) concentration. RESULTS: Tx patients were characterized by significantly elevated plasma values of all examined parameters except activity of AP and plasma level of Ca, P and 25-OH-D. Both in HS and Tx patients a significant positive correlation was found between plasma concentration of PTH-1-84 and PTH-7-84. In addition in Tx patients both PTH-1-84 and PTH-7-84 showed a significant positive correlation with plasma creatinine, OC, AP and Ctx and a negative one with BMD T score, while in HS PTH-1-84 and PTH-7-84 were positively correlated with OC and AP and negatively with Ca and BMD (borderline significance). CONCLUSION: Presence of highly significant correlations between PTH-1-84 or PTH-7-84 and markers of both osteogenesis and osteolysis respectively is not consistent with a diagnostic superiority of PTH- 1-84 and PTH-7-84 over total PTH estimation in patients 4 or more years after renal transplantation. PMID- 15759545 TI - Plasma neuropeptide Y concentration in kidney transplant patients during the early posttransplant period. AB - OBJECTIVES: Haemodialysed patients with chronic renal failure are characterized by elevated plasma neuropeptide Y (NPY) concentration. Successful kidney transplantation is followed by a regression of the uraemic state and normalization of hormonal and metabolic abnormalities. The aim of the present study was to assess the influence of successful kidney transplantation on plasma NPY concentration in patients with chronic renal failure. METHODS: A total of 38 patients with chronic renal failure was examined after successful kidney transplantation. The control group consisted of 33 healthy subjects. In kidney transplant recipients plasma NPY concentration was assessed immediately before kidney transplantation and additionally two times after successful kidney transplantation: 2-4 days after surgical procedure and just before discharge of the patient from the hospital, when the graft excretory function was satisfactory. At the days specified above blood samples for NPY estimation were withdrawn at 8.00 am, 4.00 pm and 12.00 pm. RESULTS: Plasma NPY concentration in patients before kidney transplantation was significantly (p<0.001) higher than in healthy subjects. Plasma NPY concentration assessed at 8.00 am, 4.00 pm and at 12.00 pm both 2-4 days after kidney transplantation and one day before discharge of the patient from hospital was significantly higher than in healthy subjects (p<0.001). Plasma NPY concentration in kidney transplant patients just before discharge of the patient from hospital did not differ from 2-4 days after surgical procedure. Both in graft recipients and in healthy individuals no significant diurnal changes in NPY plasma concentration were noticed. No significant correlation was found between plasma NPY concentration and blood pressure in kidney transplant patients and in healthy subjects. CONCLUSION: As successful kidney transplantation does not normalize plasma NPY concentration in the early posttransplant period, implies that the transplanted kidney with good excretory functions and immunosupressive therapy are not essential factors involved in the maintence of elevated NPY plasma level in the early posttransplant period. PMID- 15759546 TI - Modality of brain death and serum troponin T levels in donors and their heart transplant recipients. AB - Cardiac troponin T levels and inotropic requirements were assessed in the sera of 39 heart donors immediately before heart harvesting and, in their recipients, during the first two days after transplantation. In the donors who died of cranial trauma (n=21), serum troponin T levels as well as inotropic support were lower (P<0.05 and P<0.025, respectively) than in the donors (n= 18) in whom death was caused by spontaneous intracranial haemorrhage. In the recipients, no differences in troponin T or inotropic support levels were found in relation to the modality of their donors' brain death. PMID- 15759547 TI - Antibodies to HLA class II antigens as a risk factor for acute rejection of the allogeneic kindey. AB - We have investigated the association between the presence of antibodies to HLA class II antigens and the development of acute and chronic rejection after kidney transplantation. Sera from seventy-one patients before, shortly (2 weeks), and in the period between 8 and 22 months after transplantation were analyzed by the standard complement-dependent cytotoxicity (CDC) test, ELISA-LATM, and LAT tests. Absence of antibodies to HLA class II antigens before and shortly after transplantation was associated with a lower incidence of rejection episodes in the first post-transplant year. Donor-specific class II antibodies could not be detected by the ELISA-LAT test and there was no statistically significant difference in serum creatinine levels between the antibody-positive and antibody negative patient groups two years after transplantation. Our study suggests that anti-HLA class II antibodies represent a risk factor for the development of acute immunological complications during the first year after transplantation. PMID- 15759548 TI - Serotonin levels in the small bowel mucosa as a marker of ischemic injury during small bowel preservation. AB - Of the serotonin occurring in the small bowel mucosa, 95% is present in enterochromaffine cells. The cold ischemia during small bowel transplantation results in mucosal injury and releasing of serotonin into the lumen. Because of it, the mucosal concentration of serotonin is decreasing. The aim of our study was to establish the correlation between changes in serotonin levels in small bowel mucosa during grafts preservation and cold ischemic time. Wistar rats (n= 35) weighing 322+/-18g, divided into five main groups (n= 7/group) according to the time of small bowel grafts preservation (0, 1, 6, 9, and 12 hours), were used as experimental animals. The grafts were preserved in 4 degrees C histidine tryptophane-ketoglutarate (HTK) solution. Tissue samples for mucosal serotonin concentration measurement and for light microscopic evaluation were taken after predefined cold ischemic times. Quantitative histological assessment was made using the Park's small bowel wall injury grading scheme. The t-test for dependent samples was used for statistical analysis. The mean serotonin mucosal concentrations after 0, 1, 6, 9, and 12 hours of cold ischemic injury were 433.09+/-160.33, 402.6+/-120.53, 412.5+/-47.57 ng/mL, 190.8+/-45.88 and 145.2+/ 16.78 ng/mL Statistically significant differences (p<0.05) were between 6, 9, and 12 hours of cold ischemia. Morphological changes of small bowel mucosa graded by Park's scheme after the same ischemic intervals were 0, 0.5+/-0.47, 0.97+/-0.41, 1.74+/-0.69, and 1.84+/-0.64. Statistically significant differences (p<0.05) were demonstrated between all preservation times except between 9 and 12 hours of cold ischemia. Morphological changes in small bowel mucosa correlated with cold ischemic time, as well as with serotonin mucosal concentration. These data indicate the possibility of use a serotonin concentration in small bowel mucosa as a parameter of small bowel grafts ischemic injury. PMID- 15759549 TI - Arginase activity concentration marking in monitoring of hepatocytes function after orthotopic liver transplantation--preliminary report. AB - The constant growth of a liver transplantations performed in Poland is the primary reason for a search for new, better and more exact methods of graft function assessment. They should give an opportunity of early detection of either perfusion disorders or organ's function deterioration. Thus, the goal of the study was assessment of the usefulness of arginase concentration marking in liver function monitoring after orthotopic transplantation. The group of 5 recipients of orthotopic liver transplants were taken into account. They were performed in 2003, in the Medical University of Warsaw Department of General, Transplant and Liver Surgery. Patients were qualified for the operation according to standards in the center. The indication for the operation was end-stage liver insufficiency. The average age in the study group was 45,6 years. The patient's serum was taken two hours before the operation. Subsequently, the concentration of arginase activity was marked in serum and bile, first during day 0, 18 hours after the operation, and later at regular intervals during following days until the patient left the clinic, on average from 12 to 14 days. The arithmetical mean values of serum and bile arginase activity values were compared with the average value of a particular parameter, marked during the days following the liver transplant. Serum and bile arginase level fluctuations were assessed in relation to AST, ALT, bilirubin, LDH, alkaline phosphatase (ALKP) and gamma gluthamylo transpeptydase GGTP PMID- 15759550 TI - Clinical significance of arginase after liver transplantation. AB - Liver graft function after transplantation is dependent on ischemia-reperfusion injury, toxicity of drugs (immunosuppression, antibiotics and other) and transplant rejection. Although routinely monitored with enzymatic tests (AST, ALT, GGT, ALP), bilirubin and coagulation parameters, differentiation between these pathologies is hardly possible without liver biopsy. Arginase (3.5.3.1) mostly exists in the liver and in trace amounts in extra-hepatic tissue. Thus, we hypothesized that activity of arginase could be a more specific test of liver function. Sera of 32 liver transplant recipients were tested for AST, ALT, ATIII, bilirubin and arginase. Samples were obtained daily in first 2 weeks after LTx and weekly afterwards. Correlation of arginase activity with other liver function markers was calculated. Serum arginase peaked at day 1 post LTx (mean 64,6+/-91 IU/L), and decreased more rapidly than other tests if good liver function was observed. The values showed strong and significant correlation with AST and ALT activities (Pearsons R 0,65 and 0,47 respectively). We conclude that activity of arginase in the serum is an exact test of liver function. PMID- 15759551 TI - Pregnancies in renal transplant recipients--with a focus on the maternal issues. AB - One hundred and thirteen pregnancies in 73 women who received a renal transplant were studied. The duration between the transplant operation and the conception averaged 19.9 months with a range of I month to 72 months. The mean age was 28 years. The majority (50 recipients had one pregnancy each, but two women had 5 children each and one had seven children). Only 12 (11.5%) out of the 113 pregnancies ended up with spontaneous abortions. The maternal medical problems encountered were reversible rejection in 11%, hypertension in 43%, UTI in 17% and gestational diabetes in 21%. It was noted that there was high incidence of preterm delivery in 64% of the pregnancies Caesarian section was required in 72% % of the cases. In conclusion we observed a high prevalence of successful pregnancies with no adverse effect on the allograft function and excellent fetal outcome (which is described in a separate paper focusing on the fetal outcome). PMID- 15759552 TI - Pregnancies in renal transplant recipients--with a focus on babies. AB - We studied 113 pregnancies in 72 renal transplant recipients. The mean duration between the transplant operation and the first pregnancies was 19.9 months (1 72). This paper focuses on the findings in babies and their outcome. The male babies were 34% of the total. 28% of the babies were delivered vaginally and 72% by Caesarian section. It was noted that there was high incidence of preterm delivery in 64% of the pregnancies. The birth weights were also less than that observed in the general population. 84% was under 50th percentile for weight with 19% being under the 10th percentile. (Saudi growth chart, KACST) The mean hospital stay was 18 days (2-44). The Apgar score was less than 7 in only 10% of the cases and 10 in 50.9%. The incidence of congenital defects was low and no different than the general population (we encountered only 4 cases with minor defects). Despite exposure to cyclosporine throughout the pregnancy and having reduced Nephron mass by virtue of their low birth weight we could not find any glomerular or tubular defects, hypertension or prorienuria in 41 children with a mean age of 52 months. We advised against breast feeding in view of the presence of cyclosporine in the mothers' milk. PMID- 15759553 TI - Application of sulforaphane--does it lead to improvement of islet graft survival after warm and/or cold ischemia. AB - OBJECTIVES: Brain stem death results in ischemic damage of organs. To prevent many agents are being tested this damage. The aim of our study was to determine effects of sulforaphane (SF) on recovery, viability, lipid peroxidation, metabolic and endocrine function of islets isolated from rat pancreata and treated with warm and/or cold ischemia and then transplanted syngeneically. METHODS: Rat pancreata were recovered from non-heart beating rats after intraducatal injection of collagenase solution after 15 or 30 minutes of warm ischemia time (WT). Cold ischemia (CT) was obtained by storage of distended and harvested glands in tubes with UW solution in 4 degrees C for 120 minutes. Sulforaphane was administered 24 hours before isolation islets in concentration 24mg/kg b.w. Diabetes was achieved by intravenouse injection of streptozotocin (STZ 65mg/kg b.w.). Islets were transplanted into the liver through the portal vein. Experimental protocol included four groups: Group I: fresh pancreata not treated with SF, WT=0, CT=0; Group II: 15 or 30 min. of WT, with or without SF; Group III: 15 or 30 min. of WT and 120 min. of CT, with or without SF; Group IV: 120 min. of CT, with or without SF RESULTS: Stimulation index in all groups with sulforaphane was larger from 1.0 (normal response of islets on high concentration glucose in medium). Metabolic activity (MTT) in group II (WT15, SF), gr. IV (CT, SF) and in gr. III (WT15, CT, SF; p>0.05) was lower compared to control group (I). The concentration of MDA in groups with SF increased as compared to controls. The highest recovery and cell viability was observed in group IV (CT, SF) and in gr. III (WT15, CT, SF; p<0.05). In groups exposed to 30 min. of warm ischemia and/or 120 min. of cold preservation was observed higher % of dead islets cells. In vivo study shows that islets graft isolated from rat pancreata treated with sulforaphane reverse diabetes. CONCLUSIONS: Based on results we can conclude that SF in concentration 24mg/kg b.w. reveals protective effect on preserved pancreas and may have a potential clinical implication to improve hemodynamically unstable pancreas donor condition. PMID- 15759554 TI - The use of fascia lata and bone allograft for uncontained defects in revision knee arthroplasty. AB - One of the possibilities in the reconstruction of severe bone loss in revision total knee arthroplasty is impaction bone grafting with support of a mesh. We report the use of a fascia lata allograft as a biological mesh for keeping involved impacted bone graft in one case of uncontained tibial bone defects. PMID- 15759555 TI - Reach: an unprecedented European initiative for regulating industrial chemicals. AB - In February 2001, the European Commission (EC) released a White Paper, REACH 2001, detailing unique and unprecedented legislative proposals for the regulation of industrial chemicals, based on the Precautionary Principle. The object of these proposals was to reverse the escalating incidence of avoidable cancers, a wide range of other industrial diseases, and environmental contamination. However, REACH was aggressively opposed by the European and U.S. chemical industries, and even more so by the U.S. administration. The EC responded by making major concessions in its October 2003 REACH-based legislative proposals. This report critically analyzes REACH, and its 2003 revised proposals, and recommends that REACH be strengthened, not weakened. Furthermore, the report urges that regulatory policies of the United States and other industrialized nations be drastically reformed to comply with those of REACH. PMID- 15759556 TI - Concentration of U.S. hospitals, 1991--1999, and its implications for a national health program. AB - Concentration of hospitals into systems increased from 1991 to 1999. The share of patient care in systems as opposed to independent hospitals rose from 39 to 57 percent. More concentration occurred among secular not-for-profit hospitals than among for-profit hospitals. Systems of hospitals raise the concentration ratio in local markets dramatically. The change in economic structure and market power challenges traditional proposals and strategies aiming to achieve a national health program. PMID- 15759557 TI - Myths and memes about single-payer health insurance in the United States: a rebuttal to conservative claims. AB - Recent years have seen the rapid growth of private think tanks within the neoconservative movement that conduct "policy research" biased to their own agenda. This article provides an evidence-based rebuttal to a 2002 report by one such think tank, the Dallas-based National Center for Policy Analysis (NCPA), which was intended to discredit 20 alleged myths about single-payer national health insurance as a policy option for the United States. Eleven "myths" are rebutted under eight categories: access, cost containment, quality, efficiency, single-payer as solution, control of drug prices, ability to compete abroad (the "business case"), and public support for a single-payer system. Six memes (self replicating ideas that are promulgated without regard to their merits) are identified in the NCPA report. Myths and memes should have no place in the national debate now underway over the future of a failing health care system, and need to be recognized as such and countered by experience and unbiased evidence. PMID- 15759558 TI - Public support for social financing of health care in Switzerland. AB - The purpose of this study was to identify factors associated with the public's preference for financing health care according to people's ability to pay. The authors compared voters' support in 26 Swiss cantons for a legislative proposal to replace regionally rated health insurance premiums (current system) with premiums proportional to income and wealth, and co-financed through the value added tax. The vote took place in May 2003, and the initiative was rejected, with only 27 percent of support nationwide. However, support varied more than threefold, from 13 to 44 percent, among cantons. In multivariate analysis, support was most strongly correlated with the approval rate of the 1994 law on health insurance, which strengthened solidarity between the sick and the healthy. More modest associations were seen between support for the initiative and the health insurance premium of 2003, and proportions of elderly and urban residents in the population. Hence support for more social financing of health care was best explained by past preference for a social health insurance system in the local community. PMID- 15759559 TI - Supplemental analyses of recent trends in infant mortality. AB - U.S. preliminary data for 2002 show a significant increase in the infant mortality rate to 7.0 infant deaths per 1,000 live births, the first rise in the infant mortality rate since 1958. The increase in infant mortality was concentrated in the neonatal period, particularly in deaths occurring within seven days of birth. Partially edited fetal death data suggest that the increase in neonatal mortality was accompanied by a decline in the late fetal mortality rate, and thus it appears that the 2002 perinatal mortality rate will remain level. Potential explanatory factors for the changes in the infant mortality rate are examined, including causes of infant death, percentage of births that are preterm, and low birthweight. Data from the 2002 linked birth and infant death file will allow an assessment of the contribution of maternal and infant factors such as multiple births and management of labor and delivery. PMID- 15759560 TI - Colombia: in vivo test of health sector privatization in the developing world. AB - The reform of the Colombian health sector in 1993 was founded on the internationally advocated paradigm of privatization of health care delivery. Taking into account the lack of empirical evidence for the applicability of this concept to developing countries and the documented experience of failures in other countries, Colombia tried to overcome these problems by a theoretically sound, although complicated, model. Some ten years after the implementation of "Law 100," a review of the literature shows that the proposed goals of universal coverage and equitable access to high-quality care have not been reached. Despite an explosion in costs and a considerable increase in public and private health expenditure, more than 40 percent of the population is still not covered by health insurance, and access to health care proves uncreasingly difficult. Furthermore, key health indicators and disease control programs have deteriorated. These findings confirm the results in other middle- and low-income countries. The authors suggest the explanation lies in the inefficiency of contracting-out, the weak economic, technical, and political capacity of the Colombian government for regulation and control, and the absence of real participation of the poor in decision-making on (health) policies. PMID- 15759561 TI - Vacation-still an issue of workers' protection? An empirical study of vacation and recuperation. AB - The aim of this study was to investigate how commonly employees take vacation or compensatory leave instead of sick leave, and how commonly they do not feel rested/recuperated following a fairly lengthy vacation. In the study group (n = 2,536), consisting of a representative sample based on Sweden's regular labor market surveys, 14 percent of subjects had taken vacation or compensatory leave over the previous 12 months instead of taking sick leave. As many as 15 percent of subjects in the entire sample reported not being rested/recuperated when returning to work following several weeks' leave. A stepwise logistic regression prediction model was constructed to compute the probabilities of not being recuperated. Personal financial situation is a contributory factor, but there is also a strong correlation with the way work is organized. PMID- 15759562 TI - The Sachs report: investing in health for economic development--or increasing the size of the crumbs from the rich man's table? Part II. AB - The Commission on Macroeconomics and Health report (Sachs report of 2001) has been heralded as inspiring and groundbreaking and is being adopted as the blueprint for global health policymaking. This article argues that the report is deeply conservative and unoriginal. It encourages medico-technical solutions to public health problems; it ignores macroeconomic determinants and other root causes of both poor health and poverty; it reverses public health logic and history; it is based on a set of flawed assumptions; it reflects one particular economic perspective to the exclusion of all others; and it recommends greater amounts of charity while preserving the status quo of a deeply unjust and irrational international economic order. Wishful thinking and ideology are presented as established facts to legitimize globalization, and health is conceived primarily as an input to productivity rather than as a human right. The benefits that would result from simple, macroeconomic measures directed toward social justice and the meeting of basic needs are incomparably greater than those that would result from following CMH recommendations in terms of sustainable improvements in both health and economic well-being. The ultimate source of poor health status and miserable living conditions is the extreme concentration of power, nationally and internationally, in the hands of the few. PMID- 15759563 TI - "No one left abandoned": Cuba's national health system since the 1959 revolution. AB - In spite of the economic hardships during the 1990s, Cuba has achieved health indicators that are among the best in the world. This article describes the development of the Cuban health system over more than four decades and analyzes its dynamics. Four stages can be identified. The system's foundations were laid during the first post-revolutionary decade (1959--1970) and consolidated during the succeeding decade (1970--1979). In the third stage, from 1980 onward, the system reached its full expansion with the development of family medicine. Following the crisis of the 1990s, a fourth stage began with reforms and adjustments to the new situation after the collapse of the Soviet Union. Today, health care continues to be of high quality and free for all Cubans. It remains exclusively in the hands of the public sector, and privatization is not an option. This is exactly the opposite of what is happening in other parts of the world where public services are underfunded and people are made to believe that privatization is the only way to ensure high-quality care. PMID- 15759564 TI - On modeling center of foot pressure distortion through a medium. AB - The center of foot pressure (COP) is a commonly used output measure of the postural control system as it is indicative of the systems stability. A dense piece of foam, i.e., a sponge, can be used to emulate random environmental conditions that distort the ground reaction forces received and interpreted by the cutaneous sensors in the feet; thus introducing uncertainty into the control system. In this paper, the density and size of the sponge was selected such that a subject's weight did not cause full compression. In general, the COP is measured from the bottom of the sponge. As the sponge is used to distort ground reaction forces, it is reasonable then to assume that the COP signal would also be distorted. The use of other sensory information to identify state of balance, and compute necessary balance adjustments, is therefore required. In addition to a sponge, many different types of specialized footwear and inserts are used for people with peripheral neuropathy, such as diabetics. However, it is difficult to design diabetic footwear without a better understanding of the mechanical and physiological effects that different surfaces typical of outdoor terrains, such as a sponge, which cannot be predicted without the sense of the foot, have on balance. Therefore, the goal of this study was to investigate the change of the COP signal from the top and bottom of the sponge. Portable force sensing mats from Vista Medical were used to obtain the COP from the top and bottom of the sponge. The COP measured on the bottom of the sponge is not the same as the COP measured on the top, particularly in the medial-lateral direction. Several linear and nonlinear models were used to identify the unknown plant; i.e., the sponge. Overall, the nonlinear neural network method had superior performance when compared with the linear models. Thus, the results indicate that the signals from the top and bottom of the sponge are in fact different, and furthermore, they are nonlinearly related. A nonlinear mathematical model is proposed which describes COP distortion through a medium such as a sponge. Although the values for the model parameters determined were for a particular sponge, this study suggests that a neural network plant identification model may be applied to any medium other than the sponge; the information can then be used to determine how the balance control model is affected given the sensory information received. PMID- 15759565 TI - Nonlinear control of a dynamic model of HIV-1. AB - Highly active antiretroviral therapy (HAART) reduces the viral burden in human immunodeficiency virus type 1 (HIV-1) infected patients. The paper addresses the problem of controlling the predator-prey like model of the interaction among CD4+ T-cell, CD8+ T-cell, and HIV-1 by an external drug agency. By exploring the dynamic properties of the system, the original system is first regrouped into two subsystems, then a nonlinear global controller is presented by designing two controllers over two complementary zones: a local controller on a finite region and a global controller over its complement. The local controller is designed to guarantee nonnegativty, and avoids the problem of control singularity within the neighborhood of the origin omega. The complementary controller is designed via backstepping for both subsystems over the complementary region. The closed-loop system is globally stable at nominal values through the introduction of a novel bridging virtual control, and the resulting controller is singularity free and guarantees nonnegativity. In this paper, simulations are conducted in discrete time with sampling time Ts to show the effectiveness of the proposed method. PMID- 15759566 TI - Localized electrical nerve blocking. AB - Localized electrical nerve blocking was investigated in computer simulation and in vivo trials for sinusoidal frequencies between 5 and 20 kHz. Computer simulations indicated that a localized transmission block of the axons could occur in each of the axon models. An approximation of nerve stimulation was derived from individual axon simulations conducted over axon diameters of 5-15 microm and electrode to axon distances of 0.25 to 2.0 mm. Examination of the membrane voltage and ionic gate potentials indicated that the block could be attributed to an elevated membrane voltage. The elevated membrane voltage could prevent conduction of action potentials through the region of the sinusoidal currents. At lower amplitudes, the sinusoidal current could stimulate the axon and generate a continuous series of action potentials. In vivo trials demonstrated that the sinusoidal frequencies of greater than 10 kHz would cause a localized block in rats. Sinusoidal frequencies below 5 kHz would lead to a reduction in muscle force that appeared to be caused by depletion of transmitter at the neuromuscular junction. As indicated by the computer models of rat nerves, the endplate depletion block occurred at a lower frequency (below 5 kHz) than the block (above 10 kHz). A partial block of the axon was demonstrated, suggesting that sinusoidal currents could be used to provide selective stimulation if they are combined with distal electrical stimulation. PMID- 15759567 TI - Toward the neurocomputer: image processing and pattern recognition with neuronal cultures. AB - Information processing in the nervous system is based on parallel computation, adaptation and learning. These features cannot be easily implemented on conventional silicon devices. In order to obtain a better insight of how neurons process information, we have explored the possibility of using biological neurons as parallel and adaptable computing elements for image processing and pattern recognition. Commercially available multielectrode arrays (MEAs) were used to record and stimulate the electrical activity from neuronal cultures. By mapping digital images, i.e., arrays of pixels, into the stimulation of neuronal cultures, a low and bandpass filtering of images could be quickly and easily obtained. Responses to specific spatial patterns of stimulation were potentiated by an appropriate training (tetanization). Learning allowed pattern recognition and extraction of spatial features in processed images. Therefore, neurocomputers, (i.e., hybrid devices containing man-made elements and natural neurons) seem feasible and may become a new generation of computing devices, to be developed by a synergy of Neuroscience and Material Science. PMID- 15759568 TI - Multiregion bicentric-spheres models of the head for the simulation of bioelectric phenomena. AB - Equations are derived for the electric potentials [electroencephalogram (EEG)] produced by dipolar sources in a multiregion bicentric-spheres volume-conductor head model. Being the equations valid for an arbitrary number of regions, our proposal is a generalization of many spherical models presented so far in literature, each of those regarded as a particular case of our multiregion model. Moreover, our approach allows considering new features of the head volume conductor to better approximate electrical properties of the real head. PMID- 15759569 TI - Fetal electrocardiogram extraction by sequential source separation in the wavelet domain. AB - This paper addresses the problem of fetal electrocardiogram extraction using blind source separation (BSS) in the wavelet domain. A new approach is proposed, which is particularly advantageous when the mixing environment is noisy and time varying, and that is shown, analytically and in simulation, to improve the convergence rate of the natural gradient algorithm. The distribution of the wavelet coefficients of the source signals is then modeled by a generalized Gaussian probability density, whereby in the time-scale domain the problem of selecting appropriate nonlinearities when separating mixtures of both sub- and super-Gaussian signals is mitigated, as shown by experimental results. PMID- 15759570 TI - Statistical maps for EEG dipolar source localization. AB - We present a method that estimates three-dimensional statistical maps for electroencephalogram (EEG) source localization. The maps assess the likelihood that a point in the brain contains a dipolar source, under the hypothesis of one, two or three activated sources. This is achieved by examining all combinations of one to three dipoles on a coarse grid and attributing to each combination a score based on an F statistic. The probability density function of the statistic under the null hypothesis is estimated nonparametrically, using bootstrap resampling. A theoretical F distribution is then fitted to the empirical distribution in order to allow correction for multiple comparisons. The maps allow for the systematic exploration of the solution space for dipolar sources. They permit to test whether the data support a given solution. They do not rely on the assumption of uncorrelated source time courses. They can be compared to other statistical parametric maps such as those used in functional magnetic resonance imaging (fMRI). Results are presented for both simulated and real data. The maps were compared with LORETA and MUSIC results. For the real data consisting of an average of epileptic spikes, we observed good agreement between the EEG statistical maps, intracranial EEG recordings, and fMRI activations. PMID- 15759571 TI - Model selection in spatio-temporal electromagnetic source analysis. AB - Several methods [model selection procedures (MSPs)] to determine the number of sources in electroencephalogram (EEG) and magnetoencphalogram (MEG) data have previously been investigated in an instantaneous analysis. In this paper, these MSPs are extended to a spatio-temporal analysis if possible. It is seen that the residual variance (RV) tends to overestimate the number of sources. The Akaike information criterion (AIC) and the Wald test on amplitudes (WA) and the Wald test on locations (WL) have the highest probabilities of selecting the correct number of sources. The WA has the advantage that it offers the opportunity to test which source is active at which time sample. PMID- 15759572 TI - Discrimination of pathological voices using a time-frequency approach. AB - Acoustical measures of vocal function are routinely used in the assessments of disordered voice, and for monitoring the patient's progress over the course of voice therapy. Typically, acoustic measures are extracted from sustained vowel stimuli where short-term and long-term perturbations in fundamental frequency and intensity, and the level of "glottal noise" are used to characterize the vocal function. However, acoustic measures extracted from continuous speech samples may well be required for accurate prediction of abnormal voice quality that is relevant to the client's "real world" experience. In contrast with sustained vowel research, there is relatively sparse literature on the effectiveness of acoustic measures extracted from continuous speech samples. This is partially due to the challenge of segmenting the speech signal into voiced, unvoiced, and silence periods before features can be extracted for vocal function characterization. In this paper we propose a joint time-frequency approach for classifying pathological voices using continuous speech signals that obviates the need for such segmentation. The speech signals were decomposed using an adaptive time-frequency transform algorithm, and several features such as the octave max, octave mean, energy ratio, length ratio, and frequency ratio were extracted from the decomposition parameters and analyzed using statistical pattern classification techniques. Experiments with a database consisting of continuous speech samples from 51 normal and 161 pathological talkers yielded a classification accuracy of 93.4%. PMID- 15759573 TI - A least-squares parameter estimation algorithm for switched hammerstein systems with applications to the VOR. AB - A "Multimode" or "switched" system is one that switches between various modes of operation. When a switch occurs from one mode to another, a discontinuity may result followed by a smooth evolution under the new regime. Characterizing the switching behavior of these systems is not well understood and, therefore, identification of multimode systems typically requires a preprocessing step to classify the observed data according to a mode of operation. A further consequence of the switched nature of these systems is that data available for parameter estimation of any subsystem may be inadequate. As such, identification and parameter estimation of multimode systems remains an unresolved problem. In this paper, we 1) show that the NARMAX model structure can be used to describe the impulsive-smooth behavior of switched systems, 2) propose a modified extended least squares (MELS) algorithm to estimate the coefficients of such models, and 3) demonstrate its applicability to simulated and real data from the Vestibulo Ocular Reflex (VOR). The approach will also allow the identification of other nonlinear bio-systems, suspected of containing "hard" nonlinearities. PMID- 15759574 TI - A model order selection criterion with applications to cardio-respiratory-renal systems. AB - We introduce a model order selection criterion called signal prediction error (SPE) for the identification of a linear regression model, which can be an adequate representation of a resting physiologic system. SPE is an estimate of the prediction error variance due only to model estimation error and not unobserved noise, which distinguishes it from the widely used final prediction error (FPE). We then present a theoretical analysis of SPE, which predicts that its ability to select correctly the model order is more dependent on the signal to-noise ratio (SNR) and less dependent on the number of data samples available for analysis. We next propose a heuristic procedure based on SPE (called SPE(D)) to improve its robustness to SNR levels. We then demonstrate, through simulated physiologic data at high SNR levels, that SPE will be equivalent to consistent model order selection criteria for long data records but will become superior to FPE and other model order selection criteria as the size of the data record decreases. The simulated data results also show that SPE(D) is indeed a significant improvement over SPE in terms of robustness to SNR. Finally, we demonstrate the applicability of SPE and SPE(D) to actual cardio-respiratory renal data. PMID- 15759576 TI - Composition of approximated body-surface-potential-maps by utilizing a common 12 lead-ECG device. AB - A procedure is introduced that allows approximations of body surface potential maps (BSPM) to be obtained by utilizing commonly available digital 12-lead electrocardiogram (ECG) systems. These Pseudo-BSPMs contain most of the averaged spatio-temporal information for a single characteristic beat. The underlying signal processing is described in detail. The algorithms including an online method verification may easily be added to the software of commercial 12-lead ECG devices. PMID- 15759575 TI - Robust time delay estimation of bioelectric signals using least absolute deviation neural network. AB - The time delay estimation (TDE) is an important issue in modern signal processing and it has found extensive applications in the spatial propagation feature extraction of biomedical signals as well. Due to the extreme complexity and variability of the underlying systems, biomedical signals are usually nonstationary, unstable and even chaotic. Furthermore, due to the limitations of the measurement environments, biomedical signals are often noise-contaminated. Therefore, the TDE of biomedical signals is a challenging issue. A new TDE algorithm based on the least absolute deviation neural network (LADNN) and its application experiments are presented in this paper. The LADNN is the neural implementation of the least absolute deviation (LAD) optimization model, also called unconstrained minimum L1-norm model, with a theoretically proven global convergence. In the proposed LADNN-based TDE algorithm, a given signal is modeled using the moving average (MA) model. The MA parameters are estimated by using the LADNN and the time delay corresponds to the time index at which the MA coefficients have a peak. Due to the excellent features of L1-norm model superior to Lp-norm (p > 1) models in non-Gaussian noise environments or even in chaos, especially for signals that contain sharp transitions (such as biomedical signals with spiky series or motion artifacts) or chaotic dynamic processes, the LADNN based TDE is more robust than the existing TDE algorithms based on wavelet-domain correlation and those based on higher-order spectra (HOS). Unlike these conventional methods, especially the current state-of-the-art HOS-based TDE, the LADNN-based method is free of the assumption that the signal is non-Gaussian and the noises are Gaussian and, thus, it is more applicable in real situations. Simulation experiments under three different noise environments, Gaussian, non Gaussian and chaotic, are conducted to compare the proposed TDE method with the existing HOS-based method. Real application experiment is conducted to extract time delay information between every two adjacent channels of gastric myoelectrical activity (GMA) to assess the spatial propagation characteristics of GMA during different phases of the migrating myoelectrical complex (MMC). PMID- 15759577 TI - Distinguishing between moving and stationary sources using EEG/MEG measurements with an application to epilepsy. AB - Performances of electroencephalography (EEG) and magnetoencephalography (MEG) source estimation methods depend on the validity of the assumed model. In many cases, the model structure is related to physical information. We discuss a number of statistical selection methods to distinguish between two possible models using least-squares estimation and assuming a spherical head model. The first model has a single moving source whereas the second has two stationary sources; these may result in similar EEG/MEG measurements. The need to decide between such models occurs for example in Jacksonian seizures (e.g., epilepsy) or in intralobular activities, where a model with either two stationary dipole sources or a single moving dipole source may be possible. We also show that all of the selection methods discussed choose the correct model with probability one when the number of trials goes to infinity. Finally we present numerical examples and compare the performances of the methods by varying parameters such as the signal-to-noise ratio, source depth, and separation of sources, and also apply the methods to real MEG data for epilepsy. PMID- 15759578 TI - An observer study methodology for evaluating detection of motion abnormalities in gated myocardial perfusion SPECT. AB - To address the task of detecting nonischemic motion abnormalities from animated displays of gated myocardial perfusion single photon emission computed tomography data, we performed an observer study to evaluate the difference in detection performance between gating to 8 and 16 frames. Images were created from the NCAT mathematical phantom with a realistic heart simulating hypokinetic motion in the left lateral wall. Realistic noise-free projection data were simulated for both normal and defective hearts to obtain 16 frames for the cardiac cycle. Poisson noise was then simulated for each frame to create 50 realizations of each heart, All datasets were processed in two ways: reconstructed as a 16-frame set, and collapsed to 8 frames and reconstructed. Ten observers viewed the cardiac images animated with a realistic real-time frame rate. Observers trained on 100 images and tested on 100 images, rating their confidence on the presence of a motion defect on a continuous scale. None of the observers showed a significant difference in performance between the two gating methods. The 95% confidence interval on the difference in areas under the ROC curve (Az8 - Az16) was -0.029 0.085. Our test did not find a significant difference in detection performance between 8-frame gating and 16-frame gating. We conclude that, for the task of detecting abnormal motion, increasing the number of gated frames from 8 to 16 offers no apparent advantage. PMID- 15759579 TI - Assessment of walking features from foot inertial sensing. AB - An ambulatory monitoring system is developed for the estimation of spatio temporal gait parameters. The inertial measurement unit embedded in the system is composed of one biaxial accelerometer and one rate gyroscope, and it reconstructs the sagittal trajectory of a sensed point on the instep of the foot. A gait phase segmentation procedure is devised to determine temporal gait parameters, including stride time and relative stance; the procedure allows to define the time intervals needed for carrying an efficient implementation of the strapdown integration, which allows to estimate stride length, walking speed, and incline. The measurement accuracy of walking speed and inclines assessments is evaluated by experiments carried on adult healthy subjects walking on a motorized treadmill. Root-mean-square errors less than 0.18 km/h (speed) and 1.52% (incline) are obtained for tested speeds and inclines varying in the intervals [3, 6] km/h and [-5, + 15]%, respectively. Based on the results of these experiments, it is concluded that foot inertial sensing is a promising tool for the reliable identification of subsequent gait cycles and the accurate assessment of walking speed and incline. PMID- 15759581 TI - A clinical water-coated antenna applicator for MR-controlled deep-body hyperthermia: a comparison of calculated and measured 3-D temperature data sets. AB - A magnetic resonance (MR)-compatible three-dimensional (3-D) hyperthermia applicator was developed and evaluated in the magnetic resonance (MR) tomograph Siemens MAGNETOM Symphony 1.5 T. Radiating elements of this applicator are 12 so called water coated antenna (WACOA) modules, which are designed as specially shaped and adjustable dipole structures in hermetically closed cassettes that are filled by deionized water. The WACOA modules are arranged in the applicator frame in two transversal antenna subarrays, six antennas per subarray. As a standard load for the applicator an inhomogeneous phantom was fabricated. Details of applicator's realization are presented and a 3-D comparison of calculated and measured temperature data sets is made. A fair agreement is achieved that demonstrates the numerically supported applicator's ability of phase-defined 3-D pattern steering. Further refinement of numerical models and measuring methods is necessary. The applicator's design and the E-field calculations were performed using the finite-difference time-domain (FDTD) method. The calculation and optimization of temperature patterns was obtained using the finite element method (FEM). For MR temperature measurements the proton resonance frequency (PRF) method was used. PMID- 15759582 TI - Implantable cardiac pacemaker electromagnetic compatibility testing in a novel security system simulator. AB - This paper describes a novel simulator to perform electromagnetic compatibility (EMC) tests for active implantable medical devices (AIMDs) with electromagnetic fields emitted by security systems. The security system simulator was developed in response to over 100 incident reports over 17 years related to the interference of AIMD's with security systems and the lack of a standardized test method. The simulator was evaluated regarding field homogeneity, signal distortion, and maximum magnetic field strength levels. Small three-axis probes and a three-axis scanning system were designed to determine the spatial and temporal characteristics of the fields emitted by 12 different types of walk through metal detectors (WTMDs). Tests were performed on four implanted pacemakers with a saline phantom and correlated to a newly developed test method performed "in air" (without the phantom). Comparison of the simulator thresholds with tests performed in real WTMDs showed that the simulator is able to mimic the pacemaker interference. The interference thresholds found in the simulator indicate that pulsed magnetic fields are more likely to cause interference in pacemakers than sinusoidal fields. The security system simulator will help biomedical engineers, manufacturers of medical devices, and manufacturers of security systems to identify incompatible combinations of WTMDs and AIMDs early in the development stage. PMID- 15759580 TI - An inverted-microstrip resonator for human head proton MR imaging at 7 tesla. AB - As an extension of the previously developed microstrip transmission line (MTL) RF coil design, a high-frequency RF volume coil using multiple inverted MTL (iMTL) resonators for human head imaging at high magnetic field strength of 7 tesla (T) is reported. Compared to conventional MTL resonators, iMTL resonators can operate at higher frequency with lower losses and, thus, are suitable for designs of high frequency RF volume coils with large coil size for human MR imaging and spectroscopy at high fields. An approach using capacitive terminations was analyzed and applied to the design of the iMTL volume coil for improving RF field homogeneity and broadening frequency-tuning range. A performance-comparison study was conducted between the prototype iMTL volume coil and a custom-built TEM volume coil at 7 T. The iMTL volume coil presents a comparable SNR and intrinsic B1 homogeneity to the TEM volume coil. Phantom and the human head images acquired using the iMTL volume coil are also presented. The proposed iMTL volume coil provides an efficient and alternative solution to design high-frequency and large size volume coils for human MR applications at very high fields. PMID- 15759583 TI - Stimulation of isolated ventricular myocytes within an open architecture microarray. AB - This paper is concerned with the physiological responses of single heart cells within microfluidic chambers, in response to stimulation by integrated microelectrodes. To enable these investigations, which included the measurement of action potential duration, intracellular Ca2+ and cell shortening, a series of microfluidic chambers (50 microm wide, 180 microm long, 400 microm high, 500 microm pitch) and connecting channels (200 microm wide, 5000 microm long, 50 microm high, 500 microm pitch) were replica-moulded into the silicone elastomer, polydimethylsiloxane (PDMS). The structures were formed against a master of posts and lines, photolithograhically patterned into the high aspect ratio photoresist SU-8. The chambers within the slab of PDMS were aligned against pairs of stimulating gold microelectrodes (50 microm long, 20 microm wide, 0.1-10 microm thick, 180 microm apart) patterned on a microscope coverslip base, thus defining cavities of approximately 4 nL volume. The assembly was filled with physiological saline and single isolated rabbit ventricular myocytes were introduced by micropipetting, thus creating limited volumes of saline above individual myocytes that could be varied between 4 nL and > or = 4 microL. The application of transient current pulses to the cells via the electrodes caused transient contractions with constant amplitude (recorded as changes in sarcomere length), confirming that excitation contraction coupling (EC coupling) remained functional in these limited volumes. Continuous monitoring of the intracellular Ca2+ (using calcium sensitive dyes) showed, that in the absence of bath perfusion, the amplitude of the transients remained constant for approximately 3 min in the 4-nL volume and approximately 20 min for the 4 microL volume. Beyond this time, the cells became unexcitable until the bath was renewed. The action potential duration (APD) was recorded at stimulation frequencies of 1 Hz and 0.5 Hz using potential sensitive dyes and was prolonged at the higher pacing rate. These studies show the prolonged electrical stimulation of isolated adult cardiac myocytes in microchambers with unimpaired EC coupling as verified on optical records of the action potential, Ca2+ transients and cell shortening. The open architecture provided free (pipetting) access for drug dispensation without cross talk between neighboring microwells, and multiplexed optical detection can be realized to study EC coupling on arrays of cells under both control and experimental conditions. PMID- 15759584 TI - Wavelet-based lossy-to-lossless ECG compression in a unified vector quantization framework. AB - In a prior work, a wavelet-based vector quantization (VQ) approach was proposed to perform lossy compression of electrocardiogram (ECG) signals. In this paper, we investigate and fix its coding inefficiency problem in lossless compression and extend it to allow both lossy and lossless compression in a unified coding framework. The well-known 9/7 filters and 5/3 integer filters are used to implement the wavelet transform (WT) for lossy and lossless compression, respectively. The codebook updating mechanism, originally designed for lossy compression, is modified to allow lossless compression as well. In addition, a new and cost-effective coding strategy is proposed to enhance the coding efficiency of set partitioning in hierarchical tree (SPIHT) at the less significant bit representation of a WT coefficient. ECG records from the MIT/BIH Arrhythmia and European ST-T Databases are selected as test data. In terms of the coding efficiency for lossless compression, experimental results show that the proposed codec improves the direct SPIHT approach and the prior work by about 33% and 26%, respectively. PMID- 15759585 TI - Three-dimensional trajectory assessment of an IVUS transducer from single-plane cineangiograms: a phantom study. AB - The recovery of the three-dimensional (3-D) path of the transducer used during an intravascular ultrasound (IVUS) examination is of primary importance to assess the exact 3-D shape of the vessel under study. Traditionally, the reconstruction is done by simply stacking the images during the pullback, or more recently using biplane angiography to recover the vessel curvature. In this paper, we explain, how single-plane angiography can be used with two projection models, to perform this task. Two types of projection geometry are analyzed: weak-perspective and full-perspective. In weak-perspective projection geometry, the catheter path can be reconstructed without prior transducer depth information. With full perspective projection geometry, precise depth location of reference points are needed in order to minimize the error of the recovered transducer angle of incidence. The transducer angulation reconstruction is based on the foreshortening effect as seen from the X-ray images. By comparing the measured to the true transducer length, we are able to get its incidence angle. The transducer trajectory is reconstructed by stitching together the different estimated angulations obtained from each image in a cineangiogram sequence. The method is described and validated on two helical vessel phantoms, giving on average a reconstructed path that is less than 2 mm distant from the true path when using full-perspective projection. PMID- 15759586 TI - Comparative analysis of hematocrit measurements by dielectric and impedance techniques. AB - In a previous paper, a new dielectric technique was used to estimate hematocrit (HTC) in extracorporeal blood circulation systems, independently of plasma conductivity or osmolarity. Although many impedance techniques have been formerly proposed in the literature, none has been evaluated against plasma conductivity and osmolarity. Herein, we estimate HTC based on permittivity changes and also with other four techniques found in the literature. Besides, the error incurred in each is also studied when plasma conductivity and osmolarity changed as much as 1 mS/cm and 50 mOsm/kg, respectively. The dielectric (permittivity) technique has an error close to 5.4%, while the others showed both tendencies, i.e., lower error (2.5%, two of them) and higher error (8.6% and 16.3%, the other two). The dielectric technique, even though did not produce the lowest error, provides a well-described physical model along with simple instrumentation. PMID- 15759587 TI - The relations among maternal depression, maternal criticism, and adolescents' externalizing and internalizing symptoms. AB - This study examined the relations between maternal criticism and externalizing and internalizing symptoms in adolescents who varied in their risk for psychopathology. Both maternal-effects and child-effects models were examined. The sample consisted of 194 adolescents (mean age = 11.8 years) and their mothers; 146 mothers had a history of depressive disorders and 48 did not. When adolescents were in 6th and 8th grade, maternal criticism was measured with the five-minute speech sample and adolescents' symptoms were assessed with the Child Behavior Checklist. Maternal criticism was significantly associated with both adolescents' externalizing and internalizing symptoms, beyond the contribution of the chronicity/severity of mothers' depression history. Maternal criticism did not mediate the relation between maternal depression and adolescent symptoms. In contrast, adolescent externalizing behaviors mediated the relation between chronicity/severity of maternal depression history and maternal criticism in 6th grade. Prospective analyses showed that adolescents' externalizing symptoms in 6th grade significantly predicted maternal criticism in 8th grade, controlling for maternal depression history and prior maternal criticism. Results are discussed in terms of the importance of examining child-effects models in studies of maternal criticism. PMID- 15759588 TI - Early adversity and the prospective prediction of depressive and anxiety disorders in adolescents. AB - The current study was a prospective exploration of the specificity of early childhood adversities as predictors of anxiety and depressive disorders in adolescents. Participants were 816 adolescents (414 males, 402 females) with diagnostic information collected at age 15; information on early adversities had been collected from the mothers during pregnancy, at birth, age 6 months, and age 5 years for a related study. Adolescents with "pure" anxiety disorders were compared with adolescents with "pure" depressive disorders (major depressive disorder, dysthymia), and these groups were compared to never-ill controls. Analyses controlled for gender and maternal depression and anxiety disorders. Results indicated that adolescents with anxiety disorders were more likely than depressed youth to have been exposed to various early stressors, such as maternal prenatal stress, multiple maternal partner changes, and more total adversities, whereas few early childhood variables predicted depressive disorders. Even when current family stressors at age 15 were controlled, early adversity variables again significantly predicted anxiety disorders. Results suggest that anxiety disorders may be more strongly related to early stress exposure, while depressive disorders may be related to more proximal stressors or to early stressors not assessed in the current study. PMID- 15759590 TI - Pathways to adolescent substance use among sexually abused girls. AB - This study examined the link between childhood sexual abuse and adolescent substance use among girls, and evaluated depressive self-concept and behavioral under-control (BUC) as pathways to substance use for sexually abused girls. Participants (n = 150) were drawn from a longitudinal study of the impact of domestic violence on the lives of women and children. Structural equation modeling revealed that girls' childhood sexual abuse was associated prospectively with their later substance use. This relationship persisted when age, co occurring forms of child abuse (physical, exposure to domestic violence), childhood depression and aggression, family income, maternal substance use, and parenting practices were controlled. Behavioral under-control mediated the relationship between childhood sexual abuse and later substance use, but depressive self-concept did not. Implications, limitations, and directions for future research are discussed. PMID- 15759589 TI - Peer contagion of depressogenic attributional styles among adolescents: a longitudinal study. AB - This study examined longitudinal associations between adolescents' and their friend's depressive symptoms and depressogenic attributional style. Participants included 398 adolescents in grades six through eight at the outset of the study. Adolescents completed peer nominations to identify reciprocated and unreciprocated best friendships as well as measures of depressive symptoms and depressogenic attributional style at an initial time point, and again 11 months later. Results revealed that best friends' reported level of depressive symptoms was prospectively associated with adolescents' own depressive symptoms and with adolescents' depressogenic attributional style. Moderator effects suggested that friends' attributional styles were prospectively associated with adolescents' own attributional styles for those involved in reciprocated friendships. Lastly, findings offered preliminary support for adolescents' Time 2 depressive symptoms as a mediator of the association between friends' depressive symptoms and adolescents' attributional style. Findings have important implications for cognitive and interpersonal models of adolescent depression, as well as the study of peer contagion effects. PMID- 15759591 TI - Understanding teasing: lessons from children with autism. AB - Teasing requires the ability to understand intention, nonliteral communication, pretense, and social context. Children with autism experience difficulty with such skills, and consequently, are expected to have difficulty with teasing. To better understand teasing concepts and behaviors, children with autism, their parents, and age and Verbal-IQ-matched comparison children and parents described concepts and experiences of teasing and engaged in a parent-child teasing interaction. The teasing of children with autism was less playful and provocative and focused less on social norms than that of comparison children. Similarly, parents of children with autism teased in less playful ways. Scores on a theory of mind task accounted for several of the observed differences. Discussion focused on the importance of understanding social context and playful behavior during teasing. PMID- 15759592 TI - Which executive functioning deficits are associated with AD/HD, ODD/CD and comorbid AD/HD+ODD/CD? AB - This study investigated (1) whether attention deficit/hyperactivity disorder (AD/HD) is associated with executive functioning (EF) deficits while controlling for oppositional defiant disorder/conduct disorder (ODD/CD), (2) whether ODD/CD is associated with EF deficits while controlling for AD/HD, and (3) whether a combination of AD/HD and ODD/CD is associated with EF deficits (and the possibility that there is no association between EF deficits and AD/HD or ODD/CD in isolation). Subjects were 99 children ages 6-12 years. Three putative domains of EF were investigated using well-validated tests: verbal fluency, working memory, and planning. Independent of ODD/CD, AD/HD was associated with deficits in planning and working memory, but not in verbal fluency. Only teacher rated AD/HD, but not parent rated AD/HD, significantly contributed to the prediction of EF task performance. No EF deficits were associated with ODD/CD. The presence of comorbid AD/HD accounts for the EF deficits in children with comorbid AD/HD+ODD/CD. These results suggest that EF deficits are unique to AD/HD and support the model proposed by R. A. Barkley (1997). PMID- 15759593 TI - Mother-son interactions in families of boys with Attention-Deficit/Hyperactivity Disorder with and without oppositional behavior. AB - Parenting responsiveness and over-reactivity were assessed among 25 mothers of 7 9-year-old sons with Attention-Deficit/Hyperactivity Disorder (ADHD) and oppositional behavior (Oppositional Defiant, OD), 24 mothers of sons with ADHD only, and 38 mothers of nonproblem sons. Responsiveness was observed during mother-son play and clean-up interactions and over-reactivity was assessed using self-reports of parenting in discipline situations. Mothers of sons with ADHD/OD were less responsive and more over-reactive than mothers of nonproblem sons, and mothers of sons with ADHD only did not differ from the other groups. Mothers of sons with ADHD/OD reported more hostility than mothers of nonproblem sons, and controlling for maternal hostility eliminated the significant group differences in responsiveness during clean-up and in over-reactivity. Controlling for the child's ADHD subtype did not alter the pattern of results. The implications for addressing responsiveness and over-reactivity as components of parent-mediated behavioral treatments for ADHD are considered. PMID- 15759594 TI - Trajectories of peer-nominated aggression: risk status, predictors and outcomes. AB - Developmental trajectories of peer-nominated aggression, risk factors at baseline, and outcomes were studied. Peer nominations of aggression were obtained annually from grades 1 to 3. Three developmental trajectories were identified: an early-onset/increasers trajectory with high levels of peer-nominated aggression at elementary school entry and increasing levels throughout follow-up; a moderate persistent trajectory of aggression in which children were characterized by moderate levels of physical aggression at baseline; and a third trajectory with stable low levels of aggression. Children following the early-onset/increasers trajectory showed physical forms of aggression at baseline. Male gender and comorbid attention deficit/hyperactivity problems, oppositional defiant problems and poor prosocial behavior plus negative life events predicted which children would follow the early-onset/increasers trajectory of aggression. The outcomes associated with the early-onset/increaser children suggest high risk for chronically high levels of aggressive behavior. PMID- 15759595 TI - The etiology of behavior problems in 7-year-old twins: substantial genetic influence and negligible shared environmental influence for parent ratings and ratings by same and different teachers. AB - Parent ratings of behavior problems in childhood show substantial genetic influence and modest shared environmental influence. However, few studies have compared these results to teacher ratings and no previous studies have compared same-teacher ratings to different-teacher ratings. 3,714 7-year-old twin pairs in the Twins Early Development Study were rated by parents and teachers on the Strengths and Difficulties Questionnaire. Substantial heritability and negligible shared environmental influence were found for data from all three raters for total behavior problems, hyperactivity, prosocial behavior, peer problems, conduct problems, and emotional symptoms. Sex-limitation models revealed similar results for males and females, although there was some evidence for greater heritability for boys, especially when twins were rated by the same teacher. PMID- 15759596 TI - Timing of conditioned responses utilizing electrical stimulation in the region of the interpositus nucleus as a CS. AB - A large body of evidence indicates that the cerebellum is essential for the acquisition, retention, and expression of the standard delay conditioned eyeblink response and that the basic memory trace appears to be established in the anterior interpositus nucleus (IP). Adaptive timing of the conditioned response (CR) is a prominent feature of classical conditioning-the CR peaks at the time of onset of the unconditioned stimulus (US) over a wide range of CS-US interstimulus intervals (ISI). A key issue is whether this timing is established by the cerebellar circuitry or prior to the cerebellum. In this study timing of conditioned eyeblink responses established via electrical stimulation of the interpositus nucleus as a conditioned stimulus (CS) was analyzed prior to and following modification of the CS-US interval in well-trained rabbits. Consistent with previous results, learning under these conditions is very rapid and robust. The CR peak eyeblink latencies are initially timed to the US onset and adjust accordingly to lengthening or shortening of the CS-US interval, just as with peripheral CSs. The acquisition of conditioned eyeblink responses by direct electrical stimulation of the IP as a CS thus retains temporal flexibility following shifts in the CS-US delay, as found in standard classical eyeblink conditioning procedures. PMID- 15759597 TI - Conditioned ethanol aversion in rats induced by voluntary wheel running, forced swimming, and electric shock: an implication for aversion therapy of alcoholism. AB - This study was planned to demonstrate rats' acquisition of aversion to ethanol solution consumed before voluntary running, forced swimming, or electric shock delivery. Wistar rats under water deprivation were allotted to four groups of eight rats each, and all rats were allowed to drink 5% ethanol solution for 15 min. Immediately after the ethanol drinking, rats of Group Run were put into the individual running wheels for 15 min, those of Group Swim were put into the individual swimming pools for 15 min, those of Group Shock received electric shocks for 15 min (15 0.45-mA shocks of 0.7s with the intershock interval of 1 min) in the individual small chambers, and those of Group Control were directly returned back to the home cages. This procedure was repeated for six days, followed by a two-day choice test of ethanol aversion where a bottle containing 5% ethanol solution and a bottle of tap water were simultaneously presented for 15 min. In the test, Groups Run, Swim, and Shock drank ethanol solution significantly less than tapwater, while Group Control drank both fluids equally. The effects of running, swimming, and shock were equivalent. The successful demonstration of acquired ethanol aversion induced by exercise (running and swimming) or shock in rats suggests an avenue for clinical application of exercise and shock treatments for human alcoholics, though there are many issues to be resolved before the practical use. PMID- 15759598 TI - Consistency in physiological stress responses and electromyographic activity during induced stress exposure in women and men. AB - Physiological responses serve the role as objective indicators of stress as well as a link between psychosocial stress and various health outcomes. The aim of the present exposure session was to compare different physiological stress responses (systolic and diastolic blood pressure, heart rate, urinary epinephrine and norepinephrine, salivary cortisol) as well as trapezius muscle activity, measured by surface electromyography, during mental and physical stress in 11 women and ten men. The results show significantly increased activity in all measures but cortisol and significant associations between sympathetic arousal and EMG activity. The association between sympathetic arousal and muscle activity is of importance for understanding the high prevalence of musculoskeletal disorders in mentally stressful but physically light work tasks. Men had higher blood pressure and a more pronounced increase in epinephrine output than women, whereas women had higher heart rate. It was concluded that sympathetic activity is more sensitive to moderately intense stress exposure than pituitary adrenocortical (cortisol) activity and that men respond to performance stress with more epinephrine output than women. Although the correlations between the different indicators of sympathetic arousal were high, together they could still only explain 30-70% of the inter-individual variance. Thus, several parameters are needed in order to obtain a reliable measure of sympathetic activity. PMID- 15759599 TI - An evaluation of the ability to voluntarily reduce the heart rate after a month of yoga practice. AB - The study aimed at determining whether novices to yoga would be able to reduce their heart rate voluntarily and whether the magnitude of reduction would be more after 30 days of yoga training. Two groups (yoga and control, n = 12 each) were assessed on Day 1 and on Day 30. During the intervening 30 days, the yoga group received training in yoga techniques while the control group carried on with their routine. At each assessment the baseline heart rate was recorded for one minute, this was followed by a six-minute period during which participants were asked to attempt to voluntarily reduce their heart rate, using any strategy. Both the baseline heart rate and the lowest heart rate achieved voluntarily during the six-minute period were significantly lower in the yoga group on Day 30 compared to Day 1 by a group average of 10.7 beats per minute (i.e., bpm) and 6.8 bpm, respectively (p < .05, Wilcoxon paired signed ranks test). In contrast, there was no significant change in either the baseline heart rate or the lowest heart rate achieved voluntarily in the control group on Day 30 compared to Day 1. The results suggest that yoga training can enable practitioners to use their own strategies to reduce the heart rate, which has possible therapeutic applications. PMID- 15759600 TI - Effects of a carbohydrate supplement upon resting brain activity. AB - Glucose is a major energy source for the brain, and along with several monosaccharide derivatives as components of brain gangliosides, they play important roles in neurologic function. However, there is little information available on the role of glucose and other monosaccharides on resting brain activity. This study was designed to evaluate the effects of a single dose of a carbohydrate supplement containing glucose and several of its derivatives on resting brain activity in 20 healthy male college students. The supplement provided an insignificant amount of carbohydrate (3.9 g), protein (0.28 g), fat (0 g), and calories (14 kcal). The amount of glucose in the supplement was 0.5 g (1% the amount of glucose used in adult studies of cognitive functioning and memory). We hypothesized that the glyconutrient supplement would enhance brain activity associated with alertness and attention. The study design was double blind, with subjects randomly assigned to one of two orders, either carbohydrate supplement week one followed by placebo a week later, or the opposite. Electrical brain activity was monitored by 15 electrodes positioned at nine standard international 10-20 system locations, including three bilateral pairs at frontal, parietal, and occipital sites. Thirty minutes following ingestion of a placebo or carbohydrate supplement drink, EEG activity was recorded for 10-mins while subjects focused on a stationary visual target. Spectral power of resting brain activity was computed and analyzed contrasting the placebo and supplement groups. Relative to placebo, the carbohydrate supplement significantly enhanced power in three brain wave frequencies (theta, alpha, and beta) that are known to be associated with attention and arousal. Since changes were observed in the supplement but not placebo group, our study suggests that additional sugars in the glyconutritional supplement facilitate enhancement of brain electrical activity. Whether the apparent enhancement of arousal in baseline recordings is associated with improved task performance remains to be determined. PMID- 15759601 TI - Pavlovian George Windholz (1931-2002): an exemplar of scholarly "observation and observation" and a critical contributor to psychology, and hence to behavioral neuroscience. AB - Although most members of the Pavlovian Society properly focus their efforts on empirical research, the scholarly, critical conceptual contributions of some individuals are also relevant to progress in psychology and behavioral neuroscience. This paper discusses the contributions of the late George Windholz (often in collaboration with Peter Lamal) as: (a) a historian of Pavlov's life and work; (b) an analyst of priority issues in psychology as a science; (c) a refuter of myths perpetrated by psychology texts. These contributions provide an example of the scholarly form of the motto "observation and observation," where the data used to test hypotheses comprise original documents (often in languages other than English) examined by the historian's critical eye. PMID- 15759603 TI - Getting a grip on green. Working toward environmentally friendly construction and operations. PMID- 15759602 TI - Rehab hospital blends cultures. PMID- 15759604 TI - The right call. Selecting an appropriate nurse communication system. PMID- 15759605 TI - Great outdoors. Developing a master plan for exterior spaces. PMID- 15759606 TI - The front lines: NFPA's final fall meeting stresses emergency preparedness. PMID- 15759607 TI - Measured performance: advice on benchmarking housekeeping operations. PMID- 15759608 TI - Biogeometry: applications of computational geometry to molecular structure. PMID- 15759609 TI - Classification of non-coding RNA using graph representations of secondary structure. AB - Some genes produce transcripts that function directly in regulatory, catalytic, or structural roles in the cell. These non-coding RNAs are prevalent in all living organisms, and methods that aid the understanding of their functional roles are essential. RNA secondary structure, the pattern of base-pairing, contains the critical information for determining the three dimensional structure and function of the molecule. In this work we examine whether the basic geometric and topological properties of secondary structure are sufficient to distinguish between RNA families in a learning framework. First, we develop a labeled dual graph representation of RNA secondary structure by adding biologically meaningful labels to the dual graphs proposed by Gan et al [1]. Next, we define a similarity measure directly on the labeled dual graphs using the recently developed marginalized kernels [2]. Using this similarity measure, we were able to train Support Vector Machine classifiers to distinguish RNAs of known families from random RNAs with similar statistics. For 22 of the 25 families tested, the classifier achieved better than 70% accuracy, with much higher accuracy rates for some families. Training a set of classifiers to automatically assign family labels to RNAs using a one vs. all multi-class scheme also yielded encouraging results. From these initial learning experiments, we suggest that the labeled dual graph representation, together with kernel machine methods, has potential for use in automated analysis and classification of uncharacterized RNA molecules or efficient genome-wide screens for RNA molecules from existing families. PMID- 15759610 TI - An adaptive dynamic programming algorithm for the side chain placement problem. AB - Larger rotamer libraries, which provide a fine grained discretization of side chain conformation space by sampling near the canonical rotamers, allow protein designers to find better conformations, but slow down the algorithms that search for them. We present a dynamic programming solution to the side chain placement problem which treats rotamers at high or low resolution only as necessary. Dynamic programming is an exact technique; we turn it into an approximation, but can still analyze the error that can be introduced. We have used our algorithm to redesign the surface residues of ubiquitin's beta sheet. PMID- 15759611 TI - Computational design of combinatorial peptide library for modulating protein protein interactions. AB - Screening phage-displayed combinatorial peptide library is an effective approach for discovery of peptide modulators for protein-protein interactions. However, as peptide length increases, the chance of finding active peptides in a finite size library diminishes. To increase the likelihood of finding peptides that bind to a protein, we develop statistical potential for computational construction of biased combinatorial antibody-like peptide libraries. Based on the alpha shapes of antibody-antigen complexes, we developed an empirical pair potential for antigen-antibody interactions that depends on local packing. We validate this potential and show that it can successfully discriminate the native interface peptides from a simulated library of 10,000 random peptides for 34 antigen antibody complexes. In addition, we show that it can successfully recognize the native binding surface patch among all possible surface patches taken from either the antibody or the antigen for seven antibody-antigen protein complexes contained in the CAPRI (Critical Assessment of Predicted Interactions) dataset. We then develop a Weighted Amino Acid Residue sequence Generator (WAARG) for design of biased peptide library. When compared with a random peptide library, WAARG libraries contain more native-like binding peptides at a significantly smaller size. Our method can be used to construct peptide library for screening of antibody variants with improved specificity and affinity to a target antigen. It can also be used for screening of antibody-like antagonist peptides modulating other protein-protein interactions. PMID- 15759612 TI - Exploring protein folding trajectories using geometric spanners. AB - We describe the 3-D structure of a protein using geometric spanners--geometric graphs with a sparse set of edges where paths approximate the n2 inter-atom distances. The edges in the spanner pick out important proximities in the structure, labeling a small number of atom pairs or backbone region pairs as being of primary interest. Such compact multiresolution views of proximities in the protein can be quite valuable, allowing, for example, easy visualization of the conformation over the entire folding trajectory of a protein and segmentation of the trajectory. These visualizations allow one to easily detect formation of secondary and tertiary structures as the protein folds. PMID- 15759613 TI - Chaintweak: sampling from the neighbourhood of a protein conformation. AB - When searching for an optimal protein structure, it is often necessary to generate a set of structures similar, e.g., within 4A Root Mean Square Deviation (RMSD), to some base structure. Current methods to do this are designed to produce only small deviations (< 0.1A RMSD) and are inefficient for larger deviations. The method proposed in this paper, ChainTweak, can generate conformations with larger deviations from the base much more efficiently. For example, in 18 seconds it can generate 100 backbone conformations, each within 1 4A RMSD of a given 45-residue conformation. Moreover, each conformation has correct bond lengths, angles and omega torsional angles; its phi-psi angles have energetically favorable values; and there are rarely any backbone steric clashes. The method uses the insight that loop closure techniques can be used to perform compensatory changes of dihedral angles so that only a part of the conformation is changed. It is demonstrated, using decoys from the Decoys 'R Us data-set, that ChainTweak can be used to construct good decoys. It also provides a novel and intuitive way of analyzing the energy landscape of a protein. In addition, ChainTweak can improve the accuracy and performance of the loop modeling program RAPPER by an order of magnitude (1.1 min. vs. 36 min. for an 8-residue chain). PMID- 15759615 TI - Biomedical ontologies. PMID- 15759614 TI - Coarse and reliable geometric alignment for protein docking. AB - We present an efficient algorithm for generating a small set of coarse alignments between interacting proteins using meaningful features on their surfaces. The proteins are treated as rigid bodies, but the results are more generally useful as the produced configurations can serve as input to local improvement algorithms that allow for protein flexibility. We apply our algorithm to a diverse set of protein complexes from the Protein Data Bank, demonstrating the effectivity of our algorithm, both for bound and for unbound protein docking problems. PMID- 15759616 TI - Ontology driven dynamic linking of biology resources. AB - Biologists were early adopters of the Web and continue to use it as the primary means of delivering data, tools and knowledge to their community. The Web is made by the links between pages, yet these links have many limitations: they are static and maintained by hand; they can only link one lexical item to another single resource; ownership is necessary for the placement of link anchors and the link mechanism is essentially inflexible. Dynamic linking services, supported by ontologies, offer a mechanism to overcome such restrictions. The Conceptual Open Hypermedia Service (COHSE) system enhances web resources through the dynamic addition of hypertext links. These links are derived through the use of an ontology and associated lexicon along with a mapping from concepts to possible link targets. We describe an application of COHSE to Bioinformatics, using the Gene Ontology (GO) as an ontology and associated keyword mappings and GO associations as link targets. The resulting demonstrator (referred to here as GOHSE) provides both glossary functionality and the possibility of building knowledge based hypertext structures linking bioinformatics resources. PMID- 15759617 TI - Non-lexical approaches to identifying associative relations in the gene ontology. AB - The Gene Ontology (GO) is a controlled vocabulary widely used for the annotation of gene products. GO is organized in three hierarchies for molecular functions, cellular components, and biological processes but no relations are provided among terms across hierarchies. The objective of this study is to investigate three non lexical approaches to identifying such associative relations in GO and compare them among themselves and to lexical approaches. The three approaches are: computing similarity in a vector space model, statistical analysis of co occurrence of GO terms in annotation databases, and association rule mining. Five annotation databases (FlyBase, the Human subset of GOA, MGI, SGD, and WormBase) are used in this study. A total of 7,665 associations were identified by at least one of the three non-lexical approaches. Of these, 12% were identified by more than one approach. While there are almost 6,000 lexical relations among GO terms, only 203 associations were identified by both non-lexical and lexical approaches. The associations identified in this study could serve as the starting point for adding associative relations across hierarchies to GO, but would require manual curation. The application to quality assurance of annotation databases is also discussed. PMID- 15759618 TI - Genestrace: phenomic knowledge discovery via structured terminology. AB - The era of applied genomic medicine is quickly approaching accompanied by the increasing availability of detailed genetic information. Understanding the genetic etiology behind complex, multi-gene diseases remains an important challenge. In order to uncover the putative genetic etiology of complex diseases, we designed a method that explores the relationships between two major terminological and ontological resources: the Unified Medical Language System (UMLS) and the Gene Ontology (GO). The UMLS has a mainly clinical emphasis; Gene Ontology has become the standard for biological annotations of genes and gene products. Using statistical and semantic relationships within and between the two resources, we are able to infer relationships between disease concepts in the UMLS and gene products annotated using GO and its associated databases. We validated our inferences by comparing them to the known gene-disease relationships, as defined in the Online Mendelian Inheritance in Man's morbidmap (OMIM). The proof-of-concept methods presented here are unique in that they bypass the ambiguity of the direct extraction of gene or disease term from MEDLINE. Additionally, our methods provide direct links to clinically significant diseases through established terminologies or ontologies. The preliminary results presented here indicate the potential utility of exploiting the existing, manually curated relationships in biomedical resources as a tool for the discovery of potentially valuable new gene-disease relationships. PMID- 15759619 TI - Integrating genomic knowledge sources through an anatomy ontology. AB - Modern genomic research has access to a plethora of knowledge sources. Often, it is imperative that researchers combine and integrate knowledge from multiple perspectives. Although some technology exists for connecting data and knowledge bases, these methods are only just beginning to be successfully applied to research in modem cell biology. In this paper, we argue that one way to integrate multiple knowledge sources is through anatomy--both generic cellular anatomy, as well as anatomic knowledge about the tissues and organs that may be studied via microarray gene expression experiments. We present two examples where we have combined a large ontology of human anatomy (the FMA) with other genomic knowledge sources: the gene ontology (GO) and the mouse genomic databases (MGD) of the Jackson Labs. These two initial examples of knowledge integration provide a proof of concept that anatomy can act as a hub through which we can usefully combine a variety of genomic knowledge and data. PMID- 15759620 TI - Gotrees: predicting go associations from protein domain composition using decision trees. AB - The Gene Ontology (GO) offers a comprehensive and standardized way to describe a protein's biological role. Proteins are annotated with GO terms based on direct or indirect experimental evidence. Term assignments are also inferred from homology and literature mining. Regardless of the type of evidence used, GO assignments are manually curated or electronic. Unfortunately, manual curation cannot keep pace with the data, available from publications and various large experimental datasets. Automated literature-based annotation methods have been developed in order to speed up the annotation. However, they only apply to proteins that have been experimentally investigated or have close homologs with sufficient and consistent annotation. One of the homology-based electronic methods for GO annotation is provided by the InterPro database. The InterPro2GO/PFAM2GO associates individual protein domains with GO terms and thus can be used to annotate the less studied proteins. However, protein classification via a single functional domain demands stringency to avoid large number of false positives. This work broadens the basic approach. We model proteins via their entire functional domain content and train individual decision tree classifiers for each GO term using known protein assignments. We demonstrate that our approach is sensitive, specific and precise, as well as fairly robust to sparse data. We have found that our method is more sensitive when compared to the InterPro2GO performance and suffers only some precision decrease. In comparison to the InterPro2GO we have improved the sensitivity by 22%, 27% and 50% for Molecular Function, Biological Process and Cellular GO terms respectively. PMID- 15759621 TI - The clinical bioinformatics ontology: a curated semantic network utilizing RefSeq information. AB - Existing medical vocabularies lack rich terms to describe findings that are generated by modem molecular diagnostic procedures. Most bioinformatics resources were designed primarily to support the needs of the research community. We describe the development of a curated resource, the Clinical Bioinformatics Ontology (CBO), a semantic network appropriate for describing clinically significant genomics concepts. The CBO includes concepts appropriate for both molecular diagnostics and cytogenetics. A standardized methodology based on consistent application of RefSeq information is applied to the curation of the CBO in order to provide a reproducible and reliable tool. Challenges related to this curation process are discussed in this paper. At the time of submission the CBO included 4,069 concepts, associated by 8,463 relationships. PMID- 15759622 TI - Go molecular function terms are predictive of subcellular localization. AB - A protein's function is closely linked to its subcellular localization. Use of Gene Ontology (GO) molecular function terms to extend sequence-based subcellular localization prediction has been previously shown to improve predictive performance. Here, we explore directly the relationship between GO function annotations and localization information, identifying both highly predictive single terms, and terms with large information gain with respect to location. The results identify a number of predictive and informative GO terms with respect to subcellular location, particularly nucleus, extracellular space, membrane, mitochondrion, endoplasmic reticulum and Golgi. There are several clear examples illustrating why the addition of function information provides additional predictive power over sequence alone. Other interesting phenomena can also be seen in the results. Most predictive or informative terms are imperfect, and incorrect prediction may often call out significant biological phenomena. Finally, these results may be useful in the GO annotation process. PMID- 15759623 TI - Linking ontological resources using aggregatable substance identifiers to organize extracted relations. AB - Systems that extract biological regulatory pathway relations from free-text sources are intended to help researchers leverage vast and growing collections of research literature. Several systems to extract such relations have been developed but little work has focused on how those relations can be usefully organized (aggregated) to support visualization systems or analysis algorithms. Ontological resources that enumerate name strings for different types of biomedical objects should play a key role in the organization process. In this paper we delineate five potentially useful levels of relational granularity and propose the use of aggregatable substance identifiers to help reduce lexical ambiguity. An aggregatable substance identifier applies to a gene and its products. We merged 4 extensive lexicons and compared the extracted strings to the text of five million MEDLINE abstracts. We report on the ambiguity within and between name strings and common English words. Our results show an 89% reduction in ambiguity for the extracted human substance name strings when using an aggregatable substance approach. PMID- 15759624 TI - Implications of compositionality in the gene ontology for its curation and usage. AB - In this paper we argue that a richer underlying representational model for the Gene Ontology that captures the implicit compositional structure of GO terms could have a positive impact on two activities crucial to the success of GO: ontology curation and database annotation. We show that many of the new terms added to GO in a one-year span appear to be compositional variations of other terms. We found that 90.2% of the 3,652 new terms added between July 2003 and July 2004 exhibited characteristics of compositionality. We also examine annotations available from the GO Consortium website that are either manually curated or automatically generated. We found that 74.5% and 63.2% of GO terms are seldom, if ever, used in manual and automatic annotations, respectively. We show that there are features that tend to distinguish terms that are used from those that are not. In order to characterize the effect of compositionality on the combinatorial properties of GO, we employ finite state automata that represent sets of GO terms. This representational tool demonstrates how ontologies can grow very fast, and also shows that small conceptual changes can directly result in a large number of changes to the terminology. We argue that the curation and annotation findings we report are influenced by the combinatorial properties that present themselves in an ontology that does not have a model that properly captures the compositional structure of its terms. PMID- 15759625 TI - The use of common ontologies and controlled vocabularies to enable data exchange and deposition for complex proteomic experiments. AB - Controlled vocabularies provide a roadmap through complex biological data. Proteomic data is increasing in volume and is currently poorly served by public repositories due to the large number of different formats in which the data is generated and stored. The Human Proteome Organization Proteome Standards Initiative is establishing standards for data transfer and deposition. These standards utilize ontologies and controlled vocabularies to describe experimental procedures and common processes such as sample preparation This paper will discuss the development of such ontologies by the user community and their current utilization in the fields of protein:proein interactions and mass spectrometry. PMID- 15759626 TI - A flexible measure of contextual similarity for biomedical terms. AB - We present a measure of contextual similarity for biomedical terms. The contextual features need to be explored, because newly coined terms are not explicitly described and efficiently stored in biomedical ontologies and their inner features (e.g. morphologic or orthographic) do not always provide sufficient information about the properties of the underlying concepts. The context of each term can be represented as a sequence of syntactic elements annotated with biomedical information retrieved from an ontology. The sequences of contextual elements may be matched approximately by edit distance defined as the minimal cost incurred by the changes (including insertion, deletion and replacement) needed to transform one sequence into the other. Our approach augments the traditional concept of edit distance by elements of linguistic and biomedical knowledge, which together provide flexible selection of contextual features and their comparison. PMID- 15759627 TI - Understanding the global properties of functionally-related gene networks using the gene ontology. AB - The global behavior of interactions between genes can be investigated by forming the network of functionally-related genes using the annotations based on the Gene Ontology. We define two genes to be connected when the pair of genes is involved in the same biological process. There has been other work on the analysis of different kinds of cellular and metabolic networks, such as gene coexpression network, in which genes are paired when they are found to be coexpressed in the microarray experiments. We observe that our functionally-related gene networks among humans, fruit flies, worms and yeast exhibit the small-world property, but all except the network of worms show the existence of the scale-free property. PMID- 15759628 TI - Identification of functional modules in protein complexes via hyperclique pattern discovery. AB - Proteins usually do not act isolated in a cell but function within complicated cellular pathways, interacting with other proteins either in pairs or as components of larger complexes. While many protein complexes have been identified by large-scale experimental studies, due to a large number of false-positive interactions existing in current protein complexes 10, it is still difficult to obtain an accurate understanding of functional modules, which encompass groups of proteins involved in common elementary biological function. In this paper, we present a hyperclique pattern discovery approach for extracting functional modules (hyperclique patterns) from protein complexes. A hyperclique pattern is a type of association pattern containing proteins that are highly affiliated with each other. The analysis of hyperclique patterns shows that proteins within the same pattern tend to present in the protein complex together. Also, statistically significant annotations of proteins in a pattern using the Gene Ontology suggest that proteins within the same hyperclique pattern more likely perform the same function and participate in the same biological process. More interestingly, the 3-D structural view of proteins within a hyperclique pattern reveals that these proteins physically interactwith each other. In addition, we show that several hyperclique patterns corresponding to different functions can participate in the same protein complex as independent modules. Finally, we demonstrate that a hyperclique pattern can be involved in different complexes performing different higher-order biological functions, although the pattern corresponds to a specific elementary biological function. PMID- 15759629 TI - Multi-aspect gene relation analysis. AB - Recent progress in high-throughput screening technologies has led to the production of massive amounts data that we can use to understand biological systems. To interpret this data, biologists often need to analyze the characteristics of a set of genes by using Gene Ontology (GO) annotation. We are proposing a novel method for assisting such an analysis. Given a set of genes, the method automatically extracts several analyzing aspects in terms of a subset of genes that are attached to some related GO terms. It then creates a gene attribute bipartite graph that highlights the aspect selected by the user according to his/her interests. We describe this method in detail and report on an experiment where the proposed method is applied to the analysis of rat kidney expression data. PMID- 15759630 TI - Computational approaches for pharmacogenomics. PMID- 15759631 TI - Identification of genomic signatures for the design of assays for the detection and monitoring of anthrax threats. AB - Sequences that are present in a given species or strain while absent from or different in any other organisms can be used to distinguish the target organism from other related or un-related species. Such DNA signatures are particularly important for the identification of genetic source of drug resistance of a strain or for the detection of organisms that can be used as biological agents in warfare or terrorism. Most approaches used to find DNA signatures are laboratory based, require a great deal of effort and can only distinguish between two organisms at a time. We propose a more efficient and cost-effective bioinformatics approach that allows identification of genomic fingerprints for a target organism. We validated our approach using a custom microarray, using sequences identified as DNA fingerprints of Bacillus anthracis. Hybridization results showed that the sequences found using our algorithm were truly unique to B. anthracis and were able to distinguish B. anthracis from its close relatives B. cereus and B. thuringiensis. PMID- 15759632 TI - Enhancing data sharing in collaborative research projects with DASH. AB - We describe a software framework, called DASH, that enables the facile access, maintenance, curation and sharing of computational biology data among collaborating research scientists. The DASH event-based framework enables members of team-based research projects to describe the multistep computational processing pipelines frequently required to generate data for sharing, monitors multiple distributed data stores for changes, and will then automatically invoke the appropriate processing pipeline(s). These pipelines can be used to communicate the results of data analyses to collaborators using mechanisms such as Web Services. We describe the overall design of the DASH system and the application of a simple DASH prototype to a collaborative pharmacogenomics research project involving several dozen researchers located at several different sites--the UCSF Pharmacogenetics of Membrane Transporters project. PMID- 15759633 TI - Threshold gradient descent method for censored data regression with applications in pharmacogenomics. AB - An important area of research in pharmacogenomics is to relate high-dimensional genetic or genomic data to various clinical phenotypes of patients. Due to large variability in time to certain clinical event among patients, studying possibly censored survival phenotypes can be more informative than treating the phenotypes as categorical variables. In this paper, we develop a threshold gradient descent (TGD) method for the Cox model to select genes that are relevant to patients' survival and to build a predictive model for the risk of a future patient. The computational difficulty associated with the estimation in the high-dimensional and low-sample size settings can be efficiently solved by the gradient descent iterations. Results from application to real data set on predicting survival after chemotherapy for patients with diffuse large B-cell lymphoma demonstrate that the proposed method can be used for identifying important genes that are related to time to death due to cancer and for building a parsimonious model for predicting the survival of future patients. The TGD based Cox regression gives better predictive performance than the L2 penalized regression and can select more relevant genes than the L1 penalized regression. PMID- 15759634 TI - Parameterization of a nonlinear genotype to phenotype map using molecular networks. AB - Mathematical models of networks of molecular interactions controlling the expression of traits could theoretically be used as genotype to phenotype (GP) maps. Such maps are nonlinear functions of the environment and the genotype. It is possible to use nonlinear least square minimization methods to fit a model to a set of phenotypic data but the convergence of these methods is not automatic and may lead to a multiplicity of solutions. Both factors raise a number of questions with respect to using molecular networks as nonlinear maps. A method to fit a molecular network representing a bistable switch to various types of phenotypic data is introduced. This method relies on the identification of the model stable steady states and the estimation of the proportion of cells in each of them. By using environmental perturbations, it is possible to collect time series of phenotypic data resulting in a smooth objective function leading to a good estimate of the parameters used to generate the simulated phenotypes. PMID- 15759635 TI - Exploratory visual analysis of pharmacogenomic results. AB - Comprehensive analysis of expansive pharmacogenomic datasets is a daunting challenge. A thorough exploration of experimental results requires both statistical and annotative information. Therefore, appropriate analysis tools must bring a readily-accessible, flexible combination of statistics and biological annotation to the user's desktop. We present the Exploratory Visual Analysis (EVA) software and database as such a tool and demonstrate its utility in replicating the findings of an earlier pharmacogenomic study as well as elucidating novel biologically plausible hypotheses. EVA brings all of the often disparate pieces of analysis together in an infinitely flexible visual display that is amenable to any type of statistical result and biological question. Here, we describe the motivations for developing EVA, detail the database and custom graphical user interface (GUI), provide an example of its application to a publicly available pharmacogenomic dataset, and discuss the broad utility of the EVA tool for the pharmacogenomics community. PMID- 15759636 TI - Study of effect of drug lexicons on medication extraction from electronic medical records. AB - Extraction of relevant information from free-text clinical notes is becoming increasingly important in healthcare to provide personalized care to patients. The purpose of this dictionary-based NLP study was to determine the effects of using varying drug lexicons to automatically extract medication information from electronic medical records. A convenience training sample of 52 documents, each containing at least one medication, and a randomized test sample of 100 documents were used in this study. The training and test set documents contained a total of 681 and 641 medications respectively. Three sets of drug lexicons were used as sources for medication extraction: first, containing drug name and generic name; second with drug, generic and short names; third with drug, generic and short names followed by filtering techniques. Extraction with the first drug lexicon resulted in 83.7% sensitivity and 96.2% specificity for the training set and 85.2% sensitivity and 96.9% specificity for the test set. Adding the list of short names used for drugs resulted in increasing sensitivity to 95.0%, but decreased the specificity to 79.2% for the training set. Similar results of increased sensitivity of 96.4% and 80.1% specificity were obtained for the test set. Combination of a set of filtering techniques with data from the second lexicon increased the specificity to 98.5% and 98.8% for the training and test sets respectively while slightly decreasing the sensitivity to 94.1% (training) and 95.8% (test). Overall, the lexicon with filtering resulted in the highest precision, i.e., extracted the highest number of medications while keeping the number of extracted non-medications low. PMID- 15759637 TI - Introduction to informatics approaches in structural genomics: modeling and representation of function from macromolecular structure. AB - Despite the advantages provided by the enormous recent increases in the availability of structural information, functional assignment for the large number of proteins represented in the sequence and structural genomics projects remains a pressing problem for genomic era biology. This section describes work relevant to this problem from several perspectives, including new approaches that take advantage of combined structure and sequence-based classification. Leveraging of genomic context and evolutionary information to improve classification and predictive power is a second prominent theme in the papers represented here. Finally, issues in building a database for linking sequence, structural, and functional information are explored. PMID- 15759638 TI - Subfamily hmms in functional genomics. AB - The limitations of homology-based methods for prediction of protein molecular function are well known; differences in domain structure, gene duplication events and errors in existing database annotations complicate this process. In this paper we present a method to detect and model protein subfamilies, which can be used in high-throughput, genome-scale phylogenomic inference of protein function. We demonstrate the method on a set of nine PFAM families, and show that subfamily HMMs provide greater separation of homologs and non-homologs than is possible with a single HMM for each family. We also show that subfamily HMMs can be used for functional classification with a very low expected error rate. The BETE method for identifying functional subfamilies is illustrated on a set of serotonin receptors. PMID- 15759639 TI - Algorithms for structural comparison and statistical analysis of 3D protein motifs. AB - The comparison of structural subsites in proteins is increasingly relevant to the prediction of their biological function. To address this problem, we present the Match Augmentation algorithm (MA). Given a structural motif of interest, such as a functional site, MA searches a target protein structure for a match: the set of atoms with the greatest geometric and chemical similarity. MA is extremely efficient because it exploits the fact that the amino acids in a structural motif are not equally important to function. Using motif residues ranked on functional significance via the Evolutionary Trace (ET), MA prioritizes its search by initially forming matches with functionally significant residues, then, guided by ET, it augments this partial match stepwise until the whole motif is found. With this hierarchical strategy, MA runs considerably faster than other methods, and almost always identifies matches in homologs known to have cognate functional sites. Second, in order to interpret matches, we further introduce a statistical method using nonparametric density estimation of the frequency distribution of structural matches. Our results show that the hierarchy of functional importance within structural motifs speeds up the search within targets, and points to a new method to score their statistical significance. PMID- 15759640 TI - Two-stage multi-class support vector machines to protein secondary structure prediction. AB - Bioinformatics techniques to protein secondary structure (PSS) prediction are mostly single-stage approaches in the sense that they predict secondary structures of proteins by taking into account only the contextual information in amino acid sequences. In this paper, we propose two-stage Multi-class Support Vector Machine (MSVM) approach where a MSVM predictor is introduced to the output of the first stage MSVM to capture the sequential relationship among secondary structure elements for the prediction. By using position specific scoring matrices, generated by PSI-BLAST, the two-stage MSVM approach achieves Q3 accuracies of 78.0% and 76.3% on the RS126 dataset of 126 nonhomologous globular proteins and the CB396 dataset of 396 nonhomologous proteins, respectively, which are better than the highest scores published on both datasets to date. PMID- 15759641 TI - Representing structure-function relationships in mechanistically diverse enzyme superfamilies. AB - The prediction of protein function from structure or sequence data remains a problem best addressed by leveraging information available from previously determined structure-function relationships. In the case of enzymes, the study of mechanistically diverse superfamilies can provide a rich source of structure function information useful in functional determination and enzyme engineering. To access these relationships using a computational resource, several issues must be addressed regarding the representation of enzyme function, the organization of structure-function relationships in the superfamily context, the handling of misannotations, and reliability of classifications and evidence. We discuss here our approaches to solving these problems in the development of a Structure Function Linkage Database (SFLD) (online at http://sfld.rbvi.ucsf.edu). PMID- 15759642 TI - Discovering sequence-structure motifs from protein segments and two applications. AB - We present a novel method for clustering short protein segments having strong sequence-structure correlations, and demonstrate that these clusters contain useful structural information via two applications. When applied to local tertiary structure prediction, we achieve approximately 60% accuracy with a novel dynamic programming algorithm. When applied to secondary structure prediction based on Support Vector Machines, we obtain a approximately 2% gain in Q3 performance by incorporating cluster-derived data into training and classification. These encouraging results illustrate the great potential of using conserved local motifs to tackle protein structure predictions and possibly other important problems in biology. PMID- 15759643 TI - Inferring SNP function using evolutionary, structural, and computational methods. PMID- 15759644 TI - Fast and cheap genome wide haplotype construction via optical mapping. AB - We describe an efficient algorithm to construct genome wide haplotype restriction maps of an individual by aligning single molecule DNA fragments collected with Optical Mapping technology. Using this algorithm and small amount of genomic material, we can construct the parental haplotypes for each diploid chromosome for any individual. Since such haplotype maps reveal the polymorphisms due to single nucleotide differences (SNPs) and small insertions and deletions (RFLPs), they are useful in association studies, studies involving genomic instabilities in cancer, and genetics, and yet incur relatively low cost and provide high throughput. If the underlying problem is formulated as a combinatorial optimization problem, it can be shown to be NP-complete (a special case of K population problem). But by effectively exploiting the structure of the underlying error processes and using a novel analog of the Baum-Welch algorithm for HMM models, we devise a probabilistic algorithm with a time complexity that is linear in the number of markers for an epsilon-approximate solution. The algorithms were tested by constructing the first genome wide haplotype restriction map of the microbe T. pseudoana, as well as constructing a haplotype restriction map of a 120 Mb region of Human chromosome 4. The frequency of false positives and false negatives was estimated using simulated data. The empirical results were found very promising. PMID- 15759645 TI - Improving functional annotation of non-synonomous SNPs with information theory. AB - Automated functional annotation of nsSNPs requires that amino-acid residue changes are represented by a set of descriptive features, such as evolutionary conservation, side-chain volume change, effect on ligand-binding, and residue structural rigidity. Identifying the most informative combinations of features is critical to the success of a computational prediction method. We rank 32 features according to their mutual information with functional effects of amino-acid substitutions, as measured by in vivo assays. In addition, we use a greedy algorithm to identify a subset of highly informative features. The method is simple to implement and provides a quantitative measure for selecting the best predictive features given a set of features that a human expert believes to be informative. We demonstrate the usefulness of the selected highly informative features by cross-validated tests of a computational classifier, a support vector machine (SVM). The SVM's classification accuracy is highly correlated with the ranking of the input features by their mutual information. Two features describing the solvent accessibility of "wild-type" and "mutant" amino-acid residues and one evolutionary feature based on superfamily-level multiple alignments produce comparable overall accuracy and 6% fewer false positives than a 32-feature set that considers physiochemical properties of amino acids, protein electrostatics, amino-acid residue flexibility, and binding interactions. PMID- 15759646 TI - From context-dependence of mutations to molecular mechanisms of mutagenesis. AB - Mutation frequencies vary significantly along nucleotide sequences such that mutations often concentrate at certain positions called hotspots. Mutation hotspots in DNA reflect intrinsic properties of the mutation process, such as sequence specificity, that manifests itself at the level of interaction between mutagens, DNA, and the action of the repair and replication machineries. The nucleotide sequence context of mutational hotspots is a fingerprint of interactions between DNA and repair/replication/modification enzymes, and the analysis of hotspot context provides evidence of such interactions. The hotspots might also reflect structural and functional features of the respective DNA sequences and provide information about natural selection. We discuss analysis of 8-oxoguanine-induced mutations in pro- and eukaryotic genes, polymorphic positions in the human mitochondrial DNA and mutations in the HIV-1 retrovirus. Comparative analysis of 8-oxoguanine-induced mutations and spontaneous mutation spectra suggested that a substantial fraction of spontaneous A x T-->C x T mutations is caused by 8-oxoGTP in nucleotide pools. In the case of human mitochondrial DNA, significant differences between molecular mechanisms of mutations in hypervariable segments and coding part of DNA were detected. Analysis of mutations in the HIV-1 retrovirus suggested a complex interplay between molecular mechanisms of mutagenesis and natural selection. PMID- 15759647 TI - A Bayesian framework for SNP identification. AB - As evolutionary models for single-nucleotide polymorphisms (SNPs) become available, methods for using them in the context of evolutionary information and expert prior information is a necessity. We formulate a probability model for SNPs as a Bayesian inference problem. Using this framework we compare the individual and combined predictive ability of four evolutionary models of varying levels of specificity on three SNP databases (two specifically targeted at functional SNPs) by calculating posterior probabilities and generating Receiver Operating Characteristic (ROC) curves. We discover that none of the models do exceptionally well, in some cases no better than a random-guess model. However, we demonstrate that several properties of the Bayesian formulation improve the predictability of SNPs in the three databases, specifically the ability to utilize mixtures of evolutionary models and a prior based on the genetic code. PMID- 15759649 TI - Joint learning from multiple types of genomic data. PMID- 15759648 TI - Untangling the effects of codon mutation and amino acid exchangeability. AB - Determining the relative contributions of mutation and selection to evolutionary change is a matter of great practical and theoretical significance. In this paper, we examine relative contributions of codon mutation rates and amino acid exchangeability on the frequencies of each type of amino acid difference in alignments of distantly related proteins, alignments of closely related proteins, and among human SNPs, using a model that incorporates prior estimates of mutation and exchangeability parameters. For the operational exchangeability of amino acids in proteins, we use EX, a measure of protein-level effects from a recent statistical meta-analysis of nearly 10,000 experimental amino acid exchanges. EX is both free of mutational effects and more powerful than commonly used "biochemical distance" measures (1). For distant protein relationships, mutational effects (genetic code, transition/transversion bias) and operational exchangeability (EX) account for roughly equal portions of variance in off diagonal values, the complete model accounting for R2 = 0.35 of the variance. For human/chimpanzee alignments representing closely related proteins relationships, mutational effects (including CpG bias) account for 0.52 of the variance; adding EX to the model increases this to 0.67. For natural variation in human proteins, the variance explained by mutational effects alone, and by mutational effects and operational exchangeability are, respectively, 0.66 and 0.70 for SNPs in HGVBase, and 0.56 and 0.60 for disease-causing missense variants in HGMD. Thus, exchangeability has a stronger relative effect for distant protein evolution than for the cases of closely related proteins or of population variation. A more detailed model for the hominid data suggests that 1) there is a threshold in EX below which substitutions are highly unlikely to be accepted, corresponding to roughly 30 % relative protein activity; 2) selection against missense mutants is a slightly convex function of protein activity, not changing much as long as protein activity is low; and 3) the probability of disease-causing effects decreases nearly linearly with EX. PMID- 15759650 TI - Sparse factorizations of gene expression data guided by binding data. AB - Existing clustering methods do not deal well with overlapping clusters, are unstable and do not take into account the robustness of biological systems, or more complex background knowledge such as regulator binding data. Here we describe a nonnegative sparse factorization algorithm dealing with the above problems: cluster overlaps are allowed by design, the nonnegativity constraints implicitly approximate the robustness of biological systems and regulator binding data is used to guide the factorization. Preliminary results show the feasibility of our approach. PMID- 15759652 TI - Genome-scale protein function prediction in yeast Saccharomyces cerevisiae through integrating multiple sources of high-throughput data. AB - As we are moving into the post genome-sequencing era, various high-throughput experimental techniques have been developed to characterize biological systems at the genome scale. Discovering new biological knowledge from high-throughput biological data is a major challenge for bioinformatics today. To address this challenge, we developed a Bayesian statistical method together with Boltzmann machine and simulated annealing for protein function prediction in the yeast Saccharomyces cerevisiae through integrating various high-throughput biological data, including protein binary interactions, protein complexes and microarray gene expression profiles. In our approach, we quantified the relationship between functional similarity and high-throughput data. Based on our method, 1802 out of 2280 unannotated proteins in the yeast were assigned functions systematically. The related computer package is available upon request. PMID- 15759651 TI - Informative structure priors: joint learning of dynamic regulatory networks from multiple types of data. AB - We present a method for jointly learning dynamic models of transcriptional regulatory networks from gene expression data and transcription factor binding location data. Models are automatically learned using dynamic Bayesian network inference algorithms; joint learning is accomplished by incorporating evidence from gene expression data through the likelihood, and from transcription factor binding location data through the prior. We propose a new informative structure prior with two advantages. First, the prior incorporates evidence from location data probabilistically, allowing it to be weighed against evidence from expression data. Second, the prior takes on a factorable form that is computationally efficient when learning dynamic regulatory networks. Results obtained from both simulated and experimental data from the yeast cell cycle demonstrate that this joint learning algorithm can recover dynamic regulatory networks from multiple types of data that are more accurate than those recovered from each type of data in isolation. PMID- 15759653 TI - Discovering transcriptional modules from motif, chip-chip and microarray data. AB - We present a method for inference of transcriptional modules from heterogeneous data sources. It allows identifying the responsible set of regulators in combination with their corresponding DNA recognition sites (motifs) and target genes. Our approach distinguishes itself from previous work in literature because it fully exploits the knowledge of three independently acquired data sources: ChIP-chip data; motif information as obtained by phylogenetic shadowing; and gene expression profiles obtained using microarray experiments. Moreover, these three data sources are dealt with in a new and fully integrated manner. By avoiding approaches that take the different data sources into account sequentially or iteratively, the transparency of the method and the interpretability of the results are ensured. Using our method on biological data demonstrated the biological relevance of the inference. PMID- 15759654 TI - Genrate: a generative model that finds and scores new genes and exons in genomic microarray data. AB - Recently, researchers have made some progress in using microarrays to validate predicted exons in genome sequence and find new gene structures. However, current methods rely on separately making threshold-based decisions on intensity of expression, similarity of expression profiles, and arrangements of exons in the genome. We have taken a Bayesian approach and developed GenRate, a generative model that accounts for both genome-wide expression data taken from multiple conditions (e.g. tissues) and co-location and density of probes in DNA sequence data. GenRate balances probabilistic evidence derived from different sources and outputs scores (log-likelihoods) for each gene model, enabling the estimation of false-positive and false-negative rates. The model has a number of local minima that is exponential in the length of the DNA sequence data, so direct application of the EM learning algorithm produces poor results. We describe a novel way of parameterizing the model using examples from the data set, so that good solutions are found using an efficient algorithm. We apply GenRate to a subset of mouse genome-wide expression data that we have created, and discuss the statistical significance of the genes found by GenRate. Three of the highest-ranking gene structures found by GenRate, each containing thousands of bases from the genome, are confirmed using RT-PCR experiments. PMID- 15759655 TI - Bayesian joint prediction of associated transcription factors in Bacillus subtilis. AB - Sigma factors, often in conjunction with other transcription factors, regulate gene expression in prokaryotes at the transcriptional level. Specific transcription factors tend to co-occur with specific sigma factors. To predict new members of the transcription factor regulon, we applied Bayes rule to combine the Bayesian probability of sigma factor prediction calculated from microarray data and the sigma factor binding sequence motif, the motif score of the transcription factor associated with the sigma factor, the empirically determined distance between the transcription start site to the cis-regulatory region, and the tendency for specific sigma factors and transcription factors to co-occur. By combining these information sources, we improve the accuracy of predicting regulation by transcription factors, and also confirm the sigma factor prediction. We applied our proposed method to all genes in Bacillus subtilis to find currently unknown gene regulations by transcription factors and sigma factors. PMID- 15759657 TI - Random forest similarity for protein-protein interaction prediction from multiple sources. AB - One of the most important, but often ignored, parts of any clustering and classification algorithm is the computation of the similarity matrix. This is especially important when integrating high throughput biological data sources because of the high noise rates and the many missing values. In this paper we present a new method to compute such similarities for the task of classifying pairs of proteins as interacting or not. Our method uses direct and indirect information about interaction pairs to constructs a random forest (a collection of decision tress) from a training set. The resulting forest is used to determine the similarity between protein pairs and this similarity is used by a classification algorithm (a modified kNN) to classify protein pairs. Testing the algorithm on yeast data indicates that it is able to improve coverage to 20% of interacting pairs with a false positive rate of 50%. These results compare favorably with all previously suggested methods for this task indicating the importance of robust similarity estimates. PMID- 15759656 TI - Modulefinder: a tool for computational discovery of cis regulatory modules. AB - Regulation of gene expression occurs largely through the binding of sequence specific transcription factors (TFs) to genomic binding sites (BSs). We present a rigorous scoring scheme, implemented as a C program termed "ModuleFinder", that evaluates the likelihood that a given genomic region is a cis regulatory module (CRM) for an input set of TFs according to its degree of: (1) homotypic site clustering; (2) heterotypic site clustering; and (3) evolutionary conservation across multiple genomes. Importantly, ModuleFinder obtains all parameters needed to appropriately weight the relative contributions of these sequence features directly from the input sequences and TFBS motifs, and does not need to first be trained. Using two previously described collections of experimentally verified CRMs in mammals and in fly as validation datasets, we show that ModuleFinder is able to identify CRMs with great sensitivity and specificity. PMID- 15759658 TI - Updating software? How you can overcome islands of automation. PMID- 15759659 TI - Giving culturally competent care another element in patient safety. PMID- 15759661 TI - How customers rate surgical software. PMID- 15759660 TI - Novation splits up sutures, endo devices. PMID- 15759662 TI - Automation eases the preop process. PMID- 15759663 TI - Online patient histories help reduce need for phone tag. PMID- 15759664 TI - What's next for preop automation? PMID- 15759665 TI - Meeting patients' language needs. PMID- 15759666 TI - Getting a handle on scope repair costs. PMID- 15759667 TI - Striving for efficiency in cataract surgery. PMID- 15759669 TI - Some comments on Sabine absorption coefficient. PMID- 15759668 TI - Help! There are 60 screaming kids in my lab!--Outreach activities for 5th graders. PMID- 15759670 TI - A note on pure-tone masking by broadband noise under free-field and insert-phone conditions. PMID- 15759671 TI - Acoustical wave propagator for time-domain dynamic stress concentration in a plate with a sharp change of section. AB - The acoustical wave propagator technique is applied to study the time-domain dynamic stress concentration in a two-dimensional flexible plate with a sharp change of section. As a wave packet approaches the plate discontinuity where the sharp change of thickness is introduced, the spatial interference patterns in the displacement of the plate and internal stresses vary with time. The constructive interference of stresses is referred to as time-domain stress concentration. The superposition of wave fronts of incident and reflected wave packets is used to explain the spatial distribution of the interference patterns. The increase of dynamic stress near the vicinity of the discontinuity boundary of the plate is studied as a function of time and the thickness ratio of the plates. PMID- 15759672 TI - Equations for finite-difference, time-domain simulation of sound propagation in moving inhomogeneous media and numerical implementation. AB - Finite-difference, time-domain (FDTD) calculations are typically performed with partial differential equations that are first order in time. Equation sets appropriate for FDTD calculations in a moving inhomogeneous medium (with an emphasis on the atmosphere) are derived and discussed in this paper. Two candidate equation sets, both derived from linearized equations of fluid dynamics, are proposed. The first, which contains three coupled equations for the sound pressure, vector acoustic velocity, and acoustic density, is obtained without any approximations. The second, which contains two coupled equations for the sound pressure and vector acoustic velocity, is derived by ignoring terms proportional to the divergence of the medium velocity and the gradient of the ambient pressure. It is shown that the second set has the same or a wider range of applicability than equations for the sound pressure that have been previously used for analytical and numerical studies of sound propagation in a moving atmosphere. Practical FDTD implementation of the second set of equations is discussed. Results show good agreement with theoretical predictions of the sound pressure due to a point monochromatic source in a uniform, high Mach number flow and with Fast Field Program calculations of sound propagation in a stratified moving atmosphere. PMID- 15759673 TI - Approximate high-order dynamic theory of a fluid layer in between two thick solids. AB - A thin layer of heavy fluid with bound faces is considered first. For the sake of simplicity, the effects of vortex and viscosity are neglected. Two cases of compressible and incompressible fluids are treated, assuming that a thickness over-wavelength ratio is a small parameter. For the internal state of medium the relations between the quantities on the layer surfaces are deduced. In contrast to the standard approach, which expands the propagator matrix into power series of a small wave number, the asymptotic integration of 3D equations and boundary conditions of fluid dynamics is performed. Respective relations are represented in a recurrent form and permit one to obtain the high-order components of the displacements and pressure rather simply. When considering two thick solids with a fluid in between, this result is used to derive the so-called "impedance boundary conditions" (IBC) with a relative asymptotic error up to tenth order. Tests show their validity until the first quasiresonance frequency of a layer, so, the analysis is not just long wave but essentially low frequency. Thus, these IBC are applicable to reduce the dimension in the analysis of challenged multicomponent system of fluid-coupled solids in a reasonable frequency range. PMID- 15759674 TI - The attenuation of the higher-order cross-section modes in a duct with a thin porous layer. AB - A numerical method for sound propagation of higher-order cross-sectional modes in a duct of arbitrary cross-section and boundary conditions with nonzero, complex acoustic admittance has been considered. This method assumes that the cross section of the duct is uniform and that the duct is of a considerable length so that the longitudinal modes can be neglected. The problem is reduced to a two dimensional (2D) finite element (FE) solution, from which a set of cross sectional eigen-values and eigen-functions are determined. This result is used to obtain the modal frequencies, velocities and the attenuation coefficients. The 2D FE solution is then extended to three-dimensional via the normal mode decomposition technique. The numerical solution is validated against experimental data for sound propagation in a pipe with inner walls partially covered by coarse sand or granulated rubber. The values of the eigen-frequencies calculated from the proposed numerical model are validated against those predicted by the standard analytical solution for both a circular and rectangular pipe with rigid walls. It is shown that the considered numerical method is useful for predicting the sound pressure distribution, attenuation, and eigen-frequencies in a duct with acoustically nonrigid boundary conditions. The purpose of this work is to pave the way for the development of an efficient inverse problem solution for the remote characterization of the acoustic boundary conditions in natural and artificial waveguides. PMID- 15759675 TI - Measurements and empirical model of the acoustic properties of reticulated vitreous carbon. AB - Reticulated vitreous carbon (RVC) is a highly porous, rigid, open cell carbon foam structure with a high melting point, good chemical inertness, and low bulk thermal conductivity. For the proper design of acoustic devices such as acoustic absorbers and thermoacoustic stacks and regenerators utilizing RVC, the acoustic properties of RVC must be known. From knowledge of the complex characteristic impedance and wave number most other acoustic properties can be computed. In this investigation, the four-microphone transfer matrix measurement method is used to measure the complex characteristic impedance and wave number for 60 to 300 pore per-inch RVC foams with flow resistivities from 1759 to 10,782 Pa s m(-2) in the frequency range of 330 Hz-2 kHz. The data are found to be poorly predicted by the fibrous material empirical model developed by Delany and Bazley, the open cell plastic foam empirical model developed by Qunli, or the Johnson-Allard microstructural model. A new empirical power law model is developed and is shown to provide good predictions of the acoustic properties over the frequency range of measurement. Uncertainty estimates for the constants of the model are also computed. PMID- 15759676 TI - Investigation of the phase velocities of guided acoustic waves in soft porous layers. AB - A new experimental method for measuring the phase velocities of guided acoustic waves in soft poroelastic or poroviscoelastic plates is proposed. The method is based on the generation of standing waves in the material and on the spatial Fourier transform of the displacement profile of the upper surface. The plate is glued on a rigid substrate so that it has a free upper surface and a nonmoving lower surface. The displacement is measured with a laser Doppler vibrometer along a line corresponding to the direction of propagation of plane surface waves. A continuous sine with varying frequencies was chosen as excitation signal to maximize the precision of the measurements. The spatial Fourier transform provides the wave numbers, and the phase velocities are obtained from the relationship between wave number and frequency. The phase velocities of several guided modes could be measured in a highly porous foam saturated by air. The modes were also studied theoretically and, from the theoretical results, the experimental results, and a fitting procedure, it was possible to determine the frequency behavior of the complex shear modulus and of the complex Poisson ratio from 200 Hz to 1.4 kHz, in a frequency range higher than the traditional methods. PMID- 15759677 TI - Reconstruction of vibroacoustic fields in half-space by using hybrid near-field acoustical holography. AB - In this paper we examine the accuracy and efficiency of reconstructing the vibroacoustic quantities generated by a vibrating structure in half-space by using hybrid near-field acoustic holography (NAH) and modified Helmholtz equation least squares (HELS) formulations. In hybrid NAH, we combine modified HELS with an inverse boundary element method (IBEM) to reconstruct a vibroacoustic field. The main advantage of this approach is that the majority of the input data can be regenerated but not measured, thus the efficiency is greatly enhanced. In modified HELS, we expand the field acoustic pressure in terms of outgoing and incoming spherical waves and specify the corresponding expansion coefficients by solving a system of equations obtained by matching the assumed-form solution to the measured acoustic pressure. Here the system of equations is ill conditioned and Tikhonov regularization is implemented through singular value decomposition (SVD) and the generalized cross-validation (GCV) method. Numerical examples of a dilating and oscillating spheres and finite cylinder are demonstrated. Test results show that hybrid NAH can yield a more accurate reconstruction than does a modified HELS, but a modified HELS is more efficient than is hybrid NAH [Work supported by NSF]. PMID- 15759678 TI - Nonlinear focusing of acoustic shock waves at a caustic cusp. AB - The present study investigates the focusing of acoustical weak shock waves incoming on a cusped caustic. The theoretical model is based on the Khokhlov Zabolotskaya equation and its specific boundary conditions. Based on the so called Guiraud's similitude law for a step shock, a new explanation about the wavefront unfolding due to nonlinear self-refraction is proposed. This effect is shown to be associated not only to nonlinearities, as expected by previous authors, but also to the nonlocal geometry of the wavefront. Numerical simulations confirm the sensitivity of the process to wavefront geometry. Theoretical modeling and numerical simulations are substantiated by an original experiment. This one is carried out in two steps. First, the canonical Pearcey function is synthesized in linear regime by the inverse filter technique. In the second step, the same wavefront is emitted but with a high amplitude to generate shock waves during the propagation. The experimental results are compared with remarkable agreement to the numerical ones. Finally, applications to sonic boom are briefly discussed. PMID- 15759679 TI - High-intensity rocket noise: nonlinear propagation, atmospheric absorption, and characterization. AB - Analyses of rocket noise data measured at far-field locations during the launch of a large rocket and a smaller rocket are presented. Weak shocks are present in all of the data sets. In order to characterize these shocks, those segments of the waveforms where the acoustic pressure is increasing are isolated and the rate of increase in pressure plotted versus magnitude of pressure rise. The plots follow a trend consistent with random noise at low values of pressure rise, then transition to the pressure-squared dependence expected for weak shocks at higher pressure rise values. Power spectral densities of the noise during the period of maximum overall sound-pressure levels display high- and low-frequency spectral slopes that are close to those predicted for shock-dominated noise. It is concluded that shocks must be included in propagation models if high frequency levels are to be estimated as a function of distance from the source. Initial shock thicknesses will have to be characterized experimentally and will require instrumentation with a bandwidth well in excess of 20 kHz. Reflection-free data are essential if meaningful assessments of the statistical properties of the noise are to be made. PMID- 15759680 TI - Nonlinear acoustic scattering by a partially closed surface-breaking crack. AB - A theoretical model describing the nonlinear scattering of acoustic waves by surface-breaking cracks with faces in partial contact is presented. The nonlinear properties of the crack are accounted for by suitable boundary conditions that are derived from micromechanical models of the dynamics of elastic rough surfaces in contact. Both linear and nonlinear responses of the crack are shown to be largest for a shear vertical wave incident on the surface containing the crack at an angle just above the critical angle for longitudinal waves. These findings question the fitness for the purpose of a conventional inspection method, which utilizes shear vertical waves at 45 degrees of incidence to search for surface breaking cracks in many engineering components. For angles of incidence proximal to the critical angle of longitudinal waves, the efficiency of the second harmonic's generation appears to be the highest. Thanks to the increased sensitivity to surface-breaking cracks, this configuration seems to offer a solution to the localization problem, a task that has eluded nonlinear techniques operating under other circumstances. Finally, this model suggests a simple interpretation of the highly localized nonlinear response of delaminations in composite materials. PMID- 15759681 TI - Resonant properties of a nonlinear dissipative layer excited by a vibrating boundary: Q-factor and frequency response. AB - Simplified nonlinear evolution equations describing non-steady-state forced vibrations in an acoustic resonator having one closed end and the other end periodically oscillating are derived. An approach based on a nonlinear functional equation is used. The nonlinear Q-factor and the nonlinear frequency response of the resonator are calculated for steady-state oscillations of both inviscid and dissipative media. The general expression for the mean intensity of the acoustic wave in terms of the characteristic value of a Mathieu function is derived. The process of development of a standing wave is described analytically on the base of exact nonlinear solutions for different laws of periodic motion of the wall. For harmonic excitation the wave profiles are described by Mathieu functions, and their mean energy characteristics by the corresponding eigenvalues. The sawtooth shaped motion of the boundary leads to a similar process of evolution of the profile, but the solution has a very simple form. Some possibilities to enhance the Q-factor of a nonlinear system by suppression of nonlinear energy losses are discussed. PMID- 15759682 TI - Measurement and modeling of three-dimensional sound intensity variations due to shallow-water internal waves. AB - Broadband acoustic data (30-160 Hz) from the SWARM'95 experiment are analyzed to investigate acoustic signal variability in the presence of ocean internal waves. Temporal variations in the intensity of the received signals were observed over periods of 10 to 15 min. These fluctuations are synchronous in depth and are dependent upon the water column variability. They can be explained by significant horizontal refraction taking place when the orientation of the acoustic track is nearly parallel to the fronts of the internal waves. Analyses based on the equations of vertical modes and horizontal rays and on a parabolic equation in the horizontal plane are carried out and show interesting frequency-dependent behavior of the intensity. Good agreement is obtained between theoretical calculations and experimental data. PMID- 15759683 TI - Geoacoustic inversion with ships as sources. AB - Estimation of geoacoustic parameters using acoustic data from a surface ship was performed for a shallow water region in the Gulf of Mexico. The data were recorded from hydrophones in a bottom mounted, horizontal line array (HLA). The techniques developed to produce the geoacoustic inversion are described, and an efficient method for geoacoustic inversion with broadband beam cross-spectral data is demonstrated. The performance of cost functions that involve coherent or incoherent sums over frequency and one or multiple time segments is discussed. Successful inversions for the first sediment layer sound speed and thickness and some of the parameters for the deeper layers were obtained with the surface ship at nominal ranges of 20, 30, or 50 water depths. The data for these inversions were beam cross-spectra from four subapertures of the HLA spanning a little more than two water depths. The subaperture beams included ten frequencies equally spaced in the 120-200 Hz band. The values of the geoacoustic parameters from the inversions are validated by comparisons with geophysical observations and with the parameter values from previous inversions by other invesigators, and by comparing transmission loss (TL) measured in the experiment with modeled TL based on the inverted geoacoustic parameters. PMID- 15759684 TI - The reflection of ultrasound from partially contacting rough surfaces. AB - Ultrasound is commonly used to detect and size cracks in a range of engineering components. Modeling techniques are well established for smooth and open cracks. However, real cracks are often rough (relative to the ultrasonic wavelength) and closed due to compressive stress. This paper describes an investigation into the combined effects of crack face roughness and closure on ultrasonic detectability. A contact model has been used to estimate the size and shape of scatterers (voids) at the interface of these rough surfaces when loaded. The response of such interfaces to excitation with a longitudinal ultrasonic pulse over a wide range of frequencies has been investigated. The interaction of ultrasound with this scattering interface is predicted using a finite-element model and good agreement with experiments on rough surfaces is shown. Results are shown for arrays of equi-sized scatterers and a distribution of scatterer sizes. It is shown that the response at high frequencies is dependent on the size, shape, and distribution of the scatterers. It is also shown that the finite-element results depart from the mass-spring model predictions when the product of wave number and scatterer half-width is greater than 0.4. PMID- 15759685 TI - System for determination of ultrasonic wave speeds and their temperature dependence in liquids and in vitro tissues. AB - An interferometric technique capable of accurately measuring wave speed in liquids is reported. The hardware is adapted from a design to measure nonlinear responses of biological tissues to pressure changes (pressure derivatives) and temperature changes (temperature derivatives). It is used with the highly sensitive variable frequency pulsed phase-locked loop (VFPPLL) instrument. The system uses well-understood and well-characterized components and systems. The apparatus covers a temperature range from below 5 degrees C to above 45 degrees C. The system with the high-sensitivity VFPPLL is capable of measurement of wave speed to an uncertainty of less than 0.1%, and changes in wave speed to better than 0.001%. The transducer is an undamped temperature-characterized PZT-5A 500 kHz plate, whose output is corrected for off-resonance operation and for diffraction effects. To test the accuracy of the technique, measurement of ultrasonic compressional wave speed in water at temperatures from 10 degrees C to 45 degrees C are reported, with an estimated uncertainty of 0.07% and a temperature uncertainty of 0.15 degrees C. The agreement between mean values and literature values is better than 0.05%. PMID- 15759686 TI - Analysis of the sandwich piezoelectric ultrasonic transducer in coupled vibration. AB - The coupled vibration of the sandwich piezoelectric transducer with a large cross section is analyzed using an approximate analytic method. The resonance frequency equations of the transducer are derived and the effect of the geometrical dimensions on the resonance frequency is studied. It is illustrated that when the radial vibration in the transducer is considered, the vibration of the sandwich transducer becomes more complex. Apart from the longitudinal resonance frequency, the radial resonance frequency can also be obtained. For comparison, numerical methods are also used to simulate the coupled vibration; the resonance frequency and the vibrational displacement distribution are computed. Compared with one dimensional longitudinal theory, the radial dimensions of the transducer are no longer limited because the coupled vibration is considered. Compared with numerical methods, the physical meaning of the analytic method is concise. It is illustrated that the resonance frequencies obtained from the coupled resonance frequency equations are in good agreement with those from numerical methods, and they are in better agreement with the measured results than those from one dimensional theory. Since the radial and the coupled vibration are considered in the analysis, more resonance frequencies can be obtained. Therefore, using the coupled resonance frequency equations, the sandwich transducer with multifrequency or wide frequency bandwidth can be designed and used in ultrasonic cleaning, ultrasonic sonochemistry and other applications. PMID- 15759687 TI - Sound-field reproduction in-room using optimal control techniques: simulations in the frequency domain. AB - This paper describes the simulations and results obtained when applying optimal control to progressive sound-field reproduction (mainly for audio applications) over an area using multiple monopole loudspeakers. The model simulates a reproduction system that operates either in free field or in a closed space approaching a typical listening room, and is based on optimal control in the frequency domain. This rather simple approach is chosen for the purpose of physical investigation, especially in terms of sensing microphones and reproduction loudspeakers configurations. Other issues of interest concern the comparison with wave-field synthesis and the control mechanisms. The results suggest that in-room reproduction of sound field using active control can be achieved with a residual normalized squared error significantly lower than open loop wave-field synthesis in the same situation. Active reproduction techniques have the advantage of automatically compensating for the room's natural dynamics. For the considered cases, the simulations show that optimal control results are not sensitive (in terms of reproduction error) to wall absorption in the reproduction room. A special surrounding configuration of sensors is introduced for a sensor-free listening area in free field. PMID- 15759688 TI - Mathematical model for characterizing noise transmission into finite cylindrical structures. AB - This work presents a theoretical study of the sound transmission into a finite cylinder under coupled structural and acoustic vibration. Particular attention of this study is focused on evaluating a dimensionless quantity, "noise reduction," for characterizing noise transmission into a small cylindrical enclosure. An analytical expression of the exterior sound pressure resulting from an oblique plane wave impinging upon the cylindrical shell is first presented, which is approximated from the exterior sound pressure for an infinite cylindrical structure. Next, the analytical solution of the interior sound pressure is computed using modal-interaction theory for the coupled structural acoustic system. These results are then used to derive the analytical formula for the noise reduction. Finally, the model is used to predict and characterize the sound transmission into a ChamberCore cylindrical structure, and the results are compared with experimental data. The effects of incidence angle and internal acoustic damping on the sound transmission into the cylinder are also parametrically studied. PMID- 15759689 TI - Nonlinear vibrations of buried landmines. AB - The seismo-acoustic method is one of the most promising emerging techniques for the detection of landmines. Numerous field tests have demonstrated that buried landmines manifest themselves at the surface through linear and nonlinear responses to acoustic/seismic excitation. The present paper describes modeling of the nonlinear response in the framework of the mass-spring model of the soil-mine system. The perturbation method used in the model allows for the derivation of an analytical solution describing both quadratic and cubic acoustic interactions at the soil-mine interface. This solution has been compared with actual field measurements to obtain nonlinear parameters of the buried mines. These parameters have been analyzed with respect to mine types and burial depths. It was found that the cubic nonlinearity could be a significant contributor to the nonlinear response. This effect has led to the development of a new intermodulation detection algorithm based on dual-frequency excitation. Both quadratic and intermodulation nonlinear algorithms were evaluated at the U.S. Army outdoor testing facilities. The algorithms appear to complement each other in improving the overall detection performance. PMID- 15759690 TI - On the choice of expansion functions in the Helmholtz equation least-squares method. AB - This paper examines the performance of Helmholtz equation least-squares (HELS) method in reconstructing acoustic radiation from an arbitrary source by using three different expansions, namely, localized spherical waves (LSW), distributed spherical waves (DSW), and distributed point sources (DPS), under the same set of measurements. The reconstructed acoustic pressures are validated against the benchmark data measured at the same locations as reconstruction points for frequencies up to 3275 Hz. Reconstruction is obtained by using Tikhonov regularization or its modification with the regularization parameter selected by error-free parameter-choice methods. The impact of the number of measurement points on the resultant reconstruction accuracy under different expansion functions is investigated. Results demonstrate that DSW leads to a better conditioned transfer matrix, yields more accurate reconstruction than both LSW and DPS, and is not affected as much by the change in measurement points. Also, it is possible to obtain optimal locations of the auxiliary sources for DSW, LSW, and DPS by taking an independent layer of measurements. Use of these auxiliary sources and an optimal combination of regularization and error-free parameter choice methods can yield a satisfactory reconstruction of acoustic quantities on the source surfaces as well as in the field in the most cost-effective manner. PMID- 15759691 TI - Krylov subspace iterative methods for boundary element method based near-field acoustic holography. AB - The reconstruction of the acoustic field for general surfaces is obtained from the solution of a matrix system that results from a boundary integral equation discretized using boundary element methods. The solution to the resultant matrix system is obtained using iterative regularization methods that counteract the effect of noise on the measurements. These methods will not require the calculation of the singular value decomposition, which can be expensive when the matrix system is considerably large. Krylov subspace methods are iterative methods that have the phenomena known as "semi-convergence," i.e., the optimal regularization solution is obtained after a few iterations. If the iteration is not stopped, the method converges to a solution that generally is totally corrupted by errors on the measurements. For these methods the number of iterations play the role of the regularization parameter. We will focus our attention to the study of the regularizing properties from the Krylov subspace methods like conjugate gradients, least squares QR and the recently proposed Hybrid method. A discussion and comparison of the available stopping rules will be included. A vibrating plate is considered as an example to validate our results. PMID- 15759692 TI - Helmholtz resonator lined with absorbing material. AB - A closed-form, two-dimensional analytical solution is developed to investigate the acoustic performance of a concentric circular Helmholtz resonator lined with fibrous material. The effect of density and the thickness of the fibrous material in the cavity is examined on the resonance frequency and the transmission loss. With the expressions for the eigenvalue and eigenfunction in the cavity, the transmission loss is obtained for a piston-driven model by applying a pressure/velocity matching technique. The results from the analytical methods are compared to the numerical predictions from a three-dimensional boundary element method and the experimental data obtained from an impedance tube setup. It is shown that the acoustic performance of a Helmholtz resonator may be modified considerably by the density and thickness of the fibrous material without changing the cavity dimensions. PMID- 15759693 TI - A study of active tonal noise control for a small axial flow fan. AB - Sound radiated by a computer cooling fan consists of tones which are phase locked with the rotation, and other less deterministic tones and broadband random noise. This paper demonstrates the feasibility of globally eliminating the rotation locked tones by applying a very simple destructive interference to a modified cooling fan with the number of struts equal to the number of rotor blades. The rig consists of a miniature electret microphone used as a rotation sensor, an ordinary loudspeaker, and a bandpass filter with adjustable amplitude and phase delay. The microphone is located at the inlet bellmouth of the fan to pick up the fluctuating aerodynamic pressure caused by the passing rotor blades. The pressure spectrum is rich in the blade passing frequency (BPF) and its low-order harmonics. It provides much better performance than a pulse-generating tachometer. Analysis of the original fan noise shows that about 90% of the radiated tonal sound is phase locked with rotation, and this portion is almost completely eliminated in all directions. The reductions of the radiated sound power in the first two BPFs are 18.5 and 13.0 dB, respectively, and the overall sound power reduction is 11.0 dB. PMID- 15759694 TI - Inverse method predicting spinning modes radiated by a ducted fan from free-field measurements. AB - In the study the inverse problem of deducing the modal structure of the acoustic field generated by a ducted turbofan is addressed using conventional farfield directivity measurements. The final objective is to make input data available for predicting noise radiation in other configurations that would not have been tested. The present paper is devoted to the analytical part of that study. The proposed method is based on the equations governing ducted sound propagation and free-field radiation. It leads to fast computations checked on Rolls-Royce tests made in the framework of previous European projects. Results seem to be reliable although the system of equations to be solved is generally underdetermined (more propagating modes than acoustic measurements). A limited number of modes are thus selected according to any a priori knowledge of the sources. A first guess of the source amplitudes is obtained by adjusting the calculated maximum of radiation of each mode to the measured sound pressure level at the same angle. A least squares fitting gives the final solution. A simple correction can be made to take account of the mean flow velocity inside the nacelle which shifts the directivity patterns. It consists of modifying the actual frequency to keep the cut-off ratios unchanged. PMID- 15759695 TI - Effective impedance spectra for predicting rough sea effects on atmospheric impulsive sounds. AB - Two methods of calculating the effective impedance spectra of acoustically hard, randomly rough, two-dimensional surfaces valid for acoustic wavelengths large compared with the roughness scales have been explored. The first method uses the complex excess attenuation spectrum due to a point source above a rough boundary predicted by a boundary element method (BEM) and solves for effective impedance roots identified by a winding number integral method. The second method is based on an analytical theory in which the contributions from random distributions of surface scatterers are summed to obtain the total scattered field. Effective impedance spectra deduced from measurements of the complex excess attenuation above 2D randomly rough surfaces formed by semicylinders and wedges have been compared to predictions from the two approaches. Although the analytical theory gives relatively poor predictions, BEM-deduced effective impedance spectra agree tolerably well with measured data. Simple polynomials have been found to fit BEM deduced spectra for surfaces formed by intersecting parabolas corresponding to average roughness heights between 0.25 and 7.5 m and for five incidence angles for each average height. Predicted effects of sea-surface roughness on sonic boom profiles and rise time are comparable to those due to turbulence and molecular relaxation effects. PMID- 15759696 TI - The effect of a periodic absorptive strip arrangement on an interior sound field in a room. AB - In this paper we study the effect of periodically arranged sound absorptive strips on the mean acoustic potential energy density distribution of a room. The strips are assumed to be attached on the room's surface of interest. In order to determine their effect, the mean acoustic potential energy density variation is evaluated as the function of a ratio of the strip's arrangement period to wavelength. The evaluation demonstrates that the mean acoustic potential energy density tends to converge. In addition, a comparison with a case in which absorptive materials completely cover the selected absorptive plane shows that a periodic arrangement that uses only half of the absorptive material can be more efficient than a total covering, unless the frequency of interest does not coincide with the room's resonant frequencies. Consequently, the results prove that the ratio of the arrangement period to the wavelength plays an important role in the effectiveness of a periodic absorptive strip arrangement to minimize a room's mean acoustic potential energy density. PMID- 15759697 TI - A scale value for the balance inside a historical opera house. AB - In the framework of opera house acoustics, the term "balance" refers to the acoustical competition between the singer on the stage and the orchestra in the pit. The mechanism allowing the operatic singers to be heard over the orchestra has to do with their skill in enhancing the vocal emission by a peculiar use of the formant frequencies. This vital factor is sensed by the listeners and, apart from the obvious sound power ratio of the stage and the pit sources, is the main cue that helps to formulate a subjective impression of the balance. To achieve its objective qualification, two calibrated sound sources can be placed on the stage and in the pit, respectively, and their sound level difference is measured at the listeners' seats. The scope of this work is to investigate the relationship between the subjective impression and the objective indicator of the balance and to develop a scale value for the parameter in the case of a historical opera house. For this scope a set of acoustical data from the Teatro Comunale in Ferrara will be used to create synthetic sound fields with controlled conditions of the balance between the stage and the pit. This methodology employs an anechoic piece for soprano (with piano accompaniment) and is implemented in a dead room equipped with an acoustical rendering system. The sound fields are used to investigate the appropriate balance values by means of listening tests. The results of the scaling exercise show that a suitable range of values can be extracted and that the sound from the stage and the pit is perceived as balanced when the loudness difference between the two is comprised within -2.0 dBA and +2.3 dBA. PMID- 15759698 TI - Intersymbol interference in underwater acoustic communications using time reversal signal processing. AB - Coherent underwater communication is hampered by the time spread inherent to acoustic propagation in the ocean. Because time-reversal signal processing produces pulse compression, communications has been suggested as a natural application of the technique. Passive versions of time-reversal processing use a receive-only array to do combined temporal and spatial matched filtering. It can be shown, however, that the pulse compression it achieves is not perfect and that an equalizer that relies solely on time-reversal processing will have an error floor caused by uncompensated intersymbol interference (ISI). In the present paper, a physics-based model is developed for the uncompensated ISI in a passive time-reversal equalizer. The model makes use of a normal-mode expansion for the acoustic field. The matched-filtering integral is approximated and the intermediate result interpreted using the waveguide invariant. After combining across the array and sampling, formal statistical averages of the soft demodulation output are calculated. The results show how performance scales with bandwidth, with the number and position of array elements, and with the length of the finite impulse response matched filters. Good agreement is obtained between the predicted scaling and that observed in field experiments. PMID- 15759699 TI - Characterization of subwavelength elastic cylinders with the decomposition of the time-reversal operator: theory and experiment. AB - The decomposition of the time-reversal operator provides information on the scattering medium. It has been shown [Chambers and Gautesen, J. Acoust. Soc. Am. 109, 2616-2624 (2001)] that a small spherical scatterer is in general associated with four eigenvalues and eigenvectors of the time-reversal operator. In this paper, the 2D problem of scattering by an elastic cylinder, imbedded in water, measured by a linear array of transducers is considered. In this case, the array response matrix has three nonzero singular values. Experimental results are obtained with linear arrays of transducers and for wires of different diameters smaller that the wavelength. It is shown how the singular value distribution and the singular vectors depend on the elastic velocities cL, cT, the density rho of each wire, and on the density rho0 and velocity c0 of the surrounding fluid. These results offer a new perspective towards solution of the inverse problem by determining more than scattering contrast using conventional array processing like that used in medical ultrasonic imaging. PMID- 15759700 TI - Gibbs sampling for time-delay-and amplitude estimation in underwater acoustics. AB - Multipath arrivals at a receiving sensor are frequently encountered in many signal-processing areas, including sonar, radar, and communication problems. In underwater acoustics, numerous approaches to source localization, geoacoustic inversion, and tomography rely on accurate multipath arrival extraction. A novel method for estimation of time delays and amplitudes of arrivals with maximum a posteriori (MAP) estimation is presented here. MAP estimation is optimal if appropriate statistical models are selected for the data; implementation, requiring maximization of a multidimensional function, is computationally demanding. Gibbs sampling is proposed as an efficient means for estimating necessary posterior probability distributions, bypassing analytical calculations. The Gibbs sampler includes as unknowns time delays, amplitudes, noise variance, and number of arrivals. Through Monte Carlo simulations, the method is shown to have a performance very close to that of analytical MAP estimation. The method is also shown to be superior to expectation-maximization, which is often applied to time-delay estimation. The Gibbs sampling approach is demonstrated to be more informative than other time-delay estimation methods, providing complete posterior distributions compared to just point estimates; the distributions capture the uncertainty in the problem, presenting likely values of the unknowns that are different from simple point estimates. PMID- 15759701 TI - Inversion of sonobuoy data from shallow-water sites with simulated annealing. AB - An enhanced simulated annealing algorithm is used to invert sparsely sampled seismic data collected with sonobuoys to obtain seafloor geoacoustic properties at two littoral marine environments as well as for a synthetic data set. Inversion of field data from a 750-m water-depth site using a water-gun sound source found a good solution which included a pronounced subbottom reflector after 6483 iterations over seven variables. Field data from a 250-m water-depth site using an air-gun source required 35,421 iterations for a good inversion solution because 30 variables had to be solved for, including the shot-to receiver offsets. The sonobuoy derived compressional wave velocity-depth (Vp-Z) models compare favorably with Vp-Z models derived from nearby, high-quality, multichannel seismic data. There are, however, substantial differences between seafloor reflection coefficients calculated from field models and seafloor reflection coefficients based on commonly used Vp regression curves (gradients). Reflection loss is higher at one field site and lower at the other than predicted from commonly used Vp gradients for terrigenous sediments. In addition, there are strong effects on reflection loss due to the subseafloor interfaces that are also not predicted by Vp gradients. PMID- 15759702 TI - Simultaneous recording of stimulus-frequency and distortion-product otoacoustic emission input-output functions in human ears. AB - Stimulus frequency otoacoustic emission (SFOAE) input-output (I/O) functions were elicited in normal-hearing adults using unequal-frequency primaries in equal level and fixed-suppressor level (Ls) conditions. Responses were repeatable and similar across a range of primary frequency ratios in the fixed-Ls condition. In comparison to equal-frequency primary conditions [Schairer, Fitzpatrick, and Keefe, J. Acoust. Soc. Am. 114, 944-966 (2003)], the unequal-frequency, fixed-Ls condition appears to be more useful for characterizing SFOAE response growth and relating it to basilar-membrane response growth, and for testing the ability to predict audiometric thresholds. Simultaneously recorded distortion-product OAE (DPOAE) I/O functions had higher thresholds than SFOAE I/O functions, and they identified the onset of the nonlinear-distortion mechanism in SFOAEs. DPOAE threshold often corresponded to nonmonotonicities in SFOAE I/O functions. This suggests that the level-dependent nonmonotonicities and associated phase shifts in SFOAE I/O functions were due to varying degrees of cancellation of two sources of SFOAE, such as coherent reflection and distortion mechanisms. Level-dependent noise was observed on-band (at the frequencies of the stimuli) but not off-band, or in the DPOAEs. The variability was observed in ears with normal hearing and ears with cochlear implants. In general, these results indicate the source of the variability is biological, possibly from within the middle ear. PMID- 15759703 TI - On the binding of successive sounds: perceiving shifts in nonperceived pitches. AB - It is difficult to hear out individually the components of a "chord" of equal amplitude pure tones with synchronous onsets and offsets. In the present study, this was confirmed using 300-ms random (inharmonic) chords with components at least 1/2 octave apart. Following each chord, after a variable silent delay, listeners were presented with a single pure tone which was either identical to one component of the chord or halfway in frequency between two components. These two types of sequence could not be reliably discriminated from each other. However, it was also found that if the single tone following the chord was instead slightly (e.g., 1/12 octave) lower or higher in frequency than one of its components, the same listeners were sensitive to this relation. They could perceive a pitch shift in the corresponding direction. Thus, it is possible to perceive a shift in a nonperceived frequency/pitch. This paradoxical phenomenon provides psychophysical evidence for the existence of automatic "frequency-shift detectors" in the human auditory system. The data reported here suggest that such detectors operate at an early stage of auditory scene analysis but can be activated by a pair of sounds separated by a few seconds. PMID- 15759704 TI - The role of visual speech cues in reducing energetic and informational masking. AB - Two experiments compared the effect of supplying visual speech information (e.g., lipreading cues) on the ability to hear one female talker's voice in the presence of steady-state noise or a masking complex consisting of two other female voices. In the first experiment intelligibility of sentences was measured in the presence of the two types of maskers with and without perceived spatial separation of target and masker. The second study tested detection of sentences in the same experimental conditions. Results showed that visual cues provided more benefit for both recognition and detection of speech when the masker consisted of other voices (versus steady-state noise). Moreover, visual cues provided greater benefit when the target speech and masker were spatially coincident versus when they appeared to arise from different spatial locations. The data obtained here are consistent with the hypothesis that lipreading cues help to segregate a target voice from competing voices, in addition to the established benefit of supplementing masked phonetic information. PMID- 15759705 TI - Voice responses to changes in pitch of voice or tone auditory feedback. AB - The present study was undertaken to examine if a subject's voice F0 responded not only to perturbations in pitch of voice feedback but also to changes in pitch of a side tone presented congruent with voice feedback. Small magnitude brief duration perturbations in pitch of voice or tone auditory feedback were randomly introduced during sustained vowel phonations. Results demonstrated a higher rate and larger magnitude of voice F0 responses to changes in pitch of the voice compared with a triangular-shaped tone (experiment 1) or a pure tone (experiment 2). However, response latencies did not differ across voice or tone conditions. Data suggest that subjects responded to the change in F0 rather than harmonic frequencies of auditory feedback because voice F0 response prevalence, magnitude, or latency did not statistically differ across triangular-shaped tone or pure tone feedback. Results indicate the audio-vocal system is sensitive to the change in pitch of a variety of sounds, which may represent a flexible system capable of adapting to changes in the subject's voice. However, lower prevalence and smaller responses to tone pitch-shifted signals suggest that the audio-vocal system may resist changes to the pitch of other environmental sounds when voice feedback is present. PMID- 15759707 TI - Effects on the glottal voice source of vocal loudness variation in untrained female and male voices. AB - Subglottal pressure is one of the main voice control factors, controlling vocal loudness. In this investigation the effects of subglottal pressure variation on the voice source in untrained female and male voices phonating at a low, a middle, and a high fundamental frequency are analyzed. The subjects produced a series of /pae/ syllables at varied degrees of vocal loudness, attempting to keep pitch constant. Subglottal pressure was estimated from the oral pressure during the /p/ occlusion. Ten subglottal pressure values, approximately equidistantly spaced within the pressure range used, were identified, and the voice source of the vowels following these pressure values was analyzed by inverse filtering the airflow signal as captured by a Rothenberg mask. The maximum flow declination rate (MFDR) was found to increase linearly with subglottal pressure, but a given subglottal pressure produced lower values for female than for male voices. The closed quotient increased quickly with subglottal pressure at low pressures and slowly at high pressures, such that the relationship can be approximated by a power function. For a given subglottal pressure value, female voices reached lower values of closed quotient than male voices. PMID- 15759706 TI - Lip kinematics in long and short stop and fricative consonants. AB - This paper examines lip and jaw kinematics in the production of labial stop and fricative consonants where the duration of the oral closure/constriction is varied for linguistic purposes. The subjects were speakers of Japanese and Swedish, two languages that have a contrast between short and long consonants. Lip and jaw movements were recorded using a magnetometer system. Based on earlier work showing that the lips are moving at a high velocity at the oral closure, it was hypothesized that speakers could control closure/constriction duration by varying the position of a virtual target for the lips. According to this hypothesis, the peak vertical position of the lower lip during the oral closure/constriction should be higher for the long than for the short consonants. This would result in the lips staying in contact for a longer period. The results show that this is the case for the Japanese subjects and one Swedish subject who produced non-overlapping distributions of closure/ constriction duration for the two categories. However, the peak velocity of the lower lip raising movement did not differ between the two categories. Thus if the lip movements in speech are controlled by specifying a virtual target, that control must involve variations in both the position and the timing of the target. PMID- 15759708 TI - Synthesis fidelity and time-varying spectral change in vowels. AB - Recent studies have shown that synthesized versions of American English vowels are less accurately identified when the natural time-varying spectral changes are eliminated by holding the formant frequencies constant over the duration of the vowel. A limitation of these experiments has been that vowels produced by formant synthesis are generally less accurately identified than the natural vowels after which they are modeled. To overcome this limitation, a high-quality speech analysis-synthesis system (STRAIGHT) was used to synthesize versions of 12 American English vowels spoken by adults and children. Vowels synthesized with STRAIGHT were identified as accurately as the natural versions, in contrast with previous results from our laboratory showing identification rates 9%-12% lower for the same vowels synthesized using the cascade formant model. Consistent with earlier studies, identification accuracy was not reduced when the fundamental frequency was held constant across the vowel. However, elimination of time varying changes in the spectral envelope using STRAIGHT led to a greater reduction in accuracy (23%) than was previously found with cascade formant synthesis (11%). A statistical pattern recognition model, applied to acoustic measurements of the natural and synthesized vowels, predicted both the higher identification accuracy for vowels synthesized using STRAIGHT compared to formant synthesis, and the greater effects of holding the formant frequencies constant over time with STRAIGHT synthesis. Taken together, the experiment and modeling results suggest that formant estimation errors and incorrect rendering of spectral and temporal cues by cascade formant synthesis contribute to lower identification accuracy and underestimation of the role of time-varying spectral change in vowels. PMID- 15759709 TI - Lexical frequency and neighborhood density effects on the recognition of native and Spanish-accented words by native English and Spanish listeners. AB - This study examined the effect of presumed mismatches between speech input and the phonological representations of English words by native speakers of English (NE) and Spanish (NS). The English test words, which were produced by a NE speaker and a NS speaker, varied orthogonally in lexical frequency and neighborhood density and were presented to NE listeners and to NS listeners who differed in English pronunciation proficiency. It was hypothesized that mismatches between phonological representations and speech input would impair word recognition, especially for items from dense lexical neighborhoods which are phonologically similar to many other words and require finer sound discrimination. Further, it was assumed that L2 phonological representations would change with L2 proficiency. The results showed the expected mismatch effect only for words from dense neighborhoods. For Spanish-accented stimuli, the NS groups recognized more words from dense neighborhoods than the NE group did. For native-produced stimuli, the low-proficiency NS group recognized fewer words than the other two groups. The-high proficiency NS participants' performance was as good as the NE group's for words from sparse neighborhoods, but not for words from dense neighborhoods. These results are discussed in relation to the development of phonological representations of L2 words. (200 words). PMID- 15759711 TI - Modal analysis and intensity of acoustic radiation of the kettledrum. AB - The acoustical features of kettledrums have been analyzed by means of modal analysis and acoustic radiation (p/v ratio) measurements. Modal analysis of two different kettledrums was undertaken, exciting the system both by a hammer and a shaker. Up to 15 vibrational modes were clearly identified. Acoustic radiation was studied using two ways. Based on previous experiments of other researchers, a new parameter, called intensity of acoustic radiation (IAR), has been defined and measured. Results show a strict relationship between IAR and the frequency response function (FRF, which is the v/F ratio), and IAR also strongly relates the modal pattern to acoustic radiation. Finally, IAR is proposed for vibro acoustical characterization of kettledrums and other musical instruments such as strings, pianos, and harpsichords. PMID- 15759710 TI - A statistics-based pitch contour model for Mandarin speech. AB - A statistics-based syllable pitch contour model for Mandarin speech is proposed. This approach takes the mean and the shape of a syllable log-pitch contour as two basic modeling units and considers several affecting factors that contribute to their variations. The affecting factors include the speaker, prosodic state (which essentially represents the high-level linguistic components of F0 and will be explained more clearly in Sec. I), tone, and initial and final syllable classes. The parameters of the two modeling units were automatically estimated using the expectation-maximization (EM) algorithm. Experimental results showed that the root mean squared errors (RMSEs) obtained in the closed and open tests in the reconstructed pitch period were 0.362 and 0.373 ms, respectively. This model provides a way to separate the effects of several major factors. All of the inferred values of the affecting factors were in close agreement with our prior linguistic knowledge. It also gives a quantitative and more complete description of the coarticulation effect of neighboring tones rather than conventional qualitative descriptions of the tone sandhi rules. In addition, the model can provide useful cues to determine the prosodic phrase boundaries, including those occurring at intersyllable locations, with or without punctuation marks. PMID- 15759712 TI - High-frequency ultrasound scattering from microspheres and single cells. AB - Assessing the proportion of biological cells in a volume of interest undergoing structural changes, such as cell death, using high-frequency ultrasound (20-100 MHz), requires the development of a theoretical model of scattering by any arbitrary cell ensemble. A prerequisite to building such a model is to know the scattering by a single cell in different states. In this paper, a simple model for the high-frequency acoustic scattering by one cell is proposed. A method for deducing the backscatter transfer function from a single, subresolution scatterer is also devised. Using this method, experimental measurements of backscatter from homogeneous, subresolution polystyrene microspheres and single, viable eukaryotic cells, acquired across a broad, continuous range of frequencies were compared with elastic scattering theory and the proposed cell scattering model, respectively. The resonant features observed in the backscatter transfer function of microspheres were found to correspond accurately to theoretical predictions. Using the spacing of the major spectral peaks in the transfer functions obtained experimentally, it is possible to predict microsphere diameters with less than 4% error. Such good agreement was not seen between the cell model and the measured backscatter from cells. Possible reasons for this discrepancy are discussed. PMID- 15759713 TI - Pitch (F0) and formant profiles of human vowels and vowel-like baboon grunts: the role of vocalizer body size and voice-acoustic allometry. AB - Key voice features--fundamental frequency (F0) and formant frequencies--can vary extensively between individuals. Much of the variation can be traced to differences in the size of the larynx and vocal-tract cavities, but whether these differences in turn simply reflect differences in speaker body size (i.e., neutral vocal allometry) remains unclear. Quantitative analyses were therefore undertaken to test the relationship between speaker body size and voice F0 and formant frequencies for human vowels. To test the taxonomic generality of the relationships, the same analyses were conducted on the vowel-like grunts of baboons, whose phylogenetic proximity to humans and similar vocal production biology and voice acoustic patterns recommend them for such comparative research. For adults of both species, males were larger than females and had lower mean voice F0 and formant frequencies. However, beyond this, F0 variation did not track body-size variation between the sexes in either species, nor within sexes in humans. In humans, formant variation correlated significantly with speaker height but only in males and not in females. Implications for general vocal allometry are discussed as are implications for speech origins theories, and challenges to them, related to laryngeal position and vocal tract length. PMID- 15759714 TI - Automatic classification and speaker identification of African elephant (Loxodonta africana) vocalizations. AB - A hidden Markov model (HMM) system is presented for automatically classifying African elephant vocalizations. The development of the system is motivated by successful models from human speech analysis and recognition. Classification features include frequency-shifted Mel-frequency cepstral coefficients (MFCCs) and log energy, spectrally motivated features which are commonly used in human speech processing. Experiments, including vocalization type classification and speaker identification, are performed on vocalizations collected from captive elephants in a naturalistic environment. The system classified vocalizations with accuracies of 94.3% and 82.5% for type classification and speaker identification classification experiments, respectively. Classification accuracy, statistical significance tests on the model parameters, and qualitative analysis support the effectiveness and robustness of this approach for vocalization analysis in nonhuman species. PMID- 15759715 TI - Acoustic characterization in whole blood and plasma of site-targeted nanoparticle ultrasound contrast agent for molecular imaging. AB - The ability to enhance specific molecular markers of pathology with ultrasound has been previously demonstrated by our group employing a nanoparticle contrast agent [Lanza et al., Invest. Radiol. 35, 227-234 (2000); Ultrasound Med. Biol. 23, 863-870 (1997)]. One of the advantages of this agent is very low echogenicity in the blood pool that allows increased contrast between the blood pool and the bound, site-targeted agent. We measured acoustic backscatter and attenuation coefficient as a function of the contrast agent concentration, ambient pressure, peak acoustic pressure, and as an effect of duty cycle and wave form shape. Measurements were performed while the nanoparticles were suspended in either whole porcine blood or plasma. The nanoparticles were only detectable when insonified within plasma devoid of red blood cells and were shown to exhibit backscatter levels more than 30 dB below the backscatter from whole blood. Attenuation of nanoparticles in whole porcine blood was not measurably different from that of whole blood alone over a range of concentrations up to eight times the maximum in vivo dose. The resulting data provide upper bounds on blood pool attenuation coefficient and backscatter and will be needed to more precisely define levels of molecular contrast enhancement that may be obtained in vivo. PMID- 15759716 TI - Quilliam wins 2005 Wiley Award. PMID- 15759717 TI - Evaluation study protocols and acceptance criteria for hand held assays intended for detection of Bacillus anthracis. PMID- 15759718 TI - A field guide to instrumentation. PMID- 15759719 TI - Identification and estimation of the levo isomer in raw materials and finished products containing atropine and/or hyoscyamine. AB - The belladonna alkaloids atropine sulfate and hyoscyamine sulfate, occasionally used as anticholinergic and antimuscarinic agents, have identical molecular formulas but different stereo configurations. Hyoscyamine sulfate contains almost 100% of the levo isomer, whereas atropine sulfate is composed of equal parts of dextro and levo isomers. It is believed that the therapeutic properties of these alkaloids are due exclusively or primarily to the levo isomer. Currently available methods determine only the total amount of atropine (hyoscyamine) sulfate. A method has been developed and is reported for the identification and estimation of the levo and dextro isomers of atropine and hyoscyamine. Reference solutions are prepared in methanol at the following weights per 100 mL: 8.0 mg atropine sulfate; 4.0 mg hyoscyamine sulfate; 7.0 mg scopolamine hydrobromide; and 10.0 mg homatropine methylbromide. Samples of raw materials are similarly prepared in methanol, commercial products are also extracted or diluted with methanol, and solutions are filtered. Liquid chromatography is used for separations on a 25 cm Chirobiotic T2 column. The mobile phase is prepared by mixing 3.0 mL acetic acid and 2.0 mL triethylamine with 1000 mL methanol. The injection volume is 100 or 200 microL; the flow rate is about 0.35 mL/min. Fluorescence detection is at 255 nm excitation and 285 nm emission. Scopolamine hydrobromide and hyoscyamine eluted after 20 and 60 min, respectively. Atropine sulfate generated 2 peaks after 60 and 65 min. Homatropine methylbromide also produced 2 peaks after 70 and 85 min. Samples tested in this study included raw materials and commercial tablets or injections containing belladonna alkaloids. In all cases, the percentage calculated was that of the levo isomer relative to the total amount of atropine (hyoscyamine) present. PMID- 15759720 TI - Microbiological assay-trienzyme procedure for total folates in cereals and cereal foods: collaborative study. AB - In 1996, U.S. Food and Drug Administration regulations mandated the fortification of enriched cereal-grain products with folic acid, thereby emphasizing the need for validated methods for total folates in foods, particularly cereal products. The AOAC Official Methods (944.12, 960.46) currently used for the analysis of folate in foods for compliance purposes are microbiological methods. When the fortification regulations were finalized, no Official AOAC or Approved AACC methods for folate in cereal-grain products were in place. The AOAC Official Method (992.05) for folic acid in infant formula does not incorporate important improvements in the extraction procedure and was not considered suitable for the analysis of folates in foods in general. A microbiological assay protocol using a trienzyme extraction procedure was prepared and submitted for comments to 40 laboratories with recognized experience in folate analysis. On the basis of comments, the method was revised to have the conjugase (gamma-glutamyl-carboxy peptidase) treatment follow a protease treatment, to include the use of cryoprotected inoculum, and to include the spectroscopic standardization of the standard and optional use of microtiter plates. Thirteen laboratories participated in a collaborative study of 10 required and 10 optional cereal-grain products, including flour, bread, cookies, baking mixes, and ready-to-eat breakfast cereals. The majority of the participating laboratories performed the assay by the standard test tube method; others used the microtiter plate modification for endpoint quantitation with equal success. For the required products, the relative standard deviation between laboratories (RSD(R)) ranged from 7.4 to 21.6% for 8 fortified (or enriched) products compared with expected (Horwitz equation-based) values of 11-20%. RSD(R) values were higher (22.7-52.9%) for 2 unfortified cereal-grain products. For the optional products, the RSD(R) ranged from 1.8 to 11.2% for 8 fortified products. RSD(R) values were higher (27.9-28.7%) for 2 unfortified cereal-grain products. Based on the results of the collaborative study, the microbiological assay with trienzyme extraction is recommended for adoption as Official First Action. PMID- 15759721 TI - Determination of six kavalactones in dietary supplements and selected functional foods containing Piper methysticum by isocratic liquid chromatography with internal standard. AB - Kava (Piper methysticum) dietary products have been sold worldwide for treatment of nervous anxiety, tension, and restlessness. Recent reports showed potential association of kava usage and liver injuries. This study was conducted to develop simple and reliable methodologies for the extraction and determination of 6 major kavalactones: (+)-methysticin, (+)-dihydromethysticin, (+)-kavain, (+) dihydrokavain, yangonin, and desmethoxyyangonin. Ultrasonic extraction techniques and isocratic reversed-phase liquid chromatography (LC) were optimized for different types of samples, including capsules containing kava root extract or root powder, raw root material, tea bags, and snack bar. A suitable internal standard, 5,7-dihydroxyflavone, was used for LC calibration. Kavalactones were completely separated in 30 min using a Luna C18-2 column at 60 degrees C with an isocratic mobile phase consisting of 2-propanol-acetonitrile-water-acetic acid (16 + 16 + 68 + 0.1, v/v/v/v). Within-laboratory, intraday, and interday method variation (% relative standard deviation) for most samples extracted by methanol or methanol-water mixture were <5%. Lower levels of kavalactone contents and higher variations were observed for tea bags from water extraction or infusion as compared to methanol extraction. Labeling information of tea bags based on methanol extraction could be misleading to consumers. Analytical recoveries of snack bar fortified at 10 and 20 microg/g were >84% with RSD values <8%. Methods developed in this study offer a simple and reproducible means for analysis of kavalactones in various matrixes of dietary products. PMID- 15759722 TI - Improved extraction of ginkgotoxin (4'-O-methylpyridoxine) from Ginkgo biloba products. AB - A simple extraction procedure was applied to the analysis of canned/packaged white nuts and Ginkgo biloba extracts. Extraction by shaking with water at room temperature was more convenient to use than a previously published Soxhlet procedure for analysis of packaged Ginkgo biloba seeds (white nuts) for ginkgotoxin; recoveries from spiked dried seeds by the simple extraction procedure averaged 76%. Determination was by liquid chromatography with UV or fluorescence detection. Recoveries of ginkgotoxin from a spiked and unspiked natural health product (powder from Ginkgo biloba capsules) were equivalent by both procedures; recovery from spiked powder by the simple extraction procedure was 81%. Application of this extraction procedure in the analysis of 6 samples of white nuts (vacuum packaged and canned products) showed that free ginkgotoxin was present in 5 samples at concentrations up to 25 microg/g dry weight. Total ginkgotoxin was determined after hydrolysis with beta-glucosidase of sample extracts in which a peak corresponding to the 5'-O-glucoside was detected. Ginkgotoxin was determined in 10 Ginkgo biloba natural health products by the same method at levels up to 181 microg/g. PMID- 15759723 TI - Liquid chromatographic analysis of vitamin B6 in reconstituted infant formula: collaborative study. AB - A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in infant formula. Total vitamin B6 was quantified by converting the phosphorylated and free vitamers into pyridoxine. Pyridoxine was determined by ion pair reversed-phase LC with fluorescence detection. The method was subjected to an AOAC collaborative study involving a factory-manufactured, milk- and soy based infant formula. Each was spiked at 3 concentrations in the range of 0-1 microg/g and sent as blind duplicate to participant laboratories. Nine laboratories returned valid data which were statistically analyzed for outliers and precision parameters. The repeatability relative standard deviation (RSD(r)) ranges were 2.0-4.0 and 3.5-5.9% for fortified milk- and soy-based formulas, respectively. The reproducibility relative standard deviation (RSD(R)) ranges were 8.2-8.4 and 6.7-11.2% for fortified milk- and soy-based formulas, respectively. HORRAT values ranged from 0.42 to 0.53, indicating that the precision of the method is acceptable. The mean RSD(r):RSD(R) values were 0.60 and 0.55 for milk- and soy-based formulas, respectively. As expected, RSDs for the unfortified samples were higher, but their HORRAT values (0.81 and 2.06) helped define a realistic limit of quantitation as 0.05 microg/g. Recovery data were quantitative and varied between 81.4 and 98.0% (mean = 89.8%) for each of 6 spiked materials. PMID- 15759724 TI - Development and validation of spectrophotometric methods for determination of fluoxetine, sertraline, and paroxetine in pharmaceutical dosage forms. AB - Three simple and sensitive spectrophotometric methods were developed and validated for determination of the hydrochloride salts of fluoxetine, sertraline, and paroxetine in their pharmaceutical dosage forms. These methods were based on the reaction of the N-alkylvinylamine formed from the interaction of the free secondary amino group in the investigated drugs and acetaldehyde with each of 3 haloquinones, i.e., chloranil, bromanil, and 2,3-dichloronaphthoquinone, to give colored vinylamino-substituted quinones. The colored products obtained with chloranil, bromanil, and 2,3-dichloronaphthoquinone exhibit absorption maxima at 665, 655, and 580 nm, respectively. The factors affecting the reactions were studied and optimized. Under the optimum reaction conditions, linear relationships with good correlation coefficients (0.9986-0.9999) were found between the absorbances and the concentrations of the investigated drugs in the range of 4-120 microg/mL. The limits of detection for the assays ranged from 1.19 to 2.98 microg/mL. The precision values of the methods were satisfactory; the relative standard deviations were 0.56-1.24%. The proposed methods were successfully applied to the determination of the 3 drugs in pure and pharmaceutical dosage forms with good accuracy; the recoveries ranged from 99.1 to 101.3% with standard deviations of 1.15-1.92%. The results compared favorably with those of reported methods. PMID- 15759725 TI - Determination of beta-agonist residues in bovine urine using liquid chromatography-tandem mass spectrometry. AB - A multiresidue method was developed and validated to screen bovine urine samples for 10 beta-2-adrenergic agonistic drugs--brombuterol, cimaterol, clenbuterol, clenpenterol, isoxsuprine, mabuterol, ractopamine, ritodrine, salbutamol, and tulobuterol--at the 2 microg/L level. The method is also quantitative in the range of 1 to 4 microg/L for all analytes except salbutamol. The procedure uses enzymatic digestion, liquid-liquid extraction, and cleanup on solid-phase extraction columns, followed by detection using a liquid chromatograph-tandem quadrupole mass spectrometer operated in the positive-ion atmospheric pressure chemical ionization multiple-reaction monitoring mode. Method validation included assessment of recoveries, repeatabilities, linearity of responses, decision limits, and detection capabilities. Overall average recoveries ranged from 70 91%; recoveries were generally lower for salbutamol. The decision limits ranged from 0.4-1.0 microg/L, and detection capabilities from 0.6-1.7 microg/L. PMID- 15759726 TI - Determination of chloramphenicol residue in fish and shrimp tissues by gas chromatography with a microcell electron capture detector. AB - A gas chromatography method with microcell electron capture detection was developed for the determination of chloramphenicol residue in fish and shrimp muscle tissues. The tissue samples were extracted with ethyl acetate, defatted with hexane, and derivatized with Sylon BFT [N,O-bis (trimethylsilyl) trifluoroacetamide-trimethylchlorosilane (99 + 1)]. The limit of detection was 0.04 ng/g and the limit of quantitation 0.1 ng/g. Average recoveries were 70.8 90.8% for fish and 69.9-86.3% for shrimp, respectively. The method was validated for the determination of practical samples. PMID- 15759727 TI - Determination of beta-agonists in liver and retina by liquid chromatography tandem mass spectrometry. AB - A liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the determination of bromobuterol, cimaterol, clenbuterol, clenpenterol, hydroxymethylclenbuterol, isoxsuprine, mabuterol, ractopamine, ritrodrine, salbutamol, terbutaline, and tulobuterol residues in bovine liver and retina is reported. This procedure uses enzymatic digestion, liquid-liquid extraction, and cleanup on Oasis HLB solid-phase extraction cartridges, followed by determination of the residues by LC-tandem quadrupole MS using atmospheric pressure chemical ionization in the positive ion mode. Overall average recoveries ranged from 23 to 76% for liver and 34 to 77% for retina. The mean values for samples fortified at levels between 0.5-2.0 microg/kg (liver) and 5-20 microg/kg (retina) agreed within 98-118% of the spiked levels, with coefficients of variation ranging from 6 to 20%. The decision limits, CCalpha, ranged from 0.1 to 0.3 microg/kg for liver, 1-3 microg/kg for retina, and detection capabilities, CCbeta, from 0.2-0.5 microg/kg for liver and 2-5 microg/kg for retina. PMID- 15759729 TI - Stability-indicating methods for determination of pyritinol dihydrochloride in the presence of its precursor and degradation product by derivative spectrophotometry. AB - A first-derivative spectrophotometric (1D) method and a derivative-ratio zero crossing spectrophotometric (1DD) method were used to determine pyritinol dihydrochloride (I) in the presence of its precursor (II) and its degradation product (III) with 0.1N hydrochloric acid as a solvent. Linear relationships were obtained in the ranges of 6-22 microg/mL for the (1D) method and 6-20 microg/mL for the (1DD) method. By applying the proposed methods, it was possible to determine pyritinol dihydrochloride in its pure powdered form with an accuracy of 100.36 +/- 1.497% (n = 9) for the (1D) method and an accuracy of 99.92 +/- 1.172% (n = 8) for the (1DD) method. Laboratory-prepared mixtures containing different ratios of (I), (II), and (III) were analyzed, and the proposed methods were valid for concentrations of < or = 10% (II) and < or = 50% (III). The proposed methods were validated and found to be suitable as stability-indicating assay methods for pyritinol in pharmaceutical formulations. PMID- 15759728 TI - Densitometric high performance thin-layer chromatography identification and quantitative analysis of psychotropic drugs. AB - A thin-layer chromatography (TLC)-densitometry method has been developed to identify and quantify haloperidol, amitriptyline, sulpiride, promazine, fluphenazine, doxepin, diazepam, trifluoperazine, clonazepam, and chlorpromazine in selected psychotropic drugs. Separation was performed on precoated silica gel 60 F254 TLC plates. Chromatograms were developed in various mobile phases, and 8 of 30 tested phases were selected based on spot location and developing time. The identification and quantification were carried out based on ultraviolet densitometric measurements at chosen wavelengths. In addition to retention coefficients, the absorption spectra recorded directly from chromatograms were also used in qualitative analysis. Under established experimental conditions, high sensitivity of the method was achieved. The limit of detection ranged from 0.009 to 0.260 microg, depending on the wavelength selected for measuring. A satisfactory recovery, ranging from 92.99 to 104.70%, was achieved for individual constituents. PMID- 15759730 TI - Confirmation of synthetic glucocorticoids with liquid chromatography/mass spectrometry: organization and results of an international interlaboratory comparison test. AB - Within the framework of a European Union (EU) research project entitled "Food Safety Screening: Synthetic Glucocorticoids (QLK1-1999-00122)," an international interlaboratory ring test was organized to compare and evaluate different liquid chromatography/mass spectrometry (LC/MS) confirmatory methods that are applied in European monitoring programs for detecting the use of synthetic glucocorticoids. Liver and urine samples of bovines treated with synthetic glucocorticoids were collected and sent to the participants of the study for analysis. Participants received 3 liver and 3 urine samples and were free to use either their own LC/MS method or an LC/MS-based method developed during the EU research project. The residue concentrations in the samples were calculated as the mean of the concentrations reported by each laboratory. The mean dexamethasone concentration of liver sample L1 was calculated as 2.27 microg/kg [relative standard deviation (RSD) 43%, n = 9], which exceeds the maximum residue level (MRL) of 2 microg/kg. Three of the 9 laboratories (33%) reported concentration levels less than 2 microg/kg, resulting in obviously false compliant results. The overall mean concentration of flumethasone in liver sample L2 was calculated as 3.27 microg/kg (RSD 33%, n = 8). Applying a comparable limit for flumethasone of 2 microg/kg, 8 of the 9 laboratories would have obtained a correct noncompliant result. As for the blank liver sample, 1 participant found a false noncompliant result. The urine sample U1 contained prednisolone residues at a mean concentration of 1.58 microg/kg (RSD 43%, n = 9). Four out of 9 results were less than a theoretical minimum required performance level (MRPL) of 2 microg/kg. The calculated concentration of dexamethasone in urine sample U3 was 5.21 microg/kg (RSD 62%, n = 9). One of the 9 results was lower than 2 microg/kg. Urine sample U2 was correctly reported as blank by all participants. PMID- 15759731 TI - Validation of a method for the determination of narasin in the edible tissues of chickens by liquid chromatography. AB - Maxiban and Monteban are 2 products marketed by Elanco Animal Health. They contain narasin and are used for the prevention of coccidiosis in chickens. Products used in the European market must be regularly re-registered with new data to support label claims. This study was undertaken as part of such a re registration effort. A method for the determination of narasin in poultry tissue was previously registered with the authorities; however, a method with more environmentally friendly solvents was desired. The Canadian Food Inspection Agency accomplished this goal and published an improved method. In order to register the method with European authorities as the official Elanco method for determination of narasin, Elanco scientists were required to provide validation data for all edible poultry tissues. This paper shows the validation of the method to detect residues of narasin using solid-phase extraction followed by liquid chromatographic analysis utilizing post-column derivatization. PMID- 15759732 TI - Furan precursors in food: a model study and development of a simple headspace method for determination of furan. AB - Furan was previously detected in foods that had undergone thermal treatment. Because furan is now classified as a possible human carcinogen, a model system was developed to investigate the origins of furan. Also, a simple, rapid isotope dilution (d4-furan) headspace method was developed to measure furan. Two pathways of furan formation have been identified in the model systems tested so far. The first is the oxidation of polyunsaturated fatty acids at elevated temperatures, and the second is linked to the decomposition of ascorbic acid derivatives. The analytical procedure, based on the use of a 50 microL injection (from the headspace of a 1.5 mL vial containing 0.5 mL water) into the split/splitless injection port of a gas chromatograph/mass spectrometer (electron ionization, selected-ion monitoring), showed linearity in the 10-1000 ng/g range with a limit of detection of 1 ng/g. PMID- 15759733 TI - Rapid determination of moisture/solids and fat in dairy products by microwave and nuclear magnetic resonance analysis. AB - A peer-verified method is presented for the determination of percent moisture/solids and fat in dairy products by microwave drying and nuclear magnetic resonance (NMR) analysis. The method involves determining the moisture/solids content of dairy samples by microwave drying and using the dried sample to determine the fat content by NMR analysis. Both the submitting and peer laboratories analyzed various dairy products by using a CEM SMART system (moisture) and the SMART Trac (fat). The samples included milks, creams, ice cream mix, sour cream, yogurt, cream cheese, and mozzarella, Swiss, and cheddar cheeses. These samples represented a range of products that processors deal with in daily plant operations. The results were compared with moisture/solids and fat values derived from AOAC-approved methods. PMID- 15759734 TI - Single laboratory validation of a method for the determination of hydroxymethylfurfural in honey by using solid-phase extraction cleanup and liquid chromatography. AB - A method is described for the determination of hydroxymethylfurfural (HMF) in honey. The method, which is based on solid-phase extraction cleanup followed by liquid chromatography (LC) with UV absorbance detection, was tested on a variety of different honey types: liquid, set, blended, filtered, crystalline, and comb honey. A sample of honey fortified with a known amount of HMF acted as an in house reference material. LC with diode-array detection showed that the HMF peak did not contain any peaks of coeluting interfering species. Stability studies showed that honey samples should not be repeatedly frozen and thawed because the temperature changes caused a gradual increase in the HMF concentration. It was also shown that aqueous HMF standard solutions should be kept in the dark at 4 degrees C to avoid degradation of the HMF. The method was internally validated, and the measurement uncertainty was estimated to be +/-9.0 at 40 mg/kg, the legal limit. A comparison of the relative standard uncertainty with the Horwitz relative standard deviation showed that the method was suitable for its purpose and should be validated by a collaborative trial. PMID- 15759735 TI - Polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial cytb gene from species of dairy interest. AB - Species identification plays an important role in food allergy prevention and food substitution detection that can reduce the commercial value of a product. For these reasons, many molecular methods have been developed to determine species origin; among them, polymerase chain reaction (PCR)-based methods were successfully applied to processed or unprocessed foodstuffs. An updated PCR-RFLP (restriction fragment length polymorphism) method of the cytb gene was developed for the identification of the 4 species of main interest in the dairy industry (Bos, Ovis, Capra, Bubalus). The comparative analysis of the 92 cytb sequences available in the database belonging to the 4 species allowed identification of 2 highly conserved regions, which were used to design 2 oligonucleotides for the PCR amplification of a 275 base-pair (bp) cytb fragment. The in silico analysis allowed identification of a set of species-specific restriction endonucleases (HaeIII, TaqI, and MwoI), which generated easily analyzable species-specific restriction profiles of the 275 bp cytb DNA fragment. The system was developed for both purified DNA and DNA extracted from meat or dairy products and finally tested on mixed samples, indicating its applicability to foodstuffs. PMID- 15759736 TI - Polymerase chain reaction technology as analytical tool in agricultural biotechnology. AB - The agricultural biotechnology industry applies polymerase chain reaction (PCR) technology at numerous points in product development. Commodity and food companies as well as third-party diagnostic testing companies also rely on PCR technology for a number of purposes. The primary use of the technology is to verify the presence or absence of genetically modified (GM) material in a product or to quantify the amount of GM material present in a product. This article describes the fundamental elements of PCR analysis and its application to the testing of grains. The document highlights the many areas to which attention must be paid in order to produce reliable test results. These include sample preparation, method validation, choice of appropriate reference materials, and biological and instrumental sources of error. The article also discusses issues related to the analysis of different matrixes and the effect they may have on the accuracy of the PCR analytical results. PMID- 15759737 TI - Performance Tested Method multiple laboratory validation study of ELISA-based assays for the detection of peanuts in food. AB - Performance Tested Method multiple laboratory validations for the detection of peanut protein in 4 different food matrixes were conducted under the auspices of the AOAC Research Institute. In this blind study, 3 commercially available ELISA test kits were validated: Neogen Veratox for Peanut, R-Biopharm RIDASCREEN FAST Peanut, and Tepnel BioKits for Peanut Assay. The food matrixes used were breakfast cereal, cookies, ice cream, and milk chocolate spiked at 0 and 5 ppm peanut. Analyses of the samples were conducted by laboratories representing industry and international and U.S governmental agencies. All 3 commercial test kits successfully identified spiked and peanut-free samples. The validation study required 60 analyses on test samples at the target level 5 microg peanut/g food and 60 analyses at a peanut-free level, which was designed to ensure that the lower 95% confidence limit for the sensitivity and specificity would not be <90%. The probability that a test sample contains an allergen given a prevalence rate of 5% and a positive test result using a single test kit analysis with 95% sensitivity and 95% specificity, which was demonstrated for these test kits, would be 50%. When 2 test kits are run simultaneously on all samples, the probability becomes 95%. It is therefore recommended that all field samples be analyzed with at least 2 of the validated kits. PMID- 15759738 TI - Immunochemical analytical methods for the determination of peanut proteins in foods. AB - Peanut proteins can cause allergenic reactions that can result in respiratory and circulatory effects in the body sometimes leading to shock and death. The determination of peanut proteins in foods by analytical methods can reduce the risk of serious reactions in the highly sensitized individual by allowing for the detection of these proteins in a food at various stages of the manufacturing process. The method performance of 4 commercially available enzyme-linked immunosorbent assay (ELISA) kits was evaluated for the detection of peanut proteins in milk chocolate, ice cream, cookies, and breakfast cereals: ELISA-TEK Peanut Protein Assay, now known as "Bio-Kit" for peanut proteins, from ELISA Technologies Inc.; Veratox for Peanut Allergens from Neogen Corp.; RIDASCREEN Peanut Kit from R-Biopharm GmbH; and ProLisa from Canadian Food Technology Ltd. The 4 test kits were evaluated for accuracy (recovery) and precision using known concentrations of peanut or peanut proteins in the 4 food matrixes. Two different techniques, incurred and spiked, were used to prepare samples with 4 known concentrations of peanut protein. Defatted peanut flour was added in the incurred samples, and water-soluble peanut proteins were added in the spiked samples. The incurred levels were 0.0, 10, 20, and 100 microg whole peanut per g food; the spiked levels were 0.0, 5, 10, and 20 microg peanut protein per g food. Performance varied by test kit, protein concentration, and food matrix. The Veratox kit had the best accuracy or lowest percent difference between measured and incurred levels of 15.7% when averaged across all incurred levels and food matrixes. Recoveries associated with the Veratox kit varied from 93 to 115% for all food matrixes except cookies. Recoveries for all kits were about 50% for cookies. The analytical precision, as measured by the variance, increased with an increase in protein concentration. However, the coefficient of variation (CV) was stable across the 4 incurred protein levels and was 7.0% when averaged across the 4 food matrixes and analytical kits. The R-Biopharm test kit had the best precision or a CV of 4.2% when averaged across all incurred levels and food matrixes. Because measured protein values varied by test kit and food matrix, a method was developed to normalize or transform measured protein concentrations to an adjusted protein value that was equal to the known protein concentration. The normalization method adjusts measured protein values to equal the true protein value regardless of the type test kit or type food matrix. PMID- 15759739 TI - Validation of methods for confirmatory identification of presumptive isolates of Bacillus anthracis. PMID- 15759740 TI - Identification of Bacillus anthracis from culture using gas chromatographic analysis of fatty acid methyl esters. PMID- 15759742 TI - RAMP anthrax test cartridge. PMID- 15759741 TI - Carrier tests to assess microbicidal activities of chemical disinfectants for use on medical devices and environmental surfaces. AB - For well over a decade, many deficiencies have been identified in current AOAC methods used to assess the microbicidal activities of chemical disinfectants on medical devices and environmental surfaces. This report discusses the development of quantitative carrier tests (QCT) designed to address these concerns. Decontamination of surfaces with dried inocula is invariably more difficult than when microorganisms are in suspension. For medical device as well as environmental decontamination, microbicides are used on contaminated surfaces, thus making it necessary to evaluate their microbicidal action on representative carrier materials contaminated with a dried challenge microorganism(s). Our approach is a 2-tiered QCT. The first tier (QCT-1) uses relatively ideal conditions to assess performance of the microbicide for screening purposes; the test uses smooth glass surfaces and quantities of disinfectant in excess of those likely to be experienced in the field. The second tier of testing (QCT-2) is more stringent because it uses (1) disks of brushed stainless steel as carriers, (2) only 50 microL of the test formulation on each carrier as compared to 1 mL in QCT 1, and (3) an added soil load to simulate the presence of residual body fluids or accumulated surface dirt. This review also discusses the factors that affect disinfection of medical devices and environmental surfaces in the context of the methodology used to evaluate the potency of microbicides. Specific recommendations for discussion are included, and performance criteria are suggested based on a risk-reduction approach for different classes of disinfectants. The focus is on improving the relevance of the test methodology to actual field use of disinfectants for devices and facilities in health care, and potentially in other settings. It is hoped that this review and its recommendations will initiate needed discussion and resolution of the many issues identified. PMID- 15759744 TI - Determination of metal content in valerian root phytopharmaceutical derivatives by atomic spectrometry. AB - Phytopharmaceuticals containing Valerian are used as mild sleep-inducing agents. The elemental composition of 3 different marks of Valeriana officinalis roots commercially available in the Argentinian market, their teas, and a commercial tincture have been studied. The content of Al, Ca, Cd, Co, Cr, Cu, Fe, Li, Mn, Ni, Pb, V, and Zn was determined in phytopharmaceuticals by flame atomic emission/absorption spectrometry, electrothermal atomic absorption spectrometry, and ultrasonic nebulization coupled to inductively coupled plasma-optical emission spectrometry. Prior to analyses of the samples, a digestion procedure was optimized. The analytical results obtained for Fe, Al, Ca, and V in the solid sample study were within the range 100-1000 mg/kg, and for Mn, Zn, and Pb within the range 10-100 mg/kg. Cadmium was found at levels up to 0.0125 mg/kg. PMID- 15759745 TI - Acrylamide: considerations for risk management. AB - The presence of acrylamide in many carbohydrate-rich foods is due to its formation during conventional heating and preparation methods. Although acrylamide is established to be a toxic substance, the implications to public health from the amounts found in food are not clear. A better scientific understanding is required to help determine whether, and to what extent, formal risk management action might be necessary. Since acrylamide in food was highlighted in 2002, numerous investigations and initiatives have been developed, including international collaborations across governments, industry, research organizations, and consumer representations. The newly generated information is being used to help the overall understanding of this issue. In particular, new information on health aspects will be important to update the scientific risk assessment. The basis for decisions on possible risk management measures would then be clearer. If future risk assessment concludes that the amounts of acrylamide in food can pose a health threat, then options for risk management will need to be considered, such as limits, guide levels, codes of practice, guidance information, and advice to the food and catering industries and to consumers. In the meantime, it is possible to benefit from progress already made on how acrylamide is formed in food and on ways to lower the amounts present. Raising awareness to the approaches that can reduce the presence of acrylamide in food should be encouraged. Where feasible, such approaches can be assessed for practical use in production, processing, and preparation of the relevant food products. PMID- 15759743 TI - Analytical methods used in the United Kingdom Wildlife Incident Investigation Scheme for the detection of animal poisoning by pesticides. AB - The United Kingdom Wildlife Incident Investigation Scheme (WIIS) investigates cases of suspected poisoning of wildlife, honey bees, and companion animals by pesticides. Together with field inquiries and veterinary post-mortem examinations, the analytical procedures presented here provide a comprehensive approach to the investigation of these cases. The paper covers selection of animal tissues for analysis and methods suitable for the analysis of honey bees and for various types of bait. Seven multiresidue methods cover around 130 pesticides, and methods are also described for a further 8 compounds. These methods are currently used on samples submitted to the Scheme in England and Wales. PMID- 15759746 TI - Acrylamide: update on selected research activities conducted by the European food and drink industry. AB - This paper reviews the progress made by the European food and drink industry (CIAA) on acrylamide with regard to analytical methods, mechanisms of formation, and mitigation research in the major food categories. It is an update on the first CIAA review paper, "A Review of Acrylamide: An Industry Perspective on Research, Analysis, Formation and Control." Initial difficulties with the establishment of reliable analytical methods, in most cases, have now been overcome, but challenges remain in terms of the need to develop simple and rapid test methods and certified reference materials. Many trials have been conducted under laboratory and experimental conditions in a variety of foods, and a number of possible measures have been identified to relatively lower the amounts of acrylamide in food. Promising applications were studied in reconstituted potato models by addition of amino acids or use of asparaginase. In bakery wares, predictive models have been established to determine the role of ammonium carbonate and invert sugar in acrylamide formation. Studies in several commercial foods showed that acrylamide is not stable over time in roasted and ground coffee. Some progress in relatively lowering acrylamide in certain food categories has been achieved, but at this stage can only be considered marginal. Any options that are chosen to reduce acrylamide in commercial products must be technologically feasible and must not adversely affect the quality and safety of the final product. PMID- 15759747 TI - The HEATOX Project. AB - HEATOX is the acronym for a European Union-funded project entitled Heat-Generated Food Toxicants: Identification, Characterization, and Risk Minimization. Acrylamide will be the main experimental focus, but identification of unknown toxicants in heated carbohydrate-rich foods will also be attempted. The project includes research on formation chemistry, food technology, analytical methods, hazard characterization, and exposure assessment. The results will finally be used in risk assessment and risk management advice. PMID- 15759748 TI - Overview of acrylamide monitoring databases. AB - Since high acrylamide levels in carbohydrate-rich food were reported in 2002, many research activities were started in order to gain knowledge on occurrence, formation, and prevention of this compound in food products. Among them, monitoring programs were conducted in many countries worldwide by official bodies as well as by the food industry. National and international bodies set up monitoring databases. In 2003, both the European Commission and the World Health Organization posted calls for data and placed their spreadsheets for the submission of data on the Web. The goal of the databases is to collect data for a reliable estimation of the exposure of consumers to acrylamide via the food chain. This paper describes the assessment of the data quality and outlines the composition of the data in the 2 databases, to date. PMID- 15759749 TI - Reduction of exposure to acrylamide: achievements, potential of optimization, and problems encountered from the perspectives of a Swiss enforcement laboratory. AB - The most important initiatives taken in Switzerland to reduce exposure of consumers to acrylamide are the separate sale of potatoes low in reducing sugars for roasting and frying, the optimization of the raw material and preparation of french fries, and campaigns to implement suitable preparation methods in the gastronomy and homes. Industry works on improving a range of other products. Although these measures can reduce high exposures by some 80%, they have little effect on the background exposure resulting from coffee, bread, and numerous other products for which no substantial improvement is in sight. At this stage, improvements should be achieved by supporting voluntary activity rather than legal limits. Committed and consistent risk communication is key, and the support of improvements presupposes innovative approaches. PMID- 15759750 TI - Acrylamide formation in food: a mechanistic perspective. AB - Earliest reports on the origin of acrylamide in food have confirmed asparagine as the main amino acid responsible for its formation. Available evidence suggests that sugars and other carbonyl compounds play a specific role in the decarboxylation process of asparagine, a necessary step in the generation of acrylamide. It has been proposed that Schiff base intermediate formed between asparagine and the sugar provides a low energy alternative to the decarboxylation from the intact Amadori product through generation and decomposition of oxazolidin-5-one intermediate, leading to the formation of a relatively stable azomethine ylide. Literature data indicate the propensity of such protonated ylides to undergo irreversible 1,2-prototropic shift and produce, in this case, decarboxylated Schiff bases which can easily rearrange into corresponding Amadori products. Decarboxylated Amadori products can either undergo the well known beta elimination process initiated by the sugar moiety to produce 3-aminopropanamide and 1-deoxyglucosone or undergo 1,2-elimination initiated by the amino acid moiety to directly generate acrylamide. On the other hand, the Schiff intermediate can either hydrolyze and release 3-aminopropanamide or similarly undergo amino acid initiated 1,2-elimination to directly form acrylamide. Other thermolytic pathways to acrylamide--considered marginal at this stage--via the Strecker aldehyde, acrolein, and acrylic acid, are also addressed. Despite significant progress in the understanding of the mechanistic aspects of acrylamide formation, concrete evidence for the role of the different proposed intermediates in foods is still lacking. PMID- 15759751 TI - Studies on the stability of acrylamide in food during storage. AB - Acrylamide levels in a variety of food samples were analyzed before and after 3 months of storage at 10 degrees-12 degrees C. The analysis was performed by liquid chromatography tandem mass spectrometry (LC/MS/MS) using deuterium-labeled acrylamide as internal standard. Acrylamide was stable in most matrixes (cookies, cornflakes, crispbread, raw sugar, potato crisps, peanuts) over time. However, slight decreases were determined for dietary biscuits (83-89%) and for licorice confection (82%). For coffee and cacao powder, a significant decrease occurred during storage for 3 or 6 months, respectively. Acrylamide concentrations dropped from 305 to 210 microg/kg in coffee and from 265 to 180 microg/kg in cacao powder. On the contrary, acrylamide remained stable in soluble coffee as well as in coffee substitutes. Reactions of acrylamide with SH group-containing substances were assumed as the cause for acrylamide degradation in coffee and cacao. Spiking experiments with acrylamide revealed that acrylamide concentrations remained stable in baby food, cola, and beer; however, recovery levels dropped in milk powder (71%), sulfurized apricot (53%), and cacao powder (17%). These observations suggest that variations in the acrylamide content of food, especially in coffee and cacao, can vary depending on the storage time because special food constituents and/or reaction products can affect the levels. PMID- 15759752 TI - Analytical methods used to measure acrylamide concentrations in foods. AB - The state-of-the-art of analysis for acrylamide in food is reviewed. The majority of analytical methods adopts a similar approach: addition of internal standard to the specimen, extraction with water, purification of extract using a solid-phase extraction cartridge, and then determination using either gas chromatography coupled to mass spectroscopy (GC/MS) after bromination, or direct measurement with liquid chromatography coupled to mass spectroscopy (LC/MS). The available methods generally show good agreement and are likely to be accurate. However, improvements in precision (within-laboratory) and repeatability (between laboratory) are needed by particular data users. PMID- 15759753 TI - Acrylamide analysis: assessment of results from six rounds of Food Analysis Performance Assessment Scheme (FAPAS) proficiency testing. AB - Six proficiency tests have now been completed in an ongoing program of the UK Food Analysis Performance Assessment Scheme (FAPAS) for the analysis of acrylamide in a range of food matrixes. Homogeneous test material samples were requested by laboratories throughout the world, with 29 to 45 submitting results for each test. Results were analyzed by appropriate statistical procedures, and z scores were awarded for reported values. In the absence of both legislation and collaborative trial data, the target standard deviation was derived from the Horwitz equation, although it is acknowledged that there is a need to establish a "fit for purpose" target standard deviation specifically for acrylamide analysis. Participants were encouraged to use the analytical method routinely used in their own laboratory and to provide details of their procedure. Close examination of the data submitted indicates that performance is generally acceptable in terms of accuracy. There is no significant difference between results submitted by gas chromatography and liquid chromatography (GC and LC) methods, and no method dependency on the use of internal standards or sample size. However, choice of extraction solvent may be important, with indications that plain water is an acceptable extraction method. There is evidence from the most recent test that direct (underivatized) GC methodology may present problems, but more data are required and this aspect will be monitored in the continuing proficiency testing program. PMID- 15759754 TI - Results from two interlaboratory comparison tests organized in Germany and at the EU level for analysis of acrylamide in food. AB - After the publication of high levels of acrylamide (AA) in food, many research activities started all over the world in order to determine the occurrence and the concentration of this substance in various types of food. As no validated methods were available at that time, interlaboratory studies on the determination of AA in food were of the highest priority. Under the boundary conditions of applying well-established evaluation schemes, the results of 2 studies conducted by the Federal Institute for Risk Assessment (BfR) in Germany and by the European Commission's Directorate General Joint Research Center (JRC) exhibited an overall acceptable performance of the participants in these studies. Nevertheless, many laboratories showed problems in determining AA in food with a complex matrix such as cocoa. The results of analysis also showed a broader variation of AA for samples with low AA concentrations and indicated a bias of the results obtained by gas chromatography-mass spectrometry without derivatization. Improvements of the performance of some laboratories appeared to be necessary. PMID- 15759755 TI - Committee on Natural Toxins and Food Allergens. Marine and freshwater toxins. AB - There have been major developments this past year in the Marine and Freshwater Toxins topic area (formerly Phycotoxins). These include AOAC approval and inauguration of a new AOAC Presidential Task Force on Marine and Freshwater Toxins to accelerate methods validation, and the appointment of several new Topic Advisors. A joint FAO/IOC/WHO group addressing biotoxins in molluscan bivalves is also relevant to this report and to the new Task Force. The AOAC Presidential Task Force on Marine and Freshwater Toxins is an international group that, in late November 2004, consisted of 90 world experts and stakeholders. Chaired by this General Referee, the group establishes methods priorities based on analytical methods criteria, determines fitness for purpose, identifies and reviews available methodologies, recommends methodologies for validation, and identifies complementary analytical tools. Once appropriate analytical methodology has been identified or developed, the Task Force is able to identify financial and technical resources necessary to validate the methods. The first two formal meetings of the Task Force were held in Bethesda, MD, on May 19, 2004 and in St. Louis, MO, on September 22, 2004. These meetings were held in conjunction with the XI International IUPAC Symposium on Mycotoxins and Phycotoxins and the 118th AOAC INTERNATIONAL Annual Meeting and Exposition, respectively. The Bethesda meeting served to introduce members of the group to the AOAC Community/Task Force model and to discuss objectives, concerns, general workings, and communications. The meeting concluded on an encouraging note, with a commitment from AOAC to help provide financial resources for the review of nonproprietary methods deemed high priority by the Task Force. This development was seen as an important step toward reaching methods validation objectives. The terms of reference for the Task Force were approved by the AOAC Board of Directors in late June, 2004. They described the Task Force membership as composed of voting and nonvoting members, with the voting members consisting of 13 members (12 plus the Chair). Voting members comprise of a balance of government regulators, academics, and industry members. No single agency has more than 2 voting members. Task Force members serve as experts in the field and agree to identify other experts; recommend individuals who can serve on the Task Force and as Chair; develop and prioritize a list of marine and freshwater toxins that need validated methods; assist in identifying existing methods for validation through AOAC validation programs; and recommend to the AOAC INTERNATIONAL Board of Directors policies and procedures necessary to accomplish the mission of the Task Force. They endeavor to actively support the work of the Task Force through garnering of sources of funding (except where prohibited by employer); identifying potential participating laboratories, sample identification and acquisition; and increasing program awareness among stakeholders. They assist AOAC in the identification of study directors and in the development of quality measurement tools by participating in the validation of methods and by identifying venues for members of the Task Group or the community to gather and assist with meeting content. Prior to the September 2004, AOAC Annual Meeting, the Task Force approved a set of Analytical Methods Selection Criteria, which are critical to the mission of the Task Force. They can be found, along with the Terms of Reference, roster of members, and other information, on the Task Force Web site at http://www.aoac.org/marine toxins/task_force.htm. The September 22, 2004 Task Force meeting in St. Louis included discussion of 2 interlaboratory studies, a proprietary kit for domoic acid by enzyme-linked immunosorbent assay (ELISA; Biosense Labs AS, Bergen, Norway) and also a nonproprietary liquid chromatography (LC) method for paralytic shellfish poisoning (PSP) toxins by precolumn oxidation (James F. Lawrence, Health Canada). These 2 methods were recommended by the Task Force for review by AOAC in September 2004. The group also discussed future priority directions, aspects of interlaboratory studies and official methods of analysis, other methods validation issues, future meetings, and funding. In addition to the Task Force meeting, 2 subgroup meetings were held. One subgroup addressed strategies to replace the mouse bioassay for brevetoxins with alternative modern methods based on ELISA or LC/mass spectrometry (MS). Brevetoxin metabolites, toxicity issues, and extraction conditions as well as future field studies were addressed in detail. The receptor binding assay (RBA)/saxitoxins subgroup addressed several aspects of the methodology, radiolabeled saxitoxin, and comparisons of mouse bioassay and RBA response. Both subgroups were productive and were seen as very useful by the participants. Task Force attendees generally agreed that subgroups are the most effective means of progressing towards validation of new methods and of ensuring thorough discussions of methods under consideration. By the time of their next meeting (April 2005) at the "Marine and Freshwater Toxins Analysis: 1st Joint Symposium and AOAC Task Force Meeting" in Baiona, Spain, the Task Force will have several well developed new subgroups in the areas of okadaic acid and dinophysis toxins, yessotoxins, domoic acids, and ciguatoxins. Some of the subgroups will hold face-to-face meetings in Spain and others will meet at future symposia or joint meetings. It is likely that training sessions will be associated with multiple Task Force meetings planned for 2005. Details on these meetings can be found on the Task Force Web site. Although the Task Force has experienced rapid growth, the addition of new members to the group, especially industry and government stakeholders, is encouraged. Task Force member Michael Quilliam, NRC Canada, provided the information given below on a joint CODEX group of special relevance to the new Task Force. This group met in late September 2004. For more information, see http://www.who.int/foodsafety/chem/meetings/biotoxin/en/. PMID- 15759756 TI - Committee on Natural Toxins and Food Allergens. Mycotoxins. PMID- 15759757 TI - Committee on Food Nutrition. Fat-soluble vitamins. PMID- 15759758 TI - Committee on Residues and Related Topics. Pesticides and other chemical contaminants. AB - One new topic needs to be added to this refereeship this year. This topic is Multiresidue Methods for Pesticides in Foods by GC/MS and LC/MS/MS. Last year the Synthetic Pyrethroids method was declared Final Action with comments. The comments were "Change applicability statement to: This method is applicable for the determination of 8 synthetic pyrethroid pesticides in wheat, oranges, and tomatoes at 0.10-2.0 mg/kg." The topic Determination of Residues of Triazines and Their Chloro-Metabolites in Raw Agricultural Commodities has now been filled. The topic Dioxins by GC/MS remains vacant. PMID- 15759759 TI - Committee on Residues and Related Topics. Metals and other elements. AB - This is the 12th report of the General Referee covering developments during the past year, presenting selected reports and concluding with recommendations for consideration by the Methods Committee on Residues and Related Topics. PMID- 15759760 TI - Committee on Microbiology and Extraneous Materials. Food microbiology, non-dairy. PMID- 15759761 TI - Efficacy testing of disinfectants. AB - The U.S. Environmental Protection Agency's (EPA) Office of Pesticide Programs (OPP) has the responsibility for regulating antimicrobial products, including sporicides, used to treat and decontaminate inanimate surfaces. In response to the anthrax (Bacillus anthracis) attacks of 2001 and the associated need for verifying the performance of chemicals for building decontamination, the EPA initiated research in late 2003 to evaluate and improve efficacy test methods for sporicides. The OPP Microbiology Laboratory located at the Environmental Science Center, Ft. Meade, MD is the lead laboratory. Through funding provided by EPA's Office of Research and Development (Safe Buildings Program), a collaborative research plan has been established to address several key issues. Research is currently being conducted on 2 fronts: (1) the evaluation of quantitative methodology for assessing the efficacy of sporicides, and (2) the development and comparative testing of selected modifications to improve the AOAC Sporicidal Activity Test (AOAC Method 966.04). Future studies will include the evaluation of candidate surrogates of B. anthracis using a quantitative method, and a multilaboratory validation study of a quantitative method-surrogate combination. The General Referee is serving as the Principal Investigator for all research described in this report, and has the overall responsibility for the technical conduct of the projects. In cases where the General Referee has oversight of projects that involve official collaborative studies and validation support from AOAC INTERNATIONAL, AOAC officials and the Committee Chair will determine the appropriate mechanism for formal study review. The 2003 General Referee report provides the background on the development and direction of the research projects. The preliminary data, general conclusions, next steps, and recommendations are provided in this report. PMID- 15759762 TI - Committee on Additives, Beverages, and Food Process-Related Analytes. PMID- 15759763 TI - Committee on Food Nutrition. PMID- 15759764 TI - Safety Committee. PMID- 15759765 TI - Tour de life of a CIO. Cyclists and CIOs need similar qualities to complete the course. PMID- 15759766 TI - Bar coding and RFID. PMID- 15759767 TI - Disease management. PMID- 15759768 TI - Electronic health record. PMID- 15759769 TI - Emergency preparedness. PMID- 15759770 TI - Integrating PACS. PMID- 15759771 TI - IT and biomedical devices. PMID- 15759772 TI - Patient-centric portals. PMID- 15759773 TI - Regional networks. PMID- 15759774 TI - Telehealth. PMID- 15759775 TI - The future is in the cards. Building continues on the foundation Peter Velez has formed for patient-controlled medical records. PMID- 15759776 TI - Nursing informatics of tomorrow. One of nurses' new roles will be agents of change in the healthcare revolution. PMID- 15759777 TI - Nurturing IT credibility. Think of yourself as a seller of ideas. PMID- 15759778 TI - Wireless attractions. The devices sell themselves when introduced in a physician friendly package. PMID- 15759779 TI - EMRs in nursing facilities. Are they different from those in acute-care facilities? PMID- 15759780 TI - Revenue cycle management. Hospital leaders see opportunities and needs in a crucial area. PMID- 15759781 TI - Disposing of idle technology. Using an above-reproach method restores resources and preserves peace of mind. PMID- 15759782 TI - Joining the HIT revolution. Should you buy or lease the tools you need? PMID- 15759783 TI - Maximizing security standards. Technology can help achieve more than HIPAA requires. PMID- 15759784 TI - [Comparative study between rapid urease test, imprint and histopathological study for Helicobacter pylori diagnosis]. AB - Helicobacter pylori infection is relevant, due to its role in different gastric pathologies; there are several diagnostic methods, which are divided into invasive and non-invasive. In the first category histology has been considered the gold standard, while other methods include imprint with Gram stain and rapid urease test, which can produce dependable results. The aim of this study is to compare several available techniques for H. pylori diagnosis to know their sensitivity and specificity. We studied 88 patients, 50 women and 38 men, with age range from 17 to 83 years (48.8+/-14.3) from the Endoscopy Department of the 20 de Noviembre Medical Center; in all five to eight biopsies were taken from gastric mucosa to perform rapid urease test, Gram-stained imprint and histology. Endoscopic findings included gastritis (87.50%), pangastritis (2.30%) and another diagnoses (10.22%). Rapid urease test was positive in 40 patients and negative in 48, while imprint was 34 and 54, respectively. Sensitivity and specificity were as follows: rapid urease test, 84.8 and 78.5%, and imprint 75.8 and 83.6%, respectively. In conclusion, H. pylori diagnosis by rapid urease test is a dependable and quick method. On the other hand, imprint is useful but depends on the ability of the personnel who perform the test. PMID- 15759785 TI - [Self-expandable prostheses for palliation of gastric cancer: report of four cases]. AB - Self-expandable prostheses have gained in popularity for palliation tumors of the gastrointestinal tract. We present four cases of patients with gastric adenocarcinoma who were palliated by self-expandable metallic prostheses. Two patients had undergone total gastrectomy with esophago-jejunal anastomosis. In one there was recurrent cancer in the anastomosis and the other had non-malignant stenosis. The remaining two patients had unresectable tumors. Correct placement of prostheses was possible in all cases; however, maintenance of adequate hydration and acceptable nutritional status per orem was possible only in three of four patients. In all cases there was temporary slight pain, in one, slight hemorrhage, and in one the prosthesis migrated and it was necessary to withdraw it surgically. Although these are only a few cases, we can say that self expandable prostheses are a promising alternative to palliate gastric cancer. PMID- 15759786 TI - [Complications of the management of secondary peritonitis with contained-open abdomen. Comparison of the Bogota's bag vs polypropylene mesh]. AB - BACKGROUND: Surgery is the cornerstone regarding the treatment of secondary peritonitis, although the complications of the contained-open abdomen are common, and are usually severe. OBJECTIVES: To identify the complications with the use of the "Bogota's bag" compared with polypropylene mesh in contained-open abdomen. To identify other possible morbidity and mortality risk factors, independently of the mesh type usage. METHODS: One hundred patients with secondary peritonitis were included in this retrospective cohort study. They were treated from January 2000 to March 2002. Fifty subjects were operated on with the "Bogota's bag " (50%) and 50 with the polypropylene mesh (50%). Mannheim's peritonitis severity index, complications and mortality using central tendency and dispersion means, the logistic regression analysis, prevalence and prevalence-ratios were analyzed. RESULTS: Most frequent etiologies of secondary peritonitis were intestinal anastomosis dehiscence, hollow viscera perforation, and severe pancreatitis. Observed complications were: perforation and intestinal fistula in 13 cases (13%); 10 (20%) out of these belonged to the polypropylene mesh group, and 3 (6%) to the Bogota's bag group (p = 0.036). Evisceration was observed in 8 cases (8%); 2 (4%) in the polypropylene mesh group, and 6 (12%) in the Bogota's bag group (p = 0.140). Hemorrhage was present in 30 cases (30%); 16 (32%) in the polypropylene mesh group, and 14 (28%) in the Bogota's bag group (p = 0.662). Colonization was present in 24 cases (24%); and this was mainly evident in the polypropylene mesh group (p = 0.019). Mortality reached 42% (42 cases), and was associated to postoperative bleeding (p = 0.004), peritonitis' severity (p = 0.0012), and surgical reinterventions. CONCLUSIONS: The contained-open abdomen is the best surgical option to treat infection and the compartmental abdominal syndrome in secondary peritonitis, however, complications may be relevant. These results demonstrated a direct relationship between peritonitis severity, the need for surgical re-interventions and mortality. The use of polypropylene mesh was related to an increased risk of intestinal fistulization and prothesis-associated infection. PMID- 15759788 TI - [Gastrointestinal tuberculosis]. AB - INTRODUCTION: Intestinal tuberculosis usually is not contemplated as a possible cause of chronic abdominal pain; nevertheless, incidence of this disease is increasing even in well-developed countries. MATERIALS AND METHODS: Four patients with intestinal tuberculosis who required surgical treatment. RESULTS: All patients had a history of chronic abdominal pain, presented acute complications of the disease, and required surgery. Three patients had AIDS and the remaining patient had chronic renal failure. CONCLUSIONS: Intestinal tuberculosis is a disease that should be considered as possible cause of chronic abdominal pain in patients with altered immunology. PMID- 15759787 TI - [Variation of intraabdominal pressure caused by abdominoplasty in healthy women]. AB - OBJECTIVE: To study pressure changes of the abdominal compartment during abdominoplasty in healthy women. PATIENTS AND METHODS: A prospective cohort of 14 women between 30 and 57 years old, who underwent abdominoplasty for aesthetic purposes wer included in this study. All patients were free of any associated medical disorder with a body mass index less than 30 kg/m2. The surgical procedure consisted of the plication of internal borders of the abdominal anterior rectal muscles with non absorbable sutures. Lipectomies were performed according to the flap shape and were sutured in two planes. Abdominal pressure was indirectly measured through the urinary bladder (Kron's technique). before the surgical procedure, after the plication of the aponeurotic muscle plane and after the adipocutaneous flap closure. A full written agreement was obtained from all patients before the surgical procedure. RESULTS: Average age was 39.5+/-8.06 years. The anterior rectal muscle diastasis was 6.2+/-2.17 cm and the plicature 8.0+/-2.0 cm. The basal pressure was 0.1+/-0.3 mm Hg, whereas, after the plication it increased to 4.9+/-0. 7 mm Hg. An intraabdominal pressure was of 6.56+/-1.1 mm Hg was found after the adipocutaneous closure the A significant statistical difference (p < or = 0.001) was found. We did not observe any pulmonary complication during the first 30 days following the surgical procedure. CONCLUSIONS: In our series, the closure of the adipocutaneous flaps increased the intraabdominal pressure 31%, but no pulmonary complication was observed during the follow-up period. Since intraabdominal pressure above 10 mm Hg induces hemodynamic changes and lung restriction, the increment produced after the adipocutaneous closure should be considered to prevent morbidity associated to intraabdominal hypertension. PMID- 15759789 TI - [Dysphagia secondary to lingual thyroid]. AB - BACKGROUND: Lingual thyroid is a rare developmental anomaly originating from aberrant embryogenesis during descent of the gland from its site of origin, the foramen caecum, to its eutopic location. OBJECTIVE: We present the case of a 30 year-old female with a history of dysphagia and pharyngeal foreign body sensation over the past 12 months, associated with hypothyroidism. METHODS: Diagnostic and therapeutic approaches are discussed. RESULTS: Diagnosis of lingual thyroid was made and transoral thyroidectomy was performed as procedure-of-choice. Microscopic analysis revealed normal thyroid tissue with abundant colloid. The patient had an uneventful recovery and was discharged 2 days after surgery. Gamma gram with Tc99m was scheduled for postoperative follow-up. CONCLUSIONS: Transoral lingual thyroidectomy followed by life-long hormonal therapy is the appropiate approach for patients with severe symptoms due to lingual thyroid hypertrophy associated with hypothyroidism. PMID- 15759790 TI - [Cholesterol ester storage disease]. AB - The goal of this paper is to present a clinical case of a 4 year old boy, with hepatomegaly, splenomegaly and intestinal lipid infiltration due to a inborn error of lipid metabolism known as cholesterol ester storage disease. The main clinical manifestations were hepatomegaly, splenomegaly, hypertriglyceridemia, hypercholesterolemia. Duodenal endoscopy showed a yellow appearance of the mucous, and the histological study revealed the presence of macrophages with granular material. Liver biopsy showed steatosis infiltration at the hepatocytes, and macrophages with lipids. This disease is due to a lisosomal acid lipase partial deficiency, that is a glicoprotein that metabolize the hydrolysis of ester of cholesterol and triglycerides. The name of this pathology is cholesterol ester storage disease, but when the deficiency is total the name is Wolman's disease. We conclude that in all the children whit a clinical picture of hepatomegaly, splenomegaly, hypertriglyceridemia and hypercholesterolemia it is obligatory to rule out an inborn error of lipid metabolism like Wolman's disease or cholesterol ester storage disease. PMID- 15759791 TI - [Pathogenic molecular mechanisms in non-alcoholic steatohepatitis]. AB - Non-alcoholic steatohepatitis is a chronic disease that occurs in persons without significative consumption of alcohol, characterized by macrovesicular steatosis, mixed inflammatory infiltrate, and diverse degrees of fibrosis. It can progress to cirrhosis and its evolution to hepatocellular carcinoma has been described. It principally occurs in patients with obesity, diabetes mellitus, and hyperlipidemia, and is at present considered a manifestation of metabolic syndrome with insulin resistance. In pathogenesis, diverse factors, fundamentally insulin resistance as a mechanism that determines hepatic steatosis, have been described. Later, alteration of signalling cascades, oxidative stress, and other mechanisms occur that lead to inflammation, necrosis, and finally to hepatic fibrosis, the details of which will be described in this review. PMID- 15759792 TI - [Pseudomembrane colitis-related toxic megacolon]. PMID- 15759793 TI - [Choledocholithiasis associated to Dormia basket impactation]. PMID- 15759794 TI - [Importance of Helicobacter pylori diagnosis]. PMID- 15759795 TI - Diseases and partial mortality in Montastraea annularis species complex in reefs with differing environmental conditions (NW Caribbean and Gulf of Mexico). AB - We documented the prevalence of diseases, syndromes and partial mortality in colonies of the Montastraea annularis species complex on 3 reefs, and tested the assumption that a higher prevalence of these parameters occurs when reefs are closer to point-sources of pollution. One reef was isolated from the impact of local factors with the exception of fishing, 1 potentially influenced by local industrial pollutants, and 1 influenced by local urban pollution. Two reefs were surveyed in 1996 and again in 2001 and 1 in 1998 and again in 2001. In 2001, colonies on all reefs had a high prevalence of the yellow-band syndrome and a relatively high degree of recent partial mortality, while the prevalence of black band and white-plague diseases was low although a new sign, that we named the thin dark line, had relatively high prevalence in all reefs. As no direct relationship was found between disease prevalence and local environmental quality, our results open the possibility that regional and/or global factors may already be playing an important role in the prevalence of coral disease in the Caribbean, and contradict the theory that coral disease prevalence is primarily related to local environmental degradation. Reasons that may partially explain these findings are the high level of potential pathogen connectivity within the Caribbean as a result of its circulation patterns coupled to the large land derived pollutants and pathogens input into this Mediterranean sea, together with the surface water warming effects which stress corals and enhance pathogen activity. PMID- 15759796 TI - Diseases and causes of mortality among sea turtles stranded in the Canary Islands, Spain (1998-2001). AB - This paper lists the pathological findings and causes of mortality of 93 sea turtles (88 Caretta caretta, 3 Chelonia mydas, and 2 Dermochelys coriacea) stranded on the coasts of the Canary Islands between January 1998 and December 2001. Of these, 25 (26.88%) had died of spontaneous diseases including different types of pneumonia, hepatitis, meningitis, septicemic processes and neoplasm. However, 65 turtles (69.89%) had died from lesions associated with human activities such as boat-strike injuries (23.66%), entanglement in derelict fishing nets (24.73%), ingestion of hooks and monofilament lines (19.35%), and crude oil ingestion (2.15%). Traumatic ulcerative skin lesions were the most common gross lesions, occurring in 39.78% of turtles examined, and being associated with Aeromonas hydrophila, Vibrio alginolyticus and Staphylococcus spp. infections. Pulmonary edema (15.05%), granulomatous pneumonia (12.90%) and exudative bronchopneumonia (7.53%) were the most frequently detected respiratory lesions. Different histological types of nephritis included chronic interstitial nephritis, granulomatous nephritis and perinephric abscesses, affecting 13 turtles (13.98%). Ulcerative and fibrinous esophagitis and traumatic esophageal perforation were the most frequently observed lesions in the esophagus, being associated in the majority of the cases with ingestion of fishing hooks. Larval nematodes of the Anisakidae family caused gastritis in 15 turtles (16.13%). Necrotizing and/or granulomatous hepatitis were the lesions most commonly observed in the liver (27.95%). Traumatic lesions included necrotizing myositis (10.75%) mainly caused by entanglement in fishing nets or boat-strikes, and amputation of 1 or 2 flippers (25.81%) by netting. Traumatic erosions and/or fractures of the carapace/plastron mainly caused by boat-strikes were also observed (26.88%). Eye lesions included heterophilic keratoconjunctivitis, ulcerative keratitis and heterophilic scleritis, affecting 7 turtles (7.53%). PMID- 15759797 TI - Picoeucaryot alga infecting blue mussel Mytilus edulis in southern Norway. AB - During summer 2001, blue mussels Mytilus edulis with abnormal shell growth were collected near Kragero, southern Norway. The mussels had green spots in their mantle tissues, mainly posteriorly and ventrally, and in the adductor muscle. Mussels from 4 sites had a prevalence of green spots varying from 2 to 71% that correlated well with shell deformities. Histological examination revealed the presence of round or ovoid algae, 0.9 to 1.5 x 1.2 to 2.4 microm, free within haemocytes and in the lesions, characterised by an inflammatory response and the presence of cellular debris. The alga contain a relatively large nucleus, 1 chloroplast and 1 mitochondrion. Size and morphology suggest that the alga might be a picoeucaryot green alga. Infection of mussel tissues appears to start in the posterior mantle edge, near the siphons, and spread anterior-ventrally in the mantle connective and storage tissues-occasionally spots were also found in the gonad follicles. Large infected areas were also observed in sinuses within the adductor muscle. Only mussels that were 3 yr old or more were infected. Deformations apparently resulted from years of continuous shell formation by a contracted, partly deformed mantle. Most deformed mussels had eroded shells, allowing some light penetration through the exposed, thin nacre. Young, thin shelled mussels were not infected. The present work suggests that the alga has, at least partially, a parasitic relationship with the mussels, and is associated with pathological alterations in mussel tissues. PMID- 15759798 TI - Herpes virus in juvenile Pacific oysters Crassostrea gigas from Tomales Bay, California, coincides with summer mortality episodes. AB - Pacific Crassostrea gigas and eastern C. virginica oysters were examined between June 2002 and April 2003 from 8 locations along the east, west and south USA coasts for oyster herpes virus (OsHV) infections using the A primer set in a previously developed PCR test. Only surviving Pacific oysters from a mortality event in Tomales Bay, California, USA, where annual losses of oysters have occurred each summer since 1993, were infected with a herpes-like virus in 2002. PCR examination using template amounts of both 50 and 500 ng were essential for OsHV detection. Sequence analysis indicated that the Tomales Bay OsHV was similar to that identified in France with the exception of a single base pair substitution in a 917 bp fragment of the viral genome. However, unlike the French OsHV-1, the Tomales Bay OsHV did not amplify with the primer pair of a second OsHV-1 PCR assay, suggesting that further characterization of these viruses is warranted. No evidence of Cowdry type A viral infections characteristic of herpes virus infections or other pathogens were observed in OsHV-infected oysters. Hemocytosis, diapedesis and hemocyte degeneration characterized by nuclear pycnosis and fragmentation were observed in infected oysters, which is consistent with previous observations of OsHV infections in France. Together these data suggest that OsHV may be associated with the annual summer Pacific oyster seed mortality observed in Tomales Bay, but establishment of a causal relationship warrants further investigation. PMID- 15759799 TI - Characteristics of inhibition of infectious pancreatic necrosis virus (IPNV) by normal rainbow trout Oncorhynchus mykiss serum. AB - We studied the characteristics of rainbow trout serum (RTS) inhibitory activity against infectious pancreatic necrosis virus (IPNV). Serum inhibition was related to the serum source and host cell in which the virus had been propagated. IPNV was more efficiently inhibited by RTS in salmonid cell lines than in non-salmonid cell lines, with inhibition highest in rainbow trout gonad (RTG)-2 cells. The RTS sensitivity of the virus was modified by the cell line through which the virus passed, with multiple passages through Chinook salmon embryo (CHSE)-214 cells producing a virus that was less sensitive to RTS. The RTS inhibition level was dependent on cell density: at a cell density of < or = 2 x 10(5) cells ml(-1), inhibition was insignificant (tissue culture infective dose 50% = 10(-1.1) TCID50 ml(-1) reduction); however, above a density of 3 x 10(5) cells ml(-1), the inhibition level was very high (> or = 10(-6.3) TCID50 ml(-1) reduction). The salmonid sera tested showed high inhibition, except for brook trout serum (BTS), while non-salmonid sera did not inhibit IPNV, replication on RTG-2 cells. Pretreatment of cultured cells with RTS prior to exposure did not affect inhibition of IPNV and thus did not mask a viral receptor. The RTS inhibition level was dependent on the time of serum addition, with inhibition being maintained for at least 16 h postinfection. Pretreatment of IPNV revealed that the virus is directly inhibited by RTS, and more strongly so when RTS is present during viral replication. PMID- 15759800 TI - Production of monoclonal antibodies against grouper nervous necrosis virus (GNNV) and development of an antigen capture ELISA. AB - Five (2 IgG, 3 IgM) monoclonal antibodies (MAbs) against the G9508KS strain of grouper nervous necrosis virus (GNNV) were produced and characterized. All 5 MAbs showed positive signals in the retina of GNNV-infected grouper larvae and in the cytoplasm of GNNV-infected GF-1 cells using immunohistochemistry staining. Two MAbs reacted with the denatured capsid protein derived from GNNV-infected GF-1 cells in Western blot analysis, but did not react with the GNNV recombinant capsid protein expressed by E. coli in an indirect immnunosorbent assay (ELISA). All 5 MAbs were able to neutralize GNNV, tiger puffer NNV (TPNNV) and barfin flounder NNV (BFNNV), while only 2 of the MAbs neutralized striped jack NNV (SJNNV). A capture ELISA system based on the use of MAbs for capture and a rabbit polyclonal antibody for detection was developed. When absorbance values higher than 0.5 were judged to be positive, the sensitivity of the capture ELISA system was 2.5 ng per well of purified GNNV protein or 6.5 x 10(4) TCID50 per well of GNNV supernatant from culture cells. This capture ELISA system could become a more specific and sensitive tool for NNV diagnosis in the field and in routine laboratories. PMID- 15759801 TI - Effect of Pseudomonas sp. MT5 baths on Flavobacterium columnare infection of rainbow trout and on microbial diversity on fish skin and gills. AB - Use of Pseudomonas sp. strain MT5 to prevent and treat Flavobacterium columnare infection was studied in 2 experiments with fingerling rainbow trout Oncorhynchus mykiss. In the first experiment, length heterogeneity analysis of PCR-amplified DNA fragments (LH-PCR) was used to assess the effect of antagonistic baths on the microbial diversity of healthy and experimentally infected fish. In the 148 samples studied, no difference was found between bathed and unbathed fish, and 3 fragment lengths were detected most frequently: 500 (in 75.7% of the samples), 523 (62.2%) and 517 bp (40.5%). The species contributing to these fragment sizes were Pseudomonas sp., Rhodococcus sp. and F. columnare, respectively. A specific PCR for detection of Pseudomonas sp. MT5 was designed, but none of the tissue samples were found to be positive, most likely indicating poor adhesion of the strain during bathing. LH-PCR was found to be a more powerful tool for detecting F. columnare in fish tissue than traditional culture methods (chi2 = 3.9, df = 1, p < 0.05). Antagonistic baths had no effect on the outbreak of infection or on fish mortality. F. columnare was also detected in healthy fish prior to and after experimental infection, indicating that these fish were carriers of the disease. In the second experiment, intensive Pseudomonas sp. MT5 antagonistic baths were given daily to rainbow trout suffering from a natural columnaris infection. Again, the antagonistic bacteria had no effect on fish mortality, which reached 95 % in both control and antagonist-treated groups in 7 d. PMID- 15759802 TI - Effects of fish age versus size on the development of whirling disease in rainbow trout. AB - We examined the effects of both fish age and size on the development of resistance to whirling disease in Erwin strain rainbow trout. Previously, we demonstrated that juvenile rainbow trout became resistant to development of the disease when first exposed to triactinomyxons of the parasite Myxobolus cerebralis at about 9 wk post-hatch when raised at 12 degrees C, but ages and sizes of fish used in that experiment were confounded (Ryce EKN, Zale AV, MacConnell E [2004] Dis Aquat Org 59:225-233). In this study, rainbow trout of the same age and different sizes, and the same size and different ages, were exposed to the parasite to distinguish the influences of age and size. Fish were reared at 3 different water temperatures prior to exposure to produce groups with different growth rates and were exposed to the parasite at 7 or 9 wk post-hatch. Disease severity was affected by both age and size at first exposure, but the effects were not independent. An increase in fork length from 36 to 40 mm among fish exposed at 7 wk post-hatch did not confer increased resistance, but the same increase in size at 9 wk post-hatch did. Similarly, an increase in age from 7 to 9 wk post-hatch among fish exposed at 36 mm fork length did not confer increased resistance, but the same increase in age at 40 mm did. Rainbow trout must be both 9 wk post-hatch or older and at least 40 mm in fork length at time of exposure to exhibit enhanced resistance to whirling disease. Resistance to disease was not associated with the level of skeletal ossification. PMID- 15759803 TI - Joint effects of parasitism and pollution on oxidative stress biomarkers in yellow perch Perca flavescens. AB - Yellow perch Perca flavescens were collected from a contaminated site and a reference site in the St. Lawrence River, Quebec, Canada. Fish were assessed for oxidative stress (lipid peroxidation and reduced glutathione levels) and parasitism by the nematode Raphidascaris acus and metacercariae of the digenean Apophallus brevis. Lipid peroxidation is not only considered a measure of oxidative stress, but of stress in general, and thus serves as an indicator of fish health. Fish from the contaminated site exhibited higher levels of lipid peroxidation than those from the reference site, independent of parasitic infections. However, fish infected with R. acus at the contaminated site tended to have higher levels of lipid peroxidation than uninfected fish at the same site, whereas no difference was observed between infected and uninfected fish at the reference site. Yellow perch infected with > 10 metacercariae of A. brevis expressed higher levels of lipid peroxidation than those infected with < 10 metacercariae at both the contaminated and the reference sites. No differences were found in levels of reduced glutathione in liver or muscle in relation to site or either parasite species. Results support the use of lipid peroxidation as a biomarker of water contamination. They further suggest that lipid peroxidation may be used as a biomarker of pathological effects caused by parasitism. Most importantly, results demonstrate that contaminants and parasites occurring together exacerbate oxidative stress in fish, suggesting that parasitized fish in polluted environments are in a poorer state of health than uninfected fish. PMID- 15759804 TI - Antiserum to the gp116 glycoprotein of yellow head virus neutralizes infectivity in primary lymphoid organ cells of Penaeus monodon. AB - Yellow head virus (YHV) is an invertebrate nidovirus that has caused mass mortality of cultured Penaeus monodon in Asia. In this study, we investigated whether mouse polyclonal antiserum raised against the YHV gp116 or gp64 structural glycoproteins could neutralize YHV infectivity as determined using an in vitro quantal assay in primary cultures of lymphoid organ cells. Anti-gp116 antiserum showed virus-neutralizing activity whereas anti-gp64 antiserum failed to inhibit infection. The results suggest that gpl16 antiserum blocks binding of virions to cellular receptors to facilitate YHV entry into lymphoid organ cells. PMID- 15759805 TI - Bacterial clearance rate and a new differential hemocyte staining method to assess immunostimulant activity in shrimp. AB - New methods were developed to assess immunostimulant efficacy in the black tiger shrimp Penaeus monodon. Test shrimp were fed with 2 or 4 % yeast extract (YE) coated feed while controls were fed non-coated feed. After 4 wk of feeding, individual shrimp were assessed for total hemocyte counts (THC), the number of granular hemocytes (GH) and rate of bacterial clearance. For hemocyte counts, formalin-fixed hemolymph was stained with 1.2 % Rose Bengal in 50 % ethanol for 20 min at room temperature. Some of this mixture was used for THC with a hemocytometer while some was smeared on a microscope slide and left to dry before counterstaining with hematoxylin for GH counts. By this technique, high quality smears were obtained for accurate differential counts. Bacterial clearance assays were used to assess the sum effect of humoral and cellular defense mechanisms. Vibrio harveyi was injected intramuscularly at 1 x 10(8) cells per shrimp and hemolymph was collected in anticoagulant at 0, 15, 30 and 60 min post-injection for quadruplicate drop counts (20 microl) on TCBS agar. Total hemocyte counts for shrimp fed with 4 % YE were significantly higher (p < 0.05) than those for shrimp fed with non-coated feed. The percentage of granular cells and the rates of bacterial clearance for the YE-fed shrimp were higher than those for shrimp fed the control diet. These 2 methods provide a simple and rapid comparison of shrimp groups for differences in anti-bacterial defense capacity. PMID- 15759806 TI - [Greater life expectancy: effect on life insurance]. PMID- 15759807 TI - [The natural history of congenital defects and deformities of the spine (II)]. PMID- 15759808 TI - [Chronic musculoskeletal pain--aspects for assessment in private disability compensation (II)]. PMID- 15759810 TI - [Traumatic partial amputation of a penis--a reconstruction of the circumstances of the accident]. AB - A 61-year-old man was admitted to hospital with a partially severed penis. The head of the penis (glans) had been completely severed, and the skin of the shaft and the corpora cavernosa had been ripped open. In the hospital the patient reported that his penis got caught in the hose attachment of an old Kobold vacuum cleaner that he was using to inflate an air mattress. He later made contradictory statements in his report to the insurance company, so we were asked to reconstruct the circumstances of the accident. The literature available to us only makes clinical observations about similar accidents, always with the assumption that the vacuum cleaner was used during masturbation or in order to achieve an erection. According to our reconstruction of the accident and an investigation of the vacuum cleaner attachment, however, we could not rule out the possibility of a household accident as described by the patient. PMID- 15759809 TI - [MRI--diagnosis bone bruise]. AB - The diagnosis bone bruise was at first established by means of MRI in 1987. Up to now the discussion as to whether bone bruise has to be interpretated simply as a radiological finding or a harbinger of post-traumatic arthritis is controversial. In this article, we demonstrate two different cases in individual medical assessments of bone bruise with different patterns of injuries. While regarding the literature concerning bone bruise, we discuss the consequences of these injuries in IMA. PMID- 15759811 TI - [Medical opinions in court--a challenge for communication between physicians and lawyers]. AB - Medical opinions in court constitute a great challenge in regard to communication between physicians and lawyers. Considerable differences in their professional training and requirements frequently result in misunderstandings. Therefore physicians and lawyers need to learn more about each other and interact when evaluating opinions. A synergistic rather than a conflicting approach would help judges to find the happy medium between accepting medical reports unreservedly and coming to their own conclusions in more or less total disregard of the medical opinion presented in court. The expert, on the other hand, must ensure that the judge will understand the opinion as it was meant. Neither of them can call upon a translator. PMID- 15759812 TI - [21st ICLAM--Congress of the International Committee for Life, Disability and Health Assurance Medicine]. PMID- 15759813 TI - Carotid atherosclerosis: is it treatable? AB - The assessment of brachial-artery FMD is a reliable, reproducible and noninvasive tool for evaluating endothelial dysfunction, which, together with endothelial inflammation, causes carotid atherosclerosis. Atorvastatin therapy can reverse endothelial dysfunction after 4 months, and may have significant anti inflammatory effects if continued for more than 4 months. Importantly, statin therapy can be used to treat carotid atherosclerosis. PMID- 15759814 TI - The controversy over diagnosis and treatment of facet pain. PMID- 15759815 TI - Changes in flow-mediated dilatation, cytokines and carotid arterial stenosis during aggressive atorvastatin treatment in normocholesterolemic patients. AB - BACKGROUND: Aggressive statin therapy to reduce low-density lipoprotein (LDL) cholesterol in patients with normal LDL-cholesterol levels reduces the incidence of future cardiovascular events and enhances atherosclerotic regression in the common carotid artery. We tried to quantify changes in flow-mediated dilatation (FMD), inflammatory cytokines, and the severity of carotid arterial stenosis, after aggressive statin administration to patients with normal LDL-cholesterol levels, and stroke or transient ischemic attack ipsilateral to carotid arterial stenosis. METHODS: Twenty patients with at least a 50% reduction in the diameter of the carotid artery were studied. Atorvastatin 10 mg daily was prescribed for 4 months. Serial changes in the severity of carotid arterial stenosis, and brachial artery FMD were evaluated by high-resolution ultrasound. Tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), vascular cell adhesion molecule (VCAM), intracellular adhesion molecule (ICAM), soluble endothelin-1 (ET-1), and high-sensitivity C-reactive protein (hs-CRP), were measured before, and after 2 and 4 months of, atorvastatin treatment. RESULTS: Atorvastatin significantly decreased plasma levels of total cholesterol and LDL cholesterol after 1, 2 and 4 months of treatment (p < 0.01). FMD showed significant improvement after only 4 months (10.0% vs 6.8% at baseline; p = 0.019). The overall severity of carotid arterial lesions was not significantly reduced by atorvastatin. Changes in TNF-alpha, IL-1beta, IL-6 and ET-1 showed trends towards a progressive decline after atorvastatin, but none of the cytokines was reduced significantly. FMD did not correlate with the severity of carotid arterial stenosis, lipid profile, or cytokine levels. CONCLUSION: Atorvastatin effectively reduced plasma concentrations of total cholesterol and LDL-cholesterol, and had beneficial effects on endothelial function, in Chinese patients with carotid arterial stenosis and normal LDL-cholesterol levels. PMID- 15759816 TI - Lumbar zygapophyseal joint injections in patients with chronic lower back pain. AB - BACKGROUND: This study was designed to assess the diagnostic value and clinical benefits of lumbar zygapophyseal joint injections in patients with chronic lower back pain. METHODS: Two hundred and seventy-seven patients (136 males and 141 females, aged 15-82 years) with chronic lower back pain were enrolled in the trial and met the following criteria: pain for more than 1 year; no root signs; and no history of back surgery. Under fluoroscope, a 0.8-1.5 mL mixture of lidocaine, betamethasone dipropionate and iopamidol (1:1:0.5) was injected into each joint after intra-articular localization of the needle tip was confirmed. A questionnaire with a pain scale was administered immediately or the day after injection, and then after 1, 3, 6 and 12 weeks. Partial arthrograms were reviewed by a radiologist. RESULTS: Four hundred and forty-nine joint injections were performed in 277 patients (L3-4, n = 76; L4-5, n = 272; L5-S1, n = 101). Bilateral injections were performed in 117 patients (42.2%). The study group comprised 204 patients (73.6%) with an excellent or good response, whereas the control group comprised the remaining 73 patients (26.4%). The rates of good response in the study group were 72.1% (147/204) after 3 weeks, 40.7% (83/204) after 6 weeks, and 31.4% (64/204) after 12 weeks. Partial arthrograms revealed 25 patients (9.0%) with synovial cysts (L3-4, n = 3; L4-5, n = 14; L5-S1, n = 8); 23 of these patients (92.0%) had a good response to the injections. Five of the 6 patients with spondylolysis (83.3%), having abnormal communication between the injected and contiguous joint, had a good response to the injections. The abovementioned, abnormal partial-arthrogram findings correlated significantly with the rate of good response to the injections. Although 3 patients had contrast medium extravasated into the epidural space during injection, none of the 277 patients had deteriorating lower back pain after the injections. CONCLUSION: Lumbar zygapophyseal joint injections, as a useful diagnostic tool for facet joint syndrome, could also have useful palliative effects in the management of chronic lower back pain. PMID- 15759818 TI - Treatment of nonunion of humeral shaft fracture with dynamic compression plate and cancellous bone graft. AB - BACKGROUND: This study was conducted to evaluate the treatment of aseptic nonunion of the humeral shaft with a dynamic compression plate (DCP) and cancellous bone graft. METHODS: One hundred and five cases of nonunion of a humeral shaft fracture between 1982 and 2001 were analyzed retrospectively. The study population comprised 66 males and 39 females with an average age of 46.2 years (range, 17-81 years). Sixty-seven fractures were defined as atrophic nonunion, and 20 as hypertrophic nonunion, whereas 18 could not be defined clearly. All the fractures were managed by open reduction and internal fixation with DCP and cancellous bone graft. The mean follow-up period was 20 months (range, 14-28 months). RESULTS: All nonunion fractures united within an average of 16 weeks (range, 10-26 weeks). Complications included 4 patients with temporary radial-nerve palsies, and 3 patients with wound infections. At the final follow-up, shoulder and elbow functions of the operated limbs were all satisfactory. CONCLUSION: Fixation by DCP with supplemental cancellous bone graft is a reliable and effective treatment for nonunion of a humeral shaft fracture. PMID- 15759817 TI - A randomized, controlled, clinical study of thymosin alpha-1 versus interferon alpha in [corrected] patients with chronic hepatitis B lacking HBeAg in China [corrected]. AB - BACKGROUND: This study was designed to compare the efficacy and safety of thymosin-alphal (T-alpha1) with that of interferon-alpha (IFN-alpha) in patients with chronic hepatitis B who were positive for hepatitis B virus (HBV) DNA and hepatitis B envelope antibody (anti-HBe). METHODS: Fifty-six patients were randomly divided into groups A and B. Both groups were comparable (p > 0.05) at baseline regarding age, sex, and alanine aminotransferase (ALT) levels. Group A patients received T-alpha1 1.6 mg subcutaneously twice weekly, while group B patients received IFN-alpha 5 million IU daily for 15 days, then thrice weekly for 6 months. Results from the 2 groups were compared with data from a group of 30 patients never treated with IFN-alpha and who were followed-up for 12 months (historical control [HC] group); the 3 groups were comparable (p > 0.05). RESULTS: After treatment, a complete response (ALT normalization and HBV DNA loss) occurred in 8 of 26 patients in group A (30.8%) and 14 of 30 in group B (46.7%; chi2 = 1.476, p = 0.224). After a follow-up period of 6 months, a complete response was observed in 11 of 26 patients in group A (42.3%) and 7 of 30 in group B (23.3%; chi2 = 2.299, p = 0.129). The rate of complete response was significantly greater in the IFN-alpha than HC group at the end of therapy (46.7% vs 3.3%; chi2 = 15.022, p = 0.0001), and in the T-alphal than HC group at the end of follow-up (42.3% vs 3.3%; chi2 = 12.566, p = 0.0001). Ten of the 12 T-alphal responders (i.e. partial responders; 83.3%) experienced sustained, non-detectable HBV DNA after 6 months' treatment; 6 of the 14 T-alphal non-responders (42.9%) showed a delayed response of non-detectable HBV DNA during the follow-up period. Corresponding values for group B patients were 50% (9/18) and 0% (0/12). The rate of delayed response was significantly higher in group A than the other 2 groups (chi2 = 6.686, p = 0.010; chi2 = 4.964, p = 0.038), whereas the rate of flare was higher in group B than in the other 2 groups (chi2 = 3.445, p = 0.063; chi2 = 7.668, p = 0.006), during the follow-up period. Unlike IFN-alpha, T-alphal was well tolerated, i.e. no adverse effects were noted in group A. CONCLUSION: These results suggest that a 6-month course of T-alpha1 therapy is effective and safe in patients with anti-HBe-positive chronic hepatitis B; T-alpha1 can reduce HBV replication in such patients. Compared with IFN-alpha, T-alpha1 is better tolerated and seems to induce a gradual and more sustained normalization of ALT and loss of HBV DNA. Combination therapy with T-alpha1 and IFN-alpha or nucleoside analogs for hepatitis B warrants further study. PMID- 15759819 TI - Reversible electroencephalographic and single photon emission computed tomography abnormalities in Hashimoto's encephalopathy. AB - Hashimoto's encephalopathy (HE) is a well-established disease that occurs most commonly in patients with euthyroidism. We report 3 patients diagnosed with HE. Cases 1 and 2 were young females who had experienced intractable seizures and presented with cognitive impairment. Reversible slow waves on the electroencephalogram (EEG), and nearly complete recovery of cognition, were noted after steroid treatment. Case 3 was an elderly male who presented with a sudden onset of vertigo, diplopia, and recurrent gait ataxia. Reversible hypoperfusion in the upper portion of the left cerebellum, on (99m)Tc hexamethylpropyleneamine oxime (HMPAO) single photon emission computed tomography (SPECT), was noted after steroid treatment. We also review 25 cases from the literature of patients diagnosed with HE. All patients had antithyroid antibodies. A reversible inflammatory process is supported by brain magnetic resonance imaging, SPECT, EEG and cerebrospinal fluid studies. Vasculitis is the most probable pathogenesis according to laboratory findings. Steroids are the treatment of choice, and result in a favorable outcome when administered early in the disease course. PMID- 15759820 TI - Severe hypoglycemic coma due to insulin autoimmune syndrome. AB - Insulin autoimmune syndrome, characterized by the presence of insulin-binding autoantibodies and fasting or late postprandial hypoglycemia, is a rare cause of hypoglycemia. We report a patient with pulmonary tuberculosis who developed recurrent spontaneously post-absorptive hyperinsulinemic hypoglycemia after treatment with anti-tuberculous drugs. Imaging studies of the pancreas were unremarkable, and selective intra-arterial calcium stimulation with hepatic venous sampling for insulin failed to show a gradient, thus almost completely excluding the possibility of occult insulinoma or nesidioblastosis. Examinations of sera, however, disclosed a high titer of polyclonal insulin-binding autoantibodies containing at least 2 classes of binding sites, 1 with high affinity but low capacity, and the other with low affinity but high capacity. An oral glucose tolerance test revealed high serum levels of total insulin associated with relatively low levels of free insulin, but not of C-peptide, suggesting binding of the released insulin to autoantibodies. Regretfully, shortly after the withdrawal of isoniazid, the patient died of respiratory failure unrelated to hypoglycemia, and whether these antibodies were induced by isoniazid remains unknown. We recommend that insulin autoimmune syndrome be one of the differential diagnoses in patients with hyperinsulinemic hypoglycemia. PMID- 15759821 TI - Synchronous parathyroid and papillary thyroid carcinoma. AB - Concomitant thyroid disease is not unusual among patients with primary hyperparathyroidism. However, the simultaneous occurrence of parathyroid and thyroid carcinoma is extremely rare. We report a 38-year-old man with primary hyperparathyroidism who presented with osteitis fibrosa cystica complicated with pathologic femoral neck fracture. Preoperative investigation for exclusion of multiple endocrine neoplasia did not find evidence of medullary thyroid carcinoma or pheochromocytoma, but imaging studies revealed the presence of nodules in the right lobe and a parathyroid lesion over the left inferior pole of the thyroid gland. Total thyroidectomy, left parathyroidectomy, and bipolar hemiarthroplasty of the left hip were then performed simultaneously. The resected specimens were pathologically identified as papillary thyroid carcinoma and parathyroid carcinoma, respectively. After the operation, 131I ablation therapy was administered at a dose of 120 mCi. Additional doses of 30 mCi were given yearly as serum thyroglobulin level became elevated. Serum calcium level remained normal during yearly follow-up. Although parathyroid carcinoma is an uncommon cause of parathyroid hormone-dependent hypercalcemia, it should nonetheless be given due consideration because its surgical approach differs from that of parathyroid adenoma. As the coexistence of parathyroid and non-medullary thyroid carcinoma has previously been reported, the possibility of both malignancies must also be considered in the setting of primary hyperparathyroidism with thyroid nodules. If confirmed with preoperative parathyroid scintigraphic and other laboratory studies, an optimal outcome may be achieved with complete resection of both tumors at the time of initial operation, followed by adjunctive therapy. PMID- 15759822 TI - Association of AIDS and bipolar mania with rapid progression to dementia and death. AB - Neuropsychiatric complications of human immunodeficiency virus (HIV) infection or acquired immune deficiency syndrome (AIDS) may present clinically as acute or chronic organic brain syndrome, or mimic functional psychiatric diseases. Among such psychiatric diseases, mania tends to occur with increased frequency after the onset of AIDS. We report a case in which manic manifestations were noted before the diagnosis of AIDS. The patient had no past or family history of mood disorders, but had risk factors for HIV infection. He had a rapid downhill course from initial manic symptoms to depression, dementia and then death within 10 months. Such rapid cognitive deterioration into AIDS dementia after mania is consistent with previous reports. Cases like this will become more common with spread of the AIDS pandemic in Asian regions, including Taiwan. Clinicians should be mindful of HIV infection/AIDS as a differential diagnosis in patients with manic episodes and risk factors for HIV infection. PMID- 15759823 TI - Intrapelvic migration of a Kirschner wire. AB - Intrapelvic migration of a Kirschner wire, used in the treatment of hip fracture, is rare. We report a case of intrapelvic migration of a Kirschner wire 10 years after its placement. Even though the patient was asymptomatic, we proceeded to remove the migrated wire using laparoscopic techniques. Given the increasing number of hip operations being performed, and given that intrapelvic foreign bodies can result in serious or fatal complications, it is important to keep this problem in mind in patients who have had previous hip surgery. PMID- 15759824 TI - The practices of drug companies marketing the same drug on the human and veterinary markets. PMID- 15759825 TI - Difficult for private practitioners to obtain reference material. PMID- 15759826 TI - An ethicist's commentary on over-vaccination. PMID- 15759827 TI - Animal health laboratory reports--cattle. Bovine abortion update, 1998-2004. PMID- 15759828 TI - Prevalence of Giardia and Cryptosporidium in beef cows in southern Ontario and in beef calves in southern British Columbia. AB - In 1998 and 1999, fecal samples were collected from 669 beef cows on 39 farms located within 10 counties of Ontario. Overall prevalences of Giardia, Cryptosporidium muris, and Cryptosporidium parvum in cows were 8.7%, 10.6%, and 18.4%, respectively. Of the 39 farms sampled, Giardia was detected on 64%, Cr. muris on 72%, and Cr. parvum on 90%. Cryptosporidium parvum was detected in 28% of the cows in 1998 and in 5.2% in 1999. Differences between the 2 y were attributed to sampling during calving in 1998 and during gestation in 1999. In 1998, Giardia, Cr. muris, and Cr. parvum were detected in herds provided with municipal water. In 1998, 193 calves were sampled from 10 farms, representing 4 watersheds, in British Columbia. Thirty-six percent of the calves exhibited signs of diarrhea. Overall prevalences of Giardia and Cryptosporidium spp. in calves were 36% and 13%, respectively. There was evidence that calves with Giardia were more likely to develop scours. Restricting cattle from surface water during periods of high shedding may reduce watershed contamination. PMID- 15759829 TI - Seroprevalence of antibodies against bovine leukemia virus, bovine viral diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in dairy cattle in Saskatchewan. AB - Blood was drawn from 1530 dairy cows in 51 herds. For antibodies against bovine leukemia virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum, 37.4%, 2.7%, and 5.6% of cows were test positive, respectively, while 29.2% of herds had unvaccinated animals with > or = 1:64 for bovine viral diarrhea virus. PMID- 15759830 TI - Polioencephalomalacia in adult sheep grazing pastures with prostrate pigweed. AB - Polioencephalomalacia was diagnosed in 2 animals from different farms. In apparently healthy animals from same farms, fecal thiaminase and a significant reduction in erythrocyte trans-ketolase activity was observed. The presence of thiaminase in Amaranthus blitoides could have contributed to the development of polioencephalomalacia in sheep grazing on natural pastures. PMID- 15759831 TI - Ovarian remnant syndrome in a 5-year-old bitch. AB - A 5-year-old German shepherd bitch was presented with a 3-week history of vaginal bleeding. Ovarian remnant syndrome was diagnosed on vaginoscopy, vaginal cytology, serum progesterone levels, ultrasonography, and exploratory laparotomy. The condition resolved following surgical excision of the remaining ovarian tissue. PMID- 15759832 TI - Perspectives on emerging zoonotic disease research and capacity building in Canada. AB - Zoonoses are fundamental determinants of community health. Preventing, identifying and managing these infections must be a central public health focus. Most current zoonoses research focuses on the interface of the pathogen and the clinically ill person, emphasizing microbial detection, mechanisms of pathogenicity and clinical intervention strategies, rather than examining the causes of emergence, persistence and spread of new zoonoses. There are gaps in the understanding of the animal determinants of emergence and the capacity to train highly qualified individuals; these are major obstacles to preventing new disease threats. The ability to predict the emergence of zoonoses and their resulting public health and societal impacts are hindered when insufficient effort is devoted to understanding zoonotic disease epidemiology, and when zoonoses are not examined in a manner that yields fundamental insight into their origin and spread. Emerging infectious disease research should rest on four pillars: enhanced communications across disciplinary and agency boundaries; the assessment and development of surveillance and disease detection tools; the examination of linkages between animal health determinants of human health outcomes; and finally, cross-disciplinary training and research. A national strategy to predict, prevent and manage emerging diseases must have a prominent and explicit role for veterinary and biological researchers. An integrated health approach would provide decision makers with a firmer foundation from which to build evidence-based disease prevention and control plans that involve complex human/animal/environmental systems, and would serve as the foundation to train and support the new cadre of individuals ultimately needed to maintain and apply research capacity in this area. PMID- 15759833 TI - Bile duct carcinoma and nasal adenocarcinoma in a guinea pig. AB - A guinea pig with a low grade bile duct carcinoma and nasal papillary adenocarcinoma also had a collagen nevus or lipoma that was considered insignificant in the overall management of the case. The bile duct carcinoma was treated successfully through surgery. The clinical presentation of the adenocarcinoma, which was not treated, is described. PMID- 15759834 TI - Leadership in animal welfare: the role of veterinary colleges. PMID- 15759835 TI - Associate compensation: self-employed or salaried? PMID- 15759836 TI - Diagnostic ophthalmology. Orbital neoplasm (primary or secondary). PMID- 15759837 TI - [Fluorescence angiography of the anterior eye segment in the evaluation of impaired microcirculation within the diagnosis of severity of eye burns]. AB - We used fluorescence angiography to study the vascular microcirculation in the anterior eye segment of 14 patients with severe eye bums. The severity of eye burns is not only related with an impaired blood circulation in the conjunctiva, it also depends on a microcirculation status of episcleral vessels. Different degrees of permeability of fluorescein through the conjunctival or episcleral vessels make it possible to specify circulation disorders in both cases. Whether the impaired microcirculation in the anterior eye segment is transient or not can be established through a second fluorescence angiography made during days 7-8 from the burn trauma. The severity of eye burn depends directly on the sizes of ischemic regions of the sclera. PMID- 15759838 TI - [A comparative evaluation of astigmatic neutrality of 5 mm self-sealing tunnel incision of different types]. AB - The impact produced by incisions of 2 examined types on the corneal refraction was studied in 40 patients (40 eyes) shared between 2 groups according to an incision type. 5 mm tunnel self-sealing incisions of 2 types, i.e. temporal corneal one longer by 3 mm, and upper linear sclerocorneal one stabilized by horizontal suture, were made use of. The obtained refraction was evaluated on week 1 and then in 1 and 3 months after surgery. Group 1 (surgical approach: 5 mm linear sclerocorneal tunnel incision): the mean value of induced astigmatism made 0.68 D during the initial 3 postoperative days, 0.62 D in 1 month after surgery and 0.73 D in 3 months after surgery. Group 2 (surgical approach: 5 mm temporal corneal tunnel incision): the above value made 1.1 D during the initial 3 postoperative days, 0.7 D in one month after surgery and 0.52 D in 3 months. PMID- 15759839 TI - [Results of primary IOL implantation after extraction of congenital cataract in two-year babies]. AB - The results of aspiration of congenital cataract and those of IOL implantation were analyzed for 15 children (17 eyes) aged 2 years or less. The specificity of the postoperative clinical course after implantation of different IOL types and of changes of refraction as well as peculiarities of the growth of the aphakic eye in unilateral cataract versus its match were analyzed. Exudative reaction, stages I-II, is the main problem encountered in the early postoperative period irrespective of a type of the implanted IOL. Implantation of rigid IOLs is associated with a higher risk of development of secondary cataract in the late postoperative period. The postoperative changes of refraction do not depend on an IOL type. The artiphakic eye becomes myopic as the anatomic-optic parameters of the growing eye change. There are no reliable differences between the growth indices in the anterior-posterior axis of the artiphakic and growing eyes. PMID- 15759840 TI - [Secondary cataract and IOL type and the shape of its optic part]. AB - Although cortical remains are purged from the capsule sac by special techniques and better IOL models are used for implantation, opacification of the posterior capsule (OPC) persists to be one of the most frequent complications in modern cataract surgery. The frequency rate of OPC depends on used evaluation criteria. A total of 165 eyes were examined after cataract extraction for OPC. Four types of IOL were used; they are hydrophobic acryl three-part IOL (AcrySof MAZOVA, n = 46), hydrophobic acryl all-in-one IOL (ArySofSA30AL, n = 38) and silicone three part (CeeOn911A, Pharmacia, n = 42). OPC were evaluated by EPCO 2000 computer soft. When IOL with the straight edge of the optic axis were in use (both silicone and acryl ones), there was registered, in 1 year after surgery (p < or = 0.05) a reliably lower frequency and intensity of OPC versus IOL with the curved edge of the optic axis. The shaped curvature in the posterior capsule encompassing tightly the IOL edge prevented further migration of lens cells. The IOL material (hydrophobic acryl or silicone) is of no importance for the OPC development. There was no reduction in visual acuity and no laser capsulotomy was made in any of the groups of patients. PMID- 15759841 TI - [A subjective evaluation of epibulbar and etrobulbar anesthesia in cataract phacoemulsification]. AB - Described in the paper are comparative descriptions of the efficiency of epibulbar and retrobulbar anesthesia made through evaluating the sensation of pain experienced by patients in cataract phacoemulsification and IOL implantation. The patients (1046 subjects, aged 28 to 89) were shared between 2 groups; they received an identical premedication by midazolam, peroral administration, 3.75 mg in 1 h before surgery. Epibulbar anesthesia with 1% proximetacaine hydrochloride, administered via instillator, was used in Group 1 (524 patients). Analgesia with 2% lidocaine hydrochloride solution (injections, 1:1) was made in Group 2 (522 patients). The subjective pain sensations experienced by patients during subsequent surgical stages were evaluated by an international 6-point scale. The pain sensations were found to vary from 0 to 4 points (mean point value--2.25) in Group 1. The most severe pain was experienced by the patients during the introduction of the phacoemulsificator tip into the anterior chamber of the eye (mean pain value--1.75 points) as well as during IOL implantation (mean pain value--1.25). The instillator-based analgesia does not normally provoke any complications and is well tolerated by patients; it can be recommended for cataract phacoemulsification. PMID- 15759842 TI - [Principles of background stimulation in the treatment of amblyopia]. AB - Described in the paper are the results of a comparative evaluation of treatment of amblyopia by computer techniques based on the modern understanding of the mechanisms of impaired visual functions, as observed in amblyopia, comprising stimulation of different canals of the visual system, which were affected or not. Better clinical-and-functional results were fixed in patients with high amblyopia by using the "Ambcar" soft stimulating the magnocellular system by sinusoidal grates with the eyes being fixed on moving objects as well as in patients with moderate amblyopia by using the "eYe" and "Krestiki" soft tools affecting different canals of the visual system and different levels of the parvocellular system. Amblyopia should be treated with respect to individual differences between affected functions of the visual system in each patient, and a therapy method should be chosen in proportion to a disease severity. PMID- 15759843 TI - [Functional diagnostics of retinal ischemia: Muller cells and neovascularization of the retina in diabetic retinopathy]. AB - Electrography was made for 58 patients (110 eyes) with proliferative diabetic retinopathy distinguished by neovascularization. Fluorescence angiography and electroretinography were dynamically made for 1-1.5 years in 18 patients with preproliferative diabetic retinopathy. The hyperfunction of Muller cells with an increasing glial index, which was detected earlier by us in progressing retinal ischemia, was also observed in retinal neovascularization. The regular monitoring of the dynamics of electroretinogram (ERG, made several times) makes it possible to evaluate in each separate case a degree of neovascularization risk. When the value of the glial index abruptly drops in a patient, who had earlier regular increases in the index value, there is a high probability of a fast development of retinal neovascularization. Regular ERG examinations are advisable for the evaluation of a neovascularization risk. PMID- 15759844 TI - [The specificity of the ultrastructural pattern of neoplastic vessels of the retina in diabetic retinopathy]. AB - The electron microscopy of 14 samples of epiretinal membranes obtained in the course of transciliary vitrectomy performed in proliferative diabetic retinopathy (PDR) showed a majority of neovascularized vessels to be represented by those of the venular type or capillaries. It can be concluded on the basis of examination results that the structures of neovascularized vessels can be highly diverse in diabetic retinopathy, which depends on a variety of factors: 1) proliferation stage; 2) division of the microcirculation bed; and 3) end of the epiretinal membrane where a given vessel is located as to the growth epicenter. Finally, described are both neo-vessels with a virtually normal structure of the vascular wall and atrophying vessels. The signs of vascular reduction grow to be more pronounced at later stages. PMID- 15759845 TI - [Adequacy of prescribing the disaggregants in vascular pathology of the eye fundus and in primary open-angle glaucoma]. AB - The functional activity of blood platelets was studied in 936 patients with vascular pathology of the eye fundus (central chorioretinal dystrophy--CCRD; preproliferative diabetic retinopathy--PPDR; proliferative diabetic retinopathy- PDR; hypertensive retinopathy--HP; thrombosis of retinal veins--TRV; and primary open angle glaucoma--POAG) by using 2 methods: the routine one, which does not expose the real process of platelets' aggregation in the blood bed, and a method, which is closer to the conditions in vivo. The non-physiological ("tube"-based) methods of determination of platelets' aggregation overstate almost 3-fold the share (%) of patients needing disaggregants. The disaggregation therapy is indicated in 28% of cases; it is more often prescribed for patients with TRV (52.2%), HP (40%), PPDR (35.5%), POAG (29%), PDR (21.5%) and CCRD (15.3%). An acute decrease of the aggregation ability of platelets was registered in 22.5% of the examinees: drugs intensifying the functional activity of platelets were used. The conclusion is that iatrogenic hemorrhages can be reduced by using adequate methods for the determination of the functional activity of platelets and by prescribing an adequate medicamental therapy. PMID- 15759846 TI - [An experimental action of new injection-scleroplasty substance on eye tissues]. AB - An experimental study was made with 8 eyes of chinchilla rabbits for the purpose of investigating the interplay between the allotransplant (administered in the Tenon's space of the eyes of experimental animals), suggested by us, on the one hand, and the neighboring ocular components, on the other hand. According to the results, the used substance is subject to necrolysis accompanied by an induced local inflammation response. As the substance dissolved, it was found to be displaced by connective tissue. At first, the tissue was granulated, then, it became more compact with its structure resembling the sclera; the tissue tightly abutted the latter intensifying its properties. It is noteworthy, that the inflammation zone was limited to the administration region. With due respect to the above, it can be stated that the suggested method can be clinically used in future as a full sclera-fixing procedure. PMID- 15759847 TI - [Quant therapy and the composition of the moister of the eye anterior chamber (an experimental study)]. AB - We examined 12 rabbits, 6 of whom (12 eyes) were exposed to magneto-infrared laser radiation (MILR) and another 6 (12 eyes) were controls. The parameters of pulse and continuous infrared LED radiation were as follows: wavelength--860 nm, pulse capacity--2 W, mean radiation capacity--10 mW, magnetic field strength--up to 17 mTl. A study of the moister of the anterior chamber showed a MILR-induced activated metabolism, i.e. a better acid-base balance (ABB), more intense oxygenation in the ocular tissues and decreased acidosis. Higher concentrations of buffer bases (ABEe and SBEc) cause shifts in ABB towards metabolic alkalosis. A lower concentration of glucose denotes intensified processes related with its utilization. A lack of changes in the quantity of salts in the moister of the anterior chamber rules out the possibility of that the content of glucose would go down due to its dissolution with a big volume of newly produced moister. A lack of an increase in the concentration of whole protein, as observed after MILR, can be regarded as indirect evidence to absence of any adverse effect on the vascular wall. PMID- 15759848 TI - [New regulatory protein isolated from the bovine eye lens and its action on the cataract development in rat in vitro]. AB - The regulatory protein was isolated from the eye lens extract by using an early designed scheme including by means of salting-out of proteins by ammonium sulphate, isoelectrofocusing in pH gradient and electrophoresis in PAAG. A high purity fraction of the regulatory protein was obtained. The localization of the regulatory protein in the rat-eye lens was investigated by means of primary rabbit antibodies obtained within the case study and by FITS-marked secondary antibodies. Cataractogenesis was induced, in vitro, in Wistar rat lenses through adding, to the cultivation medium, hydrogen peroxide (0.5 mM) or calcium chloride (15 mM). The regulatory protein isolated from the bovine eye lens was added alongside with damaging antibodies to the nutrition medium, concentration 10(-12) mg/ml. The lenses were cultivated for as long as 8 days at 37 degrees C. The degree of opacification of lenses was evaluated visually with the help of a lined substrate as well as by spectrophotometry. The studied protein was shown immunohistochemically to be localized in the intercellular space of the lens epithelium in the region of the basic membrane. The cataractogenesis-related research of the regulatory protein was made on rabbit eye lenses, which were cultivated as a whole for as long as 8 days in vitro. Their transparency and morphology were preserved in them in full since they were cultivated in a serum free nutrition without admixture of any destructive agents. Opacification of lenses was induced in vitro by changing the concentration of calcium ions in the cultivation medium or through adding hydrogen peroxide to the medium. The valuations of the lens opacity degree as observed in different research series and made by visual observation well correlate with the results of spectrophotometry of lenses made after their cultivation. It can be stated that the studied regulatory protein, when added to the cultivation medium, enhances about two-fold the lens transparency versus the lenses cultivated in the catactogenesis-containing medium. Finally, very small doses of the regulatory protein isolated from the bovine eye lens were found to prevent cataractogenesis in rats in vitro. Since the studied regulatory protein was localized by us in the region of epithelium, it can be suggested that its protective action is conditioned by its ability to contribute to regulating the main biological processes occurring in the lens capsule. PMID- 15759849 TI - [An experience of using electric milling cutter in the treatment of children with chronic dacryocystitis]. PMID- 15759850 TI - [A rare observation of chiasm apoplexy]. PMID- 15759851 TI - [A case of phacoemulsification made after a previously performed radial keratotomy (specificity of estimation of IOL optic power)]. PMID- 15759852 TI - [A spontaneous escape of gadfly larva through the conjunctiva of a child's eye]. PMID- 15759853 TI - [Posttraumatic hyphema: pathogenesis and treatment]. PMID- 15759855 TI - [Life devoted to fight against blindness (on the occasion of the 130th birthday anniversary of V. P. Filatov)]. PMID- 15759854 TI - [New treatment methods in pathologies of the choroid and retina including the use of subthreshold power of diode infrared laser radiation]. PMID- 15759856 TI - [Screen in vivo induced gene of Mycobacterium tuberculosis by IVIAT]. AB - In order to search new candidates of pharmaceutical target, in vivo induced antigen technology (IVIAT) was used to screen in vivo induced (ivi) genes of Mycobacterium tuberculosis (M. TB). Genomic expression library of M. TB was first constructed with an inducible plasmid pKK223-8; the titer of the library was 1.02 x 10(5) CFU. Sera from ten tuberculosis patients were pooled and absorbed against in vitro-grown M. TB and Escherichia coli, and used to probe the genomic expression library. 16 positive clones were identified by immunological screen, including 22 ORF: two encoding lipid metabolism proteins, five information pathways proteins, two PE/PPE proteins, six intermediary metabolism and respiration proteins, one cell wall and cell processes protein, four conserved hypothetical proteins and two conserved hypothetical proteins with an orthologue in Mycobacterium bovis. Parts of these genes can be used as candidates of pharmaceutical target because they may be relate with virulence. PMID- 15759857 TI - [Genetic polymorphism of mitochondrial DNA in coding region in 16 ethnic populations of Yunnan]. AB - The genetic polymorphism of mitochondrial DNA in the coding region in 16 ethnic populations of Yunnan was analyzed using PCR-RFLP in a total of 654 samples. Seventeen haplogroups were found, four of which were undefined haplogroups. Haplogroup distribution and Principal Component (PC) analysis showed that the ethnic groups descended from Bai-Yue tribe have B, F and M7 as the predominant haplogroups,which indicated their origination from southern China. Haplogroup A, D and N9 were predominant in the Mongolian ethnic group, reflecting their north originated characteristics. The groups descended from Di-Qiang tribe shared the predominant haplogroups with both the south and north originated groups,which demonstrated that they inherit the maternal characteristics from both southern and northern populations. There is genetic difference among the populations in the same ethnic group and is usually smaller than that among the ethnic groups from different ancient tribes, but not necessarily smaller than that among the groups from the same tribe. PMID- 15759858 TI - [Phenotype positioning on chromosomes in a patient with the syndrome of partial trisomy 7p21.2-->pter]. AB - By using the techniques of human chromosome G-banding, high resolution banding and fluorescence in situ hybridization (FISH), we investigated a patient with the karyotype of partial trisomy 7p21.2-->pter. Combining with the comparative review of the clinical data in 14 cases with partial trisomy 7p syndrome from reported literatures,we searched for the correlation between the karyotype and the phenotype and between the karyotype and the associated gene in the patients with partial trisomy 7p syndrome. The results indicated that the 7p21.2-->p22 is the critical segment of partial trisomy 7p syndrome. The phenotypes of the genital malformation and the dislocation of hip joint are associated with the duplication of 7p15. The cardiac anomalies is resulted from the dysfunctions of several genes on the long arm of chromosome 7. The gene associated with the craniosynostosis may locate on the region of 7p21.2-->p21.3. PMID- 15759859 TI - [Pedigree study of pathological myopia]. AB - 90 pedigrees including 1822 individuals were investigated and analyzed to find out the genetic mode of pathological myopia in Chinese population. 169 screened nuclear pedigrees from the total were divided into two groups according to mating mode, Affected * Normal or Normal * Normal. Simple segregation analysis on A * N and N * N pedigrees was performed respectively. The results showed that A * N pedigrees fit the autosomal dominant inheritance, with segregation ratio 0.6033 and sporadic proportion 13.8%, while N * N pedigrees fit autosomal recessive inheritance, with segregation ratio 0.235245 and sporadic proportion 16.3%, although autosomal dominant inheritance could not be rejected. In complex segregation analysis,SAGE-REGD software was used to fit several genetic model, including Mendelian inheritance (major gene, dominant, recessive, codominant) and non-Mendelian inheritance (non-transmitted, environment, general), and at last all Mendelian inheritances including major gene, dominant, recessive, codominant inheritance were accepted, while codominant inheritance with minimus AIC was best fitted. Our study manifests that pathological myopia in Chinese population fits autosomal dominant and autosomal recessive inheritance with certain sporadic proportion, which demonstrates the high genetic heterogeneity of pathological myopia. PMID- 15759860 TI - [Evidence of the unreduced diploid eggs generated from the diploid gynogenetic progeny of allotetraploid hybrids]. AB - Following activation by UV-irradiated sperms from scatter scale common carp and without the treatment for doubling the chromosome number, the eggs generated by the diploid gynogenetic progeny (G1) with 100 chromosomes,developed into diploid gynogenetic progeny (G2) with 100 chromosomes. Both the males and females of the tetraploids with 200 chromosomes were obtained from the offsprings (G1 x AT) produced by mating the eggs of G1 with the diploid sperms from the allotetraploid hybrids. The results provided the evidence that diploid G1 is able to produce diploid eggs. In the oogonia of diploid six-month old G2, only the chromosomes in the metaphase of mitosis were observed in all slides, no bivalent chromosome being found, suggesting that the oogonia in six-month old G2 were not mature enough for meiosis I. The presence of the chromosome spreads with 100, 200 and 380 chromosomes observed in the metaphase indicated that the diploid G2 had the potential ability to generate diploid eggs with the probable mechanism of pre meiotic endoreduplication. In contrast, the spermatocytes of G1 x AT allotetraploids had the normal chromosomal behavior with only 100 bivalents found in meiosis I. The one-year old G2 possessed the slowly developmental ovaries which stayed at the oogonium stage for a long time, in which no primary and mature oocyte was found, whereas both the females and males of one-year old G1 x AT allotetraploids had normal ovaries and testes which reached maturity and produced diploid eggs and diploid sperms, respectively. The formation of the diploid eggs generated from the diploid gynogenetic progeny makes it available to establish a diploid hybrid clonal line, and will become an important source of production of diploid eggs. In addition, the bisexual fertile G1 x AT allotetraploids enriched the types of the tetraploids. PMID- 15759861 TI - [Phylogeny of chinese catfishes inferred from mitochondrial cytochrome b sequences]. AB - The mitochondrial DNA cytochrome b gene was sequenced from 27 catfish species representing 11 families and 24 genera catfishes in China. Aligning with cytochrome b sequences of eight catfish species from North America and Africa retrieved from GenBank, and selecting Astyanax mexicanu, Cyprinus carpio, and Sardinops melanostictus as outgroups, we constructed a matrix of 38 DNA sequences. The phylogenetic trees were constructed by using Bayesian method and Maximum Parsimony (MP) method. The results showed that (1) there are three base pair deletions of mitochondrial cytochrome b gene compared with Characiformes, Cypriniformes, and Clupeiformes; (2) the representatives of Chinese catfish species form a monophyletic group; and (3) the molecular phylogenetic trees constructed with both methods suggest that the families Sisoridae, Akysidae and Amblycipitidae form a monophyletic group, and the families Clariidae, Schilbidae, Ariidae, Ictaluridae, Cranoglanididae, Pangasiidae, Siluridae, Claroteidae, and Bagridae also form a monophyletic group. The families Cranoglanididae from China and Ictaluridae from North America form a sister-group relationship, and the families Clariidae, Ictaluridae, Siluridae, and Sisoridae are obviously monophyletic groups. But the position of the family Plotosidae was not resolved by Bayesian analysis and maximum parsimony inference. PMID- 15759863 TI - [Analysis of polymorphism on exon 2 of SLA-DRB in Wuzhishan, Erhualian and Pietrain pigs with PCR-RFLP and PCR-SSCP]. AB - Exon 2 of SLA-DRB gene in three strains of pigs was genotyped by PCR-RFLP and PCR SSCP methods. The strains involved were Wuzhishan, Erhualian and Pietrain, and the numbers of them were 17, 28 and 28, respectively. After genotyping by PCR RFLP with Msp I, all pigs showed the same band pattern (143 bp/102 bp) named M. However, four kinds of band patterns were created by Rsa I, that were A:141 bp/93 bp/11 bp, B:111 bp/69 bp/54 bp/11 bp, C:180 bp/54 bp/11 bp and D:93 bp/48 bp/39 bp/54 bp/11 bp. There were different pattern types in various pigs, AA and BB in Wuzhishan pigs, AA, BB and AB in Erhualian pigs, AA, CC and BD in Pietrain pigs. In each strain, A was the dominant band pattern, its frequency in Wuzhishan, Erhualian and Pietrain pigs were 0.69, 0.73 and 0.82, respectively. There were no significant differences in the frequency of A band pattern among these three strains. After genotyping by PCR-SSCP, seven pattern types (alphaalpha, alphadelta, betabeta, gammagamma, alphagamma, deltadelta and betaepsilon) were observed in these three populations. There were alphaalpha, alphadelta and betabeta in Wuzhishan pigs, alphaalpha, gammagamma and alphagamma in Erhualian pigs, and alphaalpha, deltadelta, alphadelta, betaepsilon and betabeta in Pietrain pigs. In each breed, the frequency of a band pattern was the highest in all defined bands. The frequency of delta band pattern followed to alpha band pattern in the populations of Wuzhishan and Pietrain pigs, corresponding to that, the frequencies of alphadelta pattern type were maximal in these two populations. The frequencies of all pattern types analyzed by PCR-RFLP and PCR-SSCP were Hardy Weinberg Equilibrium in Erhualian populations; however, the same results did not appear in the other two populations. PMID- 15759862 TI - [Molecular cloning and character analysis of the mouse zinc finger protein gene Zfp474 exclusively expressed in testis and ovary]. AB - A novel mouse zinc finger protein gene that contains C2HC/C3H domain was first isolated by using a data-mining tool called Digital Differential Display (DDD) from the National Center for Biotechnology Information. The full-length cDNA of this transcript was deduced and further confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Total four exons of the mouse gene spaning a 29 869 bp genomic DNA sequence was mapped to chromosome 18D1. The cDNA encodes a novel protein of 347 amino acids and the protein contains four C2HC/C3H domains. Northern blot analyses revealed that Zfp474 mRNA was exclusively expressed in testis and ovary and had one transcript. We hypothesize that Zfp474 functions as a germ cell specific transcription factor that plays important roles in spermatid differentiation and oocyte development. PMID- 15759864 TI - Cloning and characterization of TOM7-like gene in wheat. AB - The cDNA encoding TOM7 (translocase of outer mitochondrial membrane subunit 7) like protein in wheat was cloned through RT-PCR, and its genomic DNA fragment was subsequently cloned. This gene was tentatively designated as TaTOM7. It has no intron in the coding region and its product possesses one hydrophobic trans membrane domain in the middle, and one hydrophilic domain in the N-terminal and C terminal domain, respectively. The amino acid composition in the trans-membrane domain of the TaTOM7 subunit is highly conserved among plants, animals and fungi. According to the phylogenetic tree, TOM7-like proteins from different species can be classified into three groups, representing plants, animals and fungi respectively. TaTOM7 is a single- or oligo-copy gene in the wheat genome, displaying different expression levels between Ms2 near-isogenic lines in some tissues, suggesting a role of Ms2 in its expression. TaTOM7 was mapped to chromosome group three of wheat. PMID- 15759865 TI - [Mapping QTLs controlling vascular bundle and panicle-related traits from Yuanjiang common wild rice (Oryza rufipogon Griff.)]. AB - An advanced backcross population (BC3F2), derived from the cross between Yuanjiang common wild rice as the donor parent and Teqing as recurrent parent, was used to map QTLs controlling the number of vascular bundle and panicle related traits from Yuanjiang common wild rice. Seven QTLs controlling the number of large vascular bundles in peduncle were detected on chromosome 1, 2, 3, 4, 7 and 10. Five QTLs for the number of small vascular bundles in peduncle were mapped on chromosome 1, 2, 3, 4 and 8. A total of 15 QTLs for the number of primary and secondary rachis branches and spikelets per panicle were identified on all chromosomes except chromosome 11 and 12. Most of alleles derived from O. rufipogon Griff. showed negative effect that reduced significantly vascular bundle, rachis branches and spikelets per panicle, suggesting that during the evolution from common wild rice to cultivated rice, favorable alleles were retained while removed undesirable ones. Most of QTLs controlling vascular bundle, rachis branches and spikelets per panicle showed cluster form or close linkage on chromosomes. The directions of their additive effects were consistent, which explained the genetic basis of significant correlation of their phenotypic characters. All the information suggested parallel evolution relationship among these traits. PMID- 15759866 TI - [Molecular mapping of a bacterial blight resistance gene Xa-25 in rice]. AB - Xa-25 was a bacterial blight resistance gene identified in a somaclonal mutant HX 3. A doubled-haploid (DH) population including 129 stable lines was derived from anther culture of a typical japanica 02428 and a typical indica HX-3 cross. The bacterial blight strain Zhe173, a typical bacterial blight strain in Yangtze River valley, was used to test the resistance or susceptible of the DH population lines, and the results showed that the resistance lines and susceptible lines were 62 and 67, respectively. A total of 300 SSR primer pairs covering 12 rice chromosomes were used for polymorphism survey of 02428 and HX-3. Among these primers, 74 showed polymorphism between the parents. Using these polymorphic SSR markers, bulked segregant analysis was conducted on the DH population. As the result, Xa-25 was located at the terminal region of the long arm of chromosome 4 between the two SSR markers RM6748 and RM1153, the map distance between Xa-25 and the two SSR markers was 9.3 cM and 3.0 cM, respectively. PMID- 15759867 TI - [Molecular mapping of two semidwarf genes in an indica rice variety Aitaiyin3 (Oryza sativa L.)]. AB - The rice dwarf variety Aitaiyin3 is derived from a semidwarf cultivar Taiyin1. Genetic analysis indicated that the dwarf phenotype in Aitaiyin3 is involved in two recessive loci. Using bulk analysis with SSR markers, the two loci were located on the chromosome 1 and chromosome 4, respectively. Moreover, the allelism test showed that the locus on the chromosome 1 is allelic to the semidwarf gene sd1, while the other one is a new locus, here was named as sdt2. Further molecular mapping showed that sd1 was linked with SSR markers in the order of RM297-RM302-RM212-OSR3-sd1 with genetic distances of 4.7 cM, 0 cM, 0.8 cM and 0 cM, respectively. The linkage relationship of sdt2 with five SSR markers was SSR332-RM1305-sdt2-RM5633-RM307-RM401 with genetic distances of 11.6 cM, 3.8 cM, 0.4 cM, 0 cM and 0.4 cM, respectively. PMID- 15759868 TI - [Development of a new type soybean germplasm with null lipoxygenase isozymes]. AB - Soybean protein is a kind of high-quality protein composed of balanced amino acids. It contains all kinds of amino acids, especially eight amino acids necessary for human, but also contains some components that are not good for human and affect food quality, such as Lipoxygenase (Lox) and Trypsin inhibitor (Ti). Nutritional value and processing quality of soybean can be improved by means of development of new variety with null Lox and Ti. In this paper, a new type soybean germplasm with null lipoxygenase isozymes was developed by Institute of Crop Sciences, Chinese Academy of Agricultural Sciences through years of biochemical marker assistant selection for null lipoxygenase by means of isoelectric focusing-polyacrylamid gel electrophoresis (IEF-PAGE) in the hybrid progenies of "96P17" (Female parent, a null lipoxygenase 2.3 line) and "93704" (Male parent, a null lipoxygenase 1.3 line). It is the first new soybean germplasm with null Lox1.2.3 genes in our country, which will contribute to soybean breeding for high quality, soybean production and utilization. In this paper, the development process of new type soybean germplasm is described. PMID- 15759869 TI - [QTL mapping of five agronomic traits in maize]. AB - Agronomic traits have significant influence on stability and adaptability in maize production. In this investigation, using a population with 266 F2:3 families from Yuyu22 (Zong3 x 87-1), two-location field tests were conducted in Wuhan and Xiangfan in 2001, with a randomized complete block design, to characterize five agronomic traits: ear height, tassel branch number, stalk diameter, days to pollen, and days to silk. Correlation analysis of field performance indicated that ear height, tassel branch number and stalk diameter were significantly positive correlative with single-plant yield, days to pollen and days to silk were highly positive correlative with each other, and tassel branch number was significantly positive correlative with stalk diameter too. Utilizing data of field tests and molecular markers, Composite Interval Mapping (CIM) method was used to localize the quantitative trait loci of these traits and 500 times permutation test was conducted to have proper LOD threshold value. As the results, total seven QTL of ear height, nine QTL of tassel branch number, eight QTL of stalk diameter, nine QTL of days to pollen, and seven QTL of days to silk were mapped on 10 chromosomes of maize; all of these QTL distributed unevenly on chromosomes and trended to cluster together. According to analysis of this investigation, the phenotype correlations of quantitative traits may result from the correlations of QTL controlling those traits. Those will be helpful to further understand genetic basis of agronomic traits in maize. PMID- 15759870 TI - An effective approach to assay plant response to environmental factors--DNA microarray. AB - With huge plant DNA data released by genome sequence project, the high-throughput DNA microarray technology has been proven an effective way to bridge the gap between plant genome sequence information and functional genomics. The technology has been used to investigate the gene expression patterns and to elucidate the role of genes. In this review, we briefly outline the features of DNA microarray and highlight its recent applications in plant biology, especially in the identification of stress-response elements and in the clarification of plant signal network. PMID- 15759872 TI - [Effects of double CO2 and O3 on growth and yields in soybean]. AB - The effects of doubled CO2 and O3 interaction on growth and yields in soybean were investigated in the open top chamber for the first time. Increasing CO2 can promote roots growth. The ability of nitrogen fixation enhanced. Plant height and base diameter increased. There are a little effect on the development stage, green leaf number and leaf area. Double O3 concentration inhibited roots and stems growth. The ability of nitrogen fixation decreased. Green leaf number and leaf area decreased and advanced senescence course. This experiment also indicated that CO2 enhancement could relax the injury and inhibition by O3 to soybean seedling. The differences of growth and the ability of nitrogen fixation in soybean between lasted double CO2 and control are less than that of single O3 concentration. Stimulation and dose-effect of increasing CO2 and O3 concentrations to soybean gradually induced more serious damage in the development late. Green leaf number and leaf area decreased. The ability of nitrogen fixation increased markedly. PMID- 15759871 TI - [Granularity distribution of airborne microbes in summer in Beijing]. AB - A study on granularity distribution of airborne microbes was conducted in details in summer, and the fluctuation regular was analyzed in Beijing. Results show that the distribution characteristics which are not changed with the different functional regions and periods are very different among airborne bacteria, fungi and actinomycete. Airborne bacteria appear skew distribution, the particles larger than 2.0 microm account for 80.0% of the total while smaller than 1.0 microm contribute 9.0% approximately. Airborne fungi are recorded with normal logarithm distribution, the particles between 1.0 microm and 6.0 microm account for 70.0% while smaller than 1.0 microm contribute 5.0% approximately. The distribution of airborne actinomycete are completely opposite to fungi, the particles larger than 8.2 microm and smaller than 1.0 microm account for 60.0% while between 3.0 microm and 6.0 microm contribute 10.0% approximately. Moreover, the granularity distribution of dominant fungi is consistent in different functional regions. But Cladosporium, Penicillium and Aspergillus are most collected in F3, F4 and F5 grades (1.0-6.0 microm) while Alternaria and nonsporing most in former four grades (>2.0 microm) of FA-I sampler, contributing 85.0%, 85.0%, 85.0%, 90.0% and 75.0% of the total respectively. The granularity of former presents normal logarithm distribution and latter skew distribution. In the past ten years, it is no change on the trend of microbial granularity distribution in Beijing, but the peak value declines from 3.0-6.0 microm to 2.0 3.0 microm. PMID- 15759873 TI - [Preliminary application of PCR-DGGE to analyzing microbial diversity in biofilters treating air loaded with ammonia]. AB - As a new DNA fingerprinting technique, denaturing gradient gel electrophoresis (DGGE) can be used to analyze the microbial diversity in different environmental samples. Three biofilters treating air loaded with ammonia will be studied with respect to microbial diversity by applying cultivation-independent molecular methods DGGE. The principal objectives of this paper include the development and application of a suitable set of molecular tools to deliver information about the possible microbial community changes within carriers during treatment of ammonia contaminated gas. Carriers were sampled at different time during operation of biofilters. After extraction and purification of the genomic DNA, the 16S rRNA genes (V3 region) were amplified by using the specific primers. These amplified DNA fragments were then separated by paralleled DGGE. The profile of DGGE showed that the same carrier at different time had the different bands' patterns. The structural diversity of the microbial community was examined by the Shannon index of general diversity H. Shannon indexes of three kinds of carriers all showed a decrease with time, which had a similar trend to NH3 removal efficiency. PMID- 15759874 TI - [Ozone concentration distribution of urban]. AB - The increase of ozone concentration in urban is one of the most important research topics on environmental science. With the increase of nitrogen oxides and hydrogen-carbon compounds which are exhausted from cars, the ozone concentration in urban is obviously increased on sunlight, and threat of photochemistry smog will be possible. Therefore, it is very important to monitor and study the ozone concentration distribution in urban. The frequency distribution, diurnal variation and monthly variation of ozone concentration were studied on the campus of Shandong University during six months monitoring. The influence of solar radiation and weather conditions on ozone concentration were discussed. The frequency of ozone concentration less than 200 microg/m3 is 96.88%. The ozone concentration has an obvious diurnal variation. The ozone concentration in the afternoon is higher than in the morning and in the evening. The maximum appears in June, when it is the strong solar radiation and high air temperature. The weather conditions also influence the ozone concentration. The ozone concentration in clear day is higher than in rainy and cloudy day. PMID- 15759875 TI - [Preparation method of Pd/Ce0.5Zr0.5O2 three-way catalysts]. AB - 1% Pd/Ce0.5Zr0.5O2(CZ) catalysts were prepared by deposition-precipitation (DP), mixing (MIX) and conventional impregnation (IMP) methods, and the effects of the preparation methods on the three-way catalytic behaviors were investigated. Hydrogen temperature-programmed reduction (H2-TPR) and in situ diffuse reflectance infrared fourier transform spectroscopy (DRIFTS) were performed to understand the Pd-support interaction and the three-way catalytic reaction mechanisms. The experiments results reveal that the catalyst prepared by deposition-precipitation (DP) method shows the strongest Pd-support interaction and the best light-off performance. In situ DRIFTS results suggest that there exists alternative route of NOx reduction over Pd-DP catalyst. And it is supposed that Pd-support interaction leads to the difference of reaction mechanism and then the diversity of light-off performances. PMID- 15759876 TI - [Factors of effects on transformation behaviors of inorganic mineral matters in desulfurizing process of burning coal]. AB - The Heshan coal and the Wansheng coal were selected to investigate the effects of Ca/S molar ratio, combustion temperature and other factors on the desulfurization efficiency when limestone as a desulfurizer. And then the possible effects of various desufurizing systems on the mineral constituents of their residues were discussed by using the X-ray diffraction analyzer. Moreover, the phase diagrams of the ternary system CaO-Al2O3-SiO2 were further used in order to find out the transformation behaviors of inorganic mineral matters in the desulfurizing processes of burning coal on the basis of the XRD results. The results show that the combustion temperature and the Ca/S molar ratio are the key factors affected the desulfurization efficiency and their residues' mineral constituents. When the temperature raised and the Ca/S ratio increased, the solid-phase reactions between CaO and some inorganic matters in coal ash will be obviously accelerated, which thus are good for the formaion of many mineral matters such as C4AF,C2AS and beta-C2S. Yet the anhydrite formed will decompose if the temperature is above 1050 degrees C and then the desulfurization efficiency will decrease. The desulfurization efficiency in different temperature is mainly decided by the existence form of the sulfur-contained mineral matters and their contents in the residue. In the experiment, the best desulfurization efficiency for the HS coal was about 90% when the Ca/S ratio was 2 or 3 and the range of the temperature was from 850 degrees C to 950 degrees C. PMID- 15759877 TI - [Impacts of the urbanization on waters non-point source pollution]. AB - Non-point source (NPS) pollution is the prominent source of water pollution in many countries, included America and China, of the world. Urban NPS pollution was attached little importance for long, compared with agriculture NPS pollution. While urbanization is the dominant form of land-use change in terms of impacts on water quality, the hydrology, other physical properties of watersheds as well as their NPS pollution potential at present. The formation of urban NPS pollution of water could be described by "source-process-sink". Urbanization has changed the source, process and sink of urban NPS pollution. A review was conducted on the international researches of urbanization impacts on NPS pollution in urban water environment from the point of view of "describe-predict and evaluation application". The studies of urbanization impacts on urban NPS pollution were focused on modeling the process of urban NPS pollution by hydrologic model, predicting the pollutants load of NPS pollution. It is a fresh methodology that the relationship between urbanization and urban NPS pollution of water was analyzed by the method of landscape change and ecological process. The research on temporal-spatial comprehensive impacts of landscape pattern changes, led by urbanization, on the urban NPS pollution will be one of the hotspots. PMID- 15759878 TI - [Flocculation and removal of water bloom cells Microcystis aeruginosa by chitosan modified clays]. AB - The kinetics of flocculation and removal of Microcystis aeruginosa by chitosan modified clays was studied. The efficiency of flocculating and removing of algal cells was greatly improved after the modification of the clays. About 80% of algae cell was removed in 0.5 hour, and 90% in 2 hours, when 11 mg/L modified sepiolite was added. Algae-removal capacities of different clays were all improved to a similar level of >90% at a total loading of 11 mg/L after being modified with chitosan. The efficiency of algae-removing was reduced when the clay loading was larger or smaller than the optimum loading. PMID- 15759879 TI - [Photoxidation reaction of Tl(I) in surface water]. AB - The valence state of thallium affecting its toxicity, distribution and mobility, photoxidation reaction of Tl(I) was studied under the radiation from high pressure arc mercury light or solar light. The results show that the low pH, the strong light intensity and UVB and VUC region are in favor of the photoxidation of Tl(I). In the case of pH = 2, only less than 1% Tl(I) remained in the solution after 10 min of irradiation, while pH = 9, with about 83% Tl(I) in the solution after 1 h of irradiation. After 5 min of irradiation, if the distance between the light source and the surface of solution is 20cm, just 4% Tl(I) remained in the solution, while the distance is 36 cm, still remained about 50%. 90% Tl(I) remained in filtered light, while less than 1% Tl(I) still remained in non filtered light. The microorganic effect is not obvious comparing with photoxidation effect in this experiment, the remained Tl(I) in excluding microorganic and microorganic experiment are all about 70%. PMID- 15759880 TI - [Isolation and activity of bacteria for the biodegradation of microcystins]. AB - The abilities of bacterial communities, which collected from the sediment and surface water of Dianchi Lake, for the biodegradation of microcystins (MCs) were firstly investigated. It was shown that the biodegradation rates of both MC-RR and LR by bacteria in sediment were apparently higher than those by bacteria on surface water. Five strains of bacteria, which have the abilities in the biodegradation of MCs, from the sediment were isolated using the liquid and solid medium containing MC-RR and LR as the carbon and nitrogen sources, which was extracted and purified from the cells of cyanobacterial bloom. Among the bacteria isolated, bacterium D was found to have a strong ability in the biodegradation of MCs. Initial MC-RR and LR of 60.1 mg x L(-1) and 38.7 mg x L(-1) were completely removed in 3 days and the average biodegradation rates per day for MC-RR and LR were 20.0 mg x L(-1) and 12.9 mg x L(-1), respectively. PMID- 15759881 TI - [Effect of nutrient on sludge settling property and bulking controls]. AB - The effect of nutrient (N and P) deficiency on activated sludge settleability and characteristics of activated sludge flocs in Sequencing Batch Reactor (SBR) fed with brewery wastewater were investigated. The results showed that the activated sludge had good settling properties at influent BOD5/N/P was 100/5/1,100/5/0.8 and 100/4/0.8 respectively. When BOD5/N/P was 100/0.94/0.31,100/2/0.4,100/3/0.6 respectively, the sludge settleability became worse due to excessive growth of filamentous. When BOD5/N/P was 100/0.94/1,100/5/0.31,100/5/0.4,100/5/0.6 respectively, non-filamentous activated sludge bulking occurred. Adding sufficient nutrient to the influent, the sludge bulking was controlled effectively and the low SVI was recovered. When increasing COD loading rate, the duration of recovery could be diminished. When nitrogen and phosphorus were insufficient synchronously, aerobic granular sludge was formed which was averagely 2mm-6mm in diameter. PMID- 15759882 TI - [Growth feature of biomass of Lemna aequinoctialis and Spirodela polyrrhiza in medium with nutrient character of wastewater]. AB - Duckweeds have an important potential in nutrient recovery from wastewater because of their rapid multiplication and high protein content in biomass. The growth rate of duckweed biomass has a direct relationship with nutrient removal and recovery. With laboratory experiments of batch culture and continuous culture, the growth curves of two duckweed species, Lemna aequinoctialis and Spirodela polyrrhiza, cultivated in different media were gotten and fitted by Logistic model. The effect of nitrogen on growth of duckweed was evaluated. Experimental results indicated that the growth curve had characteristic of sigmoidal shape and the growth rate had density-dependent characteristic. Results of statistical analysis demonstrated that Logistic model is suitable to describe the growth of single duckweed specie. The maximal growth rate from regression in medium with ammonia nitrogen was lower than those in medium with nitrate nitrogen. The maximal growth rate of Lemna aequinoctialis was higher than Spirodela polyrrhiza The paper also discussed the application of Logistic model in harvesting of duckweed biomass from wastewater. PMID- 15759883 TI - [Analysis of nitrogen, phosphorus nutrients and COD in waters of Yangtze River Estuary]. AB - The characters of the morphological, vertical and spatial distribution of nitrogen and phosphate in the water of Changjiang Estuary were discussed, according to the survey conducted in November 2003. Among the dissolved inorganic nitrogen species, nitrate accounted for 90% in the water of Changjiang Estuary. The concentration of nitrogen nutrients in the Changjiang Estuary show higher level, and 67% of sampling stations attained or exceeded the forth standard level of seawater, according to the national standard of seawater quality. Most of total phosphate in the water of Changjiang Estuary is total dissolved phosphate (TDP), which primary composition is dissolved organic phosphate (DOP). The dramatic changes of salinity in the water of Changjiang Estuary lead to the different distribution characteristics of nutrients. The concentrations of nitrate are higher inside the estuary and lower outside the estuary due to its conservatism. On the contrast, ammonium and phosphate are influenced by the change of particle concentrations and salinity, which are higher outside the estuary and lower inside the estuary. Besides, the contents of COD in the water of Changjiang estuary are very high, and the spatial distribution characteristics are higher inside the estuary and lower outside the estuary. PMID- 15759884 TI - [Polychlorinated biphenyls and organochlorine pesticides in animals from the Yangtze River Estuary and coastal areas]. AB - Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in the animals of the Yangtze estuarine and coastal areas were determined with GC-ECD. The results showed that the concentration distribution of PCBs ranged from 43.7 to 1260.4 ng/g, with an average value of 342.5 ng/g; 1.5-159.4 ng/g and 33.7 ng/g for DDTs; 0.9-77.0 ng/g and 13.0 ng/g for HCHs. The spatial distribution patterns of different contaminants in the same animal species and the same pollutant in the different animal species didn't follow obvious laws. Whereas, animals of the same species were characterized by higher contamination levels for males than females, and the small individuals than large ones. Compared with other areas at home and abroad and low-high level limits, a preliminary assessment was presented. As a result, it was shown that there existed a moderate level, to some extent, with the contamination in the animals from the study area. PMID- 15759885 TI - [Characteristics of municipal sewage chem-bioflocculation treatment process by using PCR-DGGE technology]. AB - It is reported that without cultivation, DNA could be directly extracted from environmental samples with molecular biological methods, such as polymerase chain reaction (PCR) and denaturting gradient gel electrophoresis (DGGE). To analyze the community diversity of activated sludge and bio-film in the municipal sewage, work was done to directly extrude crude DNA from activated sludge and bio-film samples, separate and amplify 16S rDNA by PCR and sequence it with DGGE. The results show the significant microbe community difference between cultivated and uncultivated activated sludge. Further research on the community diversity of two different sewage treatment processes was done and initial discussion on the microbial distribution in the same reactor and microbial structure in different experimental conditions was carried out. The sequences of several 16S rDNA DGGE fragments were determined and some possible bacteria were confirmed in comparision in GeneBank (NCBI). The results show that the PCR-DGGE technology combined with sequences determination is a feasible and efficient method for microorganism analysis in environmental sample. PMID- 15759886 TI - [Characteristic of the removal of 2,4-dichlorophenol by biological activated carbon]. AB - The adsorption characteristics and kinetics of 2,4-Dichlorophenol (2,4-DCP) by biological activated carbon (BAC) was studied through contrast experiments with conventional activated sludge alone or quartz as the carrier. The advantage and disadvantage of removing 2,4-DCP using BAC and the mechanism of this process were investigated. The results show that the method of removing 2,4-DCP by BAC is applicable in practices, and the process of BAC demonstrates high removal rate than the process of suspended activated sludge and biofilm with quartz as the carrier. In addition, the BAC process showed high resistance to shock loadings, therefore, it is suitable to be utilized at high organic loading and under long term operation. In BAC-system, activated carbon could not only adsorb 2,4-DCP but also oxidized 2,4-DCP. PMID- 15759887 TI - [Effect of growth phase on the flocculation of algal cells using clays]. AB - Four different growth phases, late lag phase, middle logarithmic phase, late logarithmic phase and early senescence phase of Microcystis aeruginosa 469 were studied for their surface hydrophobicity (xylene-water system) and surface charge (Zeta potential), followed by flocculation tests with sepiolite of 0.2 g/L. The results showed that the values of hydrophobicity were 64%, 48%, 58% and 70%, and the algal removal efficiencies after 60 min were 97%, 87%, 81% and 99% respectively, indicating a good relationship between surface hydrophobicity and removal efficiency (R = 0.7-0.8), but not the surface charge. Surface hydrophobicity and removal efficiency for the algal cells of Microcystis aeruginosa both varied with growth phase in a sequence of senescence phase > lag phase > logarithmic phase. PMID- 15759888 TI - [Activity of nitrifiers and metabolized products in a membrane bioreactor MBR under the condition of non-sludge discharge]. AB - A membrane bioreactor fed with synthetic ammonium bearing inorganic wastewater was operated under the following conditions: HRT, from 30 h to 7 h; influent NH4+ -N, 500 mg/L; non-discharge of sludge. Formation of extra-cellular polymers (ECP) and their influence on the activity of nitrifiers in the membrane bioreactor was investigated. Over the whole experimental period of 200 d, the activity of nitrifiers in terms of overall ammonium-oxidizing rate decreased from 2.1 kg/(L x d) to 1.5 kg/(L x d), and the nitrifier number using INTF absorbency measurement decreased from 0.023 x 10(-3) to 0.01 x 10(-3), although the MLSS increased from 4500 mg/L to 10500 mg/L. The ECP content increased from the initial 300 mg/L to 600 mg/L. Batch tests using the extracted ECP also indicated that the activity of nitrifiers could be inhibited by the existence of ECP. PMID- 15759889 TI - [Experiment of simultaneous denitrifying phosphorus accumulation]. AB - Denitrifying phosphorus accumulation (DNPA) and the factors affecting it were studied in a SBR with synthetic wastewater. The results showed that the sludge acclimatized under anaerobic/aerobic operation with good phosphorus removal ability, showed DNPA soon when fed nitrate instead of aeration following the anaerobic stage. Anaerobic stage was a vital premise to DNPA. If DNPA sludge was fed with nitrate prior to anaerobic stage, the DNPA would weaken even disappear. When acetate was used as sole carbon resource in the influent and nitrate did not exist in anaerobic, 1 hour of anaerobic time was optimal. NO3- -concentration in the anoxic was one of the factors affecting DNPA. When nitrate concentration was advanced from 5mg/L to 20mg/L, the percentage of DNPA increased from 11.9% to 48.7% under the condition of anaerobic(2h)-anoxic(1h)-aerobic(2h). But when the NO3- -concentration was enhanced upwards of 20mg/L, the efficiency cannot be improved. Induced DNPA did not disappear even though there was aerobic stage following anoxic stage, but the shorter the aerobic stage lasted the higher proportions of phosphorus removal via DNPA to total removal. PMID- 15759890 TI - [Dechlorination of p-dichlorobenzene by metallic catalytic reduction technology]. AB - Pd/Fe bimetallic catalysts were synthesized by chemical deposition and used to dechlorination treatment of p-DCB. Batch experiments demonstrated that the Pd/Fe bimetallic particles can effectively dechlorinate p-DCB. Dechlorination efficiency accounted for more than 90% at Pd weight ratio (Pd to Fe) 0.02% and metal to solution ratio about 4g/74mL in 90min. Dechlorination ratios were affected by reaction temperature, initial pH, Pd ratio and the addition of Pd/Fe. Chlorobenzene represented stable partially dechlorinated intermediates in the generation of benzene. PMID- 15759891 TI - [Effect of carbon and nitrogen sources on the accumulation of poly-beta hydroxybutyrate by purple non-sulfur photosynthetic bacteria]. AB - The effect of carbon and nitrogen sources on the accumulation of Poly-beta Hydroxybutyrate (PHB) by purple non-sulfur photosynthetic bacteria (PNSB)was investigated. The results showed that ammonia nitrogen and organic nitrogen could benefit the PNSB accumulating PHB; under certain ratio of carbon and nitrogen low carbon concentration can accumulate more PHB, but high carbon concentration is better for PNSB accumulating PHB with the growth of PNSB. As to different substrates, butyrate is the best for accumulating of PHB, but acetate is better and propionate is the poorest. Mixed substrate with butyrate as the main content are better than single substrate; carbon dioxide have certain effect on the PNSB accumulating PHB, and high carbon dioxide concentration can improve PHB content when using acetate or butyrate as substrate. PMID- 15759892 TI - [Regular pattern of non-point source pollutants losses in Luxi watershed]. AB - The losses of non-point pollutant combined with natural rainfall events were monitored at outlets of Luxi watershed and several field plots with different land use. The paper probes characteristics of the sediment and chemical losses during a single rainfall-runoff process. The impact of preceding precipitation is also discussed. The results of field experiments show that the concentration of SS in runoff dropped off obviously with the increase of antecedent rainfall. The observed data were 265.6, 145.8, 124.2mg/L respectively with precipitation of 15, 115, 161 mm in preceding 5 days. Compared to the losses of SS, nutrients and COD were not changed greatly by the preceding rainfalls. The fluctuations of pollutant concentration were less than 15%. The results from plots suggested that land use play an important role in pollutant losses. The following is the list ordered according to concentration of pollutant in runoff, from the highest to the lowest: Dry land,residential area, grassland, woodland and paddy land. Vegetation cover is effective on the reduction of soil erosion and sediment loss. About 80% SS could be cut down when the coverage changed from 50% to 90%. Losses of TN were mainly determined by the TN concentration in soil. The ratios of TN concentration in runoff to that in soil were 2.75%, 2.79%, 2.42 % respectively for the land use of dry land, grassland and woodland. Augment of vegetation overage contributed to increment of N losses,including NO3- -N and NH+ -N. PMID- 15759893 TI - [Continuous operation of hydrogen bio-production reactor with ethanol-type fermentation]. AB - The natural response of a continuous stirred tank reactor (CSTR) for hydrogen bio production using molasses wastewater as substrate was investigated. Emphasis was placed on assessing the operational controlling strategy on the stable operation of CSTR with high efficiency. It was found that at an initial biomass of 15g/L, an equilibrial microbial community in the ethanol-type fermentation and efficient stable operation of CSTR could be established with following conditions: temperature of 35 degrees C +/- 1 degrees C, COD organic loading rate (OLR) of 40kg/(m3 x d), hydraulic retention time (HRT) of 4h, pH value of 4.6 - 4.9 and oxidation reduction potential (ORP) of -450 - -470mV. Following that, hydrogen production in the reactor was relatively stable. The observed maximal hydrogen bio-production rate was 7.63m3/(m3 x d). The content of hydrogen in the biogas was about 40% - 58%. COD removal rate was between 22% - 26%. The total content of ethanol and acetic acid in the fermentative end products was above 80%. PMID- 15759894 TI - [Treatment performance of submerged membrane bioreactor treating brewery wastewater]. AB - Under the condition of keeping the influent COD: TN: TP = 100:5:1, submerged MBR has an excellent treatment performance for COD and NH4+ -N, and the removals for COD and NH4+ -N are both beyond 90% under steady state, in addition, MBR has a strong adaptation ability for shock organics loading rate. When the organic loading rate was increased from 0.27g/(g x d) to 0.54g/(g x d) suddenly, there was no big fluctuation for COD in effluent. According to the results of GC/MS, the remaining organics in effluent was mainly alkyl hydrocarbon, and the membrane modules played a main role in stabling permeation quality. When the sludge in MBR was at the multiplication stage, the system has a removal of about 40% for TN because of biosynthesis and simultaneous nitrification and de-nitrification, in addition, a certain removal for TP was also observed. When the sludge was at the steady stage, the removal for TN decreased to about 30% due to simultaneous nitrification and de-nitrification, whereas, the removal efficiency for TP was very little, and sometimes even below zero. PMID- 15759895 TI - [Variation of estrogenic effects during water treatment processes in a drinking water work in South China]. AB - The variation of estrogenic effects during water treatment processes in a drinking water work in South China was investigated. Water samples were collected from the source water and the different treatment processes. Crude extracts were obtained by solid phase extraction (SPE) using HLB columns and three fractionated extracts were obtained by elution using organic reagents with different polarity. Recombinant gene yeast assay was used to evaluate the endocrine disrupters effects of all the samples. The combination of the recombinant gene yeast assay and the sample fractionation technique was effective to evaluate the estrogenic effects for water samples. All the crude samples showed positive results in the yeast assay and the highest effect was occurred in the source water sample, in which the effects were mainly attributed to the none-polar fraction and the estradiol equivalent concentration was 0.25 pmol/L. 83% estrogenic effects were reduced effectively by the treatment processes. PMID- 15759896 TI - [Characteristics of granular sludge of the internal circulation anaerobic expanded bed reactor]. AB - Granular sludge characteristics of the Internal Circulation Anaerobic Expanded Bed (ICAEB) reactor is reported. Experimental results show that the mean COD removal was 89.0% and the internal circulation flow rate was about 12.5 when the reactor running steadily with the mean COD volumetric loading rate 28.2kg/(m3 x d). General granular sludge and suspended granular sludge are cultured in the reactor. The suspended granular sludge layer found in the fluid level of the second reaction section is dynamically stable. Keeping comparative steady operational condition could reduce the adverse influence of the suspended granular sludge layer, if necessary, suitable measurement could be adopted to remove the overabundant suspended granular sludge off. PMID- 15759897 TI - [Influence of humic substance in solids on CODMn of the Yellow River]. AB - Experiments were carried out to study the influence of humic substance in solids (Loess soil) on CODMn in the Yellow River. Several significant results were abtained from the study: (1) The total CODMn of water sample including liquid and solid phases increased significantly with the increase of solid content; The CODMn of liquid phase of water sample under pre-treatment with acid (LPWSPC) also increased significantly with the increase of solid content while that without pre treatment did not increased significantly. (2) With the increasing of solid content, the total BOD5 of water sample increased much slower than the total CODMn and the CODMn of LPWSPC. (3) The humic substances with the content of 0.76% in solids (Loess soil) contributed greatly to the total CODMn of water sample. When the solid contents were 7.5 g/L and 15.0 g/L, the humic substances in solids accounted for 15.9% and 21.7% of the total CODMn of water sample, respectively. (4) When solid contents were 7.5 g/L and 15.0 g/L, fulvic acid,one of the main compositions of humic substance contributed to 23.6% and 50.6% of the CODMn of LPWSPC, respectively. Since the fulvic acid can hardly be biologically oxidized under natural conditions, the CODMn of LPWSPC in water quality monitoring exaggerates the oxygen-consuming organic contamination of the Yellow River. PMID- 15759898 TI - [Streaming current of waterworks sludge]. AB - The applicability of streaming current in waterworks sludge treatment as indicator of coagulant dosage and pH control was investigated in this study. Experiments were conducted on sludge settling behaviors and dewatering processes. At the aspect of sludge settleability research showed the theoretical optimal dosage, pH was 8.9mg/L, 7.25, and the practical optimal dosage, pH was 9.0mg/L, 6.90. On the side of sludge dewaterability, the theoretical optimal dosage, pH was 25.0mg/L, 9.25, and the practical optimal dosage, pH was 26.0mg/L, 9.0. Results indicated that slightly positive streaming current coincided with the optimal dosage and pH. Thus streaming current can be used for continuous dosage feedback control as the single indicator. PMID- 15759899 TI - [Three-dimensional excitation emission matrix fluorescence spectroscopic characterization of the complexation between mercury (II) and dissolved organic matter]. AB - With the development of fluorescence technique, three-dimensional excitation emission matrix fluorescence spectroscopy (3DEEM) was widely applied to characterize the nature of dissolved organic matter (DOM) in natural waters since the last decade. 3DEEM and fluorescence quenching titration were used to study the interaction between Hg(II) and DOM. The results show that a general decrease in intensity for individual peak A, B and C was found as concentrations of Hg(II) increased. pH is the main parameter that strongly influences the Hg(II)-DOM complexation. The addition of Cl- ion to the Hg(II)-DOM system caused an increase in fluorescence intensity. The results also show that the addition of Ca(II) solution strongly enhanced the fluorescence intensity for humic-like fluorescence, while the protein-like fluorescence only slightly enhanced. On the contrary, no fluorescence enhancement was found in the Hg(II)-DOM system after the addition of Mg(II) ion. A decrease in fluorescence emission intensity was found after the addition of Cu(II) ion to the Hg(II)-DOM solution. PMID- 15759900 TI - [Sorption and desorption of hydrophobic organic contaminants (HOCs) by different type of coal]. AB - To understand the intercorrelation between the heterogeneity of kerogen and sorptive-desorptive characteristics, the sorption and desorption equilibria of phenanthrene and 1,3,5-trichlorobenzene were measured for three different types of coal (lignite, bark coal and fusinite) using a batch technique. The results show that the sorption and desorption of phenanthrene and 1,3,5-trichlorobenzene are highly nonlinear and significantly hysteretic. The observed sorptive differences among the three coals are explained viathe structural and chemical heterogeneity of kerogen. PMID- 15759901 TI - [Stratification of free air space and its dynamics in the process of co composting]. AB - Co-composting of sewage sludge and pig manure was studied. Free air space (FAS) in the whole composting pile was 26.2% at the temperature rising stage and 42.3% at the stage of high temperature composting. FAS in the upper portion of composting pile was higher than that in the lower, where the air was enough for microorganism to live on for two stages above. The FAS was 52.4% at the stage of the cooling, when the effect of stratification of FAS was significant. The FAS in the upper pile was bigger than that in the middle, which was bigger than that in the lower. At the stageof maturing, FAS was 54.7% and its stratified effect weakened, but the capability of aeration in the upper and in the middle of the pile was larger than that in the lower. At the different stages of composting, the degree of difference of FAS was of the order: the cooling stage > the high temperature stage > maturing stage > the temperature rising stage. The difference in the pile for every stage of composting was as follows: the upper > the middle > the lower. The change of FAS agreed with two-order kinetics. PMID- 15759902 TI - [Carbon output through urban domestic garbage in Haikou]. AB - Carbon output through urban domestic garbage is one part of carbon cycle in urban ecosystem which is related to some environmental pollution. Researches on carbon output through urban domestic garbage is the foundational work on carbon cycle in urban ecosystem, but also can help to analyze the influences of residents' consumption on environment. Based on data got through sampling and collecting, the status and trend of carbon output through domestic garbage in Haikou was analyzed. The results showed that, the total carbon output through 26.92 x 104 t of domestic garbage is 10.51 x 104 t and the carbon output per capita everyday is 0.326 kg/(cap. d)(-1) in 2001. About 31% of the total carbon will be transferred into methane, carbon dioxide and organic matter in extravasate. About 69% of the total carbon will be sequestrated in landfill. Because 33% of the sequestrated carbon originate from atmosphere, urban domestic garbage may be a carbon sink. From 1991 to 1999, the carbon output through Haikou's domestic garbage is annually increasing by 9.55%, and the output of sequestrated carbon is annually increasing remarkably by 13.23%. The increase of carbon output is significantly correlated with the increase of expenditure per capita and population, which shows expenditure per capita and population have some effect on carbon output. PMID- 15759903 TI - [Configuration of pyrolytic chars from waste tires in fluidized bed reactor]. AB - With the fluidized bed as main reactor, the configuration of chars of waste tire was investigated. The change of specific surface area, porosity and specific pore volume of chars received at various temperature, diameter of bed materials and superficial fluidization number was mainly researched. The specific surface area and porosity of chars had the peak value at 650 degrees C or 750 degrees C, which showed there exists the best pyrolysis temperature from the angle of char quality and it will decrease with smaller diameter of bed materials, 0.135-0.304mm. The porosity of chars decreases with the fluidization number increasing. The change tendency of the specific surface area of chars with the fluidization number is correlated with the pyrolysis temperature. At 550 degrees C the specific surface area of chars decreases with the fluidization number increasing, while at 650 degrees C the other way round. The change tendency of the specific pore volume of chars with the temperature is correlated with the diameter of bed materials. With larger bed materials (0.304-0.4mm), the specific pore volume of chars rises at fisrt, then decreases with the temperature increasing, and with smaller bed materials, it decreases with the temperature increasing. PMID- 15759904 TI - [Problems and solutions of implementing plan environmental impact assessment in China]. AB - At present, there are two forms of Plan Environmental Impact Assessment (PEIA): Self-assessment and entrusted assessment. The appropriate object and the time of starting PEIA were discussed. It points out that self-assessment should be able to realize the essential roles of PEIA, such as 'starting as early as possible', 'optimization of options', etc. At the same time, the concept and function of alternative, PEIA methodology and the role of public participation are all elementarily discussed. PMID- 15759905 TI - Use of computerized clinical support systems in medical settings: United States, 2001-03. AB - OBJECTIVES: This report presents information on the use of electronic clinical systems to support patient care in physician offices and hospital emergency and outpatient settings. Percentages of hospital emergency and outpatient departments with electronic patient medical records and automated drug dispensing systems are presented by selected hospital characteristics for 2001-02. Percentages of physicians using electronic patient billing records, electronic patient medical records, and computerized prescription order entry systems are presented by selected physician characteristics for 2003. METHODS: Data are from provider induction interviews from the National Ambulatory Medical Care Survey (NAMCS) and the National Hospital Ambulatory Medical Care Survey (NHAMCS), the ambulatory care component of the National Health Care Survey (NHCS). The NHCS is a family of provider-based surveys that collect information on the care provided in various medical care settings. RESULTS: During 2001-03, electronic medical records were used less often in physician offices (17 percent) than in hospital emergency (31 percent) and outpatient departments (29 percent). In physician offices, information technology was more frequently used for billing patients (73 percent) than for maintaining medical records electronically (17 percent) or ordering prescriptions electronically (8 percent). Automated drug dispensing systems were available in hospital emergency departments (40 percent) more frequently than in outpatient departments (18 percent). PMID- 15759907 TI - [Traffic accidents: increasingly frequent and fatal]. PMID- 15759906 TI - [Role of the family in the suicidal attempt of the adolescent]. PMID- 15759908 TI - [Social health research: recent communications in Salud Publica de Mexico]. PMID- 15759909 TI - [Prevalence of violent events and post-traumatic stress disorder in the Mexican population]. AB - OBJECTIVE: To report the rate of exposure to different violent events, their demographic correlates, the prevalence of Post-Traumatic Stress Disorder (PTSD), and the impact on quality of life. MATERIAL AND METHODS: The National Survey of Psychiatric Epidemiology is representative of the Mexican urban population aged 18 to 65. The survey was undertaken in 2001 and 2002 using the Composite International Diagnostic Interview (CIDI- 15) computerized version. The statistical analyses take into account the multistage, stratified, and weighted sample design. Kaplan-Meier and logistic regressions were performed. RESULTS: Sixty-eight percent of the population has been exposed to at least one stressful life event. Exposure varies by sex (rape, harassment, and sexual abuse are more frequent in women; accidents and being a victim of burglary among men) and by age (more frequent in children, adolescents, young adult women, and the elderly). By sex, 2.3% of women and 0.49% of men present PTSD. Rape, harassment, kidnapping, and sexual abuse are the events most associated with PTSD. CONCLUSIONS: The results suggest the need to increase treatment coverage to attend the consequences of violence, taking into consideration the important gender and age variations. PMID- 15759910 TI - [Psychopathology across three family generations: an epidemiological study in Mexico City]. AB - OBJECTIVE: To estimate the risk of psychiatric disorders across three family generations. MATERIAL AND METHODS: The research design corresponds to a "family history study" and is based upon a household survey conducted in 1995, on a representative sample of Mexico City's adult population aged 18 - 65 years (n=1932). Briefly, a standardized assessment of adults' lifetime prevalence of psychiatric disorders was obtained using an amended version of the Composite International Diagnostic Interview, CIDI. In addition, all respondents provided information on their parents' psychiatric history of anxiety, affective disorders, and substance-use disorders.Also, respondents who had 4-16 year old children living in the same household were interviewed using a standardized questionnaire that included specific questions on their children's psychiatric symptoms (n=925 parents, corresponding to 1686 children and adolescents). Familial psychopathology across generations was defined as follows: a) history of psychiatric disorders only in grandparents; b) history of psychiatric disorders in one parent only (mother or father); and, c) history of psychiatric disorders history in grandparents and in parent. The risk of psychiatric morbidity was estimated via odds ratios obtained from logistic regression models with and without random-effects and using Generalized Estimation Equations with robust variance estimation. RESULTS: The risk for psychopathology in the offspring across generations was estimated to be two to three times higher when psychiatric disorders occurred among the parents. The risk of psychiatric disorders was estimated to be higher when comorbidity among different types of disorders was present. Conclusions. The results suggest that the risk for developing psychopathology across generations is at least moderate and relatively stable. Familial transmission of psychopathology across three generations seems to resemble a genetic mixed model. PMID- 15759911 TI - [Factors related with the severity of injuries in public streets of an urban area in Cuernavaca, Morelos,Mexico]. AB - OBJECTIVE: To identify risk factors associated with severity of injuries occurring in public streets of an urban area. MATERIAL AND METHODS: Between February and April 2001, a cross-sectional survey was conducted among injured people seeking emergency care at three hospitals in Cuernavaca, Morelos, Mexico. Information was also obtained for those who died on accident sites. Data on the following variables were collected: age, sex, alcohol intake, day and time of injury, prehospital care, external cause and nature of injury. The Abbreviated Injury Scale (AIS) was used to define severity of injuries. Data analysis consisted of descriptive, bivariate, and multivariate regression models. RESULTS: A total of 492 injured people were included in the study; 23 of them died on site. The main cause of injuries was road traffic accidents (52%); 72% of injuries were not severe. Variables associated with the severity of injuries were: Road traffic accidents (odds ratio [OR] 6.59, 95% confidence interval [CI] 2.52, 17.23), adjusted for age and schooling. CONCLUSION: Road traffic injuries are the main cause of severe injuries. TheAIS is useful to assess the frequency and distribution of injury severity. PMID- 15759912 TI - [Screening and follow-up for cervical cancer prevention in rural Mexico using visual inspection]. AB - OBJECTIVE: To compare the standard cervical cancer screening procedure--the Papanicolaou test or Pap smear--with detection through visual inspection using acetic acid (VIA), and visual inspection with acetic acid assisted by Aviscope (VIAM). MATERIAL AND METHODS: The study was conducted between October 1998 and December 2000, in two Mixteca regions in Oaxaca, Mexico. A field trial design was used to randomize the two regions to either of the two inspection methods. In one region 2,240 women received VIA; in the other, 2,542 women received VIAM. Women positive to visual inspection and a subsample of women with negative results were referred for colposcopy and, if necessary, cervical biopsy. Statistical data analysis consisted of descriptive statistics and comparison of groups using chi squared tests. Diagnostic values were obtained for VIA and VIAM, using as gold standards colposcopy and biopsy. RESULTS: VIAM identified a higher proportion of women with a cervical abnormality (16.3%) than VIA (3.4%), as well as normal women (58.5% vs. 53.8%).VIA identified a higher proportion of women with benign changes (41.2%) than VIAM (19.6%).VIAM had a greater sensitivity (p>0.05) but lower specificity (p<0.05) than VIA, using as gold standards colposcopy and biopsy. CONCLUSIONS: The two study groups were comparable for age, parity and cytological results; however, visual inspection results were different between the groups: VIAM identified more lesions confirmed by biopsy. The diagnostic utility of VIAM and VIA was lower than expected. Training of clinical personnel in visual inspection is critical to improve the effectiveness of these screening methods. PMID- 15759913 TI - [Breast feeding and the incidence of acute diarrhea during the first three months of life]. AB - OBJECTIVE: To assess the effect of exclusive breastfeeding and its interactions with selected socioeconomic factors, on the incidence of acute infantile diarrhea during the first three months of life. MATERIAL AND METHODS: A prospective cohort study with a follow up of three months was conducted in 327 mother and child pairs, living in the city of Durango, Mexico, between April and June 1994. Data analyses included simple univariate and bivariate cross-tabulations, as well as multivariate logistic regression models with random effects. RESULTS: Infants who were not exclusively breastfed were at a significantly increased risk of having acute diarrhea during early infancy (ORcombined breastfeeding=3.23; 95% CI 1.84 5.68 and ORartificial breastfeeding=4.36; 95% CI 2.32-8.19). Moreover, the protective effect of breastfeeding was independent from the effects of the following factors: poor maternal education, lack of social support for baby care, and being an adolescent mother. However, lack of potable water and lack of sewerage were potential effect modifiers of type of lactation deltaG=9.26; p=0.09; ORno water/no sewerage= 2.58; 95% CI 1.10-6.03 in the final multivariate model), for simultaneous lack of potable water and sewerage, which is greater than the sum of the individual OR for each variable. CONCLUSIONS: Similar to previous studies, exclusive breastfeeding was found to have great importance, since this practice protects the child's health and allows for a better development, despite unfavorable social and economic conditions. In particular, it seems that the protection conferred by breastfeeding is stronger when home conditions are poor, but this same condition presents an intervention opportunity, particularly the assurance of potable water provision for the infant when breastfeeding is not an option. PMID- 15759914 TI - [Estimation of exposure to fluoride in "Los Altos de Jalisco", Mexico]. AB - OBJECTIVE: To estimate the level of fluoride exposure and human health risks in Los Altos de Jalisco (Jalisco State Heights) region. MATERIAL AND METHODS: This study was conducted between May and July 2002. The fluoride concentrations of 105 water wells and six tap water samples were electrochemically measured. Exposure doses to fluoride and total intake of fluoride were estimated for babies (10 kg), children (20 kg), and adults (70 kg). RESULTS: The fluoride concentration of the water samples ranged from 0.1 to 17.7 mg/l. More than 45% of the water samples exceeded the national guideline value for fluoride of 1.5 mg/l. The estimated values of the exposure doses to fluoride and total intake of fluoride were in the range of 0.04-1.8 mg/kg/d and 0.5-18.4 mg/d, respectively. CONCLUSIONS: Dental fluorosis, skeletal fluorosis, and bone fractures are some of the potential health risks due to the intake of high doses of fluoride for the population of Los Altos de Jalisco. In order to reduce health risks, fluoridated salt,fluoridated toothpastes, and drinking water containing more than 0.7 mg/l of fluoride should be avoided. PMID- 15759915 TI - [Prehospital emergency care in Mexico City: the opportunities of the healthcare system]. AB - Unintentional vehicle traffic injuries cause 1.2 million preventable deaths per year worldwide, mostly affecting the population in their productive years of life. In Mexico, unintentional vehicle traffic injuries are one of the main causes of death; in Mexico City they account for 8% of deaths. Prehospital systems are set up to provide hospital medical care to the population, by means of a complex network that includes transportation, communications, resources (material, financial and human), and public participation. These systems may be designed in a variety of ways, depending on availability, capacity and quality of resources, according to specific community needs, always abiding by laws and regulations. In Mexico, several institutions and organizations offer prehospital services without being overseen in terms of coordination, regulation and performance evaluation, despite the high rates of morbidity and mortality due to injuries and preventable conditions amenable to effective therapy during the prehospital period. Prehospital care may contribute to decrease the morbidity and mortality rates of injuries requiring prompt medical care. Emphasis is made on the importance of assessing the performance of prehospital care, as well as on identification of needs for future development. PMID- 15759916 TI - [One foot sleeping. Facing health systems]. PMID- 15759917 TI - [An interview of Salud Publica de Mexico with Dr. Jaime Mier y Teran, Ministry of Health of Tabasco]. PMID- 15759918 TI - [An interview of Salud Publica de Mexico with Dra. Asa Cristina Laurell, Ministry of Health of Distrito Federal]. PMID- 15759919 TI - [An interview of Salud Publica de Mexico with doctor Alfonso Petersen Farah, Ministry of Health of Jalisco]. PMID- 15759920 TI - [Experimental study on repair of goat tibia defect with marrow stromal cell and bio-derived bone]. AB - OBJECTIVE: To investigate the feasibility of repairing goat tibia defect with marrow stromal cells (MSCs). METHODS: MSCs were cocultured with the bio-derived bone in vitro, and the 20 mm tibia defects were made and fixed with plate in 35 goats, and they were divided into the experimental group, control group and blank group. The defects on the right side were filled with tissue engineering bone as the experimental group, the defects on the left side with bio-derived bone as the control group in 33 goats, and the defect on the both sides were not filled with any materials as the blank group in 2 goats. The repair capability was assessed physically, histopathologically and biomechanically at 2, 4, 6, 8, 12, 16 and 24 weeks after operation in 3 groups. RESULTS: By physical, histopathological and biomechanical examinations, the bio-derived bone was partially absorbed in the experimental group and was rarely absorbed in the control group in the 4th week; the defects were partially repaired in the experimental group, and in the control group, few new bones were observed in the two ends of the implants, in which there was fibrous tissue. The effects of biomechanics had no statistically significant difference between the experimental group and the control group (P>0.05) in the 8th week; the defects were perfectly repaired in the experimental group and the effects of biomechanics had statistically significant difference between two groups (P<0.05) in the 12th weeks. The defects were not repaired in the 24th week in the blank group. CONCLUSION: The tissue engineering bone can efficiently repair bone defect, and its repair capability is better than that of bio-derived bone alone both in quantity and in quality of bone formation. PMID- 15759921 TI - [Experimental study on repairing segmental bone defect with bio-derived bone preserved by various methods]. AB - OBJECTIVE: To study the difference of repairing segmental bone defect with bio derived bone preserved by various methods. METHODS: Freeze-dried biomaterials had been stored in two different preservation solutions for three months, while the biomaterials stored for same period were observed as control group. The experimental model of 15 mm radial segmental defect was made in 60 New Zealand white rabbits, which were divided into groups A, B and C according to transplant materials preserved by various methods. Groups A and B were deeply divided into A1 and A2 subgroups, B1 and B2 subgroups according to whether materials were cocultured with osteoblasts. Tissue engineered bone was used to repair bone defects of left limbs in A1 and B1 subgroups, while simple material to repair defects of right limbs in A2 and B2 subgroups. Group C was divided into C1 and C2 subgroups. Freeze-dried material was used to repair bone defects of the left limbs, while defects of the right limbs as blank control group. The samples were harvested and observed by the roentgenographical, histomorphological, biomechanical and computerized graphical analysis at 4, 8 and 16 weeks. RESULTS: All of the defects treated with implants exhibited new bone formation 4, 8 and 16 weeks postoperatively, increasing with time. The radiological, histomorphological and biomechanical evaluation showed that the ability of new bone formation was arranged in 6 subgroups as follows: A1>A2>C1>B1>B2>C2, the difference was significant between them (P<0.001, P<0.05). The ability of new bone formation was strongest and at 16 weeks the defect was bridged with the appearance of marrow cavities in A1 subgroup, the biomechanical properties in implants approached to those of normal bone. CONCLUSION: The choice of proper preservation solution can improve the ability of repairing bone defect. PMID- 15759922 TI - [Tissue engineered bone regeneration of periosteal cells using recombinant human bone morphogenetic protein 2 induce]. AB - OBJECTIVE: To investigate bone regeneration of the cell-biomaterial complex using strategies of tissue engineering based on cells. METHODS: Hydroxyapatite/collagen (HAC) sandwich composite was produced to mimic the natural extracellular matrix of bone, with type I collagen serving as a template for apatite formation. A three-dimensional poly-porous scaffold was developed by mixing HAC with poly(L lactic acid) (PLA) using a thermally induced phase separation technique (TIPS). The rabbit periosteal cells were treated with 500 ng/ml of recombinant human bone morphogenetic protein 2 (rhBMP-2), followed by seeded into pre-wet HAC-PLA scaffolds. Eighteen 3-month nude mice were implanted subcutaneously cell suspension (group A, n = 6), simple HAC-PLA scaffold (group B, n = 6) and cell biomaterial complex (group C, n = 6) respectively. RESULTS: Using type I collagen to template mineralization of calcium and phosphate in solution, we get HAC sandwich composite, mimicking the natural bone both in composition and microstructure. The three dimensional HAC-PLA scaffold synthesized by TIPS had high porosity up to 90%, with pore size ranging from 50 microm to 300 microm. SEM examination proved that the scaffold supported the adhesion and proliferation of the periosteal cells. Histology results showed new bone formation 8 weeks after implantation in group C. The surface of group A was smooth without neoplasma. Fibrous tissue invasion occurred in group B and no bone and cartilage formations were observed. CONCLUSION: The constructed tissue engineering bone has emerged as another promising alternative for bone repair. PMID- 15759924 TI - [Effects of platelet-rich plasma on proliferation and osteogenetic activity of marrow mesenchymal stem cells in vitro]. AB - OBJECTIVE: To study the effects of platelet-rich plasma (PRP) on the proliferation and osteogenetic activity of the marrow mesenchymal stem cells (MSCs) cultured in vitro to elucidate the cellular and molecular mechanism by which PRP accelerates bone repair. METHODS: The human MSCs were cultured in vitro and randomly divided into the experimental group (n = 9) and control group (n = 9). In the experimental group, the MSCs were inter-fused with PRP (10 microl/ml culture media). The proliferation ability of the cells was tested by flow cytometry and MTT, the osteogenetic activity by alkaline phosphatase (ALP) measuring and tetracycline fluorometry, and cbfa1 mRNA expression by reverse transcript-PCR. RESULTS: PRP could stimulate the MSCs proliferation. The flow cytometry assay showed that the MSCs proportion of S period of the experimental group significantly increased 14.5+/-0.4 in comparison with that of the control group 7.2+/-0.5 (P<0.01) after 24 hours. MTT value showed that MSCs proliferated to platform period earlier in the experimental group than in the control group. There was a significant increase in ALP activity of the experimental group 7.79+/ 1.98, 9.51+/-2.31 and 14.03+/-3.02 when compared with that of the control group 2.06+/-0.77, 2.84+/-0.82 and 2.58+/-0.84 after 3, 6 and 9 days (P<0.05). The number of mineral nodes increased. Reverse transcript-PCR showed that the expression of cbfa1 mRNA were elevated gradually at 2, 4 and 8 hours after inter fused with PRP. CONCLUSION: The effect of PRP on accelerating bone repair is related to its effects on stimulating the proliferation of MSCs, increasing the cbfa1 expression and promoting the osteogenetic activity. PMID- 15759923 TI - [Expression of interleukin 2 and IL-2 receptor after implanted tissue engineered bones constructed with allogeneic marrow stromal stem cells and bio-derived materials in rhesus monkeys]. AB - OBJECTIVE: To explore the feasibility of allogeneic marrow stromal stem cells (MSCs) as seed cells to construct tissue engineered bone by detecting the expressions of interleukin 2 (IL-2) and IL-2 receptor in rhesus monkeys after implanting these tissue engineered bones. METHODS: Engineered bones were constructed with osteoblasts which derived from allogeneic MSCs and bio-derived materials in vitro, and then were implanted to bridge 2.5 cm segmental bone defects of left radius in 15 rhesus monkeys as experimental group, bio-derived materials only were implanted to bridge same size defects of right radius as control group. Every 3 monkeys were sacrificed in the 1st, the 2nd, the 3rd, the 6th and the 12th weeks postoperatively and the expressions of IL-2 and IL-2 receptor in blood and graft samples were detected quantitatively by enzyme-linked immunosorbent assay (ELISA). RESULTS: There was no significant difference in the contents of IL-2 and its receptor between 2 groups (P>0.05). The contents of IL-2 and its receptor increased from the 2nd week and maintained high level from the 2nd to the 6th week, but decreased after 6 weeks. CONCLUSION: Tissue engineered bones constructed with allogeneic MSCs and bio-derived materials show low immunogenicity. Allogeneic MSCs may be used as seed cells to construct tissue engineered bone. PMID- 15759925 TI - [In vivo ectopic osteogenesis of nacre/polylactic acid artificial bone combined with allogenic osteoblasts]. AB - OBJECTIVE: To study the mechanism of ectopic osteogenesis of nacre/Polylactic acid (N/P) artificial bone combined with allogenic osteoblasts, and to explore the possibility as a scaffold material of bone tissue engineering. METHODS: The allogenic osteoblasts seeded onto N/P artificial bone were co-cultured in vivo 1 week. The N/P artificial bone with allogenic osteoblasts were implanted subcutaneously into the left back sites of the New Zealand white rabbits in the experimental group and the simple N/P artificial bone into the right ones in the control group. The complexes were harvested and examined by gross observation, histologic analysis and immunohistochemical investigation 2, 4 and 8 weeks after implantation respectively. RESULTS: In experimental group, the osteoid formed after 4 weeks, and the mature bone tissue with bone medullary cavities formed after 8 weeks; but in control group there was no new bone formation instead of abundant fibrous tissue after 4 weeks, and more fibrous tissue after 8 weeks. CONCLUSION: N/P artificial bone can be used as an optical scaffold material of bone tissue engineering. PMID- 15759926 TI - [Experimental study on repairing segmental bone defects with three bio-bone derived materials]. AB - OBJECTIVE: To evaluate the osteogenesis of three bio-bone derived materials in repairing segmental bone defects. METHODS: Sixty Japanese rabbits were made 10 mm radius segmental defects and divided into 5 groups (groups A, B, C, D and E, n=12). Composite fully deproteinised bone (CFDB, group A), partially deproteinised bone (PDPB, group B), partially decalcified bone (PDCB, group C), autogenous iliac bone graft (group D) and no implant (group E) were implanted into the radius segmental bone defects of rabbits. The specimens were examined after 4, 8, 12 and 24 weeks; the osteogenesis was evaluated through X-ray radiograph and undecalcified solid tissue histological examination. RESULTS: The border between the material and host's bone was distinct after 4 weeks and blurred after 8 weeks; the density of partial edge of the material was similar to that of radii after 12 weeks. The medullary cavity of bone reopened in group B; the density of most defect area was similar to that of the host bone and there was a few high density shadow in group C; the density of most defect area was higher than that of host bone in group A after 24 weeks. There was no significant difference in radiograph scoring between groups A, B and C after 4 weeks and 8 weeks (P>0.05); the scores of group B and C were higher than that of group A after 12 weeks (P<0.05); and the scores were arranged as follow: group D > group B > group C > group A after 24 weeks (P<0.05). Bone callus grew toward defect area and new bone adhered to the material after 4 weeks and 8 weeks; more new bone formed, and the materials were absorbed and degraded with time. The quantity of bone formation was more in group D than in group B and in group B than in group C and in group C than in group A after 24 weeks (P<0.05). CONCLUSION: PDPB had good osteogenesis in repairing the segmental bone defect, PDCB was inferior to it, both PDPB and PDCB are fit to repair segmental bone defect. Both of them were inferior to autogenous bone. PMID- 15759927 TI - [Study on transplantation of marrow stromal stem cells into acellular extra cellular matrix in mice [corrected]]. AB - OBJECTIVE: To study the integration of mice [corrected] marrow stromal stem cells (MSCs) after transplantation into acellular extracellular matrix (AECM). METHODS: We got 16 femurs from 8 Kunming mice [corrected], the femurs were treated by Triton X-100 to get AECM, MSCs were collected from femoral marrow of 20 Kunming rats about a mouth old by PBS 4 ml, centrifuged and primary cultured in bottles, then the mice [corrected] MSCs were transplanted into AECM at a concentration of 5 x 10(6)/ml and cultured for 7 days. The integration of the donor cells was observed using one phase contrast microscope, a light microscope and a scanning electron microscope (SEM). RESULTS: In AECM bone lacunas there were MSCs nuclei stained blue. The nuclei were unevenly distributed in AECM with more in the peripheral AECM than in the central AECM and with more in the layer near culture medium than in the layer far away from culture medium. AECM possessed a good spatial scaffold structure, the marrow stromal stem cells were well integrated into AECM. CONCLUSION: AECM can be used as a good scaffold material for tissue engineered bone construction. PMID- 15759928 TI - [Different expression of chaperone interacting protein in normal, scar and chronic ulcer tissues and its relationship with wound healing]. AB - OBJECTIVE: To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. METHODS: Twenty biopsies including scar tissues (n = 8), chronic ulcer tissues (n = 4) and normal tissues (n = 8) were used in this study. The immunohistochemical staining (power vision two-step histo-staining reagent) was used to explore the amount and expression characteristics of such protein. RESULTS: The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen in inflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. CONCLUSION: Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing. PMID- 15759929 TI - [Repair of face soft tissue defect with prefabricated neck expander flap with the vessels of temporalis superficialis]. AB - OBJECTIVE: To probe the principle and the method to repair facial soft tissue defect with the prefabricated expander flap the neck with the vessels of temporalis superficialis. METHODS: The expander was implanted into the surface layer of the platysma in neck. The pedicle of the expander flap contained the arteria temporalis superficialis and its ramux parietalis. After 3 months, the prefabricated island expander flaps pedicled with the arteria temporalis superficialis and its ramux parietalis could be transferred to the face. From 1998 to 2003, 6 cases of facial soft tissue defects were repaired. The maximal flap size was 12 cm x 8 cm. The pedicle length was 7-8 cm. RESULTS: After a follow-up of 3-6 months, all expander flaps survived. The excellent function and cosmetic result were achieved. CONCLUSION: The prefabricated expander flaps of the neck pedicled with the arteria temporalis superficialis and its ramux parietalis can be transferred to the upper face to repair tissues defect. The supply of blood of the prefabricated expander flaps were safe and reliable. The survived areas of the flaps are directly proportional to the areas of temporalis superficialis fascia combining the expander flaps. PMID- 15759930 TI - [Reform of the pedicled abdominal flap and its clinical application]. AB - OBJECTIVE: To investigate the closing method of wound after removal of the traditional pedicled abdominal flap. METHODS: According to the design, the pedicled abdominal flaps were cut and lifted, and then the incision were extended from both sides on base of the flap to anterior superior iliac spine, respectively. After separating on superficial fascia, two flaps were obtained. The wound of donor site was closed completely by these two pedicled flaps. Twelve patients with skin defects on hands or forearms were treated using the reformed method of traditional pedicled abdominal flap. RESULTS: All of the 12 reformed pedicled abdominal flaps survived, and only one had local necrosis on the distal part of the abdominal flap, about 1.5 cm x 2.0 cm. CONCLUSION: This new design could provide a good method to close the abdominal wound after removal of pedicled abdominal flap. PMID- 15759931 TI - [Repair of perineal and adjacent defects with thoracic biblical island flaps]. AB - OBJECTIVE: To report the experience of repairing perineal and adjacent defects with thoracic biblical island flaps. METHODS: From January 1988 to October 2003, 7 cases of perineal and adjacent soft tissue defects with thoracic biblical island flaps, aged 17-52 years. Of 7 cases, there were 2 cases of severe scar contracture due to burn on perineal, 1 case of malignancy on perineal, 4 cases of vast soft tissue defects of trauma on the parts of groin and higher two-third thigh. The area of flaps was 9 cm x 27 cm-12 cm x 30 cm, the longest pedicle of blood vessel was 16 cm. The donor sites of flaps less than 10 cm could be sutured directly, the ones more than 10 cm could be repaired with skin grafting. RESULTS: All the flaps primarily survived. There was no ischemia and necrosis at the distal part of flaps. Four patients were followed up 6 months to 6 years. The color, texture and appearance of the flaps were good. The functions of walk and squat were satisfactory. CONCLUSION: The thoracic biblical island flap can repair perineal and adjacent soft tissue defects, moreover the donor is hidden and the effect is ideal. PMID- 15759932 TI - [Application of epiglottic in reconstruction of traumatic laryngotracheal stenosis]. AB - OBJECTIVE: To investigate the application and long-term results of epiglottic in reconstruction of the traumatic laryngotracheal stenosis. METHODS: From January 1988 to February 2002, 42 patients with traumatic laryngotracheal stenosis were treated, including 33 laryngeal stenosis and 9 laryngotracheal stenosis. The following surgical treatment were performed: (1) lowered epiglottic and bi pedicled sternohyoid myofascial flap and (2) lowered epiglottic and bi-pedicled sternohyoid myofascial flap and sternocleidomastoideus clavicle membrane flap. RESULTS: Thirty-seven patients (88.1%) were successfully extubated 10 to 75 days after operation. Feeding tube lasted from 9 to 24 days, all the patients rehabilitated deglutition after surgery. The time of using stent was 9 to 19 days in 25 cases. All patients were followed up 1 year to 3 years and 4 months. The function of larynx recovered completely in 37 extubated patients and partially in 5 cannulated patients. CONCLUSION: Epiglottic has the advantages of easy gain, high anti-infection and survival rate, and stable structure. A combination of epiglottic and the bi-pedicled sternohyoid myofascial flap plus sternocleidomastoideus clavicle membrane flap can repair large laryngeal and tracheal defects. PMID- 15759933 TI - [Biomechanical evaluation of the valgus stability of elbow after reconstruction]. AB - OBJECTIVE: To evaluate of the valgus stability of the elbow after excision of the radial head, release of the medial collateral ligament (MCL), radial head replacement, and medial collateral ligament reconstruction. METHODS: Twelve fresh human cadaveric elbows were dissected to establish 7 kinds of specimens with elbow joint and ligaments as follow: (1) intact (n=12); (2) release of the medial collateral ligament (n=6); (3) excision of the radial head (n=6); (4) excision of the radial head together with release of the medial collateral ligament (n=12); (5) radial head replacement (n=6); (6) medial collateral ligament reconstruction (n=6); (7) radial head replacement together with medial collateral ligament reconstruction (n=12). Under two-newton-meter valgus torque, and at 0, 30, 60, 90 and 120 degrees of flexion with the forearm in supination, the valgus elbow laxity was quantified: All analysis was performed with SPSS 10.0 software. RESULTS: The least valgus laxity was seen in the intact state and its stability was the best. The laxity increased after resection of the radial head. The laxity was more after release of the medial collateral ligament than after resection of the radial head (P<0.01). The greatest laxity was observed after release of the medial collateral ligament together with resection of the radial head, so its stability was the worst. The laxity of the following implant of the radial head decreased. The laxity of the medial collateral ligament reconstruction was as much as that of the intact ligament (P>0.05). The laxity of the radial head replacement together with medial collateral ligament reconstruction became less. CONCLUSION: The results of this study show that the medial collateral ligament is the primary valgus stabilizer of the elbow and the radial head was a secondary constraint to resist valgus laxity. Both the medial collateral ligament reconstruction and the radial head replacement can restore the stability of elbow. If the radial head replacement can not be carried out, the reconstruction of the medial collateral ligament is acceptable. PMID- 15759934 TI - [Influence of clenbuterol on the sensory neurotrophic activity of denervated red and white muscles]. AB - OBJECTIVE: To investigate the influence of clenbuterol on the expression of nerve growth factor (NGF) in denervated red and white muscles and the neurotrophic activity of the denervated muscles. METHODS: Sixty-four Wistar rats, weighed 200 250 g, were divided into 8 groups (8 rats per group), including 4 experimental groups and 4 control groups. The denervated model was made in rats by dissection of sciatic nerves. Clenbuterol was given at a dose of 200 microg/kg per day in the experimental group, saline in the control group. The expression of NGF was measured with immunohistochemistry after 1, 3, 7 and 14 days of injury. The culture methods of dorsal root ganglions of the chick embryos were used to measure the neurotrophic activity of extracts of the muscles. RESULTS: Compared with the control groups, the NGF content of gastrocnemius (GAS) increased on the 1st day (P<0.05) and the NGF content of soleus (SOL) increased greatly on the 1st, 3rd and 7th day after injury in the experimental groups (P<0.01). In the experimental groups, the NGF amount of GAS reached the highest value on the 1st day after injury (P<0.01) and then decreased gradually. And the NGF amount of SOL had slight difference between different time. The NGF content of the SOL was higher than that of GAS on the 7th day (P<0.05). The sensory neurotrophic activity of the extracts was similar between SOL and GAS. CONCLUSION: Clenbuterol can change the expression of NGF in denervated muscles, but the change was different in SOL and GAS. The sensory neurotrophic activity of the denervated muscles were determined by all of the neurotrophic factors in them. PMID- 15759936 TI - [Advances in gene expression of bone marrow stromal cells]. AB - OBJECTIVE: To review the advances in gene expression of marrow stromal cells (MSCs) in biological characteristic, differentiation, gene therapy, supporting hematopoiesis, inflammation reaction in wound repair. METHODS: The related articles in recent years were extensively reviewed. RESULTS: MSCs can express not only specific mRNA of mesodermal cells but also that of endodermal and ectodermal cell types on various differentiation conditions. When transfected or transduction, MSCs can steadily express many therapeutic transgenes in vitro or in vivo as well. Furthermore, they have the ability to support hematopoietic system and participate in the process of inflammation in wound repair. CONCLUSION: MSCs are stem cells which have the characteristic of self renewing, multi-potency and easy to expand in vitro. MSCs are ideal target cells for cell and gene therapies. PMID- 15759935 TI - [Animal model building of hepaticojejunostomy and hepaticocholedochostomy and comparison of short-term effect]. AB - OBJECTIVE: To discuss the way of animal model building of hepaticocholedochostomy (HC) and hepaticojejunostomy (HJ) and to compare the short-term effect. METHODS: Twenty-nine dogs were divided randomly into control group (n=5) and the experimental group (stenosis of left hepatic duct, n=24). After 7 weeks of stenosis of left hepatic duct, 24 dogs in the experimental group were divided randomly into HC subgroup (n=12) and HJ subgroup (n=12). The operation time and the blood loss during operation were recorded and the hepatic function was detected. RESULTS: The diameter of left hepatic duct was significantly expended after 7 week's stenosis. Hepaticocholedochostomy took shorter time and lost less blood than hepaticojejunostomy. The dogs in HC subgroup lost less weight than those in HJ subgroup. In HC and HJ subgroups, the mortality rates were 1/12 and 3/12; the infectious rates of incision were 3/12 and 5/12 respectively. Serum levels of total bilirubin and transaminase increased significantly in the 7th week after stenosis of left hepatic duct compared with before stenosis of left hepatic duct. However, Serum levels of total bilirubin and transaminase restored to normal levels after 1 month of HC or HJ. CONCLUSION: It is feasible to establish animal model of bile duct reconstruction on the basis of stricture of bile duct. The dogs undergoing hepaticocholedochostomy have less trauma, better results than the dogs undergoing hepaticojejunostomy. Both hepaticocholedochostomy and hepaticojejunostomy are able to relieve the obstruction of bile duct. PMID- 15759937 TI - [Plasticity and application of marrow-derived stem cells]. AB - OBJECTIVE: To investigate plasticity of marrow-derived stem cells, possible mechanisms and application. METHODS: The recent literature concerned plasticity of marrow-derived stem cells was extensively reviewed. RESULTS: Although stem cells plasticity has not been proven, there is more and more evidence to show that the stem cells may have the plasticity and at least four different mechanisms are explained. CONCLUSION: The extensive research will be required. If the plasticity is proven, adult stem cells can be used for more therapeutic purposes. For example, the use of stem cells from bone marrow may correct genetic or degenerative disorders of a second organ from which stem cells are more difficult to isolate. PMID- 15759938 TI - Practice guidelines: friends or foes? PMID- 15759939 TI - Juvenile arthritis: "No child left behind" fits here, too. PMID- 15759940 TI - Clinical diagnostics versus a theoretic algorithm in diagnosing abdominal pain. PMID- 15759941 TI - Impact of national low back pain guidelines on clinical practice. AB - OBJECTIVES: The purpose of this study was to assess the impact of the 1994 Agency for Health Research and Quality (AHRQ) clinical practice guidelines on the management of acute low back pain. METHODS: From the National Ambulatory Medical Care Service database, the authors abstracted data on patients being seen in primary care settings, presenting with low back pain as their primary reason for visit, and aged between 20 and 55 years. Patients with an inflammatory or secondary diagnosis to explain their back pain were excluded. Using the sampling weights assigned by the National Ambulatory Medical Care Service, we assessed the medications prescribed, referrals for physiotherapy, and radiography usage for 3 years before (1991 to 1993) and after (1995 to 1997) release of the back pain guidelines. RESULTS: During these 6 years, more than 10 million ambulatory office visits were available for analysis, 5.2 million visits between 1991 to 1993 and 5.0 million visits between 1995 to 1997. The most common diagnosis was lumbago, present in 21% of these visits. Acetaminophen use increased 20-fold from 0.1 to 2%, nonsteroidal use increased from 40 to 43%, muscle relaxant use decreased from 29 to 20%, radiography ordering increased slightly from 15.4 to 19.4%, and physical therapy referrals declined from 27 to 22%. There was no evidence of a trend toward increased compliance with the AHRQ guidelines over time. CONCLUSIONS: The AHRQ clinical practice guidelines for the management of acute low back pain had a modest impact on physician behavior, increasing the use of acetaminophen and nonsteroidals and decreasing the use of muscle relaxants and physical therapy referrals. PMID- 15759942 TI - Anaerobic spondylodiscitis: case series and systematic review. AB - BACKGROUND: Bacterial spondylodiscitis is rarely caused by anaerobic organisms. We describe two patients with lumbar vertebral osteomyelitis and discitis caused by anaerobic bacteria, including an unusual occurrence after an endodontic procedure, and review the salient clinical features and outcomes of 31 previously reported cases. METHODS: Case reports and review of the literature. RESULTS: Median age at presentation was 65 years, with a male-to-female ratio of 2:1. The most common presenting symptoms were back pain, fever, and neurologic deficits. The lumbar spine was most frequently involved (43%); an equal number of cases involved contiguous extension or hematogenous spread. Causative anaerobes were recovered from disk space or vertebrae (13), blood (4), and/or soft tissue abscess and included Bacteroides species (12), Propionibacterium acnes (7), Peptococcus species (4), Peptostreptococcus species and Clostridium species (3 each), Corynebacterium diphtheroides and Fusobacterium species (2 each), and unspecified anaerobes (3). CONCLUSIONS: Apart from specific antibiotic selection, medical treatment and outcomes for anaerobic spondylodiscitis are similar to those for aerobic vertebral disk infection. PMID- 15759943 TI - Health status in the Mississippi River Delta region. AB - OBJECTIVES: The purpose of the study was to determine if and where there were counties with significantly better or poorer health than others in the region. METHODS: Using Z scores for 6 mortality-related measures, this study identifies counties within the Delta states with the highest and lowest rates. RESULTS: Although Delta counties account for only 33% of the counties in the eight target states, they represent the majority of counties with the highest mortality measures, and represent the minority of counties with positive health outcomes. CONCLUSIONS: These findings confirm the appropriateness of targeting interventions, and help pinpoint places where better health is being achieved despite high poverty and other risk factors, where important lessons can be drawn. PMID- 15759944 TI - Association of clinical and laboratory variables with ultrasound findings in right upper quadrant abdominal pain. AB - BACKGROUND: The purpose of this study was to determine how laboratory values and physical examination findings correlate with ultrasound findings in the setting of right upper quadrant pain. METHODS: Patients undergoing emergent ultrasound for the evaluation of biliary disease between November 1999 and April 2000 were included. Physical examination findings, laboratory data, and ultrasound results were variables. Logistic regression was performed. Ultrasound diagnosis of acute cholecystitis, cholelithiasis, and normal biliary tract were end points. One hundred seventy-seven patients were enrolled. RESULTS: Forty-two percent were diagnosed with acute cholecystitis, 30.5% with cholelithiasis, and 27.1% with normal biliary tract. Alkaline phosphatase, Murphy sign, white blood cell count, and total bilirubin were statistically significant predictors of acute cholecystitis. A Murphy sign was defined as arrest of inspiration with pressure over the right upper quadrant. CONCLUSIONS: The findings from this study allow clinicians to apply objective significance to laboratory data and physical examination findings in patients with suspected gallstone disease. The data can be applied to create a predictive model. PMID- 15759945 TI - Do gastroenterologists notify polyp patients that family members should have screening? AB - OBJECTIVE: The objective of this study was to determine whether patients found to have adenomatous polyps or cancer were notified that their relatives should have screening, due to an increased risk of developing colorectal cancer. METHODS: Consecutive (n = 121) colonoscopy patients from December of 1999 to October of 2001 found to have adenomatous colon polyps or colon cancer formed the study group. Charts were reviewed for documentation of relative notification, and when documentation was not present, study subjects were contacted by telephone. RESULTS: Overall, 71% had data that were able to be evaluated; the remaining 29% were unable to be contacted because of changes of address or phone numbers. Adenomatous polyps were seen in 95%, and cancer seen in 5%. Overall, 30% of the patients were notified: 23 of 82 (28%) in the polyp group and 3 of 4 (75%) in the cancer group. Advanced adenomas or multiple adenomas were noted in 28 of the 82 (34%). Of those, 8 of 28 (29%) were notified. CONCLUSIONS: Gastroenterologists should be aware of the need for increased attention to family notification, especially in those with advanced adenomas or multiple adenomas. Template notification letters may complement the polyp surveillance programs that many colonoscopists use. PMID- 15759946 TI - Daily interferon monotherapy for chronic hepatitis C in patients with normal or near-normal serum alanine transaminase levels: a comparison of two protocols. AB - OBJECTIVES: Up to 30% of the 5 million patients with chronic hepatitis C in the United States have normal serum alanine transaminase (ALT) levels. These individuals have not been treated aggressively, and reported response rates have been low. No study has targeted patients treated in a community practice setting. METHODS: Consecutive patients with chronic hepatitis C with normal or near-normal serum ALT levels seen over a 2-year period in a community gastroenterology hepatology practice were randomly assigned to receive either 3 MU or 6 MU daily interferon (IFN)-alpha-2a monotherapy for 12 months. RESULTS: Sixteen patients (8 in each treatment arm) qualified for study. End-of-treatment response was 87.5% with 6 MU IFN daily and 75% with 3 MU IFN daily, whereas sustained virologic response was 62.5% and 50%, respectively. Genotype 1 patients had an improved outcome with the higher 6 MU dose. CONCLUSIONS: Daily IFN monotherapy achieves high response rates in patients with chronic hepatitis C with normal or near normal ALT. Present-day pegylated interferon regimes can be expected to be as effective. PMID- 15759947 TI - Low utility of endoscopy for suspected upper gastrointestinal bleeding occurring in hospitalized patients. AB - OBJECTIVES: To determine the clinical utility of upper endoscopy in patients who have upper gastrointestinal bleeding after hospitalization. METHODS: Patients were studied who underwent upper endoscopy for an indication of suspected upper gastrointestinal bleeding that developed more than 48 hours after hospitalization. Demographic, clinical, and endoscopic data were extracted by chart review. Bleeding was characterized as clinically important (defined as overt bleeding in association with hemodynamic compromise or the need for blood transfusion) or non-clinically important. RESULTS: Eighty-six patients met inclusion criteria. Clinically important bleeding occurred in 17%. Peptic ulcer disease and gastritis were the most common sources of bleeding in the clinically important and non-clinically important groups, respectively. The bleeding source was not found in 24% of patients. Endoscopic therapy was required in 11% (all of whom had clinically important bleeding). Upper endoscopy prompted no treatment changes in the non-clinically important bleeding group. CONCLUSIONS: Endoscopic therapy was needed only in the few patients with clinically important bleeding. Nonendoscopic treatment can be recommended for upper gastrointestinal bleeding developing in hospitalized patients who do not meet established criteria for a clinically important bleed. PMID- 15759948 TI - Integrative therapy for fibromyalgia: possible strategies for an individualized treatment program. AB - One of the most complex patient treatment situations encountered by the clinician is the patient who presents with the cluster of signs and symptoms that lead to the diagnosis of fibromyalgia syndrome. While physicians focus primarily on pharmacologic treatment, a number of nonpharmacologic modalities have been shown to benefit patients as well. No one therapy is uniformly effective in every patient; treatment programs consisting of a combination of pharmacologic and nonpharmacologic therapies must be individualized to the patient, and the clinician may have to try several different modalities before reaching an optimal improvement in the patient's symptoms. PMID- 15759949 TI - Laboratory testing in the rheumatic diseases: a practical review. AB - Laboratory testing for the rheumatic diseases can allow for rapid diagnosis and appropriate management, while false-positive tests can lead to inappropriate management and unnecessary concern for the patient. An evaluation of laboratory testing for rheumatic illnesses is discussed, including the well-known acute phase proteins, the use of ANA in screening, and the newer antibodies which may potentially allow for an earlier diagnosis. A thorough history and examination are arguably the best screening tests. Clinicians should be judicious in their use of laboratory testing, and should only do so in an attempt to further refine the diagnosis. PMID- 15759950 TI - Role of biological agents in immune-mediated inflammatory diseases. AB - A new era in the treatment of immune-mediated inflammatory disorders has begun with the clinical availability of anticytokine therapy. Biological agents that are currently available include 3 agents that decrease the activity of tumor necrosis factor-alpha (infliximab, adalimumab, etanercept) and an interleukin-1 receptor antagonist (anakinra), with many more in development. Those extraordinarily effective medications are an important addition to our therapeutic armamentarium, and, although originally developed for rheumatoid arthritis and Crohn disease, have been found to be efficacious in the treatment of seronegative spondyloarthropathies (psoriatic arthritis, ankylosing spondylitis) and juvenile rheumatoid arthritis. Their role is currently being defined in other autoimmune disorders such as uveitis, sarcoidosis, interstitial lung disease, vasculitis, inflammatory myopathies, graft-versus-host disease, and Sjogren syndrome. PMID- 15759951 TI - Treatment of osteoarthritis. AB - Osteoarthritis is a most common health problem in population over age 40 and a leading cause of pain and disability in the United States. Treatments of osteoarthritis incorporate combination of nonpharmacologic modalities, pharmacologic agents and surgical procedures. Unfortunately nonpharmacological modalities are underutilized in the management algorithm. In addition to physical and occupational therapy, diet and exercise play an extremely important role, thus patient education in the above areas is of great importance. Pharmacological measures should be examined carefully by physician and the patient weighting its existing risks and benefits. Surgical procedures are generally reserved for patients with severe arthritis who have persistent aim and significantly reduced function. Presently, there are no proven structure/disease-modifying interventions and therefore current therapy is aimed to symptom relief and rehabilitation. PMID- 15759952 TI - Juvenile rheumatoid arthritis: physical therapy and rehabilitation. AB - Juvenile arthritis is one of the most prevalent chronic diseases in the childhood period (ages 0 to 16 years). This disease was first defined in the first half of the 16th century. In the course of time, its differential diagnosis and characteristics have been determined, and it has been classified. Incidence and prevalence values are 10 to 20 in 100,000 and 56 to 113 in 100,000, respectively. Various factors are suggested for its underlying cause. Its denomination is also in dispute. Treatment of juvenile arthritis includes education, medical treatment, physical therapy, and occupational therapy. This article summarizes the objectives and methods of physical therapy and rehabilitation that are important parts of treatment. PMID- 15759953 TI - Ischemic colitis: a clinical review. AB - Ischemic colitis is the most common form of intestinal ischemia. It manifests as a spectrum of injury from transient self-limited ischemia involving the mucosa and submucosa to acute fulminant ischemia with transmural infarction that may progress to necrosis and death. Although there are a variety of causes, the most common mechanism is an acute, self-limited compromise in intestinal blood flow. Patients typically have mild abdominal pain and tenderness over the involved segment of bowel. There is usually passage of blood mixed with stool, but hemodynamically significant bleeding is unusual. Although computed tomography may have suggestive findings, colonoscopy is the procedure of choice for diagnosis. Supportive care with intravenous fluids, optimization of hemodynamic status, avoidance of vasoconstrictive drugs, bowel rest, and empiric antibiotics will produce clinical improvement within 1 to 2 days in most patients. Twenty percent of patients will have development of peritonitis or may deteriorate despite conservative management and will require surgery. PMID- 15759954 TI - Lumbar synovial cyst as a cause of low back pain and acute radiculopathy: a case report. AB - The authors report the case of a 69-year old female with a persistent history of low back pain. Plain lumbar radiographs, performed at symptom initiation, revealed lumbar spondylolisthesis. Low back pain was attributed to this, and the patient received conservative treatment with partial relief of symptoms. Four years later, and as a result of acute onset of radicular symptoms, a lumbar spine magnetic resonance imaging scan was performed, revealing an epidural cystic lesion. The cyst was removed, and histopathology revealed a synovial cyst. PMID- 15759955 TI - Vertebral osteomyelitis caused by Prevotella (Bacteroides) melaninogenicus. AB - A 35-year-old, previously healthy female presented with severe low back pain, fever, and a high erythrocyte sedimentation rate 1 week after a routine dental cleaning. Technetium-labeled leukocyte scanning and magnetic resonance imaging scan of the spine were negative for osteomyelitis. The patient underwent biopsy, cultures from which grew Prevotella (Bacteroides) melaninogenicus. Appropriate antibiotic therapy resulted in resolution of symptoms. P. melaninogenicus is a gram-negative anaerobic bacillus that is part of the indigenous oral flora. It may cause dental, sinus, skin, and soft tissue infections. Infection of bone is rare. Only three cases of vertebral osteomyelitis due to P. melaninogenicus have been reported in the literature. The early diagnosis of vertebral osteomyelitis requires a high index of clinical suspicion and cannot be excluded by negative imaging tests alone. The recovery of this unusual organism highlights the importance of requesting anaerobic cultures of biopsy specimens. PMID- 15759957 TI - Mesenteric venous thrombosis in a patient with pancreatitis and protein C deficiency. AB - Mesenteric vein thrombosis is an uncommon manifestation of hypercoagulable states. A case is reported of superior mesenteric vein (SMV) thrombosis in a patient with pancreatitis and protein C deficiency. A discussion of SMV thrombosis identification, management, and outcomes is included. The patient presented with a significant history of alcohol abuse and constant, midepigastric abdominal pain associated with nausea and vomiting. Amylase and lipase were elevated, and the patient was treated for pancreatitis. His symptoms initially responded to intravenous fluid hydration, but soon his clinical picture worsened, with increased nausea and vomiting, abdominal pain, and distension. Contrasted computed tomography of the abdomen revealed SMV thrombosis. A hypercoagulable workup revealed protein C deficiency. After a 3-month course of oral anticoagulant therapy, the SMV thrombosis resolved. PMID- 15759956 TI - A case of brucella spondylodiscitis with extended, multiple-level involvement. AB - Brucellosis is a zoonosis that affects several organs and has a protean presentation. The authors report the case of a 61-year-old male patient with brucellar spondylodiscitis involving several vertebrae and a paravertebral abscess localized in the erector spinae muscle. Diagnosis was made by positive blood culture and MRI. No relapse was seen with a combined treatment (doxycycline/rifampin) for 3 months, followed by doxycycline alone for 6 months. Almost all radiologic findings disappeared at the end of a 1-year follow-up without any further treatment. PMID- 15759958 TI - Cholesterol emboli-induced renal failure and gastric ulcer after thrombolytic therapy. AB - Cholesterol embolization syndrome is a rare but devastating complication of thrombolysis. Clinical presentations are variable, which has resulted in labeling this syndrome as the great masquerader. Almost every organ in the body may be affected, but the syndrome commonly involves the kidney, skin, central nervous system, and gastrointestinal tract. Treatment is mainly supportive, with an emphasis on reducing the risk of recurrence. The case presented is a unique one of thrombolytic-induced cholesterol embolization syndrome causing renal failure, in which the diagnosis was supported by a biopsy of a gastric ulcer. PMID- 15759959 TI - Hepatic plasmacytosis as a manifestation of relapse in multiple myeloma treated with thalidomide. AB - Thalidomide and its analogs have been extensively studied in patients with multiple myeloma. We present the case of a 58-year-old female patient with immunoglobulin GA-kappa multiple myeloma who was receiving thalidomide after failing an autologous transplant. She presented with profound asthenia and several space-occupying hepatic lesions, one of which was shown by a CT-guided percutaneous biopsy to be plasmacytoma. The patient then received bortezomib and had a transient response. Because thalidomide may also increase the expression of cytoadhesion molecules in myeloma cells and in the bone marrow microenvironment, it is possible that some patients with multiple myeloma who relapse on thalidomide may present with extramedullary plasmacytomas, as seen in this case. Therefore, whenever symptoms arise in patients with multiple myeloma who are receiving thalidomide, extramedullary plasmacytomas should be considered. PMID- 15759961 TI - Celiac sprue presenting as advanced liver disease. AB - This case report describes the complexity of diagnosing celiac sprue in a patient who presented with features predominantly resembling that of advanced liver disease in the absence of diarrhea. PMID- 15759960 TI - Acute liver failure due to iron overdose in an adult. AB - The vast majority of acute iron toxicity cases occur in children less than 5 years of age. Moreover, clinical hepatic injury is uncommon with most symptoms stemming from the intestinal tract (eg, nausea, vomiting, diarrhea). Therefore, physicians, particularly those who do not routinely treat pediatric patients, are often unfamiliar with hepatotoxicity related to iron overdose. Nevertheless, hepatotoxicity caused by acute iron poisoning is associated with a high mortality rate. We report a case of severe hepatic injury in an adult who overdosed on iron tablets with suicidal intent. Tests for other hepatotoxins (eg, acetaminophen), hepatatrophic viruses, and other causes of acute liver injury were negative. Although peak serum iron level (340 microg/dL) was significantly lower than that reported to cause hepatotoxicity (>1,700 microg/dL), rapid and significant elevations in aminotransferases (>4,000 U/L), total bilirubin (5 mg/dL), and prothrombin time (50 seconds) occurred within 48 hours. Treatment with deferoxamine was prompt and followed by empiric N-acetylcysteine once liver injury was apparent. The patient was minimally symptomatic and she eventually had a full recovery. PMID- 15759962 TI - Liver metastasis presenting as pneumoperitoneum. AB - A 63-year-old man presented to the emergency department with sudden-onset abdominal pain. Chest radiography demonstrated pneumoperitoneum. At surgery, the source was found to be a ruptured hepatic abscess. Cultures grew Clostridium perfringens, and biopsies confirmed metastasis of a previously resected pancreatic cancer. We document this rare cause of pneumoperitoneum and briefly review the literature on liver abscess as it relates to metastatic cancer. PMID- 15759963 TI - Cutaneous draining sinus tract of odontogenic origin: unusual presentation of a challenging diagnosis. AB - A 44-year-old woman presented with a chronically draining lesion on her cheek just lateral to the nasofacial sulcus. The lesion was refractory to treatment with oral antibiotics. Physical examination revealed poor dentition, and a panoramic radiograph demonstrated periapical abscesses in the maxillary right lateral incisor and canine. A diagnosis of cutaneous fistula of odontogenic origin was made, and the patient was treated with tooth extraction. The cutaneous fistula subsequently resolved. Intraoral examinations and radiographs are critical for making the diagnosis of cutaneous draining sinus tract of odontogenic origin. Many patients undergo unnecessary surgical therapies before having the correct diagnosis made, but root canal therapy or surgical extraction is the treatment of choice. A dental origin must be considered for any chronically draining sinus of the face or neck. PMID- 15759964 TI - Fatal hypermagnesemia caused by an Epsom salt enema: a case illustration. AB - The authors describe a case of fatal hypermagnesemia caused by an Epsom salt enema. A 7-year-old male presented with cardiac arrest and was found to have a serum magnesium level of 41.2 mg/dL (33.9 mEq/L) after having received an Epsom salt enema earlier that day. The medical history of Epsom salt, the common causes and symptoms of hypermagnesemia, and the treatment of hypermagnesemia are reviewed. The easy availability of magnesium, the subtle initial symptoms of hypermagnesemia, and the need for education about the toxicity of magnesium should be of interest to physicians. PMID- 15759965 TI - Colistin-associated acute renal failure: revisited. PMID- 15759966 TI - Parotid gland involvement in sarcoidosis. PMID- 15759967 TI - Is Japan sufficiently prepared to deal with bioterrorism? PMID- 15759968 TI - The patient's page. Fibromyalgia. PMID- 15759969 TI - [Spectral study of three compounds of tungstosilicic salt with Keggin structure]. AB - Three compounds of tungstosilicic acid and tungstosilicic salt: K3H[SiW(VI)12O40].3H2O(I), (H3O)4[H3SiW(VI)9W(V)3O40].2H2O(II) and [(CH3)4N]4[SiW(VI)12O40].4.5H2O(III), with Keggin structure, have been synthesized. The relationship between their properties and structures was studied by using FTIR, NIR FT-Raman, UV-Vis and flurescence etc. They possess isolated [SiW12O40] cage anion, which combines with cation and water by static electric action and weak hydrogen bond; The study of their FTIR and Raman spectra shows that the vibrational frequencies of the group are related to the structure of the materials. For the compound II, the vibrational frequency of vas(W=Od) decreases as part of W6+ are reduced to W5+, and the charges of anion group affect vas(W-Oc W), vas(Si-Oa) and delta(Oa-Si-Oa) etc. apparently. In UV-Vis spectra of compound I and II, there are two characteristic peaks at 200 and 260 nm, respectively. The fluorescence spectra of compound I were studied, and showed that the emission peak is at 350, 440, 520 and 675 nm excited by 220, 350 and 440 nm. PMID- 15759970 TI - [Spectral studies on nano-sized SO4(2-)/TiO2 photocatalyst prepared by hydrothermal heating the solution of Ti(SO4)2]. AB - In this paper, the nano-sized SO4(2-)/TiO2 solid superacid photocatalysts have been prepared by the method of hydrothermal heating the mixed solution of Ti(S04)2 and urea, heating-decomposing, wet impregnating-calcining. The middle products and the ultimate products have been characterized by XRD, BET, FTIR, DRS and FS. The degradation of rhodamine B by photocatalyst was regarded as a model reaction. The optimum condition of preparing SO4(2-)/TiO2 photocatalysts has been chosen. The results show that hydrothermal treating the solution of Ti(SO4)2 can get the anatase nano-sized TiO2 in the weak alkalinity medium; at 300 degrees C by-products of hydrothermal reaction, such as (NH4)2SO4 and so on, can be decomposed by calcining for 4 hours, avoiding much loss of H2SO4; the most important factors that influence the photocatalystic activities of SO4(2-)/TiO2 are the concentration of SO4(2-) on TiO2 and the calcination time. SO4(2-)/TiO2 photocatalysts have the best photocatalystic activities, up to the level of P-25 photocatalyst, when the concentration of SO4(2-) on TiO2 is 11% and the calcination temperature is 450 degrees C. PMID- 15759971 TI - [Nonlinear optical absorption characteristics of a novel organic material ZnTBP/CA/PhR]. AB - The measurement of optical absorption spectrum in visible light range of a novel organic material Zn-tetrabenzoporphincrotonic acid/phenoxy resin (ZnTBP/CA/PhR) is reported in this paper. The nonlinear absorption property of the sample illuminated by lasers with different wavelengths was studied. And the absorption alteration and kinetic process of the major spectrum bands were observed. The saturation absorption (SA) and reverse saturation absorption (RSA) appeared as the input fluence increased, and re-reverse saturation absorption process of the material was demonstrated in the experiment. These properties occurred in a broad range of visible spectrum. The paper qualitatively analyses the physical mechanism during the whole course. Based on the nonlinear absorption characteristics, we showed some applications in photonics field of ZnTBP/CA/PhR. PMID- 15759972 TI - [Solid phase extraction and room temperature phosphorimetry coupling technique and its application]. AB - The principle and application in the environmental analysis of Solid Phase Extraction (SPE) combined with Room Temperature Phosphorescence (RTP) analysis were reviewed. The principle, device, separation modes, operation procedures and application of SPE were commented. The merits and shortages in the operation procedure and quantitative analysis of classic RTP were also summarized. The potential application in the trace sample analysis and the advantage of coupling technique of SPE and RTP were indicated. The sensitivity and selectivity of the coupling technique could be greatly increased because of the strong enrichment ability of the SPE materials and the high selectivity of RTP. So far the technique has been successfully used in the analysis of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PClDBFs) in the environmental samples according to their phosphorescence emission after preconcentration on the Whatman 1PS filter paper and C18 or C8 SPE membranes. PMID- 15759973 TI - [Studies on photoluminescence characteristics of curcumin in acrylamide polymer]. AB - A photoluminescence polymer was prepared using acrylamide as a monomer, a large excess of ethyleneglycol dimethacrylate as the cross-linking agent, and curcumin as a fluorescence additive. The polymer was ground into powder and screened in order to get even powder, whose diameter was from 40 to 75 microm. The emission spectra of curcumin in solid powder polymer show a strong blue shift compared with that of curcumin in C2H5OH solution. Meanwhile, rhodamine 6G, tetraiodofluorescein and phloxin have the same phenomenon of blue shift. Based on these facts, we speculate that the mechanism is probably singlemolecular luminescence because of the limit effect in three dimension space of the polymer. The molecule fluorescence peak was also depending on its environment because the surrounding electron field affected the molecule luminescence in polymer. PMID- 15759974 TI - [Chemiluminescence method for determination of dipyridamole]. AB - A new system for the determination of dipyridamole with flow injection chemiluminescence was described. It is based on the chemiluminescence reaction of dipyridamole-potassium permanganate with rhodamine B. Tween-80 was found to be an enhancer of the chemiluminescence reaction. A method based on the enhanced chemiluminescence for dipyridamole determination has been developed. The method has high sensitivity, selectivity and good repeatability with a linear concentration range of 5.0 x 10(-8) - 5.0 x 10(-5) g x mL(-1), a detection limit of 1.7 x 10(-8) g x mL(-1) and a RSD of 1.1% (n = 11, cs = 1.0 x 10(-6) g x mL( 1)). PMID- 15759975 TI - [Electron spectra of chemical assembly mesoporous MCM-41 with transition metal complexes]. AB - In the paper the complexes of metal cobalt(III), manganese(III), iron(III) and copper(II) with Schiff-base N,N-ethylenebis(salicylideneaminnato) have been synthesized and characterized. The nanosized porous material MCM-41 has been functionalized by the modification of the internal pore surface with gamma aminopropyl and was assembled by the modification groups with metal complex of Schiff-base. These metal complexes and functionalized nanosized porous materials were charcterized by XRD, IR and UV-Vis. It is indicated by spectral analysis that the synthesized nanosized porous materials have been confirmed to be MCM-41 with hexagon bores, and the gamma-aminopropyls have been bonded on their internal pore surface. And the complexes of metal cobalt(III), manganese(III), iron(III) and copper(II) with Schiff-base N,N-ethylenebis(salicylideneaminnato) were assembled into MCM-41. The IR spectra of these samples show that there were characteristic absorptions of the amino and the Schiff base groups, and the absorption of amino shifted to longer wavelength. PMID- 15759976 TI - [Approximate calculation of L-S coupling of aluminum 3p electronic angular momentums]. AB - Al plasma was obtained by a pulsed laser beam, which was produced from a Nd:YAG laser set with Q-switch, ablating metal target Al in Ar. Based on the plasma, radiation of the plasma was recorded with time- and space-resolved technology, so Al characteristic radiation spectra were obtained. According to the spectra, the coupling of aluminum 3p electronic angular momentums was simplified as quasi hydrogen particle. At the same time, the main quantum and orbit angular quantum shielding coefficients of aluminum nucleus, signed as alpha and s respectively, were carefully figured out with related theories of quantum mechanics. With our experiment, the resonance double lines of aluminum were scaled up at 396.15 and 394.40 nm respectively. As a result of our calculation, the main quantum shielding coefficient of aluminum nucleus was sigma = 10.850359, and the orbit angular quantum one was s = 7.332875. Their values were reasonable in terms of the order of magnitude. PMID- 15759977 TI - [N II ion spectra related to lightning discharges]. AB - Appling the atomic structure theory to the research field of physical processes of lightning discharges, parameters such as wavelengths, oscillator strengths, transition probabilities and excitation energy of upper levels have been calculated for the transitions of N II ions related to lightning discharges. Large-scale multi-configuration Dirac-Fock wavefunctions were applied to include the most important effects of relativity, correlation, and rearrangement of the electron density within the same (computational) model. More detailed identification than ever for lightning spectra has been done, and reference data have been provided for further theoretic and experimental works on the physical mechanism of lightning discharges. PMID- 15759978 TI - [Study on the spectroscopy of paramagnetic compounds [{Cu(bipy) [C6H4(COO)2]}2.H2O]n and [Cu(phen) [C6H4(COO)2]]n]. AB - The cluster compounds with d9-d9 magnetic system were investigated because of their magnetic character. Some of the copper(II) compounds with terephthalato as bridged unit, [{Cu(bipy) [C6H4(COO)2]}2.H2O]n and [Cu(phen) [C6H4(COO)2]]n, have been synthesized through a solvothermal reaction, and their crystal structures have been determined by X-ray diffraction. Their shape looks like zigzag-chain. In this paper, the molecular vibration spectra of the title complexes have been studied in detail. The middle and far-infrared spectra, and Raman spectra of the compounds have been reported and their assignments have been made experientially. The characteristic carbonyl absorption of terephthalato group at 1688 cm(-1) disappeared while the complexes formed. The characteristic peaks of COO- group were splited into two peaks respectively that appeared at 1603, 1574 cm(-1) (vasCOO) and 1369, 1356 cm(-1) (vsCOO) in [{Cu(bipy)[C6H4(COO)2]}2.H2O]n, and at 1614, 1605 cm(-1) (vasCOO) and 1373, 1344 cm(-1) (vsCOO) in [Cu(phen) [C6H4(COO)2]]n. The relationship between the structures of the compounds and the spectra has been discussed as well. The coincidence between experiential assignments and the theoretic calculation will be confirmed. PMID- 15759979 TI - [Time-resolved FTIR study of amorphous process of nylon 6 with calcium chloride]. AB - The influence of calcium chloride on the crystallinity of nylon 6 was studied by means of time-resolved FTIR, WADX and DSC. The results show that the crystallinity of nylon 6 decreased with the increase of CaCl2, and finally nylon 6 changed from crystal phase to amorphous phase. Because hydrogen bond is the main reason that nylon has three-dimensional ordered structure and forms crystalline phase, we further studied the changes of wave number of NH, amide I and amide II related to hydrogen bond in CaCl2/PA6 complexes by time-resolved FTIR. These studies indicate that in the chloroethanol solution of nylon 6 and calcium chloride, complex bonds between calcium cation and oxygen atom of carbonyl group gradually form, and calcium cations are inserted into the molecular chains of nylon 6, which weaken the hydrogen bonds between the molecular chains of nylon 6 and damage the three-dimensional ordered structure of nylon 6, so the crystallinity of nylon 6 decreases and nylon 6 changes from crystalline to amorphous phase. PMID- 15759980 TI - [In situ raman spectroscopic study on the oxidation of HCHO on a roughened platinum electrode]. AB - The electrooxidation behavior of HCHO on a roughened platinum electrode was studied by cyclic voltammetry. Two factors that influence the electrooxidation behavior of HCHO, i.e. the concentration of the supporting electrolyte and the structure of the electrode surface were taken into account. The dissociative adsorption behavior of HCHO on a roughened platinum electrode was investigated by confocal microprobe Raman spectroscopy in-situ, and the spontaneous dissociative adsorption behavior of HCHO on a roughened platinum electrode was found by CV and was confirmed at the molecule level by in-situ Raman spectra. PMID- 15759981 TI - [Infrared absorption spectroscopy of optical waveguide in proton-exchanged LiNbO3]. AB - Z-cut proton-exchanged LiNbO3 optical waveguides exchanged with different proton sources mixed with adipic acid and benzoic acid in different mol ratios were analyzed by infrared absorption spectroscopy. The result shows that there is an absorption peak of OH- free group at 3500 cm(-1) with the characteristic of substitute protons, and the absorption peak of H-bond is nearby 700 cm(-1) with the characteristic of packing protons. The different mixing ratios and different concentrations of H+ in the proton source, result in different proton-exchanged velocities and bring on the change in these absorption peaks. It is very important to study the change in these absorption peaks for making high quality LiNbO3 optical waveguide. PMID- 15759982 TI - [Preparation and properties of complex of europium lanthanum with crown ether and p-phthalic acid]. AB - Two complexes of crown ether europium (EuL) and crown ether lanthanum (LaL) were synthesized by europium perchlorate and lanthanum perchlorate with 18-crown-6(18 C-6) respectively. The heternuclear complexes of europium-lanthanum p-phthalic acid crown ether were prepared by the two complexes (EuL and LaL) with p-phthalic acid. The elemental analysis shows that its composition is EuLaL2L'(ClO4)4.6H2O(L = 18-C-6 and L' = p-C6H4 (COO)2). The IR spectra, UV-spectra and molar conductor indicate that both ligands (L and L') coordinate rare earth ions (Eu3+ and La3+), but perchlorate group is free. The fluorescence spectra show that the fluorescence intensities of the complex of heternuclear (EuLaL2L'(ClO4)4.6H2O) are more than eight times that of the crown ether europium complex. It was shown that the linked ligand (p-phthalic acid) can transfer the absorbed energy of Lanthanum-crown ether to europium-crown ether with high efficiency. PMID- 15759983 TI - [Identification of gastrodia elata blume by Fourier transform infrared spectroscopy]. AB - In this paper, a method of rapid and undamaged identification of wild and cultivated Gastrodia elata Blume, and one of its fakes by Fourier transform infrared spectroscopy (FTIR) is reported. The results show that Gastrodia elata Blume and its fake have different characteristic infrared spectra, by which Gastrodia elata Blume can be identified from its fake. Wild winter, wild spring, and cultivated Gastrodia elata Blume can be discriminated by FTIR, according to the differences of their spectral peaks and absorbance ratios. By the differences of absorbance ratios of several peaks, different grade of Gastrodia elata Blume may be classified. FTIR has proved to be a rapid, simple and nondestructive method for the identification of Gastrodia elata Blume. PMID- 15759984 TI - [Study on the identification of dangguitou and dangguiwei by two-dimensional infrared correlation spectroscopy]. AB - Fourier transform infrared spectroscopy (FTIR) combined with two-dimensional correlation spectroscopy was used to identify the different parts of Danggui in this paper. Guitou and Guiwei are two parts of the same officinal plant, and similar to each other in normal. FTIR spectra revealed the comparabilities of these two parts in herbal characters, organizing structure and alkaloid components. However, variance in amino acids and naphtha contents, which is aroused by the different parts of the angelica, could be identified easily and clearly in 2D FTIR spectra. Thus the fast and accurate property of 2D FTIR spectroscopy makes it a powerful and new approach to evaluating the medicinal herbs impersonally. PMID- 15759985 TI - [FTIR of adipose tissue of Ming dynasty mummy]. AB - The Fourier transform infrared spectra (FTIR) of adipose tissue of Ming dynasty GUCONGLI mummy were investigated by chloroform-methanol extraction and two ways of infrared sample preparation. The results show that the triglycide in the adipose tissue was hydrolyzed into free fatty acids, and both the octdecenic acid and saturated fatty acids existed. PMID- 15759986 TI - [Application and progress of Fourier transform attenuated total reflection infrared spectroscopy]. AB - With the application of Fourier transform infrared spectrograph and the development of chemical metrology, Fourier Transform Attenuated Total Reflection Infrared spectroscopy (ATR-FTIR) has become a kind of beneficial tool and means for analyzing samples, for which traditional transmission way does not work quite effectively, and for analyzing surface layer structure. ATR-FTIR has been applied to every realm such as spinning and weaving, quality testing, public security and so on. At present, people are launching applied study based on specific property of ATR-FTIR. Therefore, this paper mainly introduced the present study situation and development situation of ATR-FTIR, including theoretical development for depth profiling, qualitative analysis of material surface, quantitative analysis of mixture and solution component, and the application of fiber as ATR appurtenance. In addition, some special fields were also involved, such as inspecting hollow cored fiber's structure and change of orientation, and studying effective mechanism about skin promoter. PMID- 15759987 TI - [Detection of organic autotrophic particles by FCM (flow cytometry) in an oligotrophic lake]. AB - Organic autotrophic particles were detected by flow cytometry (FCM) in an oligotrophic lake. Utilizing scattering signals and fluorescence of their natural pigments, different populations could be rapidly discriminated and enumerated. We found that they belonged to three populations. R1 were the smallest population, while R3 were the largest one. R1 contained phycoerythrin but lacked chlorophyll. On the contrary, R3 contained a large amount of chlorophyll but lacked phycoerythrin. R2 contained not only chlorophyll but also a great deal of phycoerythrin. Organic autotrophic particles account for the most proportion of the total particles that were smaller than 50 microm. They represented 70.24% of the total particles. In this study, we found that flow cytometry is particularly suited to the analysis of organic autotrophic particles that are difficult to study with traditional methods. PMID- 15759988 TI - [Study of the effect of aliphatic alcohol on the inclusion interaction of qucertin-cyclodextrin complexes]. AB - The inclusion interaction of qucertin with beta-cyclodextrin and its derivatives was studied by fluorimetry. The formation constants of their inclusion complexes were determined, and the stoichiometry ratio on the formation of the inclusion complexes was determined by equimolar continueous variation method. The effect of aliphatic alcohols with short chains on inclusion process was discussed. The results showed that the inclusion ability of beta-CD and its derivatives was in the order: HP-beta-CD>M-beta-CD>beta-CD. The effect of aliphatic alcohol on complexes was related to the carbon number and volume of alcohol molecules. PMID- 15759989 TI - [Synthesis of mixed complexes of lanthanum with 2,2-bipyridyl and 1,10 phenanthroline and study of the interaction of the complexes with DNA]. AB - A mixed complex of La with 2,2-bipyridyl (dpy) and 1,10-phenanthroline(phen) was synthesized. Elemental analysis, IR and TGA of the complex were investigated, thus it was proved that the conformation of it was La(dpy)(phen)Cl3. The interaction of the complex with DNA has been studied also. The spectra showed that the interaction between La(dpy)(phen)Cl3 and DNA experienced intercalation of long-range self-assembly. The study made it clear that the synthesized mixed complex La(dpy)(phen)Cl3 can interact with DNA, and La(dpy)(phen)Cl3 can be used for probing a nucleic acid. PMID- 15759990 TI - [Color reaction of new tailed porphyrin-pyridine quaternary ammonium salts with copper (II)]. AB - Color reactions of Cu2+ with three new different-chain-length tailed porphyrin pyridine quaternary ammonium salts were studied. The Soret bands of complexes lie at (I), (II) 413 nm and (III) 414 nm, which are separated from those of the diacid forms (I) 443.5 nm, (II) 444 nm and (III) 446 nm. The contrast is about 30 nm. The ratio of Cu2+: reagent in the complexes is 1:1. The apparent molar absorptivities of complexes are (I) 3.4 x 10(5), (II) 2.9 x 10(5) and (III) 2.5 x 10(5) L x mol(-1) x cm(-1), and Beer's law is obeyed in the ranges of (I)0-0.5 microg x (10 mL)(-1), (II) 0-0.6 microg x (10 mL)(-1) and (III) 0-1 .0 microg x (10 mL)(-1) for Cu2+. This method can be used for the determination of trace Cu2+. PMID- 15759992 TI - [Synthesis and spectral characterization of boron complexes of bis-schiff base from acylpyrazolone]. AB - Three novel bis-schiff base complexes [B(PMaFP)2en]Ac, [B(PMaFP)2pen]Ac and [B(PMTHP)2en]Ac have been synthesized, where (HPMalphaFP)2 en = N,N'-bis [(1 phenyl-3-methyl-5-oxo-4-pyrazolinyl) alpha-furylmethylidyne] ethylenediimine, (HPMalphaFP)2pen = N,N'-bis[(1-phenyl-3-methyl-5-oxo-4-pyrazolinyl) alpha furylmethylidyne]-o-phenylenediimine and (HPMTHP)2en = N,N'-bis[(1-phenyl-3 methyl-5-oxo-4-pyrazolinyl)-2-thenoylmethylidyne] ethylenediimine. They were characterized by elemental analysis, IR, UV-Vis, 1H NMR, 13C NMR, and molar conductance measurements. The results show that the bis-schiff base is a quadridentate ligand and boron is four-coordinated in the complexes. PMID- 15759991 TI - [The reformation of tomato ecology using photo-ecology film]. AB - The action spectrum of photosynthesis of tomato was studied with excitation and fluorescence spectra of its leaf. To simulate action spectrum of tomato the fluorescent auxiliary was specially prepared and used to make the photo-ecology film. The film was used to cover the roof of greenhouses for planting tomatoes in Jianguo village of Xinfa town, Daoli district, Harbin. The survey of ecology conditions, tomato growth and yield has been made in comparison with those in the greenhouse using normal film. It is shown that the yield of tomato in the photo ecology film greenhouse has increased by 53.7%, which leads to a production value rise of 61.4%. PMID- 15759993 TI - [Study on the fluorescence spectra of LaL3 with DNA]. AB - A fluorescence method is presented for the determination of DNA. The method is based on the interaction of LaL3 [L = morin (2', 3, 4', 5, 7 Pentahydroxyflavone), 2'-OH group deprotonated] with DNA in NH3 x H2O-NH4Cl buffer of pH = 8.0, with lambdaex at 387 nm, and lambdaem at 535 nm. Enhanced fluorescence was observed for LaL3 in the presence of DNA and in the presence of buffer solution of 10%(psi) and ethanol of 10%(psi). The linear range of determination was between 0 and 15 microg x mL(-1) 1 for DNA. The method proved to be simple, easy and sensitive for the determination of DNA. The mechanism of enhancing and the reasons for the effects of acid were also discussed. PMID- 15759994 TI - [Catalysis-fluorescence determination of trace nitrite based on the oxidation of acridine orange by potassium bromate]. AB - Acridine orange can be oxidized with discoloring and fluorescence quenching by potassium bromate in phosphoric acid medium, but the reaction rate is very low. The presence of trace nitrite can catalyze this reaction to improve the sensitivity of it apparently and to make the net fluorescence value of the system decrease greatly. Based on the above investigation, a new catalysis-fluorescence determination method of trace nitrite has been proposed. The method described is very sensitive and selective. The catalysis reaction is under the conditions of lambdaex/lambdaem = 446/505 nm, 55 degrees C and 10 min for the reaction time. The method permits the existence of more than 20 common foreign ions. There isn't interference by the same quantity of Fe3+, Br- and I-. The detection limit is 0.012 ng x mL(-1). The linear range of determination is 0.05-5 ng x mL(-1). The method has been used for the determination of trace nitrite in water and soil samples with satisfactory results. PMID- 15759995 TI - [Ion floatation-spectrophotometric determination of palladium(II) content in acetaldehyde catalyst]. AB - The nature of palladium (I) and copper (II) is very similar, so it is very difficult to determine trace palladium (II) in the coexistence of a great quantity of copper (II). This paper applied ion floatation-spectrophotometric principle, using 1-(2-pyridylazo)-2-naphthol (PAN) as collector, and through controlling pH of the solution and determination wavelength, without separation to determine palladium (II) content in acetaldehyde catalyst with a great quantity of copper (II). The relative error was 0.61%, and RSD was 2.33%. The results were satisfactory. PMID- 15759996 TI - [Study on the method for determination of sulfate in Fe-catalyst by inductively coupled plasma atomic emission spectrometry]. AB - A direct method was reported for the determination of sulfate in iron-catalyst by inductively coupled plasma atomic emission spectrometry (ICP-AES). The samples were dissolved with hydrochloric acid. The interference factors of hydrochloric acid and iron were studied. The results showed that the effect of iron on the test of sulfate was not only to cause background interference but also to depress the intensity of sulfur seriously as well, and the effect increased with the increment of iron amount. Due to the physical in terference, a high concentration of HCl also depresesed the intensity of sulfur. Matrix effect from iron and HCl were corrected by matrix matching method. The recommended method is simple and rapid, and show satisfying precision and accuracy. The recoveries were more than 97%, and the relative standard deviations of six samples were less than 3%. PMID- 15759997 TI - [Determination of trace selenium in coal by microwave digestion and hydride generation ICP-AES]. AB - A method for determining trace Se in coal by microwave digestion and hydride generation ICP-AES is reported. The digestion system of HNO3-HCl-HF was evaluated. The hydride generation occurred in 3 mol x L(-1) hydrochloric acid media with 1% sodium tetrahydroborate as a reduction reagent. The experimental conditions of microwave digestion, prereduction, and interferences were investigated. This method is simple and rapid. The treatment method was applicable to Se and gave results that were in good agreement with the certified values of the standard reference material. Testing results showed that the detection limit was 0.3 microg x L(-1) and RSD was below 5%. PMID- 15759998 TI - [Determination of micro-elements in couplers by inductively coupled plasma atomic emission spectrometry]. AB - By ICP-AES, we established a method to quantitatively determine the micro elements in couplers. We processed the unknown sample with wet digesting method. Nine micro-elements in the couplers can be quantitatively determined at the same time. Using the powerful chem.-station, we have discussed the method to eliminate physical interference and background correction. The results of the experiment showed that the method is accurate, quick, of high performance, and with all RSDs < or = 1.00%. The linear correlation coefficient of the tested elements is over 0.9999. PMID- 15759999 TI - [Determination of trace elements in flowers and tests of conditions]. AB - The optimum conditions, such as the ratio of fuel gas to combustion-supporting gas, burnerheight, lamp current and band pass, for the determination of trace elements such as calcium, magnesium, cobalt and potassium in flowers were studied by flame atomic absorption spectrophotometry (AAS). The experimental results showed that Beer's law was obeyed in the range of 0-50 microg x mL(-1) for Ca, 0 80 microg x mL(-1) for Mg, 0-4 microg x mL(-1) for Co, and 0-120 microg x mL(-1) for K. The correlation coefficients were in the range of 0.9975-0.9995 under the seleced conditions. Applied to the determination in rose & Chinese rose the method proved to be simple, rapid and selective with satisfactory recovery of 97% 104% and relative standard devition of 1.2%-2.3%. PMID- 15760000 TI - [Analysis of the contents of K, Na, Ca, Mg, Zn, Cu, Fe and Mn in serum of middle and old-aged hypertension patients]. AB - Atomic absorption method was used to detect the contents of K, Na, Ca, Mg, Zn, Cu, Fe and Mn in serum of 48 hypertension patients and 52 healthy people. The results show that the contents of Na, Mg, Zn, Fe and Mn in hypertension were higher than healthy control, while K, Ca and Cu were lower. Na/K were higher, but Ca/Mg and Cu/Zn were lower, than healthy control. These suggested that we should add K, Ca, Mg and Cu properly, reduce Na, scientifically control the increase of Zn, Fe and Mn, pay attention to the balance relation of Na/K, Ca/Mg and Cu/Zn, reduce Na/K, raise Cu/Zn, and add Ca with Mg properly. This can be useful for preventing and treating hypertension. PMID- 15760001 TI - [Determination of zinc in mineral by FAAS after phase separation and extraction]. AB - Non-ion surface active agent can be separated from its solution of water by heating to its cloud point. A new method for the determination of zinc in mineral samples by flame atomic absorption spectrophotometer after phase separation and extraction making use of its cloud point was studied in this paper. The conditions of phase separation and extraction such as using Triton X-100 as phase separation agent, and octanol to drop the cloud point, and the influence of acidity and the number of times for oscillating were discussed. The detection limit was 0.005 microg x mL(-1). This method has been applied to the determination of zinc in ores with satisfactory results. PMID- 15760002 TI - [Determination of trace elements in shenrong heixi wan]. AB - Shenrong Heixi Wan is a pure traditional Jiangxi medicine which has been mainly used to cure coma in clinic as its main function is resuscitation. In this paper, four trace elements in Shenrong Heixi Wan and other five traditional Chinese medicines have been determined and contrasted by AAS. The results show that there are abundant benificial trace elements in Shenrong Heixi Wan while pernicious trace elements such as Hg, Cd etc. are low. So the medicine is safe to take. PMID- 15760003 TI - [Studies on sequential injection spectrophotometric system using a reflective flow cell for process analysis]. AB - A reflective flow cell was developed and coupled to a sequential injection system and optical fiber photometric detection system based on emitting diode light source. A multistrand bifurcated optical fiber was coupled with incident light, detector and flow cell. Optical fibers were used to carry light from the electronics unit to a reflective flow-through cell and back. The liquid flow path through the cell is linear with a large exit aperture such that bubbles are not trapped in the optical path. The optical arrangement is such that the incident light crosses the liquid flow orthogonally and is reflected back to the receiver fiber. This arrangement reduces the reflective index sensitivity by an order of magnitude relative to a conventional flow cell. The cell showed good immunity to refractive index and air bubble effects. The chromogenic reaction of chloride ion with mercury thiocyanate-iron(III) was used as a model reaction to optimize the experiment system and check the optical system. The reflectance of the reaction was monitored with blue emitting diode. The linear range was 0-100 mg x L(-1) Cl . A detection limit (3sigma) of 1.2 mg x L(-1), precision of 1.5% (n = 11), and a throughput of 30 samples per hour were achieved. PMID- 15760004 TI - [The depolarization characteristic of quartz depolarizer in UV-VUV spectral region]. AB - The quartz depolarizer consisting of two wedge-shaped quartz crystals was designed based on the Lyot depolarizer. The quartz depolarizer is used to reduce the polarization response of spectral instruments. The residual degree of polarization of the quartz depolarizer was derived by the method of matrix optics, so the variation regularity of its depolarization characteristic was analysed concerning its center thickness and the status of the incident radiation in the UV-VUV spectral range. Its depolarization effect was obvious for the radiation with some spectral bandwidth. Also, the depolarization characteristic of the quartz depolarizer was studied experimentally with 1-meter Seya-Namioka UV VUV spectrometer, using LiF crystals as the polarizer and analyzer. The residual degree of polarization of the quartz depolarizer was less than 1%. Both the theoretical analysis and experimental results show that the quartz depolarizer we designed is appropriate to the spectral measurement with high accuracy. PMID- 15760005 TI - [Mathematical model of dispersive infrared gas analyzer based on pyroelectric detector]. AB - This paper analyzes the characteristics of the pyroelectric detector based on its working principle. The input andoutput mathematical model of DIGA (Dispersive Infrared Gas Analyzer) system with pyroelectric detector was established according to the design principle of DIGA. We have manufactured a novel multi-gas DIGA on the basis of this model, then pointed out several problems that should be taken into account in the design. Application indicates that this model is of considerable practical value for the design, study, performance analysis and further improvement of DIGA. PMID- 15760006 TI - [Study on the phosphorylaions reaction between ethyl salicylate and DEPH]. AB - Atherton-Todd reaction has been extensively applied to the synthesis of phosphates and phosphoroamidates. Under Atherton-Todd reaction conditions, dialkyl phosphites were probably transformed into diakyl phosphorochloridates. We have tried to use Atherton-Todd reaction to synthesize phosphate of salicylic acid. But when the -COOH of salicylic acid was under unprotected, the yield was quite low. It was found that the -COOH possibly participated in the reaction. The competitive reaction between the -COOH and -OH of salicylic acid probably existed. In order to understand the detail mechanism, ethyl-salicylate was selected to be phosphorylated In the end, ethyl-salicylate successfully phosphorylated by modification of the classical Atherton-Todd procedure through dropping the DEPH and tetrachloromethane into the mixed solution of ethyl salicylate, triethylamine and dioxane with good yields. The structures were elucidated by NMR and ESI/MS/MS data. PMID- 15760007 TI - [Coordination bonding driving supramolecular self-assembly of tetraphenylporphyrinatozinc (II) with imidazolyl tailed porphyrinatoiron (III) studied by fluorescence spectra and 1H NMR]. AB - Coordination bonding driving supramolecular self-assembly of Zn(TPP) with Fe(p ImEPTPP)Cl was investigated by fluorescence spectra and 1H NMR. The results indicate that an obvious fluorescence quenching observed from Zn(TPP) by addition of Fe (p-ImEPTPP)Cl is due to the coordination of the terminal imidazolyl group of Fe(p-ImEQTPP)Cl to Zn2+ of Zn(TPP). The change of fluorescence spectra of Zn(TPP)/Fe(p-ImEPTPP)Cl system by adding some Lewis bases, such as piperidine, imidazole and pyridine, as well as 1H NMR further supported this coordination. PMID- 15760008 TI - [Measurement and analysis of optical constants of bulk ZnSe in THz region]. AB - In the present paper the authors measured and analyzed the dispersion and absorption characteristics of the bulk ZnSe(110) in THz region using transmission THz time domain spectroscopy. After fitting the experiment data according to the Drude theory, the authors obtained the low-frequency dielectric constant and the high-frequency dielectric constant. The authors also observed the absorption by multi-phonon processes. PMID- 15760009 TI - [Crystal growth and spectral parameter computation of Yb:YAG]. AB - In this article, a good-quality crystal Yb:YAG was grown by pull method, and its absorption spectrum of 200-3000 nm was measured at room temperature. Its absorption of 200-300 nm is from the host YAG, and there exists only the characteristic absorption of Yb3+ in the range of 390-3000 nm. Judd-Ofelt theory computation indicates that the absorption and emission oscillator strengths of electric dipole of (24 at%) Yb:YAG are 3.58 x 10(-6) and 4.77 x 10(-6), respectively, and the absorption and emission transition probabilities are 879 and 1171/s respectively. As for magnetic dipole, the absorption and emission oscillator strengths are 3.32 x 10(-7) and 4.43 x 10(-7) respectively, and the absorption and emission transition probabilities are 82 and 109/s respectively. The lifetime of the energy level2 F(5/2) is 781 micros. PMID- 15760010 TI - [Preparation of Ge/SiO2 nanometer-crystal doped glass and study of its photoluminescence property]. AB - Ge/SiO2 nanometer-crystal doped glass was prepared through sol-gel method. The gel was synthesized through the hydrolysis of a complex solution of Si (OC2H5)4 and Ge(OC2H5)4. The product gel was then heated at 700 degrees C in H2. Ge crystals appeared after the heating-reducing process. X ray diffraction analysis showed that there were Ge cubic crystals formed in the gel glass. The photoluminescence spectroscopic analysis showed that the Ge/SiO2 fluorescence peaked at 576 nm with excitation wavelength of 494 nm. By using the Brus quantum confinement model, the fluorescence peak energy suggested that the mean size of Ge crystal was about 3 nm. PMID- 15760011 TI - [Spectral studies of nanosized LnVO4:Eu (Ln = La, Gd, Y)]. AB - Nanosized LnVO4:Eu (Ln = La, Gd, Y) with diameters ranging from 20 to 50 nm were synthesized by complex sol-gel technique. The structures and spectral properties of LnVO4:Eu were studied by X-ray powder diffraction (XRD), Fourier transform (FT IR) spectroscopy, luminescence spectra and ultraviolet spectroscopy (UV). The experimental results show that YVO4:Eu and GdVO4:Eu have similar spectral properties because they both have the structure of xenotime, while the structure of LaVO4:Eu belongs to monazite, whose symmetry is inferior to the former vanadates. Consequently the spectral properties of LaVO4:Eu are obviously different from the other two luminescence materials, as the peaks of infrared spectrum and emission spectrum become wider and the number of peaks in UV-Vis spectrum increases. PMID- 15760012 TI - [The luminescence characteristic of a new novel coprecipitate rare earths complexes]. AB - A new novel coprecipitate rare earths complexes La0.6Eu0.4(BSA)3phen has been synthesized and was chosen as the emitter material in the organic electroluminescent devices: ITO/PVK: La0.6Eu0.4(BSA)3phen/Alq/Al where PVK was used to improve the film-forming and hole-transporting property of the La0.6Eu0.4(BSA)3phen. It has been proved that it exists Forster energy transfer process from La3+ to Eu3+. This device has also been compared with the devices: ITO/PVK: Eu(BSA)3phen/Alq/Al and ITO/PVK: Tb0.6Eu0.4(BSA)3phen/Alq/Al. From the data, it indicated the device has the good purity of red color and good commutation property. The maximum EL brightness of 102 cd x m(-2) had been got. PMID- 15760013 TI - [Electroluminescence property of MEH-PPV/ZnO nano-crystal structure]. AB - A new structure with II-VI nano-crystal ZnO film as the electron transport layer and MEH-PPV as the hole transport layer and emitting layer was fabricated. The new structure's brightness and efficiency were improved markedly compared with that of the single MEH-PPV layer structure. And a new peak at 620 nm was found in the ITO/MEH-PPV/ZnO/Al structure's electroluminescence spectrum, which should resulted from ZnO layer. In addition, the turning-on voltage of the two-layer structure decreased from 9 V of the single layer structure to 4 V. From the I-V curve, the authors concluded that the emitting area was around the interface of MEH-PPV/ZnO and the combination area could shift with changing the voltage. PMID- 15760016 TI - [Resonance-enhanced multiphoton ionization spectrum of NO by D2 sigma state]. AB - NO is an important pollutant molecule, and plays a key role in generating photochemical smog and destroying O3 in the atmosphere. So researchers have always been interested in it. In our experiment, the optical parameter generator and amplifier (EKSPLA) pumped by Nd:YAG (Quantel) laser were used as the radiation source. The REMPI spectrum of NO in the region of 460-570 nm was obtained. Based on theoretical calculation, most of the lines were attributed to the X2 pi --> D2 sigma transition. The ionization pathway was discussed in detail, which can be shown as NO(X2 pi) (3h nu)--> NO(D2 sigma) (2h nu)--> NO+ + e. The vibration fundamental and force constant of NO (D2 sigma) state were calculated. The results are omega(e) = 2339.2 cm(-1) and k = 2.41 x 10(5) dyn x cm(-1). For future work, the resolution of the radiation source will be improved in order to obtain more information of the D2 sigma state. PMID- 15760015 TI - [Research of spectrum characteristics for light conversion agricultural films]. AB - The solar spectrum and the function spectrum in chrysanthemum and tomato were determined in this paper. The research for a relation plant growth to solar spectrum showed that the efficiency of plant making use of ultraviolet light of 280-380 nm and yellow-green light of 500-600 nm and near IR spectra over 720 nm are lower, that the blue-purple light of 430-480 nm and red light of 630-690 nm are beneficial to enhancing photosynthesis and promoting plant growth. According to plant photosynthesis and solar spectrum characteristic, the author developed CaS:Cu+, Cl- blue light film, and red light film added with CaS:Eu2+, Mn2+, Cl- to convert green light into red light, and discussed the spectrum characteristic of red-blue double peak in agricultural film and rare earth organic complex which could convert ultraviolet light into red light. Just now, the study on light conversion regents in farm films is going to face new breakthrough and the technology of anti-stocks displacement to study red film which can convert near infrared light are worth to attention. PMID- 15760017 TI - [Phase separation and transport behaviors in La(0.7 - x)Dy(x)Sr0.3MnO3 system]. AB - The influence of Dy doping (0.00 < or = x < or = 0.30) on magnetic and electric properties of La0.7Sr0.3MnO3 was studied by measuring M-T curves, M-H curves, ESR curves, rho-T curves and MR-T curves of the samples. The experimental results indicate that all samples undergo the transition from paramagnetism to ferromagnetism at T(C). When T < T(C), the doped samples enter the cluster-spin glass state, and the antiferromagnetism is exhibited at lower temperature. For the samples x = 0.2, the phase separation occurs at the temperature far above T(C), the variation in magnetic structure caused by Dy doping leads to colossal magnetoresistance. PMID- 15760018 TI - [Temperature distribution measurement of high energetic monopropellant by spectroscopic diagnostic technology]. AB - The combustion flame temperature distribution in the axes are measured by relative intensity method of the spectroscopic diagnostic technology for monopropellant hexanitrohexaazaisowurtzitane (HNIW) at 3 MPa and 5 MPa pressure, respectively. The investigation results show that the curves of combustion flame temperature distribution in all combustion course are accurately measured by relative intensity method for monopropellant HNIW. The measured highest combustion flame temperature in the axes are lower than theoretical combustion temperature at the same pressure, and are more close to theoretical combustion temperature along with the rising of the pressure. The experimental results indicate that the combustion flame temperature distribution can be measured by the relative intensity method for high energetic and high burning rate propellant at higher pressure. PMID- 15760019 TI - [Application of molecular absorption spectrophotometric method to the determination of biologic macromolecular structures]. AB - A review on recent applications of molecular absorption spectrophotometric method to the identification of the structures of biologic macromolecules, such as protein and nucleic acid, is presented. Molecular absorption spectrophotometric method is widely used in the structure determination of biologic macromolecules for its convenience and speed. Ultraviolet absorption spectrum (UV) can be used in the research on the mechanism of the interaction of anticancer drugs and DNA. UV can also be used to study the interaction of spectroscopic probe with biologic molecule and their binding mechanism. Protein secondary structure and conformation can be investigated by Fourier infrared spectroscopy (FTIR) deconvolution analysis. Molecular absorption spectrophotometric method is an important tool for structure study of biologic macromolecules. PMID- 15760020 TI - [Investigation on the detection of pesticide residue in vegetable based on infrared spectroscopy]. AB - In this paper, the mid-infrared Attenuated Total Reflection (ATR) spectra of two slathered pesticides dichlorvos and trichlorfon in vegetable juice solution have been investigated. It can be concluded that within characteristic absorption region of the two pesticides in the mid-infrared range, the pigments inside the vegetable have no effect on the pesticide' s absorbance; pesticides in standard solution and vegetable juice solution share almost the same absorbance characteristics. These results indicate that: the authors can use the model built by the absorbance data of pesticides in water solution to simulate their absorbance in vegetable solution, then based upon infrared spectroscopy, the direct detection of pesticide residue on the vegetable can be achieved; it also provides a possible way of rapid detection on vegetable in the future. PMID- 15760021 TI - [Outlier diagnosis and calibration model optimization for near infrared spectroscopy analysis]. AB - Outlier diagnosis is a very important step in building near infrared calibration model. Data outlier includes spectral outlier and chemical value outlier. Mahalanobis' distance, ratio of spectral residual and spectral variable leverage test were used to evaluate sample spectral outlier. Cook's distance and the ratio of sample square error of chemical value and predict value to the mean square error of calibration set were used to test chemical value outlier. Three calibration models of protein content of 50 wheat samples, protein content of 90 corn samples and cyclohexane content of four compounds mixture were investigated. It is demonstrated that outlier test is very helpful for optimizing near infrared calibration model. PMID- 15760022 TI - [The analysis of Gynostemma pentaphyllum (Thunb) Makino for Chinese crude drug by FTIR and FT-Raman]. AB - The Gynostemma pentaphyllum is an important plant for Chinese crude drug. To qualitatively analyse the Gynostemma pentaphyllum (Thunb) Makino, FTIR and FT Raman were used to identify the contents of it. The results show, that the trough of each absorption in the picture of FTIR spectrum and the characteristic wave crest of each dispersion in the picture of FT-Raman spectrum had parallelism at their position, their relation pairs with each other. This is useful for the further on-line quantitative analysis and quality control of the Gynostemma pentaphyllum (Thunb) Makino. This approach provide a quickly and accurately determination method of crude drug without extraction and separation. PMID- 15760023 TI - [Study on the resonance scattering light spectra of atrazine-RNA and its analytical application]. AB - The Resonance Light Scattering (RLS) spectra of atrazine with yeast ribonucleic acid (yRNA) had been studied. At pH 1.50, the enhanced RLS of atrazine-yRNA system is characterized by the maximum scattering peaks at 320 nm and the RLS intensity is proportional to the concentration of yRNA. Under optimal conditions, the linear ranges of the calibration were 0.6-5.0 microg x mL(-1), the linear regression equation was I = 25.88 + 140.0 c(yRNA, microg x mL(-1)), r = 0.9975, with the limit of detection 20.7 ng x mL(-1) (3delta). The method was applied to the determination of yRNA in synthetic samples and RNA in salt cress (Thellungiella halophila) sample satisfactorily. Mechanism studies show that there are two interaction modes between atrazine and RNA: electrostatic mode and intercalative mode. PMID- 15760025 TI - [Study of photocatalytic activity, characterization and preparation of supersuspended nano-TiO2. II. Study of photocatalytic activity]. AB - From the change in the absorption of methyl-orange solution at 470 nm, the photocatalytic activity of nano-TiO2 prepared by pyrolysis method, and then by cooling treatment in silicon oil and in air respectively, was studied with methyl orange solution as model waste water under irradiation by UV-lamp. Their photocatalytic mechanisms, the percentage of degradation in 35 min and the kinetic reactive process were also analyzed. The results show that the photocatalytic activity of both products is obvious with little difference. However, the nano-TiO2 powder modified with silicone oil can degrade harmful components more effectively on the surface of waste water with sunlight and other extra light. PMID- 15760024 TI - [Catalytic application of synthesizing n-butyl acrylate by a new type nanometer complex heteropoly acid catalyst H3PW12O40/SiO2]. AB - A new nanometer complex heteropoly acid with Keggin structure, H3PW12O40/SiO2, were prepared by sol-gel method, and were characterized with IR, UV, XRD and TEM techniques. By means of this nanometer catalytic materials, the optimum conditions of the n-butyl acrylate synthesis have been studied. The results show that the complex heteropoly acid H3PW12O40/SiO2 nanoparticles have the mean grain size of 40 nm and they are typical amorphous. A strong chemical interaction exists between H3PW12O40 and silica surface. The nanoparticles have high catalytic activity for synthesizing n-butyl acrylate. The optimum catalytic conditions are as follows: the mole ratio of acrylic acid and n-butyl alcohol is 1:1.2, the reaction temperature is approximately 90-96 degrees C, and the catalyst quantity in the reaction is 10% of the acid mass. The conversion proportion is 94.37% and product yield 91.2% in 5 h. Apparently, the unique structure of the Keggin anions and surface acid center and the high specific surface area and the pseudoliquid phase of H3PW12O40/SiO2 play an important role in the esterification reactions with the acid catalyst. PMID- 15760027 TI - [Research on discrimination of 3D fluorescence spectra of phytoplanktons]. AB - Six phytoplankton species belonging to four divisions, which may cause red tide, were selected and cultivated under different temperature (20 degrees C and 15 degrees C) and illumination (7000, 4100 and 1100 Lux) in lab. 3D fluorescence spectra were measured in each growth period. Discrimination of these phytoplanktons' 3D fluorescence spectra was explored using discriminant analyses. The result showed that discriminant analyses based on the principal component analysis of spectra data was an efficient method. Isochrysis galbana, Platymonas helgolanidica and Skeletonema costatuma can be discriminated effectively. Skeletonema costatuma, which is an diatom, can be distinguished from Alexandrium tamarense and Prorocentrum micans which belong to Dinophyta. PMID- 15760026 TI - [Effect of CHAPS on the stability of Spirulina platensis phycobilisomes]. AB - The Spirulina platensis phycobilisomes were isolated by sucrose density gradients ultracentrifugation, and the fluorescence emission maximum of the phycobilisomes at room temperature was at 671 nm. The effects of ionic strength and the zwitterionic detergent CHAPS on the stability of the Spirulina platensis phycobilisomes were studied by room temperature fluorescence spectrum. The phycobilisomes were stable in 1.0 mol x L(-1) phosphate buffer solution, and their fluorescence emission maximum could remain unchanged for 7 days. The fluorescence emission maximum of phycobilisomes was blue-shifted to 648 nm when the concentration of the phosphate buffer solution was diluted to 0.1 mol x L(-1) with deionized water, which suggested that the phycobilisomes had been dissociated. The phycobilisomes were readily dissociated in phosphate buffer solutions of low concentrations (< 0.6 mol x L(-1)) and the speed of the dissociation increased with decreasing the concentration of the phosphate buffer solution. The fluorescence emission maximum of the phycobilisomes in 1.0 mol x L( 1) phosphate buffer solution was blue-shifted to 648 nm when 10 mmol x L(-1) CHAPS was added into the phycobilisomes solution, suggesting that CHAPS could dissociate phycobilisomes under high ionic strength conditions. The results might be useful for isolating intact substructures of phycobilisomes. PMID- 15760028 TI - [Studies on a new fluorescence-enhanced system of nucleic acids-morin-Al(III)]. AB - A fluorescence-enhanced system was developed for the determination of nucleic acids by using morin-aluminum(III) complex as a new fluorescent probe. In aqueous solution, morin-aluminum(III) complex showed maximum excitation and emission wavelengths at 420.0 nm and 532.8 nm, respectively, and its fluorescence could be greatly enhanced in the presence of nucleic acids. Under optimal conditions, the calibration graph was linear over the range 0.25-1.50 microg x mL(-1) for fish sperm DNA, 0.10-1.60 microg x mL(-1) for salmon sperm DNA, 0.25-2.00 microg x mL( 1) for calf thymus DNA and 0.25-2.00 microg x mL(-1) for yeast RNA. The corresponding detection limits are 3 ng x mL(-1), 2 ng x mL(-1), 2 ng x mL(-1) and 3 ng x mL(-1), respectively. Applied for the determination of nucleic acids in synthetic samples, the relative standard deviation for five replicates is less than 3.6%, and the recovery ranges from 93.3% to 107.9%. Additionally, the interaction mechanism of morin-aluminum(III) with nucleic acids is also discussed. PMID- 15760029 TI - [Determination of free chlorine with spectrophotometric method using N,N-diethyl 1,4-phenylenediamine]. AB - In this paper, the absorption spectra of free chlorine in the visible region has been measured after free chlorine in the solution had reacted with N,N-diethyl 1,4-phenylenediamine (DPD). The relation between the absorption spectra and peak wavelength has been discussed. From the experiment of liquor with different concentration, the error of spectral measurement is less than 0.04%. By nonlinear data fitting, the relation between the concentration of free chlorine and the transmittivity has been given. PMID- 15760030 TI - [Study on the simultaneous spectrophotometric determination of three components including deoxyschizandrin by ridge regression method]. AB - Ridge Regression (RR) spectrophotometry was used in the present paper to analyse three components: deoxyschizandin, schisandrin, and gamma-schisandrin. The basic principle and the analytical steps of the approach are described in detail. The computer program of RR is based on VB language. The experimental results show that the RR method has no systematical error as compared to classical method, and the average recovery of each component is all in the range of 92.35% to 108.69%. High accuracy of RR method was obtained. Each component was determined with satisfactory results without any pre-separation. As compared with conventional methods, this method is simple, rapid and suitable for the computer-aided analysis. It was developed in statistical literature to treat these ill conditioned systems and is an useful analytical method. PMID- 15760031 TI - [Molecular recognitions of purines by hematoporphyrin and metalloporphyrin receptors]. AB - The present paper studies the interactions of purines and porphyrin by fluorescence, UV, and NMR spectra. The optimal conditions (acidity and ion intensity) of the interaction of purines and hematoporphyrin and metalloporphyrin were investigated in detail. The result shows that hematoporphyrin and metalloporphyrin exhibit significant recognition of purines. The binding constants of metalloporphyrin: K(Ni(II)-HP) > K(Co(II)-HP) > K(Zn(II)-HP) > K(Cu(II)-HP) in pH 11.2 Kolthoff buffer solution. The mechanism of recognition was further discussed. Recognition is achieved by cooperative functions of three recognition groups (metalloporphyrin) and two recognition groups (hematoporphyrin). PMID- 15760032 TI - [Measurement of uranium isotope ratio in solid sample by laser ablation and double-beam diode laser atomic absorption]. AB - The 235U/238U isotope ratio was determined by diode laser atomic absorption spectrometry in a plasma induced by a Nd:YAG laser. Two diode lasers, which probed almost the same plasma volume, were used as light sources. 235U and 238U absorption signals were simultaneously measured by tuning the diode lasers to the absorption lines 682.6736 nm and 682.0768 nm, respectively. The optimal focus position for the Nd:YAG laser was found to be 0.4 cm below the sample surface. The precision and accuracy of 235U/238U isotope ratio determination were obtained to be 5% and 2%, respectively. The detection limit of the 235U isotope, evaluated on the basis of 3sigma criteria, was estimated to be 18 microg x g(-1). All these results show an improvement compared with the method in which only one diode laser was used as a light source. PMID- 15760033 TI - [Uncertainty in-statistical-monitor for ICP-AES measurement system by using moving range pooled]. AB - This paper describes how to estimate the uncertainty when Fe in metallic silicon is determined by ICP-AES (Inductively Coupled Plasma Atomic Emission Spectrometry). The technique of the averaged moving range (MR) pooled in statistical-dynamic-monitor is proposed. The study suggests that the uncertainty contribution of ICP-AES measurement system is, by using this technique, a combination of quality control (QC) and linear fit. This conclusion maximized the combination of the effects on various variances, refrained from the complicated relativity, and favored the uncertainty evaluation for ICP-AES measurement system. PMID- 15760034 TI - [Application of orthogonal test in determination of trace elements in wild vegetables by ICP-AES]. AB - In this paper, orthogonal test was applied to determination of trace elements in wild vegetables just as Mn, Se, Cu, Fe, Mg and Zn by ICP-AES. It could provide many informations for us by less experimental times, and could choice the best experimental conditions by square-error analysis. It would be helpful for improving experimental accuracy. The method was simple, rapid, and possessed of highly sensitivity, good accuracy and determination of many elements at the same time, in addition little environment pollution. The recoveries of the method were in the range of 97.2%-103.3%, and relative standard deviations were less than 2.15%. The experimental results proved wild vegetables were valuable and nutritious food, they had rich trace elements. PMID- 15760035 TI - [Determination of main components in Nd-Fe-B magnetic materials by ICP-AES]. AB - The method for direct determination of neodymium, iron and boron in Nd-Fe-B permanent magnetic materials by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) was studied and suitable analytical lines were selected. The interference from matrix elements in samples was diminished by matrix matching method. The memory effect of the determination of boron by ICP-AES was discussed and was eliminated by way of detecting and subtracting blank value from testing result of samples frequently. The method was applied to the determination of Nd, Fe and B in Nd-Fe-B permanent magnetic materials samples with a precision of RSD 0.62%, 0.56% and 0.90%, respectively. PMID- 15760036 TI - [Study on the determination of calcium in Chinese medicine by ICP-AES]. AB - Calcium is the necessity element in human body and its content has effect on exciting muscle or nerve. Calcium is important in the physiologic process of muscle or nervous reaction, nervous impulse transmission, heartbeat rhythm maintenance, blood solidification, cell coherence and so on. Determination of calcium element in Chinese medicine wild jujube seed decoction and radix puerariae was studied by ICP-AES after they were digested with HNO3-HClO4. The precision (RSD) is 1.7%. The average recovery of calcium in wild jujube seed decoction is 97.7% and in radix puerariae is 104.4%. Calcium was abundant in Chinese medicine and could play certain pharmacologic role in Chinese medicine. It is of important significance to assess the value of Chinese medicine by determination of calcium element in Chinese medicine with ICP-AES. PMID- 15760037 TI - [Study on the determination Cu, Cr, Al, Ni and Ti in corrosion solution of oxidation equipment in supercritical water by ICP]. AB - The Cu, Cr, Al, Ni and Ti in corrosion solution of oxidation equipment in supercritical water by inductively coupled plasma-atomic emission spectrometry has been investigated, the authors used catalysts (CuO/Al2O3) to oxidize unsymmetrical dimetylhydrazine in supercritical water in a stainless steel (1Cr18Ni9Ti) reactor. The test was operated under condition of 400-500 degrees C and 24-26 MPa. The results indicate that the dissolution of Cr in stainless steel were occurred in supercritical water and dissolution became severe with higher temperature and pressure. PMID- 15760038 TI - [Determination of mercury and selenium in bone by atomic absorption spectrometry after microwave digestion]. AB - Mercury and selenium in the rats' thighbone were determinated by cold atom absorbance and flow injection hydride atom absorbance after digesting by microwave. The method of sample's making and digesting was discussed. The factors of determination of selenium were studied. The detection limits of mercury and selenium are 1.65 and 1.78 ng x mL(-1) respectively. The RSD% of mercury and selenium are 4.1% and 3.6% respectively. The linearity of calibration curve of mercury and selenium are in the concentrations of 0-120 ng x mL(-1) and 0-50 ng x mL(-1) respectively. The recovery of mercury and selenium are 93.72%-101.8% and 95.53%-102.2% respectively. PMID- 15760039 TI - [Direct determination of trace arsenic in coal by slurry-sampling graphite furnace atomic absorption spectrometry]. AB - A method for direct determination of trace arsenic in coal by slurry-sampling Graphite-Furnace Atomic Absorption Spectrometry (GFAAS) was developed, using Pd as a matrix modifier. The effects of slurry stability, particle size of sample, matrix modifiers, ashing temperature, atomization temperature, and common coexisting components on the signal intensity of arsenic were investigated. Under optimized operating conditions, the detection limits and relative standard deviation of this method were 0.054 microg x g(-1) and 8.9%, respectively. PMID- 15760041 TI - [Determination of LF-VD refining furnace slag by X ray fluorescence spectrometry]. AB - Eight components, i.e. TFe, CaO, MgO, Al2O3, SiO2, TiO2, MnO and P2O5 in refining furnace slag were determined by X ray fluorescence spectrometer. Because the content of CaO was high, the authors selected 12 national and departmental grade slag standard samples and prepared a series of synthetic standard samples by adding spectrally pure reagents to them. The calibration curve is suitable to the sample analysis of CaO, MgO and SiO2 with widely varying range. Meanwhile, the points on the curve are even. The samples were prepared at high temperature by adding Li2B4O7 as flux. The experiments for the selection of the sample preparation conditions about strip reagents, melting temperature and dulition ratio were carried out. The matrix effects on absorption and enhancement were corrected by means of PH model and theoretical alpha coefficient. Moreover, the precision and accuracy experiments were performed. In comparison with chemical analysis method, the quantitative analytical results for each component are satisfactory. The method has proven rapid, precise and simple. PMID- 15760040 TI - [Determination of trace gold by flow injection on-line preconcentration flame atomic absorption spectrometry with trioctylamine levextrel resin]. AB - A new method for the determination of trace amount of gold by flow injection on line preconcentration and flame atomic absorption spectrometry with trioctylamine levextrel resin was developed. The experimental condition of Au(III) on the resin, such as the time of preconcentration, the flow rate, the elutropic time, the elutropic rate, the acidity of medium, the concentration and acidity of eluting agent was studied in this paper. The detection limit is 1.3 microg x L( 1). The enhancement factor of 24 was achieved for a loading period of 90 seconds. The recoveries of Au were 98.0%-102.0%. The developed method has been applied to the determination of trace amount of gold in water samples with satisfactory results. PMID- 15760042 TI - [Development of portable LED-based NIR integrity wheat component measuring apparatus]. AB - A integrity wheat component quickly measuring apparatus which based on NIR LED was developed, by using new chip and designing software. While providing a lower cost apparatus, it also provides quick answer speed and smaller size for fieldwork of wheat's components measuring. The apparatus is made up of NIR LED, interference filter, lens, Si photronic detector, microprocessor system and calibration model. Compared with other instrument, the apparatus has many advantages, such as compact space, simple structure, low power waste, and anti shake. This paper introduces the design of the apparatus, and predicts the concentration of wheat protein. The apparatus can realize non-damage measurement of wheat components' concentration on fieldwork. PMID- 15760043 TI - Permeability classification of representative fluoroquinolones by a cell culture method. AB - This study was undertaken to categorize representative fluoroquinolone drug substance permeability based on the methods outlined in the Food and Drug Administration's biopharmaceutic classification system (BCS) Guidance for Industry. The permeability of ciprofloxacin, levofloxacin, lomefloxacin, and ofloxacin was measured in an in vitro Caco-2 assay with previously demonstrated method suitability. The permeability class and efflux potential were ascertained by comparing test drug results with standard compounds (metoprolol, atenolol, labetalol, and rhodamine-123). All 4 quinolones drugs demonstrated concentration dependent permeability, indicating active drug transport. In comparing absorptive versus secretive in vitro transport, the tested fluoroquinolones were found to be subject to efflux in varying degrees (ciprofloxacin > lomefloxacin > rhodamine 123 > levofloxacin > ofloxacin). Based on comparison to labetalol, the high permeability internal standard, ciprofloxacin was classified as a low permeability drug, whereas lomefloxacin, levofloxacin, and ofloxacin were classified as high permeability drugs. The in vitro permeability results matched human in vivo data based on absolute bioavailabilities. This laboratory exercise demonstrated the applicability of an in vitro permeability method for classifying drugs as outlined in the BCS Guidance. PMID- 15760044 TI - Acute and subchronic (28-day) oral toxicity study in rats fed with novel surfactants. AB - The toxicity of 2 new synthetic lipids, 1,2-dioleoyl-rac-glycerol-3 dodecaethylene glycol, GDO-12 (lipid 1) and 1,2-distearoyl-rac-glycerol-3 dodecaethylene glycol, GDS-12 (lipid 2) has been evaluated in acute and subchronic toxicity studies. Acute oral toxicity studies in male and female rats documented no deaths or treatment-related signs at high doses. The lipids were individually administered (by gavage) to male and female Sprague-Dawley rats at concentrations of 250, 500, and 1000 mg/Kg bodyweight for 28 days. All animals survived the duration of the study, with no significant changes in clinical signs, hematological parameters, organ weights, ophthalmology evaluations, or histopathological findings. These studies establish that both GDO-12 (lipid 1) and GDS-12 (lipid 2) are nontoxic in rats following oral administration. The no observed-adverse-effect level ranged between 250 mg/Kg and 1000 mg/Kg following oral administration. PMID- 15760045 TI - Evaluation of the potential use of poly(ethylene oxide) as tablet- and extrudate forming material. AB - The purpose of this study was to assess the potential use of poly(ethylene oxide) (PEO) as matrix-forming material for tablets and extrudates. Raw materials were characterized for size, size distribution, and shape. Tablets with 2- and 10-mm diameter were prepared by direct compression at both 13 and 38 MPa from mixtures with poly(ethylene oxide)s, a model drug (propranolol hydrochloride), and lactose. To these mixtures water was added (16%-43%) prior to extrusion in a ram extruder fit with different dies (1-, 3-, 6-, and 9-mm diameter and 4-mm length). Tablets and extrudates were characterized for work of compression or extrusion, respectively, relaxation, tensile strength, friability, and drug release. Both PEOs produced tablets easily and with different properties. Some relaxation was observed, particularly for tablets with higher amounts of PEOs. Release of the drug occurred after swelling of the matrix, and between 10% and 70% drug released, a quasi zero-order release was observed for large tablets. Extrusion was possible for formulations with PEO only with amounts of water between 16% and 50%. Both radial and axial relaxation of both plugs and extrudates were observed. Moreover, different extrusion profiles reflected the different behaviors of the different formulations. The model drug was released in the same fashion as observed for the tablets. It was possible to produce tablets by direct compression and extrudates or pellets from those extrudates from different formulations with PEO. Tablets and pellets have shown distinct properties depending upon the PEO considered. Extrusion was particularly complex with different formulations with PEO. PMID- 15760046 TI - Evaluation of a new coprocessed compound based on lactose and maize starch for tablet formulation. AB - The development of new direct compression excipients should include a comprehensive and rapid determination of deformation properties. The aim of this study was to characterize StarLac, a new coprocessed compound for direct compression based on lactose and maize starch. For this purpose, the effects of the base materials (maize starch and spray-dried lactose) were considered and the influence of the spray-drying process was investigated. This was performed by comparing the physical mixture of starch and spray-dried lactose at the same ratio as for StarLac. For analysis of the deformation behavior, the 3-D model and the Walker equation were applied; for verification, the Heckel equation and the pressure time function (a modified Weibull equation) were used. The advantages of StarLac are its good flowability depending on the spray-drying process, an acceptable crushing force due to its lactose content, its rapid disintegration depending on starch, and a brilliant fast release of an active ingredient, such as theophylline monohydrate. The volume-pressure deformation properties of StarLac were dependent on the lactose properties. Only at high maximum relative density (rho(rel, max)) did the influence of starch cause a change in these properties. A network-like structure can be observed using scanning electron microscopy pictures. Overall, StarLac deformed plastically with a low portion of elasticity. The physical mixture exhibited a more elastic behavior than StarLac. However, the part of the powder that was irreversibly compressed was much lower than was observed for the single substances. This behavior is caused by an interaction between the components, which in StarLac is prevented by spray drying. PMID- 15760048 TI - Biodegradable microspheres as carriers for native superoxide dismutase and catalase delivery. AB - The purpose of this research was to encapsulate superoxide dismutase (SOD) and catalase (CAT) in biodegradable microspheres (MS) to obtain suitable sustained protein delivery. A modified water/oil/water double emulsion method was used for poly(D,L-lactide-co-glycolide) (PLGA) and poly(D,L-lactide) PLA MS preparation co encapsulating mannitol, trehalose, and PEG400 for protein stabilization. Size, morphology, porosity, mass loss, mass balance, in vitro release and in vitro activity were assessed by using BCA protein assay, scanning electron microscopy, BET surface area, and particle-sizing techniques. In vitro activity retention within MS was evaluated by nicotinammide adenine dinucleotide oxidation and H2O2 consumption assays. SOD encapsulation efficiency resulted in 30% to 34% for PLA MS and up to 51% for PLGA MS, whereas CAT encapsulation was 34% and 45% for PLGA and PLA MS, respectively. All MS were spherical with a smooth surface and low porosity. Particle mean diameters ranged from 10 to 17 mum. CAT release was prolonged, but the results were incomplete for both PLA and PLGA MS, whereas SOD was completely released from PLGA MS in a sustained manner after 2 months. CAT results were less stable and showed a stronger interaction than SOD with the polymers. Mass loss and mass balance correlated well with the release profiles. SOD and CAT in vitro activity was preserved in all the preparations, and SOD was better stabilized in PLGA MS. PLGA MS can be useful for SOD delivery in its native form and is promising as a new depot system. PMID- 15760049 TI - Characterization of reservoir-type microcapsules made by the solvent exchange method. AB - The purpose of this research was to characterize and optimize the properties of microcapsules produced by the solvent exchange method, a new microencapsulation technique. Reservoir-type microcapsules containing lysozyme as a model protein were produced using a coaxial ultrasonic atomizer under various formulation and instrument settings, and characterized with respect to in vitro release kinetics and stability of the encapsulated protein. The solvent exchange method could encapsulate protein drugs with high efficiency under an optimized condition and was mild enough to preserve the integrity of the encapsulated lysozyme during the process. In vitro release studies showed that the microcapsules could release proteins in a controllable manner. The solvent exchange method is a mild and simple microencapsulation method that could encapsulate lysozyme, maintaining its functional integrity. PMID- 15760050 TI - Particle size and temperature effect on the physical stability of PLGA nanospheres and microspheres containing Bodipy. AB - The purpose of this study was to investigate the effect of particle size, storage temperature, and duration of storage on the physical stability and morphology of polylactic-co-glycolic acid (PLGA) nanospheres and microspheres. PLGA nanospheres and microspheres containing the fluorescent dye, Bodipy, were prepared in varying sizes by controlling the method and degree of agitation during the emulsification phase of preparation. Mean diameters of the particles were measured by dynamic light scattering. To evaluate the effect of storage temperature and duration of storage on the extent of aggregation, nanospheres and microspheres were stored at 4 degrees C, 25 degrees C, 37 degrees C, and 50 degrees C for 6 days and then monitored using both confocal and scanning electron microscopy. The mean +/- SD diameters of PLGA particles containing Bodipy were: 266.9 +/- 2.8, 351.6 +/- 1.8, 988.8 +/- 14.1, and 1865.9 +/- 67.0 nm. The extent of aggregation of the particulate delivery system decreased as the mean diameter increased, and increased as the storage temperature increased. The maximum extent of aggregation was observed with the smallest (266 nm) nanospheres. Microspheres did not aggregate. The aggregation of nanospheres was significantly reduced by introducing an additional evaporation step during preparation, suggesting that migration of residual dichloromethane from within the nanospheres may have dissolved the PLGA on the surface. The extent of aggregation of nanospheres increased as the temperature was increased from 4 degrees C to 50 degrees C, and decreased as particle size increased. To avoid aggregation, PLGA nanospheres should be stored at 4 degrees C. PMID- 15760051 TI - Hydrophilic excipients modulate the time lag of time-controlled disintegrating press-coated tablets. AB - An oral press-coated tablet was developed by means of direct compression to achieve the time-controlled disintegrating or rupturing function with a distinct predetermined lag time. This press-coated tablet containing sodium diclofenac in the inner core was formulated with an outer shell by different weight ratios of hydrophobic polymer of micronized ethylcellulose (EC) powder and hydrophilic excipients such as spray-dried lactose (SDL) or hydroxypropyl methylcellulose (HPMC). The effect of the formulation of an outer shell comprising both hydrophobic polymer and hydrophilic excipients on the time lag of drug release was investigated. The release profile of the press-coated tablet exhibited a time period without drug release (time lag) followed by a rapid and complete release phase, in which the outer shell ruptured or broke into 2 halves. The lag phase was markedly dependent on the weight ratios of EC/SDL or EC/HPMC in the outer shell. Different time lags of the press-coated tablets from 1.0 to 16.3 hours could be modulated by changing the type and amount of the excipients. A semilogarithmic plot of the time lag of the tablet against the weight ratios of EC/SDL or EC/HPMC in the outer shell demonstrated a good linear relationship, with r = 0.976 and r = 0.982, respectively. The predetermined time lag prior to the drug release from a press-coated tablet prepared by using a micronized EC as a retarding coating shell can be adequately scheduled with the addition of hydrophilic excipients according to the time or site requirements. PMID- 15760052 TI - Agglomeration of Ibuprofen with talc by novel crystallo-co-agglomeration technique. AB - The purpose of this research work was to obtain directly compressible agglomerates of ibuprofen with talc by a novel crystallo-co-agglomeration (CCA) technique, which is an extension of spherical crystallization. Ibuprofen-talc agglomerates were prepared using dichloromethane (DCM)-water as the crystallization system. DCM acted as a good solvent for ibuprofen as well as a bridging liquid for agglomeration of crystallized drug with talc. The agglomerates were characterized by differential scanning calorimetry, powder X ray diffraction, and scanning electron microscopy and were evaluated for tableting properties and for drug release. The process yielded spherical agglomerates containing ~95% to 96% wt/wt of ibuprofen. Agglomerates containing talc showed uniform distribution of hydroxypropylmethylcellulose and decreased crystallinity, and deformed under pressure. The miniscular form of ibuprofen and the hydrophobicity of talc governed the drug release rate. The batch containing a higher proportion of talc showed zero-order kinetics and drug release was extended up to 13 hours. The CCA technique developed in this study is suitable for obtaining agglomerates of drug with talc as an excipient. PMID- 15760053 TI - Stabilized nanoparticles of phytosterol by rapid expansion from supercritical solution into aqueous solution. AB - The basic objective of this work was to form stable suspensions of submicron particles of phytosterol, a water-insoluble drug, by rapid expansion of supercritical solution into aqueous solution (RESSAS). A supercritical phytosterol/CO2 mixture was expanded into an aqueous surfactant solution. In these experiments 4 different surfactants were used to impede growth and agglomeration of the submicron particles resulting from collisions in the free jet. The concentration of the drug in the aqueous surfactant solution was determined by high-performance liquid chromatography, while the size of the stabilized particles was measured by dynamic light scattering. Submicron phytosterol particles (<500 nm) were stabilized and in most cases a bimodal particle size distribution was obtained. Depending on surfactant and concentration of the surfactant solution, suspensions with drug concentrations up to 17 g/dm3 could be achieved, which is 2 orders of magnitude higher than the equilibrium solubility of phytosterol. Long-term stability studies indicate modest particle growth over 12 months. Thus, the results demonstrate that RESSAS can be a promising process for stabilizing submicron particles in aqueous solutions. PMID- 15760054 TI - Investigation of various factors affecting encapsulation on the in-cap automatic capsule-filling machine. AB - The purpose of this study was to determine the factors that influence fill weight and weight variability of capsules produced on the In-Cap and to assess any differences in terms of capsule defects between gelatin and HPMC (Quali-V) shells. The In-Cap is an automatic tamping type capsule-filling machine and the low output of approximately 3000 capsules/hour makes it ideal for early formulation development and phase I/IIa clinical supplies manufacture. Four commonly used excipients (Avicel PH101, Avicel PH302, A-Tab, and Prosolv HD90) and a poorly flowing drug blend were encapsulated at various pin settings and powder bed heights. The average fill weight and coefficient of weight variation were determined. The percentage of defective capsules formed during encapsulation was calculated. Results of the study showed that pin setting was critical for controlling the fill weight and the weight variation. The order of pin setting with pin 1 (closer to the powder chute) set to a relatively higher position and pin 4 (before ejection) set to a lower position was found to give higher fill weights with relatively lower weight variability. The powder bed height influenced the fill weight for poorly flowing powders. The capsule machine speed did not appear to significantly influence the fill weight. The fill weight and weight variation were found to depend on the flow property of the material. A large percentage of defective capsules was obtained using HPMC shell size #00. Some of the commonly observed defects included split caps and improperly closed filled capsules. In general, appropriate selection of pin settings and bed height can reduce the weight variability seen, especially with poorly flowing high-dose formulations. PMID- 15760055 TI - Heat and mass transfer scale-up issues during freeze drying: II. Control and characterization of the degree of supercooling. AB - This study aims to investigate the effect of the ice nucleation temperature on the primary drying process using an ice fog technique for temperature-controlled nucleation. In order to facilitate scale up of the freeze-drying process, this research seeks to find a correlation of the product resistance and the degree of supercooling with the specific surface area of the product. Freeze-drying experiments were performed using 5% wt/vol solutions of sucrose, dextran, hydroxyethyl starch (HES), and mannitol. Temperature-controlled nucleation was achieved using the ice fog technique where cold nitrogen gas was introduced into the chamber to form an "ice fog," thereby facilitating nucleation of samples at the temperature of interest. Manometric temperature measurement (MTM) was used during primary drying to evaluate the product resistance as a function of cake thickness. Specific surface areas (SSA) of the freeze-dried cakes were determined. The ice fog technique was refined to successfully control the ice nucleation temperature of solutions within 1 degrees C. A significant increase in product resistance was produced by a decrease in nucleation temperature. The SSA was found to increase with decreasing nucleation temperature, and the product resistance increased with increasing SSA. The ice fog technique can be refined into a viable method for nucleation temperature control. The SSA of the product correlates well with the degree of supercooling and with the resistance of the product to mass transfer (ie, flow of water vapor through the dry layer). Using this correlation and SSA measurements, one could predict scale-up drying differences and accordingly alter the freeze-drying process so as to bring about equivalence of product temperature history during lyophilization. PMID- 15760056 TI - Tablet formulation containing meloxicam and beta-cyclodextrin: mechanical characterization and bioavailability evaluation. AB - The purpose of this research was to evaluate beta-cyclodextrin (beta-CD) as a vehicle, either singly or in blends with lactose (spray-dried or monohydrate), for preparing a meloxicam tablet. Aqueous solubility of meloxicam in presence of beta-CD was investigated. The tablets were prepared by direct compression and wet granulation techniques. The powder blends and the granules were evaluated for angle of repose, bulk density, compressibility index, total porosity, and drug content. The tablets were subjected to thickness, diameter, weight variation test, drug content, hardness, friability, disintegration time, and in vitro dissolution studies. The effect of beta-CD on the bioavailability of meloxicam was also investigated in human volunteers using a balanced 2-way crossover study. Phase-solubility studies indicated an A(L)-type diagram with inclusion complex of 1:1 molar ratio. The powder blends and granules of all formulations showed satisfactory flow properties, compressibility, and drug content. All tablet formulations prepared by direct compression or wet granulation showed acceptable mechanical properties. The dissolution rate of meloxicam was significantly enhanced by inclusion of beta-CD in the formulations up to 30%. The mean pharmacokinetic parameters (C(max), K(e), and area under the curve [AUC](0 infinity)) were significantly increased in presence of beta-CD. These results suggest that beta-CD would facilitate the preparation of meloxicam tablets with acceptable mechanical properties using the direct compression technique as there is no important difference between tablets prepared by direct compression and those prepared by wet granulation. Also, beta-CD is particularly useful for improving the oral bioavailablity of meloxicam. PMID- 15760057 TI - The surface roughness of lactose particles can be modulated by wet-smoothing using a high-shear mixer. AB - The surface morphology of a-lactose monohydrate particles was modified by a new wet-smoothing process performed in a high-shear mixer using solvents. Successive steps of wetting and drying of lactose powders during rolling in the mixer's cylindrical bowl were performed. Smoothed particles were tested for size distribution, flow, and packing. The wet-smoothing process flattened the surface and rounded the edges of lactose particles. In comparison with original lactose, an improvement of powder packing and flow properties was evidenced. When the process was performed in the presence of a ternary agent such as magnesium stearate, the smoothing was improved. The evolution of rugosity during the smoothing process was assessed through a fractal descriptor of SEM picture. Atomic force microscopy and surface area measurements quantified the surface rugosity. A very significant reduction of the rugosity, more remarkable in the presence of magnesium stearate, was measured. This new process of powder wet smoothing allows the preparation of lactose particles with different degrees of smoothed surface for the control of flow and packing properties and particle particle interactions. PMID- 15760058 TI - Melt solidification technique: incorporation of higher wax content in ibuprofen beads. AB - The purpose of this study was to achieve incorporation of a higher amount of wax during the preparation of ibuprofen beads by a melt solidification technique for better integrity and prolonged drug release by using a combination of waxes. A mixture of cetyl alcohol (CA) and palmitic acid (PA) was used to improve the matrix integrity and drug release. The effect of variables such as CA, PA, and speed of agitation were studied using 3(3) factorial design. Yield, crushing strength, and drug release were analyzed using response surface methodology. The in vitro dissolution test did not show any significant improvement in the drug release. Scanning electron microscopy (SEM) showed that beads were spherical with a smooth surface, but after dissolution became rough and porous. Differential scanning calorimetry (DSC) studies showed that different solidification and erosion properties of waxes are responsible for the inability of waxes to retard drug release even at higher concentration. PMID- 15760059 TI - Molecular properties of ciprofloxacin-Indion 234 complexes. AB - The purpose of this research was to formulate tasteless complexes of ciprofloxacin with Indion 234 and to evaluate molecular properties of drug complexes. The effect of batch and column process, complexation time, temperature, and pH on ciprofloxacin loading on Indion 234 is reported. Drug resin complexes (DRC) were characterized by infrared spectroscopy, thermal analysis, and x-ray diffraction pattern. Ciprofloxacin release from DRC is obtained at salivary and gastric pH and in the presence of electrolytes. The efficient drug loading was evident in batch process using activated Indion 234 with a drug-resin ratio of 1:1.3. Drug complexation enhanced with pH from 1.2 to 6, while temperature did not affect the complexation process. Infrared spectroscopy revealed complexation of -NH (drug) with Indion 234. DRC are amorphous in nature. Drug release from DRC in salivary pH was insufficient to impart bitter taste. Volunteers rated the complex as tasteless and agreeable. Complete drug release was observed at gastric pH in 2 hours. The drug release was accelerated in the presence of electrolytes. Indion 234 is inexpensive, and the simple technique is effective for bitterness masking of ciprofloxacin. PMID- 15760060 TI - Effect of organogel components on in vitro nasal delivery of propranolol hydrochloride. AB - The purpose of this research was to evaluate in vitro transnasal sustained release ability of sorbitan monostearate (SMS) organogels in isopropyl myristate (IM). Organogels were prepared containing SMS (2.5%-20%) and water (5%-25%) in IM and analyzed microscopically for phase behavior. The effect of Tween surfactants on gel strength and in vitro nasal diffusion of propranolol is reported. The in vitro nasal release retardant effect of SMS and Tween 20 was investigated using factorial design. The microscopic changes in structure of organogel during in vitro nasal diffusion were studied. The water-holding capacity of SMS organogels in IM increased with SMS concentration. The release retardant effect with incorporation of cosurfactant was of the order of Tween 80 > Tween 60 > Tween 20. Gel strengthening and increased viscosity were evident with increased concentration of SMS and Tween 20. The 3-dimensional network of SMS molecules controls the diffusional drug release. The organogel system on nasal mucosa during diffusion is dynamic in nature and changes continuously with the time of diffusion. The water penetration in the organogel network results in percolation and emulsification of organogel, thus affecting the release. Organogels provided an effective barrier for diffusion of propranolol. The surface epithelium lining and the granular cellular structure of treated nasal mucosa were intact. PMID- 15760061 TI - Drug release properties of polyethylene-glycol-treated ciprofloxacin-Indion 234 complexes. AB - The polyethylene glycol (PEG) treatment of ciprofloxacin-Indion 234 complex was aimed to retard rapid ion exchange drug release at gastric pH. Ciprofloxacin loading on Indion 234 was performed in a batch process, and the amount of K(+) in Indion 234 displaced by drug with time was studied as equilibrium constant K(DM). Drug-resin complex (DRC) was treated with aqueous PEG solution (0.5%-2% wt/vol) of different molecular weights (MWs) for 2 to 30 minutes. The PEG-treated ciprofloxacin-Indion 234 complex was evaluated for particle size, water absorption time, and drug release at gastric pH. During drug loading on Indion 234, the equilibrium constant (K(DM)) increased rapidly up to 20 minutes with efficient drug loading. Increased time of immersion of the drug resinate in PEG solutions significantly retained higher size particles upon dehydration. The larger DRC particles showed longer water absorption times owing to compromised hydrating power. The untreated DRC showed insignificant drug release in deionized water; while at gastric pH, ciprofloxacin release was complete in 90 minutes. A trend of increased residual particle size, proportionate increase in water absorption time, and hence the retardation of release with time of immersion was evident in PEG-treated DRC. The time of immersion of DRC in PEG solution had predominant release retardant effect, while the effect of molecular weight of PEG was insignificant. Thus, PEG treatment of DRC successfully retards ciprofloxacin ion exchange release in acidic pH. PMID- 15760062 TI - Liposomal amikacin dry powder inhaler: effect of fines on in vitro performance. AB - The aim of the present investigation was to prepare and evaluate the influence of adding fines on the in vitro performance of liposomal amikacin dry powder inhaler (AMK LDPI) formulations. Liposomes composed of hydrogenated soyaphosphatidylcholine, cholesterol and saturated soyaphosphatidylglycerol (AMK 1), or stearylamine (AMK 2) were prepared by a reverse phase evaporation technique, extruded to reduce size and separated from unentrapped drug. Purified liposomal dispersion was subjected to lyophilization using optimized cryoprotectant to achieve maximum percentage drug retention (PDR). Lactose carrier in varying mass ratios with or without addition of fines in different mixing sequences was used to formulate AMK LDPI formulations. AMK LDPI formulations were characterized for angle of repose, compressibility index, dispersibility index, scanning electron microscopy, and fine particle fraction (FPF). PDR was found to be 97.6% +/- 2.2% for AMK1 and 98.5% +/- 1.9% for AMK2 using sucrose as optimized cryoprotectant in lipid:sucrose ratio of 1:4. Lactose carrier containing 10% fines (wt/wt) was found to be the optimum blend at 1:5 mass ratio of liposome:lactose. The addition of fines and the order of mixing of fines were found to influence the FPF with significantly different device fractions. FPF of AMK LDPI formulations using Rotahaler as the delivery device at 30, 60, and 90 L/min were found to be 21.85% +/- 2.2% and 24.6% +/- 2.4%, 25.9% +/- 1.8% and 29.2% +/- 2.1%, and 29.5% +/- 2.6% and 34.2% +/- 2.0% for AMK1 and AMK2, respectively. From the studies performed in this investigation, it was observed that liposomal charge, addition of fines and order of mixing fines, has a significant effect (P < .05) on in vitro deposition of drug from LDPI formulation. PMID- 15760063 TI - Response surface methodology for optimization and characterization of limonene based coenzyme Q10 self-nanoemulsified capsule dosage form. AB - The aim of this study was to systematically obtain a model of factors that would yield an optimized self-nanoemulsified capsule dosage form (SNCDF) of a highly lipophilic model compound, Coenzyme Q10 (CoQ). Independent variables such as amount of R-(+)-limonene (X1), surfactant (X2), and cosurfactant (X3), were optimized using a 3-factor, 3-level Box-Behnken statistical design. The dependent variables selected were cumulative percentage of drug released after 5 minutes (Y1) with constraints on drug release in 15 minutes (Y2), turbidity (Y3), particle size (Y4), and zeta potential (Y5). A mathematical relationship obtained, Y1 = 78.503 + 6.058X1 + 13.738X2 + 5.986X3 - 25.831X1(2) + 9.12X1X2 - 26.03 X1X3 - 38.67 X2(2) +11.02X2X3 - 15.55 X3(3) (r2 = 0.97), explained the main and quadratic effects, and the interaction of factors that affected the drug release. Response surface methodology (RSM) predicted the levels of factors X1, X2, and X3 (0.0344, 0.216, and 0.240, respectively), for a maximized response of Y1 with constraints of >90% release on Y2. The observed and predicted values of Y1 were in close agreement. In conclusion, the Box-Behnken experimental design allowed us to obtain SNCDF with rapid (>90%) drug release within 5 minutes with desirable properties of low turbidity and particle size. PMID- 15760064 TI - Evaluation of mucoadhesive properties of chitosan microspheres prepared by different methods. AB - The mucoadhesive properties of chitosan microspheres prepared by different methods were evaluated by studying the interaction between mucin and microspheres in aqueous solution. The interaction was determined by the measurement of mucin adsorbed on the microspheres. A strong interaction between chitosan microspheres and mucin was detected. The intensity of the interaction was dependent upon the method of preparation of chitosan microspheres and the amount of mucin added. The extent of mucus adsorption was proportional to the absolute values of the positive zeta potential of chitosan microspheres. The zeta potential in turn was found to be dependent upon the method of preparation of microspheres. The adsorption of type III mucin (1% sialic acid content) was interpreted using Freundlich or Langmuir adsorption isotherms. The values of r2 were greater for Langmuir isotherm as compared with Freundlich isotherm. The adsorption of a suspension of chitosan microspheres in the rat small intestine indicated that chitosan microspheres prepared by tripolyphosphate cross-linking and emulsification ionotropic gelation can be used as an excellent mucoadhesive delivery system. The microspheres prepared by glutaraldehyde and thermal cross linking showed good stability in HCl as compared with microspheres prepared by tripolyphosphate and emulsification ionotropic gelation. PMID- 15760065 TI - Nanoparticles containing ketoprofen and acrylic polymers prepared by an aerosol flow reactor method. AB - The purpose of this study was to outline the effects of interactions between a model drug and various acrylic polymers on the physical properties of nanoparticles prepared by an aerosol flow reactor method. The amount of model drug, ketoprofen, in the nanoparticles was varied, and the nanoparticles were analyzed for particle size distribution, particle morphology, thermal properties, IR spectroscopy, and drug release. The nanoparticles produced were spherical, amorphous, and had a matrix-type structure. Ketoprofen crystallization was observed when the amount of drug in Eudragit L nanoparticles was more than 33% (wt/wt). For Eudragit E and Eudragit RS nanoparticles, the drug acted as an effective plasticizer resulting in lowering of the glass transition of the polymer. Two factors affected the preparation of nanoparticles by the aerosol flow reactor method, namely, the solubility of the drug in the polymer matrix and the thermal properties of the resulting drug-polymer matrix. PMID- 15760066 TI - Examination of aqueous oxidized cellulose dispersions as a potential drug carrier. I. Preparation and characterization of oxidized cellulose phenylpropanolamine complexes. AB - Partially neutralized aqueous dispersions of oxidized cellulose (OC) (COOH content 24.2%; degree of neutralization [DN] 0.22-0.44; solid content 14.4% wt/wt), a biocompatible biodegradable polymer, were prepared and their use to entrap an amine drug was demonstrated. Phenylpropanolamine hydrochloride (PPA.HCl) was used as a model drug. OCA-PPA complexes were prepared by adding the drug solution to the OC dispersion. Light microscopy, powder x-ray diffractometry (PXRD), and Fourier-transform infrared (FT-IR) spectroscopy were used to characterize hydrated and dried OC and the OC-PPA complexes. Drug loading and drug-loading efficiency were calculated from high-performance liquid chromatography. Light microscopy revealed the partially neutralized OC to exist as swollen fibers in the dispersion. The degree of swelling increased with increasing DN of the OC. All dispersions, irrespective of DN, showed a pseudo plastic flow. The drug loading (12.6%-26.7%) and drug-loading efficiency (30% 48%) increased linearly with increasing DN and drug concentration. The PXRD of the OC-PPA complexes showed no diffraction peaks due to PPA, suggesting that the drug exists in the amorphous state. The FT-IR spectra of the complexes revealed the presence of an ionic linkage between OC and PPA. In conclusion, the results show that the aqueous OC dispersions can be used to molecularly entrap amine drugs to produce an OC-drug complex linked via an ionic linkage. PMID- 15760067 TI - Examination of aqueous oxidized cellulose dispersions as a potential drug carrier. II. In vitro and in vivo evaluation of phenylpropanolamine release from microparticles and pellets. AB - The purpose of this research is to investigate the release of phenylpropanolamine from oxidized cellulose-phenylpropanolamine (OC-PPA) complexes prepared using aqueous OC dispersions (degree of neutralization, DN, 0-0.44) and phenylpropanolamine-hydrochloride (PPA.HCl) (concentration, 0.5 M or 1.4 M) in vitro and in vivo. The results showed a faster drug release from the OC-PPA complex made using the OC dispersion with a DN value of 0.22 than from those prepared using dispersions with DN values of 0.29 to 0.44. No significant difference existed between the release profiles of OC-PPA microparticles made using OC dispersions with DN values of 0.29 to 0.44. OC-PPA complexes that contained smaller size particles or higher drug levels, or that were processed by freeze drying released PPA faster. Compared with microparticles, the pellets of OC-PPA complexes released PPA more slowly initially. An increase in pH or ionic strength of the dissolution medium increased the release of PPA, which is attributable to increased polymer hydration and solubilization at higher pH and ionic strength conditions. The OC-PPA pellets implanted subcutaneously in rats released 100% of their PPA in 9 to 12 hours. A good correlation was found between the in vivo and in vitro release data. Tissue pathology results showed no significant inflammatory tissue reactions. In conclusion, the partially ionized aqueous OC dispersions have the potential to be used as an implantable biodegradable carrier for amine drugs. PMID- 15760068 TI - Comparative in vitro and in vivo evaluation of matrix, osmotic matrix, and osmotic pump tablets for controlled delivery of diclofenac sodium. AB - The aim of this investigation was preparation and comparative evaluation of fabricated matrix (FM), osmotic matrix (OM), and osmotic pump (OP) tablets for controlled delivery of diclofenac sodium (DS). All formulations were evaluated for various physical parameters, and in vitro studies were performed on USP 24 dissolution apparatus II in pH 7.4 buffer and distilled water. In vivo studies were performed in 6 healthy human volunteers; the drug was assayed in plasma using HPLC, and results were compared with the performance of 2 commercial tablets of DS. Various pharmacokinetic parameters (ie, C(max), T(max), area under the curve [AUC(0-24)], and mean residence time) and relative bioavailability were compared. All fabricated formulations showed more prolonged and controlled DS release compared with commercial tablets studied. The OM and OP tablets, however, performed better than the matrix tablets. The rate and extent of drug release from FM1 matrix tablets (single polymer) was significantly different from that of FM2 (admixed polymers). Type of porosigenic agents and osmogens also influenced the drug release. Analysis of in vitro data by regression coefficient analysis revealed zero-order release kinetics for OM and OP tablets, while FM tablets exhibited Higuchi kinetics. In vivo results indicated prolonged blood levels with delayed peak and improved bioavailability for fabricated tablets compared to commercial tablets. It was concluded that the osmotic matrix and osmotic pump tablets could provide more prolonged, controlled, and gastrointestinal environmental-independent DS release that may result in an improved therapeutic efficacy and patient compliance. PMID- 15760069 TI - Preparation and evaluation of a novel buccal adhesive system. AB - The aim of the present study was to prepare and evaluate a novel buccal adhesive system (NBAS) containing propranolol hydrochloride (PH). A special punch was fabricated and used while preparing an NBAS. Solubility of PH in phosphate buffer solution (pH 6.6), partition coefficient between phosphate buffer (pH 6.6) and 1 octanol, and permeability coefficient through the porcine buccal mucosa were performed and found to be 74.66 mg/mL, 5.17, and 5.6, respectively. Stability of NBAS was determined in natural human saliva, and it was found that both PH and device are stable in human saliva. NBAS was evaluated by weight uniformity, thickness, hardness, friability, swelling, mucoadhesive strength, in vitro drug release, and in vivo human acceptability studies. Swelling index was higher (4.4) for formulations containing hydroxyl propyl methyl cellulose (HPMC) K4M alone, and it decreases with its decreasing concentration in the NBAS. Mucoadhesive strength (MS) was measured by using a modified apparatus. All NBASs showed higher MS with porcine buccal mucosa when compared with that of rabbit buccal mucosa. NBASs containing carbopol (CP) 934P and HPMC K4M at the ratio of 1:1 showed higher MS (44.76 g) with porcine buccal mucosa when compared with 1:2 (39.76 g), 0:1 (23.29 g), and 1:0 (22.22 g) ratios, respectively. The mechanism of PH release was found to be by non-Fickian diffusion (value of "n" between 0.5 and 1.0) and followed first order kinetics. In vivo human acceptability study showed that the newly prepared NBAS was comfortable in the human buccal cavity. It can be concluded that NBAS is a superior, novel system that overcomes the drawback associated with the conventional buccal adhesive tablet. PMID- 15760070 TI - Formulation design and optimization of mouth dissolve tablets of nimesulide using vacuum drying technique. AB - The purpose of this research was to develop mouth dissolve tablets of nimesulide. Granules containing nimesulide, camphor, crospovidone, and lactose were prepared by wet granulation technique. Camphor was sublimed from the dried granules by exposure to vacuum. The porous granules were then compressed. Alternatively, tablets were first prepared and later exposed to vacuum. The tablets were evaluated for percentage friability, wetting time, and disintegration time. In the investigation, a 32 full factorial design was used to investigate the joint influence of 2 formulation variables: amount of camphor and crospovidone. The results of multiple linear regression analysis revealed that for obtaining a rapidly disintegrating dosage form, tablets should be prepared using an optimum concentration of camphor and a higher percentage of crospovidone. A contour plot is also presented to graphically represent the effect of the independent variables on the disintegration time and percentage friability. A checkpoint batch was also prepared to prove the validity of the evolved mathematical model. Sublimation of camphor from tablets resulted in superior tablets as compared with the tablets prepared from granules that were exposed to vacuum. The systematic formulation approach helped in understanding the effect of formulation processing variables. PMID- 15760071 TI - Effect of starting material particle size on its agglomeration behavior in high shear wet granulation. AB - The effect of anhydrous lactose particle size distribution on its performance in the wet granulation process was evaluated. Three grades of anhydrous lactose were used in the study: "as is" manufacturer grade and 2 particle size fractions obtained by screening of the 60M lactose. Particle growth behavior of the 3 lactose grades was evaluated in a high shear mixer. Compactibility and porosity of the resulting granules were also evaluated. A uniaxial compression test on moist agglomerates of the 3 lactose grades was performed in an attempt to explain the mechanism of particle size effect observed in the high shear mixer. Particle growth of anhydrous lactose in the high shear mixer was inversely related to the particle size of the starting material. In addition, granulation manufactured using the grade with the smallest particle size was more porous and demonstrated enhanced compactibility compared with the other grades. Compacts with similar porosity and low liquid saturation demonstrated brittle behavior and their breakage strength was inversely related to lactose particle size in the uniaxial compression test, suggesting that material with smaller particle size may exhibit more pronounced nucleation behavior during wet granulation. On the other hand, compacts prepared at higher liquid saturation and similar compression force exhibited more plastic behavior and showed lower yield stress for the grade with smallest particle size. The lower yield stress of compacts prepared with this grade may indicate a higher coalescence tendency for its granules during wet granulation. PMID- 15760072 TI - In vitro evaluation of a morphine polymeric complex: flowability behavior and dissolution study. AB - The purpose of this research was to perform a granulometrical and flow properties study of a morphine polymeric complex and determine the influence of 3 variables- particle size of complex, pH value, and ionic strength of the dissolution medium- on the dissolution behavior. The morphine-Eudragit L complex was produced in aqueous medium from morphine hydrochloride saturated solution and Eudragit L 30D diluted until 12% wt/vol and partially neutralized (40%). To determine the rheological behavior of the complex, several rheological tests were developed: bulk and tapped densities, Hausner ratio, angle of repose, and flow rate. The results corresponding to the technological study suggest that the 100- to 250 microm fraction can be considered as free flowing powder. In relation to the dissolution behavior of the complex, the results indicate that the ionic strength has been detected as the most influencing factor when values below physiological conditions are used. In conclusion, no technological problems for the production of further solid dosage forms are expected. Furthermore, no changes in the dissolution profiles of the complex have been detected when ionic strength values are inside the physiological range. PMID- 15760073 TI - A method to determine the incorporation capacity of camptothecin in liposomes. AB - The purpose of this study was to establish a new experimental approach to determine the maximum amount of camptothecin (CPT) that can be incorporated in liposomes, and to use this method to compare the CPT-incorporation capacity of various liposome formulations. Small, CPT-saturated liposomes were prepared by dispersing freeze-dried blends of lipids and drug in phosphate buffer, and subsequent probe-sonication. Excess precipitated CPT could be separated from the liposomes by ultracentrifugation. The small and homogeneous liposome size obtained gave a good and reproducible recovery of liposomes in the supernatant (>80%), whereas the acidic pH (pH 6.0) kept CPT in its hydrophobic lactone form, which is poorly soluble in the buffer. The maximum CPT-incorporation capacity of 12 different liposome formulations was investigated, using the described method, and was found to vary widely. With liposomes made of neutral and anionic phospholipids, the solubility of CPT in the buffer was improved by approximately a factor of 10 (from 2.7 to 15-50 microg/mL) as compared with buffer. With cationic liposomes containing 1,2-dioleoyl-3-trimethyl-ammonium-propane (DOTAP), a maximum CPT-solubilization of 100-fold, the buffer solubility was reached, probably owing to an electrostatic interaction between the cationic lipids and the carboxylate-CPT isomer. Increasing DOTAP fractions within egg phosphatidylcholine (EPC)/DOTAP liposomes reached a CPT-incorporation plateau at 20 mol% DOTAP. The presented approach appears suitable to study the incorporation capacity of any drug component within small vesicles as long as the liposome incorporation is high relative to the intrinsic water solubility of the drug. PMID- 15760074 TI - Effect of molecular weight of hydrolyzed gelatin on its binding properties in tablets: a technical note. PMID- 15760075 TI - Effect of formulation variables on preparation and evaluation of gelled self emulsifying drug delivery system (SEDDS) of ketoprofen. AB - The purpose of this study was to formulate a gelled self-emulsifying drug delivery system (SEDDS) containing ketoprofen as an intermediate in the development of sustained release solid dosage form. Captex 200 (an oil), Tween 80 (a surfactant), and Capmul MCM (a cosurfactant) were used to formulate SEDDS. Silicon dioxide was used as a gelling agent, which may aid in solidification and retardation of drug release. Effect of concentrations of cosurfactant and gelling agent on emulsification process and in vitro drug diffusion was studied using 3(2) factorial design. Multiple regression analysis data and response surfaces obtained showed that liquid crystal phase viscosity increased significantly with increasing amount of silicon dioxide, which in turn caused an increase in average droplet size of resultant emulsion and slower drug diffusion. Drug release from the formulation increased with increasing amount of cosurfactant. PMID- 15760076 TI - Preparation and evaluation of diltiazem hydrochloride-Gelucire 43/01 floating granules prepared by melt granulation. AB - The basic objective of this study was to explore the application of Gelucire 43/01 for the design of multi-unit floating systems of a highly water-soluble drug diltiazem HCl. Diltiazem HCl-Gelucire 43/01 granules were prepared by melt granulation technique. The granules were evaluated for in vitro and in vivo floating ability, surface topography, and in vitro drug release. Aging effect on storage was evaluated using scanning electron microscopy, hot stage polarizing microscopy (HSPM), differential scanning calorimetry (DSC), and in vitro drug release. Granules were retained in stomach at least for 6 hours. Approximately 65% to 80% drug was released over 6 hours with initial fast release from the surface. Surface topography, HSPM, DSC study of the aged samples showed phase transformation of Gelucire. The phase transformation also caused significant increase in drug release. In conclusion, hydrophobic lipid, Gelucire 43/01, can be considered as an effective carrier for design of a multi-unit floating drug delivery system of highly water-soluble drugs such as diltiazem HCl. PMID- 15760077 TI - Crystallo-co-agglomeration: a novel technique to obtain ibuprofen-paracetamol agglomerates. AB - The purpose of this research was to obtain directly compressible agglomerates of ibuprofen-paracetamol containing a desired ratio of drugs using a crystallo-co agglomeration technique. Crystallo-co-agglomeration is an extension of the spherical crystallization technique, which enables simultaneous crystallization and agglomeration of 2 or more drugs or crystallization of a drug and its simultaneous agglomeration with another drug or excipient. Dichloromethane (DCM) water system containing polyethylene glycol (PEG) 6000, polyvinyl pyrollidone, and ethylcellulose was used as the crystallization system. DCM acted as a good solvent for ibuprofen and bridging liquid for agglomeration. The process was performed at pH 5, considering the low solubility of ibuprofen and the stability of paracetamol. Loss of paracetamol was reduced by maintaining a low process temperature and by the addition of dextrose as a solubility suppressant. The agglomerates were characterized by differential scanning calorimetry, powder x ray diffraction (PXRD), and scanning electron microscopy and were evaluated for tableting properties. The spherical agglomerates contained an ibuprofen paracetamol ratio in the range of 1.23 to 1.36. Micromeritic, mechanical, and compressional properties of the agglomerates were affected by incorporated polymer. The PXRD data showed reduction in intensities owing to dilution and reduced crystallinity. Thermal data showed interaction between components at higher temperature. Ethylcellulose imparted mechanical strength to the agglomerates as well as compacts. The agglomerates containing PEG have better compressibility but drug release in the initial stages was affected owing to asperity melting, yielding harder compacts. The agglomeration and properties of agglomerates were influenced by the nature of polymer. PMID- 15760078 TI - Effect of pH-sodium lauryl sulfate combination on solubilization of PG-300995 (an anti-HIV agent): a technical note. PMID- 15760080 TI - Measurement of fluid viscosity at microliter volumes using quartz impedance analysis. AB - The purpose of this work was to measure viscosity of fluids at low microliter volumes by means of quartz crystal impedance analysis. To achieve this, a novel setup was designed that allowed for measurement of viscosity at volumes of 8 to 10 microL. The technique was based on the principle of electromechanical coupling of piezoelectric quartz crystals. The arrangement was simple with measurement times ranging from 2 to 3 minutes. The crystal setup assembly did not impose any unwanted initial stress on the unloaded quartz crystal. Quartz crystals of 5- and 10-MHz fundamental frequency were calibrated with glycerol-water mixtures of known density and viscosity prior to viscosity measurements. True frequency shifts, for the purpose of this work, were determined followed by viscosity measurement of aqueous solutions of sucrose, urea, PEG-400, glucose, and ethylene glycol at 25 degrees C +/- 0.5 degrees C. The measured viscosities were found to be reproducible and consistent with the values reported in the literature. Minor inconsistencies in the measured resistance and frequency shifts did not affect the results significantly, and were found to be experimental in origin rather than due to electrode surface roughness. Besides, as expected for a viscoelastic fluid, PEG 8000 solutions, the calculated viscosities were found to be less than the reported values due to frequency dependence of storage and loss modulus components of complex viscosity. From the results, it can be concluded that the present setup can provide accurate assessment of viscosity of Newtonian fluids and also shows potential for analyzing non-Newtonian fluids at low microliter volumes. PMID- 15760081 TI - Transdermal delivery of zidovudine (AZT): the effects of vehicles, enhancers, and polymer membranes on permeation across cadaver pig skin. AB - The purpose of this study was to investigate the effects of vehicles, enhancers, and polymer membranes on 3'-azido-3'-deoxythymidine (AZT) permeation across cadaver pig skin. Four binary vehicles (ethanol/water, isopropyl alcohol/water, polyethylene glycol 400/water, and ethanol/isopropyl myristate [IPM]) were tested for AZT solubility and permeability across pig skin; ethanol/IPM (50/50, vol/vol) demonstrated the highest AZT flux (185.23 microg/cm2/h). Next, the addition of various concentrations of different enhancers (N-methyl-2-pyrrolidone [NMP], oleic acid, and lauric acid) to different volume ratios of ethanol/IPM was investigated for their effect on AZT solubility and permeability across pig skin. The use of 2 combinations (ethanol/IPM [20/80] plus 10% NMP and ethanol/IPM [30/70] plus 10% NMP) resulted in increased AZT solubility (42.6 and 56.27 mg/mL, respectively) and also high AZT flux values (284.92 and 460.34 microg/cm2/h, respectively) without appreciable changes in lag times (6.25 and 7.49 hours, respectively) when compared with formulations using only ethanol/IPM at 20/80 and 30/70 volume ratios without addition of the enhancer NMP. Finally, AZT permeation across pig skin covered with a microporous polyethylene (PE) membrane was investigated. The addition of the PE membrane to the pig skin reduced AZT flux values to 50% of that seen with pig skin alone. However, the AZT flux value attained with ethanol/IPM (30/70) plus 10% NMP was 215.31 microg/cm2/h, which was greater than the target flux (208 mug/cm2/h) needed to maintain the steady-state plasma concentration in humans. The results obtained from this study will be helpful in the development of an AZT transdermal drug delivery system. PMID- 15760082 TI - In vivo release kinetics of octreotide acetate from experimental polymeric microsphere formulations using oil/water and oil/oil processes. AB - The purpose of the present study was to characterize the in vivo release kinetics of octreotide acetate from microsphere formulations designed to minimize peptide acylation and improve drug stability. Microspheres were prepared by a conventional oil/water (o/w) method or an experimental oil/oil (o/o) dispersion technique. The dosage forms were administered subcutaneously to a rat animal model, and serum samples were analyzed by radioimmunoassay over a 2-month period. An averaged kinetic profile from each treatment group, as a result, was treated with fractional differential equations. The results indicated that poly(l lactide) microspheres prepared by the o/o dispersion technique provided lower area under the curve (AUC) values during the initial diffusion-controlled release phase, 7.79 ngxd/mL, versus 75.8 ngxd/mL for the o/w batch. During the subsequent erosion-controlled release phase, on the other hand, the o/o technique yielded higher AUC values, 123 ngxd/mL, versus 42.2 ngxd/mL for the o/w batch. The differences observed between the 2 techniques were attributed to the site of drug incorporation during the manufacturing process, given that microspheres contain both porous hydrophilic channels and dense hydrophobic matrix regions. An o/o dispersion technique was therefore expected to produce microspheres with lower incorporation in the aqueous channels, which are responsible for diffusion mediated drug release. PMID- 15760083 TI - Effect of potassium chloride and cationic drug on swelling, erosion and release from kappa-carrageenan matrices. AB - The basic objective of this work was to study the effect of model cationic drug metformin HCl on swelling and erosion and, in turn, the release of KCl and drug itself, from the kappa-carrageenan matrices. Water uptake by the matrix up to 2 hours was found to increase with KCl concentration from the plain matrix. Erosion was not affected by concentration of KCl. Incorporation of drug favors water uptake, but in presence of KCl it was found to be reduced. Drug-containing matrices have shown higher release of KCl as compared with plain batches. Drug release was retarded as KCl concentration increased up to 5%, above which the reduced cohesivity of the matrix caused increase in drug release. PMID- 15760084 TI - Effects of water vapor absorption on the physical and chemical stability of amorphous sodium indomethacin. AB - This study reports on the effects that water absorbed into amorphous sodium indomethacin (NaIMC) can have on simultaneous tendencies to crystallize to its trihydrate form and to undergo base-catalyzed hydrolysis because of the plasticizing effects of water on molecular mobility. Measurement of water vapor absorption at 30 degrees C and powder x-ray diffraction patterns as a function of relative humidity (RH) reveal that upon exposure to 21% RH, NaIMC does not crystallize over a 2-month period. Measurements of the glass transition temperature as a function of such exposure reveals a change in T(g) from 121 degrees C, dry, to 53 degrees C at 21% RH, such that T(g) at 21% RH is approximately 13 degrees C above the highest storage temperature of 40 degrees C used in the study. At 56% RH and higher, however, crystallization to the trihydrate occurs rapidly; although over the 2-month period, crystallization was never complete. Assessment of chemical degradation by high-performance liquid chromatography analysis revealed significant instability at 21% RH; whereas at higher RH, the extent of chemical degradation was reduced, reflecting the greater crystallization to the more chemically stable crystalline form. It is concluded that when amorphous forms of salts occur in solid dosage forms, the simultaneous effects of enhanced water vapor sorption on crystallization and chemical degradation must be considered, particularly when assessing solid-state chemical degradation at higher temperatures and RH (eg, 40 degrees C 75% RH). PMID- 15760085 TI - Preparation and in vivo toxicity study of solid lipid microparticles as carrier for pulmonary administration. AB - The purpose of this research was to investigate the effects of processing conditions on the characteristics of solid lipid microparticles (SLM) with a potential application as carriers for pulmonary administration. Compritol (5.0% wt/wt) SLM dispersions were prepared by rotor-stator homogenization, at different surfactant concentrations and emulsification times. The SLM were characterized, in terms of morphology and size, after lyophilization and sterilization by autoclaving process. In vivo assessment was carried out in rats by intratracheal instillation of either placebo or SLM dispersion, and by bronchoalveolar lavage for cytological analysis. Mean particle size of 4 to 5 microm was achieved using 0.3% and 0.4% (wt/wt) of emulsifier (Poloxamer 188) and emulsification times of 2 and 5 minutes. The particles showed spherical shape and smooth surface. The morphology of microparticles, the size, and the size distribution were not substantially modified after lyophilization and sterilization. Total cell counts showed no significant differences between placebo and SLM 0.5% or 2.5% groups. Regarding cytology, percentage of polymorphonuclear neutrophils and macrophages did not significantly differ between groups. These results suggest that a single intratracheal administration of the SLMs does not induce a significant inflammatory airway response in rats and that the SLMs might be a potential carrier for encapsulated drug via the pulmonary route. PMID- 15760086 TI - Crystal structure determination of thymoquinone by high-resolution X-ray powder diffraction. AB - The crystal structure of 2-isopropyl-5-methyl-1,4-benzoquinone (thymoquinone) and its thermal behavior--as necessary physical and chemical properties--were determined in order to enhance the current understanding of thymoquinone chemical action by using high resolution x-ray powder diffraction, Fourier transform infrared spectroscopy (FTIR), and 3 thermo-analytical techniques thermogravimetric analysis (TGA), differential thermal analysis (DTA), and differential scanning calorimetry (DSC). The findings obtained with high resolution x-ray powder diffraction and molecular location methods based on a simulated annealing algorithm after Rietveld refinement showed that the triclinic unit cell was a = 6.73728(8) A, b = 6.91560(8) A, c = 10.4988(2) A, alpha = 88.864(2) degrees, beta = 82.449(1) degrees, gamma = 77.0299(9) degrees; cell volume = 472.52(1) A3, Z = 2, and space group P1. In addition, FTIR spectrum revealed absorption bands corresponding to the carbonyl and C-H stretching of aliphatic and vinylic groups characteristically observed in such p-benzoquinones. Also, a chemical decomposition process starting at 65 degrees C and ending at 213 degrees C was noted when TGA was used. DSC allowed for the determination of onset at 43.55 degrees C and a melting enthalpy value of DeltaH(m) = 110.6 J/g. The low value obtained for the fusion point displayed a van der Waals pattern for molecular binding, and the thermograms performed evidence that thymoquinone can only be found in crystalline triclinic form, as determined by DRX methods. PMID- 15760087 TI - A novel approach in the assessment of polymeric film formation and film adhesion on different pharmaceutical solid substrates. AB - The purpose of this study was to evaluate the nature of film formation on tablets with different compositions, using confocal laser scanning microscopy (CLSM), and to measure film adhesion via the application of a novel "magnet probe test." Three excipients, microcrystalline cellulose (MCC), spray-dried lactose monohydrate, and dibasic calcium phosphate dihydrate, were individually blended with 0.5% magnesium stearate, as a lubricant, and 2.5% tetracycline HCl, as a fluorescent marker, and were compressed using a Carver press. Tablets were coated with a solution consisting of 7% hydroxypropyl methylcellulose (HPMC) phthalate (HP-55), and 0.5% cetyl alcohol in acetone and isopropanol (11:9). The nature of polymer interaction with the tablets and coating was evaluated using CLSM and a designed magnet probe test. CLSM images clearly showed coating efficiency, thickness, and uniformity of film formation, and the extent of drug migration into the film at the coating interfaces of tablets. Among the excipients, MCC demonstrated the best interface for both film formation and uniformity in thickness relative to lactose monohydrate and dibasic calcium phosphate dihydrate. The detachment force of the coating layers from the tablet surfaces, as measured with the developed magnet probe test, was in the order of MCC>lactose monohydrate>dibasic calcium phosphate dihydrate. It was also shown that the designed magnet probe test provides reliable and reproducible results when used for measurement of film adhesion and bonding strength. PMID- 15760088 TI - Spherical composite particles of rice starch and microcrystalline cellulose: a new coprocessed excipient for direct compression. AB - Composite particles of rice starch (RS) and microcrystalline cellulose were fabricated by spray-drying technique to be used as a directly compressible excipient. Two size fractions of microcrystalline cellulose, sieved (MCS) and jet milled (MCJ), having volumetric mean diameter (D50) of 13.61 and 40.51 microm, respectively, were used to form composite particles with RS in various mixing ratios. The composite particles produced were evaluated for their powder and compression properties. Although an increase in the microcrystalline cellulose proportion imparted greater compressibility of the composite particles, the shape of the particles was typically less spherical with rougher surface resulting in a decrease in the degree of flowability. Compressibility of composite particles made from different size fractions of microcrystalline cellulose was not different; however, using MCJ, which had a particle size range close to the size of RS (D50 = 13.57 microm), provided more spherical particles than using MCS. Spherical composite particles between RS and MCJ in the ratio of 7:3 (RS-MCJ-73) were then evaluated for powder properties and compressibility in comparison with some marketed directly compressible diluents. Compressibility of RS-MCJ-73 was greater than commercial spray-dried RS (Eratab), coprocessed lactose and microcrystalline cellulose (Cellactose), and agglomerated lactose (Tablettose), but, as expected, lower than microcrystalline cellulose (Vivapur 101). Flowability index of RS-MCJ-73 appeared to be slightly lower than Eratab but higher than Vivapur 101, Cellactose, and Tablettose. Tablets of RS-MCJ-73 exhibited low friability and good self-disintegrating property. It was concluded that these developed composite particles could be introduced as a new coprocessed direct compression excipient. PMID- 15760089 TI - Computational fluid dynamics modeling of the paddle dissolution apparatus: agitation rate, mixing patterns, and fluid velocities. AB - The purpose of this research was to further investigate the hydrodynamics of the United States Pharmacopeia (USP) paddle dissolution apparatus using a previously generated computational fluid dynamics (CFD) model. The influence of paddle rotational speed on the hydrodynamics in the dissolution vessel was simulated. The maximum velocity magnitude for axial and tangential velocities at different locations in the vessel was found to increase linearly with the paddle rotational speed. Path-lines of fluid mixing, which were examined from a central region at the base of the vessel, did not reveal a region of poor mixing between the upper cylindrical and lower hemispherical volumes, as previously speculated. Considerable differences in the resulting flow patterns were observed for paddle rotational speeds between 25 and 150 rpm. The approximate time required to achieve complete mixing varied between 2 to 5 seconds at 150 rpm and 40 to 60 seconds at 25 rpm, although complete mixing was achievable for each speed examined. An analysis of CFD-generated velocities above the top surface of a cylindrical compact positioned at the base of the vessel, below the center of the rotating paddle, revealed that the fluid in this region was undergoing solid body rotation. An examination of the velocity boundary layers adjacent to the curved surface of the compact revealed large peaks in the shear rates for a region within ~3 mm from the base of the compact, consistent with a "grooving" effect, which had been previously seen on the surface of compacts following dissolution, associated with a higher dissolution rate in this region. PMID- 15760090 TI - Anti-Helicobacter pylori effect of mucoadhesive nanoparticles bearing amoxicillin in experimental gerbils model. AB - The purpose of the present study was to design mucoadhesive gliadin nanoparticles (GNP) containing amoxicillin and to evaluate their effectiveness in eradicating Helicobacter pylori. GNP-bearing amoxicillin (AGNP) was prepared by desolvation method. The effect of process variables such as gliadin concentration and initial drug loading on particle size, shape, percent payload, percent entrapment efficiency, in vitro release profile, and mucoadhesive property of GNP was assessed. Rhodamine isothiocyanate-entrapped GNP formulations were prepared to evaluate their in vivo gastric mucoadhesive property in albino rats. With increasing gliadin concentration, the mucoadhesive property of GNP increased. Typically, the maximum amount of nanoparticles remaining was 82 +/- 4%, which represented a stronger mucoadhesive propensity and specificity of GNP toward the stomach. In vitro antimicrobial activity of AGNP was evaluated by growth inhibition studies on an isolated H pylori strain. The time required for complete eradication was higher in AGNP than in amoxicillin because of the controlled drug delivery of amoxicillin from AGNP. In vivo clearance of H pylori following oral administration of AGNP to infected Mongolian gerbils was examined. Amoxicillin and AGNP both showed anti-H pylori effects in this experimental model of infection, but the required dose for complete eradication was less in AGNP than in amoxicillin. In conclusion, AGNP eradicated H pylori from the gastrointestinal tract more effectively than amoxicillin because of the prolonged gastrointestinal residence time attributed to mucoadhesion. A dosage form containing mucoadhesive nanoparticles bearing a potential antibiotic should be useful for the complete eradication of H pylori. PMID- 15760091 TI - The influence of the copolymer composition on the diltiazem hydrochloride release from a series of pH-sensitive poly[(N-isopropylacrylamide)-co-(methacrylic acid)] hydrogels. AB - A series of poly[(N-isopropylacrylamide)-co-(methacrylic acid)] (P[(N-iPAAm)-co (MAA)]) hydrogels was investigated to determine the composition that exhibits a better pH-modulated release of diltiazem hydrochloride (DIL.HCl). For this purpose hydrogel slabs were loaded with DIL.HCl by the immersion method, and its release under acidic medium (0.1N HCl, pH 1.2) and in phosphate buffer pH 7.2, using United States Pharmacopeia (USP) 24 Apparatus 1, was investigated. According to the results from the slabs, copolymers with 85% mol N-iPAAm content were selected to prepare tablets with different particle size. The effect of pH and particle size changes on DIL.HCl release from these last hydrogel tablets was investigated by a stepwise pH variation of the dissolution medium. The amount of DIL.HCl released from high N-iPAAm content copolymer slabs under acidic pH medium was not only very low but it was also released at a slow rate. In the 85% N-iPAAm tablets, significant differences between and within release profiles were found as a function of particle size and pH, respectively. A relationship between particle size and release rate has been found. The lower DIL.HCl release at acidic pH from enriched N-iPAAm copolymers is interpreted by a cooperative thermal- and pH-collapse. Although for the whole range of copolymer composition a dependence of the equilibrium of swelling on the pH was found, DIL.HCl release experiments indicated that hydrogels with 85% mol N-iPAAm are the more adequate to be used for modulated drug delivery systems. Additionally, the particle size of the tablet can be used to tailor the release rate. PMID- 15760092 TI - Gastroretentive drug delivery system of ranitidine hydrochloride: formulation and in vitro evaluation. AB - The purpose of this research was to prepare a gastroretentive drug delivery system of ranitidine hydrochloride. Guar gum, xanthan gum, and hydroxypropyl methylcellulose were evaluated for gel-forming properties. Sodium bicarbonate was incorporated as a gas-generating agent. The effects of citric acid and stearic acid on drug release profile and floating properties were investigated. The addition of stearic acid reduces the drug dissolution due to its hydrophobic nature. A 3(2) full factorial design was applied to systemically optimize the drug release profile. The amounts of citric acid anhydrous (X1) and stearic acid (X2) were selected as independent variables. The times required for 50% (t50) and 80% drug dissolution (t80), and the similarity factor f2 were selected as dependent variables. The results of the full factorial design indicated that a low amount of citric acid and a high amount of stearic acid favors sustained release of ranitidine hydrochloride from a gastroretentive formulation. A theoretical dissolution profile was generated using pharmacokinetic parameters of ranitidine hydrochloride. The similarity factor f2 was applied between the factorial design batches and the theoretical dissolution profile. No significant difference was observed between the desired release profile and batches F2, F3, F6, and F9. Batch F9 showed the highest f2 (f2 = 75) among all the batches, and this similarity is also reflected in t50 (approximately 214 minutes) and t80 (approximately 537 minutes) values. These studies indicate that the proper balance between a release rate enhancer and a release rate retardant can produce a drug dissolution profile similar to a theoretical dissolution profile. PMID- 15760093 TI - Exposure-response relationships and drug interactions of sirolimus. AB - Sirolimus (rapamycin, RAPAMUNE, RAPA) is an immunosuppressive agent used for the prophylaxis of renal allograft rejection and exhibits an immunosuppressive mechanism that is distinct from that for cyclosporine and tacrolimus. The purpose of this manuscript is to discuss the exposure-response relationships and drug interactions of sirolimus. The various factors affecting sirolimus whole blood exposure included first-pass extraction, formulation, food, demographics, liver disease, assay method, and interacting drugs. Clinically significant effects caused by food, pediatric age, hepatic impairment, and interacting drugs require recommendations for the safe and efficacious use of sirolimus in renal allograft patients. An exposure-response model based on multivariate logistic regression was developed using the interstudy data from 1832 renal allograft patients. The analysis revealed an increased probability of acute rejection for sirolimus troughs <5 ng/mL, cyclosporine troughs <150 ng/mL, human leukocyte antigen (HLA) mismatches > or =4, and females. The outcomes suggested that individualization of sirolimus doses immediately after transplantation, based on HLA mismatch and sex, would likely decrease the probability of acute rejections in renal allograft recipients who receive concomitant sirolimus, cyclosporine (full-dose), and corticosteroid therapy. Sirolimus is a substrate for both Cytochrome P450 3A (CYP3A) and P-glycoprotein (P-gp) and undergoes extensive first-pass extraction. Drugs that are known to inhibit or induce these proteins may potentially affect sirolimus whole blood exposure. In healthy volunteers, cyclosporine, diltiazem, erythromycin, ketoconazole, and verapamil significantly increased sirolimus whole blood exposure, and rifampin significantly decreased sirolimus exposure. However, sirolimus whole blood exposure was not affected by acyclovir, atorvastatin, digoxin, ethinyl estradiol/norgestrel, glyburide, nifedipine, or tacrolimus. Among the 15 drugs studied, sirolimus significantly increased the exposures of only erythromycin and S-(-)verapamil. PMID- 15760094 TI - Anionic liposomal delivery system for DNA transfection. AB - The present study investigates the use of novel anionic lipoplexes composed of physiological components for plasmid DNA delivery into mammalian cells in vitro. Liposomes were prepared from mixtures of endogenously occurring anionic and zwitterionic lipids, 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (DOPG) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), respectively, at a molar ratio of 17:83 (DOPG:DOPE). Anionic lipoplexes were formed by complexation between anionic liposomes and plasmid DNA molecules encoding green fluorescence protein (GFP) using Ca2+ ions. Transfection and toxicity were evaluated in CHO-K1 cells using flow cytometry and propidium iodide staining, respectively. Controls included Ca2+-DNA complexes (without lipids), anionic liposomes (no Ca2+), and a cationic liposomal formulation. Efficient delivery of plasmid DNA and subsequent GFP expression was achieved using anionic lipoplexes. Transfection efficiency increased with Ca2+ concentration up to 14 mM Ca2+, where transfection efficiency was 7-fold higher than in untreated cells, with minimum toxicity. Further increase in Ca2+ decreased transfection. Transfection efficiency of anionic lipoplexes was similar to that of cationic liposomes (lipofectAmine), whereas their toxicity was significantly lower. Ca2+ DNA complexes exhibited minimal and irregular transfection with relatively high cytotoxicity. A model was developed to explain the basis of anionic lipoplex uptake and transfection efficacy. Effective transfection is explained on the formation of nonbilayer hexagonal lipid phases. Efficient and relatively safe DNA transfection using anionic lipoplexes makes them an appealing alternative to be explored for gene delivery. PMID- 15760095 TI - The role of halogen substitution in classical cannabinoids: a CB1 pharmacophore model. AB - The presence of halogens within the classical cannabinoid structure leads to large variations in the compounds' potencies and affinities for the CB1 receptors. To explore the structure activity relationships within this class of analogs we have used a series of halogen-substituted (-)-Delta8 tetrahydrocannabinol analogs and compared their affinities for the CB1 cannabinoid receptor. Our results indicate that halogen substitution at the end carbon of the side chain leads to an enhancement in affinity with the bulkier halogens (Br, I) producing the largest effects. Conversely, 2-iodo substitution on the phenolic ring leads to a 2-fold reduction in affinity while iodo substitution in the C1'-position of the side chain lowers the compound's affinity for CB1 by more than 8-fold. The pharmacophoric requirements resulting from halogen-substitution are explored using computer modeling methods. PMID- 15760096 TI - Regional permeability of salmon calcitonin in isolated rat gastrointestinal tracts: transport mechanism using Caco-2 cell monolayer. AB - The objective of the study was to determine the region of maximum permeation of salmon calcitonin (sCT) through the gastrointestinal tract and to investigate the mechanism of permeation. For regional permeability determination, male Sprague Dawley rats (250-300 g) were anesthetized and the gastrointestinal tissues were isolated. Stomach, duodenum, jejunum, ileum, or colon tissues were mounted on Navicyte side-by-side diffusion apparatus. Salmon calcitonin solutions (50 microm in phosphate-buffered saline, pH 7.4, 37 degrees C) were added to the donor side, and the samples were removed from the receiver compartment and analyzed by competitive radioimmunoassay (RIA). For mechanistic studies, Caco-2 cells were grown on Transwell inserts (0.4-microm pore size, 0.33 cm2 area) in a humidified 37 degrees C incubator (with 5% CO2). Transport experiments were conducted for sCT solutions (50 microm in Dulbecco's modified eagle's medium [DMEM], pH 7.4) from the apical-to-basolateral (A-to-B) direction and B-to-A direction at 37 degrees C and from the A-to-B direction at 4 degrees C. Cell monolayer integrity was monitored by mannitol permeability and transepithelial electrical resistance (TEER) measurements. The permeability coefficients (x 10(-9), cm/sec) for sCT through rat stomach, duodenum, jejunum, ileum, and colon tissues were 0.482 +/- 0.086, 0.718 +/- 0.025, 0.830 +/- 0.053, 1.537 +/- 0.32, and 0.934 +/- 0.15, respectively. The region of maximum sCT permeability is ileum followed by colon, jejunum, duodenum, and stomach. The permeability coefficients (x 10(-6), cm/sec) for sCT through Caco-2 cell monolayer were 8.57 +/- 2.34 (A-to-B, 37 degrees C), 8.01 +/- 1.22 (A-to-B, 4 degrees C), and 6.15 +/- 1.97 (B-to-A, 37 degrees C). The mechanism of its permeation is passive diffusion through the mucosa as determined from the Caco-2 monolayer permeability of sCT. PMID- 15760097 TI - The potential use of Raman mapping to investigate in vitro deposition of combination pressurized metered-dose inhalers. AB - Scanning near-infrared Raman microscopy has been used to map aerosol particulate deposits produced from pressurized metered-dose inhalers (pMDI). A commercially available combination asthma therapy pMDI (Ventide, Allen and Hanbury, UK), containing salbutamol and beclometasone dipropionate, was analyzed by conventional in vitro quantitative analysis and scanning Raman microscopy. Raman maps, taken from Andersen cascade impactor plate stages 3 and 5 (over 100 x 100 microm areas) suggested good correlation with chemical analysis of the respective stages. Scanning Raman microscopy allows visual differentiation between formulation components (not possible using conventional imaging techniques), while potentially allowing chemical quantification. PMID- 15760098 TI - A novel method for the determination of biliary clearance in humans. AB - Biliary excretion is an important route of elimination and the biliary tract is a potential site of toxicity for many drugs and xenobiotics. Quantification of biliary excretion in healthy human volunteers is logistically challenging and is rarely defined during drug development. The current study uses a novel oroenteric tube coupled with a specialized clinical protocol to examine the pharmacokinetics of 99mTechnetium (Tc-99m) mebrofenin, a compound that undergoes rapid hepatic uptake and extensive biliary excretion. A custom-made multilumen oroenteric tube was positioned in the duodenum of healthy human volunteers. Subjects were positioned under a gamma camera and 2.5 mCi of Tc-99m mebrofenin was administered intravenously. Duodenal aspirates, blood samples, and urine were collected periodically for 3 hours. Two hours after Tc-99m mebrofenin administration, the gallbladder was contracted with an intravenous infusion of cholecystokinin-8. Gamma scintigraphy was used to determine the gallbladder ejection fraction in each subject. Total systemic clearance of Tc-99m mebrofenin approximated liver blood flow (Cl(total) 17.3 degrees 1.7 mL/min/kg), and 35% to 84% of the Tc-99m mebrofenin dose was recovered in bile. However, when the data were corrected for the gallbladder ejection fraction, 71% to 92% of the excreted Tc-99m mebrofenin dose was recovered. This novel oroenteric tube and clinical protocol provide a useful method to quantify biliary excretion of xenobiotics in healthy human volunteers. PMID- 15760099 TI - Selective and validated spectrophotometric methods for the determination of nicorandil in pharmaceutical formulations. AB - Two simple and sensitive validated spectrophotometric methods have been described for the assay of nicorandil in drug formulations. Method A is based on the reaction of the drug with phloroglucinol-sulfanilic acid reagent in sulfuric acid medium to give yellow-colored product, which absorbs maximally at 425 nm. Method B uses the oxidative coupling of 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) with DL- 3,4 - dihydroxyphenylalanine (DL-dopa) in the presence of nicorandil as oxidant in sulfuric acid medium to form an intensely colored product having maximum absorbance at 530 nm. Beer's law is obeyed in the concentration range 2.5 to 50.0 and 1.0 to 15.0 microg mL(-1) with methods A and B, respectively. Both methods have been successfully applied for the analysis of drug in pharmaceutical formulations. The reliability and the performance of the proposed methods are established by point and interval hypothesis and through recovery studies. The experimental true bias of all samples is smaller than +/ 2%. PMID- 15760100 TI - Fourier transform infrared spectroscopy for the analysis of neutralizer-Carbomer and surfactant-Carbomer interactions in aqueous, hydroalcoholic, and anhydrous gel formulations. AB - The objective of the present study is to evaluate the polymer-surfactant and polymer-neutralizer interactions in topical aqueous, anhydrous, and hydroalcoholic gel formulations using Fourier transform infrared (FTIR) spectroscopy. The gels were prepared by dispersing Carbomer (Carbopol 980) in water and ethanol for aqueous and anhydrous systems, respectively. Glycerol and propylene glycol were also added to ensure that the compositions of gels closely resembled those used in typical topical gel formulations. Comparisons of the spectra of Carbopol dispersions in aqueous, anhydrous, and hydroalcoholic systems, performed for the first time, show Carbopol-neutralizer and Carbopol surfactant interactions vary depending on the nature of the solvents used for gel formation. Analysis of the spectra of aqueous gel formulations indicates significant presence of ionized carboxyl groups only at higher pH (approximately 8.0). Drying of the aqueous gels causes a shift in the carbonyl stretch band toward higher energy, suggesting changes in polymer-neutralizer interaction. Anhydrous gels exhibit 2 different carbonyl stretch bands: the one at approximately 1653 cm(-1) is related to the carboxyl group that is hydrogen bonded and is akin to hydrous gels; the second one at approximately 1717 cm(-1) is indicative of free carbonyl groups. The carbonyl bands of dried gels appear at different energy levels than the solvated gels. This shift resulting from solvent evaporation, reported for the first time, indicates changes in hydrogen bond characteristics. The results show that FTIR can be a good technique compared with other more time-consuming means of analysis for topical formulations. PMID- 15760102 TI - Formulation design of double-layer in the outer shell of dry-coated tablet to modulate lag time and time-controlled dissolution function: studies on micronized ethylcellulose for dosage form design (VII). AB - The dry-coated tablet with optimal lag time was designed to simulate the dosing time of drug administration according to the physiological needs. Different compositions of ethylcellulose (EC) powder with a coarse particle (167.5 microm) and several fine particles (< 6 microm), respectively, were mixed to formulate the whole layer of the outer shell of dry-coated tablets. The formulations containing different weight ratios of coarse/fine particles of EC powders or 167.5 microm EC powder/excipient in the upper layer of the outer shell to influence the release behavior of sodium diclofenac from dry-coated tablet were also explored. The results indicate that sodium diclofenac released from all the dry-coated tablets exhibited an initial lag period, followed by a stage of rapid drug release. When the mixture of the coarse/fine particles of EC powders was incorporated into the whole layer, the lag time was almost the same. The outer shell broke into 2 halves to make a rapid drug release after the lag time, which belonged to the time-controlled disruption of release mechanism. When the lower layer in the outer shell was composed of 167.5 microm EC powder and the upper layer was formulated by mixing different weight ratios of 167.5 microm and 6 microm of EC powders, the drug release also exhibited a time-controlled disruption behavior. Its lag time might be freely modulated, depending on the amount of 6 microm EC powder added. Once different excipients were respectively incorporated into the upper layer of the outer shell, different release mechanisms were observed as follows: time-controlled explosion for Explotab, disruption for Avicel and spray-dried lactose, erosion for dibasic calcium phosphate anhydrate, and sigmoidal profile for hydroxypropyl methylcellulose. PMID- 15760101 TI - Stabilized dynorphin derivatives for modulating antinociceptive activity in morphine tolerant rats: effect of different routes of administration. AB - Dynorphins, such as dynorphin A(1-13) (Dyn A(1-13)), have been shown to enhance analgesia in morphine-tolerant animals, despite their very short half-life after intravenous administration. The potential use of dynorphins in humans is therefore of interest. This laboratory has recently evaluated the metabolic fate of stabilized dynorphin derivatives. This study was conducted to evaluate whether such stabilized derivatives, ie, [N-Met-Tyr1]-Dynorphin A(1-13) (N-MT Dyn A, stabilized at the N-terminal end) and [N-Met-Tyr1]-Dynorphin A(1-13) amide (N-MT Dyn A amide, stabilized at the C- and N-terminal ends), would enhance the antinociceptive activity of morphine not only after intravenous administration but also after subcutaneous and pulmonary delivery. Intravenous administration of N-MT Dyn A (5 micromol/kg) and N-MT Dyn A amide (5 micromol/kg) to morphine tolerant rats resulted in significantly higher tail-flick latencies than those observed for the saline group. These effects could be observed for up to 2.0 +/- 0.1 hours after intravenous administration of N-MT Dyn A and for up to 3.4 +/- 1.4 hours for N-MT Dyn A amide. The time-averaged effects of both peptides were similar. After pulmonary delivery of the same dose, derivatives remained active. The duration of the effects after pulmonary administration of the amide was 4.4 +/- 2.5 hours while that of N-MT Dyn A was slightly shorter (2.8 +/- 0.9 hours). No effect was observed after subcutaneous administration of N-MT Dyn A. These results suggest that pulmonary delivery of stabilized dynorphin derivatives represents a possible alternative to intravenous administration. PMID- 15760103 TI - Cetirizine from topical phosphatidylcholine-hydrogenated liposomes: evaluation of peripheral antihistaminic activity and systemic absorption in a rabbit model. AB - Cetirizine, an effective, minimally sedating, second-generation H1-antihistamine is widely used orally to treat allergic skin disorders. This study was performed to assess the peripheral H1-antihistaminic activity and extent of systemic absorption of cetirizine from liposomes applied to the skin. Cetirizine was incorporated into small unilamellar vesicles (SUV) and multilamellar vesicles (MLV) prepared using L-alpha-phosphatidylcholine hydrogenated (HPC), and into Glaxal Base (GB) as the control. In a randomized, crossover study, each formulation, containing 10 mg of cetirizine, was applied to the depilated backs of 6 rabbits (3.08 +/- 0.05 kg). Histamine-induced wheal tests and blood sampling were performed before cetirizine application and at designated times for up to 24 hours afterwards. Compared with baseline, histamine-induced wheal formation was suppressed by cetirizine in SUV only at 24 hours, in MLV from 0.5 to 24 hours, and in GB from 0.5 to 8 hours (P < or = .05). Wheal suppression by cetirizine in SUV at 24 hours (91.7% +/- 5.2%) and in MLV from 1 to 24 hours (93.8% +/- 2.2% to 76.2% +/- 6.5%) was greater than in GB (36.5% +/- 7.4% to 60.6% +/- 14.2%) from 1 to 24 hours (P < or = .05). Faster onset, as well as greater and more persistent suppression was obtained from cetirizine in MLV. Plasma cetirizine concentrations from MLV (area under the curve [AUC] of 221.2 +/- 42.3 ng x hr/mL) were lower than from GB (AUC of 248.3 +/- 34.6 ng.hr/mL). In this model, cetirizine from MLV had excellent topical H(1)-antihistamine activity, while systemic exposure was reduced, compared with cetirizine from GB. PMID- 15760104 TI - The back-step method--method for obtaining unbiased population parameter estimates for ordered categorical data. AB - A significant bias in parameters, estimated with the proportional odds model using the software NONMEM, has been reported. Typically, this bias occurs with ordered categorical data, when most of the observations are found at one extreme of the possible outcomes. The aim of this study was to assess, through simulations, the performance of the Back-Step Method (BSM), a novel approach for obtaining unbiased estimates when the standard approach provides biased estimates. BSM is an iterative method involving sequential simulation-estimation steps. BSM was compared with the standard approach in the analysis of a 4 category ordered variable using the Laplacian method in NONMEM. The bias in parameter estimates and the accuracy of model predictions were determined for the 2 methods on 3 conditions: (1) a nonskewed distribution of the response with low interindividual variability (IIV), (2) a skewed distribution with low IIV, and (3) a skewed distribution with high IIV. An increase in bias with increasing skewness and IIV was shown in parameters estimated using the standard approach in NONMEM. BSM performed without appreciable bias in the estimates under the 3 conditions, and the model predictions were in good agreement with the original data. Each BSM estimation represents a random sample of the population; hence, repeating the BSM estimation reduces the imprecision of the parameter estimates. The BSM is an accurate estimation method when the standard modeling approach in NONMEM gives biased estimates. PMID- 15760105 TI - Particle size analysis: AAPS workshop report, cosponsored by the Food and Drug Administration and the United States Pharmacopeia. AB - The concepts of particle engineering and dosage form design have become dominant themes in pharmaceutical manufacturing. This trend is not simply a reflection of the development of new, more sophisticated manufacturing methods of particles or dispersed systems but also recognition of the importance of quality control even in more traditional manufacturing processes. However, the diversity of particle treatments, methods of particle size analysis, expression and interpretation of data, and process applications results in complicated and sometimes confusing criteria for selection, adoption, or relevance of the available techniques. PMID- 15760106 TI - Transporter and ion channel gene expression after Caco-2 cell differentiation using 2 different microarray technologies. AB - mRNA expression profiles had previously been measured in Caco-2 cells (human colonic carcinoma cells) using either custom-designed spotted oligonucleotide arrays or Affymetrix GeneChip oligonucleotide arrays. The Caco-2 cells used were from different clones and were examined under slightly different culture conditions commonly encountered when Caco-2 cells are used as a model tissue for studying intestinal transport and metabolism in different laboratories. In this study, we compared gene expression profiles of Caco-2 cells generated with different arrays to assess the validity of conclusions derived from the 2 independent studies, with a focus on changes in transporter and ion channel mRNA expression levels on Caco-2 cell differentiation. Significant changes in expression levels upon differentiation were observed with 78 genes, with probes common to both arrays. Of these, 18 genes were upregulated and 36 genes were downregulated. The 2 arrays yielded discrepant results for 24 genes, showing significant changes upon differentiation. The results from the 2 arrays correlated well for genes expressed above average levels (r = 0.75, P < 0.01, n = 25) and poorly for genes expressed at low levels (r = 0.08, P > 0.05, n = 25). Overall correlation across the 2 platforms was r = 0.45 (P < 0.01) for the 78 genes, with similar results from both arrays. Despite differences in experimental conditions and array technology, similar results were obtained for most genes. PMID- 15760107 TI - Cryoprotection mechanisms of polyethylene glycols on lactate dehydrogenase during freeze-thawing. AB - The purpose of this study was to explore the cryoprotection mechanisms of high molecular weight polyethylene glycols (PEGs) (eg, PEG 4000 and PEG 8000) on lactate dehydrogenase (LDH). Ultraviolet activity assays, circular dichroism (CD) spectroscopy, gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), (14)C-PEG 4000 labeling and binding, and cryostage microscopic study were conducted. Different molecular weights and concentrations of PEGs in LDH formulations were treated by freeze-thawing. Higher molecular weights and concentrations of PEGs in LDH-PEG formulations obtained better activity and secondary structure recoveries of LDH after freeze-thawing. Insoluble aggregation of LDH was not observed in gel filtration studies. SDS-PAGE results suggested surface characteristic modifications of LDH by the larger molecular weight PEGs. The 14C-PEG 4000 labeling and binding study showed extensive nonspecific interactions between the PEG 4000 and LDH molecules in a concentration-dependent manner. The bound LDH-PEG 4000/free PEG 4000 ratio increased when LDH or PEG 4000 concentrations increased. Cryostage microscopic study showed that PEG 8000 delayed the ice crystallization and eutectic transition of LDH formulation. It appeared that multiple mechanisms were at work during PEGs' cryoprotection of LDH. It was unclear whether the delayed eutectic characteristics of PEGs contributed to LDH cryoprotection. The favorable interaction, rather than preferential exclusion, between LDH and PEGs (eg, 4000) cryoprotected LDH. PMID- 15760108 TI - Etoposide-incorporated tripalmitin nanoparticles with different surface charge: formulation, characterization, radiolabeling, and biodistribution studies. AB - Etoposide-incorporated tripalmitin nanoparticles with negative (ETN) and positive charge (ETP) were prepared by melt emulsification and high-pressure homogenization techniques. Spray drying of nanoparticles led to free flowing powder with excellent redispersibility. The nanoparticles were characterized by size analysis, zeta potential measurements, and scanning electron microscopy. The mean diameter of ETN and ETP nanoparticles was 391 nm and 362 nm, respectively, and the entrapment efficiency was more than 96%. Radiolabeling of etoposide and nanoparticles was performed with Technetium-99m (99mTc) with high labeling efficiency and in vitro stability. The determination of binding affinity of 99mTc labeled complexes by diethylene triamine penta acetic acid (DTPA) and cysteine challenge test confirmed low transchelation of 99mTc-labeled complexes and high in vitro stability. Pharmacokinetic data of radiolabeled etoposide, ETN, and ETP nanoparticles in rats reveal that positively charged nanoparticles had high blood concentrations and prolonged blood residence time. Biodistribution studies of 99mTc-labeled complexes were performed after intravenous administration in mice. Both ETN and ETP nanoparticles showed significantly lower uptake by organs of the reticuloendothelial system such as liver and spleen (P < .001) compared with etoposide. The ETP nanoparticles showed a relatively high distribution to bone and brain (14-fold higher than etoposide and ETN at 4 hours postinjection) than ETN nanoparticles. The ETP nanoparticles with long circulating property could be a beneficial delivery system for targeting to tumors by Enhanced Permeability and Retention effect and to brain. PMID- 15760109 TI - Morphology and buoyancy of oil-entrapped calcium pectinate gel beads. AB - A new emulsion-gelation method to prepare oil-entrapped calcium pectinate gel (CaPG) beads capable of floating in the gastric condition was designed and tested. The gel beads containing edible oil were prepared by either being gently mixed or homogenized an oil phase and a water phase containing pectin, and then extruded into calcium chloride solution with gentle agitation at room temperature. The gel beads formed were then separated, washed with distilled water, and dried at 37 degrees C for 12 hours. A model of the emulsion-gelation process to illustrate the formation of oil-entrapped CaPG beads was proposed. The effect of selected factors, such as type of oil, percentage of oil, and type of pectin on morphology and floating properties was investigated. The oil-entrapped calcium pectinate gel beads floated if a sufficient amount of oil was used. Scanning electron photomicrographs demonstrated very small pores, ranging between 5 and 40 microm, dispersed all over the beads. The type and percentage of oil play an important role in controlling the floating of oil-entrapped CaPG beads. The results suggested that oil-entrapped CaPG beads were promising as a carrier for intragastric floating drug delivery. PMID- 15760110 TI - Neural retina limits the nonviral gene transfer to retinal pigment epithelium in an in vitro bovine eye model. AB - We investigated the permeation of liposomal and polymeric gene delivery systems through neural retina into retinal pigment epithelium (RPE) and determined the roles of various factors in permeation and subsequent uptake of the delivery systems by RPE. Anterior parts and vitreous of fresh bovine eyes were removed. Retina was left intact or peeled away. Complexes of ethidium monoazide (EMA) labeled plasmid DNA and cationic carriers (polyethyleneimine, poly-L-lysine, DOTAP liposomes) were pipetted on the retina or RPE. Two hours later the neural retina was removed, if present, and the RPE cells were detached. Contaminants were removed by sucrose centrifugation, and the RPE cells were analyzed for DNA uptake by flow cytometry. Cellular uptake of FITC-dextrans (molecular weight [mw] 20,000, 500,000 and 2,000,000), FITC-poly-L-lysine (mw 20,000), FITC-labeled oligonucleotide (15-mer), and naked EMA-labeled plasmid DNA was determined after pipetting the solutions on the RPE or neural retina. Location of the fluorescent materials in the retina was visualized with fluorescence microscopy. Neural retina decreased the cellular uptake of DNA complexes by an order of magnitude, the uptake of FITC-dextrans slightly, whereas delivery of polycationic FITC-poly L-lysine to RPE was almost completely inhibited. Neural retina decreased the cellular uptake of FITC-oligonucleotides, while the uptake of uncomplexed plasmid was always negligible. Conclusions from FACS and fluorescence microscopy were similar: delivery of polymeric and liposomal DNA complexes into RPE are limited by the neural retina. This is due to the size and positive charge of the complexes. PMID- 15760111 TI - Optimization of chlorphenesin emulgel formulation. AB - This study was conducted to develop an emulgel formulation of chlorphenesin (CHL) using 2 types of gelling agents: hydroxypropylmethyl cellulose (HPMC) and Carbopol 934. The influence of the type of the gelling agent and the concentration of both the oil phase and emulsifying agent on the drug release from the prepared emulgels was investigated using a 2(3) factorial design. The prepared emulgels were evaluated for their physical appearance, rheological behavior, drug release, antifungal activity, and stability. Commercially available CHL topical powder was used for comparison. All the prepared emulgels showed acceptable physical properties concerning color, homogeneity, consistency, spreadability, and pH value. They also exhibited higher drug release and antifungal activity than the CHL powder. It was found that the emulsifying agent concentration had the most pronounced effect on the drug release from the emulgels followed by the oil phase concentration and finally the type of the gelling agent. The drug release from all the emulgels was found to follow diffusion-controlled mechanism. Rheological studies revealed that the CHL emulgels exhibited a shear-thinning behavior with thixotropy. Stability studies showed that the physical appearance, rheological properties, drug release, and antifungal activity in all the prepared emulgels remained unchanged upon storage for 3 months. As a general conclusion, it was suggested that the CHL emulgel formulation prepared with HPMC with the oil phase concentration in its low level and emulsifying agent concentration in its high level was the formula of choice since it showed the highest drug release and antifungal activity. PMID- 15760112 TI - Development and characterization of biodegradable chitosan films for local delivery of Paclitaxel. AB - Intratumoral and local drug delivery strategies have gained momentum recently as a promising modality in cancer therapy. In order to deliver paclitaxel at the tumor site in therapeutically relevant concentrations, chitosan films were fabricated. Paclitaxel could be loaded at 31% wt/wt in films, which were translucent and flexible. Physicochemical characterization of paclitaxel via thermal, spectroscopic, x-ray diffraction, and electron microscopy techniques revealed information on solid-state properties of paclitaxel as well as chitosan in films. While chitosan was in amorphous form, paclitaxel seemed to be present in both amorphous and crystalline forms in film. The polymeric dispersion of paclitaxel in poloxamer formed fibrous structures generating discontinuities in the film matrix, thereby leading to the introduction of perturbations in the packing arrangement of polymer chains. These films released only 10% to 15% of loaded paclitaxel by a burst effect under in vitro testing conditions, with lysozyme having no effect on the release. However, films softened after implantation in mice and lost integrity over time. The implantable delivery system is not only biodegradable but also well tolerated in vivo and hence, biocompatible as revealed by histological studies. The lack of formulation induced local inflammatory responses of paclitaxel chitosan films suggests a new paradigm for localized chemotherapy based on implantable systems. PMID- 15760113 TI - Single-molecule nanosized polycyclic aromatics with alternant five- and six membered rings: synthesis and optical properties. AB - [structure: see text] A synthetic protocol for a novel family of symmetric polycyclic aromatics with a benzene or cyclooctatetraene (COT) ring as the core and alternant fused benzene and cyclopentadiene rings as branches has been developed. The TiCl(4)-promoted cyclizations construct both planar trimers and tubelike tetramers via the "in situ" generation of the benzene or COT skeleton. The structures of 6b, 7a, 7b, and 10 have been characterized by (1)H and (13)C NMR spectra and MALDI-TOF mass spectroscopy. These polycyclic aromatics also exhibit interesting optical properties. PMID- 15760114 TI - Carbohydrate recognition by porphyrin-based molecularly imprinted polymers. AB - [structure: see text] Porphyrin-based molecularly imprinted polymers (MIPs) were prepared for carbohydrate recognition. A urea-appended porphyrin functional monomer was utilized to provide complementary functionality and quality binding sites throughout the polymer. Each porphyrin-based polymer demonstrates high affinity and differential selectivity for closely related carbohydrates that correlate to the structure of the template used in the imprinting process. PMID- 15760115 TI - Gelation of a highly fluorescent urea-functionalized perylene bisimide dye. AB - [structure: see text] A urea-containing tetraphenoxy-substituted perylene bisimide has been synthesized, and its gelation ability has been studied. This functional dye forms fluorescent organogels in toluene and tetrachloromethane through self-assembly by hydrogen-bonding and pi-pi-stacking interactions. AFM and confocal laser scanning microscopy reveal the formation of fiberlike aggregates. PMID- 15760116 TI - Synthesis of 2,3-substituted tetracenes and evaluation of their self-assembling properties in organic solvents. AB - [structure: see text] Rod-shaped 2,3-di-alkoxytetracenes, soluble in common organic solvents, have been synthesized and studied for their gelling ability in organic solvents and their unusual UV-visible spectroscopic properties. PMID- 15760117 TI - 5,10,15-tris(o-aminophenyl) corrole (TAPC) as a versatile synthon for the preparation of corrole-based hemoprotein analogs. AB - [structure: see text] The atropisomers of 5,10,15-tris(o-aminophenyl) corrole (alphabetaalpha, alphaalphabeta, and alphaalphaalpha) are metastable at room temperature as a result of the low rotational barrier of the o-aminophenyl pickets adjacent to the bipyrrole moiety. Atropisomer enrichment of TAPC was required for the preparation of picket fence, triazacyclononane-capped, and trisimidazole-alphaalphaalpha-corroles. A racemic alpha(2)beta model of cis-A(2)B geometry was also obtained by linking two cis anilines with a short strap and inserting an imidazole tail on the opposite face of TAPC. PMID- 15760118 TI - Enantioselective enol lactone synthesis under double catalytic conditions. AB - [reaction: see text] The reaction of dimedone with 1-(2-alkenoyl)-4-bromo-3,5 dimethylpyrazoles in THF, catalyzed by catalytic amounts of both DBFOX/Ph nickel(II) perchlorate trihydrate and 2,2,6,6-tetramethylpiperidine, in the presence of acetic anhydride in THF produces the corresponding enol lactones in high enantioselectivities through enantioselective Michael additions followed by cyclization with removal of the pyrazole auxiliary. Other related nucleophile precursors can be successfully applied in the enantioselective enol lactone synthesis under the double catalytic conditions. PMID- 15760119 TI - Advances in the universal NMR database approach. 2'-substituted taxanes as probes for an improved protocol of diastereomeric differentiation. AB - [structure: see text] The configuration of the alpha-substituted alpha-hydroxy beta-aminoester moiety in a series of 2'-substituted taxanes was analyzed according to the recently proposed Universal NMR Database (UDB) approach. A critical analysis of the results showed that modifications regarding chemical shift adjustment (so as to render the shifts virtually connectivity independent) were necessary to get consistent stereoassignments in this set of compounds. On this basis, a modified UDB-based strategy, especially tailored to the configurational assignment of densely substituted diastereomeric fragments, is proposed. PMID- 15760120 TI - Convenient method for epoxidation of alkenes using aqueous hydrogen peroxide. AB - [reaction: see text] The complex [Ru(tpy)(pydic)] (1a) is an active catalyst for epoxidation of alkenes by aqueous 30% hydrogen peroxide in tertiary alcohols. The protocol is simple to operate and gives the corresponding epoxides in good to excellent yields. Chiral enantiopure [Ru(tpy)(pydic)] complexes have been synthesized and successfully applied in this procedure. PMID- 15760121 TI - New hemiketal steroid from the soft coral Cladiella sp. AB - [structure: see text] A new hemiketal steroid, named cladiellin A (1), was first isolated from the soft coral Cladiella sp. Its structure was determined by spectroscopic methods and X-ray analysis. Compound 1 easily converted to 1a when NMR spectra were measured in CDCl(3) solution and quickly changed to 1b when pyridine was used. The structures of dehydrated products 1a and 1b were determined by spectroscopic analysis. Bioassay showed that all these three compounds showed antioxidant activity. PMID- 15760122 TI - Surprisingly mild "enolate-counterion-free"pd(0)-catalyzed intramolecular allylic alkylations. AB - [reaction: see text] Palladium-catalyzed intramolecular allylic alkylations of unsaturated EWG-activated amides can take place under phase-transfer conditions or in the presence of a crown ether. These new reaction conditions are milder and higher yielding than those previously reported. A rationalization for such an unexpected result is put forth and validated by DFT-B3LYP calculations. The results suggest cyclization via a counterion-free (E)-enolate TS. PMID- 15760124 TI - The pyrrole approach toward the synthesis of fully functionalized cup-shaped molecules. AB - [reaction: see text] A novel method for the synthesis of new highly functionalized cyclotrimers is described. The method consists of an original synthesis of beta-dibromosubstituted pyrroles, metalation, cycloaddition, and cyclotrimerization. The sequence is highly compatible with common functional groups and allows the construction of cup-shaped molecules functionalized both at the upper and bottom rim. This feature makes the newly formed structures useful scaffolds for the development of supramolecular receptors. PMID- 15760123 TI - Synthesis of adducts of o-quinone metabolites of carcinogenic polycyclic aromatic hydrocarbons with 2'-deoxyribonucleosides. AB - [structure: see text] The first syntheses of the adducts formed in the reactions of o-quinone metabolites of carcinogenic polycyclic aromatic hydrocarbons (BPQ and BAQ) at 2'-deoxyadenosine and 2'-deoxyguanosine sites in DNA are reported. These syntheses entail Pd-catalyzed coupling of protected amine derivatives of catechols with suitably protected halopurine analogues of 2' deoxyribonucleosides. PMID- 15760125 TI - Trifluoromethyl groups in crystal design of 1,4-diphenyl-1,3-butadienes for topochemical [2 + 2] photodimerization. AB - [structure: see text] UV irradiation of the powdered crystalline sample of each of three (E,E)-1,4-di(trifluoromethyl-substituted)phenyl-1,3-butadienes (1-3) was found to yield a single [2 + 2] cycloaddition product in the solid state. Moreover, upon irradiation, the crystalline samples of two (E,E)-1,4 di(trifluoromethyl- and fluorine-substituted)phenyl-1,3-butadienes (4, 5) undergo a similar conversion to afford a [2 + 2] cycloaddition product, respectively. Our observations suggest that trifluoromethyl groups can be used to direct 1,4 diphenyl-1,3-butadiene molecules to form a parallel, offset-stacked orientation suitable for topochemical [2 + 2] cycloaddition. PMID- 15760126 TI - Proximity effects in the palladium-catalyzed substitution of aryl fluorides. AB - [reaction: see text] The aryl fluoride bond has long been considered inert toward Pd-catalyzed insertion reactions. This paper reports for the first time that aryl fluorides bearing an o-carboxylate group can undergo Pd-catalyzed couplings. On the basis of this computational study and subsequent experimental verifications of its predictions, we herein report that such reactions are facilitated by stabilization of the transition state by proximal oxyanions. PMID- 15760127 TI - Chiral Bis-chlorin: enantiomer resolution and absolute configuration determination. AB - [structure: see text] A racemic mixture of the ethane-bridged bis(zinc octaethylchlorin) was successfully resolved for the first time to yield two enantiomers that exhibit substantial CD signals in the regions of chlorin B and Q transitions. The absolute configuration of the corresponding enantiomers was assigned and the origin of its high optical activity was rationalized through a combined spectral, crystallographic, and theoretical analysis. PMID- 15760128 TI - Cycloaddition of delta2-thiazolines and acyl ketenes under acidic conditions results in bicyclic 1,3-oxazinones and not 6-acylpenams as earlier reported. AB - [reaction: see text] Optically active Delta(2)-thiazolines 4 were previously reported to react with acyl Meldrum's acid derivatives 5 under acidic conditions (HCl (g) in benzene) to stereoselectively give 6-acylpenams 1. Recently we have discovered that the structure elucidation of these compounds was incorrect. Thus, we report new data showing that instead of acyl beta-lactams, the optically active isomers 3R,9R-1,3-oxazinones 3a-g are obtained stereoselectively in 38-93% yields. PMID- 15760129 TI - Enantioselective ring opening of meso-epoxides by aromatic amines catalyzed by lanthanide iodo binaphtholates. AB - [reaction: see text] Lanthanide iodo binaphtholates are efficient enantioselective catalysts for the ring opening of meso-epoxides by various aromatic amines. The study of the influence of temperature on the ring opening of cyclohexene oxide by o-anisidine catalyzed by the samarium complex shows an isoinversion effect with the maximum enantiomeric excess at -40 degrees C. Reactions of aniline, o-anisidine, or p-anisidine with five- or six-membered ring epoxides at this temperature allow the preparation of beta-amino alcohols with enantiomeric excesses up to 93%. PMID- 15760130 TI - Synthesis and thermal behavior of new N-heterotolan liquid crystals. AB - [reaction: see text] The synthesis of liquid crystal series 9a-d was achieved using the Buchwald protocol and Sonogashira reaction. PMID- 15760131 TI - Macrocyclic lactam synthesis via a ring expansion reaction: construction of the cripowellin skeleton. AB - [reaction: see text] The cripowellin ring skeleton, a macrocyclic [2.3.5] bicyclic ketolactam, was smoothly generated via construction of a spiro(benzazepin-cyclohexane-1,3-dione) employing oxidative cyclization as a key step and a subsequent ring expansion reaction. PMID- 15760132 TI - Efficient synthesis of water-soluble calixarenes using click chemistry. AB - [reaction: see text] Several water-soluble calix[4]arenes were synthesized via Huisgen 1,3-dipolar cycloaddition between azides and alkynes. Cationic, anionic, and nonionic calixarenes were prepared from a common azidocalixarene intermediate. Azidocalixarenes performed better than alkynylcalixarenes as precursors. The aggregation behavior of the water-soluble calixarenes was studied by (1)H NMR spectroscopy. PMID- 15760133 TI - Synthesis of [1,2,4]triazolo[4,3-alpha]piperazines via highly reactive chloromethyloxadiazoles. AB - [reaction: see text] A concise, modular approach for the synthesis of [1,2,4]triazolo[4,3-alpha]piperazines via condensation of highly reactive chloromethyloxadiazoles with ethylenediamines is described. NMR studies of this reaction provide evidence that suggests a novel activation mechanism for electron deficient chloromethyloxadiazoles. PMID- 15760134 TI - Highly efficient iridium catalyst for asymmetric transfer hydrogenation of aromatic ketones under base-free conditions. AB - [reaction: see text] Catalytic systems generated in situ from the chiral PNNP ligands with iridium or rhodium hydride complexes exhibited excellent catalytic activity and good enantioselectivity in the asymmetric transfer hydrogenation of aromatic ketones without added base. The best result was obtained in the IrH(CO)(PPh(3))(3)-ligand 2 catalytic system with up to 99% yield and 97% ee. PMID- 15760136 TI - Cu(I)-catalyzed enantioselective [3 + 2] cycloaddition reaction of 1 alkylallenylsilane with alpha-imino ester: asymmetric synthesis of dehydroproline derivatives. AB - [reaction: see text] The catalytic, enantioselective [3 + 2] cycloaddition reaction of 1-alkyl-substituted allenylsilanes with alpha-imino ester has been achieved by means of [Cu(MeCN)(4)]BF(4)/(R)-DM-SEGPHOS catalyst to afford silyl substituted dehydroproline derivatives in high yields and enantioselectivities. PMID- 15760135 TI - Bronsted acid-catalyzed highly stereoselective arene-ynamide cyclizations. A novel keteniminium Pictet-Spengler cyclization in total syntheses of (+/-) desbromoarborescidines A and C. AB - [reaction: see text] A Bronsted acid-catalyzed highly stereoselective arene ynamide cyclization is described. These reactions constitute a keteniminium variant of Pictet-Spengler cyclizations, leading to efficient synthesis of nitrogen heterocycles and related alkaloids. Total syntheses of desbromoarborescidines A and C are illustrated here as first applications of this methodology. PMID- 15760137 TI - Synthesis of meso-azulenylporphyrins. AB - [reaction: see text] meso-Azulenylporphyrins 1-4 were prepared by Suzuki-Miyaura coupling or the Ziegler-Hafner method. 1-Azulenyl and 6-azulenyl groups are indeed acting as electron-donating and electron-accepting substituents toward Zn(II) porphyrin. Fluorescence of Zn(II) porphyrin in the dyads is strongly quenched depending upon substitution position of azulene in order of 2 > 1 > 4 > 3. PMID- 15760139 TI - Synthesis of the eastern portion of ajudazol a based on Stille coupling and double acetylene carbocupration. AB - [structure: see text] A strategy for the synthesis of ajudazol A, an unusual, pharmacologically active metabolite from myxobacteria, based on the Stille cross coupling of a 2-stannyl-oxazole with a vinyl iodide unit is described; the vinyl halide unit containing a (Z,Z)-diene was prepared in one pot by the double acetylene carbocupration of a functionalized alkyl cuprate followed by trapping with 2,3-dibromopropene. PMID- 15760138 TI - Modular synthesis of cyclic peptidomimetics inspired by gamma-turns. AB - [reaction: see text] A series of peptidomimetics based on a gamma-turn motif were synthesized using a modular approach, in which N-protected piperidones were reacted with a selection of 2-hydroxyalkyl azides derived from common l-amino acids. Hydrolysis of the initially formed iminium ethers afforded the targeted series of substituted 1,4-diazepin-5-ones. PMID- 15760140 TI - Partial fluorination overcomes herringbone crystal packing in small polycyclic aromatics. AB - [structure: see text] We report the synthesis and characterization of partially fluorinated condensed tetracyclic aromatic compounds. Typical edge-to face/herringbone packing of nonfluorinated analogues is replaced here by columnar stacks with disk planes orthogonal to the columnar axes. Enhanced pi-overlap results with overlaid electron-poor and -rich regions. PMID- 15760141 TI - Enantioselective synthesis of 10-epi-anamarine via an iterative dihydroxylation sequence. AB - [reaction: see text] The enantioselective syntheses of 10-epi-anamarine and 5,10 epi,epi-anamarine have been achieved in 13 to 14 steps. The route relies upon an enantio- and regioselective Sharpless dihydroxylation of either dienoates or trienoates to establish the C-8 to C-11 stereochemistry. A diastereoselective Leighton allylation established the desired C-5 stereochemistry. The route also relies upon a ring-closing metathesis to establish the alpha,beta-unsaturated lactones. PMID- 15760142 TI - Synthesis of pelorol and analogues: activators of the inositol 5-phosphatase SHIP. AB - [reaction: see text] A screening program designed to find new antiinflammatory agents has identified the sponge meroterpenoid pelorol (1) as an in vitro activator of the inositol-5-phosphatase SHIP. Pelorol (1) and several functional group analogues have been synthesized from sclareolide (4). PMID- 15760143 TI - Palladium nanoparticles entrapped in aluminum hydroxide: dual catalyst for alkene hydrogenation and aerobic alcohol oxidation. AB - [reaction: see text] A new aluminum hydroxide-supported palladium catalyst (1) made by a one-pot synthesis through nanoparticle generation and gelation shows a dual catalytic activity for olefinic hydrogenation and aerobic alcohol oxidation. PMID- 15760144 TI - Ruthenium-catalyzed stereoselective intramolecular carbenoid C-H insertion for beta- and gamma-lactam formations by decomposition of alpha-diazoacetamides. AB - [reaction: see text] An operationally simple catalytic system based on [RuCl(2)(p cymene)(2)] was developed for stereoselective cyclization of alpha diazoacetamides by intramolecular carbenoid C-H insertion, and beta-lactams were produced in excellent yields and >99% cis-stereoselectivity. The Ru-catalyzed reactions can be performed without the need for slow addition of diazo compounds and inert atmosphere. With alpha-diazoanilides as substrate, the carbenoid insertion was directed selectively to aromatic C-H bond leading to gamma-lactam formation (>95% yield). PMID- 15760145 TI - Synthesis of the proposed structure of feigrisolide C. AB - [reaction: see text] Possible structures of feigrisolide C were synthesized via ring-closing olefin metathesis reaction of a diester derivative prepared from 8 epi-nonactic acid, but physical characteristics of the synthetic samples did not match with those of the natural sample of feigrisolide C. PMID- 15760146 TI - Synthesis of the core structure of acutumine. AB - [reaction: see text] The tricyclic core of the bioactive natural product acutumine has been synthesized. Key steps include an oxidative phenolic coupling to form a masked o-benzoquinone, an anionic oxy-Cope rearrangement to construct an all-carbon quaternary center, and a Michael-type cyclization to form an amine bearing quaternary carbon. The target compound exists in solution as an enol, in contrast to related compounds that are ketones. A model explaining these observations is presented. PMID- 15760147 TI - Effects of catalyst activation and ligand steric properties on the enantioselective allylation of amines and phenoxides. AB - [reaction: see text] The yields, enantioselectivities, and regioselectivities of the reactions of amines and phenoxides with allylic carbonates in the presence of a metallacyclic iridium catalyst were compared. These data show that both preactivation of the catalyst and the size of the ligand affect the yield, enantioselectivity, and regioselectivity. With the activated catalyst containing a bis-naphthethylamino group, the allylic amination and etherification of a broad range of allylic carbonates occurred in high yields and with high regioselectivities and enantioselectivities. PMID- 15760148 TI - Synthesis of a potent and selective inhibitor of p90 Rsk. AB - [reaction: see text] The synthesis of the naturally occurring kaempferol glycoside SL0101 has been accomplished, as has its biochemical evaluation. SL0101 exhibits selective and potent p90 Rsk inhibitory activity at nanomolar concentrations without inhibiting the function of upstream kinases such as MEK, Raf, or PKC. The synthesis verified the structural assignment of the natural product and has provided access to material sufficient for detailed biological evaluation. PMID- 15760149 TI - Increased structural complexity leads to higher activity: peptides as efficient and versatile catalysts for asymmetric aldol reactions. AB - [reaction: see text] Peptides containing a secondary amine and a carboxylic acid in a specific orientation to each other are presented as highly efficient catalysts for asymmetric aldol reactions: (1) their activity is considerably higher compared to that of proline, and (2) the enantioselectivity of the peptidic catalysts can be changed from (R)- to (S)-selectivity by simple modifications of the secondary structure. PMID- 15760150 TI - Efficient immobilization of a cadmium chemosensor in a thin film: generation of a cadmium sensor prototype. AB - [reaction: see text] The development of an ion-selective chemosensor for Cd(2+) allows generation of a "real-time" sensor. Immobilization of the chemosensor on quartz was achieved in a simple monolayer and in a thin film using a polymer intermediary. As intended, the thin film contains much more chemosensor than the monolayer and provides measurable responses to aqueous Cd(2+) concentrations below 1 microM. Alkali and alkaline earth ions do not interfere with Cd(2+) sensing; Zn(2+) and Cu(2+) are potential interferents. PMID- 15760151 TI - A fast and stable photochromic switch based on the opening and closing of an oxazine ring. AB - [reaction: see text] We have designed a molecular switch based on the photoinduced opening and thermal closing of an oxazine ring. Ultraviolet excitation of this molecule induces the cleavage of a [C-O] bond to form a p nitrophenolate chromophore in less than 10 ns with a quantum yield of ca. 0.1. The photogenerated isomer reverts thermally to the original oxazine within 50 ns. Our photochromic switch survives more than 3000 excitation cycles without decomposing, even in air-saturated solutions. PMID- 15760152 TI - Stereoselective synthesis of tetrahydropyran-4-ones from dioxinones catalyzed by scandium(III) triflate. AB - [reaction: see text] A scandium triflate catalyzed, diastereoselective cyclization between aldehydes and beta-hydroxy dioxinones has been discovered. This process capitalizes on the untapped nucleophilicity of the embedded enol ether within the dioxinone core. The bicyclic compounds from the resulting cyclization can be isolated, or alternatively, alkoxide nucleophiles can be directly added. This in situ addition fragments the dioxinone rings and delivers the 3-carboxy-substituted tetrahydropyran-4-ones in good yields with high levels of diastereoselectivity. PMID- 15760153 TI - Straightforward ring expansion of pyroglutamates to perhydro-1,3-diazepine-2,4 diones. AB - [reaction: see text] Perhydro-1,3-diazepine-2,4-diones are rare and can only be prepared, up to now, by special methods. A new one-step protocol was developed, comprising N-carbamoylation using an isocyanate followed by intramolecular ring expansion. This new methodology provides a straightforward access to this interesting seven-membered skeleton. PMID- 15760155 TI - Highly efficient coupling of beta-substituted aminoethane sulfonyl azides with thio acids, toward a new chemical ligation reaction. AB - [reaction: see text] A highly efficient coupling of protected beta-substituted aminoethane sulfonyl azides with thio acids is reported. In the case of peptide thio acids, this method encompasses a new chemoselective ligation method. Furthermore, the resulting alpha-amino acyl sulfonamides can be alkylated with suitable electrophiles to obtain densely functionalized sulfonamide scaffolds. PMID- 15760154 TI - Synthesis and binding affinities of novel SRIF-mimicking beta-D-glucosides satisfying the requirement for a pi-cloud at C1. AB - [reaction: see text] The synthesis of four bioactive analogues of the somatostatin (SRIF-14) mimetic, beta-d-glucoside (+)-2, in which the C1 indole side chain is replaced with indole surrogates, has been achieved. These congeners, possessing the naphthyl, benzothiophene, benzyl, and benzofuran substituents, were predicted to satisfy the electrostatic requirements of the tryptophan binding pocket of SRIF. Unlike the previously described C4 picolyl and pyrazinyl congeners, these ligands bind the hSST4 receptor. PMID- 15760156 TI - Evidence of the electronic factor for the highly enantioselective catalytic efficiency of Cinchona-derived phase-transfer catalysts. AB - [structure: see text] The Cinchona alkaloid-derived quaternary ammonium salts containing 2'-N-oxypyridine and 2'-cyanobenzene moieties were prepared and evaluated as phase-transfer catalysts in the enantioselective alkylation of glycine imine ester 1. The N-oxypyridine and cyanobenzene moieties might play an important role to form a rigid conformation by coordinating with H(2)O via hydrogen bonding leading to high enantioselectivity (97 approximately >99% ee), which provides evidence of an electronic factor for the high enantioselective catalytic efficiency in phase-transfer alkylation. PMID- 15760157 TI - First approach to the frondosin C ring system via a tandem cyclization/Claisen rearrangement sequence. AB - [reaction: see text] A one-pot tandem 5-exo cyclization/Claisen rearrangement strategy is utilized as the key step in the straightforward construction of the tetracyclic ring system of frondosin C. This reaction is done under microwave irradiation in the presence of catalytic MeLi. PMID- 15760158 TI - Three-component synthesis of alpha,beta-cyclopropyl-gamma-amino acids. AB - [reaction: see text] The multicomponent coupling of alkenylzirconocenes with N diphenylphosphinoyl imines provides rapid access to functionalized C cyclopropylalkylamides which have been readily transformed into alpha,beta cyclopropyl-gamma-amino acids. These novel scaffolds are thus accessible in ca. 8 steps from commercially available alkynes. PMID- 15760159 TI - Substitution and remote protecting group influence on the oxidation/addition of alpha-substituted 1,2-anhydroglycosides: a novel entry into C-ketosides. AB - [reaction: see text] C-Ketosides are valuable intermediates in chemical synthesis and as glycoside mimics. This manuscript describes the efficient generation of these substrates from alpha-alkyl-substituted glycals and an oxidative, C-C bond forming sequence where the choice of C(3) protecting group was critical. PMID- 15760160 TI - Carbohydrate sulfonyl chlorides for simple, convenient access to glycoconjugates. AB - [reaction: see text] The use of carbohydrate sulfonyl chlorides is introduced as a new, facile glycoconjugation method which could find broad applications. We demonstrate the approach by synthesizing a number of glycosylated cholesterol absorption inhibitors which display high inhibitory efficacies in our recently established in vitro assay. Furthermore, we highlight an advantage of the electron-withdrawing nature of the sulfonyl linkage which allowed the synthesis of otherwise unstable azetidine conjugates. PMID- 15760161 TI - Fluoro, alkylsulfanyl, and alkylsulfonyl leaving groups in suzuki cross-coupling reactions of purine 2'-deoxynucleosides and nucleosides. AB - [reaction: see text] Protected 2'-deoxynucleoside and nucleoside derivatives of 6 fluoropurine, 6-(3-methylbutyl)sulfanylpurine, and 6-(3 methylbutyl)ylsulfonylpurine undergo nickel- or palladium-mediated C-C cross coupling with arylboronic acids to give good yields of 6-arylpurine products. PMID- 15760162 TI - Substituted aziridines by lithiation-electrophile trapping of terminal aziridines. AB - [reaction: see text] Regio- and stereoselective deprotonation of N-Bus (Bus = tert-butylsulfonyl)-protected terminal aziridines with lithium 2,2,6,6 tetramethylpiperidide generates a nonstabilized (H-substituted) aziridinyl anion that undergoes in situ or external electrophile trapping under experimentally straightforward conditions to give trans-disubstituted aziridines in good to excellent yields. PMID- 15760164 TI - Transition metal-catalyzed synthesis and reactivity of N-alkenyl aziridines. AB - [reaction: see text] Straightforward methods for palladium-catalyzed alkenylation of aziridines with alkenyl halides and copper-catalyzed alkenylation of aziridines with alkenyl boronic acids have been developed. This methodology offers attractive alternatives to the known methods requiring activated alkenyl halides and acetylenes. A wide variety of N-alkenyl aziridines containing substituents other than electron-withdrawing substituents such as cyano groups and sulfones have been synthesized in good yields. Furthermore, these N-alkenyl aziridines exhibit quite a different reactivity from conventional enamines, as demonstrated by their reactivity. PMID- 15760163 TI - Investigations into the role of ion pairing in reactions of heteroatom substituted cyclic oxocarbenium ions. AB - [reaction: see text] The O-to-C rearrangement of vinyl acetals is used to demonstrate that tight ion pairing is not involved in the stereoselective nucleophilic addition reactions of alkoxy-substituted cyclic oxocarbenium ions. PMID- 15760165 TI - Syntheses of silatranyl- and germatranyluridines. AB - [reaction: see text] Silatranyluridine 1 and germatranyluridine 2 have been prepared in five steps from oxazolinouridine 3 in 27 and 29% yields, respectively. These compounds are novel transition-state analogues (TSAs) for RNA hydrolysis and offer a number of advantages over traditional vanadium- or rhenium based TSAs. Germatrane 2 is completely stable in D(2)O at room temperature, and the half-life of silatrane 1 in D(2)O was found to be >7 days by (1)H NMR spectroscopy. PMID- 15760166 TI - Zinc trimethylsilylamide as a mild ammonia equivalent and base for the amination of aryl halides and triflates. AB - [reaction: see text] We report that Zn[N(SiMe(3))(2)](2) is a mild ammonia equivalent and base for the palladium-catalyzed amination of aryl halides and triflates. In contrast to LiN(SiMe(3))(2), the combination of Zn[N(SiMe(3))(2)](2) and LiCl coupled with aryl halides and triflates containing base-sensitive functionality in high yields. In addition, aryl bromides coupled with aryl and alkylamines with the combination of Zn[N(SiMe(3))(2)](2) and LiCl as base. These aminations occurred without racemization of the enolizable stereocenter of an optically active ester. PMID- 15760167 TI - Spiroacetal biosynthesis: (+/-)-1,7-dioxaspiro[5.5]undecane in Bactrocera cacuminata and Bactrocera oleae (Olive Fruit Fly). AB - [reaction: see text] A biosynthetic scheme rationalizing the formation of (+/-) 1,7-dioxaspiro[5.5]undecane (5) in the fruit fly species Bactrocera cacuminata and Bactrocera oleae (olive fruit fly) is presented. Incorporation studies with deuterium-labeled keto aldehyde (10), 1,5-nonanediol (11), and 1,5,9-nonanetriol (12), and our previous finding that both oxygen atoms of 5 originate from dioxygen, are strongly evidentiary. The racemic condition of the natural spiroacetal 5 is accounted for, and inter alia, it is demonstrated that dihydropyran (18) is not an important intermediate en route to 5. PMID- 15760168 TI - Identification of a tunable site in bryostatin analogs: C20 Bryologs through late stage diversification. AB - [structure: see text] The C20 region of our bryostatin analogs was identified as a nonpharmacophoric site that could be varied to tune analogs for function and physical properties without significantly affecting their binding affinity for PKC. The use of this site in a late-stage diversification strategy has enabled the facile synthesis of a variety of new C20 analogs, all of which retain nanomolar affinity for PKC, in agreement with our pharmacophore hypothesis. PMID- 15760169 TI - Enantioselective direct intermolecular aldol reactions with enantiotopic group selectivity and dynamic kinetic resolution. AB - [reaction: see text] Proline-catalyzed aldol reactions of tetrahydro-4H thipyranone with racemic 1,4-dioxa-8-thia-spiro[4.5]decane-6-carboxaldehyde and with meso/dl 1,4-dioxa-8-thiaspiro[4.5]decane-6,10-dicarboxaldehyde proceed with dynamic kinetic resolution and give single adducts in good yields with excellent ee's. The high enantiotopic group selectivity results from the high intrinsic diastereoface selectivity of the aldehydes. These reactions significantly extend the scope of the enantioselective direct aldol reaction and constitute simple and efficient syntheses of useful tetrapropionate synthons. PMID- 15760170 TI - Preparation of enamides via palladium-catalyzed amidation of enol tosylates. AB - [reaction: see text] A Pd-catalyzed coupling of enol tosylates and amides has been developed. Ligand screening revealed dipf as the most general ligand for this transformation. A variety of enol tosylates were coupled to an array of enamides in 58-97% yield. PMID- 15760171 TI - Pyridine-catalyzed addition of diaryl-1,2-diones to dimethyl butynedioate leading to the formation of 1,2-diaroyl dimethyl maleates via an unprecedented rearrangement. AB - [reaction: see text] Pyridine catalyzes the reaction of 1,2-diaryl diones with dimethyl butynedioate to afford diaroyl maleates. This unprecedented rearrangement involves a unique benzoyl migration and proceeds with complete stereoselectivity. PMID- 15760172 TI - Formation of compound I by photo-oxidation of compound II. AB - [reaction: see text] Compound I is the heme-iron(IV)-oxo porphyrin radical cation formed in peroxidase and catalase enzymes by reaction with hydrogen peroxide. As an alternative to chemical oxidations of porphyrin-iron(III) species, various compound I species were produced by 355 nm laser flash photolysis photooxidation of the corresponding compound II species, porphyrin-iron(IV)-oxo derivatives. The method is demonstrated by production and kinetic studies of the compound I species from 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin-iron, from horseradish peroxidase, and from wild-type horse skeletal myoglobin. PMID- 15760174 TI - Total synthesis of (-)-7-epicylindrospermopsin, a toxic metabolite of the freshwater cyanobacterium aphanizomenonovalisporum, and assignment of its absolute configuration. AB - [reaction: see text] The Z and E nitrones 38 and 39 from condensation of aldehyde 20 with hydroxylamine 36 underwent intramolecular dipolar cycloaddition to give the substituted 1-aza-7-oxobicyclo[2.2.1] heptanes 40 and 41 in a ratio of 2:1, respectively. Reductive N-O bond cleavage of 40 followed by carbonylation gave cyclic urea 47 in which inversion of the secondary alcohol was effected via an oxidation-reduction sequence. After conversion of the p-bromobenzyl ether 50 to azide 54, activation of the cyclic urea as its O-methylisourea and reduction of the azide led to spontaneous cyclization to afford the tricyclic nucleus 59 of cylindrospermopsin. Global deprotection, including hydrolysis of the 2,4 dimethyoxypyrimidine appendage to a uracil, and then monosulfation of the resultant diol 60 afforded a substance identical with natural (-)-7 epicylindrospermopsin (1). The asymmetric synthesis of (-)-7 epicylindrospermopsin defines its absolute configuration as 7S,8R,10S,12S,13R,14S. PMID- 15760175 TI - Synthesis and evaluation of 1-deoxy-D-xylulose 5-phosphoric acid analogues as alternate substrates for methylerythritol phosphate synthase. AB - [structure: see text] Four deoxyxylulose phosphate (DXP) analogues were synthesized and evaluated as substrates/inhibitors for methylerythritol phosphate (MEP) synthase. In analogues CF(3)-DXP (1), CF(2)-DXP (2), and CF-DXP (3), the three methyl hydrogens at C1 of DXP were sequentially replaced by fluorine. In the fourth analogue, Et-DXP (4), the methyl group in DXP was replaced by an ethyl moiety. Analogues 1, 2, and 4 were not substrates for MEP synthase under normal catalytic conditions and were instead modest inhibitors with IC(50) values of 2.0, 3.4, and 6.2 mM, respectively. In contrast, 3 was a good substrate (k(cat) = 38 s(-)(1), K(m) = 227 muM) with a turnover rate similar to that of the natural substrate. These results are consistent with a retro-aldol/aldol mechanism rather than an alpha-ketol rearrangement for the enzyme-catalyzed conversion of DXP to MEP. PMID- 15760176 TI - L-cysteine, a versatile source of sulfenic acids. Synthesis of enantiopure alliin analogues. AB - [reaction: see text] l-Cysteine is a stimulating starting product for the generation of transient sulfenic acids, such as 4, 6, 9, and 15, which add to suitable acceptors, allowing formation of sulfoxides showing a biologically active residue. These sulfoxides are easily isolated in enantiomerically pure form. For instance, N-(tert-butoxycarbonyl)-l-cysteine methyl ester (1a) furnished in few steps sulfenic acid 9a, which was readily converted into (R,S(S))-(2-tert-butoxycarbonylamino-2-methoxycarbonyl-ethylsulfinyl)ethene (22), the methyl ester of Boc-protected nor-alliin. Moreover, the addition of 9a to 2 methyl-1-buten-3-yne has led to a sulfur epimeric and separable mixture of (R)-2 (2-tert-butoxycarbonylamino-2-methoxycarbonyl-ethylsulfinyl)-3-methyl-buta-1,3 dienes 10a and 11a, still possessing a "masked" sulfenic acid function, producible from their cysteine moieties once the dienes have been converted into the desired derivatives. PMID- 15760177 TI - Comparison of carbon-silicon hyperconjugation at the 2- and 4-positions of the N methylpyridinium cation. AB - [structure: see text] N-Methyl-2-trialkylsilylmethylpyridinium cations 6a-c and 4 trialkylsilylmethylpyridinium cations 5a-c were prepared and investigated using (29)Si and (13)C NMR and single-crystal X-ray crystallography. Systematic differences in the (29)Si chemical shifts and (29)Si-(13)C one-bond coupling constants for these cations suggested that the Si-CH(2) bond interacts more strongly at the 2-position of the electron-deficient pyridinium ring than at the 4-position. This result is supported by the X-ray structures of the pyridinium cations 5b and 6b. PMID- 15760178 TI - Neutral homoaromaticity in some heterocyclic systems. AB - [reaction: see text] Neutral homoaromaticity has been evaluated in heterocyclic systems related to the bicyclo[3.2.1]octane skeleton with replacement of CH(2) at C-2 in bicyclo[3.2.1]octa-3,6-diene with X = BH, AlH, Be, Mg, O, S, PH, NH (12); replacement of CH at C-3 in bicyclo[3.2.1]octa-3,6-dien-2-yl anion with PH, S, NH, O (13); and replacement at C-2 and C-3 with N and O (14). Stabilization energies (SE) are evaluated using density functional theory and homodesmotic equations at the B3LYP/6-311+G(3df,2p)//B3LYP/6-31G(d) level for series 12-14. Stabilization energies are compared with diamagnetic susceptibility exaltations, Lambda, CSGT-B3LYP/6-31G(d)//B3LYP/6-31G(d), and nucleus-independent chemical shifts (NICS), GIAO-B3LYP/6-311+G(2d,p)//B3LYP/6-31G(d). Analysis of frontier orbitals and geometries, B3LYP/6-31G(d)//B3LYP/6-31G(d), and proton affinities of 2-azabicyclo[3.2.1]octa-3,6-diene, pyrrole, and related model systems, B3LYP/6 311+G(2d,2p)//B3LYP/6-31G(d), provide complementary evidence supporting the division of the substrates evaluated into antihomoaromatic (12, X = BH, AlH, and Be), nonhomoaromatic (12, X = O, S, NH, PH), and homoaromatic (13, X = S, PH, NH, O and 14 X = ON), with 12 (X = Mg) appearing as transitional between anti- and nonhomoaromatic. PMID- 15760179 TI - Nuclear amination catalyzed by fungal laccases: reaction products of p hydroquinones and primary aromatic amines. AB - [reaction: see text] Nuclear amination of p-hydroquinones with primary aromatic amines was catalyzed by fungal laccases (EC 1.10.3.2) from Trametes spec. and Myceliophthora thermophila. This is the first report of laccase-catalyzed synthesis of aminoquinones. Incubation of two compounds with laccase in the presence of oxygen resulted in the formation of the corresponding monoaminated or diaminated quinones. No hydroquinonoids were formed. Observed differences in the reaction courses for different p-hydroquinones and aromatic amines with different laccases are discussed. PMID- 15760181 TI - Density functional theory predicts the barriers for radical fragmentation in solution. AB - [reaction: see text] N-Methoxypyridyl radicals formed by one-electron reduction of the corresponding cationic heterocycles undergo N-O bond cleavage. Experimental activation free energies for a series of these bond fragmentations are compared to corresponding barriers determined from electronic structure calculations. The DFT barriers agree well with those from experiment, being smaller than the latter values by an average value of ca. 1 kcal/mol, for rate constants varying over almost 3 orders of magnitude, or within ca. 3 kcal/mol over 8 orders of magnitude of rate constant. For a model compound, the B3PW91/6 31+G hybrid density functional method is also found to be in good agreement with the MCSCF-MRMP2 method. One of the reactions is found by DFT to have no minimum for the reactant radical, consistent with a truly barrierless reaction. PMID- 15760180 TI - Family of site-selective molecular optical switches. AB - [reaction: see text] We describe the design, synthesis, and characterization of a family of thiol-reactive optical switches for labeling proteins and other biomolecules. Site-selective introduction of photochromic probes within biomolecules is being used as part of a new approach for optical control of biomolecular interactions and activities within cells. The thiol-reactive photochromic probes described in this report include a spironaphthoxazine and five spirobenzopyrans. The location of the thiol-reactive group on the spirobenzopyran is different for each probe; this feature can be used to control the geometry of the optical switch within a bioconjugate. The photochromes undergo rapid and reversible, optically driven transitions between a colorless spiro (SP) state and a brightly colored merocyanine (MC) state. The MC absorption of a spironaphthoxazine conjugate is red shifted by more than 100 nm compared to the equivalent spirobenzopyran, which may be exploited for the independent control of the MC to SP transition for up to two different spironaphthoxazine and spirobenzopyran conjugates within the same sample. PMID- 15760182 TI - Highly regioselective synthesis of amino-functionalized dendritic polyglycerols by a one-pot hydroformylation/reductive amination sequence. AB - [reaction: see text] Dendritic architectures with neutral core structures and amines groups in the shell are a synthetic challenge, and there is a need for an efficient access. In this paper, highly selective Rh-catalysts are used for sequential hydroformylation/reductive amination of dendritic perallylated polyglycerols 1 with various amines in a one-pot procedure to give dendritic polyamines 3a-e in high yields (73-99%). In all cases, complete conversion of the allyl ether and aldehyde intermediate has been observed. Furthermore, the use of protected amines provides reactive core-shell-type architectures after deprotection. These soluble but membrane filterable multifunctional dendritic polyamines are of high interest as reagents in synthesis or as supports in homogeneous catalysis as well as nonviral vectors for DNA-transfection. PMID- 15760183 TI - Stereoselective recognition of tripeptides guided by encoded library screening: construction of chiral macrocyclic tetraamide ruthenium receptor for peptide sensing. AB - [structure: see text] Molecule sensor 1 is devised by incorporating the reporting unit of ruthenium(II) complex and two recognition motifs of chiral cyclotetraamides on the sidearms. The target binding tripeptides for sensor 1 were readily identified by using an encoded library screening method. This solid phase screening indicated a preferable binding of molecule 1 with d-alanine over the l-isomer. The optical and NMR studies for the binding events of 1 with tripeptides Ac-Ala-Gly-Ala-NHC(12)H(25) in the solution phase showed a consistent trend for the stereoselective recognition of the dd-isomer over the ld-, dl-, and ll-isomers. PMID- 15760184 TI - Transition metal-mediated intramolecular [2+2+2] cycloisomerizations of cyclic triynes and enediynes. AB - [reaction: see text] Nitrogen-containing 15-membered triacetylenic macrocycles known as 1,6,11-tris(arylsulfonyl)-1,6,11-triazacyclopentadeca-3,8,13-triynes (1) and enediynic macrocycles called 1,6,11-tris(arylsulfonyl)-1,6,11 triazacyclopentadeca-3-ene-8,13-diynes (4 and 5) were satisfactorily prepared. [2+2+2] cycloisomerization processes catalyzed by transition metals were tested in the above-mentioned macrocycles. Readily available and familiar cyclotrimerization precatalysts were examined for efficiency. Among them, the RhCl(CO)(PPh(3))(2) complex was found to catalyze the cycloisomerization reaction giving the desired cycloadducts in high yields. PMID- 15760185 TI - Shape-complementarity in the recognition of tricarboxylic acids by a [3+3] polyazacyclophane receptor. AB - [structure: see text] The interaction of a macrocycle containing three trans (1R,2R)-diaminocyclohexane units connected by p-xylene spacers with the isomers 1,3,5-benzenetricarboxylic (1,3,5-BTC), 1,2,4-benzenetricarboxylic (1,2,4-BTC), and 1,2,3-benzenetricarboxylic (1,2,3-BTC) acids and their relevant anions is studied by means of potentiometric and NMR analysis. The interaction is highest for the isomer 1,3,5-BTC, which perfectly fits within the macrocyclic cavity of the host species. The studies have been extended to the triacid 1,3,5 benzenetriacetic observing in this case the effect of a size mismatch between host and guest species. PMID- 15760186 TI - A series of nonpolar thymidine analogues of increasing size: DNA base pairing and stacking properties. AB - [reaction: see text] We describe the properties in DNA of a set of five nonpolar nucleoside mimics in which shape is similar but size is increased gradually. The compounds vary in the size of their exocyclic substituents, which range from hydrogen to iodine, and are designed to test the steric effects of nucleosides, nucleotides, and DNA in biological systems in a systematic way. We describe the conversion of toluene, 2,4-difluorotoluene, 2,4-dichlorotoluene, 2,4 dibromotoluene, and 2,4-diiodotoluene deoxyribosides into suitably protected phosphoramidite derivatives and their incorporation into synthetic DNAs. Studies of their behavior in the context of hexamer and dodecamer duplexes were carried out, with comparison to natural thymine. Thermal melting data with compounds in 5' dangling positions showed that all five compounds stack more strongly than thymine, and all the dihalo-substituted cases stack more strongly than the unsubstituted toluene case. Stacking correlated with surface area and hydrophobicity, both of which increase across the series. In base-pairing studies, all five compounds showed destabilized pairing opposite natural bases (relative to thymine-adenine pairing), as expected. Notably, pairing among the nonpolar base analogues was considerably more stable, and some of the pairs involving the largest analogues showed stability equal to that of a natural thymine-adenine pair. The results establish the base pairing properties of a potentially useful new series of biochemical probes for DNA-protein interactions and also identify a set of new, stable hydrophobic base pairs for designed genetic pairing systems. PMID- 15760187 TI - Selective partial reduction of various heteroaromatic compounds with bridgehead nitrogen via Birch reduction protocol. AB - [reaction: see text] For the first time various heteroaromatic compounds with bridgehead nitrogen, including indolizines, bispyrrolopyrimidines, pyrroloquinolines, pyrroloisoquinolines, and bispyrrolopyrazines, were selectively partially reduced under Birch reduction conditions. It was found that the double bond in the fused heterocycles which possesses the highest LUMO density can be selectively reduced under these conditions. Indolizine 6, containing an ester group at C-6, was reductively alkylated to give dihydroindolizines 8 and 9 possessing a quaternary carbon center in good yield. It was found that ambident substrate 12, under Birch reduction conditions, underwent smooth partial reduction to give 4,5-dihydroquinoline 14 as a sole product with no evidence of reduction of the side chain olefin. It was also shown that electron-rich pyrroloisoquinoline 15, which cannot be reduced via catalytic hydrogenation conditions, was efficiently transformed into its dihydrocounterpart 16 by using the Birch reduction protocol. Finally, it was shown that various fused diazines were smoothly and stereoselectively reduced under Birch reduction conditions to give trans-4,5-disubstituted dihydropyrimidines 30 and 32 in virtually quantitative yields. PMID- 15760188 TI - Regiochemistry of [70]fullerene: preparation of C70(OOtBu)n (n = 2, 4, 6, 8, 10) through both equatorial and cyclopentadienyl addition modes. AB - [reaction: see text] tert-Butylperoxy radicals add to [70]fullerene to form a mixture of adducts C(70)(OO(t)()Bu)(n)() (n = 2, 4, 6, 8, 10). Four isomers were isolated for the bis-adduct with the two tert-butylperoxo groups attached at 1,2 , 5,6-, 7,23-, and 2,5-positions, respectively. Two isomers were isolated for the tetrakis-adduct with the tert-butylperoxo groups located along the equator in C(s)() symmetry and on the side in C(1) symmetry, respectively. Similarly, two isomers were isolated for the hexakis-adducts with a structure related to the tetrakis-adducts, one of which has the cyclopentadienyl substructure. No isomer was detected for the octakis- and decakis-adducts. The C(s)()-symmetric octakis- and C(2)-symmetric decakis-adducts have all the tert-butylperoxo groups located along the equator. The decakis-adduct is the major product under optimized conditions. The compounds were characterized by their spectroscopic data. Chemical correlation through further addition of tert-butylperoxy radicals to isolated pure derivatives confirmed the structure assignment. Mechanisms of the tert-butylperoxy radical addition to C(70) follow two pathways: equatorial addition along the belt and cyclopentadienyl addition on the side. PMID- 15760189 TI - Cis- and trans-strapped calix[4]pyrroles bearing phthalamide linkers: synthesis and anion-binding properties. AB - [reaction: see text] New cis-strapped calix[4]pyrrole derivatives 12, 13, and 19 and trans-strapped systems 14 and 15 bearing isophthalate-derived diamide spacers linked to the tetrapyrrolic core have been synthesized and characterized by spectroscopic means. The anion-binding behavior of these receptors was investigated by proton NMR spectroscopy and isothermal titration calorimetry (ITC). A 2:1 binding stoichiometry was observed under the conditions of NMR analysis but not at the lower concentration regime used for ITC. As gauged from both sets of analyses, these new strapped systems display affinities for halide anions that are enhanced compared to those of normal, unstrapped calix[4]pyrrole. However, contrary to expectations, no size-dependent selectivity for anions is observed as the length of the bridging strap is varied. Such results are interpreted in terms of anion-binding processes that occur outside the central pocket defined by the strap but that still favor strong associations as the result of the increased number of hydrogen-bonding donors the amide groups provide. PMID- 15760190 TI - Routes to N,N'-unsymmetrically substituted 1,3-diketimines. AB - [reaction: see text] A series of novel N,N'-unsymmetrically substituted 1,3 diketimines (3, 12, and 27) have been synthesized from the reaction of exocyclic enaminoketones 8 with amines or metalloenamines (from 13 or 14) with imidoyl thioether 25 or 26. PMID- 15760191 TI - Helix-forming carbohydrate amino acids. AB - [reaction: see text] The solution-phase conformational properties of tetrameric and octameric chains of C-glycosyl alpha-d-lyxofuranose configured tetrahydrofuran amino acids (where the C-2 and C-5 substituents on the tetrahydrofuran ring are trans to each other) were examined using NMR and IR and CD in organic solvents. Studies by NMR and IR demonstrated that in chloroform solution, the tetramer 7 does not adopt a hydrogen-bonded conformation whereas the octamer 10 populates a well-defined helical secondary structure stabilized by 16-membered (i, i - 3) interresidue hydrogen bonds, similar to a pi-helix. Circular dichroism studies in trifluoroethanol are consistent with this conformation for the octamer 10, and also indicate that the tetramer 7 adopts a rigid conformation not stabilized by hydrogen bonds. PMID- 15760192 TI - Bismuth compounds in organic synthesis. A one-pot synthesis of homoallyl ethers and homoallyl acetates from aldehydes catalyzed by bismuth triflate. AB - [reaction: see text] Three one-pot methods for the conversion of aldehydes to homoallyl ethers catalyzed by Bi(OTf)(3).xH(2)O (1 < x < 4) have been developed. The one-pot synthesis of homoallyl ethers can be achieved either by in situ generation of the acetal followed by its reaction with allyltrialkylsilane or by a three-component synthesis in which the aldehyde, trimethylorthoformate or an alkoxytrimethylsilane and allyltrimethylsilane are mixed together in the presence of bismuth triflate (0.1-1.0 mol %). In addition, a three-component synthesis of homoallyl acetates, which is achieved by reacting the aldehyde, acetic anhydride, and allyltrimethylsilane in the presence of bismuth triflate (3.0-5.0 mol %), has been developed. The use of a relatively nontoxic, easy to handle, and inexpensive catalyst adds to the versatility of these methods. PMID- 15760193 TI - Synthetic approaches and total synthesis of natural zoapatanol. AB - [structure: see text] The total synthesis of (+)-zoapatanol utilizing an intramolecular Horner-Wadsworth-Emmons olefination and an enantioselective Sharpless dihydroxylation as the key steps has been achieved. An advanced oxepene intermediate has been obtained by applying a ring-closing metathesis to an unsaturated enol ether. PMID- 15760195 TI - Automatic assignment of absolute configuration from 1D NMR data. AB - [reaction: see text] Opposite enantiomers exhibit different NMR properties in the presence of an external common chiral element, and a chiral molecule exhibits different NMR properties in the presence of external enantiomeric chiral elements. Automatic prediction of such differences, and comparison with experimental values, leads to the assignment of the absolute configuration. Here two cases are reported, one using a dataset of 80 chiral secondary alcohols esterified with (R)-MTPA and the corresponding (1)H NMR chemical shifts and the other with 94 (13)C NMR chemical shifts of chiral secondary alcohols in two enantiomeric chiral solvents. For the first application, counterpropagation neural networks were trained to predict the sign of the difference between chemical shifts of opposite stereoisomers. The neural networks were trained to process the chirality code of the alcohol as the input, and to give the NMR property as the output. In the second application, similar neural networks were employed, but the property to predict was the difference of chemical shifts in the two enantiomeric solvents. For independent test sets of 20 objects, 100% correct predictions were obtained in both applications concerning the sign of the chemical shifts differences. Additionally, with the second dataset, the difference of chemical shifts in the two enantiomeric solvents was quantitatively predicted, yielding r(2) 0.936 for the test set between the predicted and experimental values. PMID- 15760194 TI - Reaction of alpha-(N-carbamoyl)alkylcuprates with propargyl substrates: synthetic route to alpha-amino allenes and delta3-pyrrolines. AB - [reaction: see text] Carbamate deprotonation followed by treatment with CuCN.2LiCl affords alpha-(N-carbamoyl)alkylcuprates which react with propargyl halides, mesylates, tosylates, phosphates, acetates, and epoxides to give alpha (N-carbamoyl) allenes via an anti-S(N)2' substitution process. Propargyl halides, sulfonates, and phosphates give good yields of carbamoyl allenes, while the acetates afford low yields. Propargyl substrates undergo regiospecific S(N)2' substitution in the absence of severe steric hindrance. The alpha-(N-carbamoyl) allenes can be cyclized to 2-oxazolidinones or deprotected to afford the free amines which can be cyclized to Delta(3)-pyrrolines with either AgNO(3) or Ru(3)(CO)(12). PMID- 15760196 TI - Synthesis of 1,10-N,N'-phenanthroline dioxides using HOF.CH(3)CN complex. AB - [reaction: see text] HOF.CH(3)CN, a very efficient oxygen-transfer agent, made readily from F(2), H(2)O, and CH(3)CN, was reacted with various 1,10 phenanthroline derivatives to form the corresponding N,N'-dioxides in good yields and short reaction times. PMID- 15760197 TI - Diastereoselective synthesis and ESR study of 4-phenylDEPMPO spin traps. AB - [reaction: see text] The cis and trans diastereoisomers of 5-(diethoxyphosphoryl) 5-methyl-4-phenylpyrroline N-oxide (4-PhDEPMPOt 8 and 4-PhDEPMPOc 9) were prepared stereoselectively and used as spin traps for hydroxyl and superoxide radicals. The spin adduct formed by reaction of the cis stereoisomer 9with superoxide radical anion exhibited an 8-line ESR spectrum, showing only a reduced alternating line width phenomenon. This spectrum is simpler than the 12-line spectrum of DEPMPO-OOH, which exhibits a strong alternating line width phenomenon. The half-life times of the 4-PhDEPMPOc-OOH and DEPMPO-OOH adducts were of the same order: 14.5 and 15.5 min, respectively. PMID- 15760198 TI - Two-photon induced uncaging of a reactive intermediate. Multiphoton in situ detection of a potentially valuable label for biological applications. AB - [reaction: see text] Two-photon induced Wolff rearrangement of a terphenyl diazoketone 1 was achieved by using focused laser pulses of 532 nm from a Q switched Nd:YAG laser. The nonfluorescent terphenyl diazoketone 1 was transformed into a fluorescent ester derivative 4, which can be detected in situ using the focused laser pulses at 532 nm. Laser power dependence studies show that the Wolff rearrangement is induced by two-photon absorption of the terphenyl diazoketone 1, but suggests that more than two photons of 532 nm are involved (a multiphoton process) in excitation of the ester derivative 4. PMID- 15760199 TI - Hydroxylamines as oxygen atom nucleophiles in transition-metal-catalyzed allylic substitution. AB - [reaction: see text] The viability of hydroxylamines as nucleophiles in transition-metal-catalyzed allylic substitutions was examined. We have found that the oxygen atom of hydroxylamines having an N-electron-withdrawing substituent (also known as hydroxamic acids) acts as a reactive nucleophile. The palladium catalyzed O-allylic substitution of hydroxylamines with allylic carbonate afforded the linear hydroxylamines. The selective formation of the branched hydroxylamines was observed in iridium-catalyzed reaction. Regio- and enantioselective allylic substitution of the unsymmetrical phosphates with hydroxylamines was studied by using the iridium complex of chiral pybox ligand. The aqueous-medium reaction with hydroxylamines proceeded smoothly in the presence of Ba(OH)(2).H(2)O to give the branched products with good enantioselectivities. PMID- 15760200 TI - Spinosyn g: proof of structure by semisynthesis. AB - [reaction: see text] Spinosyn G was isolated in the late 1980s as a minor component from the broth of our potent, fermentation-derived insecticide spinosad. Its structure was then tentatively identified as 5' '-epispinosyn A (3) on the basis of (1)H and (13)C NMR data, but the 4' '-epi compound 4 could not be conclusively ruled out with the data available. Described herein are unambiguous syntheses of both 3 and 4, whereby 3 was proved identical to the natural product. Compound 4 was prepared from intact spinosyn A by a novel F-TEDA-promoted oxidative deamination to the 4' '-ketone 5, stereoselective reduction to the equatorial alcohol 6, and nitrogen incorporation via the axial azide 7. Compound 3 was obtained by coupling spinosyn A 17-pseudoaglycone (9) with the N-protected dihydropyran 16 derived from methyl l-ossaminide (14). This gave an approximately 2:1 mixture of anomeric products 17 with the desired equatorial glycoside predominating, which was then converted to 3 by N-deprotection and methylation. PMID- 15760201 TI - Synthesis of 1,3,5-trisubstituted hydantoins by regiospecific domino condensation/aza-Michael/O-->N acyl migration of carbodiimides with activated alpha,beta-unsaturated carboxylic acids. AB - [reaction: see text] Carbodiimides and suitably activated alpha,beta-unsaturated carboxylic acids react effectively to afford a vast array of 1,3,5-trisubstituted hydantoins by means of a regiospecific domino condensation/aza-Michael/N-->O acyl migration. The reaction works well in very mild conditions (20 degrees C, dichloromethane) with fumaric acid derivatives bearing an electron-withdrawing group in the beta position. Good results have been obtained also with less activated substrates bearing only one electron-withdrawing group in the beta position, using more polar solvents (acetonitrile, DMF), and in the presence of a base (2,4,6-trimethylpyridine). Reactions with asymmetric carbodiimides are generally highly chemo- and regioselective, giving rise to the formation of a single regioisomeric hydantoin. However, asymmetric carbodiimides bearing one alkyl group and one aryl group can produce variable amounts of N-acylurea byproducts. The latter could be easily recovered and transformed into the corresponding hydantoins. A detailed study of the influence of key reaction parameters such as solvent, base, and structure of the reactants on the reaction outcome and mechanism is presented. This methodology is particularly convenient for the synthesis of trifluoromethyl-substituted hydantoins, which could be interesting as bioactive compounds in medicinal chemistry, as well as precursors of the corresponding alpha-amino acids. PMID- 15760203 TI - Asymmetric synthesis of beta-amino carbonyl compounds with N-sulfinyl beta-amino Weinreb amides. AB - [reaction: see text] Diverse organometallic reagents readily add to enantiopure N sulfinyl beta-amino Weinreb amides providing the corresponding, stable, N sulfinyl beta-amino carbonyl compounds in good to excellent yields. This new methodology represents a general solution to the problem of beta-amino carbonyl syntheses, which are important chiral building blocks and constituents of natural products. N-Sulfinyl beta-amino Weinreb amides are prepared by reaction of the potassium enolate of N-methoxy N-methylacetamide with sulfinimines (N-sulfinyl imines) or lithium N,O-dimethylhydroxylamine with N-sulfinyl beta-amino esters. PMID- 15760202 TI - Synthesis of nucleoside boranophosphoramidate prodrugs conjugated with amino acids. AB - [structure: see text] Nucleoside boranophosphates and nucleoside amino acid phosphoramidates have been shown to be potent antiviral and anticancer agents with the potential to act as nucleoside prodrugs. A combination of these two types of compounds results in a boranophosphoramidate linkage between the nucleoside and amino acid. This new class of potential prodrugs is expected to possess advantages conferred by both types of parent compounds. Two approaches, specifically the H-phosphonate and oxathiaphospholane approaches, are described here to synthesize nucleoside boranophosphoramidate prodrugs conjugated with amino acids. The H-phosphonate approach involves a key intermediate, silylated nucleoside amino acid phosphoramidite 6, prepared from a series of reactions starting from nucleoside H-phosphonate in the presence of condensing reagent DPCP. Due to the lengthy procedure and the difficulties in removing DPCP from the final products, we switched to the oxathiaphospholane approach in which the DBU assisted oxathiaphospholane ring-opening process constituted a key step for the generation of nucleoside amino acid boranophosphoramidates 24. We demonstrate that this key step did not cause any measurable C-racemization of boranophosphorylated amino acids 22. Diastereomers of compounds 24a-f were separated by RP-HPLC. An "adjacent"-type mechanism is proposed to explain the diastereomer ratio in the final products obtained via the oxathiaphospholane approach. A tentative assignment of configuration for the diastereomers was carried out based on the mechanism, molecular modeling, and (1)H NMR. Conclusively, the oxathiaphospholane methodology proved to be more facile and efficient than H-phosphonate chemistry in the preparation of the nucleoside amino acid boranophosphoramidate analogues that are promising as a new type of antiviral prodrugs. PMID- 15760204 TI - Phosphine-free hydrazone-Pd complex as the catalyst precursor for a Suzuki Miyaura reaction under mild aerobic conditions. AB - [reaction: see text] Glyoxal bis(N-methyl-N-phenylhydrazone) (1) and its related compounds such as 2-pyridinecarboxaldehyde N-methyl-N-phenylhydrazone (3) were prepared and examined as ligands for the Suzuki-Miyaura cross-coupling reaction of aryl halides and arylboronic acids. We found phosphine-free catalysts, such as Pd(OAc)(2)/hydrazone ligand 1 or 3, to be efficient catalysts for a variety of substrates to produce the coupling products in good yields. PMID- 15760205 TI - Catalytic reductive alkylation of secondary amine with aldehyde and silane by an iridium compound. AB - [reaction: see text] An efficient methodology for the reductive alkylation of secondary amine with aldehyde and Et(3)SiH using an iridium complex as a catalyst has been developed. For example, treatment of dibutylamine with butyraldehyde and Et(3)SiH (a 1:1:1 molar amount of amine, aldehyde, and silane) in 1,4-dioxane at 75 degrees C under the influence of a catalytic amount of [IrCl(cod)](2) gave tributylamine in quantitative yield. In this reaction, no reduction of aldehyde took place. It was found that IrCl(3), which is a starting material for preparation of iridium complexes such as [IrCl(cod)](2), acts as an efficient catalyst for the present reductive alkylation of amine. In addition, a cheaper, easy-to-handle, and environmentally friendly reducing reagent such as polymethylhydrosiloxane (PMHS) in place of Et(3)SiH was also useful. Thus, a variety of secondary amines could be alkylated by allowing them to react with aldehydes and PMHS in the presence of an iridium catalyst to afford the corresponding tertiary amines in good to excellent yields. From the deuterium label experiments, it was revealed that silane and water, generated during the formation of enamine by the reaction of amine and aldehyde, seem to behave as a hydrogen source. The catalytic cycle was discussed. PMID- 15760206 TI - Metalated nitriles: organolithium, -magnesium, and -copper exchange of alpha halonitriles. AB - [reaction: see text] alpha-Halonitriles react with alkyllithium, organomagnesium, and lithium dimethylcuprate reagents generating reactive, metalated nitriles. The rapid halogen-metal exchange with alkyllithium and Grignard reagents allows selective exchange in the presence of reactive carbonyl electrophiles, including aldehydes, providing a high-yielding alkylation protocol. Lithiated and magnesiated nitriles react with propargyl bromide by S(N)2 displacement whereas organocopper nitriles react by S(N)2' displacement, correlating with the formation of a C-metalated nitrile. PMID- 15760207 TI - Cycloaddition chemistry of 2-vinyl-substituted indoles and related heteroaromatic systems. AB - [reaction: see text] The intramolecular Diels-Alder cycloaddition reaction (IMDAF) of several N-phenylsulfonylindolyl-substituted furanyl carbamates containing a tethered pi-bond on the indole ring were examined as an approach to the iboga alkaloid catharanthine. Only in the case where the tethered pi-bond contained two carbomethoxy groups did the [4 + 2]-cycloaddition occur. Push-pull dipoles generated from the Rh(II)-catalyzed reaction of diazo imides, on the other hand, undergo successful intramolecular 1,3-dipolar cycloaddition across both alkenyl and heteroaromatic pi-bonds to provide novel pentacyclic compounds in good yield and in a stereocontrolled fashion. The facility of the cycloaddition was found to be critically dependent on conformational factors in the transition state. Ligand substitution in the rhodium(II) catalyst markedly altered the product ratio between [3 + 2]-cycloaddition and intramolecular C-H insertion. The variation in reactivity reflects the difference in electrophilicity between the various rhodium carbenoid intermediates. Intramolecular C-H insertion is enhanced with the more electrophilic carbene generated using Rh(II) perfluorobutyrate. PMID- 15760208 TI - Dimethyl carbonate as an ambident electrophile. AB - [reaction: see text] The features of various anions having different soft/hard character (aliphatic and aromatic amines, alcohoxydes, phenoxides, thiolates) are compared with regard to nucleophilic substitutions on dimethyl carbonate (DMC), using different reaction conditions. Results are well in agreement with the Hard Soft Acid-Base (HSAB) theory. Accordingly, the high selectivity of monomethylation of CH(2) acidic compounds and primary aromatic amines with DMC can be explained by two different subsequent reactions, which are due to the double electrophilic character of DMC. The first step consists of a hard-hard reaction and selectively produces a soft anion, which, in the second phase, selectively transforms into the final monomethylated product, via a soft-soft nucleophilic displacement (yield >99% at complete conversion, using DMC as solvent). PMID- 15760209 TI - Total synthesis of (+)-crocacin D. AB - [structure: see text] The total synthesis of (+)-crocacin D is described. The convergent asymmetric synthesis relies on the use of a Stille cross-coupling between an (E)-vinyl stannane with an (E)-vinyl iodide to establish the (E,E) dienamide moiety followed by a mild and efficient copper-catalyzed coupling between (+)-crocacin C and a (Z)-vinyl iodide to establish the challenging (Z) enamide function. PMID- 15760210 TI - A stereoselective and atom-efficient approach to multifunctionalized five- and six-membered rings via a novel michael-initiated intramolecular Diels-Alder furan reaction. AB - [reaction: see text] A variety of key precursors to the intramolecular Diels Alder reaction of furan diene (IMDAF) have been prepared via a very facile 1,4 addition of O-, S-, N-, and C-centered nucleophiles possessing unsaturated tether to beta-furyl nitroethylene. Subsequent IMDAF reaction of the 1,4-adducts, carried out under thermal conditions, provided five- and six-membered carbocycles and heterocycles fused to an easily cleavable oxabicycloheptene moiety. The structure and stereochemistry of the cycloadducts were determined by 2D-NMR experiments and further confirmed by X-ray crystallography. The salient features of the strategy include high degree of stereoselectivity (>80:20) in the cycloaddition, atom and step economy, and generation of multiple chiral centers and functionalities. The feasibility of the cleavage of the oxa bridge in the cycloadducts to afford novel multifunctional molecules has also been demonstrated. PMID- 15760211 TI - Radical-mediated carboxylation of alkyl iodides with [11C]carbon monoxide in solvent mixtures. AB - [reaction: see text] [carboxyl-(11)C]Carboxylic acids were prepared from alkyl iodides via photoinitiated radical reactions using 10(-)(8) mol of [(11)C]carbon monoxide in binary and ternary homogeneous solvent mixtures. Short- (isobutyric), medium-, and long-chain saturated fatty acids (heptadecanoic) were labeled with isolated decay-corrected radiochemical yields ranging from 55% to 70% in 5-7-min reactions. The conversion of [(11)C]carbon monoxide to products reached 80-90%. To obtain good yields in the reactions performed in water-acetonitrile and water THF mixtures, the addition of tetrabutylammonium hydroxide or potassium hydroxide was essential. The carboxylation was efficient for primary and secondary alkyl iodides. The carboxylation of tertiary iodides was feasible for 1-iodoadamantane but not for tert-butyl iodide. The dependence of the radiochemical yields on reaction time, photoirradiation conditions, and organic and inorganic additives was studied. The method provides a one-step route to [carboxyl-(11)C]carboxylic acids; traditional methods, in contrast, would require several steps. For example, using the devised reaction conditions, 3.19 GBq of purified [1 (11)C]1,10-decanedicarboxylic acid (specific radioactivity 188 GBq/mumol) was obtained within 35 min of the end of 10 muAh bombardment. (1-(13)C)4 Phenylbutyric acid was synthesized using ((13)C)carbon monoxide for identifying the labeling position with (1)H and (13)C NMR. PMID- 15760212 TI - The first total synthesis of (-)-methyl barbascoate. AB - [reaction: see text] The neo-trans-clerodane natural product, (-)-methyl barbascoate 1, has been synthesized for the first time starting from the known ketone 6 derived from (R)-(-)-Wieland-Miescher ketone analogue 5. PMID- 15760213 TI - An efficient, PIFA mediated approach to benzo-, naphtho-, and heterocycle-fused pyrrolo[2,1-c][1,4]diazepines. An advantageous access to the antitumor antibiotic DC-81. AB - [reaction: see text] The synthesis of a series of optically pure benzo-, naphtho , and heterocycle-fused pyrrolo[2,1-c][1,4]-diazepin-5,11-dione derivatives starting from l-proline methyl ester is presented. The synthetic plan includes an aroylation step at the proline nitrogen followed by transformation of the ester residue into a N-methoxyamide group. The subsequent key cyclization step embraces the PIFA mediated formation of a N-acylnitrenium intermediate and its succeeding intramolecular trapping by the aromatic ring. The presented general approach solves the need of starting from not very accessible amino (or a related functionality) aromatic starting materials, and its effectiveness is demonstrated in the synthesis of the antitumor antibiotic DC-81. PMID- 15760214 TI - Gold-catalyzed reactions of 2-alkynyl-phenylamines with alpha,beta-enones. AB - [reaction: see text] The gold-catalyzed reaction of 2-alkynyl-phenylamines with alpha,beta-enones represents a new general one-pot entry into C-3-alkyl-indoles by sequential reactions. Gold-catalyzed sequential cyclization/alkylation, N alkylation/cyclization, or N-alkylation/cyclization/alkylation reactions leading to different indoles can be directed by changing the 2-alkynyl-phenylamine 1/alpha,beta-enone 3 ratio and the reaction temperature. Unusual gold-catalyzed rearrangement reaction of indoles are observed at 140 degrees C. New gold catalyzed formation of propargyl-alkyl ether under mild conditions and the hydration reaction of N-acetyl-2-ethynyl-phenylamine are reported. PMID- 15760215 TI - Direct access to terpyridine-containing polyazamacrocycles as photosensitizing ligands for Eu(III) luminescence in aqueous media. AB - [reaction: see text] The synthesis of new 18-membered hexaazamacrocycles containing a functionalized 2,2':6',2' '-terpyridine moiety as part of the cyclic backbone and three acetate pendant arms is described. The reported synthetic procedure is based on the use of an efficient metal template ion effect which controls the macrocyclization step. This procedure is compatible with some functional groups present in the macrocyclic structure. The photophysical properties of the Eu(III) complexes derived from these ligands were examined in aqueous solutions. Their luminescence lifetimes (tau approximately 1 ms) and quantum yields (13% < Phi < 18%) on one hand, their high kinetic inertness on the other hand, and the presence of additional functionality allowing their covalent conjugation to biomolecules seriously nominate these complexes as very promising candidates for luminescent labeling of biological materials. PMID- 15760216 TI - Azulene-substituted aromatic amines. synthesis and amphoteric redox behavior of N,N-Di(6-azulenyl)-p-toluidine and N,N,N',N'-tetra(6-azulenyl)-p-phenylenediamine and their derivatives. AB - [reaction: see text] N,N-Di(6-azulenyl)-p-toluidine (1a) and N,N,N',N'-tetra(6 azulenyl)-p-phenylenediamine (2a) and their derivatives with 1,3 bis(ethoxycarbonyl) substituents on each 6-azulenyl group (1b and 2b) were prepared by Pd-catalyzed amine azulenylation and characterized as a study into new aromatic amines for multistage amphoteric redox materials. The redox behavior of each compound was characterized by cyclic voltammetry. These compounds undergo facile reduction to stable anion radicals and dianion diradicals owing to the resonance stabilization between the 6-azulenyl groups and exhibit electrochemical oxidation depending on the amine subunits. The ESR measurement of anion radicals and a dianion diradical generated by the electrochemical reduction of amine 1b and diamine 2b revealed that the unpaired electron of these radicals delocalizes over the entire azulene ring including the central nitrogen atoms. UV-vis spectral analysis of amines 1a,b and diamines 2a,b, taken during the electrochemical reduction, exhibited a gradual decrease of the absorption bands of the neutral species along with an increase of the new absorption maxima at 625, 605, 640, and 610 nm, respectively, with the development of well-defined isosbestic points at 502, 562, 478, and 545 nm, respectively. As indicated by a combined ESR and UV-vis spectral study, the species giving rise to the new absorption maxima are concluded to be the generation of anion radicals and dianion diradicals of aromatic amines and diamines with high thermodynamic stability. PMID- 15760217 TI - Structure, bonding, and solvation of lithium vinylcarbenoids. AB - [reaction: see text] Molecular modeling was used to determine the structure of lithium vinylcarbenoids in the gas phase and in THF solution. Solvent effects were modeled by microsolvation with explicit THF ligands on each of the lithium atoms. The carbenoid geometries are dependent on the heteroatom and on solvation. The calculations predict 1-chlorovinyllithium and 1-bromovinyllithium to be a mixture of monomer and dimer at 200 K and mostly monomer at higher temperatures, whereas the 1-fluoro-, 1-methoxy-, and 1-dimethylaminovinyllithium are predicted to be dimeric in solution. PMID- 15760218 TI - Highly congested nondistorted diheteroarylnaphthalenes: model compounds for the investigation of intramolecular pi-stacking interactions. AB - [structure: see text] The rigid, highly congested structure of 1,8 diacridylnaphthalenes has been studied in solution and in the solid state. The unique geometry of these compounds forces the acridyl rings to undergo face-to face interactions while rendering T-shaped orientations and face-to-edge interactions impossible. Crystallographic analysis shows that splaying between the heteroaryl rings decreases while twisting between the cofacial rings increases as the acridyl nitrogens of the 1,8-diacridylnaphthalene framework are subsequently oxidized. The peri-acridyl rings are slightly splayed but remain perfectly planar in all cases. The significant decrease in splaying indicates enhanced pi-pi-attraction between the electron-rich acridyl N-oxide moieties, which is in agreement with recently reported symmetry-adapted perturbation theory calculations. The pi-stacking and the molecular geometry between the acridyl rings observed in the solid state have been confirmed through in-solution studies showing characteristic proton NMR upfield shifts and optical properties indicative of static intramolecular arene-arene interactions. Acridyl protons located directly above the adjacent aryl moiety as a consequence of twisting between the heteroaryl rings were identified by COSY NMR measurements and found to intrude into the pi-cloud and diamagnetic ring current of the neighboring acridine. Different shapes and strong red shifts of the fluorescence emission maxima of the diacridylnaphthalenes in comparison to parental acridyl monomers have been attributed to static excimer emission. PMID- 15760219 TI - 1,3-dipolar cycloaddition reactions of porphyrins with azomethine ylides. AB - [reaction: see text] The behavior of porphyrins as dipolarophiles in 1,3-dipolar cycloadditions with azomethine ylides was studied. Depending on the nature of the substituent groups on the porphyrin macrocycles, the reaction can give monoadducts (chlorins) or bisadducts (isobacteriochlorins and bacteriochlorins). When a large excess of azomethine ylide is used, trisadducts can also be obtained. Mixed isobacteriochlorin derivatives were prepared from the reaction of azomethine ylides with the chlorin monoadducts previously obtained via Diels Alder reactions. PMID- 15760220 TI - Design of a chiral mesoporous silica and its application as a host for stereoselective di-pi-methane rearrangements. AB - [reaction: see text] A chiral periodic mesoporous organosilica (ChiMO) was prepared, in which a 1,2-bis-(ureido)cyclohexyl linker (38 wt %) is introduced into the walls of the hybrid organic-inorganic material. This silica was used as a host for 11-formyl-12-methyldibenzobarrelene (2), and the stereoselectivity of the di-pi-methane rearrangement of 2 within this host-guest complex was studied. At low conversions, the only product was the corresponding dibenzosemibullvalene. An enantiomeric excess of 24% at 11% conversion was obtained using the ChiMO as host. These values compare well with those achieved using a system based on conventional faujasites (LiY and NaY) as hosts incorporating a chiral auxiliary. We tested S-phenylglycine, S-proline, S-camphanic acid, and S-mandelic acid as chiral auxiliaries. In contrast to the behavior of the ChiMO material, adsorption of dibenzobarrelene in purely siliceous mesoporous MCM-41 silica (3.2 nm pore size) containing ephedrine failed, a failure that can be explained as arising from the large internal silanol population and high hydrophilicity of the siliceous MCM-41 sample. PMID- 15760221 TI - Heterocyclic rearrangements in constrained media. A zeolite-directed photorearrangement of 1,2,4-oxadiazoles. AB - [reaction: see text] The first intrazeolite-photoinduced rearrangement of a five membered heterocycle is reported. A completely different behavior compared to solution irradiations has been observed. The zeolite's role in directing the photoreaction of 3-phenyl-1,2,4-oxadiazoles toward the formation of the corresponding 1,3,4-oxadiazoles in a ring contraction-ringexpansion route is discussed. PMID- 15760222 TI - General approach to glycosidase inhibitors. Enantioselective synthesis of deoxymannojirimycin and swainsonine. AB - [reaction: see text] Deoxymannojirimycin (2) and swainsonine (4) have been synthesized from each enantiomer of the same bicyclic carbamate precursor 7. The key intermediate was prepared by a simple and efficient three-step synthesis involving RCM of the diene 8, which in turn is easily accessible in any configuration from enantiomerically enriched 2,3-epoxy-4-penten-1-ol 9. PMID- 15760223 TI - Asymmetric allylboration of alpha,beta-enals as a surrogate for the enantioselective synthesis of allylic amines and alpha-amino acids. AB - [reaction: see text] Optically pure allylic amines have been synthesized from alpha,beta-unsaturated aldehydes via allylboration with (-)-B allyldiisopinocampheylborane, followed by Overman rearrangement. By incorporating crotyl and alkoxyallylboration, functionalization at delta-position was readily accomplished. By applying this methodology, the synthesis of several chiral alpha amino acids has been achieved. PMID- 15760224 TI - Microwave-promoted Suzuki reactions of aryl chlorides in aqueous media. AB - [reaction: see text] The microwave-promoted Suzuki coupling reaction of aryl chlorides with boronic acids performed in an aqueous media was studied using the air- and moisture-stable catalyst POPd2 (dihydrogen di-mu-chlorodichlorobis(di tert-butylphosphinito-kappaP)dipalladate (2-)). This catalyst system under microwave conditions (150 degrees C, 15 min) provided coupled products with yields ranging from 64% to 99%. This method tolerated a variety of substituents and sterically hindered substrates. PMID- 15760226 TI - Ionic liquid-immobilized quinuclidine-catalyzed Morita-Baylis-Hillman reactions. AB - [reaction: see text] The ionic liquid-bound quinuclidine catalyzed Baylis-Hillman reactions were investigated. The IL-supported catalyst showed equally good catalytic activity as compared with its nonimmobilized counterpart. The corresponding Baylis-Hillman adducts were obtained in moderate to high yields in all the cases tested. The IL-supported quinuclidine can be readily recovered and reused six times without significant loss of catalytic activity. PMID- 15760225 TI - BF3-mediated additions of organolithiums to ketimines: X-ray crystal structures of BF3-ketimine complexes. AB - [reaction: see text] Additions of lithium acetylides and n-BuLi to N-alkyl ketimines mediated by BF(3)-Et(2)O in THF afford hindered tert-alkylamines in moderate to good yields. Stereochemical results and crystal structures of three BF(3)-imine complexes suggest that allylic strain strongly influences conformation and may be an important determinant of reactivity and selectivity. PMID- 15760227 TI - Radical alpha-C-H hydroxyalkylation of ethers and acetal. AB - [reaction: see text] Epsilonthers and an acetal were found to undergo direct intermolecular addition to aldehydes under the Et(3)B/air conditions. This study presents a very unique and simple means for the radical alpha-C-H hydroxyalkylation of oxygen-containing compounds. PMID- 15760228 TI - Synthesis and palladium-catalyzed coupling reactions of enantiopure p-bromophenyl methyl sulfoximine. AB - [reaction: see text] The asymmetric synthesis and chemical modification of p bromophenyl methyl sulfoximine (2) is described. Starting from p-bromophenyl menthyl sulfinate (5), enantiopure 2 can be obtained in a short reaction sequence involving a well-established substitution reaction followed by stereospecific imination with O-mesitylenesulfonylhydroxylamine (MSH). Palladium-catalyzed Buchwald/Hartwig, Suzuki, and Stille coupling reactions allow a broad variation of the sulfoximine aryl group, which is otherwise difficult to achieve. The incorporation of a p-morpholino-substituted derivative into a pseudotripeptide demonstrates the applicability of the novel sulfoximine derivatives. PMID- 15760229 TI - Efficient preparation of nitrosoarenes for the synthesis of azobenzenes. AB - [reaction: see text] Reaction conditions are described for the oxidation of anilines furnishing nitrosoarenes and the synthesis of unsymmetrically substituted azobenzenes. In a comparative study, the catalytic oxidation of methyl 4-aminobenzoate by hydrogen peroxide was investigated, and SeO(2) proved to be superior or equal to methyl trioxorhenium (MTO) and Na(2)WO(4), respectively. Nevertheless, the application of the inexpensive, environmentally friendly, Oxone in a biphasic system proved to be more efficient, and a variety of useful nitrosoarenes for the synthesis of azo compounds were prepared in high yield and purity on a large scale. PMID- 15760230 TI - Highly efficient and direct heterocyclization of dipyridyl ketone to N,N bidentate ligands. AB - [reaction: see text] Reaction of various aromatic aldehydes with 2,2'-dipyridyl ketone and ammonium acetate in hot acetic acid provides ready access to a series of substituted 1-pyridylimidazo[1,5-a]pyridines, a class of ligands possessing an N,N-bidentate feature, in good yields. PMID- 15760231 TI - Synthesis of heterocyclic allenes via palladium-catalyzed hydride-transfer reaction of propargylic amines. AB - [reaction: see text] Propargylic diisopropylamines containing heterocycles, which were prepared readily from heterocyclic bromides and propargyldiisopropylamine by the Sonogashira coupling reaction, underwent the allene transformation reaction in the presence of Pd(2)(dba)(3).CHCl(3) catalyst (2.5 mol %) and 1,2 bis[bis(pentafluorophenyl)phosphino]ethane (10 mol %) at 100 degrees C in CHCl(3), giving the corresponding heterocyclic allenes in good to high yields via the palladium-catalyzed hydride-transfer reaction. PMID- 15760232 TI - Microwave-assisted synthesis of isonitriles: a general simple methodology. AB - [reaction: see text] A facile conversion of formamides to isonitriles under very mild conditions and microwave irradiation is described. This simple and efficient method has been applied for the synthesis of both aliphatic and aromatic isonitriles in high yields. PMID- 15760233 TI - A practical method for the synthesis of 2-alkynylpropenals. AB - A general method for the preparation of 2-alkynyl acroleins is described beginning with vinyl iodide 5 and involving a combination of Sonogashira coupling and Dess-Martin oxidation. Critical to the success of this approach is the use of a special workup procedure for the oxidation step. The resultant enynals participate in a variety of addition reactions including aldol condensations and reactions with organolithium compounds. PMID- 15760234 TI - [4+2] Cycloaddition of 2-substituted 1,2-dihydropyridines with nitrosobenzene: asymmetric synthesis of trans-2-substituted 3-amino-1,2,3,6-tetrahydropyridines. AB - [reaction: see text] A new methodology for the stereoselective synthesis of trans 2-substituted 3-amino-1,2,3,6-tetrahydropyridines is reported. The preparation of these 3-aminopiperidines is achieved by cycloaddition of nitrosobenzene with 2 substituted 1,2-dihydropyridines followed by chemoselective reduction of the cycloadducts. Enantioenriched 1,2-dihydropyridine derivatives are easily prepared from pyridine and a chiral amide following a previous report from our laboratories. Moreover, the in situ hydrogenation of these cycloadducts over palladium in a solution of hydrogen chloride in methanol led to tetrahydropyrroloimidazoles. PMID- 15760235 TI - An efficient chemoenzymatic approach to (S)-gamma-fluoroleucine ethyl ester. AB - [reaction: see text] An asymmetric synthesis of (S)-gamma-fluoroleucine ethyl ester 1 is described. The key transformation involves a lipase-catalyzed dynamic ring-opening of 2-(3-butenyl)azlactone 7b with EtOH to give amide ester (S)-6b in 84% enantiomeric excess. Removal of the N-pentenoyl group with N,N' dibromodimethylhydantoin in the presence of trifluoroacetic acid afforded the titled compound, which was isolated as its hydrogen sulfate salt in 75% yield and >97% ee. PMID- 15760236 TI - A new synthesis of 2-substituted pyridines via aluminum chloride induced heteroarylation of arenes and heteroarenes. AB - [reaction: see text] We herein report a new synthesis of 2-(hetero)aryl substituted pyridines via heteroarylation of arenes/heteroarenes through AlCl(3) induced C-C bond-forming reactions. 2-Halopyridines bearing an electron withdrawing group were reacted with a number of (hetero)arenes to give 2 aryl/heteroaryl-substituted pyridines in good yields. PMID- 15760237 TI - One-pot sequential synthesis of acetoxylated [60]fullerene derivatives. AB - [reaction: see text] The reaction of [60]fullerene with 4-substituted phenylhydrazine hydrochlorides in refluxing chlorobenzene under aerobic conditions afforded 1-(4-substituted phenyl)-1,2-dihydro[60]fullerenes, which could be subsequently oxidized to 1-acetoxyl-4-aryl-1,4-dihydro[60]fullerenes by manganese(III) acetate dihydrate in one pot. The transformation of ArC(60)-H to ArC(60)-OAc has been realized with Mn(OAc)(3).2H(2)O for the first time. PMID- 15760238 TI - Synthesis of a 4,6-disubstituted dibenzofuran beta-sheet initiator by reductive radical arylation of benzene. AB - [reaction: see text] Tributyltin hydride mediated addition of 3 iodosalicylaldehyde to benzene in the presence of catalytic benzeneselenol affords (1,4-cyclohexadien-3-yl)salicylaldehyde. Homologation of the aldehyde group is followed by cycloetherification with dimethyl dioxirane to give a 4,6 disubstituted tetrahydrodibenzofuran. Adjustment of oxidation states and introduction of a second chain by Wittig olefination affords the beta-sheet initiator, ethyl 4-(2-tert-butoxycarbonylaminoethyl)-6-dibenzofuranpropanoate. PMID- 15760239 TI - Glycal scavenging in the synthesis of disaccharides using mannosyl iodide donors. AB - [reaction: see text] High mannose glycans composed of alpha (1-->2) and alpha (1- >6) branched sugars are important components of the HIV-associated envelope glycoprotein, gp120. These substructures can be efficiently prepared in solution from glycosyl iodide precursors requiring only a slight excess of the iodide donor, which offers advantages over solid-phase methods that require more than 5 equiv of donor. During the reaction, excess iodide is converted to a glycal that is not easily separated from the desired disaccharide. To overcome this difficulty, a phase-trafficking methodology that relies upon nucleophilic interception of the 1,2 anhydrosugar resulting from oxidation of the glycal has been developed. PMID- 15760240 TI - Silver(I)-carbene complexes/ionic liquids: novel N-heterocyclic carbene delivery agents for organocatalytic transformations. AB - [reaction: see text] N-Heterocyclic carbene (NHC) complexes with silver were investigated as sources of unsaturated NHC carbene catalysts via thermal decomposition. The NHC complex (1-ethyl-3-methylimidazol-2-ylidene)silver(I) chloride is an ionic liquid, and was found to catalyze the ring-opening polymerization of lactide at elevated temperatures to give narrowly dispersed polylactide of predictable molecular weight. Silver-carbene complexes can also be used for the catalysis of small molecule transesterification reactions. Thermolysis of the silver complexes in the presence of CS(2) yielded the zwitterionic CS(2) adducts of the carbene, implicating the intermediacy of the free carbene in these reactions. PMID- 15760241 TI - Diametrically disubstituted cyclam derivative having Hg2+-selective fluoroionophoric behaviors. AB - [reaction: see text] A new fluoroionophore has been synthesized by appending two signaling pyrenylacetamide subunits on the binding motif of 1,8-dimethylcyclam. The designed compound exhibited highly selective and sensitive fluoroionophoric behavior toward Hg(2+) ions of excimer emission in aqueous dioxane (dioxane/H(2)O = 1:9, v/v) solution with a detection limit of 1.3 x 10(-)(6) M. The "ON-OFF" type signaling behavior of the fluoroionophore is due to the metal ion induced conformational changes from folded to open-winged conformations by exploiting the two nearby appended pyrenyl fluorophores. PMID- 15760242 TI - Total synthesis of an immunosuppressive glycolipid, (2S,3S,4R)-1-O- (alpha-d galactosyl)-2- tetracosanoylamino-1,3,4-nonanetriol. AB - [reaction: see text] A practical and efficient total synthesis of (2S,3S,4R)-1-O (alpha-d-galactosyl)-2-tetracosanoylamino-1,3,4-nonanetriol, OCH 1b, a potential therapeutic candidate for Th1-mediated autoimmune diseases, is described. The synthesis incorporates direct alkylation onto epoxide 5 and stereospecific halide ion catalyzed alpha-glycosidation reaction. A key intermediate 10 was obtained in only eight steps and 37% overall yield from commercially available d-arabitol 2, and the total synthesis of 1b was accomplished in 12 steps and 19% overall yield. This method will enable the synthesis of a variety of phytosphingolipids, especially that with the shorter sphingosine side chain than 1a, in a highly stereoselective manner. PMID- 15760243 TI - RuCl3/CeCl3/NaIO4: a new bimetallic oxidation system for the mild and efficient dihydroxylation of unreactive olefins. AB - [reaction: see text] The catalytic dihydroxylation of olefins represents a unique synthetic tool for the generation of two C,O-bonds with defined relative configuration. Whereas OsO(4) has been established as a very general dihydroxylation catalyst within the past 30 years, the less expensive and toxic isoelectronic RuO(4) has found only limited use for this type of oxygen-transfer reaction. High catalyst loading and undesired side reactions were severe drawbacks in RuO(4)-catalyzed oxidations of C,C-double bonds. Recently, we were able to improve the RuO(4)-catalyzed dihydroxylation by addition of Bronsted acids to the reaction mixture. This protocol proved to be of general applicability, however, certain limitations were observed. To address these problematic functional groups a new Lewis acid accelerated oxidation was developed. The use of only 10 mol % of CeCl(3) allowed a further decrease in the catalyst concentration down to 0.25 mol % while broadening the scope of the reaction. Silyl ethers and nitrogen containing functional groups are now tolerated in this optimized protocol. Furthermore, competing scission reactions are supressed in the presence of Lewis acid allowing longer reaction times and the successful oxidation of electron-deficient tetrasubstituted double bonds that cannot be oxidized using known dihydroxylation protocols. PMID- 15760244 TI - Synthesis of diindeno-fused 4H-cyclopenta[def]phenanthren-4-ones and related compounds via benzannulated enediynyl propargylic alcohols. AB - [reaction: see text] Treatment of propargylic diols 5-7 with thionyl chloride promoted a cascade sequence of reactions leading to dichlorides 10-12 and, after reduction with tributyltin hydride, the diindeno-fused 4H cyclopenta[def]phenanthrenes 13-15 in a single operation. Hydrolysis of 13 and 14 furnished 4H-cyclopenta[def]phenanthren-4-ones 16 and 17, respectively. Air oxidation of an alkaline solution of dichloride 11 produced diketone 18. PMID- 15760245 TI - Enantioselective approaches to potential MetAP-2 reversible inhibitors. AB - [reaction: see text] Enantioselective deprotonation of 4-substituted cyclohexanones and highly stereoselective conjugate addition of higher order mixed cuprates were the key steps in a concise synthesis of fumagalone-related molecules. The origin of the (low) biological activity of the new compounds as compared to fumagalone is briefly discussed. PMID- 15760246 TI - Perinatal depression: prevalence, screening accuracy, and screening outcomes. PMID- 15760247 TI - History and current status of endovascular management for the extracranial carotid and supra-aortic vessels. AB - Few procedures in the history of medicine have been more controversial than carotid artery stenting (CAS) for the management of carotid artery occlusive disease. Introduced just as the randomized trials were establishing carotid endarterectomy as the gold standard for carotid interventions, CAS has finally reached the point in its development when dedicated stenting systems are being tested in randomized clinical trials. Assisted by the concomitant use of distal protection devices, CAS has shown equipoise with endarterectomy in terms of safety at 30 days. This review summarizes the completed and ongoing CAS trials and the applications of endovascular techniques in the supra-aortic vessels. PMID- 15760248 TI - Advances in vascular brachytherapy over the last 10 years: focus on femoropopliteal applications. AB - Restenosis and the need for repeated interventions after percutaneous transluminal angioplasty (PTA) remain major drawbacks limiting a widespread application of this minimally invasive technique in the femoropopliteal segment. During the last decade, vascular brachytherapy (BT) emerged as a promising, novel technology with the potential to reduce the rates of restenosis. Meanwhile, several randomized controlled trials have suggested beneficial short and midterm effects of BT in the femoropopliteal arteries. However, despite substantial advances, many questions remain regarding the utility of vascular BT and its dissemination as a practical tool to prevent restenosis. The risk of complications, such as late stent thrombosis, edge effect, catch-up late restenosis, and potential aneurysm formation, as well as the logistical issues associated with the use of this technology in the catheterization laboratory, are delaying the acceptance of BT for routine use. This article reviews the developments of BT for restenosis prevention during the past decade, focusing on implications for peripheral endovascular treatment. PMID- 15760249 TI - Percutaneous endovascular therapy of renal artery stenosis: technical and clinical developments in the past decade. AB - Renal artery stenosis may initiate or exacerbate arterial hypertension and/or renal insufficiency. During the last decade, technical improvements of diagnostic and interventional endovascular tools have led to more widespread use of endoluminal renal artery revascularization and broader indications for this type of therapy. Since the first renal artery angioplasties performed by Felix Mahler and Andreas Gruntzig in 1978, numerous single-center studies have documented the benefits of percutaneous renal revascularization. In the early 1990s, stent implantation was added to the interventionist's armamentarium for treating renal artery stenosis due to atherosclerosis or fibromuscular dysplasia. The meta analysis of 3 randomized studies comparing balloon angioplasty with best medical therapy found intervention to be beneficial for blood pressure control but not for preservation of renal function. Despite the absence of randomized studies, there is mounting evidence that stenting of hemodynamically relevant atherosclerotic renal artery stenosis has a positive impact on blood pressure control and renal function. This article summarizes the technical improvements in these endovascular tools during the last decade and gives an overview concerning their clinical impact on renal artery revascularization. PMID- 15760250 TI - The influence of endovascular techniques on our surgical practice: a 10-year view. PMID- 15760251 TI - Local endovascular delivery, gene therapy, and cell transplantation for peripheral arterial disease. AB - Advances in catheter technology, gene identification, and cell biology may provide novel treatment options for patients with peripheral arterial disease (PAD) who are not candidates for standard revascularization procedures. Animal studies and recent results in human beings suggest that transfer of growth factors or regulatory genes and transplantation of progenitor cells may provide novel therapy options by inducing therapeutic angiogenesis or by inhibiting restenosis. This review will discuss the development of a variety of catheters for localized endovascular delivery, as well as the various cellular and genetic strategies that exist to restore blood flow to ischemic tissue and to reduce neointimal hyperplasia. PMID- 15760252 TI - An update on endovascular therapy of the lower extremities. AB - Considerable advances have been made over the last decade in percutaneous technology for treatment of atherosclerotic diseases in the iliac, femoropopliteal, and distal tibioperoneal arteries. While treatment strategies are well defined in the iliofemoral segment, where angioplasty and stenting perform well in appropriately selected lesions, the search for a durable transcatheter therapy for femoropopliteal and distal occlusive disease continues. The spectrum of treatment alternatives to angioplasty ranges from transcatheter plaque excision to laser ablation, rotational atherectomy, cryoplasty, brachytherapy, and stenting. We review in this article the status of percutaneous endovascular techniques for the treatment of lower extremity vascular occlusive disease. PMID- 15760253 TI - Development of vascular biology over the past 10 years: heme oxygenase-1 in cardiovascular homeostasis. AB - The study of vascular biology has provided strong evidence for the role that free radical attack plays in the pathogenesis of cardiovascular diseases. The endothelial cell (EC) dysfunction that results from exposure to oxidative stresses, such as oxidized LDL, influences vascular cell gene expression, promoting smooth muscle cell (SMC) mitogenesis and apoptosis. These factors also play an important role in atherogenesis, which is attenuated by antioxidants. Thus, antioxidants are important to understanding the pathophysiology of cardiovascular diseases and to constructing an effective treatment strategy for these patients. Over the last decade, there has been a tremendous interest in the biology of heme oxygenase-1 (HO-1), which exhibits antioxidant effects in various forms of tissue injury. Moreover, the reaction is also the major source of carbon dioxide (CO) in the body, which is a physiologically important gaseous vasodilator that inhibits SMC proliferation. Thus, HO-1-derived products provide various mechanisms to maintain cardiovascular homeostasis. We review recent work on the cellular and molecular biological aspects of the HO/CO system in vascular pathophysiology. PMID- 15760254 TI - Vascular imaging: an unparalleled decade. AB - Vascular imaging techniques, such as catheter angiography, ultrasound, computed tomography (CT), and magnetic resonance (MR), have all undergone unprecedented innovation and incredible technological leaps in the last 10 years. Ultrasound, CT, and MR have progressed in acquisition speed, resolution, and accuracy to the point that they have now supplanted the former mainstay, invasive catheter-based angiography, despite the advent of digitized angiographic image recording. This review explores the advantages and shortcomings of each technique and how they have changed the diagnosis and assessment of the cardiovascular system for endovascular intervention. PMID- 15760255 TI - Advances in endovascular techniques to treat failing and failed hemodialysis access. AB - During the decade since JEVT was inaugurated, we have witnessed the growing application of endovascular techniques for arteriovenous (AV) access in parallel with the evolution of endovascular therapy for arterial pathology. To date, few if any technologies have compared with balloon angioplasty for treating venous anastomotic stenosis, the most common cause of access failure. Only one device, which incorporates the principles of access graft design and self-expanding stent technology, has been uniquely conceived for this pathology. The encapsulated polytetrafluoroethylene stent-graft has achieved reasonable preliminary results, but randomized data is forthcoming. Technology to clear the clot from a thrombosed graft continues to evolve, but will never be as cost-effective as simple balloon thrombectomy. However, the pressure placed on providers to perform all percutaneous interventions and move away from open techniques continues to fuel interest in this component of treatment. Finally, the pursuit of a completely percutaneous AV access continues. As with endovascular procedures in general, whether or not the procedure is cost-effective or time-consuming seems to take a back seat to the all-percutaneous approach that so many seem to converge upon. Moreover, as most autogenous fistulas and AV grafts can be created with minimal incisions under local anesthesia, the pursuit of a completely percutaneous access system seems more like an academic exercise than a practical application of technology. We must try and avoid the tendency to "minimize invasiveness" with technology that is maximally intensive (and expensive), such as limiting ourselves to only percutaneous methods. Given the increasing pressure to have an all autogenous access program, current techniques that apply well in prosthetic grafts will need to be modified to accommodate the different biology of a native fistula. Clearly, the enlarging end-stage renal disease population will continue to provide endovascular specialists with clinically challenging problems requiring new and revolutionary technology. PMID- 15760257 TI - Ten years of advances in neuroendovascular procedures. PMID- 15760256 TI - A history of thrombolytic therapy. AB - Thrombolytic therapy has been available for the last 5 decades, but the modern era of thrombolysis began in the early 1990s, with the execution of 3 multicenter trials designed to compare this potentially less invasive therapy to the then standard of care for acute limb ischemia, open surgical revascularization. Even with the development of several bio-engineered lytic agents, the major risk of thrombolytic therapy continues to be bleeding complications. Nevertheless, data exist to suggest that thrombolysis should be considered as an adjunct to open surgery, percutaneous interventions, or, occasionally, as sole therapy for acute vascular occlusion. This review summarizes the developmental milestones in the history of thrombolysis and reviews data supporting its use in acute arterial occlusions. PMID- 15760258 TI - Ten years of advancements in interventional cardiology. PMID- 15760259 TI - Reflections on the past 10 years: an interventional radiologist's perspective. PMID- 15760261 TI - Intravascular stents in the last and the next 10 years. AB - The first balloon-expandable coronary stent was approved "for the prevention of restenosis" in 1994, the same year that the Journal of Endovascular Therapy was inaugurated. Since then, the development of the stent has paralleled the evolution of endovascular intervention as a new specialty. Innovators have pushed to explore new and varied stent applications outside the coronary arteries. Carotid stenting, transjugular intrahepatic portocaval shunts, and covered stents are a few of these new applications that have now become commonplace. Dozens of stent designs and several new materials have been tested to solve the problem of in-stent restenosis, but it is the drug-eluting stent (DES) that has emerged as the most promising, at least in the coronary arteries. However, the benefits of DES technology are not likely to be effective in the more pervasive forms of in stent restenosis, such as encountered in the femoropopliteal segment. In the future, technologies aimed at stimulating rather than inhibiting tissue response to an implant may be part of the next wave of developments, as we take aim against the poor and/or slow tissue incorporation that manifests as leaks and dislodgement. In the superficial femoral artery, for example, mechanical stresses that cause fractures and dislocations may be addressed by using a very flexible endovascular device with a tissue-friendly inner surface that promotes rapid stent endothelialization to counter the biological effects of motion and microtrauma. The rapidly developing fields of nanotechnology, microelectronics, and advanced materials technology will enable the surface engineer to design molecular-specific surfaces for a new generation of vascular devices. Interactive implantable or injectable microdevices aimed at providing specific information upon demand from an external source will revolutionize disease prevention, as emphasis shifts toward monitoring cardiovascular risk exposure. There is no doubt that during the next 10 years, we will witness impressive technological progress in the field of cardiovascular implantable devices. PMID- 15760262 TI - Update on endovascular treatment of cerebrovascular diseases. AB - Several developments in endovascular technology have greatly expanded the application of these techniques to treat extra- and intracranial cerebrovascular diseases. This review explores the indications, techniques, and clinical results for endovascular treatment of ischemic stroke and intracranial stenoses, aneurysms, and arteriovenous malformations. PMID- 15760260 TI - The top 12 advances in vascular medicine. AB - In the past decade, impressive strides have been made in the diagnosis and management of atherosclerotic, aneurysmal, and thromboembolic diseases, thanks in large part to the explosive growth in both vascular biology and clinical vascular medicine. We review what we consider to be the top 12 advances in this field: the discovery of nitric oxide, the metabolic syndrome, new thrombophilic disorders, therapeutic angiogenesis, endoluminal treatment of chronic venous disease, and a variety of drugs, including sildenafil, cilostazol, low-molecular-weight heparins, oral direct thrombin inhibitors, clopidogrel, statins, and angiotensin converting enzyme inhibitors and angiotensin-receptor blocking agents. PMID- 15760263 TI - Management of abdominal aortic aneurysm: a decade of progress. AB - Since the world was first introduced to the concept of endovascular aneurysm repair by Parodi's landmark procedures in 1990, stent-grafts have assumed a prominent role in the management of abdominal aortic aneurysm. Most modern systems are trackable, accurate, and secure. The resulting endovascular procedure is safe, durable, effective, and versatile. Perhaps the most significant increment in the applicability of the endovascular technique was achieved by the development of bifurcated stent-grafts, which dispensed with inadequate distal aortic implantation sites. Additional branches and fenestrations now permit endovascular repair in cases of thoracoabdominal, pararenal, juxtarenal, and bilateral iliac aneurysms. These advances in device performance have been accompanied by a rapid dissemination of necessary skills, leading to the development of a new superspecialty of vascular therapy, with elements of vascular surgery, interventional radiology, and interventional cardiology. PMID- 15760264 TI - The excimer laser: science fiction fantasy or practical tool? AB - Nearly 20 years ago, in vitro experiments left no doubt about the fact that laser light can ablate atherosclerotic plaque. The initial enthusiastic results with the technology, particularly in coronary arteries, were followed by reports showing unacceptably high restenosis and complication rates. These poor results were due to the premature application of an underdeveloped technology, a lack of understanding of laser/tissue interaction, and the use of incorrect lasing techniques. Consequently, and without discrimination, all lasers were banned from the catheterization laboratories for nearly a decade. Technological enhancements of the excimer laser, combined with refined catheter lasing techniques, resulted in greater debulking of atherosclerotic material in long superficial femoral artery occlusions. These results triggered the application of the excimer laser technique as an atherectomy tool in more complex lesions below the knee. The multicenter Laser Atherectomy for Critical Ischemia study clearly demonstrated that the excimer laser technology resulted in limb salvage rates >90% in patients with critical limb ischemia (CLI). Furthermore, new clinical results indicate that the excimer laser is very effective in dissolving thrombotic obstructions, redirecting this technology to the coronary field. The results of the excimer laser in CLI validate the role of the cool laser in treating complex peripheral vascular disease. The results suggest a larger indication for this technology and support a more aggressive use of these interventional techniques in the treatment of this large patient cohort. However, all lasers are not equally effective in debulking atherosclerotic material. Only the athermic process associated with the excimer laser produces a safe and effective endovascular ablation of obstructive atherosclerotic and/or thrombotic material. The appropriate and safe utilization of the equipment and lasing techniques, combined with correct indications and patient selection, will contribute to the successful application of laser assisted atherectomy in complex peripheral and coronary artery obstructive disease. Unfortunately, little consistent scientific data has been generated to convince the interventional community of the usefulness of excimer laser ablation. PMID- 15760265 TI - Endografting of the aortic arch. AB - In recent years, endovascular repair with stent-grafts has made great advances as a minimally invasive alternative to conventional open surgery in the treatment of aortic aneurysm and dissection. Although many commercial endograft systems are now used worldwide for the treatment of these pathologies in the abdominal aorta, only a few dedicated stent-grafts have been developed for use in the thoracic aorta. However, these second-generation commercial endografts have almost identical specifications and performance profiles in terms of structure, function, and delivery mechanism as stent-graft systems employed in the abdominal aorta. Thus, endografts have been used in the thoracic aorta with little consideration to the morphological and hemodynamic characteristics specific to the aortic arch and the deployment techniques needed to navigate this curved region of the thoracic aorta. This review will survey the literature on aortic arch stent-graft repair and identify key elements critical to the successful design of an endograft to treat lesions in the aortic arch. PMID- 15760266 TI - Endograft technology: highlights of the past 10 years. AB - The past decade has seen the evolution of an exciting technology that has changed forever the treatment of aortic aneurysmal disease. From rather crude homemade stent-grafts constructed in the surgical suite to elegant commercially manufactured devices in a variety of configurations and sizes, the aortic endograft has experienced a meteoric rise in popularity to become a beneficial, minimally invasive therapy that can obviate the risk of rupture and death. There are now 3 approved endovascular devices on the market for infrarenal abdominal aortic aneurysm repair, and it is likely that additional and improved devices will become available in the future. This review revisits the developmental history of the aortic endograft, noting the ongoing refinements that have arisen from our experiences with the growing population of stent-graft patients. Although research continues to search for solutions to the problems of endoleak and migration, long-term results even with the earlier second and third generation devices are better than has been achieved with open surgical repair. PMID- 15760267 TI - A decade of thoracic endografting: planning the next 10 years... AB - Endovascular repair has emerged as a very important treatment modality in the management of a host of serious and relatively frequent thoracic aortic diseases. This minimally invasive approach is certain to revolutionize the entire field of thoracic aortic surgery in the near future. The technologies, however, can still be considered as a "work in progress." Future refinements, which should be available within the next 1 to 2 years, will address some of the most important needs as yet unmet today: lower profile (<22 F) delivery systems, greater device flexibility, no longitudinal metal bars, and precise deliverability and deployment characteristics that will be conducive to optimal success and safety, especially in the region of the aortic arch. PMID- 15760269 TI - Two distinct E3 ubiquitin ligases have complementary functions in the regulation of delta and serrate signaling in Drosophila. AB - Signaling by the Notch ligands Delta (Dl) and Serrate (Ser) regulates a wide variety of essential cell-fate decisions during animal development. Two distinct E3 ubiquitin ligases, Neuralized (Neur) and Mind bomb (Mib), have been shown to regulate Dl signaling in Drosophila melanogaster and Danio rerio, respectively. While the neur and mib genes are evolutionarily conserved, their respective roles in the context of a single organism have not yet been examined. We show here that the Drosophila mind bomb (D-mib) gene regulates a subset of Notch signaling events, including wing margin specification, leg segmentation, and vein determination, that are distinct from those events requiring neur activity. D-mib also modulates lateral inhibition, a neur- and Dl-dependent signaling event, suggesting that D-mib regulates Dl signaling. During wing development, expression of D-mib in dorsal cells appears to be necessary and sufficient for wing margin specification, indicating that D-mib also regulates Ser signaling. Moreover, the activity of the D-mib gene is required for the endocytosis of Ser in wing imaginal disc cells. Finally, ectopic expression of neur in D-mib mutant larvae rescues the wing D-mib phenotype, indicating that Neur can compensate for the lack of D-mib activity. We conclude that D-mib and Neur are two structurally distinct proteins that have similar molecular activities but distinct developmental functions in Drosophila. PMID- 15760271 TI - Molecular insights into human brain evolution. PMID- 15760270 TI - PGC-1alpha deficiency causes multi-system energy metabolic derangements: muscle dysfunction, abnormal weight control and hepatic steatosis. AB - The gene encoding the transcriptional coactivator peroxisome proliferator activated receptor-gamma coactivator-1alpha (PGC-1alpha) was targeted in mice. PGC-1alpha null (PGC-1alpha(-/-)) mice were viable. However, extensive phenotyping revealed multi-system abnormalities indicative of an abnormal energy metabolic phenotype. The postnatal growth of heart and slow-twitch skeletal muscle, organs with high mitochondrial energy demands, is blunted in PGC-1alpha( /-) mice. With age, the PGC-1alpha(-/-) mice develop abnormally increased body fat, a phenotype that is more severe in females. Mitochondrial number and respiratory capacity is diminished in slow-twitch skeletal muscle of PGC-1alpha( /-) mice, leading to reduced muscle performance and exercise capacity. PGC 1alpha(-/-) mice exhibit a modest diminution in cardiac function related largely to abnormal control of heart rate. The PGC-1alpha(-/-) mice were unable to maintain core body temperature following exposure to cold, consistent with an altered thermogenic response. Following short-term starvation, PGC-1alpha(-/-) mice develop hepatic steatosis due to a combination of reduced mitochondrial respiratory capacity and an increased expression of lipogenic genes. Surprisingly, PGC-1alpha(-/-) mice were less susceptible to diet-induced insulin resistance than wild-type controls. Lastly, vacuolar lesions were detected in the central nervous system of PGC-1alpha(-/-) mice. These results demonstrate that PGC-1alpha is necessary for appropriate adaptation to the metabolic and physiologic stressors of postnatal life. PMID- 15760273 TI - Chromosome cohesion: a cycle of holding together and falling apart. PMID- 15760272 TI - The immune epitope database and analysis resource: from vision to blueprint. PMID- 15760274 TI - Funding the way to open access. PMID- 15760275 TI - Microfauna-macrofauna interaction in the seafloor: lessons from the tubeworm. PMID- 15760276 TI - Expressing the big picture. PMID- 15760277 TI - Spectral properties of topical retinoids prevent DNA damage and apoptosis after acute UV-B exposure in hairless mice. AB - We showed in a recent study that topical retinyl palmitate prevented UV-B-induced DNA damage and erythema in humans. Given that retinyl palmitate is a precursor of retinoic acid, the biological form of vitamin A that acts through nuclear receptors, we wondered whether these protective effects toward UV-B exposure were either receptor dependent or linked to other properties of the retinoid molecule such as its spectral properties. We determined the epidermal retinoid profile induced by topical retinoic acid in hairless mice and analyzed its effect on markers of DNA photodamage (thymine dimers) and apoptosis following acute UV-B exposure; we compared these effects to those induced by other natural topical retinoids (retinaldehyde, retinol and retinyl palmitate) which do not directly activate the retinoid receptors. We then analyzed the direct action of these retinoids on UV-B-induced DNA damage and apoptosis in cultured A431 keratinocytes. Topical retinoic acid significantly decreased (approximately 50%) the number of apoptotic cells, as well as the formation of thymine dimers in the epidermis of mice exposed to acute UV-B. Interestingly, the other topical retinoids decreased apoptosis and DNA damage in a similar way. On the other hand, neither retinoic acid nor the other retinoids interfered with the apoptotic process in A431 keratinocytes exposed to UV-B, whereas DNA photodamage was slightly decreased. We conclude that the decrease of apoptotic cells in hairless mouse epidermis following topical retinoids and UV-B irradiation reflects a protection of the primary targets of UV-B (DNA) by a mechanism independent of the activation of retinoid nuclear receptors, rather than a direct inhibition of apoptosis. PMID- 15760278 TI - Light controls growth and development via a conserved pathway in the fungal kingdom. AB - Light inhibits mating and haploid fruiting of the human fungal pathogen Cryptococcus neoformans, but the mechanisms involved were unknown. Two genes controlling light responses were discovered through candidate gene and insertional mutagenesis approaches. Deletion of candidate genes encoding a predicted opsin or phytochrome had no effect on mating, while strains mutated in the white collar 1 homolog gene BWC1 mated equally well in the light or the dark. The predicted Bwc1 protein shares identity with Neurospora crassa WC-1, but lacks the zinc finger DNA binding domain. BWC1 regulates cell fusion and repression of hyphal development after fusion in response to blue light. In addition, bwc1 mutant strains are hypersensitive to ultraviolet light. To identify other components required for responses to light, a novel self-fertile haploid strain was created and subjected to Agrobacterium-mediated insertional mutagenesis. One UV-sensitive mutant that filaments equally well in the light and the dark was identified and found to have an insertion in the BWC2 gene, whose product is structurally similar to N. crassa WC-2. The C. neoformans Bwc1 and Bwc2 proteins interact in the yeast two-hybrid assay. Deletion of BWC1 or BWC2 reduces the virulence of C. neoformans in a murine model of infection; the Bwc1-Bwc2 system thus represents a novel protein complex that influences both development and virulence in a pathogenic fungus. These results demonstrate that a role for blue/UV light in controlling development is an ancient process that predates the divergence of the fungi into the ascomycete and basidiomycete phyla. PMID- 15760279 TI - A life course approach to chronic disease epidemiology. AB - A life course approach to chronic disease epidemiology uses a multidisciplinary framework to understand the importance of time and timing in associations between exposures and outcomes at the individual and population levels. Such an approach to chronic diseases is enriched by specification of the particular way that time and timing in relation to physical growth, reproduction, infection, social mobility, and behavioral transitions, etc., influence various adult chronic diseases in different ways, and more ambitiously, by how these temporal processes are interconnected and manifested in population-level disease trends. In this review, we discuss some historical background to life course epidemiology and theoretical models of life course processes, and we review some of the empirical evidence linking life course processes to coronary heart disease, hemorrhagic stroke, type II diabetes, breast cancer, and chronic obstructive pulmonary disease. We also underscore that a life course approach offers a way to conceptualize how underlying socio-environmental determinants of health, experienced at different life course stages, can differentially influence the development of chronic diseases, as mediated through proximal specific biological processes. PMID- 15760280 TI - Advances in cancer epidemiology: understanding causal mechanisms and the evidence for implementing interventions. AB - In a worldwide population of 6 billion, in the year 2000, approximately 10 million cancers were diagnosed, and there were an estimated 6.2 million cancer deaths. Whereas the universality of cancer incidence and mortality is established, the burden of cancer by type or organ site is distributed unequally between developing and industrialized nations. Populations in developing countries are disproportionately affected by cancers in which infectious agents are causal. Our review of advances in cancer epidemiology underscores the complexity of pathogenic mechanisms mediated by chronic inflammation, obesity, and gene-environment interactions as in tobacco and alcohol carcinogenesis. Ultimately, the implementation of effective cancer control interventions that will serve to alleviate the cancer burden must integrate basic and applied research in the behavioral, social, biomedical, and population sciences. PMID- 15760281 TI - Competing dietary claims for weight loss: finding the forest through truculent trees. AB - In response to an accelerating obesity pandemic, competing weight-loss diets have propagated; those touting carbohydrate restriction are currently most in vogue. Evidence that sustainable weight loss is enhanced by means other than caloric restriction, however, is lacking. Whereas short-term weight loss is consistently achieved by any dietary approach to the restriction of choice and thereby calories, lasting weight control is not. Competing dietary claims imply that fundamental knowledge of dietary pattern and human health is lacking; an extensive literature belies this notion. The same dietary and lifestyle pattern conducive to health promotion is consistently associated with weight control. A bird's eye view of the literature on diet and weight reveals a forest otherwise difficult to discern through the trees. Competing diet claims are diverting attention and resources from what is actually and urgently needed: a dedicated and concerted effort to make the basic dietary pattern known to support both health and weight control more accessible to all. PMID- 15760282 TI - Population disparities in asthma. AB - The prevalence of asthma in the United States is higher than in many other countries in the world. Asthma, the most common chronic disease of childhood in the United States, disproportionately burdens many socioeconomically disadvantaged urban communities. In this review we discuss hypotheses for between country disparities in asthma prevalence, including differences in "hygiene" (e.g., family size, use of day care, early-life respiratory infection exposures, endotoxin and other farm-related exposures, microbial colonization of the infant bowel, exposure to parasites, and exposure to large domestic animal sources of allergen), diet, traffic pollution, and cigarette smoking. We present data on socioeconomic and ethnic disparities in asthma prevalence and morbidity in the United States and discuss environmental factors contributing to asthma disparities (e.g., housing conditions, indoor environmental exposures including allergens, traffic air pollution, disparities in treatment and access to care, and cigarette smoking). We discuss environmental influences on somatic growth (low birth weight, prematurity, and obesity) and their relevance to asthma disparities. The relevance of the hygiene hypothesis to the U.S. urban situation is reviewed. Finally, we discuss community-level factors contributing to asthma disparities. PMID- 15760283 TI - The rise and fall of menopausal hormone therapy. AB - Clinical trials show that hormone therapy (HT) is an effective treatment for vasomotor symptoms and vaginal dryness. HT improves other symptoms including sleep and quality of life in women who have menopause symptoms. In the Women's Health Initiative controlled clinical trials, both estrogen therapy (ET) and estrogen plus progestin therapy (EPT) reduced fracture risk, neither reduced the risk of heart disease, and both increased the risk of stroke, deep vein thrombosis, and dementia. EPT, but not ET, increased breast cancer risk and reduced colon cancer risk. Differences between EPT and ET may reflect chance, baseline differences between the EPT and ET cohorts, or a progestin effect. Studies of younger women and lower HT doses with intermediate endpoints are beginning. PMID- 15760284 TI - Advances in risk assessment and communication. AB - Risk analysis continues to evolve. There is increasing depth and breadth to each component of the four-step risk-assessment paradigm of hazard identification, dose-response analysis, exposure assessment, and risk characterization. Basic conceptual approaches to understanding how people perceive risk are being tested against a growing body of empirical observations, many involving stakeholders. Emerging ideas such as the precautionary principle have provided challenges that have led to a rethinking of the role of risk assessment in environmental health. Newer problems, such as intergenerational issues posed by long-lasting radiation pollution, environmental justice, and the assessment and communication of risks related to terrorism, have spurred innovative approaches to risk analysis. PMID- 15760285 TI - EMF and health. AB - Electric and magnetic fields are ubiquitous in the modern society, and concerns have been expressed regarding possible adverse effects of these exposures. This review covers epidemiologic research on health effects of exposures to static, extremely low-frequency (ELF), and radio frequency (RF) fields. Research on ELF fields has been performed for more than two decades, and the methodology and quality of studies have improved over time. Studies have consistently shown increased risk for childhood leukemia associated with ELF magnetic fields, whereas ELF fields most likely are not a risk factor for breast cancer and cardiovascular disease. There are still inadequate data for other outcomes. More recently, focus has shifted toward RF exposures from mobile telephony. There are no persuasive data suggesting a health risk, but this research field is still immature with regard to the quantity and quality of available data. This technology is constantly changing and there is a need for continued research on this issue. Almost no epidemiologic data are available for static fields. PMID- 15760286 TI - The public health impact of prion diseases. AB - Several prion disease-related human health risks from an exogenous source can be identified in the United States, including the iatrogenic transmission of Creutzfeldt-Jakob disease (CJD), the possible occurrence of variant CJD (vCJD), and potential zoonotic transmission of chronic wasting disease (CWD). Although cross-species transmission of prion diseases seems to be limited by an apparent "species barrier," the occurrence of bovine spongiform encephalopathy (BSE) and its transmission to humans indicate that animal prion diseases can pose a significant public health risk. Recent reports of secondary person-to-person spread of vCJD via blood products and detection of vCJD transmission in a patient heterozygous at codon 129 further illustrate the potential public health impacts of BSE. PMID- 15760287 TI - Water and bioterrorism: preparing for the potential threat to U.S. water supplies and public health. AB - Water supplies and water distribution systems represent potential targets for terrorist activity in the United States because of the critical need for water in every sector of our industrialized society. Even short-term disruption of water service can significantly impact a community, and intentional contamination of a municipal water system as part of a terrorist attack could lead to serious medical, public health, and economic consequences. Most practicing physicians and public health professionals in the United States have received limited training in the recognition and evaluation of waterborne disease from either natural or intentional contamination of water. Therefore, they are poorly prepared to detect water-related disease resulting from intentional contamination and may not be adequately trained to respond appropriately to a terrorist assault on water. The purpose of this review is to address this critical information gap and present relevant epidemiologic and clinical information for public health and medical practitioners who may be faced with addressing the recognition, management, and prevention of water terrorism in their communities. PMID- 15760288 TI - Economic causes and consequences of obesity. AB - Obesity is not only a health but also an economic phenomenon. This chapter (a) examines underlying economic causes, such as technological advancements, behind the obesity epidemic; (b) describes economic consequences of obesity, including increasing obesity-related medical expenditures; and (c) discusses the role of government in combating the obesity epidemic. Because of the high costs of obesity, and the fact that the majority of these costs are financed by taxpayers, there is a clear motivation for government to try to reduce these costs. However, because obesity may result from poor information and addictive behavior and/or as a result of living in an increasingly obesogenic environment, interventions will need to be multifaceted to ensure the best chance of success. PMID- 15760290 TI - New microbiology tools for public health and their implications. AB - The realm of diagnostic assays for detection of acute infections is rapidly changing from antibody detection to pathogen detection, from clinical laboratory based to point-of-care based, from single analyte detection to multiple analyte detection, and is more focused on detection using less invasive approaches for collecting biological samples. New assays are typically more sensitive than are conventional assays and have the capability of providing more information that characterizes the pathogen or the host response to the pathogen. From a public health perspective, the advent of molecular epidemiology, which allows tracking of pathogens based on unique genetic sequences or antigenic properties, has revolutionized how epidemiologists investigate and evaluate epidemics and assess endemic diseases. In addition, the use of point-of-care (POC) devices can impact the detection and surveillance of infections and will enhance our ability to accurately identify the causes of illnesses. PMID- 15760289 TI - Magnitude of alcohol-related mortality and morbidity among U.S. college students ages 18-24: changes from 1998 to 2001. AB - Integrating data from the National Highway Traffic Safety Administration, the Centers for Disease Control and Prevention, national coroner studies, census and college enrollment data for 18-24-year-olds, the National Household Survey on Drug Abuse, and the Harvard College Alcohol Survey, we calculated the alcohol related unintentional injury deaths and other health problems among college students ages 18-24 in 1998 and 2001. Among college students ages 18-24 from 1998 to 2001, alcohol-related unintentional injury deaths increased from nearly 1600 to more than 1700, an increase of 6% per college population. The proportion of 18 24-year-old college students who reported driving under the influence of alcohol increased from 26.5% to 31.4%, an increase from 2.3 million students to 2.8 million. During both years more than 500,000 students were unintentionally injured because of drinking and more than 600,000 were hit/assaulted by another drinking student. Greater enforcement of the legal drinking age of 21 and zero tolerance laws, increases in alcohol taxes, and wider implementation of screening and counseling programs and comprehensive community interventions can reduce college drinking and associated harm to students and others. PMID- 15760291 TI - The public health infrastructure and our nation's health. AB - Threats to Americans' health-including chronic disease, emerging infectious disease, and bioterrorism-are present and growing, and the public health system is responsible for addressing these challenges. Public health systems in the United States are built on an infrastructure of workforce, information systems, and organizational capacity; in each of these areas, however, serious deficits have been well documented. Here we draw on two 2003 Institute of Medicine reports and present evidence for current threats and the weakness of our public health infrastructure. We describe major initiatives to systematically assess, invest in, rebuild, and evaluate workforce competency, information systems, and organizational capacity through public policy making, practical initiatives, and practice-oriented research. These initiatives are based on applied science and a shared federal-state approach to public accountability. We conclude that a newly strengthened public health infrastructure must be sustained in the future through a balancing of the values inherent in the federal system. PMID- 15760292 TI - Social marketing in public health. AB - Social marketing, the use of marketing to design and implement programs to promote socially beneficial behavior change, has grown in popularity and usage within the public health community. Despite this growth, many public health professionals have an incomplete understanding of the field. To advance current knowledge, we provide a practical definition and discuss the conceptual underpinnings of social marketing. We then describe several case studies to illustrate social marketing's application in public health and discuss challenges that inhibit the effective and efficient use of social marketing in public health. Finally, we reflect on future developments in the field. Our aim is practical: to enhance public health professionals' knowledge of the key elements of social marketing and how social marketing may be used to plan public health interventions. PMID- 15760293 TI - Urban health: evidence, challenges, and directions. AB - Urbanization is one of the most important demographic shifts worldwide during the past century and represents a substantial change from how most of the world's population has lived for the past several thousand years. The study of urban health considers how characteristics of the urban environment may affect population health. This paper reviews the empirical research assessing urban living's impact on population health and our rationale for considering the study of urban health as a distinct field of inquiry. The key factors affecting health in cities can be considered within three broad themes: the physical environment, the social environment, and access to health and social services. The methodologic and conceptual challenges facing the study of urban health, arising both from the limitations of the research to date and from the complexities inherent in assessing the relations among complex urban systems, disease causation, and health are discussed. PMID- 15760295 TI - Adolescent resilience: a framework for understanding healthy development in the face of risk. AB - Adolescent resilience research differs from risk research by focusing on the assets and resources that enable some adolescents to overcome the negative effects of risk exposure. We discuss three models of resilience-the compensatory, protective, and challenge models-and describe how resilience differs from related concepts. We describe issues and limitations related to resilience and provide an overview of recent resilience research related to adolescent substance use, violent behavior, and sexual risk behavior. We then discuss implications that resilience research has for intervention and describe some resilience-based interventions. PMID- 15760296 TI - Declining rates of physical activity in the United States: what are the contributors? AB - This review describes current patterns and long-term trends (up to 50 years when possible) related to (a) physical activity, (b) employment and occupation, (c) travel behavior, (d) land use, and (e) related behaviors (e.g., television watching). On the basis of available data, the following trends were observed according to type of physical activity: relatively stable or slightly increasing levels of leisure-time physical activity, declining work-related activity, declining transportation activity, declining activity in the home, and increasing sedentary activity. These result in an overall trend of declining total physical activity. Large differences were noted in the rates of walking for transportation across metropolitan statistical areas. A strong linear increase existed in vehicle miles traveled per person over the past half century, coupled with a strong and consistent trend toward Americans living in suburbs. Although it is difficult to precisely quantify owing to the lack of long-term data, it is apparent that a combination of changes to the built environment and increases in the proportion of the population engaging in sedentary activities put the majority of the American population at high risk of physical inactivity. PMID- 15760294 TI - Acculturation and Latino health in the United States: a review of the literature and its sociopolitical context. AB - This chapter provides an overview of the concept of acculturation and reviews existing evidence about the possible relationships between acculturation and selected health and behavioral outcomes among Latinos. The effect of acculturation on Latino health is complex and not well understood. In certain areas-substance abuse, dietary practices, and birth outcomes-there is evidence that acculturation has a negative effect and that it is associated with worse health outcomes, behaviors, or perceptions. In others-health care use and self perceptions of health-the effect is mostly in the positive direction. Although the literature, to date, on acculturation lacks some breadth and methodological rigor, the public health significance of findings in areas in which there is enough evidence justifies public health action. We conclude with a set of general recommendations in two areas-public health practice and research-targeted to public health personnel in academia, community-based settings, and government agencies. PMID- 15760297 TI - Primary prevention of diabetes: what can be done and how much can be prevented? AB - Although it is widely believed that type 2 diabetes mellitus is the result of a complex interplay between genetic and environmental factors, compelling evidence from epidemiologic studies indicates that the current worldwide diabetes epidemic is largely due to changes in diet and lifestyle. Prospective cohort studies and randomized clinical trials have demonstrated that type 2 diabetes can be prevented largely through moderate diet and lifestyle modifications. Excess adiposity is the most important risk factor for diabetes, and thus, maintaining a healthy body weight and avoiding weight gain during adulthood is the cornerstone of diabetes prevention. Increasing physical activity and reducing sedentary behaviors such as prolonged TV watching are important both for maintaining body weight and improving insulin sensitivity. There is increasing evidence that the quality of fat and carbohydrate plays a more important role than does the quantity, and thus, public health strategies should emphasize replacing saturated and trans fats with unsaturated fats and replacing refined grain products with whole grains. Recent studies have also suggested a potential role for coffee, dairy, nuts, magnesium, and calcium in preventing diabetes. Overall, a healthy diet, together with regular physical activity, maintenance of a healthy weight, moderate alcohol consumption, and avoidance of sedentary behaviors and smoking, could nearly eliminate type 2 diabetes. However, there is still a wide gap between what we know and what we practice in the field of public health; how to narrow that gap remains a major public health challenge. PMID- 15760298 TI - Psychosocial factors and cardiovascular diseases. AB - Rapidly accruing evidence from a diversity of disciplines supports the hypothesis that psychosocial factors are related to morbidity and mortality due to cardiovascular diseases. We review relevant literature on (a) negative emotional states, including depression, anger and hostility, and anxiety; (b) chronic and acute psychosocial stressors; and (c) social ties, social support, and social conflict. All three of these psychosocial domains have been significantly associated with increased risk of cardiovascular morbidity and mortality. We also discuss critical pathophysiological mechanisms and pathways that likely operate in a synergistic and integrative way to promote atherogenesis and related clinical manifestations. We conclude by discussing some of the important challenges and opportunities for future investigations. PMID- 15760299 TI - Abortion in the United States. AB - Abortion is an extremely safe and common medical procedure. In the United States, over one million women had an abortion in the year 2000. Advances in early abortion techniques have helped to increase the proportion of early procedures, the safest type. Abortion rates have been declining since the early nineties among adults and adolescents, but rates among poor, minority women remain high. State restrictions to abortion have a larger impact on poor women and young women. Restrictions and regulations have also resulted in the concentration of abortion services in specialized clinics. These clinics are subject to harassment. The expansion of abortion services to more types of providers could increase access, as well as integrate abortion into women's health care. PMID- 15760300 TI - Patient perceptions of the quality of health services. AB - As calls are made for a more patient-centered health care system, it becomes critical to define and measure patient perceptions of health care quality and to understand more fully what drives those perceptions. This chapter identifies conceptual and methodological issues that make this task difficult, including the confusion between patient perceptions and patient satisfaction and the difficulty of determining whether systematic variations in patient perceptions should be attributed to differences in expectations or actual experiences. We propose a conceptual model to help unravel these knotty issues; review qualitative studies that report directly from patients on how they define quality; provide an overview of how health plans, hospitals, physicians, and health care in general are currently viewed by patients; assess whether and how patient health status and demographic characteristics relate to perceptions of health care quality; and identify where further, or more appropriately designed, research is needed. Our aim is to find out what patients want, need and experience in health care, not what professionals (however well-motivated) believe they need or get. PMID- 15760301 TI - Toward a system of cancer screening in the United States: trends and opportunities. AB - The hard work of public health officials, physicians, and disease advocacy groups to educate Americans about the importance of early detection has resulted in uptake of screening tests at levels equivalent to or higher than in countries with organized cancer screening programs. However, the societal costs of high screening rates are larger in the United States than in other countries, including higher prices for screening, more unnecessary testing, and inefficiencies in delivery, especially in small practices. Further, screening rates are not evenly distributed across population groups, and the national expenditure on clinical and community research to promote cancer screening among individuals has not been matched by research efforts that focus on policy or clinical systems to increase screening widely throughout the population. We identify opportunities for organizational change that improve access to use, improve quality, and promote cost effectiveness in cancer screening delivery. PMID- 15760302 TI - Impact of nicotine replacement therapy on smoking behavior. AB - This review summarizes evidence pertaining to the role of nicotine medications in smoking cessation and focuses particularly on evaluating evidence of the impact that nicotine replacement therapies (NRT) have had on altering population trends in smoking behavior. Accumulated evidence from controlled clinical trials has demonstrated that available forms of NRT (e.g., gum, transdermal patch, nasal spray, inhaler, and lozenge) increase quit rates compared with placebos by 50% 100%. However, despite the positive results from these studies, fewer than one in five smokers making a quit attempt do so with the benefit of NRT. Because not enough smokers are using NRT, the availability of NRT has not had a measurable impact on influencing population trends in smoking behavior. Among the factors contributing to the low utilization of nicotine medications are the inadequacies of the current dosage strengths and formulations of existing medications, smokers' perceptions of the high cost of the drugs, and concerns that many smokers have about safety and efficacy of nicotine medications. PMID- 15760303 TI - Activation of Mrc1, a mediator of the replication checkpoint, by telomere erosion. AB - BACKGROUND INFORMATION: In budding yeast, the loss of either telomere sequences (in telomerase-negative cells) or telomere capping (in mutants of two telomere end-protection proteins, Cdc13 and Yku) lead, by distinct pathways, to telomeric senescence. After DNA damage, activation of Rad53, which together with Chk1 represents a protein kinase central to all checkpoint pathways, normally requires Rad9, a checkpoint adaptor. RESULTS: We report that in telomerase-negative (tlc1Delta) cells, activation of Rad53, although diminished, could still take place in the absence of Rad9. In contrast, Rad9 was essential for Rad53 activation in cells that entered senescence in the presence of functional telomerase, namely in senescent cells bearing mutations in telomere end protection proteins (cdc13-1 yku70Delta). In telomerase-negative cells deleted for RAD9, Mrc1, another checkpoint adaptor previously implicated in the DNA replication checkpoint, mediated Rad53 activation. Rad9 and Rad53, as well as other DNA damage checkpoint proteins (Mec1, Mec3, Chk1 and Dun1), were required for complete DNA-damage-induced cell-cycle arrest after loss of telomerase function. However, unexpectedly, given the formation of an active Rad53-Mrc1 complex in tlc1Delta rad9Delta cells, Mrc1 did not mediate the cell-cycle arrest elicited by telomerase loss. Finally, we report that Rad9, Mrc1, Dun1 and Chk1 are activated by phosphorylation after telomerase inactivation. CONCLUSIONS: These results indicate that loss of telomere capping and loss of telomere sequences, both of which provoke telomeric senescence, are perceived as two distinct types of damages. In contrast with the Rad53-Rad9-mediated cell-cycle arrest that functions in a similar way in both types of telomeric senescence, activation of Rad53-Mrc1 might represent a specific response to telomerase inactivation and/or telomere shortening, the functional significance of which has yet to be uncovered. PMID- 15760305 TI - Effect of H2O2 on CCK-8-evoked changes in mitochondrial activity in isolated mouse pancreatic acinar cells. AB - BACKGROUND INFORMATION: This paper studies the effect of H(2)O(2) on mitochondrial responses evoked by CCK-8 (cholecystokinin 8) in mouse pancreatic acinar cells. Cytosolic ([Ca(2+)](c)) and mitochondrial ([Ca(2+)](m)) free calcium concentrations, mitochondrial inner membrane potential (psi(m)) and FAD autofluorescence were monitored using confocal laser scanning microscopy. RESULTS: CCK-8 induced an increase in [Ca(2+)](m) that slowly declined towards the prestimulation level. Depolarization of psi(m) that partially recovered, as well as increases in FAD autofluorescence, could also be observed in response to the hormone. Pretreatment of cells with 1 mM H(2)O(2) alone resulted in marked changes in mitochondrial parameters and, moreover, H(2)O(2) inhibited the CCK-8 evoked changes in [Ca(2+)](m), psi(m) and FAD autofluorescence. The results of the present study have demonstrated that CCK-8 can evoke marked changes in pancreatic acinar cell mitochondrial activity and that CCK-8-evoked responses are blocked by H(2)O(2). Additionally, H(2)O(2) releases Ca(2+) from intracellular stores and inhibits pancreatic acinar cell responses to CCK-8. CONCLUSION: The effects observed reflect an impairment of mitochondrial activity in the presence of H(2)O(2) that could represent some of its mechanisms of action to induce cellular damage leading to cell dysfunction and generation of pathologies. PMID- 15760304 TI - Mammalian cells stably overexpressing N-acylphosphatidylethanolamine-hydrolysing phospholipase D exhibit significantly decreased levels of N acylphosphatidylethanolamines. AB - In animal tissues, NAEs (N-acylethanolamines), including N arachidonoylethanolamine (anandamide), are primarily formed from their corresponding NAPEs (N-acylphosphatidylethanolamines) by a phosphodiesterase of the PLD (phospholipase D) type (NAPE-PLD). Recently, we cloned cDNAs of NAPE-PLD from mouse, rat and human [Okamoto, Morishita, Tsuboi, Tonai and Ueda (2004) J. Biol. Chem. 279, 5298-5305]. However, it remained unclear whether NAPE-PLD acts on endogenous NAPEs contained in the membrane of living cells. To address this question, we stably transfected two mammalian cell lines (HEK-293 and CHO-K1) with mouse NAPE-PLD cDNA, and investigated the endogenous levels and compositions of NAPEs and NAEs in these cells, compared with mock-transfected cells, with the aid of GC-MS. The overexpression of NAPE-PLD caused a decrease in the total amount of NAPEs by 50-90% with a 1.5-fold increase in the total amount of NAEs, suggesting that the recombinant NAPE-PLD utilizes endogenous NAPE as a substrate in the cell. Since the compositions of NAEs and NAPEs of NAPE-PLD-overexpressing cells and mock-transfected cells were very similar, the enzyme did not appear to discriminate among the N-acyl groups of endogenous NAPEs. These results confirm that overexpressed NAPE-PLD is capable of forming NAEs, including anandamide, in living cells. PMID- 15760307 TI - The Surgeon- a sculpture by Andor Meszaros. PMID- 15760308 TI - Therapeutic potential of stem cells in perinatal medicine. AB - Increasing evidence suggests that stem cells have tremendous potential to facilitate repair of damaged tissue and to exert protective influences that limit the extent of damage. Their inherent capacity to respond to signals generated by damaged tissue, migrate to these regions and either replace dead tissue or deliver protection by secretion of specific growth hormones and protective factors, suggests that they might have unrivalled therapeutic potential in perinatal medicine. A further potential of stem cells is their use in gene repair strategies for genetic disorders; an application which is exceedingly interesting from a perinatal perspective. Because of the relatively small size of infants and their capacity for future growth, stem cell therapy could be more successful in newborns than in older children or adults. In practical terms, the placenta, with its large reservoir of fetal blood, offers the ideal source of autologous stem cells. This affords the opportunity for stem cells to be collected and used, either directly ex vivo or after in vitro modulation, both for disorders in the neonatal period and for those arising later in life. The organs most affected from tissue damage in the neonatal period are the brain and the lung. So far, the most promising application of stem cells might be in the treatment of neurological injury. In this review we discuss recent research findings with adult stem cell therapy and their potential use in perinatal medicine. Furthermore, specific animal models suitable to explore the patho-physiological mechanisms of stem cell transplantation after neurological injury will be discussed. This review gives an overview of basic science findings and their possible role for clinical application with regards to the therapeutic potential of stem cells in perinatal medicine. Medline was searched for journal selection in peer-reviewed journals with high impact scores, which were relevant to this topic. All articles were in English and the search was not limited by publication year. However, the oldest publication was dated 1988 (reference 1). PMID- 15760309 TI - Impact of an early pregnancy problem service on patient care and Emergency Department presentations. AB - AIM: To examine the impact of a 'next day' outpatient clinic, the Early Pregnancy Problem Service, on patients presenting to the Emergency Department with pain or bleeding in the first trimester of pregnancy. This clinic was established in June 1996. METHODS: This was a retrospective study involving patients presenting to the Emergency Department with pain or bleeding in early pregnancy. Data was collected from the Emergency Department Information System and the medical records and then compared over the same 2-month periods in different years. These were in, 1996 (preclinic), 1997, 2000 and 2003. The primary outcome was length of stay in the Emergency Department for women with first trimester pain or bleeding that did not require hospital admission. Secondary outcomes were the proportion of such patients presenting and re-presenting to the Emergency Department. RESULTS: Following establishment of the clinic there was a significant reduction in the median length of stay in the Emergency Department for patients who were discharged (136 mins in 1996 vs 107 mins in 2003; P < 0.001). There were non significant reductions in the proportion of patients presenting to the Emergency Department (1.5% in 1996 vs 1.1% in 2003; P = 0.09) and the number re-presenting (16% in 1996 vs 7% in 2003; P = 0.15). CONCLUSION: Following the introduction of the Early Pregnancy Problem Service, women presenting with first trimester pain or bleeding who did not require emergency hospital admission spent significantly less time in the Emergency Department. PMID- 15760310 TI - Ultrasound localisation of non-palpable Implanon. AB - BACKGROUND: Localisation of the non-palpable Implanon is useful to confirm its presence or to assist the surgeon with its removal. Previous studies have suggested that almost all non-palpable Implanon implants can be located with ultrasound in phantoms. AIM: To determine whether diagnostic ultrasound is a reliable method for localising non-palpable Implanon implant in-vivo. METHODS: Ultrasound was carried out to localise the non-palpable Implanon implant. To ascertain the accuracy of the ultrasound findings, all patients were followed-up until the implant was removed or until it was proven absent by serial progesterone levels confirming ovulation or by negative etonogestrel levels. RESULTS: In 22 out of 23 patients the Implanon was correctly identified as present. The specificity is 95.7% (95%CI 79.0-99.2%), the positive predictive value is also 95.7%. In six out of seven patients the Implanon was correctly identified as absent. The sensitivity is 85.7% (95%CI 48.7-97.4%), the negative predictive value is also 85.7%. Four patients were excluded due to incomplete follow-up. CONCLUSION: Ultrasound is a reliable first line method for localising non-palpable Implanon. When the Implanon is localised on ultrasound, it is very likely present. When the Implanon is not seen on ultrasound, etonogestrel determination should be carried out to confirm its absence. PMID- 15760311 TI - Utility of fetal echocardiogram in high-risk patients. AB - AIM: Patients at high risk of fetal congenital heart disease are commonly referred for second trimester fetal echocardiogram. The objective of this study was to evaluate the utility of routine fetal echocardiogram in high-risk patients after the evaluation of the four-chamber/left ventricular outflow tract (LVOT) views during comprehensive second trimester anatomy ultrasound. METHODS: Second trimester comprehensive anatomy ultrasounds, which included a four-chamber/LVOT view, and subsequent fetal echocardiograms carried out at the Duke University Medical Center from January 1995 and July 2002 were reviewed. Those fetal echocardiograms carried out between 17 and 30 weeks gestation were included in the analysis. RESULTS: A total of 725 individual subjects met the inclusion criteria. Twenty-nine fetal echocardiograms were ultimately reported as abnormal. Of these, 19 had an abnormal four-chamber/LVOT view, four had a suboptimal view and six had a normal view. Of the six patients with a normal four-chamber/LVOT, all had been referred for echocardiogram based on the presence of other significant fetal anomalies noted at the time of second trimester anatomy ultrasound (3), documented aneuploidy (2), and significant fetal arrhythmia (1). CONCLUSION: Utility in carrying out fetal echocardiogram was seen in patients with an abnormal four-chamber/LVOT view, a suboptimal view in a high-risk patient, and the presence of other significant fetal abnormalities. Utility was not seen in patients with pre-existing diabetes mellitus. PMID- 15760313 TI - Efficacy of hyoscine-N-butyl bromide (Buscopan) suppositories as a cervical spasmolytic agent in labour. AB - OBJECTIVE: To evaluate the efficacy and safety of hyoscine-N-butyl bromide (Buscopan) suppositories in the active phase of labour, i.e. once the labour is established. METHODS: A non-randomised controlled prospective study was carried out on 200 women in labour. In the active phase of labour, at 3 cm or more cervical dilatation, 100 women were administered Buscopan via suppository and 100 women (control) were not given any drug. RESULTS: The duration of first stage of labour was 123.86 +/- 68.89 (mean(+)-standard deviation) minutes in the study group and 368.05 +/- 133.0 min in the control group. These differences were statistically significant. There were no differences in the duration of the second and third stages of labour. There was no increased incidence of operative deliveries in the study group. No adverse effects were noted on the mother or fetus. CONCLUSION: Hyoscine-N-butyl bromide (HBB, Buscopan) suppositories were highly effective in shortening the duration of the first stage of labour. PMID- 15760312 TI - A randomised trial of surgical, medical and expectant management of first trimester spontaneous miscarriage. AB - BACKGROUND: Medical management and expectant care have been considered possible alternatives to surgical evacuation of the uterus for first trimester spontaneous miscarriage in recent years. AIM: To compare the effectiveness and safety of medical and expectant management with surgical management for first trimester incomplete or inevitable miscarriage. METHODS: Forty women were recruited following diagnosis of incomplete or inevitable miscarriage, and randomised to surgical, medical or expectant care via an off-site, computerised enrollment system. The primary outcome was the effectiveness of medical (vaginal misoprostol) and expectant management relative to surgical evacuation, assessed at 10-14 days and 8 weeks post-recruitment. Infection, pain, bleeding, anxiety, depression, physical and emotional recovery were assessed also. Analysis was by intention-to-treat. RESULTS: Effectiveness at 8 weeks was lower for medical (80.0%) and expectant (78.6%) than for surgical management (100.0%). Two women in the medical group had confirmed infections. Bleeding lasted longer in the expectant group than in the surgical group. There were no significant differences in pain, physical recovery, anxiety or depression between the groups. 54.6%, 42.9% and 57.1% of the surgical, medical and expectant groups respectively would opt for the same treatment again. CONCLUSION: Expectant care appears to be sufficiently safe and effective to be offered as an option for women. Medical management might carry a higher risk of infection than surgical or expectant care. PMID- 15760314 TI - Use of umbilical-cerebral Doppler ratios in predicting fetal growth restriction in near-term fetuses. AB - OBJECTIVE: To compare the sensitivity and specificity of different umbilical cerebral ratios in the prediction and detection of fetal growth restriction in near-term fetuses when the umbilical arterial waveform is within normal. METHODS: A prospective cross-sectional observational study was carried out recruiting consecutive singleton pregnancies with clinically suspected fetal growth restriction after 34 weeks gestation. The umbilical-cerebral ratios were then calculated from the S/D, RI and PI values and correlated with immediate perinatal outcome. RESULTS: A total of 187 patients were recruited. Twelve cases had abnormal UA Doppler flow velocity waveform studies. Of the 175 with normal UA Doppler findings, 92 (53.1%) were confirmed to have fetal growth restriction (FGR) with birth weights below the tenth centile for gestation. The detection rate of FGR by ultrasound biometry was 96.7%. The mean umbilical artery S/D, RI and PI values were higher in the fetal growth restriction group, while the middle cerebral artery values were lower as compared to fetuses with no growth restriction. A small but significant difference was seen in the umbilical cerebral ratios of the different indices between the two groups. Receiver operator characteristic curves showed that there was little difference between the performances of the S/D, RI or PI ratios and all had limited power in predicting fetal growth restriction. CONCLUSION: In the presence of normal umbilical artery Doppler waveforms, umbilical-cerebral ratios have limited power to predict fetal growth restriction. PMID- 15760315 TI - Early medical abortion: a new regimen up to 49 days' gestation. AB - AIM: To evaluate a new regimen of mifepristone and misoprostol in early medical abortion up to 49 days of amenorrhoea. METHODS: One hundred healthy women requesting pregnancy termination up to 49 days gestation received 200 mg mifepristone followed by 800 microg misoprostol orally 24 h later. Statistical analysis was carried out by unpaired t-test. RESULTS: Ninety-six percent of patients aborted completely 4.3 h after they were given misoprostol. No significant side-effects were noted. The duration of bleeding correlated with gestational age (P-value 0.009). CONCLUSION: This new regimen of mifepristone misoprostol is effective in terminating early pregnancy, with shorter induction to abortion interval and greater acceptability. PMID- 15760316 TI - Postoperative analgesic requirements - total laparoscopic hysterectomy versus vaginal hysterectomy. AB - BACKGROUND: There is limited information available on the requirement for postoperative analgesic drugs in patients submitted to total laparoscopic hysterectomy (TLH) compared with patients undergoing vaginal hysterectomy (VH). AIM: To compare the postoperative analgesic requirements in patients who underwent a TLH with patients who had a VH. METHODS: Chart review of 53 patients who had TLH and 47 who had VH and were seen postoperatively by an acute pain management service in order to assess postoperative analgesic requirements. Patient controlled analgesia (PCA) was part of the standard protocol for postoperative pain management. Analgesic requirement was recorded as the mean doses of morphine and number of days that patients used non-steroidal anti inflammatory drugs (NSAIDs), oxycodone and tramadol. RESULTS: The requirement for total morphine was approximately half the dose in patients who had a TLH (10.8 +/ 12.6 mg) compared with patients who had a VH (19.4 +/- 21.9 mg) (P 0.017). The length of use of NSAIDs was significantly reduced in patients who had undergone a TLH (2.0 +/- 0.95 days) as compared with patients who had a VH (2.85 +/- 1.1 days) (P < 0.0001). CONCLUSIONS: Patients submitted to TLH require less postoperative analgesic drugs when compared with patients who had VH. Prospective randomised trials are warranted to compare analgesic requirements between patients submitted to TLH and VH. PMID- 15760317 TI - Routine investigations might be useful in pre-eclampsia, but not in gestational hypertension. AB - BACKGROUND: Women referred to secondary care with suspected pregnancy-induced hypertension (PIH) are commonly investigated with blood tests and cardiotocography (CTG), regardless of the clinical severity of their condition. Over-investigation might lead to inappropriate intervention. AIMS: To investigate how often abnormal blood test and CTG results occur in women with pre-eclampsia and gestational hypertension and in women who do not have pregnancy-induced hypertension. METHODS: Retrospective case note review of 526 consecutive women referred with suspected pregnancy-induced hypertension to a district hospital. The frequency of abnormal test results and the pregnancy outcomes were analysed according to clinical classification. RESULTS: 36% of women referred did not meet the clinical criteria for a diagnosis of pregnancy-induced hypertension. Abnormalities of platelet count and/or liver function were seen in 11% of women with pre-eclampsia and in less than 2% of women with gestational hypertension and in a similar proportion of women who did not have pregnancy-induced hypertension. Gestational hypertension was associated with increased induction and caesarean birth rates, but not with low birthweight or preterm delivery. Progression from gestational hypertension to pre-eclampsia was not predicted by blood test abnormalities. Support for the routine use of antenatal CTG was not found. CONCLUSIONS: A clinical diagnosis of pregnancy-induced hypertension should be confirmed before blood tests are ordered. The incidence of test abnormalities was only increased in pre-eclampsia and in gestational hypertension before term. CTG might only be of use in selected cases. PMID- 15760319 TI - Demographic characteristics and clinical course in infants with moderate or severe neonatal encephalopathy. AB - A chart review of 64 infants with moderate or severe neonatal encephalopathy showed that resuscitation was required for 61 (95%), respiratory support for 53 (83%) and anticonvulsants for 58 (91%). Death occurred in 2 (4%) infants with moderate encephalopathy and 12 (86%) with severe encephalopathy. In addition, subsequent neurodevelopment was abnormal in approximately a quarter of infants who survived after a moderate to severe encephalopathy. PMID- 15760318 TI - Reproductive outcome of women with unicornuate uterus. AB - This study was undertaken to examine the reproductive impact of unicornuate uterine abnormalities cases in a group of 571 women with a uterine anomaly. Hospital records of 1784 patients who presented to the infertility outpatient clinic with infertility, recurrent pregnancy loss, pain or acute abdomen during the study period between January 1991 and January 2001, were reviewed retrospectively. The reproductive performance of women with unicornuate uterus was poor, with a live birth rate of only 29.2%, prematurity rate of 44%, miscarriage rate of 29%, and an ectopic pregnancy rate of 4%. PMID- 15760320 TI - Effect of Implanon on insulin resistance in women with Polycystic Ovary Syndrome. AB - The majority of women with polycystic ovary syndrome (PCOS) have insulin resistance (IR) and have a high risk of developing diabetes. In this retrospective study of women with PCOS, Implanon use was associated with increased insulin resistance, as evidenced by a deterioration in the homeostasis model assessment (HOMA) assessment of insulin resistance, and increased fasting insulin levels. Implanon might increase the risk of diabetes and potentially cardiovascular disease in this already high-risk group. Further research is required to establish the ideal contraceptive preparation in PCOS. PMID- 15760321 TI - Posterior sling (infracoccygeal sacropexy): an alternative procedure for vaginal vault prolapse. AB - This study of 30 patients evaluated the effectiveness and safety of the posterior sling (infracoccygeal sacropexy) in the surgical treatment of vaginal vault prolapse. The patients were reevaluated 3 months, 6 months and yearly postoperatively. Coexisting preoperative symptoms of pelvic pain, urgency, nocturia and 'obstructed' micturition feeling were followed-up. There was remarkable improvement in vault prolapse and in coexisting symptoms. PMID- 15760322 TI - Comparison of pulse methotrexate and pulse dactinomycin in the treatment of low risk gestational trophoblastic neoplasia. AB - Methotrexate and dactinomycin are efficient drugs in the treatment of patients with low-risk gestational trophoblastic neoplasia (LRGTN). To compare the effectiveness of these two drugs in LRGTN, 46 patients were randomised to receive weekly intramuscular methotrexate at 30 mg/m(2) (n = 28) or intravenous dactinomycin at 1.25 mg/m(2) every 2 weeks (n = 18). Fourteen patients (50%) in the methotrexate group and 16 patients (89%) in the dactinomycin group achieved complete response. Greater patient convenience and a lower number of required visits make dactinomycin superior to other alternatives. PMID- 15760323 TI - Full term ovarian pregnancy: a case report. PMID- 15760324 TI - Recurrent intravenous leiomyomatosis with extension up the inferior vena cava. PMID- 15760325 TI - Squamous carcinoma arising in a grafted donor kidney presenting as high-grade atypia on routine Pap cytology. PMID- 15760326 TI - Growing teratoma syndrome in germ cell tumour of the ovary: a case report. PMID- 15760327 TI - Dedifferentiation in a mucinous tumour of the ovary. PMID- 15760328 TI - HELLP, eclampsia and posterior reversible encephalopathy in a young woman with streak ovary syndrome. PMID- 15760329 TI - Prenatal transfer of anticardiolipin antibodies associated with fatal neonatal aortic thrombosis. PMID- 15760331 TI - Administering late terminations in Western Australia. PMID- 15760333 TI - Re: Influence of body weight on plasma homocysteine in preeclampsia. PMID- 15760334 TI - Editorial: a model, MIA and Sox. PMID- 15760335 TI - Pigment pattern formation in the medaka embryo. AB - For the study of development of pigmentation, compared with mammalian models, fish offer the advantage of multiple chromatophore types and ready access to the developing embryo for observation and experimental manipulation. Compared with zebrafish embryos, medaka embryos have an additional unique chromatophore-type and superb properties for conditional mutation studies. The rich resources of medaka mutants, combined with data obtainable from other species, potentially offer information not otherwise readily available regarding chromatophore lineage. Here we summarize the embryonic development of normal medaka pigment pattern, based on observations using embryos of a panel of wild type and mutant fish. A more detailed description of development is available in the appendix of the on-line version of this paper (see Supplementary Material). We make some comparisons with zebrafish development to emphasize the increased power of both systems when utilized together. These two models will, in combination, be a powerful system for studies of the embryogenesis and evolution of pigmentation. PMID- 15760336 TI - Secrets to a healthy Sox life: lessons for melanocytes. AB - Sox proteins are transcriptional regulators with a high-mobility-group domain as sequence-specific DNA-binding domain. For function, they generally require other transcription factors as partner proteins. Sox proteins furthermore affect DNA topology and may shape the conformation of enhancer-bound multiprotein complexes as architectural proteins. Recent studies suggest that Sox proteins are tightly regulated in their expression by many signalling pathways, and that their transcriptional activity is subject to post-translational modification and sequestration mechanisms. Sox proteins are thus ideally suited to perform their many different functions as transcriptional regulators throughout mammalian development. Their unique properties also cause Sox proteins to escape detection in many standard transcription assays. In melanocytes, studies have so far focused on the Sox10 protein which functions both during melanocyte specification and at later times in the melanocyte lineage. During specification, Sox10 activates the Mitf gene as the key regulator of melanocyte development. At later stages, it ensures cell-type specific expression of melanocyte genes such as Dopachrome tautomerase. Both activities require cooperation with transcriptional partner proteins such as Pax-3, CREB and eventually Mitf. If predictions can be made from other cell lineages, further functions of Sox proteins in melanocytes may still lie ahead. PMID- 15760337 TI - Comparative study of eye defective worm 'menashi' and regenerating wild-type in planarian, Dugesia ryukyuensis. AB - Inbreeding of the sexualized planarian, Dugesia ryukyuensis, produces eye defective worms, menashi, in the F1 population. To study the effects of this mutation on the eye, we observed the eye-region of menashi using electron microscopy and compared it with the regenerating eye in wild-type worms. The intact eye of wild-type planarians consisted of a few pigment cells and a number of visual cells. Pigment cells containing spherically-shaped electron-dense melanosomes contacted each other and enclosed rhabdomes of visual cells. Rhabdomes had numerous tubular microvilli extending radially and touching the pigment cells. However, in menashi, various lengths of tubular microvilli were irregularly distributed near the pigment cells, which contained numerous electron lucent premelanosomes, and no adhesive structures were found between the pigment cells. The premelanosomes of menashi were equal in size to those seen after 2 days of regeneration in wild-type planarians and were similar in maturation to those found after 3 days of regeneration in wild-type planarian. These results suggest that menashi is defective in the mechanism(s) of developing pigment granules and constructing visual cells. These findings also suggest that pigment cells in menashi are defective in the mechanism(s) involved with cell adhesion. PMID- 15760338 TI - Inhibition of melanoma inhibitory activity (MIA) expression in melanoma cells leads to molecular and phenotypic changes. AB - The secreted protein melanoma inhibitory activity (MIA) is highly expressed in malignant melanoma but not in melanocytes and is associated with tumor progression in vivo. Here, we further investigated the functional role of MIA by inhibiting MIA expression of the human melanoma cell line HMB2 via stable antisense MIA cDNA transfection, and subsequent analysis of the cell clones. MIA deficient cell clones showed several changes in cell morphology and growth pattern. In monolayer and three-dimensional culture enhanced cell-cell contacts were formed. Furthermore, a re-induction of pigment synthesis in comparison with the amelanotic parental cell line HMB2 was observed. Molecular analyses revealed a re-expression of tyrosinase-related protein 1 (Trp-1) and tyrosinase in the MIA deficient cell clones necessary for melanin synthesis. In accordance, re expression of MIA in the MIA-deficient melanoma cell clones resulted in downregulation of Trp-1. To identify the molecular mechanisms of MIA regulating pigmentation, MITF and PAX3, two positive regulators of Trp-1 and tyrosinase transcription, and PIAS3, a negative regulator of MITF activity, were analyzed. Only in MIA-deficient cells, expression of PAX3 mRNA and MITF protein was found. In contrast, strong expression of PIAS3 was detected in HMB2 but not in the MIA deficient cells. To our knowledge this is the first report demonstrating a correlation between MIA expression and pigmentation and morphology of melanocytic cells. PMID- 15760339 TI - Principal expression of two mRNA isoforms (ABCB 5alpha and ABCB 5beta ) of the ATP-binding cassette transporter gene ABCB 5 in melanoma cells and melanocytes. AB - ATP-binding cassette (ABC) transporters play a pivotal role in physiology and pathology. We identified and cloned two novel mRNA isoforms (ABCB 5alpha and ABCB 5beta) of the ABC transporter ABCB 5 in human melanoma cells. The deduced ABCB 5alpha protein appears to be an altered splice variant containing only a putative ABC, whereas the ABCB 5beta isoform shares approximately 70% similarity with ABCB1 (MDR1) and has a deduced topological arrangement similar to that of the whole carboxyl terminal half of the ABCB1 gene product, P-glycoprotein, including an intact ABC. Northern blot, real-time PCR, and conventional RT-PCR were used to verify the expression profiles of ABCB 5alpha/beta. We found that the melanomas included among the NCI-60 panel of cell lines preferentially expressed both ABCB 5alpha and ABCB 5beta. However, ABCB 5alpha/beta expression was undetectable in two amelanotic melanomas (M14 and LOX-IMVI). The expression profile of ABCB 5alpha/beta in all of the other melanomas of the panel was confirmed both by RT PCR and by sequencing. Neither ABCB 5alpha nor ABCB 5beta expression was found in normal tissues such as liver, spleen, thymus, kidney, lung, colon, small intestines or placenta. ABCB 5alpha/beta mRNAs were also expressed in normal melanocytes and in retinal pigment epithelial cells, suggesting that ABCB 5alpha/beta expression is pigment cell-specific and might be involved in melanogenesis. Our findings indicate that expression of ABCB 5alpha/beta might possibly provide two novel molecular markers for differential diagnosis of melanomas and constitute potential molecular targets for therapy of melanomas. PMID- 15760340 TI - Human placental lipid induces melanogenesis through p38 MAPK in B16F10 mouse melanoma. AB - Melanogenesis is one of the characteristic functional activities of melanocyte/melanoma and is regulated via mitogen-activated protein kinase (MAPK) and Akt/protein kinase B (PKB) pathways. Placental total lipid fraction (PTLF), prepared from a hydroalcoholic extract of fresh term human placenta contains sphingolipids and was recently shown to stimulate melanogenesis via up-regulation of the key enzyme tyrosinase in B16F10 mouse melanoma cells. How such lipids mediate their effects on pigmentation and tyrosinase expression is a particularly important aspect of melanogenesis. To study the signaling that leads to tyrosinase expression, we have investigated the roles of the MAPK and Akt/PKB pathways in B16F10 melanoma cells in melanogenesis in response to PTLF. Treatment of cells with PTLF led to the time dependent phosphorylation of p38 MAPK. SB203580, a p38 MAPK inhibitor, completely blocked the PTLF-induced melanogenesis by inhibiting promoter activity and subsequent expression of tyrosinase. Phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002 a blocker of the Akt signaling pathway, or an inhibitor of MEK (MAPK/ERK Kinase), PD98059 when included along with PTLF was found to potentiate PTLF-induced phosphorylation of p38 MAPK together with tyrosinase expression and melanogenesis. The results suggest that the activation of p38 MAPK plays a crucial role in PTLF-induced B16F10 melanogenesis by up-regulating tyrosinase expression. PMID- 15760341 TI - Degradation of tyrosinase induced by phenylthiourea occurs following Golgi maturation. AB - Tyrosinase, the rate-limiting enzyme of melanin synthesis, is a di-copper metalloprotein that catalyzes the conversion of L-tyrosine to L-DOPAquinone. Phenylthiourea (PTU) is a well-known inhibitor of tyrosinase and melanin synthesis and is known to interact with sweet potato catechol oxidase, an enzyme possessing copper binding domain homology to tyrosinase. While PTU is frequently used to induce hypopigmentation in biological systems, little is known about its effects on tyrosinase and other melanogenic proteins. We have found that PTU induces degradation of tyrosinase but not of other melanogenic proteins including the tyrosinase-related metalloproteins tyrosinase-related protein (Tyrp)1 and Tyrp2. Using pulse-chase analysis coupled with glycosidase digestion, we observed that tyrosinase degradation occurs following complete maturation of the protein and that degradation was reversed by cysteine protease inhibitor E64 but not proteasome inhibitor N-acetyl-L-leucinyl-L-leucinyl-L-norleucinal. We conclude that PTU specifically induces tyrosinse degradation following Golgi maturation. Our data suggest that in addition to well-known ER-directed quality control, tyrosinase is also subject to post-Golgi quality control. PMID- 15760342 TI - Isolation, purification and physicochemical characterization of water-soluble Bacillus thuringiensis melanin. AB - Melanins are widely used in medicine, pharmacology, cosmetics and other fields. Although several technologies for the purification of water-insoluble dioxyphenylalanine (DOPA) melanins have been described, a source of water-soluble melanin is highly desirable. Here we describe an effective procedure for the isolation and purification of water-soluble melanin using the culture medium of Bacillus thuringiensis subsp. galleriae strain K1. Water-soluble melanin from this organism has an isoelectric point (pI=3.0-3.2) and was purified optimally by adsorbtion using the IA-1r resin and elution as a concentrated solution. The purified melanin obtained exhibited a similar infra-red absorbtion spectrum to synthetic melanin and contained quinolic and phenolic structures and an amino acid content of around 20% after acid hydrolysis. The molecular weight of the purified melanin determined by SDS-PAGE was 4 kDa and the electromagnetic spin resonance spectrum of the purified microbial melanin was a slightly asymmetric singlet without hyperfine structure with about 7 Gauss width of the line between points of the maximum incline and g=2.006. The concentration of paramagnetic centers in melanin is 0.21x10(18) spin/g. The results obtained provide a rapid, simple and inexpensive method for the large scale purification of water soluble melanin that may have widespread applications. PMID- 15760344 TI - A Tyrosinase missense mutation causes albinism in the Wistar rat. AB - Tyrosinase serves as a key enzyme in the synthesis of melanin. In humans mutations in the TYR gene are associated with type 1 oculocutaneous albinism (OCA1) that leads to reduced or absent pigmentation of skin, hair and eye. Various mutations causing OCA in man, mouse, rabbit and cattle have been identified throughout the Tyrosinase gene including nonsense, missense, frameshift and splice site alterations. Here we report a missense substitution at codon R299H in exon 2 of the Tyr gene in the albino Wistar rat. As this very exchange has already been described in OCA patients, our findings reinforce the significance of this region for normal catalytic activity of tyrosinase protein. PMID- 15760343 TI - Hemocyanin in the exoskeleton of crustaceans: enzymatic properties and immunolocalization. AB - We investigated the enzymatic properties and immunohistochemical localization of cuticular hemocyanin, a known oxygen transporter in the prawn Penaeus japonicus. The molecular weight of hemocyanin purified from the cuticle was estimated to be 67-77 k using SDS-PAGE, and the purified protein was effectively converted into a phenoloxidase-like enzyme by an SDS-treatment. The activated enzyme catalyzed the o-hydroxylation of monophenols and the oxidation of o-diphenols and was inhibited by typical inhibitors of phenoloxidase. These characteristics were nearly identical to the enzymatic properties of hemolymph hemocyanin. Immunological detection showed a diffuse distribution of hemocyanin over the exocuticle and endocuticle, and a higher signal level was observed in the latter. Based on these results, roles of hemocyanin in various physiological processes such as immune response and sclerotization of the cuticle were discussed. PMID- 15760345 TI - An emerging focus on plant ecological development. PMID- 15760346 TI - Ancient and recent polyploidy in angiosperms. PMID- 15760347 TI - The control of leaf development. AB - The formation of a leaf is a basic aspect of plant development. This review provides an overview of our present understanding of the process from initiation to the final form of the leaf. Molecular genetic and cell biology approaches have yielded significant advances in this area, adding not only to our knowledge of leaf development but also to fundamental principles in plant biology. These principles will be highlighted, as well as areas where our understanding is still incomplete, in particular the problem of coordinating the multifaceted steps involved in the generation of the leaf structure. PMID- 15760348 TI - The evolution of Ca2+ signalling in photosynthetic eukaryotes. AB - It is likely that cytosolic Ca2+ elevations have played a part in eukaryotic signal transduction for about the last 2 Gyr, being mediated by a group of molecules which are collectively known as the [Ca2+]cyt signalling toolkit. Different eukaryotes often display strikingly similar [Ca2+]cyt signalling elevations, which may reflect conservation of toolkit components (homology) or similar constraints acting on different toolkits (homoplasy). Certain toolkit components, which are presumably ancestral, are shared by plants and animals, but some components are unique to photosynthetic organisms. We propose that the structure of modern plant [Ca2+]cyt signalling toolkits may be explained by their modular adaptation from earlier pathways. PMID- 15760349 TI - Plant eco-devo: the potential of poplar as a model organism. AB - Ecological developmental genetics is the study of how ecologically significant traits originate in the genome and how the allelic combinations responsible are maintained in populations and species. Plant development involves a continuous feedback between growth and environment and the success of individual genotype x environment interactions determines the passage of alleles to the next generation: the adaptive recursion. Outbreeding plants contain a large amount of genetic variation, mostly in the form of single nucleotide polymorphisms (SNPs). One of the challenges of eco-devo is to distinguish neutral SNPs from those with ecological consequences. The complete genome sequence of Populus trichocarpa Torr. & A. Gray will be a significant aid in this endeavour. Occurring from California to Alaska, this is the first ecologically 'keystone' species to be sequenced. It has a rich natural history and is an obligate outbreeder. The individual sequenced, Nisqually-1, appears to be heterozygous on average about every 100 bp over the c. 500 million bp of the genome. Overlaid on this within individual variation is some ecologically based between-individual genotypic variation evident across the distribution of the species. The synthesis of information from genomics and ecology is now in prospect. This 'ecomolecular synthesis' is likely to provide a rich insight into the genomic basis of plant adaptation. PMID- 15760350 TI - Constraints on the evolution of adaptive phenotypic plasticity in plants. AB - The high potential fitness benefit of phenotypic plasticity tempts us to expect phenotypic plasticity as a frequent adaptation to environmental heterogeneity. Examples of proven adaptive plasticity in plants, however, are scarce and most plastic responses actually may be 'passive' rather than adaptive. This suggests that frequently requirements for the evolution of adaptive plasticity are not met or that such evolution is impeded by constraints. Here we outline requirements and potential constraints for the evolution of adaptive phenotypic plasticity, identify open questions, and propose new research approaches. Important open questions concern the genetic background of plasticity, genetic variation in plasticity, selection for plasticity in natural habitats, and the nature and occurrence of costs and limits of plasticity. Especially promising tools to address these questions are selection gradient analysis, meta-analysis of studies on genotype-by-environment interactions, QTL analysis, cDNA-microarray scanning and quantitative PCR to quantify gene expression, and two-dimensional gel electrophoresis to quantify protein expression. Studying plasticity along the pathway from gene expression to the phenotype and its relationship with fitness will help us to better understand why adaptive plasticity is not more universal, and to more realistically predict the evolution of plastic responses to environmental change. PMID- 15760351 TI - Plant growth forms: an ecological and evolutionary perspective. AB - Trees, shrubs, lianas and herbs have widely different mechanical architectures, which can also vary phenotypically with the environment. This review investigates how environmental effects, particularly mechanical perturbation, can influence biomechanical development in self-supporting and climbing growth forms. The bifacial vascular cambium is discussed in terms of its significance to growth form variation, ecology and evolution among extant plants, and during its appearance and early evolution. A key aspect of this developmental innovation concerned its potential for architectural and mechanical variation in response to environmental effects as well as optimizing hydraulic supply before the appearance of laminate leaves. Growth form diversity and its importance to past and present ecosystems are discussed in relation to both evolutionary constraints and ecological factors such as climatic change and atmospheric CO2 concentrations. We discuss how widely ranging growth forms such as climbers show a large range of developmental and phenotypic variation that has much to offer in understanding how the environment can modify plant development, particularly in terms of the bifacial vascular cambium. The broad approach we propose would benefit a wide range of studies from research into wood development to long-term ecological censuses of today's potentially changing ecosystems. PMID- 15760352 TI - A modular concept of phenotypic plasticity in plants. AB - Based on empirical evidence from the literature we propose that, in nature, phenotypic plasticity in plants is usually expressed at a subindividual level. While reaction norms (i.e. the type and the degree of plant responses to environmental variation) are a property of genotypes, they are expressed at the level of modular subunits in most plants. We thus contend that phenotypic plasticity is not a whole-plant response, but a property of individual meristems, leaves, branches and roots, triggered by local environmental conditions. Communication and behavioural integration of interconnected modules can change the local responses in different ways: it may enhance or diminish local plastic effects, thereby increasing or decreasing the differences between integrated modules exposed to different conditions. Modular integration can also induce qualitatively different responses, which are not expressed if all modules experience the same conditions. We propose that the response of a plant to its environment is the sum of all modular responses to their local conditions plus all interaction effects that are due to integration. The local response rules to environmental variation, and the modular interaction rules may be seen as evolving traits targeted by natural selection. Following this notion, whole-plant reaction norms are an integrative by-product of modular plasticity, which has far reaching methodological, ecological and evolutionary implications. PMID- 15760353 TI - Niche construction through phenological plasticity: life history dynamics and ecological consequences. AB - The ability of an organism to alter the environment that it experiences has been termed 'niche construction'. Plants have several ways whereby they can determine the environment to which they are exposed at different life stages. This paper discusses three of these: plasticity in dispersal, flowering timing and germination timing. It reviews pathways through which niche construction alters evolutionary and ecological trajectories by altering the selective environment to which organisms are exposed, the phenotypic expression of plastic characters, and the expression of genetic variation. It provides examples whereby niche construction creates positive or negative feedbacks between phenotypes and environments, which in turn cause novel evolutionary constraints and novel life history expression. PMID- 15760354 TI - Maternal effects provide phenotypic adaptation to local environmental conditions. AB - In outcrossing plants, seed dispersal distance is often less than pollen movement. If the scale of environmental heterogeneity within a population is greater than typical seed dispersal distances but less than pollen movement, an individual's environment will be similar to that of its mother but not necessarily its father. Under these conditions, environmental maternal effects may evolve as a source of adaptive plasticity between generations, enhancing offspring fitness in the environment that they are likely to experience. This idea is illustrated using Campanula americana, an herb that grows in understory and light-gap habitats. Estimates of seed dispersal suggest that offspring typically experience the same light environment as their mother. In a field experiment testing the effect of open vs understory maternal light environments, maternal light directly influenced offspring germination rate and season, and indirectly affected germination season by altering maternal flowering time. Results to date indicate that these maternal effects are adaptive; further experimental tests are ongoing. Evaluating maternal environmental effects in an ecological context demonstrates that they may provide phenotypic adaptation to local environmental conditions. PMID- 15760355 TI - Evolution of phenotypic plasticity: patterns of plasticity and the emergence of ecotypes. AB - Phenotypic plasticity itself evolves, as does any other quantitative trait. A very different question is whether phenotypic plasticity causes evolution or is a major evolutionary mechanism. Existing models of the evolution of phenotypic plasticity cover many of the proposals in the literature about the role of phenotypic plasticity in evolution. I will extend existing models to cover adaptation to a novel environment, the appearance of ecotypes and possible covariation between phenotypic plasticity and mean trait value of ecotypes. Genetic assimilation does not sufficiently explain details of observed patterns. Phenotypic plasticity as a major mechanism for evolution--such as, invading new niches, speciation or macroevolution--has, at present, neither empirical nor model support. PMID- 15760356 TI - Evolutionary consequences of gender plasticity in genetically dimorphic breeding systems. AB - A functional view of gender helps evolutionary biologists evaluate the mechanisms underlying breeding-system evolution. Evolutionary pathways from hermaphroditism to dioecy include the intermediate breeding systems of gynodioecy and androdioecy. These pathways start with the invasion of unisexual mutants, females or males, respectively, followed by alteration of the hermaphrodites to allocate more to the sexual function that the unisexuals lack. Eventually, hermaphrodites become unisexual and dioecy has evolved. Some species evolving along these pathways stop short of completing this second step, or even revert back from dioecy. We evaluate the hypothesis that gender plasticity is involved in these transitions to and from dioecy. Evidence from studies of subdioecious species that have evolved along the gynodioecy pathway suggests that gender plasticity occurs and stabilizes subdioecy by lowering the cost of producing seed. Factors influencing species evolving toward androdioecy, or reverting to androdioecy from dioecy, appear to be more varied and include reproductive assurance, herbivory and gender plasticity. In general, gender specialization appears to be favored in resource-poor environments regardless of which pathway is taken to dioecy. PMID- 15760358 TI - Shade tolerance plasticity in response to neutral vs green shade cues in Polygonum species of contrasting ecological breadth. AB - Here we examined species differences in perception and response to two distinct types of shade cue, reduced photosynthetically active radiation (PAR) with and without reduced red : far red ratio (R : FR), in Polygonum persicaria and Polygonum hydropiper, two closely related annuals of contrasting ecological breadth. We compared plasticity data for light-gathering traits from glasshouse experiments at equivalently reduced PAR under neutral shade (R : FR 1.03) and green shade (R : FR 0.702). Species shared the ability to distinguish between the two types of shade, as shown by the ability of each to respond differently to neutral vs green shade for one or more traits. However, the species' responses to these cues differed significantly. Polygonum persicaria expressed stronger shade tolerance responses (increased leaf allocation and leaf area ratio) to reduced PAR alone than to green shade. By contrast, P. hydropiper expressed slightly less plasticity for these traits in neutral than in green shade. The pronounced plastic response of P. persicaria to neutral shade may contribute to the range of habitats this widespread species can occupy, which includes neutral-shade environments such as urban settings. PMID- 15760357 TI - Plasticity genes and plasticity costs: a new approach using an Arabidopsis recombinant inbred population. AB - Earlier flowering is triggered by vernalization in some but not all Arabidopsis ecotypes, often reflecting allelic variation at the FRIGIDA (FRI) locus. Using a recombinant inbred (RI) population polymorphic at FRI, we examined fitness consequences of variation for plasticity. Flowering and fitness were scored for 68 RI genotypes following full and partial vernalization treatments. Within environment and mixed-model anovas estimated variance components for a genotype effect and a G x E term, respectively. Selection analyses examined whether delayed bolting increases fitness; a plasticity costs analysis asked whether increased plasticity lowers fitness. We also explored whether trait QTL had environment-specific effects, colocated in the immediate vicinity of FRI, or overlapped with fitness QTL. Selection may favor fri alleles and constitutive early flowering, especially in conditions that only partially vernalize plants. Plasticity costs, detected only after partial vernalization and only marginally significant, were nonetheless consistent with FRI-FLC function. We discuss how information about QTL with environment-specific effects, fitness QTL, and knowledge about plasticity genes can improve interpretation of selection or plasticity cost analyses. PMID- 15760359 TI - Relationships among growth, development and plastic response to environment quality in a perennial plant. AB - Phenotypic traits differ between plants in different environments and within individuals as they grow and develop. Comparing plants in different environments at a common age can obscure the developmental basis for differences in phenotype means in different environments. Here, we compared trait means and patterns of trait ontogeny for perennial (Viola septemloba) plants growing in environments that differed in quality either naturally or due to experimental manipulation. Consistent with predictions for adaptive stress resistance, plants grown in lower quality environments allocated proportionately more biomass to roots and rhizomes, and produced smaller, thicker and longer-lived leaves. The developmental trajectory of almost all traits differed between environments, and these differences contributed to observed differences in trait means. Plants were able to alter their initial developmental trajectory in response to an increase in resources after 8 wk of growth. This result contrasts with previous findings, and may reflect a difference in the way that annual and perennial species respond to stress. Our results demonstrate the complexity of interactions between the environment and the development of the phenotype that underlie putatively adaptive plastic responses to environment quality. PMID- 15760360 TI - Optimal reproductive allocation in annuals and an informational constraint on plasticity. AB - In this computational study, we examined optimal reproductive allocation schedules in annual plants whose season lengths vary in predictability. We discuss relationships among season-length predictability, the form of the optimal allocation schedule, the degree of plasticity reflected in the optimal reaction norm, and the competitive consequences of plasticity and bet-hedging. We used an evolutionary algorithm to search the allocation-schedule space for optima, given different distributions of season length. The resulting schedules maximize geometric-mean fecundity under their selecting distributions. We then examined the relative fitness of these schedules in simulated competition among reaction norms optimized for different degrees of season-length predictability. Gradedness of optimal schedules decreases with increasing season-length predictability, and reaction norms comprising highly graded schedules reflect lesser plasticity than norms comprising schedules that are less graded. In simulations, competitively successful genotypes were those that reflected plasticity appropriate to the season-length predictability. Informational constraints in the form of low season length predictability select for low plasticity and high bet-hedging in allocation. Because an environmental cue must mediate the relationship between environment and fitness, plasticity in reproductive allocation ought to be understood not as a direct response to the selective environment, but rather to cues that are correlated with relevant environmental parameters. PMID- 15760361 TI - A comparison of phenotypic plasticity in the native dandelion Taraxacum ceratophorum and its invasive congener T. officinale. AB - We compared plastic responses to variation in the light environment for sympatric populations of native and exotic dandelion species, Taraxacum ceratophorum and Taraxacum officinale. Plasticity in leaf size, inflorescence height, reproductive phenology and dispersal-related traits were measured under experimentally altered light quality (red : far-red light ratio, R : FR) and light intensity (photosynthetically active radiation, PAR). To test whether differences in means and reaction norms of dispersal-related traits between species affected colonization potential, we created seed-dispersal models based on seed-fall rate and release height. Differences in plasticity between species were not systematic, but varied in direction and magnitude among traits. Taraxacum officinale produced larger leaves that exhibited greater plasticity in size under variable light intensity than T. ceratophorum. Plasticity in scape length at flowering occurred in relation to R : FR ratio in both species, but tended to be greater in T. ceratophorum. Seed-bearing scapes of T. officinale were taller and more canalized in height across light regimes than scapes of T. ceratophorum. Seeds of T. officinale were smaller than seeds of T. ceratophorum. Models predict greater dispersal in T. officinale within open and vegetated habitats. In contrast to the idea that plasticity promotes invasiveness, results suggest that the lack of plasticity in dispersal-related traits enhances the colonization potential of T. officinale. PMID- 15760362 TI - Expression of fission yeast cdc25 driven by the wheat ADP-glucose pyrophosphorylase large subunit promoter reduces pollen viability and prevents transmission of the transgene in wheat. AB - Cell number was to be measured in wheat (Triticum aestivum) endosperm expressing Spcdc25 (a fission yeast cell-cycle regulator) controlled by a supposedly endosperm-specific promoter, AGP2 (from the large subunit of ADP glucose pyrophosphorylase). Wheat was transformed by biolistics either with AGP2::GUS or AGP2::Spcdc25. PCR and RT-PCR checked integration and expression of the transgene, respectively. In cv. Chinese Spring, AGP2::GUS was unexpectedly expressed in carpels and pollen, as well as endosperm. In cv. Cadenza, three AGP2::Spcdc25 plants, AGP2::Spcdc25.1, .2 and .3, were generated. Spcdc25 expression was detected in mature leaves of AGP2::Spcdc25.1/.3 which exhibited abnormal spikes, 50% pollen viability and low seed set per plant; both were small compared with the nonexpressing and normal AGP2::Spcdc25.2. Spcdc25 was not transmitted to the T(1) in AGP2::Spcdc25.1 or .3, which developed normally. Spcdc25 was PCR-positive in AGP2::Spcdc25.2, using primers for a central portion, but not with primers for the 5' end, of the ORF, indicating a rearrangement; Spcdc25 was not expressed in either T(0) or T(1). The AGP2 promoter is not tissue specific and Spcdc25 expression disrupted reproduction. PMID- 15760363 TI - Seasonal adaptations of the tuberous roots of Ranunculus asiaticus to desiccation and resurrection by changes in cell structure and protein content. AB - The annual developmental cycle of tuberous roots of Ranunculus asiaticus was studied with respect to structure and content of their cells, to understand how these roots are adapted to desiccation, high temperature and rehydration. Light microscopy, histochemical analysis, and protein analyses by SDS-PAGE were employed at eight stages of annual root development. During growth and maturation of the roots, cortical cells increased in size and their cell walls accumulated pectin materials in a distinct layer to the inside of the primary walls, with pits between adjoining cells. The number of starch granules and protein bodies also increased within the cells. Several discrete proteins accumulated. Following quiescence and rehydration of the roots there was a loss of starch and proteins from the cells, and cell walls decreased in thickness. The resurrection geophyte R. asiaticus possesses desiccation-tolerant annual roots. They store carbon and nitrogen reserves within their cells, and pectin within the walls to support growth of the plant following summer quiescence and rehydration. PMID- 15760364 TI - Temperature dependence of leaf-level CO2 fixation: revising biochemical coefficients through analysis of leaf three-dimensional structure. AB - CO2 fixation in a leaf is determined by biochemical and physical processes within the boundaries set by leaf structure. Traditionally determined temperature dependencies of biochemical processes include physical processes related to CO2 exchange that result in inaccurate estimates of parameter values. A realistic three-dimensional model of a birch (Betula pendula) leaf was used to distinguish between the physical and biochemical processes affecting the temperature dependence of CO2 exchange, to determine new chloroplastic temperature dependencies for V c(max) and Jmax based on experiments, and to analyse mesophyll diffusion in detail. The constraint created by dissolution of CO2 at cell surfaces substantially decreased the CO2 flux and its concentration inside chloroplasts, especially at high temperatures. Consequently, newly determined chloroplastic V c(max) and Jmax were more temperature dependent than originally. The role of carbonic anhydrase in mesophyll diffusion appeared to be minor under representative mid-day nonwater-limited conditions. Leaf structure and physical processes significantly affect the apparent temperature dependence of CO2 exchange, especially at optimal high temperatures when the photosynthetic sink is strong. The influence of three-dimensional leaf structure on the light environment inside a leaf is marked and affects the local choice between Jmax and V c(max)-limited assimilation rates. PMID- 15760365 TI - A trade-off between nitrogen uptake and use increases responsiveness to elevated CO2 in infrequently cut mixed C3 grasses. AB - The aim of this study was to evaluate whether the responsiveness of mixed C3 grass species to elevated CO2 is related more to nitrogen uptake or to N-use efficiency. Nitrogen uptake and whole-plant N-use efficiency were investigated with two binary mixtures: Lolium perenne was mixed either with Festuca arundinacea or with Holcus lanatus. The swards were grown on sand with or without CO2 doubling, and subjected to two cutting frequencies. A C20 alcohol was used as a marker to determine species proportion in the total root mass of the mixtures. The mean residence time of N was calculated from that of 15N-labelled fertilizer. Lolium perenne took up significantly more N per unit root mass than its grass competitors, but its N-use efficiency was lower. Elevated CO2 significantly reduced the N uptake of the three grass species. A trade-off between N capture and use was found, as N-use efficiency and N-uptake rate were negatively correlated. A high N-use efficiency, and conversely low N uptake appeared to favour the responsiveness to elevated CO2 of the infrequently cut grasses. PMID- 15760366 TI - Occurrence of nodulation in unexplored leguminous trees native to the West African tropical rainforest and inoculation response of native species useful in reforestation. AB - Despite the abundance and diversity of timber tree legumes in the West African rainforest, their ability to form nitrogen-fixing nodules in symbiosis with rhizobia, and their response to rhizobial inoculation, remain poorly documented. In the first part of this study the occurrence of nodulation was determined in 156 leguminous species growing in six natural forest areas in Guinea, mostly mature trees. In the second part, an in situ experiment of rhizobial inoculation was performed on eight selected tree species belonging to three genera: Albizia, Erythrophleum and Millettia. Of the 97 plant species and 14 genera that had never been examined before this study, 31 species and four genera were reported to be nodulated. After 4 months of growing in a nursery and a further 11 months after transplantation of plants to the field, we observed a highly significant (P < 0.001) and positive effect of inoculation with Bradyrhizobium sp. strains on the growth of the eight tree species tested. The importance of determining the nodulation ability of unexplored local trees and subsequently using this information for inoculation in reforestation programmes was demonstrated. PMID- 15760367 TI - Infection of Chinese cabbage by Plasmodiophora brassicae leads to a stimulation of plant growth: impacts on cell wall metabolism and hormone balance. AB - The importance of plant hormones in clubroot infection has long been recognized. The morphological changes, such as cell division and cell elongation leading to gall formation are triggered in the early stages of infection. We analysed cell expansion by localizing Xyloglucan endoTransglucosylase/Hydrolase (XTH)-action and screened the endogenous concentrations of several classes of phytohormones by mass spectrometry in the early stages of Plasmodiophora brassicae infection in Chinese cabbage (Brassica rapa spp. pekinensis). Infected plants showed a general transient growth promotion early in infection. Furthermore a clear XTH action was visible in the epidermal layer of infected roots. Complex changes in the endogenous phytohormone profile were observed. Initially infection resulted in an increased total auxin pool. The auxin increase, together with an increased XTH action, results in wall loosening and consequently cell expansion. When the first secondary plasmodia are formed, thirteen days after infection (DAI), can be considered a switch point in phytohormone metabolism. Twenty-one DAI the plasmodia might act as a plant hormone sink resulting in a reduction in the active cytokinin pool and a lower indole-3-acetic acid content in the infected plants. PMID- 15760368 TI - Contrasting below-ground views of an ectomycorrhizal fungal community. AB - Ectomycorrhizal fungal communities have been characterized in a number of ways. Here we compare colonized root-tip and mycelia views of an ectomycorrhizal fungal community. Ectomycorrhizal fungi, both as mycelia and colonized root tips, were identified in soil samples taken from a pine plantation. We determined that for some ectomycorrhizal fungal species multiple root tips from a single soil sample were not independent. Therefore in the comparison of root-tip and mycelia views, we considered species to be present or absent from each soil sample irrespective of the number of root tips colonized by the species. We observed 39 ectomycorrhizal fungal species in total, but 12 were observed exclusively as mycelia and 11 exclusively colonizing root tips. The relative frequencies of 10 species occurring as both mycelia and root tips were not independent of the method of observation. Our results suggest that ectomycorrhizal fungal species differ in their spatial distributions on root tips, and that root-tip and mycelia views of the community are different. PMID- 15760369 TI - Phylogenetic divergence in a local population of the ectomycorrhizal fungus Cenococcum geophilum. AB - Cenococcum geophilum is a widely distributed mycorrhizal species associated with diverse gymnosperm and angiosperm hosts. In previous studies, a significant amount of genetic and genotypic diversity has been detected in this species, despite the fact that C. geophilum is not thought to reproduce by meiotic or mitotic spores. We conducted a phylogenetic analysis of 103 C. geophilum isolates from a California oak woodland and seven non-California isolates using a glyceraldehyde 3-phosphate dehydrogenase gene. In addition, a subset of isolates was analyzed using sequences from ITS-rDNA, a Group I intron located in the 3' end of the SSU-rDNA and a portion of the mitochondrial SSU-rDNA. Phylogenetically distinct lineages, or cryptic species, of C. geophilum were detected at the scale of a single soil sample within our field site. As much genetic diversity was found within a soil sample as was found for isolates collected across the USA. Our results help explain the large amount of physiological, phenotypic, and genetic differences reported among isolates of C. geophilum from similar as well as diverse geographic regions. The ecological role that these sympatric cryptic species play remains to be determined. PMID- 15760370 TI - AFLP analyses demonstrate genetic divergence, hybridization, and multiple polyploidization in the evolution of Achillea (Asteraceae-Anthemideae). AB - Achillea, a temperate genus of herbaceous allogamous perennials, is a model for evolutionary radiation through hybridization and polyploidization. AFLP analyses were performed on 300 individuals of 66 populations and 27 taxa/cytotypes, mainly from the polyploid A. millefolium aggregate and its suspected hybrid links with other clades of the genus. The mosaic genetic structure of hybrids and polyploids is revealed by specific AFLP bands shared with their assumed parents. In E Asia, A. alpina-4x and A. wilsoniana-4x are allotetraploids between A. acuminata-2x (sect. Ptarmica) and A. asiatica-2x (sect. Achillea-A. millefolium agg.). A. virescens-4x is a hybrid species linking A. nobilis agg. and A. millefolium agg. in S Europe. The hybrid swarm A. clypeolata-2x yen A. collina-4x recently formed in Bulgaria shows no AFLP bands additive to its parents; by contrast, other more ancient allopolyploids exhibit genetic innovations. Relationships within A. millefolium agg. are complex. Five 2x-taxa, mostly well separated and regressive, are limited to Eurasia; seven 4x- and 6x-taxa are intimately linked by hybridization, are expansive, and through A. asiatica-2x/4x have formed the N American polyploids. All these results from AFLPs correspond well to other evidence, and indicate a long history of reticulate evolution in Achillea. PMID- 15760371 TI - Preferential elimination of repeated DNA sequences from the paternal, Nicotiana tomentosiformis genome donor of a synthetic, allotetraploid tobacco. AB - Nicotiana tabacum (tobacco, 2n = 4x = 48) is a natural allotetraploid combining two ancestral genomes closely related to modern Nicotiana sylvestris and Nicotiana tomentosiformis. Here we examine the immediate consequences of allopolyploidy on genome evolution using 20 S4-generation plants derived from a single synthetic, S0 plant made by Burk in 1973 (Th37). Using molecular and cytogenetic methods we analysed 14 middle and highly repetitive sequences that together total approximately 4% of the genome. Two repeats related to endogenous geminiviruses (GRD5) and pararetroviruses (NtoEPRV), and two classes of satellite repeats (NTRS, A1/A2) were partially or completely eliminated at variable frequency (25-60%). These sequences are all from the N. tomentosiformis parent. Genomic in situ hybridization revealed additivity in chromosome numbers in two plants (2n = 48), while a third was aneuploid for an N. tomentosiformis-origin chromosome (2n = 49). Two plants had homozygous translocations between chromosomes of the S- and T-genomes. * The data demonstrate that genetic changes in synthetic tobacco were fast, targeted to the paternal N. tomentosiformis donated genome, and some of the changes showed concordance with changes that presumably occurred during evolution of natural tobacco. PMID- 15760372 TI - Evolution under domestication: contrasting functional morphology of seedlings in domesticated cassava and its closest wild relatives. AB - Although cassava (Manihot esculenta ssp. esculenta) is asexually propagated, farmers incorporate plants from seedlings into planting stocks. These products of sex are exposed to selection, which in agricultural environments should favour rapid growth. To examine whether seedling morphology has evolved under domestication, we compared domesticated cassava, its wild progenitor (M. esculenta ssp. flabellifolia) and their sister species (M. pruinosa) under controlled conditions. Field observations complemented laboratory study. In both wild taxa, the hypocotyl did not elongate (hypogeal germination) and cotyledons remained enclosed in the testa. In domesticated cassava, the hypocotyl elongated (epigeal germination), and cotyledons emerged and became foliaceous. The difference in hypocotyl elongation was fixed, whereas cotyledon morphology varied with environmental conditions in M. pruinosa. Comparative analysis suggests that epigeal germination is primitive in Manihot, that the lineage including wild ancestors of cassava evolved hypogeal germination--which confers greater tolerance to risks in their savanna environment--and that with domestication, there was a reversion to epigeal germination and photosynthetic cotyledons, traits conferring high initial growth rates in agricultural habitats. PMID- 15760373 TI - Statistical convenience vs biological insight: consequences of data transformation for the analysis of fitness variation in heterogeneous environments. AB - In plants, more favourable environmental conditions can lead to dramatic increases in both mean fitness and variance in fitness. This results in data that violate the equality-of-variance assumption of anova, a problem that most empiricists would address by log-transforming fitness values. Using heuristic data sets and simple simulations, we show that anova on log-transformed fitness consistently fails to match the outcome of selection in a heterogeneous environment or its sensitivity to environmental frequency. Only anova based on relative fitness within environments accurately predicts the sensitivity of genotype selection to the frequency of alternative environments. Parallel analyses of variance based on absolute fitness and relative fitness can bracket the expected success of alternative genotypes under hard and soft selection, respectively. For example, for Sinapis arvensis growing in full sun and partial shade treatments, families achieving high fitness in the best environment are favoured under hard selection, whereas soft selection favours different families that achieve consistently good performance across environments. Based on these findings, we recommend that log-transformation of fitness should no longer be standard practice in ecological genetics studies. Weighted anova is a preferable method for dealing with unequal variances, and investigators should also make greater use of techniques such as quantile regression or resampling to describe and evaluate fitness variation across heterogeneous environments. PMID- 15760374 TI - Image analysis and in vivo imaging as tools for investigation of productivity dynamics in anthocyanin-producing cell cultures of Daucus carota. AB - An anthocyanin-producing suspension culture of Daucus carota (L.) cv. Flakkese was used as model system to study secondary metabolite production in cell culture at the individual cell level. An approach was set up in which growth and production of anthocyanins were investigated using a combination of biochemical analysis, image (colour) analysis and in vivo imaging. This novel approach was used to segment the culture in different subpopulations and dissect the productive process in the cell culture grown under two different conditions, known to differ mainly for oxygen supply and mixing intensity (volume of 50 ml or 20 ml in 250 ml flasks). The 20 ml batch cultures gave a higher content and yield of anthocyanins, which depended on a complex balance between events that positively or negatively affected anthocyanin production. A model is proposed in which the different ability of cells to respond to environmental stimuli and stress depends on the different amount of anthocyanins accumulated within cells. PMID- 15760376 TI - Embryo aneuploidy screening for unexplained recurrent miscarriage: a minireview. AB - PROBLEM: The aim of this study was to investigate the incidence of chromosomal abnormalities in unexplained recurrent miscarriage (RM) patients and assess the role of pre-implantation genetic diagnosis (PGD) in preventing subsequent pregnancy loss and improving pregnancy outcome. METHOD OF STUDY: Pre-implantation genetic diagnosis was performed in 241 RM cycles and in 35 cycles in patients undergoing PGD for sex-linked diseases (control group). Chromosomes 13, 16, 18, 21, 22, X and Y were analysed by fluorescence in situ hybridization. RESULTS: The implantation and pregnancy rates in RM patients were 26.4 and 36.5% versus 20.6 and 29.0% in the control group, respectively. The percentage of abnormal embryos was significantly increased in RM patients compared with controls. CONCLUSIONS: Recurrent miscarriage is associated with a higher incidence of chromosomally abnormal embryos. In vitro fertilization (IVF) plus PGD is an important step in the management of these couples. PMID- 15760377 TI - The impact of dydrogesterone supplementation on hormonal profile and progesterone induced blocking factor concentrations in women with threatened abortion. AB - PROBLEM: The therapeutic value of progestogens in threatened abortion is still under debate. In the presence of sufficient progesterone levels during pregnancy, lymphocytes synthesize a mediator [progesterone-induced blocking factor (PIBF)] that is anti-abortive in mice. The aim of this study was to evaluate the effect of dydrogesterone on pregnancy outcome of threatened aborters. METHOD OF STUDY: Twenty-seven threatened aborters were treated for 10 days with dydrogesterone (30 40 mg/day). Sixteen healthy pregnant controls received no treatment. Serum progesterone and estradiol concentrations as well as urine PIBF concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Pregnancy outcomes in dydrogesterone-treated threatened aborters did not statistically differ from those in healthy controls. Serum progesterone concentrations in control patients, but not those in threatened aborters increased as pregnancy progressed. Following dydrogesterone treatment, initially low PIBF concentrations of threatened aborters significantly increased (P = 0.001) to reach the PIBF level found in healthy controls. CONCLUSIONS: These data suggest that by inducing PIBF production, dydrogesterone might improve pregnancy success rates in threatened aborters. PMID- 15760378 TI - Homing in on the cellular immune response to HSV-2 in humans. AB - PROBLEM: Genital herpes simplex infections are generally limited to epithelia and neurons. Vaccines have had activity in herpes simplex virus (HSV)-seronegative women only. Understanding how HSV-specific T cells traffic to infected sites may assist in vaccine design. METHOD OF STUDY: Herpes simplex virus epitopes recognized by HSV-specific CD8 T cells were identified and used to make fluorescent human leukocyte antigen (HLA)-peptide tetramers. Molecules related to lymphocyte rolling adhesion were studied by flow cytometry and cell binding. HSV specific CD4 T cells identified ex vivo by cytokine accumulation or activation marker expression, or detected in vitro by 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester (CFSE) dilution, were similarly investigated. RESULTS: Herpes simplex virus-specific T cells are 10- to 100-fold more prevalent in lesional skin compared with blood and greatly enriched in lesions compared with normal skin. Diverse viral antigens are recognized by HSV-specific T cells. Functionally active E-selectin ligand, and cutaneous lymphocyte antigen (CLA), are expressed by circulating HSV-2-specific CD8 cells. CD4 cells display lower levels of CLA that are dramatically up-regulated upon re-stimulation with antigen. CONCLUSIONS: Herpes simplex virus-2-specific CD8 and CD4 T cells differ in constitutive expression of skin homing molecules. Vaccines designed to induce proper homing are postulated to have increased efficacy. PMID- 15760379 TI - Class III-IV proliferative lupus nephritis and pregnancy: a study of 42 cases. AB - PROBLEM: A growing number of women with lupus nephritis wish pregnancy. Our aim was to analyze maternal and fetal outcome in pregnancies with the most severe forms (proliferative or class III-IV) of lupus nephritis. METHOD OF STUDY: Forty two pregnancies in 35 women with class III or IV lupus nephritis confirmed by renal biopsy (group 1); 12 pregnancies in 10 patients having histologically proven lupus nephritis class II or V (group 2); and 54 pregnant women randomly selected among our cohort of pregnant lupus patients without nephropathy who were matched for age, parity and duration of lupus to patients with class III or IV lupus nephritis (group 3) were studied. RESULTS: Pregnancy outcome and mean gestational age of neonates were similar in the three groups studied with hypertension and preeclampsia, being significantly more prevalent in patients in group 1 (37.1%) than among patients in groups 2 (11.1%) and 3 (11.6%) (P < 0.05). Mean birthweight (+/-S.D.) was significantly lower in group 1 (2214 +/- 802 g) than in groups 2 (2783 +/- 721 g) and 3 (2870 +/- 835 g) (P < 0.05). CONCLUSION: Lupus nephritis class III-IV is a risk factor for hypertensive disease during pregnancy, but it does not contraindicate gestation. PMID- 15760380 TI - Experimental genital mycoplasmosis causes increased levels of mRNA for IL-6 and TNF-alpha in the placenta. AB - PROBLEM: Previous studies with animal models have shown that injection of lipopolysaccharide (LPS) results in fetal loss and increases production of proinflammatory cytokines at the maternal-fetal interface. Most intrauterine infections, however, are associated with Ureaplasma urealyticum, a microorganism that lacks a cell wall and therefore does not contain LPS. Previous work in our laboratory with an animal model for genital infection with a similar organism, Mycoplasma pulmonis, revealed that widespread infection in maternal and fetal tissues can be experimentally induced with minimal manipulation of the animal. For this project, we tested the hypothesis that administration of the organism by a hematogenous route at gestational day (gd) 14 would result in increased tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 production by the placenta. STUDY DESIGN: Timed-pregnant, Sprague-Dawley rats were anesthetized on gd 14 and 10(7) CFU of M. pulmonis strain X1048 or an equivalent volume of sterile medium was injected into the heart. Rats were necropsied on gd 18 or 21, and ex vivo production of TNF-alpha and IL-6 was evaluated from six randomly selected placentas from each litter. The remaining placentas were harvested and either snap-frozen or placed in formalin. Frozen placentas were processed for real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of TNF-alpha and IL-6 mRNA. Formalin-fixed placentas were sectioned and stained with hematoxylin and eosin for lesion analysis. RESULTS: Concentrations of TNF-alpha but not IL-6 were significantly higher in conditioned medium from placentas harvested from infected dams at gd 21. Levels of mRNA for IL-6 and TNF-alpha, however, were increased by M. pulmonis at gd 18 and 21. Analysis of gd 21 placentas by light microscopy revealed that significant histological chorioamnionitis was present in infected animals with accumulations of neutrophils in the capsular decidua. CONCLUSION: These data indicate that experimental infection with M. pulmonis causes histological chorioamnionitis, elevated mRNA levels of TNF-alpha and IL-6 in placental tissues, and the secretion of TNF-alpha by the placenta during late gestation. PMID- 15760382 TI - Evolving issues in clinical research. PMID- 15760381 TI - CD83 monocyte-derived dendritic cells are present in human decidua and progesterone induces their differentiation in vitro. AB - PROBLEM: Dendritic cells (DCs) play an important role in antigen presentation and immunoregulation. Modifications of the immune response during pregnancy require the participation of DC. The aim of this study was to follow-up the changes of DCs in human decidua and their correlations to progesterone (Pg) concentrations. METHODS OF STUDY: Blood leukocytes were isolated from human decidua and analyzed by flow cytometry for the expression of HLA-DR and CD83 markers. Blood monocytes were cultured to differentiate and mature to DCs in the presence of Pg, estradiol or testosterone and analyzed by flow cytometry. RESULTS: The percentage of mature DCs is increased in human decidua during the 1st trimester. When Pg is added to DCs cultured in vitro the percentage of cells expressing both markers increased in specific and dose-dependent manner. CONCLUSIONS: Pg increases the numbers of monocytic DCs locally and it may be speculated that these cells help the Th1/Th2 switching in pregnancy. PMID- 15760383 TI - Steroid withdrawal for the (selected) masses. PMID- 15760384 TI - Pre-operative locoregional therapy and liver transplantation for hepatocellular carcinoma: time for a randomized controlled trial. PMID- 15760385 TI - A uniform clinical trial registration policy for journals of kidney diseases, dialysis and transplantation. PMID- 15760386 TI - Proposal for guidelines for publication of randomized trials in the American Journal of Transplantation. PMID- 15760387 TI - Proposal for the American Journal of Transplantation policy for review of ethical standards of clinical research involving live human subjects. PMID- 15760388 TI - IFN-gamma decreases CTL generation by limiting IL-2 production: A feedback loop controlling effector cell production. AB - IFN-gamma is produced by cytotoxic T lymphocytes (CTL) but can also decrease CTL generation. We used IFN-gamma-R1-deficient (GRKO) and IFN-gamma-deficient (GKO) mice to study the effects of IFN-gamma in MLC on the generation of CTL activity and CTL number, IL-2 production and cell proliferation. CTL activity was increased in MLC when GRKO responders or GKO stimulators and responders were used, compared to wild-type (WT) MLC. The number of cells displaying the CTL phenotype (CD3+, CD8+, CD25+) was also increased, accompanied by increased IL-2 production and proliferation. Combinations of WT or GRKO CD4+ T cells with WT or GRKO CD8+ T cells as responders showed that IFN-gamma mostly affects CD4+ T cells to limit CTL generation. Intracellular staining indicated that IL-2 production was largely by CD4+ T cells. Moreover, addition of IL-2 to WT responders mimicked GKO CTL generation and activity, whereas neutralizing IL-2 decreased CTL activity in GRKO and WT responders. Thus IFN-gamma reduces CTL generation in alloimmune responses largely by limiting proliferation of IL-2 producing CD4+ T cells. This creates a feedback loop in which effectors produce IFN-gamma that limits IL-2 production which in turn limits CTL generation. PMID- 15760389 TI - Selectin blockade plus therapy with low-dose sirolimus and cyclosporin a prevent brain death-induced renal allograft dysfunction. AB - Both antigen-dependent and -independent factors influence long-term organ allograft function and survival. Brain death (BD), a significant antigen independent, donor-related injury upregulates a variety of inflammatory mediators in peripheral organs. One of the earliest responses to such an insult is the expression of selectins by endothelial cells of the transplanted tissues; these in turn trigger a cascade of nonspecific events, that enhance host alloresponses and which may be worsened by toxic effects of long-term immunosuppression. Using a rat model in which donor BD accentuates subsequent renal allograft injury, we have tested the effects of therapy with recombinant P-selectin glycoprotein ligand (rPSGL-Ig) alone, or in combination with sirolimus (SRL) and cyclosporin A. We found that in contrast to the effects of standard doses of SRL or cyclosporine, rPSGL-Ig decreased inflammation in the early posttransplant period such that lower doses of maintenance immunosuppression were sufficient to maintain long-term graft function. PMID- 15760390 TI - Inhibition of obliterative airway disease development in murine tracheal allografts by matrix metalloproteinase-9 deficiency. AB - This study was designed to define the roles of matrix metalloproteinase (MMP)-2 and MMP-9 in obliterative airway disease (OAD) in heterotopic murine tracheal allografts, considered a suitable animal model for chronic lung allograft rejection. BALB/c tracheal allografts were transplanted into MMP-2-deficient (-/ ) and MMP-9-/- mice. Also, wild-type recipients were treated with doxycycline, a nonspecific MMP inhibitor. After 10, 20 and 30 days, allografts were analyzed for OAD development, intragraft levels of MMP-2 and MMP-9 and the frequency and cytokine/chemokine production profile of alloreactive T cells. Allografts transplanted into wild-type mice developed OAD lesions within 30 days. These allografts revealed significant upregulation of both MMP-2 and MMP-9. Allografts transplanted into MMP-9-/- and doxycycline-treated recipients did not develop OAD. In contrast, allografts transplanted into MMP-2-/- mice developed OAD lesions with normal kinetics. Interestingly, MMP-9-/- recipients showed an enhanced T cell alloreactivity associated with an abnormal profile of cytokine/chemokine production. The enhanced T cell alloreactivity in MMP-9-/- mice was mediated by enhanced dendritic cell stimulatory capacity as well as enhanced T cell responsive capacity. These results suggest that MMP-9 plays an important role in the pathogenesis of OAD and may represent a target for the therapeutic intervention of chronic lung allograft rejection. PMID- 15760391 TI - The hemodynamic mechanisms of lung injury and systemic inflammatory response following brain death in the transplant donor. AB - Brain-dead donors are the major source of lungs for transplantation. Brain death is characterized by two hemodynamic phases. Initially, massive sympathetic discharge results in a hypertensive crisis. This is followed by neurogenic hypotension. Up-regulation of pro-inflammatory mediators occurs in all organs and lung injury develops; this can adversely affect graft function post transplantation. The mechanisms of the systemic and lung inflammation are unknown. We hypothesized that the hemodynamic changes are responsible for these inflammatory phenomena. Brain death was induced by intra-cranial balloon inflation in rats. This resulted in hypertensive crisis, followed by hypotension. There was a significant increase in blood neutrophil CD11b/CD18 expression and pro-inflammatory cytokine levels in serum and bronchoalveolar lavage, compared with control animals. Rupture of the capillary-alveolar membrane was demonstrated by electron microscopy. Elimination of the hypertensive response by alpha adrenergic antagonist pre-treatment prevented inflammatory lung injury, reduced the systemic inflammatory markers and preserved capillary-alveolar membrane integrity. Correction of the neurogenic hypotension with noradrenaline ameliorated the systemic inflammatory response and improved oxygenation. We conclude that the sympathetic discharge triggers systemic and lung inflammation, which can be further enhanced by neurogenic hypotension. Management of the brain dead donor with early anti-inflammatory treatment and vasoconstrictors is warranted. PMID- 15760393 TI - Effect of inflammation on costimulation blockade-resistant allograft rejection. AB - Previously, we reported that allogeneic skin grafts were rapidly rejected by CD28 and CD40 ligand double deficient mice mediated by CD8+ T cells. These results indicated that some elements in addition to CD28- and CD40-mediated costimulation provide stimulatory signals for the activation of donor-specific CD8+ T cells. In this report, we investigated the role of inflammation associated with transplantation on costimulation-independent priming of CD8+ T cell during graft rejection. B6 RAG1 KO mice were transplanted with BALB/c-skin and adoptively transferred with syngeneic CD8+ T cells the same day or 50 days after transplantation. When blockade of CD28- and CD40-mediated costimulation failed to prevent acute rejection of freshly transplanted skin grafts, it efficiently delayed rejection of well-healed skin grafts. These results showed that factors associated with transplantation have essential roles in inducing costimulation blockade-resistant allograft rejection. Costimulation blockade failed to prevent acute graft-infiltration of NK cells and increasing expression of intragraft IL 12 and IL-15. These factors may trigger the graft-infiltration and priming of CD8+ T cells to induce costimulation blockade-resistant allograft rejection. PMID- 15760392 TI - Impact of MHC class II incompatibility on localization of mononuclear cell infiltrates to the bronchiolar compartment of orthotopic lung allografts. AB - Chronic pathological changes in transplanted lungs are unique because they center on the airways. We examined the relative role of MHC class I and II antigens in causing bronchial pathology in orthotopic lung transplants to rats maintained on cyclosporin A (CsA). Transplants mismatched for MHC class II antigens had significantly more peri-bronchiolar infiltrates than MHC class I incompatible transplants. No significant increase in infiltrates was found in lung transplants incompatible for MHC class I plus II antigens compared to MHC class II antigens alone. Immunohistochemistry demonstrated that MHC class II antigen expression was confined to macrophages in MHC class I incompatible transplants, but was upregulated on bronchial epithelium in transplants with MHC class II incompatibilities. Vascular endothelium was notably devoid of MHC class II antigen expression in all transplants. However, both peri-bronchial and peri vascular infiltrates were frequently cuffed by alveolar macrophages and type II pneumocytes that expressed MHC class II antigens. PCR analysis demonstrated that IFN-gamma and regulated on activation, normal T cells expressed and secreted (RANTES) were upregulated in MHC class II incompatible transplants. Thus, MHC class II incompatible orthotopic lung transplants in rats maintained on CsA immunosuppression undergo a bronchiolcentric upregulation of alloantigens. PMID- 15760394 TI - Alpha Gal ligation of pig endothelial cells induces protection from complement and apoptosis independently of NF-kappa B and inflammatory changes. AB - Cytoprotection of endothelial cells (EC) is important in EC biology and pathophysiology, including graft rejection. Using porcine aortic EC and human complement as an in vitro model of xenotransplantation, we have reported that ligation of EC Gal alpha (1-3)Gal epitopes (alpha Gal) with antibodies or lectins BS-I and IB4 induces EC resistance to injury by complement. However, before the protective response is observed, alpha Gal ligation induces an early, proinflammatory response. Using a similar model, we now investigated whether the early inflammatory response, as well as NF-kappa B activation, is required for induction of cytoprotection. Despite up-regulation of EC mRNA for many inflammatory cytokines rapidly after BS-I stimulation, recombinant cytokines or conditioned media from EC incubated with BS-I failed to induce protection when used to stimulate EC. While the lectin-induced inflammatory response was markedly reduced by inhibition of NF-kappa B, the protection from complement and apoptosis was unaffected. The lectins caused up-regulation of mRNA for protective genes A20, porcine inhibitor of apoptosis protein and hemoxygenase-1, which was not modified by NF-kappa B inhibition. These findings suggest that induction of cytoprotection in porcine EC by alpha Gal ligation results from activation of pathways that are largely independent of those that elicit NF-kappaB activation and the inflammatory response. PMID- 15760395 TI - Long-term prospective study of steroid withdrawal in kidney and heart transplant recipients. AB - A large prospective study of steroid withdrawal was performed within the framework of the Collaborative Transplant Study to analyze long-term graft and patient outcome in renal and heart transplant recipients. Steroids were withdrawn no earlier than 6 months posttransplantation. A comparison of 7-year outcomes in renal transplant recipients (94% receiving cyclosporine; 97% Caucasian) showed a benefit of steroid withdrawal versus steroid continuation in retrospectively matched controls, for graft survival (81.9% +/- 1.8% vs. 75.3% +/- 1.2%, p = 0.0001), patient survival (88.8% +/- 1.5% vs. 84.3 +/- 1.0%; p = 0.0016) and death-censored graft survival (91.8% +/- 1.3% vs. 87.9%+/- 1.0%: p = 0.0091). Steroid withdrawal was associated with improved graft survival in heart recipients also (76.2% +/- 2.4% vs. 66.9% +/- 1.7%, p = 0.0008). A total of 58.6% of renal recipients and 44.3% of heart recipients never required steroids during follow up. Rates of acute rejection and renal dysfunction did not differ between steroid-free and steroid-continuation groups. Steroid withdrawal was associated with significantly improved cardiovascular risk factors compared with steroid continuation. Rates of the development of osteoporosis and cataracts did not differ in the entire patient cohort, but were strikingly lower in patients taken off steroids during the first posttransplant year. PMID- 15760396 TI - Proteomic-based identification of cleaved urinary beta2-microglobulin as a potential marker for acute tubular injury in renal allografts. AB - Our aim is to develop noninvasive tests to monitor the renal allograft posttransplant. Previously, we have reported that an unbiased proteomic-based approach can detect urine protein peaks associated with acute tubulointerstitial renal allograft rejection. Identification of these proteins peaks by mass spectrometry demonstrated that they all derive from nontryptic cleaved forms of beta2-microglobulin. In vitro experiments showed that cleavage of intact beta2 microglobulin requires a urine pH < 6 and the presence of aspartic proteases. Patients with acute tubulointerstitial rejection had lower urine pH than stable transplants and healthy individuals. In addition, they had higher amounts of aspartic proteases and intact beta2-microglobulin in urine. These factors ultimately lead to increased amounts of cleaved urinary beta2-microglobulin. Cleaved beta2-microglobulin as an indicator of acute tubular injury may become a useful tool for noninvasive monitoring of renal allografts. PMID- 15760397 TI - Retroperitoneoscopic live donor nephrectomy (RPLDN): establishment and initial experience of RPLDN at a single center. AB - We tried to establish the technique of retroperitoneoscopic live donor nephrectomy (RPLDN). Between July 2001 and March 2004, 135 renal transplant donors underwent RPLDN. Low (average: 7 mmHg) CO2 gas pressure was employed during the procedure. All procedures were performed through a three-port retroperitoneal approach without opening the peritoneal cavity. The hand-assisted technique was not used. One hundred and twenty-seven cases were of left and eight cases were of right nephrectomy. Donor nephrectomy was carried out successfully in all patients. In one donor, the procedure was changed to open donor nephrectomy because of severe adhesion around the renal vein due to previous surgery. No serious complications, such as massive bleeding or bowel injury were encountered. Return of bowel function took 0.7 days on average. Post-operative hospital stay was 4.9 days on average, and return to work was 12 days on average. Ureteral complications occurred in 2 patients and were treated with temporally retrograde ureteral stenting. Average serum creatinine levels were 1.5 mg/dL, 1.3 mg/dL and 1.3 mg/dL at 3, 7 and 14 days after transplantation, respectively. No patients required hemodialysis after transplantation due to acute tubular necrosis. RPLDN could be an option for laparoscopic live donor nephrectomy. PMID- 15760398 TI - Evaluation of T-cell receptor repertoires in patients with long-term renal allograft survival. AB - The mechanisms underlying long-term acceptance of kidney allografts in humans under minimal or no maintenance immunosuppression are poorly understood. We analyzed the T-cell receptor (TCR) repertoires in circulating T cells of patients with long-term (> or = 9 years) renal allograft survival with (LTS-IS) and without immunosuppression (LTS-NoIS). T cells of LTS patients exhibited strongly altered TCR Vss usage, including an increased frequency of oligoclonality and a decreased frequency of polyclonality. All 3 LTS-NoIS and 12 of 16 LTS-IS patients demonstrated oligoclonality in at least three or more TCR V beta families, and the frequency of oligoclonality in these patients was significantly higher as compared to patients with well-functioning grafts at 3 years (p < 0.005 both), an uncomplicated course during the first year (p < 0.0001, both), acute rejection (p < 0.0001, both), chronic allograft nephropathy at 7 (p < 0.0001, both) or 13 years (p < 0.0001, both), dialysis patients (p < 0.0001, both) or healthy controls (p < 0.0001, both). In contrast to LTS patients, all other studied patient groups exhibited a polyclonal TCR repertoire. Our data indicate that TCR alteration is a common feature of long-term allograft outcome, which might be explained by clonal deletion, exhaustion of alloreactive T cells or predominant expression of particular T-cell subpopulations, such as regulatory T cells. PMID- 15760399 TI - The broad spectrum of quality in deceased donor kidneys. AB - The quality of the deceased donor organ clearly is one of the most crucial factors in determining graft survival and function in recipients of a kidney transplant. There has been considerable effort made towards evaluating these organs culminating in an amendment to allocation policy with the introduction of the expanded criteria donor (ECD) policy. Our study, from first solitary adult deceased donor transplant recipients from 1996 to 2002 in the National Scientific Transplant Registry database, presents a donor kidney risk grade based on significant donor characteristics, donor-recipient matches and cold ischemia time, generated directly from their risk for graft loss. We investigated the impact of our donor risk grade in a naive cohort on short- and long-term graft survival, as well as in subgroups of the population. The projected half-lives for overall graft survival in recipients by donor risk grade were I (10.7 years), II (10.0 years), III (7.9 years), IV (5.7 years) and V (4.5 years). This study indicates that there is great variability in the quality of deceased donor kidneys and that the assessment of risk might be enhanced by this scoring system as compared to the simple two-tiered system of the current ECD classification. PMID- 15760400 TI - Vascular endothelial growth factor expression and cyclosporine toxicity in renal allograft rejection. AB - The aim of this study was to evaluate the influence of vascular endothelial growth factor (VEGF) on renal function and on development of interstitial fibrosis (IF) in renal allografts. Tubular and interstitial expressions of VEGF and TNF-alpha, and density of macrophages in the interstitium were examined in 92 patients with nonrejected kidneys, acute rejection (AR), chronic allograft nephropathy (CAN), borderline changes (BC) and acute cyclosporin A (CsA) toxicity. Follow-up biopsy specimens from patients with AR and BC were evaluated for development of IF. A significant difference in tubular and interstitial VEGF expressions was found between patients with AR, BC, CAN and CsA toxicity (p < 0.001). Macrophage infiltration was positively correlated with VEGF and TNF-alpha expressions (p < 0.001). VEGF expression increased with increasing expression of TNF-alpha (p < 0.001). Renal function in first 6 months after initial biopsy was better in patients with marked tubular VEGF expression (p < 0.01); however, in follow-up, development of IF and graft loss was found earlier in these patients (p < 0.01 and p < 0.05, respectively). Increased renal VEGF expression has protective properties immediately following renal allograft but allows for increased risk of early IF, and therefore poor graft outcome in the long term. PMID- 15760401 TI - Analysis of factors that influence survival with post-transplant lymphoproliferative disorder in renal transplant recipients: the Israel Penn International Transplant Tumor Registry experience. AB - Significant mortality is associated with post-transplant lymphoproliferative disorder (PTLD) in kidney transplant recipients (KTX). Univariate/multivariate risk factor survival analysis of US PTLD KTX reported to Israel Penn International Transplant Tumor Registry from November 1968 to January 2000 was performed. PTLD presented 18 (median) (range 1-310) months in 402 KTX. Death rates were greater for those diagnosed within 6 months (64%) versus beyond 6 months (54%, p = 0.04). No differences in death risk for gender, race, immunosuppression, EBV, B or T cell positivity were identified. Death risk increased for multiple versus single sites (73% vs. 53%, hazards ratio (HR) 1.4). A 1-year increase in age increased HR for death by 2%. Surgery was associated with increased survival (55% vs. 0% without surgery) (p < 0.0001). Patients with allograft involvement, treated with transplant nephrectomy alone (n = 20), had 80% survival versus 53% without allograft removal (n = 15) (p < 0.001). Overall survival was 69% for allograft involvement alone versus 36% for other organ involvement plus allograft (n = 19 alive) (p < 0.0001). Death risk was greater for multiple site PTLD and increasing age, and risks were additive. Univariate analysis identified increased death risk for those not receiving surgery, particularly allograft involvement alone. PMID- 15760402 TI - Successful liver and kidney transplantation from cadaveric donors with left-sided bacterial endocarditis. AB - Bacterial infections are frequent in cadaveric organ donors and can be transmitted to the transplantation recipient, which could have devastating consequences for the recipients if adequate preventive measures are not adopted. From the 355 consecutive brain dead cadaveric organ donors procured at our center in the last four years, 2000-2003, four of them (1.1%) had bacterial endocarditis as cause of death. The bacteria responsible for the endocarditis were Staphylococcus epidermidis, coagulase-negative Staphylococcus, Staphylococcus hominis and Streptococcus viridans, respectively. We performed five kidney and two liver transplantations on seven recipients. All donors and recipients received antibiotic treatment against the germ causing the respective endocarditis. Infection by the bacteria responsible for the endocarditis in the respective donors was not transmitted to any of the recipients. Six of the seven recipients were alive with normal-functioning grafts after between 13 and 24 months' follow-up. Transplantectomy was performed on one kidney recipient due to thrombosis of the renal vein of the graft not related to the endocarditis. Liver and kidney transplantation from donors dying from bacterial endocarditis can be performed without causing the transmission of infection to the recipient or the dysfunction of the graft. PMID- 15760403 TI - Does additional doxorubicin chemotherapy improve outcome in patients with hepatocellular carcinoma treated by liver transplantation? AB - The aim of this prospective randomized study was to determine whether additional doxorubicin chemotherapy improves outcome in patients with hepatocellular carcinoma (HCCA) treated by liver transplantation. Stratification parameters were tumor stage (UICC I-IVa), gender, age 50 years, alpha-fetoprotein 20 ng/mL, cirrhosis and HbsAg status. For pre-operative chemotherapy doxorubicin (15 mg/m2) was given biweekly, intra-operative chemotherapy was a single dose administered before surgical manipulation. Post-operative chemotherapy from day 10 was as given preoperatively for a total dosage of 300 mg/m2. Outcome parameters were overall survival (OS) and disease-free survival. Of the 75 consecutive patients who received liver transplantation for treatment of HCCA, 62 patients were enrolled. Thirty-four patients were randomized in the chemotherapy group; 28 patients were in the control group and transplanted only. OS rates at 5 years were 38% in the chemotherapy group and 40% in the control group, disease-free survival rates at 5 years 43% and 53%, respectively. Tumor stage and vascular invasion were identified as independent risk factors for recurrence of disease. Doxorubicin chemotherapy did not improve organ survival and disease-free survival in patients undergoing liver transplantation for HCCA. PMID- 15760404 TI - The impact of pre-operative loco-regional therapy on outcome after liver transplantation for hepatocellular carcinoma. AB - No prior studies have shown that pre-operative loco-regional therapy for hepatocellular carcinoma (HCC) improves survival following orthotopic liver transplantation (OLT). We performed subgroup analyses according to pathologic HCC stage among 168 patients who underwent OLT to test the hypothesis that pre operative loco-regional therapy confers a survival advantage in a subgroup at intermediate risk for HCC recurrence. Patients with pathologic T3 HCC meeting the proposed UCSF expanded criteria (single lesion not exceeding 6.5 cm or two to three lesions none > 4.5 cm with total tumor diameter within 8 cm) had a similar 5-year recurrence-free survival as patients with pathologic T2 HCC (88.5% vs. 93.8%; p = 0.56). In the subgroup with pathologic T2 or T3 HCC, the 5-year recurrence-free survival was 93.8% for the 85 patients who received pre-operative loco-regional therapy, versus 80.6% for the other 41 patients without treatment (p = 0.049). The treatment benefit, according to 5-year recurrence-free survival, appeared greater for pathologic T3 (85.9% vs. 51.4%; p = 0.05) than T2 HCC (96.4% versus 87.1%; p = 0.12). In conclusion, although the lack of a randomized controlled design precludes drawing firm conclusions, our results suggest that pre-operative loco-regional therapy may confer a survival benefit after OLT in the subgroup with pathologic T2 and T3 HCC. PMID- 15760405 TI - A comparison of surgical outcomes and quality of life surveys in right lobe vs. left lateral segment liver donors. AB - Concern remains regarding the possibly higher risk to living liver donors of the right lobe (RL), as compared with the left lateral segment (LLS). We studied outcomes and responses to quality of life (QOL) surveys in the two groups. From 1997 to 2004, we performed 49 living donor liver transplants (LDLTs): 33 RL and 16 LLS. Notable differences included a higher proportion of female and unrelated donors in the RL group. A significantly larger liver mass was resected in RL (vs. LLS) donors: 720 (vs. 310) g, p = 0.01; RL donors also had greater blood loss (398 vs. 240 mL, p = 0.04) and operative times (7.2 vs. 5.7 h, p = 0.05). However, those findings did not translate into significant differences in donor morbidity. The complication rate was 12.5% in LLS donors and 9.1% in RL donors (p = ns). Per a QOL survey at 6 months postdonation, no significant differences were noted in SF-12 scores for the two groups. Recovery times were somewhat longer for RL donors. Mean time off work was 61.0 days for RL donors and 32.4 days for LLS donors (p = 0.004). RL donation is associated with greater operative stress for donors, but not necessarily with a more complicated recovery or differences in QOL. PMID- 15760406 TI - Preferential depletion of blood myeloid dendritic cells during acute cardiac allograft rejection under controlled immunosuppression. AB - Allo-Ag presentation to Ag-specific T-lymphocytes by donor or recipient dendritic cells (DCs) induces acute rejection (AR) after solid organ transplantation. It is postulated that myeloid (mDC) and plasmacytoid (pDC) subsets circulate differentially between bone marrow, heart and lymphoid tissues after cardiac transplantation (HTx). We investigated peripheral blood DC subset distribution, maturation and lymphoid homing properties in relation to endomyocardial biopsy (EMB) rejection grade after clinical HTx. Twenty-one HTx recipients under standard immunosuppression were studied in a 9-month follow-up. mDC and pDC numbers were analyzed by flow cytometry in fresh venous whole blood samples collected during the EMB procedures and before histological diagnosis of AR. Subsets were further characterized for maturation marker CD83 and lymphoid homing chemokine receptor CCR7. Although numbers of both DC subsets remained low for the whole post-HTx period, we observed a negative association of mDCs with rejection grade. Repeated measurements analysis revealed that only mDCs decreased during AR episodes. Rejectors had lower mDC numbers after a 3-month follow-up compared to nonrejectors. Furthermore, patients during AR exhibited low proportions of mDCs positive for CD83 or CCR7. These findings suggest peripheral blood mDC depletion in association with selective lymphoid homing of this subset during AR after clinical HTx. PMID- 15760407 TI - The roles of a bioethicist on an organ transplantation service. AB - Organ transplantation centers have expanded and increased in the last 20 years as transplant recipient outcomes have improved steadily and transplantation has moved from experimentation to treatment of choice for several indications. Transplantation presents difficult ethical and legal challenges for the transplant community and society. These include declarations of death, consent to donation and allocation of a scarce societal resource, i.e. transplantable organs. Policy and practice reflect the law, societal beliefs and prevailing values. A bioethicist contributes to a transplant team by clarifying values held by various stakeholders or embodied in decisions and policies, conducting clinical consultations, developing and interpreting policy and researching the ethics of innovations for rationing and increasing available supply of organs for transplantation. The bioethicist's interdisciplinary education, preparation, experience and familiarity with ethics, law, sociology and philosophy and skills of mediation, communication and ethical analysis contribute to addressing and resolving many issues in transplantation. This paper outlines the various roles of a bioethicist on a transplantation service, using case examples to illustrate some of the ethical issues. PMID- 15760408 TI - First experience with de novo calcineurin-inhibitor-free immunosuppression following cardiac transplantation. AB - The aim of this pilot study was to investigate whether de novo calcineurin inhibitor-free immunosuppression after cardiac transplantation is efficacious and can prevent post-operative renal impairment. Eight patients were treated by combining trough level adjusted sirolimus and mycophenolate mofetil; corticosteroids were given for the first 6 post-operative months only. Survival data, acute rejection episodes and adverse events with a special emphasis on renal impairment, myelosuppression, hypercholesterolemia, hypertriglyceridemia and infections, were recorded. With a follow-up of 3-12 months, patient survival was 100% and freedom from rejection 75%. The mean creatinine levels initially decreased and remained stable thereafter. A moderate myelosuppressive effect did not necessitate dose reduction of immunosuppressants, intermittently elevated cholesterol- and triglyceride levels decreased over time. Most frequent adverse events were pericardial effusions and peripheral edema. Complete abandonment of calcineurin inhibitor therapy by de novo use of the combination sirolimus/mycophenolate mofetil resulted in low rejection rate and avoidance of renal impairment, but should not be used without further evaluation of potential complications in a lager setting. PMID- 15760409 TI - Sharing of class I MHC molecules between donor and host promotes the infiltration of allografts by mHAg-reactive CD8 T cells. AB - Indirect recognition of minor histocompatibility antigens (mHAg) and/or MHC encoded allopeptides is an important barrier to long-term allograft acceptance following solid organ transplantation. Efficient priming of CD8+ T cells can occur after allotransplantation as a consequence of cross-presentation of donor derived proteins by the graft recipient's APC. Consistent with this, draining lymph node clonal expansion of OVA-reactive OT-I CD8+ T cells following placement of OVA-transgenic skin grafts did not depend on graft expression of K(b). However, OT-I T cells did accumulate in OVA-transgenic skin grafts most efficiently only when both the donor and host expressed K(b). OT-I infiltration of (B6-OVA x BALB/c)F1 grafts in B6 recipients was not suppressed by graft expression of H-2d. Furthermore, B6 animals transplanted with both B6-OVA and BALB/c-OVA skin had more OT-I T cells infiltrating their B6-OVA MHC-matched graft. Therefore, class I MHC matching between donor and host may not always favor an avoidance of alloreactivity within the graft tissue. PMID- 15760412 TI - Trends and results for organ donation and transplantation in the United States, 2004. PMID- 15760413 TI - Transplant data: sources, collection and research considerations, 2004. AB - The process of collecting and analyzing transplant data is complex. Familiarity with how these data are collected is crucial to a thorough understanding of the information. This article focuses on available OPTN-SRTR data and the continuing evolution of data collection mechanisms; how that data collection system is improving the data quality and reducing the data collection burden; how additional ascertainment of outcomes both completes and validates existing data; and caveats that remain for researchers. This year's article focuses further on research considerations related to cohort choice, timing of data submission, and potential biases in follow-up data. Ongoing improvements in data collection timeliness and scope are covered. The impact of extra ascertainment of outcomes, particularly for post-transplant kidney graft failure from Medicare data, are also examined. A section on graft failure reporting among different sources traces the steps by which the SRTR reconciles different data sources in its analyses. It is important that those reading and conducting transplant research understand the origin, structure, and scope of the available data. All of these issues should be carefully considered when choosing cohorts and data sources for analysis. PMID- 15760414 TI - Organ donation and utilization in the United States, 2004. AB - This article discusses issues directly related to the organ donation process, including donor consent, donor medical suitability, non-recovery of organs, organs recovered but not transplanted, expanded criteria donors (ECD), and donation after cardiac death (DCD). The findings and topics covered have important implications for how to evaluate and share best practices of organ donation as implemented by organ procurement organizations (OPOs) and major donor hospitals in the same donation service areas (DSAs). In 2002 and 2003, US hospitals referred more than one million deaths or imminent deaths to the OPOs of their DSA. Referrals increased by nearly 10% from 2002 to 2003 (1,022,280 to 1,121,392). Donor consents have increased by about 5% and the number of total deceased donors has risen from 6,187 to 6,455. Since multiple organs are recovered from most donors, this increase allowed more than 500 additional wait listed candidates to receive an organ transplant than in the prior year. Non traditional donor sources have experienced a large rate of increase; in 2003 the number of ECD kidney donors increased by 8% and the number of DCD donors increased by 43%, from 189 donors in year 2002 to 271 donors in 2003. PMID- 15760415 TI - Immunosuppression: evolution in practice and trends, 1993-2003. AB - Immunosuppression trends for solid organ transplantation have undergone a perceptible shift over the past decade. This period is of interest because it was during this time that the Food and Drug Administration (FDA) expanded the variety of medications to allow for alternatives in immunosuppressive management. An organ-by-organ review of SRTR data identifies several important trends. Antibody induction continues to be used for the majority of kidney (70%), simultaneous pancreas-kidney (SPK, 79%) pancreas after kidney (PAK, 74%), and intestine recipients (74%). It is used for under half of thoracic organ recipients and remains uncommon for liver transplant recipients (20%). The type of antibody preparation utilized has shifted from muromonab-CD3 and horse ATG to rabbit ATG and monoclonal anti-IL-2 receptor antagonists. Calcineurin inhibitors continue to be used for maintenance immunosuppression for most recipients, although there has been a shift from cyclosporine to tacrolimus. A clear transition is apparent in the choice of antimetabolite from azathioprine to mycophenolate mofetil. Although corticosteroids continue to be used as maintenance immunosuppression for most recipients prior to discharge, there is evidence that efforts of steroid avoidance protocols are having an impact across all organs, as slight decreases in their use have been observed. PMID- 15760416 TI - Pediatric transplantation, 1994-2003. AB - This article uses OPTN/SRTR data to review trends in pediatric transplantation over the last decade. In 2003, children younger than 18 made up 3% of the 82,885 candidates for organ transplantation and 7% of the 25,469 organ transplant recipients. Children accounted for 14% of the 6,455 deceased organ donors. Pediatric organ transplant recipients differ from their adult counterparts in several important aspects, including the underlying etiology of organ failure, the complexity of the surgical procedures, the pharmacokinetic properties of common immunosuppressants, the immune response following transplantation, the number and degree of comorbid conditions, and the susceptibility to post transplant complications, especially infectious diseases. Specialized pediatric organ transplant programs have been developed to address these special problems. The transplant community has responded to the particular needs of children and has provided them special consideration in the allocation of deceased donor organs. As a result of these programs and protocols, children are now frequently the most successful recipients of organ transplantation; their outcomes following kidney, liver, and heart transplantation rank among the best. This article demonstrates that substantial improvement is needed in several areas: adolescent outcomes, outcomes following intestine transplants, and waiting list mortality among pediatric heart and lung candidates. PMID- 15760417 TI - Current status of kidney and pancreas transplantation in the United States, 1994 2003. AB - This article reviews the OPTN/SRTR data collected on kidney and pancreas transplantation during 2003 in the context of trends over the past decade. Overall, the transplant community continued to struggle to meet the increasing demand for kidney and pancreas transplantation. The number of new wait-listed kidney registrants under the age of 50 has remained relatively stable since 1994, but the number of new registrants aged 50 to 64 has doubled. However, there was only a 2.3% increase in the total number of kidney transplants performed in 2003. Expanded criteria donor kidneys made up 20% of all recovered kidneys and 16% of all transplants performed, compared with 15% in the prior year. In May 2003, new rules were implemented to promote equity in kidney organ allocation. These changes seem to have improved access for historically disadvantaged groups, though they have reduced the quality of HLA matching. The effects on long-term outcomes have yet to be measured. Although the majority of SPK recipients are white (82%), the percentage of simultaneous kidney-pancreas recipients who are African-American has increased from 9% in 2000 to 16% in 2003. The percentage of Hispanic/Latino recipients increased from 5% to 9% over the same period. PMID- 15760418 TI - Liver and intestine transplantation: summary analysis, 1994-2003. AB - With nearly two years of data available since the inception of the MELD and PELD allocation system, this article examines national OPTN/SRTR data to describe trends in waiting list composition, waiting list mortality, transplant rates, and patient and graft outcomes for liver transplantation. Following a 6% reduction in the size of the waiting list after MELD was implemented in 2002, the number of patients on the waiting list grew by 2% from 2002 to 2003, while the number of liver transplants increased by 6%. The overall death rate while on the liver waiting list has decreased from 225 deaths per 1,000 patient years in 1994 to 124 deaths in 2003. As with the waiting list death rates, post-transplant death rates have also decreased over the past decade. Unadjusted one-year patient survival was lower for older donor age groups (88% for donors aged 18-34, 87% for donors aged 35-49, 85% for donors aged 50-64); a similar trend was observed at three and five years following transplantation. Intestine transplantation is performed with slowly increasing frequency and success. Early graft losses and rejection rates have changed little since 1994, but rejection is easier to control and long-term survival is improving. PMID- 15760419 TI - Thoracic organ transplantation in the United States, 1994-2003. AB - Using OPTN/SRTR data, this article reviews the state of thoracic organ transplantation in 2003 and the previous decade. Time spent on the heart waiting list has increased significantly over the last decade. The percentage of patients awaiting heart transplantation for >2 years increased from 23% in 1994 to 49% by 2003. However, there has been a general decline in heart waiting list death rates over the decade. In 2003, the lung transplant waiting list reached a record high of 3,836 registrants, up slightly from 2002 and more than threefold since 1994. One-year patient survival for those receiving lungs in 2002 was 82%, a statistically significant improvement from 2001 (78%). The number of patients awaiting a heart-lung transplant, declining since 1998, reached 189 in 2003. Adjusted patient survival for heart-lung recipients is consistently worse than the corresponding rate for isolated lung recipients, primarily due to worse outcomes for heart-lung recipients with congenital heart disease. A new lung allocation system, approved in June 2004, derives from the survival benefit of transplantation with consideration of urgency based on waiting list survival, instead of being based solely on waiting time. A goal of the policy is to minimize deaths on the waiting list. PMID- 15760420 TI - Analytical approaches for transplant research, 2004. AB - This article provides detailed explanations of the methods frequently employed in outcomes analyses performed by the Scientific Registry of Transplant Recipients (SRTR). All aspects of the analytical process are discussed, including cohort selection, post-transplant follow-up analysis, outcome definition, ascertainment of events, censoring, and adjustments. The methods employed for descriptive analyses are described, such as unadjusted mortality rates and survival probabilities, and the estimation of covariant effects through regression modeling. A section on transplant waiting time focuses on the kidney and liver waiting lists, pointing out the different considerations each list requires and the larger questions that such analyses raise. Additionally, this article describes specialized modeling strategies recently designed by the SRTR and aimed at specific organ allocation issues. The article concludes with a description of simulated allocation modeling (SAM), which has been developed by the SRTR for three organ systems: liver, thoracic organs, and kidney-pancreas. SAMs are particularly useful for comparing outcomes for proposed national allocation policies. The use of SAMs has already helped in the development and implementation of a new policy for liver candidates with high MELD scores to be offered organs regionally before the organs are offered to candidates with low MELD scores locally. PMID- 15760421 TI - Quantifying organ donation rates by donation service area. AB - Previous measures of OPO performance based on population counts have been deemed inadequate, and the need for new methods has been widely accepted. This article explains recent developments in OPO performance evaluation methodology, including those developed by the SRTR. As a replacement for the previously established measure of OPO performance--donors per million population--using eligible deaths as a national metric has yielded promising results for understanding variations in donation rates among the donation service areas assigned to each OPO. A major improvement uses "notifiable deaths" as a denominator describing a standardized maximal pool of potential donors. Notifiable deaths are defined as in-hospital deaths among ages 70 years and under, excluding certain diagnosis codes related to infections, cancers, etc. A most proximal denominator for determining donation rates is "eligible deaths," which includes only those deaths meeting the criteria for organ donation upon initial assessment. Neither measure is based on the population of a geographic unit, but on restricted upper limits of deaths that could be potential donors in any one locale (e.g., hospital or OPO). The inherent strengths and weaknesses of metrics such as donors per eligible deaths, donors per notifiable deaths, and number of organs per donor are discussed in detail. PMID- 15760422 TI - Campylobacter infections of the pericardium and myocardium. AB - Members of the genus Campylobacter are notorious for their ability to cause gastroenteritis. However, increasing numbers of case reports now suggest that they may have a wider pathogenic repertoire. Pericarditis and myocarditis are increasingly being recognised as sequelae of Campylobacter infection. Although rare, these presentations are important, as misdiagnosis may result in inappropriate thrombolysis or angioplasty, with potential accompanying complications. Extraintestinal Campylobacter infections, and the resulting pathogenesis, remain an important challenge for the 21st century, particularly as immunocompromised patients are likely to become increasingly common. PMID- 15760423 TI - Quinolones in 2005: an update. AB - Quinolones are one of the largest classes of antimicrobial agents used worldwide. This review considers the quinolones that are available currently and used widely in Europe (norfoxacin, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin) within their historical perspective, while trying to position them in the context of recent and possible future advances based on an understanding of: (1) their chemical structures and how these impact on activity and toxicity; (2) resistance mechanisms (mutations in target genes, efflux pumps); (3) their pharmacodynamic properties (AUC/MIC and Cmax/MIC ratios; mutant prevention concentration and mutant selection window); and (4) epidemiological considerations (risk of emergence of resistance, clonal spread). Their main indications are examined in relation to their advantages and drawbacks. Overall, it is concluded that these important agents should be used in an educated fashion, based on a careful balance between their ease of use and efficacy vs. the risk of emerging resistance and toxicity. However, there is now substantial evidence to support use of the most potent drug at the appropriate dose whenever this is required. PMID- 15760424 TI - Development of a real-time fluorescence resonance energy transfer PCR to identify the main pathogenic Campylobacter spp. AB - A simple real-time fluorescence resonance energy transfer (FRET) PCR, targeting the gyrA gene outside the quinolone resistance-determining region, was developed to identify Campylobacter jejuni and Campylobacter coli. These species were distinguished easily, as the corresponding melting points showed a difference of 15 degrees C. A second assay using the same biprobe and PCR conditions, but different PCR primers, was also developed to identify the less frequently encountered Campylobacter fetus. These assays were applied to 807 Campylobacter isolates from clinical specimens. Compared to phenotypic identification tests, the FRET assay yielded the same results for all except three of the isolates. Analysis by standard PCR and 16S rDNA sequencing demonstrated that two of these isolates were hippurate-negative C. jejuni strains, resulting in an erroneous phenotypic identification, while the third was an isolate of C. coli that contained a gyrA gene typical of C. jejuni, resulting in misidentification by the FRET assay. The FRET assay identified more isolates than standard PCR, which failed to yield amplification products with c. 10% of isolates. It was concluded that the FRET assays were rapid, reliable, reproducible and relatively cost efficient, as they require only one biprobe and can be performed directly on boiled isolates. PMID- 15760425 TI - Campylobacter concisus: an evaluation of certain phenotypic and genotypic characteristics. AB - The clinical relevance of Campylobacter concisus in gastrointestinal disease has not been determined definitively. This study investigated the phenotypic and genotypic characteristics of 39 C. concisus isolates from Danish patients with diarrhoea, three isolates from healthy individuals and the type strain. A cytolethal distending toxin (CDT)-like effect on Vero cells was observed in 35 (90%) isolates from patients with diarrhoea, in all three isolates from healthy individuals and in the type strain. Analysis of SDS-PAGE protein profiles and PCR amplification of 23S rDNA assigned the isolates into two distinct, but discordant groups. Automated ribotyping (RiboPrinting) identified 34 distinct patterns among the 43 isolates, but cluster analysis did not separate isolates from patients with diarrhoea from isolates from healthy patients. Random amplified polymorphic DNA (RAPD) analysis with three primers identified 37 unique profiles, but requires further evaluation. The isolates obtained from healthy carriers were distinguished by cluster analysis from the isolates obtained from patients with diarrhoea. All the isolates were susceptible to 11 antimicrobial agents tested. Overall, there was considerable variability between the C. concisus isolates, but there were no clear phenotypic or genotypic differences between isolates from patients with diarrhoea and isolates from healthy carriers. Further evidence is needed to support the possible role of C. concisus as a human enteric pathogen. PMID- 15760426 TI - Evidence for human immunodeficiency virus and Cryptococcus neoformans interactions in the pro-inflammatory and anti-inflammatory responses in blood during AIDS-associated cryptococcosis. AB - Tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, -8 and-10 and soluble TNF receptor II (sTNFR II) levels were measured at baseline, and after antifungal therapy for 2 weeks and 3 months, in plasma from 75 human immunodeficiency virus (HIV)-positive and 14 HIV-negative patients with cryptococcosis, and in plasma from 14 HIV-positive controls. At baseline, TNF alpha, IL-6 and sTNFR II levels, and cryptococcal antigen titres, were increased in patients with fungaemia compared to controls (p < 0.02). The mediator levels were not influenced by the severity of the disease or subsequent death, but sTNFR II and IL-10 levels were reduced, together with virus load, in patients receiving anti-retroviral agents (p < 0.01). During antifungal therapy, sTNFR II levels decreased (p 0.003) in parallel with the virus load and with an increase in CD4 T cell numbers. PMID- 15760428 TI - Susceptibilities to clarithromycin, amoxycillin and metronidazole of Helicobacter pylori isolates from the antrum and corpus in Tokyo, Japan, 1995-2001. AB - The aim of this study was to determine the susceptibilities to clarithromycin, amoxycillin and metronidazole of Helicobacter pylori isolates from the antrum and corpus of Japanese patients examined during the period 1995-2001. There was an increase, from 6.2% in 1995 to 22.1% in 2000-2001, in the proportion of patients infected with clarithromycin-resistant H. pylori. Of patients infected with clarithromycin-resistant H. pylori, 39.1% were infected with both clarithromycin susceptible and -resistant H. pylori. Furthermore, the MIC90 of clarithromycin for H. pylori rose from < 1 mg/L in 1995-1998 to 8 mg/L in 1999. In contrast, the MIC90s of amoxycillin and metronidazole were < or = 0.125 and 4 mg/L, respectively, throughout the study period. The results showed that, while most H. pylori isolates were susceptible to amoxycillin and metronidazole, resistance to clarithromycin among H. pylori isolates increased markedly in Japan during 1995 2001. The results also indicated a need to test the susceptibility of H. pylori isolates from more than two samples obtained from two different sites in the stomach of a single patient in order to diagnose the presence of clarithromycin resistant H. pylori correctly. PMID- 15760427 TI - Evolution of fluoroquinolone resistance among Escherichia coli urinary tract isolates from a French university hospital: application of the dynamic regression model. AB - Escherichia coli urinary tract isolates were collected in 1997-2003 from Nimes University Hospital in order to investigate long-term trends in antibiotic resistance and to explore the relationship between antibiotic use and the emergence of resistance. Time-series analysis (ARIMA models) and dynamic regression models were used to investigate relationships between antibiotic use and resistance to ofloxacin and ciprofloxacin. Significant increases were seen in the frequency of ofloxacin (8.9 to 16.7%) and ciprofloxacin resistance (6.2 to 10.1%) (p < 0.001). Using multivariate dynamic regression analysis, it was found that an increased use of one defined daily dose (DDD)/1000 patient-days for ofloxacin, ciprofloxacin and norfloxacin induced average increases of 0.81%, 0.65% and 0.53% in E. coli ofloxacin resistance (p < 0.01), with average delays of 4, 4 and 6 months, respectively. An increase of 1 DDD/1000 patient-days of ciprofloxacin, ofloxacin and norfloxacin use induced increases of 0.73%, 0.82% and 0.63% in E. coli ciprofloxacin resistance (p < 0.01), with average delays of 4, 4 and 5 months, respectively. The use of nalidixic acid was not associated significantly with an increase in resistance to fluoroquinolones by multivariate analysis. PMID- 15760429 TI - In-vitro susceptibility and molecular characterisation of macrolide resistance mechanisms among Streptococcus pneumonia isolates in The Netherlands: the DUEL 2 study. AB - In total, 881 presumptive clinical isolates of Streptococcus pneumoniae collected from throughout The Netherlands were analysed to determine their mechanisms of macrolide resistance. Isolates were identified initially by participating laboratories using their own standard identification technique, followed by determination of MICs with Etests. Only 797 isolates were confirmed as pneumococci following bile-solubility tests, lytA PCR and 16S rRNA sequencing. Of these confirmed pneumococci, 59 (7.4%) isolates were macrolide-resistant. Analysis by PCR indicated that 34 (57.6%) isolates harboured only the erm(B) gene and 16 (27.1%) only the mef gene. Three (5.1%) isolates carried both erm(B) and mef, while six (10.2%) isolates were negative for both mechanisms. Of the six negative isolates, three had a mutation in the 23S rRNA gene, and three were negative for all mechanisms tested. No isolates with the erm(A) subclass erm(TR) gene were detected. Among the 19 mef-positive isolates, 14 (73.7%) carried the mef(A) gene, and only five (26.3%) carried the mef(E) gene. No linezolid cross resistance or multiresistance (resistance to more than two classes of antibiotics) was observed. PMID- 15760430 TI - Pharmacokinetic/pharmacodynamic assessment of the in-vivo efficacy of imipenem alone or in combination with amikacin for the treatment of experimental multiresistant Acinetobacter baumannii pneumonia. AB - A guinea-pig pneumonia model involving imipenem-susceptible and imipenem resistant strains of Acinetobacter baumannii was developed to assess the in-vitro and in-vivo activities of imipenem, alone or in combination with amikacin, and the pharmacokinetic and pharmacodynamic parameters. Serum levels were measured by bioassay (imipenem) or immunoassay (amikacin), followed by calculation of pharmacokinetic and pharmacodynamic parameters (Cmax, AUC, t1/2, Cmax/MIC, AUC/MIC, and Deltat/MIC). In-vivo efficacy was evaluated by comparing bacterial counts in the lungs of treatment groups with end-of-therapy controls by anova and post-hoc tests. Decreases in the Cmax (13.4%), AUC (13%), t1/2 (25%) and Deltat/MIC (11.8-32.2%) of imipenem were observed when it was administered with amikacin, compared with administration of imipenem alone. Similarly, decreases in the Cmax (34.5%), AUC (11.6%), Cmax/MIC (34.5%) and AUC/MIC (11.7%) of amikacin were observed when it was administered with imipenem. Bacterial counts in lungs were reduced by imipenem (p 0.004) with the imipenem-susceptible strain, and by amikacin (p 0.001) with the imipenem-resistant strain. The combination of imipenem plus amikacin was inferior to imipenem alone with the imipenem susceptible strain (p 0.01), despite their in-vitro synergy, and was inferior to amikacin alone with the imipenem-resistant strain (p < 0.0001). In summary, combined use of imipenem with amikacin was less efficacious than monotherapy, probably because of a drug-drug interaction that resulted in decreased pharmacokinetic and pharmacodynamic parameters for both antimicrobial agents. PMID- 15760431 TI - The sequences of seven class D beta-lactamases isolated from carbapenem-resistant Acinetobacter baumannii from four continents. AB - Carbapenem resistance associated with class D beta-lactamases is an increasing problem in Acinetobacter baumannii. Most enzymes of this class reported so far belong to two subgroups, 1 and 2; however, a novel class D carbapenemase (OXA-51) has been reported recently which shares 56% and < 63% amino-acid identity with subgroups 1 and 2, respectively, and which belongs to a third subgroup. This study describes a further seven novel subgroup 3 beta-lactamases in carbapenem resistant A. baumannii isolates from four continents. PMID- 15760432 TI - Prevalence of Acinetobacter baumannii and other Acinetobacter spp. in faecal samples from non-hospitalised individuals. AB - In total, 226 individuals from the community were investigated for faecal carriage of Acinetobacter spp. by broth enrichment culture, followed by growth on blood agar and/or Leeds Acinetobacter Medium (LAM). Acinetobacter baumannii was isolated on both LAM and blood agar from one of 100 specimens in the UK and one of 126 specimens in The Netherlands. The predominant species were Acinetobactor johnsonii and genomic sp. 11, which were cultured from 22 and five specimens, respectively. A. baumannii did not seem to be widespread in the faecal flora of individuals in the community. PMID- 15760433 TI - Macrolide-resistance mechanisms in Streptococcus pneumoniae isolates from Belgium. AB - Of 233 erythromycin-resistant pneumococcal isolates collected in Belgium in 1999 2000, 89.7% carried the erm(B) gene, 6% the mef(A) gene, and 3.5%erm(B) plus mef(A). Two isolates contained neither erm(B) nor mef(A); one contained an erm(A) subclass erm(TR) gene, while the other contained an A2058G mutation in domain V of the 23S rRNA gene. Of 209 erm(B)-positive isolates, 191 had clindamycin MICs > 16 mg/L and 18 had MICs < or = 16 mg/L. Mef(A)-positive isolates all displayed the M resistance phenotype. Telithromycin remained active against erythromycin resistant isolates, with the highest telithromycin MIC50 being found in mef(A) positive isolates. No difference in the prevalence of different resistance mechanisms was observed compared to isolates collected in 1995-1997. PMID- 15760434 TI - Increasing erythromycin resistance among group A streptococci causing tonsillitis in a tertiary care hospital in southern India. AB - Group A streptococci (GAS) (n = 1313) isolated from patients with clinical symptoms of pharyngitis or tonsillitis attending a tertiary care hospital in southern India during 1986-2002 were tested for susceptibility to penicillin and erythromycin. The overall erythromycin resistance rate was 2.7% (n = 36). During 1986-1993, erythromycin resistance was observed in only one (2%) isolate in 1987, but reappeared in 1994 (2.7%), increased to 5.8% in 1999, and reached a maximum frequency of 13.8% in 2002. All isolates were susceptible to penicillin. The data indicate the need for continued surveillance of susceptibility patterns among GAS isolates in order to monitor the development of antibiotic resistance. PMID- 15760435 TI - Prevalence of inducible clindamycin resistance in macrolide-resistant Staphylococcus spp. AB - Between January 2002 and December 2003, macrolide-resistant isolates of Staphylococcus aureus (n = 45) and coagulase-negative staphylococci (CoNS; n = 75) from a Greek hospital were examined phenotypically for inducible clindamycin resistance. The constitutive macrolide resistance phenotype predominated (60%) in S. aureus, followed by the inducible (35%) and the clindamycin-susceptible (5%) phenotypes. In CoNS, the inducible phenotype was more common than the constitutive phenotype (50% vs. 41%). There was a significant incidence of inducible clindamycin resistance, and screening of all staphylococci is necessary in order to differentiate inducibly resistant isolates from those that are truly sensitive. PMID- 15760436 TI - The need to differentiate Campylobacter fetus subspecies isolated from humans. PMID- 15760437 TI - Progress towards meeting the challenges in clinical microbiology and infectious diseases. Proceedings of a Workshop of the European Society of Clinical Microbiology and Infectious Diseases. March 17-19, 2004. Leuven, Belgium. PMID- 15760438 TI - Surveillance and control of infectious diseases at local, national and international levels. AB - New emerging and re-emerging threats, the weight of public opinion and new technology for surveillance and treatment are likely to impact on how, and if, effective surveillance can be performed in the future. If surveillance fails to address the needs of practitioners and policy-makers, it is likely that there will be loss of confidence. Current surveillance systems are reasonably effective at detecting significant events that are localised in time and space. It is more difficult to detect diffuse and progressive events with a slow increase over time or sporadic and widespread events without obvious links to time, place or person. Detection of these events relies on good data collection, comparative background data and sophisticated analytical tools. To improve surveillance systems, we need methods with the appropriate sensitivity and specificity for the outputs desired. Targeted surveillance should enable better ascertainment of those cases which must be considered and those which can be dismissed. New methods, such as mathematical modelling and geographical information systems, support conventional surveillance in moving events into the known and predictable category. It is important to integrate surveillance across local, regional and international levels and to base surveillance on local public health structures. The purpose and value of data aggregation at each level and the amount of detail needed at each level must be carefully evaluated. The key to all these improvements is developing the workforce. Surveillance needs individuals with a broad range of skills: clinical, epidemiological, anthropological, and mathematical; in particular, people who can think laterally. These individuals must be encouraged through effective training courses, good mentorship, networking and clear career structures. PMID- 15760439 TI - Models for the organisation of hospital infection control and prevention programmes. AB - Hospital infection control is an essential part of infectious disease management and must be firmly structured and professionally organised. Prevention, limitation and eradication of nosocomial infections requires specific expertise not fully provided by clinical microbiologists and/or infectious disease consultants. Therefore, dedicated infection control physicians and nurses are essential. The basic components for successful hospital infection control include: (1) personnel and supporting resources proportional to size, complexity and estimated risk of the population served; (2) trained hospital infection control practitioners; and (3) the necessary structure to implement changes in medical, nursing and logistical organisation. The identification of areas of concern, provision of written policies and education still constitute the backbone of infection control. The infection control team must propose priorities and necessary resources, objectives, development methods, implementation and follow-up. The strategic approach must be discussed and approved by the infection control committee, comprising the hospital administrator(s), medical and nursing directors, a microbiologist, a hospital pharmacist and a delegation of clinicians. Follow-up of the projects is regularly presented to the committee by the infection control team. To what extent may evolution in the organisation of hospital infection control contribute to the optimisation of allocated resources and fulfillment of these objectives? From the Belgian experience, we conclude that structural changes represent an essential incentive. The impact of changes is greater when they are directed by the national authorities, providing resources and imposing new standards. Recommendations for staffing must consider not only the number of beds but also the objectives, complexity and characteristics of the patient population. PMID- 15760440 TI - The European Union of Medical Specialties core training curriculum in infectious diseases: overview of national systems and distribution of specialists. AB - The European Union of Medical Specialities (UEMS) Section of Infectious Diseases agreed on an infectious disease training programme in 1999, which was updated in 2002. Although the provision of infection services throughout Europe is not uniform, with variation in the roles of infectious disease physicians and microbiologists, there are, nonetheless, physicians with a predominant responsibility for clinical infectious diseases (and tropical medicine) in most countries. However, infectious diseases is formally recognised as a specific discipline by most, but not yet all, European countries. There has been consensus from national representatives to the UEMS on the content of the published curriculum. There are clear areas of overlap in training between different infection disciplines, and exploration of possible areas for closer liaison and collaboration between them has been initiated. The increased movement of medical staff within Europe will place greater demands on those responsible for training, monitoring and quality assurance. The Board and Section of Infectious Diseases have established core training programmes with a generic logbook to assist those countries without a written curriculum or record book and facilitate the development of common standards of training. The duration of training varies across Europe; 4 years is the UEMS standard, or longer if the training is combined with general internal medicine. The numbers of infection specialists (infectious diseases and microbiology) per million population show considerable variation. The UEMS Sections have recognised the importance of working closely with European specialist societies involved with training. The Section for Infectious Diseases has, in partnership with the ESCMID, established a Board for the accreditation of continuing medical education/continuing professional development. PMID- 15760441 TI - The training curriculum in hospital infection control. AB - Standardised training curricula for infection control nurses (ICNs) and recognition of the specialty exist in many European countries, but infection control physician (ICP) is not a specialty recognised by the UEMS. To gather information on curricula for ICPs, members of the ESCMID Study Group on Nosocomial Infections received a questionnaire. There is discussion about which 'professions' should be included in an infection control team. Within the 12 countries included, the average full-time equivalents (FTEs) for ICPs and ICNs per 1000 beds were 1.2 and 3.4, respectively. In addition to ICNs and ICPs, an infection control team should also include a data manager, an epidemiologist, secretarial/administrative support, and possibly, surveillance technicians. Overall, the composition of an ideal infection control team was estimated to be 9.3 FTE per 1000 beds. The background of ICPs can be clinical microbiology or infectious diseases. Among the participants, it was predominantly clinical microbiology. The ideal training curriculum for the ICP should include 6 years of postgraduate training. Of these, at least 2 years should be 'clinical training' (e.g., internal medicine) to acquire experience in the management of high-risk patients. Furthermore, training with regard to infection control and hospital epidemiology should be offered as a 'common trunk' for those being trained in clinical microbiology or infectious diseases. Important issues that remain are: implementation/standardisation of training curricula for doctors, recognition of ICP as a separate specialty or sub-specialty of clinical microbiology and/or infectious diseases, validation of on-the-job training facilities in terms of the number of doctors and nurses who can give training and the category of patients/problems present, and mandatory postgraduate education/continuing medical education specific for infection control for doctors and nurses in the field. PMID- 15760442 TI - Report of working group 1: public health challenges. AB - There is a need to improve surveillance systems in order to recognise emerging threats, both in the community and in hospitals, in a timely manner. The laboratory arm of surveillance must be complemented by hospital and primary care components. We also need more training at all levels: undergraduate and postgraduate medical training, specialist training, as well as continuing professional development schemes. Core training programmes for infectious disease specialists and medical microbiologists must be established. The contribution of general practitioners, infection specialists, microbiology laboratories, and public health specialists/agencies to response systems should be defined as clearly as possible and should be either empirically based or derived from scenario-guided calamity models. The success of surveillance hinges on sufficient long-term resources and dedicated coordination. The European Centre for Disease Prevention and Control (ECDC) may assume this role. However, structural and administrative constraints of the diverse healthcare systems throughout Europe may represent an impediment to a harmonised response. Also, the funding of the ECDC cannot cover the operational demands. The ECDC may well work as a small coordinating unit; however, without a laboratory-based scientific status, it remains doubtful whether the ECDC will achieve the competence and authority needed for effective leadership in healthcare. The ESCMID, together with other national and international scientific societies, should put emphasis on the fact that infectious diseases have not received the necessary degree of attention from governments in the European region. The ESCMID should strengthen its role in harmonising and supporting the highest standards of training in the infection disciplines. PMID- 15760443 TI - Report of working group 2: healthcare needs in the organisation and management of infection. AB - Clinical microbiology should have a physical presence, but not necessarily on site diagnostic laboratory facilities, in each hospital to ensure a quality laboratory-based infection service and strong professional interaction with clinicians. The adoption of industrial practices and the introduction of new costly molecular techniques raise the possibility that non-microbiological functions of laboratory management could be left to management professionals. This remains highly controversial; the advantages must be contrasted with the potential to disrupt the traditional managerial responsibility of the microbiologist and the links between the laboratory and clinical staff. Managers and healthcare professionals must resolve this issue, perhaps with the support of the ESCMID. Views varied, according to current professional arrangements and size of the laboratory and population served, on whether there should be a common laboratory for microbiology and other pathology disciplines with joint access to new high-technology techniques, or whether microbiology must continue as a separate facility. Clinical microbiology and infection control were viewed as core services that must be present even in smaller hospitals. Larger community hospitals and teaching centres require a full complement of expertise in laboratory and clinical practice. Integration of these disciplines within a department of infection is an emerging concept. A concern was the shortfall in trained expertise because of the ageing nature of current specialists. The importance of recruiting talented new graduates was emphasised. The importance of this topic led to a recommendation that an ESCMID working party be established to investigate the current arrangements of infection services in Europe and to make recommendations for the future organisation. PMID- 15760444 TI - Report of working group 3: specialist training and continuing medical education/professional development in the infection disciplines. AB - The European Union of Medical Specialists (UEMS) core curricula for training in infectious diseases and medical microbiology are adequate with the exception of one deficiency which is the absence of training in epidemiology, public health and infection control. Infectious disease curricula should include training in HIV, tuberculosis, hepatitis and sexually transmitted diseases. There is a need for a core curriculum in infection control. Infection control should have a basis in both medical microbiology and infectious diseases, and should become a specialty dealing with healthcare hygiene in hospitals, in outpatient clinics and also in institutions for the elderly. In the UK, a specialty training in infection is offered and includes internal medicine, clinical infectious diseases and medical microbiology for a total of 9 years. The UEMS should be contacted about the creation of a single specialty of infection, allowing for various degrees of sub-specialisation in infectious diseases or medical microbiology. It is unlikely that a European board examination validating the training of specialists will become a reality soon. Meanwhile, national systems should be created, documenting the content of the training and evaluating the quality of the training institutions. A medical specialist has a constant need for further education. This is generally a national matter, with requirements varying throughout Europe. It should be possible to accumulate continuing medical education/continuing professional development merits on a European level as well as on a national one. With the expansion of the European Union, it is important that the quality and content of specialist training can be verified and training curricula be harmonised. The UEMS should assist in this, in collaboration with scientific societies such as the ESCMID. PMID- 15760445 TI - Conclusion: ESCMID declaration on meeting the challenges in clinical microbiology and infectious diseases. PMID- 15760446 TI - Bacterial recognition and signalling by the Drosophila IMD pathway. AB - Summary Insects such as Drosophila rely entirely on innate immune responses to combat microbial pathogens. In particular, infection leads to the rapid and massive activation of anti-microbial peptide gene transcription. Drosophila utilize two NF-kappaB signalling pathways to control anti-microbial peptide gene expression, the IMD and Toll pathways. This review highlights recent advances in understanding the mechanisms of bacterial recognition utilized by both these pathways, and in deciphering the mechanisms of intracellular signalling in the IMD pathway. In particular, the peptidoglycan recognition proteins play a critical role in recognizing and discriminating different types of bacterial pathogens, and then activating either the Toll or IMD pathway. Throughout the article, the similarities and differences between Drosophila and mammalian innate immune pathways are discussed. PMID- 15760447 TI - Immune recognition of fungal beta-glucans. AB - The recognition of conserved microbial structures is a key aspect of metazoan immunity, and beta-glucans are emerging as a major target for the recognition of fungal pathogens. A number of receptors for these carbohydrates have been identified, which upon recognition, trigger a variety of immune responses. In contrast to many other systems, there is little apparent conservation in these mechanisms between vertebrates and invertebrates. In this review, we will highlight all the known receptors for beta-glucans and will discuss the various immune responses they can initiate, with reference to fungal infection, in both vertebrates and invertebrates. PMID- 15760448 TI - How C-type lectins detect pathogens. AB - Glycosylation of proteins has proven extremely important in a variety of cellular processes, including enzyme trafficking, tissue homing and immune functions. In the past decade, increasing interest in carbohydrate-mediated mechanisms has led to the identification of novel carbohydrate-recognizing receptors expressed on cells of the immune system. These non-enzymatic lectins contain one or more carbohydrate recognition domains (CRDs) that determine their specificity. In addition to their cell adhesion functions, lectins now also appear to play a major role in pathogen recognition. Depending on their structure and mode of action, lectins are subdivided in several groups. In this review, we focus on the calcium (Ca(2+))-dependent lectin group, known as C-type lectins, with the dendritic cell-specific ICAM-3 grabbing non-integrin (DC-SIGN) as a prototype type II C-type lectin organized in microdomains, and their role as pathogen recognition receptors in sensing microbes. Moreover, the cross-talk of C-type lectins with other receptors, such as Toll-like receptors, will be discussed, highlighting the emerging model that microbial recognition is based on a complex network of interacting receptors. PMID- 15760449 TI - Suppression of NF-kappaB-mediated beta-defensin gene expression in the mammalian airway by the Bordetella type III secretion system. AB - Expression of innate immune genes such as beta-defensins is induced in airway epithelium by bacterial components via activation of NF-kappaB. We show here that live Gram-negative bacteria can similarly stimulate this pathway, resulting in upregulation of the beta-defensin tracheal antimicrobial peptide (TAP) in primary cultures of bovine tracheal epithelial cells (TECs), by a Toll-like receptor 4 (TLR4)-mediated pathway. The Gram-negative airway pathogen Bordetella bronchiseptica possesses a type III secretion system previously suggested to inhibit the nuclear translocation of NF-kappaB in a cell line by immunohistochemistry. We therefore hypothesized that this pathogen might interfere in the innate immune response of the epithelium. Exposure of TECs to wild-type B. bronchiseptica suppressed the activation of NF-kappaB and the subsequent induction of TAP mRNA levels, whereas a type III secretion-defective strain did not. These results suggest a mechanism for bacterial evasion of the innate immune response in the airway, which could allow for the observed persistent colonization of this pathogen. PMID- 15760451 TI - Activation of transforming growth factor beta by Trypanosoma cruzi. AB - The anti-inflammatory cytokine, transforming growth factor beta (TGFbeta), plays an important role in Chagas disease, which is caused by the protozoan parasite Trypanosoma cruzi. In the current study, we show that the addition of an anti TGFbeta antibody inhibited T. cruzi infection of cardiomyocytes, demonstrating the requirement for active endogenous TGFbeta. As TGFbeta is synthesized as a biologically inactive precursor, which is proteolytically processed to yield a mature, active homodimer, we hypothesized that T. cruzi could activate latent TGFbeta. To test this, we added recombinant latent TGFbeta to a TGFbeta responsive reporter cell line in the presence of T. cruzi. We observed that T. cruzi was able to activate latent recombinant TGFbeta in this cellular model. We then investigated the ability of T. cruzi to activate latent TGFbetain vitro. We found that live T. cruzi, or cytosolic extracts of T. cruzi, activated latent TGFbeta in a dose- and temperature-dependent manner. The agent involved in TGFbeta activation was shown to be thermolabile and hydrophobic. Taken together, our studies demonstrate that T. cruzi directly activates latent TGFbeta. This activation is required for parasite entry into the mammalian cells and is likely to play an important role in modulating the outcome of T. cruzi infection. PMID- 15760450 TI - Role of the fungal Ras-protein kinase A pathway in governing epithelial cell interactions during oropharyngeal candidiasis. AB - Tpk1p, Tpk2p and Efg1p are members of the Ras-protein kinase A pathway that governs the yeast-to-hyphal transition in Candida albicans. We used tpk1Delta/tpk1Delta, tpk2Delta/tpk2Delta and efg1Delta/efg1Delta mutants to investigate the role of these proteins in regulating the interactions of C. albicans with oral epithelial cell lines in vitro and virulence in murine models of oropharyngeal candidiasis (OPC) and haematogenously disseminated candidiasis (HDC). The tpk1Delta/tpk1Delta strain adhered to, invaded and damaged oral epithelial cells in vitro similarly to the wild-type strain. In contrast, both the tpk2Delta/tpk2Delta and efg1Delta/efg1Delta strains had reduced capacity to invade and damage oral epithelial cells, and the efg1Delta/efg1Delta strain also exhibited decreased adherence to these cells. Consistent with these in vitro findings, the tpk2Delta/tpk2Delta and efg1Delta/efg1Delta strains also had significantly attenuated virulence during OPC. Therefore, Tpk2p and Efg1p both govern factors that enable C. albicans to invade and damage oral epithelial cells in vitro and cause OPC. These results also suggest that hyphal formation mediated by the Ras-protein kinase A pathway is a key virulence mechanism during OPC. Interestingly, the efg1Delta/efg1Delta strain, but not the tpk2Delta/tpk2Delta had reduced virulence during HDC. Thus, Tpk2p may be more important for governing virulence during OPC than HDC. PMID- 15760452 TI - Lipopolysaccharides from Helicobacter pylori can act as antagonists for Toll-like receptor 4. AB - Infection with Helicobacter pylori, a Gram-negative bacterium, is strongly associated with gastric ulcers and adenocarcinoma. The mechanisms by which the innate immune system recognizes H. pylori lipopolysaccharide (LPS) remain unclear. Contradictory reports exist that suggest that Toll-like receptors are involved. In this study we evaluated the interactions of Toll-like receptors with LPS from different strains of H. pylori. Using reporter cell lines, as well as HEK293 cells transfected with either CD14 and TLR4, or CD14 and TLR2, we show that H. pylori LPS-induced cell activation is mediated through TLR2. In addition, for the first time, we report that LPS from some H. pylori strains are able to antagonize TLR4. The antagonistic activity of H. pylori LPS from certain strains, as well as the activation via TLR2, might give H. pylori an advantage over the host that may be associated with the clinical outcome of H. pylori infection. PMID- 15760453 TI - Antiprotease inactivation by Salmonella enterica released from infected macrophages. AB - The mammalian serine protease plasmin, which has an important role in extracellular matrix degradation during cell migration, is regulated by the plasma antiprotease alpha(2)-antiplasmin (alpha(2)AP). The surface protease PgtE of Salmonella enterica serovar Typhimurium proteolytically inactivated alpha(2)AP. PgtE also activates the plasma zymogen plasminogen to plasmin, and bacteria expressing PgtE promoted degradation of extracellular matrix laminin in the presence of plasminogen and alpha(2)AP. alpha(2)AP inactivation was detected with the rough derivative of S. enterica 14028, but not with the smooth wild-type strain, suggesting that the O-antigen of lipopolysaccharide prevented contact of PgtE with the substrate molecule. After growth of S. enterica 14028 in murine J774A.1 macrophage-like cells, the infected cell lysate as well as bacteria from isolated Salmonella-containing vacuoles (SCVs) cleaved alpha(2)AP. Bacteria from SCVs produced an elevated level of PgtE and had a reduced O-antigen chain length. The lysate from S. enterica 14028-infected macrophages promoted formation of plasmin in the presence of alpha(2)AP, whereas plasmin formation by lysates from uninfected macrophages, or from macrophages infected with the pgtE-negative derivative of 14028, was inhibited by alpha(2)AP. Salmonella disseminates in the host within macrophages, which utilize plasmin for migration through tissue barriers. The results suggest that intracellular enhancement of PgtE activity in Salmonella may promote macrophage-associated proteolysis and cellular migration by altering the balance between host plasmin and alpha(2)AP. PMID- 15760454 TI - A Mycobacterium avium PPE gene is associated with the ability of the bacterium to grow in macrophages and virulence in mice. AB - PPE and PE gene families, which encode numerous proteins of unknown function, account for 10% of Mycobacterium tuberculosis genome. Mycobacterium avium genome has similar PPE and PE gene families. Using a temperature-sensitive phage phAE94 transposon mutagenesis system, a M. avium transposon library was created in the strain MAC109. Screening of individual mutants in human U937 macrophages for the ability to replicate intracellularly, we identified several attenuated clones. One of them, the 2D6 mutant, has a transposon interrupting a PPE gene (52% homologous to Rv 1787 in M. tuberculosis) was identified. The mutant and the wild type strain had comparable ability to enter macrophages. Challenge of mice with the 2D6 mutant resulted in approximately 1 log and 2 log fewer bacteria in the spleen, at 1 and 3 weeks after infection, compared with the wild-type bacterium. The 2D6 mutant grows like the wild-type bacterium in vitro. Vacuoles containing the 2D6 mutant acidified to pH 4.8; whereas, vacuoles containing wild-type bacterium were only slightly acidic. It was also observed that, in contrast to the wild-type bacterium, the 2D6 mutant did not prevent phagosome-lysosome fusion, and it is only expressed within macrophage but not in 7H9 broth. These results revealed a role for this PPE gene in the growth of M. avium in macrophages and in virulence in mice. PMID- 15760455 TI - Gene expression profiling of human promyelocytic cells in response to infection with Anaplasma phagocytophilum. AB - Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae) causes human, equine and canine granulocytic anaplasmosis and tick-borne fever of ruminants. The rickettsia parasitizes granulocytes and bone marrow progenitor cells, and can be propagated in human promyelocytic and tick cell lines. In this study, microarrays of synthetic polynucleotides of 21,329 human genes were used to identify genes that are differentially expressed in HL-60 human promyelocytic cells in response to infection with A. phagocytophilum. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) of selected genes confirmed the results of the microarray analysis. Six genes in the A. phagocytophilum-infected cells were found to be upregulated greater than 30-fold, while expression of downregulated genes most often did not change more than sixfold. Genes that were found to be differentially regulated in infected cells were those essential for cellular mechanisms including growth and differentiation, cell transport, signalling and communication and protective response against infection, some of which are most likely necessary for infection and multiplication of A. phagocytophilum in host cells. The differentially regulated genes described herein provide new information on the gene expression profiles in A. phagocytophilum-infected HL-60 cells, thus expanding in a global manner the existing information on the response of mammalian cells to A. phagocytophilum infection. PMID- 15760456 TI - Transepithelial migration of Toxoplasma gondii involves an interaction of intercellular adhesion molecule 1 (ICAM-1) with the parasite adhesin MIC2. AB - Toxoplasma gondii crosses non-permissive biological barriers such as the intestine, the blood-brain barrier and the placenta thereby gaining access to tissues where it most commonly causes severe pathology. Herein we show that in the process of migration Toxoplasma initially concentrates around intercellular junctions and probably uses a paracellular pathway to transmigrate across biological barriers. Parasite transmigration required viable and actively motile parasites. Interestingly, the integrity of host cell barriers was not altered during parasite transmigration. As intercellular adhesion molecule 1 (ICAM-1) is upregulated on cellular barriers during Toxoplasma infection, we investigated the role of this receptor in parasite transmigration. Soluble human ICAM-1 and ICAM-1 antibodies inhibited transmigration of parasites across cellular barriers implicating this receptor in the process of transmigration. Furthermore, human ICAM-1 immunoprecipitated the mature form of the parasite adhesin MIC2 present on the parasite surface, indicating that this interaction may contribute to cellular migration. These findings reveal that Toxoplasma exploits the natural cell trafficking pathways in the host to cross cellular barriers and disseminate to deep tissues. PMID- 15760457 TI - A lysine- and glutamic acid-rich protein, KERP1, from Entamoeba histolytica binds to human enterocytes. AB - Contact-dependent cytolysis of host cells by Entamoeba histolytica is an important hallmark of amoebiasis that points out the importance of molecules involved in the interaction between the parasite and the human cells. To decipher the molecular and cellular mechanisms supporting the invasion of the intestinal epithelium by E. histolytica, we analysed proteins involved in the interaction of the parasite with enterocytes. Affinity chromatography revealed several amoebic proteins interacting with purified brush border of differentiated Caco2 cells. Among them were found the intermediate subunit of the Gal/GalNAc lectin, an alpha actinin-like protein and two new proteins KERP1 and KERP2 rich in lysine and glutamic acid. In silico analysis revealed the presence of KERP2 in the closely related non-pathogenic amoeba species Entamoeba dispar but not of KERP1. In additon, polymerase chain reaction analysis allowed to suggest the absence of kerp1 homologous gene in E. dispar. Therefore, we concentrated on the cellular analysis of KERP1. Cloning of the KERP1-encoding gene, production of a recombinant protein in Escherichia coli and production of a specific antibody allowed us to show the following properties: (i) purified KERP1 binds to epithelial cell surface, (ii) KERP1 is located on the plasma membrane and in vesicles of trophozoites and (iii) KERP1 is delivered in the interstitial area between the trophozoites and the intestinal cells. PMID- 15760458 TI - Intracellular trafficking of Parachlamydia acanthamoebae. AB - Parachlamydia acanthamoebae is an obligate intracellular bacterium that naturally infects free-living amoebae. It is a potential agent of pneumonia that resists destruction by human macrophages. However, the strategy used by this Chlamydia like organism in order to resist to macrophage destruction is unknown. We analysed the intracellular trafficking of P. acanthamoebae within monocyte derived macrophages. Infected cells were immunolabelled for the bacteria and for various intracellular compartments by using specific antibodies. We analysed the bacteria colocalization with the different subcellular compartments by using epifluorescence and confocal microscopy. Bacterial replication took place 4-6 h post infection within acidic vacuoles. At that time, P. acanthamoebae colocalized with Lamp-1, a membrane marker of late endosomal and lysosomal compartments. A transient accumulation of EEA1 15 min post infection, and of rab7 and the mannose 6-phosphate receptor 30 min post infection confirmed that P. acanthamoebae traffic through the endocytic pathway. The acquisition of Lamp-1 was not different after infection with living and heat-inactivated bacteria. However, 24.5% and 79.5% of living and heat-inactivated P. acanthamoebae, respectively, colocalized with the vacuolar proton ATPase. Moreover, P. acanthamoebae did not colocalized with cathepsin D, a lysosomal hydrolase, suggesting that P. acanthamoebae interferes with maturation of its vacuole. Thus, P. acanthamoebae survives to destruction by human macrophages probably by controlling the vacuole biogenesis. PMID- 15760459 TI - Mycobacterium bovis bacille Calmette Guerin infection of human neutrophils induces CXCL8 secretion by MyD88-dependent TLR2 and TLR4 activation. AB - To investigate the potential role of neutrophils in initiation of immune responses to mycobacteria, we have characterized the response of human neutrophils to infection with Mycobacterium bovis bacille Calmette Guerin, the BCG vaccine. BCG induced transcription and secretion of the chemokine CXCL8, by signalling through Toll-like receptors TLR2 and TLR4, in conjunction with the adaptor protein myeloid differentiation factor 88 (MyD88). Blocking of responses with antibodies revealed a difference in the kinetics of signalling through the different TLRs. Anti-TLR2 antibody blocked the early phase of CXCL8 and MyD88 induction. Anti-TLR4 antibody blocked the late phase of induction occurring 2 h after infection. The existence of a TLR/MyD88 pathway for recognition and response to mycobacterial ligands provides neutrophils with the ability to drive the recruitment and activation of inflammatory cells during the early phase of mycobacterial infection and immunization. PMID- 15760462 TI - Abstracts of the 39th Annual Scientific Meeting of the European Society for Clinical Investigation, Athens, Greece, 6-9 April 2005. PMID- 15760460 TI - HGF/MET signalling protects Plasmodium-infected host cells from apoptosis. AB - Plasmodium, the causative agent of malaria, migrates through several hepatocytes before initiating a malaria infection. We have previously shown that this process induces the secretion of hepatocyte growth factor (HGF) by traversed cells, which renders neighbour hepatocytes susceptible to infection. The signalling initiated by HGF through its receptor MET has multifunctional effects on various cell types. Our results reveal a major role for apoptosis protection of host cells by HGF/MET signalling on the host susceptibility to infection. Inhibition of HGF/MET signalling induces a specific increase in apoptosis of infected cells leading to a great reduction on infection. Since HGF/MET signalling is capable of protecting cells from apoptosis by using both PI3-kinase/Akt and, to a lesser extent, MAPK pathways, we determined the impact of these pathways on Plasmodium sporozoite infection. Although inhibition of either of these pathways leads to a reduction in infection, inhibition of PI3-kinase/Akt pathway caused a stronger effect, which correlated with a higher level of apoptosis in infected host cells. Altogether, the results show that the HGF/MET signalling requirement for infection is mediated by its anti-apoptotic signal effects. These results demonstrate for the first time that active inhibition of apoptosis in host cell during infection by Plasmodium is required for a successful infection. PMID- 15760464 TI - Marine genomics: a clearing-house for genomic and transcriptomic data of marine organisms. AB - BACKGROUND: The Marine Genomics project is a functional genomics initiative developed to provide a pipeline for the curation of Expressed Sequence Tags (ESTs) and gene expression microarray data for marine organisms. It provides a unique clearing-house for marine specific EST and microarray data and is currently available at http://www.marinegenomics.org. DESCRIPTION: The Marine Genomics pipeline automates the processing, maintenance, storage and analysis of EST and microarray data for an increasing number of marine species. It currently contains 19 species databases (over 46,000 EST sequences) that are maintained by registered users from local and remote locations in Europe and South America in addition to the USA. A collection of analysis tools are implemented. These include a pipeline upload tool for EST FASTA file, sequence trace file and microarray data, an annotative text search, automated sequence trimming, sequence quality control (QA/QC) editing, sequence BLAST capabilities and a tool for interactive submission to GenBank. Another feature of this resource is the integration with a scientific computing analysis environment implemented by MATLAB. CONCLUSION: The conglomeration of multiple marine organisms with integrated analysis tools enables users to focus on the comprehensive descriptions of transcriptomic responses to typical marine stresses. This cross species data comparison and integration enables users to contain their research within a marine-oriented data management and analysis environment. PMID- 15760463 TI - Delta activity independent of its activity as a ligand of Notch. AB - BACKGROUND: Delta, Notch, and Scabrous often function together to make different cell types and refine tissue patterns during Drosophila development. Delta is known as the ligand that triggers Notch receptor activity. Scabrous is known to bind Notch and promote Notch activity in response to Delta. It is not known if Scabrous binds Delta or Delta has activity other than its activity as a ligand of Notch. It is very difficult to clearly determine this binding or activity in vivo as all Notch, Delta, and Scabrous activities are required simultaneously or successively in an inter-dependent manner. RESULTS: Using Drosophila cultured cells we show that the full length Delta promotes accumulation of Daughterless protein, fringe RNA, and pangolin RNA in the absence of Scabrous or Notch. Scabrous binds Delta and suppresses this activity even though it increases the level of the Delta intracellular domain. We also show that Scabrous can promote Notch receptor activity, in the absence of Delta. CONCLUSION: Delta has activity that is independent of its activity as a ligand of Notch. Scabrous suppresses this Delta activity. Scabrous also promotes Notch activity that is dependent on Delta's ligand activity. Thus, Notch, Delta, and Scabrous might function in complex combinatorial or mutually exclusive interactions during development. The data reported here will be of significant help in understanding these interactions in vivo. PMID- 15760465 TI - Unbiased descriptor and parameter selection confirms the potential of proteochemometric modelling. AB - BACKGROUND: Proteochemometrics is a new methodology that allows prediction of protein function directly from real interaction measurement data without the need of 3D structure information. Several reported proteochemometric models of ligand receptor interactions have already yielded significant insights into various forms of bio-molecular interactions. The proteochemometric models are multivariate regression models that predict binding affinity for a particular combination of features of the ligand and protein. Although proteochemometric models have already offered interesting results in various studies, no detailed statistical evaluation of their average predictive power has been performed. In particular, variable subset selection performed to date has always relied on using all available examples, a situation also encountered in microarray gene expression data analysis. RESULTS: A methodology for an unbiased evaluation of the predictive power of proteochemometric models was implemented and results from applying it to two of the largest proteochemometric data sets yet reported are presented. A double cross-validation loop procedure is used to estimate the expected performance of a given design method. The unbiased performance estimates (P2) obtained for the data sets that we consider confirm that properly designed single proteochemometric models have useful predictive power, but that a standard design based on cross validation may yield models with quite limited performance. The results also show that different commercial software packages employed for the design of proteochemometric models may yield very different and therefore misleading performance estimates. In addition, the differences in the models obtained in the double CV loop indicate that detailed chemical interpretation of a single proteochemometric model is uncertain when data sets are small. CONCLUSION: The double CV loop employed offer unbiased performance estimates about a given proteochemometric modelling procedure, making it possible to identify cases where the proteochemometric design does not result in useful predictive models. Chemical interpretations of single proteochemometric models are uncertain and should instead be based on all the models selected in the double CV loop employed here. PMID- 15760466 TI - Pharmacokinetic-pharmacodynamic modelling of the cardiovascular effects of drugs method development and application to magnesium in sheep. AB - BACKGROUND: There have been few reports of pharmacokinetic models that have been linked to models of the cardiovascular system. Such models could predict the cardiovascular effects of a drug under a variety of circumstances. Limiting factors may be the lack of a suitably simple cardiovascular model, the difficulty in managing extensive cardiovascular data sets, and the lack of physiologically based pharmacokinetic models that can account for blood flow changes that may be caused by a drug. An approach for addressing these limitations is proposed, and illustrated using data on the cardiovascular effects of magnesium given intravenously to sheep. The cardiovascular model was based on compartments for venous and arterial blood. Blood flowed from arterial to venous compartments via a passive flow through a systemic vascular resistance. Blood flowed from venous to arterial via a pump (the heart-lung system), the pumping rate was governed by the venous pressure (Frank-Starling mechanism). Heart rate was controlled via the difference between arterial blood pressure and a set point (Baroreceptor control). Constraints were made to pressure-volume relationships, pressure-stroke volume relationships, and physical limits were imposed to produce plausible cardiac function curves and baseline cardiovascular variables. "Cardiovascular radar plots" were developed for concisely displaying the cardiovascular status. A recirculatory kinetic model of magnesium was developed that could account for the large changes in cardiac output caused by this drug. Arterial concentrations predicted by the kinetic model were linked to the systemic vascular resistance and venous compliance terms of the cardiovascular model. The kinetic-dynamic model based on a training data set (30 mmol over 2 min) was used to predict the results for a separate validation data set (30 mmol over 5 min). RESULTS: The kinetic-dynamic model was able to describe the training data set. A recirculatory kinetic model was a good description of the acute kinetics of magnesium in sheep. The volume of distribution of magnesium in the lungs was 0.89 L, and in the body was 4.02 L. A permeability term (0.59 L min-1) described the distribution of magnesium into a deeper (probably intracellular) compartment. The final kinetic dynamic model was able to predict the validation data set. The mean prediction error for the arterial magnesium concentrations, cardiac output and mean arterial blood pressure for the validation data set were 0.02, 3.0 and 6.1%, respectively. CONCLUSION: The combination of a recirculatory model and a simple two-compartment cardiovascular model was able to describe and predict the kinetics and cardiovascular effects of magnesium in sheep. PMID- 15760467 TI - Searching QTL by gene expression: analysis of diabesity. AB - BACKGROUND: Recent developments in sequence databases provide the opportunity to relate the expression pattern of genes to their genomic position, thus creating a transcriptome map. Quantitative trait loci (QTL) are phenotypically-defined chromosomal regions that contribute to allelically variant biological traits, and by overlaying QTL on the transcriptome, the search for candidate genes becomes extremely focused. RESULTS: We used our novel data mining tool, ExQuest, to select genes within known diabesity QTL showing enriched expression in primary diabesity affected tissues. We then quantified transcripts in adipose, pancreas, and liver tissue from Tally Ho mice, a multigenic model for Type II diabetes (T2D), and from diabesity-resistant C57BL/6J controls. Analysis of the resulting quantitative PCR data using the Global Pattern Recognition analytical algorithm identified a number of genes whose expression is altered, and thus are novel candidates for diabesity QTL and/or pathways associated with diabesity. CONCLUSION: Transcription-based data mining of genes in QTL-limited intervals followed by efficient quantitative PCR methods is an effective strategy for identifying genes that may contribute to complex pathophysiological processes. PMID- 15760469 TI - Silhouette scores for assessment of SNP genotype clusters. AB - BACKGROUND: High-throughput genotyping of single nucleotide polymorphisms (SNPs) generates large amounts of data. In many SNP genotyping assays, the genotype assignment is based on scatter plots of signals corresponding to the two SNP alleles. In a robust assay the three clusters that define the genotypes are well separated and the distances between the data points within a cluster are short. "Silhouettes" is a graphical aid for interpretation and validation of data clusters that provides a measure of how well a data point was classified when it was assigned to a cluster. Thus "Silhouettes" can potentially be used as a quality measure for SNP genotyping results and for objective comparison of the performance of SNP assays at different circumstances. RESULTS: We created a program (ClusterA) for calculating "Silhouette scores", and applied it to assess the quality of SNP genotype clusters obtained by single nucleotide primer extension ("minisequencing") in the Tag-microarray format. A Silhouette score condenses the quality of the genotype assignment for each SNP assay into a single numeric value, which ranges from 1.0, when the genotype assignment is unequivocal, down to -1.0, when the genotype assignment has been arbitrary. In the present study we applied Silhouette scores to compare the performance of four DNA polymerases in our minisequencing system by analyzing 26 SNPs in both DNA polarities in 16 DNA samples. We found Silhouettes to provide a relevant measure for the quality of SNP assays at different reaction conditions, illustrated by the four DNA polymerases here. According to our result, the genotypes can be unequivocally assigned without manual inspection when the Silhouette score for a SNP assay is > 0.65. All four DNA polymerases performed satisfactorily in our Tag array minisequencing system. CONCLUSION: "Silhouette scores" for assessing the quality of SNP genotyping clusters is convenient for evaluating the quality of SNP genotype assignment, and provides an objective, numeric measure for comparing the performance of SNP assays. The program we created for calculating Silhouette scores is freely available, and can be used for quality assessment of the results from all genotyping systems, where the genotypes are assigned by cluster analysis using scatter plots. PMID- 15760468 TI - Assessment of the paraspinal muscles of subjects presenting an idiopathic scoliosis: an EMG pilot study. AB - BACKGROUND: It is known that the back muscles of scoliotic subjects present abnormalities in their fiber type composition. Some researchers have hypothesized that abnormal fiber composition can lead to paraspinal muscle dysfunction such as poor neuromuscular efficiency and muscle fatigue. EMG parameters were used to evaluate these impairments. The purpose of the present study was to examine the clinical potential of different EMG parameters such as amplitude (RMS) and median frequency (MF) of the power spectrum in order to assess the back muscles of patients presenting idiopathic scoliosis in terms of their neuromuscular efficiency and their muscular fatigue. METHODS: L5/S1 moments during isometric efforts in extension were measured in six subjects with idiopathic scoliosis and ten healthy controls. The subjects performed three 7 s ramp contractions ranging from 0 to 100% maximum voluntary contraction (MVC) and one 30 s sustained contraction at 75% MVC. Surface EMG activity was recorded bilaterally from the paraspinal muscles at L5, L3, L1 and T10. The slope of the EMG RMS/force (neuromuscular efficiency) and MF/force (muscle composition) relationships were computed during the ramp contractions while the slope of the EMG RMS/time and MF/time relationships (muscle fatigue) were computed during the sustained contraction. Comparisons were performed between the two groups and between the left and right sides for the EMG parameters. RESULTS: No significant group or side differences between the slopes of the different measures used were found at the level of the apex (around T10) of the major curve of the spine. However, a significant side difference was seen at a lower level (L3, p = 0.01) for the MF/time parameter. CONCLUSION: The EMG parameters used in this study could not discriminate between the back muscles of scoliotic subjects and those of control subject regarding fiber type composition, neuromuscular efficiency and muscle fatigue at the level of the apex. The results of this pilot study indicate that compensatory strategies are potentially seen at lower level of the spine with these EMG parameters. PMID- 15760470 TI - askMEDLINE: a free-text, natural language query tool for MEDLINE/PubMed. AB - BACKGROUND: Plain language search tools for MEDLINE/PubMed are few. We wanted to develop a search tool that would allow anyone using a free-text, natural language query and without knowing specialized vocabularies that an expert searcher might use, to find relevant citations in MEDLINE/PubMed. This tool would translate a question into an efficient search. RESULTS: The accuracy and relevance of retrieved citations were compared to references cited in BMJ POEMs and CATs (critically appraised topics) questions from the University of Michigan Department of Pediatrics. askMEDLINE correctly matched the cited references 75.8% in POEMs and 89.2 % in CATs questions on first pass. When articles that were deemed to be relevant to the clinical questions were included, the overall efficiency in retrieving journal articles was 96.8% (POEMs) and 96.3% (CATs.) CONCLUSION: askMEDLINE might be a useful search tool for clinicians, researchers, and other information seekers interested in finding current evidence in MEDLINE/PubMed. The text-only format could be convenient for users with wireless handheld devices and those with low-bandwidth connections in remote locations. PMID- 15760471 TI - Carotid intimal-media thickness as a surrogate for cardiovascular disease events in trials of HMG-CoA reductase inhibitors. AB - BACKGROUND: Surrogate measures for cardiovascular disease events have the potential to increase greatly the efficiency of clinical trials. A leading candidate for such a surrogate is the progression of intima-media thickness (IMT) of the carotid artery; much experience has been gained with this endpoint in trials of HMG-CoA reductase inhibitors (statins). METHODS AND RESULTS: We examine two separate systems of criteria that have been proposed to define surrogate endpoints, based on clinical and statistical arguments. We use published results and a formal meta-analysis to evaluate whether progression of carotid IMT meets these criteria for HMG-CoA reductase inhibitors (statins).IMT meets clinical based criteria to serve as a surrogate endpoint for cardiovascular events in statin trials, based on relative efficiency, linkage to endpoints, and congruency of effects. Results from a meta-analysis and post-trial follow-up from a single published study suggest that IMT meets established statistical criteria by accounting for intervention effects in regression models. CONCLUSION: Carotid IMT progression meets accepted definitions of a surrogate for cardiovascular disease endpoints in statin trials. This does not, however, establish that it may serve universally as a surrogate marker in trials of other agents. PMID- 15760472 TI - Daily rhythm of cerebral blood flow velocity. AB - BACKGROUND: CBFV (cerebral blood flow velocity) is lower in the morning than in the afternoon and evening. Two hypotheses have been proposed to explain the time of day changes in CBFV: 1) CBFV changes are due to sleep-associated processes or 2) time of day changes in CBFV are due to an endogenous circadian rhythm independent of sleep. The aim of this study was to examine CBFV over 30 hours of sustained wakefulness to determine whether CBFV exhibits fluctuations associated with time of day. METHODS: Eleven subjects underwent a modified constant routine protocol. CBFV from the middle cerebral artery was monitored by chronic recording of Transcranial Doppler (TCD) ultrasonography. Other variables included core body temperature (CBT), end-tidal carbon dioxide (EtCO2), blood pressure, and heart rate. Salivary dim light melatonin onset (DLMO) served as a measure of endogenous circadian phase position. RESULTS: A non-linear multiple regression, cosine fit analysis revealed that both the CBT and CBFV rhythm fit a 24 hour rhythm (R2 = 0.62 and R2 = 0.68, respectively). Circadian phase position of CBT occurred at 6:05 am while CBFV occurred at 12:02 pm, revealing a six hour, or 90 degree difference between these two rhythms (t = 4.9, df = 10, p < 0.01). Once aligned, the rhythm of CBFV closely tracked the rhythm of CBT as demonstrated by the substantial correlation between these two measures (r = 0.77, p < 0.01). CONCLUSION: In conclusion, time of day variations in CBFV have an approximately 24 hour rhythm under constant conditions, suggesting regulation by a circadian oscillator. The 90 degree-phase angle difference between the CBT and CBFV rhythms may help explain previous findings of lower CBFV values in the morning. The phase difference occurs at a time period during which cognitive performance decrements have been observed and when both cardiovascular and cerebrovascular events occur more frequently. The mechanisms underlying this phase angle difference require further exploration. PMID- 15760473 TI - The Classics of Immunology. AB - Medical Immunology will be publishing invited Reviews and Commentaries from investigators who are at the forefront of their fields, to up-date our readers as to the current state of their art. These Reviews and Commentaries will be accompanied by Editorials that place the current work into the perspective of the first contribution in an area, which resulted in a "Classic" paper. Where possible, links will be provided to the original publication, so that the modern student of immunology can read the original and draw their own conclusions as to the value of the "Classic" contribution, and its relationship to our contemporary views as to how the immune system functions. To begin this process at the very dawn of immunology, we highlight Sir Edward Jenner's first descriptions of the use of cowpox to immunize individuals against the dread disease smallpox. PMID- 15760474 TI - High-temperature unfolding of a trp-cage mini-protein: a molecular dynamics simulation study. AB - BACKGROUND: Trp cage is a recently-constructed fast-folding miniprotein. It consists of a short helix, a 3,10 helix and a C-terminal poly-proline that packs against a Trp in the alpha helix. It is known to fold within 4 ns. RESULTS: High temperature unfolding molecular dynamics simulations of the Trp cage miniprotein have been carried out in explicit water using the OPLS-AA force-field incorporated in the program GROMACS. The radius of gyration (Rg) and Root Mean Square Deviation (RMSD) have been used as order parameters to follow the unfolding process. Distributions of Rg were used to identify ensembles. CONCLUSION: Three ensembles could be identified. While the native-state ensemble shows an Rg distribution that is slightly skewed, the second ensemble, which is presumably the Transition State Ensemble (TSE), shows an excellent fit. The denatured ensemble shows large fluctuations, but a Gaussian curve could be fitted. This means that the unfolding process is two-state. Representative structures from each of these ensembles are presented here. PMID- 15760476 TI - Ethmoidal osteoid osteoma with orbital and intracranial extension - a case report. AB - BACKGROUND: Osteoid osteoma is a benign bone neoplasm which is seen in the long bones of appendicular skeleton. It is rarely seen in the cranium. Skull base osteoid osteoma is extremely rare and has been anecdotally reported. CASE PRESENTATION: The authors report a case of a large osteoid osteoma of the ethmoid with intraorbital and intracranial extension in a 33 year old male patient. He presented with loss of vision in the left eye. The intra-cranial extension was excised through a single burr-hole fronto-orbitotomy. The ethmoid and orbital portions were approached and excised through a Weber-Ferguson incision and inferior orbitotomy. Radical excision of the tumor could thus be achieved through a craniofacial approach. CONCLUSION: Although benign and rare, skull base osteoid osteoma can present with neurological deficit due to its mass effect and involvement of vital structures. A multispeciality team approach is advisable in such cases if radical excision is to be achieved. A craniofacial approach made radical single stage excision of this multicompartmental osteoid osteoma possible with an uneventful postoperative period. PMID- 15760475 TI - Analysis of phosphorylation of human heat shock factor 1 in cells experiencing a stress. AB - BACKGROUND: Heat shock factor (HSF/HSF1) not only is the transcription factor primarily responsible for the transcriptional response of cells to physical and chemical stress but also coregulates other important signaling pathways. The factor mediates the stress-induced expression of heat shock or stress proteins (HSPs). HSF/HSF1 is inactive in unstressed cells and is activated during stress. Activation is accompanied by hyperphosphorylation of the factor. The regulatory importance of this phosphorylation has remained incompletely understood. Several previous studies on human HSF1 were concerned with phosphorylation on Ser303, Ser307 and Ser363, which phosphorylation appears to be related to factor deactivation subsequent to stress, and one study reported stress-induced phosphorylation of Ser230 contributing to factor activation. However, no previous study attempted to fully describe the phosphorylation status of an HSF/HSF1 in stressed cells and to systematically identify phosphoresidues involved in factor activation. The present study reports such an analysis for human HSF1 in heat stressed cells. RESULTS: An alanine scan of all Ser, Thr and Tyr residues of human HSF1 was carried out using a validated transactivation assay, and residues phosphorylated in HSF1 were identified by mass spectrometry and sequencing. HSF1 activated by heat treatment was phosphorylated on Ser121, Ser230, Ser292, Ser303, Ser307, Ser314, Ser319, Ser326, Ser344, Ser363, Ser419, and Ser444. Phosphorylation of Ser326 but none of the other Ser residues was found to contribute significantly to activation of the factor by heat stress. Phosphorylation on Ser326 increased rapidly during heat stress as shown by experiments using a pSer326 phosphopeptide antibody. Heat stress-induced DNA binding and nuclear translocation of a S326A substitution mutant was not impaired in HSF1-negative cells, but the mutant stimulated HSP70 expression several times less well than wild type factor. CONCLUSION: Twelve Ser residues but no Thr or Tyr residues were identified that were phosphorylated in heat-activated HSF1. Mutagenesis experiments and functional studies suggested that phosphorylation of HSF1 residue Ser326 plays a critical role in the induction of the factor's transcriptional competence by heat stress. PhosphoSer326 also contributes to activation of HSF1 by chemical stress. To date, no functional role could be ascribed to any of the other newly identified phosphoSer residues. PMID- 15760477 TI - Two new plumage mutations in the Japanese quail: "curly" feather and "rusty" plumage. AB - BACKGROUND: The genetics of plumage of Japanese quail is of interest both from a biological standpoint, for comparative studies between avian species, and from a zootechnical standpoint, for identifying commercial selection lines or crosses. There are only few plumage mutations reported in quail, and the present work describes a new color variant "rusty" and a new feather structure "curly", and their heredity from an F1 and F2 segregation experiment. RESULTS: Curly feathers result from abnormal early growth caused by transient joining of follicle walls of adjacent feathers around 10 days of age, but the expression of the trait is variable. Rusty plumage color results from the replacement of the wild-type plumage pattern on the tip of the feather by a reddish coloration, but the pigmentation of the bottom part of the feather is not affected. Two lines breeding true for the curly or the rusty phenotype were developed. Both characters are determined by autosomal recessive mutations which are independent. The curly mutation has also a positive effect on body weight at 5 weeks of age. CONCLUSION: The curly line is a new model which may be used for further work on the growth of the feather, and the rusty mutation is a new addition to the panel of plumage mutations available for comparative studies in poultry, and more generally among avian species. PMID- 15760479 TI - Isolation by distance, web service. AB - BACKGROUND: The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user friendly. RESULTS: We have created Isolation by Distance Web Service (IBDWS) a user-friendly web interface for determining patterns of isolation by distance. Using this site, population geneticists can perform a variety of powerful statistical tests including Mantel tests, Reduced Major Axis (RMA) regression analysis, as well as calculate FST between all pairs of populations and perform basic summary statistics (e.g., heterozygosity). All statistical results, including publication-quality scatter plots in Postscript format, are returned rapidly to the user and can be easily downloaded. CONCLUSION: IBDWS population genetics analysis software is hosted at http://phage.sdsu.edu/~jensen/ and documentation is available at http://www.bio.sdsu.edu/pub/andy/IBD.html. The source code has been made available on Source Forge at http://sourceforge.net/projects/ibdws/. PMID- 15760478 TI - CoPub Mapper: mining MEDLINE based on search term co-publication. AB - BACKGROUND: High throughput microarray analyses result in many differentially expressed genes that are potentially responsible for the biological process of interest. In order to identify biological similarities between genes, publications from MEDLINE were identified in which pairs of gene names and combinations of gene name with specific keywords were co-mentioned. RESULTS: MEDLINE search strings for 15,621 known genes and 3,731 keywords were generated and validated. PubMed IDs were retrieved from MEDLINE and relative probability of co-occurrences of all gene-gene and gene-keyword pairs determined. To assess gene clustering according to literature co-publication, 150 genes consisting of 8 sets with known connections (same pathway, same protein complex, or same cellular localization, etc.) were run through the program. Receiver operator characteristics (ROC) analyses showed that most gene sets were clustered much better than expected by random chance. To test grouping of genes from real microarray data, 221 differentially expressed genes from a microarray experiment were analyzed with CoPub Mapper, which resulted in several relevant clusters of genes with biological process and disease keywords. In addition, all genes versus keywords were hierarchical clustered to reveal a complete grouping of published genes based on co-occurrence. CONCLUSION: The CoPub Mapper program allows for quick and versatile querying of co-published genes and keywords and can be successfully used to cluster predefined groups of genes and microarray data. PMID- 15760481 TI - Commentary: physiology is the logic of life. PMID- 15760480 TI - Ca2+ regulation in the absence of the iplA gene product in Dictyostelium discoideum. AB - BACKGROUND: Stimulation of Dictyostelium discoideum with cAMP evokes an elevation of the cytosolic free Ca2+ concentration ([Ca2+]i). The [Ca2+]i-change is composed of liberation of stored Ca2+ and extracellular Ca2+-entry. The significance of the [Ca2+]i-transient for chemotaxis is under debate. Abolition of chemotactic orientation and migration by Ca2+-buffers in the cytosol indicates that a [Ca2+]i-increase is required for chemotaxis. Yet, the iplA- mutant disrupted in a gene bearing similarity to IP3-receptors of higher eukaryotes aggregates despite the absence of a cAMP-induced [Ca2+]i-transient which favours the view that [Ca2+]i-changes are insignificant for chemotaxis. RESULTS: We investigated Ca2+-fluxes and the effect of their disturbance on chemotaxis and development of iplA- cells. Differentiation was altered as compared to wild type amoebae and sensitive towards manipulation of the level of stored Ca2+. Chemotaxis was impaired when [Ca2+]i-transients were suppressed by the presence of a Ca2+-chelator in the cytosol of the cells. Analysis of ion fluxes revealed that capacitative Ca2+-entry was fully operative in the mutant. In suspensions of intact and permeabilized cells cAMP elicited extracellular Ca2+-influx and liberation of stored Ca2+, respectively, yet to a lesser extent than in wild type. In suspensions of partially purified storage vesicles ATP-induced Ca2+ uptake and Ca2+-release activated by fatty acids or Ca2+-ATPase inhibitors were similar to wild type. Mn2+-quenching of fura2 fluorescence allows to study Ca2+ influx indirectly and revealed that the responsiveness of mutant cells was shifted to higher concentrations: roughly 100 times more Mn2+ was necessary to observe agonist-induced Mn2+-influx. cAMP evoked a [Ca2+]i-elevation when stores were strongly loaded with Ca2+, again with a similar shift in sensitivity in the mutant. In addition, basal [Ca2+]i was significantly lower in iplA- than in wild type amoebae. CONCLUSION: These results support the view that [Ca2+]i-transients are essential for chemotaxis and differentiation. Moreover, capacitative and agonist-activated ion fluxes are regulated by separate pathways that are mediated either by two types of channels in the plasma membrane or by distinct mechanisms coupling Ca2+-release from stores to Ca2+-entry in Dictyostelium. The iplA- strain retains the capacitative Ca2+-entry pathway and an impaired agonist activated pathway that operates with reduced efficiency or at higher ionic pressure. PMID- 15760482 TI - The modeling of oxidative phosphorylation in skeletal muscle. AB - A computer model of oxidative phosphorylation was developed in isolated muscle mitochondria [Korzeniewski and Mazat: Biochem J 319: 143-148, 1996] and in intact skeletal muscle [Korzeniewski and Zoladz: Biophys Chem 92: 17-34, 2001]. Within this model the dependence on different metabolite concentrations of the rate of each enzymatic reaction, process and flux is described by an appropriate kinetic equation. The changes of metabolite concentrations over time are described by a set of ordinary differential equations. The model has been very extensively tested by a comparison of computer simulations with a broad set of experimental results concerning various kinetic properties of the oxidative phosphorylation system. Next the model was used for theoretical studies on the regulation of oxidative phosphorylation in intact muscle cells. The model decidedly supports the so-called parallel-activation mechanism or each-step-activation mechanism of adjusting the rate of ATP supply to the current energy demand [Korzeniewski: Biochem J 330: 1189-1195, 1998; Korzeniewski: Biochem J 375: 799-804, 2003]. Because of this mechanism, not only ATP usage, but also the substrate dehydrogenation system and all oxidative phosphorylation complexes (complex I, complex III, complex IV, ATP synthase, ATP/ADP carrier, phosphate carrier) are directly (and not by changes in metabolite concentrations) activated by some intracellular factor(s) related to muscle contraction, probably by calcium ions, during the transition from rest to work. This mechanism is able to account for several kinetic properties of oxidative phosphorylation that cannot be explained by other mechanisms postulated in the literature. Thus the discussed kinetic model of oxidative phosphorylation has appeared to be a very useful research tool. PMID- 15760483 TI - An in silico study of energy metabolism in cardiac excitation-contraction coupling. AB - The heart produces and uses ATP at a high rate. Each step involved in ATP metabolism has been extensively studied. However, functional coupling between ATP production and membrane excitation-contraction coupling, which is the main ATP consumption process, is not yet fully understood because of complicated interactions and the lack of quantitative data obtained in vivo. Computer simulation is a powerful tool for integrating experimental data and for solving their complicated interactions. To investigate the mechanisms underlying cardiac excitation-contraction-energy metabolism coupling, we have developed a computer model of cardiac excitation-contraction coupling (Kyoto model) that includes the major processes of ATP production, such as oxidative phosphorylation that was originally developed for skeletal muscle by Korzeniewski and Zoladz [Biophys Chem 92: 17-34, 2001], creatine kinase, and adenylate kinase. In this review, we briefly summarize cardiac energy metabolism and discuss the regulation of mitochondrial ATP synthesis, using the Kyoto model. PMID- 15760484 TI - Computational modeling of cardiac ventricular action potentials in rat and mouse: review. AB - Little is known about the ionic mechanisms underlying the action potential heterogeneity in ventricle-associated healthy and disease conditions, even though five decades of histological, electrophysiological, pharmacological, and biochemical investigations exist. The computational modeling in murine ventricular myocytes can complement our knowledge of the experimental data and provide us with more quantitative descriptions in understanding different conditions related to normal and disease conditions. This paper initially reviews the theoretical modeling for cardiac ventricular action potentials of various species and the related experimental work. It then presents the progress of the computational modeling of cardiac ventricular cells for normal, diabetic, and spontaneously hypertensive rats. The paper also introduces recent modeling efforts for the action potential heterogeneity in mouse ventricular cells. The computational insights gained into the ionic mechanisms in rodents will continue to enhance our understanding of the heart and provide us with new knowledge for future studies to treat cardiac diseases in children and adults. Because the dissemination of computational models is very important, we continue to disseminate these models by iCell, the interactive cell modeling resource. iCell (http://ssd1.bme.memphis.edu/icell/) has been developed as a simulation-based teaching and learning tool for electrophysiology and contains JAVA applets that present models of various cardiac cells and neurons and simulation data of their bioelectric activities at cellular level. PMID- 15760485 TI - Functionally and structurally integrated computational modeling of ventricular physiology. AB - Computational biology is integrative in several ways. Functionally, computational models are valuable for integrating the many interacting processes within biochemical networks and the many interacting physiological subsystems within the cell. Structurally detailed models provide a way of integrating across scales of biological organization from molecule to organism. Data integration across diverse laboratory and clinical measurements is another unique strength of computational biology. We describe examples of all three categories of integration by using recent advances in modeling cardiac excitation-contraction coupling and whole-heart electromechanics in health and disease. PMID- 15760486 TI - Giving form to the function of the heart: embedding cellular models in an anatomical framework. AB - A computational framework is presented for integrating the electrical, mechanical, and biochemical functions of the heart. The construction of efficient finite element representations of canine and porcine ventricular geometry and microstructure is outlined. Computational techniques are applied to solve large deformation soft tissue mechanics by using orthotropic constitutive laws for myocardial tissue and models of active tension generation embedded at the Gauss points in the finite element mesh. The reaction-diffusion equations governing electrical current flow in the heart are solved on a grid of deforming material points that access systems of ordinary differential equations representing the cellular processes underlying the cardiac action potential. Navier-Stokes equations are solved to predict coronary blood flow in a system of branching blood vessels embedded in the deforming myocardium. PMID- 15760487 TI - Mathematical modeling of cardiovascular system dynamics using a lumped parameter method. AB - This work reviews the main aspects of cardiovascular system dynamics with an emphasis on modeling hemodynamic characteristics by the use of a lumped parameter approach. The methodological and physiological aspects of the circulation dynamics are summarized with the help of existing mathematical models. The main characteristics of the hemodynamic elements, such as the heart and arterial and venous systems, are first described. Distributed models of an arterial network are introduced, and their characteristics are compared with those of lumped parameter models. We also discuss the nonlinear characteristics of the pressure volume relationship in veins. Then the control pathways that participate in feedback mechanisms (baroreceptors and cardiopulmonary receptors) are described to explain the interaction between hemodynamics and autonomic nerve control in the circulation. Based on a set-point model, the computational aspects of reflex control are explained. PMID- 15760488 TI - The effect of e-, i-, and n-nitric oxide synthase inhibition on colonic motility in normal and muscular dystrophy (mdx) mice. AB - To explore the origin of diarrhea or constipation in human Duchenne muscular dystrophy (DMD), the effect of the inhibition of e- , i-, and n-nitric oxide synthase (NOS) on the motility of proximal and distal segments of colon of muscular dystrophy (mdx) and control mice was studied. The frequency of migrating motor complexes (MMC) was higher in the proximal than in the distal segments in mdx colon (0.56 vs. 0.25 cpm) and in the control colon (0.7 vs. 0.25 cpm), and there was no difference when mdx was compared to control segments. High concentrations of NOS inhibitors, including 1,3-PBIT dihydrobromide (1,3-PBIT) and spermine, inhibited MMC. The dose of spermine required to inhibit MMC was lower for the proximal mdx colon than for the distal mdx or control colon. In the presence of tetrodotoxin, spermine (1 mM) and 1,3-PBIT (5 mM) reduced the magnitude of local, rhythmic contractions (LC) paced by the interstitial cells of Cajal (ICC), but 1,3-PBIT (50 microM) increased their magnitude. There was no difference in the effect of spermine and 1,3-PBIT on the LC between mdx and control colon. The results suggest an inhibition of MMC by high concentrations of e-, i-, and n-NOS inhibitors, modulation of ICC activity by e-NOS, and greater susceptibility of MMC to n-NOS inhibition in the mdx proximal than in the control colon, which is very likely because of a deficit in n-NOS in the mdx smooth muscle affecting the MMC pacemaker. A deficit in the effect of mdx smooth muscle n-NOS on an MMC pacemaker may be the origin of diarrhea or constipation in human DMD. PMID- 15760489 TI - Cardiovascular and intravesical pressure responses during natural micturition in conscious rats. AB - Urinary bladder distension is known to influence the cardiovascular system under a pathophysiological condition such as spinal cord injury, hypertension, and arteriosclerosis. A reflex due to bladder distension and/or contraction is considered as one reason for the cardiovascular disturbance associated with micturition. However, it has remained unknown how much intravesical pressure (IVP) rises during micturition in daily life and to what extent mean arterial blood pressure (MAP) and heart rate (HR) respond at that time. To answer these questions, we attempted to examine the direct changes in IVP, MAP, and HR during natural micturition in freely moving conscious rats. IVP increased from the baseline value of 4 +/- 0.2 mmHg to 14 +/- 0.5 mmHg during natural micturition. Although MAP and HR began to increase before micturition, the increases in MAP and HR became significant 1-4 s before its onset. The peak increases in MAP and HR (7 +/- 0.8 mmHg and 14 +/- 3 beats/min, respectively) were delayed by 2 s from the peak IVP. Following an administration of xylocaine into the urinary bladder, the increases in MAP and HR during micturition were significantly blunted to 5 +/ 2 mmHg and 8 +/- 3 beats/min, although IVP increased the same as it did during micturition without xylocaine. Moreover, the relationship between IVP and MAP or HR during natural micturition resembled that between IVP and the vesico cardiovascular reflex responses during isovolumic bladder contraction in anesthetized rats. Therefore it is concluded that natural micturition in freely moving conscious rats accompanies the significant cardiovascular responses despite a limited increase in intravesical pressure, to which a reflex from the urinary bladder may substantially contribute. PMID- 15760490 TI - KR-31378 protects cardiac H9c2 cells from chemical hypoxia-induced cell death via inhibition of JNK/p38 MAPK activation. AB - Using a metabolic inhibition buffer as an ischemic model, we show here that KR 31378, a cardioselective ATP-sensitive potassium channel opener, protects H9c2 cells from chemical hypoxia (CH)-induced cell death. Our previous study showed that CH downregulated caspase activities, but led to differential activation of mitogen-activated protein kinases (MAPKs) in H9c2 cells. The repression of CH induced c-jun N-terminal kinase (JNK)/p38 MAPK activation resulted in partial protection against CH- induced cell death, implying JNK/p38 MAPK's causative role in CH-induced cell death. This study furthers that research and examines if KR 31378's protective effect came from modulating MAPK activity and/or caspase activity in H9c2 cells. Although KR-31378 did not restore downregulated caspase-3 activity, it did block the activation of JNK and p38 MAPK in a dose-dependent manner. Extracellular signal-regulated kinase activity was not recovered by KR 31378 treatment. CH-induced reactive oxygen species (ROS) generation was suppressed by KR-31378. Thus our results indicate that the cardioprotective effect of KR-31378 in CH is due, at least in part, to the differential inhibition of MAPKs. PMID- 15760491 TI - Cooperative effects of exercise and occlusive stimuli on muscular function in low intensity resistance exercise with moderate vascular occlusion. AB - To obtain insight into the relative contributions of exercise and occlusive stimuli to these muscular adaptations, the present study investigated the short- and long-term effects of varied combinations of low-intensity exercise and vascular occlusion. The subjects were separated into 3 groups (n = 6 for each group): low-intensity with vascular occlusion (LIO), low-intensity without vascular occlusion (LI), and vascular occlusion without exercise (VO). LIO and LI groups performed bilateral knee extension exercises in seated positions with an isotonic extension machine. In the LIO group, both sides of the thigh were pressure-occluded at the proximal end by means of a tourniquet during the entire session of exercise (approximately 10 min), whereas only the occlusion with the same pressure and duration was given in the VO group. The mean occlusion pressure was 218 +/- 8.1 mmHg (mean +/- SE). The exercise session consisted of five sets of exercise at an intensity of 10-20% 1RM and was performed twice a week for 8 wk. After the period of exercise training, isometric and isokinetic strengths at all velocities examined increased significantly in the LIO group (p < 0.05), whereas no significant change in strength was seen in the LI and VO groups. The increase in muscular strength in LIO was associated with a significant increase in the cross-sectional area of knee extensor muscles by 10.3 +/- 1.6%. The plasma growth hormone concentration measured 15 min after the session of exercise showed a marked increase only in LIO. The results showed that the low-intensity exercise and occlusive stimuli have cooperative effects in the long-term adaptation of muscle and an acute response to growth hormone. PMID- 15760492 TI - Optical mapping of tachycardia-like excitation evoked by pacing in the isolated rat atrium. AB - Multiple-site optical recording of membrane potential activity using a voltage sensitive dye was employed to monitor action potentials in the rat atrium. Electrical pacing at 1 Hz evoked the events of tachycardia-like excitation (TE). Optical mapping revealed that a coupling of spontaneous and evoked action potentials resulted in the initiation of TE. PMID- 15760493 TI - Some consequences of the global obesity epidemic. PMID- 15760494 TI - Contribution of bariatric surgery to the comprehension of morbid obesity. AB - Convinced that morbid obesity was not due to food excess but rather to a metabolic disorder, we searched in the literature for data in favor of a metabolic disorder. We have found evidence in support of the thesis that the cause of morbid obesity is the inability to burn excessive caloric intake normally. It would involve the difficulty to increase heat with the amount of calories taken, which would be faulty and force fat deposition. This mechanism called dietinduced thermogenesis (DIT) allows the dispersion by heat of excessive calories to obtain energy balance. Results from bariatric surgery and particularly biliopancreatic diversion (BPD) give further support to this thesis. BPD would improve heat production to a meal (DIT) by one of these mechanisms: increased insulin sensitivity, change in intestinal hormone secretion, or chronic lipid malabsorption. Available results show that surgery, to be efficient, must change the physiology and not solely decrease food intake. PMID- 15760495 TI - Relationship between single nucleotide polymorphisms in leptin, IL6 and adiponectin genes and their circulating product in morbidly obese subjects before and after gastric banding surgery. AB - BACKGROUND: Certain adipose-produced signals are secreted in proportion to body fat mass and are involved in regulation of the energy metabolism of the whole body. Leptin, IL6 and adiponectin can be considered as adiposity signals. Several Single Nucleotide Polymorphisms (SNPs) in genes encoding for these molecules are known to influence their concentration in situations of stable weight. We hypothesized that polymorphism effects could be better detected in a situation of negative energy balance and that modified concentrations of adiposity signal genes could change the dynamics of weight gain in obese subjects. METHODS: 65 obese patients undergoing gastric banding surgery were genotyped for LEP+19A-->G, LEP-2548G-->C, IL6-174G-->C, APM1-11377C-->G and PM1-11391G-->A common SNPs. BMI and concentrations of leptin, IL6 and adiponectin were measured before surgery and after 1 year. RESULTS: All SNPs except IL6-174G-->C SNP were associated with modifications of the circulating concentrations of signals produced by adipose tissue at baseline. During weight loss, variant genotype carriers of LEP -2548 and +19 SNPs were characterized by a trend towards less decrease in circulating leptin. Weight loss was associated with an increase in IL6 concentration (16.9%+/ 12.2) in the IL6-174 C/C genotype carriers, whereas the C/G or G/G genotypes carriers showed a decrease in IL6 (19.9%+/-5.2, P=0.001). CONCLUSION: We observed that the SNPs studied could modulate the concentration of adiposity signals not only at baseline but also during weight loss. Such variations may be sensed by the homeostatic feedback system that controls energy balance and may in turn contribute to some disturbances in weight regulation. PMID- 15760496 TI - The cost effectiveness of laparoscopic versus open gastric bypass surgery. AB - BACKGROUND: Over the last decade, laparoscopic gastric bypass (LGBP) has been proven to be a safe and well-tolerated approach to the Roux-en-Y gastric bypass, despite its increased cost when compared to the open approach (OGBP). This increased expense has led many to question whether LGBP is a cost effective alternative to OGBP. The aim of this study is to determine which approach is most cost effective, considering costs associated with the operation itself, perioperative complications, and income lost during convalescence. METHODS: A PubMed search of the National Library of Medicine online journal database was conducted. Studies that met predetermined criteria for selection were included in the analyses of patient demographics, perioperative complications, length of hospital stay, excess weight loss, and time to recovery. Data on 6,425 OGBP and 5,867 LGBP patients were used to compare the outcomes associated with each approach. RESULTS: Significant differences were found in the perioperative complication profiles, time to recovery, and overall expense of the two approaches. OGBP was associated with an increased incidence of major perioperative complications, especially extraintestinal complications, and greater perioperative mortality. LGBP was associated with shorter hospital stays, increased incidence of intestinal complications, and a 2.25% incidence of conversion to OGBP. Patient demographics and percent excess weight loss (%EWL) at 3 years follow-up were found to be similar with both OGBP and LGBP. CONCLUSION: LGBP is a cost effective alternative to OGBP for surgical weight loss. Despite the increased cost of LGBP, patients suffer fewer expensive and lifethreatening perioperative complications. PMID- 15760497 TI - Laparoscopic Roux-en-Y gastric bypass with 2-metre long biliopancreatic limb for morbid obesity: technique and experience with the first 150 patients. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass (RYGBP) is being performed widely as a treatment of choice for morbid obesity. We present our method and experience with the first 150 consecutive cases of laparoscopic RYGBP with a 2-m long biliopancreatic limb (BP-limb). METHODS: Between November 2001 and November 2003, a prospective analysis of 150 patients was performed identifying technical success and complications. Before surgery, patients underwent a strict multidisciplinary behavioral program. At operation the stomach was transected proximally with a linear stapler (60-mm, Endo-GIA) to create a prolongation of the esophagus (gastric tube) along the lesser curvature, resulting in a 40-50 ml pouch. Two meters of the proximal jejunum were bypassed (BP-limb), creating an antecolic Roux-en-Y gastro-jejunostomy to the posterior wall of the gastric tube using a 45-mm linear Endo-GIA stapler. The entero-anastomosis was created 50 cm below the gastro-jejunostomy, also with a 45-mm linear Endo-GIA. RESULTS: Mean BMI was 50.0, and 78% of patients were females. With 100% follow-up, we found an EWL of 50% 6 months after surgery, gradually rising to 80% after 18 months. The mean operating time was 116 min for the first 50 cases and decreased to 82 min for the last 50 cases. Intestinal leakage occurred in 5 patients (3%) and bleeding in 5 (3%). Most of these complications occurred in the first 50 cases, and all but one were treated successfully with an early laparoscopic re operation. Marginal ulcers were found in 16.6% of patients. No internal hernias have occurred. CONCLUSION: The operation demands advanced laparoscopic skills, but technically it is relatively simple and has an acceptable complication rate. Short-term results regarding excess weight loss are at least comparable to the RYGBP with a long alimentary limb. PMID- 15760498 TI - Laparoscopic Roux-en-Y gastric bypass in patients with BMI <50: a prospective randomized trial comparing short and long limb lengths. AB - BACKGROUND: It has been shown that long limb gastric bypass in the super-obese (BMI >50) results in increased weight loss in comparison with conventional gastric bypass. The purpose of this study was to compare the effect of short and long limb lengths in patients with BMI<50. METHODS: 48 patients with BMI <50 (46 females / 2 males, mean age 35+/-9.6 years) were prospectively randomized to either a short limb (biliopancreatic limb = 50 cm, alimentary limb = 100 cm) or long limb (biliopancreatic limb = 100 cm, alimentary limb = 150 cm) laparoscopic Roux-en-Y gastric bypass (LRYGBP). In all patients, a 25-mm EEA was used to fashion the gastrojejunostomy and the Roux limb was positioned in an antecolic, antegastric location. Limb lengths were precisely measured in all cases. RESULTS: There was no difference in demographic data, preoperative BMI, presence of co morbidities, or duration of surgery. The overall complication rate was not different between the 2 groups; however, the incidence of internal hernias was significantly higher in the long limb group (0 vs 4, P=0.029). The length of hospital stay was longer for the short limb group compared to the long limb group (3.1 vs 2.2 respectively, P=0.004). When comparing the short limb to the long limb patients, the BMI decreased equally in both groups at the following time intervals: preoperative (44.6 vs 44.9), 3 weeks (40.3 vs 40.9), 3 months (35.5 vs 35.2), 6 months (31.2 vs 31.8), and 12 months (27.7 vs 28.3). There were no significant nutritional deficiencies in either group. CONCLUSIONS: In patients with BMI <50 undergoing LRYGBP, increasing the length of the Roux limb does not improve weight loss and may lead to a higher incidence of internal hernias. PMID- 15760499 TI - General anesthesia, bariatric surgery, and the BIS monitor. PMID- 15760500 TI - Beneficial effects of humidified, warmed carbon dioxide insufflation during laparoscopic bariatric surgery: a randomized clinical trial. AB - BACKGROUND: Recent data has shown that the use of warmed, humidified carbon dioxide (CO2) insufflation during laparoscopic surgery may be associated with better outcomes. METHODS: We performed a randomized, doubleblind, prospective controlled clinical trial of 30 patients undergoing laparoscopic Roux-en-Y gastric bypass (LRYGBP). Patients were randomized into 2 groups. The first group (group 1, n=15) received standard (dry, room temperature) CO2 for insufflation during the surgery, while the second group (group 2, n=15) received warmed (35 degrees C) and humidified (95%) CO2. Patients received postoperative analgesia from morphine delivered via a patient-controlled analgesia (PCA) pump. Pain scores (on a scale of 0 to 10, 0 being no pain and 10 being the worst pain) were measured postoperatively at 3 h, 6 h, 1 day and 2 days. The amount of morphine that was delivered through the PCA was also measured at the same time intervals. Operating-room (OR) time, core temperature, and total hospital length of stay were documented. RESULTS: Postoperative pain as documented by pain scores and narcotic usage were not statistically different in the 2 groups. We demonstrated a statistically significant difference (mean+/-SD) in OR time (76+/-16 min vs 101+/-34 min, P=0.02), total hospital length of stay (3.2+/-.4 days vs 4.0+/-.9 days, P=0.01) and end-of-case core temperature (36.2+/-.5 degrees C vs 35.7+/-.6 degrees C, P=0.02) in group 2 compared with group 1. CONCLUSION: The use of warmed, humidified CO2 insufflation in bariatric patients undergoing LRYGBP was not associated with any significant benefit with regards to postoperative pain. PMID- 15760501 TI - Impact of a warm gas insufflation on operating-room ergonometrics during laparoscopic gastric bypass: a pilot study. AB - BACKGROUND: With the use of various laparoscopic instruments, the work of operating-room (OR) personnel has increased significantly. The impact of warm gas insufflation on the ergonometrics of the OR was studied, using one of the most involved laparoscopic surgical procedures, Roux-en-Y gastric bypass (RYGBP) for morbid obesity, to assess if use of warm gas insufflation decreases the work of the OR personnel. METHODS: 20 patients between August 2003 and January 2004 (6 months) were divided into 2 groups. 10 patients with age 50+/-10 years and BMI 48+/-8 underwent laparoscopic RYGBP using a warmed CO2 insufflator (WI). These results were compared to 10 patients with age 53+/-15 years and BMI 51+/-7 using a non-warmed CO2) insufflator (NWI). Total time of surgery (TOS), time spent cleaning the laparoscope (TCS), time spent changing warm saline (TWS), time spent using anti-fog (TAF), and time the circulating nurse was involved in these activities (TN) were compared. Statistical analysis used a two-sample, Student t test with unequal variances. RESULTS: The 2 bariatric populations were almost similar in age and BMI. TCS (P<0.0003), TWS (P<0.0001) and TN (P<0.0002) took significantly less time in the WI group, while TOS and TAF were similar. CONCLUSION: Use of warmed CO2 insufflation had a significant impact on TCS, TWS and TN. This impacts the ergonometrics of the OR, allowing more time for the personnel and surgeons to concentrate on the surgery. PMID- 15760502 TI - Is heated and humidified gas necessary during laparoscopic gastric bypass? PMID- 15760503 TI - Laparoscopic gastric bypass versus laparoscopic adjustable gastric banding in the super-obese: a comparative study of 290 patients. AB - BACKGROUND: Controversy exists regarding the best surgical treatment for super obesity (BMI >50 kg/m2). The two most common bariatric procedures performed worldwide are laparoscopic adjustable gastric banding (LAGB) and laparoscopic Roux-en-Y gastric bypass (LRYGBP). We undertook a retrospective single-center study to compare the safety and efficacy of these two operations in super-obese patients. METHODS: 290 super-obese patients underwent laparoscopic bariatric surgery: 179 LAGB and 111 LRYGBP. RESULTS: There were one death in both groups. The early complication rate was higher in the LAGB group (10% vs 2.8%, P<0.01). Late complication rate was higher in the LAGB group (26% vs 15.3%, P<0.05). Operating time and hospital stay were significantly higher in the LRYGBP group. LRYGBP had significantly better excess weight loss than LAGB (63% vs 41% at 1 year, and 73% vs 46% at 2 years), as well as lower BMI than LAGB (35 vs 41 at 18 months). CONCLUSION: LRYGBP results in significantly greater weight loss than LAGB in super-obese patients, but is associated with a higher early complication rate. PMID- 15760504 TI - Laparoscopic adjustable gastric banding: early experience in 400 consecutive patients in the USA. AB - BACKGROUND: Early experience with 400 consecutive patients who underwent laparoscopic adjustable gastric banding (LAGB) is reported. METHODS: From Nov 2002 to Aug 2004, prospective data were collected on 400 consecutive LAGB patients and evaluated retrospectively. RESULTS: There were 354 (88.5%) females and 46 males (11.5%), with mean age 43.6 years and mean BMI 46.2 kg/m2. For outpatients (freestanding ambulatory surgery center), mean OR time was 55.4 min in 208 patients (52%), compared to mean inpatient OR time of 70.5 min in 192 patients. Inpatients had a higher BMI (48.2 +/- 9.3 SD) than outpatients (43.9 +/ 5.7 SD) (P<0.0001). Complications occurred in 35 patients (8.8%). These consisted of 9 slipped bands (2.3%) that were surgically repositioned, 6 port problems (1.5%) that were successfully repaired, 17 patients with temporary stoma occlusion (4.3%) that spontaneously resolved, and 2 bowel perforations (0.5%) that required surgical repair and band removal. One patient died of pneumonia 2 weeks after an uneventful procedure. Average 1-year percent excess weight loss (%EWL) in 138 patients was 48.2%. Patients who had < or =50 kg initial excess weight (n=37, 27%) had a significantly higher %EWL (55.2%) at 1 year than patients who had >50 kg initial excess weight (P=0.0011). CONCLUSIONS: LAGB has been safe and effective thus far for the surgical treatment of morbid obesity, and can be performed as an outpatient in select patients. PMID- 15760505 TI - Nutritional behavior as a predictor of early success after vertical gastroplasty. AB - BACKGROUND: Patients' nutritional habits are seldom taken in account in planning surgery for clinically severe obesity. Our proposed hypothesis is that the patient's nutritional behavior may influence the outcome of bariatric surgery. METHODS: The impact of nutritional behavior on the postoperative weight-loss was evaluated before and after bariatric surgery. A 6-month prospective consecutive case study was carried out on patients undergoing a Silastic ring vertical gastroplasty (SRVG). Patients were interviewed and examined before and at 1, 3 and 6 months after surgery. Demographic and clinical data were collected from the patients' medical charts. Nutritional data collected from a self-filled questionnaire included information on hunger and satiety perception, nutritional behavior (intake, eating habits and maximum consistency of consumed food) and concomitant symptoms. RESULTS: The sample included 69 patients: 56 were women (81%); average age was 32 years (range 18 50). Average preoperative BMI was 43.4 +/- 5.3 kg/m2 (range 35-58). 6 months after surgery, BMI was 30.3 +/- 3.8 kg/m2 (range 21-42). Weight loss forecast models showed a statistically significant role of factors related to: anthropometrical preoperative data, hunger perception, prevalence of oral mucosal sore, and nutritional behavior. CONCLUSION: The short nutrition outcomes after gastric restrictive surgery were looked at, with their impact on weight-loss success. The Eating Status concept should be part of a systematic profiling of morbidly obese patients for preoperative nutritional behavior and postoperative nutritional education, to achieve the best comprehensive treatment in regard to weight loss and quality of life. PMID- 15760506 TI - Implantable gastric stimulation inhibits gastric motility via sympathetic pathway in dogs. AB - BACKGROUND: The aim of this study was to investigate the effect of implantable gastric stimulation (IGS) on gastric antral contractions and the involvement of the sympathetic pathway. METHODS: The study was performed in 5 postprandial sessions in 8 dogs chronically implanted with stimulation electrodes and a gastric cannula: a) IGS via lesser curvature; b) IGS via antrum; c) and d) same as a) and b) but IGS initiated 1 hr before the meal; e) same as a) but with guanethidine. RESULTS: It was found that: 1) IGS significantly inhibited postprandial antral contractions assessed by manometry, and no significant difference was noted in the effect between the two stimulation sites; 2) IGS initiated 1 hr before the meal was more potent than that initiated 30 minutes after the meal; 3) the inhibitory effect of IGS on postprandial antral motility was completely blocked by guanethidine. CONCLUSION: Acute IGS inhibits postprandial antral contractions, and this inhibitory effect is mediated via the sympathetic pathway. PMID- 15760507 TI - Intragastric balloon for obesity: a retrospective evaluation of tolerance and efficacy. AB - BACKGROUND: The intragastric balloon may be used for weight reduction for mild or moderate obesity, or for preoperative weight loss for super-obesity. The authors retrospectively evaluated the tolerance and efficacy of the BioEnterics Intragastric Balloon (BIB). METHODS: From October 2002 to July 2004, intragastric balloons were placed, under endoscopic control, in 44 patients (mean BMI 45 kg/m2 , mean age 31 years). The balloons were filled with 500-600 mL of normal saline. Removal was recommended for 6 months after balloon insertion. RESULTS: 6 patients (13.6%) were lost to follow-up, 7 super-obese patients underwent LAGB at our hospital, and 2 patients had the BIB procedure performed twice. Balloon placement was uneventful. Removal was performed endoscopically in 38 patients under conscious sedation with anesthesiological assistance (2 patients had the BIB removed under general anesthesia). No cases of tracheal aspiration or spontaneous balloon evacuation were encountered. Sideeffects were vomiting during the 1st week (77.2%), occasional vomiting for >3 weeks (11.3%), hypokalemia (6.8%), functional renal insufficiency (4.5%), abdominal pain (15.9%), and gastroesophageal reflux (6.8%). There was 1 gastric perforation (treated laparoscopically after removal of the BIB), 1 gastric ulcer, 4 cases of intolerance (1 of these elected to have LAGB), and 1 died (from other medical conditions). Mean excess weight loss was 13 kg (33 kg in the super-obese). CONCLUSIONS: The BIB appears to be safe provided that it is removed within the specified 6 months. Surveillance is necessary. It was efficient in reducing weight in patients with mild or moderate obesity and as preoperative treatment for super-obese patients to reduce the surgical risk before LAGB. PMID- 15760508 TI - Surgery for morbid obesity: selection of operation based on evidence from literature review. AB - BACKGROUND: Bariatric surgery is the therapy for morbid obesity. There are a number of surgical procedures, which are performed by open surgery (OS) and more recently also by laparoscopy (LS). The objective of this study was to consider the evidence for the best bariatric surgical options. METHODS: Systematic review of the literature was conducted. Morbid obesity studies published between 1990 and 2002 were analyzed. MEDLINE, LiLACS and COCHRANE databases were used, utilizing MeSH terms and free words. Selected studies were analyzed using a specially designed score of methodological quality, to analyze and compare studies. This validated scale is composed by 3 items, and the final score may range between 6 and 36 points. Means, medians and weighted means were calculated, and a comparative analysis by therapy was performed using median 95% confidence intervals (CI). Number of treated patients, reduction in body mass index (BMI), reduction in co-morbidity, %EWL, morbidity and mortality, hospital stay, follow up, success and failure of operations, and methodological quality were analyzed. RESULTS: 283 related articles were considered. Only 31 of them had selection criteria (these include 5,216 patients operated by OS and 3,230 by LS). Operative mortality was 0.0% for OS and 0.4% for LS. At 36 months, OS techniques show reduction in BMI, %EWL and reductions in co-morbidity of 30.9%, 61.9% and 74.1% respectively. At 36 months, LS techniques show reduction in BMI, %EWL and reduction in comorbidity of 23.7%, 55.9% and 70.9%. Hospital stay was 3.8 days for LS and 7 for OS. At 2-year follow-up, morbidity was 14.8% for LS and 16.7% for OS, and reoperations were 17.7% for LS and 11.3% for OS. Median score for methodological quality was 13 for OS and 11 for LS. CONCLUSIONS: Methodological quality of primary studies to 2002 has been poor. PMID- 15760509 TI - Choosing an operation for weight control, and the transected banded gastric bypass. AB - Obesity and particularly morbid obesity is a lifelong problem that currently cannot be cured but can be controlled. Attempted control of obesity non surgically results in 98% recividism. Weight loss is readily attainable, but weight loss maintenance is recalcitrant. Surgery currently provides the only long term control of obesity. Surgery at best is a tool that the patient can use to effect the weight loss and weight loss maintenance. We have celebrated the golden anniversary of bariatric surgery in 2004. Obesity surgery is thus a relatively young field which is evolving. Operations currently used for the treatment of obesity fall into 3 categories: 1) restrictive operations such as vertical banded gastroplasty, silastic ring gastroplasty and gastric banding; 2) malabsorptive operations which include all the variations of the intestinal bypass; and 3) combined operations which utilize both restriction and malabsorption which include all the variations of short-limb gastric bypass, long-limb or distal gastric bypass and biliopancreatic diversion. The choice of the operation will be guided by the extent of the patient's obesity, the age of the patient, other co morbid conditions of the patient, the cost of the operation, the patient's choice, and the surgeon's choice based on training, experience and geographical location. First and foremost, the operation chosen should be effective in causing weight loss and providing long-term weight loss maintenance with acceptable morbidity and mortality. Recommendations are made for choosing an operation for weight control based on effectiveness and safety. PMID- 15760510 TI - A simple novel technique for intragastric balloon retrieval. AB - We present a novel technique for the retrieval of intragastric balloons used in the treatment of patients with morbid obesity. Although specifically-designed instruments are available for this purpose, this simple technique makes use of instruments that are readily available in an endoscopy suite equipped for simple therapeutic procedures. PMID- 15760511 TI - Practice makes perfect? Patient response to a prebariatric surgery behavior modification program. AB - BACKGROUND: Although bariatric surgery has been identified as an effective treatment for weight loss in the morbidly obese, some patients regain weight postoperatively. A pre-treatment program focused on changing lifestyle behaviors could facilitate improved post-surgical outcomes. The purpose of this study was to determine patient satisfaction and perceived usefulness of a preoperative program. METHOD: Participants completed a bariatric surgery program that incorporated a 6-week behavior modification component before the surgery date. A selfreport questionnaire was sent to individuals who had completed the program. The questionnaire assessed demographics, satisfaction with the pre-surgery program, and patients' perceived usefulness of the program. RESULTS: Of 124 questionnaires sent, 70 were returned (56%). Participants had a mean age of 46.3 years. The majority of participants were Caucasian (97.1%), female (78.6%), and employed (68.6%). Mean preoperative BMI was 55.3; mean postoperative BMI was 36.3. Participants were 1 year post-surgery (mean 48.8 weeks). Patients were very satisfied with the overall program (mean Likert score 4.51), as well as with the behavior modification groups (mean 4.44). They found the groups to be very useful in making postoperative changes (mean 4.45). Patient satisfaction and perceived usefulness were not significantly correlated with the degree of weight lost (r=.05, P=.69; r=.05, P=.71). CONCLUSIONS: Patients found the preoperative behavior modification program useful in helping them make the necessary post surgical lifestyle changes. PMID- 15760512 TI - Band slippage--a potentially life-threatening complication after laparoscopic adjustable gastric banding. AB - BACKGROUND: Although gastric bands are safe and effective devices, severe late complications may develop in rare cases. PATIENTS: 3 patients were treated for complete dysphagia after slippage of gastric bands. 2 of the patients were admitted for severe dehydration, 1 of whom developed cerebral venous infarction. Ischemia of the gastric pouch occurred in 1 patient. RESULTS: All 3 patients survived after successful medical therapy and surgical removal of the bands. Bariatric reoperations were performed in 2 patients (gastric sleeve resection, gastric bypass). CONCLUSION: Complete dysphagia on the basis of band slippage represents a life-threatening acute event, which may occur even years after implantation. Patients and doctors should be informed about this long-term risk of gastric banding. PMID- 15760513 TI - Renal failure, glomerulonephritis and morbid obesity: improvement after rapid weight loss following laparoscopic gastric bypass. AB - There is experimental evidence but very few human studies that suggest a role for obesity in the formation and progression of some glomerular lesions. We report the case of a morbidly obese male with hematuria and proteinuria that was subsequently diagnosed with renal failure which required dialysis. Histological findings of the renal biopsy performed during a laparoscopic gastric bypass are presented. His renal failure resolved with the weight loss. PMID- 15760516 TI - [Understanding the nature of medicine from the development of disaster medicine]. PMID- 15760514 TI - Universal laparoscopic adjustable gastric banding? PMID- 15760517 TI - [Risk management and countermeasures for comprehensive hospital]. PMID- 15760518 TI - [Guidance of therapy guidelines 2004 ACC/AHA for ST elevation acute myocardial infarction complicated by arrhythmia]. PMID- 15760520 TI - [Clinical efficacy and safety of emergent percutaneous coronary intervention for vein-grafts in the patients with acute myocardial infarction: comparison between age groups]. AB - OBJECTIVE: To evaluate the efficacy and the safety of emergent primary percutaneous coronary intervention (PCI) in the saphenous vein bypass graft (SVBG) of acute myocardial infarction (AMI), and compare the results between aged -patients with non-aged patients. METHODS: Three hundred and nine consecutive AMI patients with culprit SVBG vessels, were analysed, including aged patients 213 cases(>or=70 years old), non-aged patients 96 cases(<70 years old), underwent the emergent primary PCI after confirmed below TIMI III perfusion(TIMI 0-TIMI II) in coronary angiographies. The immediate results and in-hospital outcomes were compared between two groups. RESULTS: Procedural successful rate, re-occlusion rate, and emergency re-CABG had no significant differences between two groups. The rate of slow-flow/no-reflow and in-hospital mortality rate were significantly higher in elderly group (19.7% vs 10.4%, 9.4% vs 4.2%, both P<0.05), with no difference in the rate of the using of distal protection devices between two groups. The comparison of the rate of direct stenting in slow-flow/no-reflow subgroup with normal-flow subgroup, had not showed statistic difference (73.5% vs 67.3%, P>0.05). There was no statistic difference of heavy hemorrhage between two different age groups. CONCLUSION: The primary PCI for the elderly AMI patients with infarction-related SVBG vessels, has higher risks in slow-flow/no-reflow and the mortality, even with using the distal protection devises and direct stents implantation. PMID- 15760521 TI - [Risk factors of sudden death and death from arrhythmia in patients with non-ST elevation acute coronary syndromes in China]. AB - OBJECTIVE: To analyze the relationship between sudden death and death from arrhythmia and multiple risk factors in patients with non-ST elevation acute coronary syndromes in two years of follow-up in China. METHODS: This study was a part of an international multicentre registry-Organization to Assess Strategies for Ischemic Syndromes (OASIS). The patients admitted to the hospital with non-ST elevation acute coronary syndrome were enrolled. No particular intervention was given for the treatment. All patients had been followed-up for two years. The patients' clinical characteristics, therapeutic regimes and major events during hospitalization and two years' follow-up period were recorded by filling in Case Report Forms according to the protocol offered by Canadian Cardiovascular Collaboration. Cox regression model was used to analyze the association with the most common causes of death and multiple factors recorded. RESULTS: From April 1999 to December 2001, 2294 cases were enrolled in 38 hospitals in China nationwide. Among them 2188 patients two years follow-up was accomplished. The mean age of the patients was (62.8+/-8.1) years. Male gender was dominant (62.3%). The clinical diagnosis at admission was unstable angina in 88.5% of the patients and non Q-wave myocardial infarction(MI) in the remaining 11.5%. The mortality was 7.6% with total deaths of 174 by the end of 24-month follow up. The most common cause of death was severe arrhythmias or sudden death (92 cases, 52.9%). More than 70 factors were analyzed by Cox regression model in order to determine which were the predominant factors of death. Major risk factors that predisposed to death were: number of episodes of MI during follow-up period, re MI within 24 hours during hospitalization, heart failure during hospitalization, previous history of MI, recurrent angina pectoris during hospitalization, the duration of hospitalization, and patient age. Protective factors that reduced the chance of death were: the frequency of using nitrate, frequency of taking anti platelet medicine or beta-blocker during follow-up period. CONCLUSION: In China, the most common cause of death in patients with non-ST elevation acute coronary syndromes is severe arrhythmias or sudden death, and it is related with the severity of coronary artery disease and the age of patients in majority of cases. Some factors that influence survival are similar to those established by previous evidence based medicine. PMID- 15760522 TI - [Characteristics in the feature of 3H-Ryanodine binding to cardiomyocyte nuclei in reperfusion injury in rat]. AB - OBJECTIVE: To investigate the changes in binding features of (3)H -Ryanodine cardiac myocytic nuclei in reperfusion injury in the rat, and the effects of phosphorylation regulation on the binding characteristics. METHODS: Healthy male Wistar rats were randomly divided into ischemia/reperfusion injury (IRI) group and sham-operation group. IRI model was reproduced by ligating the left main coronary artery for 30 minutes followed by reperfusion of the heart for 3 hours, while in the sham-operation group the animals underwent a thoracotomy only for 3.5 hours. Cardiac myocytic nuclei were isolated by sucrose density gradient centrifugation. The maximum binding capacity (Bmax) and the dissociating ratio (Kd) of Ryanodine receptors (RyRs) were measured by radioligand binding analysis as well as Scatchard plot. RESULTS: There was a high affinity of RyRs to bind with (3)H-Ryanodine on the nuclear wall of rat cardiac myocytic nucleus. Compared with that of the sham-operation group, Bmax of RyRs of IRI cardiac myocytic nuclei was decreased by 29% (P<0.01), but there was no difference in Kd between two groups(P>0.05). With phosphorylation by activating the endogenous protein kinase C(PKC) with phorbol myristate acetate (PMA)+phosphatidyl serine(PS), Bmax of both sham and IRI groups were increased markedly (P<0.01), but in the latter group it was less increased (P<0.01). With phosphorylation by Ca(2+)-calmodulin (CaM), Bmax was decreased in both sham-operation and IRI groups (both P<0.05), but was less decreased in the latter(P<0.01). However, both PMA+PS and Ca(2+)-CaM did not change the Kd of nuclear RyR in either group (both P>0.05). CONCLUSION: After IRI, Bmax of (3)H -Ryanodine of cardiac myocytic nuclei is decreased and the impact of PMA+PS and Ca(2+)-CaM on the Bmax is impaired, but the affinity of (3)H -Ryanodine to cardiac myocytic nuclei is not altered under above circumstances. PMID- 15760524 TI - [Role of mitogen - activated protein kinases signal pathway in cardiomyocyte injury induced by serum after hypoxia and burn injury]. AB - OBJECTIVE: To observe the activation and explore the role of three major mitogen activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), p38 kinase and c-Jun NH(2)-terminal protein kinase (JNK), in cardiomyocytes injury induced by serum after hypoxia and burn injury. METHODS: Phosphorylation of the three major MAPKs in primary cultured neonatal rat cardiomyocytes were determined by Western blotting. Contents of released lactate dehydrogenases (LDH) and death-rate of myocytes treated with serum after hypoxia and burn injury, SB203580+hypoxia and burn serum, PD98059+hypoxia and burn serum were observed respectively. RESULTS: Exposing rat neonatal cardiomyocytes to hypoxia and burn serum resulted in a rapid and prolonged activation of p38 kinase and ERK. Phosphorylation degree of p38 kinase, ERK1/2 was increased. Myocytes treated with SB203580 (10 micromol/L), a selective inhibitor of p38 kinase, resulted in a significant decline in LDH leakage leaking and cell death. However, with pretreatment of cell with PD98059(25 micromol/L), an inhibitor of ERK, LDH leakage and cell death were increased. CONCLUSION: Serum obtained after hypoxia and burn injury activate p38 kinase and ERK, but not JNK, in cardiomyocytes. p38 kinase pathway might play a role in mediating cardiomyocytes injury, whereas ERK plays a protective role. PMID- 15760525 TI - [Protective effect of phenolic alkaloids from Menispermum dauricum on myocardial cerebral ischemia/reperfusion injury in rabbits]. AB - OBJECTIVE: To explore the mechanism underlying the effects of phenolic alkaloids abstracted from Menispermum dauricum(PAMd) on concurrent myocardial and cerebral ischemia/reperfusion injury in rabbits. METHODS: Both left anterior descending coronary artery and bilateral carotid arteries were ligated to induce myocardial and cerebral ischemia/reperfusion injury at the same time in rabbits. At 30 minutes after ischemia, the occlusion was released to allow return of circulation, and 2 ml of blood was collected from the femoral vein 10 minutes before ischemia, 1, 10, 30 minutes after ischemia, and 1,10, 30, 60, 120, 180, 240 minutes after reperfusion. All rabbits were sacrificed at the end of reperfusion, and the left ventricle, hippocampus, cortex, and cerebellum were removed for examination. Malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. RESULTS: At 10 minutes after reperfusion, MDA content in serum was significantly higher and SOD activity was significantly lower in ischemia/reperfusion group than those of control group (both P<0.05). In rabbits with administration of PAMd, MDA content was lower and SOD activity was higher in serum compared with ischemia/reperfusion group(P<0.05). Similar results as in the blood in regard to MDA content and SOD activity were obtained in tissues. CONCLUSION: PAMd could attenuate the injury induced by lipid peroxidation and enhance the activity of SOD, thus it might play a protective role in concurrent myocardial-cerebral ischemia/reperfusion injury. PMID- 15760528 TI - [Changes in content of plasma brain natriuretic peptide in dilated cardiomyopathy in rabbits]. AB - OBJECTIVE: To determine the changes in plasma brain natriuretic peptide (BNP) in adriamycin-induced dilated cardiomyopathy (DCM) in rabbit, and to evaluated the significance. METHODS: Twenty-two rabbits were randomly divided into control group (n=10) and model group(n=12). The DCM model was reproduced by injecting adriamycin via ear vein for 8 weeks. Echocardiogram was performed and plasma BNP were measured before administration, and at 8 th and 11 th week after the challenge. Indexes of hemodynamics and pathological changes were observed. RESULTS: Indexes of echocardiogram and hemodynamics of model group were consistent with pathologic changes of DCM. Plasma levels of BNP of the model group were increased significantly after administration of the drug(all P<0.01), though the values before the drug administration were approximately the same as in the control group. Plasma BNP levels were significantly higher in DCM group at the 11 th week than at the 8 th week (P<0.05). Plasma levels of BNP were positively correlated with and left ventricular end-diastolic volume(LVEDV), left ventricular end-systolic volume (LVESV), left ventricular end-diastolic pressure(LVEDP) and negatively correlated with left ventricular systolic pressure(LVSP), and left ventricular ejection fraction (LVEF). CONCLUSION: Administration of intravenous adriamycin to rabbits results in DCM which in suitable for the conduction of research of neuroendocrine abnormality of heart. Overload of the left ventricle and increasing tension of left ventricular wall are key factors for regulating BNP excretion. Plasma BNP level is a good marker for evaluating degree of severity of cardiac function in DCM. PMID- 15760526 TI - [Cardioprotective effects of K(ATP) channel opener nicorandil during ischemia/ reperfusion in dogs]. AB - OBJECTIVE: To further confirm the effect of nicorandil in reducing the area of myocardial infarct is through the mediation of activation of the KATP channel but not by its nitrate-like properties, and to determine whether the protective effect is the same or not when the drug is either given immediate after infarction or after reperfusion. METHODS: Thirty-five dogs were randomly divided into five groups as follows. Ischemia/reperfusion (IR) group: the dogs were subjected to 90 minutes of left anterior descending coronary artery(LAD) occlusion followed by 120 minutes reperfusion. Pre-nicorandil (PNIC) group: nicorandil(NIC) 100 microg/kg was administrated by intravenous injection 10 minutes before occlusion, followed by infusion of 10 microg x kg(-1) x min (-1) of the drug till the end of reperfusion. Ischemia nicorandil(INIC) group: NIC 100 microg/kg was administered by intravenous injection 15 minutes after occlusion and 10 microg x kg(-1) x min (-1) of drug intravenously till the end of reperfusion. In the Onset reperfusion treated with nicorandil(RNIC) group: NIC 100 microg/kg was administered intravenously at the onset of reperfusion followed by 10 microg x kg(-1) x min (-1)of drug intravenously up to the end of reperfusion. Glibenclamide(GLIB)+INIC group: before NIC was administered, dogs were pretreated with GLIB 0.3 mg/kg for 10 minutes, and other treatment was the same as INIC group. Hemodynamics data were determined as baseline, 60 minutes post-occlusion, and 120 minutes post-reperfusion. By using 2, 3, 5 triphenyltetrazolium chloride (TTC) staining, the infarct areas were analyzed with image analyzer. RESULTS: The results showed that at 60 minutes post occlusion, cardiac output (CO) was reduced in every group compared with baseline (all P<0.01), CO value recovered at 120 minutes after reperfusion in both PNIC and INIC group (P>0.05). There were no significant differences in heart rate (HR), mean artery pressure(MAP), mean pulmonary artery pressure(MPAP), pulmonary capillary wedge pressure(PCWP) values among all the groups. A marked reduction in the infarct area was found in PNIC group and INIC group (P<0.01) compared with IR group. Administration of GLIB before INIC shows to have no protective effect (P>0.05). CONCLUSION: Our study shows that nicorandil which is a K ATP channel activator, could mimic the effect of ischemic pre-conditioning of the myocardium, by reducing the area of myocardial infarct. PMID- 15760530 TI - [Change in plasma free 15 - F2t - isoprostane concentrations in smoking patients undergoing coronary artery bypass grafting and its clinical significance]. AB - OBJECTIVE: To investigate the change in plasma free 15-F2t-isoprostane fn(15 F(2t)-IsoP) concentration and its relationship with postoperative cardiac function of smoking patients who underwent coronary artery bypass grafting (CABG). METHODS: Sixty patients with coronary artery heart disease were divided into smoking group [n=30, age 54-69 years old, American Heart Association (AHA)II III] and non-smoking group(n=30, age 58-70 years old, ASA II-III). All the patients were scheduled for CABG under combination anesthesia with isoflurane and fentanyl and conventional cardiopulmonary bypass. Blood samples were drawn from the central vein to measure 15-F(2t)-IsoP at the following time points: before operation (T0); 30 minutes after aortic clamping for cardiopulmonary bypass(T1); 10 and 30 minutes after aortic declamping(T2,T3). RESULTS: There was no statistical difference in 15-F(2t)-IsoP between two groups at T0, but contents of 15-F(2t)-IsoP in the two groups was increased markedly at T1, compared to those at T0 (P<0.05 and P<0.01 for smoking and non-smoking groups, respectively), and the degree of increase of 15-F(2t)-IsoP in smoking group was significantly higher than that in non-smoking group(P<0.05). During reperfusion period (at T2,T3), the rate of lowering of 15-F(2t)-IsoP in smoking group was slower than that in non smoking group(P<0.05), and moreover, these patients needed more inotropic support than those in non-smoking group. CONCLUSION: Smoking patients have weaker anti oxidation capability during CABG, therefore they have higher incidence of low cardiac function. PMID- 15760531 TI - [Changes in content of inflammatory cytokines and troponin I in peri-operative periods of coronary artery bypass grafting with or without cardiopulmonary bypass]. AB - OBJECTIVE: To evaluate the effects of coronary artery bypass grafting (CABG) with or without cardiopulmonary bypass (CPB) on inflammatory response and myocardial injury. METHODS: Sixty patients undergoing elective CABG were randomly divided into CABG with CPB group (CCABG, 30 cases) and CABG without CPB group (OPCABG, 30 cases). Blood samples were collected from central vein to assay interleukin-6 (IL 6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), MB isoenzyme of creatine kinase (CK-MB) and troponin I (cTnI) before operation and 2, 8, 24, as well as 48 hours after operation, and all clinical data of the patients were recorded accordingly. RESULTS: After operation, the contents of IL-6, IL-8, TNF alpha, CK-MB and cTnI were significantly higher in CCABG group than those before operation and those in OPCABG group, while most of these parameters showed little change in OPCABG group both in operative or postoperative period. CONCLUSION: The results suggest that OPCABG may provide better myocardial protection and less systemic inflammatory response than CCABG. PMID- 15760532 TI - [Effect of combined decompression operation on middle and late stage cerebral herniation]. AB - OBJECTIVE: To observe the effect of combined decompression operation on patients with severe traumatic brain injury complicated by tentorial cerebral herniation. METHODS: Ninety-seven patients with an admission Glasgow Coma Scale score 3-5 were randomly divided into two groups: combined decompression group (n=46), in whom tentorium cerebelli was incised (2-4 cm) combined with bone flap craniectomy decompression [(10-15)cm x (15-17)cm], and conventional temporoparietal craniectomy group (n=51). CT scanning was performed in the patients before and after the operation. The patients of both groups received routine treatment and followed up for 1-32 months (mean 7 months) after the operation. The clinical symptoms, change in intracranial pressure and incidence of complications were compared between the two groups. RESULTS: The efficacious rate was 80.4% (37/46) in the combined decompression group, and among them 27 patients were cured (58.7%) and 10 patients remained to have moderate disability(21.7%). Nine patients (19.6%) died after combined decompression. However, in patients with conventional temporoparietal craniectomy decompression, favorable outcome was only found in 6 cases(11.8%), moderate disability accounted for 21.6% of patients, and 34 patients died(66.6%). In patients with combined decompression, the intracranial pressure was more efficiently lowered compared with conventional craniectomy decompression(P<0.01). Furthermore the incidence of acute brain edema, incisional herniation, traumatic epilepsy, occipital cerebral infarct and cerebro-spinal fluid(CSF) leakage were lower in combined decompression group compared with conventional craniectomy group (P<0.05 or P<0.01). The incidence of intracranial infection was not significantly different between two groups (P>0.05). CONCLUSION: Combined decompression is preferable to routine temporoparietal craniectomy for patients with severe head injury complicated by tentorial herniation. PMID- 15760533 TI - [Vascular endothelial growth factor expression in focal cerebral ischemia/reperfusion in injury in rats]. AB - OBJECTIVE: To study the vascular endothelial growth factor (VEGF) expression and its role in focal cerebral ischemia/reperfusion injury. METHODS: VEGF protein and mRNA expressions at the site of focal ischemia/reperfusion of rat cerebrum were assessed with immunohistochemistry and hybridization techniques. RESULTS: In normal cerebral tissue there was lower expression of VEGF. It was found to be expressed in the penumbra area after ischemia. With the prolongation of ischemia and ischemia/reperfusion, enhanced expression of VEGF tended to decline to normal level, while it enhanced in the penumbra area. CONCLUSION: Enhanced VEGF expression might play a protective role in focal ischemia/reperfusion injury to the cerebrum. PMID- 15760535 TI - [Change in humoral immunological function and their clinical significance in patients with cerebral hemorrhage]. AB - OBJECTIVE: To study the changes in humoral immunological function and their clinical significance in cerebral hemorrhage. METHODS: Among 115 cases of cerebral hemorrhage, there was low volume bleeding in 39 case, middle volume in 37 cases, and large volume in 39 cases. The ratios of CD19+-CD25+ and CD19+-CD25 B lymphocytes were determined by flow cytometry, and the content of IgA, IgG, IgM, and complement C3 were determined with scattered turbidimetry. The changes in humoral immunological function were compared in patients with different degrees of hemorrhage, and their variation in acute and recovery stages of disease were respectively assessed. RESULTS: The ratios of CD19+-CD25+and CD19+ CD25-B lymphocyte, and the levels of IgA, IgG, IgM, and C3 were higher than those of controls (all P<0.05). The ratios of CD19+-CD25+ and CD19+-CD25- B lymphocyte, and the contents of IgA, IgG,IgM and C3 were increased with the increase in amount of hemorrhage(all P<0.05). The ratios of CD19+-CD25+ and CD19+-CD25-B lymphocytes in recovery stage were lower than those in acute stage (all P<0.05). CONCLUSION: There are activations of humoral immunological functions in cerebral hemorrhage. The more abundant in hemorrhage and the severer in disease, the more predominant in the changes in humoral immunological functions. PMID- 15760537 TI - [Expression of caspase - 1 after hypoxic-ischemic brain damage]. AB - OBJECTIVE: To study the expression of caspase-1 mRNA in the cerebral cortex after hypoxic-ischemic brain damage(HIBD) in neonatal rats. METHODS: One hundred and twelve 7-day-old Wistar rats were randomly assigned to control group, HIBD 3 hours, 8 hours, 24 hours, 3 days, 6 days and 14 days groups (n=16 in each group), and standardized HIBD was given. In each group, 8 rats were used to assess the mRNA expression of caspase-1 in cerebral cortex by semi-quantitative reverse transcription-polymerase reaction, and another 8 rats were used to study histological changes with hematoxylin-eosin(HE) staining. RESULTS: The expression of caspase-1 mRNA was observed in the cerebral cortex in both control and HIBD groups. After HIBD, the level of caspase-1 mRNA in ischemic cortex began to increase at 24 hours (P<0.01 vs. controls), peaked at 6 days (P<0.01 vs. other groups) and decreased at 14 days. Histological study showed that the degenerated and necrotic neurons were increased progressively from 1 day to 6 days after HIBD, together with proliferation of glial cells. CONCLUSION: The increased expression of caspase-1 mRNA after HIBD, which was consistent with the time frame of the development of brain injury, indicates that it might play an important role in pathogenesis of HIBD in neonatal rats. PMID- 15760536 TI - [Influence of beginning time of hypothermia treatment on prognosis of extensive cerebral infarction]. AB - OBJECTIVE: To study the influence of beginning time of cranial hypothermia treatment on the prognosis of extensive cerebral infarction(ECI) so as to establish the optimum time for such treatment. METHODS: Ninety-two ECI patients were divided into three groups. In group A hypothermia treatment was begun within 6 hours after cerebral infarction in 31 patients. In another 31 cases in group B it was begun in 7-10 hours, and in 30 cases in group C it was begun in 11-14 hours after the attack. The mortality rate, the volume of cerebral infarction, neurological deficiency score(NDS) and quality of life in survivors were determined respectively in three groups. RESULTS: The volume of cerebral infarction in group A and B was obviously smaller than that of group C after hypothermia treatment(P<0.01). The mortality rate was higher in group C (26.67% in group C, 3.23% in group A and 6.45% in group B, both P<0.05) The mortality rate was highest in cases with high body temperature in group C(P<0.05). NDS was significantly lower in survivors of groups A and B compared with group C (both P<0.05), groups A and B compared with group C(P<0.05). The NDS and quality of life of the survivors with high body temperature(P<0.05 or P<0.01). CONCLUSION: Cranial hypothermia treatment should be begun with 10 hours after illness to obtain best effect. PMID- 15760544 TI - [Prevention and treatment of biliary complications in the perioperative period after liver transplantation]. PMID- 15760545 TI - [Evaluation of the changes of HBV serum markers and HBV DNA and the effects of perioperative factors after liver transplantation]. AB - OBJECTIVE: To study the changes of HBV markers and HBV DNA and the perioperative factors influencing them after orthotopic liver transplantation (OLT). METHODS: A retrospective study was undertaken. Data was collected from 97 patients in the First Affiliated Hospital of Sun Yat-sen University from March 1999 to October 2003. Patients were investigated on the 7-14, 14-30, 30-90, 90-180, 180-360 and 360- days after OLT. All the patients who received OLT were serum HBV positive before their operations. RESULTS: Kinetic expressions of HBV serum marker and HBV DNA were established. A few patient's HBeAg was negative (8%) before their operation. Within 7 day following surgery, no patient was HBeAg positive. However, the rate of HBeAg positive increased on the 90-180 day following surgery. The postoperation time of taking lamivudine was different between patients with HBeAg seroconversion and of those without (U = 88.5). Peaks occurred within 14 d of HBsAg negative and 14-30 d of anti-HBs positive after operation. Then they decreased and minimized at 90-180 day after liver transplantation. Patients who suffered more bleeding during the operation were more likely to be anti-HBs positive (3800ml vs. 3000ml, U = 8193.0) and HBsAg negative in serum within 2 week (5200ml vs. 4200ml, U = 1648.5) after OLT. While patient's who received more blood transfusion (1000ml vs. 1600ml, U = 9796.0) during operation were not likely to be anti-HBs positive in serum after surgery. Furthermore, the time of infusing HBIg did not affect the state of anti-HBs (U = 1252.5). At the same time, there were no correlations between the change of HBsAg in serum and in the method of operation (chi2 = 0.042). During this process, presentation of anti-HBc changed a little. CONCLUSION: The advantages brought on by operative factors become blunt 7-14 d following OLT. More attention should be taken to avoid reinfection of HBV 90-180 day after OLT. Tyrosine-methionine aspartic acid-aspartic acid (YMDD) mutation of HBV is more likely to occur when taking lamivudine longer. Then, HBV DNA should be monitored and a liver biopsy should be scheduled regularly after OLT. PMID- 15760546 TI - [Risk factors of renal failure in the early post-liver transplantation period]. AB - OBJECTIVE: To study the risk factors of renal failure in the early post-liver transplantation period. METHODS: 92 consecutive liver transplantation cases were reviewed and a multi-factor analysis of presumed risk factors of early post transplantation period renal failure was conducted. The factors analyzed were total bilirubin level, prothrombin activity, onset of structural renal disease, onset of gastrointestinal hemorrhage, whether the patient underwent large-volume paracentesis, or underwent plasmapheresis therapy, needed renal replacement therapy, the operation method used, the bleeding volume during operation and the immunosuppressive agents used. RESULTS: Of the 92 patients, 29 (31.5%) developed acute renal failure (ARF) in the early postoperative period. Multi-factor analysis revealed a high pre-transplantation serum creatinine level and low prothrombin activity as risk factors for development of ARF. CONCLUSION: ARF is a frequent medical complication after liver transplantation. A high pre transplantation serum creatinine level and low prothrombin activity are risk factors of its development. PMID- 15760547 TI - [Clinical analysis of aspergillosis in orthotopic liver transplant recipients]. AB - OBJECTIVE: To assess the clinical features of aspergillosis and its diagnosis, prophylaxis and treatment in patients after orthotopic liver transplantation (OLT), and to improve the prognosis of the recipients. METHODS: Medical records of consecutive patients who underwent OLT in our liver transplant center from May 2002 to May 2004 were analyzed retrospectively. Those with aspergillus infection complications were studied in detail regarding their infected organs, related factors, treatments and prognoses. RESULTS: 17 out of 207 recipients of OLT were detected with aspergillosis. The incidence was 8.21 percent. 5 patients infected with superficial aspergillus survived. Of the 12 cases with deep aspergillus infection, 3 with infection limited to the sites of their incisions survived, 2 of the 3 patients with infection in their lungs, and 1 of the 2 patients with it in their livers died, and 4 recipients with multi-organ aspergillus infection died. Among the 7 cases that died, 5 had severe hepatitis, 1 had post-hepatitis liver cirrhosis and 1 had primary liver carcinoma. CONCLUSIONS: Long-term (> or = 3 weeks) broad-spectrum antibiotics and immunosupression were involved in aspergillus infection in our OLT patients. Patients with chronic severe hepatitis had a higher risk of having aspergillus infection. Amphotericin B is still the best choice for treating aspergillosis. Prophylactic administration of anti fungal medicine, surveillance of fungal infections as a routine, and treatment of the infection in time may help to improve the prognosis of OLT recipients with aspergillosis. PMID- 15760548 TI - [Preoperation risk factor analysis in orthotopic liver transplantation with pre transplant artificial liver support therapy]. AB - OBJECTIVES: Orthotopic liver transplantation (OLT) is an accepted therapy for selected patients with advanced liver diseases. However, the early mortality rate after OLT remains relatively high due to the poor selection of candidates with various serious conditions. The aim of this study is to assess the value of pretransplantation artificial liver support treatment in reducing the pre operation risk factors relating to early mortality after OLT. METHODS: 50 adult patients in various stages of different etiologies who underwent OLT procedures had been treated with molecular adsorbent recycling system (MARS) preoperatively. The study was designed in two parts: the first one was to evaluate the effectiveness of a single MARS therapy by using some clinical and laboratory parameters which were supposed to be therapeutical pretransplantation risk factors. The second part was to study the patients undergoing OLT by using the regression analysis on preoperation risk factors relating to early (within 30 d after OLT) mortality rate. RESULTS: Among the 50 patients, a statistically significant improvement of the biochemical parameters was observed (pretreatment vs posttreatment). 8 patients cancelled their scheduled LTXs due to significant improvements in their clinical conditions or recovery of their failing liver functions. 8 patients died and 34 patients successfully underwent LTX. The immediate outcome (within 30 postoperative days) of these 34 patients was that 28 were kept alive and 6 died. CONCLUSIONS: Preoperation sequential organ failure assessment (SOFA), level of creatinine, INR, TNFalpha, and IL-10 are the main preoperative risk factors relating to early death after an operation. MARS treatment before a transplant operation can relieve these factors significantly, hence improve survival rate of liver transplantation or even make the transplantation unnecessary. PMID- 15760549 TI - [The effect of glycine on CD14 and NF-kappa B in Kupffer cells from rat liver grafts after ischemia-reperfusion injury]. AB - OBJECTIVE: To investigate the effect of glycine on CD14 and NF-kappa B in Kupffer cells from rat liver grafts after ischemia-reperfusion injury (IRI). METHODS: The rats were randomly divided into an IRI group, saline solution preconditioning group, and glycine preconditioning group. Their survival rates, graft functions, and hepatic histopathologic examinations were observed after IRI. Kupffer cells (KCs) following IRI were isolated and cultured to detect CD14 mRNA, NF-kappa B binding activity, and the TNF alpha and IL-1 level in the supernatant of the media. RESULTS: (1) Glycine preconditioning greatly enhanced the one-week survival rate (chi2 = 6.67 and 8.57 respectively), improved graft function, and ameliorated the histopathologic signs of injury. (2) The CD14 mRNA expression level (F = 7.64), NF-kappa B binding activity (F = 11.47), TNF alpha and IL-1 production (F = 14.08 and 9.56 respectively) in the glycine group were significantly lower than those in the other two groups. CONCLUSION: Glycine could efficiently protect rat liver grafts from ischemia-reperfusion injury by repressing the expression of CD14 and NF-kappa B binding activity in Kupffer cells and inhibiting the productions of TNF alpha and IL-1. PMID- 15760550 TI - [Construction of an adeno-associated virus vector expressing CTLA-4Ig and its expression in the transplanted liver allografts]. AB - OBJECTIVES: To construct a recombinant adeno-associated virus vector pSNAV expressing CTLA-4Ig and to demonstrate its expression in transplanted liver allografts and to see if a long term inhibitive effect of CTLA-4Ig could be obtained though its use. METHODS: After AAVCTLA-4Ig and PUC18 were cut with BamHI, CTLA-4Ig cDNA was inserted into the plasmid PUC18 by T4DNA ligase and PUC18-CTLA-4Ig was constructed. The obtained PUC18-CTLA-4Ig and pSNAV cut with Kpn I and EcoR I, CTLA-4Ig cDNA was inserted into plasmid pSNAV to construct the recombinant vector pSNAV-CTLA-4Ig, which was transfected into BHK-21 packaging cells by lipofectine-mediated transfection. Then the BHK-21 cell line was infected with HSV1-rc to produce a large amount of pSNAV- CTLA-4Ig. The specificity of the expressed product was identified by digestion with BamHI, PCR and sequence determination. The titer of the virus was detected. The product was infused into rats liver allografts via portal vein and its expression in the transplanted livers was detected immunohistochemically. RESULTS: Recombinant adeno-associated virus vector pSNAV-CTLA-4Ig was generated and purified into 8.5 x 10(11)/ml. Agarose gel analysis of PCR products verified the presence of CTLA 4Ig. Digestion with BamHI and sequence determination confirmed that pSNAV-CTLA 4Ig was constructed. Expression of CTLA-4Ig in the transplanted livers was detected successfully. CONCLUSION: Prepared pSNAV-CTLA-4Ig was constructed correctly and can express CTLA-4Ig effectively. Besides this, it can express CTLA 4Ig in rat liver allografts. It may be used in the study of transplant tolerance. PMID- 15760551 TI - [Binding and inhibition of adeno-associated virus Rep78 protein with hepatitis B virus C promoter]. AB - OBJECTIVES: Adeno-associated virus (AAV) Rep78 is known for its inhibitory effects on replication of several viruses and oncogenes transformations. The study was to investigate the effect of Rep78 on hepatitis B virus C (HBV-C) gene and the mechanism of it. METHODS: HBV-C promoter and HBV-C gene with its promoter were amplified by PCR and labeled with 32P-ATP. Electrophoretic mobility shift assay (EMSA) and in vitro transcription were utilized to detect the binding of MBP-Rep78 with HBV-C promoter and the transcription of HBV-C gene. RESULTS: EMSA showed that by increasing the amount of Rep78 protein from 0.1 microg to 1.0 microg, the binding bands got stronger in a dose-dependent manner. In addition, Rep78 antibody was used to certify the specificity of this binding. The compound of Rep78, Rep78 antibody and HBV-C promoter were seen as super shift bands in EMSA. Meanwhile, HBV-C gene transcription was significantly inhibited by in vitro transcription which meant that Rep78 could not only bind with HBV-C promoter, but also could inhibit the transcription of HBV-C gene. CONCLUSION: AAV Rep78 could inhibit the transcription of HBV-C gene through its binding with HBV-C promoter. PMID- 15760552 TI - [Relationship of CT manifestations of chronic viral hepatitis with severity of illness]. AB - OBJECTIVES: To study the relationship between CT imaging features and the severity of chronic viral hepatitis. METHODS: The clinical data and laboratory information of 120 chronic virus hepatitis patients were reviewed retrospectively. They were categorized into mild (34 cases), moderate (22), severe hepatitis (26) and hepatitis gravis (38) groups based on international standardized clinical criteria. All patients underwent standardized contrast enhanced spiral CT dual-phase scanning of the upper abdomen. The changes of the liver, bile duct, spleen, portal venous system, lymph nodes of the upper abdomen, peritoneal cavity and pleural cavity were examined and noted. Correlation analysis was applied for the relationship of CT imaging features and the severity indices of chronic virus hepatitis. RESULTS: As the severity of chronic hepatitis increased, which was reflected as serum albumin level went down and the value of ALT went up, the incidences of the following manifestations were also intensified: (1) intrahepatic perivascular lucency, (2) edema of the gallbladder fossa, (3) enlargement of abdominal lymph nodes, (4) ascites and (5) thoracic changes. Correlation between these CT manifestations and clinical severity of chronic viral hepatitis was of statistical significance. CONCLUSION: A close correlation exists between certain CT features of chronic hepatitis and the clinical severity of chronic viral hepatitis. PMID- 15760553 TI - [Investigation of the relationships between HBV YMDD variation and its genotypes]. PMID- 15760554 TI - [The prognostic value of cellular immunity function in patients with hepatocellular carcinoma]. AB - OBJECTIVES: To study the changes of cellular immunity function in patients with hepatocellular carcinoma (HCC) and its correlation with disease severity. METHODS: T lymphocyte subsets and CD28 co-stimulation molecule in CD8+ T cells in 22 HCC patients were detected using three-color flow cytometry. Serum interleukin 2 (IL-2), transforming growth factor beta 1 (TGFbeta1), and interleukin-6 (IL-6) were determined by ELISA and radioimmunoassay. A group of 30 patients with chronic hepatitis B (CHB), liver cirrhosis (LC), or normal adults (NC) served as controls. RESULTS: Compared with NC, the number of CD8+CD28- T cells increased and CD8+CD28+ T cells decreased in patients with HCC. The number of CD4+ T cells, CD4+/CD8+ ratios, IL-2 level all decreased and CD8+ T cells, IL-6, TGFbeta1 levels all increased in patients with HCC, LC and CHB. The CD4+ T cells, CD4+/CD8+ ratios and IL-2 level in patients with HCC were lower than those with CHB. Serum IL-6 and TGFbeta1 in patients with HCC were higher than those with LC and CHB. The levels of IL-6 and TGFbeta1 correlated with the stages of the tumors. CONCLUSIONS: HCC patients have a cellular immunity dysfunction. Rectifying the imbalanced function could be a potential way for treating HCC. Measurement of these factors would be useful for early diagnosis and evaluating the prognoses of these patients. PMID- 15760555 TI - [An in vitro study of specific antitumor immunity induced by dendritic cells pulsed with tumor cell lysates from patients with hepatocellular carcinoma]. AB - OBJECTIVE: To investigate T cell-mediated antitumor effects of dendritic cells (DCs) pulsed with tumor lysates in patients with hepatocellular carcinoma (HCC) in vitro. METHODS: DCs isolated from peripheral blood mononuclear cells of HCC patients were cultured and proliferated in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4), and then were pulsed with autologous hepatoma cell lysates. The phenotypes of the DCs were assayed by flow cytometry. The concentration of IFN gamma and IL-12 were measured in culture supernatants by ELISA. The ability of DCs pulsed with hepatoma cell lysates to stimulate proliferation of autologous T lymphocytes (CTL) was tested by thymidine incorporation method. The specific cytolytic activity of CTL was assessed by MTT method. RESULTS: The hepatoma cell lysates pulsed DC vaccines led to up-regulation of CD1a, CD40, CD86 and HLA-DR. Concentrations of IFN-gamma and IL-12 were increased more in the hepatoma cell lysate pulsed DCs group than those in the unpulsed DCs group, the hepatoma cell lysate group and control group. The proliferation of T-cells was markedly enhanced in the hepatoma cell lysate pulsed DCs group than that in the others. The CTL stimulated by the hepatoma cell lysate pulsed DCs had much higher cytotoxicity to autologous hepatoma cells (killing rate: 81.72%+/-9.49%), as compared with HepG2 and HNE-1 tumor cells (killing rate: 49.37%+/-11.21% and 17.14%+/-5.65%, respectively). CONCLUSION: The hepatoma cell lysate pulsed DC vaccines can induce an effective and specific anti-hepatoma effect. PMID- 15760556 TI - [Deletion of p15 and pl6 genes and overexpression of STK15 gene in primary hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the association of p15 and pl6 genes deletion and STKI5 gene overexpression in primary hepatocellular carcinoma (PHC). METHODS: The carcinoma tissue and the adjacent normal tissue were taken from 30 PHC patients during operations who had had neither chemotherapy nor radiotherapy preoperatively. DNA was extracted from the tissues and PCR was used to determine the homozygous deletion of p15 exon2 (pl5E2) and pl6 exon 2 (pl6E2). RNA was extracted, cDNA was synthesized by RT-PCR, and the expression of STKI5 gene was tested by PCR. Beta-actin was used as an internal control. Average density value (ADV) of STK15 gene and that of beta-actin gene were determined in both carcinoma tissue and the adjacent normal tissue. RESULTS: The rate of p15E2 deletion was 13.3% (4/30) and the rate of p16E2 deletion was 16.7% (5/30) in the carcinoma tissue. The p15E2 and pl6E2 co-deletion rate was 6.7% (2/30). In 19 of the 30 cases (63.3%) the expression of STK15 gene in carcinoma tissue was higher than that in the adjacent normal tissue. The ratio of ADV of STK15 gene to ADV of beta actin gene (1.53+/-0.31) in the carcinoma tissue was significantly higher than that (0.91+/-0.25) in the paired adjacent normal tissue (t = 2.86). CONCLUSION: The homozygous deletion of p15E2 and p16E2 and overexpression of STKI5 gene may play a role in the oncogenesis and malignant progression of PHC. PMID- 15760557 TI - [Expression of augmenter of liver regeneration in hepatic tumor cells and its clinical significance]. AB - OBJECTIVE: To investigate the effects of augmenter of liver regeneration (ALR) on the proliferation of hepatocytes and hepatic tumor cells and the expression of ALR in herpatocellular carcinoma (HCC). METHODS: Primary rat hepatocytes, QGY and HepG2 cells were cultured separately with ALR from different species. Cell proliferation was detected by their 3H-TdR uptake. The expression of ALR was examined in 9 normal hepatic tissues and 21 HCC cases using immunohistochemistry method. RESULTS: Different ALRs could stimulate the proliferation of HepG2 and QGY cells in a dose-dependent way in vitro, but all ALR had no influence in the proliferation of primary rat hepatocytes. The expression of ALR was absent in normal hepatic tissues, but present in all HCC hepatic tissues. However, the expression of ALR had no relationship with the differentiation and size of the carcinomas. CONCLUSION: ALR might play an important role in the occurrence and development of HCC. PMID- 15760558 TI - [Protective effect of SNMC on mice with fulminant liver failure]. AB - OBJECTIVE: To investigate the protective effect of stronger neo-minophagen C (SNMC) on fulminant liver failure (FLF). METHODS: D-Gal N and LPS were injected once into the abdominal cavity of rats to establish an experimental model of FLF. The level of plasma ALT, Alb, TBil, TNFalpha, NO, ET-1, IL-6 and liver histopathology of the rats were examined. RESULTS: In the D-Gal N and LPS model of FLF, there was an obvious decline of plasma TNFalpha (F = 52.84), NO (F = 15.81), ET-1 (F = 15.68), IL-6 (F = 15.32) and there was less hepatic tissue damage in SNMC-treated groups using different doses (high dose, medium dose, low dose) and at different times (pre-protection, simultaneous protection, post protection) compared with those not treated with SNMC. These results indicated that SNMC could be used to treat FLF. It was better to use a low dose of SNMC and use it at the same time as inducing the FLF. There were no differences in the results of those treated with SNMC of different dosages and treated at different times. CONCLUSION: SNMC can decrease the mortality of FLF by preventing hepatocyte apoptosis induced by D-Gal N and LPS and inhibit liver inflammation caused by all kinds of factors. PMID- 15760559 TI - [Isolation, cultivation and biological identification of human fetal marrow mesenchymal stem cells]. AB - OBJECTIVE: Noting the morphological and cytobiology characteristics and phenotypes of MMSCs, to establish an isolation and culture method for fetal MMSCs in order to provide a source of marrow mesenchymal stem cells (MMSCs). METHODS: Fetal MMSCs were isolated and cultured with in vitro cell culture technique; the characteristics of the proliferating and growing fetal MMSCs were studied with MTT and image analysis; the phenotypes of MMSCs were identified by flow cytometry and immunohistochemistry. RESULTS: Bone marrow of 12 fetuses was isolated within 0.5-2 h, and about 300+/-80 adherent cells were obtained at 24 h. Colonies with more than 5 cells were 15+/-6, growth detention period of culture cell was at 1-3 d after planting, log phase growth period was at day 4, and the amount of disintegration phase cells was reduced significantly. Original culture and serial subcultivations showed that cells divided prosperiously; unequal divisions special for stem cells were observed, and the amount of MMSCs harvested from each fetus was as much as 10(11)-10(12) cells after 10 serial subcultivations. The phenotype of MMSCs was CD166 positive and CD34 negative. Serial subcultivated MMSCs expressed a microamount of AFP and did not express albumine or CK18. CONCLUSION: Fetal MMSCs are easily isolated and proliferate prosperouly. Serial subcultivated MMSCs did not differentiate into hepatocyte-like cells under common culture condition and are feasibile as seed cells for tissue engineering reconstruction. PMID- 15760560 TI - [Comprehensive evaluation of the Chinese Journal of Hepatology (Zhonghua Ganzangbing Zazhi) from 1999 to 2003]. AB - OBJECTIVE: The Chinese Journal of Hepatology is a key journal in the research field of liver diseases in China. Ranked by the impact factor, which was issued and used by the Institute of Scientific and Technical Information of China, it is in fourth position among medical journals in China. In order to evaluate the journal, some facts about it were surveyed, including the number of pages, the number of papers in each issue, organizations of the authors, funding for their works, the impact factor, immediacy index, statuses of the articles' references, and a listing of their being cited. METHODS: The number of pages of each issue, the number of papers in every volume, and citations were quantitatively analyzed. Funding, impact factor, immediacy index, citations and organizations of the authors were analyzed by weighted Rank Sum Ratio (RSRw). RESULTS: In the five years, 1999, 2000, 2001, 2002, and 2003, (1) The Chinese authors came from 26 of the 31 provinces and cities in China. 48.8% in 1999 to 71.7% in 2003 of the authors were working in medical universities or medical colleges, and some authors were overseas experts. (2) The number of articles cited in the five years were 702, 1158, 1087, 1178 and 1744. (3) The number of papers published were 248, 221, 242, 212 and 336. (4) Impact factors of the journal were 0.897, 0.931, 1.421, 1.858, 1.440. With the cites, immediacy index, cited rate, ratios of research provided by national or international funds and number of organizations of authors evaluated, the RSRw results of the five years were 0.2750, 0.3417, 0.5000, 0.5000 and 0.5000. CONCLUSION: The Chinese Journal of Hepatology is well known and is one of the highest academic quality medical journals in China. It reflects the progress of liver disease research in China. PMID- 15760561 TI - [Problems and significance of liver allograft as an immunologically privileged organ]. PMID- 15760562 TI - [Selection of surgical methods used in liver transplantation]. PMID- 15760563 TI - [Acute cellular rejection in liver transplantation]. PMID- 15760564 TI - [Reinfection of HBV and its possible mechanism in patients with liver transplantation]. PMID- 15760565 TI - [Expressions of osteopontin and CD44v6 in hepatocellular carcinoma and their clinical significance]. PMID- 15760566 TI - [Study on a recombinant keratinocyte growth factor variant in treating experimental rat liver fibrosis]. PMID- 15760567 TI - [Influence of Fuzheng Huayu Capsules on abnormal amino acids spectrum in chronic liver diseases]. PMID- 15760568 TI - [The expression of GABA-A receptor subunit alpha1 beta1 gamma2 mRNA in hepatic encephalopathic rats]. PMID- 15760569 TI - [Effect of quercetin on the activities of cytochrome p450 1A1 in L-02 cell lines and human liver microsomes]. PMID- 15760570 TI - [Model for end-stage liver disease (MELD) score and liver transplantation]. PMID- 15760571 TI - [Plumbism following Chinese herbal treatment for chronic hepatitis B virus infection]. PMID- 15760572 TI - [Treatment of recurrent hepatitis C after liver transplantation]. PMID- 15760573 TI - [Hepatocarcinoma grew into the portal vein, inferior vena cava, right atrium, pericardium and peritoneum: a case report]. PMID- 15760574 TI - Gout: forgotten therapy for a common disease. PMID- 15760575 TI - Mechanisms of differential immunogenicity of tumor necrosis factor inhibitors. PMID- 15760576 TI - State-of-the-art disease-modifying osteoarthritis drugs. AB - Significant advances have occurred in the symptomatic management of osteoarthritis over the past several decades. However, the development of so called disease-modifying osteoarthritis drugs is in a more formative stage. Although increased knowledge of osteoarthritis pathophysiologic pathways provides more rational opportunity for targeting specific elements of the degenerative process, limitations in our ability to measure disease progression/regression hamper assessment. Development of more sophisticated plain radiographic techniques and the use of additional technologies such as magnetic resonance imaging and gadolinium-enhanced magnetic resonance imaging of cartilage provide potential for more reproducible approaches. Noninvasive biomarkers that reflect structural change are the subject of intense investigation. Studies describing disease-modification effects provide optimism that disease prevention, retardation, and reversal are attainable. PMID- 15760577 TI - How can one develop disease-modifying drugs in osteoarthritis? AB - A disease-modifying osteoarthritis drug should be able to interfere with the cartilage breakdown observed in this disease. The current question is whether development of a drug should be focused on structural parameters (eg, radiologic joint space width, magnetic resonance imaging cartilage volume, and so on), on clinical parameters such as functional impairment, or on a composite index such as the capacity of the study drug to prevent and/or to retard the requirement for total articular replacement. Another aspect of osteoarthritis trials is that several recent advances have greatly facilitated the conduct of phase I and II studies, such as biological markers of cartilage metabolism and magnetic resonance imaging parameters of cartilage breakdown. PMID- 15760578 TI - Considerations in the treatment of early osteoarthritis. AB - Osteoarthritis is the most common form of arthritis and it is one of the leading causes of disability in all elderly populations. It results in enormous societal burden, including the need for joint replacement at an annual cost to the community of billions of dollars. Two factors that predicate the formulation of a treatment strategy for an individual with "early" osteoarthritis include the risk for toxicity from long duration of exposure to pharmaceuticals and the desirability of a treatment with the potential to reduce the rate of disease progression. These considerations favor approaches that avoid or minimize chronic pharmaceutical use in favor of safer interventions, especially ones where current evidence suggests the potential of a disease-modification effect. These should include socio-behavioral interventions that promote weight optimization and exercise, consideration of orthotics, general dietary recommendations to increase vitamin D and C intake, and an emphasis towards topical or intermittent use of pharmaceutical agents rather than long term nonsteroidal anti-inflammatory drug use. PMID- 15760579 TI - Quality-of-life issues in osteoporosis. AB - Osteoporosis is responsible for more than 1.5 million fractures per year in the United States including vertebral and nonvertebral fractures of the hip and wrist. Most of the impact of osteoporosis on quality of life relates to fracture. While the morbidity and mortality of hip fractures have long been recognized, only recently have we also recognized the significant burden of vertebral fractures. With the development of generic and disease targeted quality-of-life instruments we have now confirmed observational studies showing that clinical vertebral fractures have significant effects on quality of life. The economic burden of vertebral fractures, if we include direct and indirect costs, may be similar to hip fractures. The relative risk for mortality after clinical vertebral fracture is at least equivalent to that of hip fracture. The burden of vertebral fractures relates not only to pain but also to kyphosis. Because vertebral fractures are often under diagnosed and under treated, these findings are a call to action because the first vertebral fracture leads to significantly increased risk for subsequent vertebral and osteoporotic fracture with further loss of quality of life. PMID- 15760580 TI - Premenopausal bone health assessment. AB - The World Health Organization criteria for classification of bone mineral density (BMD) cannot be applied to premenopausal women because the relationship between BMD and fracture risk is not the same as in postmenopausal women. Approximately 2.5% of premenopausal women have BMD that is more than 2.0 standard deviations below the mean BMD of an age-, gender-, and ethnicity-matched reference population. Most premenopausal women with low BMD have low peak bone mass and low 5- to 10-year probability of fracture. The management of these patients involves nonpharmacologic lifestyle measures and reassurances that fracture risk is low. A minority of premenopausal women with low BMD have significant elevation of fracture risk, usually a result of contributing diseases, conditions, or medications that may be identified and treated. Premenopausal women with fractures are at increased risk for postmenopausal osteoporosis and fractures later in life. PMID- 15760581 TI - Treatment of metabolic bone disease in patients with chronic renal disease: a perspective for rheumatologists. AB - As glomerular filtration rate (GFR) declines from age-related bone loss or disease that specifically induces a decline in GFR, there are a number of metabolic bone conditions that may accompany the decline in GFR. These metabolic bone conditions span a spectrum from mild-to-severe secondary hyperparathyroidism in early stages of chronic kidney disease (CKD) to the development of additional heterogeneous forms of bone diseases each with distinctly quantitative bone histomorphometric characteristics. Osteoporosis can also develop in patients with CKD and end-stage renal disease (ESRD) for many reasons beyond age-related bone loss and postmenopausal (PMO) bone loss. Diagnosing osteoporosis in patients with severe CKD or ESRD is not as easy to do as it is in patients with PMO. The diagnosis of osteoporosis in patients with CKD/ESRD must be done by first excluding other forms of renal osteodystrophy, through biochemical profiling or by double tetracycline-labeled bone biopsy and the finding of low trabecular bone volume. In such patients oral bisphosphonates seem to be safe and effective down to GFR levels of 15 mL/min. In patients with stage 5 CKD, who are fracturing because of osteoporosis or who are on chronic glucocorticoids, reducing the oral bisphosphonate dosage to half of its usual prescribed dosing for PMO seems reasonable from known bisphosphonate pharmacokinetics. However, we need better scientific data to fully understand bisphosphonate usage in this population. This paper deals with the evidence available to understand management of patients with CKD and opinions on what might be a reasonable clinical approach where evidence is currently lacking. PMID- 15760582 TI - Combination therapy for osteoporosis: considerations and controversy. AB - Combination therapy, the simultaneous use of two pharmaceutical agents with the goal being reduction of fracture risk, is an area of substantial clinical interest. This paper summarizes the rationale, existing clinical trials data, and other considerations relevant to combination therapy for osteoporosis. Combinations of antiresorbers (eg, estrogen plus bisphosphonates) produce greater increases in bone mass than either treatment alone. Conversely, combining anabolic agents (parathyroid hormone) with bisphosphonates does not produce additive effects. None of the existing studies are large enough to determine whether combination treatment reduces fracture risk to a greater extent than use of a single agent. However, it is certain that combination treatment will increase cost and likely that it will increase side effects and reduce therapy adherence. Given the absence of demonstrated fracture reduction benefit, increased cost and likely increase in adverse events, combination therapy is not currently recommended. PMID- 15760583 TI - Genetics of osteoporosis. AB - Genetic factors play an important role in regulating bone mineral density and other phenotypes relevant to the pathogenesis of osteoporosis such as ultrasound properties of bone, skeletal geometry, and bone turnover. Progress has been made in identifying quantitative traits for regulation of bone mineral density by linkage studies in man and mouse, but relatively few causal genes have been identified. Dramatic progress has been made in identifying the genes responsible for monogenic bone diseases and it appears that polymorphisms in many of these genes also play a role in regulating bone mineral density in the general population. Advances in knowledge about the genetic basis of osteoporosis and other bone diseases offer the prospect of developing new markers for assessment of fracture risk and the identification of novel molecular targets for the design of new drug treatments for osteoporosis. PMID- 15760584 TI - Oral antiresorptive therapy. AB - Oral antiresorptive agents play a pivotal role in the management of osteoporosis. This paper discusses the effects and potential future role of newer agents such as ibandronate. Alternative dosing schedules and routes of administration have become available and may improve fracture protection, compliance, and tolerability for the long term treatment of a chronic condition such as osteoporosis. Increasingly these agents are being used to reduce bone loss in other diseases associated with high risk for osteoporosis such as organ transplantation and cystic fibrosis. Such studies may act as prototypes for the extended use of this class of drugs in other chronic inflammatory disease states. The innovative, yet disappointing results from combining an antiresorptive agent (alendronate) with the anabolic effects of teriparatide is also discussed. The major problem that remains is the lack of direct comparison between the agents in terms of fracture endpoints. PMID- 15760585 TI - Osteoarthritis: a metabolic disease induced by local abnormal leptin activity? PMID- 15760586 TI - Outcomes and disease activity measures for assessing treatments in the idiopathic inflammatory myopathies. AB - The assessment and treatment of patients with idiopathic inflammatory myopathy has taken a major step forward over the past few years. Through the efforts of multi-disciplinary international groups of experts interested in the management of patients with myositis, initiatives have led to the development of a core set of outcome measures critical to their assessment. Similarly, the lack of consensus on several issues of clinical trial design has been addressed resulting in the development of a definition of clinical improvement for adult and juvenile patients with inflammatory myopathy using the core set outcomes. The final step in the puzzle of well-designed therapeutic trials in myositis is the determination of consensus guidelines to conduct such trials in adult and pediatric populations of myositis patients. PMID- 15760587 TI - Recent advances in juvenile dermatomyositis. AB - This paper reviews the knowledge we continue to gain in the areas of pathogenesis, evaluation of disease activity, and interventional strategies with regards to idiopathic inflammatory myositis. Potential new diagnostic criteria incorporate the use of magnetic resonance imaging and other imaging modalities along with the use of muscle biopsy pathologic features including that of immunologic markers. Specifically, this criterion reflects the upregulation of major histocompatibility complex molecules on muscle tissue during acute inflammation. PMID- 15760588 TI - Interstitial lung disease in polymyositis and dermatomyositis. AB - Interstitial lung disease occurs in approximately one-third of patients with polymyositis and dermatomyositis (PM/DM) and has an adverse effect on survival. It is commonly a component of early PM/DM and can precede the onset of muscle or skin disease. Its most common histopathology is nonspecific interstitial pneumonia. This is a more benign pattern, with respect to response to immunosuppression and also long-term survival, than the pattern of usual interstitial pneumonia seen in idiopathic pulmonary fibrosis. The clinical course of PM/DM lung disease is heterogeneous. Progressive and nonprogressive disease needs to be distinguished by clinical and physiologic monitoring to avoid over treatment. Patients with ongoing functional deterioration mostly benefit from immunosuppression. The experience with corticosteroid monotherapy is discouraging but cyclophosphamide, given as daily oral or intravenous pulse therapy together with corticosteroids, was found to be beneficial in many patients. Other immunosuppressants may be of benefit as well, but the weight of the current evidence supports the use of cyclophosphamide first. PMID- 15760589 TI - Imaging and skeletal muscle disease. AB - Imaging techniques have assumed increasing importance in the diagnostic approach to patients with muscle disease. These techniques include computed tomography, ultrasound, and magnetic resonance imaging (MRI). For most disorders of muscle, ultrasound and MRI are more useful than computed tomography. Advantages of ultrasound include accessibility at the bedside and lower cost. However, MRI remains the gold standard for detecting changes in muscle tissue. In some cases, MRI examinations can take the place of muscle biopsy for diagnosis. New advances in MRI include diffusion-weighted imaging, which permits assessment of fluid motion in muscles, and blood-oxygen-level-dependent imaging to evaluate tissue oxygenation. PMID- 15760590 TI - Exercise: an important component of treatment in the idiopathic inflammatory myopathies. AB - Resistive exercise is controversial for patients with idiopathic inflammatory myopathies. This paper provides an update on exercise and clinical assessment in these patients. The few published studies on this topic report unchanged disease activity from a variety of exercise regimens in patients in all stages of disease. Reduced disability was achieved in patients with polymyositis and dermatomyositis. In one study a slightly reduced impairment was reported in patients with inclusion body myositis, while in another study no objective reduction of disability could be detected. An increasing number of valid and reliable outcome measures are available for patients with polymyositis and dermatomyositis, but for patients with inclusion body myositis reliable and sensitive outcome measures are still needed. PMID- 15760591 TI - New hope for systemic sclerosis patients with digital ulcers. PMID- 15760592 TI - Genetics and proteomics in scleroderma. AB - The pathogenesis of systemic sclerosis (SSc; scleroderma) is still enigmatic. Genetic, familial, and twin studies suggest that SSc occurs in genetically susceptible individuals. Recent high-throughput technologies, including gene expression profiling and proteomics, have accelerated the rate of information acquired on possible mechanisms involved in SSc pathogenesis. The potential of the data obtained lies in their use for identifying new disease markers, monitoring disease activity, and developing tailored therapies. The result is an era of unprecedented advance that will benefit SSc and many other diseases. PMID- 15760593 TI - Clinical aspects of lung involvement: lessons from idiopathic pulmonary fibrosis and the scleroderma lung study. AB - Interstitial lung disease (ILD) occurs frequently among patients with systemic slcerosis (SSc) and accounts for significant morbidity and mortality. SSc-ILD resembles idiopathic pulmonary fibrosis (IPF) in many respects; each is characterized by a restrictive ventilatory defect that results from an active fibrosing process in the lung interstitium, and each is characterized by ground glass opacification on high resolution computed tomography (HRCT) and inflammatory cells in bronchoalveolar lavage fluid (BALF). Differences in the classification of lung histo-pathology are noted frequently and may account for some of the difference in outcome for patients with SSc-ILD and IPF. Optimal treatment of SSc-ILD remains to be determined, but cyclophosphamide has been reported to be effective in a number of case series. A randomized controlled trial, the Scleroderma Lung Study (SLS), will be completed in 2005; the outcome of the SLS should define the efficacy of daily oral cyclophosphamide for SSc-ILD patients with alveolitis defined by BALF and HRCT. PMID- 15760594 TI - Immunomodulatory therapy for SSc: will high-intensity immunosuppression with stem cell rescue improve outcome? AB - Effective treatments for severe systemic sclerosis (SSc) have not been identified. SSc is associated with high levels of morbidity and mortality related to functional impairment and organ dysfunction. The pathogenesis of SSc, as with all autoimmune disorders, is thought to involve the interplay between genetic background and environmental exposures. Abundant evidence exists for activation of humoral and cellular immunity in SSc. These immune and inflammatory mechanisms almost certainly play a role in vascular damage and activation of pathways that lead to fibrosis. However, the types and intensities of immunomodulatory therapies employed in other connective tissue diseases have not proved as useful in patients with SSc. The question remains if increasing the intensity of immunosuppression will prove a more successful strategy. Preliminary studies of intensification of therapy to immunoablative and myeloablative levels with hematopoietic stem cell rescue have been performed and appear promising. New studies to compare these approaches with conventional immunomodulatory therapy are underway. PMID- 15760595 TI - Animal models in scleroderma. AB - Scleroderma or systemic sclerosis is an insidious connective tissue disease with no known cure. A hallmark feature of scleroderma is the excess synthesis and deposition of collagen resulting in a fibrotic state. In scleroderma, fibrosis is not confined only to the skin but impacts internal organs as well. In an effort to better understand the pathophysiology of this disease, researchers have developed a variety of animal models that display features of the human condition. This paper focuses on mouse models of scleroderma and summarizes work conducted with these experimental paradigms that is focused on understanding the cellular and molecular events associated with the onset and maintenance of fibrosis. PMID- 15760596 TI - Scleroderma, fibroblasts, signaling, and excessive extracellular matrix. AB - Excessive extracellular matrix (ECM) deposition in the skin, lung, and other organs is a hallmark of systemic sclerosis (SSc). The pathogenesis of SSc is still poorly understood, but increasing evidence suggests that various cytokines such as transforming growth factor (TGF)-beta and their signaling pathways are key mediators of tissue fibrosis as a consequence of ECM accumulation in the pathogenesis of fibrosis such as SSc. TGF-beta regulates diverse biologic activities including cell growth, cell death or apoptosis, cell differentiation, and ECM synthesis. TGF-beta is known to induce the expression of ECM proteins in mesenchymal cells, and to stimulate the production of protease inhibitors that prevent enzymatic breakdown of the ECM. This paper focuses on the possible role of ECM, various cytokines, especially TGF-beta signal transduction pathways in the pathogenesis of fibrosis in SSc. PMID- 15760597 TI - [Portable coagulometers: a systematic review of the evidence on self-management of oral anticoagulant treatment]. AB - BACKGROUND AND OBJECTIVE: We aimed to systematically review the scientific evidence about the use of portable coagulometers for patient's self-management of oral anticoagulant treatment. MATERIAL AND METHOD: Systematic review of scientific evidence available from MEDLINE'S, DARE's, HTA-Database's, NHS-EED's and The Cochrane Library' s bibliographic databases, from their origin to March 2003. Randomized control trials (RCT) and Quasi-Experimental trials were selected provided that they compared patients in self-management with patients under usual care. The quality of scientific evidence was elicited using the Scottish Intercollegiate Guideline Network (SIGN) recommendations, whilst efficacy and security were descriptively summarized. RESULTS: Twelve (7 RCT and 5 quasi experimental trials) articles were found, and only two of them provided grade A recommendation. Patients under self-management remained the same or more time in the therapeutic range. The incidence of adverse effects in self-management patients was the same or less than that in patients under usual care. CONCLUSIONS: The quality of the scientific evidence is heterogeneous. For selected patients, patient's self-management is at least as effective and safe as usual care. New oral anticaogulants, which have shown promising results, should be scrutinized for future changes in service provision. PMID- 15760598 TI - [Evaluation of the pilot programme for the descentralization of control of oral anticoagulant treatment in the Osasunbidea Healthcare Service (Navarra, Spain)]. AB - BACKGROUND AND OBJECTIVE: The objective of this study was the evaluation of the pilot programme of decentralization of oral anticoagulant therapy (OAT) in eight basic health zones (ZBS) for the first six months and then a year after the programme was put into practice. PATIENTS AND METHOD: Descriptive transversal study. It includes all patients aged 14 years or older in the OAT (540 in the initial period and then 640 more) in eight ZBS (five urban and three rural). The evaluation was done including: prevalence of INR in control (2-3 or 2.5-3.5, according to indications), clinically suitable INR (INR in control +/- 0.2) and INR in control +/- 0.5, accumulated thrombosis and bleeding incidence. Source data: ANTICOAGN computer programme. RESULTS: Comparison of INR control between both periods: 59% against 63.9% of INR in the range of INR in control (p < 0.001), 72.8% against 78.8% for clinically suitable INR (p < 0.001), and 86.6% against 91.4% for INR in control +/- 0.5 (p < 0.001). Accumulated incidence of bleeding episodes in 6 months: Pilot: 1.1% (0.7% majors and 0.4% minors). Subsequent period: 3.6% (0.6% majors and 3% minors). CONCLUSIONS: The decentralization of control of the OAT with adequate resources implies a greater accessibility for the patient. The control of INR is acceptable and has improved significantly over the second period. We have detected an improvement in the increase of the INR below that of the range of the control INR. The incidence of minor hemorrhages has increased, owing probably to a better recording. PMID- 15760599 TI - [Effect derived by the adscription of internist to a surgical department]. AB - BACKGROUND AND OBJECTIVE: Although surgical units commonly request the collaboration of internists via medical consultation, the efficiency of this system is low and expensive. We studied the effect of the integration of full time internists in a surgical department. PATIENTS AND METHOD: The study group consisted of the patients admitted during intervention in the Orthopedic Surgery and Traumatology Department. Those patients admitted during the same period of the previous year made up the control group. We analyzed pre-surgical stay, post surgical stay and total stay. We also studied in-hospital mortality, re admissions and those patients who were not submitted to surgery (NSS). Control variables were age, sex, type of admission (programmed/emergency) and main diagnosis. RESULTS: 1,216 patients were included, 599 in the control group and 617 in the study group, 48.0% were emergency admissions and 11.7% NSS patients. Study of programmed patients did not suggest any differences between both groups in any of the analyzed variables. In emergency patients, the total stay was decreased in 18.2%, and it was reduced in 40.2% of the NSS. The distribution of the re-admissions was similar in both study and control groups. We observed a decrease in the NSS and a 50% decrease in the deaths of the study group. The obtained saving was 329,170 Euros. CONCLUSIONS: The adscription of full-time internists to a surgical service clearly improves the quality of the service it provides, with important hospital savings. PMID- 15760600 TI - [Factors influencing CRP levels in the diabetic population]. AB - BACKGROUND AND OBJECTIVE: The aim of the present study was to investigate the factors influencing CRP serum levels in the diabetic population. PATIENTS AND METHOD: Eighty-six patients with type 2 diabetes mellitus were prospectively included. The following variables were considered: age, gender, smoking habit, body mass index (BMI), glucose, HbA1c, total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), triglycerides, albumin excretion rate, CRP and E-selectin. RESULTS: CRP serum levels were associated with: gender (men: 1.4 mg/L [0.03-12.8] vs. women: 3.4 [0.07-80.7]; p = 0.02), age (r = 0.34, p = 0.001), BMI (r = 0.45, p < 0.001), triglycerides (r = 0.24, p = 0.03) and E-selectin (r = 0.21, p = 0.02). Those patients with at least one component of the metabolic syndrome, besides diabetes mellitus, had higher CRP concentrations in comparison with those patients without other components of the metabolic syndrome (2.4 mg/L [0.3-23.1] vs. 4.2 mg/L [0.6-80.7]; p = 0.05). In the multivariate analysis, gender (p = 0.001) and BMI (p = 0.01) were independently associated with the CRP concentration. CONCLUSIONS: Gender and BMI are the main factors to be considered when analyzing CRP levels in the diabetic population. Metabolic syndrome components other than diabetes are related to CRP levels in type 2 diabetic patients. PMID- 15760601 TI - [Oral therapy with anticoagulants: an endless history]. PMID- 15760602 TI - [Atypical antipsychotics: new etiology in type 2 diabetes mellitus]. PMID- 15760603 TI - [Is it appropriate to establish a recertification system for physicians in Spain?]. PMID- 15760604 TI - [Intravenous bisphosphonates]. AB - Bisphosphonates possess a fundamental role in the treatment of bone metabolic diseases. Yet their main limitations are poor oral absorption and gastrointestinal side effects, mainly esophageal irritation. Indeed, oral administration is unpleasant for many patients, and it is difficult in bed confined subjects. Therefore, intravenous administration of these agents can be very useful in several clinical scenarios, especially to improve the compliance. Recently, it has been showed that intravenous bisphosphonates are very useful to control hypercalcemia of malignancy, and to prevent bone complications related to metastases. Their use has also been analyzed in the prevention of bone disease after organ transplantation. Thus, their application to control Paget's disease of bone is well-known, and probably they could have an important role as antiresorptive agents in postmenopausal and steroidal osteoporosis. We present here a state of the art of the use of intravenous bisphosphonates for the aforementioned disorders. PMID- 15760605 TI - [Diminished antiepileptic drug serum levels during acyclovir treatment]. PMID- 15760606 TI - [Apolipoprotein in nonagenarian patients: association between alleles and genotype with prevalent diseases]. PMID- 15760607 TI - [Should HIV screening in pregnancy be repeated during the second and third trimester?]. PMID- 15760608 TI - [A patient with colorectal cancer and tuberculosis: false positive in PET and its clinical implication]. PMID- 15760609 TI - [Catemenial pneumotorax: recurrence after first dose of gonadotropin-releasing hormone (GnRH) analogues]. PMID- 15760610 TI - [Medical language and ethics]. PMID- 15760612 TI - The underdiagnosis of the vascular contribution to dementia. AB - The existence of vascular dementia (VaD) was first identified by Marie, who described the etat lacunaire, and by Binswanger, who identified white matter lesions in the brain subcortical areas. Alois Alzheimer, when defining the disease now bearing his name, did so in a patient with a presenile onset. The majority of demented elderly people were then believed to have cerebral arteriosclerosis underlying their cognitive decline. The role of cortical vascular lesions, while clear to clinicians, was highlighted only later, by the pathological studies of Tomlinson et al. and the clinical demonstrations of Hachinski et al. who have defined multi-infarct dementia. Lately, the emphasis shifted to pathogenic mechanisms for vascular brain disease with the identification of a multitude of processes, such as lipohyalinosis, cardiac dysfunction and genetic causes, to name only a few. Epidemiologic studies have demonstrated the high frequency of vascular lesions in brains of demented individuals, as well as the fact that vascular factors can contribute to Alzheimer's disease (AD). Moreover, many factors, which were identified as contributing to cerebrovascular disease in general and VaD in particular, are frequently suspected as predisposing to AD as well. All these considerations converge to the realization that vascular components are extremely important in the pathogenesis of old-age dementia and that prevention and perhaps treatment of dementia are within reach. These surprising findings highlight the importance of mixed vascular-degenerative dementia as a disorder that has to be properly defined. PMID- 15760613 TI - Cerebral microangiopathy in the mosaic of new discoveries. AB - After a short historical note on the diversion of the vascular concept of dementia from that of primary degeneration, this review describes cerebral deterioration on vascular grounds in three levels. Occlusions of big vessels with a single major infarct, middle calibre vasculopathy with multiple lacunes and finally microangiopathy causing leukoaraiosis--all these three entities produce cognitive disorders. Changes in the vessel wall, in cerebral blood flow velocity, the territorial and interterritorial rules for the location of malacia or lacunes are reviewed using neuroradiologic aspects revealed by ultrasound, by CT and MRI. Alterations in diffusion and their measurements by MRI are discussed in regard to ultrastructural reactions to ischaemia. Thresholds for tissue deterioration are shown in association with ATP depletion and with pathologic compounds in the interstitium. PMID- 15760614 TI - Cerebral small vessel disease: how does it progress? AB - Small vessel disease (SVD), or microangiopathy, of the cerebral white and central grey matter is an important subtype of vascular dementia (VD). SVD-dementia is characterised by a "dysexecutive" type of cognitive impairment, neurological deficits including imbalance and voiding dysfunction, and emotional disturbances. SVD is also frequent among clinically healthy subjects and patients with mild cognitive impairment. It is easily visualised by imaging techniques, but difficult to distinguish from mixed SVD/Alzheimer Disease. SVD has an inherent tendency to progress, but data on its natural course are sparse, and there are almost no drug trials dedicated to it. This article reviews the evidence on the speed and predictors of progression of SVD in regard to cognitive deficits, functional decline and white matter lesions, as derived from epidemiological, clinical and imaging studies and the placebo branches of VD drug trials. Based on the available data, we make suggestions for future research and outcome measures. PMID- 15760615 TI - Predictors of reversible mild cognitive impairment after stroke: a 2-year follow up study. AB - Many studies have investigated mild cognitive impairment (MCI) in the context of prodromal dementia, but few have investigated recovery from MCI. The aim of this study was to determine the prevalence of reversible MCI after stroke and to identify factors related to recovery. METHODS: One hundred and eighteen patients with a first ever cerebral stroke were followed up for 2 years. Neuropsychological assessment was performed at 1, 6, 12, and 24 months poststroke. Possible predictors of reversible MCI were demographic variables, baseline MMSE scores, presence of stroke risk factors, and CT variables. Poststroke MCI was diagnosed when there was a deficit in at least one cognitive domain, without their being demented. Recovery was considered when MCI was no longer present. RESULTS: Twenty-four (20.3%) patients were classified as having permanent reversible MCI and were compared with patients without recovery. Most patients recovered from MCI between the first and second assessments (19.7% versus 13.1% and 2.0% later on). Higher baseline MMSE scores and female sex were independent predictors of recovery (OR(High MMSE)=9.9; OR(female sex)=2.8). Neither stroke-related risk factors nor CT variables were predictors of favorable outcome. CONCLUSION: About 20% of patients with poststroke MCI recover from MCI. Higher MMSE scores at baseline and female sex are independent predictors of this recovery. PMID- 15760616 TI - Noninfarct vascular dementia and Alzheimer dementia spectrum. AB - Vascular dementia is an overarching superordinate category of which multiinfarct vascular dementia is only one subtype. To contribute to the definition of vascular dementia, method involved investigation of mental status, oral language and comprehension in 81 consecutive vascular patients comprising two vascular samples: cerebral infarct sample (n=43) and cerebral noninfarct sample (n=38). To determine baseline, method also involved investigation of 36 demographically equivalent normal elderly. Results indicate both vascular samples performed significantly worse than normal elderly. Results further indicate there were no robust, reliable, significant differences between cerebral infarct and cerebral noninfarct patients. The lack of significant differences between cerebral infarct and cerebral noninfarct vascular samples brings into focus the ambiguous transition between diffuse, generalized disease and the multifocality underlying the vascular dementia-Alzheimer dementia spectrum. Cross-cutting infarct and noninfarct vascular populations were vascular factors of arteriosclerosis, abnormal blood pressure, diabetes mellitus, abnormal electrocardiogram, peripheral vascular disease, and other variables implicated in the distal causality of both infarct and noninfarct vascular dementias. Results indicate cerebral infarction is not the only path to the final common phenotype of vascular dementia. Vascular dementia is reconceptualized so as to include noninfarct vascular dementia: vascular dementia caused by underlying vascular factors other than cerebral infarction. It is suggested that one form of the subtype of noninfarct vascular dementia is Alzheimer-type vascular dementia. PMID- 15760617 TI - Prevalence and pathogenic role of cerebrovascular lesions in Alzheimer disease. AB - Cerebrovascular lesions in Alzheimer disease (AD) being significantly more frequent than in nondemented elderly subjects suggest overlaps and synergistic effects between both pathologies. Examination of a consecutive series of autopsy proven AD cases and age-matched controls revealed a higher frequency of vascular lesions and of cerebral amyloid angiopathy (CAA) in AD (57.34% vs. 33.2% and 94.1% vs. 33.3%, respectively). These and previous data on vascular pathology in Parkinson disease emphasize its importance in these disorders. A study comparing the frequency and extent of general CAA and capillary CAA (CapCAA) in the postmortem frontal cortex of cases with high and low Braak stages showed no correlation between general CAA and dementia, only a low one with other vascular lesions except for cerebral hemorrhages. However, it was higher in AD than in controls with vascular pathology. The severity of CapCAA not correlating with general CAA showed high correlation with AD pathology, suggesting different pathogenesis of both types of CAA. Its elucidation may have implications for new therapeutic strategies. Considering the variability of vascular pathology in both AD and aged brains, the mechanisms behind their interactions are largely unknown, and further studies are needed to clarify their impact on cognitive impairment. PMID- 15760618 TI - Survival following dementia onset: Alzheimer's disease and vascular dementia. AB - Survival following the onset of dementia has been reported to vary from 3 to over 9 years. We examined mortality in 3602 participants of the Cardiovascular Health (CHS) Cognition Study in four US communities evaluated for dementia incidence between 1992 and 1999 and followed for 6.5 years. By June 2000, 33 of 62 (53.2%) participants who developed vascular dementia (VaD) had died compared to 79 of 245 (32.2%) with Alzheimer's disease (AD), 66 of 151 (43.7%) with both AD and VaD, and 429 of 2318 (18.5%) with normal cognition. Using Cox proportional hazards regression with a time-dependent covariate for dementia status adjusted for age, gender and race, individuals with VaD were more than four times as likely to die during follow-up than those with normal cognition (HR: 4.4, 95% CI: 3.1-6.3). The hazard ratios were 2.1 (95% CI: 1.6-2.7) for AD and 2.5 (95% CI: 1.9-3.3) for both types. Adjusted accelerated life models estimated median survival from dementia onset to death as 3.9 years for those with VaD, 7.1 years for AD, 5.4 years for mixed dementia, and 11.0 years for matched controls with normal cognition. While persons with VaD died primarily from cerebrovascular disease, those with AD/mixed dementia died more frequently from dementia/failure to thrive. PMID- 15760619 TI - The acoustic cortex in vascular dementia: a Golgi and electron microscope study. AB - Morphological alterations in vascular dementia have been extensively described in the hippocampus, the cerebral cortex, the subcortical centers and the cerebellum. In the present study, we describe the morphological alterations of the acoustic cortex in five cases, which fulfilled the clinical, neuropsychological and laboratory diagnostic criteria of vascular dementia. The morphological alterations, seen in Golgi technique and electron microscopy concerned the capillaries, the dendritic arborization of the neurons, the astrocytes and the cytoarchitecture of the cortex. The neurons showed an impressive abbreviation of the dendritic fields and loss of spines. Astrocytic proliferation was seen in the cortex. The layer I showed marked decline of Cajal-Retzius cells. The majority of the synapses demonstrated changes in size and shape of the pre- and postsynaptic components and alterations of the organelles. The morphological alterations of the acoustic cortex in vascular dementia may be associated with the impairment of the verbal communication, which is not an uncommon phenomenon even in the early stages of the vascular dementia. PMID- 15760620 TI - Understanding the pathology of vascular cognitive impairment. AB - The prevalence, morphology and pathogenesis of vascular dementia (VaD), recently termed vascular cognitive impairment (VCI), and of mixed dementia (Alzheimer disease+vascular encephalopathy) are a matter of discussion and no validated neuropathologic criteria for these disorders are currently available. In Western memory clinic-based series, VaD/CVI is suggested in 8-10% of cognitively impaired elderly; its prevalence in autopsy series ranges from 0.03% to 58% (mean 5-15%). Fairly unusual as an isolated nosological entity, CVI appears to correlate with focal, multifocal or diffuse cortical and/or subcortical microinfarcts and lacunes often affecting strategically important brain areas (thalamus, frontobasal, limbic system), hemispheric white matter and, less often, large brain areas. They result from systemic, cardiac or local large or small vessel disease. The lesion pattern in "pure" VCI with predominant multiple small (subcortical) lesions related to microangiopathies differs from that in "mixed dementia" (AD+VaD), more often associated with large infarcts, suggesting different pathogenesis. In very old subjects, selective hippocampal sclerosis may be accompanied by multiple other vascular pathologies. Minor cerebrovascular lesions (CVL), except for severe amyloid angiopathy, appear not essential for cognitive decline in full-blown AD, while both mild AD-type pathology and small vessel disease may interact synergistically in "unmasking" or promoting dementia. AD pathology is significantly less severe in the presence of cerebrovascular lesions. Further studies are needed to validate diagnostic criteria for VCI and to clarify the impact of vascular lesions on cognitive impairment. PMID- 15760621 TI - Coronary artery bypass grafting (CABG) and cognitive decline: a review. AB - Coronary artery bypass grafting (CABG) is a worldwide used myocardial revascularization procedure, which despite the modern advantages still has a spectrum of complications. The relatively old age of the patients who undergo CABG and their widespread atherosclerotic disease are possible reasons for vascular sequels particularly those leading to neurological dysfunction. Immediate, early and late cognitive decline is a known- and well-defined outcome of CABG. Whereas a lot of data was collected through the last years regarding to neuropsychological dysfunction after CABG, nevertheless etiology and mechanisms of this phenomenon remain unresolved and they are probably multifactorial. Meticulously preoperative assessment of those patients with a potential risk of adverse neurocognitive outcomes can help clinicians to select the mode of revascularization and to better counsel patients about the risks and benefits of surgery versus more conservative kinds of treatment. PMID- 15760622 TI - Methods of detecting potential causes of vascular cognitive impairment after coronary artery bypass grafting. AB - Coronary artery bypass grafting (CABG) is the most common major surgical procedure performed worldwide. Neuropsychological deficits are frequent following CABG occurring in up to 80% in the early postoperative period, 20-50% at 6 weeks and 10-30% of patients at 6 months. Transcranial Doppler monitoring is well suited for monitoring the brain during surgery. It has been shown that both solid and gaseous microemboli are frequent during, surgery especially during clamping and declamping of the aorta. This method can also monitor cerebral hemodynamics during surgery and alert the surgical team when a fall in perfusion pressure occurs. Magnetic resonance imaging (MRI) studies have found evidence which suggests increased water content in the brain following "on-pump" CABG. New postoperative cerebral lesions have also been found in many patients using diffusion-weighted MRI. PMID- 15760623 TI - Patterns of memory impairment and perseverative behavior discriminate early Alzheimer's disease from subcortical vascular dementia. AB - Previous research suggests that the neuropsychological deficits in Alzheimer's disease (AD) are different from that of vascular dementia (VaD), especially with respect to memory, language and executive functions, but negative findings were reported. Our objective was to clarify the cognitive syndrome in AD and VaD in the early stage of these disorders. We investigated 45 patients with early AD, 23 patients with subcortical VaD and 35 normal controls. All subjects were assessed with neuropsychological battery designed to measure memory, language, praxis and executive functions. Patients with AD had significantly worse scores on Story Recall (p<0.02) and on all measures of the Free and Cued Selective Reminding Test (p<0.03 to 0.001) than did patients with VaD, as well as greater number of perseverations (p<0.02) on category fluency. Conversely, VaD patients had more perseverations (p<0.02) on the Modified Card Sorting Test. Despite the similar degree of overall cognitive deterioration, the findings show more impaired retrieval from long-term storage in AD than in VaD. Moreover, the data suggest that AD and subcortical VaD affect perseverative behavior in a different fashion. These results may be helpful in differentiating AD from VaD in the early stage of these disorders, when mental impairments are not pervasive yet. PMID- 15760624 TI - From trepidant abasia to motor network failure--gait disorders as a consequence of subcortical vascular encephalopathy (SVE): review of historical and contemporary concepts. AB - Gait disorders in progressive subcortical vascular encephalopathy (SVE) and their impact on the burden of disability in the growing elderly population are underrepresented in medical scientific literature. The absence of a clear framework for the diagnosis and classification for gait disorders on the basis of SVE has multiple reasons: (1) neither movement disorder specialists nor stroke specialists are truly familiar with this topic and feel responsible for its treatment, (2) the existing terminology lacks a clear concept and a consistent classification, and (3) only in recent years have large prospective trials started to address the natural course of SVE. This article reviews the classical descriptions of gait disturbances with preferential view to our present concept of SVE, and comments on historical and current nosology of gait disorders aiming to propose for a new classification. PMID- 15760625 TI - Gait slowing as a predictor of incident dementia: 6-year longitudinal data from the Sydney Older Persons Study. AB - Current definitions for the preclinical phase of dementia focus predominantly on cognitive measures, with particular emphasis on memory and the prediction of Alzheimer's disease. Incorporation of non-cognitive, clinical markers into preclinical definitions may improve their predictive power. The Sydney Older Persons Study examined 6-year outcomes of 630 community-dwelling participants aged 75 or over at recruitment. At baseline, participants were defined as demented, cognitively intact or having a syndrome possibly representing the preclinical phase of Alzheimer's disease, vascular dementia, an extrapyramidal dementia or various combinations of the three. Those with cognitive impairment in combination with gait and motor slowing were the most likely to dement over the 6 year period (OR 5.6; 95% CI 2.5-12.6). This group was also the most likely to die (OR 3.3; 95% CI 1.6-6.9). White matter indices on MRI scanning were not consistently correlated with gait abnormalities. Simple measures of gait may provide useful clinical tools, assisting in the prediction of dementia. However, the underlying nature of these deficits is not yet known. PMID- 15760626 TI - Optimized neuropsychological procedures at different stages of dementia diagnostics. AB - OBJECTIVE: To establish a staged procedure in dementia diagnostics and to propose specific, abbreviated test batteries suitable for the three diagnostic stages: Primary medical care, neuro-psychiatry, and memory clinic. METHODS: A total of 159 participants underwent comprehensive clinical, neurological, neuropsychological, and MRI examinations. The neuropsychological examination took approximately 90 min per individual and was based on tests of verbal and visual memory, language, abstract thinking, attention, visuo-constructive and spatial functions. Stepwise discriminant analyses were performed to identify which subset of the 18 variables of the comprehensive test battery was the most appropriate to differentiate between specific diagnostic groups, and which variables could be discarded to abbreviate the test battery without substantial loss in diagnostic accuracy. RESULTS: The abbreviated versions of the test battery retained adequate diagnostic accuracy. The accuracy decreased by maximally 4%, whereas the test administration time dropped substantially from previously 90 min to a maximum of 50 min. CONCLUSION: Depending on the diagnostic question, a specifically abbreviated version of the comprehensive test battery can be used without unacceptably reducing diagnostic accuracy. PMID- 15760627 TI - Differential cognitive outcomes in the Hypertensive Old People in Edinburgh study. AB - Hypertension is associated with cognitive impairment in older adults. The Hypertensive Old People in Edinburgh (HOPE) study reported improved scores in two psychometric tests in those subjects with the greatest fall in diastolic blood pressure during a 24-week randomised, double-blind trial of captopril versus bendrofluazide in 81 elderly hypertensive people with mild cognitive impairment. Three hundred and eighty-seven of the original sample of 603 older people with and without hypertension and/or cognitive impairment from which the trial subsample was drawn were available for adequate psychometric testing 4 years later. Blood pressure was related prospectively to Raven's Progressive Matrices (RPM), a measure of fluid intelligence, but not memory differences. RPM scores were obtained at baseline, 6 weeks, 12 weeks and at the end of the randomised controlled trial. For subjects on captopril mean scores at each time point adjusted for blood pressure change were 27.6 (95% CI 25.5-29.6), 27.2 (95% CI 25.1-29.2), 28.4 (95% CI 26.6-30.3) and 28.9 (95% CI 26.9-30.9), and for bendrofluazide 27.1 (95% CI 25.1-29.0), 28.9 (95% CI 26.9-30.9), 28.9 (95% CI 27.2-30.7) and 28.7 (95% CI 26.8-30.6). There was a significant improvement in scores for those on bendrofluazide compared with captopril at week 6 (F=8.10, p=0.006, partial eta2=0.11). There were no significant effects for either drug or blood pressure at any time point for tests of memory. Future trials of the effects of antihypertensive therapy on cognition should focus more on outcomes other than memory. Early differential effects of therapeutic agents may not be maintained. PMID- 15760628 TI - Effects of arousing emotional scenes on the distribution of visuospatial attention: changes with aging and early subcortical vascular dementia. AB - BACKGROUND: The modulation of attention by emotionally arousing stimuli is highly important for each individual's social function. Disturbances of emotional processing are a supportive feature for the diagnosis of subcortical vascular dementia (SVD). We address here whether these disturbances might be useful as an early disease marker. METHODS: In order to examine the modulation of visual attention by emotionally arousing stimuli of different valence, 12 elderly patients with early SVD, 12 age-comparable healthy adults and 12 young healthy subjects were studied while looking at pairs of pictures from the International Affective Picture Battery that were either neutral-neutral, neutral-positive or neutral-negative in terms of emotional content. Eye movements were recorded with an infrared eye-tracking system. The direction of the first saccade and the dwell time during the 10 s of presentation were measured and compared among groups with parametric tests. RESULTS: All subjects showed a modulation of initial attentional orienting as well as a higher percentage of dwell time towards the pictures containing emotional material. Patients with SVD and old controls did not differ in either experimental measure. Young patients showed a stronger bias towards emotionally negative material than both groups of older individuals. CONCLUSIONS: Modulation of visuospatial attention is preserved in early SVD. This might have implications for therapeutic interventional approaches. A weakened sustained attention towards negative but not positive emotional pictures in the elderly is in accordance with the socioemotional selectivity theory, describing a relative selection of positive stimuli with aging. PMID- 15760629 TI - Dysexecutive syndrome following anterior thalamic ischemia in the dominant hemisphere. AB - In the article, we describe a rare case: a 52-year-old male patient with an isolated lesion of the anterior thalamic nuclei in the dominant hemisphere due to an ischemic stroke. The patient's syndrome of disordered verbal fluency, working memory, learning and executive functions is seen as a part of a lesion in the dorsolateral complex circuit. During the following 6 months, we observed significant improvement in visual learning, recent memory and categorisation of subjects, while verbal learning and visuospatial planning remained insufficient. In this context, the preservation of thalamic structures in different types of cognitive deterioration is discussed. PMID- 15760630 TI - MRI identifies MCI subtypes: vascular versus neurodegenerative. AB - As life expectancy increases worldwide, pandemics of cognitive impairment and dementia are emerging as major public health problems. Mild cognitive impairment (MCI), prodromal for dementia, is a descriptive term used for those clinical states showing early and subtle cognitive decline among the elderly, preceding the dementias. Psychometric screening combining Mini-Mental Status (MMSE) and Cognitive Capacity Screening (CCSE) Examinations, when combined now called CMC with C standing for Combined, M for MMSE, and C for CCSE, confirms diagnosis of MCI. Individuals identified with MCI are at increased risk for dementia of Alzheimer's type (DAT), vascular dementia (VAD) and other rare neurodegenerative dementias, including Lewy body dementia (LBD), fronto-temporal dementia (FTD) and Parkinson's disease dementia (PDD). Depending on different clinical compositions of cohorts studied, and MCI criteria used, between 19% and 50% of MCI progress to dementia within 3-5 years, two thirds to Alzheimer's (DAT) and one third to vascular types (VAD) in the United States. Not all Parkinsonians become demented, but PDD develops in 20-30%, however, PDD subjects were excluded from the present study, as were LBD and FTD. The incidence of MCI in those over age 70, with confirmed cognitive declines when tested 2 years later, is around 23%, but some spontaneously improve. PMID- 15760631 TI - Magnetic resonance spectroscopy in patients with MELAS. AB - Localized magnetic resonance spectroscopy (MRS) yields sensitive metabolic markers to provide insight into the pathophysiology of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) in vivo. Findings in full MELAS syndrome at 1H MRS of the brain typically include severely elevated lactate and reduced N-acetylaspartate, glutamate, myo-inositol, and total creatine concentrations in stroke-like lesions. Similar but less extreme alterations are also common in gray matter (GM) regions that appear normal at magnetic resonance imaging. Phosphorus spectroscopy of peripheral muscle permits investigation of the bioenergetic status. A decline of the phosphorylation potential indicates a low energy reserve at rest. Phosphocreatine resynthesis during post-exercise recovery is delayed pointing to reduced mitochondrial capacity. As MRS is inherently non-invasive, follow-up studies can be performed to assess treatment response quantitatively. PMID- 15760632 TI - Cognitive, global, and functional benefits of donepezil in Alzheimer's disease and vascular dementia: results from large-scale clinical trials. AB - Alzheimer's disease (AD) and vascular dementia (VaD) are both associated with deficits in cholinergic neurotransmission that are amenable to therapeutic intervention. The cholinesterase inhibitor, donepezil, is clinically effective in both AD and VaD. Results from a 10-study metaanalysis of donepezil (5 or 10 mg/day) in AD and a two-study combined analysis of donepezil (5 or 10 mg/day) in VaD are presented to compare patient characteristics and donepezil treatment outcomes. The analyzed studies were randomized, placebo-controlled, and of up to 24 weeks duration. In both AD and VaD, donepezil provided significant benefits compared with placebo on measures of cognition and global function. Placebo treated AD patients showed a decline in cognition and global function, whereas placebo-treated VaD patients remained stable, suggesting treatment effects of donepezil in VaD were driven by improvement rather than stabilization or reduced decline. More VaD patients than AD patients received concomitant medications. Cardiovascular adverse events were more common in VaD than AD patients but were not increased by donepezil. In conclusion, although there are differences between AD and VaD patients in comorbid conditions and concomitant medications, donepezil is effective and well tolerated in both types of dementia. PMID- 15760633 TI - A study of statin use in the prevention of cognitive impairment of vascular origin in the UK. AB - INTRODUCTION: There is increasing evidence suggesting that control of hypertension and antiplatelet therapy may prevent or reduce progression of vascular-related cognitive impairment (VCI), though a similar role of statins in this group of patients has yet to be established. METHODS: We conducted a postal survey of a group of 296 medical practitioners (comprising of physicians and psychiatrists specialising in the elderly, and general practitioners) inquiring into their management of (a) patients at high risk of developing VCI and (b) patients with established VCI. RESULTS: The overall response rate was 60% (177/296), with the highest response rate from psychiatrists. (a) For patients at high risk of developing VCI: 47% of clinicians believed that statins had an important role in preventing subsequent dementia: 4% would commence statins at a total cholesterol (TC) of 4-5 mmol/L; 38% with a TC of 5.1-6.5 mmol/L; and 32% with a TC of 6.6-8 mmol/L. Cardiovascular risk profile, age, cost and gender were other factors considered as important factors influencing statin prescription. (b) In those patients with established VCI: 32% of clinicians felt that statins had an important role in arresting progression, usually in people with a mild degree of cognitive impairment: 4% would commence statins at a TC of 4-5 mmol/L; 25% with a TC of 5.1-6.5 mmol/L; and 22% with a TC of 6.6-8 mmol/L. There were no major differences between clinicians in their prescribing habits. CONCLUSIONS: A substantial proportion of clinicians favour the use of statins in primary and secondary prevention of cognitive impairment of vascular origin, despite a lack of definite evidence to support their use at the present time. PMID- 15760634 TI - Does blood pressure lowering treatment prevents dementia or cognitive decline in patients with cardiovascular and cerebrovascular disease? AB - There is increasing evidence that both hypertension and stroke play important roles in the development of cognitive decline and dementia. Despite five high quality randomised controlled trials (RCTs) in this area to date, there remains uncertainty about the role of blood pressure lowering therapy in the prevention of cognitive decline and dementia. It appears that lack of definitive results from these trials can be explained on the basis of (a) insufficient power to detect modest treatment effects; (b) measurement error in the diagnosis of dementia; (c) variations in the treatment effects between different types of antihypertensive agents; and (d) bias due to missing data, variation in baseline factors such as levels of blood pressure, and the inclusion of patients with cognitive impairment at entry. Preliminary meta-analysis of RCTs supports the hypothesis that blood pressure lowering may prevent dementia in high-risk patients, that is those with vascular disease. However, a meta-analysis of individual patient data (IPD) from these, and other relevant trials in patients with vascular disease, would provide much more reliable data. If the hypothesis were confirmed, it would certainly be of considerable importance not only in terms of our understanding of the aetiology of dementia, but also in promoting blood pressure lowering strategies for broader public health good. PMID- 15760635 TI - Cognitive functioning after repetitive transcranial magnetic stimulation in patients with cerebrovascular disease without dementia: a pilot study of seven patients. AB - AIMS: Examine whether one session of high frequency repetitive transcranial magnetic stimulation (rTMS) applied over the left dorsolateral prefrontal cortex (DLPFC) would induce any measurable cognitive changes in patients with cerebrovascular disease and mild cognitive deficits. PATIENTS AND METHOD: Seven patients with cerebrovascular disease and mild executive dysfunction entered the randomised, controlled, blinded study with a crossover design. rTMS was applied either over the left DLPFC (an active stimulation site) or over the left motor cortex (MC; a control stimulation site) in one session. Each patient participated in both stimulation sessions (days 1 and 4) and the order of stimulation sites (DLPFC or MC) was randomised. A short battery of neuropsychological tests was performed by a blinded psychologist prior to and after each rTMS session. Psychomotor speed, executive function, and memory were evaluated. RESULTS: The only mild but significant stimulation site-specific effect of rTMS was observed in the Stroop interference results (i.e. improvement) after the stimulation of DLPFC but not MC in comparison with the baseline scores (Wilcoxon, Z=-2.03, p=0.04). Patients improved in the digit symbols subtest of the Wechsler adult intelligence scale-revised after both rTMS sessions regardless of the stimulation site (DLPFC or MC; Z=-2.06, p=0.04 and Z=-2.06, p=0.04, respectively). There was no measurable effect of rTMS in any other neuropsychological test. CONCLUSION: Our pilot study results showed that one session of the high frequency rTMS applied over the left DLPFC was safe in patients with cerebrovascular disease and mild executive deficits, and may induce measurable positive effects on executive functioning. PMID- 15760636 TI - Aspirin resistance in stroke: 2004. AB - Aspirin is a well-established medication in the treatment of atherothrombotic vascular disease. However, despite aspirin treatment a substantial number of patients experience recurring ischaemic episodes. Aspirin resistance denotes those situations when it is unable to protect individuals from thrombotic complications, or when it fails to produce an anticipated effect in laboratory tests of platelet function. There are various laboratory techniques with which to evaluate the effectiveness of aspirin and other antiplatelet drugs. It has been estimated that in 5-60% of patients, aspirin does not achieve adequate efficacy in various measures of platelet activity. Some studies have revealed that vascular patients shown by laboratory tests to be aspirin-resistant are at an increased risk of major vascular events. The suggested mechanisms of aspirin resistance, among others, include genetic polymorphisms, alternate pathways of platelet activation, aspirin-insensitive thromboxane biosynthesis, drug interactions, or low aspirin dose. An increase in the dosage of aspirin or conversion to clopidogrel or clopidogrel plus aspirin might be beneficial in the management of those patients who are aspirin resistant. Additional work is required to improve and validate laboratory tests of platelet function, so that they may become useful tools for selecting the most appropriate antiplatelet therapy for an individual patient. Improvements in antiplatelet treatment strategies in the future should lead to a reduction in premature vascular events. PMID- 15760637 TI - Neuroacanthocytosis: new developments in a neglected group of dementing disorders. AB - Neurological abnormalities associated with spiculated, "acanthocytic" red cells in blood have been summarized as neuroacanthocytosis. This is a heterogeneous group of conditions that can now be clearly subdivided on the basis of genetic discoveries. The core neuroacanthocytosis syndromes are autosomal recessive chorea-acanthocytosis (ChAc) and the X-linked McLeod syndrome (MLS). Huntington's disease-like 2 (HLD2) and pantothenate kinase associated neurodegeneration (PKAN) can now also be included. All of these share dyskinesias, cognitive deterioration and progressive neurodegeneration mainly of the basal ganglia, but they are sufficiently distinct to permit a specific working diagnosis on the basis of clinical, laboratory and imaging findings. In addition, the VPS13A (formerly called CHAC), XK, JPH3 and PANK2 genes, respectively, may be examined for mutations. Unfortunately, little is yet known about the normal and abnormal physiology of the protein products of these genes, but they appear to be involved in membrane function and intracellular protein sorting. Since no cures are yet available, development and study of disease models in experimental animals (mouse, C. elegans) is a priority for current research. From a clinical point of view, the common occurrence of cardiomyopathy in MLS, the transfusion hazards due to the McLeod Kell phenotype and the possibility of improving the violent trunk spasms and orofacial dyskinesias typical for ChAc (with subsequent lip or tongue mutilations and feeding dystonia) by deep brain surgery or stimulation should be considered in patient management. PMID- 15760639 TI - Neuropsychological evaluation of late-onset post-radiotherapy encephalopathy: a comparison with vascular dementia. AB - It is known that radiotherapy (RT) may cause cerebral injury. The most frequent neurotoxic effect of RT at any age is diffuse cerebral injury. Brain injury by therapeutic irradiation has traditionally been classified according to its time of onset into acute, early delayed, and late forms. The latter is not reversible. The neurocognitive sequelae of cranial irradiation can be mediated through vascular injury. Because the pathologic changes are most profound in the white matter, we compared a group of patients treated by RT (n=34) with a group of patients affected by subcortical vascular dementia (sVaD, n=34). Patients with a total radiation does <35 cGy did not show any sign of cognitive impairment. All the patients with a total irradiation dose >45 cGy did show profound cognitive and behavioural alteration. The patients who received a total dose of brain radiation comprised between 35 and 45 cGy did show slowness of executive function, and profound alterations of frontal functions, such as attention focusing, mentation control, analogical judgement and insight. The patients who suffered from the consequences of RT had slowness of executive functions, and profound alterations of frontal functions, such as attention focusing, mentation control, analogical judgement and insight, similar to those obtained by the patients suffering from subcortical vascular dementia. High dose RT might result in a severely demented, bedridden patient, who "has been cured" from his primary disease, the brain tumour. This constellation demands serious consideration before RT is given. PMID- 15760638 TI - Hypocitrullinemia in patients with MELAS: an insight into the "MELAS paradox". AB - L-citrulline, classified as a nonessential amino acid, is synthesized predominantly via Delta-1-pyrroline carboxylate synthase in the endothelial cells of the small intestine. In mammals, small quantities of citrulline are also produced in nitric oxide synthase-expressing cells. Considering the fact that the enzymes involved in the endogenous synthesis of L-citrulline are all located in the mitochondria and the fact that citrulline is a component of the citrulline nitric oxide (NO) cycle, we hypothesized that the distinct clinical, biochemical, and morphological characteristics of MELAS, a maternally inherited mitochondrial disorder, might be due to alterations in nitric oxide homeostasis. Analysis of serum from MELAS patients showed that levels of plasma arginine were similar in both patients and in controls. However, levels of citrulline in MELAS patients were significantly lower than in controls, and there was a clear inverse correlation between arginine and citrulline levels in these patients. We found no correlation between the level of heteroplasmy and the plasma levels of either arginine or citrulline. We discuss the depressed citrulline levels in MELAS patients, who have an unusual and paradoxical pattern of vascular respiratory chain expression, in the context of NO homeostasis. PMID- 15760640 TI - Understanding regulation of nerve cell death by mGluRs as a method for development of successful neuroprotective strategies. AB - A common cause of nerve cell death often leading to vascular dementia is ischemic stroke. Attempts to develop clinically effective stroke treatment and prevention strategies based on pharmacological manipulations of a single mechanism have not led to clinical success. Analysis of clinical neuroprotection trials suggests that combination treatments may be more effective. To identify optimal components for such treatment, N-methyl-d-aspartate receptor (NMDAR) activation-induced cell death in organotypic hippocampal preparations was studied as a model of neurodegeneration that occurs in association with stroke or vascular dementia. Pharmacological manipulation of metabotropic glutamate receptors mGluR1 and 5 resulted in significant reduction of nerve cell susceptibility to NMDA-induced injury, suggesting that these receptors may function as physiological regulators of neuronal vulnerability. cDNA microarray analysis of over 1000 brain-related genes performed after the neuroprotective activation of group I metabotropic glutamate receptors (mGluRs) revealed a complex pattern of activation and inactivation of seemingly unrelated genes responsible for regulation of neuronal excitability, inflammation, cell death pathways, cell adhesion and transcriptional activation. Combined pharmacological targeting of these processes may provide basis for clinical trials of effective neuroprotective compounds. PMID- 15760641 TI - Cholinesterase inhibitors exert a protective effect on endothelial damage in Alzheimer disease patients. AB - It has been recently suggested that in Alzheimer disease (AD), the current available therapy with cholinesterase inhibitors (ChEIs) influences platelets amyloid precursor protein (APP) metabolism towards the nonamyloidogenic pathway. In order to investigate whether ChEIs may exert a protective role on vascular damage due to abeta deposition, several parameters of coagulation and fibrinolysis were assessed. Twenty patients with mild AD and 30 age-matched controls entered the study. All subjects performed a multidimensional neuropsychological assessment and a laboratory protocol. Individuals with vascular risk factors and systemic diseases were excluded. In mild AD patients, increased levels of markers of endothelial dysfunction, such as thrombomodulin (TM) and sE-selectin (sE-sel), were seen. After 1-month ChEIs treatment, a significant reduction of TM (p<0.05) and sE-sel (p<0.05) values towards the normal range was observed. These findings suggest that endothelial-related ChEIs action might contribute to the clinical efficacy in AD, slowing down pathology progression. PMID- 15760642 TI - Anti-heparan sulphate antibodies and homocysteine in dementia: markers of vascular pathology? AB - Increasing evidence supports a pathogenic role of heparan sulphate (HS) in the development of dementia. Since HS proteoglycans are present in the endothelial cells and perivascular basement membrane, we wanted to assess blood titres of HS antibodies (Abs) in patients with vascular dementia (VD) and in patients with Alzheimer's disease (AD) with cerebrovascular disease (CVD) [mixed dementia (MixD)]. Moreover, plasma levels of homocysteine, an independent risk factor for the development of dementia as well as for CVD, were also determined. High HS Abs titres were present in one patient with VD and in two patients with mixed dementia, as well as in two neurological control patients (stroke and epilepsy). Increased homocysteine levels were found in 62.5% of patients with mixed dementia, in 22.2% of the VD subjects, in 54.2% of patients with CVD, and in 41.2% of patients with other neurological diseases. The present findings suggest that neither elevated HS Abs titres nor increased homocysteinemia may represent a useful biochemical marker for the diagnosis of VD. PMID- 15760643 TI - [11C]vinpocetine: a prospective peripheral benzodiazepine receptor ligand for primate PET studies. AB - Vinpocetine, a synthetic derivative of the Vinca minor alkaloid vincamine, is a widely used drug in neurological practice. We tested the hypothesis that vinpocetine binds to peripheral benzodiazepine binding sites (PBBS) and is therefore a potential ligand of PBBS. Positron emission tomography (PET) measurements in two cynomolgous monkeys showed that pretreatment with vinpocetine markedly reduced the brain uptake of [11C]PK11195, a known PBBS radioligand. On the other hand, whereas pretreatment with PK11195 increased the brain uptake of [11C]vinpocetine due to the blockade of PBBS in the periphery, it significantly reduced the binding potential (BP) values of [11C]vinpocetine in the whole brain and in individual brain structures to PK11195. These findings indicate that, whereas the two ligands have different affinities to PBBS, vinpocetine is a potent ligand of PBBS, which in turn suggests that the pharmacological activity of vinpocetine may involve the regulation of glial functions. PMID- 15760644 TI - Murine synaptosomal lipid raft protein and lipid composition are altered by expression of human apoE 3 and 4 and by increasing age. AB - Apolipoprotein E (apoE) 4 and aging are risk factors for Alzheimer's disease (AD). Mice expressing human apoE4 and aged wild-type mice show a similarity in the transbilayer distribution of cholesterol in synaptic plasma membranes (SPMs) but differ markedly compared with apoE3 mice and young mice. The largest changes in cholesterol distribution were observed in the SPM exofacial leaflet where there was a doubling of cholesterol. Lipid rafts are thought to be associated with the exofacial leaflet, and we proposed that lipid raft protein and lipid composition would be associated with apoE genotype and age. Lipid rafts were isolated from synaptosomes of different age groups (2, 12, 24 months) of mice expressing human apoE3 and apoE4. Lipid raft markers, alkaline phosphatase (ALP), flotillin-1, cholesterol and sphingomyelin (SM) were examined. Lipid rafts of young apoE4 mice were more similar to older mice as compared with young apoE3 mice in reductions in alkaline phosphatase activity and flotillin-1 abundance. Lipid raft cholesterol and sphingomyelin levels were not significantly different between the young apoE3 and apoE4 mice but cholesterol levels of lipid rafts did increase with age in both genotypes. Results of the present study demonstrate that the two risk factors for Alzheimer's disease, apoE4 genotype and increasing age have similar effects on brain lipid raft protein markers and these findings support the notion that the transbilayer distribution of cholesterol is associated with lipid raft function. PMID- 15760645 TI - Cholesterol homeostasis failure as a unifying cause of synaptic degeneration. AB - We previously showed that fine tuning of neural cholesterol dynamics is essential for basic synapse function, plasticity and behavior. Significant experimental evidence indicates that cholinergic function, ionotropic and metabotropic receptor machinery, excessive tau phosphorylation, the change of amyloid beta (Abeta or Abeta) biochemistry, neural oxidative stress reactions, and other features of neurodegeneration also depend on fine tuning of brain cholesterol homeostasis. This evidence suggest that (i) cholesterol homeostasis break is the unifying primary cause of sporadic and familial Alzheimer's disease (AD), neuromuscular diseases (particularly inclusion-body myositis), Niemann-Pick's type C disease and Down syndrome, and (ii) explains the overlap of neurodegenerative hallmarks across the spectrum of neurodegenerative diseases. Provided is evidence-based explanation of why extremely rare (but scientifically popular) cases of AD associated with mutations in amyloid beta protein precursor (APP) and presenilin (PS) genes, are translated into the disorder via membrane cholesterol sensitivity of APP processing by secretases and Abeta generation. The reciprocal effect of Abeta on cholesterol synthesis, cellular uptake, efflux and esterification is summarized, as well as the potential implication of such biological function for the compensatory Abeta-assisted restoration of the synaptic long-term potentiation (LTP) and resulting inability of tackling amyloid to cure AD. PMID- 15760646 TI - Brain area- and isoform-specific inhibition of synaptic plasticity by apoE4. AB - The allele E4 of apolipoprotein E4 (apoE4), the most prevalent genetic risk factor of Alzheimer's disease (AD), inhibits the improvements in learning and memory which result from exposure of apoE transgenic mice to environmental stimulation (ES). In the present study, we investigated the extent to which these cognitive deficits are associated with distinct presynaptic, postsynaptic and axonal impairments and whether these effects are brain area-specific. Exposure to an enriched environment of young mice transgenic for human apoE3, which is the AD benign apoE allele, increased the levels of the presynaptic protein synaptophysin and of the dendritic marker MAP-2 in the hippocampus and entorhinal cortex, whereas the corresponding levels of these proteins in the apoE4 transgenic mice were unaffected by the enriched environment. In contrast, the levels of synaptophysin and MAP-2 in the motor cortex were elevated by environmental stimulation in both the apoE3 and the apoE4 transgenic mice. These findings show that apoE4 inhibits synaptic plasticity following environmental stimulation and that this effect is both isoform- and brain area-specific. PMID- 15760647 TI - Matrix metalloproteinase-9 in cerebral-amyloid-angiopathy-related hemorrhage. AB - Spontaneous intracerebral hemorrhage (ICH) is one of the most recognized complications of cerebral amyloid angiopathy (CAA), but little is known about the molecular pathogenesis of this life-threatening complication. In this review, we present preliminary evidence which suggests that the extracellular-matrix degrading protease, matrix metalloproteinase-9 (MMP-9), may play a role in the development of spontaneous ICH resulting from CAA. The amyloid-beta peptide (Abeta) induced the synthesis, cellular release, and activation of MMP-9 in murine cerebral endothelial cells (CECs), resulting in increased extracellular matrix (ECM) degradation. Furthermore, in a mouse model of CAA (APPsw transgenic mice), MMP-9 immunoreactivity was observed in amyloid-laden cerebral vessels in aged APPsw mice but not in young APPsw or aged wild-type mice. More extensive MMP 9 immunostaining was present in amyloid-laden vessels with evidence of microhemorrhage. These results suggest that increased vascular MMP-9 expression, stimulated by Abeta, may play a role in the pathogenesis of spontaneous intracerebral hemorrhage (ICH) in patients with CAA. PMID- 15760648 TI - Non-human primate SPECT model for determining cerebral perfusion effects of cerebrovasoactive drugs acting via multiple modes of pharmacological action. AB - Increasing clinical and experimental evidence implicate cerebral hypoperfusion during increased ageing and points to chronic cerebrovascular ischemia as a vital component of the neuropathological progression of dementia. In vivo cerebral perfusion animal models can greatly contribute to the evaluation of drugs and to the screening of drug interactions. This study describes a baboon Papio ursinus model under anaesthesia, for in vivo cerebral blood flow (CBF) determinations, using Single Photon Emission Computed Tomography (SPECT) following the split-dose method with 99mTc-hexamethylpropylene amine oxime (99mTc-HMPAO). Perfusion studies with acetazolamide as intervention clearly showed that the non-human primate model under aneasthesia is sufficiently sensitive to serve in the evaluation of other cerebrovasoactive drugs for induced perfusion changes with significant increases of the R-value (+40%) for comparative measurement when compared to the control value (2.53+/-0.15 vs. 1.79+/-0.13). These findings stimulated investigations of several drugs, i.e. pentifylline (phosphodiesterase inhibitor); nimodipine (calcium channel blocker); sumatriptan (serotonin receptor agonist) and nicotinic acid (vasodilator) for CBF effects. Increases in the cerebral perfusion in some cases more than +30% for nimodipine (2.51+/-0.14 vs. 1.79+/-0.13), acetazolamide and +29% for the combination of pentifylline and nicotinic acid (2.31+/-0.19 vs. 1.79+/-0.13) were observed. Drug interaction studies revealed an attenuation of increased CBF due to nimodipine, with sumatriptan (-25%) and acetazolamide (+22%) in combination with nimodipine. Drug interactions with clinical implications may result during simultaneous use of cerebrovasoactive drugs in managing patients with cerebrovascular disorders. This study further showed that the CBF non-human primate model under anaesthesia is useful for the investigation of vasoactive drugs acting via various pharmacological modes of action. PMID- 15760649 TI - Differential and quantitative molecular analysis of ischemia complexity reduction by isotopic labeling of proteins using a neural embryonic stem cell model. AB - The analysis of rapid changes of protein expression in living systems in response to insults requires rigorous methods of complexity reduction. To control dynamic pattern of hundreds or even thousands of protein isoforms, we applied a novel method of differential molecular analysis to a cellular model which is suited to study ischemia. Neural derivatives of murine embryonic stem cells were exposed to chemical ischemia. The model was used to obtain starting material for a quantitative differential proteomics analysis. Fractionation of phosphoproteins from these samples and subsequent identification by mass spectrometry of differential proteins provide proof of principle of how novel molecular analytical tools provide new insight into the network of neuroprotective molecular events during specific situations of neuronal stress and related pharmaceutical intervention. Our results indicate a particular role of an isoform of the acidic calcium-independent phospholipase A2 in this type of insult. PMID- 15760650 TI - The regulation of beta-secretase by cholesterol and statins in Alzheimer's disease. AB - Epidemiologists have found a decreased risk of developing Alzheimer's disease (AD) in people taking statins (cholesterol biosynthesis inhibitors). We have reported previously that, in cell culture, lovastatin decreases the output of beta-amyloid, a peptide that is toxic to neurones, and is reputably the prime cause of neurodegeneration seen in AD. This report probes the mechanism of statin protection further by finding out how the protease beta-secretase, that releases beta-amyloid from its precursor protein, behaves under changed cholesterol levels induced by statins. We found that, with high cellular cholesterol levels, there is a decrease in glycosylation of mature oligosaccharides in beta-secretase, whereas in the presence of lovastatin, glycosylation progresses further. Moreover, lovastatin does not inhibit beta-secretase in vitro. Thus, the cholesterol and statin effects are due to changes in cellular targeting induced by changed cholesterol gradients. Some of these changes are mimicked by the action of U18666A, a cholesterol-transport inhibitor that produces a defect in cells seen in patients with Neimann Pick's disorder. PMID- 15760651 TI - Effects of vinpocetine on the redistribution of cerebral blood flow and glucose metabolism in chronic ischemic stroke patients: a PET study. AB - The pharmacological effects of the neuroprotective drug vinpocetine, administered intravenously in a 14-day long treatment regime, on the cerebral blood flow and cerebral glucose metabolism in chronic ischemic stroke patients (n=13) were studied with positron emission tomography in a double-blind design. The regional and global cerebral metabolic rates of glucose (CMRglc) and cerebral blood flow (CBF) as well as vital physiological parameters, clinical performance scales, and transcranial Doppler parameters were measured before and after the treatment period in patient groups treated with daily intravenous infusion with or without vinpocetine. While the global CMRglc values did not change markedly as a result of the infusion treatment with (n=6) or without (n=7) vinpocetine, the global CBF increased and regional CMRglc and CBF values showed marked changes in several brain structures in both cases, with more accentuated changes when the infusion contained vinpocetine. In the latter case the highest rCBF changes were observed in those structures in which the highest regional uptake of labelled vinpocetine was measured in other PET studies (thalamus and caudate nucleus: increases amounting to 36% and 37%, respectively). The findings indicate that a 2-week long intravenous vinpocetine treatment can contribute effectively to the redistribution of rCBF in chronic ischemic stroke patients. The effects are most pronounced in those brain regions with the highest uptake of the drug. PMID- 15760652 TI - Mitochondria DNA deletions in atherosclerotic hypoperfused brain microvessels as a primary target for the development of Alzheimer's disease. AB - The pathogenesis, which is primarily responsible for Alzheimer's disease (AD) and cerebrovascular accidents (CVA), seems to involve chronic hypoperfusion. The role of hypoperfusion, as a key factor for vascular lesions that causes oxidative stress, appears to be widely accepted as an initiator of AD. Specifically, accumulated oxidative stress increases vascular endothelial permeability and promotes leukocyte adhesions, which is coupled with alterations in endothelial signal transduction and redox-regulated transcription factors. Based on these recent findings, we hypothesize that the cellular and molecular mechanisms by which hypoperfusion-induced reactive oxygen species (ROS) accumulation impairs endothelial barrier function and promotes leukocyte adhesion induces alterations in normal vascular function and results in the development of AD. We are theorizing that mitochondria play a key role in the generation of ROS, resulting in oxidative damage to neuronal cell bodies, as well as other cellular compartment in the AD brain. All of these changes have been found to accompany AD pathology. We have studied the ultrastructural features of vascular lesions and mitochondria in brain vascular wall cells from human AD, yeast artificial chromosome (YAC) and C57B6/SJL transgenic positive (Tg+) mice overexpressing amyloid beta precursor protein (AbetaPP). In situ hybridization using mitochondrial DNA (mtDNA) probes for human wild and 5 kb deleted types and mouse types was performed along with immunocytochemistry using antibodies against amyloid precursor protein (APP), 8-hydroxy-2'-guanosine (8-OHG) and cytochrome c oxidase (COX). There was a higher degree of amyloid deposition, overexpression of oxidative stress markers, mitochondria DNA deletion and mitochondrial structural abnormality in the vascular walls of the human AD, YAC and C57B6/SJL Tg (+) mice compared to age-matched controls. Therefore, selective pharmacological intervention, directed for abolishing the chronic hypoperfusion state, would possibly change the natural course of development of dementing neurodegeneration. PMID- 15760653 TI - An in vivo role for Trypanosoma cruzi calreticulin in antiangiogenesis. AB - Angiogenesis leads to neovascularization from existing blood vessels. It is associated with tumor growth and metastasis and is regulated by pro- and antiangiogenic molecules, some of them currently under clinical trials for cancer treatment. During the last few years we have cloned, sequenced and expressed a Trypanosoma cruzi calreticulin gene (TcCRT). Its product, TcCRT, a 45 kDa protein, is more than 50% identical to human CRT (HuCRT). TcCRT, present on the surface of trypomastigotes, binds both C1q and mannan binding lectin and inhibits the classical activation pathway of human complement. Since TcCRT is highly homologous to a functional antiangiogenic fragment from HuCRT (aa 120-180), recombinant (r) and native (n) TcCRT were tested in their antiangiogenic effects, in the chick embryonic chorioallantoid membrane (CAM) assay. Both proteins mediated highly significant antiangiogenic effects in the in vivo CAM assay. This effect was further substantiated in experiments showing that the plasmid construct pSecTag/TcCRT also displayed significant antiangiogenic properties, as compared to the empty vector. Most likely, the fact that antiangiogenic substances act preferentially on growing neoplasic tissues, but not on already established tumors, is due to their effects on emerging blood vessels. The results shown here indicate that TcCRT, like its human counterpart, has antiangiogenic properties. These properties may explain, at least partly, the reported antineoplasic effect of experimental T. cruzi infection. PMID- 15760654 TI - Phylogenetic relationships of methionine aminopeptidase 2 among Encephalitozoon species and genotypes of microsporidia. AB - This report describes the characterization and phylogenetic analysis of the deduced amino acid sequences of methionine aminopeptidase 2 (MetAP-2) enzymes from microsporidian species and genotypes of the genus Encephalitozoon. Fragments of DNA encoding 318 to 335 amino acid residues of the MetAP-2 genes were isolated from genomic DNA prepared from cultured spores of Encephalitozoon hellem, Encephalitozoon intestinalis, and Encephalitozoon cuniculi genotypes I-III. Sequence comparisons of the deduced amino acid residues indicated that the microsporidian sequences are MetAP-2-like rather than MetAP-1-like. Alignments demonstrated that the new Encephalitozoon sequences included sequences and structures conserved in eukaryotic MetAP-2s, including the five conserved, active site residues, Asp, Asp, His, Glu, and His, considered to be critical for catalysis and for coordinating the cation (e.g., cobalt) co-factor, and included residues known to interact with the antibiotic, fumagillin. The primary structure of the Encephalitozoon MetAP-2s, however, showed some dissimilarity with human and yeast MetAP-2s, including the absence of the NH2-terminal polylysine tract. Phylogenetic comparison of these Encephalitozoon MetAP-2s with orthologues from related species and from other informative taxa confirmed that the MetAP-2s of these Encephalitozoon species and strains are closely related to each other and cluster with MetAP-2s. PMID- 15760655 TI - Implication of a Plasmodium falciparum gene in the switch between asexual reproduction and gametocytogenesis. AB - Gametocytogenesis is fundamental for transmission of the malaria parasite Plasmodium falciparum from the human host to the mosquito vector, yet very little is understood about what triggers the switch between asexual reproduction and gametocytogenesis. Arresting the progression through the sexual cycle would block transmission of this disease. Here we identify a novel gene in P. falciparum that when genetically silenced reduces gametocyte production by a factor of 6, and when complemented up-regulates gametocyte-specific gene transcription. PMID- 15760656 TI - A unique Rab GTPase, EhRabA, is involved in motility and polarization of Entamoeba histolytica cells. AB - Entamoeba histolytica, an enteric protozoan parasite, infects 10% of the world's population leading to 50 million cases of invasive amoebiasis annually. Motility, which requires cell polarization, is important to the virulence of this pathogen, as it may result in destruction of host tissues and invasion. To gain insight into these processes in Entamoeba, a unique Rab GTPase, EhRabA, which localizes to the leading edge of cells, was characterized. Cell lines expressing a dominant negative version of EhRabA (EhRabA-DN) were generated. These mutant cells exhibited alterations in cell shape, polarity, and motility, supporting a role for this Rab in the regulation of these processes. Consistent with the notion that a dynamic actin cytoskeleton is crucial to cell polarity and motility, these mutants also exhibited alterations in the actin cytoskeleton. Cells expressing EhRabA-DN also displayed defects in several virulence functions including the ability to adhere to host cells, destroy host cells, and release cysteine proteases. Mislocalization of a prominent adhesion molecule, the galactose/N acetylgalactosamine (Gal/GalNAc) adherence lectin and reorganization of ordered lipid domains, known as lipid rafts, also accompanied expression of EhRabA-DN. Interestingly, several endocytic processes were unaffected by expression of EhRabA-DN. Together, these data suggest that EhRabA may be involved in the regulation of polarization, motility and actin cytoskeletal dynamics: functions that participate in the pathogenicity of Entamoeba. PMID- 15760657 TI - A proton pumping pyrophosphatase in acidocalcisomes of Herpetomonas sp. AB - Acidocalcisomes are acidic calcium storage organelles found in several microorganisms. They are characterized by their acidic nature, high electron density, high content of polyphosphates and several cations. Electron microscopy contrast tuned images of Herpetomonas sp. showed the presence of several electron dense organelles ranging from 100 to 300 nm in size. In addition, X-ray element mapping associated with energy-filtering transmission electron microscopy showed that most of the cations, namely Na, Mg, P, K, Fe and Zn, are located in their matrix. Using acridine orange as an indicator dye, a pyrophosphate-driven H+ uptake was measured in cells permeabilized by digitonin. This uptake has an optimal pH of 6.5-6.7 and was inhibited by sodium fluoride (NaF) and imidodiphosphate (IDP), two H+-pyrophosphatase inhibitors. H+ uptake was not promoted by ATP. Addition of 50 microM Ca2+ induced the release of H+, suggesting the presence of a Ca2+/H+ countertransport system in the membranes of the acidic compartments. Na+ was unable to release protons from the organelles. The pyrophosphate-dependent H+ uptake was dependent of ion K+ and inhibited by Na+ Herpetomonas sp. immunolabeled with monoclonal antibodies raised against a Trypanosoma cruzi V-H+-pyrophosphatase shows intense fluorescence in cytoplasmatic organelles of size and distribution similar to the electron-dense vacuoles. Together, these results suggest that the electron dense organelles found in Herpetomonas sp. are homologous to the acidocalcisomes described in other trypanosomatids. They possess a vacuolar H+-pyrophosphatase and a Ca2+/H+ antiport. However, in contrast to the other trypanosomatids so far studied, we were not able to measure any ATP promoted H+ transport in the acidocalcisomes of this parasite. PMID- 15760658 TI - Recombinant and native Plasmodium falciparum TATA-binding-protein binds to a specific TATA box element in promoter regions. AB - RNA polymerase II promoters in Plasmodium spp., like in most eukaryotes, have a bipartite structure. However, the identification of a functional TATA box located within the Plasmodium spp. core promoters has been difficult, mainly because of its high A+T content. Only few putative trans-acting elements have been identified in the malaria parasite genome such as a gene orthologous to the TATA box binding protein (PfTBP). In this study, we demonstrate that PfTBP is part of the DNA-protein complexes formed in the kahrp and gbp-130 gene promoter regions. Supershift and footprinting assays performed with a GST-PfTBP fusion protein showed that PfTBP associates with a consensus TATA box sequence located 81 base pairs upstream of the transcription start site in the kahrp promoter region and with a TATA box-like (TGTAA) sequence at position -186 of the gbp-130 gene promoter region. Chromatin immunoprecipitation assays confirmed that native PfTBP is able to associate in vivo with both TATA box elements. This is the first study that reports the identification of cis-acting sequences (TATAA and TGTAA) and their corresponding trans-acting (PfTBP) factor in P. falciparum. PMID- 15760659 TI - Molecular and electrophysiological characterization of nucleotide-sensitive chloride current-inducing protein of Fasciola hepatica. AB - Nucleotide-sensitive chloride current regulating proteins (ICln's) of the chloride channels have been characterized from man and animals. An ICln of Fasciola hepatica (ICln-Fh) consisting of 231 amino acids revealed high similarities to both consensus domain of ICln's and two acidic residue-abundant patches in its C-terminus. Native ICln-Fh protein was confirmed present in F. hepatica soluble extract by immunoblotting. The recombinant ICln-Fh protein expressed in collagenase-defolliculated Xenopus oocytes induced fast rising and outward rectifying Cl- currents (I(Cln-Fh)). The recombinant ICln-Fh protein, however, did not trigger cell swelling-induced Cl- currents (I(Cl-swell)). The I(Cln-Fh) currents were significantly reduced by substituting external Cl- with gluconic acid and by externally adding cAMP. Collectively, these results suggest that ICln-Fh protein is an inducer of Cl- currents in F. hepatica. PMID- 15760660 TI - Characterization and developmental gene regulation of a large gene family encoding amastin surface proteins in Leishmania spp. AB - The ability of Leishmania amastigotes to survive within the drastic environmental changes encountered in the phagolysosomes of mammalian macrophages is heavily dependent on the developmental regulation of a variety of genes. The identification of genes that are expressed preferentially in the mammalian stage of the parasite should increase our understanding of the molecular mechanisms regulating stage-specific gene expression and of the determinants that control its intracellular survival and contribute to its pathogenesis. We report here detailed sequence characterization and structural organization of the amastin gene family in Leishmania major and Leishmania infantum and the study of their developmental gene regulation throughout the parasite's life cycle. Amastin surface proteins represent the largest developmentally regulated gene family reported so far in Leishmania comprising up to 45 members. All the members of the amastin gene family in both Leishmania and Trypanosoma species share a similar structural organization and contain a highly conserved 11 amino acid extracellular domain, which is unique to amastin proteins. The majority of the amastin gene homologs are specifically expressed in the amastigote stage of the parasite. Three distinct RNA elements were identified in the 3'-untranslated regions (3'UTR) of the amastin transcripts. The majority of these transcripts contain a conserved 450 nt cis-acting 3'UTR element shown previously to regulate stage-specific gene expression at the level of translation, which suggests that several amastin homologs may be regulated by a similar mechanism of translational control inside the macrophage. These findings further highlight the unique features of gene expression control in Leishmania. PMID- 15760661 TI - Comparative analysis of genomic sequences suggests that Trypanosoma cruzi CL Brener contains two sets of non-intercalated repeats of satellite DNA that correspond to T. cruzi I and T. cruzi II types. AB - Approximately 10% of the Trypanosoma cruzi genome is formed by a satellite DNA, composed by 195-bp repeats organized in 30+/-10 kb clusters in some, but not all chromosomes. Here, the satellite DNA of six representative T. cruzi strains was sequenced and used for phylogenetic inference. The results show that CL Brener contains satellite repeats from T. cruzi I and T. cruzi II strains, although type II sequences are more abundant. The presence of types I and II sequences extends previous propositions that genetic exchange between the two major T. cruzi lineages have occurred in CL Brener, although our data accommodate alternative scenarios of hybridization within T. cruzi II, as proposed by others. Altogether, present data suggest a complex origin for CL Brener. Sequence analysis of satellites isolated from chromosomal bands indicates that satellite DNA sequences are not chromosome specific. Neighbor analysis of in tandem satellite DNAs containing up to five repeats shows that each cluster contains only one type of sequence. Consequently, clusters with intercalated types I and II repeats were not found. We propose that the CL Brener genome contains large pieces of satellite DNA originated mainly from chromosomes of T. cruzi II with introgression of T. cruzi I lineage. PMID- 15760662 TI - Population genetic analysis and sub-structuring of Theileria parva in Uganda. AB - In recent years the population structures of many apicomplexan parasites including Plasmodium spp., Toxoplasma gondii and Cryptospordium parvum have been elucidated. These species show a considerable diversity of population structure suggesting different strategies for transmission and survival in mammalian hosts. We have undertaken a population genetic analysis of another apicomplexan species (Theileria parva) to investigate the levels of diversity of this parasite and the role of genetic exchange in three geographically separate populations. The principal hindrance to carrying out such a study on field isolates was the high proportion of blood samples that contain multiple genotypes, making it impossible to determine the genotypes of the parasites directly. This problem was overcome by sampling only young indigenous calves between 3 and 9 months of age in which approximately 60% of the T. parva infected calves contained a single/predominant allele at each locus, making it possible to undertake population genetic analyses. Blood samples were collected from calves in three geographically distinct regions of Uganda and were analysed using 12 polymorphic mini and microsatellite markers that were evenly dispersed across the four chromosomes. We have identified 84 multilocus genotypes (MLG) from these samples, indicating high levels of diversity in the parasite. Analysis of linkage disequilibrium between pairs of loci provides evidence that the population in Lira district had an epidemic structure. The population in Mbarara was substructured containing two genetically distinct sub-groups and the larger sub-group also had an epidemic population structure. The population from Kayunga was in linkage disequilibrium. Genetic distances and Wrights fixation index (F(ST)) indicate that there is evidence for geographical sub-structuring between the Lira and the Kayunga populations. PMID- 15760663 TI - An analysis of genetic diversity within the ligand domains of the Plasmodium falciparum ebl-1 gene. PMID- 15760664 TI - Tetracycline induction of gene expression in Trypanosoma brucei within the tsetse fly vector. PMID- 15760666 TI - Epigallocatechin-3-gallate from green tea negatively affects swine granulosa cell function. AB - The use of herbs as additives in livestock nutrition as an alternative to antibiotics is becoming a new goal in animal production. It is known that green tea exerts antimicrobial activity owing to specific flavonoid compounds named catechins, primarily represented by epigallocatechin-3-gallate (EGCG). Remarkably, despite many potential benefits of green tea and EGCG consumption, it is also important to get an insight on the possible reproductive-related consequences of feeding supplementation. To this purpose, granulosa cells were harvested from follicles > 5mm and treated with 5 and 50 microg/ml of EGCG in order to evaluate the effects on the main parameters of granulosa cell function: steroidogenesis, by measuring progesterone and estradiol-17beta production, and proliferation, one of the major feature of ovarian follicular growth. Moreover, as the genesis of new vessels has been demonstrated to be fundamental for follicle development, we evaluated the effect of EGCG on the production of the main angiogenetic factor, VEGF, by swine granulosa cells. Finally, since reactive oxygen species (ROS) might be involved in the control of female reproductive activity, we studied the effect of EGCG on superoxide anion (O2-) and hydrogen peroxide (H2O2) production by swine granulosa cells and on the activity of the scavenging enzyme superoxide dismutase (SOD). EGCG significantly (p < 0.05) inhibited proliferation, steroidogenesis, VEGF and O2- production by swine granulosa cells; on the contrary, H2O2 levels and SOD activity were stimulated (p < 0.05) by the catechin. Therefore, since our data demonstrate that EGCG has a negative effect on reproductive performances in swine, feeding supplementation should be carefully considered. PMID- 15760665 TI - Regulation of in vitro growth of preantral follicles by growth factors in goats. AB - Goat preantral follicles were cultured to investigate the effects of insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on the in vitro growth and viability of oocytes. Preantral follicles were isolated mechanically and enzymatically (using collagenase and DNase) from prepuberal goat ovaries. The working medium was composed of Defined Eagle's Minimum Essential Medium (DMEM) supplemented with HEPES (20 mM), 10% fetal calf serum (FCS), hypoxanthine (2 mM), dibutyryl cyclic adenosine 3',5' monophosphate (dbcAMP) (2 mM), penicillin (75 ng/ml) and streptomycin (50 ng/ml). The culture medium consisted of the working medium with follicle stimulating hormone (FSH) (100 ng/ml) and hydrocortisone (40 ng/ml) added. In the experiment, goat preantral follicles were cultured for 9 days in the culture medium and in the culture medium supplemented with either IGF-I (100 ng/ml), EGF (50 ng/ml), bFGF (50 ng/ml) or IGF-I (100 ng/ml)+EGF (50 ng/ml). The results indicated that IGF-I (100 ng/ml) effectively maintained the survival of oocytes and promoted their growth; EGF (50 ng/ml) enhanced the survival rate of oocytes but had a negative effect on oocyte growth; bFGF (50 ng/ml) stimulated oocyte survival but had no obvious effect on their growth while IGF-I (100 ng/ml) and EGF (50 ng/ml) in combination had a greater effect on both survival and growth rate of oocytes than IGF-I or EGF alone. The supplementation of IGF-1 and EGF to the culture medium is recommended in the culture of goat preantral follicles. PMID- 15760668 TI - Fasting influences steroidogenesis, vascular endothelial growth factor (VEGF) levels and mRNAs expression for VEGF, VEGF receptor type 2 (VEGFR-2), endothelin 1 (ET-1), endothelin receptor type A (ET-A) and endothelin converting enzyme-1 (ECE-1) in newly formed pig corpora lutea. AB - This study was designed to verify whether fasting influences vascular endothelial growth factor (VEGF) production and VEGF, VEGF receptor-2 (VEGFR-2) as well as endothelin (ET) system members (endothelin converting enzyme-1, ECE-1; ET-1; endothelin receptor type A, ET-A) mRNA expression in pig corpora lutea; furthermore, we wanted to assess whether fasting affects steroidogenesis in luteal cells. Eight prepubertal gilts were induced to ovulate and were randomly assigned to two groups: (A) n = 4, normally fed; and (B) n = 4, fasted for 72 h starting 3 days after ovulation. At the end of fasting, ovaries were removed from all the animals and corpora lutea (CLs) were collected. VEGF and steroid levels in luteal tissue were determined by ELISA and RIA, respectively; VEGF, VEGFR-2, ET-1, ET-A and ECE-1 mRNAs expression was measured by real-time PCR. VEGF protein levels were similar in the two groups, while all steroid (progesterone, testosterone, estradiol 17beta) concentrations were significantly (P < 0.001) higher in CLs collected from fasted animals compared with those from normally fed gilts. VEGF, VEGFR-2, ET-1 and ECE-1 (but not ET-A) mRNA expression was significantly lower (P < 0.05) in fasted versus normally fed animals. The overall conclusion is that all the parameters studied are affected by feed restriction, but the mechanisms activated at luteal level are possibly not fully adequate to compensate for nutrient shortage. PMID- 15760667 TI - Effects of dexamethasone and colostrum feeding on mRNA levels and binding capacities of beta-adrenergic receptors in the liver of neonatal calves. AB - Glucocorticoids increase plasma glucose concentrations in neonatal calves, but not hepatic gluconeogenic enzyme mRNA levels and activities. Catecholamines, too, enhance plasma glucose levels and regulate hepatic glucose metabolism. We have measured hepatic mRNA levels of beta-adrenergic receptors and beta-adrenergic receptor binding in neonatal calves on day 5 of life. Calves were fed either colostrums (C) or an isoenergetic milk-based formula (F), and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/(kg body weightday)). Abundance of mRNA was highest (P < 0.01) for beta2-adrenergic receptors and was higher (P < 0.01) for beta1- than for beta3-adrenergic receptors. DEXA treatment decreased (P < 0.05) beta1- and beta2-adrenergic receptor mRNA levels. Beta3-adrenergic receptor mRNA levels were higher (P < 0.05) in colostrum- than in formula-fed calves. Competitive binding revealed highest affinities for alprenolol, propranolol (both beta1- and beta2 antagonists), and ICI-188,551 (beta2-antagonist), which did not significantly differ from each other. Atenolol (beta1-antagonist) up to 10(-5) M did not displace (3H)-CGP-12177 from receptors. Competitive binding for adrenaline was best fitted by a two-receptor model. DEXA decreased (P < 0.05) (3H)-CGP-12177 binding capacities, whereas binding affinity of (3H)-CGP-12177 was not affected by DEXA or different feeding. Binding sites correlated positively with mRNA levels of beta2-adrenergic receptors (r = 0.56; P < 0.01). In conclusion, beta2 adrenergic receptors were the dominant subtype in the hepatic tissue. Feeding did not significantly affect beta2-adrenergic binding sites. However, DEXA decreased beta2-adrenergic binding sites and this was regulated at the transcriptional level. PMID- 15760669 TI - The induction of a delayed post-ovulatory progesterone rise in dairy cows: a novel model. AB - A delayed rise in post-ovulatory progesterone is associated with poor embryo development in the cow, although the underlying cause of this aberrant luteal function is poorly understood. The objective of this study was to develop a novel model, in which a delayed progesterone rise could be induced by manipulating the dynamics of the follicular phase. Luteolysis was induced in 20 dairy cows in the presence of either a larger follicle > 10 mm (LF, n = 11) or a smaller follicle < 10 mm (SF, n = 9) and transrectal ultrasonography was performed to determine follicle and CL growth and timing of ovulation. Plasma progesterone and oestradiol were analysed 3x daily. Cows were slaughtered on either day 4 (n = 4 per group) or day 7 (SF, n = 5; LF, n = 7) after ovulation. The pre-ovulatory follicle was larger in the LF group than the SF group at luteolysis (13.5 +/- 0.4 mm versus 6.7 +/- 0.7 mm, P < 0.001) and ovulation (16.7 +/- 0.3 mm versus 13.6 +/- 0.6 mm, P < 0.001). The LF group experienced a shorter follicular phase and ovulated 36 h earlier than the SF group (P < 0.001). At luteolysis, plasma oestradiol concentrations were greater in the LF group (P < 0.001), although peak concentrations were not different (P > 0.05). Moreover, higher progesterone concentrations were observed in the LF group during the early luteal phase (P < 0.05). Luteal weights were positively correlated with plasma progesterone concentrations on day 5 (P < 0.05) but not day 8. In conclusion, a model has been developed which has shown that the dynamics of follicle development during the pre-ovulatory period is an important determinant of subsequent CL development and function. PMID- 15760670 TI - Correlation of blood and milk components with the milk yield response to bovine somatotropin in dairy cows. AB - Our aim was to correlate the individual variation in the milk yield response (MYR) of Holstein dairy cows to bovine somatotropin (bST), with changes in milk plasmin and plasminogen activities as well as with plasma hormone and metabolite levels. Thirty-two housed multiparous Holstein cows (90 +/- 3.8 days post partum) received daily subcutaneous injections of saline for 1 week followed by subcutaneous injections of 20 mg/day of bST for 2 weeks. Blood samples were taken at approximately 4h intervals over 24 h at the end of the saline and bST treatment periods. Milk samples were also taken at the end of the saline and bST treatment periods. The difference in milk yield between the saline and the second week of bST treatment (MYR) varied considerably between animals (from -0.2 to +8.6 kg/day, relative to the saline treatment week). Low milk yield before bST treatment was associated with a high MYR. The plasma growth hormone response to treatment was negatively correlated with MYR. Plasma insulin-like growth factor-1 response to treatment was positively correlated with MYR. Furthermore, a high MYR to bST was associated with a lower milk plasminogen level before treatment and a greater reduction in the level of plasminogen in milk following treatment. PMID- 15760671 TI - Angiogenic activity of swine granulosa cells: effects of hypoxia and vascular endothelial growth factor Trap R1R2, a VEGF blocker. AB - The possible role played by hypoxia and vascular endothelial growth factor (VEGF) in the regulation of follicular angiogenesis was studied in a three-dimensional fibrin gel model. Granulosa cells from follicles >5mm were subjected to normoxia (19% O2), partial (5% O2) or total (1% O2) hypoxia and their culture media were collected and used to stimulate porcine Aortic Endothelial Cells (AOC) included in the fibrin matrix. A suspension of AOC on microcarrier beads was pipetted in a fibrinogen solution (1 mg/ml PBS) before the addition of 1250 IU thrombine (250 microl) to catalize the gel formation. Granulosa cell conditioned media were tested in the presence or absence of VEGF Trap R1R2 (150 ng/ml), a potent VEGF inhibitor, that had its efficacy tested by adding VEGF (100 ng/ml) to AOC culture. Endothelial cell proliferation was measured at 48, 96, 144, 192 h by means of Scion Image Beta. A significant (p < 0.01) increase of AOC proliferation at each time of measurement was induced by culture media from granulosa cells subjected to partial (except at the end of the first 48 h) and total hypoxia compared to control and normoxia conditions, and by VEGF. VEGF Trap significantly (p < 0.01) inhibited the stimulatory effect of media conditioned by granulosa cells cultured in hypoxic conditions. These data suggest that hypoxia stimulates angiogenic activity of granulosa cells possibly by means of VEGF which could represent the main effector in promoting endothelial cell proliferation. PMID- 15760672 TI - Ontogenetic development of mRNA levels and binding sites of hepatic beta adrenergic receptors in cattle. AB - Catecholamines regulate glucose metabolism and affect hepatic glucose production mainly through beta2-adrenergic receptors. The hypothesis was tested that gene expression and numbers of hepatic beta-adrenergic receptors in calves are influenced by age. Examined developmental stages included pre-term (P0) and full term (F0) calves immediately after birth, full-term calves on day 5 of life (F5), and veal calves (VC) at the age of 159 days. Expression of beta1-, beta2-, and beta3-adrenergic receptor mRNA was measured by real-time PCR. Receptor binding was quantified by saturation binding assays using (3H)-CGP-12177 as a ligand. Abundance of mRNA differed among beta-adrenergic subtypes (beta2 > beta1 > beta3; P < 0.01). Beta3-adrenergic receptor mRNA was undetectable in VC. mRNA abundance for beta2-adrenergic receptors was higher (P < 0.05) in VC than P0 and for beta3 adrenergic receptors was higher (P < 0.001) in F5 than P0. Binding studies revealed most binding of (3H)-CGP-12177 to beta2-adrenergic receptors, which were highest in VC (P < 0.001) and higher (P < 0.05) in F5 than P0. Binding sites correlated positively with mRNA levels of beta2-adrenergic receptors (r = 0.67; P < 0.001), with hepatic activities of phosphoenolpyruvate kinase (r = 0.73; P < 0.001) and with pyruvate kinase (r = 0.4; P < 0.05), and with plasma glucose concentrations (r = 0.5; P < 0.01). In conclusion, mRNA of all three beta adrenergic receptor subtypes were found in liver, with beta2-adrenergic receptors being the dominant subtype. Numbers of beta2-adrenergic receptors increased with age and were mainly regulated at the transcriptional level. Numbers of beta adrenergic receptors were positively associated with hepatic activities of gluconeogenetic enzymes and with plasma glucose levels, suggesting functional importance. PMID- 15760673 TI - Orexin-B modulates luteinizing hormone and growth hormone secretion from porcine pituitary cells in culture. AB - To test the hypothesis that orexin-B acts directly on the anterior pituitary to regulate LH and growth hormone (GH) secretion, anterior pituitary cells from prepuberal gilts were studied in primary culture. On day 4 of culture, 10(5) cells/well were challenged with 0.1, 10 or 1000 nM GnRH; 10, 100 or 1000 nM [Ala15]-hGRF-(1-29)NH2 or 0.1, 1, 10 or 100 nM, orexin-B individually or in combinations with 0.1 and 1000 nM GnRH or 10 and 1000 nM GRF. Secreted LH and GH were measured at 4 h after treatment. Basal LH and GH secretion (control; n = 6 pigs) was 183 +/- 18 and 108 +/- 4.8 ng/well, respectively. Relative to control at 4 h, all doses of GnRH and GRF increased (P < 0.0001) LH and GH secretion, respectively. All doses of orexin-B increased (P < 0.01) LH secretion, except for the 0.1 nM dose. Basal GH secretion was unaffected by orexin-B. Addition of 1, 10 or 100 nM orexin-B in combinations with 0.1 nM GnRH increased (P < 0.001) LH secretion compared to GnRH alone. Only 0.1 nM (P = 0.06) and 100 nM (P < 0.001) orexin-B in combinations with 1000 nM GnRH increased LH secretion compared to GnRH alone. All doses of orexin-B in combination with 1000 nM GRF suppressed (P < 0.0001) GH secretion compare to GRF alone, while only 0.1 nM orexin-B in combination with 10 nM GRF suppressed (P < 0.01) GH secretion compared to GRF. These results indicate that orexin may directly modulate LH and GH secretion at the level of the pituitary gland. PMID- 15760674 TI - Hyperadrenocorticism in a dog due to ectopic secretion of adrenocorticotropic hormone. AB - Spontaneous hyperadrenocorticism in dogs is known to be the result of excessive secretion of adrenocorticotropic hormone (ACTH) by the pituitary gland or excessive autonomous glucocorticoid secretion by an adrenocortical tumor. Here, we report on an 8-year-old German shepherd dog in which ACTH-dependent hyperadrenocorticism was a result of ectopic ACTH secretion and could be related to an abdominal neuroendocrine tumor. Hyperadrenocorticism was diagnosed on the basis of the history, clinical signs, and elevated urinary corticoid/creatinine ratios (UCCRs; 236 and 350 x 10(-6); reference range < 10 x 10(-6)). The UCCR remained elevated (226 x 10(-6)) after three oral doses of dexamethasone (0.1 mg/kg body weight) at 8-h intervals. Ultrasonography revealed two equivalently enlarged adrenal glands, consistent with adrenocortical hyperplasia. Plasma ACTH concentration was clearly elevated (159 and 188 ng/l; reference range 5-85 ng/l). Computed tomography (CT) revealed that the pituitary was not enlarged. These findings were interpreted as indicating dexamethasone-resistant pituitary dependent hyperadrenocorticism. Transsphenoidal hypophysectomy was performed but within 2 weeks after surgery, there was exacerbation of the clinical signs of hyperadrenocorticism. Plasma ACTH concentration (281 ng/l) and UCCRs (1518 and 2176 x 10(-6)) were even higher than before surgery. Histological examination of the pituitary gland revealed no neoplasia. Stimulation of the pituitary with corticotropin-releasing hormone did not affect plasma ACTH and cortisol concentrations. Treatment with trilostane was started and restored normocorticism. CT of the pituitary fossa, 10 months after hypophysectomy, revealed an empty sella. Hence, it was presumed that there was ectopic secretion of ACTH. CT of the abdomen revealed a mass in the region of the pancreas and a few nodules in the liver. Partial pancreatectomy with adjacent lymph node extirpation was performed and the liver nodules were biopsied. Histological examination revealed a metastasized neuroendocrine tumor. Abdominal surgery was not curative and medical treatment with trilostane was continued. At 18 months after the abdominal surgery, the dog is still in good condition. In conclusion, the combination of (1) severe dexamethasone-resistant hyperadrenocorticism with elevated circulating ACTH levels, (2) definitive demonstration of the absence of pituitary neoplasia, and (3) an abdominal neuroendocrine tumor allowed the diagnosis of ectopic ACTH secretion. PMID- 15760675 TI - Inflammation, genetics, and ischemic heart disease: focus on the major histocompatibility complex (MHC) genes. AB - BACKGROUND: Increasing data suggest that ischemic heart disease (IHD) shares several characteristics with common inflammatory diseases (such as rheumatoid arthritis), in which the pathogenetic role of inflammatory gene polymorphisms is well established. Variants in the genes for the major histocompatibility complex (MHC) molecules on the short arm of chromosome 6 show profound "linkage disequilibrium", leading to the formation of "haplotypes", i.e., frozen blocks of alleles travelling together through generations. DESIGN: We performed a review of published studies linking IHD with gene polymorphisms of the MHC molecules tumor necrosis factor (TNF)-alpha and -beta, the class II DR human leukocyte antigens, heat shock protein 70-1, hemochromatosis related gene, and complement C4. RESULTS: The emerging data are quite conflicting and do not provide definitive evidence for a role of these gene variants in the pathogenesis of IHD; a possible exception is the G252A and polymorphism in the TNF-beta gene (also known as lymphotoxin-alpha) which, in a comprehensive genome-scan linkage analysis of unrelated Japanese, but not in a smaller German population, was linked to myocardial infarction. However, some important biases appear, e.g. different study design and variable linkage disequilibrium among different populations. CONCLUSIONS: Preliminary positive results should encourage future studies to focus on clinical models of IHD with well-codified inflammatory components, using novel methods (such as haplotype analysis) to assess gene polymorphisms and their clinical effect. PMID- 15760676 TI - Contribution of protein kinase A and protein kinase C pathways in ultraviolet B induced IL-8 expression by human keratinocytes. AB - We have previously demonstrated that treatment of the human keratinocyte cell line NCTC 2544 with a UVB dose equivalent to 1h exposure (100 mJ/cm2) results in a significant increase of IL-8 production. In this study, we use specific inhibitors to investigate the role of both PKA- and PKC-mediated pathways in the regulation of UVB-induced IL-8 expression in NCTC 2544 cell line. We show here that the treatment of irradiated human keratinocytes with PKA inhibitors [H89 and PKA inhibitor (PKAi)] induced a significant decrease of IL-8 production at both mRNA and protein levels. However, the regulation of IL-8 production seems to be mediated via a cAMP-independent PKA pathway, since drugs known to enhance cAMP concentrations [PGE2, cholera toxin and dibutyryl cAMP] decrease IL-8 production in irradiated cells by down-regulating NF-kappa B activation in response to UVB radiation. Using PMA (a potent pharmacological activator of PKC) and calphostin C (a specific PKC inhibitor), we demonstrated an up-regulation of IL-8 in NCTC 2544 cells and a down-regulation of the cytokine in UVB-irradiated cells, respectively. We also observed that in our experimental conditions, staurosporine, an inhibitor of both PKC and PMA-stimulated cellular responses, does not involve PKC inhibition in irradiated cells and significantly decreased NF-kappa B activity in response to UVB radiation. Finally, we concluded that a cAMP-independent PKA activation and a PKC-associated pathway are probably involved in the regulation of UVB-induced IL-8 synthesis in human keratinocytes. PMID- 15760677 TI - Contribution of the sympathetic nervous system on the burn-associated impairment of CCL3 production. AB - Previously, we have reported that the susceptibility of burned patients to infectious complications is increased when the production of CC-chemokine ligand 3 (CCL3) is impaired. In this study, the role of the sympathetic nervous system on impaired CCL3 production and antibacterial resistance following burn injuries was investigated. Normal mice were resistant (65% survival) to cecal ligation and puncture (CLP)-induced sepsis, while the same CLP killed 90% of thermally injured mice (TI-mice). However, TI-mice resisted CLP-induced sepsis (60% survival) when they were previously treated with CCL3 or sympathectomized with 6-hydroxydopamine (6-OHDA). Augmentation of host resistance against CLP-induced sepsis by 6-OHDA was abrogated by anti-CCL3 mAb treatment. Norepinephrine (NE) production was increased in circulation of TI-mice, and treatment of TI-mice with 6-OHDA resulted in the inhibition of NE secretion. CCL3 production was impaired in cultures of T cells from TI-mice or normal T cells treated with NE, even when stimulated with anti-CD3 mAb. However, CCL3 was produced by mAb-stimulated T cells from TI-mice previously treated with 6-OHDA. These results indicated that by inhibiting CCL3 production the sympathetic nervous system contributes to the increased susceptibility of TI-mice to sepsis. PMID- 15760678 TI - IL-10 and toll-like receptor-4 polymorphisms and the in vivo and ex vivo response to endotoxin. AB - To determine to what extent lipopolysaccharide-induced IL-10 production capacity is determined by polymorphisms in toll-like receptor-4 (TLR4) and the IL-10 promoter region, we measured in vivo IL-10 and TNF-alpha production in patients undergoing elective cardiopulmonary bypass surgery, a major surgical trauma associated with ischemia-reperfusion injury that triggers an endotoxemia and profound inflammatory response in most patients. Ex vivo the IL-10 and TNF-alpha production was measured in a whole blood stimulation assay, using 3 LPS concentrations. Positive correlations were found between TNF-alpha and IL-10 production ex vivo, upon stimulation with each of the LPS concentrations. Also, the estimated TNF-alpha and IL-10 EC50, and TNF-alpha(max) and IL-10max were positively correlated (r = 0.203; p = 0.023 and r = 0.287; p = 0.001, respectively), indicating that these parameters describing LPS sensitivity and maximal production capacity, respectively, can be estimated by measuring either TNF-alpha or IL-10. Interleukin-10 concentrations in patients experiencing endotoxemia in vivo negatively correlated with the IL-10 levels produced upon stimulation with 1000 ng/mL LPS as well as the estimated IL-10max ex vivo. In vivo, a positive correlation between the TNF-alpha concentration at time-point 2 and the IL-10 concentration at time-point 3 was found, consistent with an important contribution of the magnitude of TNF-alpha release upon the subsequent IL-10 production. Carriers of the IL-10 promoter -1330G, -1082A, -819T, -592A (GATA) haplotype had lower IL-10 production ex vivo upon stimulation with 10 and 100 ng/mL LPS and higher EC50 values (the estimated LPS concentration at which 50% of the maximal IL-10 response is reached) as compared to carriers of the other haplotypes combined, indicating decreased LPS sensitivity ex vivo. These individuals did not differ from the others in interleukin-10 production capacity upon stimulation with a high LPS concentration (i.e., 1000 ng/mL) and the estimated IL-10(max) values, were similar, indicating unimpaired maximal IL-10 production capacity ex vivo. Carriers of the IL-10 promoter AGCC haplotype had lower EC50 values as compared to carriers of the other haplotypes combined, indicating increased LPS sensitivity ex vivo. In accordance with this finding, carriers of the AGCC haplotype had higher circulating IL-10 levels in vivo. The common TLR4 polymorphisms (Asp299Gly and Thr399Ile) were associated with slightly higher IL-10 production capacity ex vivo and in vivo, however, this was not statistically significant. Our results indicate that polymorphisms in the proximal IL-10 promoter region are associated with in vivo and ex vivo LPS sensitivity. The contribution to the inter-individual variation, however, is limited since the variation between individuals in LPS sensitivity and IL-10 production capacity can only partly be attributed to these IL-10 promoter polymorphisms. PMID- 15760679 TI - Involvement of TL1A and DR3 in induction of pro-inflammatory cytokines and matrix metalloproteinase-9 in atherogenesis. AB - TL1A (VEGI/TNFSF15) is the ligand for DR3 (TNFRSF12) and is a newly identified member of the tumor necrosis factor superfamily (TNFSF). Previously, DR3 has been shown to have a role in atherogenesis through stimulation of matrix degrading enzymes including matrix metalloproteinase (MMP)-9. Immunohistochemical staining of human carotid atherosclerotic plaques revealed a high-level expression of TL1A in regions rich in macrophage/foam cells. To investigate the role of TL1A and DR3 in the functioning of macrophage/foam cells in relation to atherogenesis, we have analyzed cellular events mediated by TL1A and DR3 in a human macrophage-like cell line, THP-1. Treatment of THP-1 cells with immobilized anti-DR3 monoclonal antibody in combination with IFN-gamma caused induction of pro-atherogenic cytokines/chemokines such as TNF-alpha, monocyte chemoattractant protein (MCP)-1, and interleukin (IL)-8. Treatment of THP-1 cells with recombinant TL1A in combination with IFN-gamma also caused induction of MMP-9 and IL-8. Furthermore, the expression of DR3 in peripheral blood monocytes was induced after atherogenic stimulation. These data suggest that TL1A and DR3 is involved in atherosclerosis via the induction of pro-inflammatory cytokines/chemokines and decreasing plaque stability by inducing extracellular matrix degrading enzymes. PMID- 15760681 TI - Screening for isolated diastolic dysfunction--a bridge too far? PMID- 15760680 TI - Lack of tumor necrosis factor receptor type 1 inhibits liver fibrosis induced by carbon tetrachloride in mice. AB - Chronic liver injury causes liver regeneration, resulting in fibrosis. The proinflammatory cytokine tumor necrosis factor (TNF) is involved in the pathogenesis of many acute and chronic liver diseases. TNF has pleiotropic functions, but its role in liver fibrosis has not been clarified. Chronic repeated injection of CCl4 induces liver fibrosis in mice. We examined whether signaling through TNF receptors was critical for this process, using mice lacking either TNF receptor (TNFR) type 1 or TNFR type 2 to define the pathophysiologic role of TNFR signals in liver fibrosis. Liver fibrosis caused by chronic CCl4 exposure was TNF-dependent; histological fibrosis was seen in wild-type (WT) and TNFR-2 knockout (KO) mice, but not in TNFR-1 KO mice. Furthermore, a marked reduction in procollagen and TGF-beta synthesis was observed in TNFR-1 KO mice, which also had little detectable NF-kappa B, STAT3, and AP1 binding, and reduced levels of liver interleukin-6 (IL-6) mRNA compared to WT and TNFR-2 KO mice. In conclusion, our results indicate the possibility that NF-kappa B, STAT3, and AP1 binding by signals transduced through TNFR-1 plays an important role in liver fibrosis formation. PMID- 15760682 TI - Tissue motion imaging of the left ventricle. PMID- 15760684 TI - Value of left ventricular filling parameters to predict mortality and functional class in patients with heart disease from the community. AB - Surprisingly few studies described the value of Doppler parameters in patients from the community. AIM: The aim was to determine the prevalence of prolonged IVRT (>0.105s) or a prolonged EDT (>0.280s) with a reduced E/A (<0.5) in heart patients from the community without valvular, systolic or rhythmic dysfunction. The associations of these parameters to all cause mortality and NYHA functional class were examined as well. METHODS: Seventy-two volunteer stable patients with a history of heart disease were identified from general practice. Patients with LVEF below 0.45, valvular abnormalities, atrial fibrillation, and pacemaker were excluded. Routine blood tests, echocardiography, chest X-ray, physical examination and mortality were evaluated. RESULTS: PATIENT FINDINGS: male 33%, mean age of 68 years, hypertension 82%, ischaemic heart disease 43%, and NYHA class I+II+III in 50+39+11%. Abnormal EDT occurred in 4% (95% CI from 0 to 9%), IVRT in 18% (9-27%), E/A in 0%. None had a restrictive pattern. EDT was longer in NYHA III than in NYHA I-II patients (median 0.25 vs. 0.19s, p=0.0006). E/A and IVRT were not associated with NYHA class or mortality. After 7.4 years 16 of 72 patients died. EDT predicted mortality in univariate analysis but not in a multivariate analysis where NYHA class and gender were the only significant predictors. CONCLUSION: Prolonged EDT was weakly associated to NYHA class and mortality while IVRT and E/A were not. Prolonged IVRT was a frequent finding, but a diagnosis of diastolic dysfunction is not supported by mild to moderate abnormal IVRT or E/A. PMID- 15760685 TI - Intracardiac echocardiography: an ideal guiding tool for device closure of interatrial communications. AB - BACKGROUND: This study sought to evaluate safety and radiation exposure when using intracardiac echocardiography (ICE) in comparison to transesophageal echocardiography (TEE) in order to guide transcatheter closure of interatrial communications. METHODS: Eighty patients (44 males, 36 females, mean age 46, SD 13 years) undergoing device closure of atrial septal defect (n=12) or patent foramen ovale (n=68) had the procedure guided by ICE (n=50, group 1) or TEE (n=30, group 2). In group 1, all procedural stages were completely guided by ICE, including imaging of the interatrial communication during balloon sizing, device unfolding and release, and during the final check for adequate positioning. In group 2, exclusive implantation of devices was guided by use of TEE. RESULTS: Especially, the spatial relationship between device and cardiac structures (e.g. the ascending aorta, the interatrial septum and the superior vena cava) was accurately demonstrated in group 1. Image resolution provided by ICE was superior to that of TEE. No severe complications, including any related to ICE, were seen. Fluoroscopy time (FT) and procedure time (PT) were shorter in group 1 than in group 2 (FT: 5.5+/-1.5 min vs. 9.3+/-1.6 min, P<0.0001; PT: 31.9+/-4.6 min vs. 38.8+/-5.8 min, P<0.01). Neither sedation nor anesthesia was required in group 1. CONCLUSIONS: ICE is a safe tool to guide device closure of interatrial communications. For the patient, procedural stress and radiation exposure are negligible. ICE can be considered the guiding tool of choice for device closure, particularly when long or repeated echocardiographic viewing is required. PMID- 15760686 TI - Lateral vs medial mitral annular tissue Doppler in the echocardiographic assessment of diastolic function and filling pressures: which should we use? AB - Tissue Doppler echocardiography (TDE) is used in the assessment of diastolic function, however, it is unclear whether the medial (E' med) or lateral (E' lat) annulus should be used. Our aim was to compare the diagnostic utility of E' med and E' lat. In 232 subjects left ventricular (LV) systolic and diastolic function was assessed via transthoracic echocardiography with TDE measurements obtained from both annuli. LV function was normal in 91 subjects (39%), with diastolic dysfunction found in 141 subjects (61%). TDE velocities decreased with age and progressive diastolic dysfunction for either annulus. E' med recorded significantly lower myocardial velocities than E' lat. Receiver operator curves showed improved area under the curve (AUC) for E' med than E' lat. Furthermore the AUC was significantly improved compared to E/A ratio and deceleration time. For diagnosing diastolic dysfunction, an E' med<11cm/s provided a sensitivity of 78%, specificity of 67% and positive predictive value of 70%. Whilst for diagnosing elevated filling pressures an E/E' medial ratio>8 provided values of 56%, 93% and 91%, respectively. In conclusion, although either annulus can be used, E' med provides better diagnostic utility. PMID- 15760687 TI - Pulmonary venous flow reversal and its relationship to atrial mechanical function in normal subjects--Umea General Population Heart Study. AB - AIMS: Although pulmonary venous flow reversal (Ar) is useful in the evaluation of left ventricular (LV) diastolic function, it is often difficult to study with transthoracic echocardiography (TTE). We determined the relationship between Ar and left atrial (LA) mechanical function and sought to define surrogate measurements for Ar. METHODS AND RESULTS: A total of 130 healthy subjects, mean age 54.3+/-18.3 years, 62 women, were studied and classified into three groups: [young (Y), 25-44 years; n=44], [middle-age (M), 45-64 years; n=43] and [elderly (E), > or =65 years; n=43]. Pulmonary venous flow and LV inflow studies were performed by TTE and LV basal free-wall motion was studied by Doppler tissue imaging (DTI). All images were acquired with a superimposed electrocardiogram. RR interval was similar in all groups while LA dimension and PR interval were increased in Group E vs. Y (P<0.001). LA contraction (A(m)) on DTI, transmitral A wave (A) and Ar were simultaneous and started 84ms after onset of P wave and this interval increased with age (P=0.02). Similarly, the time intervals from the same landmark to peak A(m), A, and Ar were prolonged with age (all, P<0.001). Despite this prolongation, peak A(m) coincided with peak Ar in every age group (r=0.97, P<0.001) and Ar acceleration and deceleration times were consistently equal. CONCLUSION: The timing of A(m) obtained by DTI can be used to accurately estimate corresponding measurements of Ar recorded by TTE in subjects without cardiac disease. PMID- 15760688 TI - Differentiating clinical and echocardiographic characteristics of chordal rupture detected in patients with rheumatic mitral valve disease and floppy mitral valve: impact of the infective endocarditis on chordal rupture. AB - AIMS: We aimed to compare the clinical and echocardiographic correlates of chordal rupture in patients with rheumatic mitral valve disease and floppy mitral valve. METHODS AND RESULTS: The study group comprised of 224 patients who underwent transthoracic and transesophageal echocardiography because of the severe mitral regurgitation. Chordal rupture was detected in 58 (25.9%) out of the 224 patients, in 33 out of the 83 (39.7%) patients with floppy mitral valve, and in 25 out of the 141 (17.7%) patients with rheumatic mitral valve disease. Chordal rupture was more frequently associated with anterior leaflet (80%) in patients with rheumatic mitral valve disease, and posterior leaflet (72.7%) in patients with floppy mitral valve (p<0.05). Univariate correlates of chordal rupture were age, male sex, posterior mitral leaflet thickening and chordal elongation in patients with floppy mitral valve (p<0.05), and chordal shortening (p<0.0001) and infective endocarditis involving mitral anterior leaflet (p<0.05) in rheumatic group. Independent predictors of chordal rupture were age (>50 years), posterior mitral leaflet thickness (> or =0.45cm), and male sex (p<0.05) in patients with floppy mitral valve while infective endocarditis involving mitral anterior leaflet (p<0.05) in patients with rheumatic mitral valve disease. Patients with chordal rupture due to floppy mitral valve had an older age (p<0.0001), a male dominance, longer mitral leaflets and chordae, and a larger mitral annulus circumference (p<0.05) as compared to those with rheumatic chordal rupture. Despite the comparable severity of mitral regurgitation and left atrial diameters between the two groups of chordal rupture (p>0.05), functional class and pulmonary artery systolic pressure were higher, and atrial fibrillation, acute deterioration, infective endocarditis, mitral leaflet rupture and need for mitral valve surgery in the 3 months were more frequent in rheumatic chordal rupture subgroup (p<0.05). CONCLUSION: Chordal rupture seems to be more frequently associated with anterior mitral leaflet in rheumatic mitral valve disease, whereas it was the posterior leaflet in floppy mitral valve. Chordal rupture was related to male sex, older age, posterior leaflet thickening, and chordal elongation in patients with floppy mitral valve. However, infective endocarditis, acute deterioration, and need for early mitral surgery were more frequent in patients with rheumatic chordal rupture. PMID- 15760689 TI - The effects of heart rate on myocardial velocity and atrio-ventricular displacement during exercise with and without beta-blockade: a tissue Doppler echocardiographic study. AB - BACKGROUND: Colour tissue Doppler echocardiography (TDE) allows an objective assessment of regional myocardial function. Peak systolic velocity (PSV) and A-V plane displacement (AVPD) obtained from colour TDE correlate well with changes in cardiac wall motion and can discriminate ischemic areas during stress echocardiography. During exercise, the relationship between PSV and AVPD depends on several factors besides ischemia and should be considered when performing exercise stress echocardiography. AIMS: To investigate the relation between PSV, AVPD and heart rate (HR) during semi-upright exercise with and without beta blockade. SUBJECTS AND METHODS: Twelve healthy men underwent semi-upright exercise stress echocardiography with and without beta-blockade on two separate occasions. Standard echocardiographic projections were used for the stress echocardiography. Grey-scale echocardiographic pictures containing colour TDE information were obtained at rest and during a two-stage exercise test, and the images were analyzed off-line. The PSV and AVPD were measured at four points at the base of the left ventricle at the septum and lateral, inferior and anterior walls. RESULTS: PSV, AVPD and HR gradually increased during exercise. The increases in PSV and AVPD were linearly correlated with the increase in HR. The increases in PSV were significantly lower during exercise with beta-blockade than without beta-blockade (P<0.05). This was not observed in AVPD, as increments were not affected by beta-blockade. CONCLUSION: These data showing a relationship between HR and PSV, and a significantly lower PSV with beta-blockade at a given HR, suggest that PSV is influenced by HR and myocardial contractility, both of which are augmented by physical exercise-induced sympathetic stimulation. PMID- 15760691 TI - Double subepicardial aneurysm after myocardial infarction. AB - The echocardiogram of 68-year old man, admitted with an acute myocardial re infarction revealed the presence, in the middle-apical region of the lateral wall, of two little and contiguous subepicardial aneurysms. PMID- 15760692 TI - Multicentric familial cardiac myxoma. AB - Familial cardiac myxoma is a rare syndrome which constitutes approximately 10% or less of all myxomas. We describe a rare case of LA and LV mass simultaneously in a 35-year-old female presenting to our hospital for evaluation of recurrent cardiac myxoma. Echocardiography revealed both LA and LV mass. Surgery was done and histological findings confirmed the diagnosis of myxoma. PMID- 15760690 TI - Quantitative evaluation of regional endocardial visualisation with second harmonic imaging and contrast left ventricular opacification in heart failure patients. AB - AIMS: Wall motion score index (WMSI) is an important prognostic indicator in heart failure (HF) patients but requires endocardial visualisation. This study evaluated the role tissue harmonic imaging (THI) and contrast opacification (LVO) for improving endocardial visualisation and the determination of WMSI in HF patients. METHODS AND RESULTS: Thirty-one HF patients and 30 controls underwent apical echocardiography with fundamental imaging (FUND), THI and THI with contrast agent (Levovist). Visualisation and motion were graded in the six segments from each of the apical two and four chamber views. Both THI and LVO reduced the percentage of non-visualised segments (FUND 13.6%, THI 5.6%, LVO 2.8%, p=0.01) in the controls, but in HF patients, only THI improved visualisation (% segments not visualised FUND 9.7%, THI 3.5%, LVO 4.8%, p=0.06). The anterior and lateral walls were the least well visualised with FUND, but improved with LVO (anterior p=0.0026, lateral p=0.0003). No improvement was seen in the inferior wall (p=0.30) or septum (p=0.2). WMSI was similar by all methods and negatively correlated with ejection fraction (FUND r=-0.69, THI r=-0.74, LVO r=-0.77, all p<0.001). CONCLUSION: THI improved endocardial visualisation in all subjects and LVO offered additional benefit in the controls, but not in HF patients. Regional endocardial visualisation was inconsistent. Thus, both patient factors and wall segment site need to be considered when using contrast agents for endocardial visualisation. PMID- 15760693 TI - Fatal ventricular arrhythmia as a complication of transesophageal echocardiography. AB - AIM: To report a case of serious ventricular arrhythmia during transesophageal echocardiography. METHODS AND RESULTS: A 58-year-old woman with previous mitral and tricuspid valve replacement and permanent pacemaker implantation suffered from recurrent fever and Staphylococcus aureus bacteremia. Transesophageal echocardiography was performed as part of the assessment for infective endocarditis. During this procedure the patient developed sustained ventricular tachycardia and subsequently ventricular flutter. She was successfully resuscitated. Subsequently the procedure was undertaken under general anesthesia with no complications. CONCLUSION: The increasing use of TEE in a wider spectrum of patients, many of whom are seriously ill, may result in serious side-effects. PMID- 15760694 TI - Diagnosis and treatment of pulmonary tuberculosis at basic health care facilities in rural Vietnam: a survey of knowledge and reported practices among health staff. AB - One fundamental element needed for tuberculosis control programmes to succeed is that health staff should adequately manage persons with both suspected and confirmed tuberculosis. This study assesses the level of knowledge and reported practices regarding tuberculosis among health staff at basic health care facilities in a rural district in Vietnam. A questionnaire consisting of 17 multiple-choice questions, 6 open questions, and five case-studies was completed by 253 health staff. Nearly half of the respondents (47%) answered at least 17 out of 23 questions correctly. The mean knowledge score was 15.59 +/- 3.78 (range 5-23). The mean practice score was 2.03 +/- 1.28 (range 0-5). Health staff knowledge of theoretical aspects was better than knowledge related to patient management. Even staff members who had attended TB training courses had inadequate TB knowledge, particularly in the area of TB control. District hospital staff reported surprisingly lower knowledge scores than community health care staff. Practice competency related to the management of tuberculosis patients during treatment course was low. The importance of targeted education using multiple educational methods needs to be emphasized within the National Tuberculosis Programme. PMID- 15760695 TI - Reducing health disparities through primary care reform: the New Zealand experiment. AB - New Zealand experiences significant health disparities related to both ethnicity and deprivation; the average life expectancy for Maori New Zealanders is 9 years less than for other New Zealanders. The government recently introduced a set of primary care reforms aimed at improving health and reducing disparities by reducing co-payments, moving from fee-for-service to capitation, promoting population health management and developing a not for profit infrastructure with community involvement to deliver primary care. Funding for primary care visits will increase by some 43% over 3 years. This paper reviews policy documents and enrollment and payment data for the first 15 months to assess the likely impact on health disparities. The policy has been successfully introduced; over half the New Zealand population (of four million) enrolled in new Primary Health Organizations within 15 months. Over 400,000 people (half of them in vulnerable groups) gained improved access to primary care subsidies in the first 15 months. The combined effect of new payment rules and the deprived nature of the minority populations was that the average per person payment to PHOs on behalf of Maori and Pacific enrollees was more than 70% greater than the per person amount for other ethnicities for the period. The policy is consistent with the principles of the Alma Alta Declaration. Barriers to successful implementation include the risk of middle class capture of the additional funding; the risk that co-payments are not low enough to improve access for the poor; PHO inexperience; and the small size of many PHOs. Transitional equity and efficiency issues with the use of aggregate population characteristics to target higher subsidies are being ameliorated by the introduction of low cost access based on age. A tension between the twin policy goals of low cost access for all, and very low cost access for the most vulnerable populations is identified as a continuing and unresolved policy issue. PMID- 15760696 TI - Rural perspectives on HIV/AIDS prevention: a comparative study of Thailand and Ghana. AB - The paper compares rural perspectives in Thailand and Ghana on the level of condom acceptance in sexual relations, willingness to test oneself for HIV before and in marriage and sources of information on HIV/AIDS. We also compared the policy approaches to combating HIV/AIDS in both countries. The results indicates that in the villages studied in Thailand, all single men and the majority of the single women were in favour of using condoms in sexual relations. This group also showed a positive attitude to HIV/AIDS test before and in marriage. However, married men in rural Thailand disapproved of the use of condoms with their wives but married women in the sample population were open to the possibility of using condoms. Both married men and women were strongly against HIV/AIDS test in marriage. In contrast to Thailand, most single men in the communities studied in Ghana showed a disapproval to the use of condoms in sexual relations. However, they condoned HIV test before marriage. Married men and women in rural Ghana were against the use of condoms in sexual relations as well as HIV/AIDS test in marriage. In order to mitigate mother-to-child transmission, the Thais applied anti-retroviral drug care for HIV positive pregnant women during pregnancy and after delivery. In Ghana on the other hand, pregnant women were subject to HIV test and counselling. The mode of information acquisition on HIV/AIDS in both countries were through the media, campaigns and village volunteers. Finally, we observed that fighting poverty is a sine qua non for the success of any HIV/AIDS eradication programme. PMID- 15760697 TI - Private and public cross-subsidization: financing Beijing's health-insurance reform. AB - In 1998, the Chinese government proposed a universal health-insurance program for urban employees. However, this reform has been advancing slowly, primarily due to an unpractical financing policy. We surveyed over 2000 families and evaluated the financial impacts of Beijing's reform on public and private enterprises. We found that most state-owned enterprises provided effective health insurance, whereas most private firms did not; overall, 33% of employees had little or no coverage. On average, employees of private firms were healthier and earned more compared to public firms. Because the premium was proportional to income, private firms would pay more for insurance than the predicted health-care expense of their employees. International firms subsidize the most, contributing more than 60% of their insurance premiums to the employees of the public sector. Such an aggressive cross-subsidization policy is difficult to be accepted by private firms. PMID- 15760698 TI - Pharmaceutical policy in Italy: towards a structural change? AB - Italian pharmaceutical policy has recently moved towards a "two lanes" approach, with regulation differing according to a drug's patent status. This study analyses the Italian regulatory framework, focusing on policies related to "off patent" drugs. Three main regulatory innovations have been examined: (i) generics, introduced in Italy for the first time in 1996; (ii) the reference pricing (RP) scheme, under which consumers pay part of the cost of high-priced products; (iii) pharmacists' right of substitution, supported by a regressive margins system. The recent reforms are already producing some worthwhile results, at least in terms of competitive pressure on the (few) substances that run out of patent protection. However, further intervention could be required to achieve long-term sustainability. PMID- 15760699 TI - Does hospital discharge policy influence sick-leave patterns in the case of female breast cancer? AB - The objective was to investigate how differences among hospitals in the shift from in-patient care to day surgery and a reduced hospital length of stay affect the sick-leave period for female patients surgically treated for breast cancer. All women aged 18-64 who were diagnosed with breast cancer in 2000 were selected from the National Cancer Register and combined with data from the sick-leave database of the National Social Insurance Board and the National Hospital Discharge Register (N = 1834). A multi-factorial model was fitted to the data to investigate how differences in hospital care practice affected the length of sick leave. The main output measure was the number of sick-leave days after discharge during the year following surgery. The confounders used included age, type of primary surgical treatment, whether or not lymph node dissection was performed, labour-market status, county, and readmission. Women treated with breast conserving surgery had a 54.7-day (-71.9 < or = CI(95%) < or = -37.5) shorter sick-leave period than those with more invasive surgery. The day-surgery cases had 24.3 (-47.5 < or = CI(95%) < or = -1.1) days shorter sick-leave than those who received overnight care. The effect of the hospital median length of stay (LOS) was U-shaped, suggesting that hospitals with a median LOS that is either short or long are associated with longer sick-leave. In the intermediate range, women treated in hospitals with a median LOS of 2 days had 22 days longer sick leave than those treated in hospitals with a mean LOS of 3 days. This is possibly a sign of sub-optimising. PMID- 15760700 TI - Gender-related differences in general practice preferences: longitudinal evidence from the Netherlands 1982-2001. AB - The proportion of female doctors is increasing. Yet, there is little evidence that demonstrates changing career preferences over a long period, nor do we know the long-term impact of changing male-female ratios in medicine. We explored this within the GP profession. By means of a longitudinal cohort study (postal questionnaires) among all graduated GPs in the Netherlands between 1982 and 2001 we explored trends in career preferences and investigated whether practice preferences of men and women differ over the years. Preference of becoming a GP has significantly decreased among men. GPs prefer group practice more than solo practice. Female doctors were more likely to prefer a small practice and to associate. Men prefer to take over an established practice. Main reasons to abstain from practicing as a GP for men were having found another job or not having found a practice according to their preferences. Main reasons for women were having decided for another job and family life. We conclude that male and female GPs select differently on practice setting. Preferences change through the years but tend to converge. Gender differences are likely due to the circumstance that career choices for men are more influenced by fluctuating labour markets, while female choices are more based on family circumstances. We expect that as more women will become a GP the demand for small group practices will increase. Also, as many female GPs abstain from practicing after having finished a vocational GP training program it will be essential to create work facilities to keep them available for the GP labour market. PMID- 15760701 TI - The decision-making process of health care utilization in Mexico. AB - Using individual-level data from the 2000 Mexican Survey of Satisfaction with Health Services we estimate a two-part negative binomial hurdle model to evaluate the decision-making process of health care utilization in Mexico. We find that there are income-related differences in utilization associated with the first visit to a physician, as well as substantial utilization differences by region, employment, insurance and financial status. There are also income-related differences in the first visit to a specialist but not in the number of days hospitalized. The results suggest that increasing initial access to services via income and insurance coverage and providing financial resources to underserved regions can substantially improve access to care and, ultimately, population health. PMID- 15760702 TI - What lessons can be drawn from tuberculosis (TB) control in China in the 1990s? An analysis from a health system perspective. AB - China has made a significant achievement in tackling the TB epidemic over the last decade, due largely to the implementation of directly-observed treatment strategy (DOT). The cure rate of TB cases reached more than 90% for registered TB patients. However, the case detection rate has, unfortunately, been very low (some 30%). Using available information, this paper identifies four main problems facing TB control in China, these are, low case finding, a substantial proportion of TB patients failing to complete standardised treatment, increased proportion of MDR TB patients, and lack of effective TB control among "floating populations". The paper also analyses the possible causes of these problems associated with socio-economic barriers in care seeking, ineffectiveness of TB services, particularly in poor areas, lack of co-operation between health facilities, and weakness of political and financial commitments of local governments to TB control. The paper ends with the discussion of opportunities and challenges facing TB control and makes recommendations for further actions and research. PMID- 15760703 TI - Findings from a survey on GIS use in the UK National Health Service: organisational challenges and opportunities. AB - The use of geographical information systems (GIS) in a variety of application areas points to an increasing interest in the spatial aspects of health policies. Despite the fact that most public sector organisations in the United Kingdom (UK) have access to such software tools, there has not been a comprehensive review of take-up within the health sector. In this paper, we report on a recently completed mixed-method research project that has examined the current levels of GIS use in the UK National Health Service, and focus our discussion on health authorities (HAs). Couched within the context of previous studies that have been concerned with outlining the types of factors influencing IT implementation in the health sector, we illuminate the importance of behavioural, cultural and organisational factors on the diffusion of GIS in the UK National Health Service. It is noted that very few organisations had a GIS strategy and we contend that if GIS is to play a wider role in addressing issues surrounding 'joined-up' government, more advice and guidance is needed on policies promoting the exchange of geographical data between agencies concerned with tackling health issues. We conclude by drawing attention to the perceived lack of national guidance on GIS matters, more generally, within the UK health sector. PMID- 15760704 TI - Seroepidemiology of varicella-zoster virus infection in a cosmopolitan city (Erzurum) in the eastern Turkey. AB - OBJECTIVE: The aim of the study was to determine VZV seroprevalence under age 30 and to identify the relationship of VZV seroprevalence and several sociodemographic characteristics of the study subjects. The results were presented in order to design a strategy for vaccination against varicella-zoster virus (VZV). MATERIAL AND METHOD: It was planned to include a total of 568 subjects. The sampling method of 30 clusters recommended for field studies was used for selecting subjects of a predetermined number in the rural and urban areas in eastern Turkey. ELISA method was used to examine the blood samples for VZV seropositivity. Age, gender, place of living, educational level, family size and socioeconomic status was investigated in the study subjects. RESULTS: Positive VZV seroprevalence was detected in 78% of 559 subjects. Seroprevalence increased with age. Seroprevalence was 16.67% at the age of 1 year, subsequently increased to 57.58% at the age of 4 years, 70% at the age of 7 years, 92.31% at the age of 10 years and then remained 86.78-96.36% in subjects over the age of 10 years. No association was found between sociodemographic variables studied and prevalence levels of antibodies except for educational level in the 0-14 year group. CONCLUSION: These results suggest that the majority of VZV infections occur during the early childhood; the best option to reduce the circulation of wild type VZV in the population would be the immunization of young children. VZV vaccine should be introduced into the routine childhood vaccination programme in Turkey. PMID- 15760705 TI - Different adhesion and migration properties of human HCV29 non-malignant urothelial and T24 bladder cancer cells: role of glycosylation. AB - In tumour cells, alterations in cellular glycosylation may play a key role in their metastatic behaviour. This study used cell lines having very different behaviour in vivo: HCV29 non-malignant transitional epithelium and T24 bladder transitional cell carcinoma. These differences in behaviour might be due in part to differences in cellular glycosylation patterns. Glycan chain analysis of their glycoproteins was performed with the use of specific lectins. The functional role of carbohydrates was studied by treating these cells with swainsonine, an inhibitor of Golgi alpha-mannosidase II, and in vitro adhesion and migration assays. The adhesion of swainsonine-treated HCV29 and T24 cells was increased on fibronectin and type IV collagen by 1.5- and 2-fold, respectively, whereas adhesion on laminin was virtually unchanged after swainsonine-treatment in HCV29 cells and was increased in T24 cells. Swainsonine treatment reduced the rate of T24 cell migration by 20%. We concluded that beta1-6 branched tri- and tetraantennary complex-type glycans have an important function in adhesion and migration in the studied cell lines. These data support the view that oligosaccharides are involved in several steps of the metastatic process. PMID- 15760707 TI - Repair of oxidative damage of thymine by HeLa whole-cell extracts: simultaneous analysis using a microsupport and comparison with traditional PAGE analysis. AB - In mammalian cells, the base excision repair (BER) pathway allows the remove of small DNA base lesions such as oxidized bases. It is initiated by glycosylases that removed the modified base leaving an abasic site that is subsequently processed by AP endonuclease activities. Measurement of BER activities in cell extracts is time consuming and hazardous when radioactive material is used. We report in this study, the parallelized fluorescent analysis of excision of several oxidation products of thymine by cell extracts. To conduct the study, 5 (hydroxymethyl)uracil, 5-formyluracil, 5-carboxyuracil and formylamine together with uracil and the control thymine, were incorporated into oligonucleotides of identical sequences and paired either with adenine or with guanine containing DNA fragments. The oligonucleotides were fixed by sandwich hybridization in wells of a microplate (OLISA technology). Excision by HeLa extracts of the six different DNA base lesions could be followed simultaneously in the same well. Our results showed that the extent of excision of the lesions was the same on support and in solution using classical PAGE analysis approach with modified (32)P-labeled oligonucleotides. We demonstrated that the simultaneous analysis on support is a successful approach to facilitate high-throughput screening of BER activities present in cell extracts. Moreover, extended study of 5-carboxyuracil revealed that this lesion displays similar biological properties as 5-formyluracil. PMID- 15760708 TI - Structural insights for fatty acid binding in a Lys49-phospholipase A2: crystal structure of myotoxin II from Bothrops moojeni complexed with stearic acid. AB - The crystal structure of dimeric Lys49-phospholipase A2 myotoxin-II from Bothrops moojeni (MjTX-II) co-crystallized with stearic acid (C(18)H(36)O(2)) has been determined at a resolution of 1.8 A. The electron density maps permitted the unambiguous inclusion of six stearic acid molecules in the refinement. Two stearic acid molecules could be located in the substrate-binding cleft of each monomer in positions, which favor the interaction of their carboxyl groups with active site residues. The way of binding of stearic acids to this Lys49-PLA(2)s is analogous to phospholipids and transition state analogues to catalytically active PLA(2)s. Two additional stearic acid molecules were located at the dimer interface region, defining a hitherto unidentified acyl-binding site on the protein surface. The strictly conserved Lys122 for Lys49-PLA(2)s may play a fundamental role for stabilization of legend-protein complex. The comparison of MjTX-II/satiric acid complex with other Lys-PLA(2)s structures whose putative fatty acids were located at their active site is also analysed. Molecular details of the stearic acid/protein interactions provide insights to binding in group I/II PLA(2)s, and to the possible interactions of Lys49-PLA(2)s with target membranes. PMID- 15760706 TI - Identification of galectin-4 isoforms in porcine small intestine. AB - Lactose-binding proteins with molecular masses of 14-, 17-, 18-, 28-, and 34-kDa were identified in extracts from porcine small intestinal mucosa. Amino acid sequence analysis of peptides generated by CNBr cleavage of the 34-kDa protein, the most abundant of these proteins, identified this protein as porcine galectin 4. To determine if a porcine homolog of murine galectin-6 is expressed in small intestine, primers for a reverse transcriptase-polymerase chain reaction (RT-PCR) were developed that amplified across the linker region of galectin-4, which is the region that differs between murine galectins-4 and -6. Using these primers, this RT-PCR approach identified two galectin-4 isoforms that differed in the length of their linker region. The larger isoform, galectin-4.1, is nine amino acids longer in its linker region than the smaller isoform, galectin-4.2. Based on nucleotide sequence similarities, the two isoforms are likely splice variants of galectin-4 pre-mRNA and not products of separate genes like murine galectins-4 and -6. PMID- 15760709 TI - Human endo-alpha1,2-mannosidase is a Golgi-resident type II membrane protein. AB - The cDNA for human endo-alpha1,2-mannosidase was reconstructed using two independent EST-clones and its properties characterized. The 2837 bp cDNA construct contained a 1389 bp open reading frame (ORF) encoding for 462 amino acids and an approximately 53.6 kDa protein, respectively. Hydrophobicity analysis of this amino acid sequence, as well as proteolytic degradation studies, indicate that the enzyme is a type II protein, anchored in the membrane via a 19 amino-acid long apolar sequence close to the N-terminus. Human endo-alpha1,2 mannosidase displays a high degree of sequence identity with the catalytic domain of the homologous rat liver endo-enzyme, but differs substantially in the N terminal peptide region, which includes the transmembrane domain. No sequence similarity exists with other processing alpha-glycosidases. Based on sequence information provided by the 2837 bp construct, the cDNA consisting of the complete 1389 bp ORF was amplified by RT-PCR using human fibroblast RNA. Incubation of E. coli lysates with this cDNA, previously modified for boost translation by codon optimization, resulted in the synthesis of an approximately 52 kDa protein which degraded [(14)C]Glc(3)-Man(9)-GlcNAc(2) efficiently, indicating that the catalytic domain of the enzyme folds correctly under cell free conditions. Transfection of the endo-alpha1,2-mannosidase wild-type cDNA into COS 1 cells resulted in a moderate (approximately 1.5-fold) but reproducible increase of activity compared with control cells, whereas >18-fold increase in activity was measured after expression of a chimera containing green-fluorescent protein (GFP) attached to the N-terminus of the endo-alpha1,2-mannosidase polypeptide. This, together with the observation that GFP-endo-alpha1,2 mannosidase is expressed as a Golgi-resident type II protein, points to enzyme specific parameters directing folding and membrane anchoring, as well as Golgi targeting, not being affected by fusion of GFP to the endo-alpha1,2-mannosidase N terminus. PMID- 15760710 TI - Membrane viscosity is a major modulating factor of the enterocin CRL35 activity. AB - Enterocin CRL35 activity is deeply influenced by the membrane viscosity as could be demonstrated performing determinations of the minimal inhibitory concentrations (MIC) at different temperatures and analyzing the membrane viscosity in these cells as well as in resistant bacteria. In all the cases, bacteriocin activity was linked to higher levels of viscosity. This finding was confirmed studying the interaction of enterocin CRL35 with liposomes composed of dimyristoyl phosphatidylcholine: dimyristoyl phosphatidylglycerol (9:1) in both gel and liquid-crystalline phases. It could be establish, from peptide insertion analysis following the tryptophan fluorescence and microviscosity experiments that this peptide is able to interact more efficiently with membranes having a more structured hydrophobic core. PMID- 15760711 TI - Kinetic properties of glucose-6-phosphate dehydrogenase from lamb kidney cortex. AB - Glucose-6-phosphate dehydrogenase is the key regulatory enzyme of the pentose phosphate pathway and one of the products of this enzyme; NADPH has a critical role in the defence system against the free radicals. In this study, glucose-6 phosphate dehydrogenase from lamb kidney cortex kinetic properties is examined. The purification procedure is composed of two steps after ultracentrifugation for rapid and easy purification: 2', 5'-ADP Sepharose 4B affinity and DEAE Sepharose Fast Flow anion exchange chromatography. Previously, we used this procedure for the purification of glucose-6-phosphate dehydrogenase from bovine lens. The double reciprocal plots and product inhibition studies showed that the enzyme obeys 'Ordered Bi Bi' mechanism: K(m NADP+)K(m G-6-P) and K(i G-6-P) (dissociation constant of the enzyme--G-6-P complex) were found to be 0.018 +/- 0.002, 0.039 +/- 0.006 and 0.029 +/- 0.005 mM, respectively, by using nonlinear regression analysis. The enzyme was stable at 4 degrees C for a week. PMID- 15760712 TI - YidC-dependent translocation of green fluorescence protein fused to the FliP cleavable signal peptide. AB - Escherichia coli FliP is a rare bacterial polytopic membrane protein synthesized with a cleavable, highly hydrophobic signal peptide. More hydrophilic Tat dependent or Sec-dependent signal peptide is functionally capable of substituting for the FliP signal peptide, but a signal anchor of inner membrane protein fails to do so. To assess the intrinsic characteristics of the FliP signal peptide in mediating protein translocation, we fused it to green fluorescence protein and observed that the translocation of the chimera (FliPss-GFP) was dependent of Ffh, SecA, SecY and SecD. In addition, we showed for the first time the involvement of YidC in protein translocation across the inner membrane. PMID- 15760714 TI - Functional and structural analysis of catalase oxidized by singlet oxygen. AB - Purified catalase-1 (CAT-1) from Neurospora crassa asexual spores is oxidized by singlet oxygen giving rise to active enzyme forms with different electrophoretic mobility. These enzyme forms are detected in vivo under stress conditions and during development at the start of the asexual morphogenetic transitions. CAT-1 heme b is oxidized to heme d by singlet oxygen. Here, we describe functional and structural comparisons of the non-oxidized enzyme with the fully oxidized one. Using a broad H(2)O(2) concentration range (0.01-3.0 M), non-hyperbolic saturation kinetics was found in both enzymes, indicating that kinetic complexity does not arise from heme oxidation. The kinetics was consistent with the existence of two kinds of active sites differing more than 10-times in substrate affinity. Positive cooperativity for one or both of the saturation curves is possible. Kinetic constants obtained at 22 degrees C varied slightly and apparent activation energies for the reaction of both components are not significantly different. Protein fluorescence and circular dicroism of the two enzymes were nearly identical, indicating no gross conformational change with oxidation. Increased sensitivity to inhibition by cyanide indicated a local change at the active site in the oxidized catalase. Oxidized catalase was less resistant to high temperatures, high guanidinium ion concentration, and digestion with subtilisin. It was also less stable than the non-oxidized enzyme at an acid pH. The overall data show that the oxidized enzyme is structurally different from the non-oxidized one, although it conserves most of the remarkable stability and catalytic efficiency of the non-oxidized enzyme. Because the enzyme in the cell can be oxidized under physiological conditions, preservation of functional and structural properties of catalase could have been selected through evolution to assure an active enzyme under oxidative stress conditions. PMID- 15760713 TI - Large scale in vivo synthesis of globotriose and globotetraose by high cell density culture of metabolically engineered Escherichia coli. AB - Large amounts of globotriose (Galalpha-4Galbeta-4Glc) are shown to be produced by the high cell density culture of an Escherichia coli strain over-expressing the Neisseria meningitidis lgtC gene for alpha-1,4-Gal transferase. The strain which was devoid of both alpha and beta galactosidase activity was fed with glycerol as the energy and carbon source and with lactose as precursor for globotriose synthesis. After complete exhaustion of lactose, globotriose could serve as an alternative acceptor for LgtC and the formation of a series of polygalactosylated compounds was observed. The system was extended to the synthesis of globotetraose (GalNAcbeta-3Galalpha-4Galbeta-4Glc) by overexpressing two additional genes: lgtD from Haemophilus influenzae Rd which encodes a beta-1,3-GalNAc transferase and wbpP from Pseudomonas aeruginosa which encodes a UDP-GalNAc C4 epimerase. Globotetraose could also be produced from exogenous globotriose which was shown to be actively taken up by the cells. PMID- 15760715 TI - Non-catalytic domains of subunit A negatively regulate the activity of calcineurin. AB - Calcineurin is composed of a catalytic subunit A (CNA) and a regulatory subunit B (CNB). In addition to the catalytic core, CNA further contains three non catalytic domains--CNB binding domain (BBH), calmodulin binding domain (CBD), and autoinhibitory domain (AI). To investigate the effect of these three domains on the activity of CNA, we have constructed domain deletion mutants CNAa (catalytic domain only), CNAac (CNAa and CBD), and CNAaci (CNAa, CBD and AI). By using p nitrophenylphosphate and (32)P-labeled R(II) peptide as substrates, we have systematically examined the phosphatase activities, kinetics, and regulatory effects of Mn(2+)/Ni(2+) and Mg(2+). The results show that the catalytic core has the highest activity and the order of activity of the remaining constructs is CNAac>CNAaci>CNA. Sequential removal of the non-catalytic domains corresponds to concurrent increases of the phosphatase activity assayed under several conditions. This observation clearly demonstrates that non-catalytic domains negatively regulate the enzyme activity and act as intra-molecular inhibitors, possibly through restraining the conformation elasticity of the catalytic core required for optimal catalysis or interfering with substrate access. The sequential domain deletion favors activation of the enzyme by Mn(2+)/Ni(2+) but not by Mg(2+) (except for CNAa), suggesting that enzyme activation by Mn(2+)/Ni(2+) is mainly mediated via the catalytic domain, whereas activation by Mg(2+) is via both the catalytic core and non-catalytic domains. PMID- 15760716 TI - Synthesis and anti-elastase properties of 6-amino-2-phenyl-4H-3,1-benzoxazin-4 one aminoacyl and dipeptidyl derivatives. AB - A series of 6-amino-2-phenyl-4H-3,1-benzoxazin-4-one aminoacyl and dipeptidyl derivatives, in which aminoacids and dipeptides are linked to the benzoxazinone moiety via an amide bond, were synthesized and tested in vitro for their inhibitory activity towards human leukocyte elastase (HLE). When compared to their values without inhibitors, the residual enzymatic activities decrease with time, indicating a time-dependent inhibition. The most potent inhibitions were obtained when Z-Arg-(Pmc), Z-Val-Phe, Z-Ala-Val or Z-Val-Ala are linked to the 6 amino group. Twenty-five new compounds were synthesized. PMID- 15760717 TI - The PatB protein of Bacillus subtilis is a C-S-lyase. AB - The PatB protein of Bacillus subtilis had both cystathionine beta-lyase and cysteine desulfhydrase activities in vitro. The apparent K(m) value of the PatB protein for cystathionine was threefold higher than that of the MetC protein, the previously characterized cystathionine beta-lyase of B. subtilis. In the presence of cystathionine as sole sulfur source, the patB gene present on a multicopy plasmid restored the growth of a metC mutant. In addition, the patB metC double mutant was unable to grow in the presence of sulfate or cystine while the patB or metC single mutants grew similarly to the wild-type strains in the presence of the same sulfur sources. In a metC mutant, the PatB protein can replace the MetC enzyme in the methionine biosynthetic pathway. PMID- 15760718 TI - Strategic approaches for overcoming peptide and protein instability within biodegradable nano- and microparticles. AB - This paper reviews the major factors that are closely involved in peptide and protein degradation during the preparation of biodegradable nano- and microparticles. The various means usually employed for overcoming these obstacles are described, in order to bring to the fore the strategies for protein stabilization. Both processing and formulation parameters can be modified and are distinctly considered from a strategic point of view. We describe how partial or full protein stability retention within the carriers and during drug release might be achieved by individual or combined optimized strategies. Additionally, problems commonly encountered during protein quantification, stability determination and release are briefly reviewed. Artefacts that might occur during sampling and analytical procedures and which might hinder critical interpretation of results are discussed. PMID- 15760719 TI - The therapeutic antibodies market to 2008. AB - The therapeutic biologics market is currently dominated by recombinant protein products. However, many of these products are mature, and growth of the biologics market will increasingly rely on the expansion of the therapeutic monoclonal antibody sector. Successive technology waves have driven the growth of the monoclonal antibody sector, which is currently dominated by chimeric antibodies. Chimeric products, led by Remicade and Rituxan, will continue to drive market share through to 2008. However, over the forecast period, humanized and fully human monoclonal antibodies, together with technologies such as Fabs and conjugated antibodies, will play an increasingly important role, driving monoclonal antibody market growth at a forecast compound annual growth rate of 20.9%, to reach $16.7 billion by 2008. In terms of therapeutic focus, the monoclonal antibody market is heavily focused on oncology and arthritis, immune and inflammatory disorders, and products within these therapeutic areas are set to continue to be the key growth drivers over the forecast period. Underlying the growth of the market is the evolution of the monoclonal antibody company business model, set to transition towards the highly successful innovator model. PMID- 15760720 TI - Manufacturing, regulatory and commercial challenges of biopharmaceuticals production: a Finnish perspective. AB - Biopharmaceuticals product development is a broad and multidisciplinary field. Science and technology are combined with new manufacturing, regulatory and commercial challenges. However, although there is ample literature on the molecular biology and biochemistry of products, the implementation of processes from test tube to commercial scale has not received similar attention. Consequently, the present study aims to highlight, from practical point of view, some of the key issues involved with manufacturing technologies of biopharmaceuticals at a commercial scale. Regulatory requirements and investments are also addressed based on the practical experiences of start-up and small companies. Finland is used as a case-example of such companies as this is a EU member state with strong technological growth and rapidly increasing number of biotech companies. PMID- 15760722 TI - Albumin-protamine-oligonucleotide nanoparticles as a new antisense delivery system. Part 1: physicochemical characterization. AB - In this paper, a ternary system of albumin-protamine-oligonucleotide nanoparticles (AlPrO-NP) recently developed by Vogel et al. [V. Vogel, D. Lochmann, J. Weyermann, G. Mayer, C. Tziatzios, J.A. van den Broek, W. Haase, D. Wouters, U.S. Schubert, J. Kreuter, A. Zimmer, D. Schubert, Oligonucleotide protamine-albumin nanoparticles: preparation, physical properties and intracellular processing, J. Controlled Rel. (in press)] which could serve as a potential drug delivery system for antisense oligonucleotides. Former studies of binary protamine-oligonucleotide nanoparticles showed two main disadvantages: (i) aggregation of the particles within a few minutes in the presence of salt; (ii) low intracellular dissociation between protamine and oligonucleotide, especially phosphorothioates. To overcome these problems, human serum albumin (HSA) as a non toxic, biodegradable macromolecule was introduced as protective colloid. The assembly process of AlPrO-NP was investigated by small angle X-ray scattering (SAXS), fluorescence correlation spectroscopy (FCS), photon correlation spectroscopy (PCS) measurements and scanning electron microscopy (SEM). 'Initial complexes' of HSA and protamine sulphate with a mean hydrodynamic diameter (dh) of about 10-14 nm were found. After adding oligonucleotides (unmodified, phosphorothioate DNA and small interfering RNA), nanoparticles (NPs) were assembled in water and in isotonic media with a dh in a range of 230-320 nm for most preparations. The chemical composition of the particles was investigated by high performance liquid chromatography and fluorescence spectrometry. The whole amount of oligonucleotides (30 microg) was entrapped into the particles at a 1:2 mass ratio (oligonucleotide/protamine). Approximately 7-10% (w/w) of the HSA was bound to the particles. The surface charge of the particles ranged from about +12 to -60 mV depending on the protamine concentration and the ionic conditions. The size and the molecular weight of the components, initial complexes and two model NP preparations were calculated from FCS data. These data verified the PCS, SEM and SAXS measurements. PMID- 15760721 TI - Induction and analysis of aggregates in a liquid IgG1-antibody formulation. AB - The objective of this study was to compare different agitation stress methods (stirring in Reacti Vials versus horizontal shaking) in their effect on protein destabilization, to assess several analytical techniques (light obscuration, turbidimetric and light scattering analysis) for detection of aggregates of various sizes and to evaluate the protecting effect of polysorbate 80 on protein aggregation. A monoclonal IgG1 antibody was used as model protein. Both mechanical stress methods can provoke aggregate formation. The method of stirring induces particles in the range of 10-25 microm comparable to shaking stress. However, stirred samples show a much higher absorbance and reveal a second particle species in DLS analysis, suggesting that stirring stress induces a higher amount of smaller protein aggregates. Addition of polysorbate 80 protects the antibody against aggregation. Only in stirred samples a slight increase in sub-visible particles and turbidity was noted. However, a greater extent of aggregation products was detected by DLS as compared to surfactant-free formulations. Thus, polysorbate 80 appears to stabilise small aggregates and prevents further proceeding of the aggregation process. The induction of aggregates by stirring stress in Reacti Vials analysed by absorbance measurement seems to be a good combination for high-throughput formulation studies. PMID- 15760723 TI - Albumin-protamine-oligonucleotide-nanoparticles as a new antisense delivery system. Part 2: cellular uptake and effect. AB - Antisense oligonucleotides have been used as a specific tool to inhibit the expression of disease associated genes for many years. Unfortunately, oligonucleotides are polyanionic macromolecules which have a weak permeability through biological membranes and are rapidly degraded by nucleases. The purpose of this work is to characterise a new drug delivery system developed by [V. Vogel, D.Lochmann, J. Weyermann, G. Mayer, C. Tziatios, J.A. van der Brock, W. Haase, D. Wouters, U.S. Schubert, J. Kreuter, A. Zimmer, D. Schubert, Oligonucleotide-protamine-albumin nanoparticles preparation, physical properties and intracellular processing, J. Controlled Rel. (in press)] which allows an increased cellular uptake and an intracellular dissociation of the oligonucleotides. The new system based on nanoparticles (NPs) consists of human serum albumin, protamine sulphate and antisense-oligonucleotides (AlPrO). We tested these new nanoparticles on mouse fibroblasts which were stably transfected with a N-methyl-D-aspartate (NMDA) receptor (NR). This cell line enabled us to perform in vitro studies of cellular uptake, intracellular dissociation and effect of the antisense-oligonucleotide in a simple excitotoxicity model. We compared our findings with free oligonucleotides and a commercial available liposomal preparation (DOTAP). We found a 12-fold increased cellular uptake of oligonucleotides in comparison to free oligonucleotides while 100% of the cells were transfected. The AlPrO-NPs showed very low cytotoxic side effects during a 24 h application. We saw an antisense effect of about 35% in a functional assay as well as on the protein level (western blot). The results of the cell penetration and the antisense assay demonstrated that AlPrO nanoparticles are promising carriers for oligonucleotide administration. PMID- 15760724 TI - Development and in vitro evaluation of a liposome based implant formulation for the decapeptide cetrorelix. AB - Semisolid phospholipid dispersions of vesicular morphology, so-called vesicular phospholipid gels (VPGs), were prepared by high-pressure homogenisation and tested in vitro for their suitability as implantable sustained release system for the decapeptide cetrorelix, a potent LH-RH antagonist. The VPGs contained 300-500 mg/g egg phosphatidylcholine (E80) and 0.5-10 mg/g cetrorelix acetate (CXA). The in vitro release experiments showed a wide variability of the system in release, ranging from complete release within less than 24 h (0.5mg/g CXA; 400 mg/g E80) to a predicted 80% sustained release over 3 months (8.6 mg/g CXA; 280 mg/g E80). Erosion of the phospholipid matrix, i.e. release of phospholipid vesicles was found to be the main release mechanism, following zero order or first order kinetics depending on the composition of the VPG. CXA-concentration dependent drug-drug or drug-lipid interactions are assumed to be responsible for the change in release kinetics and the decrease of CXA release at high concentrations of the peptide. Multivariate analysis revealed that both lipid concentration and peptide concentration and also the interactions between the two factors are significant factors for the release rate of the peptide. In summary: based on the presented in vitro release data sustained release of therapeutically relevant CXA doses over up to 6 weeks appears feasible. VPGs are thus considered as a promising new approach for the sustained release of peptide hormones. PMID- 15760725 TI - Baclofen-loaded microspheres: preparation and efficacy testing in a new rabbit model. AB - Intrathecal baclofen is the reference treatment for severe spasticity. This drug has to be injected chronically in the intrathecal space by implanted pumps which are very expensive, uncomfortable and sometimes lead to side effects. Previous work has been performed by our group to assess the feasibility of encapsulating baclofen into poly(lactide-co-glycolide) (PLGA) microspheres and injecting these preparations in the intrathecal space of rabbits. The aims of the present study were to improve the encapsulation process for industrial application (scale-up), and to set up an animal model to assess the duration of effect of the new formulations. Modifications included the replacement of methylene chloride by a less toxic solvent, ethyl acetate, and the use of high molecular weight polymers to extend the release rate of the drug. The temperature and organic solvent extraction rate were fully controlled during the whole manufacturing process. All these modifications resulted in high quality microsphere batches with a CV inferior to 5% for encapsulation efficiency and drug loading. Encapsulation efficiency and release patterns were dependent on the drug payload and the polymer used. A formulation displaying a sustained release of baclofen over 174 days and a moderate burst effect of 16% in the first day in vitro was evaluated in a new reliable model of baclofen activity based on electrophysiological measurement of H-reflex in the rabbit. The activity of a very low dose of baclofen microspheres in vivo was sustained over 35 days. Furthermore, the preparation was well tolerated. These newly developed preparations are a very promising approach for enhancing the efficacy and comfort of patients undergoing spasticity treatment. PMID- 15760726 TI - Stability of different formulations and ion pairs of hypericin. AB - Hypericin, solubilized in an instillation fluid consisting of an aqueous buffer supplemented with 1% plasma proteins, is currently used as a clinical diagnostic tool for the detection of superficial TCC (transitional cell carcinoma) tumors. However, the development of a sterile and stable hypericin stock formulation, excluding the presence of plasma constituents, would be an important factor in a more general clinical application of the method. Therefore, we investigated the stability of several heat sterilized hypericin formulations and ion pairs. Besides sodium hypericinate (in distilled water, in phosphate buffer, in polyethyleneglycol (PEG) 400), several other hypericinate salts (potassium, lysine, TRIS or hexylamine) were investigated. As to that, the physical appearance of different hypericin concentrates stored at 4 and 37 degrees C was investigated. Besides, after dilution into cell culture medium, the ability of hypericin remaining to accumulate in tumor cells and demonstrating photocytotoxic effects upon light irradiation was assessed. These findings suggest that PEG 400 is an excellent hypericin formulation, since it maintained the stability of the compound for at least 120 d when stored at either 4 or 37 degrees C. PEG 400 therefore is a suitable vehicle for the storage of hypericin prior to preparation of the bladder instillation solution. PMID- 15760727 TI - Determination of hypericin in human plasma by high-performance liquid chromatography after intravesical administration in patients with transitional cell carcinoma of the bladder. AB - In the present study, the systemic absorption of hypericin was investigated after intravesical instillation of the compound in nine patients with superficial transitional cell carcinoma (TCC) bladder tumors. Hypericin (8 microM) was instilled in the bladder for 2-3 h before photodynamic diagnosis of bladder tumors. Blood was then collected from a peripheral vein 1 h after termination of the instillation. Solid phase extraction with ammonium acetate buffer and methanol was used to extract hypericin from the plasma. A reversed-phase high performance liquid chromatographic method with fluorescence detection was used to identify and quantify hypericin in the extracts from plasma samples. Analysis of standard plasma samples, which were spiked with known amounts of hypericin, indicated that the pH of the buffer was a determining factor in the extraction yield. The results obtained using ammonium buffer (pH 3.5) and methanol showed the mean extraction recovery of hypericin to be 64% (RSD=12%, n=6). The limits of detection and quantification were 6 and 20 nM, respectively. Extraction and analysis of the plasma of patients after intravesical administration showed hypericin concentrations below the detection limit (<6 nM). In addition, photodynamic treatment of in vitro cultured HeLa cells incubated with 1-100 nM hypericin concentrations showed that lower concentrations (1-20 nM) of hypericin do not induce significant photocytotoxic effects. Taken together, these results imply that photosensitization or other systemic side effects in patients are not to be expected after photodynamic diagnosis of TCC bladder tumors with hypericin. PMID- 15760728 TI - A comparison between povidone-ethylcellulose and povidone-eudragit transdermal dexamethasone matrix patches based on in vitro skin permeation. AB - The present study was designed to develop a suitable matrix type transdermal drug delivery system (TDDS) of dexamethasone using blends of two different polymeric combinations, povidone (PVP) and ethylcellulose (EC) and Eudragit with PVP. Physical studies including moisture content, moisture uptake, flatness to study the stability of the formulations and in vitro dissolution of the experimental formulations were performed to determine the amount of dexamethasone present in the patches were performed and scanning electron microscopy (SEM) photographs of the prepared TDDS were taken to see the drug distribution pattern. Drug-excipient interaction studies were carried out using Fourier transform infrared (FTIR) spectroscopic technique. In vitro skin permeation study was conducted in a modified Franz's diffusion cell. All the formulations were found to be suitable for formulating in terms of physicochemical characteristics and there was no significant interaction noticed between the drug and polymers used. In vitro dissolution studies showed that the drug distribution in the matrix was homogeneous and the SEM photographs further demonstrated this. The formulations of PVP:EC provided slower and more sustained release of drug than the PVP:Eudragit formulations during skin permeation studies and the formulation PVP:EC (1:5) was found to provide the slowest release of drug. Based on the above observations, it can be reasonably concluded that PVP-EC polymers are better suited than PVP-Eudragit polymers for the development of TDDS of dexamethasone. PMID- 15760729 TI - Proniosomes as a drug carrier for transdermal delivery of ketorolac. AB - Niosomes are nonionic surfactant vesicles that have potential applications in the delivery of hydrophobic and hydrophilic drugs. Permeation of a potent nonsteroidal anti-inflammatory, ketorolac, across excised rabbit skin from various proniosome gel formulations was investigated using Franz diffusion cells. Each of the prepared proniosomes significantly improved drug permeation and reduced the lag time (P<0.05). Proniosomes prepared with Span 60 provided a higher ketorolac flux across the skin than did those prepared with Tween 20 (7- and 4-fold the control, respectively). A change in the cholesterol content did not affect the efficiency of the proniosomes, and the reduction in the lecithin content did not significantly decrease the flux (P>0.05). The encapsulation efficiency and size of niosomal vesicles formed by proniosome hydration were also characterized by specific high performance liquid chromatography method and scanning electron microscopy. Each of the prepared niosomes achieved about 99% drug encapsulation. Vesicle size was markedly dependent on the composition of the proniosomal formulations. Proniosomes may be a promising carrier for ketorolac and other drugs, especially due to their simple production and facile up. PMID- 15760730 TI - Development and characterization of PLGA nanospheres and nanocapsules containing xanthone and 3-methoxyxanthone. AB - The aim of the present work was to develop and characterize two different nanosystems, nanospheres and nanocapsules, containing either xanthone (XAN) or 3 methoxyxanthone (3-MeOXAN), with the final goal of improving the delivery of these poorly water-soluble compounds. The xanthones-loaded nanospheres (nanomatrix systems) and nanocapsules (nanoreservoir systems), made of poly(DL lactide-co-glycolide) (PLGA), were prepared by the solvent displacement technique. The following characteristics of nanoparticle formulations were determined: particle size and morphology, zeta potential, incorporation efficiency, thermal behaviour, in vitro release profiles and physical stability at 4 degrees C. The nanospheres had a mean diameter <170 nm, a narrow size distribution (polydispersity index <0.1), and a negative surface charge (zeta potential <-36 mV). Their incorporation efficiencies were 33% for XAN and 42% for 3-MeOXAN. The presence of the xanthones did not affect the nanospheres size and zeta potential. DSC studies indicated that XAN and 3-MeOXAN were dispersed at a molecular level within the polymeric nanomatrix. Nanocapsules were also nanometric (mean size <300 nm) and exhibited a negative charge (zeta potential < 36 mV). Their incorporation efficiency values (>77%) were higher than those corresponding to nanospheres for both xanthones. The release of 3-MeOXAN from nanocapsules was similar to that observed for the correspondent nanoemulsion, indicating that drug release is mainly governed by its partition between the oil core and the external aqueous medium. In contrast, the release of XAN from nanocapsules was significantly slower than from the nanoemulsion, a behaviour that suggests an interaction of the drug with the polymer. Nanocapsule formulations exhibited good physical stability at 4 degrees C during a 4-month period for XAN and during a 3-month period for 3-MeOXAN. PMID- 15760732 TI - Starch-dextrin mixtures as base excipients for extrusion-spheronization pellets. AB - Extrusion-spheronization pellets are generally produced with microcrystalline cellulose (MCC) as the principal excipient, giving rise to particles of very high quality. A number of alternative excipients have been proposed and evaluated, mostly other cellulose derivatives (e.g. different grades of Avicel), or mixtures of MCCs and other excipients. In the present study, we evaluated the possible use of starch+agglutinant mixtures as principal excipients for extrusion spheronization pellets, with the aim of producing pellets with more suitable properties for certain types of release. We first characterized the different excipients in terms of morphometry and basic physical properties. Subsequently, torque-rheometry was used to characterize the rheology of wetted masses of the different excipients and excipient mixtures, with the aim of determining optimal amount of wetting agent (water). We also evaluated the water absorption and water retention capacities of each excipient. In view of the results obtained, we produced pellets with the different starch+agglutinant mixtures (but without drug), and used image analysis to characterize pellet morphology. Our results show that some of the mixtures-notably starch (corn starch or wheat starch)+20% white dextrin-gave high-quality pellets with good size and shape distributions. In addition, the properties of the different materials tested suggest that it may be possible to obtain pellets with very different properties. PMID- 15760731 TI - Rheological characterization and turbidity of riboflavin-photosensitized changes in alginate/GDL systems. AB - Riboflavin (RF) in combination with light, in the wavelength range of 310-800 nm, is used to induce degradation of alginic acid gels. Light irradiation of alginate solutions in the presence of RF under aerobic conditions causes scission of the polymer chains. In the development process of a new drug delivery system, RF photosensitized degradation of alginic acid gels is studied by monitoring changes in the turbidity and rheological parameters of alginate/glucono-delta-lactone (GDL) systems with different concentrations of GDL. Addition of GDL induces gel formation of the samples by gradually lowering the pH-value of the system. The turbidity is measured and the cloud point determined. The turbidity starts to increase after shorter times with enhanced concentration of GDL. Enhanced viscoelasticity is detected with increasing GDL concentration in the post-gel regime, but small differences are detected at the gel point. The incipient gel is 'soft' and has an open structure independent on the GDL concentration. In the post-gel regime solid-like behavior is observed, this is more distinct for the systems with high GDL concentrations. The effect of photosensitized RF on alginate/GDL systems decreases with increasing amount of GDL in the system. The same trend is detected whether the systems are irradiated in the pre-gel or in the post-gel regime. PMID- 15760733 TI - Spectrofluorimetric study of eflucimibe-gamma-cyclodextrin inclusion complex. AB - Eflucimibe, a novel and highly potent acyl-coenzyme A cholesterol O-acyl transferase (ACAT) inhibitor, is sparingly soluble in aqueous media and exhibits a very weak natural fluorescence. However, when increasing concentrations of gamma-cyclodextrin (gamma-CD) are added, an increase in the fluorescence signal is observed, attesting the formation of a non-covalent inclusion complex between eflucimibe and the gamma-CD. In this work, the stoichiometry of the complex and the corresponding association constant have been determined from fluorescence data by Benesi-Hildebrand's method (double reciprocal plots). As a result, a 1:1 stoichiometric ratio and a 20 M(-1) formation constant were obtained. This apparent formation constant was determined in water containing 10% methanol, which was needed to improve 'aqueous' solubility of the drug in a CD-free medium. Owing to the extreme hydrophobicity of eflucimibe, these results provide valuable information for pharmaceutical formulation studies. PMID- 15760735 TI - Water sorption-desorption behaviour of methyl methacrylate-starch copolymers: effect of hydrophobic graft and drying method. AB - A new family of graft copolymers combining hydrophilic and hydrophobic components have recently been proposed as direct compression excipients. Copolymers were synthetised by free radical copolymerisation of starch derivatives with methyl methacrylate (MMA) and were alternatively dried by oven or freeze-drying techniques. The aim of this study was to investigate the water vapour sorption desorption behaviour of these copolymers, focusing on the influence of variables such as the hydrophobic component and the drying process. Moisture sorption and desorption isotherms were measured at 25 degrees C and analysed according to GAB and Young-Nelson equations, which distinguish between different physical forms of moisture distribution. The Young-Nelson model gave the best fit to the experimental data. The results obtained showed that the presence of the acrylic component modified not only the total hygroscopicity of the copolymers, as compared with the original starch derivatives, but also the water distribution in the solid, which might have an important role in the effect of moisture content on copolymer characteristics. The main water-sorption mechanism seemed to be absorption into the copolymer particles structure, in agreement with their starching nature. In terms of water sorption-desorption characteristics, no marked differences were found between the two drying methods used. PMID- 15760734 TI - Effects of roller compaction settings on the preparation of bioadhesive granules and ocular minitablets. AB - An experimental factorial design was employed to evaluate bioadhesive granules and bioerodible ocular minitablets (6 mg and Psi 2 mm). The purpose of this study was to compare minitablets prepared using roller compacted granules with an optimised minitablet formulation, manufactured on laboratory scale by direct compression. The formulation consisted of drum dried waxy maize starch, Carbopol 974P, and ciprofloxacin in a ratio of 90.5/5/3 (w/w/w). Three roller compactor parameters were varied, i.e. the roller speed, the horizontal screw speed and the compaction force, while the vertical screw speed was kept constant. Afterwards, the ribbons were milled to obtain granules suitable for compression. The friability, the flow properties, the bulk material characteristics (apparent and tap density and porosity) and the particle size distributions of two granule sieve fractions (90-125 and 125-355 microm) were investigated. The roller speed and the compaction force have the largest influence on the granule characteristics, followed by the horizontal screw speed. The physical properties of non- and gamma-irradiated minitablets were determined. From the tablet strength, friability and dissolution results, a low compaction force and a high roller speed were shown to be preferable to prepare granules which can be further tabletted into adequate ocular minitablets. PMID- 15760736 TI - Difference in absorption of the two structurally similar flavonoid glycosides, hyperoside and isoquercitrin, in rats. AB - The present study was to investigate oral absorption of the two similar flavonoid glycosides, isoquercitrin (IQ, quercetin-3-O-glucoside) and hyperoside (HP, quercetin-3-O-galactoside) in rats. Two groups of male SD rats received an oral dose of either IQ (4.5 mg/kg) or HP (6.0 mg/kg). Blood samples were collected via jugular vein at time intervals after drug administration and the plasma concentrations of the studied compounds were analyzed by HPLC. The stability of IQ and HP in the GI tract was also measured by incubation with various GI contents from rats. The results showed that unchanged IQ was barely detectable whereas the glucuronidated quercetin (the aglycone of IQ) was found to be the major form in plasma after oral administration of IQ. In contrast, HP could not be detected in plasma neither as unchanged form nor its aglycone or conjugated aglycone form. Additional in vitro stability studies demonstrated that HP is more stable than IQ in the GI tract. This suggests that IQ could be hydrolyzed easier than HP to its aglycone in GI tract before being absorbed. In conclusion, IQ, as a flavonoid glucoside, could be rapidly absorbed and transformed into glucuronidated quercetin and such absorption might be related to the hydrolysis of the type of sugar moieties attached to its aglycone molecule. PMID- 15760737 TI - The influence of gellan gum on the transfer of fluorescein dextran across rat nasal epithelium in vivo. AB - The nasal uptake of a 3000 Da fluorescein dextran (FD3) was investigated in rats, using fluorescence microscopy. The uptake from a formulation containing deacetylated gellan gum, an in situ gelling agent, was compared to that from a mannitol solution. Additionally, the rheological behavior of the gellan gum in water and saline was studied. It was shown that the gellan gum solution was easily administered owing to its low viscosity, and upon contact with the mucosa, a gel was formed. The epithelial uptake and transfer of FD3 appeared to be increased and prolonged using the gellan gum formulation. This increase was not accompanied by qualitative changes of the epithelial FD3 distribution or any visible harmful effects. PMID- 15760738 TI - Effects of process variables on the powder yield of spray-dried trehalose on a laboratory spray-dryer. AB - A systematic examination is presented of the effects of process variables on the powder yield of amorphous trehalose obtained from the Buchi Model 191 laboratory scale mini spray dryer. By using a specially made, narrow cyclone the powder yield could be greatly improved at all process temperatures examined. Calculations of the separation efficiencies of the improved cyclone and the manufacturer's standard cyclone are given, which show that the former's higher tangential particle velocity at the radius of the exit duct is responsible for the improved performance. The powder yield increases with higher process temperatures, owing to improved droplet drying and reduced droplet/particle deposition on the walls of the drying chamber. A maximum in the powder yield is reached, however, after which it decreases sharply. This is caused by heating of the cyclone wall to >10 degrees C above the so-called 'sticky point' of the trehalose, causing increased particle deposits on the walls of the tower and cyclone. Increasing liquid feed flow rate or decreasing atomizing air flow rate too extensively were both detrimental to powder yield. The drying air flow rate should be as high as possible to ensure sufficient enthalpy throughput to dry the trehalose adequately to give a high powder yield. The enthalpy balance calculation for drying trehalose with the new cyclone was used successfully to interpret the results obtained. Some recommendations for optimizing powder yield of an amorphous material are given. PMID- 15760739 TI - Energy analysis of human stumbling: the limitations of recovery. AB - This study has analyzed the segmental energy changes in the recovery from a stumble induced during walking on a treadmill. Three strategies emerged according to the behavior of the perturbed limb, elevating, lowering, and delayed lowering. These three strategies showed different changes in the segmental energy with respect to normal gait. In the elevating strategy, the energy loss induced by the stumble was restored during the perturbed step and reached normal levels during the recovery step. The largest energy changes occurred in the lowering and delayed lowering strategies during the double stance (DS) phase. Moreover, in some of these trials there was energy absorption during the double stance phase for several strides after the perturbation. The most challenging perturbations had longer duration or occurred at mid-swing, triggering delayed lowering or lowering strategies, because more strides and larger energy changes were needed to recover, suggesting a trade-off between stability and energy efficiency. PMID- 15760740 TI - Centre of pressure displacements in trans-femoral amputees during gait. AB - The aim of this study was to describe and quantify centre of pressure (CoP) displacement during walking in 12 unilateral trans-femoral amputees who had worn a prosthesis for at least 10 years. All subjects wore the same type of prosthesis and seven healthy subjects acted as controls. The CoP was acquired by an F-scan system and the displacements along the longitudinal axis of the foot versus time were quantified. An asymmetry of both temporal and spatial parameters of CoP patterns occurred between the sound and the prosthetic foot. The double support time and the time during which the CoP remained in the heel and mid-foot region were longer for the prosthetic limb. Conversely, the stance phase was longer for the sound limb, as was the time spent by the CoP in the forefoot. There was a redistribution of the time during which the CoP remained within the different zones of the sound foot with respect to the normal feet of the control subjects. The asymmetry seen in trans-femoral amputee stance is due not only to an abnormal spatio-temporal distribution of the CoP under the prosthetic leg, but also to a modification of the spatio-temporal distribution of the CoP under the sound foot compared to the normal foot. We conclude that an adaptation occurs in the control of the stance phase on the sound side during amputee gait. PMID- 15760741 TI - Lower extremity kinematics and kinetics during level walking and stair climbing in subjects with triple arthrodesis or subtalar fusion. AB - The purpose of this study was to identify the kinematic and kinetic strategies used by patients with unilateral triple arthrodesis or subtalar fusion during level walking, stair ascent, stair descent and to determine the influence of these different conditions on kinematics and kinetics. Nine subjects with unilateral triple or subtalar fusion and five normal control subjects were recruited for this experiment. Temporal distance, kinematic and kinetic data were collected using a six camera 3-D motion analysis system and a custom fabricated set of stairs with five steps; the second and third steps were each instrumented with one force platform. During level walking, affected limbs lost all of the plantarflexion at the ankle joint during push-off and showed greater knee flexion angle during the same period of stance. During stair ascent, affected limbs showed a different movement pattern at the knee, a greater knee flexion angle during the whole stance phase and a near zero degree of plantarflexion angle during the forward continuance (FCN) phase. During descent, affected limbs showed a greater knee flexion angle during the whole stance phase and less ankle dorsiflexion angle during the same period of stance phase. At the ankle, peak moment and power values were significantly different between the affected side and the limbs of the control subjects during level walking in the push-off phase, stair ascent in the FCN phase, and stair descent in the weight acceptance (WA) phase, where the affected limbs had a lower plantarflexion moment and power values. PMID- 15760742 TI - Change in the locus of dynamic loading axis on the knee joint after high tibial osteotomy. AB - The purpose of this study was to visualise the locus of the dynamic loading axis on the knee joint, and to evaluate changes in this locus during gait after high tibial osteotomy (HTO) in three patients who underwent HTO for medial compartment osteoarthritis (OA) of a varus knee. The bone structure of the lower limb and the relative position of skin markers were acquired from CT images. Motion capture data was acquired using spherical skin markers. Skeletal model movement during gait was calculated based on the movement of the markers. The locus of the dynamic loading axis on the knee joint was defined as the point on the proximal tibia joint surface that intersected with the loading axis of the lower limb, which passed through the centre of the femoral head and the centroid of multiple points surrounded by the distal tibia joint surface contour. This system was able to visualise the locus of the dynamic loading axis on the knee joint and not only lateral but also anterior-posterior direction movement. After HTO, the locus shifted from a medial and posterior area of the medial joint edge of the knee to a central area of the knee joint surface. This indicates that HTO shifted the dynamic loading axis. Lateral movement of the dynamic loading axis in the early stance phase of gait was reduced within a year after HTO. PMID- 15760743 TI - Age and walking speed effects on muscle recruitment in gait termination. AB - During gait termination at normal walking speed, older adults more frequently employ two-step responses, increasing their stopping distance and stopping time more than younger controls. This study investigated ageing effects on lower limb muscle recruitment patterns during stopping at three walking speeds. Twelve young male (26 +/- 3.7 years, range 19-30) and 12 gender-matched older participants (72 +/- 4.3 years, range 65-82) terminated walking at normal, medium and maximum speed. A visual stopping stimulus was presented 10 ms following either left or right heel-contact with no stimulus (catch) on 30% of trials. Electromyographic (EMG) activity was recorded from the tibialis anterior (TA), soleus (SOL), biceps femoris (BF), vastus lateralis (VL) and gluteus medius (GM). Older males more frequently (46% of trials) took two-steps to stop than young males (20%). The stance leg muscles responded significantly faster than the swing leg, and with increased speed, fewer swing limb muscles contributed to stopping. Older males were slower to respond with the stance leg, at 215 ms following the stimulus compared with 176 ms for the younger group. They also recruited fewer swing leg muscles with less frequent activation of the soleus and gluteus medius. Failure to activate muscles would provide less extensor torque to maintain the centre of gravity anterior to the forward base of support. This would decrease the total force opposing horizontal velocity in order to bring the body to rest and, as a consequence, encourage an additional step prior to stopping. PMID- 15760744 TI - Physically active older adults display alterations in gait initiation. AB - An understanding of age-related changes in motor behaviour is important when considering the design of training programs for fall-prevention in the elderly. Gait initiation is a phase of walking during which falls are often provoked and this study compares strategies employed by healthy older and young adults during gait initiation. Twenty-nine older, physically active subjects (65-79 years) and 28 younger individuals (23-40 years) were instructed to cross a staged road, when a traffic light changed from red to green, leading with a freely chosen or predetermined leg. Electromyography of tibialis anterior (TA) and lateral gastrocnemius (LG) muscles, ground reaction forces exerted and step lengths were recorded. The elderly displayed several striking differences compared to the young individuals. Weight bearing during initial standing was considerably more unequal and reaction time was 46% longer, with identical duration of gait initiation. Swing leg peak posterior force (% BW) tended to be smaller, but the increase of vertical force was larger. Eight elderly and one young subject lacked anticipatory TA onset; however TA anticipation increased with trial number in the elderly group. When the starting leg was predetermined, the deficiency in TA anticipation was no longer apparent. In the stance leg all forces were smaller and LG was recruited later, and unlike the young subjects generally after the swing leg had left the ground. The results revealed that ageing leads to significant alterations in weight bearing and ankle muscle activation in relation to gait initiation. Gait initiation changed significantly when the starting leg was predetermined and, in the elderly, with repetition. PMID- 15760745 TI - Initiation of movement from quiet stance: comparison of gait and stepping in elderly subjects of different levels of functional ability. AB - This study describes how elderly subjects initiate gait, and step from a position of quiet stance. Based on scores from selected standardized tests subjects were placed in either a high (HFL) or low functional level (LFL) group and were asked to initiate gait, step onto a 10 cm high, 1.22 m wide curb and step over a 10 cm high, 9 cm wide obstacle at a self paced speed. Stepping conditions affected the velocity of movement. It was clear that all subjects decreased initiation velocity for both curb and obstacle compared to gait initiation. Swing and stance limb acceleration ground reaction forces and EMG amplitude were modulated according to initiation velocity. Toe clearance was greater for obstacle than curb and gait initiation. Swing toe-off was significantly earlier and there was a trend for obstacle clearance to be greater for the HFL group. Those in the LFL group appear to be at a greater risk for falling due to the possible effect of slower rate of toe-off that could influence toe clearance over the obstacle. PMID- 15760746 TI - A comparison of gait with solid and hinged ankle-foot orthoses in children with spastic diplegic cerebral palsy. AB - This study compared the effects of solid and hinged ankle-foot orthoses (AFOs) on the gait of children with spastic diplegic cerebral palsy (CP) who ambulate with excessive ankle plantar flexion during stance. Twelve children with spastic diplegic CP wore no AFOs for an initial 2-week period, solid AFOs for 1 month, no AFOs for 2 weeks, and hinged AFOs for 1 month. Lower extremity muscle timing, knee and ankle joint motions, moments and powers, and temporal-distance characteristics were measured during ambulation for an initial barefoot baseline test, and with solid and hinged AFOs for the other two tests. Both orthoses increased stride length, reduced abnormal ankle plantar flexion during initial contact, midstance and terminal stance (TST), and increased ankle plantar flexor moments closer to normal during TST. Hinged AFOs increased ankle dorsiflexion at TST and increased ankle power generation during preswing (PSW) as compared to solid AFOs, and increased ankle dorsiflexion at loading compared to no AFOs. No other significant differences were found for the gait variables when comparing these orthoses. Either AFO could be used to reduce the excessive ankle plantar flexion without affecting the knee position during stance. The hinged AFO would be recommended to produce more normal dorsiflexion during TST and increased ankle power generation during PSW in children with spastic diplegic CP. PMID- 15760748 TI - Effects of head extension on undisturbed upright stance control in humans. AB - The purpose of the present experiment was to investigate whether and how the head extended posture, commonly encountered in many routine activities, affects undisturbed upright stance control mechanisms in humans. Sixteen young healthy adults stood feet together, with their eyes closed and were asked to sway as little as possible in two Neutral and Extended head conditions. Centre of pressure (CP) displacements, recorded using a force platform, were used to compute the motions of the vertical projection of the centre of gravity (CG(v)) and those of the difference CP - CG(v). A time-domain analysis shows increased mean velocity and surface covered by the trajectory of both elementary motions in the Extended head condition. A frequency analysis also reveals increased root mean squares on the CP - CG(v) motions, suggesting increased muscular activity in the Extended head condition. Furthermore, similar changes occur on CG(v) motions. Finally, modelling these trajectories as a fractional Brownian motion process demonstrates increased spatial transition point co-ordinates at which the corrective process is initiated and a more deterministic control mechanism in this corrective process involving CG(v) motions in the Extended head position. Together, the present findings suggest that head extension position yields a reorganisation of the control mechanisms for maintaining undisturbed upright stance. PMID- 15760747 TI - The heel-contact gait pattern of habitual toe walkers. AB - We used kinematic, kinetic and EMG analysis to compare the spontaneous heel contact gait patterns of 13 children classified as habitual toe walkers (HTWs) and age-matched controls. In the HTWs, the incidence of spontaneous heel-contact strides during a single recording session ranged from 15% to 92%, with no correlation with age, passive ankle joint excursion, walking speed and trial order. Hallmarks of the heel-contact strides were premature heel-rise, reversal of the second rocker, relative shortening of the loading response and anticipation and enhancement of the electromyographic (EMG) activity normally observed in the triceps surae (TS) during the first half of the stance phase. This variant of the locomotor program is different from the walking patterns observed in normally developing toddlers and children with cerebral palsy (CP). It does not necessarily reflect a functional adaptation to changes in the rheological properties of the muscle-tendon complex. PMID- 15760749 TI - Errors in measuring sagittal arch kinematics of the human foot with digital fluoroscopy. AB - Although fluoroscopy has been used to evaluate motion of the foot during gait, the accuracy and precision of fluoroscopic measures of osseous structures of the foot has not been reported in the literature. This study reports on a series of experiments that quantify the magnitude and sources of error involved in digital fluoroscopic measurements of the medial longitudinal arch. The findings indicate that with a global distortion correction procedure, errors arising from image distortion can be reduced threefold to 0.2 degrees for angular measurements and to 0.1 mm for linear measures. The limits of agreement for repeated angular measures of the calcaneus and first metatarsal were +/-0.5 degrees and +/-0.6 degrees , indicating that measurement error was primarily associated with the manual process of digitisation. While the magnitude of the residual error constitutes about +/-2.5% of the expected 20 degrees of movement of the calcaneus and first metatarsal, out-of-plane rotation may potentially contribute the greatest source of error in fluoroscopic measures of the foot. However, even at the extremes of angular displacement (15 degrees ) reported for the calcaneum during running gait, the root mean square (RMS) error was only about 1 degrees . Thus, errors associated with fluoroscopic imaging of the foot appear to be negligible when compared to those arising from skin movement artefact, which typically range between 1.5 and 4 mm (equating to errors of 2 degrees to 17 degrees for angular measures). Fluoroscopy, therefore, may be a useful technique for analysing the sagittal movement of the medial longitudinal arch during the contact phase of walking. PMID- 15760750 TI - Relationship between energy cost, gait speed, vertical displacement of centre of body mass and efficiency of pendulum-like mechanism in unilateral amputee gait. AB - The energy cost of gait (C) is greater in amputee than in normal subjects. Our objective was to assess the influence of lower speed, inefficient pendulum mechanism and disturbed smoothness of centre of body mass (CM(b)) displacement on C in unilateral amputees and to have a better understanding the relationship between these variables. Twelve adult patients (six traumatic transfemoral and six vascular transtibial amputees) participated in the study. Lower limb kinematics, displacement of CM(b), mechanical work done by the muscles to move the CM(b) and the segments due to their movements relative to the CM(b), efficiency of the pendulum mechanism, and C were assessed simultaneously in the 12 amputees walking at their self-selected speed. Our results show that C depended on gait speed, and efficiency of pendulum-like mechanism of walking but did not depend on the smoothness of CM(b). The use of only a single variable to explain the extra cost in amputee gait could sometimes be misleading. PMID- 15760751 TI - Soft tissue artefact assessment in humeral axial rotation. AB - The accuracy of upper-limb kinematic data acquired from optoelectronic systems with retro-reflective markers is poor, mainly due to soft tissue artefact (STA). For the upper-arm, humeral internal/external rotation (HIER) is the movement most affected by STA, which is measured as a percentile fraction (K) of the effective humeral axial rotation performed. The aim of this work was to quantify STA during HIERs, with independently varying attitude of the humerus and elbow flexion, and to test the possibility of estimating its mean value over the tested upper-limb orientations using one simple trial. Six able-bodied subjects performed a series of HIERs in combination with elbow flexion for different humeral planes and degrees of elevation. During the trials the instantaneous attitudes of two humeral anatomical frames were compared, one being affected by the STA to be measured, and the other assumed as the gold standard. K was found to range from 20% to 48% of the effective humeral axial rotation performed, depending on the subject, humeral attitude and elbow flexion. These last two factors comparably affect STA and resulted in mean K coefficients of variation among the subjects of about 9% and 7%, respectively. Common patterns of K with elbow flexion and humerus elevation are discussed. The data also show that the mean of K of a subject is very close to the value assessed in a specific upper-limb configuration consistent among the subjects. This result from this study could be used to build up a time-saving STA compensation procedure suitable for clinical applications. PMID- 15760752 TI - Froude and the contribution of naval architecture to our understanding of bipedal locomotion. AB - It is fascinating to think that the ideas of two 19th century naval architects could offer useful insights for 21st century scientists contemplating the exploration of our planetary system or monitoring the long-term effects of a neurosurgical procedure on gait. The Froude number, defined as Fr = v2/gL, where v is velocity, g is gravitational acceleration and L is a characteristic linear dimension (such as leg length), has found widespread application in the biomechanics of bipedal locomotion. This review of two parameters, Fr and dimensionless velocity beta = (Fr)1/2, that have served as the criterion for dynamic similarity, has been arranged in two parts: (I) historical development, including the contributions by William Froude and his son Edmund, two ship designers who lived more than 130 years ago, the classic insights of D'Arcy Wentworth Thompson who, in his magnum opus On Growth and Form, espoused the connection between mathematics and biology, and the pioneering efforts of Robert McNeill Alexander, who popularised the application of Fr to animal locomotion; and (II) selected applications, including a comparison of walking for people of different heights, exploring the effects of different gravitational fields on human locomotion, establishing the impact of pathology and the benefits of treatment, and understanding the walking patterns of bipedal robots. Although not all applications of Fr to locomotion have been covered, the review offers an important historical context for all researchers of bipedal gait, and extends the idea of dimensionless scaling of gait parameters. PMID- 15760754 TI - MR angiography physics: an update. PMID- 15760755 TI - MR angiography interpretation: techniques and pitfalls. PMID- 15760756 TI - MR angiography of the thoracic aorta. PMID- 15760757 TI - MR angiography of the abdominal aorta. PMID- 15760758 TI - MR angiography of the peripheral arteries. PMID- 15760759 TI - MR venography. PMID- 15760760 TI - MR angiography in patients with renal disease. PMID- 15760761 TI - Role of MR angiography in vascular interventional planning. PMID- 15760762 TI - Contrast-enhanced MR angiography in infants and children. PMID- 15760764 TI - Emerging functional MR angiographic techniques. PMID- 15760763 TI - Atherosclerosis imaging using MR imaging: current and emerging applications. PMID- 15760765 TI - Contrast agents: innovations and potential applications for body MR angiography. PMID- 15760766 TI - Gene dosage and pathogenesis of Parkinson's disease. AB - Four recent papers related specifically to the familial form of Parkinson's disease reinforce the idea that endogenous levels of alpha-synuclein can strongly influence disease phenotype. Two recent publications of alpha-synuclein duplication mutations show that the severity of familial Parkinsonian phenotype is dependent upon SNCA gene dosage and corresponding protein levels. Familial point mutations in SNCA were found to impair the efficient lysosomal degradation of alpha-synuclein, potentially resulting in elevated levels of alpha-synuclein. Conversely, the complete knockout of SNCA has little effect on transgenic mice. It is now clear that the regulation of alpha-synuclein levels has potential significance in the pathogenesis and treatment of sporadic PD. PMID- 15760767 TI - TIMP-2: an endogenous inhibitor of angiogenesis. AB - Remodeling of the extracellular matrix--regulated by the matrix metalloproteinases (MMPs) and their endogenous inhibitors--is an important component of disease progression in many chronic disease states. Unchecked MMP activity can result in significant tissue damage, facilitate disease progression and is associated with host responses to pathologic injury, such as angiogenesis. The tissue inhibitors of metalloproteinases (TIMPs) have been shown to regulate MMP activity. However, recent findings demonstrate that an MMP-independent effect of TIMP-2 inhibits the mitogenic response of human microvascular endothelial cells to growth factors. This is the first demonstration of a cell-surface signaling receptor for a member of the TIMP family and suggests that TIMP-2 functions to regulate cellular responses to growth factors. These new findings are integrated in a comprehensive model of TIMP-2 function in tissue homeostasis. PMID- 15760768 TI - Modulation of angiogenesis with siRNA inhibitors for novel therapeutics. AB - Cancer and many other serious diseases are characterized by the uncontrolled growth of new blood vessels. Recently, RNA interference (RNAi) has reinvigorated the therapeutic prospects for inhibiting gene expression and promises many advantages over binding inhibitors, including high specificity, which is essential for targeted therapeutics. This article describes the latest developments using small-interfering RNA (siRNA) inhibitors to downregulate various angiogenic and tumor-associated factors, both in cell-culture assays and in animal disease models. The majority of research efforts are currently focused on understanding gene function, as well as proof-of-concept for siRNA-mediated anti-angiogenesis. The prospects for siRNA therapeutics, both advantages and looming hurdles, are evaluated. PMID- 15760769 TI - Chemokines and Toll-like receptors in Lyme disease pathogenesis. AB - Lyme disease is a tick-transmitted inflammatory disorder, caused by the spirochete Borrelia burgdorferi (Bb). Recent discoveries cast new light on Bb dissemination and the ensuing pathogenesis of inflammation. Although the strong proinflammatory Bb lipoproteins have been implicated in the induction of inflammation, they do not seem to act exclusively through Toll-like receptor (TLR) engagement. In fact, mice that are deficient for MyD88, a component of the TLR signaling pathway, manifest similar or increased recruitment of cells into Bb infected tissues. By contrast, the absence of the chemokine receptor CXCR2 results in reduced inflammation. Overall, these findings highlight the complexity of Lyme disease pathogenesis and identify chemokine pathways as novel therapeutic targets for the control of Bb-induced inflammation. PMID- 15760771 TI - Genetic disorders of the LRP5-Wnt signalling pathway affecting the skeleton. AB - Osteoporosis is a common, increasingly prevalent and potentially debilitating condition of men and women. Genetic factors are major determinants of bone mass and the risk of fracture, but few genes have been definitively demonstrated to be involved. The identification of these factors will provide novel insights into the processes of bone formation and loss and thus the pathogenesis of osteoporosis, enabling the rational development of novel therapies. In this article, we present the extensive genetic and functional data indicating that the LRP5 gene and the Wnt signalling pathway are key players in bone formation and the risk of osteoporosis, and that LRP5 signalling is essential for normal morphology, developmental processes and bone health. PMID- 15760770 TI - Protein kinase A as a therapeutic target for memory disorders: rationale and challenges. AB - cAMP-dependent protein kinase A (PKA) signaling has a key role in memory processes and has been identified as a potential therapeutic target for memory disorders. The activation of PKA signaling is crucial for the consolidation of long-term memories dependent on the hippocampus and/or the amygdala, By contrast, initial studies indicate that cAMP-PKA activation might impair the working memory and executive functions of the prefrontal cortex. Furthermore, PKA activation in the nucleus accumbens might increase sensitivity to addiction. These complexities must be heeded when designing medications aimed at altering PKA activity. PKA might be most practical as a therapeutic target in disorders with global alterations in cAMP-PKA activity due to genetic or environmental factors. PMID- 15760772 TI - Dissecting the genetic etiology of major depressive disorder using linkage analysis. AB - Major depressive disorder (MDD) is clinically and genetically heterogeneous. Studies suggest that recurrence, early onset and comorbid phenotypes define more genetically homogeneous sub-samples. The concordance of linkage findings in recent studies using such approaches is encouraging. Sex-specific analyses and broader phenotypes have also yielded interesting results. These findings indicate that future research should consider comorbid disorders and sex-specific analyses. However, this direction must be approached with caution, owing to the complex multiple-testing issues that arise when considering numerous related phenotypes. With appropriate interpretation, these findings indicate a new potential for positional cloning efforts to locate genes in consensus regions. Genes found might influence specific subtypes of MDD or broader phenotypes, leading to enhanced clinical characterization and management of MDD. PMID- 15760773 TI - Assay of free thyroxine and free triiodothyronine in fine-needle aspiration of thyroid nodules: a useful and low-cost assessment. AB - OBJECTIVE: To evaluate whether analysis of thyroid hormones in fine-needle aspiration (FNA) of thyroid nodules can provide information about the functional status and the nature of the nodules. METHODS: We studied 4 groups of patients: group 1, 17 patients with autonomous hyperfunctioning thyroid nodules; group 2, 52 patients with cold nonfunctioning thyroid nodules; group 3, 12 patients with malignant thyroid nodules; and group 4 (control group), 10 patients with nonthyroid nodular lesions (enlarged parathyroid glands or lymph nodes). The assay of thyroid hormones was performed in FNA after the washing of needles and, with patient consent, also in normal thyroid parenchyma. RESULTS: The free thyroxine (FT(4)) and free triiodothyronine (FT(3)) values were remarkably high in group 1 (mean, 5.5 +/- 0.53 ng/dL and 27.6 +/- 3.1 pg/mL, respectively; P<0.05 versus group 2 and group 4, the control group). The levels of FT(4) and FT(3) were very low in group 3 (<0.2 ng/dL and <1.0 pg/mL, respectively; P<0.05 versus group 2). Thyroglobulin values in FNA specimens were much higher than the normal range in human serum, but no significant differences were found between the various groups. The control group had low levels of FT(4) and FT(3) (<0.2 ng/dL and <1.0 pg/mL, respectively) in conjunction with low levels of thyroglobulin, whereas parathyroid hormone levels were high in parathyroid nodules. CONCLUSION: These results show that assay of FT(4) and FT(3) in FNA can yield information about the functional status of thyroid nodules and, indirectly, about the nature of nodules. In this era of sophisticated new molecular markers in FNA cytology, this low-cost diagnostic method can be readily performed in every laboratory. PMID- 15760774 TI - Management of nondiagnostic thyroid fine-needle aspiration biopsy: survey of endocrinologists. AB - OBJECTIVE: To evaluate the approach of endocrinologists in the setting of nondiagnostic thyroid fine-needle aspiration (FNA) biopsies. METHODS: In 2002, we surveyed physicians attending the national annual meetings of the American Association of Clinical Endocrinologists and the Endocrine Society of North America, using a 13-item questionnaire. The responses were tallied and analyzed. RESULTS: Of the 143 respondents, 139 were endocrinologists, with a male:female ratio of 2.5:1. Most respondents were involved in a medical practice in North America, but Europe, Asia, New Zealand, and Australia were also represented. Of those performing thyroid FNA biopsy, 31% used thyroid ultrasound guidance. Among the survey respondents, 16%, 49%, 20%, and 15% performed less than 2, 2 to 5, 6 to 10, and more than 10 thyroid FNA biopsies per month, respectively. Among the respondents, 13.5%, 44%, 28.5%, 10%, and 4% had non-diagnostic rates of less than 5%, 5 to 10%, 11 to 20%, 21 to 30%, and more than 30%, respectively. The approach of the respondents to an initially nondiagnostic FNA was repeated FNA biopsy in 87%, observation in 7%, levothyroxine suppression in 4%, and thyroid scintigraphy in 2%. Respondents believed that the most cost-effective approach in a patient with nondiagnostic FNA was repeated biopsy (82%), monitoring the size of the thyroid nodule (17%), and surgical referral (<1%). No one was willing to repeat the thyroid biopsy more than three times. CONCLUSION: On the basis of findings in our survey, most endocrinologists repeat thyroid FNA at least once when confronted with a nondiagnostic result. No published studies have demonstrated the cost-effectiveness of this approach versus proceeding to surgical intervention or observation. We hope that this survey will encourage further studies on this issue. PMID- 15760775 TI - What is hypoglycemia in patients with well-controlled type 1 diabetes treated by subcutaneous insulin pump with use of the continuous glucose monitoring system? AB - OBJECTIVE: To analyze 72-hour interstitial glucose concentrations measured by a glucose sensor in adult subjects. METHODS: We compared glucose levels in 10 patients with type 1 diabetes mellitus (DM-1) and 10 normal control subjects with use of the Medtronic Continuous Glucose Monitoring System (CGMS). Hypoglycemic events identified by the CGMS, the patients, or both were compared in the patients with DM-1. In addition, the results between two glucose sensors placed at similar sites in the same subject were evaluated. RESULTS: Hemoglobin A1c and mean glucose values were higher in the DM-1 group than in the control group (P<0.01), whereas time spent at glucose levels of less than 60, 60 to 79, and 121 to 150 mg/dL were similar between the two study groups. Time spent at glucose levels above 150 mg/dL was greater in the DM-1 group than in the control group (P<0.05). Of the 74 total hypoglycemic events in the patients with DM-1, the sensor and the patient recognized 14%, only the patient recognized 9% (confirmed by readings of <55 mg/dL on a glucose meter), and only the sensor detected 77%. Of note, 41% of the patient-identified symptomatic hypoglycemic events were missed by the CGMS. When one patient with DM-1 and one control subject wore two glucose sensors simultaneously, one and not the other sensor often identified "hypoglycemia," and the correlation was only fair to good by intraclass correlation analysis. CONCLUSION: Many asymptomatic "hypoglycemic events" identified by the CGMS may not be actual hypoglycemia. Hypoglycemia is both overreported and underreported with use of the CGMS. Thus, adjustment of insulin doses to reduce asymptomatic hypoglycemia identified by the CGMS alone may not be warranted. PMID- 15760776 TI - Thyroid cytology and risk of thyroid cancer: differences among indeterminate specimens. AB - OBJECTIVE: To assess the potential for stratification of indeterminate cytologic findings on fine-needle aspiration (FNA) of thyroid nodules in an effort to improve therapeutic strategies. METHODS: We attempted to determine the malignant risk associated with various indeterminate FNA cytologic patterns by correlation of specimens with the final histologic diagnosis. For this analysis, we identified 294 computerized medical records of surgically treated thyroid nodules during a 5-year period at our institution with the corresponding FNA cytology reports available. RESULTS: Of the 294 surgical cases, 162 with a positive or indeterminate cytologic report were selected, reviewed, and classified. Of 52 patients with positive cytologic findings on FNA, 51 (98%) had a final histologic report of a malignant thyroid nodule. Of 110 patients with indeterminate specimens, 30 (27%) had a final histologic diagnosis of thyroid carcinoma. The presence of nuclear atypia was predictive of thyroid carcinoma in 75% of patients, a Hurthle cell cytologic pattern was associated with a malignant thyroid nodule in 33%, and a hypercellular smear was suggestive of malignant involvement in 26% of cases. The lowest rate of malignant potential was associated with cytologic microfollicular and scant colloid alone subtype (6%). CONCLUSION: The results of this study show that indeterminate thyroid cytologic specimens can be subdivided into groups with different malignant risks. A microfollicular cytologic pattern in the absence of a hypercellular smear or nuclear atypia does not support a recommendation of surgical treatment. A malignant cytologic diagnosis has a high positive predictive value for detection of thyroid cancer. PMID- 15760777 TI - Osteogenic sarcoma of the sella after radiation treatment of a pituitary adenoma. AB - OBJECTIVE: To report an uncommon case of osteogenic sarcoma of the sella turcica after radiation treatment of a pituitary adenoma. METHODS: We present the clinical history, physical findings, laboratory data, imaging studies, and pathologic findings in a patient found to have osteogenic sarcoma of the sella after radiation therapy for a nonfunctioning pituitary adenoma. RESULTS: Six years after transsphenoidal resection and postoperative fractionated radiation therapy for a nonfunctioning pituitary adenoma that extended to the cavernous sinus, a 45-year-old man presented with a sinus infection, diplopia, and ophthalmoplegia of the right eye. A computed tomographic scan of the head showed a mass in the sella with involvement of the optic chiasm and right cavernous sinus. Transsphenoidal resection and debulking of the tumor revealed an osteogenic sarcoma. The patient was discharged from the hospital with residual diplopia and ophthalmoplegia. He was treated with levothyroxine, testosterone, and hydrocortisone. Six weeks later, the patient was readmitted after he was found unresponsive, and computed tomographic scans disclosed a massive cerebrovascular accident. He died a few days later. CONCLUSION: Osteogenic sarcoma is a rare, late complication of radiation treatment of pituitary adenoma. Although radiotherapy remains an effective adjunctive treatment in patients with pituitary adenomas, particularly those with residual or recurrent tumor, potential complications must be acknowledged. PMID- 15760778 TI - Reversible renal insufficiency attributable to thyroid hormone withdrawal in a patient with type 2 diabetes mellitus. AB - OBJECTIVE: To present a case of papillary thyroid carcinoma and the need for follow-up surveillance in a patient with type 2 diabetes mellitus who was being treated with metformin. METHODS: The clinical and laboratory findings are summarized, and results of whole-body scanning are described. A discussion addresses the importance of monitoring renal function during thyroid hormone withdrawal for scintigraphic imaging with use of radioactive iodine. RESULTS: A 60-year-old man with type 2 diabetes was diagnosed with metastatic papillary thyroid carcinoma, and renal insufficiency developed during withdrawal from thyroid hormone for surveillance 131 I whole-body scanning. An increase in thyroglobulin attributable to recombinant human thyrotropin stimulation led to detection of persistent cancer, and thyroid hormone withdrawal was begun in preparation for further 131 I treatment. Pretherapy laboratory results revealed a thyrotropin level of 46.1 mIU/mL, a thyroglobulin level of 13.7 ng/mL, and an increased serum creatinine (Cr) concentration from a baseline of 0.9 mg/dL to 1.9 mg/dL while the creatinine clearance (CrCl) was 67 mL/min. Therapy with metformin was immediately discontinued. On the basis of results from 131 I whole-body dosimetry, the patient was treated with 250 mCi of 131 I. Levothyroxine therapy was resumed approximately 72 hours after 131 I therapy, and blood tests 2 weeks thereafter revealed a Cr of 1.1 mg/dL and a CrCl of 105 mL/min. The metformin treatment was then reinitiated without any complications. CONCLUSION: Patients with hypothyroidism can have up to a 20 to 30% decrease in glomerular filtration rate, which tends to resolve when euthyroidism is achieved. In patients with diabetes taking metformin who have hypothyroidism for any reason including thyroid hormone withdrawal, serum Cr levels should be monitored until thyroid hormone levels normalize. Metformin therapy should be discontinued in patients with significant renal impairment, defined as either a serum Cr level of 1.5 mg/dL or more in men and 1.4 mg/dL or more in women or a CrCl of less than 60 mL/min. PMID- 15760779 TI - Multiple myeloma presenting with recurrent hypercalcemia in a patient with a history of primary hyperparathyroidism: report of case and review of literature. AB - OBJECTIVE: To report a case of primary hyperparathyroidism (PHPT) that presented with recurrent hypercalcemia due to multiple myeloma after successful parathyroidectomy. METHODS: The initial manifestations, investigations, and postoperative follow-up of a case of hypercalcemia due to PHPT are described. The studies performed for evaluation for recurrent hypercalcemia and the subsequent diagnosis of multiple myeloma are discussed. The association between these disorders and reports of similar cases in the literature are reviewed. RESULTS: A 72-year-old woman was referred for incidentally discovered hypercalcemia. She had no history of kidney stones or fractures. Further investigations revealed a high parathyroid hormone (PTH) level, hypercalciuria, and low bone mass, particularly at the cortical sites. Parathyroidectomy was performed, and a right inferior parathyroid adenoma was removed. Postoperatively, both the calcium and PTH levels normalized. She presented 9 months later with a 3-week history of pain in her left hip, polyuria, nausea, and vomiting. The patient had severe hypercalcemia and a suppressed PTH level. Further investigations revealed multiple bony lytic lesions, abnormalities on serum protein electrophoresis, and features consistent with multiple myeloma on a bone marrow biopsy specimen. CONCLUSION: Hypercalcemia can occur in patients with PHPT and multiple myeloma; however, the occurrence of both disorders in the same patient is rare. Review of the literature revealed only a few cases similar to ours. Evidence in the literature suggests that monoclonal gammopathies occur more often in patients with PHPT than in the general population; therefore, screening for monoclonal gammopathy may be warranted in patients with PHPT. PMID- 15760780 TI - Some clues and pitfalls in the diagnosis of acromegaly. AB - OBJECTIVE: To describe a case of acromegaly in a young woman using oral contraceptives who had nonspecific symptoms and persistent hyperphosphatemia and to analyze the reasons for delay in diagnosis. METHODS: A 31-year-old woman underwent evaluation for failure of menstrual flow to resume after discontinued use of oral contraceptives. Clinical and laboratory findings are summarized, and atypical manifestations of acromegaly are discussed. RESULTS: The patient complained of weight gain, acne, and increased facial hair growth, and laboratory tests showed a low estradiol level and hyperinsulinemia. "Post-pill amenorrhea" and polycystic ovary syndrome were considered possible diagnoses. During subsequent endocrinology consultation, hyperphosphatemia was present on three occasions during a 7-month period. Hormonal studies for evaluation of pituitary function revealed increased insulin-like growth factor-I and growth hormone (GH) levels. Acromegaly was suspected, and the increased GH level failed to suppress after a glucose load. Magnetic resonance imaging revealed a pituitary macro adenoma with suprasellar extension. Visual field testing showed bitemporal abnormalities. Transsphenoidal resection successfully removed the GH-producing tumor, and hormone replacement therapy was initiated postoperatively. CONCLUSION: The textbook presentation of GH excess is a late feature of acromegaly. The diagnosis in a young woman can be easily missed when clinicopathologic findings are not classic and the picture is further clouded by factors such as oral contraceptive use. Features may also resemble polycystic ovary syndrome, a more common condition in women of childbearing age. This case also illustrates that hyperphosphatemia, overlooked in this patient, can precede the full-blown clinical manifestations of acromegaly and serve as a subtle clue to the underlying disease. Awareness of these situations and screening for high GH levels in patients with unexplained, persistent hyperphosphatemia are advisable for making an early correct diagnosis and providing appropriate therapy. PMID- 15760781 TI - Role of insulin-like growth factor-I in diagnosis and management of acromegaly. AB - OBJECTIVE: To highlight the use of insulin-like growth factor-I (IGF-I) measurements in the management of acromegaly. METHODS: The physiologic and pathophysiologic regulation of growth hormone (GH) and IGF-I secretion and the role of measurements of these two variables for diagnosing and monitoring the acromegalic condition are discussed. RESULTS: Chronic GH hypersecretion by pituitary tumors leads to the clinical condition of acromegaly, characterized by enlargement of membranous bones and soft tissue overgrowth. Because of the frequent delay in diagnosis of acromegaly, disease management often involves the use of multiple therapeutic modalities. Therefore, adequate patient management necessitates precise diagnostic tests to (1) establish that the acromegalic condition exists, (2) determine the degree of severity, and (3) assess the effectiveness of treatment in patients who have received therapy. Because the predicted mortality rate among patients who have achieved a biochemical remission is substantially improved in comparison with those who have biochemical indices that indicate residual disease activity after therapy, a biochemical test is required to determine the necessity of undertaking further treatment. The GH dependent peptide IGF-I provides useful information for making this decision. In addition to providing data about the degree of GH hypersecretion, IGF-I concentrations reflect the severity of disease at the time of initial assessment and the degree of improvement in response to any therapeutic intervention. Direct measurement of GH after glucose suppression is the best indicator of residual tumor secretory activity. IGF-I measurements should be performed in a reference laboratory with adequate age-adjusted normative ranges and a high degree of precision, which allows serial determinations over time to reflect the degree of biochemical improvement. Measurement of IGF-I often correlates with the degree of persistent GH hypersecretory abnormality after therapy, although no precise correlation is found in some cases. CONCLUSION: Measurements of GH suppression after glucose administration and of IGF-I after treatment of patients with acromegaly provide useful complementary information. Every effort should be made to achieve normalization of these biochemical indices so that patients are offered the best therapeutic prognosis. PMID- 15760782 TI - Milk-alkali syndrome from ingestion of calcium carbonate in a patient with hypoparathyroidism. PMID- 15760783 TI - Remembrance of things adrenal: a historical perspective. PMID- 15760784 TI - Number by numbers. PMID- 15760785 TI - Visual vignette. PMID- 15760786 TI - Is subclinical hypothyroidism associated with hyperhomocysteinemia? AB - OBJECTIVE: To evaluate whether subclinical hypothyroidism is associated with hyperhomocysteinemia, in an attempt to determine whether hyperhomocysteinemia may explain the observed increased risk of coronary artery disease in patients with subclinical hypothyroidism, as has been shown in patients with overt hypothyroidism. METHODS: We prospectively enrolled consecutive patients with newly diagnosed subclinical hypothyroidism and a parallel group of euthyroid control subjects in this study. Subclinical hypothyroidism was defined as high thyrotropin and normal free thyroxine levels. Fasting plasma total homocysteine, thyrotropin, and free thyroxine levels were measured in all participants, and persistent subclinical hypothyroidism was confirmed at least once. RESULTS: Forty seven patients (42 women and 5 men; mean age, 38 +/- 15 years) with subclinical hypothyroidism and 50 control subjects (46 women and 4 men; mean age, 34 +/- 10 years) were enrolled in the study. The manufacturer's reference range for the total homocysteine assay was 5 to 15 micromol/L, and we defined our own reference range as 3.9 to 10.8 micromol/L. The mean values (+/-SE) for total homocysteine in patients and control subjects were 7.44 +/- 0.5 micromol/L and 7.22 +/- 0.2 micromol/L, respectively (P = 0.68). CONCLUSION: In this study, we found no association between subclinical hypothyroidism and hyperhomocysteinemia. Other contributing factors, such as associated hyperlipidemia, may explain the observed increased risk of coronary artery disease in patients with subclinical hypothyroidism. PMID- 15760787 TI - Symptoms in patients with primary hyperparathyroidism: muscle weakness or sleepiness. AB - OBJECTIVE: To evaluate the effects of parathyroidectomy on the nonspecific symptoms associated with primary hyperparathyroidism, with an emphasis on distinguishing sleepiness from muscle weakness. METHODS: We conducted a prospective cohort study of consecutive patients presenting for parathyroidectomy for the treatment of primary hyperparathyroidism. Twenty-three patients underwent assessment preoperatively and postoperatively. The Functional Outcomes of Sleep Questionnaire (FOSQ), the Epworth Sleepiness Scale (ESS), and a questionnaire of disease-specific symptoms were administered preoperatively, 2 weeks postoperatively, and 4 to 6 months postoperatively. The FOSQ assesses how sleepiness alters daytime performance, and the ESS reflects the degree of daytime sleepiness. RESULTS: All 23 patients achieved normocalcemia postoperatively. Although no significant changes were found in the overall scores for the FOSQ and ESS after parathyroidectomy, there was a significant decline in the overall mean incidence of disease-specific symptoms postoperatively (P